← Back

Coumarin-modified ruthenium complexes: Synthesis, characterization, and antiproliferative activity against human cancer cells.

PMID: 38864840
Archiv der Pharmazie FULL PAPER Structure Characterization and Antibacterial, Antifungal, and Antiquorum‐Sensing Activity of New Metabolites From the Lipophilic Fractions of Platycodon grandiflorum Root Hyukbean Kwon1 | Jimin Moon1 | Jeong‐Hyeon Kim2 | Prima F. Hillman3 | Silviani Velina1,4 | Joo‐Won Nam1 Sang‐Jip Nam2 | Inho Choi4,5 | Kyung Sik Song6 | Geum Jin Kim7 | Hyukjae Choi1,4 | 1 College of Pharmacy, Yeungnam University, Gyeongsan, Gyeong‐buk, Republic of Korea | 2Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, Republic of Korea | 3Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Andalas, Kampus Limau Manis, Padang, Indonesia | 4Research, Institute of Cell Culture, Yeungnam University, Gyeongsan, Gyeong‐buk, Republic of Korea | 5Department of Medical Biotechnology, Yeungnam University, Gyeongsan, Gyeong‐buk, Republic of Korea | 6Research, Institute of Pharmaceutical Sciences, College of Pharmacy, Kyungpook National University, Daegu, Republic of Korea | 7Department of Pharmacology, School of Medicine, Dongguk University, Gyeongju, Gyeong‐buk, Republic of Korea Correspondence: Geum Jin Kim (geumjinkim@dongguk.ac.kr) | Hyukjae Choi (h5choi@yu.ac.kr) Received: 22 February 2025 | Revised: 6 June 2025 | Accepted: 14 June 2025 Funding: This study was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIT) (Grant No. NRF‐2020R1A6A1A03044512, and NRF‐2021R1A2C1010727), [Cooperative Research Program for Agriculture Science & Technology Development (PJ014208032023) funded by Rural Development Administration] and Yeungnam University research grant funded by Yeungnam University. Keywords: antimicrobial | antiquorum‐sensing | lipophilic fraction | Platycodon grandiflorum | polyacetylene ABSTRACT The root of Platycodon grandiflorum has long been used as a vegetable and traditional medicine. Although the antibacterial activity of the plant's lipophilic (hexanes and dichloromethane) fractions has been reported, the specific antibacterial compounds have not been identified. In this study, chemical analysis of the lipophilic fractions of P. grandiflorum extracts led to the discovery of five new polyacetylenes (1–5) and nine known compounds (6–14). Their structures were elucidated and confirmed based on 1D and 2D NMR data together with mass spectra. In particular, the relative and absolute configurations of 1 were elucidated by coupling constants, NOESY and J‐based configurational analysis in combination with Mosher's method. Compounds 1 and 2 demonstrated strong antifungal activity against Candida albicans. Additionally, compound 1 significantly inhibited quorum sensing in Chromobacterium violaceum, a commonly used biosensor strain. Compounds 2 and 14 also exhibited mild inhibitory activity. Compounds 8 and 12–14 exhibited potent antibacterial activity against Escherichia coli, Kocuria rhizophila, and Staphylococcus aureus, as well as antifungal activity against Candida albicans. These compounds may be responsible of the antimicrobial activity of P. grandiflorum. 1 | Introduction Platycodon grandiflorum is an herbaceous perennial plant in the family Campanulaceae, widely distributed in East Asia. The root of P. grandiflorum is a common vegetable and traditional medicine in Korea, China, and Japan. As a vegetable in Korean cuisine, it is called Doraji and used as raw material in cooking or for wine and tea preparation, with its characteristic flavor Hyukbean Kwon and Jimin Moon contributed equally to this study. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. © 2025 The Author(s). Archiv der Pharmazie published by Wiley‐VCH GmbH on behalf of Deutsche Pharmazeutische Gesellschaft. Archiv der Pharmazie, 2025; 358:e70043 https://doi.org/10.1002/ardp.70043 1 of 11 Escherichia coli KCTC2441 (Supporting Information S1: Table S1). Thus, the lipophilic (hexanes and dichloromethane) fractions of P. grandiflorum extracts were selected for chemical investigation, leading to the isolation of five novel polyacetylenes (1–5), along with nine known compounds (6–14), and the discovery of antimicrobial compounds among the isolates (Figure 1). Extensive inspection of 1D and 2D NMR data, together with J‐based configuration analysis for 1, was conducted to elucidate its planar structure with relative configurations. The absolute configurations of 1 were determined using advanced Mosher's analysis. Furthermore, the antibacterial, antifungal, and quorum‐sensing inhibitory activities of isolates (1–2, 5–14) were evaluated to uncover the principles of the antimicrobial activities of P. grandiflorum. 