Glibenclamide Research Articles

Glibenclamide-encapsulated Liposomes alleviate LPS-induced Inflammatory Cascade through NLRP3 inhibition in macrophages

Abstract Inflammation is a vital immune response for survival during infection and tissue damage. It is critical in maintaining normal tissue homeostasis by orchestrating appropriate inflammatory mediators. Macrophages, integral to innate immunity, respond to lipopolysaccharide (LPS) present in gram-negative bacteria by releasing inflammatory cytokines. Utilizing nanotechnology for drug delivery have been proven with enhanced therapeutic approaches by targeting the suppression of inflammatory mediators and cytokines. Recent studies have provided insights into the role of inflammasomes in intracellular processes linked to inflammation. Glibenclamide (GLB), a sulfonylurea used in type 2 diabetes treatment, has emerged as a potent inhibitor of the NLRP3 inflammasome, showing promise in alleviating inflammation-associated injuries. To overcome the limitations of GLB, such as low aqueous solubility and high permeability, in this study, methyl-PEG-DSPE lipids were used to develop GLB-loaded nanoliposomes. The size of blank liposome was measured to be 120 nm. Anionic GLB-loaded liposomes, sized 146 nm with spherical morphology, effectively suppressed the expression of NLRP3 mediators (caspase-1, ASC, IL-1B, and IL-18) and various reactive oxygen species mediators compared to free GLB, reducing LPS-induced inflammatory responses in RAW264.7 macrophages. This suggests the potential of GLB-loaded liposomes as a therapeutic agent for inflammation-related disorders, warranting further in-vivo investigation.

Optimization of Glibenclamide Loaded Thermoresponsive SNEDDS Using Design of Experiment Approach: Paving the Way to Enhance Pharmaceutical Applicability.

Thermoresponsive self-nanoemulsifying drug delivery systems (T-SNEDDS) offer a promising solution to the limitations of conventional SNEDDS formulations. Liquid SNEDDS are expected to enhance drug solubility; however, they are susceptible to leakage during storage. Even though solid SNEDDS offers a solution to this storage instability, they introduce new challenges, namely increased total dosage and potential for drug trapping within the formulation. The invented T-SNEDDS was used to overcome these limitations and improve the dissolution of glibenclamide (GBC). Solubility and transmittance studies were performed to select a suitable oil and surfactant. Design of Experiments (DoE) software was used to study the impact of propylene glycol and Poloxamer 188 concentrations on measured responses (liquefying temperature, liquefying time, and GBC solubility). The optimized formulation was subjected to an in vitro dissolution study. The optimized T-SNEDDS consisted of Kolliphor EL and Imwitor 308 as surfactants and oil. The optimized propylene glycol and Poloxamer 188 concentrations were 13.7 and 7.9% w/w, respectively. It exhibited a liquefying temperature of 35.0 °C, a liquefying time of 119 s, and a GBC solubility of 5.51 mg/g. In vitro dissolution study showed that optimized T-SNEDDS exhibited 98.8% dissolution efficiency compared with 2.5% for raw drugs. This study presents a promising approach to enhance pharmaceutical applicability by resolving the limitations of traditional SNEDDS.

Self-Nanoemulsifying Drug Delivery System Combined with a Polymeric Amorphous System of Glibenclamide for Enhanced Drug Dissolution and Stability.

Self-nanoemulsifying drug delivery systems (SNEDDS) have been widely applied to improve the dissolution and bioavailability of hydrophobic medications like glibenclamide (GB). However, the acid liability of GB limits its loading in SNEDDS formulation owing to the expected drug degradation. The present study investigated the ability of a polymeric amorphous system (PAS) to amorphize raw GB and facilitate its integration within dispersed SNEDDS. Liquid-SNEDDS (L-SNEDDS), solid-SNEDDS (S-SNEDDS), and combined systems (SNEDDS + PAS) were prepared for this purpose. The physicochemical properties of the prepared formulations were examined using a zeta-sizer, SEM, DSC, PXRD, and dissolution apparatus. In addition, GB integrity within formulations following incubation in a stability chamber was also investigated. The prepared formulations were able to be dispersed within the nanosize range. SEM, DSC, and PXRD showed that freeze-drying (FD) was superior to the microwave (MW) method in GB amorphization. Even though L-SNEDDS and S-SNEDDS were able to increase the dissolution efficiency (DE) of GB, drug degradation was observed. However, PAS prepared using FD was able to increase the DE of GB from 2.5% to 84.2% and protect the drug from chemical degradation. The present study revealed that a combined system (SNEDDS + PAS) is a promising approach to enhance the stability of acid-labile drugs and facilitate the integration of amorphous drugs within a dispersed SNEDDS formulation.

