A Sonochemical Access to 5-Aryl Substituted Pyrazolo[1,5-a]Pyrimidines as Potential Inhibitors of TNF-α

In spite of various and extensive studies known for pyrazolo[1,5-a]pyrimidines the synthesis, in silico studies and biological evaluation of their 5-(het)aryl analogs remained underexplored. The TNF-α inhibitors on the other hand has considerable therapeutic potential for autoimmune and inflammatory diseases in addition to cancer, diabetes and possibly COVID-19. In the current study 5-aryl pyrazolo[1,5-a]pyrimidines were explored as potential inhibitors of TNF-α that was supported by the in silico studies. This class of compounds was accessed via a sonochemical synthesis involving the acid catalyzed cyclocondensation reaction of aminopyrazoles with acrylophenones in the presence of aerial oxygen. The study indicated that the overall rate of the reaction was enhanced by ultrasound in the absence of which a longer duration and higher temperature was necessary. The current catalyst/promoter/ligand free and scalable method afforded a range of compounds. Some of these compounds showed good inhibition of TNF-α in vitro where ester/amide moiety at the C-3 position played a key role in interacting with the protein dimer as suggested by the in silico studies. Indeed, these groups formed H-bonds with A: GLY121 and B: TYR151 residues of TNF-α dimer in silico. A brief SAR within the series and in silico ADME/toxicity prediction for best active compounds is presented. Compounds 3a-c were identified as initial hits for further pharmacological evaluations. A sonochemical method has been developed for the facile synthesis of pyrazolo[1,5-a]pyrimidines that were evaluated as potential inhibitors of TNF-α. [ABSTRACT FROM AUTHOR] Copyright of Polycyclic Aromatic Compounds is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

An In-vitro evaluation of a polyherbal formulation, against SARS-Cov-2.

Background: In the last two years, COVID-19 pandemic caused by SARS-CoV-2 has created a mass destruction among humanity causing a major health crisis around the world. With the emergence of new strains of the virus, lack of targeted drugs and antimicrobial resistance, there is a dire need to discover specific antiviral with minimum side effects targeted against COVID-19. Objective: The present study evaluates the antiviral efficacy of a novel Ayurvedic polyherbal formulation, NOQ19, composed of a 13 well known herbs, in a cell-based setting. Methodology: Vero E6 (CL1008), the African green monkey kidney epithelial cell, were infected with SARS-CoV-2 virus (isolate USA-WA1/2020) in a 96 well-plate. NOQ19 test material was diluted at different concentration: 0.05 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml and 0.9 mg/ml. These different concentrations of NOQ19 were added to infected cells respectively and incubated for 3 days in 5% CO2 incubator. Remdesivir was used as a positive control. The cells were finally fixed with formaldehyde, stained with crystal violet and plaques were visualized. The number of plaques were counted to determine the PFU(plaque forming units)/ml. Results: The results of the present study demonstrated an excellent an antiviral efficacy of NOQ19 at 0.9 mg/ml concentration, eliminating 100% virus. The IC50 of the drug was found to be 0.2 mg/ml. Conclusion: There is limited data around pre-clinical efficacy of polyherbal Ayurvedic drugs. Ayurvedic and herbal formations need to be tested in a preclinical setting to support the human data. The results of the present study demonstrated viral load reduction using NOQ19 in Vero E6 cell lines infected with SARS-CoV-2 virus. These result along with other preclinical and clinical trials could further evaluate the efficacy of NOQ19 as a potential therapeutic option in the fighting the COVID-19 challenge.

A Sonochemical Access to 5-Aryl Substituted Pyrazolo[1,5-a]Pyrimidines as Potential Inhibitors of TNF-α

