Discovery and characterization of [(cyclopentyl)ethyl]benzoic acid inhibitors of microsomal prostaglandin E synthase-1.

We describe a novel class of acidic mPGES-1 inhibitors with nanomolar enzymatic and human whole blood (HWB) potency. Rational design in conjunction with structure-based design led initially to the identification of anthranilic acid 5, an mPGES-1 inhibitor with micromolar HWB potency. Structural modifications of 5 improved HWB potency by over 1000×, reduced CYP2C9 single point inhibition, and improved rat clearance, which led to the selection of [(cyclopentyl)ethyl]benzoic acid compound 16 for clinical studies. Compound 16 showed an IC80 of 24nM for inhibition of PGE2 formation in vitro in LPS-stimulated HWB. A single oral dose resulted in plasma concentrations of 16 that exceeded its HWB IC80 in both rat (5mg/kg) and dog (3mg/kg) for over twelve hours.

Characterization of 3,3-dimethyl substituted N-aryl piperidines as potent microsomal prostaglandin E synthase-1 inhibitors.

Here we report on novel, potent 3,3-dimethyl substituted N-aryl piperidine inhibitors of microsomal prostaglandin E synthases-1(mPGES-1). Example 14 potently inhibited PGE2 synthesis in an ex vivo human whole blood (HWB) assay with an IC50 of 7nM. In addition, 14 had no activity in human COX-1 or COX-2 assays at 30µM, and failed to inhibit human mPGES-2 at 62.5µM in a microsomal prep assay. These data are consistent with selective mPGES-1-mediated reduction of PGE2. In dog, 14 had oral bioavailability (74%), clearance (3.62mL/(min*kg)) and volume of distribution (Vd,ss=1.6L/kg) values within our target ranges. For these reasons, 14 was selected for further study.

Effect of Food Consumption on the Pharmacokinetics, Safety, and Tolerability of Once-Daily Orally Administered Orforglipron (LY3502970), a Non-peptide GLP-1 Receptor Agonist.

INTRODUCTION: We assessed the effect of the prandial state on the pharmacokinetics, safety, and tolerability of single and multiple doses of orforglipron (LY3502970), an oral, non-peptide glucagon-like peptide 1 receptor agonist (GLP-1 RA), in two studies (A and B). METHODS: Study A and study B were phase 1, randomized, crossover studies in healthy adults aged 18-65 years and 21-70 years, respectively. Participants received single (3 mg, study A) or multiple (16 mg, study B) oral doses of orforglipron under fasted and fed conditions. Blood samples were collected pre- and postdose to assess area under the concentration-time curve (AUC), maximum observed drug concentration (Cmax), time of Cmax (tmax), and half-life (t1/2) associated with terminal rate constant. AUC and Cmax were analyzed using a linear mixed-effects model. Treatment differences were presented as ratios of geometric least squares means (GLSM). Treatment-emergent adverse events (TEAEs), adverse events of special interest, and serious adverse events were assessed. RESULTS: Study A included 12 participants (mean age 45.0 years; male 66.7%); study B included 34 participants (mean age 42.8 years; male 88.2%). GLSM AUC and Cmax were lower by 23.7% and 23.2% in study A, and 17.6% and 20.9% in study B, in the fed versus fasted states, respectively. In both studies, t1/2 and median tmax were comparable between fed and fasted states. The majority of TEAEs in both studies were gastrointestinal tract-related conditions. No serious adverse events or deaths were reported in either study. CONCLUSION: The observed pharmacokinetic differences due to the prandial state are unlikely to contribute to clinically meaningful differences in the efficacy of orforglipron. The safety profile was consistent with the known profiles of other GLP-1 RAs. Given the absence of prandial restrictions, orforglipron may emerge as a convenient oral treatment option for patients with type 2 diabetes or obesity. TRIAL REGISTRATION: ClinicalTrials.gov identifiers, NCT03929744 and NCT05110794.

Progressive multifocal leukoencephalopathy (PML) associated with HIV Clade C--is not uncommon.

