Synthesis, biological activity, and ADME properties of novel S-DABOs/N-DABOs as HIV reverse transcriptase inhibitors

Previous studies aimed at exploring the SAR of C2-functionalized S-DABOs demonstrated that the substituent at this position plays a key role in the inhibition of both wild-type RT and drug-resistant enzymes, particularly the K103N mutant form. The introduction of a cyclopropyl group led us to the discovery of a potent inhibitor with picomolar activity against wild-type RT and nanomolar activity against many key mutant forms such as K103N. Despite its excellent antiviral profile, this compound suffers from a suboptimal ADME profile typical of many S-DABO analogues, but it could, however, represent a promising candidate as an anti-HIV microbicide. In the present work, a new series of S-DABO/N-DABO derivatives were synthesized to obtain additional SAR information on the C2-position and in particular to improve ADME properties while maintaining a good activity profile against HIV-1 RT. In vitro ADME properties (PAMPA permeation, water solubility, and metabolic stability) were also experimentally evaluated for the most interesting compounds to obtain a reliable indication of their plasma levels after oral administration.     

Capillary extrusion behavior of phenolphthalein poly(ether-ether-sulphone) (PES-C)

The capillary extrusion flow properties of novel engineering thermoplastic phenolphthalein poly(ether-ether-sulphone) (PES-C) have been investigated using capillary rheometer. The dependence of viscosity on the wall shear rate and temperatures were obtained. The flow activation energy was found to decrease with shear rate but to be constant with shear stress. The entrance effect was calculated and from which the extensional behavior was estimated using Cogswell's method. From the extrudate swell ratio the principal normal stress was evaluated and a temperature-independent correlation was observed when they were plotted against shear stress. The melt fracture phenomena were checked and discussed also. © 1997 John Wiley & Sons, Inc. J Appl Polym Sci 65:951–958, 1997     

Effects of high glucose on vascular endothelial growth factor synthesis and secretion in aortic vascular smooth muscle cells from obese and lean zucker rats

Type 1 diabetes is characterized by insulin deficiency, type 2 by both insulin deficiency and insulin resistance: in both conditions, hyperglycaemia is accompanied by an increased cardiovascular risk, due to increased atherosclerotic plaque formation/instabilization and impaired collateral vessel formation. An important factor in these phenomena is the Vascular Endothelial Growth Factor (VEGF), a molecule produced also by Vascular Smooth Muscle Cells (VSMC). We aimed at evaluating the role of high glucose on VEGF-A(164) synthesis and secretion in VSMC from lean insulin-sensitive and obese insulin-resistant Zucker rats (LZR and OZR). In cultured aortic VSMC from LZR and OZR incubated for 24 h with d-glucose (5.5, 15 and 25 mM) or with the osmotic controls l-glucose and mannitol, we measured VEGF-A(164) synthesis (western, blotting) and secretion (western blotting and ELISA). We observed that: (i) d-glucose dose-dependently increases VEGF-A(164) synthesis and secretion in VSMC from LZR and OZR (n = 6, ANOVA p = 0.002-0.0001); (ii) all the effects of 15 and 25 mM d-glucose are attenuated in VSMC from OZR vs. LZR (p = 0.0001); (iii) l-glucose and mannitol reproduce the VEGF-A(164) modulation induced by d-glucose in VSMC from both LZR and OZR. Thus, glucose increases via an osmotic mechanism VEGF synthesis and secretion in VSMC, an effect attenuated in the presence of insulin resistance.     

Effect of polypropylene morphology on thermal cis-trans isomerization and photofading of some azo dyes: A kinetic study

Five polypropylene films were prepared having different crystallinity and morphology, the latter having been modified by stretching. They were colored with azo dyes XC₆H₄N?NC₆H₄N(C₂H₅)₂ (where X?H, OCH₃, CN, and NO₂). The kinetics of the thermal cis-trans isomerization of these dyes has been studied in the range 21–41°C, much above the glass transition temperature of the polymer. The isomerization process was found to be strictly first-order; the kinetic parameter values have been correlated with the free volume extent in the amorphous regions of the matrix. Lightfastness of the dyes in the polymer matrices has been also investigated: It appeared to be more important for the unoriented samples with respect to the stretched ones and substantially independent on the crystalinity degree.     

