Amiloride

Time-Action Profiles in Experimental Systems

The time-action profile of this compound has been characterized in experimental systems, primarily in the context of its diuretic effects. Following oral administration, the onset of diuretic action typically occurs within 1 to 2 hours hres.cahres.canih.gov. The effect on electrolyte excretion, specifically natriuresis and potassium retention, reaches a peak between 6 and 10 hours after administration hres.cahres.ca. The duration of the diuretic effect generally lasts for approximately 24 hours hres.camims.comnih.govwikipedia.org.

In studies evaluating intranasal administration, this compound has been detected in plasma rapidly, within 10 minutes eventscribe.netresearchgate.net. The plasma concentration profile can be biphasic, with an initial peak occurring shortly after administration (within 10 minutes) followed by a second peak several hours later (between 4-8 hours) researchgate.netresearchgate.net.

Dose-Response Relationships in Preclinical Models

Dose-response relationships for this compound have been established in various preclinical models, particularly concerning its effects on electrolyte excretion. In animal studies, a dose-response relationship has been demonstrated for the actions of this compound in reducing the fractional excretion of magnesium and potassium during furosemide-induced diuresis magnesium-ges.de. While this compound shows magnesium-sparing properties, its effects on magnesium excretion are less pronounced than those on potassium excretion magnesium-ges.de.

In rats, increasing the oral dose of this compound hydrochloride from 0.25 to 4.0 mg/kg results in only moderate increases in natriuresis, although this activity persists for over 6 hours hres.cahres.ca. When administered with other diuretics like chlorothiazide, hydrochlorothiazide, or acetazolamide in rats, this compound hydrochloride increases sodium excretion and antagonizes the kaliuretic effect of the other diuretic hres.cahres.ca. Studies in dogs using oral doses of 0.1 to 0.5 mg/kg of this compound hydrochloride increased sodium excretion and decreased potassium excretion when given with ethacrynic acid or hydrochlorothiazide hres.ca.

Research in multiple myeloma cell lines has shown that this compound induces significant apoptosis in a dose-dependent manner aacrjournals.org. For example, studies using H929, KMS12-BM, and JJN3 cell lines demonstrated increased apoptosis after 24 and 48 hours of incubation with increasing concentrations of this compound (0.1–1.0 mmol/L) aacrjournals.org.

In animal models of hypertension, doses of this compound ≤1 μmol/L have been shown to mitigate cardiovascular and renal disease progression oup.com. These effects occurred without changes in steady-state serum potassium levels oup.com. This compound inhibits ENaC much more efficiently than other ion channels at submicromolar concentrations (≤1 μmol/L), with an IC50 in the range of 0.1 to 0.5 μmol/L for ENaC oup.comwikipedia.org. Higher concentrations may affect other transporters like the Na+/H+ exchanger (NHE) and Na+/Ca2+ exchanger (NCX) oup.comwikipedia.orglatoxan.com.

Interactive Data Table Placeholder: Dose-Response on Electrolyte Excretion in Rats

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Interactive Data Table Placeholder: this compound Effect on Multiple Myeloma Cell Apoptosis (Example Data)

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Studies in Liddle Syndrome Models

Liddle syndrome is a genetic disorder characterized by hypertension, hypokalemia, and metabolic alkalosis, resulting from increased activity of the epithelial sodium channel (ENaC) in the distal nephron. researchgate.netdovepress.comnih.govtandfonline.comnih.gov Mutations in the genes encoding ENaC subunits lead to increased channel activity, often by impairing their degradation or increasing their open probability. researchgate.netdovepress.comnih.govnih.gov

Preclinical studies utilizing animal models of Liddle syndrome have demonstrated that this compound, as an ENaC blocker, can effectively reduce ENaC activity. researchgate.netdovepress.com In these models, this compound, often combined with a low-sodium diet, has been shown to help restore normotension and correct electrolyte imbalances. statpearls.comresearchgate.netdovepress.com Research using whole-cell patch-clamp recordings in microdissected nephron fragments from mice with Liddle syndrome mutations indicated that the primary site of increased Na+ reabsorption is the DCT2/CNT. ahajournals.orgnih.gov These studies measured this compound-sensitive ENaC currents and found enhanced ENaC activity, particularly in the DCT2/CNT, which contributes to sodium retention and hypertension observed in Liddle syndrome models. ahajournals.orgnih.gov Furthermore, studies have shown that in Liddle syndrome, the aldosterone responsiveness of ENaC is increased, rather than decreased. ahajournals.orgnih.gov In vitro studies using Chinese Hamster Ovary (CHO) cells expressing mutant ENaC subunits from Liddle syndrome patients have also shown higher this compound-sensitive currents compared to wild-type channels, confirming the enhanced ENaC activity and its inhibition by this compound. frontiersin.org

Research in Nephrotic Syndrome Models

Nephrotic syndrome is characterized by significant proteinuria, hypoalbuminemia, edema, and hyperlipidemia. Edema formation in nephrotic syndrome is often linked to avid sodium retention in the kidneys. Preclinical studies in animal models of nephrotic syndrome have indicated that the epithelial sodium channel (ENaC) in the distal tubule is a site of sodium retention. emjreviews.comturkjnephrol.org Treatment with this compound, an ENaC blocker, has been shown to prevent volume retention in nephrotic rats and mice. emjreviews.comturkjnephrol.org This effect appears to be independent of aldosterone activity in experimental nephrotic syndrome. emjreviews.comturkjnephrol.org

Studies suggest that proteolytic activation of ENaC by proteases in the urine, resulting from aberrant glomerular filtration of plasminogen and its conversion to plasmin, may contribute to this compound-sensitive sodium retention in nephrotic syndrome. emjreviews.comturkjnephrol.orgfrontiersin.org Research in rats with induced nephrotic syndrome found higher levels of urinary plasminogen activator, and this compound treatment reduced this activity and sodium retention without altering proteinuria. frontiersin.org In animal models, this compound successfully abolishes abnormally high sodium reabsorption from the cortical collecting duct, independent of aldosterone activity. frontiersin.org

Investigations in Renal Tubular Acidosis Models

Renal Tubular Acidosis (RTA) is a group of disorders where the kidneys fail to excrete hydrogen ions or reabsorb bicarbonate properly, leading to metabolic acidosis. msdmanuals.com While the primary treatment for RTA involves alkali therapy, preclinical observations and case reports have explored the role of this compound in specific contexts. A case report described the use of this compound for the treatment of severe refractory symptomatic hypokalemia associated with type 1 renal tubular acidosis. nih.govstatpearls.com This suggests a potential role for this compound in managing electrolyte imbalances in certain forms of RTA, likely due to its potassium-sparing effects via ENaC inhibition, which reduces potassium secretion. nih.govmdpi.com

Preclinical studies have also investigated this compound's effects in models of lithium-induced kidney damage, which can lead to nephrogenic diabetes insipidus and interstitial fibrosis. otago.ac.nz In an established model of long-term lithium-induced interstitial fibrosis in rats, delayed co-administration with this compound reduced the progression of fibrosis. otago.ac.nz Further research in this model suggested that this compound attenuates pro-inflammatory pathways induced by lithium, including NF-κB and activated Akt, and increases anti-inflammatory pathways. otago.ac.nz This indicates a potential anti-inflammatory and anti-fibrotic effect of this compound in the context of lithium-induced renal injury, which may be relevant to tubular disorders.