Progressive multifocal leukoencephalopathy (PML) is a rare, subacute, demyelinating disease of the central nervous system caused by JC virus. Studies of PML from HIV Clade C prevalent countries are scarce. We sought to study the clinical, neuroimaging, and pathological features of PML in HIV Clade C patients from India. This is a prospective cum retrospective study, conducted in a tertiary care Neurological referral center in India from Jan 2001 to May 2012. Diagnosis was considered "definite" (confirmed by histopathology or JCV PCR in CSF) or "probable" (confirmed by MRI brain). Fifty-five patients of PML were diagnosed between January 2001 and May 2012. Complete data was available in 38 patients [mean age 39 ± 8.9 years; duration of illness-82.1 ± 74.7 days). PML was prevalent in 2.8 % of the HIV cohort seen in our Institute. Hemiparesis was the commonest symptom (44.7 %), followed by ataxia (36.8 %). Definitive diagnosis was possible in 20 cases. Eighteen remained "probable" wherein MRI revealed multifocal, symmetric lesions, hypointense on T1, and hyperintense on T2/FLAIR. Stereotactic biopsy (n = 11) revealed demyelination, enlarged oligodendrocytes with intranuclear inclusions and astrocytosis. Immunohistochemistry revelaed the presence of JC viral antigen within oligodendroglial nuclei and astrocytic cytoplasm. No differences in clinical, radiological, or pathological features were evident from PML associated with HIV Clade B. Clinical suspicion of PML was entertained in only half of the patients. Hence, a high index of suspicion is essential for diagnosis. There are no significant differences between clinical, radiological, and pathological picture of PML between Indian and Western countries.

A Prospective Multi-centric Study of Acceptance, Insertion and Follow-Up of Postpartum Insertions of IUCD.

Purpose of the Study: The unmet need for contraception in the postpartum period is a major challenge in our country. Unintended pregnancies are highest in the first year after birth, and postpartum IUCD insertion is an effective way to counter this problem. This study was planned to build up data for acceptance and follow-up of postpartum IUCD insertions. Methods: The present study has included data of PPIUCD insertions and follow-up from seven institutions over a period of 6 months. The case recruitment lasted for 3 months, including only those who had PPIUCD insertions in this period, and they were followed up for a period of 6 months. The follow-up of patients was at 6 weeks and 6 months. All issues were addressed including side effects, expulsions, myths surrounding the device, etc., along with routine postnatal care. Results and Conclusion: There were 5227 deliveries and 1895 insertions. The acceptance rate was 36%, and a follow-up at 6 weeks and 6 months showed up an expulsion rate of approximately 4% and a removal rate of 5%. Overall, at the end of 6 months we have a continuation rate of 90%. This shows that a dedicated approach to postpartum contraception will definitely bring down incidence of unintended pregnancies.

Mechanistic and pharmacological characterization of PF-04457845: a highly potent and selective fatty acid amide hydrolase inhibitor that reduces inflammatory and noninflammatory pain.

The endogenous cannabinoid (endocannabinoid) anandamide is principally degraded by the integral membrane enzyme fatty acid amide hydrolase (FAAH). Pharmacological blockade of FAAH has emerged as a potentially attractive strategy for augmenting endocannabinoid signaling and retaining the beneficial effects of cannabinoid receptor activation, while avoiding the undesirable side effects, such as weight gain and impairments in cognition and motor control, observed with direct cannabinoid receptor 1 agonists. Here, we report the detailed mechanistic and pharmacological characterization of N-pyridazin-3-yl-4-(3-{[5-(trifluoromethyl)pyridin-2-yl]oxy}benzylidene)piperidine-1-carboxamide (PF-04457845), a highly efficacious and selective FAAH inhibitor. Mechanistic studies confirm that PF-04457845 is a time-dependent, covalent FAAH inhibitor that carbamylates FAAH's catalytic serine nucleophile. PF-04457845 inhibits human FAAH with high potency (k(inact)/K(i) = 40,300 M(-1)s(-1); IC(50) = 7.2 nM) and is exquisitely selective in vivo as determined by activity-based protein profiling. Oral administration of PF-04457845 produced potent antinociceptive effects in both inflammatory [complete Freund's adjuvant (CFA)] and noninflammatory (monosodium iodoacetate) pain models in rats, with a minimum effective dose of 0.1 mg/kg (CFA model). PF-04457845 displayed a long duration of action as a single oral administration at 1 mg/kg showed in vivo efficacy for 24 h with a concomitant near-complete inhibition of FAAH activity and maximal sustained elevation of anandamide in brain. Significantly, PF-04457845-treated mice at 10 mg/kg elicited no effect in motility, catalepsy, and body temperature. Based on its exceptional selectivity and in vivo efficacy, combined with long duration of action and optimal pharmacokinetic properties, PF-04457845 is a clinical candidate for the treatment of pain and other nervous system disorders.