Rationally designed inhibitors as tools for comparing the mechanism of squalene-hopene cyclase with oxidosqualene cyclase

The inhibition of squalene-hopene cyclase (SHC) (E.C. 5.4.99.-), an enzyme of bacterial membranes catalyzing the formation of pentacyclic sterol-like triterpenes, was studied by using different classes of compounds originally developed as inhibitors of oxidosqualene cyclase (OCS) (E.C. 5.4.99.7), the enzyme of eukaryotes responsible for the formation of tetracyclic precursors of sterols. The mechanism of cyclization of squalene by SHC, beginning with a protonation of the 2,3 double bond by an acidic residue of the enzyme, followed by a series of electrophilic additions of the carbocationic intermediates to the double bonds, is similar to the mechanism of cyclization of 2,3-oxidosqualene by OSC. The inhibitors studied included: (i) analogs of the carbocationic intermediates formed during cyclization, such as aza-analogs of squalene and 2,3-oxidosqualene; (ii) affinity-labeling inhibitors bearing a methylidene reactive group; and (iii) vinyldioxidosqualenes and vinylsulfide derivatives of the substrates. Comparison of the results obtained with the two enzymes, SHC and OSC, showed that many of the most effective inhibitors of OSC were also able to inhibit SHC, while some derivatives acted as specific inhibitors. Differences could be easily explained on the basis of the different substrate specificity of the two enzymes.     

Advanced testing procedures for high performance coatings

In the first part of this work, an integrated approach incorporating electrochemical (electrochemical impedance spectroscopy), calorimetric (differential scanning calorimetry), mechanical (dynamic mechanical thermal analysis), and infrared spectroscopy (Fourier transform infrared spectroscopy) techniques was used to characterize some organic coatings from the automotive industry. Comparison of the results obtained with different techniques allows not only a higher precision in the evaluation of natural or artificial degradation, which is very difficult to obtain using traditional techniques, but also better data interpretation and, therefore, the possibility of understanding the different degradation mechanisms. In the second part, some preliminary results concerning the comparison of salt spray exposure and EIS measurements are discussed. The statistical analysis of the results showed that the degradation mechanism in the two cases is completely different and, therefore, the information which is obtained with the two testing approaches is also different. With EIS measurements obtained in immersion, it is possible to monitor, in particular, the barrier properties of the coatings, which are dominated by the presence of defects. In the case of salt spray (without scratch), the main property to be measured is the loss of adhesion which causes blister formation. Presented at the 77th Annual Meeting of the Federation of Societies for Coatings Technology, on October 18–22, 1999, in Dallas, TX. Dept. of Materials Engineering, Via Mesiano 77, Trento, Italy. e-mail: Flavio.Deflorian@ing.unitn.it; Stefano.Rossi@ing.unitn.it; Bonora@ing.unitn.it. Dept. ICMMPM, Via Eudossiana, Rome, Italy. e-mail: Lorenzo.Fedrizzi@ing.unitn.it.     

Dilatometry: A Tool to Measure the Influence of Cooling Rate and Pressure on the Phase Behavior of Nucleated Polypropylene

The pressure and cooling rate dependence of the phase diagram of isotactic polypropylene (iPP) with the nucleating agent 1,3:2,4‐bis(3,4‐dimethylbenzylidene)‐sorbitol (DMDBS) is investigated. A custom designed dilatometer is used to measure the specific volume of the blends for a wide range of cooling rates and elevated pressures. The crystallization line in the phase diagram shifts to higher temperatures with increase in the pressure and decrease in the cooling rate, independent of the concentration. The influence of cooling rate and pressure is related to the final morphology determined from X‐ray diffraction. Dilatometry can be used as an interesting alternative to extract information on the phase behavior and crystallinity, for conditions hard or not at all obtainable with standard techniques like DSC or SALS.

     

Direct and Indirect Effect of TGFβ on Treg Transendothelial Recruitment in HCC Tissue Microenvironment

The balance between anti-tumor and tumor-promoting immune cells, such as CD4+ Th1 and regulatory T cells (Tregs), respectively, is assumed to dictate the progression of hepatocellular carcinoma (HCC). The transforming growth factor beta (TGFβ) markedly shapes the HCC microenvironment, regulating the activation state of multiple leukocyte subsets and driving the differentiation of cancer associated fibroblasts (CAFs). The fibrotic (desmoplastic) reaction in HCC tissue strongly depends on CAFs activity. In this study, we attempted to assess the role of TGFβ on transendothelial migration of Th1-oriented and Treg-oriented CD4+ T cells via a direct or indirect, CAF-mediated mechanisms, respectively. We found that the blockage of TGFβ receptor I-dependent signaling in Tregs resulted in impaired transendothelial migration (TEM) of these cells. Interestingly, the secretome of TGFβ-treated CAFs inhibited the TEM of Tregs but not Th1 cells, in comparison to the secretome of untreated CAFs. In addition, we found a significant inverse correlation between alpha-SMA and FoxP3 (marker of Tregs) mRNA expression in a microarray analysis involving 78 HCCs, thus suggesting that TGFβ-activated stromal cells may counteract the trafficking of Tregs into the tumor. The apparent dual behavior of TGFβ as both pro- and anti-tumorigenic cytokines may add a further level of complexity to the mechanisms that regulate the interactions among cancerous, stromal, and immune cells within HCC, as well as other solid tumors, and contribute to better manipulation of the TGFβ signaling as a therapeutic target in HCC patients.