Skip to main content
NCBI home page
Brazilian Journal of Microbiology logoLink to Brazilian Journal of Microbiology
. 2023 Nov 23;55(1):133–142. doi: 10.1007/s42770-023-01158-0

Synthesis and antifungal evaluation against Candida spp. of the (E)-3-(furan-2-yl)acrylic acid

Paulo César Trindade da Costa 1,✉, Thales Luciano Bezerra Santos 2, Jaqueline Ferreira Ramos 3, Jonh Anderson Macêdo Santos 3, Francinalva Dantas de Medeiros 2, Juliano Carlo Rufino Freitas 2,3, Wylly Araújo de Oliveira 2
PMCID: PMC10920609  PMID: 37995041

Abstract

Infections of fungal origin are mainly caused by Candida spp. Some species, such as C. albicans, C. glabrata, C. parapsilosis, and C. tropicalis, stand out as promoters of diseases in humans. This study evaluated the synthesis and antifungal effects of (E)-3-(furan-2-yl)acrylic acid. The synthesis of the compound showed a yield of 88%, considered high. The minimum inhibitory concentration of the synthetic compound, amphotericin B, and fluconazole isolated against four Candida species ranged from 64 to 512 μg/mL, 1 to 2 μg/mL, and 32 to 256 μg/mL, respectively. The synergistic effect of the test compound was observed when associated with amphotericin B against C. albicans and C. tropicalis, with no antagonism between the substances against any of the strains tested. The potential drug promoted morphological changes in C. albicans, decreasing the amount of resistance and virulence, and reproduction structures, such as the formation of pseudohyphae, blastoconidia, and chlamydospores. Furthermore, it was also possible to identify the fungistatic profile of the test substance by studying the growth kinetics of C. albicans. Finally, it was observed that the test compound stimulated ergosterol biosynthesis by the yeast, probably by activating microbial resistance responses.

Keywords: Antimicrobial, Candida, Morphological transition, Checkerboard, Ergosterol

Introduction

Candida spp. are the main human fungal pathogens, which cause infections in both mucous membranes and other tissues. The gender distribution is universal and varies according to geographic regions [1]. Regarding infection by species, approximately 95% of invasive Candida infections are caused by five species: C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei [2]. These infections increase morbidity and mortality, especially in immunocompromised individuals [3, 4].

Currently, the global emergence of antibiotic resistance, in addition to the reduced development of new antibiotic molecules, represents a significant public health problem, which may lead to intractable infections in the future [5]. In this way, the search for new drugs and their respective mechanisms of action becomes increasingly essential [6].

It is relevant to understand the mechanisms by which the substances act, as well as the strategies that can be adopted in the search for improvements in the fight against fungal infections. Therefore, the analysis of the ergosterol content in C. albicans represents an essential way of elucidating the mode of action of the substances, since several antifungals have the biosynthesis of ergosterol as a target. In addition, combined therapy has a high potential for fighting infections [7, 8].

Many of the compounds that have their antifungal action investigated are of semi-synthetic origin. For example, compounds derived from 2-furfuraldehyde are notably relevant, since 2-furfuraldehyde is a material that occurs frequently in nature, characterizing a low-cost raw material [9]. Additionally, the literature deals with various applications for (E)-3-(furan-2-yl)acrylic acid and its derivatives, highlighting antimicrobial [10], antiviral [11], neuropathic [12], and antitumor activities [13].

Given the high prevalence of mortality due to fungal infections [14], associated with the factors discussed above, it is necessary to investigate the antifungal activity of (E)-3-(furan-2-yl)acrylic acid, derived from 2-furfuraldehyde, on Candida spp.

Material and methods

Synthesis of the test compound

In a 100-mL round bottom flask, 2-furfuraldehyde (0.13 mL; 1.5 mmol) and piperidine (1.7 mL) were added. Later, potassium carbonate (0.134 g and 0.75 mmol), malonic acid (0.34 g and 3.3 mmol), and water (1.5 mL) were added. This mixture was heated to 90 °C and stirred for 1 h. Completion of the reaction was determined by thin-layer chromatography after 1 h of reaction. Then, the reaction mixture was neutralized with an HCl solution (1.0 M) in an ice bath resulting in the formation of a brown precipitate. The solid was filtered, washed with cold water, and recrystallized using the eluent system water–ethanol (in a 50:50 ratio), leading to (E)-3-(furan-2-yl)acrylic acid in 88% yield.

Minimum inhibitory concentration

The strains tested were C. albicans ATCC 76485, C. parapsilosis ATCC 22019, C. glabrata ATCC 90030, and C. tropicalis ATCC 13803. After cultivation, the microorganisms were kept in Sabouraud dextrose agar (SDA) under refrigeration (4 °C) during the period of carrying out the experiments, this process will occur monthly to maintain the viability of the cells.