Identification and in vitro genotoxicity assessment of forced degradation products of glimepiride and glyburide using HEK cell-based COMET assay.

This study focuses on characterizing the forced degradation products of antidiabetic drugs glimepiride (GMD) and glyburide (GBD), with previously unexplored genotoxicity. Drugs underwent stress induced by acid, base, and hydrogen peroxide. For GMD, impurities were profiled and isolated using Hypersil Gold C8 (250 × 10 mm, 5μ) through semi-preparative HPLC with a fraction collector. For GBD, impurity profiling was performed using semi-preparative HPLC (Hypersil GOLD C18, 250 × 10 mm, 5μ), and reverse-phase flash chromatography (FP ECOFLEX C18 4g column) for isolation. Although five GMD and three GBD impurities were detected, only three GMD and two GBD impurities were separated and assessed for purity using analytical RP-HPLC with the purity percentages ranging from 96.6% to 99.9%. LC-Orbitrap MS was used to identify these three GMD impurities (m/z: 408.122, 338.340, 381.160) and two GBD impurities (m/z: 369.065, 325.283). ProTox-II in silico predictions classified all impurities as class 4 and 5, with no positive genotoxicity indications. In vitro comet assays, using HEK cells, indicated that for GMD, impurity 2 and impurity 5 were less genotoxic, whereas impurity 4 exhibited genotoxicity. For GBD, both impurities 1 and 3 were found to be genotoxic, with impurity 3 showing a higher level of genotoxicity than impurity 1.

Mechanisms of dissolution and crystallization of amorphous glibenclamide

Amorphous solid dispersions enhance the dissolution and oral bioavailability of poorly water-soluble drugs. However, the link between polymer properties and formulation performance has not been fully clarified yet. We studied the effect of hydroxypropyl cellulose (HPC) polymers molecular weight (Mw) on the storage stability, dissolution kinetics and supersaturation stability of spray-dried amorphous glibenclamide (GLB) formulations. The solid-state stability of amorphous GLB during storage was significantly enhanced by both the 40 kDa (HPC-SSL) and 84 kDa (HPC-L) polymers, regardless of Mw differences. In contrast, HPC-SSL maintained significantly higher aqueous drug concentrations during dissolution, compared to HPC-L (its higher Mw analogue). Dedicated dissolution experiments, in situ optical microscopy and solid-state characterization revealed that aqueous drug concentrations were determined by the interplay between crystallization inhibition, drug ionization, wetting and solubilization effects: (1) HPC prevents surface nucleation, hence inhibiting crystallization, (2) intestinal colloids (bile salts and phospholipids) increase supersaturated drug concentrations via wetting and solubilization effects and (3) pH and drug ionization severely impact the degree of supersaturation. The better performance of the lower Mw HPC-SSL was due to its superior inhibition of surface crystallization. These insights into the molecular mechanisms of dissolution and crystallization of amorphous solids provide foundation for rational formulation development.

The effectiveness of edible bird's nest in lowering VEGF, CD31, and PDGFR-β levels in diabetic retinopathy in rats with type 1 diabetes.

