Discovery of Pyrazolo[1,5-a]pyrimidine Derivative as a Novel and Selective ALKBH5 Inhibitor for the Treatment of AML.

M6A (N6-methyladenosine) plays a significant role in regulating RNA processing, splicing, nucleation, translation, and stability. AlkB homologue 5 (ALKBH5) is an Fe(II)/2-oxoglutarate (2-OG)-dependent dioxygenase that demethylates mono- or dimethylated adenosines. ALKBH5 can be regarded as an oncogenic factor for various human cancers. However, the discovery of potent and selective ALKBH5 inhibitors remains a challenge. We identified DDO-2728 as a novel and selective inhibitor of ALKBH5 by structure-based virtual screening and optimization. DDO-2728 was not a 2-oxoglutarate analogue and could selectively inhibit the demethylase activity of ALKBH5 over FTO. DDO-2728 increased the abundance of m6A modifications in AML cells, reduced the mRNA stability of TACC3, and inhibited cell cycle progression. Furthermore, DDO-2728 significantly suppressed tumor growth in the MV4-11 xenograft mouse model and showed a favorable safety profile. Collectively, our results highlight the development of a selective probe for ALKBH5 that will pave the way for the further study of ALKBH5 targeting therapies.

Electrical exposure analysis of galvanic-coupled intra-body communication based on the empirical arm models.

BACKGROUND: Intra-body communication (IBC) is one of the highlights in studies of body area networks. The existing IBC studies mainly focus on human channel characteristics of the physical layer, transceiver design for the application, and the protocol design for the networks. However, there are few safety analysis studies of the IBC electrical signals, especially for the galvanic-coupled type. Besides, the human channel model used in most of the studies is just a multi-layer homocentric cylinder model, which cannot accurately approximate the real human tissue layer. METHODS: In this paper, the empirical arm models were established based on the geometrical information of six subjects. The thickness of each tissue layer and the anisotropy of muscle were also taken into account. Considering the International Commission on Non-Ionizing Radiation Protection (ICNIRP) guidelines, the restrictions taken as the evaluation criteria were the electric field intensity lower than 1.35 × 104 f V/m and the specific absorption rate (SAR) lower than 4 W/kg. The physiological electrode LT-1 was adopted in experiments whose size was 4 × 4 cm and the distance between each center of adjoining electrodes was 6 cm. The electric field intensity and localized SAR were all computed by the finite element method (FEM). The electric field intensity was set as average value of all tissues, while SAR was averaged over 10 g contiguous tissue. The computed data were compared with the 2010 ICNIRP guidelines restrictions in order to address the exposure restrictions of galvanic-coupled IBC electrical signals injected into the body with different amplitudes and frequencies. RESULTS: The input alternating signal was 1 mA current or 1 V voltage with the frequency range from 10 kHz to 1 MHz. When the subject was stimulated by a 1 mA alternating current, the average electric field intensity of all subjects exceeded restrictions when the frequency was lower than 20 kHz. The maximum difference among six subjects was 1.06 V/m at 10 kHz, and the minimum difference was 0.025 V/m at 400 kHz. While the excitation signal was a 1 V alternating voltage, the electric field intensity fell within the exposure restrictions gradually as the frequency increased beyond 50 kHz. The maximum difference among the six subjects was 2.55 V/m at 20 kHz, and the minimum difference was 0.54 V/m at 1 MHz. In addition, differences between the maximum and the minimum values at each frequency also decreased gradually with the frequency increased in both situations of alternating current and voltage. When SAR was introduced as the criteria, none of the subjects exceeded the restrictions with current injected. However, subjects 2, 4, and 6 did not satisfy the restrictions with voltage applied when the signal amplitude was ≥ 3, 6, and 10 V, respectively. The SAR differences for subjects with different frequencies were 0.062-1.3 W/kg of current input, and 0.648-6.096 W/kg of voltage input. CONCLUSION: Based on the empirical arm models established in this paper, we came to conclusion that the frequency of 100-300 kHz which belong to LF (30-300 kHz) according to the ICNIRP guidelines can be considered as the frequency restrictions of the galvanic-coupled IBC signal. This provided more choices for both intensities of current and voltage signals as well. On the other hand, it also makes great convenience for the design of transceiver hardware and artificial intelligence application. With the frequency restrictions settled, the intensity restrictions that the current signal of 1-10 mA and the voltage signal of 1-2 V were accessible. Particularly, in practical application we recommended the use of the current signals for its broad application and lower impact on the human tissue. In addition, it is noteworthy that the coupling structure design of the electrode interface should attract attention.