The inoculum of each of the strains tested was prepared by adding a few colonies of the microorganism in saline solution (NaCl 0.9%) until forming a suspension with approximately 1–5 × 106 CFU/mL, adjusted according to the 0.5 of the McFarland scale. The culture medium used in the tests to evaluate the antifungal activity was Sabouraud dextrose broth, prepared according to the manufacturer’s instructions. Also, the solutions with (E)-3-(furan-2-yl)acrylic acid and antifungal agents were prepared at the time of carrying out the tests, initially dissolving them in dimethylsulfoxide (DMSO) and then having their final volume completed with sterile distilled water. Controls were performed with DMSO.

The minimal inhibitory concentrations (MIC) of (E)-3-(furan-2-yl)acrylic acid, amphotericin B, and fluconazole against Candida spp. were determined according to the microdilution method using 96-well microtiter plates [15]. In a 96-well plate, the furan compound and antifungal agents were tested at concentrations from 512 to 1 μg/mL, in serial dilutions 1:2. Plates with Candida spp., culture medium, and antifungal agents were incubated at 35 °C for 24–48 h. After which, fungal growth was observed. In addition, MIC was the lowest concentration of substances capable of inhibiting the growth of the microorganism.

MIC was considered the lowest concentration of the substance capable of inhibiting the growth of microorganisms, considering the agent and the microorganism [16]. Tests were performed in triplicate.

Morphological transition

An analysis of possible interference of the furan derivative in the morphological transition of C. albicans was carried out, using the microculture technique on a slide in a humid chamber (methodology proposed by Dalmau) [17, 18]. The cornmeal agar culture medium was added with 1% tween 80 fractionated in sterile tubes containing the tested compounds in concentrations corresponding to the MIC. After homogenization, each culture medium was placed on a glass slide. Soon after the medium solidified, the microorganism was seeded on the agar, forming two parallel streaks, then a coverslip was inserted to cover the microculture, and the plates were incubated at 35 ± 2 °C for 48–72 h, after which the plates Petri dishes were incubated at 25 ± 2 °C for another 48–72 h. In this way, we obtained the negative control and positive controls (AFA, amphotericin B, and fluconazole) in concentrations corresponding to the MIC and MIC/2. The slides were analyzed daily for five days by optical microscopy using a microscope, at a magnification of 400x, to observe the absence or presence of characteristic structures.

Time-kill assay

In the study of the interference of the compounds on the growth curve of C. albicans ATCC 76485, the minimum and multiples inhibitory concentrations of (E)-3-(furan-2-yl)acrylic acid, amphotericin B, and fluconazole were tested [19].

Initially, in a ratio of 1:10, a yeast suspension in Saboraud dextrose broth (SDB) with the test compound, amphotericin B, and fluconazole was inoculated, in addition to sterility and fungal viability controls. After incubation, at intervals of 0 h, 4 h, 8 h, and 24 h, an aliquot of 10 μL of the solution was uniformly seeded on SDA plates. Inoculated plates were incubated at 35 ± 2 °C for 24–48 h. Then, the count of CFU present in the Petri dishes was performed, and the amount per CFU/mL of solution was determined in each period for each substance and its concentrations. The experiment was carried out in triplicate.

Analysis of ergosterol

To analyze the activity of (E)-3-(furan-2-yl)acrylic acid on ergosterol biosynthesis in C. albicans, the microorganism was inoculated in tubes containing SDB, with the addition of the respective molecules tested at different concentrations (MICx2 and MIC). Cultures were incubated at 35 °C for 24 h. After this period, the tubes were centrifuged at 3000 rpm for 15 min, and then, the wet weight of the sediments was determined. Subsequently, 3 mL of a 25% alcoholic potassium hydroxide solution (25 g of KOH and 36 mL of sterile distilled water brought to 100 mL with 100% ethanol). The suspensions were then incubated in a water bath at 80 °C for 1 h. After that, a mixture of 1 mL of water and 3 mL of heptane was added, followed by vigorous vortex mixing for 5 min. Finally, the heptane layer was filtered, with a 0.45-µm syringe filter, and transferred to properly identified vials, for subsequent quantification of ergosterol by high-performance liquid chromatography (HPLC) [20].

The quantification of ergosterol was performed in HPLC equipment (Shimadzu) with a UV detector with a diode arrangement, scanning in the UV–visible region, and monitoring the wavelength of 282 nm. Analyses were performed in isocratic mode, with methanol as a mobile phase at a flow rate of 1.4 mL/min. As a stationary phase, a C18 Shim-pack CLC-ODS analytical column (250 × 4.6 mm ID) was used, with a particle size of 5 μm, under a temperature of 30 °C.

The chromatographic analysis was developed and validated using the standard addition method. The parameters used were specificity, linearity, precision, and accuracy. Ergosterol (Sigma Chemical) was used as the analytical standard, and the analyses were carried out in triplicate on an inter- and intra-day [21].

Checkerboard assay

After obtaining MIC values, the association of (E)-3-(furan-2-yl)acrylic acid with amphotericin B and fluconazole against microorganisms was determined through the fractional inhibitory concentration index (FICI) using the checkerboard microdilution method according to MIC [22]. The concentration of each compound in the combination varied between MIC/8 and MICx8 and was diluted in a 1:2 ratio. Initially, 100 μL of the culture medium was added to the wells of the microdilution plates. Subsequently, 50 μL of each substance tested at different concentrations was added vertically (amphotericin B or fluconazole) and horizontally (E)-3-(furan-2-yl)acrylic acid into the wells of plates [23]. The FICI is calculated by the following formula: FICI = (MICA in combination/MICA alone) + (MICB in combination/MICB alone) [22].

