Assessment of the Impact of the Egyvir on Rats Experimental Animals, a Preclinical Study for Sars-Cov-2 Treatment

Background: The SARS-COV-2 is a worldwide pandemic problem. We developed a herbal extract with potent in-vitro virucidal, anti-inflammatory and immunomodulatory effects called EGIVIR. Our aim is to assess the bioavailability and cytotoxicity of EGYVIR on different organs and biological systems in Sprague Dawley rats as a model of experimental animals.Methods: 128 rats were divided into 16 groups (8 rats each), where Egyvir was assessed in oral doses of 20, 30, and 40 mg/kg body weight, and by inhalation in 0.2, 0.3, and 0.4 mg/kg body weight, four times/day, compared to the control groups.Results: The Egyvir had no significant effect on the blood pressure, pulse, motor activity, histological, hematological, and coagulation profiles. Also, the blood levels of triglycerides, cholesterol, blood glucose, lactate dehydrogenase (LDH), and creatine phosphor kinase (CPK) were not significantly affected. Egyvir had no harmful effect on the kidney and liver functions, blood electrolytes levels and urinary levels of sodium, potassium, and chloride. There was no significant effect on the serum levels of interleukin-113 (IL -113), IL-2, IL-4, IL-6, IL-10, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha). Additionally, there was no significant change in the levels of Superoxide dismutase (SOD), catalase, reduced glutathione (GSH), and malonaldehyde (MDA) in comparison to the control groups (P<0.05).Conclusion: Egyvir is considered a safe antiviral natural drug. It could be used for the treatment of SARS-COV-2 without any adverse effects when used with the recommended doses. However, these data are a preliminary step for validation in a clinical setting.

Phytochemical constitution and antioxidant activity of functional herbal drink from Indian gooseberry (Emblica officinalis Gaertn.) fruits containing spices and condiments

Functional beverages originate primarily from fruits and vegetables sources, but also include those from other plants such as tea, coffee, cocoa, soybean as well as animal products like milk and dairy-based and alcoholic drinks. They have definite medical or health benefits which include prevention or delaying the progress of diseases. Indian gooseberry is a very rich source of vitamin C and phenolics, two potent antioxidant compounds. Similarly curcumin in turmeric, piperine in black pepper and gingerol in ginger have proven antioxidant and anti-inflammatory properties. Incidence of covid-19 pandemic has raised awareness among people the importance of maintaining higher levels of immunity. A study was undertaken at Kerala Agricultural University during 2020–21 to develop an herbal functional drink from Indian gooseberry fruit juice incorporated with turmeric and black pepper powders, ginger juice extract and juice of acid lime fruits. The herbal drink formulated with these ingredients was homogenized at an operating pressure of 175 Bar with a speed of 235 SPM and was subsequently pasteurized at 100 0 C for 10 minutes in glass bottles, followed by storage under refrigerated conditions at 5 ± 2 0 C for 3 months. The initial ascorbic acid, total phenolics, total flavonoids, total carotenoids and total curcumin contents were 61.0 mg100g− 1, 184.0 mg100g− 1, 153.0 mg100g− 1, 119.98 mg100g− 1 and 31.0 mg100g− 1, respectively. Antioxidant activity of the herbal drink was determined by three assays, viz. ABTS, DDPH and FRAP. The initial IC 50 values of the herbal drink by ABTS, DPPH and FRAP assays were 8.64, 0.212 and 0.368 μgml− 1, respectively. Significant decline in ascorbic acid, total flavonoids, total carotenoids and curcumin content were recorded in the product during storage in contrast to the total phenolics content which showed a significant rise over the storage period. Antioxidant activity of the herbal drink determined by all the three assays also declined significantly throughout the storage period. The results indicate that the product can be promoted as a healthy drink which has to be stored at low temperature in order to retain higher levels of antioxidant compounds and antioxidant activity. Graphical : [Figure not available: see fulltext.] © 2023, The Author(s).

