Drug Diazepam Research Articles

General anesthesia activates a central anxiolytic center in the BNST

Low doses of general anesthetics like ketamine and dexmedetomidine have anxiolytic properties independent of their sedative effects, but the underlying mechanisms remain unclear. We discovered a population of GABAergic neurons in the oval division of the bed nucleus of the stria terminalis that are activated by multiple anesthetics and the anxiolytic drug diazepam (ovBNSTGA). The majority of ovBNSTGA neurons express neurotensin receptor 1 (Ntsr1) and form circuits with brain regions known to regulate anxiety and stress responses. Optogenetic activation of ovBNSTGA or ovBNSTNtsr1 neurons significantly attenuated anxiety-like behaviors in both naive animals and mice with inflammatory pain, while inhibition of these cells elevated anxiety. Activation of these neurons decreased heart rate and increased heart rate variability, suggesting that they reduce anxiety by modulating autonomic responses. Our study identifies ovBNSTGA/ovBNSTNtsr1 neurons as a common neural substrate mediating the anxiolytic effect of low-dose anesthetics and a potential therapeutic target for treating anxiety-related disorders.

Anxiolytic-like effect of daidzin possibly through GABAA receptor α2 and α3 subunits interaction pathway: In vivo and in silico studies

The soy plant-derived isoflavone daidzin (DZN) has diverse biological activities, including neuroprotective (e.g., memory-enhancing and antiepileptic) effects. This study emphasizes evaluating the anxiolytic effect of DZN on mice. Additionally, in silico investigations were also carried out to check the potential molecular mechanisms for the anxiolytic effect of DZN. For this, adult male Swiss albino mice were intraperitoneally (i.p.) treated with DZN (5, 10, and 20 mg/kg) with or without the standard GABAergic agonist drug diazepam (DZP: 2 mg/kg) and/or antagonist drug flumazenil (FLU: 0.1 mg/kg) and checked for different locomotor behaviors using various mouse models. The molecular docking study of DZN was conducted against GABAA receptor subunits. Findings suggest that DZN dose-dependently and significantly (p <0.05) increased locomotor activities such as the number of field crosses, hole crosses, swings, grooming, and light residence time while decreasing the rearing of the animals. With DZP, it significantly (p <0.05) reduced the test parameters, while altering these parameters with FLU. Our molecular docking studies demonstrate that DZN has a strong binding affinity of −7.4 and −6.9 kcal/mol for α2 and α3 subunits of the GABAA receptor, respectively, whereas the standard drugs DZP and FLU showed binding affinities between −6.0 and −6.7 kcal/mol for these subunits. Taken together, DZN augmented the anxiolytic effect of DZP while reducing the effect of FLU in mice. We suppose that DZN may show anxiolytic-like effects on Swiss mice, possibly through α2 and α3 subunits of the GABAA receptor interaction pathway.

Evaluation of Anthelmintic and Neuropharmacological Activities of the leaves of Chassalia

Chassalia curviflora belongs to the family Rubiaceae has been documented for its anticancer, anthelmintic &amp; neuropharmacological effects. This study was performed to gauge the anticancer, anthelmintic and neuropharmacological activities of the methanolic extract from the leaves of Chassalia curviflora (MECC). Anthelmintic activity was investigated using earthworms (Pheretima posthuma) and fresh leaf juice of the plant. Anticancer test was done using cell viability assay. During anticancer test in the concentration of 1000µg/mL the percent of inhibition was 29.16%. In anthelmintic activity, the time of paralysis for fresh leaf juice was begin from 8 min 19 s and end to 38 min 21 s while the time for tested drug Albendazole was started from 9 min 19 s and end to 46 min 52 s. Besides, the time of death start from 10 min 35 s and end to 63 min 19 s for leaf juice and for Albendazole the time of death start from 22 min 36 s and end to 75 min 3 s respectively at several doses which suggest considerable anthelmintic activity of the plant. Neuropharmacological activity was performed by hole cross and light/dark box tests at the doses of 200 mg/kg and 400 mg/kg in Swiss albino mice contrast to the test drug Diazepam (2mg/kg). The extract significantly decreases the locomotor activity as shown by the results of hole cross test which indicate significant antidepressant property. The results also matching the activity in light/dark box test. Thus, the obtained results in the research work provide a support for the utilization of this plant for medicinal purposes and encourage further investigations for more fruitful results.