2 | Results and Discussion 2.1 | Structural Elucidation In our preliminary study, a 70% ethanolic extract of P. grandiflorum and its hexanes fraction showed antibacterial activity against Staphylococcus aureus KCTC1927 and FIGURE 1 2 of 11 Platypyran A (1) was isolated as a brown oil. Its molecular formula, C14H18O2, was determined by a protonated ion peak at | Structures of compounds (1–14) isolated from the roots of Platycodon grandiflorum. Archiv der Pharmazie, 2025 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License and bitter taste. Additionally, it is used in traditional medicine (Platycodonis radix) to treat colds, coughs, sore throats, upper respiratory infections, tonsillitis, and chest congestion in East Asia [1]. To date, many natural products from this plant have been reported, including triterpene saponins, polysaccharides [2], flavonoids [3, 4], phenolic acids [5], polyacetylenes [6–8], and sterols [9]. Triterpenoid saponins, such as platycodin D, are the major bioactive components of Platycodonis radix, with over 60 triterpene saponins reported [10]. Extracts from this plant have shown antitumor [11, 12], antitussive [1], anti‐inflammatory [13], antiobesity [14], antifibrosis [15], antioxidant [16], hepatoprotective [17], blood lipid‐lowering [18], immune‐modulatory [19], and antibacterial [20] activities. Most of these pharmacological activities are attributed to triterpene saponins and polysaccharides [10]. However, the compound responsible for the antibacterial activity of P. grandiflorum remains unknown. Previously, lipophilic extracts were reported to have antibacterial and antifungal activities [21]. TABLE 1 | (δC 79.6)/C‐9 (δC 72.7). Additionally, the vicinal 1H–1H coupling at 3JH‐2,H‐3 measured at 15.9 Hz, suggested a trans configuration for the olefin. These data resulted in the planar structure of 1 (Figure 2). The relative configuration between C‐9 and C‐10 was determined through analysis of vicinal 1H–1H coupling constants, NOESY spectra, and J‐based configuration analysis [22]. The two large vicinal 1H–1H couplings between H‐10 and H‐11b (11.1 Hz) and between H‐13b and H‐14a (11.2 Hz) indicate that these protons are all axially oriented. The relatively small vicinal 1H–1H couplings between H‐10 an H‐11a (2.1 Hz), between H‐13a and H‐14a (2.8 Hz), between H‐13a and H‐14b (2.3 Hz), and between H‐13b and H‐14b (2.3 Hz) indicated that H‐11a, H‐13a, and H‐14b are in equatorial orientations. The NOESY cross‐peak between H‐11b and H‐13b further supported their axial positioning (Figure 2 and Supporting Information S1: Figure S7). The 1H‐13C heteronuclear coupling constants were measured through HETLOC experiments (Supporting Information S1: Figure S8). The key 1H–1H vicinal coupling constant at 3 JH‐9,H‐10 was evaluated at 5.6 Hz, indicating a medium‐range coupling consistent with the equilibrium between anti‐ and gauche‐rotamers. Additionally, the two relatively small 3JH,C values between H‐9 and C‐11 (1.2 Hz) as well as between C‐8 and H‐10 (2.4 Hz), along with the two medium 2JH,C values between H‐9 and C‐10 (−1.5 Hz) as well as C‐9 and H‐10 (−2.1 Hz), suggested that C‐9/C‐10 adopt a threo form, with the two major rotamers being A‐1 and A‐3 (Figure 3). This was further supported by NOESY correlations: from H‐10 to 1 H NMR data at 600 MHz and 13C NMR data at 150 MHz of compounds 1 and 2 in CDCl3 (δ in ppm, J in Hz). Compound 1 Compound 2 No. δC δH δC δH 1 19.0 1.80 dd (6.9, 1.4) 4.5 1.95 s 2 143.5 6.29 dq (15.9, 6.9) 75.2 3 109.9 5.50 dq (15.9, 1.4) 64.9 4 74.5 59.6 5 76.9 a 60.9 6 67.0 67.3 7 79.6 8 24.4 75.1 2.53 dd (17.3, 5.6) 24.1 2.62 dd (17.3, 5.6) 2.59 dd (17.4, 5.6) 9 72.7 3.58 q (5.6) 72.4 3.58 p (5.6) 10 78.7 3.36 ddd (11.1, 5.6, 2.1) 78.5 3.34 ddd (11.1, 5.6, 2.1) 11 27.8 1.44 m 27.6 1.45 m 1.57 m b 1.56 m 12 26.0 1.53 m 25.9 1.53 mb 13 23.1 1.53 mb 22.9 1.53 mb 1.87 m 14 68.7 3.46 td (11.2, 2.8) 4.01 dt (11.2, 2.3) 9‐OH a b 2.50 dd (17.4, 5.6) 1.88 m 68.6 3.46 td (11.1, 2.7) 4.01 dt (11.1, 2.2) 2.60 brsb Assigned by HMBC. Overlapped signals. 3 of 11 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License m/z 219.1391 [M + H]+ in the HR–FAB–MS spectrum, indicating six degrees of unsaturation (Supporting Information S1: Figure S1). The analysis of 1H and 13C NMR, along with phase‐ edited HSQC spectra, allowed for the assignment of H–C connectivity and the protonation of each carbon. The data revealed two olefinic methines (δC 143.5/δH 6.29; δC 109.9/δH 5.50), two oxygenated methanetriyl group (δC 78.7/δH 3.36; δC 72.7/δH 3.58), one oxygenated methylene (δC 68.7/δH 3.46, 4.01), four nonoxygenated methylenes (δC 27.8/δH 1.44, 1.57; δC 26.0/δH 1.53; δC 24.4/δH 2.53, 2.62; δC 23.1/δH 1.53, 1.87), a methyl group (δC 19.0/δH 1.80), and four nonprotonated carbons (δC 79.6, 76.9, 74.5, and 67.0) (Table 1). Analysis of 1D and 2D NMR (HSQC, COSY and HMBC) data, enabled the construction of partial structures (Supporting Information S1: Figures S2–S6). COSY correlations of H‐2 (δH 6.29) with H‐1 (δH 1.80), and H‐3 (δH 5.50) suggested a propene fragment, supported by HMBC correlations from H‐1 to C‐2 (δC 143.5) and C‐3 (δC 109.9). A series of COSY correlations from H‐8 to H‐14 via H‐9, H‐10, H‐11, H‐12, and H‐13, along with HMBC correlations from H‐14 (δH 3.46 and 4.01) to C‐10 (δC 78.7), suggested the presence of a 1‐(tetrahydro‐2H‐pyran‐2‐yl)ethanol spin system. The two partial structures were connected via a diyne linkage based on the long‐range HMBC correlations from H‐1 to C‐4 (δC 74.5)/C‐6 (δC 67.0)/C‐7 (δC 79.6)/C‐9 (δC 72.7), from H‐2 (δH 6.29) to C‐4 (δC 74.5)/C‐6 (δC 67.0), from H‐3 (δH 5.50) to C‐4 (δC 74.5)/C‐6 (δC 67.0)/C‐7 (δC 79.6)/C‐9 (δC 72.7), from H‐8a (δH 2.53) to C‐2 (δC 143.5)/C‐3 (δC 109.9)/C‐4 (δC 74.5)/C‐6 (δC 67.0)/C‐7 (δC 79.6)/C‐9 (δC 72.7), and from H‐8b (δH 2.62) to C‐2 (δC 143.5)/C‐3 (δC 109.9)/C‐4 (δC 74.5)/C‐6 (δC 67.0)/C‐7 H‐8a/H‐8b/H‐9 and from H‐9 to