Simulating Assembly Landscapes for Comprehensive Understanding of Supramolecular Polymer-Solvent Systems

Complexity in supramolecular polymer systems arises from interactions between different components, including solvent molecules. By varying their concentration or temperature in such multicomponent systems, complex phenomena can occur such as thermally bisignate and dilution-induced assembly of supramolecular polymers. Herein, we demonstrate that both these phenomena emerge from the same underlying interaction mechanism between the components. As a model system, amide-decorated supramolecular polymers of porphyrins were investigated in combination with aliphatic alcohols as hydrogen-bond scavengers, and thermodynamic mass-balance models were applied to map the three-dimensional assembly landscapes. These studies unveiled that the interaction between hydrogen-bond scavengers and monomers is temperature-dependent and becomes dominant at high monomer concentrations. With these insights, we could exploit competitive monomer-alcohol interactions to prompt the dilution-induced assembly of various common monomers as well as bisignate assembly events. Moreover, kinetic insights were obtained by navigating through the assembly landscape. Similar to phase diagrams of covalent polymers, these assembly landscapes provide a comprehensive picture of supramolecular polymerizations, which helps to precisely regulate the system properties. The generality of this approach using assembly landscapes makes it relevant for any supramolecular system, and this enhanced control will open the door to build complex and functional supramolecular polymer systems.</p

25 学校ボランティア通信 No.13

発行日：2009年7月15

A Serine Palmitoyltransferase Inhibitor Blocks Hepatitis C Virus Replication in Human Hepatocytes

Background & AimsHost cell lipid rafts form a scaffold required for replication of hepatitis C virus (HCV). Serine palmitoyltransferases (SPTs) produce sphingolipids, which are essential components of the lipid rafts that associate with HCV nonstructural proteins. Prevention of the de novo synthesis of sphingolipids by an SPT inhibitor disrupts the HCV replication complex and thereby inhibits HCV replication. We investigated the ability of the SPT inhibitor NA808 to prevent HCV replication in cells and mice.MethodsWe tested the ability of NA808 to inhibit SPT’s enzymatic activity in FLR3-1 replicon cells. We used a replicon system to select for HCV variants that became resistant to NA808 at concentrations 4- to 6-fold the 50% inhibitory concentration, after 14 rounds of cell passage. We assessed the ability of NA808 or telaprevir to inhibit replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in mice with humanized livers (transplanted with human hepatocytes). NA808 was injected intravenously, with or without pegylated interferon alfa-2a and HCV polymerase and/or protease inhibitors.ResultsNA808 prevented HCV replication via noncompetitive inhibition of SPT; no resistance mutations developed. NA808 prevented replication of all HCV genotypes tested in mice with humanized livers. Intravenous NA808 significantly reduced viral load in the mice and had synergistic effects with pegylated interferon alfa-2a and HCV polymerase and protease inhibitors.ConclusionsThe SPT inhibitor NA808 prevents replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in cultured hepatocytes and in mice with humanized livers. It might be developed for treatment of HCV infection or used in combination with pegylated interferon alfa-2a or HCV polymerase or protease inhibitors

Motor planning error in Parkinson's disease and its clinical correlates.

This study aimed to investigate (a) motor planning difficulty by using a two-step test in Parkinson's disease (PD) compared with age-matched healthy subjects and (b) the relationship between motor planning difficulty and clinical factors. The two-step test was performed by 58 patients with PD with Hoehn & Yahr (H&Y) stage I-IV and 110 age-matched healthy older adult controls. In the two-step test, the participants estimated the two-step distance with maximum effort. Subsequently, they performed the actual two-step trial to measure the actual maximum distance. We calculated the accuracy of the estimation (estimated distance minus actual distance). In both groups, subjects who estimated >5 cm were defined as the overestimation group, and those who estimated <5 cm over- and underestimation were defined as the non-overestimation group. The overestimation group consisted of 17 healthy older adults (15.5%) and 23 patients with PD (39.7%). The number of patients with PD with overestimation was significantly more than that of healthy controls by Chi-squared test. H&Y stage and the Unified Parkinson's Disease Rating Scale (UPDRS) part II and III scores in overestimation group in PD patients were significantly higher than those in overestimation group in PD patients. Moreover, multiple regression using H&Y stage and UPDRS parts II and III as independent variables showed that the UPDRS part II score was the only related factor for the estimation error distance. Estimation error distance was significant correlated with UPDRS parts II and III. Patients with PD easily have higher rates of motor-related overestimation than age-matched healthy controls. In addition, UPDRS parts II and III expressed ability of activities of daily living and motor function as influences on motor-related overestimation. Particularly, multiple regression indicated that UPDRS part II directly showed the ability of daily living as an essential factor for overestimation

Bispecific Antibodies Produced via Chemical Site-Specific Conjugation Technology: AJICAP Second Generation

Bispecific antibodies are biotherapeutics that amalgamate the specificities of two distinct antibodies into one molecule. Bispecific antibodies can be utilized in a broad range of diagnostic and therapeutic applications; however, their engineering requires genetic modification and remains time-consuming. Therefore, in this study, we used AJICAP second-generation technology, which drives the production of site-specific antibody-drug conjugates in a practical and robust manner, without genetic modification requirements, to generate bispecific antibodies. Using haloketone chemistry as an alternative to maleimide chemistry, which carries reaction risks, we successfully produced site-specific antibody conjugates. Pharmacokinetic studies revealed that the haloketone-based antibody conjugate was stable in the rat plasma. The resultant bispecific antibodies were rigorously evaluated, and surface plasmon resonance measurements and flow cytometry analyses confirmed that antigen binding remained intact. Additionally, the affinity for the neonatal Fc receptor (FcRn) was retained after conjugation. Further cytotoxicity evaluation emphasized the pronounced activity of the generated bi-specific antibodies. These preliminary findings highlight the potential of AJICAP second-generation technology in BisAb production. This novel approach introduces a fully chemical, site-specific strategy capable of producing bispecific antibodies, heralding a new era in the field of biotherapeutics