Effects of acute administration of 4-allyl-2,6-dimethoxyphenol in mouse models of seizures

Epilepsy, a chronic neurological disorder characterized by recurrent unprovoked seizures, presents a substantial challenge in approximately one-third of cases exhibiting resistance to conventional pharmacological treatments. This study investigated the effect of 4-allyl-2,6-dimethoxyphenol, a phenolic compound derived from various natural sources, in different models of induced seizures and its impact on animal electroencephalographic (EEG) recordings. Adult male Swiss albino mice were pre-treated (i.p.) with a dose curve of 4-allyl-2,6-dimethoxyphenol (50, 100, or 200 mg/kg), its vehicle (Tween), or standard antiepileptic drug (Diazepam; or Phenytoin). Subsequently, the mice were subjected to different seizure-inducing models – pentylenetetrazole (PTZ), 3-mercaptopropionic acid (3-MPA), pilocarpine (PILO), or maximal electroshock seizure (MES). EEG analysis was performed on other animals surgically implanted with electrodes to evaluate brain activity. Significant results revealed that animals treated with 4-allyl-2,6-dimethoxyphenol exhibited increased latency to the first myoclonic jerk in the PTZ and PILO models; prolonged latency to the first tonic-clonic seizure in the PTZ, 3-MPA, and PILO models; reduced total duration of tonic-clonic seizures in the PTZ and PILO models; decreased intensity of convulsive seizures in the PTZ and 3-MPA models; and diminished mortality in the 3-MPA, PILO, and MES models. EEG analysis indicated an increase in the percentage of total power attributed to beta waves following 4-allyl-2,6-dimethoxyphenol administration. Notably, the substance protected from behavioral and electrographic seizures in the PTZ model, preventing increases in the average amplitude of recording signals while also inducing an increase in the participation of theta and gamma waves. These findings suggest promising outcomes for the tested phenolic compound across diverse pre-clinical seizure models, highlighting the need for further comprehensive studies to elucidate its underlying mechanisms and validate its clinical relevance in epilepsy management.

Ovarian Gene Transcriptional Responses to Anticonvulsant Drugs (Diazepam and Phenytoin) in Female Wistar Rats

This study was designed to investigate ovarian gene transcriptional responses to selected anticonvulsant drugs (diazepam and phenytoin) in female rats. Fifteen female rats (120 – 140 g) were used for this study. Diazepam (0.14 mg/kg) and phenytoin (2.8 mg/kg) were given to the rodents orally for 50 days. The method of RT-PCR involving isolation of RNA, conversion of cDNA and electrophoresis was employed to investigate the expression of FSH-R, aromatase and GPX-1genes. Graphics were generated as mean +/- SEM using Graph-pad Prism version 8.0. The expression of GPX-1 was significantly (p&lt;0.05) up-regulated, while the FSH-R and aromatase expressions were significantly (p&lt;0.05) and insignificantly (p&gt;0.05) down-regulated respectively in the diazepam treated rats when compared with their respective controls. The FSH-R and aromatase expressions were significantly (p&lt;0.05) up-regulated, while the GPX-1 expression was insignificantly (p&gt;0.05) up-regulated in the phenytoin treated rats relative to their respective controls. Conclusively, it can be suggested that diazepam: inhibited follicular growth through the down-regulation of FSH-R expression, reduced estrogen level through the down-regulation of aromatase expression, inhibited the production of reactive oxygen species and oxidative stress through the up-regulation of GPX-1 expression. In addition, it can be suggested that phenytoin: induced follicular growth through the up-regulation of FSH-R expression, increased estrogen level through the up-regulation of aromatase expression, inhibited the production of reactive oxygen species and oxidative stress through the up-regulation of GPX-1 expression.