H‐11a/H‐11b for the A‐1 rotamer; from H‐8 to H‐10/H‐11b, and from H‐9 to H‐11a/H‐11b for the A‐3 rotamer (Figure 3). Comparison of 1H NMR data with previously reported synthetic model compounds showed strong agreement with the threo configuration [23]. The absolute configuration of compound 1 was determined using the advanced Mosher's method, and the ΔδS−R values indicated the R configurations for both C‐9 and C‐10 (Figure 4) [24]. Consequently, the absolute configuration of compound 1 was unambiguously established as 2E, 9R, and 10R. 4 of 11 Platypyran B (2) was isolated as yellow oil. The molecular formula of 2 was determined as C14H16O2 by an ion peak of protonated molecule at m/z 217.1228 [M+H]+ in the HR–FAB–MS spectrum, indicating 7 degrees of unsaturation in 2 (Supporting Information S1: Figure S9). The similarities in the 1 H NMR spectrum, along with mass spectrometry data, suggested a structural resemblance between compounds 2 and 1 (Table 1). The distinctive differences in the 1H and 13C NMR spectra of 2 from those of 1 were the absence of signals from a double bond and the presence of two additional unprotonated Archiv der Pharmazie, 2025 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License FIGURE 2 | Key 2D NMR correlations of 1. (A) Planar structure of 1 with key COSY and HMBC correlations. (B) Key NOESY correlations. (C) 1H–1H coupling constants based on proton assignment of axial‐equatorial orientation on a pyran ring structure of 1. (D) Zoomed in 1H NMR spectra of H‐9, H‐10, H‐14a, and H‐14b. correlations [8]. However, similarities in the 1H and 13C NMR spectra including chemical shifts and 1H–1H coupling constants of isolobetyol with compounds 1 and 2 suggested the relative configuration of 9R* and 10R* to be threo form. FIGURE 4 | ΔδS–R values of the MTPA esters of 1 in pyridine‐d5. carbons at δC 75.2 and δC 64.9. Extensive 1D and 2D NMR data analysis (Supporting Information S1: Figures S10–S14) suggested that compound 2 had a triyne structure, as shown in Figure 1. The key HMBC correlations from H‐1 (δH 1.95) to C‐2 (δC 75.2)/C‐3 (δC 64.9)/C‐4 (δC 59.6)/C‐6 (δC 67.3), and from H‐8 (δH 2.50 and 2.59) to C‐4 (δC 59.6)/C‐6 (δC 67.3)/C‐7 (δC 75.1) strongly supported the connection between the methyl group and partial structures via a triyne linkage. A careful comparison of 1H NMR spectra of 2 with that of 1 revealed an addition of exchangeable proton signal at δH 2.60, assigned to 9‐OH, and a difference in H‐9 multiplicity (a pentet in 2 vs. a quartet in 1) caused by additional 1H–1H coupling between H‐9 and 9‐OH in 2. The relative configuration of 2 was inferred based on the high similarity of 1 and 2 in their 1H and 13C NMR chemical shifts and 1H–1H coupling constants. The specific rotation of 2 was measured as [α]D30 − 3.71 (c 0.1, CH2Cl2), very similar to that of 1 {[α]D30 − 3.17 (c 0.1, CH2Cl2)}. Thus, the absolute configurations of 2 were assigned as 9R and 10R. Previously, isolobetyol, a tetrahydropyran‐bearing polyacetylene from P. grandiflorum, was reported with the erythro configuration for its 1‐(tetrahydro‐2H‐pyran‐2‐yl)ethanol unit, based on NOESY Pilosulinene D (3) was purified as a white powder. The molecular formula of compound 3 was determined as C16H18O3 by HR–ESI–MS at m/z 257.1187 [M–H]–, indicating seven degrees of unsaturation (Supporting Information S1: Figure S15). The 1H NMR spectrum showed signals for a methyl proton (δH 1.77), an oxygenated methine (δH 5.36), three aromatic protons (δH 7.24, 7.17, 6.98), a hydroxy proton (δH 5.65), and four olefinic protons (δH 6.89, 6.09, 6.07, 5.55). The 13C NMR spectrum displayed signals for a methyl carbon (δC 19.0), a di‐oxygenated methine (δC 101.8), ten sp2 carbons (δC 153.0, 141.9, 136.9, 134.0, 129.0, 121.9, 119.0, 117.2, 113.1, 110.4), and two unprotonated carbons (δC 97.1, 83.9) (Table 2 and Supporting Information S1: Figures S16 and S17). COSY spectrum showed the correlations of H‐3 (δH 7.24) with H‐2 (δH 7.17) and H‐4 (δH 6.98), H‐7 (δH 6.89) and H‐8 (δH 6.07), as well as H‐12 (δH 6.09) with H‐11 (δH 5.55) and H‐13 (δH 1.77) (Supporting Information S1: Figure S18). HMBC correlations from H‐2 (δH 7.17) to C‐6 (δC 121.9), from H‐3 (δH 7.24) to C‐1 (δC 136.9)/C‐5 (δC 153.0), from H‐4 (δH 6.98) to C‐5 (δC 153.0)/ C‐6 (δC 121.9), from H‐7 (δH 6.89) to C‐1 (δC 136.9)/C‐5 (δC 153.0)/C‐9 (δC 83.9), and from H‐13 (δH 1.77) to C‐9 (δC 83.9)/ C‐10 (δC 97.1) supported the partial structure of an alkyne‐ substituted phenol (Figure 5 and Supporting Information S1: Figures S19 and S20). Additional HMBC correlations from H‐14 (δH 5.36) to C‐2 (δC 119.0)/C‐6 (δC 121.9)/C‐15 (δC 53.1)/ C‐16 (δC 53.1) suggested a di‐O‐methylated acetal at the C‐1 position (Figure 5). The two vicinal coupling constants between H‐7 and H‐8 (3JH‐7, H‐8 = 11.4 Hz), and between H‐11 and H‐12 (3JH‐11,H‐12 = 15.7 Hz) indicated the cis configuration at H‐7/H‐8 and the trans configurations at H‐11/H‐12, respectively. Pilosulinene C (4) was also isolated as a white powder. The molecular formula of compound 4 was determined to be 5 of 11 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License FIGURE 3 | Approaches for determination of relative configurations at C9–C10 of 1. Comparison data of homo‐ and hetero‐nucleus coupling constants with J‐based configuration analysis (JBCA) models. Newman projections for C‐10/C‐9 by analysis result of JBCA. | 1 H NMR data at 600 MHz and 13C NMR data at 150 MHz of compounds 3–6 in CDCl3 (δ in ppm, J in Hz). Compound 3 δC Compound 5 δH δC 1 136.9 2 119.0 3 4 5 153.0 153.4 152.3 153.8 6 121.9 121.9 123.5 125.5 7 134.0 6.89, d (11.4) 134.0 7.07, d (16.6) 132.7 7.06, d (11.5) 132.5 7.33, d (16.4) 8 113.1 6.07, dd (11.4, 2.0) 115.7 6.26, dd (16.6, 1.9) 115.0 6.21, dd (11.5, 2.4) 117.9 6.14, dd (16.4, 1.7) 9 83.9 86.7a 93.8 86.1 10 97.1 a 96.9 92.7 11 110.4 5.55, dt (15.7, 2.0) 110.9 5.68, dt (15.7, 1.9) 110.0 5.52, dt (15.8, 1.8) 110.7 5.69, dt (15.8, 1.7) 12 141.9 6.09, dq (15.7, 6.9) 140.5 6.23, dq (15.7, 6.9) 141.9 6.10, dq (15.8, 6.9) 141.2 6.26, dq (15.8, 7.0) 13 19.0 1.77, dd (6.9, 2.0) 19.0 1.84, dd (6.9, 1.9) 18.5 1.77, dd (6.9, 1.8) 19.0 1.85, dd (7.0, 1.7) 14 101.8 5.36, s 101.6 5.38, s 192.7 10.11, s 192.1 10.15, s 15 53.1 3.30, s 53.3 3.31, s 16 53.1 3.30, s 53.3 3.31, s 137.0 δH Compound 6 δC 135.2 135.1 129.0 7.24, t (7.9) 128.6 7.17, m 129.6 7.39, t (7.8) 129.2 7.34, d (7.7) 117.2 6.98, dd (7.9, 1.1) 116.3 6.87, dd (6.1, 3.1) 122.2 7.22, dd (7.8, 1.2) 121.3 7.13, d (7.7) 5.65, brs 123.9 δH 119.4 91.5 7.17, m δC 7.17, dd (7.9, 1.1) 5‐OH a δH Compound 4 No. 5.30, brs 7.50, dd (7.8, 1.2) 5.63, brs 123.9 7.46, d (7.7) 5.44, brs Assigned by HMBC. FIGURE 5 | Planar structure of 3 based on key COSY and HMBC correlations. C16H18O3 based on the deprotonated molecular ion found in HR– ESI–MS (m/z 257.1187 [M–H]–) as shown in Supporting Information S1: Figure S21. The 1D NMR spectra of 4 are highly similar to those of 3 (Table 2 and Supporting Information S1: Figures S22–S26). The notable difference in the 1H NMR spectrum of 4 was the larger 1H–1H vicinal coupling constant between H‐7 and H‐8 (16.6 Hz in 4, 11.4 Hz in 3), indicating a trans configuration for H‐7/H‐8 in 4. The acetal groups in 3 and 4 may have been converted from aldehydes through the addition of alcohols (Supporting Information S1: Figure S27). Therefore, compounds 3 and 4 could be artifacts generated during purification. Careful inspection of the 6 of 11 LR‐LC‐MS data for the parent fraction (PG‐H2) did not reveal peaks corresponding to 3 and 4 (m/z 257 [M–H]–). However, peaks with m/z values of 211, corresponding to the respective aldehydes, were observed. This finding implied that extract or fraction was relatively less changeable than purified compound. It also supported that the potential formation of artifacts should be considered while alcohols was used in separation of aldehyde compounds. As a result, compound isolation was carried out using normal‐phase chromatography under alcohol‐ free mobile phase conditions. This led to the isolation of the original aldehyde‐containing compounds of 3 and 4, identified as 5 and 6, respectively. Spectroscopic data comparison identified compound 6 as a known polyacetylene, pilosulinene A [25]. Archiv der Pharmazie, 2025 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License TABLE 2 1 1 1 1 > 128 > 128 > 128 0.5 > 128 > 128 > 128 > 128 > 128 A. fumigatus KCTC6145 1 0.25 0.25 0.5 0.5 0.25 0.25 0.5 > 128 > 128 > 128 > 128 > 128 > 128 0.25 0.25 > 128 > 128 > 128 > 128 > 128 > 128 1 8 C. glabrata KCTC7219 64 0.25 C. albicansK KCTC 7122 8 Antifungal activity MIC (μg/mL) 1 2 5 6 > 128 > 128 > 128 > 128 > 128 > 128 K. pneumonia KCTC2690 > 128 > 128 > 128 S. typhimurium KCTC2515 > 128 > 128 > 128 > 128 > 128 > 128 8 E. coli KCTC2441 Gram (–) bacteria 4 > 128 8 > 128 S. aureus KCTC1927 8 > 128 > 128 > 128 > 128 > 128 > 128 > 128 B. subilis KCTC 1021 Gram (+) bacteria K. rhizophila KCTC1915 > 128 > 128 7 > 128 > 128 1 2 1 > 128 8 7 6 5 2 1 Antibacterial activity MIC (μg/mL) Antibacterial and antifungal activities of isolated compounds. | TABLE 3 An investigation into the antimicrobial and antifungal activities were evaluated against several strains of Gram‐positive and Gram‐ negative bacteria together with fungal strains that are related to antimicrobial susceptibility studies. The results of the investigation for P. grandiflorum extracts and fractions revealed that the 70% EtOH extract and its hexanes fraction inhibited both S. aureus KCTC 1927 and E. coli KCTC 2441 (Supporting Information S1: Table S1). Furthermore, 12 compounds (1–2, 5–14) were evaluated for antibacterial and antifungal activities, with results expressed as MIC (Table 3). Compounds 8, 12–14 displayed potent antibacterial activities against Gram‐positive K. rhizophila KCTC 1915 and S. aureus KCTC 1927, as well as Gram‐negative E. coli KCTC 2441, and antifungal activities against C. albicans KCTC 7122, C. glabrata KCTC 7219, and A. fumigatus KCTC 6145. Compounds 1–2, 5, and 7 showed moderate antimicrobial activity against S. aureus KCTC 1927, while compounds 1 and 2 exhibited antifungal activity against C. albicans KCTC 7122. Additionally, anti‐quorum sensing (QS) activity was tested with compounds 1–2, 5, 8, and 12–14 against Chromobacterium violaceum KCTC 2897, a Gram‐ negative QS reporter strain widely used in quorum sensing inhibition assays. Novel compound 1 showed the strongest anti‐QS activity between tested compounds in the qualitative screening of anti‐QS activity using the disc diffusion assay by inhibition of violacein production as indicator of QS inhibition (Figure 