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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.

Al-Riyami L, Pineda MA, Rzepecka J, Huggan JK, Khalaf AI, Suckling CJ, Scott FJ, Rodgers DT, Harnett MM, Harnett W - J. Med. Chem. (2013)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde , 161 Cathedral Street, Glasgow G4 0RE, U.K.

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.

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11a inhibitsTh17 polarization. (A) Serum IL-17 levelsare plotted as mean values of triplicate IL-17 analyses from individualmice (PBS, n = 12; 11a, n = 13). (B) BmDCs from C57BL/6 mice were preincubated with or without(11a) (5 μg/mL) for 2 h prior to stimulation withLPS for 24 h, and TNFα, IL-6, and IL-23 levels were then analyzed.Data are the mean values (of triplicate samples) ± SEM pooledfrom 4 independent experiments. (C) BmDCs from BALB/c mice preincubatedwith or without 11a (5 μg/mL) were pulsed withthe indicated concentration of OVA peptide and cocultured with naiveOVA-specific CD4+ T cells (DO.11.10/BALB/c) for 3 daysbefore measuring IL-17 release. Data are the mean values ± SDof duplicate samples pooled from two independent experiments (n = 4). (D) Pooled normalized data from four independentexperiments analyzing the effect of 11a on IL-17 releasefrom bmDC (C57BL/6)-OVA-specific CD4+T cell (OT-II/C57BL/6)cocultures. Data are presented as the means of the mean percentagemaximum (LPS) response ± SEM where data were normalized to theLPS response at 10 nM (left), 100 nM (middle), and 300 nM (right)OVA, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001.
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fig7: 11a inhibitsTh17 polarization. (A) Serum IL-17 levelsare plotted as mean values of triplicate IL-17 analyses from individualmice (PBS, n = 12; 11a, n = 13). (B) BmDCs from C57BL/6 mice were preincubated with or without(11a) (5 μg/mL) for 2 h prior to stimulation withLPS for 24 h, and TNFα, IL-6, and IL-23 levels were then analyzed.Data are the mean values (of triplicate samples) ± SEM pooledfrom 4 independent experiments. (C) BmDCs from BALB/c mice preincubatedwith or without 11a (5 μg/mL) were pulsed withthe indicated concentration of OVA peptide and cocultured with naiveOVA-specific CD4+ T cells (DO.11.10/BALB/c) for 3 daysbefore measuring IL-17 release. Data are the mean values ± SDof duplicate samples pooled from two independent experiments (n = 4). (D) Pooled normalized data from four independentexperiments analyzing the effect of 11a on IL-17 releasefrom bmDC (C57BL/6)-OVA-specific CD4+T cell (OT-II/C57BL/6)cocultures. Data are presented as the means of the mean percentagemaximum (LPS) response ± SEM where data were normalized to theLPS response at 10 nM (left), 100 nM (middle), and 300 nM (right)OVA, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001.

Mentions: Unlike in our earlier study with ES-62,911a did not induce a statistically significantreductionin the serum levels of IL-17 in these mice (Figure 7A). However, in our previous study,9 it was shown that pre-exposure to ES-62 inhibited the capacity ofdendritic cells (bmDCs) to respond to LPS by secretion of the pro-inflammatorycytokine TNF-α and two cytokines, IL-6 and IL-23, associatedwith polarization and maintenance of Th17 cells, respectively, andwhen these experiments were repeated with the bmDCs pre-exposed to 11a, production of all three cytokines was significantly reduced(Figure 7B). As with ES-62,4 this did not reflect downregulation of TLR4 expressionor reduction in bmDC viability (data not shown). Furthermore, consistentwith the ability of 11a to inhibit production of thetwo Th17-promoting cytokines, 11a-treated DCs demonstratea reduced ability to drive naïve antigen (OVA)-specificTh cells toward a Th17 phenotype, as evidenced by suppression of OVA-specificIL-17 production in such bmDC-CD4+ T cell cocultures (Figure 7C,D). Again, this is consistent with what was previouslyobserved with ES-62.9


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.

