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HM71224, a novel Bruton's tyrosine kinase inhibitor, suppresses B cell and monocyte activation and ameliorates arthritis in a mouse model: a potential drug for rheumatoid arthritis.

Park JK, Byun JY, Park JA, Kim YY, Lee YJ, Oh JI, Jang SY, Kim YH, Song YW, Son J, Suh KH, Lee YM, Lee EB - Arthritis Res. Ther. (2016)

Bottom Line: The in vitro effects of HM71224 on B cells and monocytes were analyzed by examining phosphorylation of Btk and its downstream signaling molecules, along with cytokine production and osteoclast formation.Furthermore, HM71224 effectively inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β by human monocytes, and osteoclast formation by human monocytes.Thus, HM71224 is a potential novel therapeutic agent for rheumatoid arthritis in humans.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Department of Internal Medicine, Seoul National University College of Medicine, 101 Daehank-no Chongno-gu, Seoul, 03080, Korea.

ABSTRACT

Background: Bruton's tyrosine kinase (Btk) is critical for activation of B cells and myeloid cells. This study aimed to characterize the effects of HM71224, a novel Btk inhibitor, both in vitro and in a mouse model of experimental arthritis.

Methods: The kinase inhibition profile of HM71224 was analyzed. The in vitro effects of HM71224 on B cells and monocytes were analyzed by examining phosphorylation of Btk and its downstream signaling molecules, along with cytokine production and osteoclast formation. The in vivo effects of HM71224 were investigated in a mouse model of collagen-induced arthritis (CIA).

Results: HM71224 irreversibly bound to and inhibited Btk (IC50 = 1.95 nM). The compound also inhibited the phosphorylation of Btk and its downstream molecules such as PLCγ2, in activated Ramos B lymphoma cells and primary human B cells in a dose-dependent manner. Furthermore, HM71224 effectively inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β by human monocytes, and osteoclast formation by human monocytes. Finally, HM71224 improved experimental arthritis and prevented joint destruction in a murine model of CIA.

Conclusions: HM71224 inhibits Btk in B cells and monocytes and ameliorates experimental arthritis in a mouse model. Thus, HM71224 is a potential novel therapeutic agent for rheumatoid arthritis in humans.

No MeSH data available.


Related in: MedlinePlus

HM71224 inhibits cytokine production by human peripheral blood monocytes and plasmacytoid dendritic cells. a Human CD14+ monocytes were isolated from healthy donors (n = 3) and stimulated with heat-inactivated immune complexes (ICs) in the presence of HM71224. The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 5 h post stimulation. HM71224 significantly reduced the levels of secreted cytokines in a dose-dependent manner. b Human peripheral blood dendritic cells were isolated from healthy donors (n = 2) and stimulated with CpG oligodeoxynucleotides (CpG ODN) or lipopolysaccharide (LPS). The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 48 h post stimulation.*P < 0.05, **P < 0.01, and ***P < 0.001 vs. no treatment. LPS lipopolysaccharide
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Fig4: HM71224 inhibits cytokine production by human peripheral blood monocytes and plasmacytoid dendritic cells. a Human CD14+ monocytes were isolated from healthy donors (n = 3) and stimulated with heat-inactivated immune complexes (ICs) in the presence of HM71224. The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 5 h post stimulation. HM71224 significantly reduced the levels of secreted cytokines in a dose-dependent manner. b Human peripheral blood dendritic cells were isolated from healthy donors (n = 2) and stimulated with CpG oligodeoxynucleotides (CpG ODN) or lipopolysaccharide (LPS). The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 48 h post stimulation.*P < 0.05, **P < 0.01, and ***P < 0.001 vs. no treatment. LPS lipopolysaccharide

Mentions: Btk is involved in signal transduction by FcγRs expressed by myeloid lineage cells such as monocytes, macrophages, and neutrophils [15]. Therefore, we next examined the effect of HM71224 on inflammatory cytokine production by human monocytes. Human CD14+ cells were isolated from healthy volunteers and stimulated with heat-inactivated ICs in the presence of increasing concentrations of HM71224. The production of TNF-α, IL-1β, and IL-6 was inhibited in a dose-dependent manner (Fig. 4a). Btk also plays a crucial role in activating human PDCs via the TLR9 signaling pathway [16]. As expected, stimulation of human PDCs with either LPS (a TLR4 agonist) or CpG oligodeoxynucleotides (CpG ODN, a TLR9 agonist) induced the production of TNF-α and IL-6. HM71224 inhibited CpG-mediated cytokine production but not that mediated by LPS (Fig. 4b).Fig. 4


HM71224, a novel Bruton's tyrosine kinase inhibitor, suppresses B cell and monocyte activation and ameliorates arthritis in a mouse model: a potential drug for rheumatoid arthritis.

