Acarbose

Historical Trajectory of Discovery and Early Biochemical Characterization

Acarbose is a complex oligosaccharide that functions as an alpha-glucosidase inhibitor. Its origins trace back to microbial fermentation processes, specifically from cultures of the soil bacterium Actinoplanes utahensis drugs.comresearchgate.net. The discovery of this compound provided a novel approach to managing hyperglycemia by targeting carbohydrate digestion mdpi.com. Early biochemical characterization revealed its structure as a pseudotetrasaccharide, composed of an acarviosine moiety linked to maltose rcsb.orgwikipedia.org. Unlike typical saccharides, this compound contains a nitrogen linkage between the cyclitol (non-reducing end) and the deoxyhexose, which renders it resistant to cleavage by certain digestive enzymes researchgate.net. This unique structural feature contributes to its mechanism of action as an enzyme inhibitor researchgate.netrcsb.org. Further studies identified that this compound competitively and reversibly inhibits key enzymes involved in carbohydrate hydrolysis in the intestinal brush border drugs.comdrugbank.com.

The biosynthesis of this compound in Actinoplanes species has been a subject of extensive research. While the complete pathway was not initially understood, studies utilizing isotope tracers, gene inactivation, and biochemical experiments have progressively elucidated the enzymatic steps involved acs.orgnih.gov. It has been shown that the biosynthetic pathway starts from sedoheptulose 7-phosphate, an intermediate of the pentose phosphate pathway, and involves the incorporation of glutamate as the primary source of nitrogen frontiersin.org. Research has characterized various enzymes crucial for the formation and assembly of this compound's structural components, including kinases, phosphatases, nucleotidyltransferases, glycosyltransferases, and pseudoglycosyltransferases nih.gov.

This compound as a Prototypical Alpha-Glucosidase Inhibitor in Research

This compound is widely recognized as a prototypical compound within the class of alpha-glucosidase inhibitors and has been extensively studied in research wikipedia.orgnih.gov. Its well-characterized mechanism of competitive and reversible inhibition of intestinal alpha-glucosidases and pancreatic alpha-amylase has made it a benchmark for evaluating the activity of novel compounds with potential alpha-glucosidase inhibitory effects drugs.comdrugbank.comtandfonline.com.

Research studies investigating new potential alpha-glucosidase inhibitors often use this compound as a positive control for comparison of inhibitory potency (IC50 values) mdpi.comtandfonline.comresearchgate.net. For example, studies evaluating the alpha-glucosidase inhibitory activity of natural compounds or synthetic derivatives frequently report their efficacy relative to this compound tandfonline.comresearchgate.net. This comparative approach allows researchers to gauge the relative strength of inhibition and potential therapeutic relevance of novel agents tandfonline.comresearchgate.net.

Furthermore, this compound's interaction with alpha-glucosidase enzymes has been investigated at a molecular level. Structural studies, including those utilizing techniques like X-ray crystallography, have provided insights into how this compound binds to the active sites of these enzymes, revealing extensive hydrogen bonding interactions with key amino acid residues rcsb.org. These studies demonstrate that this compound acts by physically blocking the active sites, thereby preventing the binding and hydrolysis of natural oligosaccharide substrates rcsb.org.

Microbial Producers and Natural Isolation Sources

Acarbose is naturally produced by soil bacteria, notably strains belonging to the genus Actinoplanes. The most well-studied producer is Actinoplanes sp. SE50/110. researchgate.netdntb.gov.uaoregonstate.edufrontiersin.org Other Actinoplanes strains, such as Actinoplanes utahensis and Actinoplanes hulinensis, have also been identified as this compound producers. frontiersin.orgnih.gov Additionally, some Streptomyces species, such as Streptomyces glaucescens GLA.O and Streptomyces coelicoflavus ZG0656, produce acarviostatins, which are structurally related to this compound and share similar biosynthetic pathways. d-nb.infonih.govmdpi.com

This compound was initially extracted from the metabolites of Actinomycetes strains SE50, SE18, and SE82. mdpi.comgoogle.com These microorganisms serve as the natural isolation sources for this valuable compound.

Table 1: Key Microbial Producers of this compound and Related Compounds

Elucidation of Complete Biosynthetic Pathways and Intermediates

Significant progress has been made in elucidating the complete biosynthetic pathway of this compound in Actinoplanes sp. SE50/110. researchgate.netdntb.gov.uanih.govspringernature.com The pathway involves the synthesis and assembly of two main structural components: a C7-cyclitol moiety (valienol) and an amino-deoxyhexose unit, which are then linked to maltose. researchgate.netnih.gov

Identification and Characterization of Biosynthetic Gene Clusters (acb)

The genes responsible for this compound biosynthesis are clustered together in the genome of the producing organisms, forming the acb gene cluster. researchgate.netd-nb.infodntb.gov.uatandfonline.comresearchgate.netnih.gov In Actinoplanes sp. SE50/110, the acb gene cluster is approximately 32.2 kb in length and contains 22 genes. acs.org The sequencing and analysis of this gene cluster have been fundamental to understanding the biosynthetic pathway. tandfonline.comnih.gov While the acb cluster in Actinoplanes sp. SE50/110 lacks genes coding for transcriptional regulators, such regulators have been identified in the acarviostatin gene clusters of Streptomyces species. d-nb.info

Research has also focused on identifying essential genes within the acb cluster for this compound production. Studies using genome-scale metabolic models have indicated that specific acb cluster genes are essential for this compound synthesis, and the deletion of any of these genes can prevent production. frontiersin.org

Advanced Metabolic Engineering Strategies for Enhanced Research Production

Metabolic engineering approaches are crucial for overcoming bottlenecks in this compound production and enhancing yields in producing strains like Actinoplanes sp. SE50/110. acs.org These strategies involve rational design based on metabolic models and various genetic manipulation techniques.

