Losartan: Benefits Beyond Hypertension

Losartan was the first angiotensin AT1 receptor blocker (ARB) approved by US Food and Drug Administration (FDA) for the treatment of hypertension. In addition to its established antihypertensive and end organ effects, several benefits of losartan beyond its antihypertensive effect have been demonstrated in clinical trials. Apart from its class effects of ARBs, losartan has pharmacokinetic and pharmacodynamic properties that are unique to it. It has shown considerable benefits as uricosuric agent, in erectile dysfunction and in prevention of stroke in hypertension patients with left ventricular hypertrophy. This review presents the benefits of losartan beyond being a hypertensive agent and associated clinical outcomes

ISOLATION AND IDENTIFICATION OF A NEW PHYTOSTEROL FROM HOLOPTELEA INTEGRIFOLIA (ROXB) PLANCH LEAVES

Objective: The Plant Holoptelea Intergrifolia (Roxb.) Planch is being used for the treatment of various disorders since time immemorial in the indigenous system of medicine in India. The main objective of the work was to isolate a new phytosterol from petroleum ether extract of Holoptelea  Integrifolia leaves using Preparative Thin Layer Chromatography (TLC). Methods: As per ICH guidelines we have Prepared Thin Layer Chromatographic plates for separation of a new phytosterol from Petroleum  ether extract of leaves of Holoptelea  Intergrifolia (Roxb.) Planch. The mobile phase used for separation of phytosterol consisted ofChloroform: Ethyl acetate, in the volume ratio of 4:6 (v/v), UV, LC/MS, IR and NMR spectral analytical techniques were used for identification and confirmation of structure of a new Phytosterol by Preparative TLC. Results: Preliminary phytochemical analysis of petroleum ether extract of Holoptelea integrifolia leaves showed the presence of steroids, terpenoids, alkaloids, glycosides, flavonoids, proteins, tannins and carbohydrates. The isolated phytosterol designated as17-(6-(diethylamino) decan-3-yl)-10,13-dimethyl-12,13-dihydro-10H-cyclopenta[a] phenanthren-3-ol. It responded positively to Liebermann Burchard test indicating steroidal nature of the molecule. Conclusion: On the basis of spectral data analysis and chemical reactions, the structure of a new phytosterol isolated by preparative TLC from petroleum ether extract of leaves of Holoptelea Integrifolia (Roxb.) Planch has been formulated by UV, LC/MS, IR and NMR spectral analysis as 17-(6-(diethylamino) decan-3-yl)-10,13-dimethyl-12,13-dihydro-10H-cyclopenta[a] phenanthren-3-ol This is a new phytosterol isolated from plant source and being reported for the first time

REVERSAL OF CLONIDINE-INDUCED HYPOTHERMIA BY DECAFFEINATED TEA/COFFEE EXTRACT, AND THEIR FRACTIONS IN MICE

Objective: To study the effect of decaffeinated tea extract (DTE) and decaffeinated coffee extract (DCE) and their respective fractions viz: chloroform fractions (DTCf and DCCf), ethyl acetate fractions (DTEa and DCEa), diethyl ether fractions (DTDe and DCDe) and acetone-water fractions (DTAw and DCAw) against clonidine-induced hypothermia in mice. Methods: Clonidine (0.1 mg/kg, i. p.) administered to a group of mice pretreated 30 min before with the dose of DTE or DCE or their respective fractions. Rectal temperature was measured at the time of clonidine administration and thereafter at every 30 min up to 2 h test period. Results: DTE 200 DTE 300 has significantly inhibited clonidine-induced hypothermia. Among the fractions tested, DTE fraction-DTEa 100 and 200 and DCE fractions DCDe 200 and DCAw 200 significantly (p<0.0001) reversed clonidine-induced hypothermia; the effect of DTEa was found to be more sustained. Conclusion: Both, the decaffeinated tea and coffee contain ingredients that reverse clonidine-induced hypothermia, but they are required to do so in very large doses which are not achievable with normally administered doses of decaffeinated tea or coffee