Park JK, Byun JY, Park JA, Kim YY, Lee YJ, Oh JI, Jang SY, Kim YH, Song YW, Son J, Suh KH, Lee YM, Lee EB - Arthritis Res. Ther. (2016)

HM71224 inhibits cytokine production by human peripheral blood monocytes and plasmacytoid dendritic cells. a Human CD14+ monocytes were isolated from healthy donors (n = 3) and stimulated with heat-inactivated immune complexes (ICs) in the presence of HM71224. The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 5 h post stimulation. HM71224 significantly reduced the levels of secreted cytokines in a dose-dependent manner. b Human peripheral blood dendritic cells were isolated from healthy donors (n = 2) and stimulated with CpG oligodeoxynucleotides (CpG ODN) or lipopolysaccharide (LPS). The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 48 h post stimulation.*P < 0.05, **P < 0.01, and ***P < 0.001 vs. no treatment. LPS lipopolysaccharide
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835877&req=5

Fig4: HM71224 inhibits cytokine production by human peripheral blood monocytes and plasmacytoid dendritic cells. a Human CD14+ monocytes were isolated from healthy donors (n = 3) and stimulated with heat-inactivated immune complexes (ICs) in the presence of HM71224. The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 5 h post stimulation. HM71224 significantly reduced the levels of secreted cytokines in a dose-dependent manner. b Human peripheral blood dendritic cells were isolated from healthy donors (n = 2) and stimulated with CpG oligodeoxynucleotides (CpG ODN) or lipopolysaccharide (LPS). The levels of the indicated cytokines in the culture supernatant were then measured by ELISA at 48 h post stimulation.*P < 0.05, **P < 0.01, and ***P < 0.001 vs. no treatment. LPS lipopolysaccharide
Mentions: Btk is involved in signal transduction by FcγRs expressed by myeloid lineage cells such as monocytes, macrophages, and neutrophils [15]. Therefore, we next examined the effect of HM71224 on inflammatory cytokine production by human monocytes. Human CD14+ cells were isolated from healthy volunteers and stimulated with heat-inactivated ICs in the presence of increasing concentrations of HM71224. The production of TNF-α, IL-1β, and IL-6 was inhibited in a dose-dependent manner (Fig. 4a). Btk also plays a crucial role in activating human PDCs via the TLR9 signaling pathway [16]. As expected, stimulation of human PDCs with either LPS (a TLR4 agonist) or CpG oligodeoxynucleotides (CpG ODN, a TLR9 agonist) induced the production of TNF-α and IL-6. HM71224 inhibited CpG-mediated cytokine production but not that mediated by LPS (Fig. 4b).Fig. 4

Bottom Line: The in vitro effects of HM71224 on B cells and monocytes were analyzed by examining phosphorylation of Btk and its downstream signaling molecules, along with cytokine production and osteoclast formation.Furthermore, HM71224 effectively inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β by human monocytes, and osteoclast formation by human monocytes.Thus, HM71224 is a potential novel therapeutic agent for rheumatoid arthritis in humans.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Department of Internal Medicine, Seoul National University College of Medicine, 101 Daehank-no Chongno-gu, Seoul, 03080, Korea.

ABSTRACT

Background: Bruton's tyrosine kinase (Btk) is critical for activation of B cells and myeloid cells. This study aimed to characterize the effects of HM71224, a novel Btk inhibitor, both in vitro and in a mouse model of experimental arthritis.

Methods: The kinase inhibition profile of HM71224 was analyzed. The in vitro effects of HM71224 on B cells and monocytes were analyzed by examining phosphorylation of Btk and its downstream signaling molecules, along with cytokine production and osteoclast formation. The in vivo effects of HM71224 were investigated in a mouse model of collagen-induced arthritis (CIA).

Results: HM71224 irreversibly bound to and inhibited Btk (IC50 = 1.95 nM). The compound also inhibited the phosphorylation of Btk and its downstream molecules such as PLCγ2, in activated Ramos B lymphoma cells and primary human B cells in a dose-dependent manner. Furthermore, HM71224 effectively inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β by human monocytes, and osteoclast formation by human monocytes. Finally, HM71224 improved experimental arthritis and prevented joint destruction in a murine model of CIA.

Conclusions: HM71224 inhibits Btk in B cells and monocytes and ameliorates experimental arthritis in a mouse model. Thus, HM71224 is a potential novel therapeutic agent for rheumatoid arthritis in humans.

No MeSH data available.


Related in: MedlinePlus