Application of Elements of Quality by Design to Development and Optimization of HPLC Method for Fingolimod

Purpose: A HPLC method for Fingolimod was developed using a Quality by Design concept. QbD has gained importance in recent times due to regulatory requirements. Actual study was started after determination of target profile and qualification of instrument. Methods: Separation was carried on a Grace C-8 column (4.6 x 250 mm, 5-μm particle size).The composition of mobile phase was methanol and 20 mM ammonium formate buffer of pH5.8 in gradient mode HPLC method development is affected by critical factors like pH, flow rate and mobile phase composition. Results: To study the effects of these three factors on USP tailing, Box Behnken optimization model was applied. Desirability of the model was set at Tailing less than 1.2.Analysis of results was done using surface diagrams. Verification of Software generated results was done by taking six replicates of the run. Thus developed and optimised method was Finally validated as per ICH guideline. Conclusion: A Quality by Design approach has been successfully utilised in method development of the Fingolimod in bulk. All key aspect of QbD were tried to be implemented in said study. Systematic approach was utilized for method development which includes beginning with determination of target profile characteristics, instrument qualification, risk assessment, design of experiment and validation. Three factors i.e. Ph, flow rate and methanol concentration were analysed for their effect on USP tailing as a responce. Interaction and quadratic effect of the factors were studied with least possible runs by using Box Behnken model. Response surface diagrams and contour plots were studied for coming to conclusion which factors are affecting response and their limits were recorded. Optimum run condition was obtained; Replicates of run having optimized condition were taken to confirm the predicted response with actual response.

Abstract P473: Perfusion Imaging Identifies Patients With Mild Deficits Due to Large Vessel Occlusion Who May Benefit From Endovascular Thrombectomy: A Pooled International Cohort Study

Background: Efficacy and safety of endovascular thrombectomy (EVT) in large vessel occlusion (LVO) patients with mild deficits is unclear. Methods: Pooled cohort of pts with mild deficits (NIHSS<6) due to (ICA, M1, M2) LVO from EXTEND IA TNK I & II RCTs and prospective data from 12 centers (US, AUS, NZ, Canada, Spain) from 1/2013 to 2/2020 was divided into medical management (MM) vs EVT. All pts had baseline CT, CTA, CTPRAPID software estimated ischemic core and mismatch. Pts stratified into with or without target profile (≥1cc core / mismatch ratio ≥ 1.8 / mismatch volume ≥ 15cc). Primary outcome- excellent (90 day mRS 0-1); Secondary- mRS shift, safety (sICH, neuro-worsening, mortality). Results: Of 371 pts, 189 (51%) had EVT. Time LKW to EVT center: EVT 165 (70- 416) vs MM 200 (72-564) min, p=0.35 were similar. EVT pts had larger perfusion lesions (51 cc (23-86) vs 30.1 (5, 65), p<0.001), higher NIHSS 4 (2-5) vs 3 (2-4), p=0.009), less IV tPA (30% vs 41%, p=0.044), more M1s (44% vs 29%, p<0.001). 93 pts (25%) had target profile, of whom 60% had EVT. Of 278 without target profile, 48% had EVT. Among all pts, excellent outcomes and mRS distribution were similar (EVT 63.5% vs MM 59.1%, aOR 1.55, 95%, p=0.16) and (adj cOR 1.44, p=0.16) Fig 1A. EVT had worse safety; sICH (6% vs 0%, p=0.002); neuro-worsening (19% vs 3%, p<0.001) and mortality (5% vs 1%, p=0.06). With target profile, EVT associated with more excellent outcomes (66% vs 49%, aOR 4.44, 95% CI 1.04-18.95, p=0.04), shift to better outcomes (adj cOR 2.9, 95% CI 1.03-7.91, p=0.04) Fig 1B. Safety was similar; sICH 2% vs 0%, p>0.99, neuro-worsening 17% vs 6%, p=0.30) and mortality 5% vs 3%, p>0.99). Without target profile, excellent outcomes were similar without a shift, Fig 1C. Safety was worse with EVT: sICH 8% vs 0%, p=0.001; neuro-worsening 20% vs 3%, p<0.001). Conclusion: EVT was not associated with improved outcomes in patients with mild deficits; safety was worse. However, EVT was safe and associated with improved outcomes in target profile patients.