Bioanalytical Method Development and Validation: A Review

For various types of drug approval processes like INDs, NDAs, ANDAs, veterinary drug approval, the data related to bioanalytical method development and validation is needed to sponsors. Various agencies namely US FDA, American association of pharmaceutical scientists (AAPS), Health protection Branch (HPB), Association of analytical chemists (AOAC), Center for Veterinary Medicine (CVM), U.S. Department of Health and Human Services Food and drug Administration, Center for Drug Evaluation and Research (CDER), European Medicine Agency (EMA), China Food and Drug administration(CFDA), European Bioanalytical Forum (EBF), Global CRO council (GCC), ANVISA (Brazil), Japan Bioanalytical Forum (JBF) had done collective efforts at different timings to regulate and harmonize bioanalytical method development and validation

Dapagliflozin in the Landscape of Type 2 Diabetes Management

As per current statistics, India accounts for more than 74 million individuals living with diabetes. Many of these individualshave associated cardiovascular disease (CVD) and chronic kidney disease (CKD) comorbidities. Optimal glycemic managementis important because uncontrolled glycemia may accelerate the macrovascular and microvascular complications, furtheraggravating the comorbid conditions. Metformin is used as the first-line therapy in most persons. However, there are somewho do not tolerate metformin, are unable to achieve required glycemic targets or require greater efforts for cardiovascular(CV) risk reduction. These patients require an alternative hypoglycemic agent to be used as either monotherapy or ascombination treatment with metformin, respectively. Sodium-glucose cotransporter-2 (SGLT2) inhibitors are one such novelclass of drugs that can be used as either monotherapy or as part of two drug (dual) or three drug (triple) combinations withother oral hypoglycemic agents or insulin. Dapagliflozin is a promising option for managing type 2 diabetes with CV andrenal benefits, weight and blood pressure reducing properties. A low risk of hypoglycemia and drug-drug interactions are theadded advantages. In this article, the authors have reviewed the existing clinical evidences on dapagliflozin and highlightedits place in the diabetes management landscape

Essential Oil of Myrtus communis Inhibits Inflammation in Rats by Reducing Serum IL-6 and TNF-α:

The topical anti-inflammatory activity of the essential oil of Myrtus communis L. was studied using croton oil induced ear edema and myeloperoxidase (MPO) activity in mice, and cotton pellet induced granuloma, and serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in rats. On topical application, the oil exhibited a significant decrease in the ear edema as well as MPO activity. The oil also inhibited cotton pellet-induced granuloma and serum TNF-α and IL-6. It can be concluded that the essential oil of Myrtus communis reduces leukocyte migration to the damaged tissue and exhibits anti-inflammatory activity

Advances in Sardinian Withania somnifera (L.) Dunal crops through phytochemical and biological approaches

Withania somnifera (L.) Dunal is widely used in the Indian traditional system of medicine to promote general health, wellness, and longevity. Its pharmacological properties are attributed to a group of molecules called withanolides, among which Withaferin A holds great interest for its anti-carcinogenic action. For this reason, numerous studies in recent years have focused on different metabolic or genetic engineering solutions to increase its yield. Here, we present the Sardinian chemotype of Withania somnifera as a potential crop for the extraction of Withaferin A. W. somnifera was cultivated from Sardinian wild germplasm collected in the northeast of the island. After 18 months, the leaves and the roots were collected and their methanolic extract was analyzed by HPLC. 0.3 mg/g DW of Withanolide A (WA), 1.0 mg/g DW of Withanolide B (WB) and 17.7 mg/g DW of Withaferin A (WF) were detected in the leaf sample, while lower values were detected in the roots (0.1 mg/g WF, 0.3 WA mg/g, 0.1 mg/g WB, 0.2 mg/g WO). This research not only confirms the high Withaferin A content found in the wild population leaves, but shows how they are reproducible in cultivated specimens, highlighting Sardinian W. somnifera leaves as a potential source of high-content Withaferin A products. Finally, we focused on the leaves extract by characterizing the phenolic and flavonoid content, as well as the in-vitro antioxidant capacity by DPPH and ABTS assays, revealing a significant amount of phenolic compounds and a related free radical scavenging activity. The leaves extract was further characterized for its anti-aging properties for potential cosmetic application, by the inhibition of tyrosinase, elastase, and collagenase enzymes

Inhibition of Morphine- and Ethanol-Mediated Stimulation of Mesolimbic Dopamine Neurons by Withania somnifera