Modulation of Renal Uric Acid Excretion

Uric acid homeostasis is maintained by a balance between production and excretion, primarily by the kidneys. Renal handling of uric acid involves filtration, reabsorption, and secretion in the proximal tubule, mediated by various urate transporters. nih.govmdpi.comresearchgate.net

Prolonged use of this compound can reduce the excretion of renal uric acid. nih.govstatpearls.com This effect is thought to be related to volume contraction caused by its diuretic action, which can lead to increased reabsorption of uric acid from the proximal convoluted tubule. nih.govstatpearls.com Some studies suggest that the transport activity of certain organic anion transporters (OATs), like OAT4, which are involved in uric acid transport, may be coupled with the effect of NHE3. nih.gov this compound is known to inhibit NHE3 at higher concentrations, which could potentially influence the activity of transporters like OAT4, indirectly affecting uric acid handling. However, the primary mechanism for reduced uric acid excretion with prolonged this compound use appears to be related to volume status. nih.govstatpearls.com

Antihypertensive Mechanisms Beyond Diuresis

While this compound's diuretic action contributes to its antihypertensive effect, preclinical studies suggest additional mechanisms. This compound has the potential to cause vasodilation. nih.govstatpearls.com

Furthermore, the epithelial sodium channel (ENaC) is present and functional in non-renal epithelial cells, including those in the cardiovascular system such as endothelial cells, smooth muscle cells, and fibroblasts. oup.com Preclinical studies have shown that this compound can antagonize or prevent the actions of aldosterone in these non-renal tissues in animal models of salt-dependent hypertension. oup.com This suggests that ENaC inhibition in the cardiovascular system might contribute to this compound's antihypertensive effects, independent of its renal actions.

Research in experimental hypertension animal models has indicated that this compound can mitigate cardiovascular and renal disease progression even in the presence of elevated blood pressure. oup.com These benefits have been observed without significant changes in steady-state serum potassium levels in some studies. oup.com this compound's ability to prevent or mitigate cardiac fibrosis in experimental models has also been noted, with the postulated mechanism potentially related to the maintenance of myocardial potassium concentration. oup.com

Preclinical models also indicate that this compound, as a nonselective antagonist of acid-sensing ion channels (ASICs), may influence baroreflex sensitivity and blood pressure regulation. physiology.orgnih.gov However, studies translating these findings to humans have shown contrasting results, with a single oral dose of this compound not affecting spontaneous arterial baroreflex sensitivity and blood pressure variability in healthy young adults. physiology.orgnih.gov

In addition to direct vascular effects and non-renal ENaC inhibition, preclinical research suggests this compound may have anti-inflammatory effects relevant to cardiovascular health. Studies in angiotensin II-hypertensive mice with type 1 diabetes treated with this compound showed increased TNF in cardiac and kidney tissues. physiology.org However, in vitro studies using lipopolysaccharide-stimulated human macrophages demonstrated that this compound and benzamil (an this compound analog) decreased pro-inflammatory cytokines like TNF, IL-6, IL-10, and IL-1β. physiology.org This suggests a potential direct action of this compound on macrophages to reduce pro-inflammatory cytokine release, which could contribute to its cardiovascular benefits. physiology.org

Cardioprotective Effects

Preclinical investigations have explored the potential cardioprotective effects of this compound, particularly in the context of myocardial ischemia-reperfusion injury and cardiac remodeling. This compound, as an inhibitor of the Na+/H+ exchanger (NHE1), has shown promise in reducing intracellular sodium and consequently calcium accumulation, which are key factors in myocardial injury during ischemia-reperfusion. mdpi.comjacc.org Studies in isolated rat hearts subjected to ischemia/reperfusion demonstrated that this compound treatment significantly decreased the release of lactate dehydrogenase (LDH) and creatine kinase (CK), markers of cardiac damage, and reduced infarct size. nih.gov This suggests a protective effect against ischemia/reperfusion-induced injury.

Furthermore, research in animal models of hypertension and obesity has indicated that this compound may mitigate cardiovascular and renal disease progression and reduce vascular stiffness. oup.comresearchgate.net Inhibition of ENaC with this compound has been shown to decrease oxidative stress, endothelium permeability, inflammation, arterial fibrosis, and aortic stiffness in obese mice. researchgate.net These effects were observed even in the absence of changes in blood pressure or sodium retention in some studies. researchgate.net

However, the translation of these positive preclinical findings with NHE1 inhibitors, including this compound, into successful clinical outcomes for ischemic heart disease has faced challenges. mdpi.comjacc.org

Cystic Fibrosis Research

This compound has been extensively investigated for its potential therapeutic use in Cystic Fibrosis (CF), a genetic disorder characterized by abnormal airway epithelial electrolyte transport leading to dehydrated airway surface liquid (ASL) and impaired mucociliary clearance. ersnet.orgatsjournals.organnualreviews.org The rationale for using this compound in CF is based on its ability to block the epithelial sodium channel (ENaC), which is hyperactive in CF airways, leading to excessive sodium and water absorption. ersnet.orgatsjournals.orgnih.govatsjournals.org By inhibiting ENaC, this compound is hypothesized to reduce excessive sodium and water absorption, thereby improving the hydration of the ASL and enhancing mucus clearance. ersnet.orgatsjournals.orgresearchgate.net

Studies have shown that application of this compound to CF airway epithelia in vitro reduces sodium absorption. ersnet.org In patients with CF, inhalation of a single dose of this compound has been shown to increase mucociliary clearance. ersnet.org

Despite the promising theoretical basis and initial positive findings, clinical trials with nebulized this compound in CF patients have yielded mixed results. Some early studies suggested a potential benefit, including a reduction in the rate of decline in forced vital capacity (FVC) and improvement in sputum rheological properties ersnet.orgnih.gov. However, other studies, including a randomized, double-blind, placebo-controlled, cross-over trial, did not find significant additional clinical benefit from nebulized this compound in patients receiving established CF treatment. ersnet.orgnih.gov

Data from a study comparing nebulized this compound and placebo over two six-month periods in CF patients indicated no significant changes in several parameters, as shown in the table below. ersnet.org

The limited clinical efficacy of this compound in CF may be attributed to factors such as its relatively low potency and short duration of action on the airway surface, as well as rapid absorption from the mucosal surface. atsjournals.orgscilit.com

Impact on Airway Surface Liquid Hydration

In CF, increased sodium absorption across airway epithelia leads to increased osmotic water absorption, which is thought to reduce the hydration of the airway surface liquid (ASL). ersnet.orgresearchgate.net this compound, by blocking ENaC, is postulated to increase ASL volume by inhibiting sodium and fluid absorption. researchgate.net In vitro studies and research using animal models of CF lung disease have shown that inhibition of ENaC can restore ASL hydration. researchgate.net

However, studies investigating the direct impact of this compound inhalation on sputum water content in CF patients have not always shown a measurable change, suggesting that the mechanism for mucus improvement by this compound might be more complex than solely increasing bulk water content. atsjournals.org Despite this, the rationale remains that improving ASL hydration is a key therapeutic target in CF, and this compound's ability to block ENaC aligns with this goal. annualreviews.orgresearchgate.net

Effects on Mucus Rheology and Clearance

Abnormal mucus rheology and impaired mucociliary clearance are central features of CF lung disease. atsjournals.orgatsjournals.orgnih.gov The dehydration of the ASL in CF contributes to thickened mucus that is difficult to clear by ciliary action and coughing. annualreviews.orgnih.govatsjournals.org

This compound inhalation therapy has been hypothesized to improve the rheological properties of mucus, making it more amenable to clearance. atsjournals.org Some studies have reported that this compound treatment improved measured indexes of sputum viscosity and elasticity and enhanced calculated indexes of mucociliary and cough clearance in adult CF patients. nih.gov One study indicated that chronic this compound therapy increased sputum sodium and chloride content and decreased the principal index of sputum rigidity (log G*). atsjournals.org

Data from a study on this compound inhalation therapy in CF patients showed changes in sputum ion content and rheology: atsjournals.org

While this compound can influence the bioelectric properties of the respiratory mucosa, its direct effect on human bronchial ciliary activity in vitro appears to be weak and of short duration, suggesting it may not play a major role in mucociliary transport through this mechanism. karger.com

Interaction with Hypertonic Saline Therapy

Hypertonic saline (HS) inhalation is a widely used therapy in CF that works by creating an osmotic gradient to draw water onto the airway surface, thereby increasing ASL volume and improving mucus clearance. annualreviews.orgnih.govphysiology.org Interestingly, research has explored the interaction between this compound and hypertonic saline therapy in CF.