Classifying Car Owners in Latent Psychographic Profiles

Policy makers in urban areas are subjected to increasing pressure to find sustainable solutions to congestion and transportation. A detailed understanding of the motivations of car owners is required to enable the development of policies that are both socially fair and take effective measures. The objective of this study is to provide a more granular differentiation of car owners using psychographic profiles in three basic dimensions (privacy, autonomy, and car excitement). These profiles are also examined in relation to general travel behavior in everyday and long-distance travel. Data was collected in Munich and Berlin (Germany) and a latent class analysis was applied to segment respondents into latent profile classes. On this basis, six different profile classes were identified. In addition to the Car Independents profile class which does not have strong orientations toward the car, several profile classes were also identified with high concerns about “privacy” in relation to social distances in public transit. The information and analysis presented enables a deeper understanding of the motivations of the different target profile classes and discusses the need for tailored, socially fair measures to reduce car ownership and use within these groups.

The Relationship Between Penumbral Tissue and Blood-Brain Barrier Disruption in Acute Stroke Patients Presenting in an Extended Time Window

Background: Penumbral brain tissue identified with multimodal imaging can be salvaged with reperfusion in an extended time window. The risk of severe hemorrhagic complications after reperfusion therapy increases with worsening disruption of the blood-brain barrier (BBB). The relationship between penumbral tissue and BBB disruption has not been previously studied.Methods: Stroke patients presenting in an extended time window without a large vessel occlusion who underwent diffusion-perfusion MRI within 24 h of last-seen-normal were included. The volume of penumbral tissue was calculated using mismatch on MRI. Mean permeability derangement (MPD) of the BBB was measured within the ischemic lesion. A target profile (TP) for treatment was defined based on the EXTEND trial.Results: 222 patients were included with a median age of 73 and 55% women. The median NIHSS was 6, the mean core volume was 14 ml, the mean ischemic volume was 47 mL and the mean mismatch volume was 33 mL. Higher MPD was significantly associated with less mismatch volume (p = 0.001). A target profile was associated with lower MPD (OR 0.97; CI 0.96:0.99; p &lt; 0.001). Of the 105 patients who had a TP, 31 (30%) had a MPD &gt; 20% suggesting an increased risk of hemorrhage. Thus, 33% (74/222) of patients had a favorable profile for benefit and safety.Conclusions: Patients presenting in an extended time window with a favorable penumbral profile for treatment have less severe BBB disruption. Up to a third of patients who currently go untreated could be considered for enrollment in a clinical trial of thrombolysis in an extended time window.

The Persnickety Pervasiveness of Rating Enhancement in Personality Assessment

The current paper explores whether self and observer reports of personality are properly viewed through a contrasting lens (as opposed to a more consonant framework). Specifically, we challenge the assumption that self-reports are more susceptible to certain forms of response bias than are informant reports. We do so by examining whether selves and observers are similarly or differently drawn to socially desirable and/or normative influences in personality assessment. Targets rated their own personalities and recommended another person to also do so along shared sets of items diversely contaminated with socially desirable content. The recommended informant then invited a third individual to additionally make ratings of the original target. Profile correlations, analysis of variances (ANOVAs), and simple patterns of agreement/disagreement consistently converged on a strong normative effect paralleling item desirability, with all three rater types exhibiting a tendency to reject socially undesirable descriptors while also endorsing desirable indicators. These tendencies were, in fact, more prominent for informants than they were for self-raters. In their entirety, our results provide a note of caution regarding the strategy of using non-self informants as a comforting comparative benchmark within psychological measurement applications.

Exploitation of dihydroorotate dehydrogenase (DHODH) and p53 activation as therapeutic targets: A case study in polypharmacology

The tenovins are a frequently studied class of compounds capable of inhibiting sirtuin activity, which is thought to result in increased acetylation and protection of the tumor suppressor p53 from degradation. However, as we and other laboratories have shown previously, certain tenovins are also capable of inhibiting autophagic flux, demonstrating the ability of these compounds to engage with more than one target. In this study, we present two additional mechanisms by which tenovins are able to activate p53 and kill tumor cells in culture. These mechanisms are the inhibition of a key enzyme of the de novo pyrimidine synthesis pathway, dihydroorotate dehydrogenase (DHODH), and the blockage of uridine transport into cells. These findings hold a 3-fold significance: first, we demonstrate that tenovins, and perhaps other compounds that activate p53, may activate p53 by more than one mechanism; second, that work previously conducted with certain tenovins as SirT1 inhibitors should additionally be viewed through the lens of DHODH inhibition as this is a major contributor to the mechanism of action of the most widely used tenovins; and finally, that small changes in the structure of a small molecule can lead to a dramatic change in the target profile of the molecule even when the phenotypic readout remains static.