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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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Lack of toxic effect of 11a on macrophages.BmMs inultralow binding tissue culture plates were rested in RPMI completemedium for 24 h before culturing with fresh medium or medium containing 11a (5 μg/mL) or ES-62 (2 μg/mL). After 18 h,the macrophages were stimulated with medium containing 100 ng/mL LPSor 10 ng/mL BLP or 0.01 μM CpG for an additional 24 h beforestaining with 7-AAD to assess their viability. The samples were analyzedby flow cytometry, and the data are presented as density plots withfrequency of 7-AAD positive (dead) cells indicated in the gates. Theresults shown are from a single experiment representative of two independentexperiments.
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fig4: Lack of toxic effect of 11a on macrophages.BmMs inultralow binding tissue culture plates were rested in RPMI completemedium for 24 h before culturing with fresh medium or medium containing 11a (5 μg/mL) or ES-62 (2 μg/mL). After 18 h,the macrophages were stimulated with medium containing 100 ng/mL LPSor 10 ng/mL BLP or 0.01 μM CpG for an additional 24 h beforestaining with 7-AAD to assess their viability. The samples were analyzedby flow cytometry, and the data are presented as density plots withfrequency of 7-AAD positive (dead) cells indicated in the gates. Theresults shown are from a single experiment representative of two independentexperiments.

Mentions: Thus, as both IL-6 and IL-12p40 (via IL-23) promote the differentiationand maintenance of Th17 responses, it was considered that 11a was most likely to mimic the protective effects of ES-62 in CIAby suppressing production of IL-17,9 acytokine that is pathogenic in RA and an emerging therapeutic target(for example via the humanized monoclonal antibody LY2439821 and thehuman monoclonal antibody AIN457; reviews refs (42,50−52)). Limited analysis ofpotency revealed that 11a was still able to induce astatistically significant reduction of LPS-stimulated production ofIL-6 by macrophages when the concentration was reduced to 1 μg/mL,but significant effects were not consistently observed at 0.2 μg/mL(data not shown). Like ES-62, 11a showed no evidenceof toxicity under conditions mimicking the macrophage screen of cytokineproduction as determined using the cell viability indicator, 7-actinomycinD (7-AAD) (Figure 4). Indeed, the SMA showedsome evidence of protecting against the loss of cell viability associatedwith exposure to LPS. 11a was thus considered suitablefor testing in vivo.


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)

Lack of toxic effect of 11a on macrophages.BmMs inultralow binding tissue culture plates were rested in RPMI completemedium for 24 h before culturing with fresh medium or medium containing 11a (5 μg/mL) or ES-62 (2 μg/mL). After 18 h,the macrophages were stimulated with medium containing 100 ng/mL LPSor 10 ng/mL BLP or 0.01 μM CpG for an additional 24 h beforestaining with 7-AAD to assess their viability. The samples were analyzedby flow cytometry, and the data are presented as density plots withfrequency of 7-AAD positive (dead) cells indicated in the gates. Theresults shown are from a single experiment representative of two independentexperiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125414&req=5

fig4: Lack of toxic effect of 11a on macrophages.BmMs inultralow binding tissue culture plates were rested in RPMI completemedium for 24 h before culturing with fresh medium or medium containing 11a (5 μg/mL) or ES-62 (2 μg/mL). After 18 h,the macrophages were stimulated with medium containing 100 ng/mL LPSor 10 ng/mL BLP or 0.01 μM CpG for an additional 24 h beforestaining with 7-AAD to assess their viability. The samples were analyzedby flow cytometry, and the data are presented as density plots withfrequency of 7-AAD positive (dead) cells indicated in the gates. Theresults shown are from a single experiment representative of two independentexperiments.
Mentions: Thus, as both IL-6 and IL-12p40 (via IL-23) promote the differentiationand maintenance of Th17 responses, it was considered that 11a was most likely to mimic the protective effects of ES-62 in CIAby suppressing production of IL-17,9 acytokine that is pathogenic in RA and an emerging therapeutic target(for example via the humanized monoclonal antibody LY2439821 and thehuman monoclonal antibody AIN457; reviews refs (42,50−52)). Limited analysis ofpotency revealed that 11a was still able to induce astatistically significant reduction of LPS-stimulated production ofIL-6 by macrophages when the concentration was reduced to 1 μg/mL,but significant effects were not consistently observed at 0.2 μg/mL(data not shown). Like ES-62, 11a showed no evidenceof toxicity under conditions mimicking the macrophage screen of cytokineproduction as determined using the cell viability indicator, 7-actinomycinD (7-AAD) (Figure 4). Indeed, the SMA showedsome evidence of protecting against the loss of cell viability associatedwith exposure to LPS. 11a was thus considered suitablefor testing in vivo.

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