Glutamate Carboxypeptidase II in Aging Rat Prefrontal Cortex Impairs Working Memory Performance

Glutamate carboxypeptidase II (GCPII) expression in brain is increased by inflammation, and reduces NAAG (N-acetyl aspartyl glutamate) stimulation of mGluR3 signaling. Genetic insults in this signaling cascade are increasingly linked to cognitive disorders in humans, where increased GCPII and or decreased NAAG-mGluR3 are associated with impaired prefrontal cortical (PFC) activation and cognitive impairment. As aging is associated with increased inflammation and PFC cognitive deficits, the current study examined GCPII and mGluR3 expression in the aging rat medial PFC, and tested whether GCPII inhibition with 2-(3-mercaptopropyl) pentanedioic acid (2-MPPA) would improve working memory performance. We found that GCPII protein was expressed on astrocytes and some microglia as expected from previous studies, but was also prominently expressed on neurons, and showed increased levels with advancing age. Systemic administration of the GCPII inhibitor, 2-MPPA, improved working memory performance in young and aged rats, and also improved performance after local infusion into the medial PFC. As GCPII inhibitors are well-tolerated, they may provide an important new direction for treatment of cognitive disorders associated with aging and/or inflammation.

Synthesis and application of stationary phase for DNA-affinity chromatographic analysis of unmodified and antisense oligonucleotide

The goal of the research was the synthesis and application of an oligonucleotide immobilized stationary phase for the analysis of unmodified and antisense oligonucleotides. The method for attaching these molecules to aminopropyl silica modified with pentanedioic acid was developed. Each step of the synthesis was carefully controlled with the application of spectroscopic, elemental, and chromatographic analyses. The oligonucleotide-based stationary phase was applied for the retention studies. Unmodified oligonucleotides of different complementarity to the molecule attached as a stationary phase, as well as antisense oligonucleotides, were tested. The comparative study upon complex optimization of oligonucleotide analysis in different liquid chromatography modes was performed. Results have shown that this stationary phase may be applied for oligonucleotide analysis in hydrophilic interaction liquid chromatography and ion exchange chromatography, but no unique sequence-based selectivity was obtained. Contrary results were observed for affinity chromatography, which allowed for specific separation of the complementary strands based on hydrogen bonding and stacking interactions, where the temperature was the main factor influencing the selectivity of the separation. Furthermore, the oligonucleotide-based stationary phase may be applied for comparative antisense oligonucleotide hybridization studies to a specific RNA sequence. All of the results have shown that affinity chromatography with oligonucleotide-based stationary phases is a powerful technique for the specific base recognition of polynucleotides. Graphical abstract

Pharmacokinetics of Pentanedioic Acid Imidazolyl Ethanamide in Healthy Volunteers

Relevance. Pentanedioic acid imidazolyl ethanamide (PAIE) has been used clinically as an antiviral agent for a long time, however, there is no information in the available literature concerning the dependence of PAIE pharmacokinetics on isoenzymes polymorphism of the cytochrome P450 system (CYP), as well as on the variability of pharmacokinetic parameters in humans. The aim of the study is to determine the main pharmacokinetic parameters of PAIE in healthy volunteers and to assess the contribution of polymorphism of CYP P450 isoenzymes to the variability of pharmacokinetic parameters.Material and methods. The study included 12 healthy volunteers (5 men and 7 women) of the Caucasian race, who took In[1]gavirin® at a dose of 180 mg (2 capsules of 90 mg) on an empty stomach during two dosing periods, separated by a 7-day washout period. Determination of PAIE concentration in blood plasma and urine samples was carried out by high-performance liquid chromatography–mass spectrometry. Polymorphism of CYP genes was analyzed using the polymerase chain re[1]action method in order to analyze the pharmacogenetic features of PAIE metabolism in volunteers during the study.Results. After oral administration, PAIE quickly reached the systemic circulation: the maximum concentration of 578.88±145.21 ng/ml was observed after about 2 hours. Pharmacokinetic parameters of PAIE did not show high intraindividual variability and did not depend on polymorphism of isoenzymes CYP1A1, CYP2C9, and CYP2D6. Within 48 hours after the administration of the studied drug, about half of the taken dose of PAIE was excreted in the urine unchanged, which indicates a significant contribution of the kidneys to the elimination of PAIE. The only adverse event registered in 1 volunteer was a clinically insignificant decrease in the level of leukocytes, which did not require medical intervention and was resolved without consequences.Conclusion. PAIE is characterized by predictable pharmacokinetics, low intraindividual variability of pharmacokinetic parameters, and a favorable safety profile.