Diabetic eye disease, known as diabetic retinopathy (DR), is one of the problems that can arise from having high blood sugar for an extended period. This study aimed to investigate the effect of the edible bird's nest (EBN) on retinal angiogenesis in diabetic rats. The 50 rats were separated into five different groups, each containing 10 rats: control, diabetes (DM), bird's nest-fed diabetes (75 mg/kg Body weight; BW), (EBN 75), (150 mg/kg BW) (EBN 150), and glyburide (GR) for an eight-week study. H&E and Masson's trichrome staining were utilized to investigate the retinal tissue and vascular changes. The immunofluorescence study was used to detect angiogenic protein expression. The vascular corrosion cast/SEM method was also used to evaluate capillary plexus formation within the retinal layer. From histological studies, DM rats have thinning of the retinal layer. Remarkably, the retinal vessels displayed dilations resembling ruptured blood vessels. The expression of vascular endothelial growth factor (VEGF) (30.51±2.62), cluster of differentiation 31 (CD31) (28.18±0.22), and platelet-derived growth factor receptor beta (PDGFR-β) (141.67±0.97) were increased. EBN 75 exhibited some small improvements in their blood vessels and eye tissue. At a dose of 150 mg/kg BW, EBN proved to be more effective. There was a significant decrease in VEGF and CD31 expression compared with the diabetic group (p<0.001 and p<0.01, respectively). These studies have demonstrated that EBN can lower the growth levels of VEGF, CD31, and PDGFR-β, which results in a decrease in angiogenesis and a recovery from a variety of diabetic retinopathy-related diseases.

Pharmacokinetic and Pharmacodynamic Interactions of Momordica charantia extract with Glibenclamide in Streptozotocin induced Diabetic rats

Diabetes mellitus (DM) has emerged as a leading global health problem. Diabetes is estimated to affect 425 million adults globally. Majority of diabetic patients take herbal medicines as supplement for the treatment of diabetes. The combined use of herbs and modern synthetic medicine has increased the possibility of pharmacokinetic (PK) and pharmacodynamic (PD) interactions. Momordica charantia (MC) commonly known as bitter melon/bitter gourd, is globally marketed for treatment of diabetes. Many diabetic patients consume MC extract along with antidiabetic drugs for better therapeutic effect with or without knowledge of health care provider.The current study evaluated PKPD herb-interaction of MC extract with glibenclamide (GLB) following oral administration either alone or in combination with MC extract for 28-consequative days. MC extract co-administratioin showed increased plasma exposure of GLB on Day-28 compared to glibenclamide alone treated groups with similar dosing regimen. Glibenclamide upon co-administration with MC extract showed improved pharmacodynamic parameters compared to glibenclamide alone treated groups suggesting synergistic or additive effect. Physician can reduce the glibenclamide dose in diabetic patients if they are taking MC as supplement.

Equilibrium solubility and mathematical modelling of glibenclamide in cosolvent mixtures of glycols and alcohols

ABSTRACT Glibenclamide (GBN) is a commonly used oral medication for treating type-II diabetes that acts by reducing high blood sugar levels. The effectiveness of GBN depends on the quality of the product. The quality is influenced by the solubility properties of a drug. This study aimed to measure the solubility of GBN in various solvent mixtures including some glycols and alcohols. The solubility was determined using a shake-flask method at 298.2 K under atmospheric pressure. The results showed that GBN had the highest solubility in ethylene glycol monomethyl ether and alcohol mixtures compared to the other tested glycols.

The antioxidant, antidiabetic, and antihyperlipidemic effects of the polyphenolic extract from Salvia blancoana subsp. mesatlantica on induced diabetes in rats

Currently, several studies have demonstrated the benefits of medicinal plants in managing type 2 diabetes. In this work, we evaluated the beneficial effects of the polyphenolic extract (PESB) from Salvia blancoana subsp. mesatlantica in the management of hypercaloric-feeding and small-dose alloxan-brought type 2 diabetes in rats. We analyzed the chemical constituents of the extract, including flavones and flavonols content, to understand its biological action. The antioxidant activities were evaluated by total antioxidant action, scavenging effect of the free radical DPPH, and reducing power. The obtained results showed that the value of TFC was estimated at 31.90 ± 0.34 mgEQ/g in the PESB extract. The total antioxidant capacity was estimated at 593.51 ± 4.09 mg (EAA)/g, the value of DPPH IC50 was 7.3 ± 0.00 μg/mL, and the value of EC50 of reducing power was estimated at 6.43 ± 0.01 μg/mL. In total, 14 phenolic compounds were identified and the naringin was the most dominant (63.19%) while the vanillin was the less recorded (0.10%). Serum glucose decreased significantly (p < 0.05) in rats given PESB (100 mg/kg) after four weeks. Glibenclamide (GLB) and PESB reduced HbA1c and increased plasma insulin in diabetic rats, restoring HOMA-β and HOMA-IR levels to near-normal. Additionally, diabetic rats treated with GLB or PESB showed statistically equivalent results to those of non-diabetic rats regarding hepatic enzymes, renal and lipid markers, as well as cardiovascular indices. The weight loss was significantly lower in diabetic rats receiving a dose of PESB (100 mg/kg), and GLB compared to corresponding untreated diabetic rats (p < 0.01). PESB and GLB showed a prominent protective function in the pancreas, liver, and kidney tissues. This investigation demonstrates the capacity of extracts from leaves of S. blancoana subsp. mesatlantica to manage diabetes mellitus due to their richness in a wide range of bioactive compounds. Therefore, more investigations are required to estimate the safety of the plant use.Graphical