Al-Riyami L, Pineda MA, Rzepecka J, Huggan JK, Khalaf AI, Suckling CJ, Scott FJ, Rodgers DT, Harnett MM, Harnett W - J. Med. Chem. (2013)

11a inhibitsTh17 polarization. (A) Serum IL-17 levelsare plotted as mean values of triplicate IL-17 analyses from individualmice (PBS, n = 12; 11a, n = 13). (B) BmDCs from C57BL/6 mice were preincubated with or without(11a) (5 μg/mL) for 2 h prior to stimulation withLPS for 24 h, and TNFα, IL-6, and IL-23 levels were then analyzed.Data are the mean values (of triplicate samples) ± SEM pooledfrom 4 independent experiments. (C) BmDCs from BALB/c mice preincubatedwith or without 11a (5 μg/mL) were pulsed withthe indicated concentration of OVA peptide and cocultured with naiveOVA-specific CD4+ T cells (DO.11.10/BALB/c) for 3 daysbefore measuring IL-17 release. Data are the mean values ± SDof duplicate samples pooled from two independent experiments (n = 4). (D) Pooled normalized data from four independentexperiments analyzing the effect of 11a on IL-17 releasefrom bmDC (C57BL/6)-OVA-specific CD4+T cell (OT-II/C57BL/6)cocultures. Data are presented as the means of the mean percentagemaximum (LPS) response ± SEM where data were normalized to theLPS response at 10 nM (left), 100 nM (middle), and 300 nM (right)OVA, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001.
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fig7: 11a inhibitsTh17 polarization. (A) Serum IL-17 levelsare plotted as mean values of triplicate IL-17 analyses from individualmice (PBS, n = 12; 11a, n = 13). (B) BmDCs from C57BL/6 mice were preincubated with or without(11a) (5 μg/mL) for 2 h prior to stimulation withLPS for 24 h, and TNFα, IL-6, and IL-23 levels were then analyzed.Data are the mean values (of triplicate samples) ± SEM pooledfrom 4 independent experiments. (C) BmDCs from BALB/c mice preincubatedwith or without 11a (5 μg/mL) were pulsed withthe indicated concentration of OVA peptide and cocultured with naiveOVA-specific CD4+ T cells (DO.11.10/BALB/c) for 3 daysbefore measuring IL-17 release. Data are the mean values ± SDof duplicate samples pooled from two independent experiments (n = 4). (D) Pooled normalized data from four independentexperiments analyzing the effect of 11a on IL-17 releasefrom bmDC (C57BL/6)-OVA-specific CD4+T cell (OT-II/C57BL/6)cocultures. Data are presented as the means of the mean percentagemaximum (LPS) response ± SEM where data were normalized to theLPS response at 10 nM (left), 100 nM (middle), and 300 nM (right)OVA, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001.
Mentions: Unlike in our earlier study with ES-62,911a did not induce a statistically significantreductionin the serum levels of IL-17 in these mice (Figure 7A). However, in our previous study,9 it was shown that pre-exposure to ES-62 inhibited the capacity ofdendritic cells (bmDCs) to respond to LPS by secretion of the pro-inflammatorycytokine TNF-α and two cytokines, IL-6 and IL-23, associatedwith polarization and maintenance of Th17 cells, respectively, andwhen these experiments were repeated with the bmDCs pre-exposed to 11a, production of all three cytokines was significantly reduced(Figure 7B). As with ES-62,4 this did not reflect downregulation of TLR4 expressionor reduction in bmDC viability (data not shown). Furthermore, consistentwith the ability of 11a to inhibit production of thetwo Th17-promoting cytokines, 11a-treated DCs demonstratea reduced ability to drive naïve antigen (OVA)-specificTh cells toward a Th17 phenotype, as evidenced by suppression of OVA-specificIL-17 production in such bmDC-CD4+ T cell cocultures (Figure 7C,D). Again, this is consistent with what was previouslyobserved with ES-62.9

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde , 161 Cathedral Street, Glasgow G4 0RE, U.K.

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.

Show MeSH
Related in: MedlinePlus