Statistical analysis

Data obtained from MIC, association, and micromorphology assays were analyzed using descriptive statistics. The growth kinetics curve was plotted by Log10 CFU/mL as a function of time (hours) and concentrations of the tested compounds and for the statistical analysis the Kruskal–Wallis test was used. In the evaluation of the ergosterol content, the Kruskal–Wallis test was used. the ANOVA. Data were considered significant when p < 0.05.

Results

(E)-3-(Furan-2-yl)acrylic acid was synthesized from the reaction between 2-furfuraldehyde and malonic acid (Fig. 1). This compound was obtained as a brown crystalline solid in 88% yield. Additionally, the chemical shift data for the hydrogen and carbon-13 nuclei of (E)-3-(furan-2-yl)acrylic acid are described in Table 1.

Fig. 1.

Fig. 1

Open in a new tab

1H NMR and 13C NMR spectroscopic data of (E)-3-(furan-2-yl)acrylic acid

Table 1.

1H NMR and 13C NMR spectroscopic data of (E)-3-(furan-2-yl)acrylic acid

graphic file with name 42770_2023_1158_Tab1_HTML.jpg

Open in a new tab

bsbroad singlete,ddoublet, dddoublet and doublet,Jcoupling constant

The antifungal activities of (E)-3-(furan-2-yl)acrylic acid, amphotericin B, and fluconazole were investigated (Table 2). Analyzing Table 2, it was possible to verify that there was divergence in the growth of Candida strains according to the antifungal compounds and their different concentrations.

Table 2.

MIC of antifungals for Candida spp

Antifungals C. albicans ATCC 76485 C. glabrata ATCC 90030 C. parapsilosis ATCC 22019 C. tropicalis ATCC 13803
(E)-3-(Furan-2-yl)acrylic acid 64 μg/mL 256 μg/mL 512 μg/mL 256 μg/mL
Amphotericin B 1 μg/mL 2 μg/mL 1 μg/mL 1 μg/mL
Fluconazole 256 μg/mL 256 μg/mL 64 μg/mL 32 μg/mL
Open in a new tab

(E)-3-(Furan-2-yl)acrylic acid MICs against Candida spp ranged from 64 to 512 μg/mL, showing greater efficacy against C. albicans when compared to other strains tested. The MICs of amphotericin B were lower than those of the synthetic compound against all tested strains, varying between 1 and 2 μg/mL. Fluconazole, compared to (E)-3-(furan-2-yl)acrylic acid, had the highest MIC values against C. albicans, the same against C. parapsilosis, and lowest compared to the other strains tested, ranging from 32 to 256 μg/mL (Fig. 2).

Fig. 2.

Fig. 2

Open in a new tab

Magnification of the 1H NMR spectrum of (E)-3-(furan-2-yl)acrylic acid to determine double bond configuration

(E)-3-(Furan-2-yl)acrylic acid was effective in reducing the development of virulence structures such as pseudohyphae, blastoconidia, and chlamydoconidia when compared to the control. When the microorganism was cultured on cornmeal agar in the absence of any drug, pseudohyphae with blastoconidia, with thick-walled terminal chlamydoconidia, was observed. Amphotericin B and fluconazole were more effective than the (E)-3-(furan-2-yl)acrylic acid in inhibiting the formation of virulence structures (Fig. 3).

Fig. 3.

Fig. 3

Open in a new tab

Effect of (E)-3-(furan-2-yl)acrylic acid and controls on the micromorphology of C. albicans ATCC 76485 after 4 days of cultivation. a Micromorphology of the microorganism without the test substance. b Micromorphology of the microorganism when exposed to amphotericin B at its MIC. c Micromorphology of the microorganism when exposed to amphotericin B at its MIC/2. d Micromorphology of the microorganism when exposed to fluconazole at its MIC. e Micromorphology of the microorganism when exposed to fluconazole at its MIC/2. f Micromorphology of the microorganism when exposed to (E)-3-(furan-2-yl)acrylic acid in its MIC. g Micromorphology of the microorganism when exposed to (E)-3-(furan-2-yl)acrylic acid at its MIC/2

Figure 4 contains the results of the number of log CFU/mL according to the time intervals for the concentrations used on C. albicans. Thus, it was possible to observe a similar behavior between the test compound and fluconazole over time, while amphotericin B showed a different pattern.

Fig. 4.

Fig. 4

Open in a new tab

Viability of C. albicans ATCC 76485 when exposed to (E)-3-(furan-2-yl)acrylic acid and controls. Flu: fluconazole; Anf: amphotericin B; AFA: (E)-3-(furan-2-yl)acrylic acid

For the chromatographic method of quantification of ergosterol, a linear calibration model was established, using the standard addition method. The method of proved to be linear in the range of 0.63 to 10 μg/mL, with a coefficient of determination R2 equal to 0.9738. The results found for repeatability and precision show that the ergosterol quantification method provides concordant results. The coefficients of variation (CV%) obtained were mostly below 20%, given the complexity of the biological matrix [24].