Quality-by-design optimized RP-HPLC approach for the therapeutic drug monitoring of glibenclamide and fluoxetine in human plasma

Glibenclamide (GLB) and fluoxetine (FLX) were reported to have a potential drug-drug interaction when clinically co-administered. FLX is an inhibitor of CYP2C9 (the enzyme responsible for the metabolism of GLB), thus resulting in increased serum levels of GLB followed by severe hypoglycemia. Therefore, a sensitive, simple, and selective HPLC method is urgently needed for their simultaneous determination in plasma to be utilized for monitoring their therapeutic blood concentration levels. The current study introduces the first RP-HPLC approach for the simultaneous estimation of this binary mixture in less than 7 min. Adopting a quality-by-design (QbD) approach allowed for the designed approach to be optimized. Using a two-level full factorial design (25), the effects of various variables on the chromatographic responses were investigated and optimized. Hypersil BDS C18 column was used for the achievement of the optimum chromatographic separation at 19.9 °C using acetonitrile: sodium dihydrogen phosphate buffer (7.5 mM; pH 4, 60:40 % v/v) as a mobile phase, pumped isocratically at a flow rate of 1.036 mL/min. An injection volume of 30.0 μL was used for the binary mixture with UV detection at 230 nm. FLX and GLB calibration curves revealed an excellent linearity over the ranges of 0.025–10.00 and 0.02–10.00 µg/mL with limits of quantitation of 0.018 and 0.02 µg/mL, respectively. Due to the high sensitivity and selectivity, the method was successfully applied to the assay of the two drugs in spiked human plasma samples with reasonable % recoveries (92.70–108.50 %), indicating good bioanalytical applicability. Therefore, this study could be applied for the therapeutic drug monitoring of GLB and FLX in human plasma to control their potential drug-drug interaction. The suggested approach was also applied to estimate the investigated drugs in their dosage forms with reasonable % recoveries (96.80–102.80 %) and small % RSD values (less than 2 %). The method's excellent greenness was evaluated using three metric tools, including Analytical Eco-Scale, Green Analytical Procedure Index (GAPI), and Analytical Greenness (AGREE). Validation of the proposed method was performed according to ICH and US FDA guidelines.

Pharmacogenetics: Influence of CYP2C9*2 and *3 alleles polymorphisms on Iraqi type 2 diabetic patients