6). Compound 1 also showed the highest anti‐QS activity with a 39.6% inhibition of violacein production at 128 μg/mL. Compounds 2 and 14 exhibited weak inhibition (Supporting Information S1: Table S2). The antibacterial activities of 8 and 12 were previously reported [34, 35]; however, this study is the first report about their inhibitory effects on K. rhizophila KCTC 1915 and their antifungal activities. The antibacterial and antifungal activities of 13 and 14 were also revealed for the first time in this study. Collectively, compounds 1 and 2 are considered key contributors to the antimicrobial activities in P. grandiflorum. Pathogen fungi Cyclohexamide 14 13 12 11 10 9 Compounds > 128 > 128 > 128 2 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 > 128 4 1 1 1 > 128 > 128 > 128 > 128 0.25 0.5 0.5 0.5 > 128 > 128 > 128 0.25 0.25 0.5 0.5 > 128 1 1 > 128 > 128 1 > 128 > 128 > 128 > 128 > 128 11 9 2.2 | Antimicrobial and Antifungal Activities of Isolated Compounds 10 Compounds 12 Compounds 6–13 were isolated from the root of P. grandiflorum for the first time. Notably, compound 13 has primarily been reported as a synthetic additive for diesel fuel [32] and can be synthesized from butanal and butanol. However, butanol was never used during extraction, hexanes fraction preparation, or purification in this study, suggesting that compound 13 is naturally derived. > 128 Vancomycin Ampicillin 13 14 The structures of other known analogs (7–14) were confirmed by careful comparison of their MS and NMR data along with specific rotations, to literature values. They were identified as 9Z,16E‐octadeca‐9,16‐dien‐12,14‐diynoic acid (7) [26], α‐ dimorphecolic acid (8) [27], vanillin (9) [28], E−4‐(3‐ethoxyprop‐ 1‐en‐1‐yl)−2‐methoxyphenol (10) [29], butyl ferulate (11) [30], 1‐acetyl‐β‐carboline (12) [31], 1,1‐dibutoxybutane (13) [32], and lobetyol (14) [33] (Supporting Information S1: Figures S38–S73). 7 of 11 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Pilosulinene B (5) was found to have the same m/z value and a similar 1H NMR spectrum to 6. A detailed comparison of the 1H NMR spectra of 5 and 6 (Table 2) indicated that the geometry between H‐7 and H‐8 in 5 was cis (3JH‐7,H‐8 = 11.5 Hz), while in 6 it was trans (3JH‐7,H‐8 = 16.4 Hz) (Supporting Information S1: Figures S28–S37). | Anti‐QS screening results using disk diffusion assay. 3 | Conclusion As a result, five novel polyacetylenes (1–5) were isolated from the root of the edible plant, P. grandiflorum (Campanulaceae), along with nine known compounds (6–14). Among the isolated known analogs, compounds 6–13 were first observed from this plant. In addition, the absolute configurations of platypyran A (1) were unambiguously assigned using J‐based configuration analysis, 1 H–1H coupling constant analysis, and Mosher's esterification of secondary alcohols. The family Campanulaceae have been reported as one of major producer of polyacetylenes [36]. All the novel polyacetylenes in this study had a C14 skeleton, supporting the view that P. grandiflorum is a major producer of C14 polyacetylenes. Polyacetylenes were reported with multiple pharmacological activities including antibacterial activities [36] and can be considered as a phytoalexins produced by the response to tissue damage or a pathogenic infection. Therefore, polyacetylenes could be attractive molecules for further studies on characteristic structure features related to antimicrobial activities. This study identified potential antimicrobial polyacetylenes (1 and 2) through evaluation of their antimicrobial and QS inhibitory activities and demonstrated the new pharmacological potentials of the compounds 8, 12, 13, and 14. In conclusion, this study expanded the chemical space of P. grandiflorum and provided insights into its potential antimicrobial principles, furthering its medicinal use. 4 | Experimental 4.1 | Chemistry 4.1.1 | General Specific rotations were measured by a JASCO DIP‐100 polarimeter (JASCO Co., Tokyo, Japan). NMR spectra were obtained on an AVANCE NEO 600 spectrometer (NFEC‐2019‐09‐257998, Bruker 8 of 11 4.1.2 | Plant Materials The dried root of P. grandiflorum was prepared from Naemome Dah (Ulsan, Republic of Korea). The voucher specimen was identified and deposited by Dr. Hyukjae Choi at the Pharmacognosy Laboratory, Yeungnam University, Gyeongsan, Korea. 4.1.3 | Extraction and Isolation The dried roots of P. grandiflorum (9.5 kg) were extracted three times using 70% EtOH (30 L) at room temperature (RT) for 3 days. The combined extract was evaporated to yield 2.5 kg of black gummy crude extract. The dried extract (2.0 kg) was suspended in deionized water (3.5 L) and successively extracted three times with 5.0 L of hexanes, dichloromethane (DCM), ethyl acetate (EtOAc), and n‐butanol (n‐BuOH), respectively, and then concentrated to yield fractions of hexanes (PG‐H, 3.3 g), DCM (PG‐D, 25.4 g), EtOAc (PG‐E, 31.8 g), n‐BuOH (PG‐B, 154.0 g), and water (PG‐W, 2.3 kg). A portion of the hexanes fraction (3.0 g) was separated into 10 subfractions (PG‐H1–PG‐H10) by normal‐ phase (NP) MPLC with following conditions; Biotage SNAP KP‐ Sil 100 g column, a linear gradient of mixtures with DCM and MeOH from 100:0 to 0:100. PG‐H1 (13, 1018.2 mg) was further separated via MPLC. A portion of the fraction PG‐H2 (23.1 mg) was