Simultaneous Quantification of Piperine, Vasicine and Eugenol in Kabasura Kudineer by HPTLC

Kabasura Kudineer is a polyherbal decoction of the Siddha medical system (an Indian system of medicine), traditionally used to cure fever, colds, coughs, and respiratory ailments. The government of India had recommended Kabasura Kudineer as one of many preventive/treatment measures for COVID-19. Kabasura Kudineer Choornam is an admixed coarse powder of 15 herbs and its decoction is Kabasura Kudineer. The chemical constituents in the 15 herbs used for the preparation of the Choornam are known but the constituents present in the Kabasura Kudineer (decoction) are unidentified. Piperine, vasicine and eugenol are known for their potent activity against respiratory tract infections;hence, they were selected as marker compounds. The present work was planned to simultaneously quantify piperine, vasicine and eugenol in Kabasura Kudineer by the HPTLC method. The optimised mobile phase was toluene: ethyl acetate: methanol: ammonia (5:9:3:0.5, v/v/v/v), and the scanning was carried out at 287 nm. The Rf values of piperine, vasicine and eugenol were found to be 0.70, 0.32 and 0.82, respectively. The linearity range of piperine and vasicine was 500-3000 ng spot-1 and it was 10-60 ng spot-1 for eugenol. The quantities of piperine, vasicine and eugenol in Kabasura Kudineer (100 mL) were 0.03, 0.056 and 0.035 % w/v, respectively. This developed method can be used to simultaneously quantify piperine, vasicine and eugenol in any polyherbal formulation.Copyright © 2023, Informatics Publishing Limited and Society for Biocontrol Advancement. All rights reserved.

Molecular Insights into Resveratrol and Its Analogs as SARS-CoV-2 (COVID-19) Protease Inhibitors

Introduction: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic is plaguing the entire world. Amidst the pandemic, research and development efforts are fo-cused on the challenges associated with the SARS-CoV-2 structure. Material(s) and Method(s): Efficient computational methodologies are applied to screen the available FDA-approved drugs/datasets/libraries to identify potent molecules. In the present study, we have carried out ab initio quantum chemical studies, including relativistic effects followed by molecular docking with the SARS-CoV-2 protease target by employing a tailor-made library consisting of molecular analogs of Resveratrol, a natural bioflavonoid. Result(s): The derived docking results were validated with ab initio quantum computations that in-cluded both density functional level (DFT) and Moller-Plesset second order perturbation theories (MP2). We found to be that Resveratrol and its analogs (R8 and R17) bind to the SARS-CoV-2 protease target. In addition to this, the computed IR spectrum is found in agreement with the report-ed experimental spectra for Resveratrol complexes and thus validates the modeling and reliability of proposed geometries. The solvation energies in the aqueous phase obtained using enhanced aug-cc-pVTZ basis sets confirm enhancement of bioavailability for Resveratrol through piperine, a natural alkaloid. Conclusion(s): The potential of the natural bioflavonoid Resveratrol and its analogs to be investigated through in vivo and in vitro SARS-CoV-2 protease models is concluded. The study investigated the potential of natural polyphenols as promising anti-viral therapeutics.Copyright © 2021 Bentham Science Publishers.

Molecular dynamics and absolute binding free energy studies of piperine derivatives as potential inhibitors of SARS-CoV-2 main protease.

The work presents a library of piperine derivatives as potential inhibitors of the main protease protein (Mpro) functionality using Docking Studies, Molecular Dynamics (MD) Simulations and Absolute Binding Free-Energy calculations. 342 ligands were selected for this work and docked with Mpro protein. Among all the ligands studied, PIPC270, PIPC299, PIPC252, PIPC63, PIPC311 were the top five docked conformations having significant hydrogen bonding and hydrophobic interactions inside the active pocket of Mpro. These top five ligands were subjected to MD simulations for 100 ns using GROMACS. Root Mean Square Deviation (RMSD), Root Mean Square Fluctuation (RMSF), Radius of Gyration (Rg), Solvent Accessible Surface Area (SASA) and hydrogen bond analysis revealed that the ligands bounded to protein remain stable without significant deviations during the course of MD simulations. Absolute binding free energy (ΔGb) was calculated for theses complexes and found that the ligand PIPC299 shows the prevalent binding affinity with binding free-energy of about -113.05 Kcal/mol. Thus, these molecules can be further tested by in vitro and in vivo studies on Mpro. This study lays a path to explore the new functionality of piperine derivatives as novel drug like molecules.Communicated by Ramaswamy H. Sarma.

