Synergistic antifungal interaction of N-(butylcarbamothioyl) benzamide and amphotericin B against Cryptococcus neoformans

Gabriella Maria Andriani; Lais Fernanda de Almeida Spoladori; Marciéli Fabris; Priscila Goes Camargo; Patrícia Morais Lopes Pereira; Jussevania Pereira Santos; Guilherme Bartolomeu-Gonçalves; Lais Alonso; Cesar Armando Contreras Lancheros; Antonio Alonso; Celso Vataru Nakamura; Fernando Macedo Jr; Phileno Pinge-Filho; Lucy Megumi Yamauchi; Marcelle de Lima Ferreira Bispo; Eliandro Reis Tavares; Sueli Fumie Yamada-Ogatta

doi:10.3389/fmicb.2023.1040671

Synergistic antifungal interaction of N-(butylcarbamothioyl) benzamide and amphotericin B against Cryptococcus neoformans

Front Microbiol. 2023 Mar 7:14:1040671. doi: 10.3389/fmicb.2023.1040671. eCollection 2023.

Authors

Gabriella Maria Andriani^{1

2}, Lais Fernanda de Almeida Spoladori^{1

2}, Marciéli Fabris³, Priscila Goes Camargo³, Patrícia Morais Lopes Pereira¹, Jussevania Pereira Santos², Guilherme Bartolomeu-Gonçalves^{2

4}, Lais Alonso⁵, Cesar Armando Contreras Lancheros⁶, Antonio Alonso⁵, Celso Vataru Nakamura^{1

6}, Fernando Macedo Jr³, Phileno Pinge-Filho^{1

7}, Lucy Megumi Yamauchi^{1

2}, Marcelle de Lima Ferreira Bispo³, Eliandro Reis Tavares^{1

2}, Sueli Fumie Yamada-Ogatta^{1

2

4}

Affiliations

¹ Programa de Pós-graduação em Microbiologia, Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
² Laboratório de Biologia Molecular de Microrganismos, Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
³ Laboratório de Síntese de Moléculas Medicinais, Departamento de Química, Centro de Ciências Exatas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
⁴ Programa de Pós-graduação em Fisiopatologia Clínica e Laboratorial, Departamento de Patología, Análises Clínicas e Toxicológicas, Centro de Ciências da Saúde, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
⁵ Instituto de Física, Universidade Federal de Goiás, Goiânia, Goiás, Brazil.
⁶ Laboratório de Inovação Tecnológica no Desenvolvimento de Fármacos e Cosméticos, Departamento de Ciências Básicas da Saúde, Centro de Ciências da Saúde, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.
⁷ Laboratório de Imunopatologia Experimental, Departamento de Ciências Patológicas, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.

Abstract

Introduction: Cryptococcus neoformans is one of the leading causes of invasive fungal infections worldwide. Cryptococcal meningoencephalitis is the main challenge of antifungal therapy due to high morbidity and mortality rates, especially in low- and middle-income countries. This can be partly attributed to the lack of specific diagnosis difficulty accessing treatment, antifungal resistance and antifungal toxicity.

Methods: In the present study, the effect of the synthetic thiourea derivative N-(butylcarbamothioyl) benzamide (BTU-01), alone and combined with amphotericin B (AmB), was evaluated in planktonic and sessile (biofilm) cells of C. neoformans.

Results: BTU-01 alone exhibited a fungistatic activity with minimal inhibitory concentrations (MICs) ranging from 31.25 to 62.5 μg/mL for planktonic cells; and sessile MICs ranging from 125.0 to 1000.0 μg/mL. BTU-01 caused a concentration-dependent inhibitory activity on cryptococcal urease and did not interfere with plasma membrane fluidity. Molecular docking was performed on Canavalia ensiformis urease, and BTU-01 showed relevant interactions with the enzyme. The combination of BTU-01 and AmB exhibited synergistic fungicidal activity against planktonic and sessile cells of C. neoformans. Microscopic analysis of C. neoformans treated with BTU-01, alone or combined with AmB, revealed a reduction in cell and capsule sizes, changes in the morphology of planktonic cells; a significant decrease in the number of cells within the biofilm; and absence of exopolymeric matrix surrounding the sessile cells. Neither hemolytic activity nor cytotoxicity to mammalian cells was detected for BTU-01, alone or combined with AmB, at concentrations that exhibited antifungal activity. BTU-01 also displayed drug-likeness properties.

Conclusion: These results indicate the potential of BTU-01, for the development of new strategies for controlling C. neoformans infections.

Keywords: antibiofilm; antivirulence; cryptococcosis; synergism; thiourea; urease.

Grants and funding

This study was supported by grants from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Financial Code 01). GA was funded by a graduate scholarship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). LS, MF, and GB-G were funded by a graduate scholarship from CAPES. PC and PP were funded by a post-graduate scholarship from CAPES. AA, CN, PP-F, and SY-O were funded by a research fellowship from CNPq.