Competitive and Reversible Inhibition Kinetics of Alpha-Glucosidases

Acarbose is characterized as a competitive and reversible inhibitor of intestinal alpha-glucosidases. drugbank.combjcardio.co.uknih.gov This means that this compound competes with the natural substrate for binding to the enzyme's active site. The inhibition is reversible because this compound can bind and unbind from the enzyme, and its effect can be overcome by increasing the concentration of the substrate. drugbank.combjcardio.co.uk

Enzyme Kinetic Parameters and Their Derivation (e.g., Km, Vmax, Ki)

Enzyme kinetics provide valuable insights into the mechanism of enzyme inhibition. Key parameters include the Michaelis constant (Km), maximum reaction velocity (Vmax), and the inhibitor dissociation constant (Ki). scispace.com Km represents the substrate concentration at which the reaction rate is half of Vmax, reflecting the enzyme's affinity for the substrate. Vmax is the maximum rate of the reaction when the enzyme is saturated with substrate. Ki is a measure of the inhibitor's potency, representing the dissociation constant of the enzyme-inhibitor complex. nih.gov A lower Ki value indicates a stronger binding affinity of the inhibitor for the enzyme.

Substrate Mimicry and Transition State Analog Characteristics

This compound's inhibitory potency stems from its structural resemblance to the natural substrates of alpha-glucosidases and alpha-amylase, particularly the transition state of the enzymatic reaction. wikipedia.orgrcsb.org this compound is described chemically as a pseudotetrasaccharide, mimicking a maltotetraose structure. wikipedia.org Its acarviosine moiety, which contains an amine linkage and a cyclohexene ring, is thought to mimic the transition state oxocarbenium ion intermediate that forms during the hydrolysis of glycosidic bonds by these enzymes. wikipedia.org

By mimicking the transition state, this compound binds to the enzyme's active site with high affinity, effectively "locking up" the enzyme and preventing the binding and hydrolysis of the natural substrate. wikipedia.org This transition state mimicry contributes significantly to this compound's potent inhibitory activity against alpha-glucosidases. rcsb.orgnih.govacs.org Structural studies, such as the crystal structure of alpha-glucosidase bound with this compound, have provided visual evidence of how this compound interacts with the enzyme's active site, occupying the substrate-binding pocket and mimicking the transition state conformation. rcsb.orgacs.org

Molecular Docking Studies of Acarbose with Alpha-Glucosidases

Molecular docking is a computational technique used to predict the preferred orientation (binding mode) of a ligand when bound to a protein target and to estimate the strength of the interaction (binding affinity). In studies investigating potential alpha-glucosidase inhibitors, this compound is frequently used as a reference compound or positive control due to its established inhibitory activity. mdpi.comnih.govmdpi.comherbmedpharmacol.comresearchgate.netwindows.netresearchgate.netresearchgate.netrsc.org Docking simulations predict how this compound fits into the active site of alpha-glucosidase enzymes.

Molecular Dynamics Simulations for Binding Stability and Conformational Changes

Comparative Molecular Modeling of this compound with Synthetic and Natural Inhibitors

Molecular modeling studies have frequently compared the binding interactions of this compound with those of other synthetic and natural alpha-glucosidase inhibitors to understand their relative potency and binding modes. These studies often involve docking simulations to predict the preferred orientation and binding affinity of inhibitors within the enzyme's active site, followed by molecular dynamics simulations to assess the stability of the resulting complexes.

Research has shown that this compound binds to the carbohydrate binding site of alpha-glucosidase, similar to its carbohydrate substrates. dergipark.org.tr The molecular structure of this compound, which resembles carbohydrates, allows it to bind to this site. dergipark.org.tr This binding is characterized by numerous hydrogen bonds and, in some cases, hydrophobic interactions with residues in the active site. researchgate.netnih.govjppres.com

Comparative studies using molecular docking have evaluated the binding potential of natural alpha-glucosidase inhibitors against this compound. Some natural compounds have demonstrated comparable or even better binding potential than this compound, forming various non-hydrogen bonds and exhibiting lower binding energy, suggesting a better binding affinity. dergipark.org.tr For instance, studies comparing this compound with flavonoids like quercetin have shown that quercetin can exhibit a higher binding affinity and interact with more catalytic residues than this compound in certain alpha-glucosidase enzymes. jppres.comjppres.com