The ergosterol retention time was approximately 8.79 min (Fig. 5). The quantification of ergosterol showed that the content of this sterol in yeast decreased significantly after the control treatment with fluconazole in MICx2 and MIC, decreasing respectively by 69% and 79%. With (E)-3-(furan-2-yl)acrylic acid, there was an increase, although not statistically significant, in the ergosterol content in the MIC and MICx2, increasing its production by 89% and 15% in that order, concerning the control (p < 0.05) (Fig. 6).

Fig. 5.

Fig. 5

Open in a new tab

Examples of chromatograms of the ergosterol pattern and the microorganism when exposed to the tested substances. Chromatogram 1: ergosterol standard; Chromatogram 2: microorganism without exposure to the substance; Chromatogram 3: microorganism exposed to fluconazole (MICx2); Chromatogram 4: microorganism exposed to (E)-3-(furan-2-yl)acrylic acid (MICx2

Fig. 6.

Fig. 6

Open in a new tab

Quantitative analysis of ergosterol content in Candida albicans after treatment with different concentrations of fluconazole or (E)-3-(furan-2-yl)acrylic acid. MO: microorganism; Flu: fluconazole; AFA: (E)-3-(furan-2-yl)acrylic acid. *p < 0.05 compared to the control (ANOVA–Dunnett’s)

When (E)-3-(furan-2-yl)acrylic acid was tested in association with amphotericin B or with fluconazole against Candida spp., the results ranged from synergistic to indifferent, and there was no antagonistic interaction (Table 3).

Table 3.

Associations between (E)-3-(furan-2-yl)acrylic acid and commercial antifungals for Candida spp.—type of interaction

Microorganism Antifungals FIC A FIC B FICI Type of interaction
C. albicans AFA + ANFB 0.125 0.25 0.375 Synergism
ATCC 76485 AFA + FLU 1.0 0.25 1.25 Indifference
C. glabrata AFA + ANFB 2.0 0.5 2.5 Indifference
ATCC 90030 AFA + FLU 2.0 1.0 3.0 Indifference
C. parapsilosis AFA + ANFB 0.25 0.5 0.75 Addition
ATCC 22019 AFA + FLU 0.5 0.25 0.75 Addition
C. tropicalis APFA + ANFB 0.125 0.25 0.375 Synergism
ATCC 13803 AFA + FLU 0.5 0.5 1.0 Indifference
Open in a new tab

AFA(E)−3−(furan−2−yl)acrylic acid,ANFBamphotericin B,FLUfluconazole,FICfractional inhibitory concentration,FICIfractional inhibitory concentration index

Discussion

In this study, the compound was obtained in high yield and we observed that (E)-3-(furan-2-yl)acrylic acid has an antifungal effect on Candida spp., with the ability to inhibit the morphological transition in C. albicans. In addition, the compound test has a fungistatic character, acting in a similar way to fluconazole, and, perhaps because of this, it can increase ergosterol biosynthesis in C. albicans in response to the non-lethal stress of the compound when administered in the MIC. Still, when (E)-3-(furan-2-yl)acrylic acid is used in association with amphotericin B, the effects are synergistic against C. albicans and C. tropicalis.

(E)-3-(Furan-2-yl)acrylic acid was synthesized through condensation idealized by Emil Knoevenagel and later modified by Oscar Doebner, in which a carboxylic acid reacts with an aldehyde, in a basic medium, leading to the formation of an a,b-unsaturated compound (Fig. 1) [25].

The Knoevenagel-Doebner condensation, in general, is an efficient and selective method, leading to the desired products in high yields [26, 27], specifically, the synthesized (E)-3-(furan-2-yl)acrylic acid was obtained with 88% of yield, a value close to that obtained by Kalyaev et al. 2022 [10].

The chemical shift data for hydrogen and carbon-13 nuclei obtained are similar to those described by Kalyaev et al. (2022) [10]. Additionally, by expanding the 1H NMR spectrum of (E)-3-(furan-2-yl)acrylic acid, the signals referring to olefinic protons 2 and 3 and their corresponding coupling constants are observed, indicating the exclusive formation of the E isomer (Fig. 6).

Although the existence of different MIC evaluation methods and variations in experimental conditions contribute to making the comparison between some substances difficult, determining the lowest concentration of the compound capable of completely inhibiting microbial growth is an interesting starting point for research [28].

Several substances exert antimicrobial effects, whether they are synthetic or natural products. The furan derivative synthesized in the laboratory called methyl-5-(hydroxymethyl)-2-furan carboxylate had its antimicrobial effect demonstrated by Phutdhawong et al. [29] also by the broth microdilution method, however, against bacteria. The antifungal potential of the alcoholic extract of Curcuma longa against Candida spp. was evidenced by Murugesh et al. [30].