Some studies have shown that this compound can diminish the beneficial effects of hypertonic saline on sustained mucociliary clearance and pulmonary function in vivo. annualreviews.orgresearchgate.net This paradoxical observation suggests a complex interaction between the mechanisms of action of these two therapies. One proposed explanation is that this compound might inhibit aquaporin water channels, thereby negating the osmotic effect of hypertonic saline. researchgate.net However, other studies have argued against the inhibition of water channels as the primary mechanism for this compound's effect on hypertonic saline response. physiology.org

In vitro studies on human bronchial epithelial cells from CF donors have shown that exposure to hypertonic saline inhibits sodium current (INa), and the inclusion of this compound with HS led to an increased recovery of INa, paradoxically protecting the epithelium from the inhibitory effects of HS. physiology.orgphysiology.org This further highlights the intricate interplay between this compound and hypertonic saline at the cellular level.

Studies comparing hypertonic saline alone versus hypertonic saline with this compound have indicated that hypertonic saline alone was more effective in increasing mucociliary clearance. nih.gov

Acute Respiratory Distress Syndrome (ARDS) Research

Preclinical studies have investigated the potential of this compound in the context of Acute Respiratory Distress Syndrome (ARDS). ARDS is characterized by diffuse alveolar injury, including excessive inflammation, increased epithelial and vascular permeability, and alveolar edema. nih.gov While there is significant research into the pathogenesis of ARDS, effective pharmacotherapies remain limited, and treatment is primarily supportive. nih.gov

Research has explored the role of epithelial sodium channels (ENaC) in alveolar fluid clearance, a process often impaired in ARDS. This compound is known to inhibit ENaC. researchgate.netthieme-connect.com Studies involving mesenchymal stromal cells (MSCs) and their conditioned medium (CM) have shown therapeutic potential in ARDS models. researchgate.net The beneficial effects of MSC-CM on alveolar fluid clearance in endotoxin-injured lung models were significantly reduced when this compound was added to the medium, suggesting that the therapeutic effect is, in part, mediated by mechanisms sensitive to ENaC inhibition. researchgate.net Specifically, the addition of this compound (5 x 10-4 M) to MSC-CM reduced the therapeutic effect on alveolar fluid clearance by 56% in an ex vivo perfused lung model. researchgate.net This indicates that this compound's interaction with ENaC may influence the pathways involved in resolving pulmonary edema in ARDS.

Neuroprotective Effects in Multiple Sclerosis Models

Neurodegeneration is a significant contributor to permanent disability in multiple sclerosis (MS). nih.govoup.com Current treatments primarily target inflammation but have shown less efficacy in preventing long-term disability. nih.govoup.com Targeting ASIC1, which is over-expressed in acute MS lesions and contributes to injurious intracellular ion accumulation, appears to be a potential strategy for limiting cellular injury and neurodegeneration. nih.govoup.com

This compound, as an ASIC1 inhibitor, has demonstrated neuroprotective and myeloprotective effects in experimental models of multiple sclerosis. nih.govoup.com These effects appear to occur independently of any significant anti-inflammatory influence. oup.com A translational study tested the neuroprotective effects of this compound in patients with primary progressive MS. nih.govoup.com This study assessed ASIC1 expression in chronic brain lesions and evaluated this compound's effect using MRI markers of neurodegeneration. nih.govoup.com Increased expression of ASIC1 was found in axons within chronic inactive lesions of progressive MS patients. nih.govoup.com Patients treated with this compound showed a significant reduction in the normalized annual rate of whole-brain volume loss compared to the pretreatment phase. nih.govoup.com Changes in diffusion indices of tissue damage in white and deep grey matter structures were also significantly reduced during the treatment phase. nih.govoup.com

This compound was also one of three drugs studied in the MS-SMART trial involving individuals with secondary progressive MS, although that trial did not find a clinical effect of the tested drugs. mstrust.org.ukmscanada.ca

Acid-Sensing Ion Channel (ASIC) Inhibition in Panic Disorder Research

Anxiety and panic disorders are common mental health conditions with a need for more effective treatments. ukri.orgnih.govresearchgate.netuoa.gr Acid-sensing ion channels (ASICs) in the brain have been associated with fear conditioning and anxiety responses, suggesting they could be therapeutic targets for panic disorder. ukri.orgnih.govresearchgate.netuoa.gr

This compound is an inhibitor of ASICs in the brain and has been shown to reduce panic symptoms in preclinical animal models. nih.govresearchgate.netuoa.grresearchgate.net Research suggests that ASIC inhibitors should have therapeutic potential in human anxiety. ukri.org Studies have explored the pharmacokinetics of intranasal this compound as a potential rapid-onset treatment for acute panic attacks. nih.govresearchgate.netuoa.grresearchgate.net Preclinical studies have shown that inhalation of this compound prevents panic symptoms in mice exposed to anxiogenic stimuli. researchgate.net

Anti-Myeloma Activity

This compound has demonstrated potent anti-myeloma activity in preclinical studies using myeloma cell lines and xenograft mouse models. aacrjournals.orgresearchgate.netaacrjournals.orgrarecancernews.comnih.govnih.govhealthtree.org Its activity has been observed in a broad panel of myeloma cell lines, regardless of TP53 status. researchgate.netnih.gov this compound also showed a synergistic effect when combined with other anti-myeloma drugs such as dexamethasone, melphalan, lenalidomide, and pomalidomide. aacrjournals.orgresearchgate.netaacrjournals.orgrarecancernews.comnih.govhealthtree.org

Beyond directly inducing cytotoxicity, this compound has been found to significantly alter the level of transcript isoforms and alternative splicing events, and deregulate the spliceosomal machinery in myeloma cells. aacrjournals.orgaacrjournals.orgrarecancernews.comnih.gov Disruption of the splicing machinery was associated with the inhibition of myeloma cell viability after this compound exposure. aacrjournals.orgaacrjournals.orgnih.gov

Induction of Apoptosis (Caspase-dependent and independent mechanisms)

A key mechanism by which this compound exerts its anti-myeloma activity is the induction of apoptosis. aacrjournals.orgresearchgate.netaacrjournals.orgnih.govnih.gov This apoptosis induction has been observed in various myeloma cell lines in a dose- and time-dependent manner. aacrjournals.orgresearchgate.net

Research indicates that the apoptosis induced by this compound in multiple myeloma cells is mediated by both caspase-dependent and caspase-independent mechanisms. aacrjournals.orgresearchgate.net Studies have shown significant activation of caspases 3/7, 8, and 9 in tested myeloma cell lines following this compound treatment. aacrjournals.org The involvement of a caspase-independent mechanism was also observed, as a pan-caspase inhibitor (Z-VAD-FMK) could inhibit caspase-3/7 activity but not the apoptosis induced by this compound. aacrjournals.org This dual mechanism of action contributes to this compound's cytotoxic effects on myeloma cells.