Effects of chronic administration and withdrawal of phlorotannin supplement on sleep-wake profiles in mice: A comparative study with the hypnotic drug diazepam

The aim of this study was to investigate the effects of chronic administration (21 days) of phlorotannin supplement (PS) on sleep and compare them with the effects of diazepam (DZP), a benzodiazepine hypnotic drug. DZP significantly decreased sleep latency and increased non-rapid eye movement sleep amount on day 1 of administration compared with those at baseline; however, its effects diminished over the administration period. On the contrary, PS showed significant sleep-promoting effects throughout the study period, with no significant difference in efficacy between days 1 and 21. Notably, PS did not cause any changes in the slow wave activity (SWA) throughout the treatment period; whereas, DZP decreased the SWA on day 1 of administration. These results suggest that chronic administration of PS does not induce tolerance, implying its consistent sleep-maintaining effects.

Benzodiazepine derivatives for the treatment of neuropharmacological disorders and pain management: Docking investigations and in-vivo studies

Benzodiazepines are well-known for their medicinal properties. The current study aimed to evaluate and seek new therapeutic possibilities of some of these compounds for addressing anxiety and pain. Four benzodiazepine derivatives were synthesized, characterized, and subjected to computational assessment of the GABAergic response upon interaction with human GABA-A receptors (PDBID: 6×3x) relative to the drug diazepam (DZP) to achieve the objective. Apart from the docking analysis, ADME analysis of the compounds was also performed. Following the in-silico evaluations, two of the synthesized compounds (designated as P3 and P4) were identified for subsequent in-vivo investigations utilizing laboratory rats to explore their anxiolytic potential in comparison to diazepam, and their analgesic efficacy relative to diclofenac sodium. The combination of in-silico and in-vivo assessments uncovered the binding sites of P4 (a fluoro derivative) and its potential as a superior anxiolytic drug compared to P3 (a hydroxy derivative) for analgesic relief. The ADME pharmacokinetic evaluation of the synthesized compounds by Lipinski rule of 5 suggested the oral bioavailability of these drugs. These findings are important as they provide valuable insights into the potential development of new anxiolytic and analgesic agents from the benzodiazepine derivatives that can be easily synthesized and developed into affordable drugs.

Leaching of plasticizers from PVC medical devices: A molecular interpretation of experimental migration data

High pressure liquid chromatography (HPLC) experiments and molecular simulations were combined to investigate the leaching process of DINCH, DEHT and TOTM plasticizers from PVC materials. Methodologies based on statistical thermodynamics were used to rationalize the migration of the different plasticizers in water and ethanolic solutions from different free energy contributions. We extended this combined approach to the study of the adsorption of a model drug (diazepam) on different plasticized-PVC materials. The drug loss by sorption was investigated in different solutions and rationalized at an energy level. The combination of experiments and theoretical approaches proved to be effective for these sophisticated systems and processes for which various energy contributions are involved.

A comparative study of anti-anxiety properties of ethanolic and aqueous extracts of Ocimum sanctum in animal models

Background: Anxiety transcends a natural emotion, evolving into a pathological condition with the potential to trigger cascading cardiovascular and psychiatric disorders. Although conventional allopathic medicine offers treatment options, concerns regarding their side effects and long-term efficacy remain prevalent. Medicinal plants contain natural compounds that may be promising sources of therapeutic drugs. Aims and Objectives: This study aims to compare the anxiolytic potential of ethanolic and aqueous extracts of Ocimum sanctum (OS). The growing appeal of plant-based therapies for anxiety stems from perceived advantages in safety and tolerability compared to synthetic drugs. Materials and Methods: The anxiolytic activity of aqueous and ethanolic extract of OS is evaluated with an elevated plus maze test. A total of 36 Wistar albino rats (150–200 g) were used and randomly divided into six groups of six animals each. The effects of the test drug at different doses, 100 and 200 mg/kg, were compared with the standard anxiolytic drug diazepam at 2 mg/kg body weight and the control group using distilled water at 0.5 mL/kg body weight. Results: The behavioral changes suggested reduced anxiety and openarm exploration in plus-maze indicates reduced anxiety in animals treated with OS extracts. The changes are significantly (P &lt; 0.001) comparable with the standard drug diazepam. Conclusion: The ethanolic extracts of OS are more significant than aqueous extracts for evaluating anti-anxiety activity in a dose-dependent manner. In addition, ethanolic extracts are more likely to contain a wider range of bioactive compounds, which are thought to be responsible for the anti-anxiety effects of OS.

