Discovery of Affinity-Based Probes for Btk Occupancy Assays

Bruton's tyrosine kinase (Btk) is an attractive target for the treatment of a wide array of B-cell malignancies and autoimmune diseases. Small-molecule covalent irreversible Btk inhibitors targeting Cys481 have been developed for the treatment of such diseases. In clinical trials, probe molecules are required in occupancy studies to measure the level of engagement of the protein by these covalent irreversible inhibitors. The result of this pharmacodynamic (PD) activity provides guidance for appropriate dosage selection to optimize inhibition of the drug target and correlation of target inhibition with disease treatment efficacy. This information is crucial for successful evaluation of drug candidates in clinical trials. Based on the pyridine carboxamide scaffold of a novel solvent-accessible pocket (SAP) series of covalent irreversible Btk inhibitors, we successfully developed a potent and selective affinity-based biotinylated probe 12 (2-[(4-{4-[5-(1-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-3,6,9,12-tetraoxapentadecan-15-amido)pentanoyl]piperazine-1-carbonyl}phenyl)amino]-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide). Compound 12 has been used in Btk occupancy assays for preclinical studies to determine the therapeutic efficacy of Btk inhibition in two mouse lupus models driven by TLR7 activation and type I interferon.     

双波段比色辐射温度计的波长选择

通过对比色测温法灵敏度温区特性和波比数判据的分析,讨论了比色波长的匹配原理,试图解决选择比色波长的理论依据。     

On the Yustova threshold plates

A response to criticism of threshold plates for the study of color vision developed at the Mendeleev All-Russia Research Institute of Metrology and published in 1994 is presented. The critics base their conclusions on colorimetric testing and the examination of the plates. In response to the article by M. V. Danilova and J. D. Mollon [4]. __________ Translated from Izmeritel’naya Tekhnika, No. 1, pp. 37–39, January, 2007.     

速溶奶粉中卵磷脂含量的测定

叙述了速溶奶粉中卵磷脂的测定方法，样品通过一系列处理后，测定其磷含量，回收率为９６％￣９８％。     

Chromatic luminance,colorimetric purity,and optimal aperture‐color stimuli

Chromatic luminance (i.e., luminance of a monochromatic color) is the source of all luminance, since achromatic luminance arises only from mixing colors and their chromatic luminances. The ratio of chromatic luminance to total luminance (i.e., chromatic plus achromatic luminance) is known as colorimetric purity, and its measurement has long been problematic for nonspectral hues. Colorimetric purity (p_c) is a luminance metric in contrast to excitation purity, which is a chromaticity‐diagram metric approximating saturation. The CIE definition of p_c contains a fallacy. CIE defines maximum (1.0) p_c for spectral stimuli as monochromatic (i.e., optimal) stimuli, and as the line between spectrum ends for nonspectrals. However, this line has <0.003 lm/W according to CIE colorimetric data and is therefore effectively invisible. It only represents the limit of theoretically attainable colors, and is of no practical use in color reproduction or color appearance. Required is a locus giving optimal rather than invisible nonspectral stimuli. The problem is partly semantic. CIE wisely adopted the term colorimetric purity, rather than the original spectral luminance purity, to permit an equivalent metric for spectrals and nonspectrals, but the parameter of equivalence was never clear. Since 1 p_c denotes optimal aperture‐color stimuli for spectrals, arguably 1 p_c should denote optimal stimuli consistently for all stimuli. The problem reduces to calculating optimal aperture‐color stimuli (“optimal” in energy efficiency in color‐matching) for nonspectrals, shown to comprise 442 + 613 nm in all CIE illuminants. This remedy merely requires redefinition of 1 p_c for nonspectrals as the line 442–613 nm, and gives meaningful p_c values over the hue cycle allowing new research of chromatic luminance relations with color appearance. © 2007 Wiley Periodicals, Inc. Col Res Appl, 32, 469–476, 2007     

彩色CCD比色测温的灰度值融合处理方法研究

设计了基于两个彩色CCD通道的比色测温系统,提出了用于比色测温时彩色CCD的R、G、B亮度(灰度)值融合处理方法,解决了在不同快门下利用两个通道的灰度值进行比色测温计算的问题;黑体炉实验结果表明,与常用的基于面阵CCD的简化比色测温方法相比较,基于两个彩色CCD通道的比色测温方法系统虽稍显复杂,但由于采用了经融合处理后的亮度值进行比色测量计算使得测温精度较高。     

Human DDX3X Unwinds Japanese Encephalitis and Zika Viral 5′ Terminal Regions

Flavivirus genus includes many deadly viruses such as the Japanese encephalitis virus (JEV) and Zika virus (ZIKV). The 5′ terminal regions (TR) of flaviviruses interact with human proteins and such interactions are critical for viral replication. One of the human proteins identified to interact with the 5′ TR of JEV is the DEAD-box helicase, DDX3X. In this study, we in vitro transcribed the 5′ TR of JEV and demonstrated its direct interaction with recombinant DDX3X (K_d of 1.66 ± 0.21 µM) using microscale thermophoresis (MST). Due to the proposed structural similarities of 5′ and 3′ TRs of flaviviruses, we investigated if the ZIKV 5′ TR could also interact with human DDX3X. Our MST studies suggested that DDX3X recognizes ZIKV 5′ TR with a K_d of 7.05 ± 0.75 µM. Next, we performed helicase assays that suggested that the binding of DDX3X leads to the unwinding of JEV and ZIKV 5′ TRs. Overall, our data indicate, for the first time, that DDX3X can directly bind and unwind in vitro transcribed flaviviral TRs. In summary, our work indicates that DDX3X could be further explored as a therapeutic target to inhibit Flaviviral replication     

Characterization of New DNA Aptamers for Anti-HIV-1 Reverse Transcriptase

HIV-1 RT is a necessary enzyme for retroviral replication, which is the main target for antiviral therapy against AIDS. Effective anti-HIV-1 RT drugs are divided into two groups; nucleoside inhibitors (NRTI) and non-nucleoside inhibitors (NNRTI), which inhibit DNA polymerase. In this study, new DNA aptamers were isolated as anti-HIV-1 RT inhibitors. The selected DNA aptamer (WT62) presented with high affinity and inhibition against wild-type (WT) HIV-1 RT and gave a KD value of 75.10±0.29 nM and an IC₅₀ value of 84.81±8.54 nM. Moreover, WT62 decreased the DNA polymerase function of K103 N/Y181 C double mutant (KY) HIV-1 RT by around 80 %. Furthermore, the ITC results showed that this aptamer has small binding enthalpies with both WT and KY HIV-1 RTs through which the complex might form a hydrophobic interaction or noncovalent bonding. The NMR result also suggested that the WT62 aptamer could bind with both WT and KY mutant HIV-1 RTs at the connection domain.     

硼矿制硼酸母液中硼含量的测定方法探讨

报道了笔者用水相直接显色型比色法测定硼酸母液中硼含量的实验和结果。