Probenecid

Initial Development and Early Therapeutic Rationale for Probenecid

This compound, a sulfonamide derivative, was synthesized in 1949 by Miller, Ziegler, and Sprague with the primary objective of decreasing the renal clearance of antibiotics, particularly penicillin. nih.govnih.gov It was introduced to the market in 1950. nih.gov The rationale behind this development was to increase and prolong the plasma concentrations of co-administered antibiotics, thereby enhancing their effectiveness and allowing for lower dosing, which was especially relevant during periods of limited antibiotic supply, such as World War II. nih.govnewdrugapprovals.org

Shifting Clinical Relevance and Research Focus of this compound

Despite its initial importance in antibiotic therapy and its approval for gout treatment, the clinical relevance of this compound has significantly declined over the years. nih.govresearchgate.netnih.gov The expanded production of more diverse, cheaper, and safer antibiotics reduced the necessity of using this compound to boost antibiotic levels for many common infections in the post-World War II era. nih.govresearchgate.net In the treatment of gout, the introduction of alternative therapies, such as allopurinol in the 1960s, which inhibits uric acid production, and the use of colchicine and steroids for acute attacks, diminished this compound's role as a primary treatment. researchgate.netexplorationpub.com While still approved for gout and hyperuricemia, it is rarely used today for either indication compared to its past prevalence. researchgate.netexplorationpub.com Its efficacy in lowering serum uric acid is considered moderate, and it may not be effective in patients with significant renal impairment. explorationpub.commedsafe.govt.nz

However, the decline in its traditional clinical use has coincided with a shifting research focus, exploring other pharmacological properties and potential applications. nih.govresearchgate.netnih.gov

Emergence of New Therapeutic Potentials and Research Avenues for this compound

In recent years, research into this compound has revealed its ability to interact with various membrane proteins beyond the well-known OATs and URAT1, suggesting new potential therapeutic utilities. nih.govmdpi.comresearchgate.net These interactions include the blockade of pannexin-1 (Panx1) hemichannels and the activation of Transient Receptor Potential Vanilloid 2 (TRPV2) channels. nih.govresearchgate.netmdpi.com

The blockade of Panx1 hemichannels has garnered attention for its potential role in reducing neuroinflammation, a key factor in various central nervous system (CNS) disorders. nih.govmdpi.comresearchgate.net Panx1 hemichannels are involved in the release of ATP and other molecules that contribute to inflammatory responses and affect neuronal communication. mdpi.com By inhibiting these channels, this compound may exert neuroprotective, antiepileptic, and anti-inflammatory effects, which are being investigated in the context of conditions like ischemia, sepsis, epilepsy, Parkinson's, and Alzheimer's diseases. nih.govmdpi.comresearchgate.net Furthermore, this compound's ability to inhibit organic anion and cation transporters in the brain (OATs and OCTs) can increase the bioavailability of certain drugs in the CNS, suggesting its potential as an adjuvant therapy to improve the effectiveness of conventional treatments for psychiatric and neurological disorders where drug delivery to the brain is a challenge. nih.govmdpi.comresearchgate.net

The identification of this compound as an agonist of TRPV2 channels has opened up research avenues in cardiology and neurology. researchgate.netnih.govnih.gov TRPV2 channels are involved in various physiological processes, including calcium signaling. nih.govmdpi.com Studies have shown that this compound can exert a positive inotropic effect on cardiomyocytes, mediated by TRPV2 channels, suggesting a potential role in treating heart conditions. nih.gov

Beyond these specific targets, this compound's interaction with OATs and its effect on drug excretion continue to be relevant in research for optimizing the pharmacokinetics of other medications, including some antivirals like oseltamivir, aciclovir, ganciclovir, and zidovudine, as well as certain nonsteroidal anti-inflammatory drugs (NSAIDs) and methotrexate. medicinesinformation.co.nzmedsafe.govt.nzebi.ac.ukwikipedia.orgspandidos-publications.com This property is being explored to enhance the systemic exposure and efficacy of these drugs or to reduce their renal toxicity. newdrugapprovals.orgmedicinesinformation.co.nzmedsafe.govt.nz

Modulation of Organic Anion Transporters (OATs) by Probenecid

This compound is a well-established inhibitor of organic anion transporters (OATs), a family of proteins crucial for the transport of a wide range of endogenous and exogenous organic anions across cell membranes. nih.govpatsnap.com OATs are predominantly located in the kidneys and liver, playing a significant role in the body's elimination of toxins and drugs. patsnap.com this compound's interaction with OATs is primarily characterized by competitive inhibition, where it competes with other organic anions for binding sites on the transporter proteins. patsnap.compatsnap.com

Modulation of Transient Receptor Potential Vanilloid 2 (TRPV2) Channels by this compound