Morphine- and ethanol-induced stimulation of neuronal firing of ventral tegmental area (VTA) dopaminergic neurons and of dopamine (DA) transmission in the shell of the nucleus accumbens (AcbSh) represents a crucial electrophysiological and neurochemical response underlying the ability of these compounds to elicit motivated behaviors and trigger a cascade of plasticity-related biochemical events. Previous studies indicate that the standardized methanolic extract of Withania somnifera roots (WSE) prevents morphine- and ethanol-elicited conditioned place preference and oral ethanol self-administration. Aim of the present research was to investigate whether WSE may also interfere with the ability of morphine and ethanol to stimulate VTA dopaminergic neurons and thus AcbSh DA transmission as assessed in male Sprague- Dawley rats by means of patch-clamp recordings in mesencephalic slices and in vivo brain microdialysis, respectively. Morphine and ethanol significantly stimulated spontaneous firing rate of VTA neurons and DA transmission in the AcbSh. WSE, at concentrations (200–400 mg/ml) that significantly reduce spontaneous neuronal firing of VTA DA neurons via a GABAA- but not GABAB-mediated mechanism, suppressed the stimulatory actions of both morphine and ethanol. Moreover, in vivo administration of WSE at a dose (75 mg/kg) that fails to affect basal DA transmission, significantly prevented both morphine- and ethanol-elicited increases of DA in the AcbSh. Overall, these results highlight the ability of WSE to interfere with morphine- and ethanolmediated central effects and suggest a mechanistic interpretation of the efficacy of this extract to prevent the motivational properties of these compounds

Protein folding and ER-export of neurotransmitter transporters in Drosophila melanogaster

