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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-related changes in macrophage cytokine productionand signaling of p65 NFκB in response to LPS, BLP, and CpG.BmMs were preincubated with 11a at 5 μg/mL for18 h prior to stimulation with 100 ng/mL LPS, 10 ng/mL BLP, or 0.01μM CpG for 24 h and analysis of levels of IL-12p40 (A) and IL-6(B) performed by ELISA. (C) Stimulation with PAMPs as above for LPSand BLP, and 1 μM CpG, but for 1 h and the level of p65 activationin duplicate samples measured by TransAM. For statistical analysisfor (A) and (B), *p < 0.05.
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fig3: 11a-related changes in macrophage cytokine productionand signaling of p65 NFκB in response to LPS, BLP, and CpG.BmMs were preincubated with 11a at 5 μg/mL for18 h prior to stimulation with 100 ng/mL LPS, 10 ng/mL BLP, or 0.01μM CpG for 24 h and analysis of levels of IL-12p40 (A) and IL-6(B) performed by ELISA. (C) Stimulation with PAMPs as above for LPSand BLP, and 1 μM CpG, but for 1 h and the level of p65 activationin duplicate samples measured by TransAM. For statistical analysisfor (A) and (B), *p < 0.05.

Mentions: Of directrelevance to the aims of the project, a number of the compounds reducedproduction of cytokines important for promoting Th1 and/or Th17 responses,IL-12p40 and/or IL-6, in response to one or more of the TLR ligands.However, some of these did not target either TLR4 (sulfonamide 21o) or TLR2 (sulfonamides 21n, 21o) responses. Likewise, of the carboxamides, 24c onlyeffectively targeted both CpG responses. The most promising compoundswere those that showed the broadest response to the PAMPs, the sulfones, 11a and 12b, the sulfonamide 21l, and the carboxamide 24b. 11a (see Figure 3A) and 12b both mimicked ES-62 in targetingIL-12p40 production via each of TLR2, 4, and 9, and this cytokineis pathogenic in CIA41 due to it beinga component of both IL-12p70 and IL-23, which promote Th1 and Th17responses, respectively, and hence a therapeutic target (ustekinumab)in inflammatory autoimmune diseases.42,43 By contrast,sulfonamide 21l and carboxamide 24b didnot target LPS- and BLP-mediated IL-12p40 responses, respectively.Sulfone 11a additionally targeted IL-6 production (seealso Figure 3B) in response to all three TLRs(although this did not reach statistical significance for TLR2 inall experiments), and this cytokine has also been shown to be pathogenic44 and thus a therapeutic target (tocilizumab)in RA.45 Although sulfone 12b could suppress CpG-mediated IL-6 responses, it was less effectiveat inhibiting such TLR2- or TLR4-coupled responses (decreases notreaching statistical significance), which have been shown to be importantin the development of inflammatory Th17 responses, including thosein arthritis.46−48 Moreover, as observed with ES-62,40,49 (11a (Figure 3C), but not 21l (results not shown), was able to inhibit TLR-mediatedp65 NF-κB activation in response to all three PAMPs.


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-related changes in macrophage cytokine productionand signaling of p65 NFκB in response to LPS, BLP, and CpG.BmMs were preincubated with 11a at 5 μg/mL for18 h prior to stimulation with 100 ng/mL LPS, 10 ng/mL BLP, or 0.01μM CpG for 24 h and analysis of levels of IL-12p40 (A) and IL-6(B) performed by ELISA. (C) Stimulation with PAMPs as above for LPSand BLP, and 1 μM CpG, but for 1 h and the level of p65 activationin duplicate samples measured by TransAM. For statistical analysisfor (A) and (B), *p < 0.05.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4125414&req=5

fig3: 11a-related changes in macrophage cytokine productionand signaling of p65 NFκB in response to LPS, BLP, and CpG.BmMs were preincubated with 11a at 5 μg/mL for18 h prior to stimulation with 100 ng/mL LPS, 10 ng/mL BLP, or 0.01μM CpG for 24 h and analysis of levels of IL-12p40 (A) and IL-6(B) performed by ELISA. (C) Stimulation with PAMPs as above for LPSand BLP, and 1 μM CpG, but for 1 h and the level of p65 activationin duplicate samples measured by TransAM. For statistical analysisfor (A) and (B), *p < 0.05.
Mentions: Of directrelevance to the aims of the project, a number of the compounds reducedproduction of cytokines important for promoting Th1 and/or Th17 responses,IL-12p40 and/or IL-6, in response to one or more of the TLR ligands.However, some of these did not target either TLR4 (sulfonamide 21o) or TLR2 (sulfonamides 21n, 21o) responses. Likewise, of the carboxamides, 24c onlyeffectively targeted both CpG responses. The most promising compoundswere those that showed the broadest response to the PAMPs, the sulfones, 11a and 12b, the sulfonamide 21l, and the carboxamide 24b. 11a (see Figure 3A) and 12b both mimicked ES-62 in targetingIL-12p40 production via each of TLR2, 4, and 9, and this cytokineis pathogenic in CIA41 due to it beinga component of both IL-12p70 and IL-23, which promote Th1 and Th17responses, respectively, and hence a therapeutic target (ustekinumab)in inflammatory autoimmune diseases.42,43 By contrast,sulfonamide 21l and carboxamide 24b didnot target LPS- and BLP-mediated IL-12p40 responses, respectively.Sulfone 11a additionally targeted IL-6 production (seealso Figure 3B) in response to all three TLRs(although this did not reach statistical significance for TLR2 inall experiments), and this cytokine has also been shown to be pathogenic44 and thus a therapeutic target (tocilizumab)in RA.45 Although sulfone 12b could suppress CpG-mediated IL-6 responses, it was less effectiveat inhibiting such TLR2- or TLR4-coupled responses (decreases notreaching statistical significance), which have been shown to be importantin the development of inflammatory Th17 responses, including thosein arthritis.46−48 Moreover, as observed with ES-62,40,49 (11a (Figure 3C), but not 21l (results not shown), was able to inhibit TLR-mediatedp65 NF-κB activation in response to all three PAMPs.

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