Beyond its effects on organic anion transporters, this compound has also been identified as a modulator of Transient Receptor Potential Vanilloid 2 (TRPV2) channels. taylorandfrancis.comnih.govcellphysiolbiochem.commendeley.comguidetopharmacology.orgnih.gov TRPV2 is a non-selective cation channel expressed in various tissues, including the heart, neurons, and immune cells. mdpi.comnih.govbiorxiv.org

Modulation of Pannexin-1 (Panx1) Hemichannels by this compound

Pannexin-1 (Panx1) channels are ATP-permeable channels expressed in various cell types, including neurons and glial cells mdpi.comroyalsocietypublishing.org. They play a role in paracrine cell communication through the release of ATP into the extracellular space mdpi.com. This extracellular ATP can activate purinergic receptors, such as P2X7 receptors (P2X7R), which can further influence cellular processes, including calcium influx and inflammasome activation mdpi.comresearchgate.netdovepress.comnih.gov.

This compound has been identified as an inhibitor of Panx1 channels nih.govnih.gov. Studies have shown that this compound can inhibit voltage-induced Panx1 currents in experimental settings researchgate.net. The inhibition of Panx1 by this compound is concentration-dependent nih.gov. While this compound is considered a Panx1 inhibitor, it is important to note that it may require millimolar concentrations for this effect and has been reported to block other ion channels as well biorxiv.org. Some studies suggest that this compound's effect on calcium release may be independent of its Panx1 blocking activity rupress.org.

Table 1: this compound Inhibition of Pannexin-1 Channels

Panx1 hemichannels are involved in the release of ATP from cells mdpi.comroyalsocietypublishing.org. This released ATP can contribute to inflammatory mechanisms, including the activation of the inflammasome researchgate.netdovepress.comelifesciences.org. The inflammasome is a multiprotein complex that plays a key role in innate immunity and leads to the processing and release of inflammatory cytokines like IL-1β and IL-18 researchgate.netdovepress.comnih.gov.

Research suggests that this compound, by blocking Panx1 channels, can affect ATP release and subsequently modulate inflammasome activation researchgate.netdovepress.comnih.govfrontiersin.org. Studies have shown that this compound can inhibit inflammasome activation in various cell types, including neurons and astrocytes nih.govfrontiersin.org. This inhibition of inflammation through Panx1 blockade may contribute to potential therapeutic effects of this compound in conditions involving neuroinflammation mdpi.comresearchgate.net.

Table 2: this compound's Impact on ATP Release and Inflammasome Activation

Inhibition of Multidrug Resistance-Associated Proteins (MRPs) by this compound

Multidrug Resistance-Associated Proteins (MRPs), members of the ATP-binding cassette (ABC) transporter family, are involved in the efflux of a wide range of substrates, including organic anions, conjugated drugs, and xenobiotics oup.comresearchgate.net. They play a significant role in drug resistance in various contexts, including cancer and the transport of endogenous metabolites oup.comresearchgate.net.

This compound has been identified as an inhibitor of MRPs oup.comtocris.com. Specifically, it has been shown to inhibit organic acid transport mediated by MRPs, such as MRP1 and OAT3 researchgate.nettocris.comnih.govnih.gov. This inhibition can affect the cellular accumulation and excretion of various compounds, including certain antibiotics and endogenous substances drugbank.comwikipedia.orgpatsnap.comnih.gov. While this compound can inhibit several MRPs, its inhibitory profile can vary among different MRP subtypes oup.com.

Potential Interaction with Other Molecular Targets of this compound

Beyond its established interactions with organic anion transporters and pannexin-1 channels, research using techniques like systems pharmacology and molecular docking has suggested potential interactions of this compound with other molecular targets nih.govfrontiersin.org. These findings offer insights into additional possible mechanisms underlying this compound's effects.

AKT1 (Protein Kinase B alpha) is a serine/threonine kinase involved in various cellular processes, including cell survival, proliferation, and metabolism spandidos-publications.com. It is a key component in signal transduction pathways.

Studies utilizing molecular docking and simulations have indicated that this compound may exhibit binding affinity with AKT1 nih.govfrontiersin.orgresearchgate.net. These computational analyses suggest a potential interaction, although further experimental validation is typically required to confirm the functional implications of such binding. AKT1 has been identified as one of the hub targets in network analysis exploring the pharmacological mechanisms of this compound in specific conditions nih.govfrontiersin.org.

Albumin (ALB) is the most abundant protein in plasma and plays crucial roles in transporting various molecules, maintaining oncotic pressure, and modulating the activity of other substances drugbank.comnih.gov.

Research, including systems pharmacology analyses, has identified Albumin as a potential interaction partner for this compound nih.govfrontiersin.orgresearchgate.net. This compound is known to have a high percentage of binding to plasma proteins, specifically binding significantly to albumin drugbank.comnih.gov. This binding can influence the pharmacokinetics of this compound and other drugs that also bind to albumin drugbank.comnih.gov.