Molecular dynamics simulations are used to assess the stability of the complexes formed between the enzyme and different inhibitors, including this compound. These simulations can reveal the dynamic behavior of the enzyme-inhibitor complex over time, providing insights into the stability of the binding interactions. dergipark.org.trnih.gov Studies have shown that some natural inhibitors can form complexes with similar stability to that of this compound. dergipark.org.tr

Crystallographic studies have provided high-resolution structures of alpha-glucosidase enzymes in complex with this compound, revealing the precise details of their interactions at the atomic level. For example, the crystal structure of Bacillus subtilis alpha-amylase in complex with this compound showed an this compound-derived hexasaccharide bound in the active site, occupying specific subsites. nih.govresearchgate.net These structures illustrate the extensive hydrogen bonding networks and other interactions that stabilize the this compound-enzyme complex. researchgate.netacs.org

Elucidation of Essential Structural Moieties for Alpha-Glucosidase Inhibition

SAR studies have aimed to identify the key structural features of acarbose responsible for its inhibitory activity against alpha-glucosidases. The pseudo-tetrasaccharide structure, particularly the acarviosine moiety, is considered essential for potent inhibition. researchgate.netresearchgate.net The presence of nitrogen within the acarviosine moiety, mimicking the transition state of substrate hydrolysis, contributes significantly to the high affinity of this compound for alpha-glucosidase enzymes. drugbank.comwikipedia.orgnih.govresearchgate.net

Based on research findings, the essential structural moieties and their proposed roles in alpha-glucosidase inhibition by this compound can be summarized as follows:

Impact of Chemical Modifications on Inhibitory Potency and Selectivity

In Silico Approaches to SAR Prediction and Optimization

Computational methods, such as molecular docking and quantitative structure-activity relationship (QSAR) analysis, play a significant role in predicting and understanding the SAR of this compound and its analogs. acgpubs.orgacs.orgdost.gov.phdergipark.org.trresearchgate.netmdpi.commdpi.combiorxiv.org These in silico approaches complement experimental studies by providing insights into the binding interactions between inhibitors and alpha-glucosidase enzymes at the molecular level. nih.govresearchgate.nettandfonline.comacgpubs.orgacs.orgdost.gov.ph

Molecular docking is widely used to predict the binding poses and affinities of this compound and its analogs within the active site of alpha-glucosidase. nih.govresearchgate.nettandfonline.comacgpubs.orgacs.orgdost.gov.phresearchgate.netmdpi.commdpi.com This technique helps visualize the interactions, such as hydrogen bonds and hydrophobic contacts, formed between the inhibitor and key amino acid residues in the binding pocket. researchgate.netresearchgate.netacs.orgdost.gov.phdergipark.org.trresearchgate.net By analyzing the predicted binding modes and energies, researchers can infer how structural modifications might affect the interaction with the enzyme and thus influence inhibitory potency. acgpubs.orgacs.orgdost.gov.phresearchgate.netmdpi.com

QSAR studies aim to build mathematical models that correlate structural or physicochemical properties of a series of compounds with their biological activity. acs.orgmdpi.com By developing QSAR models for this compound analogs, it is possible to identify molecular descriptors that are important for alpha-glucosidase inhibition and predict the activity of new, unsynthesized compounds. acs.org This can guide the design and synthesis of novel inhibitors with improved properties. acs.orgmdpi.com

Other computational techniques, such as molecular dynamics simulations and MM/GBSA calculations, are also employed to gain a more comprehensive understanding of the stability of the enzyme-inhibitor complex and the binding free energy. mdpi.combiorxiv.org These methods provide dynamic insights into the interactions over time and offer a more refined estimation of binding affinity. mdpi.combiorxiv.org

In silico studies have been used to compare the binding of this compound and its analogs to different alpha-glucosidase enzymes, including those from different species, helping to explain observed differences in inhibitory activity. scirp.org They can also be used to evaluate the selectivity of inhibitors towards alpha-glucosidase versus other enzymes like alpha-amylase. dergipark.org.tr

In Vitro Studies on Cellular Metabolism and Signaling Pathways

In vitro studies have begun to elucidate the direct effects of acarbose on cellular processes, particularly in the context of metabolic and vascular health. Research using vascular smooth muscle cells (VSMCs) has shown that this compound can inhibit migration and proliferation, key processes involved in the development of atherosclerosis. This effect appears to involve the modulation of specific signaling pathways. This compound has been found to upregulate microRNA-143 (miR-143) expression. MiR-143, in turn, negatively regulates Ras expression, impacting downstream signaling cascades such as the FAK/Src and PI3K/Akt pathways, which are critical for cell migration and proliferation nih.gov. This compound treatment reduced the phosphorylation of FAK and Src, as well as the expression of matrix metalloproteinases (MMPs), suggesting an attenuation of migration via the FAK/Src pathway. Additionally, this compound inhibited activated PI3K and Akt signals, along with Ras and PCNA expression and inactivated ERK signaling, indicating an impact on proliferation pathways nih.gov.