In spite of various and extensive studies known for pyrazolo[1,5-a]pyrimidines the synthesis, in silico studies and biological evaluation of their 5-(het)aryl analogs remained underexplored. The TNF-α inhibitors on the other hand has considerable therapeutic potential for autoimmune and inflammatory diseases in addition to cancer, diabetes and possibly COVID-19. In the current study 5-aryl pyrazolo[1,5-a]pyrimidines were explored as potential inhibitors of TNF-α that was supported by the in silico studies. This class of compounds was accessed via a sonochemical synthesis involving the acid catalyzed cyclocondensation reaction of aminopyrazoles with acrylophenones in the presence of aerial oxygen. The study indicated that the overall rate of the reaction was enhanced by ultrasound in the absence of which a longer duration and higher temperature was necessary. The current catalyst/promoter/ligand free and scalable method afforded a range of compounds. Some of these compounds showed good inhibition of TNF-α in vitro where ester/amide moiety at the C-3 position played a key role in interacting with the protein dimer as suggested by the in silico studies. Indeed, these groups formed H-bonds with A: GLY121 and B: TYR151 residues of TNF-α dimer in silico. A brief SAR within the series and in silico ADME/toxicity prediction for best active compounds is presented. Compounds 3a-c were identified as initial hits for further pharmacological evaluations. A sonochemical method has been developed for the facile synthesis of pyrazolo[1,5-a]pyrimidines that were evaluated as potential inhibitors of TNF-α. A sonochemical method has been developed for the facile synthesis of pyrazolo[1,5-a]pyrimidines that were evaluated as potential inhibitors of TNF-α.

In vivo efficacy and safety of Siddha medicine Kabasura Kudineer among COVID 19 infected Syrian golden hamsters. (preprint)

Background: The COVID-19 pandemic has overburdened current healthcare system and highlighted the need to explore potent remedies in Traditional medicine systems. Kabasura Kudineer (KSK), a poly herbal Siddha medicine, has shown great potential in treating COVID-19. Objective: The objective of the study is to explore the safety and efficacy of Kabasura Kudineer in a preclinical model for COVID-19: Syrian Golden Hamsters. Methods: This research study investigates the in vivo efficacy and safety of the well-known antiviral Siddha medicine KSK as a powdered tablet on COVID-19 infected Syrian golden hamsters. A total of 19 female hamsters were infected with the virus cell culture through intranasal route. 4 out of 19 animals were mock controls, 5 were infection controls, 4 were treated with remdesivir and acted as positive controls and remaining 6 were treated with KSK. The hamsters were observed for any adverse events, followed by their sacrifice on day 4 after inoculation with the virus. The lung pathology and viral load was studied for each hamster. Results: Therapeutic use of intraperitoneal instillation of Siddha formulation KSK reduces SARS-CoV-2 viral load and associated gross clinical parameters. Results showed significant reduction of 65% in the viral load for the KSK arm as compared to the infection control. Conclusion: We observed that the animals treated with KSK exhibited less severe pathology compared to the untreated infected group. No toxicity or adverse events were observed in the KSK group. This pre-clinical study supports the safety and efficacy of KSK.Study Registration: FNDR’s Institutional Animal Ethics Committee (IAEC), Registration Number 2082/PO/Rc/S/19/CPCSEA

AYUSH medicine as add-on therapy for mild category COVID-19; an open label randomised, controlled clinical trial. (preprint)

BackgroundAYUSH formulations have a potential role in symptomatic treatment, preventing disease progression and improving quality of life in COVID-19 patients. ObjectiveTo study the effect of AYUSH formulation (Kabasura Kudineer tablets, Shakti drops and Turmeric plus) as an add-on treatment in patients with mild COVID - 19 MethodologySingle centre, two arms, open labelled randomized controlled trial with a total of 30 patients (15 in the intervention arm and 15 in the standard care arm). Intervention arm received a combination of 3 AYUSH formulation along with the standard of care treatment for 21 days. All patients were followed for 28 days. Symptom severity (using Modified Jackson scale), negative conversion of SARS-CoV-2 RNA (using RTPCR) and quality of life (WHOWOL BREF questionnaire) was assessed. ResultsFifteen patients (93.8%) in the intervention group and twelve patients (92.3%) in the standard care arm had complete resolution of symptoms (P value= 0.36). Negative conversion for SARS-CoV-2 was seen in thirteen patients (92.9%) in intervention arm and eleven patients (100%) in standard care arm at day 28 (P value = 0.56). There was no difference in the quality of life scores between the 2 groups. ConclusionThe use of Ayush interventions as add-on therapy did not negatively impact the clinical outcomes in COVID-19. This trial confirmed the safety and tolerability of Kabasura Kudineer tablets, Shakti drops and Turmeric plus tablets when used use among mild to moderate symptom category, of COVID-19. There were no serious adverse events in the treated group. There was no clinical progression of disease from baseline status and all trial participants recovered fully by day 28. A longer follow up and a larger sample size is recommended for future definitive trials with this alternative medicine (AYUSH) combination.