In this study, (E)-3-(furan-2-yl)acrylic acid was more effective against C. albicans, and similar potential, in terms of MIC, to fluconazole was observed. D’arrigo et al. [31] observed the fungicidal activity of pistachio bark essential oil at concentrations between 2.5 and 5.0 mg/mL.

The morphological transition capacity of C. albicans is characterized as an essential virulence factor since different functions such as dissemination and adhesion occurring in yeast cells and pseudohyphae increase pathogenicity. In this sense, the study of morphological changes caused by (E)-3-(furan-2-yl)acrylic acid is of great relevance in detailing its antifungal activity [32]. In the present research, (E)-3-(furan-2-yl)acrylic acid reduced the development of virulence structures such as pseudohyphae, blastoconidia, and claminoconidia compared to the control with the microorganism cultured in the absence of antifungal agents.

Many other substances act by inhibiting the morphological transition in C. albicans. Kalopanaxsaponin A effectively inhibited the hyphal transition for 4 h at a concentration of 12 μg/mL, and this effect was related to farnesol secretion, an important quorum-sensing molecule involved in preventing morphological transformation [33]. The study by Xu et al. [34] demonstrated that, with the use of 2.0 mg/mL, coumarin was effective in inhibiting hyphal formation in C. albicans, almost completely blocking this transition, regardless of the dose. β-Citronellol is also part of the range of substances capable of inhibiting this transition [35].

Thus, (E)-3-(furan-2-yl)acrylic acid appears as a potential morphological transition inhibitor. However, clinical studies need to be developed to obtain more details related to its promising ability to attenuate infections caused by C. albicans.

(E)-3-(Furan-2-yl)acrylic acid contributed to the decrease in C. albicans compared to the control without the addition of antifungals. However, a 99.9% reduction in the number of microorganisms was not observed, which characterizes the fungistatic profile of the test substance, acting similarly to fluconazole.

An antifungal compound can exert fungistatic or fungicidal activity, and factors such as the concentration and time of exposure of the drug to the microorganism are determinants. Also, defining whether the substance is fungistatic or fungicidal is fundamental for the analysis of therapeutic feasibility, since the responses of immunocompromised and immunocompetent individuals to the treatment of infections depend strictly on the profile of the drug administered [36].

Many and varied substances act by partially inhibiting fungal growth, such as Loureirin A, which was able to inhibit fungal growth after 24 h, but without statistical significance [37]. Miao et al. [38] observed a fungistatic effect of basil oil (Ocimum basilicum) on C. albicans. The effect of the anidulafugin on the growth of 14 strains of C. albicans was tested, showing a fungicidal effect against 2 strains and a fungistatic effect against the others [39]. Still, the fungicidal effect is essential and quite common in the literature, an example would be the ability of magnesium oxide nanoparticles to significantly increase the number of yeast cells after 24 h [40]. In this way, this test allows obtaining details that can be used in an eventual research that aims to evaluate the clinical use of this antifungal.

The ability of (E)-3-(furan-2-yl)acrylic acid to increase the level of ergosterol in C. albicans cells was identified in both MIC (+ 89%) and MICx2 (+ 15%), but sterol production occurred at a lower rate when drug concentration was increased. This fact can be attributed to a resistance mechanism developed by the yeast in response to the stress caused by the drug, which may be related to its fungistatic character. There is probably a greater expression of the genes that encode the biosynthesis of ergosterol since its concentration plays an essential role in the integrity of the cell membrane [41, 42].

Ergosterol biosynthesis, which occurs in the endoplasmic reticulum through the coordinated action of approximately 25 enzymes, is one of the main cellular pathways targeted by antifungal compounds [43]. Some studies indicate that the transcription factor for apoptosis (Upc2) in C. albicans detects the level of intracellular ergosterol and generates a response in the activation of genes necessary for the biosynthesis of this sterol. Thus, ergosterol depletion caused by azoles or other classes of antifungals can activate this Upc2-mediated transcriptional response [44].

Thus, several mechanisms induced by antifungals may contribute to the activation of genes that encode ergosterol biosynthesis. However, experimental data are still needed to reach a definitive conclusion [45].

Varied strategies are used to optimize the treatment of the increasing incidence of infections caused by Candida spp., among them is the combined therapy, which aims to reduce the risk of antifungal resistance to monotherapy, decrease possible side effects, and reduce treatment time. Therefore, the search for synergistic interactions between substances is an essential tool in the fight against fungal infections [46].

In this study, synergistic interactions of (E)-3-(furan-2-yl)acrylic acid in association with amphotericin B against C. albicans and C. tropicalis were observed. This discovery was important because although amphotericin B is the antifungal with the greatest fungicidal activity, its use has some limitations due to its toxicity [47]. Other studies have also found substances capable of interacting synergistically with amphotericin B. Erythromycin, for example, increases the activity of amphotericin B against varied species of Candida [47]. The combination of chitooligosaccharides with amphotericin B against Candida strains also represents new therapeutic perspectives for the treatment of candidiasis [48]. In this way, these data encourage the constant search for new antifungal agents and combinations among them that aim at therapeutic success, considering specific treatment factors, a factor that in light of the divergent responses discussed above, should not be neglected.