subjected to reversed‐phase (RP) preparative HPLC (Hector M 5 micron C18 100 Å, 250 × 21.2 mm) under isocratic conditions of H2O:ACN (61:39) to yield compounds 3 (0.5 mg, tR = 12.5 min) Archiv der Pharmazie, 2025 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License FIGURE 6 Switzerland AG, Fällanden, Switzerland) at the Core Research Support Center for Natural Products and Medicinal Materials (CRCNM) and on Bruker AVANCE DPX 400 and DMX 250 spectrometers at the College of Pharmacy, Yeungnam University, with CDCl3 (δC 77.16, δH 7.26) at 298 K. HR–FAB–MS spectra were recorded on a JMS‐700 mass spectrometer (JEOL, Tokyo, Japan) with a 6890 Series GC system (Agilent Technologies, Santa Clara, CA, USA). HR–ESI–MS was measured using a Q Exactive UHMR Hybrid Quadrupole‐Orbitrap MS system coupled with a Vanquish UHPLC (Thermo Fisher Scientific, Waltham, MA, USA) at the CRCNM. GCMS was recorded on a GCMS‐QP2010 (Shimadzu, Kyoto, Japan). LR–ESI–MS spectra were measured using an Agilent 6120 mass spectrometer (Santa Clara, CA, USA) with a reversed‐ phase (RP) Phenomenex Luna 3μ C18(2) 100 Å column (150 × 4.6 mm). Vacuum liquid chromatography (VLC) and open‐ column chromatography were performed using 70–230 mesh silica gel (Merck). Medium‐pressure liquid chromatography (MPLC) was performed using a Biotage Isolera One system (Uppsala, Sweden) with a Biotage SNAP KP‐Sil 100‐g silica column. HPLC isolation was conducted using a combination of a Waters 1525 binary pump/ 2487 dual‐wavelength detector or a Waters 1525 binary pump/996 Photodiode Array (PDA) detector with an analytical or semipreparative Hector M 5‐micron C18 column (250 × 4.6 or 250 × 10 mm). Preparative HPLC was performed using on a Gilson 321 HPLC system (a 321 pump and UV/Vis‐155 detector) with a preparative Hector M 5‐micron C18 100 Å column (250 × 21.2 mm). Absorbance values for the QS inhibition assay were measured using a SpectraMax ABS Plus microplate reader (Molecular Devices, San Jose, CA, USA). The InChI codes of the investigated compounds, together with some biological activity data, are provided as Supporting Information. Platypyran A (1): Brown oil; [α]D30 − 3.17 (c 0.1, CH2Cl2); HR– FAB–MS obsd. m/z 219.1391 [M+H]+, calcd. for C14H18O2+, 219.1380; 1H and 13C NMR, see Table 1. Platypyran B (2): Yellow oil; [α]D30 − 3.71 (c 0.1, CH2Cl2); HR–FAB–MS obsd. m/z 217.1228 [M+H]+, calcd. for C14H16O2+, 217.1223; 1H and 13C NMR, see Table 1. Pilosulinene B (5): White powder; HR–ESI–MS obsd. m/z 211.0759 [M–H]–, calcd. for C14H11O2–, 211.0765; 1H and 13C NMR, see Table 2. 4.1.4 | Advanced Mosher's Method Two samples of compound 1 (2 mg each) were prepared. To one vial, a catalytic amount of 4‐dimethylaminopyridine (Sigma‐ Aldrich, St. Louis, MO, USA), 20 μL of S‐(+)‐α‐methoxy‐α‐ (trifluoromethyl)phenylacetyl chloride (S‐MTPA Cl) (Sigma‐ Aldrich, St. Louis, MO, USA), and pyridine‐d5 (0.7 mL) were added. The vial was sealed and reacted for 16 h at 40°C with stirring. The other vial of 1 was derivatized with R‐MTPA Cl (Sigma‐Aldrich, St. Louis, MO, USA) following the same procedure. The resulting products were dried under a N2 stream. The evaporated materials were suspended in deionized water (1 mL) respectively, and Mosher's ester products of 1 were extracted with n‐hexane (1 mL × 3). The combined n‐hexane layers were evaporated to give the S/R‐MTPA esters of 1, and their 1H NMR spectra were measured to assign the absolute configuration of C‐9 by calculating ΔδS–R values [24]. 4.2 | Antimicrobial and Quorum‐Sensing Inhibitory Activities 4.2.1 | Minimum Inhibitory Concentration (MIC) Against Bacteria and Fungi The antibacterial and antifungal activities were tested on six bacterial strains (Bacillus subtilis KCTC1021, Escherichia coli KCTC2441, Klebsiella pneumonia KCTC 2690, Kocuria rhizophila KCTC1915, Salmonella typhimurium KCTC 2515, and Staphylococcus aureus KCTC1927) and three fungal strains (Candida albicans KCTC 7122, Candida glabrata KCTC 7219, and Aspergillus fumigatus KCTC 6145). These strains were selected as representative Gram‐positive and Gram‐negative bacteria and clinically relevant fungal pathogens, widely employed in antimicrobial susceptibility studies. These strains were inoculated in Muller–Hinton Broth (BD Difco) at 37°C for 24 h and calibrated to a McFarland standard of 0.5 (equivalent to 1.5 × 108 cfu/mL). The test samples and positive controls were prepared in DMSO at a concentration of 10 mg/mL. The stock solutions of compounds were then serially diluted twofold using Muller–Hinton Broth in 96‐well plates to a concentration range of 256–0.5 μg/mL. Then, 50 μL of bacterial cultures was added to each well, resulting in a final sample concentration range as 128–0.25 μg/mL. The total inoculum concentration was 5.0 × 105 cfu/mL. After the incubation of 96‐well plate at 37°C for 24 h, the MIC was evaluated by the observation of visible bacterial growth inhibition. Ampicillin and vancomycin served as positive controls for antibacterial activity, while cycloheximide was used as the positive control for antifungal activity [37]. Pilosulinene D (3): White powder; HR–ESI–MS obsd. m/z 257.1187 [M–H]–, calcd. for C16H18O3–, 257.1183; 1H and 13C NMR, see Table 2. 