Sains Malaysiana 49(10)(2020): 2465-2476

http://dx.doi.org/10.17576/jsm-2020-4910-12

Time-Kill Assay of N-(2-Bromoethyl)-7-Chloroquinilin-4-Amine (ACP 4A) with Fungistatic Activity against Aspergillus fumigatus

(Asai Masa Kematian Sebatian N-(2-Bromoetil)-7-Kloroquinilin-4-Amina (ACP 4A) dengan Aktiviti Fungistatiknya terhadap Aspergillus fumigatus)

DAYANG FREDALINA BASRI¹, KUEK SZE YEE¹, JACINTA SANTHANAM², MOHD ASYRAF SHAMSUDIN³, NUR HANIS ZAKARIA³, JALIFAH LATIP³ & NURUL IZZATY HASSAN³*

¹Centre for Diagnostic, Therapeutic & Investigative Studies (CODTIS), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

²Centre for Toxicology and Health Risk Studies (CORE), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

³Department of Chemical Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Received: 27 August 2019/Accepted: 4 May 2020

ABSTRACT

Six pyrano[2,3-c]pyrazole-3-carboxylate and quinoline derivatives (ACP 1A, ACP 1B, ACP 1C, ACP 1D, ACP 2A and ACP 4A) synthesized and screened for antimicrobial activity against two selected Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), three Gram-negative bacteria (Klebsiella pneumonia, Pseudomonas aeruginosa, and Escherichia coli), two yeast strains (Candida albicans and Candida glabrata) and two filamentous fungi (Aspergillus niger and Aspergillus fumigatus) using agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) of the compounds were determined using broth microdilution and streak plate method, respectively. Time-kill assay (TKA) analysis of N-(2-bromoethyl)-7-chloroquinilin-4-amine (ACP 4A) was subsequently conducted throughout the 24 h incubation period against A. fumigatus. Only ACP 4A was chosen for the evaluation of TKA analysis because the broth dilution assay confirmed that it is the most potent compound with antifungal activity. E. coli was the only one found to be susceptible to the ACP 4A compound at 20 and 30 mg mL^-1 with an inhibition zone value of 15.00 ± 0.00 and 19.00 ± 0.00 mm, respectively. MIC values of ACP 4A compound against S. aureus and C. albicans were 800 µg mL^-1. A. niger and A. fumigatus exhibited the highest susceptibility towards ACP 4A with the same MIC value (200 µg mL^-1).  ACP 4A showed fungicidal effect against C. glabrata with the same MIC and MFC values at 800 µg mL^-1. From TKA analysis, ACP 4A at 1 × MIC and 2 × MIC did not show fungicidal activity against A. fumigatus. In conclusion, ACP 4A has the potential to be developed as a fungistatic (non-fungicidal) antifungal agent, which is an advantage compared to other known fungicidal compounds, particularly in patients with a healthy immune system.

Keywords: Antimicrobial; bacteria; fungi; pyrano[2,3-c] pyrazole-3-carboxylate; quinoline

ABSTRAK

Enam terbitan kumpulan pirano[2,3-c]pirazola-3-karboksil dan kuinolina (ACP 1A, ACP 1B, ACP 1C, ACP 1D, ACP 2A, dan ACP 4A) telah disaring untuk aktiviti antimikrob ke atas dua bakteria Gram positif (Staphylococcus aureus dan Bacillus subtilis), tiga bakteria Gram negatif (Klebsiella pneumoniae, Pseudomonas aeruginosa dan Escherichia coli), dua strain yis (Candida albicans dan Candida glabrata) dan dua spesies fungus berfilamen (Aspergillus niger dan Aspergillus fumigatus) menggunakan kaedah resapan telaga. Nilai kepekatan perencatan minimum (MIC) dan kepekatan bakterisid/fungisid minimum (MBC/MFC) sebatian ditentukan menggunakan kaedah mikropencairan kaldu dan teknik coretan plat. Asai masa kematian (TKA) sepanjang tempoh inkubasi selama 24 jam untuk sebatian N-(2-Bromoetil)-7-klorokuinilin-4-amina (ACP 4A) ditentukan ke atas A. fumigatus. Hanya E. coli sahaja didapati rentan terhadap sebatian ACP 4A pada 20 dan 30 mg mL^-1 dengan nilai zon perencatan masing-masing, 15.00 ± 0.00 dan 19.00 ± 0.00 mm. Nilai MIC sebatian ACP 4A terhadap S. aureus dan C. albicans adalah 800 µg mL^-1. A. niger dan A. fumigatus menunjukkan kerentanan yang paling tinggi terhadap sebatian ACP 4A dengan nilai MIC yang sama (200 µg mL^-1). Sebatian ACP 4A menunjukkan kesan fungisid ke atas C. glabrata dengan nilai MIC dan MFC yang sama iaitu 800 µg mL^-1. Daripada analisis TKA, ACP 4A pada 1 × MIC dan 2 × MIC tidak menunjukkan aktiviti fungisid terhadap A. fumigatus. Kesimpulannya, sebatian ACP 4A berpotensi untuk dimajukan sebagai agen antikulat fungistatik yang merupakan kelebihan berbanding dengan agen fungisid yang lain, terutamanya pada pesakit dengan immunisasi yang baik.

Kata kunci: Antimikrob; bakteria; kuinolina; kulat; pirano[2,3-c]pirazola-3-karboksil

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*Corresponding author; email: drizz@ukm.edu.my