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Glycotriazole-peptides derived from the peptide HSP1: synergistic effect of triazole and saccharide rings on the antifungal activity

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Abstract

This work proposes a strategy that uses solid-phase peptide synthesis associated with copper(I)-catalyzed azide alkyne cycloaddition reaction to promote the glycosylation of an antimicrobial peptide (HSP1) containing a carboxyamidated C-terminus (HSP1-NH2). Two glycotriazole-peptides, namely [p-Glc-trz-G1]HSP1-NH2 and [p-GlcNAc-trz-G1]HSP1-NH2, were prepared using per-O-acetylated azide derivatives of glucose and N-acetylglucosamine in the presence of copper(II) sulfate pentahydrate (CuSO4·5H2O) and sodium ascorbate as a reducing agent. In order to investigate the synergistic action of the carbohydrate motif linked to the triazole-peptide structure, a triazole derivative [trz-G1]HSP1-NH2 was also prepared. A set of biophysical approaches such as DLS, Zeta Potential, SPR and carboxyfluorescein leakage from phospholipid vesicles confirmed higher membrane disruption and lytic activities as well as stronger peptide-LUVs interactions for the glycotriazole-peptides when compared to HSP1-NH2 and to its triazole derivative, which is in accordance with the performed biological assays: whereas HSP1-NH2 presents relatively low and [trz-G1]HSP1-NH2 just moderate fungicidal activity, the glycotriazole-peptides are significantly more effective antifungal agents. In addition, the glycotriazole-peptides and the triazole derivative present strong inhibition effects on ergosterol biosynthesis in Candida albicans, when compared to HSP1-NH2 alone. In conclusion, the increased fungicidal activity of the glycotriazole-peptides seems to be the result of (A) more pronounced membrane-disruptive properties, which is related to the presence of a saccharide ring, together with (B) the inhibition of ergosterol biosynthesis, which seems to be related to the presence of both the monosaccharide and the triazole rings.

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Abbreviations

AMPs:

Antimicrobial peptides

CD:

Circular dichroism spectroscopy

CF:

Carboxyfluorescein

CF%:

Percentage of carboxyfluorescein release

CuAAC:

Copper(I)-catalyzed azide-alkyne cycloaddition

DCM:

Dichloromethane

D h :

Hydrodynamic diameter

DIC:

Diisopropylcarbodiimide

DLS:

Dynamic light scattering

DMF:

N,N-dimethylformamide

EDTA:

Ethylenediaminetetraacetic acid

HOBt:

1-Hydroxybenzotriazole

IPA:

Isopropyl alcohol

LUVs:

Large unilamellar vesicles

MIC:

Minimum inhibitory concentration

PIPE:

4-Methylpiperidine

POPC:

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

POPG:

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol

SPR:

Surface plasmon resonance

Tris:

2-Amino-2-hydroxymethyl-propane-1,3-diol

HSP1:

Hylaseptin-P1

HSP1-NH2 :

Carboxyamidated C-terminus hylaseptin-P1

[Pra1]HSP1-NH2 :

Propargylated HSP1-NH2

[p-GlcNAc-trz-G1]HSP1-NH2 :

per-O-acetylated N-acetilglucosamine-triazole derived from HSP1-NH2

[p-Glc-trz-G1]HSP1-NH2 :

per-O-acetylated N-acetilglucose-triazole derived from HSP1-NH2

[trz-G1]HSP1-NH2 :

Triazole-peptide derived from HSP1-NH2

THF:

Tetrahydrofuran

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Acknowledgements

EFCJ, FCLA and LLP acknowledge Grants from UFVJM and CNPq. CFRCG, JDSF, LLF and RJA acknowledge Grants from CNPq. This work is a collaboration research project of members of the Rede Mineira de Química (RQ-MG) supported by FAPEMIG (Project: CEX-RED-00010-14). We also acknowledge financial support from FAPEMIG, CAPES, PRPq-UFMG and PRPPG-UFVJM.

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Author notes

  1. Marcelo P. Bemquerer

    Present address: Laboratório de Espectrometria de Massa, Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-Recursos Genéticos e Biotecnologia, Estação Parque Biológico, Final W5, Asa Norte, Brasília, DF, 70770-900, Brazil

  2. Eduardo F. C. Junior and Carlos F. R. C. Guimarães contributed equally to this work. The manuscript was written through contributions of all authors.

Authors and Affiliations

  1. Departamento de Química, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, MG, 39100-000, Brazil

    Eduardo F. C. Junior & Rodrigo M. Verly

  2. Departamento de Química, Universidade Federal de Minas Gerais, PO Box 486, Belo Horizonte, MG, 31270-901, Brazil

    Carlos F. R. C. Guimarães, José D. de Souza Filho & Jarbas M. Resende

  3. Faculdade de Farmácia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, MG, 39100-000, Brazil

    Kelly C. Kato & Helen R. Martins

  4. Faculdade de Farmácia, Universidade Federal de Minas Gerais, PO Box 486, Belo Horizonte, MG, 31270-901, Brazil

    Lucas L. Franco & Ricardo J. Alves

  5. Instituto de Ciência e Tecnologia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, MG, 39100-000, Brazil

    Victor H. O. Munhoz

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Correspondence to Rodrigo M. Verly.

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The authors declare that they have no conflict of interest.

Funding

This work was sponsored by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM). This work is a collaboration research project of members of the Rede Mineira de Química (RQ-MG) supported by FAPEMIG (Project: CEX-RED-00010-14) and of the Toxinology Network sponsored by CAPES (Process 230.38.006280/2011-07). CAPES offered a Grant to EFCJ (Master Grant). CNPq offered PhD Grants to CFRCG and LLF and researcher Grants to JDSF and RJA.

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Junior, E.F.C., Guimarães, C.F.R.C., Franco, L.L. et al. Glycotriazole-peptides derived from the peptide HSP1: synergistic effect of triazole and saccharide rings on the antifungal activity. Amino Acids 49, 1389–1400 (2017). https://doi.org/10.1007/s00726-017-2441-2

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