Design, synthesis and biological evaluation of piperazino-enaminones as novel suppressants of pro-Inflammatory cytokines.

Infection triggers the release of pro-inflammatory cytokines (TNF-alpha and IL-6). Over-production, however, cause tissue injury seen in severe asthma. The ability of enaminone E121 to reduce pro-inflammatory cytokines in our laboratory encouraged further examination of its structural scaffold. Piperazino-enaminones were designed by incorporating n-arylpiperazine motif into the aromatic enaminone. Four possible modifications were explored systematically. Synthesis was accomplished by amination of the corresponding methyl/ethyl 2,4-dioxo-6-(substituted)cyclohexane-carboxylate.. Sixteen novel compounds were synthesized. Biological activity was tested in J774 macrophages stimulated with lipopolysaccharides. The release of cytokines was measured via ELISA. Four compounds significantly suppressed TNF-alpha and IL-6 release in dose-dependent manner.

A questionnaire based survey among pharmacy practitioners to evaluate the level of knowledge and confidence towards antimicrobial stewardship.

Objective: Our study aimed to assess the knowledge, understanding and confidence of the practicing pharmacists in UAE as an antimicrobial Stewards. Antimicrobial resistance threatens the achievements of modern medicine globally, and it's highly required for the AMS principles to be implemented in our communities. Methods: A cross-sectional online- questionnaire based survey was used among UAE pharmacy practitioners from different areas of practice who are holding pharmaceutical degrees and/or licensed pharmacists. The questionnaire was sent to the participants via social media platforms. The questionnaire was validated, and reliability assessment was made prior to the conduct. Results: A total of 117 pharmacists responded to this study, out of which (70.9%, n=83) were females. Pharmacists which are from various practice fields participated in the survey, but the majority were pharmacists in Hospital pharmacies or Clinical pharmacists (47%, n=55), also community pharmacists (35.9%, n=42), while only (16.9%, n=20) ware from other areas of pharmacy including industrial pharmacy and academia. The majority of participants 88.9% (n= 104) were interested in pursuing their career as an Infectious disease pharmacist or getting a certificate in antimicrobial stewardship. The mean scores in the knowledge towards antimicrobial resistance was 3.75 (poor: 1-1.6, moderate: 1.7-3.3. Good: 3.4-5), indicates that the pharmacists have a good level of knowledge towards AMR. A total of 84.3% of participants succeeded in Identifying the correct intervention for antibiotic resistance. The findings also showed that the total mean score of hospital pharmacists (mean=10.6±1.12), and the average of the scores of community pharmacists (mean=9.8±1.38), were non-significant between the different area of practice. 52.3% of the participants had a training on antimicrobial stewardship during their experiential rotation which reflected on their confidence in their performance and knowledge assessment (p value < 0.05). Conclusion: The study concluded good knowledge and high confidence levels among practicing pharmacists in UAE. However, the findings also identify areas of improvement in the practicing pharmacist, and the significant relationship between the knowledge and confidence scores reflects the ability of the practicing pharmacists to integrate the AMS principles within the UAE, which aligns with the attainability of the improvement.

A questionnaire based survey among pharmacy practitioners to evaluate the level of knowledge and confidence towards antimicrobial stewardship.