Table 3: Potential Molecular Targets of this compound Identified Through Research

Absorption and Distribution Studies of Probenecid

Following oral administration, this compound is reported to be completely absorbed by the intestinal tract. mdpi.commedsafe.govt.nz Peak plasma levels are typically reached within two to four hours after oral administration. medsafe.govt.nz this compound exhibits a high degree of plasma protein binding, primarily to albumin, with reported binding percentages ranging from 85% to 95%. mdpi.commedsafe.govt.nz The apparent volume of distribution has been reported as approximately 11 liters or 0.003–0.014 L/kg. mdpi.commedsafe.govt.nznih.gov Tissue concentrations of this compound are generally lower than plasma concentrations, based on animal studies. nih.gov

Metabolism and Excretion Pathways of this compound

This compound undergoes biotransformation primarily in the liver. mdpi.com Metabolism involves both phase 1 oxidation of the alkyl side chains (approximately 70%) and phase 2 conjugation with glucuronic acid (approximately 20%). mdpi.com The major metabolites, including the acyl glucuronide, retain the ability to block the tubular secretion of organic acids but exhibit lower binding affinity to plasma proteins compared to the parent drug. mdpi.commedsafe.govt.nz The acyl glucuronide metabolite accounts for a significant portion of the dose, close to 50%. medsafe.govt.nz Other metabolites, such as mono-n-propyl, secondary alcohol, and carboxylic acid derivatives, are also formed, totaling about 30% of the dose. medsafe.govt.nznih.gov

Elimination of this compound and its metabolites occurs mainly via the kidneys. mdpi.comnih.gov Renal excretion is the primary route for metabolites, while excretion of the unchanged parent drug is minimal and dependent on urinary pH. nih.gov Approximately 75-88% of the administered dose is recovered in the urine, predominantly as metabolites. mdpi.commedsafe.govt.nz Studies have indicated that the elimination process involves parallel pathways, with the excretion of the major metabolite following Michaelis-Menten kinetics and oxidized metabolites adhering to pseudo-first-order kinetics influenced by product inhibition. nih.gov Renal clearance accounts for a notable portion of the total plasma clearance of this compound. nih.gov

Pharmacokinetic and Pharmacodynamic Interactions of this compound with Other Agents

This compound is well-known for its ability to interact with the disposition of other drugs, primarily by inhibiting renal tubular transport of organic acids. drugbank.comnih.govmedicinesinformation.co.nz This competitive inhibition, particularly of organic anion transporters (OATs) in the kidney, leads to increased plasma concentrations and prolonged half-lives of coadministered drugs that are substrates for these transporters. drugbank.commedicinesinformation.co.nz this compound also has been shown to inhibit glucuronidation, which can further influence the concentrations of drugs metabolized by this pathway. medicinesinformation.co.nznih.gov

Neuroprotective Properties of Probenecid in Animal Models

Preclinical studies utilizing animal models of neurological disorders have provided evidence for the neuroprotective effects of this compound. For instance, in the N171-82Q transgenic mouse model of Huntington's disease (HD), this compound administration was observed to improve motor behavior, increase cell survival, and significantly reduce neuronal loss and the number of intranuclear neuronal aggregates. mdpi.comnih.gov this compound has also shown protective effects in a rat model of cognitive impairment and in models of spinal cord injury (SCI). frontiersin.org In SCI models in SD rats, this compound reduced the lesion area and demyelination, and increased the survival of motor neurons, leading to improved locomotor function. frontiersin.org These findings suggest that this compound may hold therapeutic potential for various central nervous system (CNS) injuries and neurodegenerative conditions. mdpi.comnih.govfrontiersin.org

Anti-inflammatory Properties of this compound

This compound exhibits significant anti-inflammatory properties, which are relevant to its potential therapeutic uses in various conditions, including those affecting the CNS. mdpi.comnih.govfrontiersin.orgmdpi.comnih.gov

Antiviral Activity of this compound

Recent preclinical and translational research has highlighted the potent antiviral activity of this compound against a range of viruses, particularly respiratory viruses. researchgate.netmdpi.comnih.govnih.govmdpi.comfrontiersin.org Studies have demonstrated that this compound can inhibit the replication of SARS-CoV-2 (including various variants), influenza virus (including HPAI H5N1 and H7N9 strains), and respiratory syncytial virus (RSV) both in vitro and in animal models. researchgate.netmdpi.comnih.govnih.govmdpi.com

The antiviral mechanism of this compound appears to be linked to its interaction with host cell pathways rather than directly targeting viral components, which may reduce the likelihood of drug resistance. researchgate.netnih.govmdpi.com One proposed mechanism involves the solute carrier (SLC) superfamily, specifically its function as organic anion transporters (OATs). nih.govmdpi.com this compound's inhibition of OATs may disrupt viral replication processes that rely on these transporters. nih.gov Additionally, this compound's modulation of MAPK signaling pathways, such as the inhibition of JNK signaling, has been implicated in its antiviral effects against viruses like RSV. researchgate.net Furthermore, studies suggest that this compound may exert antiviral effects through coordinated modulation of pathways like ACE2 and PANX1, for example, by downregulating ACE2 expression, which is utilized by SARS-CoV-2 for entry. frontiersin.org In animal models, this compound treatment has been shown to reduce viral titers in the lungs and improve outcomes in infected mice. mdpi.comnih.gov

Cardiovascular Research with this compound

Beyond its antiviral properties, this compound has been investigated for its effects on the cardiovascular system, particularly in the context of heart failure. Research suggests that this compound can influence myocardial function through its interaction with calcium channels and its impact on contractility and relaxation.