Exploiting the synergistic effects of combined carbon‐based materials for rape drug diazepam electrochemical sensing: Application to simulated casework matrices

AbstractDiazepam (DZP), a long‐lasting benzodiazepine and controlled substance, is prevalent on the illegal market, notably as a date rape drug. This study presents a novel electrochemical sensor (CPE/GCPE/MWCNTs) for DZP determination in lab routines. Characterized through LSV, EIS, and SEM, the sensor revealed a pH‐dependent, irreversible reduction process at approximately −0.70 V (vs. Ag/AgCl(sat)). Optimization with SWV and 0.1 mol L−1 B–R solution (pH 1.0) produced a calibration curve spanning 1.49 to 21.5 μmol L−1. LOD and LOQ were 0.33 and 1.10 μmol L−1, respectively. Quantification in spiked samples exhibited no significant differences compared to HPLC‐DAD, endorsing the sensor for monitoring date rape drugs in forensic analysis.

Study of Anticonvulsant effect of Simvastatin in Maximal Electroshock and Pentylenetetrazole Induced Seizure Model In Albino Mice

Epilepsy is a common neurological condition affecting people of all ages and both sexes. Most of the currently used antiepileptic drugs have neurotoxic and teratogenic effects. There are scanty reports of anticonvulsant activity with statins. The present study was undertaken to evaluate the anticonvulsant effect of simvastatin using MES and PTZ models in mice and compare it with the standard drugs phenytoin and diazepam respectively.64 albino mice of either sex weighing 25-35g were divided into 8 groups of 8 mice each – 4 group each for MES and PTZ model. In MES model, seizures were induced via trans auricular electrodes with a current of 80mA for 0.2 second. Each mouse was pretreated with drugs 30 minutes before in both models. The different groups 1 to 4 were administered polyethylene glycol (0.5ml/Kg), phenytoin (20mg/kg), simvastatin (10mg/Kg) and simvastatin (20mg/Kg) respectively. In PTZ model, seizures were induced by giving PTZ 80mg/Kg. The different groups 5 to 8 were administered polyethylene glycol (0.5ml/Kg), diazepam (2mg/kg), simvastatin (10mg/Kg) and simvastatin (20mg/Kg) respectively. Statistical analysis was doneby ANOVA followed by Tukey’s post hoc analysis for group wise comparison. Results indicated that the anticonvulsant effect of simvastatin (10 and 20mg/Kg) was comparable to phenytoin in the MES model. In the PTZ model, simvastatin (10mg/Kg) produced a reduction in seizure duration but did not provide seizure protection as compared to that of the diazepam. The present study demonstrated the anticonvulsant effect of simvastatin. Further studies may explore the potential of this as an add-on drug in patients with dyslipidemia and epilepsy.

Evaluation of antidepressant activity of Polyherbal methanolic extract of leaves of Zingiber officinale, Azadirachta indica, Curcuma longa and Hibiscus rosasinesis

Aim: The Present studies were evaluate antianxiety activity of methanolic composite leaves extract of (Zingiber officinale, Azadirachta indica, Curcuma longa and Hibiscus rosasinesis) on administration in rats. Method: Male albino Wistar rat were taken and divided into 5 groups and each group contain 4 rats. Diazepam (1mg/kg) and composite extract (50mg/kg, 75mg/kg and 150mg/kg) were suspended in water and administrate orally. In these studies vehicle/ drugs were given 30min before exposure to experimental model such as actophometer and rotarod. C.E increase locometry actvity and time taken by rat to falling from the rod in actophometer and rotarod. The activity of rat is compared with standard drugs diazepam (1mg/kg). Result: The studied shown that at dose of 50mg/kg, 75mg/kg and 150mg/kg composite extract posse’s significant antidepressant activity.

