LCB 03-0110, a Novel Pan-Discoidin Domain Receptor/c-Src Family Tyrosine Kinase Inhibitor, Suppresses Scar Formation by Inhibiting Fibroblast and Macrophage Activation

Abstract

Wound healing generally induces an inflammatory response associated with tissue fibrosis in which activated macrophage and myofibroblast cells are primarily involved. Although this is known to be the underlying mechanism for scarring and various fibrotic pathologies, no effective intervention is currently available. We identified (3-(2-(3-(morpholinomethyl)phenyl)thieno[3,2-b]pyridin-7-ylamino)phenol (LCB 03-0110), a thienopyridine derivative, as a potent inhibitor of discoidin domain receptor family tyrosine kinases and discovered that this compound strongly inhibits several tyrosine kinases, including the c-Src family, spleen tyrosine kinase, Bruton's tyrosine kinase, and vascular endothelial growth factor receptor 2, which are important for immune cell signaling and inflammatory reactions. LCB 03-0110 suppressed the proliferation and migration of primary dermal fibroblasts induced by transforming growth factor ␤1 and type I collagen, and this result correlated with the inhibition ability of the compound against enhanced expression of ␣-smooth muscle actin and activation of Akt1 and focal adhesion kinase. In J774A.1 macrophage cells activated by lipopolysaccharide LCB 03-0110 inhibited cell migration and nitric oxide, inducible nitric-oxide synthase, cyclooxygenase 2, and tumor necrosis factor-␣ synthesis. LCB 03-0110 applied topically to full excisional wounds on rabbit ears suppressed the accumulation of myofibroblast and macrophage cells in the healing wound and reduced hypertrophic scar formation after wound closing, without delaying the wound closing process. Taken together, the pharmacological activities of LCB 03-0110 suggest that it could be an effective agent for suppressing fibroinflammation by simultaneously targeting activated fibroblasts and macrophages.

Quercetin Shows the Pharmacological Activity to Simultaneously Downregulate the Inflammatory and Fibrotic Responses to Tissue Injury in Association with its Ability to Target Multi-Kinases

“…In addition to macrophages, we also showed that quercetin inhibits the cell signaling for activating dermal fibroblasts in responding to treatment with collagen and TGF-β1. It was reported that TGF-β1 is a key cytokine that induces dermal fibroblasts to undergo the activation into myofibroblast in cutaneous scarring [54] and that collagen synergized with TGF-β1 for this activation [35]. Our results from these cell culture experiments were consistent with the outcome of our animal model experiment in this study, which demonstrated that quercetin treatment reduces the number of activated macrophages and myofibroblasts at the cutaneous injury site of rabbit ear.…”

“…We activated primary dermal fibroblasts by plating them on type I collagen coated wells and culturing in a medium containing 5 ng/mL TGF-β1 and 0.5% FBS. In a previous study, this culture condition induces the activating phosphorylations in the signaling molecules of Akt1 and FAK and subsequently enhances the expression of α-SMA and the migratory ability of the cells, which are typical biomarkers of activation into myofibroblast-type cells [35]. After an hour with this culture condition, we observed an increase in the activating phosphorylation of tyrosine residues 397 and 861 of FAK and serine 473 of Akt1 without any effect on their total amount (Fig.…”

“…For activation of primary dermal fibroblasts, the cells were plated in 6-well plate coated with 200 μg/mL type 1 collagen (Sigma) using DMEM containing 0.5% FBS and 5 ng/mL of TGF-β1 (R&D Systems). The auto-tyrosine phosphorylation assay of cellular DDR2 was performed using human embryonic kidney (HEK) 293 cells stably transfected with human DDR2 by treating with collagen type 1 as described previously [35].…”

Discoidin domain receptor 1 (DDR1) kinase as target for structure-based drug discovery

“…Sun et al . identified (3-(2-(3-(morpholinomethyl)phenyl)thieno [3,2-b]pyridin-7-ylamino) phenol (LCB 03-0110 in Figure 3) as a potent inhibitor of both DDR1 and DDR2 along with several other tyrosine kinases [29]. Recently, Gao et al .…”

“…Flexible docking with ROSETTALIGAND into crystal structures would improve docking accuracy. In addition, as the number of known DDR1 inhibitors grows [25–27,29–31,38], ligand-based drug discovery methods, such as quantitative structure–activity relation (SAR) models, could be developed. The discovery of inhibitors of DDR1 kinase activity might be beneficial in several pathological conditions where DDR1 has been identified as a potential therapeutic target.…”