PHARMACOKINETIC DRUG INTERACTIONS OF GLICLAZIDE AND ITOPRIDE IN NORMAL AND DIABETIC RATS

Objective: The present study was aimed to investigate the safety, reliability of Gliclazide and possible drug interaction with Itopride when they were administered as combination treatment.Methods: Studies were conducted in normal and streptozotocin induced diabetic rats with oral administration of selected doses of gliclazide, itopride and their combination. Blood samples were collected from rats by retro orbital/marginal ear vein puncture at regular intervals of time. All the blood samples were analyzed for pharmacokinetic parameters by HPLC method.Results: There was no significant difference in pharmacokinetic parameters of both Gliclazide alone and combination with itopride in healthy and diabetic rats on day 1 and day 8.Conclusion: Based on the results it can be concluded that the concurrent administration of these two drugs have potential benefit without any drug interactions in the effective management of diabetes and gastroparesis.Â

Changes in liver lipids after administration of 2‐decanoylamino‐3‐morpholinopropiophenone and chlorpromazine

The enzyme which forms glucocerebroside, ceramide: UDP‐glucose glucosyltransferase, is inactivated in vitro by a cationic analog of cerebroside, 2‐decanoylamino‐3‐morpholinopropiophenone. A study of the inhibitor using intraperitoneal injection into young mice showed that the level of the enzyme activity in liver was appreciably lowered between 3 and 6 hr after injection. The activity increased subsequently, overshooting the normal level within 24 hr by about 20%, then returning to normal within the next 24 hr. Additional effects observed in liver were an increase in lipid content (primarily in the triglyceride fraction and ceramides) and a decrease in the glucocerebroside level. Body temperature dropped rapidly. Markedly similar effects were produced by injecting chlorpromazine, which was tried in order to reduce the hyperirritability and inhibitory effects on monoamine oxidase previously demonstrated by the glucosyltransferase inhibitor. Chlorpromazine did indeed block the hyperirritability and resulted in enhancement of the keto amine’s effects on the enzyme and lipids. It is possible that the two drugs in combination would be helpful in ameliorating the symptoms due to the cerebroside accumulation that occurs in Gaucher disease. Diazepam also produced a reduced level of glucosyltransferase. A color reaction for chlorpromazine, possibly suitable for quantitative determination in tissues, was accidentally discovered.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/142035/1/lipd0538.pd

Analogs of ceramide that inhibit glucocerebroside synthetase in mouse brain

In a search for potent inhibitors of glucocerebroside biosynthesis, we synthesized aromatic analogs of the enzyme's substrate, ceramide, many of which have not previously been described in the literature. Mouse brain and spleen, rat brain, and human placenta and spleen were all found to be susceptible to inhibition by a variety of compounds, although to differing extents. The most potent inhibitor was 2-decanoylamino-3-morpholino-1-phenylpropanol. The dehydro version of this compound (2-decanoylamino-3-morpholinopropiophenone) was less effective but it produced inactivation of the enzyme, probably by covalent reaction with the enzyme's active site. Examination of the various effects seen leads us to suggest that the active region of the enzyme contains four recognitional sites: an anionic moiety that may bind the glucose in activated form, an oxygen-binding region oriented toward the third carbon atom of ceramide, a narrow region that binds the alkyl chain of the fatty acid moiety, and a less narrow region that binds the hydrocarbon chain of the sphingoid base moiety.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23277/1/0000214.pd

