Camostat mesylate inhibits SARS-CoV-2 activation by TMPRSS2-related proteases and its metabolite GBPA exerts antiviral activity.

BACKGROUND: Antivirals are needed to combat the COVID-19 pandemic, which is caused by SARS-CoV-2. The clinically-proven protease inhibitor Camostat mesylate inhibits SARS-CoV-2 infection by blocking the virus-activating host cell protease TMPRSS2. However, antiviral activity of Camostat mesylate metabolites and potential viral resistance have not been analyzed. Moreover, antiviral activity of Camostat mesylate in human lung tissue remains to be demonstrated. METHODS: We used recombinant TMPRSS2, reporter particles bearing the spike protein of SARS-CoV-2 or authentic SARS-CoV-2 to assess inhibition of TMPRSS2 and viral entry, respectively, by Camostat mesylate and its metabolite GBPA. FINDINGS: We show that several TMPRSS2-related proteases activate SARS-CoV-2 and that two, TMPRSS11D and TMPRSS13, are robustly expressed in the upper respiratory tract. However, entry mediated by these proteases was blocked by Camostat mesylate. The Camostat metabolite GBPA inhibited recombinant TMPRSS2 with reduced efficiency as compared to Camostat mesylate. In contrast, both inhibitors exhibited similar antiviral activity and this correlated with the rapid conversion of Camostat mesylate into GBPA in the presence of serum. Finally, Camostat mesylate and GBPA blocked SARS-CoV-2 spread in human lung tissue ex vivo and the related protease inhibitor Nafamostat mesylate exerted augmented antiviral activity. INTERPRETATION: Our results suggest that SARS-CoV-2 can use TMPRSS2 and closely related proteases for spread in the upper respiratory tract and that spread in the human lung can be blocked by Camostat mesylate and its metabolite GBPA. FUNDING: NIH, Damon Runyon Foundation, ACS, NYCT, DFG, EU, Berlin Mathematics center MATH+, BMBF, Lower Saxony, Lundbeck Foundation, Novo Nordisk Foundation

Behavioural Phenotyping of APPswe/PS1δE9 Mice: Age-Rrelated Changes and Effect of Long-Term Paroxetine Treatment.

Alzheimer's disease (AD) is a devastating illness characterized by a progressive loss of cognitive, social, and emotional functions, including memory impairments and more global cognitive deficits. Clinical-epidemiological evidence suggests that neuropsychiatric symptoms precede the onset of cognitive symptoms both in humans with early and late onset AD. The behavioural profile promoted by the AD pathology is believed to associate with degeneration of the serotonergic system. Using the APPswe/PS1δE9 model of AD-like pathology starting with 9 months old mice, we characterised long term non-cognitive behavioural changes measured at 9, 12, 15, and 18 months of age and applied principal component analysis on data obtained from open field, elevated plus maze, and social interaction tests. Long-term treatment with the selective serotonin reuptake inhibitor (SSRI) paroxetine was applied to assess the role of 5-HT on the behavioural profile; duration of treatment was 9 months, initiated when mice were 9 months of age. Treatment with paroxetine delays the decline in locomotion, in exploration and risk assessment behaviour, found in the APP/PS1 mice. APP/PS1 mice also exhibit low social activity and less aggressiveness, both of which are not affected by treatment with paroxetine. The APP/PS1 behavioural phenotype, demonstrated in this study, only begins to manifest itself from 12 months of age. Our results indicate that treatment with SSRI might ameliorate some of the behavioural deficits found in aged APP/PS1 mice

A Placement for Everyone

‘A Placement for Everyone’ outlines an emergent project in which staff and students collaborate to envision a transformation in their university/art school. Rather than convey pre-existing knowledge and expertise from tutor to students, we are working together as equal but different participants in a reflexive experiment with the institutional conditions and social purpose of learning. A primary influence has been the Artist Placement Group (APG), which from 1966—1979 negotiated for artists to be placed in business, industry and governmental organizations. These placements were not framed as commissions or contracts between the artist and the host, but were developed around the APG concept of ‘the open brief’, with no prescriptions, and no outcomes agreed in advance. In December 2012, I designated my academic post at University of the Arts London (UAL) as an artist’s placement, with a remit to promote creative engagement with the ecological and social-economic crisis. I aimed to connect artistic research with pedagogy, by involving staff and students in aligning our university’s operations with its stated values

Abbreviation of behavioural variables scored during testing.