Given the facts discussed above that characterize (E)-3-(furan-2-yl)acrylic acid as a promising antifungal agent, we suggest conducting in vivo tests to evolve its knowledge in the pharmacological field. However, we believe this study helps understand the antifungal activity of this compound against Candida spp.

Conclusion

In this study, (E)-3-(furan-2-yl)acrylic acid was synthesized from the reaction between 2-furfuraldehyde and malonic acid, and their synthesis had a high yield. Our study expresses great utility in understanding the antifungal activity of the (E)-3-(furan-2-yl)acrylic acid against Candida spp. In addition, the substance may also serve as a starting point for the synthesis of more potent molecules. Finally, we believe that carrying out new experiments in further studies is essential to broaden the understanding of the effects of this compound.

Author contribution

Conceptualization: PCTdC, JCRdF, and WAdO. Formal analysis and investigation: PCTdC and WAdO. Methodology: PCTdC, TLBS, JFR, JAMS, and FDdM. Writing—original draft preparation: PCTdC. Writing—review and editing: JCRdF, FDdM, and WAdO. Supervision: JCRdF and WAdO. All authors gave final approval and agreed to be accountable for all aspects of the work ensuring integrity and accuracy.

Funding

This study was funded by Paraiba State Research Foundation (FAPESQ, 3157/2021) and National Council for Scientific and Technological Development (CNPq, 434012/2018–1). The authors thank Coordination for the Improvement of Higher Education Personnel (CAPES) for the scholarships granted.

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Declarations

Conflicts of interest

The authors declare no competing interests.

