General description

The GSK-J4, with the CAS No: 1373423-53-0, is also known as β-Alanine,N-[2-(2-pyridinyl)-6-(1,2,4,5-tetrahydro-3H-3-benzazepin-3-yl)-4-pyrimidinyl]-, ethylester. This chemical’s molecular formula is C24H27N5O2 and molecular weight is 417.504. It is a potent H3K27me3 demethylase inhibitor.[1]

Figure 1 the molecular formula of GSK-J4

Indication

In 2013, the first selective and potent inhibitor of JMJD3/UTX demethyltransferase activity, GSK-J4, was synthesized by GlaxoSmithKline. Subsequently, two studies have addressed the effect of this drug in immune cells. A study reported that GSK-J4 attenuates lipopolysaccharide (LPS)induced proinflammatory cytokine production in primary human macrophages in vitro.[2] The another study addressed the effects of this drug in T-cell differentiation in vitro and that Jmjd3-deficient mice were resistant to the induction of experimental autoimmune encephalomyelitis (EAE). However, the efficacy of GSK-J4 in inflammation in vivo in the context of autoimmunity has not yet been addressed. Thus, this information persuaded research worker to study the effect of GSK-J4 in a CD4t T-cell mediated response in vivo and its relevance in autoimmunity using a murine model of multiple sclerosis (MS).[3]

Pharmacodynamics

GSK-J4 is a potent dual inhibitor of H3K27me3/me2-demethylases JMJD3/KDM6B and UTX/KDM6A with IC50s of 8.6 and 6.6 μM, respectively. It inhibits LPS-induced TNF-α production in human primary macrophages with an IC50 of 9 μM. GSK J4 is a cell permeable prodrug of GSK-J1[5-7]. GSK-J4 induces endoplasmic reticulum stress-related apoptosis[8].  GSK-J4 (0.5 mg/kg, i.p.) significantly reduces the severity and delays the onset of the disease of the mouse model of experimental autoimmune encephalomyelitis[9]. 

In Vitro:As it has been established that demethylation of lysine 27 of histone H3 by the lysine-specific demethylase JMJD3 increases immune responses and thus elicits inflammation, we found that in vivo administration of GSK-J4, a selective inhibitor of JMJD3 and UTX, ameliorates the severity of experimental autoimmune encephalomyelitis (EAE). In vitro experiments revealed that the anti-inflammatory effect of GSK-J4 was exerted through an effect on dendritic cells (DCs), promoting a tolerogenic profile characterized by reduced expression of costimulatory molecules CD80/CD86, an increased expression of tolerogenic molecules CD103 and TGF-β1, and reduced secretion of proinflammatory cytokines IL-6, IFN-γ, and TNF. Adoptive transfer of GSK-J4-treated DCs into EAE mice reduced the clinical manifestation of the disease and decreased the extent of inflammatory CD4+ T cells infiltrating the central nervous system. Notably, Treg generation, stability, and suppressive activity were all exacerbated by GSK-J4-treated DCs without affecting Th1 and Th17 cell production. Our data show that GSK-J4-mediated modulation of inflammation is achieved by a direct effect on DCs and that systemic treatment with GSK-J4 or adoptive transfer of GSK-J4-treated DCs ex vivo may be promising approaches for the treatment of inflammatory and autoimmune disorders.[7]