Abstract Background: Sulfonylureas (SUs) are the most prescribed anti-diabetic drugs. The enzyme responsible for metabolizing of SUs is hepatic cytochrome P4502C9 (CYP2C9). The CYP2C9 gene has numerous allelic variations; among those, the CYP2C9*2 and CYP2C9*3 are the most common and clinically significant allelic variations. The pharmacokinetics of SUs are dramatically impacted by CYP2C9 genotype. Objective: Evaluation of the association of genetic polymorphisms in CYP2C9 gene with the efficacy of glibenclamide (GB), second-generation SUs, by investigating two CYP2C9 allelic variants. Materials and Methods: Blood samples were collected from 113 type 2 diabetes patients. Allele specific amplification-polymerase chain reaction was used to genotype the CYP2C9 gene. Fasting serum glucose, fasting insulin, and glycated hemoglobin (HbA1c) levels were measured as part of the biochemical analysis. Results: The CYP2C9 gene variants were analyzed in a study group. The results exposed that 75 patients carried the wild (CYP2C9*1/*1) genotype, 25 were heterozygote allele (CYP2C9*1/*2) for CYP2C9*2 gene, 4 were homozygous for the variant CYP2C9*2 allele (CYP2C9*2/*2), and 9 were heterozygous for the variant CYP2C9*3 allele (CYP2C9*1/*3). Statistically significant difference was found in mean HbA1c between the mutant and wild alleles group (P = 0.044). The mean HbA1c for those carrying the CYP2C9*2 and*3 alleles (n = 38) was 8.4750 compared to 9.3177 for those carrying the CYP2C9*1 allele (n = 75), which indicate better glycemic control. Conclusion: The accordance of CYP2C9*2 and*3 was found to be associated with severe hypoglycemia (odd ratio [OR] = 2.045). The OR suggests a strong association between CYP2C9*2 and*3 alleles and hypoglycemia. Our findings imply that the diabetic patients with CYP2C9 polymorphism are more likely to suffer hypoglycemia than those with wild type alleles when treated with GB.

Antispasmodic and antidiarrheal effects of Juniperus oxycedrus L. on the jejunum in rodents.

Functional bowel disorders (FBD) have a major potential to degrade the standards of public life. Juniperus oxycedrus L. (J. oxycedrus) (Cupressaceae) has been described as a plant used in traditional medicine as an antidiarrheal medication. The present study is the first to obtain information on the antispasmodic and antidiarrheic effects of J. oxycedrus aqueous extract through in vitro and in vivo studies. An aqueous extract of J. oxycedrus (AEJO) was extracted by decoctioning air-dried aerial sections of the plant. Antispasmodic activity was tested in an isolated jejunum segment of rats exposed to cumulative doses of drogue extract. The antidiarrheic activity was tested using diarrhea caused by castor oil, a transit study of the small intestine, and castor oil-induced enteropooling assays in mice. In the jejunum of rats, the AEJO (0.1, 0.3 and 1 mg/ml) diminished the maximum tone induced by low K+ (25 mM), while it exhibited a weak inhibitory effect on high K+ (75 mM) with an IC50=0.49 ± 0.01 mg/ml and IC50=2.65 ± 0.16 mg/ml, respectively. In the contractions induced by CCh (10-6 M), AEJO diminished the maximum tone, similar to that induced by low K+ (25 mM). with an IC50=0.45 ± 0.02 mg/ml. The inhibitory effect of AEJO on low K+ induced contractions was significantly diminished in the presence of glibenclamide (GB) (0.3 µM) and 4-aminopyrimidine (4-AP) (100 µM), with IC50 values of 1.84 ± 0.09 mg/ml. and 1.63 ± 0.16 mg/ml, respectively). The demonstrated inhibitory effect was similar to that produced by a non-competitive antagonist acting on cholinergic receptors and calcium channels. In castor oil-induced diarrhea in mice, AEJO (100, 200, and 400 mg/kg) caused an extension of the latency time, a reduced defecation frequency, and a decrease in the amount of wet feces compared to the untreated group (distilled water). Moreover, it showed a significant anti-motility effect and reduced the amount of fluid accumulated in the intestinal lumen at all tested doses. These findings support the conventional use of Juniperus oxycedrus L. as a remedy for gastrointestinal diseases.

Dispersive micro solid phase extraction of glibenclamide from plasma, urine, and wastewater using a magnetic molecularly imprinted polymer followed by its determination by a high-performance liquid chromatography-photodiode array detector.