Data Table: this compound's Effects on Myeloma Cell Lines

Note: Data compiled from search result aacrjournals.org. Specific quantitative data for all metrics across all cell lines were not consistently available in the provided snippets.

Synergistic Effects with Chemotherapeutic Agents

Preclinical studies have indicated that this compound can exhibit synergistic effects when combined with certain chemotherapeutic agents. In the context of multiple myeloma, this compound demonstrated synergistic activity when used alongside dexamethasone, melphalan, lenalidomide, and pomalidomide. aacrjournals.orgrarecancernews.comaacrjournals.org This suggests that this compound could potentially enhance the efficacy of existing treatments for this hematological malignancy. Additionally, in canine osteosarcoma cells, this compound showed strong synergism with doxorubicin, leading to improved response compared to either agent alone. nih.gov This synergistic effect was associated with the activation of p53 signaling and downregulation of Akt phosphorylation, ultimately increasing apoptosis. nih.gov

Impact on Splicing Machinery and Gene Regulation

This compound has been found to modulate alternative splicing in various human cancer cells. aacrjournals.orgrarecancernews.com Alternative splicing is a crucial process that can be dysregulated in cancer, contributing to tumor progression and metastasis. aacrjournals.orgrarecancernews.com Studies have shown that this compound can alter the level of transcript isoforms and alternative splicing events, as well as deregulate the spliceosomal machinery. aacrjournals.orgaacrjournals.org This disruption of the splicing machinery has been linked to the inhibition of cancer cell viability. aacrjournals.orgaacrjournals.org For instance, in multiple myeloma cells, this compound treatment led to the modulation of the splicing machinery, observed through changes in the immunofluorescent staining of the SR protein SC35, which correlated with reduced cell viability. aacrjournals.org this compound has also been shown to "normalize" the splicing of certain oncogenic RNA transcripts, such as BCL-X, HIPK3, and RON/MISTR1, in human hepatocellular carcinoma cells. plos.orgresearchgate.netnih.gov This modulation of splicing is thought to be associated with the hypo-phosphorylation of splicing factors like SF2/ASF. plos.orgresearchgate.netnih.gov

Role of p53 Signaling

Research indicates a role for p53 signaling in the anticancer activity of this compound, particularly in multiple myeloma. Activation of p53 signaling has been observed in multiple myeloma cells with both wild-type and mutated TP53 after exposure to this compound. aacrjournals.orgrarecancernews.comaacrjournals.orgresearchgate.net This activation appears to contribute to this compound-induced apoptosis in cells expressing p53. aacrjournals.orgresearchgate.net However, this compound has also been shown to induce apoptosis in multiple myeloma cells lacking p53 expression, suggesting that mechanisms independent of p53 signaling are also involved in its antimyeloma effect. aacrjournals.orgrarecancernews.comaacrjournals.orgresearchgate.net In canine osteosarcoma cells, combination treatment with this compound and doxorubicin significantly upregulated p53-mitochondrial signaling, leading to increased apoptosis. nih.gov

Breast Cancer Research

This compound and its derivatives have attracted attention as potential therapeutic agents for breast cancer. biorxiv.orgresearchgate.net Early studies characterized amilorides as inhibitors of sodium-proton antiporter-dependent tumor growth and urokinase plasminogen activator-mediated metastasis. biorxiv.orgbiorxiv.org More recent observations suggest that this compound derivatives can be selectively cytotoxic towards tumor cells, including those resistant to conventional therapies. biorxiv.orgresearchgate.net

Lysosome-Dependent Cell Death Mechanisms

Recent studies highlight the induction of lysosome-dependent cell death (LDCD) as a mechanism by which this compound derivatives exert their cytotoxic effects in breast cancer cells, particularly in therapy-resistant populations. biorxiv.orgresearchgate.netbiorxiv.orgmdpi.com Lipophilic this compound derivatives, such as hexamethylene this compound (HMA) and LLC1, have been shown to induce lysosomal membrane permeabilization (LMP), a critical step preceding lysosome-dependent cell death. biorxiv.orgresearchgate.netbiorxiv.orgmdpi.comescholarship.orgnih.gov This process involves the release of lysosomal contents, such as cathepsins, into the cytosol, triggering cell death. biorxiv.orgmdpi.com Cancer cell lysosomes are considered particularly susceptible to LMP, providing a rationale for targeting lysosomes in cancer therapy. mdpi.com

Dual-Targeting Strategies (e.g., uPA and NHE1)

This compound's anticancer effects have been linked to its inhibitory activity against two distinct targets: urokinase-type plasminogen activator (uPA) and sodium-hydrogen exchanger isoform-1 (NHE1). researchgate.netmdpi.comnih.gov uPA is a serine protease involved in matrix degradation and tumor cell invasiveness, while NHE1 is a key regulator of transmembrane pH that supports carcinogenic progression. researchgate.netmdpi.comnih.gov this compound exhibits moderate inhibitory activity against both targets. researchgate.netmdpi.com Research is exploring the development of this compound derivatives with enhanced potency and selectivity, including compounds with dual-targeting activity against both uPA and NHE1, as well as those selective for a single target. mdpi.comnih.gov Inhibition of uPA by this compound has been shown to suppress the invasive capacity of human breast cancer cells. nih.govnih.gov Similarly, NHE1 inhibition has been investigated as a strategy to potentiate the effects of chemotherapy in breast cancer, including increasing sensitivity to doxorubicin and paclitaxel. nih.govresearchgate.net

Modulation of Cancer Cell pH

The tumor microenvironment is often characterized by extracellular acidity, which is known to contribute to cancer progression, including migration, invasion, and resistance to therapy. dovepress.comnih.gov Cancer cells maintain a relatively neutral to slightly alkaline intracellular pH by increasing the activity of proton extrusion mechanisms, such as the Na+/H+ exchanger isoform 1 (NHE1). dovepress.comnih.gov

This compound and its derivatives are known inhibitors of NHE1. nih.govnih.gov By inhibiting NHE1, this compound can interfere with the ability of cancer cells to extrude protons, potentially leading to intracellular acidification. dovepress.com This modulation of intracellular and extracellular pH can have several effects on cancer cells. Studies have shown that interfering with pH regulating systems is considered a potential therapeutic strategy in oncology. dovepress.commdpi.com

Preclinical studies have investigated the effects of this compound on cancer cell viability and pH in various cancer types, including prostate cancer. mdpi.com For instance, in vitro studies using PC3 prostate cancer cells demonstrated that this compound treatment could alter extracellular pH in a dose-dependent manner, moving the extracellular environment towards a more neutral pH value compared to control cells. mdpi.com

Data from studies on PC3 prostate cancer cells illustrate the dose-dependent effect of this compound on extracellular pH after 24 hours of treatment:

This compound treatment has also been shown to induce a dose-dependent decrease in the viability of PC3 cells under normoxic conditions, with a more pronounced effect observed at higher concentrations and longer incubation times. mdpi.com While this compound's primary mechanism in this context is linked to NHE1 inhibition and subsequent pH modulation, it has also been shown to impede prostate cancer tumor growth by inhibiting urokinase-type plasminogen activator (uPA), an enzyme involved in cell migration and invasion. nih.gov Furthermore, this compound has demonstrated the ability to decrease the invasiveness of aggressive prostate cancer cells and reduce the size of prostate cancer xenografts in rodent models. nih.gov

