Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro

Elisangela Oliveira Freitas, Dirlei Nico, Rong Guan, José Roberto Meyer-Fernandes, Keith Clinch, Gary B. Evans, Peter C. Tyler, Vern L. Schramm, Clarisa B. Palatnik-de-Sousa

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Chemotherapy against visceral leishmaniasis is associated with high toxicity and drug resistance. Leishmania parasites are purine auxotrophs that obtain their purines from exogenous sources. Nucleoside hydrolases release purines from nucleosides and are drug targets for anti-leishmanial drugs, absent in mammal cells. We investigated the substrate specificity of the Leishmania (L.) donovani recombinant nucleoside hydrolase NH36 and the inhibitory effect of the immucillins IA (ImmA), DIA (DADMe-ImmA), DIH (DADMe-ImmH), SMIH (SerMe-ImmH), IH (ImmH), DIG (DADMe-ImmG), SMIG (SerMe-ImmG) and SMIA (SerME-ImmA) on its enzymatic activity. The inhibitory effects of immucillins on the in vitro multiplication of L. (L.) infantum chagasi and L. (L.) amazonensis promastigotes were determined using 0.05-500 μM and, when needed, 0.01-50 nM of each drug. The inhibition on multiplication of L. (L.) infantum chagasi intracellular amastigotes in vitro was assayed using 0.5, 1, 5 and 10 μM of IA, IH and SMIH. The NH36 shows specificity for inosine, guanosine, adenosine, uridine and cytidine with preference for adenosine and inosine. IA, IH, DIH, DIG, SMIH and SMIG immucillins inhibited L. (L.) infantum chagasi and L. (L.) amazonensis promastigote growth in vitro at nanomolar to micromolar concentrations. Promastigote replication was also inhibited in a chemically defined medium without a nucleoside source. Addition of adenosine decreases the immucillin toxicity. IA and IH inhibited the NH36 enzymatic activity (Ki = 0.080 μM for IA and 0.019 μM for IH). IA, IH and SMIH at 10 μM concentration, reduced the in vitro amastigote replication inside mice macrophages by 95% with no apparent effect on macrophage viability. Transmission electron microscopy revealed global alterations and swelling of L. (L.) infantum chagasi promastigotes after treatment with IA and IH while SMIH treatment determined intense cytoplasm vacuolization, enlarged vesicles and altered kinetoplasts. Our results suggest that IA, IH and SMIH may provide new chemotherapy agents for leishmaniasis.

Original languageEnglish (US)
Article numbere0124183
JournalPLoS One
Volume10
Issue number4
DOIs
StatePublished - Apr 24 2015

Fingerprint

Leishmania infantum
promastigotes
nucleosides
Leishmania
adenosine
N-Glycosyl Hydrolases
purines
Adenosine
amastigotes
Inosine
Chemotherapy
Macrophages
hydrolases
Pharmaceutical Preparations
drug therapy
Toxicity
macrophages
exogenous sources
toxicity
Purine Nucleosides

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Freitas, E. O., Nico, D., Guan, R., Meyer-Fernandes, J. R., Clinch, K., Evans, G. B., ... Palatnik-de-Sousa, C. B. (2015). Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro. PLoS One, 10(4), [e0124183]. https://doi.org/10.1371/journal.pone.0124183

Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro. / Freitas, Elisangela Oliveira; Nico, Dirlei; Guan, Rong; Meyer-Fernandes, José Roberto; Clinch, Keith; Evans, Gary B.; Tyler, Peter C.; Schramm, Vern L.; Palatnik-de-Sousa, Clarisa B.

In: PLoS One, Vol. 10, No. 4, e0124183, 24.04.2015.

Research output: Contribution to journalArticle

Freitas, EO, Nico, D, Guan, R, Meyer-Fernandes, JR, Clinch, K, Evans, GB, Tyler, PC, Schramm, VL & Palatnik-de-Sousa, CB 2015, 'Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro', PLoS One, vol. 10, no. 4, e0124183. https://doi.org/10.1371/journal.pone.0124183
Freitas EO, Nico D, Guan R, Meyer-Fernandes JR, Clinch K, Evans GB et al. Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro. PLoS One. 2015 Apr 24;10(4). e0124183. https://doi.org/10.1371/journal.pone.0124183
Freitas, Elisangela Oliveira ; Nico, Dirlei ; Guan, Rong ; Meyer-Fernandes, José Roberto ; Clinch, Keith ; Evans, Gary B. ; Tyler, Peter C. ; Schramm, Vern L. ; Palatnik-de-Sousa, Clarisa B. / Immucillins impair Leishmania (L.) infantum chagasi and Leishmania (L.) amazonensis multiplication In Vitro. In: PLoS One. 2015 ; Vol. 10, No. 4.
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abstract = "Chemotherapy against visceral leishmaniasis is associated with high toxicity and drug resistance. Leishmania parasites are purine auxotrophs that obtain their purines from exogenous sources. Nucleoside hydrolases release purines from nucleosides and are drug targets for anti-leishmanial drugs, absent in mammal cells. We investigated the substrate specificity of the Leishmania (L.) donovani recombinant nucleoside hydrolase NH36 and the inhibitory effect of the immucillins IA (ImmA), DIA (DADMe-ImmA), DIH (DADMe-ImmH), SMIH (SerMe-ImmH), IH (ImmH), DIG (DADMe-ImmG), SMIG (SerMe-ImmG) and SMIA (SerME-ImmA) on its enzymatic activity. The inhibitory effects of immucillins on the in vitro multiplication of L. (L.) infantum chagasi and L. (L.) amazonensis promastigotes were determined using 0.05-500 μM and, when needed, 0.01-50 nM of each drug. The inhibition on multiplication of L. (L.) infantum chagasi intracellular amastigotes in vitro was assayed using 0.5, 1, 5 and 10 μM of IA, IH and SMIH. The NH36 shows specificity for inosine, guanosine, adenosine, uridine and cytidine with preference for adenosine and inosine. IA, IH, DIH, DIG, SMIH and SMIG immucillins inhibited L. (L.) infantum chagasi and L. (L.) amazonensis promastigote growth in vitro at nanomolar to micromolar concentrations. Promastigote replication was also inhibited in a chemically defined medium without a nucleoside source. Addition of adenosine decreases the immucillin toxicity. IA and IH inhibited the NH36 enzymatic activity (Ki = 0.080 μM for IA and 0.019 μM for IH). IA, IH and SMIH at 10 μM concentration, reduced the in vitro amastigote replication inside mice macrophages by 95{\%} with no apparent effect on macrophage viability. Transmission electron microscopy revealed global alterations and swelling of L. (L.) infantum chagasi promastigotes after treatment with IA and IH while SMIH treatment determined intense cytoplasm vacuolization, enlarged vesicles and altered kinetoplasts. Our results suggest that IA, IH and SMIH may provide new chemotherapy agents for leishmaniasis.",
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