Osteoclastogenesis Inhibition by this compound

Research has explored the potential of this compound to inhibit osteoclast formation, a process crucial for bone resorption. Excessive osteoclast differentiation is implicated in pathological bone diseases such as rheumatoid arthritis and osteoporosis. nih.gov Studies have investigated whether this compound can influence osteoclast formation, particularly through its effects on reactive oxygen species (ROS) formation in cellular models like RAW264.7 cells. nih.govbiomolther.orgsemanticscholar.org

Investigations have shown that this compound can dose-dependently reduce lipopolysaccharide (LPS)-induced ROS levels in RAW264.7 cells. nih.govbiomolther.orgsemanticscholar.org Fluorescence microscopy analysis has further demonstrated that this compound inhibits the generation of ROS. nih.govbiomolther.orgsemanticscholar.org Western blot analysis has indicated that this compound impacts downstream signaling molecules of ROS, including cyclooxygenase 2 (COX-2) and c-Jun N-terminal kinase (JNK). nih.gov These findings suggest that this compound inhibits ROS generation and exerts antiosteoclastogenic activity by inhibiting the COX-2 and JNK pathways. nih.gov

A TRAP (Tartrate-Resistant Acid Phosphatase) assay, used to confirm the differentiation of RAW264.7 cells into osteoclasts by LPS, showed that treatment with this compound blocked this differentiation, confirming its inhibitory effect on osteoclast differentiation. nih.govsemanticscholar.org The increased level of ROS is known to promote osteoclastogenesis and plays a significant role in the signaling involved in osteoclast activation, leading to bone resorption. nih.govbiomolther.orgsemanticscholar.org The evidence suggests that this compound inhibits osteoclastogenesis by inhibiting JNK phosphorylation, ROS production, and COX-2 expression. nih.govsemanticscholar.org

While the molecular mechanisms linking oxidative stress to osteoclast differentiation are complex, therapeutic strategies aimed at preventing ROS generation may be beneficial in addressing diseases like osteoporosis. nih.govbiomolther.org These results suggest that this compound could potentially be explored as a therapeutic agent to prevent bone resorption. nih.gov

Here is a summary of key findings regarding this compound's effect on Osteoclastogenesis:

Investigation of this compound in Alcohol Use Disorder

The potential of this compound as a pharmacotherapy for alcohol use disorder (AUD) has been investigated, driven by the need for novel and more effective treatments. researchgate.netoup.comoup.com Drug repositioning, or repurposing existing drugs, is an appealing strategy for developing new AUD therapies, potentially reducing development costs and expediting treatment availability. researchgate.netoup.com this compound, a drug with a history of clinical use for conditions like hyperuricemia and gout, has been considered for repurposing for AUD. researchgate.netoup.comoup.com

This compound inhibits organic anion transporters in the kidneys, which is relevant to its use in gout, but it also inhibits pannexin1 channels. researchgate.netoup.comoup.comwikipedia.org Pannexin1 channels are involved in purinergic neurotransmission and inflammation, processes that have been implicated in the effects of alcohol and the motivation to consume alcohol. researchgate.netoup.com

Studies in rodents have tested the effects of this compound on alcohol intake. In both alcohol-dependent and nondependent rats, as well as in mice in the drinking-in-the-dark (DID) paradigm (a model for binge-like drinking), this compound reduced alcohol intake. researchgate.netoup.comoup.com This reduction in alcohol consumption was observed without affecting the intake of water or saccharin, suggesting that the effect of this compound was selective for alcohol and not a general reduction in reward. researchgate.netoup.comoup.com Specifically, this compound dose-dependently decreased alcohol drinking in rats, with a significant effect noted at a dose of 100 mg/kg compared to a vehicle control. oup.com

These preclinical results suggest the possibility that pannexin1 could be a novel therapeutic target for the treatment of AUD. researchgate.netoup.comoup.com Given the established clinical use and safety profile of this compound, these findings support its potential as a candidate for drug repositioning for AUD. researchgate.netoup.comoup.com

Translational research, including randomized placebo-controlled alcohol interaction trials, has also explored this compound's effects in humans. One study aimed to determine if this compound reduces the pleasurable effects of alcohol drinking and the desire to drink alcohol in individuals who consume unhealthy amounts of alcohol. clinicaltrials.gov While this study was not a treatment study for AUD, it investigated this compound's effects as an investigational drug for this potential application. clinicaltrials.gov