A New Class of Monoamine Oxidase Inhibitors

Newly synthesized compounds have been found to inhibit mitochondrial monoamine oxidase (MAO) in mouse brain and rat liver. A series of 2-acylamino-3- tert -aminopropiophenones acted preferentially against MAO type B (2-phenylethylamine as substrate), apparently irreversibly. 2-Decanoylamino-3-morpholinopropiophenone acted similarly in vivo toward the cerebral MAO, producing a dose-related inhibition. At high dose levels, MAO type A was also severely inhibited. The effects were produced rapidly and restoration of enzyme activity also appeared rapidly. The half-life for MAO type A could be estimated from the rate of enzyme reappearance to be 13 h. It is suggested that the amino ketones undergo a Β-elimination reaction at the enzyme's active site, forming a reactive species (an Α,Β-unsaturated ketone), which reacts covalently with a nucleophilic group of the enzyme by a Michael addition. Some other related compounds, derivatives of phenylpropane, also showed inhibitory activity against MAO, particularly against type A (serotonin as substrate). The morpholino compound might have promise as a quickly effective, short-acting inhibitor of MAO type B.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66333/1/j.1471-4159.1980.tb06611.x.pd

Pharmacokinetic interactions between concomitantly administered Metformin and Itopride in rats

Gastroparesis is a syndrome characterized by delayed gastric emptying, in the absence of mechanical obstruction of the stomach. Diabetes mellitus (DM) is responsible for almost one third of cases of gastroparesis. Gastroparesis typically develops after at least 10 years of evolution of DM, and often is associated with other complications. The present study aimed to investigate the safety, reliability of Metformin and possible drug interaction with Itopride when they were administered as combination treatment. The study was conducted on healthy Wistar and streptozotocin induced diabetic rats.  A simple and sensitive high performance liquid chromatographic method was developed for the simultaneous estimation of Metformin and Itopride in rat plasma and also to estimate possible pharmacokinetic parameters of these drugs after oral administration. There was no significant difference in the Metformin alone and combination with Itopride and Itopride alone and combination with Metformin on day 1 and day 8 respectively. There is no significant change in tmax, cmax, AUC (0 – t) and AUC (o-inf), t½, Cl/f and V/f on day 1 and day 8 respectively in both diabetic and healthy rats. From the above results it can be concluded that the concurrent administration of these two drugs have potential benefit in the treatment of Diabetes and Gastroparesis. In addition, due to their insignificant pharmacokinetic interaction the combinational therapy can be safe and highly advantageous in Gastro paresis patients with diabetes. 

Phenotypic changes in 3T3 cells associated with the change of sphingolipid synthesis by a ceramide analog, 2-decanoylamino-3-morpholino-1-phenylpropanol (compound RV538)

A culture of BALB/c 3T3 cells grown in the presence of 40 [mu]M of the ceramide analog compound RV538 (2-decanoylamino-3-morpholino-1-phenylpropanol) for several passages caused a substantial decrease in the level of all glycosphingolipids and an accumulation of ceramide and sphingomyelin. Associated with these chemical changes of sphingolipid composition and metabolism, the following phenotypic changes were observed: (i) loss of the cobblestone appearance at cell density saturation and development of fibroblastic appearance with partial overlapping of cells; (ii) reduction of cell growth rate; (iii) enhanced production of lactic acid; (iv) enhanced rate of glucose transport; and (v) higher incidence of large colony formation with infiltrating appearance in soft agar. Cell morphology changes, lactate pruduction, and enhanced sugar uptake were reversed by co-culturing cells with gangliosides, particularly trisialogangliosides. Thus, these phenotypic changes mimicking those of oncogenically transformed cells are closely related to the blocked synthesis of glycolipids in these cells, whereas other changes may be caused by an accumulation of ceramide and sphingomyelin.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27183/1/0000186.pd

Antimicrobial activity of nature-inspired molecules against multidrug-resistant bacteria

