Conjugation of PolyPEG to interferon alpha extends serum half-life while maintaining low viscosity of the conjugate.

The covalent attachment of poly(ethylene glycol) (PEG) to therapeutic proteins is a commonly used approach for extending in vivo half-lives. A potential limitation of this PEGylation strategy is the adverse effect of PEG on conjugate viscosity. Interferon-alpha (IFN) was conjugated via its N-terminal amino group by reductive amination to α-aldehyde functional comb-shaped PolyPEG polymers (50 and 70 kDa) and to linear PEG (30 kDa). In vitro potencies of the purified PEGylated IFN conjugates were measured by reporter gene assay using a HEK293P/ISRE-SEAP cell line. IFN levels were measured in rats following intravenous injection. Viscosities of various linear PEG and PolyPEG polymers along with the polymer-IFN conjugates were determined using a rotational rheometer with cone-and-plate geometry. In vitro potencies and half-lives of the PEGylated IFN conjugates were compared with those of the marketed branched PEG-IFN conjugate PEGASYS. Both PolyPEG-IFN conjugates retained a similar potency as that of the marketed comparator, whereas the linear PEG-IFN conjugate potency was greater. All conjugates showed extended half-lives compared to that of naked IFN, with the PolyPEG conjugates exhibiting the longest half-lives and the linear PEG conjugate, the shortest. Viscosity analysis showed that the linear PEG-IFN conjugate was over twice as viscous as both PolyPEG conjugates. Taken together, this work demonstrates the potential of PolyPEG conjugation to therapeutic proteins as a novel tool for optimizing pharmacokinetic profiles in a way that potentially allows administration of high-dose formulations because of lower conjugate viscosity.

Antifungal activity of cinnamic acid derivatives involves inhibition of benzoate 4-hydroxylase (CYP53).

AIMS: CYP53A15, from the sorghum pathogen Cochliobolus lunatus, is involved in detoxification of benzoate, a key intermediate in aromatic compound metabolism in fungi. Because this enzyme is unique to fungi, it is a promising drug target in fungal pathogens of other eukaryotes. METHODS AND RESULTS: In our work, we showed high antifungal activity of seven cinnamic acid derivatives against C. lunatus and two other fungi, Aspergillus niger and Pleurotus ostreatus. To elucidate the mechanism of action of cinnamic acid derivatives with the most potent antifungal properties, we studied the interactions between these compounds and the active site of C. lunatus cytochrome P450, CYP53A15. CONCLUSION: We demonstrated that cinnamic acid and at least four of the 42 tested derivatives inhibit CYP53A15 enzymatic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: By identifying selected derivatives of cinnamic acid as possible antifungal drugs, and CYP53 family enzymes as their targets, we revealed a potential inhibitor-target system for antifungal drug development.

The cost of attack in competing networks.

Real-world attacks can be interpreted as the result of competitive interactions between networks, ranging from predator-prey networks to networks of countries under economic sanctions. Although the purpose of an attack is to damage a target network, it also curtails the ability of the attacker, which must choose the duration and magnitude of an attack to avoid negative impacts on its own functioning. Nevertheless, despite the large number of studies on interconnected networks, the consequences of initiating an attack have never been studied. Here, we address this issue by introducing a model of network competition where a resilient network is willing to partially weaken its own resilience in order to more severely damage a less resilient competitor. The attacking network can take over the competitor's nodes after their long inactivity. However, owing to a feedback mechanism the takeovers weaken the resilience of the attacking network. We define a conservation law that relates the feedback mechanism to the resilience dynamics for two competing networks. Within this formalism, we determine the cost and optimal duration of an attack, allowing a network to evaluate the risk of initiating hostilities.

Effect of hydralazine on interstitial fluid pressure in experimental tumours and in normal tissue.

Interstitial fluid pressure (IFP) has been recognised as the most important obstacle in macromolecular drug delivery to solid tumours. The aim of our study was to measure the IFP simultaneously in tumour and in muscle or in subcutis and to determine whether injection of hydralazine reduces differentially tumour IFP with respect to IFP in surrounding and normal tissues. In addition, it was of interest whether the decrease in IFP due to hydralazine depends on tumour volume and/or on initial IFP. Measurements of IFP were performed by means of the wick-in-needle technique and they were obtained on tumours of different size. In both tumour models, hydralazine significantly reduced the pretreatment IFP level. On average IFP decreased by 31% and 14% from the initial value in SAF and LPB tumours, respectively. On the contrary, hydralazine did not decrease IFP in normal tissue. Injection of NaCl solution instead of hydralazine had no effect on IFP either in tumours or in subcutis/muscle. The results of our study on the effect of hydralazine on IFP in SAF and LPB tumour model are in accordance to previously reported studies. The initial IFP in tumour is positively-correlated with the tumour size, while the decrease in the tumour IFP is independent of the initial IFP value. In addition, the decrease in tumour IFP is not correlated to tumour volume.

Network risk and forecasting power in phase-flipping dynamical networks.

To model volatile real-world network behavior, we analyze a phase-flipping dynamical scale-free network in which nodes and links fail and recover. We investigate how stochasticity in a parameter governing the recovery process affects phase-flipping dynamics, and we find the probability that no more than q% of nodes and links fail. We derive higher moments of the fractions of active nodes and active links, fn(t) and fℓ(t), and we define two estimators to quantify the level of risk in a network. We find hysteresis in the correlations of fn(t) due to failures at the node level, and we derive conditional probabilities for phase-flipping in networks. We apply our model to economic and traffic networks.

Semi-preparative chromatographic purification of the enantiomers S-(-)-amlodipine and R-(+)-amlodipine.

Pharmacokinetic studies of optically pure compounds after single enantiomer administration are becoming increasingly important. The process of racemization in vivo can diminish all expected advantages of single enantiomer treatment. Amlodipine, one of the calcium channel blockers, currently used in therapy as a racemate, is one of such drugs under study. In order to administer single enantiomers of amlodipine to healthy volunteers both were chromatographically purified and characterised. The two optical isomers of amlodipine, active S-(-)- and non-active R-(+)-amlodipine, were purified using chromatographic procedure adopted from the analytical separation. Enantiomers were successfully converted to benzenesulphonic salt without any racemization. All semi-preparative purifications were monitored with complementary analytical methods, HPLC and CE, along with the determination of optical activity so that the final product was sufficiently defined for further in vivo studies. The analytical method developed for the determination of plasma concentrations of each enantiomer of amlodipine in these studies is also briefly described.