In human trials, this compound has been reported to significantly decrease alcohol craving during the ascending limb of alcohol intoxication, compared to a placebo. researchgate.net However, this compound did not show significant effects on alcohol pharmacokinetics (Cmax, Tmax, AUC) or on the stimulant or sedative effects of alcohol. researchgate.net Inflammatory biomarkers, cognitive performance following alcohol ingestion, and hemodynamics were also not significantly affected by this compound administration in this study. researchgate.net Analysis did not reveal sex-based differences in the effects of this compound compared to placebo. researchgate.net

These data from both preclinical and translational studies support the potential of this compound for AUD treatment and reinforce the idea that pannexin 1 channels may represent a novel therapeutic target for developing new pharmacotherapies for AUD. researchgate.netoup.comoup.comresearchgate.net

Here is a summary of key findings regarding this compound's investigation in Alcohol Use Disorder:

Binding Interactions with Key Molecular Targets

Probenecid is known to interact with a variety of molecular targets, influencing their function. A significant area of research focuses on its interactions with organic anion transporters (OATs), particularly OAT1 and URAT1, which are involved in the renal excretion and reabsorption of uric acid, respectively. This compound acts as a competitive inhibitor of URAT1, reducing the reabsorption of uric acid and increasing its excretion patsnap.comnih.gov. It also inhibits OAT1, which can affect the secretion of other organic acids, including certain antibiotics patsnap.com. This compound inhibits OAT1, OAT3, and OAT6 with Ki values of 6.3, 9.0, and 8.4 µM, respectively, and OAT2 with an IC50 of 0.67 µM, showing selectivity over organic cation transporters OCT1 and OCT2 (IC50s of 1,600 and 1,700 µM) caymanchem.com.

Recent studies have also explored this compound's interactions with targets related to viral infections, such as SARS-CoV-2 and RSV. Molecular docking and simulations have indicated that this compound exhibits binding affinities with proteins like AKT1, EGFR, SRC, HSP90AA1, and PPARG. Stable interactions were observed with SRC, EGFR, and HSP90AA1, suggesting their potential roles in mediating this compound's effects nih.govfrontiersin.org. Specifically, this compound has been shown to form conventional hydrogen bonds with AKT1, EGFR, SRC, HSP90AA1, and PPARG. It also forms Pi-Donor hydrogen bonds with HSP90AA1 and Pi-Sulfur bonds with HSP90AA1 and PPARG. Carbon-hydrogen bonds have been observed with EGFR and SRC, contributing to complex stability nih.gov.

Another target for this compound is the transient receptor potential (TRP) channel TRPV2, which it activates caymanchem.comebi.ac.uk. This compound also inhibits pannexin-1 channels .

Table 1 summarizes some of the key molecular targets and the types of interactions observed with this compound.

Electrostatic Potential Mapping of this compound Binding

Electrostatic potential maps (MEPs) are utilized to predict molecular interaction sites and characterize molecular recognition patterns. These maps visualize the electrostatic potential distribution on a molecule's surface, highlighting regions of positive, negative, and neutral potential mdpi.com. For this compound, MEP analysis can reveal the areas most likely to interact with complementary charged regions on target proteins.

Studies investigating this compound binding to proteins like SRC, EGFR, and HSP90AA1 have used electrostatic potential mapping to visualize how this compound interacts within the protein cavities nih.govresearchgate.net. The color scale on these maps typically represents the electrostatic potential distribution, with red indicating negative, blue indicating positive, and white indicating neutral potential on the protein surface researchgate.net. The binding sites of this compound are often highlighted to show how the molecule fits within these electrostatic landscapes researchgate.net.

In the context of salt formation, MEP analysis of this compound has shown that the oxygen atom of the carboxyl group is a primary hydrogen bond acceptor with a negative electrostatic potential, while the oxygen atom of the sulfonyl group exhibits a global minimum electrostatic potential mdpi.com. According to the principle of electrostatic potential complementarity, regions with the highest electrostatic potential in this compound tend to bind with regions of the lowest electrostatic potential in interacting molecules mdpi.com.

Molecular Dynamics Simulations for this compound Binding

Molecular dynamics (MD) simulations are computational techniques used to study the dynamic behavior of molecular systems over time. In the context of this compound's SAR, MD simulations provide insights into the stability of this compound-protein complexes and the nature of the interactions at an atomic level nih.govfrontiersin.org.

MD simulations have been employed to further elucidate the critical interactions between this compound and its hub targets, such as AKT1, EGFR, SRC, HSP90AA1, and PPARG nih.govfrontiersin.org. Analysis of root mean square deviation (RMSD) values from these simulations can indicate the stability of the protein-ligand complex frontiersin.org. Lower and flatter RMSD values suggest greater stability of the binding pose over the simulation period frontiersin.org. Studies have shown relatively lower and flatter RMSD values for SRC-probenecid, HSP90AA1-probenecid, and EGFR-probenecid complexes compared to AKT1-probenecid and PPARG-probenecid, indicating more stable binding to the former targets frontiersin.org.