Outcomes after carotid angioplasty and stenting in symptomatic octogenarians.

OBJECTIVE: Octogenarians were excluded from participation in many carotid endarterectomy trials due to the high complication rates observed in past studies. However, stroke resulting from carotid stenosis is expected to increase with the aging population. Moreover, advances in Carotid Angioplasty and Stenting (CAS) techniques have resulted in perceived improved safety of this procedure. We sought to review our experience with carotid stenting in symptomatic octogenarians with an emphasis on short-term outcomes and complications. METHODS: This is a retrospective longitudinal cohort study of all symptomatic patients who underwent CAS in our center between 1997 and 2007. Thirty-day stroke and death rates, and length of hospitalization were compared between the symptomatic octogenarians and non-octogenarians. RESULTS: A total of 214 procedures were performed on 211 symptomatic patients (56 females). Fifty-nine patients (14 females) were octogenarians. The median (interquartile range) age on procedure date for the octogenarian cohort was 83 (4) years. Periprocedural death occurred in two (3.4%) octogenarians and five (3.3%) non-octogenarians (p = 0.97). At 30 days from the procedure, stroke occurred in four (6.8%) octogenarians and seven (4.6%) non-octogenarians (p= 0.52). The mean hospital stay (4.8 days) was not different between the two cohorts. Age was not a predictor of the 30-day risk of composite stroke or death. CONCLUSION: The complications rate observed in octogenarians was not significantly higher than non-octogenarians. Our findings suggest that octogenarians should be included in randomized trials examining CAS to better define the risk-benefit profile of this procedure in the elderly.

Modulating target engagement of small molecules via drug delivery: approaches and applications in drug discovery and development.

Drug-delivery technologies for modified drug release have been in existence for decades, but their utilization has been largely limited to post-launch efforts improving therapeutic outcomes. Recently, they have gained renewed importance because the pharmaceutical industry is steadily shifting to a more integrated discovery-development approach. In discovery, modulating target engagement via drug-delivery technologies can enable crucial pharmacological studies for building well-defined criteria for molecular design. In development, earlier implementation of delivery technologies can enhance the value of drug products through reduced dosing frequency and improved tolerability and/or safety profile, thereby leading to better adherence and therapeutic effectiveness.

Carotid stenting in asymptomatic carotid stenosis: the Calgary experience.

BACKGROUND: Although carotid endarterectomy is considered the 'gold standard' for standard risk symptomatic patients, the treatment of choice for asymptomatic patients remains controversial. Carotid stenting has demonstrated real-world outcomes consistent with established guidelines for carotid endarterectomy in asymptomatic high-surgical risk patients in recent prospective multicenter trials. We describe our experience with asymptomatic patients who underwent carotid stenting at our center in a routine clinical setting. METHODS: This is a retrospective, longitudinal cohort study of patients who underwent carotid angioplasty and stenting at the Foothills Medical Center, Calgary, Canada between 1997 and 2007. The qualifying events were categorized as symptomatic and asymptomatic. The procedures were performed by four experienced neurointerventionists. The primary outcome was stroke or death at 30-day follow- up. RESULTS: 243 patients underwent 255 carotid stenting procedures. Their ages ranged from 50 to 83 years; the mean age was 72.0 ± 9.3 years; 67(26.3%) were women. Forty one patients (16.1%) were asymptomatic; 214 patients (83.9%) were symptomatic. The patients in the asymptomatic group were significantly younger - 66.0 ± 8.8 years compared to patients in the symptomatic group 73.2 ± 8.9 years (p < 0.0001). Intraprocedurally one minor stroke (2.4%) occurred in the asymptomatic group. At 30-day follow-up, no deaths or further strokes were noted in the asymptomatic group; while eight deaths, six major and seven minor strokes occurred in the symptomatic group (p = 0.22). CONCLUSION: Carotid stenting appears to be a safe procedure in asymptomatic patients with severe carotid stenosis in routine clinical settings as witnessed in this single center study.