4.2.2 | Qualitative Screening of QS Inhibitory Activity Pilosulinene C (4): White powder; HR–ESI–MS obsd. m/z 257.1184 [M–H]–, calcd. for C16H18O3–, 257.1183; 1H and 13C NMR, see Table 2. A disc diffusion assay was performed to test QS inhibitory activity by measuring the inhibition of violacein pigment production of the isolates, except for compounds 3, 4, 6, 7, 9, 10, and 11 9 of 11 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License and 4 (1.0 mg, tR = 18.0 min). The remaining PG‐H2 fraction was further separated by NP analytical HPLC (Hector M Sil 5 micron, 250 × 4.6 mm) under isocratic conditions of hexanes:EtOAc (75:25), yielding fraction PG‐H2‐A (6.5 mg, tR = 19.5 min), 5 (3.9 mg, tR = 20.5 min), and 11 (2.6 mg, tR = 21.5 min). Fraction PG‐H2‐A was applied further isolation by NP analytical HPLC (Hector M Sil 5 micron, 250 × 4.6 mm) under isocratic conditions of hexanes:EtOAc (85:15) to get 6 (2.5 mg, tR = 35.2 min). Fractions PG‐H4–H6 (282.9 mg) were combined and subjected to RP semipreparative HPLC with Hector M 5 micron C18 100 Å column (250 × 10 mm) and step gradient elution of water and acetonitrile (50:50–0:100) to afford compounds 1 (32.4 mg) and 2 (5.3 mg). Subfraction PG‐H4H (1.6 mg) was further separated by RP HPLC (Hector M 5 micron C18 100 Å, 250 × 4.6 mm) to yield compound 7 (0.5 mg). PG‐H9 was chromatographed via RP semipreparative HPLC (Hector M 5 micron C18 100 Å, 250 × 10 mm) under isocratic conditions with 65% acetonitrile in deionized water, yielding compound 8 (2.0 mg). PG‐H3 (222.5 mg) was fractionated by silica gel open‐column chromatography (column size: 1.8 × 23 cm) by step gradient mixtures of hexanes and EtOAc (100:0–0:100) to yield compound 9 (9.6 mg). The second extraction of the dried roots of P. grandiflorum (9.5 kg) followed the same method as the first, and the extract was evaporated in vacuo, yielding a black gummy extract (2.3 kg). The dried extract (1.9 kg) was dissolved in water and successively extracted with hexanes, DCM, EtOAc, and n‐BuOH, and then concentrated to yield hexanes (PG2‐H, 3.6 g), DCM (PG2‐D, 20.0 g), EtOAc (PG2‐E, 12.6 g), n‐BuOH (PG2‐B, 176.2 g), and water (PG2‐ W, 2.1 kg) fractions, respectively. A portion of the hexanes layer (3.0 g) was fractionated into eight fractions (PG2‐H1–PG2‐H8) via MPLC using a silica column (Biotage SNAP KP‐Sil 100 g) following similar chromatographic methods with PG‐1 as a linear gradient of DCM:MeOH (100:0–0:100). PG2‐H3 was chromatographed via RP semipreparative HPLC with a Hector M 5 micron C18 column (250 × 10 mm) by a step gradient of acetonitrile and water (45:55–100:0), yielding compounds 10 (3.2 mg) and 12 (5.0 mg). The DCM layer (11.9 g) was fractionated into nine fractions (PG‐D1–PG‐D9) via silica gel VLC using a step gradient of hexanes and EtOAc (100:0–0:100) and EtOAc:MeOH (75:25–0:100). Fraction PG‐D7 (600.0 mg) was separated into 12 fractions (PG‐D7A–PG‐D7L) via silica open‐column chromatography using a step gradient of DCM:MeOH (100:0–0:100). PG‐D7J (99.9 mg) was purified by RP preparative HPLC (Hector M 5 micron C18 100 Å, 250 × 21.2 mm) under isocratic conditions with 35% acetonitrile to elute compound 14 (35.9 mg). Data Availability Statement 4.2.3 | Quantitative Analysis of QS Inhibitory Activity 4. A. Inada, H. Murata, M. Somekawa, and T. Nakanishi, “Phytochemical Studies of Seeds of Medicinal Plants. II. A New Dihydroflavonol Glycoside and a New 3‐Methyl‐1‐butanol Glycoside From Seeds of Platycodon grandiflorum A. DE CANDOLLE,” Chemical and Pharmaceutical Bulletin 40 (1992): 3081–3083, https://doi.org/10.1248/cpb.40.3081. The 96‐well plate assay was employed to evaluate QS inhibitory activity of the isolates, except for compounds 3, 4, 6, 7, 9, 10, and 11. The assay was conducted using C. violaceum KCTC 2897 cultured in LB broth, along with positive and negative controls. Vanillin and quercetin were used as QS inhibition (positive) controls [39, 40]. The protocol was slightly modified from previously published studies [38]. Test samples and controls were dissolved in DMSO at a stock concentration of 10 mg/mL. The stock solutions were serially diluted twofold in liquid LB medium to achieve a concentration range of 256 to 0.5 μg/mL in the 96‐well plate. Subsequently, 50 μL of calibrated bacterial suspension was added to each well, resulting in a final concentration range of 128 to 0.25 μg/mL for the test samples. The final bacterial inoculum was adjusted to 5.0 × 10⁵ CFU/mL. Plate was incubated at 37°C for 20 h, depending on the microbial strains. After incubation, 100 μL from each well was transferred to microcentrifuge tubes and centrifuged at 13,000 rpm for 10 min to precipitate violacein. The resulting pellets were resuspended in 200 μL of 1 × PBS and subjected to sonication for 30 s to extract violacein. The lysates were then centrifuged again at 13,000 rpm for 10 min to remove cell debris. Subsequently, 100 μL of the violacein‐containing supernatant was transferred to a 96‐well plate, and the absorbance was measured at 585 nm using a microplate spectrophotometer. All experiments were conducted in three independent trials, each performed in triplicate. Acknowledgments This study was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) grants funded by the Ministry of Science and ICT (Grant No. RS‐2020‐NR049591, and RS‐ 2021‐NR‐058513), [Cooperative Research Program for Agriculture Science & Technology Development (PJ014208032023) funded by Rural Development Administration], the Korea Basic Science Institute (National Research Facilities and Equipment Center) grant funded by the Ministry of Education (Grant No. RS‐2025‐02317758) and Yeungnam University research grant funded by Yeungnam University. Conflicts of Interest The authors declare no conflicts of interest. 