In the context of neuroprotection, in vitro studies using models of ischemia-reperfusion injury have demonstrated that this compound can confer significant neuroprotection. This protective effect is associated with the inhibition of the DAPK1-p53 interaction. This compound was found to prevent mitochondrial and lysosomal dysfunction and favorably modulate the expression of genes related to cell survival, inflammation, and regeneration. Furthermore, this compound treatment increased cell proliferation and differentiation in these in vitro models researchgate.netnih.gov.

Animal Model Research on Metabolic Homeostasis Beyond Glucose Absorption

Animal studies have been instrumental in revealing the broader metabolic effects of this compound, extending beyond its well-known inhibition of glucose absorption in the small intestine.

Immunomodulatory Effects in Preclinical Models

Preclinical evidence suggests that this compound possesses immunomodulatory properties. In addition to its role in enhancing anti-tumor immunity through modulation of the gut microbiota and CD8 T cell responses mdpi.comresearchgate.netresearchgate.net, this compound has been investigated for its effects on inflammatory and autoimmune conditions.

Studies in a mouse model of collagen-induced arthritis (CIA), a model for rheumatoid arthritis, have shown that this compound can attenuate the incidence and severity of arthritis researchgate.netfrontiersin.org. This was associated with a reduction in pro-inflammatory cytokines, including TNF-α, IL-6, and IL-17, in paw tissues researchgate.net. This compound treatment also decreased the production of anti-collagen type II antibodies and reduced the levels of IL-17 and IFN-γ produced by collagen-reactive lymph node cells researchgate.net. The beneficial effects in this arthritis model were linked to the restoration of gut microbiota homeostasis and changes in the balance of T helper 17 (Th17) and regulatory T (Treg) cells in the gut frontiersin.org.

Furthermore, a combination therapy of this compound and cyclosporine A ameliorated imiquimod-induced psoriasis-like dermatitis in mice. This combination significantly reduced the percentages of IL-17- and IL-22-producing CD4+ T cells (Th17 and Th22 cells) and increased the percentage of Treg cells, suggesting an anti-inflammatory mechanism involving the modulation of T cell subsets nih.gov.

These preclinical findings highlight the diverse immunomodulatory effects of this compound, potentially mediated through its interaction with the gut microbiome and direct effects on immune cells and cytokine production.

Role of Intestinal Bacteria in Acarbose Degradation

Intestinal bacteria play a crucial role in the degradation of this compound. fda.govnih.govwikipedia.org this compound increases the amount of undigested starch that reaches the colon, thereby enriching carbohydrate-degrading bacteria and altering the gut microbiota composition and its fermentation products. aginganddisease.orggethealthspan.com This can lead to an increase in the abundance of bacteria capable of producing short-chain fatty acids (SCFAs). aginganddisease.orggethealthspan.com

Specific gut bacteria, such as Klebsiella grimontii TD1, have been identified as this compound-degrading organisms. researchgate.net The presence of such bacteria is associated with variability in the clinical efficacy of this compound. researchgate.netresearchgate.net Studies have shown that K. grimontii TD1 can effectively metabolize this compound. researchgate.net

This compound also influences the composition of the gut microbiota, increasing the relative abundance of saccharolytic bacteria like Lactobacillus and Bifidobacterium, while potentially depleting other species such as Bacteroides, Alistipes, and Clostridium. gutmicrobiotaforhealth.com The effect of this compound on intestinal fermentation products, such as SCFAs, can influence host physiology. aginganddisease.org SCFAs, primarily produced by the fermentation of starch by intestinal bacteria, are considered beneficial to health. aginganddisease.org

Research using model starch-degrading Bacteroides species, Bacteroides ovatus (Bo) and Bacteroides thetaiotaomicron (Bt), has shown differential responses to this compound. asm.orgnih.govbiorxiv.org While this compound interferes with the ability of some bacteria to grow on starch, the specific mechanisms and species-level variations are complex. asm.orgnih.govbiorxiv.org

Enzymatic Biotransformation Pathways

This compound is metabolized within the gastrointestinal tract through enzymatic hydrolysis, primarily by intestinal bacteria and to a lesser extent by digestive enzymes. fda.govnih.govdrugbank.com Human enzymes do contribute to this compound transformation; for instance, pancreatic alpha-amylase can perform a rearrangement reaction on the molecule. wikipedia.org However, bacterial alpha-amylases from the gut microbiome are also capable of degrading this compound. wikipedia.org

A key enzymatic pathway involves glycoside hydrolases produced by gut microbiota. wikipedia.org Specifically, an this compound-degrading glucosidase, termed Apg, has been identified in Klebsiella grimontii TD1. researchgate.net This enzyme hydrolyzes this compound into smaller molecules. researchgate.net

While this compound is known to inhibit host enzymes like alpha-glucosidase and pancreatic alpha-amylase, gut bacteria possess their own enzymatic systems, such as the starch utilization system (Sus) in Bacteroides, which can interact with this compound. asm.orgnih.govbiorxiv.org Studies suggest that this compound can target periplasmic GH97 enzymes within Bacteroides and affect starch processing at multiple points, including competing for transport and binding to transcriptional regulators. asm.orgnih.govbiorxiv.org

The biosynthesis of this compound itself is a complex process carried out by microorganisms like Actinoplanes sp. SE50/110. nih.govnih.govresearchgate.netfrontiersin.orgresearchgate.net This involves a series of enzymatic reactions, including those catalyzed by cyclitol modifying enzymes, glycosyltransferases (e.g., AcbI), and pseudoglycosyltransferases (e.g., AcbS). nih.govresearchgate.netfrontiersin.org Understanding the microbial biosynthetic pathways provides insight into the structural components and potential degradation targets of this compound.