MD simulations also allow for the analysis of specific interactions, such as hydrogen bonds, van der Waals forces, and hydrophobic interactions, and their duration throughout the simulation researchgate.net. This provides a dynamic view of the binding interface and helps identify key amino acid residues involved in the interaction researchgate.net. For instance, MD simulations have confirmed that both hydrophobic interactions and hydrogen bonds play a role in the interaction between this compound and the TAS2R16 receptor tandfonline.com. The binding energies calculated from MD simulations can also provide a quantitative measure of the binding affinity tandfonline.com.

Systems Pharmacology and Bioinformatics Approaches

Systems pharmacology and bioinformatics play a significant role in understanding the multifaceted interactions of probenecid within biological systems. These approaches enable the identification of potential targets and pathways affected by this compound, moving beyond the traditional one-target, one-drug paradigm.

Studies utilizing systems pharmacology and bioinformatics have characterized targets associated with this compound for conditions such as SARS-CoV-2 and RSV co-infection frontiersin.orgresearchgate.netnih.gov. By analyzing overlapping targets between the diseases and this compound, researchers can construct protein-protein interaction (PPI) networks to visualize and understand the complex relationships frontiersin.orgresearchgate.netnih.gov. For instance, a study identified 141 overlapping targets and subsequently identified top hub targets, including AKT1, ALB, EGFR, CASP3, CTNNB1, SRC, and HSP90AA1, in the context of SARS-CoV-2/RSV co-infection frontiersin.orgresearchgate.netnih.govnih.gov.

Enrichment analysis, a bioinformatics technique, further helps in understanding the biological functions and pathways potentially modulated by this compound. These analyses suggest that this compound may influence inflammation, immunity, oxidative stress, and virus defenses, involving pathways such as Toll-like receptor, TNF, IL-17, and cytokine-cytokine receptor signaling frontiersin.orgresearchgate.netnih.govnih.govresearchgate.net.

This integrated approach provides valuable preliminary insights into the molecular mechanisms of this compound and establishes a foundation for further experimental validation frontiersin.orgresearchgate.netnih.govnih.gov.

Molecular Docking and Molecular Dynamics Simulations

Molecular docking and molecular dynamics (MD) simulations are powerful computational tools used to investigate the binding interactions between this compound and its potential target proteins at an atomic level. These methods provide insights into binding affinities, stability of complexes, and the key residues involved in the interaction.

Molecular docking analysis has been used to predict the binding poses and affinities of this compound with identified protein targets. For example, in studies investigating this compound's effects in SARS-CoV-2/RSV co-infection, molecular docking revealed strong binding affinities with targets such as AKT1, EGFR, SRC, HSP90AA1, and PPARG frontiersin.orgnih.gov. The docking scores and the formation of hydrogen bonds and other interactions provide quantitative data on the potential binding strength and nature of the interaction frontiersin.orgnih.gov.

MD simulations further validate the stability of the complexes formed between this compound and its targets over time. Analysis of parameters like Root Mean Square Deviation (RMSD) from MD simulations can indicate the stability of the binding. Studies have shown relatively lower and flatter RMSD values for complexes like SRC-probenecid, HSP90AA1-probenecid, and EGFR-probenecid, suggesting greater stability compared to others like AKT1-probenecid and PPARG-probenecid frontiersin.org. These simulations help in speculating on the likelihood of certain proteins being key targets of this compound frontiersin.orgresearchgate.netnih.gov.

Molecular modeling techniques, including docking, have also been applied to study this compound-derived compounds, such as a 1,3,4-oxadiazole-phthalimide hybrid, to explore their potential as inhibitors of enzymes like α-amylase mdpi.com. Docking scores and predicted interactions with active site residues provide evidence for the inhibitory potential of such derivatives mdpi.com.

In Vitro and In Vivo Research Models

In vitro (cell-based) and in vivo (animal) models are fundamental to experimentally validate the effects of this compound identified through computational methods and to investigate its biological activities in relevant biological contexts.

In vitro studies using various cell lines have demonstrated the direct effects of this compound on viral replication and cellular processes. For instance, this compound has shown potent inhibition of SARS-CoV-2 replication in Vero E6 cells and normal human bronchial epithelial (NHBE) cells at submicromolar concentrations mdpi.comnews-medical.netmdpi.comnih.gov. It has also shown efficacy against influenza virus in human airway epithelial cells and against human metapneumovirus (HMPV) in LLC-MK2 cells mdpi.comnews-medical.netmdpi.com. These studies often involve measuring viral titers or plaque formation to quantify the antiviral effect news-medical.net.