Objective: Our study aimed to assess the knowledge, understanding and confidence of the practicing pharmacists in UAE as an antimicrobial Stewards. Antimicrobial resistance threatens the achievements of modern medicine globally, and it's highly required for the AMS principles to be implemented in our communities. Methods: A cross-sectional online- questionnaire based survey was used among UAE pharmacy practitioners from different areas of practice who are holding pharmaceutical degrees and/or licensed pharmacists. The questionnaire was sent to the participants via social media platforms. The questionnaire was validated, and reliability assessment was made prior to the conduct. Results: A total of 117 pharmacists responded to this study, out of which (70.9%, n=83) were females. Pharmacists which are from various practice fields participated in the survey, but the majority were pharmacists in Hospital pharmacies or Clinical pharmacists (47%, n=55), also community pharmacists (35.9%, n=42), while only (16.9%, n=20) ware from other areas of pharmacy including industrial pharmacy and academia. The majority of participants 88.9% (n= 104) were interested in pursuing their career as an Infectious disease pharmacist or getting a certificate in antimicrobial stewardship. The mean scores in the knowledge towards antimicrobial resistance was 3.75 (poor: 1-1.6, moderate: 1.7-3.3, Good: 3.4-5), indicates that the pharmacists have a good level of knowledge towards AMR. A total of 84.3% of participants succeeded in Identifying the correct intervention for antibiotic resistance. The findings also showed that the total mean score of hospital pharmacists (mean=10.6±1.12), and the average of the scores of community pharmacists (mean=9.8±1.38), were non-significant between the different area of practice. 52.3% of the participants had a training on antimicrobial stewardship during their experiential rotation which reflected on their confidence in their performance and knowledge assessment (p value < 0.05). Conclusion: The study concluded good knowledge and high confidence levels among practicing pharmacists in UAE. However, the findings also identify areas of improvement in the practicing pharmacist, and the significant relationship between the knowledge and confidence scores reflects the ability of the practicing pharmacists to integrate the AMS principles within the UAE, which aligns with the attainability of the improvement.

The novel piperazino-enaminone JOAB-40 reduced colitis severity in mice via inhibition tumor necrosis factor-α and interleukin-1ß.

BRIEF INTRODUCTION: The synthetic compound enaminone E121 has an established role as a potent anti-tussive, bronchodilator and anti-inflammatory agent in asthma, cough, and colitis induced animal models. The addition of an N-alkylated piperazine motif to the terminal end of E121 lead to the generation of various analogues such as JOAB-40. JOAB-40 was shown to be more potent than the lead compound E121 in inhibiting the expression of the chemokine receptor CCR2, ERK1/2 phosphorylation, and the release of pro-inflammatory cytokines in vitro. MAIN OBJECTIVE OF THE STUDY: We hypothesize that JOAB-40 is more potent than the lead compound E121 in reducing colitis severity in mice in part through inhibiting the release of TNFα and IL-1ß. METHODS: Colitis was induced by dextran sulfate sodium (DSS) administration using prophylactic and treatment approaches. The severity of the inflammation was determined by the gross (macroscopic) and histological (microscopic) assessments. The levels of TNFα, IL-1ß, and IL-10 release in response to lipopolysaccharide (LPS) stimulation from the adherent murine macrophage cell line J774.2 in vitro, and the circulating levels of TNFα in vivo was measured by ELISA-based technique. SIGNIFICANT FINDINGS FROM THE STUDY: E121 administration (1-60 mg/kg) in mice with established colitis (treatment approach) did not reduce colitis severity. On the other hand, JOAB-40 administration significantly reduced colitis severity in mice when administered using two approaches; a) prophylactic (given along colitis induction), and b) treatment (given after colitis was established) with doses as low as 10 mg/kg. The degree of inhibition of TNFα and IL-1ß (but not IL-10) release from J774.2 cell line in response to LPS stimulation was more potent with JOAB-40 than E121. This was also observed in vivo in regards to the circulating levels of TNFα. RELEVANT CONTRIBUTION TO KNOWLEDGE: Our results indicate that JOAB-40 is more potent than E121 in reducing colitis severity in mice and may be a promising future therapeutic target for the management of inflammatory bowel disease (IBD).