Clinical Efficacy and Safety of Pentanedioic Acid Imidazolyl Ethanamide in Patients Aged 3 to 6 years with Influenza and other Acute Respiratory Viral Infections Based on the Results of Double-Blind Randomized Placebo-Controlled Multicenter Study

Study Goals. Assessment of the efficacy and safety of the Pentanedioic Acid Imidazolyl Ethanamide (IPA) in the treatment of influenza and other acute respiratory viral infections (ARVI) in children aged 3–6 years. Children Characteristics and Study Methods. The randomized double-blind placebo-controlled multicenter clinical study involving 190 patients aged 3–6 years was carried out as follows: 95 patients received IPA at 30 mg/day dose once for 5 days and 95 patients – placebo using the same pattern. The therapy efficacy was assessed by the body temperature, dynamics of the individual symptoms of the disease as per the Severity Scale for the patients with influenza and other acute respiratory viral infections (Scale) and the incidence rate of complications of the acute respiratory viral infections. The primary endpoint is the period of the score reduction on the Scale to 2 points, providing that there is not more than 1 score on the individual subscales with the body temperature normalization from the start of treatment. The safety analysis was carried out using the assessment of the nature and incidence rate of the adverse events. Results. The IPA use at 30 mg/day dose significantly promotes the achievement of goals for the primary endpoint – the average value in the IPA group is 91.79 h (95% confidence interval – CI from 87.45 to 96.13), in the placebo group – 100.12 h (95% CI from 96.73 to 103.51). In the IPA group, the body temperature returned to normal by 18.56 h faster in average compared with the placebo. The regression of the catarrhal and intoxication symptoms was significantly promoted with the IPA use as follows: by the 3rd day of the therapy, the average score on the Scale was 5.22 points for IPA, and it was statistically significantly lower than that in the placebo group – 6.21 points. The comparative analysis of the incidence rate of adverse events did not reveal the statistically significant differences between IPA and placebo. None of the adverse events recorded was clinically significant, and none caused the cancellation or change in the dosage of the study drug. Conclusions. IPA broad-spectrum antiviral drug at 30 mg/day dose has demonstrated the high efficacy in the treatment of the acute respiratory viral infections of various etiologies in children aged 3–6 years. The IPA use compared with the placebo causes the significant reduction of the fever period, accelerates the relief of the intoxication and catarrhal symptoms, and decreases the recovery time. The IPA is characterized by high tolerability and high safety, which allows to recommend the drug for the treatment of influenza and ARVI (acute respiratory viral infections) in children from 3 years of age.

Design, Synthesis and in vitro Evaluation of 2-(Quinoline-8-sulfonamido)pentanedioic Acid Analogues as Antiangiogenic and Antitumor Agents on Multiple Myeloma

Thalidomide is presently approved as antiangiogenic and anticancer drug in multiple myeloma. The authors present a number of analogue-based designs of N-(o-carboxybenzoyl)-DL-glutamic acid, a major thalidomide metabolite. The compounds were synthesized and biologically tested in multiple myeloma as anticancer agents. Three compounds inhibited HUVEC proliferation at low micromolar concentrations, indicating that they are antiangiogenic and cytotoxic to human multiple myeloma RPMI8226. The active compounds were tested for antiproliferative activity on HUVECs using the dye exclusion method with trypan blue. Dimethyl-2-(quinoline-8-sulfonamido)pentanedioate (2c), in particular, inhibits VEGFR-2 phosphorylation at the Tyr-1175 residue, as determined by SDS PAGE. The binding mode of (2c) was predicted in silico in order to better understand how it interacts with essential amino acid residues in the VEGFR-2 active site. The binding energy was calculated as -161.41kcal/mol. in vitro Study of the compounds on the Vero cell line shows less toxicity towards the normal endothelial cells than the cancer cells.