Synthesis and characterization of Zinc oxide nanoparticles utilizing seed source of Ricinus communis and study of its antioxidant, antifungal and anticancer activity.

ZnO nanoparticles have been synthesized using solution combustion technique and its antioxidant, antifungal, anticancer activity was studied. Ricinus communis plant seed extract used as fuel in synthesis by the solution combustion technique. Powder X-ray diffraction (PXRD) demonstrates the arrangement of a crystalline hexagonal stage (ICDD card number 89-1397) with space aggregate P63mc (186) and cell parameters a = b = 3.253, c = 5.213 Å. The normal crystallite measure is 20 nm which is ascertained by Debye - Scherer's formula. The Purity of the sample and metal to oxygen bond development was affirmed by utilizing Fourier transformation infrared (FTIR) spectroscopy and the particle size and shape was confirmed by HRTEM. Antifungal action of ZnO NPs was studied against Aspergillus and Penicillium by well dispersion strategy. The antifungal activity shows that ZnO NPs constitute as an effective fungicidal agent against both Aspergillus (4 ±â€¯0.5 mm) and Penicillium (3 mm ±â€¯0.4 mm) at 30 µg/mL fixation. ZnO nanoparticles were subjected to antioxidant activity. The objective of the study was to analyze the anticancer property of ZnO NPs on MDA-MB 231 cancer cells. To check the efficacy of the synthesized drug ZnO NPs MTT assay was performed, that determines % viability and/or cytotoxicity. IC50 of ZnO NPs in case of MDA-MB-231 breast cancer was 7.103 µg/mL. Anticancer outcome demonstrates that ZnO NPs is active against in MDA-MB-231 cells.

Ruthenium-decorated vanadium pentoxide for room temperature ammonia sensing.

Layer structured vanadium pentoxide (V2O5) microparticles were synthesized hydrothermally and successfully decorated by a facile wet chemical route, with ∼10-20 nm sized ruthenium nanoparticles. Both V2O5 and ruthenium nanoparticle decorated V2O5 (1%Ru@V2O5) were investigated for their suitability as resistive gas sensors. It was found that the 1%Ru@V2O5 sample showed very high selectivity and sensitivity towards ammonia vapors. The sensitivity measurements were carried out at 30 °C (room temperature), 50 °C and 100 °C. The best results were obtained at room temperature for 1%Ru@V2O5. Remarkably as short a response time as 0.52 s @ 130 ppm and as low as 9.39 s @ 10 ppm recovery time at room temperature along with high selectivity towards many gases and vapors have been noted in the 10 to 130 ppm ammonia concentration range. Short response and recovery time, high reproducibility, selectivity and room temperature operation are the main attributes of the 1%Ru@V2O5 sensor. Higher sensitivity of 1%Ru@V2O5 compared to V2O5 has been explained and is due to dissociation of atmospheric water molecules on 1%Ru@V2O5 as compared to bare V2O5 which makes hydrogen atoms available on Brønsted sites for ammonia adsorption and sensing. The presence of ruthenium with a thin layer of oxide is clear from X-ray photoelectron spectroscopy and that of water molecules from Fourier transform infrared spectroscopy.

Hypertrophic cranial pachymeningitis in countries endemic for tuberculosis: diagnostic and therapeutic dilemmas.