Characterization of Metabolites and Their Biological Activity

This compound is extensively metabolized in the gastrointestinal tract, yielding at least 13 identified metabolites that have been separated chromatographically from urine specimens. fda.govdrugbank.com Approximately one-third of these metabolites are absorbed into the systemic circulation and subsequently excreted renally. drugbank.com

The major metabolites identified are 4-methylpyrogallol derivatives, which exist as sulfate, methyl, and glucuronide conjugates. fda.govdrugbank.com

Research has characterized the degradation products of this compound mediated by bacterial enzymes like Apg from K. grimontii TD1. researchgate.netresearchgate.net Apg degrades this compound into acarviosine-glucose and acarviosine. researchgate.netresearchgate.net

Studies have investigated the biological activity of these metabolites. One metabolite, formed by the cleavage of a glucose molecule from this compound, has been shown to retain some alpha-glucosidase inhibitory activity. fda.govdrugbank.comnih.gov However, the degradation products acarviosine-glucose and acarviosine exhibit significantly reduced or almost complete loss of inhibitory activity on alpha-amylase compared to the parent compound this compound. researchgate.netresearchgate.net This loss of inhibitory function in metabolites contributes to the mechanism by which bacterial degradation can potentially reduce the efficacy of this compound. researchgate.net

While the parent compound this compound is the primary active entity responsible for inhibiting intestinal alpha-glucosidases and pancreatic alpha-amylase, the formation and absorption of metabolites, such as the 4-methylpyrogallol derivatives, indicate systemic exposure to these transformation products. fda.govdrugbank.com

Here is a table summarizing the key metabolites and their reported activity:

High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC)

High-Performance Liquid Chromatography (HPLC) and its faster, higher-resolution counterpart, Ultra-Performance Liquid Chromatography (UPLC), are indispensable tools in acarbose research. These techniques are widely used for the separation, identification, and quantification of this compound in various matrices, including bulk drug material, pharmaceutical formulations, and biological samples. HPLC methods for this compound often utilize reversed-phase chromatography with specific columns designed for carbohydrate analysis, such as amino silane bonded silica gel or carbohydrate columns. nih.govresearchgate.netgoogle.com Detection is commonly achieved using UV detectors, typically at wavelengths around 210 nm, although this compound itself has a weak chromophore, which can pose challenges for UV detection of certain impurities. google.comresearchgate.netthermofisher.com Alternative detection methods like Charged Aerosol Detection (CAD) or Evaporative Light Scattering Detection (ELSD) are also employed, offering more universal detection capabilities for compounds lacking strong UV absorbance. thermofisher.comnih.gov

UPLC offers significant advantages over conventional HPLC, including shorter analysis times, increased sensitivity, and improved resolution, making it particularly useful for analyzing complex samples and detecting low-level impurities. scispace.commdpi.com

Spectroscopic Techniques (e.g., Fourier Transform Infrared Spectroscopy (FTIR), Nuclear Magnetic Resonance (NMR), Fluorescence Spectroscopy)

Spectroscopic techniques provide valuable information about the structure, functional groups, and interactions of this compound.

Fourier Transform Infrared Spectroscopy (FTIR) can be used for the quantitative determination of this compound in tablets. alliedacademies.org An ATR-FTIR method has been developed that allows for the direct analysis of finely ground tablets without extensive sample preparation, offering a fast and non-destructive alternative to chromatographic methods for quality control. alliedacademies.org

Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful technique for the structural elucidation of this compound and its impurities. nih.govcapes.gov.brnih.gov Both 1D and 2D NMR experiments, such as 1H and 13C NMR, provide detailed information about the arrangement of atoms and functional groups within the molecule. LC-NMR, which couples liquid chromatography with NMR, allows for the online separation and structural identification of components in complex mixtures, proving useful for impurity profiling of this compound. nih.govcapes.gov.br

Fluorescence spectroscopy can be applied in research involving this compound, particularly in studies investigating its interaction with enzymes or other biological molecules. While this compound itself may not be strongly fluorescent, its interaction with fluorescent probes or enzymes can be monitored using this technique. rsc.orgspectroscopyonline.com For example, fluorescence spectroscopy has been used to study how this compound affects the functional abilities of enzymes like α-amylase. spectroscopyonline.com

Capillary Electrophoresis (CE)

Capillary Electrophoresis (CE) is another separation technique utilized in this compound analysis. CE offers high separation efficiency and can be applied to the analysis of this compound and its related substances. researchgate.netnih.govresearchgate.netingentaconnect.com Methods based on capillary zone electrophoresis (CZE) have been developed for the determination of this compound, sometimes involving derivatization to facilitate detection, such as with 7-aminonaphthalene-1,3-disulfonic acid (ANDS). researchgate.netresearchgate.netingentaconnect.comnih.gov Alternatively, detection can be achieved by monitoring the unsaturated cyclitol moiety of this compound at low wavelengths (below 200 nm). researchgate.netresearchgate.netingentaconnect.com CE methods can be validated for parameters like precision, accuracy, and linearity, and have been successfully applied to the analysis of this compound in pharmaceutical formulations. nih.govingentaconnect.com Affinity capillary electrophoresis (ACE) has also been employed to study the interaction of this compound with proteins like human serum albumin (HSA).