The present study describes the development of a simple and selective analytical method for dispersive micro solid phase extraction and determination of glibenclamide (GLB) using magnetic molecularly imprinted polymer (MMIP) as a sorbent. MMIP was fabricated by the non-covalent method on the surface of silicated Fe3O4 and had a high affinity for glibenclamide; dual monomers, itaconic acid and allylamine, were used for this. Polymerization was achieved by the precipitation method in the presence of glibenclamide as the template and ethylene glycol dimethacrylate as the cross-linker. The morphology and structural properties of the MMIP were characterized by different analytical methods. To achieve maximum extraction efficiency, influencing parameters were optimized. The linearity range was 1-2000 and 12-2000 μg L-1 by high-performance liquid chromatography-photodiode array detector (HPLC-PDA) and UV-vis spectroscopy, respectively. The detection and quantification limits with UV-vis and HPLC-PDA analyses were 4 and 12 μg L-1 and 0.3 and 1 μg L-1, respectively. Under optimized conditions, recovery of glibenclamide spiked in plasma, human urine, and wastewater was between 89.4 and 102.9% at the concentration levels of 25, 250, and 500 μg L-1; relative standard deviations were below 3.7% by HPLC-PDA. The developed method has a favorable pre-concentration factor of 140.0. Equilibrium data and sorption isotherms fitted well with the Langmuir model. A maximum sorption capacity of 24.260 mg g-1 was acquired based on the Langmuir model. The synthesized sorbent with high selectivity was used to separate GLB from complex biological systems and wastewater before measurement with UV-vis or HPLC-PDA.

Successful Prediction of Human Hepatic Concentrations of Transported Drugs Using the Proteomics-Informed Relative Expression Factor Approach.

Tissue drug concentrations determine the efficacy and toxicity of drugs. When a drug is the substrate of transporters that are present at the blood:tissue barrier, the steady-state unbound tissue drug concentrations cannot be predicted from their corresponding plasma concentrations. To accurately predict transporter-modulated tissue drug concentrations, all clearances (CLs) mediating the drug's entry and exit (including metabolism) from the tissue must be accurately predicted. Because primary cells of most tissues are not available, we have proposed an alternative approach to predict such CLs, that is the use of transporter-expressing cells/vesicles (TECs/TEVs) and relative expression factor (REF). The REF represents the abundance of the relevant transporters in the tissue vs. in the TECs/TEVs. Here, we determined the transporter-based intrinsic CL of glyburide (GLB) and pitavastatin (PTV) in OATP1B1, OATP1B3, OATP2B1, and NTCP-expressing cells and MRP3-, BCRP-, P-gp-, and MRP2-expressing vesicles and scaled these CLs to in vivo using REF. These predictions fell within a priori set twofold range of the hepatobiliary CLs of GLB and PTV, estimated from their hepatic positron emission tomography imaging data: 272.3 and 607.8 mL/min for in vivo hepatic sinusoidal uptake CL, 47.8 and 17.4 mL/min for sinusoidal efflux CL, and 0 and 4.20 mL/min for biliary efflux CL, respectively. Moreover, their predicted hepatic concentrations (area under the hepatic concentration-time curve (AUC) and maximum plasma concentration (Cmax )), fell within twofold of their mean observed data. These data, together with our previous findings, confirm that the REF approach can successfully predict transporter-based drug CLs and tissue concentrations to enhance success in drug development.

Optimization and Validation of Sensitive UPLC-PDA Method for Simultaneous Determination of Thymoquinone and Glibenclamide in SNEDDs Formulations Using Response Surface Methodology

The development of analytical procedures capable of simultaneous determination of two or more drugs is in crucial demand due to the availability of different formulations that are composed of different APIs. The presented study aimed to optimize and validate a simple, accurate, and sensitive UPLC analytical method for the simultaneous determination of thymoquinone (TQ) and Glibenclamide (GB) using response surface methodology, and apply this method in pharmaceutical formulations. A 32 full design of experiment was utilized to study the impacts of the independent parameters (acetonitrile ACN concentration, A; and column temperature, B) on the drugs’ analytical attributes (viz, retention time, peak area, and peak asymmetry, in addition to the resolution between TQ and GB peaks). The results revealed that the independent parameters exhibited highly significant (p &lt; 0.05) antagonistic effects on retention times for TQ and GB peaks, in addition to the agnostic effect on GB peak symmetry (p-value = 0.001). Moreover, antagonistic impacts (p &lt; 0.05) on the resolution between TQ and GB peaks were found for both independent factors (A and B). The statistical software suggested 46.86% of ACN (A) and 38.80 °C for column temperature (B) for optimum analytical responses. The optimized green method was discovered to be acceptable in terms of selectivity, precision, accuracy, robustness, sensitivity, and specificity. Moreover, the optimized simultaneous method was successfully able to determine the contents of TQ and GB in self-nanoemulsifying drug delivery (SNEDD) formulation, in which the results showed that GB and TQ content within the prepared formulations were 1.54 ± 0.023 and 3.62 ± 0.031 mg/gm, respectively. In conclusion, the developed assay was efficient and valid in analyzing TQ and GB simultaneously in bulk and self-nanoemulsifying drug delivery system (SNEDDs) formulations.

