Development of Kinase-Selective, Harmine-Based DYRK1A Inhibitors that Induce Pancreatic Human β-Cell Proliferation

Abstract

DYRK1A has been implicated as an important drug target in various therapeutic areas, including neurological disorders and oncology. DYRK1A has more recently been shown to be involved in pathways regulating human β-cell proliferation, thus making it a potential therapeutic target for both Type 1 and Type 2 diabetes. Our group, using a high-throughput phenotypic screen, identified harmine that is able to induce β-cell proliferation both in vitro and in vivo. Since harmine has suboptimal kinase selectivity, we sought to expand structure-activity relationships for harmine's DYRK1A activity, to enhance selectivity, while retaining human β-cell proliferation capability. We carried out the optimization of the 1-position of harmine and synthesized 15 harmine analogues. Six compounds showed excellent DYRK1A inhibition with IC in the range of 49.5-264 nM. Two compounds, 2-2 and 2-8, exhibited excellent human β-cell proliferation at doses of 3-30 μM, and compound 2-2 showed improved kinase selectivity as compared to harmine.

Pharmacologic and genetic approaches define human pancreatic β cell mitogenic targets of DYRK1A inhibitors

“…In the past 4 years, several groups have shown that drugs that inhibit the β cell kinase, dual specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) are able to induce proliferation of human β cells in vitro and in vivo. This class of human β cell proliferation-enhancing DYRK1A inhibitors includes harmine, INDY, leucettine-41, GNF4877, 5-iodotubericidin (5-IT), TG003, AZ191, CC-401, and more recently synthesized DYRK1A inhibitors (5)(6)(7)(8)(9)(10)(11)(12)(13). Several reports have shown that the human β cell proliferative activity of this class can be mimicked by silencing DYRK1A and can be inhibited by overexpression of DYRK1A in human β cells (5-7), making it clear that DYRK1A is an essential mediator of the proliferative response to these drugs.…”

Human Beta Cell Mass Expansion In Vivo With A Harmine and Exendin-4 Combination: Quantification and Visualization By iDISCO+ 3D Imaging

“…Chemicals. Harmine-HCl was synthesized in the Drug Discovery Institute at Mount Sinai (15,16). Exendin-4 was purchased from MedChemExpress (Monmouth Junction, NJ).…”

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

“…Compounds were tested for DYRK1A binding activity by a commercial kinase profiling services, Life Technologies which uses the FRET-based LanthaScreen ® Eu Kinase Binding Assay. [38] Compounds were screened for DYRK1A activity at concentrations of 1000 nM and 300 nM in duplicates. The IC 50 was determined by 10 point LanthaScreen ® Eu Kinase Binding Assay [38] in duplicates.…”

“…[38] Compounds were screened for DYRK1A activity at concentrations of 1000 nM and 300 nM in duplicates. The IC 50 was determined by 10 point LanthaScreen ® Eu Kinase Binding Assay [38] in duplicates.…”

“…Based on the known binding pose of harmine, we surmised that there are three positions, 1-methyl,7-methoxy and 9-N indole, where rational modifications of harmine can be carried out ( Figure 1) without disrupting key DYRK1A binding contacts. Previously, we reported the optimization of the 1-position of harmine which led to two compounds with robust human β-cell proliferation at doses of 3-30 µ M and one of those, compound I (R = CH2OH), showing improved kinase selectivity as compared to harmine ( Figure 1) [38]. Next, we recently reported the optimization of the N-9 position to identify 2-2c, a highly optimized in vivo active harmine based DYRK1A inhibitor (Figure 1) [39].…”