Enzyme-Based Biosensors for this compound Detection

Enzyme-based biosensors offer a different approach for the detection and study of this compound, leveraging its inhibitory effect on enzymes like alpha-glucosidase. researchgate.netrsc.orgmdpi.comingentaconnect.com These biosensors typically incorporate an enzyme, such as alpha-glucosidase, immobilized on a transducer. mdpi.comnih.gov The principle of detection often relies on monitoring the enzyme's activity in the presence of a substrate and observing the reduction in activity caused by this compound's inhibitory action. rsc.orgnih.gov Electrochemical biosensors have been developed for this purpose, where the enzymatic reaction produces a detectable electrochemical signal that changes upon inhibition by this compound. mdpi.comnih.gov These biosensors can be used for screening potential alpha-glucosidase inhibitors, including this compound and compounds from natural sources. rsc.orgmdpi.com

Mass Spectrometry (MS) for Structural Elucidation and Impurity Profiling

Mass Spectrometry (MS) is a powerful technique for determining the mass-to-charge ratio of molecules, providing crucial information for structural elucidation and impurity profiling of this compound. xml-journal.netnih.govcapes.gov.brnih.gov When coupled with separation techniques like HPLC or UPLC (LC-MS or UPLC-MS), MS allows for the analysis of complex mixtures, enabling the identification of this compound and its related impurities based on their mass fragmentation patterns. xml-journal.netnih.govcapes.gov.br

LC-MS and LC-MS/MS (tandem mass spectrometry) are particularly valuable for identifying and characterizing impurities in this compound bulk drug material and formulations. xml-journal.netnih.govcapes.gov.br This integrated approach allows for the online separation of impurities by chromatography followed by their structural investigation using MS. nih.govcapes.gov.br By analyzing the fragmentation patterns, researchers can deduce the structures of unknown impurities, including structural analogs that may differ from this compound in their sugar subunits. nih.govcapes.gov.br MS can also be used for impurity profiling, providing a comprehensive picture of the related compounds present in a sample. nih.gov

The combination of LC-NMR and LC-MS has proven particularly effective for the comprehensive impurity profiling and structural elucidation of components in this compound samples. nih.govcapes.gov.br

Interactions with Natural Products and Phytochemicals

Studies have investigated the combined effects of acarbose with various natural products and phytochemicals on the inhibition of carbohydrate-hydrolyzing enzymes like α-glucosidase and α-amylase. These combinations can lead to synergistic or additive inhibitory effects, offering potential for improved glycemic control.

Specific phytochemicals, including flavonoids like baicalein, apigenin, quercetin, luteolin, and (+)-catechin, have been studied for their interactions with this compound against α-glucosidase and α-amylase. For instance, combinations of baicalein, quercetin, or luteolin with this compound have shown synergistic inhibition of α-glucosidase. acs.org Conversely, the combination of (+)-catechin with this compound exhibited antagonistic inhibition of α-glucosidase. acs.org Apigenin and baicalein have also shown interactions with this compound against α-amylase, with combinations showing additive inhibition at lower concentrations and antagonistic inhibition at higher concentrations. acs.org

Cyanidin-3-galactoside, a natural anthocyanin, has also demonstrated synergistic inhibitory effects on intestinal α-glucosidase (maltase and sucrase) when combined with a low dose of this compound. tandfonline.com The percentage inhibition of the mixtures was significantly increased compared to this compound alone. tandfonline.com

Other natural compounds, such as betulinic acid (a pentacyclic triterpene), have shown a synergistically inhibitory effect when combined with this compound against α-glucosidase. semanticscholar.org Similarly, studies on Thai folk anti-diabetes remedies have indicated that certain decoction extracts can exhibit potent synergistic inhibitory effects with this compound on α-glucosidase. tjnpr.org

Molecular docking studies have been employed to understand the interactions of phytochemicals with enzymes targeted by this compound. For example, naringin and rutin from Capparis spinosa have shown high affinities to both α-amylase and α-glucosidase, comparable to that of this compound, suggesting they could work synergistically. rjpbcs.com

Combinatorial Effects with Other Enzyme Inhibitors (Preclinical/Mechanistic)

Beyond combinations with natural products, research has also explored the combinatorial effects of this compound with other synthetic enzyme inhibitors at a preclinical and mechanistic level. These studies aim to understand if combining this compound with other inhibitors targeting the same or different enzymes involved in carbohydrate metabolism can lead to improved or altered inhibitory profiles.