Curcumin-piperine co-supplementation and human health: A comprehensive review of preclinical and clinical studies.

Curcumin is extracted from the rhizomes Curcuma longa L. It is known for its anti-inflammatory and anti-oxidant activities. Despite its safety and potential for use against various diseases, curcumin's utility is restricted due to its low oral bioavailability. Co-administration of curcumin along with piperine could potentially improve the bioavailability of curcumin. The present review aimed to provide an overview of the efficacy and safety of curcumin-piperine co-supplementation in human health. The findings of this comprehensive review show the beneficial effects of curcumin-piperine in improving glycemic indices, lipid profile and antioxidant status in diabetes, improving the inflammatory status caused by obesity and metabolic syndrome, reducing oxidative stress and depression in chronic stress and neurological disorders, also improving chronic respiratory diseases, asthma and COVID-19. Further high-quality clinical trial studies are needed to firmly establish the clinical efficacy of the curcumin-piperine supplement.

LIVER INJURY ASSOCIATED WITH TURMERIC&≤x2013;A GROWING PROBLEM: NINE CASES FROM THE DRUG-INDUCED LIVER INJURY NETWORK [DILIN]

BACKGROUND: Turmeric (curcumin) is a commonly used over-the-counter herbal product whose uses include diarrhea, arthritis, cancer and even COVID-19. Recently turmeric has been implicated in cases of clinically apparent liver injury with jaundice. The aim of this case series is to describe the clinical, histologic and human leukocyte antigen (HLA) associations of turmeric-associated hepatotoxicity as seen in the U.S. Drug Induced Liver Injury Network (DILIN) Prospective Study. METHODS: All adjudicated cases enrolled in DILIN between 2003-2020 with turmeric as an implicated product were reviewed. Causality was assessed using a 5-point expert opinion score. Available products were collected and analyzed for the presence of turmeric using ultra-high-performance liquid chromatography. Genetic analyses included HLA sequencing. RESULTS: Of 1697 cases of drug-induced liver injury judged to be definite, highly likely or probable (high confidence), nine (0.5%) were attributed to turmeric, all of which were enrolled since 2012, and 6 since 2017 (Figure). The 9 cases included 7 women, 8 whites, with a mean age of 51 years (range, 35-62 years) and BMI 25 kg/m2 (range, 15-40). Seven patients used alcohol, but none to excess, and none had underlying liver disease. Turmeric was used for an average of 102 days before onset of injury (range, 30-425 days). Initial mean ALT was 1179 U/L (range, 328-2245), ALP 211 U/L (41-441), total bilirubin 5.9 mg/dL (1.2-10.8), and INR 1.0 (0.9-1.2). Six patients developed jaundice, and serum bilirubin peaked at 9.6 mg/dL (0.8-26), and INR 2.3 (1.0- 9.7). Liver injury was hepatocellular in 8 patients (mean R = 22). Five patients had elevated antinuclear antibody (ANA) titer and two anti-smooth muscle (ASM) antibody, but none were treated with corticosteroids. Liver biopsy in 5 patients showed portal and lobular mixed inflammatory infiltrates with lymphocytes and eosinophils typical of drug-induced liver injury. Five patients were hospitalized, and one patient died of acute liver failure. Chemical analysis confirmed the presence of turmeric in all 7 products analyzed;3 also contained piperine (black pepper), and none contained green tea. Of 7 patients with HLA typing available, 4 carried HLA-B*35:01, a class I HLA allele previously implicated in both green tea and Polygonum multiflorum hepatotoxicity. CONCLUSION: Liver injury due to turmeric appears to be increasing, perhaps, reflecting usage patterns or increased combination with black pepper, which increases its absorption. Turmeric liver injury, similar to that caused by other polyphenolic herbal products, is typically hepatocellular, with a latency of 1 to 6 months, and is linked to HLA-B*35:01. While most cases are self-limited, the injury can be severe and result in death or liver transplantation.

