Designing Anti-inflammatory Drugs from Parasitic Worms: A Synthetic Small Molecule Analogue of the Acanthocheilonema viteae Product ES-62 Prevents Development of Collagen-Induced Arthritis

Abstract

In spite of increasing evidence that parasitic worms may protect humans from developing allergic and autoimmune diseases and the continuing identification of defined helminth-derived immunomodulatory molecules, to date no new anti-inflammatory drugs have been developed from these organisms. We have approached this matter in a novel manner by synthesizing a library of drug-like small molecules based upon phosphorylcholine, the active moiety of the anti-inflammatory Acanthocheilonema viteae product, ES-62, which as an immunogenic protein is unsuitable for use as a drug. Following preliminary in vitro screening for inhibitory effects on relevant macrophage cytokine responses, a sulfone-containing phosphorylcholine analogue (11a) was selected for testing in an in vivo model of inflammation, collagen-induced arthritis (CIA). Testing revealed that 11a was as effective as ES-62 in protecting DBA/1 mice from developing CIA and mirrored its mechanism of action in downregulating the TLR/IL-1R transducer, MyD88. 11a is thus a novel prototype for anti-inflammatory drug development.

The helminth product, ES-62 modulates dendritic cell responses by inducing the selective autophagolysosomal degradation of TLR-transducers, as exemplified by PKCδ

“…On day 6, immature bmDCs (65–85% CD11c + ) were harvested by gentle scraping and these bmDCs were further purified (≥95%) by positive selection using CD11c-MACS microbead separation (Miltenyi Biotec, Woking, UK) according to the manufacturer’s instructions. CD11c + DCs (2 × 10 6 cells) were then incubated in medium ± ES-62 (2 μg/ml) and/or lipopolysaccharide (LPS; 1 μg/ml) ( Salmonella minnesota or, where indicated, Escherichia coli , strain 055:B5 [Sigma, Poole UK]) prior to detection of cytokine release by ELISA and/or cell signalling events by Western Blotting7817225758.…”

“…For bmDC-T cell co-cultures, DCs were incubated in medium ± ES-62 and/or LPS, prior to pulsing with OVA 323–339 peptide (0–300 nM) before incubation with naive OVA-specific CD4 + CD62L + T cells from DO.11.10.BALB/c or OT-II.C57BL/6 mice, isolated using Miltenyi magnetic bead technology421545558. In some experiments, the transgenic (tg) TCR T cells, detected using the clonotypic monoclonal Ab KJ1.26, were labelled with 5, 6-Carboxy-Succinimidyl-Fluorescein-Ester (CFSE; 5 μM) and assessed for cell phenotype and proliferation by flow cytometry as previously described, using a Becton Dickinson FACSCalibar TM flow cytometer (BD Biosciences, Oxford UK) and analysed using Flowjo software (Tree Star Inc, OR, USA, version 8.8.6)452154.…”

From the worm to the pill, the parasitic worm product ES-62 raises new horizons in the treatment of rheumatoid arthritis

“…Confirming this, 11a was shown to be as effective as ES-62 in protecting mice against CIA, via downregulation of pathogenic IL-17/IFNg responses. 71 …”

Helminth Infections Decrease Host Susceptibility to Immune-Mediated Diseases

“…ES-62 also works directly on polarized Th17 cells to reduce IL17 production and Myd88 expression (87). A synthetic small molecule modeled after ES-62 (N-(2-[(4-bromobenzyl)sulfonyl]ethyl)-N,N-dimethylamine, 11a) inhibits development of CIA and suppresses release of IL12p40 and IL6 from LPS-stimulated macrophages (88). …”