Investigation of drug-polymer miscibility and design of ternary solid dispersions for oral bioavailability enhancement by Hot Melt Extrusion

The present investigation aimed to enhance the oral bioavailability of the poorly water-s Glibenclamide (GLB) using amorphous solid dispersions (ASD) with various polymers. The drug-polymer miscibility was predicted using in-silico molecular dynamics simulation, Hansen solubility parameter, Flory-Huggins theory, and Gibb’s free energy calculation. To enhance the solubility further, the effervescence amorphous solid dispersions (ESD) technique was included. The solid dispersions (SDs) were initially prepared using the solvent evaporation method. The polymers that enhanced the solubility were optimized by the Design of Experiments (DoE) approach and Hot Melt Extruder (HME) technique. The optimized formulation were subjected to various solid and liquid state characterizations. The XRD and DSC study confirmed conversion into the amorphous form, and hydrogen bonding between the drug and polymer was confirmed by FTIR and 1H NMR. The morphology was assessed by SEM study. The optimized formulation showed enhanced solubility and dissolution data. The optimized formulation remained stable at the accelerated conditions for six months. The optimized formulation subjected to the pre-clinical pharmacokinetic and pharmacodynamics study showed a better pharmacokinetics profile of the drug and enhanced pharmacological activity. Introducing the effervescent technique to the conventional ASD technique helped to enhance the physicochemical properties with improved pharmacological action.

Formulation of Glibenclamide proniosomes for oral administration: Pharmaceutical and pharmacodynamics evaluation

Glibenclamide (GB), oral antidiabetic sulfonylurea, is used in the management of diabetes mellitus type II. It suffers from low bioavailability due to low water solubility. This work aimed to enhance the dissolution of GB by formulating the drug as a proniosomes which then improves the pharmacological effect. GB proniosomal formulations were prepared using a slurry method with sucrose as a carrier. The formulations were characterized by particle size, zeta potential, entrapment efficiency %, flow properties of the powder, and in vitro dissolution study. The pharmacological effect was also assessed by determining and measuring the fasting blood glucose level (BGL) before and after the treatment. Formulating GB proniosomes with the slurry method produces a free-flowing powder with a particle size range from 190.050 ± 43.204 to 1369.333 ± 150.407 nm and the zeta potential was above 20 mV (-24 to −58 mV), indicating good stability. The dissolution rate for all formulations was higher than that of the pure drug, indicating the efficiency of the proniosome in enhancing the drug solubility. A significant reduction in the fasting blood glucose level (73 %) was observed in animals treated with proniosomal formulation with no sign of liver damage. In contrast, the pharmacodynamics results show a significant reduction in fasting blood glucose level for animals treated with proniosomes compared to a 17.6 % reduction in BGL after treatment with pure drug. Moreover, the histopathological results showed no sign of liver damage that occurred with proniosomal treatment. GB proniosomal formulations is a promising drug delivery system with good therapeutic efficacy and stability.

Validation and development of RP-HPLC method for quantification of glibenclamide in rat plasma and its application to pharmacokinetic studies in wistar rats

Herein, we report a novel, simple, specific, accurate and cost-friendly validated reverse phase-high performance liquid chromatographic (RP-HPLC) method for the quantification of second generation sulphonylurea based antidiabetic drug, glibenclamide (GLB) in rat plasma and its application to calculate pharmacokinetic parameters in wistar rats. The internal standard used was flufenamic acid. The chromatographic separation was conducted on C18 column (250 mm × 4.6 mm x 5 μm, Agilent-Zorbax, SB) using isocratic elution with mobile phase containing Acetonitrile: Water (1:1; v/v) pH adjusted to 4.0 with 0.03 % glacial acetic acid and detected by photo-diode array as detector. Calibration curves made in the rat plasma were linear in the range of 50–1200 ng/ml with r2 = 0.998. The LLOQ was 40 ng/ml. This method was effectively applied for pharmacokinetic studies of Glibenclamide following administration through oral route as solid dispersion formulation to Wistar rats. Several methods are available in the literature which can be employed for the quantification of Glibenclamide but such methods are tedious, provide lower sensitivity, less simultaneous resolution and are time-consuming. Therefore the present methods suits best for the quantification of Glibenclamide from Wistar rats.