This compound is known to competitively and reversibly inhibit both pancreatic α-amylase and membrane-bound intestinal α-glucosidases. drugbank.com Its inhibitory potency against α-glucosidases follows the order of glucoamylase > sucrase > maltase > isomaltase. drugbank.com Combining this compound with other inhibitors that target these same enzymes but potentially through different mechanisms or at different binding sites could lead to enhanced or synergistic inhibition.

Preclinical studies investigating the combination of this compound with other enzyme inhibitors focus on the mechanistic aspects of their interaction and the resulting impact on enzyme activity. While specific examples of combinatorial effects with synthetic enzyme inhibitors beyond those naturally derived are less prominent in the provided search results, the principle of combining inhibitors with different binding sites or mechanisms of action, as seen with natural products, is relevant. For instance, the observation that this compound (competitive inhibitor) and betulinic acid (non-competitive inhibitor) bind to different sites and exhibit synergistic effects provides a mechanistic basis for exploring combinations with other non-competitive or allosteric inhibitors. semanticscholar.org

The concept of combining inhibitors to achieve better therapeutic outcomes is also highlighted in the context of overcoming limitations of single-drug therapy, such as potential drug tolerance or insufficient efficacy. jyoungpharm.org Exploring combinations of this compound with other enzyme inhibitors at a preclinical level allows for the investigation of potential synergistic interactions, altered kinetic profiles, and the molecular basis of these combined effects before moving to clinical studies.

Exploration of Novel Biological Targets and Non-Canonical Mechanisms

Beyond its classical inhibition of alpha-glucosidases and alpha-amylase, research is delving into previously unrecognized biological targets and non-canonical mechanisms of acarbose. Studies suggest that this compound may exert effects through pathways independent of its direct enzymatic inhibition in the small intestine. One area of investigation involves the potential of this compound to influence the gut microbiome, which in turn can impact metabolic processes and potentially contribute to its observed benefits, including weight management and effects on glucagon-like peptide-1 (GLP-1) levels. nih.govclinicaltrials.gov The interaction with gut bacteria and the potential inactivation of this compound by microbiome-derived enzymes like this compound kinases (Maks) represent a significant area of emerging research, highlighting a complex interplay between the drug and the host microbiome. princeton.edunih.gov Furthermore, research is exploring the impact of this compound on intracellular signaling pathways, such as the MAPK pathways, which are involved in cellular responses to insulin/IGF1 signals and inflammatory processes. nih.gov The potential for this compound to act as a "caloric restriction mimetic" by mimicking some of the metabolic changes associated with caloric restriction, independent of substantial net caloric reduction, is also being investigated as a potential non-canonical mechanism contributing to its effects, including potential links to longevity. clinicaltrials.govresearchgate.net Additionally, this compound is being explored for its potential to target alpha-glucosidase in Candida albicans, suggesting a possible role in mitigating candidiasis by affecting fungal cell wall integrity and virulence factors like biofilm formation and morphological switching. nih.gov

Development of this compound-Derived Probes for Biochemical Studies

The structural features of this compound, particularly its pseudo-oligosaccharide nature and inhibitory activity, make it a valuable scaffold for developing biochemical probes. These probes can be utilized to investigate the activity and structure of glycosidases and amylases, as well as to explore their interactions with inhibitors. Research involves designing and synthesizing this compound analogs with modifications that allow for labeling or immobilization, facilitating studies on enzyme kinetics, binding sites, and mechanisms of action. The development of activity-based protein profiling (ABPP) probes based on this compound-like structures is an example of this direction, enabling the selective labeling and study of retaining amylases from various biological sources. researchgate.net Such probes can aid in the discovery of new enzyme inhibitors and in understanding the functional roles of glycosidases in different biological systems.

Integration of Multi-Omics Data in this compound Research

The advent of multi-omics technologies (genomics, transcriptomics, proteomics, metabolomics, and microbiomics) is providing unprecedented opportunities to gain a holistic understanding of this compound's effects at a systems level. Integrating data from these diverse platforms allows researchers to explore how this compound influences gene expression, protein profiles, metabolic pathways, and the composition and function of the gut microbiome. nih.govnih.govrsc.org This integrated approach can help identify novel biomarkers of response to this compound, elucidate the complex interplay between the drug, the host, and the microbiome, and uncover previously unknown mechanisms of action. For instance, multi-omics data integration is being used to study the impact of this compound on the kidneys in Rattus norvegicus, revealing shifts in gene expression cascades and pathway enrichment. researchgate.net Challenges in multi-omics data integration, such as standardization and data analysis, are being addressed with advanced computational methods. nih.govrsc.org

Advanced Computational Design of this compound Analogs and Mimetics

Computational approaches are playing an increasingly vital role in the design and discovery of novel this compound analogs and mimetics with potentially improved efficacy, specificity, or reduced side effects. Techniques such as molecular docking, molecular dynamics simulations, and quantitative structure-activity relationship (QSAR) modeling are employed to predict the binding affinity and inhibitory potential of designed compounds against target enzymes like alpha-glucosidase and alpha-amylase. herbmedpharmacol.comacs.orgmdpi.comnih.gov Advanced computational design, including the use of machine learning and generative models, is being explored to generate novel molecular structures that mimic the inhibitory properties of this compound or target alternative pathways. mathiasbernhard.chtuwien.ac.at These computational methods can significantly accelerate the lead discovery process and guide the synthesis of promising new compounds. Studies have utilized computational docking to evaluate the binding of potential inhibitors from natural sources, using this compound as a reference standard. mdpi.comresearchgate.net