A Computational-Experimental Investigation of the Molecular Mechanism of Interleukin-6-Piperine Interaction.

Herein, we elucidate the biophysical aspects of the interaction of an important protein, Interleukin-6 (IL6), which is involved in cytokine storm syndrome, with a natural product with anti-inflammatory activity, piperine. Despite the role of piperine in the inhibition of the transcriptional protein NF-κB pathway responsible for activation of IL6 gene expression, there are no studies to the best of our knowledge regarding the characterisation of the molecular interaction of the IL6-piperine complex. In this context, the characterisation was performed with spectroscopic experiments aided by molecular modelling. Fluorescence spectroscopy alongside van't Hoff analyses showed that the complexation event is a spontaneous process driven by non-specific interactions. Circular dichroism aided by molecular dynamics revealed that piperine caused local α-helix reduction. Molecular docking and molecular dynamics disclosed the microenvironment of interaction as non-polar amino acid residues. Although piperine has three available hydrogen bond acceptors, only one hydrogen-bond was formed during our simulation experiments, reinforcing the major role of non-specific interactions that we observed experimentally. Root mean square deviation (RMSD) and hydrodynamic radii revealed that the IL6-piperine complex was stable during 800 ns of simulation. Taken together, these results can support ongoing IL6 drug discovery efforts.

The efficacy of curcumin-piperine co-supplementation on clinical symptoms, duration, severity, and inflammatory factors in COVID-19 outpatients: a randomized double-blind, placebo-controlled trial.

BACKGROUND: COVID-19 pandemic has made the disease a major global problem by creating a significant burden on health, economic, and social status. To date, there are no effective and approved medications for this disease. Curcumin as an anti-inflammatory agent can have a positive effect on the control of COVID-19 complications. This study aimed to assess the efficacy of curcumin-piperine supplementation on clinical symptoms, duration, severity, and inflammatory factors in patients with COVID-19. METHODS: Forty-six outpatients with COVID-19 disease were randomly allocated to receive two capsules of curcumin-piperine; each capsule contained 500 mg curcumin plus 5 mg piperine or placebo for 14 days. RESULTS: Mean changes in complete blood count, liver enzymes, blood glucose levels, lipid parameters, kidney function, and c-reactive protein (CRP) were not significantly different between the two groups. There was a significant improvement in health status, including dry cough, sputum cough, ague, sore throat, weakness, muscular pain, headache, and dyspnea at week 2 in both curcumin-piperine and placebo groups (P value < 0.05); however, the improvement in weakness was more in the curcumin-piperine group than with placebo group (P value 025). CONCLUSION: The present study results showed that curcumin-piperine co-supplementation in outpatients with COVID-19 could significantly reduce weakness. However, in this study, curcumin-piperine co-supplementation could not significantly affect the other indices, including biochemical and clinical indices. TRIAL REGISTRATION: Iranian Registry of Clinical Trials IRCT20121216011763N46 . 2020-10-31.

Molecular and pharmacological aspects of piperine as a potential molecule for disease prevention and management: evidence from clinical trials.