Footnotes

Responsible Editor: Celia Maria de Almeida Soares

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  • 1.Gonzalez-Lara MF, Ostrosky-Zeichner L. Invasive candidiasis. Semin Respir Crit Care Med. 2020;41:03–12. doi: 10.1055/s-0040-1701215. [DOI] [PubMed] [Google Scholar]
  • 2.Quindós G, Marcos-Arias C, San-Millán R, Mateo E, Eraso E. The continuous changes in the aetiology and epidemiology of invasive candidiasis: from familiar Candida albicans to multiresistant Candida auris. Int Microbiol. 2018;21:107–119. doi: 10.1007/s10123-018-0014-1. [DOI] [PubMed] [Google Scholar]
  • 3.Thompson GR, et al. Fungal endocarditis: pathophysiology, epidemiology, clinical presentation, diagnosis, and management. Clin Microbiol Rev. 2023;13:e0001923. doi: 10.1128/cmr.00019-23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Robbins N, Wright GD, Cowen LE. Antifungal drugs: the current armamentarium and development of new agents. Microbiol Spectr. 2016;4:903–922. doi: 10.1128/microbiolspec.FUNK-0002-2016. [DOI] [PubMed] [Google Scholar]
  • 5.Varaldo PE, et al. The first outstanding 50 years of “Università Politecnica delle Marche”. Cham. Switzerland: Springer; 2020. Antimicrobial resistance: a challenge for the future; pp. 13–29. [Google Scholar]
  • 6.Revie NM, Iyer KR, Robbins N, Cowen LE. Antifungal drug resistance: evolution, mechanisms and impact. Curr Opin Microbiol. 2018;45:70–76. doi: 10.1016/j.mib.2018.02.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Kibbler CC. The Pro-debate: how can we improve the outcome of invasive fungal infection? The case for combination therapy. Infection. 2012;16:3–10. doi: 10.1016/S0123-9392(12)70021-8. [DOI] [Google Scholar]
  • 8.Tavakkoli A, Johnston TP, Sahebkar A. Antifungal effects of statins. Pharmacol Ther. 2020;208:107483. doi: 10.1016/j.pharmthera.2020.107483. [DOI] [PubMed] [Google Scholar]
  • 9.Binder JB, Blank JJ, Cefali AV, Raines RT. Synthesis of furfural from xylose and xylan. Chemsuschem. 2010;3:1268–1272. doi: 10.1002/cssc.201000181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Kalyaev MV, et al. Synthesis of 3-Aryl-3-(furan-2-yl)propanoic acid derivatives, and study of their antimicrobial activity. Molecules. 2022;19:4612. doi: 10.3390/molecules27144612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Emad ME, Ghanem MS, El-Essawy AF, El-Sayed FA. Synthesis and in vitro antiviral evaluation of novel 1-arylpyrazoles and their N-and S-Glycosides. Lett Drug Des Discov. 2011;8:882–889. doi: 10.2174/1570180811108090822. [DOI] [Google Scholar]
  • 12.Arias HR, et al. (E)-3-Furan-2-yl-N-p-tolyl-acrylamide and its derivative DM489 decrease neuropathic pain in mice predominantly by α7 nicotinic acetylcholine receptor potentiation. ACS Chem Neurosci. 2020;11:3603–3614. doi: 10.1021/acschemneuro.0c00476. [DOI] [PubMed] [Google Scholar]
  • 13.Swamy PV, et al. Synthesis, biological evaluation and molecular docking studies of some novel cyclopropane carbohydrazide derivatives as potential anticancer agents. J Chem Sci. 2016;128:929–939. doi: 10.1007/s12039-016-1081-5. [DOI] [Google Scholar]
  • 14.Keighley C, et al. Candidaemia and a risk predictive model for overall mortality: a prospective multicentre study. BMC Infect Dis. 2019;19:445. doi: 10.1186/s12879-019-4065-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: a review. J Pharm Anal. 2016;6:71–79. doi: 10.1016/j.jpha.2015.11.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Wiederhold NP. Antifungal susceptibility testing: a primer for clinicians. Open Forum Infect Dis. 2021;8:1–13. doi: 10.1093/ofid/ofab444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Sidrim JJC, Rocha MFG. Micologia médica à luz de autores contemporâneos. São Paulo: Guanabara Koogan; 2004. p. 396. [Google Scholar]
  • 18.Walsh C. Opinion—anti-infectives: where will new antibiotics come from? Nat Rev Microbiol. 2003;1:65–70. doi: 10.1038/nrmicro727. [DOI] [PubMed] [Google Scholar]
  • 19.Klepser ME, Ernst EJ, Lewis RE, Ernst ME, Pfaller MA. Influence of test conditions on antifungal time-kill curve results: proposal for standardized methods. Antimicrob Agents Chemother. 1998;42:1207–1212. doi: 10.1128/AAC.42.5.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Arthington-Skaggs BA, Warnock DW, Morrison CJ. Quantitation of Candida albicans ergosterol content improves the correlation between in vitro antifungal susceptibility test results and in vivo outcome after fluconazole treatment in a murine model of invasive candidiasis. Antimicrob Agents Chemother. 2000;44:2081–2085. doi: 10.1128/AAC.44.8.2081-2085.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Cabral ME, Figueroa LIC, Fariña JI. Synergistic antifungal activity of statin–azole associations as witnessed by Saccharomyces cerevisiae-and Candida utilis-bioassays and ergosterol quantification. Rev Iberoam Micol. 2013;30:31–38. doi: 10.1016/j.riam.2012.09.006. [DOI] [PubMed] [Google Scholar]
  • 22.Cuenca-Estrella M. Combinations of antifungal agents in therapy–what value are they? J Antimicrob Chemother. 2004;54:854–869. doi: 10.1093/jac/dkh434. [DOI] [PubMed] [Google Scholar]
  • 23.Correa-Royero J, Tangarife V, Durán C, Stashenko E, Mesaarango A. In vitro antifungal activity and cytotoxic effect of essential oils and extracts of medicinal and aromatic plants against Candida krusei and Aspergillus fumigatus. Rev Bras Farmacogn. 2010;20:734–741. doi: 10.1590/S0102-695X2010005000021. [DOI] [Google Scholar]
  • 24.Chiocchio VM, Matković L. Determination of ergosterol in cellular fungi by HPLC. A modified technique. J Arg Chem Soc. 2011;98:10–15. [Google Scholar]
  • 25.Van Beurden K, Koning S, Molendijk D, Van Schijndel J. The Knoevenagel reaction: a review of the unfinished treasure map to forming carbon–carbon bonds. Green Chem Lett Rev. 2020;13:349–364. doi: 10.1080/17518253.2020.1851398. [DOI] [Google Scholar]