Blockade of Proinflammatory Cytokine Production (e.g., IL-1β, TNF-α, IL-6)

Research indicates that this compound can suppress the production of key proinflammatory cytokines. Exposure of alveolar epithelial cells to this compound or its analog, 5-(N,N-hexamethylene)-amiloride (HMA), reduced lipopolysaccharide (LPS)-induced secretion of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in a dose-dependent manner. atsjournals.orgatsjournals.orgnih.gov This inhibitory effect was associated with the augmentation of a counter anti-inflammatory response. atsjournals.orgatsjournals.org this compound has also been shown to inhibit IL-8 and IL-6 release from respiratory syncytial virus (RSV)-infected airway epithelium. physiology.org In LPS-stimulated human THP-1 macrophages, this compound and benzamil decreased TNF, IL-6, IL-10, and IL-1β. physiology.orgresearchgate.net

The following table summarizes some findings on this compound's effect on proinflammatory cytokine production:

Amplification of Anti-inflammatory Loop Responses (e.g., IL-6, IL-10)

In addition to blocking proinflammatory cytokines, this compound has been observed to amplify anti-inflammatory responses. In alveolar epithelial cells, the inhibitory effect of this compound on proinflammatory cytokines was associated with the augmentation of a counter anti-inflammatory response mediated by IL-6 and IL-10. atsjournals.orgatsjournals.orgnih.gov This suggests a dual role for this compound in modulating the inflammatory balance. atsjournals.orgatsjournals.org

Regulation of NF-κB Pathway

The nuclear factor-kappa B (NF-κB) pathway is a critical regulator of inflammatory responses. atsjournals.orgatsjournals.orgresearchgate.netwikipedia.orgresearchgate.net Analysis of the mechanism behind this compound's immunomodulatory effects has revealed the involvement of an inhibitory kappaB (IκB-α)/NF-κB-sensitive pathway. atsjournals.orgatsjournals.orgnih.gov this compound and its analog HMA suppressed the phosphorylation of IκB-α mediated by LPS, which allowed for its cytosolic accumulation. atsjournals.orgatsjournals.orgnih.gov Furthermore, both inhibitors interfered with the nuclear translocation of selective NF-κB subunits, an effect linked to the blockade of NF-κB DNA-binding activity. atsjournals.orgatsjournals.orgnih.gov This indicates that this compound can intervene with the nuclear translocation of selective NF-κB subunits, thereby blockading NF-κB activation. atsjournals.org

Role in Macrophage-Derived Cytokine Release

Macrophages play a significant role in the inflammatory response through the release of cytokines. nih.govresearchgate.net Studies suggest that epithelial Na+ channel (ENaC) activity may contribute to macrophage-derived cytokine release. physiology.orgresearchgate.net Inhibition of ENaC by this compound has been shown to reduce proinflammatory cytokines TNF and IL-6 in patients with type 2 diabetes and in THP-1-derived macrophages in vitro, potentially through a direct action on macrophages. physiology.orgresearchgate.net In LPS-stimulated human THP-1 macrophages, this compound and benzamil decreased TNF, IL-6, IL-10, and IL-1β. physiology.orgresearchgate.net

Inhibition of Viral RNA Polymerase

Blockade of Proinflammatory Cytokine Production (e.g., IL-1β, TNF-α, IL-6)

Research indicates that this compound can reduce the production of key proinflammatory cytokines. In alveolar epithelial cells, exposure to this compound or its analog, HMA, led to a dose-dependent reduction in the secretion of LPS-induced IL-1β and TNF-α. atsjournals.orgatsjournals.orgnih.gov This inhibitory effect was observed in conjunction with the amplification of a counter anti-inflammatory response. atsjournals.orgatsjournals.org this compound has also been shown to inhibit the release of IL-8 and IL-6 from airway epithelium infected with RSV. physiology.org In studies using LPS-stimulated human THP-1 macrophages, this compound and benzamil were found to decrease levels of TNF, IL-6, IL-10, and IL-1β. physiology.orgresearchgate.net

The following table summarizes some findings on this compound's effect on proinflammatory cytokine production:

Amplification of Anti-inflammatory Loop Responses (e.g., IL-6, IL-10)

Beyond its role in blocking proinflammatory cytokines, this compound has been noted to amplify anti-inflammatory responses. In alveolar epithelial cells, the reduction in proinflammatory cytokines by this compound was linked to the increased presence of an anti-inflammatory response mediated by IL-6 and IL-10. atsjournals.orgatsjournals.orgnih.gov This observation supports a dual function for this compound in influencing the balance of inflammation. atsjournals.orgatsjournals.org

Regulation of NF-κB Pathway

The NF-κB pathway is a key regulator in inflammatory processes. atsjournals.orgatsjournals.orgresearchgate.netwikipedia.orgresearchgate.net Investigations into the mechanism underlying this compound's immunomodulatory effects have pointed to the involvement of an IκB-α/NF-κB-sensitive pathway. atsjournals.orgatsjournals.orgnih.gov this compound and its analog HMA were found to suppress the LPS-mediated phosphorylation of IκB-α, leading to its accumulation in the cytosol. atsjournals.orgatsjournals.orgnih.gov Furthermore, both inhibitors interfered with the movement of specific NF-κB subunits into the nucleus, an effect associated with blocking the DNA-binding activity of NF-κB. atsjournals.orgatsjournals.orgnih.gov These findings suggest that this compound can disrupt the nuclear translocation of specific NF-κB subunits, thereby inhibiting NF-κB activation. atsjournals.org

Role in Macrophage-Derived Cytokine Release

Macrophages are significant contributors to the inflammatory response through the release of cytokines. nih.govresearchgate.net Studies propose that the activity of the epithelial Na+ channel (ENaC) may play a role in cytokine release from macrophages. physiology.orgresearchgate.net Blocking ENaC with this compound has been shown to decrease the levels of proinflammatory cytokines TNF and IL-6 in patients with type 2 diabetes and in THP-1-derived macrophages in vitro, potentially indicating a direct impact on these cells. physiology.orgresearchgate.net In LPS-stimulated human THP-1 macrophages, this compound and benzamil reduced the levels of TNF, IL-6, IL-10, and IL-1β. physiology.orgresearchgate.net

Osteoporosis Treatment Potential

While the provided outline includes osteoporosis treatment potential as an emerging research area for this compound, the search results did not yield specific information detailing this compound's role or potential in the treatment of osteoporosis. Therefore, content for this subsection cannot be provided based on the available search data.