Moringa extract reverses pilocarpine-induced hippocampal sclerosis in rats with temporal lobe epilepsy

The horseradish tree “Moringa oleifera” is the most nutritious terrestrial plant around the globe. Although native to India, its fast growth and drought resistance ability enabled the plant to be cultivated worldwide. In the current study, we report on the isolation of a new phenolic methyl ester namely moringinine A (1) along with four other known compounds viz. caffeic acid (2), ferulic acid (3), 4-hydroxybenzonitrile (4), and 4-hydroxyphenyl acetic acid (5) from Moringa seeds. The later compound was first to be isolated from family Moringaceae. Compounds identification was guided by interplay of NMR and HR-ESI-MS analysis. Anti-epileptic studies conducted in vivo showed that the extract attenuates convulsions by suppressing stress–induced pro-inflammatory markers TNF-α, IL-1β, IL-6, and IFN-ɣ whereas upregulating the anti-inflammatory markers TGF-β and IL-10 in the hippocampal tissues of epileptic rats. The isolated compounds were subjected to computational studies through docking on lactate dehydrogenase A(LDH) and interleukin-6 (IL-6), where all showed binding modes and interaction energies comparable to those of the reference drug diazepam. ADME investigation revealed good pharmacokinetic and drug-likeness properties. These results show that Moringa oleifera seeds could potentially be used as adjuvant in the management of epilepsy.

Xerostomia yang diinduksi obat pada Pasien dengan gangguan kecemasan : Laporan Kasus

Saliva plays an important role in oral health because it helps taste, protects the mucosa and teeth, and aids the digestive process. Low salivary flow is known as xerostomia. Xerostomia is caused by various factors, one of which is the consumption of certain types of drugs such as drugs in patients with anxiety disorders. The purpose of writing this journal is to report cases of xerostomia experienced by patients with anxiety disorders. A male patient came with complaints of dry oral mucosa and discomfort for chewing, swallowing, and speaking. The patient is undergoing treatment for an anxiety disorder and is taking the drugs diazepam, fluoxetine, alprazolam, and clozapine. Measurement of the patient's salivary flow rate showed results of 0.8 ml/minute when stimulated and 0.1 ml/minute without stimulation. Patients with anxiety disorders who have to take antidepressant drugs for a long time are advised to maintain oral hygiene so that they can improve their quality of life.

A novel physiologically based pharmacokinetic model of rectal absorption, evaluated and verified using clinical data on 10 rectally administered drugs

We present a physiologically based pharmacokinetic (PBPK) model simulating systemic drug concentrations following administration to the human rectum. Rectum physiology is parameterized based on literature data. The model utilizes in vitro release (IVRT) profiles from which drug mass transfer through the rectal fluid and tissue and into the systemic circulation are predicted. Due to a lack of data, rectal fluid and tissue absorption parameters are predicted either from colon absorption, with modifications relevant to rectal physiology, or optimized. The PBPK model is evaluated by simulating 29 clinical studies for 10 drugs. For 8 drugs (diazepam, diclofenac, indomethacin, naproxen, paracetamol, pentobarbital, phenobarbital and theophylline) the bias (average fold error, AFE) and precision (absolute average fold error, AAFE) of Cmax, AUC0-t and AUC0-inf simulations range from 0.87 to 2.22, indicating good agreement with observed values. For prochlorperazine and promethazine, the AFEs and AAFEs of Cmax predictions are 1.30 and 2.52, respectively. TheAUC0-t and AUC0-inf are overpredicted for both compounds(AFEs and AAFEs from 2.66 to 4.90). This results from a lack of reliable elimination data for prochlorperazine and the relevance of the IVRT profiles used in the promethazine model. The model paves the way for more mechanistic rectal drug absorption studies and virtual bioequivalence methods for rectal drug products.

Analysis of injectable drugs (Pentazocine and Diazepam) sampled from shops and companies in Anambra State, Nigeria

The incidences of drug adulteration and drug resistance have been a major problem facing mankind in recent years. Low and medium-income countries like Nigeria are most affected. An investigation was carried out to determine the quality of Pentazocine and Diazepam injections in circulation in Anambra State. The injections were sampled from different markets, shops, and pharmaceutical companies within the State. Imported and locally manufactured brands of the injections were analysed. Identification, pH, Assays, Bacteria Endotoxin, sterility, and sub-visible particulate matter contamination were analysed. The results of the Identification, Bacteria Endotoxin (BET), Particulate matter count, and pH are all within the British Pharmacopeia. The Pentazocine Assays obtained were (98.31%- 102.5%) which are within the British Pharmacopeia limit of (95% – 105%). This is good for health management. The Assays of two of the Diazepam injections tested were between (111.33% - 111.53%), which is above the approved British Pharmacopeia limit of (90%-110%). This is an indication of high level of impurities and not good for health care management.

