9 research outputs found
StructureâActivity Relationship Study around Guanabenz Identifies Two Derivatives Retaining Antiprion Activity but Having Lost α2-Adrenergic Receptor Agonistic Activity
Guanabenz
(GA) is an orally active α2-adrenergic agonist
that has been used for many years for the treatment of hypertension.
We recently described that GA is also active against both yeast and
mammalian prions in an α2-adrenergic receptor-independent manner.
These data suggest that this side-activity of GA could be explored
for the treatment of prion-based diseases and other amyloid-based
disorders. In this perspective, the potent antihypertensive activity
of GA happens to be an annoying side-effect that could limit its use.
In order to get rid of GA agonist activity at α2-adrenergic
receptors, we performed a structureâactivity relationship study
around GA based on changes of the chlorine positions on the benzene
moiety and then on the modifications of the guanidine group. Hence,
we identified the two derivatives <b>6</b> and <b>7</b> that still possess a potent antiprion activity but were totally
devoid of any agonist activity at α2-adrenergic receptors. Similarly
to GA, <b>6</b> and <b>7</b> were also able to inhibit
the protein folding activity of the ribosome (PFAR) which has been
suggested to be involved in prion appearance/maintenance. Therefore,
these two GA derivatives are worth being considered as drug candidates
Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma
<div><p>Background</p><p>Allergic asthma is caused by abnormal immunoreactivity against allergens such as house dust mites among which <i>Dermatophagoides farinae</i> (Der f) is a common species. Currently, immunotherapy is based on allergen administration, which has variable effect from patient to patient and may cause serious side effects, principally the sustained risk of anaphylaxis. DNA vaccination is a promising approach by triggering a specific immune response with reduced allergenicity.</p><p>Objective</p><p>The aim of the study is to evaluate the effects of DNA immunization with Der f1 allergen specific DNA on allergic sensitization, inflammation and respiratory function in mice.</p><p>Methods</p><p>Mice were vaccinated 28 and 7 days before allergen exposure with a Der f1-encoding plasmid formulated with a block copolymer. Asthma was induced by skin sensitization followed by intra-nasal challenges with Der f extract. Total lung, broncho-alveolar lavage (BAL) and spleen cells were analyzed by flow cytometry for their surface antigen and cytokine expression. Splenocytes and lung cell IFN-Îł production by CD8+ cells in response to Der f CMH1-restricted peptides was assessed by ELISPOT. IgE, IgG1 and IgG2a were measured in serum by ELISA. Specific bronchial hyperresponsiveness was assessed by direct resistance measurements.</p><p>Results</p><p>Compared to animals vaccinated with an irrelevant plasmid, pVAX-Der f1 vaccination induced an increase of B cells in BAL, and an elevation of IL-10 and IFN-Îł but also of IL-4, IL-13 and IL-17 producing CD4+ lymphocytes in lungs and of IL-4 and IL-5 in spleen. In response to CD8-restricted peptides an increase of IFN-Îł was observed among lung cells. IgG2a levels non-specifically increased following block copolymer/DNA vaccination although IgE, IgG1 levels and airways resistances were not impacted.</p><p>Conclusions & Clinical Relevance</p><p>DNA vaccination using a plasmid coding for Der f1 formulated with the block copolymer 704 induces a specific immune response in the model of asthma used herein.</p></div
Der f1 and ÎČgal expression assessment.
<p>Five days after the last vaccine injection, Der f1 and ÎČgal gene expression were verified by RT-PCR on skeletal muscle RNA using specific primers (nâ=â3 per group). This experience was done for both DNA vaccination conditions (10 ”g and 5 ”g of plasmid preparation).</p
Immunization protocols against Der f1 in asthmatic mice.
<p>pVAX-Der f1 or pCMV-ÎČgal plasmid were injected i.m at days â28 and â7. Mice were then epicutaneously sensitized and intranasally challenged with total extract of HDM. Analyses were performed on day 35.</p
Effect of immunization protocol on BAL inflammation of non asthmatic non vaccinated mice (light grey bar, nâ=â8), Der f mice (open bar, nâ=â9), pCMV-ÎČgal (grey bar, nâ=â7) and pVAX-Der f1 mice (black bar, nâ=â7) when using 10 ”g of DNA.
<p>The total number of cells was determined by cell count on Kova slides. The cellular composition was established by flow cytometry. Results are expressed as absolute number of cells, as the mean and standard deviation for each group. **p<0,01 and *** p<0,001 by Mann Whitney test.</p
Effect of prophylactic immunization protocol with 10 ”g of Der f1 DNA on respiratory function.
<p>Airway resistance and Compliance was measured at day 35 using Flexivent with instillation of 5 to 20/ml methacholine in non asthmatic non vaccinated mice (nâ=â6, ) Der f (nâ=â7, O ), pCMV-ÎČgal mice (nâ=â9,) and pVAX-Der f1 (nâ=â9,âŸ) mice. Results are expressed in increased fold, as a mean for each group ± standard deviation. *p<0,05 by Mann Whitney test.</p
Flow cytometry mix for detection of cytokine produced by lung T cells.
<p>Flow cytometry mix for detection of cytokine produced by lung T cells.</p
Effect of immunization protocol on T cells cytokines secretion of Der f mice (open bar, nâ=â6), pCMV-ÎČgal (grey bar, nâ=â7) and pVAX-Der f1 (black bar, nâ=â7) when using 10 ”g of DNA.
<p>One day after the last airway allergen challenge, mice were sacrificed and lung cells were cultured. The concentration of IL-4, IL-5, IL-13, IFN-Îł, IL-10 and IL-17 were measured by flow cytometry. Results are expressed as the mean and standard deviation for each group. *p<0,05, ** p<0,01 and *** p<0,001 by Mann Whitney test.</p
Cytokine concentrations (pg/ml) in vaccinated mice spleen cell culture supernatants after stimulation by medium of Der f1.
*<p>p<0,05 between pCMV-ÎČgal and pVAX-Der f1 and <sup>$</sup> p<0,05 between Der f and pVAX-Der f1.</p