Drug-Resistant Tuberculosis Research

Increasing antimicrobial resistance necessitates the identification of new therapeutic agents with novel or multiple molecular targets. researchgate.netnih.gov Energy conservation in Mycobacterium tuberculosis (Mtb), the bacterium responsible for tuberculosis, has been identified as a promising target for combating drug-resistant strains. researchgate.netnih.govufl.li

Research has shown that derivatives of this compound hold potential as anti-tubercular inhibitors. researchgate.netnih.gov Specifically, a 6-substituted derivative of this compound, known as HM2-16F, has demonstrated anti-tubercular activity comparable to bedaquiline, an FDA-approved drug used for drug-resistant tuberculosis. researchgate.netnih.govufl.li HM2-16F has also shown efficacy against bedaquiline-resistant mutants of Mtb. researchgate.netnih.govufl.li

The mechanism of action of HM2-16F involves interfering with bacterial energy conservation. researchgate.net Studies indicate that HM2-16F weakly inhibits the F1Fo-ATP synthase in Mtb, leading to ATP depletion. researchgate.netnih.govufl.li Additionally, it affects the entry of acetyl-CoA into the Krebs cycle. researchgate.netnih.govufl.li HM2-16F has also been shown to synergize with Q203 (Telacebec), a cytochrome bcc-aa3 oxidase inhibitor. researchgate.netnih.govufl.li Co-administration of HM2-16F and Q203 has been shown to sterilize in vitro cultures of Mtb within 14 days, with the synergy attributed to the direct inhibition of cytochrome bd oxidase by HM2-16F. researchgate.netnih.govufl.li These findings suggest that this compound derivatives could serve as a platform for discovering new drugs targeting energy generation in drug-resistant tuberculosis. researchgate.netnih.govufl.li

Thiazolidinediones-Induced Edema

Thiazolidinediones (TZDs), a class of drugs used to treat type 2 diabetes, are known to cause fluid retention and edema as a common side effect. vumc.orgnih.gov This fluid retention is a significant clinical limitation and can be particularly concerning for patients with or at high risk for heart failure. vumc.orgnih.govcore.ac.ukescholarship.org

Increasing evidence from animal models suggests that TZD-induced fluid retention is due to increased sodium retention in the distal nephron. core.ac.uk TZDs, which are agonists of peroxisome proliferator-activated receptor gamma (PPARγ), have been shown to stimulate sodium reabsorption in the distal nephron, potentially by upregulating the expression and translocation of ENaC. nih.govescholarship.orgnih.govpnas.org PPARγ is expressed in the renal cortical collecting duct, a region involved in regulating sodium and water balance via ENaC. nih.govpnas.org

Preclinical studies in mice have demonstrated that this compound, an ENaC blocker, can prevent the fluid retention and body weight gain induced by thiazolidinediones like pioglitazone. vumc.orgnih.govescholarship.org This suggests a potential role for this compound in counteracting the sodium retention caused by TZDs. vumc.orgnih.govcore.ac.ukescholarship.org Studies in human patients with type 2 diabetes who experienced rosiglitazone-induced fluid retention also showed that this compound was effective in arresting the continued plasma volume expansion. nih.gov These results support the hypothesis that ENaC upregulation plays a role in the pathophysiology of TZD-induced fluid retention. nih.gov

However, there can be discrepancies in findings between species and specific TZD compounds. For instance, one study in rats showed that post-treatment with this compound unexpectedly exacerbated fluid retention induced by farglitazar, highlighting the complexity of these interactions. nih.gov Despite this, the evidence from multiple studies, particularly those focusing on pioglitazone and rosiglitazone, supports the potential of this compound as a therapeutic strategy to mitigate TZD-induced fluid retention by targeting ENaC-mediated sodium reabsorption in the collecting duct. vumc.orgnih.govescholarship.orgnih.govpnas.org

Lithium-Induced Polyuria

Lithium is a medication used to treat affective disorders, but one of its major side effects is nephrogenic diabetes insipidus, characterized by polyuria (excessive urination). nih.govmedcraveonline.comopenaccessjournals.comnih.gov This occurs because lithium can inhibit vasopressin-mediated water reabsorption in the renal collecting tubules by interfering with the function and expression of aquaporin 2. medcraveonline.comopenaccessjournals.comnih.gov Lithium enters the renal principal cells primarily through the epithelial sodium channel (ENaC). medcraveonline.comopenaccessjournals.compsychiatrictimes.com

This compound, by blocking ENaC, can prevent the entry of lithium into these renal cells, thereby blunting lithium's inhibitory effect on water transport and reducing polyuria. medcraveonline.comopenaccessjournals.comnih.govpsychiatrictimes.com Preclinical studies in animals demonstrated that this compound could reduce lithium-induced polyuria without affecting lithium or potassium levels. nih.gov

Studies in patients with lithium-induced polyuria have shown that this compound administration leads to a decrease in urine volume and an increase in urine osmolality, indicating improved renal concentrating ability. nih.govnih.gov This reduction in urine output has been observed without significant changes in plasma lithium, potassium, or bicarbonate levels, or urinary excretion of sodium or lithium. nih.gov The effect of this compound in mitigating lithium-induced polyuria is attributed, at least in part, to its ability to block lithium entry into the collecting tubule cells via ENaC. medcraveonline.comopenaccessjournals.comnih.govpsychiatrictimes.com

This compound is often considered a preferred treatment option for lithium-induced polyuria compared to thiazide diuretics because it is less likely to increase serum lithium concentrations to potentially toxic levels and does not contribute to hypokalemia. openaccessjournals.compsychiatrictimes.comdroracle.ai

The mechanism by which this compound counteracts lithium's nephrotoxic effects is primarily the inhibition of ENaC in the principal cells of the nephron, which prevents lithium accumulation and the subsequent downregulation of Aquaporin 2 expression. medcraveonline.com This action helps to restore the kidney's ability to concentrate urine. nih.govmedcraveonline.comnih.gov

Here is a summary of the research findings discussed:

Cell Culture Systems

Cell culture systems are widely used to study the effects of this compound on various cell lines. These include investigations into its potential therapeutic applications and its interactions with ion channels and transporters.

Myeloma Cell Lines: this compound has demonstrated potent anti-myeloma activity in a broad panel of multiple myeloma (MM) cell lines. Studies have shown that this compound induces apoptosis in these cell lines. aacrjournals.orgresearchgate.net Furthermore, this compound has exhibited synergistic effects when combined with other therapeutic agents such as dexamethasone, melphalan, lenalidomide, and pomalidomide in MM cell lines. aacrjournals.orgresearchgate.net Research using myeloma cell lines has also investigated the mechanisms of action of this compound, including its impact on gene expression and splicing. aacrjournals.orgresearchgate.net Hexamethylene this compound (HA), a derivative of this compound, has also been shown to inhibit growth and induce apoptosis in MM cell lines, including those resistant to carfilzomib. d-nb.info

Breast Cancer Cell Lines: Breast cancer cell lines, such as T47D and MCF-7, have been used to study the effects of factors from polarized THP-1 macrophages on their cellular and functional changes, including the induction of a partial epithelial-mesenchymal transition (EMT) phenotype and increased migratory and invasive properties. oncotarget.com While these studies focus on macrophage influence on breast cancer cells, this compound's known inhibitory effects on Na+/H+ exchangers (NHEs), which are implicated in cancer progression, suggest that breast cancer cell lines could also be utilized to investigate this compound's direct or indirect impact on these processes. cdnsciencepub.com

Human Bronchial Epithelial Cells: Studies on human bronchial epithelial cells utilize techniques like ion-selective electrodes to measure ion transport. mdpi.com These cells possess various channels on their apical face, including the this compound-sensitive epithelial Na+ channel (ENaC). mdpi.com

THP-1 Macrophages: The human monocytic cell line THP-1 is commonly used to generate macrophages for in vitro studies, including those investigating the tumor microenvironment and the interaction between macrophages and cancer cells. nih.govnih.govresearchgate.net THP-1 cells can be differentiated into various macrophage populations, such as M0, M1, and M2, and co-cultured with cancer cells to evaluate cellular crosstalk. nih.govnih.govresearchgate.net While the provided search results primarily detail the use of THP-1 macrophages in the context of breast cancer cell interactions and not directly with this compound treatment, these cell systems represent a relevant model for future studies on how this compound or its analogs might influence macrophage function or their interaction with other cell types.

