Time-dependent biphasic modulation of human BDNF by antidepressants in neuroblastoma cells

<p>Abstract</p> <p>Background</p> <p>Recent rodent studies reported that antidepressant treatments affect the expression of brain-derived neurotrophic factor (BDNF) mRNA in a way that is dependent on treatment duration, by selective modulation of different BDNF transcripts. However, no data are available for the human BDNF gene. We studied the effect of different antidepressants on BDNF mRNA expression in human neuroblastoma SH-SY5Y cells.</p> <p>Results</p> <p>Cultured cells were treated with the antidepressants fluoxetine, reboxetine and desipramine for different time lengths (6, 24, 48 hours). Expression of total BDNF mRNA was analyzed by reverse transcription PCR and levels of different BDNF transcripts were detected by hemi-nested PCR with specific primers.</p> <p>Short-term treatment (6 hours) with reboxetine or desipramine reduced total BDNF, whereas long-term treatment (48 hours) significantly increased total BDNF mRNA levels. These changes were accounted for by differential regulation of BDNF IV and VIa/b transcripts. Fluoxetine showed no significant effects.</p> <p>Conclusion</p> <p>This is the first study showing biphasic changes in the expression of total and specific BDNF transcripts in human cells following antidepressant treatments. These findings suggest that biphasic induction of BDNF by antidepressants could be a feature common to rodents and humans and encourage the use of SH-SY5Y cells as a tool for investigation of drug effects on human genes.</p

Nirmatrelvir treatment of SARS-CoV-2-infected mice blunts antiviral adaptive immune responses

Alongside vaccines, antiviral drugs are becoming an integral part of our response to the SARS-CoV-2 pandemic. Nirmatrelvir-an orally available inhibitor of the 3-chymotrypsin-like cysteine protease-has been shown to reduce the risk of progression to severe COVID-19. However, the impact of nirmatrelvir treatment on the development of SARS-CoV-2-specific adaptive immune responses is unknown. Here, by using mouse models of SARS-CoV-2 infection, we show that nirmatrelvir administration blunts the development of SARS-CoV-2-specific antibody and T cell responses. Accordingly, upon secondary challenge, nirmatrelvir-treated mice recruited significantly fewer memory T and B cells to the infected lungs and mediastinal lymph nodes, respectively. Together, the data highlight a potential negative impact of nirmatrelvir treatment with important implications for clinical management and might help explain the virological and/or symptomatic relapse after treatment completion reported in some individuals

Preparation of arylaminocarbonyl bicyclic sulfonamides as inhibitors of hepatitis B virus

The invention relates to prepn. of arylaminocarbonyl bicyclic sulfonamides (I) that are inhibitors of hepatitis B virus (HBV)​. Compds. I wherein A is a 6-​membered arom. or heteroarom. ring; B is a 6-​membered aryl optionally contg. one or more N atoms; X is H or NR3R4; Y is a H, halo, NH2, etc.; R1 and R2 each independently is H, linear or branched C1-​6 alkyl, etc.; R3 and R4 each independently is H, linear or branched C1-​3 alkyl, etc.; etc., are claimed. The example compd. II was prepd. from an intermediate(also prepd.) and methanamine (procedure given)​. The compds. of the invention were evaluated for their biol. activity (data given)​. Compds. I are useful alone or in combination with other agents for treating, ameliorating, preventing or curing HBV infection and related conditions. The invention also relates to pharmaceutical compns. contg. I. The invention relates to prepn. of arylaminocarbonyl bicyclic sulfonamides (I) that are inhibitors of hepatitis B virus (HBV)​. Compds. I wherein A is a 6-​membered arom. or heteroarom. ring; B is a 6-​membered aryl optionally contg. one or more N atoms; X is H or NR3R4; Y is a H, halo, NH2, etc.; R1 and R2 each independently is H, linear or branched C1-​6 alkyl, etc.; R3 and R4 is H, each independently is H, linear or branched C1-​3 alkyl,​. The example compd. II was prepd. from an intermediate(also prepd.) and methanamine (procedure given)​. The compds. of the invention were evaluated for their biol. activity (data given)​. Compds. I are useful alone or in combination with other agents for treating, ameliorating, preventing or curing HBV infection and related conditions. The invention also relates to pharmaceutical compns. contg. I

Preparation of arylaminocarbonyl bicyclic sulfonamides as inhibitors of hepatitis B virus

