Selective DYRK1A Inhibitor for the Treatment of Type 1 Diabetes: Discovery of 6-Azaindole Derivative GNF2133

Abstract

Autoimmune deficiency and destruction in either βcell mass or function can cause insufficient insulin levels and, as a result, hyperglycemia and diabetes. Thus, promoting β-cell proliferation could be one approach toward diabetes intervention. In this report we describe the discovery of a potent and selective DYRK1A inhibitor GNF2133, which was identified through optimization of a 6-azaindole screening hit. In vitro, GNF2133 is able to proliferate both rodent and human β-cells. In vivo, GNF2133 demonstrated significant dose-dependent glucose disposal capacity and insulin secretion in response to glucosepotentiated arginine-induced insulin secretion (GPAIS) challenge in rat insulin promoter and diphtheria toxin A (RIP-DTA) mice. The work described here provides new avenues to disease altering therapeutic interventions in the treatment of type 1 diabetes (T1D).

Structure–Activity Relationships and Biological Evaluation of 7-Substituted Harmine Analogs for Human β-Cell Proliferation

“…Recently, our group found that harmine is able to induce human β-cell proliferation and DYRK1A-NFAT pathway as being the major pathway for this cell proliferation [5]. These results have been confirmed in other labs with other DYRK1A inhibitors unrelated to the harmine scaffold, including from our own lab [6,8,[34][35][36][37]. Since, modification of harmine has not been explored previously in context of β-cell proliferation, we therefore decided to carry out structure-activity relationship studies of harmine for both DYRK1A inhibition and β-cell proliferation.…”

Human Beta Cell Regenerative Drug Therapy for Diabetes: Past Achievements and Future Challenges

“…The first report of a DYRK1A inhibitor able to induce beta cell proliferation was reported in 2012 by Annes et al, who demonstrated that 5-iodotubericidin (5-IT) is able to induce rodent and porcine beta cells to replicate, an effect initially attributed to the ability of 5-IT to inhibit adenosine kinase (47). In 2015 through 2020, multiple groups including Laffite et al, Wagner et al, Annes et al, and ourselves showed that multiple DYRK1A inhibitors -harmine, INDY, leucettine-41, GNF4877, GNF2133, CC-401, OTS-167, and 2-2c -are able to induce human beta cells to replicate, as assessed by Ki67, BrdU, EdU, PHH3 immunolabeling, at rates of 2-3% (28,29,(47)(48)(49)(50)(51)(52)(53)(54)(55). Importantly, human beta cell proliferation can be reproduced by directly silencing DYRK1A gene expression in human islets (28,29,(48)(49)(50)(51).…”

“…DYRK1A is a kinase that phosphorylates a number of substrate proteins, among which are the Nuclear Factor activated in T-cells (NFaT) family of four transcription factors (31,48,(55)(56)(57). NFaTs normally reside in the cytoplasm, in a phosphorylated state.…”

“…It is clear that adult human beta cells are refractory to induction of proliferation, as summarized earlier: basal labeling indices in vitro and in vivo are in the 0.0-0.5% range as assessed using Ki67, PCNA, and 18-24 hours of BrdU or EdU exposure (13, 26-29, 33, 34, 45, 47-55, 62). While harmine and other DYRK1A inhibitors increase proliferation, this is only in the 2-4% range (28,(47)(48)(49)(50)(51)(52)(53)(54)(55), and DYRK1A inhibitors in combination treatment with GLP1 receptor agonists or TGFb inhibitors increase this only to 5-8% (28,29). This means that even under optimal circumstances, >90% of beta cells do not proliferate.…”

Down Syndrome Is a Metabolic Disease: Altered Insulin Signaling Mediates Peripheral and Brain Dysfunctions

“…Pharmacological inhibition of DYRK1A in both mice and human cells leads to proliferation of β-cell (Wang et al, 2015;Belgardt and Lammert, 2016) and improves glycemic control in mice (Liu et al, 2020). Dyrk1A is highly expressed and induces the expression and nuclear accumulation of p27Kip1 in the adipose tissue and pancreas (Rachdi et al, 2014b).…”