A synthetic compound, 4-acetyl-3-methyl-6-(3,4,5-trimethoxyphenyl)pyrano[3,4-c]pyran-1,8-dione, ameliorates ovalbumin-induced asthma

doi:10.1016/j.bmc.2013.08.045

Bioorganic & Medicinal Chemistry

Volume 21, Issue 21, 1 November 2013, Pages 6359-6365

https://doi.org/10.1016/j.bmc.2013.08.045 Get rights and content

Abstract

Eosinophilia is one of the characteristic signs of allergic inflammation. Massive migration of eosinophils to the airways can cause epithelial tissue injury, contraction of airway smooth muscle and increased bronchial responsiveness. Previously, we discovered a new compound, 1H,8H-pyrano[3,4-c]pyran-1,8-dione (PPY), derived from the fruit of Vitex rotundifolia L. and evaluated its anti-inflammatory and anti-asthmatic properties. In this study, we synthesized a new modified compound, 4-acetyl-3-methyl-6-(3,4,5-trimethoxyphenyl) pyrano[3,4-c]pyran-1,8-dione (PPY-345), which was based on the PPY skeleton, and we evaluated its anti-asthmatic effects. To evaluate the anti-asthmatic effect of PPY-345 in vitro, A549 lung epithelial cells were stimulated with TNF-alpha, IL-4 and IL-1-beta to induce the expression of CCL11 (Eotaxin), a chemokine involved in eosinophil chemotaxis. To characterize the anti-asthmatic properties of PPY-345 in vivo, we examined the influence of PPY-345 in an ovalbumin (OVA)-induced asthma model. PPY-345 treatments significantly reduced CCL11 secretion. PPY-345 treatment did not inhibit the translocation of NF-κB into the nucleus but suppressed the phosphorylation of signal transducers and activators of transcription 6 (STAT6). PPY-345 treatment significantly reduced airway hyperreactivity as measured by whole-body plethysmography. PPY-345 further reduced total cells, including eosinophil, macrophage and lymphocytes, in the BAL fluid, goblet cell hyperplasia and myosin light chain 2 positive smooth muscle cell area in the lung tissue. Additionally, PPY-345 significantly suppressed the levels of OVA–IgE present in the serum. These results suggested that PPY-345 could improve asthma symptoms in OVA-sensitized mice.

Graphical abstract

Introduction

Asthma is a chronic inflammatory disease characterized by airway inflammation, increased mucus production, intermittent airway obstruction and hyperresponsiveness.1, 2 Airway remodeling is specifically characterized by structural and morphometric changes to the airway, including subepithelial fibrosis, epithelial hypertrophy, goblet cell hyperplasia and smooth muscle hypertrophy.3, 4 The initial symptoms of asthma include airway inflammation, in which eosinophils play a crucial role.⁵ Eosinophils are always present in excess in the airways of asthma patients, but their accumulation in the lungs decreases as asthmatic symptoms decrease. CCL11 (Eotaxin) is a small protein that is produced in the lungs of asthmatic patients and is a potent eosinophil chemoattractant. CCL11, a CC chemokine, stimulates the migration of eosinophils from small blood vessels to lung tissue by acting on the CC chemokine receptor CCR3, located on the leukocyte cell surface.⁶ Asthmatic patients commonly take two types of medication: preventative and relief agents.⁷ Oral or inhaled steroids are common drugs for the treatment or prevention of asthma, but steroids have many unpleasant side effects. Thus, research is currently underway to find asthma treatment alternatives.⁸

We previously isolated a novel natural compound, 1H,8H-Pyrano[3,4-c]pyran-1,8-dione (PPY), from the fruit of Vitex rotundifolia L. and evaluated its anti-inflammatory and anti-asthmatic effects. We reported that PPY treatment significantly reduced the expression of eotaxin, IL-8, IL-16, and vascular cell adhesion molecule-1 (VCAM-1) mRNA in A549 lung epithelial cells that had been stimulated with cytokines. Additionally, in an ovalbumin-challenged mouse model, PPY profoundly inhibited eosinophil accumulation in the airways and reduced the levels of IL-4, IL-5, and IL-13 in the bronchoalveolar lavage fluid (BALF).⁹ As a follow-up study, we have synthesized a new compound, 4-acetyl-3-methyl-6-(3,4,5-trimethoxyphenyl)pyrano[3,4-c]pyran-1,8-dione (PPY-345) based on the PPY skeleton and evaluated its anti-asthmatic effects in vivo and in vitro.

Section snippets

Cells

A549 cells, a human type II-like epithelial lung cell line, were obtained from the Korean Cell Line Bank (Cancer Research Institute, Seoul, Korea). The cells were cultured in 100 mm tissue culture plates (Corning, Corning, NY, USA) in RPMI 1640 medium (Welgene, Daegu, Korea) supplemented with 10% heat-inactivated fetal bovine serum (Hyclone, Logan, UT, USA) and 100 U ml⁻¹ penicillin–streptomycin (Invitrogen, Rockville, MD, USA). The plates were incubated at 37 °C at 100% humidity and 5% CO₂. The

PPY-345 inhibits CCL11 secretion by cytokine-stimulated lung epithelial cells

To investigate the effects of PPY-345 on cultured cells, cell viability was monitored using the MTS assay in A549 lung epithelial cells. Concentrations of PPY-345 high as 50 μM did not affect cell viability (Fig. 3A). A solution of 0.5% ethanol was used as a vehicle control, which did not show any significant effects on A549 viability (data not shown). It is known that CCL11 (eotaxin-1) produced by lung epithelial cells is a potent chemoattractant for eosinophils. Therefore, inhibiting CCL11

Discussion

In our previous study, we screened the anti-asthmatic effects of 270 medicinal plants and we found that the fruit of Vitex rotundifolia L. had the highest effects in our in vitro experiments. After several rounds of fractionation and activity testing, a novel compound named PPY was isolated.⁹ As a follow-up study, we synthesized a PPY derivative based on the PPY skeleton and evaluated its activity. We demonstrated that PPY-345 inhibited the production of CCL11 in proinflammatory

Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea Government [MEST] (No. 2012-0005755).

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A synthetic compound, 4-acetyl-3-methyl-6-(3,4,5-trimethoxyphenyl)pyrano[3,4-c]pyran-1,8-dione, ameliorates ovalbumin-induced asthma

Abstract

Graphical abstract

Introduction

Section snippets

Cells

PPY-345 inhibits CCL11 secretion by cytokine-stimulated lung epithelial cells

Discussion

Acknowledgments

Lancet

Chest

J. Allergy Clin. Immunol.

Curr. Opin. Pharmacol.

Pediatr. Clin. North Am.

Tetrahedron Lett.

J. Allergy Clin. Immunol.

Immunity

Am. J. Respir. Crit. Care Med.

Am. Rev. Respir. Dis.