Multidrug-resistant bacterial infections present a serious challenge to global health. In addition to the spread of antibiotic resistance, some bacteria can form persister cells which are tolerant to most antibiotics and can lead to treatment failure or relapse. In the present work, we report the discovery of a new class of small molecules with potent antimicrobial activity against Gram-positive bacteria and moderate activity against Gram-negative drug-resistant bacterial pathogens. The lead compound SIMR 2404 had a minimal inhibitory concentration (MIC) of 2 μg/mL against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-intermediate Staphylococcus aureus (VISA). The MIC values against Gram-negative bacteria such as Escherichia coli and Actinobacteria baumannii were between 8–32 μg/mL. Time-kill experiments show that compound SIMR 2404 can rapidly kill tested bacteria. Compound SIMR 2404 was also found to rapidly kill MRSA persisters which display high levels of tolerance to conventional antibiotics. In antibiotic evolution experiments, MRSA quickly developed resistance to ciprofloxacin but failed to develop resistance to compound SIMR 2404 even after 24 serial passages. Compound SIMR 2404 was not toxic to normal human fibroblast at a concentration of 4 μg/mL which is twice the MIC concentration against MRSA. However, at a concentration of 8 μg/mL or higher, it showed cytotoxic activity indicating that it is not ideal as a candidate against Gram-negative bacteria. The acceptable toxicity profile and rapid antibacterial activity against MRSA highlight the potential of these molecules for further studies as anti-MRSA agents

All-d-Enantiomer of β-Amyloid Peptide Forms Ion Channels in Lipid Bilayers

Alzheimer’s disease (AD) is the most common type of senile dementia in aging populations. Amyloid β (Aβ)-mediated dysregulation of ionic homeostasis is the prevailing underlying mechanism leading to synaptic degeneration and neuronal death. Aβ-dependent ionic dysregulation most likely occurs either directly via unregulated ionic transport through the membrane or indirectly via Aβ binding to cell membrane receptors and subsequent opening of existing ion channels or transporters. Receptor binding is expected to involve a high degree of stereospecificity. Here, we investigated whether an Aβ peptide enantiomer, whose entire sequence consists of d-amino acids, can form ion-conducting channels; these channels can directly mediate Aβ effects even in the absence of receptor–peptide interactions. Using complementary approaches of planar lipid bilayer (PLB) electrophysiological recordings and molecular dynamics (MD) simulations, we show that the d-Aβ isomer exhibits ion conductance behavior in the bilayer indistinguishable from that described earlier for the l-Aβ isomer. The d isomer forms channel-like pores with heterogeneous ionic conductance similar to the l-Aβ isomer channels, and the d-isomer channel conductance is blocked by Zn2+, a known blocker of l-Aβ isomer channels. MD simulations further verify formation of β-barrel-like Aβ channels with d- and l-isomers, illustrating that both d- and l-Aβ barrels can conduct cations. The calculated values of the single-channel conductance are approximately in the range of the experimental values. These findings are in agreement with amyloids forming Ca2+ leaking, unregulated channels in AD, and suggest that Aβ toxicity is mediated through a receptor-independent, nonstereoselective mechanism

Short chain ceramides as substrates for glucocerebroside synthetase. Differences between liver and brain enzymes

In order to increase the sensitivity of the assay for ceramide: UDPGlc glucosyltransferase, the enzyme that makes glucocerebroside, we synthesized a variety of ceramide homologues that might be better substrates than the naturally occurring ceramides. N-Octanoyl sphingosine proved to be the best lipid tested in liver and brain. It could be added to the tissue homogenate in the dry form, as a thin layer coated on Celite, or in liposomes, prepared from lecithin and cerebroside sulfate. The liposomal form produced better replication of assay values. It is suggested that the addition of cerebroside sulfate to liposomal preparations might be a good, and more physiological, replacement for the commonly used dicetyl phosphate.A new homologue of DL-sphinganine, decasphinganine, was synthesized by an efficient series of steps and acylated with different fatty acids to form ceramide homologues. The best substrate in this series was the lauroyl amide and it is suggested that this lipid be used in cerebroside synthetase assays because of the convenience of preparing it, even though it is not as good as octanoyl sphingosine. Both compounds are distinctly better than natural ceramide or DL-sphinganine amides.From comparisons of enzyme activity under various conditions, the tentative conclusion is drawn that the enzymes in liver and brain have different properties, and that liver has two different synthetases.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23580/1/0000542.pd