A Mechanistic Study on the Fluidization and Enhancement Effects of Cineole Toward Stratum Corneum Intercellular Lamellar Lipids: A Liquid Crystalline Model Approach.

The stratum corneum (SC) serves as the primary barrier for permeation in human skin. Penetration enhancers, such as 1,8-cineole, are utilized to enhance the permeation of drugs. Cineole increases the permeation of chemicals through different mechanisms. However, its mechanism, particularly at high concentrations, has not been well-studied and is the subject of the present investigation. In continuation of our previous studies, the present investigation aims to elucidate the mechanism of action and concentration dependency of the effects of 1,8-cineole on the structure, diffusional properties, and partitioning behavior of the SC at high concentrations. This will be achieved through lamellar liquid crystalline models and ex-vivo skin studies. A lamellar liquid crystalline lipid matrix model in the presence (25 - 90%, w/w) and absence of cineole was prepared from SC lipids and characterized by X-ray diffraction, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and polarized light microscopy (PLM) studies. Release of the model lipophilic drug (diazepam) from cineole and cineole-treated matrices and the permeation of the drug from cineole and cineole-containing matrices (as a vehicle similar to the stratum corneum lipids) through excised rat skin were studied. Drug assay was performed by HPLC. The PLM, DSC, and X-ray studies showed that the model matrix had a lamellar gel-liquid crystalline structure, and cineole fluidized the structure concentration-dependently and created other mesomorphic textures, such as myelinic figures. Release experiments showed that diffusion coefficients remained almost constant at high cineole concentrations of 40-90%, suggesting similar fluidization states. Skin permeation studies indicated that the diffusion coefficient (estimated from lag-time) increased concentration-dependently and played a role in permeability coefficient (Kp) increments alongside the increased partitioning of the model drug into the skin. Data suggest that high concentrations of cineole at the skin surface might not provide enough cineole in the skin for full fluidization, despite the similarity of the vehicle to SC lipids and even at high concentrations. The enhancement effect of cineole is concentration-dependent and might reach maximum fluidization at certain concentrations, but this maximum might not be easily achievable when cineole is used in formulations as pure or in a vehicle.

A Mechanistic Study on the Fluidization and Enhancement Effects of Cineole Toward Stratum Corneum Intercellular Lamellar Lipids: A Liquid Crystalline Model Approach

Background: The stratum corneum (SC) serves as the primary barrier for permeation in human skin. Penetration enhancers, such as 1,8-cineole, are utilized to enhance the permeation of drugs. Cineole increases the permeation of chemicals through different mechanisms. However, its mechanism, particularly at high concentrations, has not been well-studied and is the subject of the present investigation. Objectives: In continuation of our previous studies, the present investigation aims to elucidate the mechanism of action and concentration dependency of the effects of 1,8-cineole on the structure, diffusional properties, and partitioning behavior of the SC at high concentrations. This will be achieved through lamellar liquid crystalline models and ex-vivo skin studies. Methods: A lamellar liquid crystalline lipid matrix model in the presence (25 - 90%, w/w) and absence of cineole was prepared from SC lipids and characterized by X-ray diffraction, differential scanning calorimetry (DSC), Thermogravimetric Analysis (TGA), and Polarized Light Microscopy (PLM) studies. The release of the model lipophilic drug (diazepam) from cineole and cineole-treated matrices and the permeation of the drug from cineole and cineole-containing matrices (as a vehicle similar to the stratum corneum lipids) through excised rat skin were studied. A drug assay was performed by HPLC. Results: The PLM, DSC, and X-ray studies showed that the model matrix had a lamellar gel-liquid crystalline structure, and cineole fluidized the structure concentration dependently and created other mesomorphic textures, such as myelinic figures. Release experiments showed that diffusion coefficients remained almost constant at high cineole concentrations of 40-90%, suggesting similar fluidization states. Skin permeation studies indicated that the diffusion coefficient (estimated from lag-time) increased concentration-dependently and played a role in permeability coefficient (Kp) increments alongside the increased partitioning of the model drug into the skin. Data suggest that high concentrations of cineole at the skin surface might not provide enough cineole in the skin for full fluidization, despite the similarity of the vehicle to SC lipids and even at high concentrations. Conclusions: The enhancement effect of cineole is concentration-dependent and might reach maximum fluidization at certain concentrations, but this maximum might not be easily achievable when cineole is used in formulations as pure or in a vehicle.