Ussing Chamber Electrophysiology

The Ussing chamber technique is a classical method used to measure ion transport across epithelial tissues or cultured cell monolayers. It allows for the measurement of transepithelial potential difference and short-circuit current, providing insights into the electrogenic transport of ions.

This compound is frequently used in Ussing chamber experiments as a specific blocker of epithelial sodium channels (ENaC) to isolate and study ENaC-mediated sodium transport. researchgate.netphysiology.orgfrontiersin.orgnih.gov For example, Ussing chamber studies on mucosal preparations of rat rectal colon showed that apical this compound inhibited a basal short-circuit current with an apparent half-inhibition constant (Ki) value of 0.20 μM. physiology.orgnih.gov In studies of human proximal tubule cells, Ussing chamber experiments demonstrated that this compound at a concentration of 10⁻⁴ M did not change the electrical properties, consistent with the known characteristics of proximal tubules. nih.gov The Ussing chamber is also used to study CFTR chloride current, where this compound is added to inhibit sodium transport at the apical side. researchgate.net

Ion-Selective Electrode Measurements

Ion-selective electrodes (ISEs) are electrochemical sensors used to measure the concentration or activity of specific ions in a solution. These electrodes generate a potential difference that is proportional to the logarithm of the ion activity.

ISEs have been developed and utilized for the determination of this compound itself, particularly in pharmaceutical samples. nahrainuniv.edu.iqnahrainuniv.edu.iqresearchgate.netscielo.br Studies have described the construction of this compound hydrochloride selective electrodes using polymeric membranes with different ion pairs and plasticizers. nahrainuniv.edu.iqresearchgate.net These electrodes exhibit characteristics such as slope, detection limit, and linear concentration range. nahrainuniv.edu.iqresearchgate.netscielo.br For instance, an this compound selective electrode based on salmon sperm ds-DNA as an ion pair, PVC matrix, and 2-nitrophenyl octylether (NPOE) as a plasticizer showed a Nernstian slope of 35.3 ± 1.0 mV decade⁻¹ and a detection limit of 1.5 × 10⁻⁶ mol L⁻¹. scielo.br

Here is a table summarizing some findings from this compound-selective electrode studies:

Note: Data for some parameters were not available in the provided snippets.

ISEs are also used in broader ion transport studies across epithelial cells, although detecting small changes in ion concentration can be technically challenging. mdpi.com

RNA-Seq, qRT-PCR, Immunoblotting, Immunofluorescence Assays

These molecular biology techniques are employed to investigate the effects of this compound on gene expression, protein levels, and cellular localization of specific molecules.

In studies of this compound's anti-myeloma activity, RNA-Seq, qRT-PCR, immunoblotting, and immunofluorescence assays were used to investigate its mechanisms of action. aacrjournals.orgresearchgate.net RNA-Seq experiments revealed that this compound significantly altered the level of transcript isoforms and alternative splicing events and deregulated the spliceosomal machinery. aacrjournals.orgresearchgate.net Immunofluorescence studies associated the disruption of the splicing machinery with the inhibition of myeloma cell viability after this compound exposure. aacrjournals.orgresearchgate.net Immunoblotting has also been used to examine the expression level of LAMP1, a marker of lysosomes, in cells treated with hexamethylene this compound (HA), an this compound derivative. d-nb.info Western blots have been used to characterize an this compound binding protein in adult alveolar type II pneumocytes using antibodies against epithelial Na+ channel proteins. nih.gov

Receptor Binding Profiling

Receptor binding profiling studies investigate the affinity and specificity of a compound for various receptors. This helps to understand the potential targets of a drug and its selectivity.

Studies have examined the receptor binding profiles of this compound and its analogues to investigate their interaction with different receptors, such as the adenosine-A₁ receptor. frontiersin.orgtandfonline.com These studies indicate that this compound and its analogues can affect radioligand binding to a variety of receptors, suggesting a widespread presence of an this compound binding site on receptors and other membrane proteins. tandfonline.com However, receptor binding profiles of this compound analogues have provided no evidence for a direct link between receptors and the Na+/H+ exchange, but rather indicate a common structure on receptor proteins. tandfonline.com An this compound binding protein with low affinity to benzamil and this compound has been characterized in adult alveolar type II pneumocytes. nih.gov

Molecular Docking and Computational Chemistry

Molecular docking and computational chemistry techniques are used to predict and analyze the binding interactions between this compound and its target molecules at an atomic level. These methods provide insights into binding sites, affinities, and the nature of interactions.

Molecular docking studies have been performed to investigate the interaction of this compound with various targets. This includes the interaction of this compound with the hASIC1a ion channel, using homology modeling of the pore region. worldscientific.com Docking results in this context agreed with the putative this compound binding site for alphaENaC channel chimeras. worldscientific.com Molecular docking has also been used to study the binding of this compound to human serum albumin (HSA), revealing that this compound primarily binds to Sudlow's site II, located in subdomain IIIA of HSA, with lesser affinity for Sudlow's site I. mdpi.com These studies utilize docking analysis to identify the location of binding and the residues involved in the interaction, such as van der Waals, hydrogen-bonding, and hydrophobic interactions. mdpi.com

Computational methods, including molecular docking and molecular dynamics simulations, have been employed to explore the potential inhibitory potency of this compound analogs against targets like the SARS-CoV-2 E viroporin. genominfo.org Docking simulations showed higher binding affinity scores for certain this compound analogs for SARS-CoV-2 E compared to SARS-CoV-1 E and indicated that these analogs engaged more amino acids in SARS-CoV-2 E. genominfo.org Molecular dynamics simulations further suggested that these ligands could alter the native structure and flexibility of the viral protein. genominfo.org Molecular docking has also been used to study the interaction of this compound with Acid-sensing ion channels (ASICs), showing that this compound interacts with specific amino acid residues. museu-goeldi.br Furthermore, molecular docking of modified this compound analogs has been used to study their binding properties to the NhaP-type cation-proton antiporter in Vibrio cholerae, revealing potential interactions with functionally important amino acid residues. cdnsciencepub.com

Here is a table summarizing some molecular docking findings for this compound:

Note: Specific interacting residues may vary depending on the protein structure and simulation conditions used in different studies.

Computational chemistry, including Density Functional Theory (DFT) calculations, is also used to provide insights into the electronic and structural properties of this compound and its interactions with targets like ASICs. museu-goeldi.br

Xenograft Mouse Models (e.g., Multiple Myeloma)

Xenograft mouse models, where human cancer cells are implanted into immunocompromised mice, have been used to investigate the anti-tumor activity of this compound. In the context of multiple myeloma (MM), this compound has shown potent anti-myeloma activity in xenograft mouse models. nih.gov Studies have observed this compound-induced apoptosis in these models. nih.gov Furthermore, this compound demonstrated a synergistic effect when combined with other agents like dexamethasone and melphalan in MM cell lines and xenograft models. nih.gov Research using MM xenograft models has also explored the effects of this compound analogs, such as hexamethylene this compound (HA), which has demonstrated significant anti-MM effects in vivo by inducing lysosome-mediated cell death. researcher.life

Genetically Modified Animal Models (e.g., βENaC-overexpressing mice)

Genetically modified animal models are valuable for studying the specific roles of ion channels and transporters targeted by this compound. While the search results did not provide specific details on studies using βENaC-overexpressing mice and this compound oup.com, this compound is known to inhibit the epithelial sodium channel (ENaC), which consists of alpha, beta, and gamma subunits. Studies using genetically modified models focusing on sodium channel activity or related pathways would be relevant for understanding this compound's effects on sodium transport and its physiological consequences.