A questionnaire based survey among pharmacy practitioners to evaluate the level of knowledge and confidence towards antimicrobial stewardship

Objective: Our study aimed to assess the knowledge, understanding and confidence of the practicing pharmacists in UAE as an antimicrobial Stewards. Antimicrobial resistance threatens the achievements of modern medicine globally, and it’s highly required for the AMS principles to be implemented in our communities. Methods: A cross-sectional online- questionnaire based survey was used among UAE pharmacy practitioners from different areas of practice who are holding pharmaceutical degrees and/or licensed pharmacists. The questionnaire was sent to the participants via social media platforms. The questionnaire was validated, and reliability assessment was made prior to the conduct. Results: A total of 117 pharmacists responded to this study, out of which (70.9%, n=83) were females. Pharmacists which are from various practice fields participated in the survey, but the majority were pharmacists in Hospital pharmacies or Clinical pharmacists (47%, n=55), also community pharmacists (35.9%, n=42), while only (16.9%, n=20) ware from other areas of pharmacy including industrial pharmacy and academia. The majority of participants 88.9% (n= 104) were interested in pursuing their career as an Infectious disease pharmacist or getting a certificate in antimicrobial stewardship. The mean scores in the knowledge towards antimicrobial resistance was 3.75 (poor: 1-1.6, moderate: 1.7-3.3, Good: 3.4-5), indicates that the pharmacists have a good level of knowledge towards AMR. A total of 84.3% of participants succeeded in Identifying the correct intervention for antibiotic resistance. The findings also showed that the total mean score of hospital pharmacists (mean=10.6±1.12), and the average of the scores of community pharmacists (mean=9.8±1.38), were non-significant between the different area of practice (AU)

A Retrospective Study of the Cytokine Profile Changes in Mice with FVIII Inhibitor Development After Adeno-Associated Virus-Mediated Gene Therapy in a Hemophilia A Mouse Model.

The development of inhibitory autoantibodies to the infused clotting factor VIII (FVIII) is a major complication for severe hemophilia A management. Novel therapy options for hemophilia have significantly progressed in the last decade, and a gene therapy cure for hemophilia is becoming a reality. However, mechanistic studies of FVIII autoantibodies (FVIII inhibitors) have lagged behind and remain a challenge for both protein replacement and gene therapy. FVIII inhibitor formation is assumed to be a classical T cell-dependent immune response in which cytokines/chemokines play an important role. The study of cytokine profile changes during FVIII inhibitor development may be helpful to understand the mechanism of inhibitor development and to explore potential novel approaches that will minimize the risk. After FVIII-/- mice were treated with intravenous administration of an adeno-associated virus 8 vector encoding human FVIII, FVIII expression peaked at week 2 (W2), and FVIII inhibitor was thoroughly developed at week 8 (W8). W8 plasma that showed positive FVIII inhibitor, and W2 samples with negative FVIII inhibitor (anti-FVIII[+]), were subjected to multiplex cytokines measurement. W8 and W2 samples were both negative for FVIII inhibitor (anti-FVIII[-]) as the control. In comparison to mice in the anti-FVIII(-) group, mice in the anti-FVIII(+) group exhibited significantly elevated pro-inflammatory cytokines of interleukin (IL)-1, IL-6, IL-12p40, monocyte chemoattractant protein-1, macrophage inflammatory protein (MIP)-1, MIP-2, and tumor necrosis factor alpha (TNF-α), especially at higher titers. The anti-inflammatory cytokine of transforming growth factor beta (TGF-ß) was decreased at W2 in both groups. Multivariate analysis of the risk factors for FVIII inhibitor development showed peak FVIII activity at W2. IL-6 and TNF-α at W8 were positively correlated with inhibitor formation, and negatively correlated with the age starting gene therapy. Collectively, the elevated monocyte derived pro-inflammatory cytokines/chemokines, together with the decreased anti-inflammatory cytokine of TGF-ß at an early time point, may contribute to the persistent inflammatory environment in favor of an immune response toward FVIII inhibitor development.

Current and novel anti-inflammatory drug targets for inhibition of cytokines and leucocyte recruitment in rheumatic diseases.