BACKGROUND: Piperine is a type of amide alkaloid that exhibits pleiotropic properties like antioxidant, anticancer, anti-inflammatory, antihypertensive, hepatoprotective, neuroprotective and enhancing bioavailability and fertility-related activities. Piperine has the ability to alter gastrointestinal disorders, drug-metabolizing enzymes, and bioavailability of several drugs. The present review explores the available clinical and preclinical data, nanoformulations, extraction process, structure-activity relationships, molecular docking, bioavailability enhancement of phytochemicals and drugs, and brain penetration properties of piperine in the prevention, management, and treatment of various diseases and disorders. MAIN BODY: Piperine provides therapeutic benefits in patients suffering from diabetes, obesity, arthritis, oral cancer, breast cancer, multiple myeloma, metabolic syndrome, hypertension, Parkinson's disease, Alzheimer's disease, cerebral stroke, cardiovascular diseases, kidney diseases, inflammatory diseases, and rhinopharyngitis. The molecular basis for the pleiotropic activities of piperine is based on its ability to regulate multiple signaling molecules such as cell cycle proteins, anti-apoptotic proteins, P-glycoprotein, cytochrome P450 3A4, multidrug resistance protein 1, breast cancer resistance protein, transient receptor potential vanilloid 1 proinflammatory cytokine, nuclear factor-κB, c-Fos, cAMP response element-binding protein, activation transcription factor-2, peroxisome proliferator-activated receptor-gamma, Human G-quadruplex DNA, Cyclooxygenase-2, Nitric oxide synthases-2, MicroRNA, and coronaviruses. Piperine also regulates multiple signaling pathways such as Akt/mTOR/MMP-9, 5'-AMP-activated protein kinase-activated NLR family pyrin domain containing-3 inflammasome, voltage-gated K+ current, PKCα/ERK1/2, NF-κB/AP-1/MMP-9, Wnt/ß-catenin, JNK/P38 MAPK, and gut microbiota. SHORT CONCLUSION: Based on the current evidence, piperine can be the potential molecule for treatment of disease, and its significance of this molecule in the clinic is discussed.

Semi-Synthesis of N-Aryl Amide Analogs of Piperine from Piper nigrum and Evaluation of Their Antitrypanosomal, Antimalarial, and Anti-SARS-CoV-2 Main Protease Activities.

Piper nigrum, or black pepper, produces piperine, an alkaloid that has diverse pharmacological activities. In this study, N-aryl amide piperine analogs were prepared by semi-synthesis involving the saponification of piperine (1) to yield piperic acid (2) followed by esterification to obtain compounds 3, 4, and 5. The compounds were examined for their antitrypanosomal, antimalarial, and anti-SARS-CoV-2 main protease activities. The new 2,5-dimethoxy-substituted phenyl piperamide 5 exhibited the most robust biological activities with no cytotoxicity against mammalian cell lines, Vero and Vero E6, as compared to the other compounds in this series. Its half-maximal inhibitory concentration (IC50) for antitrypanosomal activity against Trypanosoma brucei rhodesiense was 15.46 ± 3.09 µM, and its antimalarial activity against the 3D7 strain of Plasmodium falciparum was 24.55 ± 1.91 µM, which were fourfold and fivefold more potent, respectively, than the activities of piperine. Interestingly, compound 5 inhibited the activity of 3C-like main protease (3CLPro) toward anti-SARS-CoV-2 activity at the IC50 of 106.9 ± 1.2 µM, which was threefold more potent than the activity of rutin. Docking and molecular dynamic simulation indicated that the potential binding of 5 in the 3CLpro active site had the improved binding interaction and stability. Therefore, new aryl amide analogs of piperine 5 should be investigated further as a promising anti-infective agent against human African trypanosomiasis, malaria, and COVID-19.

In silico and Molecular Docking Studies of Black Pepper Phyto-constituents against EmrD Efflux Pump of E. coli