A systematic review and meta-analysis on the efficacy of glibenclamide in animal models of intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) is a devastating stroke with many mechanisms of injury. Edema worsens outcome and can lead to mortality after ICH. Glibenclamide (GLC), a sulfonylurea 1- transient receptor potential melastatin 4 (Sur1-Trpm4) channel blocker, has been shown to attenuate edema in ischemic stroke models, raising the possibility of benefit in ICH. This meta-analysis synthesizes current pre-clinical (rodent) literature regarding the efficacy of post-ICH GLC administration (vs. vehicle controls) on behaviour (i.e., neurological deficit, motor, and memory outcomes), edema, hematoma volume, and injury volume. Six studies (5 in rats and 1 in mice) were included in our meta-analysis (PROSPERO registration = CRD42021283614). GLC significantly improved behaviour (standardized mean difference (SMD) = -0.63, [-1.16, -0.09], n = 70-74) and reduced edema (SMD = -0.91, [-1.64, -0.18], n = 70), but did not affect hematoma volume (SMD = 0.0788, [-0.5631, 0.7207], n = 18-20), or injury volume (SMD = 0.2892, [-0.4950, 1.0734], n = 24). However, these results should be interpreted cautiously. Findings were conflicted with 2 negative and 4 positive reports, and Egger regressions indicated missing negative edema data (p = 0.0001), and possible missing negative behavioural data (p = 0.0766). Experimental quality assessed via the SYRCLE and CAMARADES checklists was concerning, as most studies demonstrated high risks of bias. Studies were generally low-powered (e.g., average n = 14.4 for behaviour), and future studies should employ sample sizes of 41 to detect our observed effect size in behaviour and 33 to detect our observed effect in edema. Overall, missing negative studies, low study quality, high risk of bias, and incomplete attention to key recommendations (e.g., investigating female, aged, and co-morbid animals) suggest that further high-powered confirmatory studies are needed before conclusive statements about GLC's efficacy in ICH can be made, and before further clinical trials are performed.

Sevoflurane-mediated modulation of oxidative myocardial injury.

Volatile anesthetics offer protection when administered throughout an ischemic injury. We examined how volatile anesthetics modulate the cardiac myocytic injury associated with hydrogen peroxide. Forty-eight Long-Evans rats were divided into four groups depending on the treatment: none (CONT), Glibenclamide (GLB); Sevoflurane (SEV); or GLB+SEV. Each group was further divided into two, one of which was exposed to hydrogen peroxide (H2O2). Oral GLB was administered 48 hours before myocardial isolation. All rats were anesthetized by intraperitoneal injection of Ketamine, and the hearts were harvested after heparinization. Cardiomyocytes were isolated using a combination of mechanical mincing and enzymatic digestion. After isolation, the aliquots of cells were exposed to H2O2 and FeSO4 for 30 minutes. The cell suspensions were then bubbled for 10 minutes with 100% oxygen and 1.5% SEV if appropriate. Apoptosis was detected by fluorescein-bound annexin-V (ANX-V), necrosis by propidium iodide, and ELISA assessed caspase-3 activity in all groups. There was an increase in apoptosis, necrosis, and caspase-3 activity in the cells following exposure to hydrogen peroxide. SEV reduced the rate of cell necrosis and apoptosis. Pretreatment with GLB did not alter the effects of SEV. Similarly, caspase-3 activity did not change with GLB, although SEV administration reduced this enzymatic activity in response to hydrogen peroxide. In this oxidant injury model, we demonstrated that incubating isolated cardiomyocytes with SEV profoundly diminished H2O2-induced apoptotic and necrotic cells compared to their CONTs. These results support the hypothesis that KATP channels are not the sole mediators associated with anesthetic preconditioning.