OBJECTIVES: Many studies of disease state mechanisms reveal that unbridled inflammation is to blame for many of the symptoms associated with autoimmune diseases such as Crohn's and Rheumatoid Arthritis (RA). While therapies aimed at decreasing levels of pro-inflammatory cytokines exist, some have failed clinically or have extensive adverse effects. The aim of this review is to discuss common drug targets for anti-inflammatory therapies as well as explore potential mechanisms of action for new therapies. Various studies done on novel mechanisms targeting pro-inflammatory cytokine release as well as leukocyte chemotaxis have been researched for discussion here. Both of these contribute to tissue injury and patient symptoms in inflammatory and autoimmune disease states. KEY FINDINGS: While many current drug targets suppress inflammation via the receptor, research aimed at identifying new compounds and signaling mechanisms is ongoing to identify new targets within pro-inflammatory signaling pathways, or specific immune cell types. CONCLUSIONS: While glucocorticoids and monoclonal antibodies have shown to be efficacious, some patients have encountered mixed results. Biologic therapies also come with a high price tag Thus, novel compounds with new immune drug targets are ideal for patients whose therapies have not been successful.

A questionnaire based survey among pharmacy practitioners to evaluate the level of knowledge and confidence towards antimicrobial stewardship

Objective: Our study aimed to assess the knowledge, understanding and confidence of the practicing pharmacists in UAE as an antimicrobial Stewards. Antimicrobial resistance threatens the achievements of modern medicine globally, and it’s highly required for the AMS principles to be implemented in our communities. Methods: A cross-sectional online- questionnaire based survey was used among UAE pharmacy practitioners from different areas of practice who are holding pharmaceutical degrees and/or licensed pharmacists. The questionnaire was sent to the participants via social media platforms. The questionnaire was validated, and reliability assessment was made prior to the conduct. Results: A total of 117 pharmacists responded to this study, out of which (70.9%, n=83) were females. Pharmacists which are from various practice fields participated in the survey, but the majority were pharmacists in Hospital pharmacies or Clinical pharmacists (47%, n=55), also community pharmacists (35.9%, n=42), while only (16.9%, n=20) ware from other areas of pharmacy including industrial pharmacy and academia. The majority of participants 88.9% (n= 104) were interested in pursuing their career as an Infectious disease pharmacist or getting a certificate in antimicrobial stewardship. The mean scores in the knowledge towards antimicrobial resistance was 3.75 (poor: 1-1.6, moderate: 1.7-3.3, Good: 3.4-5), indicates that the pharmacists have a good level of knowledge towards AMR. A total of 84.3% of participants succeeded in Identifying the correct intervention for antibiotic resistance. The findings also showed that the total mean score of hospital pharmacists (mean=10.6±1.12), and the average of the scores of community pharmacists (mean=9.8±1.38), were non-significant between the different area of practice.(AU)

Novel Piperazino-Enaminones Suppress Pro-Inflammatory Cytokines and Inhibit Chemokine Receptor CCR2.

Pro-inflammatory mediators including TNF-alpha, IL-6, and nitric oxide are important for the regulation of the immune response when an infection is present, but when overproduced, it can be responsible for the development of tissue and organ injury seen in sepsis, as well as severe asthma, and autoimmune diseases such as Crohn's disease and rheumatoid arthritis. Data from our lab to characterize the novel compound enaminone E121 have suggested that macrophages stimulated with lipopolysaccharide (LPS) release significantly decreased levels of TNF-alpha and IL-6 as measured by enzyme-linked immunosorbent assay as compared to the DMSO control group. Additionally, functional experiments in a mouse model of asthma have shown that E121 is efficacious in decreasing airway hyperresponsiveness. A new set of compounds synthesized in our lab (JODI) have an N-aryl piperazino motif incorporated on the aromatic side of the enaminone pharmacophore. It was hypothesized that this would enhance their immunosuppressive activity as anti-inflammatory agents by also acting as a chemokine receptor antagonist. Our studies suggest that JODI appears to suppress TNF-alpha and IL-6 in a dose-dependent manner. The JODI compounds were also more effective in reducing TNF-alpha after LPS stimulation when compared to dexamethasone. Lastly, studies using MCP-1 suggest that the JODI compounds, and not E121, are able to block CCR2 signaling as evidenced by decreased total ERK1/2. These studies indicate that E121 and its corresponding piperazino analogs could act as strong anti-inflammatory agents in asthma or other autoimmunities where efficacious therapeutic options are needed.