Multidrug resistance (MDR) bacterial infection is the next pandemic waiting behind the COVID-19 with annual mortality rate 700000 worldwide. Among the MDR bacteria, Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterococcus faecium and Enterococcus faecalis are showing average resistance of 50 to 80% to ampicillin, amoxicillin, third-generation cephalosporin’s and fluoroquinolone and even to combinations antibiotics such as amoxicillin-clavulanic acid. To make the antibiotic resistance issue worst, pharmaceutical industry is reluctant to invest in research and development of new antibiotic typically due to low returns on investment. Accordingly, use of combination of two or more antibiotics or use of the antibiotic adjuvants are only available ways in combatting the ever growing Multidrug resistance (MDR) in bacteria. The present paper is designed to analyze the synergistic potential of black pepper phyto-constituents as the amoxicillin adjuvants in comparison with isolated piperine against the MDR E. coli. using in-silico molecular docking. The result indicates that binding energy (Kcal/mol) and torsion free energy (Kcal/mol) of piperine (-6.23, +0.89), beta caryophyllene (-6.36, +0.00), beta selinene (-6.93, +0.30), beta-Thujene (-5.42, +0.30) is less for the emrD efflux pump as compared to amoxicillin (-5.85, +2.93) respectively indicating strong inhibition for EmrD of MDR E-coli than amoxicillin. The results are also indicating that black pepper extract containing all aforementioned phyto-constituents has synergistic effect in comparison with isolated piperine against the MDR E. coli. ADMET of these phyto-constituents also indicates their safety profile in combination with amoxicillin.

Curcumin and Piperine in COVID-19: A Promising Duo to the Rescue?

COVID-19 is now pandemic throughout the world, and scientists are searching for effective therapies to prevent or treat the disease. The combination of curcumin and piperine is a potential option for the management of COVID-19 based on several mechanisms including antiviral, anti-inflammatory, immunomodulatory, antifibrotic, and antioxidant effects. Here, we describe the probable mechanism of curcumin-piperine against COVID-19. Administration of curcumin-piperine combination appears as a potential strategy to counterbalance the pathophysiological features of COVID-19 including inflammation. The optimal dose and duration of curcumin-piperine supplementation should be determined in the future.

Statistical optimization of bile salt deployed nanovesicles as a potential platform for oral delivery of piperine: accentuated antiviral and anti-inflammatory activity in MERS-CoV challenged mice.

The objective of this paper is to confine piperine, a poor oral bioavailable herbal drug into bile salt based nano vesicles for improving its aqueous solubility, hence, its therapeutic activity. Piperine-loaded bilosomes were fabricated adopting thin film hydration technique according to 32.21 full factorial design to investigate the impact of different formulation variables on the characters of bilosomes: entrapment efficiency (EE%), particle size, and % of drug released post 8 h (Q8hr). The selected optimum formula was F2 (enclosing 1% bile salt, brij72 as a surfactant, and ratio of surfactant:cholesterol was 9:1) with desirability value 0.801, exhibiting high EE% (97.2 ± 0.8%) nanosized spherical vesicles (220.2 ± 20.5 nm) and Q8hr (88.2%±5.6). The superiority of the optimized formula (F2) over the drug suspension was revealed via ex vivo permeation study, also pharmacokinetic study denoted to the boosted oral bioavailability of piperine-loaded bilosome compared to piperine suspension. Moreover, antiviral activity and safety margin of F2 was significantly higher than that of the drug suspension. The ability of piperine to interact with the key amino acids in the receptor binding domain 4L3N as indicated by its docking configuration, rationalized its observed activity. Furthermore, F2 significantly reduce oxidant markers, inflammatory cytokines in MERS-CoV-infected mice. Hence, bilosomes can be considered as a carrier of choice for piperine with potential antiviral and anti-inflammatory activities.

Effects of curcumin-piperine co-supplementation on clinical signs, duration, severity, and inflammatory factors in patients with COVID-19: a structured summary of a study protocol for a randomised controlled trial.

