Ryan da Silva Ramos

Identification of Novel Protein Kinase Receptor Type 2 Inhibitors Using Pharmacophore and Structure-Based Virtual Screening

The Protein Kinase Receptor type 2 (RIPK2) plays an important role in the pathogenesis of inflammatory diseases; it signals downstream of the NOD1 and NOD2 intracellular sensors and promotes a productive inflammatory response. However, excessive NOD2 signaling has been associated with various diseases, including sarcoidosis and inflammatory arthritis; the pharmacological inhibition of RIPK2 is an affinity strategy that demonstrates an increased expression of pro-inflammatory secretion activity. In this study, a pharmacophoric model based on the crystallographic pose of ponatinib, a potent RIPK2 inhibitor, and 30 other ones selected from the BindingDB repository database, was built. Compounds were selected based on the available ZINC compounds database and in silico predictions of their pharmacokinetic, toxicity and potential biological activity. Molecular docking was performed to identify the probable interactions of the compounds as well as their binding affinity with RIPK2. The compounds were analyzed to ponatinib and WEHI-345, which also used as a control. At least one of the compounds exhibited suitable pharmacokinetic properties, low toxicity and an interesting binding affinity and high fitness compared with the crystallographic pose of WEHI-345 in complex with RIPK2. This compound also possessed suitable synthetic accessibility, rendering it a potential and very promising RIPK2 inhibitor to be further investigated in regards to different diseases, particularly inflammatory ones.

Virtual Screening and Statistical Analysis in the Design of New Caffeine Analogues Molecules with Potential Epithelial Anticancer Activity

About 132 thousand cases of melanoma (more severe type of skin cancer) were registered in 2014 according to the World Health Organization. This type of cancer significantly affects the quality of life of individuals. Caffeine has shown potential inhibitory effect against epithelial cancer. In this study, it was proposed to obtain new caffeine-based molecules with potential epithelial anticancer activity. For this, a training set of 21 molecules was used for pharmacophore perception procedures. Multiple linear regression analyses were used to propose mono-, bi-, tri-, and tetra-parametric models applied in the prediction of the activity. The generated pharmacophore was used to select 350 molecules available at the ZINCpharmer server, followed by reduction to 24 molecules, after selection using the Tanimoto index, yielding 10 molecules after final selection by predicted activity values > 1.5229. These ten mole-cules had better pharmacokinetic properties than the other ones used as reference and within the clinical-ly significant limits. Only two molecules show minor hits of toxicity and were submitted to molecular docking procedures, showing BFE (binding free energy) values lower than the reference values. Statisti-cal analyses indicated strong negative correlations between BFE and pharmacophoric properties (high influence on BFE lowering) and practically null correlation between BFE and BBB. The two most prom-ising molecules can be indicated as candidates for further in vitro and in vivo analyzes.

Oil from the fruits of Pterodon emarginatus Vog.: A traditional anti-inflammatory. Study combining in vivo and in silico

ETHNOBOTANICAL RELEVANCE The oil obtained from the fruits of Pterodon emarginatus Vog. (OPe) is used orally and topically, in traditional medicine for some purposes, such as acute and chronic inflammatory states as rheumatoid arthritis. MATERIALS AND METHODS In this work, the anti-inflammatory activity of the OPe was demonstrated based on several animal models and presented an in silico study based on the 6α,7β-dihydroxy-vouacapan-17β-oic acid (DHVA) majority compound of the OPe to evaluate the interaction this compound, with cyclooxygenase-2 (COX-2) in 4COX (Mus musculus) and 5KIR (Homo sapiens) and molecular dynamics simulation. RESULTS The OPe (498 mg/kg, p.o) significantly inhibited (p < 0.05, Student t-test) the primary and secondary reactions of arthritis by Freunds Complete Adjuvant (FCA) and in dermatitis induced by croton oil in mice, OPe inhibited peak of edema. In vascular permeability test in rats, the treatment with OPe was able to block the response to PGE2, serotonin, and bradykinin (p < 0.05, Student t-test). In the writhing test in mice, the OPe at doses of 498 and 980 mg/kg (p.o) produced inhibition of 73% and 92%, respectively, and was not significantly effective in the hot plate test. In the evaluation of the potency in relation to gastric injury (gastric ulcer induced by stress) and combined assay in the assessment of anti-inflammatory potency and gastric damage, it was observed that indomethacin (10 mg/kg, p.o.) inhibited carrageenan edema by 51% and produced a higher number of gastric lesions when compared to the group treated with OPe, where only areas of hyperemia were observed, without the occurrence of ulcerative lesion, and which inhibited the edema by 47%. In the in silico study, it was found that the DHVA is capable of binding to two organisms (4COX - Mus musculus and 5KIR - Homo sapiens), however, with higher binding affinity to the organism Homo sapiens. CONCLUSIONS As expected, all tested ligands were capable of forming hydrogen interactions with residues at their respective binding sites, but the DHVA ligand was capable of creating slightly more hydrogen bonds when docked to either 4COX or 5KIR than the other tested ligands, thus demonstrating the participation of this compound in the anti-inflammatory and antialgic responses observed in the in vivo assays as a COX-2 inhibitor. Therefore, the results obtained support the traditional use of OPe for inflammatory and gastric problems.