In Vivo:GSK-J4 has cellular activity in Flag-JMJD3-transfected HeLa cells, in which GSK-J4 prevents the JMJD3-induced loss of nuclear H3K27me3 immunostaining.Administration of GSK-J4 increases total nuclear H3K27me3 levels in untransfected cells. GSK-J4 significantly reduces the expression of 16 of 34 LPS-driven cytokines, including tumour-necrosis factor-α (TNFα)[2]. GSK-J4 (5 μM; 48 hours) causes a more than 3-fold increase in mouse podocyte H3K27me3 content. H3K27me3 levels in cultured podocytes, GSK-J4 reduces Jagged-1 mRNA and Jagged-1 protein levels. Correspondingly, when exposed podocytes to the inducer of dedifferentiation TGF-β1, pretreatment with GSK-J4 preventes both the increase in intracellular N1-ICD levels and the increase in α-SMA and the decrease in podocin mRNA levels[1]. GSK-J4 (10, 25 nM) acts upon DCs promoting the differentiation of Treg cells, improving Treg stability and suppressive capacities, without affecting the differentiation of Th1 and Th17 cells[7]. GSK-J4 inhibits JMJD3 expression that is induced by TGF-β1.[8] GSK-J4 inhibits H3K4 demethylation at Xist, Nodal, and HoxC13 in female embryonic stem cells.[9] MCE has not independently confirmed the accuracy of these methods. They are for reference only. GSK-J4 Hydrochloride (10 mg/kg; i.p.; thrice-weekly for 10 weeks) attenuates the development of kidney disease in diabetic mice.[2] GSK-J4 (0.5 mg/kg, i.p.) significantly reduces the severity and delays the onset of the disease of the mouse model of experimental autoimmune encephalomyelitis.[3]. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Based on previous studies demonstrating the in vitro potential of GSK-J4 to restrain the inflammatory response of macrophages [2], to affect T-cell differentiation and that Jmjd3-deficient mice were resistant to the induction of experimental autoimmune encephalomyelitis (EAE) .Using the mouse model of MS, the EAE[3], and immunized C57BL/6 wild-type mice with MOG3555 peptide emulsified in CFA, followed by daily intraperitoneal injections (i.p.) of GSK-J4 or vehicle for 5 days after EAE induction, and compared the susceptibility of untreated and GSK-J4-treated mice to develop the disease. The results show that administration of GSK-J4 significantly reduced the severity of the disease when compared with control mice (maximum severity of 2.13 ± 1.13 in GSK-J4-treated mice versus 4.25 ± 1.04 in control mice. Additionally, GSK-J4 treatment delayed the onset of the disease compared with untreated mice (onset of 18.14 ± 2.79 in GSK-J4-treated mice versus 15.50 ± 2.27 in control mice. In addition to the post-induction treatment of EAE with GSK-J4, researchers also tested the therapeutic potential of this drug by administrating it in a postonset regime. For this purpose, they injected GSK-J4 or the vehicle i.p. daily for five consecutive days starting the day after the onset. Strikingly, the results show that even when administrated after the onset, GSK-J4 exerted a therapeutic effect in EAE manifestation. These data demonstrate that systemic administration of a highly selective inhibitor of H3K27m3 demethylases exerts a therapeutic effect attenuating EAE development and progression.[3]

Mechanism of action

By systemic treatment with GSK-J4 or adoptive transfer of GSK-J4-treated DCs attenuated the inflammatory response mediated by CD4t T cells and the manifestation of EAE.[4]

References

[1]Heinemann B, et al. Inhibition of demethylases by GSK-J1/J4[J]. Nature. 2014, 2,514(7520):E1-2.

[2] Kruidenier L, et al. A selective jumonji H3K27 demethylase inhibitor modulates the proinflammatory macrophage response[J]. Nature. 2012, 16, 488 (7411):404-8. [3]Z. Liu, W. Cao, L. Xu, X. Chen, Y. Zhan, Q. Yang, et al. The histone H3 lysine-27 demethylase Jmjd3 plays a critical role in specific regulation of Th17 cell differentiation[J], Mol. Cell. Biol.2015:505-516.

[4]Cristian D, Macarena C, Macarena F, et al. The histone demethylase inhibitor GSK-J4 limits inflammation through the induction of a tolerogenic phenotype on DCs[J]. 2016.

[5]Kruidenier L, et al. A selective jumonji H3K27 demethylase inhibitor modulates the proinflammatory macrophage response. Nature. 2012, 16, 488(7411): 404-8.  

[6]Majumder S, et al. Shifts in podocyte histone H3K27me3 regulate mouse and human glomerular disease. J Clin Invest. 2018, 2, 128 (1): 483-499. 

[7]Donas C, et al. The histone demethylase inhibitor GSK-J4 limits inflammation through the induction of a tolerogenic phenotype on DCs[J]. Autoimmun. 2016, 75: 105-117.  

[8]Xuan Chu, et al. GSK-J4 induces cell cycle arrest and apoptosis via ER stress and the synergism between GSK-J4 and decitabine in acute myeloid leukemia KG-1a cells[J]. Cancer Cell International 2020, 20: 209.

[9]Yapp C, et al. H3K27me3 demethylases regulate in vitro chondrogenesis and chondrocyte activity in osteoarthritis. Arthritis Res Ther. 2016, 7;18(1):158. 

[10]Kamikawa YF, et al. Histone demethylation maintains Prdm14 and Tsix expression and represses xIst in embryonic stem cells. PLoS One. 2015.