<p>Abbreviation of behavioural variables scored during testing.</p

Effect of age, genotype and treatment on performance in OF (open field) and EPM (elevated plus maze) tests, analysed by principal component analysis.

<p>Factor scores for 4 types of behaviour (principal components) are shown in A-D. MANOVA was used to analyse significance in effect of age, genotype and treatment, followed by Fisher’s LSD <i>post hoc</i> test for group-wise comparisons: *, **, ***—p < 0.05, 0.01, 0.001 correspond to differences in factor scores between groups at 9, 12, 15 or 18 months of age; <b>+, ++, +++—</b>p < 0.05, 0.01, and 0.001 correspond to age-effect within groups compared to factor scores at 9 months; ¤, ¤¤, ¤¤¤—p < 0.05, 0.01, 0.001 correspond to intra-group comparisons of factor scores at 12 months. Bars indicate Means ± SEM Black bars, Wt (n = 14–15); white bars, Tg (n = 12–13); dark grey bars, Wt-paroxetine treated (n = 14–16); light grey bars, Tg-paroxetine treated (n = 6–12, and n = 4 at 12 months of age). 9, 12, 15 and 18 months of age correspond to 0, 3, 6 and 9 months of treatment, respectively.</p

The serum paroxetine levels assessed by UHPLC-MS/MS.

<p>The serum paroxetine levels assessed by UHPLC-MS/MS.</p

Effect of age, genotype and treatment on social interaction, tested on two consecutive days, analysed by principal component analysis.

<p>Factor scores for 4 types of behaviour (principal components) are shown in A-D. MANOVA was used to analyse significance in effect of age, genotype and treatment, following by Fisher’s LSD <i>post hoc</i> test for group-wise comparisons: *, **, ***—p < 0.05, 0.01, 0.001 correspond to differences in factor scores between groups at 9, 12, 15 or 18 months of age; +, ++, +++—p < 0.05, 0.01, and 0.001 correspond to age-effect within groups compared to factor scores at 9 months; ¤, ¤¤, ¤¤¤—p < 0.05, 0.01, 0.001 correspond to intra-group comparisons of factor scores at 12 months. Bars indicate Means ± SEM Black bars, Wt (n = 15–23); white bars, Tg (n = 12–22); dark grey bars, Wt-paroxetine treated (n = 14–16); light grey bars, Tg-paroxetine treated (n = 7–12, and n = 3 at 12 months of age). 9, 12, 15 and 18 months of age correspond to 0, 3, 6 and 9 months of treatment, respectively.</p

Pharmacokinetics and pharmacodynamics of cannabis‐based medicine in a patient population included in a randomized, placebo‐controlled, clinical trial

Abstract Information on the pharmacokinetics (PK) and pharmacodynamics (PD) of orally administered cannabis‐based medicine (CBM) in capsule formulation in patient populations is sparse. In this exploratory study, we aimed to evaluate the PK and PD in a probable steady state of CBM in neuropathic pain and spasticity in a population of patients with multiple sclerosis (MS). Of 134 patients participating in a randomized, double‐blinded, placebo‐controlled, trial, 23 patients with MS (17 female) mean age 52 years (range 21–67) were enrolled in this substudy. They received oral capsules containing Δ9‐tetrahydrocannabinol (THC, n = 4), cannabidiol (CBD, n = 6), a combination (THC&CBD, n = 4), or placebo (n = 9). Maximum doses were 22.5 mg (THC) and 45 mg (CBD) a day divided into three administrations. PD parameters were evaluated for pain and spasticity. Blood samples were analyzed using an ultra‐high‐performance liquid chromatography–tandem mass spectrometer after protein precipitation and phospholipid removal. PK parameters were estimated using computerized modeling. The variation in daily dose and PK between individuals was considerable in a steady state, yet comparable with previous reports from healthy controls. Based on a simulation of the best model, the estimated PK parameters (mean) for THC (5 mg) were Cmax 1.21 ng/mL, Tmax 2.68 h, and half‐life 2.75 h, and for CBD (10 mg) were Cmax 2.67 ng/mL, Tmax 0.10 h, and half‐life 4.95 h, respectively. No effect was found on the PD parameters, but the placebo response was considerable. More immediate adverse events were registered in the active treatment groups compared with the placebo group