The invention relates to prepn. of arylaminocarbonyl bicyclic sulfonamides (I) that are inhibitors of hepatitis B virus (HBV)​. Compds. I wherein B is a 6-​membered aryl optionally contg. one or more N atoms; X is N (un)​substituted with R4, or is O; Y is a single bond, a double bond, C1-​4 alkanediyl, etc.; Z is C(O) or CR2R3; R1 is H, OH, etc.; R2 is H, DNH2, etc.; R3 is absent, H, D, etc.; R4 is H, OH, C1-​4 alkyl; R5 is C1-​3 alkyl (un)​substituted with OH; R6 is C1-​4 alkyl; etc., are claimed. The example compd. II was prepd. from an intermediate (also prepd.) and formaldehyde (procedure given)​. The compds. of the invention were evaluated for their biol. activity (data given)​. Compds. I are useful alone or in combination with other agents for treating, ameliorating, preventing or curing HBV infection and related conditions. The invention also relates to pharmaceutical compns. contg. I

Preparation of oxalamido-​substituted tricyclic inhibitors of hepatitis B virus

The present invention relates to compds. I [Cy = aryl or heteroaryl; A = CR3 or N; X = O, S, NH, SO, SO2 or a single bond; Y, Y1-​Y3 = (independently) (un)​substituted alkanediyl or alkenediyl, or a single bond; R1 = H or alkyl; R2 = H, OH, halo and alkyl; R3 = H, alkyl, cycloalkyl, haloalkyl, halo; R5 = H or alkyl; or R2 and R5 taken together form alkanediyl bridge; R6 = H, alkyl, cycloalkyl, heterocycloalkyl, etc.; each of R7 and R8 = (independently) H, (un)​substituted alkyl, aryl, heteroaryl, etc.; or R7 and R8 form together with the nitrogen atom to which they are attached a cyclic amine selected from aziridine, azetidine, pyrrolidine, etc.; R, Ra, Rb, Rc = (independently) H, halo, CN, alkyl, etc.] or pharmaceutically acceptable salts, tautomers, solvates or stereoisomers thereof, that are inhibitors of hepatitis B virus (HBV)​. For example, reacting cis-​7-​methyl-​N-​(3,​4,​5-​trifluorophenyl)​-​2,​3,​3a,​4,​10,​10a-​hexahydro-​1H,​7H-​dipyrrolo[3,​4-​b:3',​4'-​f]​[1,​4,​5]​oxathiazocine-​8-​carboxamide 5,​5-​dioxide hydroiodide with N,​N-​dimethyloxamic acid afforded cis-​II. The exemplified compds. I were tested for HBV inhibition (data given)​. Compds. I are useful alone or in combination with other agents for treating, ameliorating, preventing or curing HBV infection and related conditions. The present invention also relates to pharmaceutical compns. contg. said compds. I

Reversibility of HCV-induced changes in PI4P subcellular distribution.

<p>JFH-A4 cells were incubated for 14 days with the HCV RdRP inhibitor HCV-796 (2 µM) or the HCV NS3/4A protease inhibitor MK-5172 (0.2 µM). Cure from HCV was controlled by detection of NS5A with a specific NS5A antibody (red, right column). As control, untreated Huh7.5 cells or JFH-A4 cells were used. Cells were fixed and PI4P (green) was detected in the internal membranes (IM, left column) or in the plasma membrane (PM, central column). For internal membrane staining giantin (red) was used as a specific marker for Golgi membranes. Nuclei were stained by the Hoechst dye (blue).</p

HCV impacts subcellular PI4P distribution.

<p>(A) Huh7.5 cells, JFH-A4 and Con1-SR cells were analyzed by confocal microscopy for the presence of PI4P (green) in the plasma membranes (PM, upper panel) or in the intracellular membrane (IM, lower panel) using the protocols described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002576#s4" target="_blank">Materials and Methods</a>. Nuclei were stained by the Hoechst dye (blue). For internal membrane staining, giantin (red) was used as a specific marker for Golgi membranes. (B) Quantification of PI4P levels by immunofluorescence analysis. Changes in mean fluorescence intensity relative to the control (Huh7.5 cells) are shown. Four randomly picked fields were analyzed per each condition. Normalization was performed as detailed in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002576#s4" target="_blank">Materials and Methods</a>. Data are presented as averages ± SEM. *, p<0.05; **, p<0.01; ***, p<0.001.</p