Beyond antiviral effects, in vitro studies have explored this compound's impact on other cellular functions. In the context of cardiovascular research, in vitro force generation studies on cardiomyocytes isolated from murine tissue exposed to this compound have shown increased myofilament force generation and calcium sensitivity nih.govahajournals.orgahajournals.org. This compound has also been shown to inhibit hypoxia-induced endothelial cell growth rate in human retinal endothelial cells frontiersin.org.

In vivo studies using animal models, such as mice and hamsters, are crucial for evaluating the efficacy of this compound in a complex biological system and assessing its effects on disease progression. Mouse models of influenza infection have shown that this compound treatment can substantially inhibit viral lung titers and protect from lethal challenge mdpi.commdpi.com. In a Syrian hamster model of SARS-CoV-2 infection, this compound administration significantly reduced viral loads in tissues news-medical.netmdpi.com.

Animal models have also been used to study this compound's effects on cardiac function and retinal angiogenesis. Translational studies in healthy and infarcted mice confirmed that this compound can improve cardiac function in a dose-dependent manner nih.gov. In a murine oxygen-induced retinopathy (OIR) model, this compound administration inhibited the upregulation of pro-angiogenic factors like VEGF and HIF-1α and ameliorated neovascularization frontiersin.org.

These in vitro and in vivo studies provide experimental evidence supporting the potential therapeutic applications of this compound identified through other research methodologies.

Translational Studies and Clinical Trial Design

Translational studies bridge the gap between basic research findings and clinical applications, while robust clinical trial design is essential for evaluating the safety and efficacy of this compound in humans.

Translational studies involving this compound have focused on translating findings from in vitro and in vivo models to human health. For example, the observation that this compound acts as an agonist of TRPV2 channels in animal models has led to its investigation for improving cardiac function in patients with heart failure with reduced ejection fraction (HFrEF) nih.govahajournals.orgahajournals.orgresearchgate.net.

Clinical trial design for this compound studies varies depending on the condition being investigated and the phase of development. Randomized, double-blind, placebo-controlled designs are commonly employed to minimize bias and provide strong evidence of efficacy.

A Phase II clinical trial investigating this compound in non-hospitalized patients with mild to moderate COVID-19 utilized a randomized, single-blind, placebo-controlled design frontiersin.orgmdpi.commdpi.commdpi.commdpi.com. This study evaluated the antiviral activity, clinical efficacy, and safety of different this compound doses mdpi.com. The design involved randomizing patients to receive either this compound or placebo and assessing outcomes such as time to viral clearance and symptom resolution mdpi.commdpi.com.

For cardiovascular applications, a study protocol for a randomized placebo-controlled clinical trial (Re-Prosper-HF) has been designed to test the hypothesis that this compound can improve cardiac function in patients with HFrEF researchgate.netx-mol.net. This study is a multi-site, double-blind, randomized-controlled trial planning to enroll a significant number of patients to assess the effects of this compound on systolic function, functional status, and self-reported health status over a defined period researchgate.net.

These examples highlight the importance of well-designed clinical trials to translate the potential benefits of this compound observed in preclinical studies into clinical practice.

Drug Repurposing Strategies for this compound

Drug repurposing, also known as drug repositioning, is a strategy that explores new therapeutic uses for existing approved or investigational drugs. This compound, with its long history of clinical use for conditions like gout, is a prime candidate for repurposing due to its known safety profile. spandidos-publications.comnih.gov

Bioinformatics tools and databases, such as DTome, facilitate drug repurposing by analyzing drug-protein interactions and secondary protein-protein interactions spandidos-publications.com. By exploring the known targets of this compound, researchers can identify potential new indications. For instance, the interaction of this compound with organic anion transporters (OATs), particularly OAT1 and OAT3, has been explored as an avenue for repurposing, especially in the context of enhancing the efficacy of antivirals and antimicrobials by blocking their renal reabsorption spandidos-publications.com.

The anti-inflammatory properties of this compound and its modulation of pathways like purinergic-pannexin-1 signaling and TRPV2 channels have led to its investigation for repurposing in cardiovascular diseases, including ischemic heart disease and heart failure nih.gov.

Recent research has focused on repurposing this compound as an antiviral agent. Studies have demonstrated its potent inhibitory effects against a range of viruses, including SARS-CoV-2, influenza virus, and respiratory syncytial virus (RSV), both in vitro and in vivo news-medical.netmdpi.comnih.govmdpi.com. This broad-spectrum antiviral activity makes this compound a promising candidate for repurposing in the treatment of various viral infections nih.gov. The potential mechanisms behind its antiviral effects, such as the modulation of ACE2 and PANX1 pathways and inhibition of OAT3, are being actively investigated frontiersin.orgmdpi.com.

Drug repurposing strategies for this compound leverage existing knowledge about its pharmacological properties and safety to accelerate the identification and development of new therapeutic applications.