  • 26.Zacuto MJ. Synthesis of acrylamides via the Doebner-Knoevenagel condensation. J Org Chem. 2019;84:6465–6474. doi: 10.1021/acs.joc.9b00450. [DOI] [PubMed] [Google Scholar]
  • 27.Lu J, Toy PH. Organocatalytic decarboxylative Doebner-Knoevenagel reactions between arylaldehydes and monoethyl malonate mediated by a bifunctional polymeric catalyst. Synlett. 2011;12:1723–1726. doi: 10.1055/s-0030-1260808. [DOI] [Google Scholar]
  • 28.Van De Vel E, Sampers I, Raes K. A review on influencing factors on the minimum inhibitory concentration of essential oils. Crit Rev Food Sci Nutr. 2019;59:357–378. doi: 10.1080/10408398.2017.1371112. [DOI] [PubMed] [Google Scholar]
  • 29.Phutdhawong W, Inpang S, Taechowisan T, Phutdhawong WS. Synthesis and biological activity studies of methyl-5-(hydroxymethyl)-2-furan carboxylate and derivatives. Orient J Chem. 2019;35:1080–1085. doi: 10.13005/ojc/350322. [DOI] [Google Scholar]
  • 30.Murugesh J, Annigeri RG, Mangala GK, Mythily PH, Chandrakala J. Evaluation of the antifungal efficacy of different concentrations of Curcuma longa on Candida albicans: an in vitro study. J Oral Maxillofac Pathol. 2019;23:305. doi: 10.4103/jomfp.JOMFP_200_18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.D'arrigo M, Bisignano C, Irrera P, Smeriglio A, Zagami R, Trombetta D, Romeo O, Mandalari G. In vitro evaluation of the activity of an essential oil from Pistacia vera L. variety Bronte hull against Candida spp. BMC Complement Altern Med. 2019;19:1–6. doi: 10.1186/s12906-018-2425-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Jacobsen ID, Wilson D, Wächtler B, Brunke S, Naglik JR, Hube B. Candida albicans dimorphism as a therapeutic target. Expert Rev Anti Infect Ther. 2012;10:85–93. doi: 10.1586/eri.11.152. [DOI] [PubMed] [Google Scholar]
  • 33.Li Y, et al. Anticandidal activity of Kalopanaxsaponin A: effect on proliferation, cell morphology, and key virulence attributes of Candida albicans. Front Microbiol. 2019;10:2844. doi: 10.3389/fmicb.2019.02844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Xu K, Wang JL, Chu MP, Jia C. Activity of coumarin against Candida albicans biofilms. J Mycol Med. 2019;29:28–34. doi: 10.1016/j.mycmed.2018.12.003. [DOI] [PubMed] [Google Scholar]
  • 35.Sharma Y, Rastogi SK, Perwez A, Rizvi MA, Manzoor N. β-citronellol alters cell surface properties of Candida albicans to influence pathogenicity related traits. Med Mycol J. 2020;58:93–106. doi: 10.1093/mmy/myz009. [DOI] [PubMed] [Google Scholar]
  • 36.Siddiqui ZN, Farooq F, Musthafa TM, Ahmad A, Khan AU. Synthesis, characterization and antimicrobial evaluation of novel halopyrazole derivatives. J Saudi Chem Soc. 2013;17:237–243. doi: 10.1016/j.jscs.2011.03.016. [DOI] [Google Scholar]
  • 37.Mei-Yu L, et al. Effect of loureirin A against Candida albicans biofilms. Chin J Nat Med. 2019;17:616–623. doi: 10.1016/S1875-5364(19)30064-0. [DOI] [PubMed] [Google Scholar]
  • 38.Miao Q, et al. Microbial metabolomics and network analysis reveal fungistatic effect of basil (Ocimum basilicum) oil on Candida albicans. J Ethnopharmacol. 2020;260:113002. doi: 10.1016/j.jep.2020.113002. [DOI] [PubMed] [Google Scholar]
  • 39.Gil-Alonso S, Quindós G, Eraso E, Jauregizar N. Postantifungal effect of anidulafungin against Candida albicans, Candida dubliniensis, Candida africana, Candida parapsilosis, Candida metapsilosis and Candida orthopsilosis. Rev Esp Quimioter. 2019;32:183–188. [PMC free article] [PubMed] [Google Scholar]
  • 40.Kong F, et al. Antifungal activity of magnesium oxide nanoparticles: effect on the growth and key virulence factors of Candida albicans. Mycopathologia. 2020;185:485–494. doi: 10.1007/s11046-020-00446-9. [DOI] [PubMed] [Google Scholar]
  • 41.Mutasa T, Mangoyi R, Mukanganyama S. The effects of Combretum zeyheri leaf extract on ergosterol synthesis in Candida albicans. J Herbs Spices Med Plants. 2015;21:211–217. doi: 10.1080/10496475.2014.941451. [DOI] [Google Scholar]
  • 42.Lee Y, Puumala E, Robbins N, Cowen LE. Molecular mechanisms in Candida albicans and beyond. Chem Rev. 2020;121:3390–3411. doi: 10.1021/acs.chemrev.0c00199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Rodrigues ML. The multifunctional fungal ergosterol. MBio. 2018;9:01755–1818. doi: 10.1128/mBio.01755-18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Yang H, Tong J, Lee CW, Ha S, Eom SH, Im YJ. Structural mechanism of ergosterol regulation by fungal sterol transcription factor Upc2. Nat Commun. 2015;6:1–13. doi: 10.1038/ncomms7129. [DOI] [PubMed] [Google Scholar]
  • 45.Hu C, Zhou M, Wang W, Sun X, Yarden O, Li S. Abnormal ergosterol biosynthesis activates transcriptional responses to antifungal azoles. Front Microbiol. 2018;9:1–9. doi: 10.3389/fmicb.2018.00009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Livengood SJ, Drew RH, Perfect JR. Combination therapy for invasive fungal infections. Curr Fungal Infect Rep. 2020;14:1–10. doi: 10.1007/s12281-020-00369-4. [DOI] [Google Scholar]
  • 47.Rossi AS, et al. Identification of off-patent drugs that show synergism with Amphotericin B or that present antifungal action against Cryptococcus neoformans and Candida spp. Antimicrob Agents Chemother. 2020;64:01921–2019. doi: 10.1128/AAC.01921-19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.Ganan M, Lorentzen SB, Aam BB, Eijsink VG, Gaustad P, Sørlie M. Antibiotic saving effect of combination therapy through synergistic interactions between well-characterized chito-oligosaccharides and commercial antifungals against medically relevant yeasts. PLoS ONE. 2019;14:1–13. doi: 10.1371/journal.pone.0227098. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Articles from Brazilian Journal of Microbiology are provided here courtesy of Brazilian Society of Microbiology

RESOURCES