Hypertrophic cranial pachymeningitis (HCP) is an uncommon disorder with few studies correlating clinical, imaging and histopathological features. The aim of this study was to describe clinical and laboratory observations and therapeutic options of patients with HCP. Eleven patients with HCP (M:F 6:5; age range, 23-52 years) were evaluated over 10 years. Etiology was ascertained by MRI and laboratory tests and confirmed by biopsy of meninges and/or brain (7), nasal mucosa (1), mediastinal lymph node (1), muscle (2) or conjunctiva (2). Salient clinical features were headache (7), multiple cranial neuropathies (8), visual disturbances (6), seizures (2) and hemiparesis (2). Abnormal tests included: rapid erythrocyte sedimentation rate (3), positive serum venereal disease testing (1), chest CT abnormalities (4/6) and positive Mantoux test (2/5). Cerebrospinal fluid changes (10/11) revealed the following: cell count 0-47/mm(3); protein 14-95 mg/mL; and glucose of 44-79 mg/mL. Contrast MRI revealed a variable extent of thickened dura mater in all patients. Histopathology (n=11) confirmed chronic inflammation (100%) and provided specific etiology in six (vasculitis [2], sarcoidosis [2], tuberculosis [1], Wegener's granulomatosis [1]). Treatment included steroids only (4), anti-tubercular therapy with steroids (5), penicillin (1) and cyclophosphamide and plasmapheresis (1). During follow-up (27.0+/-26.3 months) there was significant recovery (9/9). On serial imaging (4), the lesion remained the same in three and resolved partially in one patient. HCP, despite frequently posing diagnostic and therapeutic challenges, has favorable outcome when treated appropriately.

Solubility measurement of polymorphic compounds via the pH-metric titration technique.

In drug development, the thermodynamically most stable form of a compound is preferred because metastable forms are prone to transform to the stable form during processing, formulation, or storage [Guillory, J.K., 1999. Generation of polymorphs, hydrates, solvates, and amorphous solids. In: Brittain, H.G. (Ed.), Polymorphism in Pharmaceutical Solids. Marcel Dekker, New York, pp. 183-226]. It is therefore important to discover and characterize the stable form as early as possible. One of the most important properties to determine is thermodynamic solubility. However, due to compound and time constraints this solubility value is usually not determined until late in discovery. This report explores the ability of the pH-metric titration method to measure intrinsic solubility of the stable form of compounds that exist in one or more polymorphic forms. One metastable form and the stable form of eight compounds were examined. Intrinsic solubility was measured via pH-metric titration. The technique was performed on a larger scale in order to monitor polymorphic form changes by powder X-ray diffraction. Shake-flask solubility and corresponding X-ray diffraction data of each form was also determined. The results of this study indicate that, in general, when starting with a metastable polymorph, the pH-metric titration method is able to achieve the solubility of the stable form by the third titration, while the traditional shake-flask solubility method is unable to consistently determine the stable form solubility.

Automated approach to couple solubility with final pH and crystallinity for pharmaceutical discovery compounds.

The design and validation of a novel high-throughput system for thermodynamic solubility determination requiring only 5 mg of sample is described. The system uses a sintered nickel filter assembly to recover excess solids from saturated solutions for rapid crystallinity assessment via powder X-ray diffraction (PXRD). Moreover, the system measures the pH of filtrates to provide a final pH value with the solubility measurement. The limit of detection for the UV-vis plate reader used on this system is approximately 0.001 mg/ml, while the practical upper limit is approximately 3 mg/mL. The solubility measurements of 60 proprietary Pfizer compounds were used to validate the nickel filter assembly against a more conventional polyvinylidenedifluoride (PVDF) filter. Additionally, a comparison was made between a subset of 10 compounds run on the automated system and a more traditional shake-flask method employing HPLC analysis. In both cases, a favorable comparison was obtained.

Solubility: it's not just for physical chemists.

Solubility data are used to make crucial decisions from the earliest stages of drug discovery throughout the development process, but often the decision-maker is far removed, in terms of both organization and scientific background, from the scientist who generates the data. Here we provide a reference point for consumers of solubility who are presented with increasingly sophisticated strategies to measure sooner, faster or more accurately. We discuss the fundamental forces that govern solubility, the role of physical-chemical parameters such as pH and pK(a), and the principles involved in different solubility measurements. Our ultimate goal is to enable a decision-maker, when presented with solubility data, to have in hand the tools to evaluate not just the magnitude but also the context and appropriateness of those measurements to the drug in question.

Evaluation of the chemiluminescent nitrogen detector for solubility determinations to support drug discovery.

Solubility measurements using chemiluminescent nitrogen detection (CLND) has advantages of reduced compound requirement and increased throughput compared to UV-spectrophotometric and HPLC-based measurements. CLND with direct flow injection was evaluated for the measurement of thermodynamic solubility to support drug discovery. The limit of quantitation (LOQ), accuracy, and day-to-day reproducibility of the detector were measured. Measurements made on CLND were compared to those obtained from UV spectrophotometry and HPLC. Based on the results obtained, it was concluded that the CLND performs satisfactorily for discovery stage thermodynamic solubility measurements.