Long-term Safety and Efficacy Studies in Novel Therapeutic Applications

While probenecid has been in clinical use for over 50 years with a generally limited adverse effect profile in its traditional uses, its application in novel therapeutic areas necessitates dedicated long-term safety and efficacy studies. ahajournals.org For instance, initial clinical data have demonstrated that this compound exhibits inotropic properties in patients with heart failure with reduced ejection fraction and was well tolerated in the short term. ahajournals.org Future studies are planned to focus on its long-term effects in this patient population, exploring its potential as a novel therapeutic option. ahajournals.org Translational studies in animal models have also shown that this compound can improve cardiac function parameters like inotropy and lusitropy in a dose-dependent manner without inducing malignant arrhythmias or apoptosis. ahajournals.org

Exploration of this compound's Role in Specific Disease Pathophysiologies

Emerging research is actively investigating the involvement of this compound in the pathophysiology of various diseases, leveraging its interactions with specific membrane proteins and channels. This compound has gained attention for its ability to interact with transient receptor potential vanilloid 2 (TRPV2) channels, organic anion transporters (OATs), and pannexin 1 (Panx1) hemichannels. mdpi.comnih.gov These interactions suggest potential therapeutic utility in diverse medical fields.

One significant area of exploration is the central nervous system (CNS). Studies suggest that this compound possesses neuroprotective, antiepileptic, and anti-inflammatory properties, as evidenced by its effects against neurological and neurodegenerative diseases. mdpi.com Its use as a Panx1 hemichannel blocker to reduce neuroinflammation is particularly highlighted, given neuroinflammation's role in various CNS alterations. mdpi.com this compound may also be useful in improving conventional therapies in psychiatry by increasing the bioavailability of drugs that have low passage through the blood-brain barrier or high efflux from the CNS. mdpi.com

Beyond the CNS, this compound's interaction with OATs is being investigated for novel uses. spandidos-publications.com OATs play a vital role in the distribution and excretion of many drugs. spandidos-publications.com this compound's pronounced effects on OATs suggest potential off-target effects and new therapeutic applications, including in the context of cancer development, where SLC22 family proteins (OAT1, OAT3, OAT4, and URAT1), which interact with this compound, are implicated. spandidos-publications.com

Furthermore, this compound shows potential as an antiviral agent. spandidos-publications.com It can enhance the efficacy and tolerability of other antivirals by blocking theorized viral factor uptake by OAT3. spandidos-publications.com Its interaction with multidrug-resistance protein 1 (MRP1), localized in microglia and contributing to the efflux of antivirals from the CSF, presents another avenue for repositioning this compound as an antiviral drug. spandidos-publications.com

In the cardiovascular system, this compound's activation of TRPV2 channels in cardiomyocytes is being explored for its positive inotropic and lusitropic effects, suggesting a potential role in treating heart failure. ahajournals.orgnih.gov

Development of this compound Analogs and Derivatives with Enhanced Specificity

The exploration of this compound's diverse pharmacological activities, particularly its interactions with specific transporters and channels, is expected to drive the development of novel analogs and derivatives. The aim is to create compounds with enhanced specificity for particular targets, such as specific OAT subtypes, Panx1 hemichannels, or TRPV2 channels. This targeted approach could potentially lead to therapeutic agents with improved efficacy and reduced off-target effects compared to the parent compound. Research into the structure-activity relationships of this compound and its interactions with these various proteins is crucial for the rational design of such next-generation compounds.

Refinement of Predictive Models for this compound's Pharmacological Effects

Predictive modeling plays an increasingly important role in understanding and forecasting the pharmacological effects of drugs. For this compound, refinement of predictive models is essential to better understand its complex interactions with various transporters and channels, its pharmacokinetics across different physiological states and in combination with other drugs, and its potential therapeutic outcomes in novel applications. These models can help to predict drug-drug interactions, optimize dosing strategies in new indications, and identify patient populations most likely to benefit from this compound therapy. Modeling efforts are likely to incorporate data from in vitro studies on transporter and channel interactions, in vivo pharmacokinetic data, and clinical trial results to build more accurate and comprehensive predictive frameworks.

Given the nature of the content derived from the search results, which primarily discusses ongoing research directions and potential applications, there is no structured quantitative data suitable for generating interactive data tables in this section.

Q. What mechanisms underlie this compound's anti-inflammatory effects beyond urate-lowering, and how can they be experimentally validated?

• Answer : Investigate ROS scavenging in macrophage lineages (e.g., THP-1 cells) via flow cytometry. Use siRNA knockdown of JNK/COX-2 to confirm pathway necessity. In vivo models (e.g., murine gout) can correlate ROS inhibition with reduced joint inflammation .

Methodological Notes

• Data Contradictions : Retrospective studies may overestimate efficacy due to selection bias (e.g., exclusion of non-adherent patients). Address via propensity score matching .
• Advanced Techniques : Compartmental modeling outperforms non-compartmental analysis (NCA) in predicting dose-dependent interactions .
• Clinical Relevance : this compound remains viable in mild-moderate CKD but requires SU monitoring due to variable renal clearance .