An Antioxidant Potential, Quantum-Chemical and Molecular Docking Study of the Major Chemical Constituents Present in the Leaves of Curatella americana Linn

Reactive oxygen species (ROS) are continuously generated in the normal biological systems, primarily by enzymes as xanthine oxidase (XO). The inappropriate scavenging or inhibition of ROS has been considered to be linked with aging, inflammatory disorders, and chronic diseases. Therefore, many plants and their products have been investigated as natural antioxidants for their potential use in preventive medicine. The leaves and bark extracts of Curatella americana Linn. were described in scientific research as anti-inflammatory, vasodilator, anti-ulcerogenic, and hypolipidemic effects. So, the aim of this study was to evaluate the antioxidant potentials of leaf hydroalcoholic extract from C. americana (HECA) through the scavenging DPPH assay and their main chemical constituents, evaluated by the following quantum chemical approaches (DFT B3LYP/6-31G**): Maps of Molecular Electrostatic Potential (MEP), Frontier Orbital’s (HOMO and LUMO) followed by multivariate analysis and molecular docking simulations with the xanthine oxidase enzyme. The hydroalcoholic extract showed significant antioxidant activity by free radical scavenging probably due to the great presence of flavonoids, which were grouped in the PCA and HCA analysis with the standard gallic acid. In the molecular docking study, the compounds studied presented the binding free energy (ΔG) values close each other, due to the similar interactions with amino acids residues at the activity site. The descriptors Gap and softness were important to characterize the molecules with antioxidant potential by capturing oxygen radicals.

VALIDATION OF COMPUTATIONAL METHODS APPLIED IN MOLECULAR MODELING OF CAFFEINE WITH EPITHELIAL ANTICANCER ACTIVITY: THEORETICAL STUDY OF GEOMETRIC, THERMOCHEMICAL AND SPECTROMETRIC DATA

Josivan da Silva Costaa,b,c,*, Cleydson Breno Rodrigues dos Santosa,b, Karina da Silva Lopes Costab, Ryan da Silva Ramosb, Carlos Henrique Tomich de Paula da Silva,d and Williams Jorge da Cruz Macêdoc Universidade Federal do Pará, Rua Augusto Corrêa, 1 Guamá, 66075-110 Belém – PA, Brasil Departamento de Ciências Biológicas, Universidade Federal do Amapá, Rod. Juscelino Kubitschek, Km 02, s/n, Jardim Marco Zero, 68902-280 Macapá – AP, Brasil Universidade Federal Rural da Amazônia, Rua João Pessoa, 121, Campus Capanema Centro, 68700-030 Capanema – PA, Brasil Escola de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, 14040-903 Ribeirão Preto – SP, Brasil

BLOOD PRESSURE (BP) OUTCOME IN PATIENTS WITH SECONDARY ANEMIA IN PERITONEAL DIALYSIS TREATED WITH METOXIPOLIETILENGLICOL-EPOETINA (MIRCERA®) (CAPRI STUDY): PP.9.379

Objectives: To study the outcome of BP in patients with secondary anemia related to End Stage Renal Disease (ESRD) in peritoneal dialysis (PD), treated with MIRCERA. Methods: Observational and prospective study performed in PD patients of several hospitals in Catalonian. We included 49 patients naive or treated previously with darboepoetina or Beta epoetina. They started MIRCERA after beginning PD, and we performed a follow-up of demographical and analytical data and BP for a 6 months. Results: Between the 49 patients, 17 are male (34.7%). Age = 58.5 ± 15.6 years. The etiology of ESRD was glomerulopathy 32,7%, in 28.6% Diabetes Mellitus,16.3% unknown, 10.2% polychystosis and other etiology: 12,2%. 14 have cardiovascular disease, 20 hypercholesterolemia,1 cerebrovascular disease and 2 oncological events. 65.3% patients start PD as first treatment, 18.4 % come from hemodialysis and 16.3% from kidney transplant failure. 93.9% have high BP, 85.7% on pharmacological treatment. 14 were treated with ACEI, 4 with ARAII and 20 with diuretics. The starting dose of Mircera was 50 μg once monthly in 4.1%, 75 in 26.5%, 100 μg in 24.5%, 150 μg in 20.4%, 200 in 10.2% and 250 in 8.2% of cases. We observe that 29.8% have Hb levels <11 g/dl, 51.1% between 11–12.99 g/dl and 19.1% ± 13 g/dl at the beginning of the study. At 6 months the distribution was 19.1%, 53.2% and 27.7% respectively and no significance differences were found. There were no differences in Mircera doses between the patients treated with ACEI or ARA II and they treated with other drugs. If we consider hypertension as SBP ± 140 and DBP ± 90 mmHg, at the beginning 63.3% were hypertensive and 49.0% (p = 0.046)at the end. Only 2 patients with initial normal BP (140/90) have an increase at 6months. 9 patients with initial hypertension decrease BP levels to normal range; (p global = 0.065, p per groups of doses = 0.859)).Table 2 Figure 1. No caption available. Figure 2. No caption available. Conclusions: Treatment with MIRCERA in PD patients allows a good control of anaemia without negative influence in the control of BP.