Diabetic nonketotic hyperosmolar state: interesting imaging observations in 2 patients with involuntary movements and seizures.

We report two patients of diabetic nonketotic hyperosmolar state presenting acutely with "self-limiting hemichorea - hemiballismus" and "generalized convulsive status epilepticus". CT scan in both the patients revealed a hyperdense nonenhancing basal ganglia. Magnetic resonance imaging brain of patient 1 showed it to be hyperintense on T1W image and iso-hyper intense on T2W image, minimally enhancing with contrast injection.

Discovery and Characterization of 2-Acylaminoimidazole Microsomal Prostaglandin E Synthase-1 Inhibitors.

As part of a program aimed at the discovery of antinociceptive therapy for inflammatory conditions, a screening hit was found to inhibit microsomal prostaglandin E synthase-1 (mPGES-1) with an IC50 of 17.4 µM. Structural information was used to improve enzyme potency by over 1000-fold. Addition of an appropriate substituent alleviated time-dependent cytochrome P450 3A4 (CYP3A4) inhibition. Further structure-activity relationship (SAR) studies led to 8, which had desirable potency (IC50 = 12 nM in an ex vivo human whole blood (HWB) assay) and absorption, distribution, metabolism, and excretion (ADME) properties. Studies on the formulation of 8 identified 8·H3PO4 as suitable for clinical development. Omission of a lipophilic portion of the compound led to 26, a readily orally bioavailable inhibitor with potency in HWB comparable to celecoxib. Furthermore, 26 was selective for mPGES-1 inhibition versus other mechanisms in the prostanoid pathway. These factors led to the selection of 26 as a second clinical candidate.

HIV with non-communicable diseases in primary care in Kibera, Nairobi, Kenya: characteristics and outcomes 2010-2013.

BACKGROUND: Antiretroviral therapy (ART) has increased the life expectancy of people living with HIV (PLHIV); HIV is now considered a chronic disease. Non-communicable diseases (NCDs) and HIV care were integrated into primary care clinics operated within the informal settlement of Kibera, Nairobi, Kenya. We describe early cohort outcomes among PLHIV and HIV-negative patients, both of whom had NCDs. METHODS: A retrospective analysis was performed of routinely collected clinic data from January 2010 to June 2013. All patients >14 years with hypertension and/or diabetes were included. RESULTS: Of 2206 patients included in the analysis, 210 (9.5%) were PLHIV. Median age at enrollment in the NCD program was 43 years for PLHIV and 49 years for HIV-negative patients (p<0.0001). The median duration of follow up was 1.4 (IQR 0.7-2.1) and 1.0 (IQR 0.4-1.8) years for PLHIV and HIV-negative patients, respectively (p=0.003). Among patients with hypertension, blood pressure outcomes were similar, and for those with diabetes, outcomes for HbA1c, fasting glucose and cholesterol were not significantly different between the two groups. The frequency of chronic kidney disease (CKD) was 12% overall. Median age for PLHIV and CKD was 50 vs 55 years for those without HIV (p=0.005). CONCLUSIONS: In this early comparison of PLHIV and HIV-negative patients with NCDs, there were significant differences in age at diagnosis but both groups responded similarly to treatment. This study suggests that integrating NCD care for PLHIV along with HIV-negative patients is feasible and achieves similar results.

Dissolution testing of a poorly soluble compound using the flow-through cell dissolution apparatus.

Dissolution of Pfizer Compound PD198306, a poorly soluble compound, was studied in 25 mM pH 9 sodium phosphate solution with 0.5% SLS using the flow-through cell dissolution apparatus. Unmicronized and micronized drug powders were tested. Several methods of loading the drug powder into the flow-through dissolution cells and their impact on dissolution were investigated. The influence of flow rate of the dissolution medium on the rate and extent of dissolution were studied. PD198306 has poor wettability even in the presence of 0.5% SLS. It was found that loading the drug powder into the dissolution cell in the form of a suspension provided the best dissolution profile in terms of the rate and extent of dissolution. The flow rate of 4 ml/min resulted in good particle size discrimination.