Validated LC-MS/MS Method for Simultaneous Quantitation of SAFit-1 and SAFit-2 in Mice Plasma: Application to a Pharmacokinetic Study.

SAFit-1 and SAFit-2 are selective FKBP51 (FK506-binding protein 51) ligands. In this paper, we present the development and validation data of an LC-MS/MS method for the simultaneous quantitation of SAFit-1 and SAFit-2 in mice plasma as per FDA regulatory guideline. SAFit-1 and SAFit-2 along with internal standard were extracted from mice plasma using liquid-liquid extraction method. Chromatographic resolution of SAFit-1, SAFit-2 and the internal standard (warfarin) was achieved on an X-Terra phenyl column using 0.2% formic acid:acetonitrile (20:80, v/v) as an eluent, which was delivered at a flow-rate of 0.9 mL/min. The MS/MS ion transitions monitored were m/z 748.4→420.4, 803.7→384.3 and 309.2 →163.2 for SAFit-1, SAFit-2 and the internal standard, respectively. The linearity range was 2.45-2446 ng/mL for both SAFit-1 and SAFit-2. The intra- and inter-day accuracy and intra- and inter-day precision were in the range of 0.90-1.07 and 2.38-10.8%, respectively for SAFit-1; 0.97-1.15 and 0.23-12.5%, respectively for SAFit-2. Both SAFit-1 and SAFit-2 were found to be stable in stability studies (up to three freeze-thaw cycles and for long-term at -80°C for 30 days) and processed (bench-top for 3 h and in in-injector for 16 h) samples. The application of the validated method was shown in a pharmacokinetic study in mice.

Design and synthesis of a novel series of cyanoindole derivatives as potent γ-secretase modulators.

The discovery, design and synthesis of a new series of GSMs is described. The classical imidazole heterocycle has been replaced by a cyano group attached to an indole nucleus. The exploration of this series has led to compound 26-S which combined high in vitro and in vivo potency with an acceptable drug-like profile.

Novel KV7 ion channel openers for the treatment of epilepsy and implications for detrusor tissue contraction.

Neuronal voltage-gated potassium channels, KV7s, are the molecular mediators of the M current and regulate membrane excitability in the central and peripheral neuronal systems. Herein, we report novel small molecule KV7 openers that demonstrate anti-seizure activities in electroshock and pentylenetetrazol-induced seizure models without influencing Rotarod readouts in mice. The anti-seizure activity was determined to be proportional to the unbound concentration in the brain. KV7 channels are also expressed in the bladder smooth muscle (detrusor) and activation of these channels may cause localized undesired effects. Therefore, the impact of individual KV7 isoforms was investigated in human detrusor tissue using a panel of KV7 openers with distinct activity profiles among KV7 isoforms. KCNQ4 and KCNQ5 mRNA were highly expressed in detrusor tissue, yet a compound that has significantly reduced activity on homomeric KV7.4 did not reduce detrusor contraction. This may suggest that the homomeric KV7.4 channel plays a less significant role in bladder contraction and further investigation is needed.

Assessment of Drug Usage Pattern in Patients Treated for Vernal Keratoconjunctivitis Attending a Tertiary Eye Care Centre in Eastern India: A Cross-Sectional Study.

INTRODUCTION: Vernal keratoconunctivitis is a recurrent, seasonal, allergic condition affecting children and adolescents in warmer regions worldwide. OBJECTIVE: To assess the drug usage pattern for the management of vernal keratoconjunctivitis (VKC). MATERIALS & METHODS: This was a 12 weeks long hospital based cross sectional study conducted in the outpatient unit of the department of ophthalmology of a teaching hospital. It included all consecutive patients diagnosed with VKC who satisfied the inclusion criteria. Patients underwent clinical examination and the severity of their disease was graded. Their prescriptions were scanned for duration of therapy, total number of medications, route of administration, the frequency of dosage and change in medications if any, generic names of drugs being prescribed or not and all demographic parameters were recorded in a suitable record form. A p value of <0.05 was considered to be statistically significant. RESULTS: 248 patients were enrolled in this study of whom the majority were male (172). The average age of the patients was 8.27 years (SD ± 3.02 years). The mean duration of disease was 15 months (SD ± 9.13 years). The greatest number of children belonged to severity grade 1 (27.82%). The total number of drugs prescribed in this study was 583 with an average of 2.26 drugs per encounter. The commonest prescribed drugs were topical anti-allergics (26.75%) followed closely by lubricants (25.73%) and topical steroids (21.42%). CONCLUSION: Anti allergics and lubricants are the mainstays in the management of VKC. The current study is reflective of this. Clearer guidelines need to be formulated for the better and rational management of VKC.

Validated LC-MS-MS Method for Determination of SF0034 in Mice Plasma: Application to a Pharmacokinetic Study in Mice.

A rapid and sensitive assay method has been developed and validated using liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the positive-ion mode for the estimation of SF0034 in mice plasma. The assay procedure involves a simple protein precipitation of SF0034 and tolbutamide (internal standard, IS) from mice plasma. Chromatographic separation was performed on an Atlantis dC18 column using an isocratic mobile phase comprising 0.2% formic acid:acetonitrile (10:90, v/v) at a flow rate of 0.60 mL/min. The total run time was 2.5 min. For mass spectrometric detection, the multiple reaction monitoring was used and ion transitions monitored were m/z 322 → 248 for SF0034 and 271 → 155 for IS. Method validation was performed as per regulatory guidelines and the results met the acceptance criteria. A calibration curve was constructed in the range of 2.08-2,078 ng/mL. The intra- and inter-day precision was in the range of 1.06-14.4% and 7.16-11.7%, respectively. This novel method has been applied to a pharmacokinetic study in mice.

Penetrating ocular fish hook injury: a case report.

BACKGROUND: Fishing is a popular rural recreational activity. Fortunately, penetrating ocular injuries with fish hooks are rare. These injuries are usually caused by fish hook prongs penetrating the ocular tissues. We report a rare case of penetrating fish hook injury to the globe and its successful surgical management. CASE: A 12- year-old female child was referred to the causality with a fish hook embedded in her right eye. She sustained the injury while fishing with her father. The fish hook had penetrated the globe obliquely. The hook was removed via its entrance wound under general anesthesia. Subsequently the child developed traumatic cataract which was operated with a final visual outcome of 6/12. CONCLUSION: Visual prognosis can be profoundly affected by the initial management. Prompt surgical intervention as done in our case is recommended to prevent significant visual loss. Keywords Fishhook; penetrating ocular injury; traumatic cataract.

Bilateral central serous chorio-retinopathy in pregnancy presenting with severe visual loss.

INTRODUCTION: Bilateral central serous chorio-retinopathy is an unusual presentation. OBJECTIVE: To report a case of pregnancy-induced bilateral central serous chorio-retinopathy. CASE: A thirty-year-old woman complaining of reduced vision in both the eyes was referred for ophthalmoscopic evaluation in her third trimester of pregnancy. The best-corrected visual acuity was 6/60 in the right eye and finger counting at four meters in the left. The funduscopic examination revealed serous macular detachment with white sub-retinal exudates in both the eyes. Optical coherence tomography sections through the macula demonstrated serous elevation of the retina and retinal pigment epithelial detachment in both the eyes. The serous macular detachment resolved and the patient recovered to the visual acuity of 6/6 in both the eyes after twelve weeks of delivery. The OCT showed resolution of the sub-retinal fluid and disappearance of the sub-macular exudate in the right eye, but the resolution was partial in the left eye. CONCLUSION: Pregnancy is a risk factor for CSCR and it can present with severe bilateral visual diminution.