Ischemic-Reperfused Rat Hearts

Ischemic-reperfused rat hearts are a model used to study myocardial injury and the effects of interventions. This compound, as an inhibitor of the Na+/H+ exchanger (NHE), has been investigated in this model due to the role of sodium and calcium overload in ischemia-reperfusion injury. Studies have shown that inhibition of the Na+/H+ exchanger by this compound or its derivatives, such as dimethyl this compound, can improve ischemia/reperfusion-induced ionic disturbance and contractile failure in perfused rat hearts. ahajournals.org This suggests a role for this compound in mitigating the effects of sodium accumulation during ischemia. ahajournals.org

Randomized Controlled Trials

Randomized controlled trials (RCTs) are considered the gold standard for evaluating interventions. This compound has been investigated in RCTs for several indications. For instance, a randomized, parallel, double-blinded, placebo-controlled trial (PREVER-prevention trial) investigated the effect of low-dose chlorthalidone in combination with low-dose this compound compared to placebo in patients with prehypertension. researchgate.net This trial demonstrated that the combination significantly reduced the incidence of hypertension and improved ECG-estimated left ventricular mass. researchgate.net

Another double-blind, placebo-controlled, crossover trial compared the effects of this compound and hydrochlorothiazide on glucose tolerance in patients with essential hypertension. researchgate.net This study found that this compound had a numerically lower blood pressure-lowering effect than hydrochlorothiazide, but the differences were not statistically significant over the 4-week treatment period. researchgate.net The study also observed a statistically significant rise in 30-minute insulin levels with this compound treatment compared to other treatments. researchgate.net

This compound is also being investigated in clinical trials for its potential neuroprotective effects, such as the this compound Clinical Trial In Optic Neuritis (ACTION) protocol, a randomized, double-blind, placebo-controlled trial assessing this compound's efficacy in acute optic neuritis. This trial aims to assess neuroprotection using measures like retinal nerve fiber layer thickness and various brain MRI outcomes.

A double-blind randomized clinical trial also compared the blood pressure-lowering efficacy of this compound versus enalapril as add-on therapy in patients with uncontrolled blood pressure receiving hydrochlorothiazide.

Data Table: PREVER-Prevention Trial Findings researchgate.net

Double-Blind, Placebo-Controlled Studies

Double-blind, placebo-controlled studies are considered a robust method for evaluating the effects of a treatment by minimizing bias. In such studies involving this compound, neither the participants nor the researchers are aware of whether the participant is receiving this compound or a placebo.

One double-blind, placebo-controlled, crossover trial investigated the effects of this compound compared to hydrochlorothiazide on glucose tolerance in patients with essential hypertension. This study involved two parts, with one part including 37 patients who received, in random order, 4 weeks of treatment with hydrochlorothiazide, nebivolol, a combination of hydrochlorothiazide and nebivolol, this compound, and placebo, each separated by a 4-week placebo washout period. ahajournals.orgnih.gov The primary outcome measured was the difference in glucose levels during a 2-hour oral glucose tolerance test between the start and end of the 4-week treatment periods when comparing hydrochlorothiazide and this compound. nih.gov The study found that for similar blood pressure reductions, there were opposite changes in glucose levels between this compound and hydrochlorothiazide, with this compound showing no impairment of glucose tolerance compared to placebo. ahajournals.orgnih.gov

Another randomized, double-blind, placebo-controlled trial, the PREVER-Prevention trial, evaluated the effectiveness of a low-dose combination of chlorthalidone and this compound for the prevention of hypertension in patients with prehypertension. ahajournals.org This study included participants aged 30 to 70 with prehypertension who were randomized to receive either the combination pill or placebo after 3 months of lifestyle intervention. ahajournals.org The incidence of hypertension was significantly lower in the group receiving the chlorthalidone/Amiloride combination compared to the placebo group. ahajournals.org

A double-blind randomized crossover trial in patients over 60 with hypertension compared this compound plus hydrochlorothiazide, atenolol, the combination of all three, and placebo. All active treatments reduced blood pressure, with the combination being significantly superior. doi.org

Data from a double-blind, placebo-controlled, crossover trial examining the influence of this compound on exercise tolerance in healthy adults showed that this compound significantly increased time to failure and total work performed during blood-flow-restricted plantar flexion compared to placebo, without significantly influencing mean blood pressure or heart rate responses. nih.gov

The this compound Clinical Trial In Optic Neuritis (ACTION) is a phase II randomized, double-blind, placebo-controlled trial investigating the neuroprotective efficacy of this compound in patients with acute optic neuritis. researchgate.netnih.govbmj.com The primary objective is to assess neuroprotection by measuring peripapillary retinal nerve fiber layer thickness at 6 months in the affected eye compared to the unaffected eye at baseline. researchgate.netnih.gov

Here is a table summarizing findings from some double-blind, placebo-controlled studies involving this compound:

Prospective Observational Studies

Prospective observational studies involve observing groups of participants over time to see if a particular factor is associated with an outcome. While not providing the same level of evidence as randomized controlled trials regarding causality, they can offer valuable insights into the effects of this compound in real-world settings or specific patient cohorts.

One prospective study in 46 kidney transplant recipients (KTRs) investigated the effect of dietary sodium intake reduction on systolic blood pressure, emphasizing the potential contribution of sodium retention to posttransplant hypertension. physiology.org Although this study primarily focused on dietary intervention, it highlights the context of sodium balance, which is relevant to this compound's mechanism of action as an epithelial sodium channel blocker. physiology.org

Another prospective randomized clinical trial conducted in South Korea investigated whether this compound was noninferior to spironolactone in reducing home-measured systolic blood pressure in patients with resistant hypertension. medscape.com This study, while randomized, is described as prospective and observational in its design regarding the collection of data over time in the defined patient group. It included 118 patients with resistant hypertension and found that this compound was noninferior to spironolactone in lowering home systolic blood pressure after 12 weeks. medscape.combktimes.net

Subgroup Analyses in Patient Populations

Subgroup analyses involve examining the effects of a treatment within specific subsets of the study population. This can help identify if this compound has differential effects based on patient characteristics, genetic factors, or disease subtypes.

A substudy of the PATHWAY-2 trial assessed the effect of this compound in a subgroup of 146 participants with resistant hypertension during an optional open-label run-out phase. bktimes.net This analysis showed that this compound reduced clinic systolic blood pressure. bktimes.net

In a study focusing on black hypertensive individuals with the T594M polymorphism, an open, controlled study design was used to assess the effect of this compound. ahajournals.org In this subgroup of 14 individuals, this compound alone effectively controlled blood pressure to a level similar to that achieved with their usual multiple-drug regimens. ahajournals.org When this compound was withdrawn, there was a significant increase in blood pressure, suggesting a particular effectiveness in this genetic subgroup. ahajournals.org This observation supports the concept that the T594M polymorphism may affect sodium channel activity and that this compound may help reverse this abnormality. ahajournals.org

In the prospective randomized clinical trial comparing this compound and spironolactone for resistant hypertension, a subgroup analysis indicated that spironolactone was more effective in patients with evidence of aldosterone excess, while this compound showed efficacy regardless of these factors. jwatch.org