OBJECTIVES: This study aims to investigate the efficacy of curcumin-piperine co-supplementation on disease duration, severity and clinical symptoms, and inflammatory mediators in patients with coronavirus (COVID-19). TRIAL DESIGN: This is a randomized, placebo-controlled, double-blind, parallel arm clinical trial. PARTICIPANTS: All patients aged 20-75 years with the diagnosis of Covid-19 based on the PCR test. The exclusion criteria will include an age less than 20 and more than 75 years, current use of warfarin or other anticoagulant drugs, and the presence of sensitivity to herbal products such as turmeric and pepper. This study will be conducted in academic hospitals affiliated to Isfahan University of Medical Sciences, Isfahan, Iran. INTERVENTION AND COMPARATOR: Fifty outpatients will be randomly allocated in a ratio of 1:1 to receive a capsule of curcumin-piperine containing 500 mg curcumin plus 5 mg piperine or matching placebo containing 505 mg maltodextrin twice a daily, after lunch and dinner, over a period of 2 weeks. Similarly, 50 inpatients who are admitted to hospital wards excluding intensive care unit (ICU) will be randomly assigned in a ratio of 1:1 to receive a capsule curcumin-piperine or matching placebo (provided by the Sami Labs company) twice a daily, after lunch and dinner, over a period of 2 weeks. MAIN OUTCOMES: The main outcomes of this study are the efficacy of curcumin-piperine on coronavirus disease's clinical symptoms, duration, severity, and inflammatory mediators after 2 weeks of curcumin-piperine co-supplementation. RANDOMISATION: Randomization sequences will be generated with the use of a random-number table with a permuted block design (block size of 4) and stratification according to the gender variable (male vs. female). These sequences will be prepared by an independent statistician and will be kept in opaque, sealed, numbered envelopes which will be opened only at the time of enrollment. The allocation ratio in intervention and control groups is 1:1. Researchers and all patients will be unaware of the study-group assignment until the completion of data analyses. BLINDING (MASKING): This study is a double-blind clinical trial (participant, researcher). The curcumin-piperine and placebo supplements are packaged in similar numbered drug containers, and the researcher and all patients will be unaware of the study assignment until the end of the study. NUMBERS TO BE RANDOMISED (SAMPLE SIZE): The calculated total sample size is 100 patients, with 25 patients in each group. TRIAL STATUS: The protocol is Version 2.0, May 24, 2020. Recruitment began May 4, 2020, and is anticipated to be completed by April 19, 2021. TRIAL REGISTRATION: This trial has been registered by the title of "Effect of curcumin-piperine co-supplementation on disease duration, severity and clinical signs, and inflammatory factors in patients with coronavirus (COVID-19): A randomized, double-blind, placebo-controlled clinical trial study" in the Iranian Registry of Clinical Trials (IRCT) with code "IRCT20121216011763N46", https://www.irct.ir/trial/47529 . The registration date is May 4, 2020. FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.

Spice-infused palmyra palm syrup improved cell-mediated immunity in Wistar Albino rats.

Spices attract tremendous attention in the management of viral infections. However, scientific validation is vital to recommend spices as nutraceuticals or functional foods. In the present work, we have selected three spices based on Ayurvedic knowledge and developed a nutraceutical for immunomodulation. Trikatu, a blend of ginger, black pepper, and long pepper, is used in the Indian Ayurvedic system, along with many herbs, for various ailments. We formulated a "Trikatu syrup" (TS) using these three spices and palmyra palm neera. Carbon clearance assay, neutrophil adhesion test, and sheep red blood cell (SRBC)-induced delayed-type hypersensitivity (DTH) reaction was performed to investigate the immunomodulatory potential of TS in Wistar Albino rats. The rats fed with TS showed a dose-dependent increase in footpad thickness compared to control rats, suggesting cell-mediated immunity. The major bioactive piperine in TS was isolated and quantified. PRACTICAL APPLICATIONS: Spices are consumed worldwide as a flavor enhancer in food. Besides, spices have an array of bioactive molecules with a multitude of health benefits. In the backdrop of COVID-19, immunomodulation and antiviral properties of spices are discussed widely. The present study is intended to explore the potential of three selected spices (ginger, black pepper, and long pepper) beyond its application in typical food preparations. The syrup formulated in this study by using these three spices improved cell-mediated immunity in Wistar Albino rats. The study warrants further validation studies of the formulated product for providing indisputable claims for the immunomodulation properties.