Neurotransmittertransporter dienen dazu, freigesetzte Neurotransmitter wieder in die präsynaptische Nervenendigungen zurückzuholen. Gemeinsam mit den vesikulären Monoamintransporter, die die Neurotransmitter in den Vesikeln konzentrieren, erhalten sie die synaptische Homöostase. Wie alle Membranproteine, werden Neurotransmittertransporter der SLC6 Genfamilie werden im endoplasmatischen Retikulum (ER) synthetisiert. Nach abgeschlossener Faltung, rekrutieren die Transporteroligomere die Komponenten der COPII-Maschinerie für den ER-Export, sie werden in der Folge im ER-Golgi intermediären Kompartiment (ERGIC) and im Golgi sortiert und zur Plasmamembran dirigiert. Vor dem ER-Export unterliegen die Transporter einer stringenten Qualitätskontrolle im ER, sodass nur die nativ gefalteten Transporter das ER verlassen können. Wenn eine Punktmutation die Proteinfaltung beeinträchtigt, bleiben die Transport im ER stecken.^ ^Mehrere Erkrankungen lassen sich auch Punktmutationen von SLC6 Transporten zurückführen. Die Proteinfaltung kann durch chemische und pharmakologische Manipulationen unterstützt werden. In der vorliegenden Dissertation wurde eine Mutante des Drosophila melanogaster Dopamintransporters (dDAT-G108Q) als konzeptioneller Beweis für eine Pharmakochaperon-basierte Therapie untersucht. Fliegen, die diese Mutation tragen, haben ein stark reduzierte Schlafdauer; der Phänotyp entspricht dem DAT-defizienter fumin (=japanisch für schlaflos) Fliegen.^ Die Experiemnte zeigten, dass (i) dDAT-G108Q und sein humanes Äquivalent hDAT-G140Q im ER retiniert wurden, (ii) die heterelogous exprimierten Proteine die Zelloberfläche erreichten, wenn die Zellen mit Noribogain, einem DAT-Hemer, und mit Pifithrin-, einem Inhibitor des Hitzeschockproteins-70 (HSP70) inkubiert wurden, (iii) in Fliegen, die transgen für hDAT-G140Q waren, das Protein auf das neuronale Soma beschränkt war, aber die präsynaptischen Endigung erreichte, wenn den Fliegen über ihr Futter Noribogain verabreicht wurde; (iv) die Behandlung mit Noribogain, Pifithrin- oder deren Kombination normalisierte die Schlafdauer von Fliegen mit dDAT-G108Q aber nicht bei DAT-defizienten fumin Fliegen. Zusätzlich wurde die Leistungsfähigkeit der Drosophila-Genetik genutzt, um das Schicksal von Transportern zu untersuchen, die nach abgeschlossener Faltung den COPII-Mantel nicht rekrutieren können.^ SEC24 ist die COPII- Komponente, die als Cargo-Rezeptor für Proteinklienten dient. Bei Säugern liegen vier SEC24 Isoformen vor, bei Drosophila zwei. Serotonintransporter (SERT) von Säugern rekrutiert SEC24C für seinen ER-export über ein konserviertes, C-terminales R607I608-Motif. Hier wurde das Schicksal von hSERT-R607A untersucht: (i) diese Mutante verlässt das ER unabhängig von SEC24C; (ii) der SEC24C-unabhängige Export führt zur Anreicherung des Proteins im somatodendritischen Kompartiment von Neuronen, die aus den serotonergen Raphe-Kernen gewonnen. (iii) In Fliegen mit der äquivalenten Mutation im Drosophila SERT (dSERT-R509A) ereicht das Protein nicht das axonale Kompartiment. Diese Befunde bestätigen, dass der SEC24-abhängige Export für die korrekte Verteilung von SERT notwendig ist. Sie legen auch einen experimentellen Zugang nahe, mit dem die biologische Rolle von SERT im somatodendritischen Kompartiment geprüft werden kann.Neurotransmitter transporters retrieve the neurotransmitter from synaptic and extrasynaptic places and work in relay with vesicular monoamine transporters to maintain synaptic homeo-stasis. Neurotransmitter transporters belonging to SLC6 gene family begins their journey by folding in the endoplasmic reticulum (ER). Upon achieving the natively folded state the oligo-merized transporters recruit components of the COPII machinery for ER-export. The transporters are further sorted in the ER-Golgi intermediate compartment (ERGIC) and the Golgi, before reaching the plasma membrane. Before exiting the ER transporters undergo stringent ER quality control and only the natively folded protein exits the ER. In rare circum-stances where a point mutation affects the protein folding, transporters remain stalled in the ER. A number of diseases result from point mutations in the SLC6 family. Chemical and pharmacological means can be used can to assist protein folding.^ ^In the current thesis, a mutant of the Drosophila melanogaster dopamine transporter (dDAT-G108Q) was studied as a proof-of-concept. Flies carrying this mutation show a sleepless phenotype, similar to fumin (=DAT-deficient) flies. The present thesis showed that (i) - as a consequence of the mutation - dDAT-G108Q and its human counterpart hDAT-G140Q were retained in the ER; (ii) the heterelogously expressed proteins reached the cell surface, if the cells were incubated with noribogaine, an inhibitor of DAT, and pifithrin-, an inhibitor of heat shock protein-70 (HSP70); (iii) in flies harboring hDAT-G140Q, the protein was confined to the neuronal soma but was delivered to the presynaptic specializations, if the flies had been administered noribo-gaine via their food; (iv) similarly, treatment with noribigaine, pifithrin- or a combination of the two compounds restored sleep to flies harboring dDAT-G108Q but not to DAT-deficient fumin flies.^ In addition, I used the power of Drosophila genetics to study the fate of transpor-ters, which fail to recruit the COPII coat after they have reached the folded state. SEC24 is the COPII component, which acts as cargo receptor for client proteins. In mammals and Droso-phila, there are four and two SEC24 isoforms, respectively. Mammalian serotonin transpor-ters (SERT) specifically recruit SEC24C for ER-export via the conserved R607I608-motif in the C-terminus. In the current thesis, the fate of the SERT-R607A mutant was examined: (i) hSERT-R607A exits the ER in a manner independent of SEC24C; (ii) SEC24C-independent export results in enrichment of the protein in somatodendritic compartment of cultured raphe neurons. (iii) In flies, harboring the equivalent mutation in Drosophila SERT (dSERT-R509A), the protein does not enter the axonal compartment. These observations confirm that SEC24-dependent export is required for correct targeting of SERT.^ They also point to an approach to explore the biological role of SERT in the somatodendritic compartment.submitted by Ameya Sanya KastureMedizinische Universität Wien, Diss., 201