Q. What are the primary mechanisms of action of Acarbose in glycemic control, and how are these mechanisms experimentally validated?

this compound inhibits intestinal α-glucosidase enzymes, delaying carbohydrate digestion and reducing postprandial hyperglycemia. Methodologically, this is validated via in vitro enzyme assays (e.g., PNPG substrate hydrolysis) and kinetic analyses (Lineweaver-Burk plots) to determine inhibition type (competitive/mixed) . Clinical trials corroborate this by measuring postprandial glucose reduction (e.g., 1.63–3.62 mmol/L decrease with 50–300 mg doses) .

Q. What experimental designs are standard for evaluating this compound efficacy in clinical trials?

Randomized, double-blind, placebo-controlled trials (RCTs) with impaired glucose tolerance (IGT) or type 2 diabetes (T2D) cohorts are standard. Key endpoints include:

• Primary : Glycated hemoglobin (HbA1c) reduction, diabetes incidence .
• Secondary : Postprandial glucose, cardiovascular events, hypertension . The STOP-NIDDM trial (n=1,368, 3.3-year follow-up) exemplifies this design, using intent-to-treat analysis and hazard ratios for risk reduction .

Q. How is this compound dosage optimized in pharmacological studies, and what are common dose-response relationships?

Dosage optimization involves titration (e.g., 50–300 mg t.i.d.) with incremental HbA1c reductions. A meta-analysis showed dose-dependent postprandial glucose lowering (1.63 mmol/L at 50 mg vs. 3.62 mmol/L at 300 mg) . Trials often use 100 mg t.i.d. for balancing efficacy and gastrointestinal tolerability .

Q. What quality control methods ensure this compound purity in pharmaceutical research?

Pharmacopeial standards include:

• Purity : HPLC with UV detection, comparing peak areas of related substances (e.g., ≤0.5% impurities) .
• Residual solvents : Heavy metal testing via atomic absorption spectroscopy (≤10 ppm lead) .

Advanced Research Questions

Q. How do molecular docking studies resolve contradictions in this compound’s inhibition type (competitive vs. mixed)?

Discrepancies arise from methodological differences: in silico docking often predicts competitive inhibition by binding active sites, while in vitro kinetics (e.g., Hanes-Wolff plots) may suggest mixed inhibition due to allosteric effects. Multi-method validation (e.g., Eisenthal-Cornish-Bowden plots) is critical .

Q. What molecular pathways beyond α-glucosidase inhibition contribute to this compound’s metabolic effects?

this compound upregulates miR-10a-5p and miR-664 in the ileum, suppressing proinflammatory cytokines (e.g., TNF-α) via MAPK pathways. This is validated through miRNA sequencing and Western blotting in diabetic rat models .

Q. How do researchers address conflicting data on this compound’s cardiovascular benefits across meta-analyses and RCTs?

The MERIA meta-analysis (7 RCTs) reported cardiovascular risk reduction (HR 0.65), while individual trials like STOP-NIDDM showed 49% risk reduction. Heterogeneity arises from population differences (e.g., baseline HbA1c, IGT vs. T2D) and endpoint definitions (e.g., composite vs. individual events) . Sensitivity analyses and subgroup stratification (e.g., HbA1c ≥7%) mitigate these issues .

Q. What advanced analytical techniques quantify this compound’s pharmacokinetics and pharmacodynamics?

• Pharmacokinetics : LC-MS/MS for plasma concentration profiling, focusing on low bioavailability (<2%) due to minimal systemic absorption .
• Pharmacodynamics : Continuous glucose monitoring (CGM) to track postprandial fluctuations, combined with mechanistic modeling (e.g., HOMA-IR for insulin resistance) .

Q. How is this compound integrated into combination therapy regimens, and what methodological considerations apply?

In metformin-insufficient T2D patients, add-on this compound (100 mg b.i.d.) reduced HbA1c by 1.02% (P=0.0001) in a 24-week RCT. Key considerations include:

• Titration : Gradual dose escalation to minimize GI adverse events.
• Endpoint selection : Composite metrics (e.g., HbA1c + fasting glucose) for holistic efficacy assessment .

Q. What surrogate markers validate this compound’s long-term cardioprotective effects in preclinical studies?

Carotid intima-media thickness (CIMT) is a key surrogate; in STOP-NIDDM, this compound reduced CIMT progression by 50% (0.02 mm vs. 0.05 mm in placebo, P=0.027) via postprandial glucose modulation .

Methodological Tables

Q. Table 1: Dose-Dependent Efficacy of this compound in Postprandial Glucose Reduction

Q. Table 2: Key Outcomes from the STOP-NIDDM Trial