In Silico and In Vitro Screening of Novel Pyridazine Analogs as Muscle Relaxant Agent on Acetylcholine Muscarinic Receptor

Background:: Among Nitrogen-containing heterocyclic compounds, pyridazine derivatives serve as a necessary scaffold as they possess various pharmacological activities. Thus, in recent times, the design of novel synthetic schemes and the selection of a new target for the action of pyridazine derivatives have attracted the attention of researchers. Objective:: This study has focused on synthesizing and evaluating the muscle relaxant activity of pyridazine analogs by in-silico screening and rotarod test. Methods: In the present work, pyridazine derivatives were synthesized from substituted pyridine and maleic anhydride yielding intermediates (1a-5a), which on reaction with hydrazine, yielded final pyridazine derivatives (1b-5b). They were then screened for muscle relaxant action by an in-silico docking study against muscarinic acetylcholine receptors with protein data bank ID: 5CXV with the use of Autodock 4.2 and Biovia discovery studio tools. Compounds were further tested for muscle relaxant activity by the rotarod test. Results: Synthesis of the designed compounds was carried out successfully. Obtained result showed that the final compounds (1b-5b) showed 1-3 interactions with acetylcholine muscarinic receptor with -7.2 to -7.9 Kcal/mole affinities. The findings were compared to the typical drug diazepam, which has one interaction with the target and binding energy of -7.7 Kcal/mole. Moreover, the result of the rotarod test showed that substitution by electron-withdrawing groups causes more muscle relaxant activity when compared with the electron releasing groups. Conclusion:: The results of the experimental study showed that pyridazine derivatives could serve as a promising template for the further design and development of muscle relaxant agents.

Intralipid as a matrix additive for evaluating hyperlipidemic postmortem blood.

Postmortem whole blood samples can differ greatly in quality where hyperlipemia is a frequent variable that can influence the results of analytical methods. The aim of this study was to investigate the influence of lipemia on postmortem analysis as well as demonstrate the usage of Intralipid in comparison to pooled postmortem lipids as matrix additives for meaningful evaluation and validation of hyperlipidemic postmortem samples. Hyperlipidemic blood samples were simulated by adding different concentrations of Intralipid or pooled authentic postmortem lipids to bovine whole blood. The hyperlipidemic blood samples were spiked with 14 benzodiazepines and five sedative and antianxiety drugs (alprazolam, clonazepam, 7-aminoclonazepam, diazepam, flunitrazepam, 7-aminoflunitrazepam, hydroxyzine, lorazepam, midazolam, nitrazepam, 7-aminonitrazepam, nordazepam, oxazepam, propiomazine, dihydropropiomazine, temazepam, triazolam, zolpidem and zopiclone). Samples were prepared with liquid-liquid extraction followed by ultra-high performance liquid chromatography-mass spectrometry. The effects of lipemia on the recovery of analytes and internal standards (ISs) were evaluated to determine the effect of, and any differences between, the two additives. Lipemia was found to cause major interference when quantifying the analytes. For most analytes, the ISs could compensate for analyte losses. However, the most hydrophilic analytes (7-amino metabolites), together with the most lipophilic analytes (propiomazine and dihydropropiomazine), were greatly affected by lipemia (<50% recovery), and the IS could not compensate for analyte losses. In general, lower analyte recoveries were observed for samples with Intralipid as a lipemic additive in comparison to those containing pooled postmortem lipids. Both Intralipid and pooled postmortem lipids showed marked effects on the analytical results. Intralipid gave a good indication of the effects of lipemia and could be a useful tool for making a meaningful evaluation of hyperlipidemic postmortem samples during the method development and validation.