10 of 11 The data that supports the findings of this study are available in the supporting material of this article. References 1. L. Zhang, X. Wang, J. Zhang, D. Liu, and G. Bai, “Ethnopharmacology, Phytochemistry, Pharmacology and Product Application of Platycodon grandiflorum: A Review,” Chinese Herbal Medicines 16 (2024): 327–343, https://doi.org/10.1016/j.chmed.2024.01.005. 2. Y. Zhang, M. Sun, Y. He, et al., “Polysaccharides From Platycodon grandiflorum: A Review of Their Extraction, Structures, Modifications, and Bioactivities,” International Journal of Biological Macromolecules 271 (2024): 132617, https://doi.org/10.1016/j.ijbiomac.2024.132617. 3. T. Goto, H. Tadao Kondo, H. Tamura, K. Kawahori, and H. Hattori, “Structure of Platyconin, a Diacylated Anthocyanin Isolated From the Chinese Bell‐Flower Platycodon grandiflorum,” Tetrahedron Letters 24 (1983): 2181–2184, https://doi.org/10.1016/S0040‐4039(00)81877‐8. 5. J. Y. Lee, J. W. Yoon, C. T. Kim, and S. T. Lim, “Antioxidant Activity of Phenylpropanoid Esters Isolated and Identified From Platycodon grandiflorum A. DC,” Phytochemistry 65 (2004): 3033–3039, https://doi. org/10.1016/j.phytochem.2004.08.030. 6. B. Chen, Z. Liu, Y. Zhang, et al., “Application of High‐Speed Counter‐ Current Chromatography and HPLC to Separate and Purify of Three Polyacetylenes From Platycodon grandiflorum,” Journal of Separation Science 41 (2018): 789–796, https://doi.org/10.1002/jssc.201700767. 7. J. C. Ahn, B. Hwang, H. Tada, K. Ishimaru, K. Sasaki, and K. Shimomura, “Polyacetylenes in Hairy Roots of Platycodon grandiflorum,” Phytochemistry 42 (1996): 69–72, https://doi.org/10.1016/0031‐ 9422(95)00849‐7. 8. W. Li, “Isolobetyol, a New Polyacetylene Derivative From Platycodon grandiflorum Root,” Natural Product Research 36, no. 1 (2022): 466–469, https://doi.org/10.1080/14786419.2020.1779269. 9. L. Zhang, Y. Wang, D. Yang, et al., “Platycodon grandiflorus —An Ethnopharmacological, Phytochemical and Pharmacological Review,” Journal of Ethnopharmacology 164 (2015): 147–161, https://doi.org/10. 1016/j.jep.2015.01.052. 10. L. L. Zhang, M. Y. Huang, Y. Yang, et al., “Bioactive Platycodins From Platycodonis Radix: Phytochemistry, Pharmacological Activities, Toxicology and Pharmacokinetics,” Food Chemistry 327 (2020): 127029, https://doi.org/10.1016/j.foodchem.2020.127029. 11. Y. H. Choi, D. S. Yoo, M. R. Cha, et al., “Antiproliferative Effects of Saponins From the Roots of Platycodon grandiflorumon Cultured Human Tumor Cells,” Journal of Natural Products 73 (2010): 1863–1867, https:// doi.org/10.1021/np100496p. 12. L. Zhang, Z. H. Liu, and J. K. Tian, “Cytotoxic Triterpenoid Saponins From the Roots of Platycodon grandiflorum,” Molecules 12 (2007): 832–841, https://doi.org/10.3390/12040832. 13. K. J. Jang, H. K. Kim, M. H. Han, et al., “Anti‐Inflammatory Effects of Saponins Derived From the Roots of Platycodon grandiflorus in Lipopolysaccharide‐Stimulated BV2 Microglial Cells,” International Journal of Molecular Medicine 31 (2013): 1357–1366, https://doi.org/10. 3892/ijmm.2013.1330. 14. H. L. Kim, J. Park, H. Park, et al., “Platycodon grandiflorum A. De Candolle Ethanolic Extract Inhibits Adipogenic Regulators in 3T3‐L1 Cells and Induces Mitochondrial Biogenesis in Primary Brown Preadipocytes,” Journal of Agricultural and Food Chemistry 63 (2015): 7721–7730, https://doi.org/10.1021/acs.jafc.5b01908. Archiv der Pharmazie, 2025 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License (due to quantity limitations), with a previously described method with a minor modification [38]. The strain, C. violaceum KCTC 2897, was used as the reporter strain to assess anti‐QS. Briefly, 5 mL of an overnight culture of the reporter strain was added to 45 mL LB medium containing 0.3% agar, and the mixture was cooled to approximately 45°C. The suspension was then added to the plates and left to solidify. Sterile filter paper discs (5 mm in diameter) were evenly placed on the surface of the LB agar. The test samples and reference compounds were dissolved in DMSO at a concentration of 10 mg/mL, and 20 μL of each solution was applied to the discs. The plates were incubated at 37°C for 18 h. DMSO was used as the negative control, while kanamycin, quercetin, and vanillin served as positive controls for QS inhibition [39, 40]. The anti‐QS effect was evaluated by observing the absence of violacein pigmentation on the discs. 16. J. Y. Lee, W. I. Hwang, and S. T. Lim, “Antioxidant and Anticancer Activities of Organic Extracts From Platycodon grandiflorum A. De Candolle Roots,” Journal of Ethnopharmacology 93 (2004): 409–415, https://doi.org/10.1016/j.jep.2004.04.017. 17. T. Khanal, J. H. Choi, Y. P. Hwang, Y. C. Chung, and H. G. Jeong, “Saponins Isolated From the Root of Platycodon grandiflorum Protect Against Acute Ethanol‐Induced Hepatotoxicity in Mice,” Food and Chemical Toxicology 47 (2009): 530–535, https://doi.org/10.1016/j.fct.2008.12.009. 18. K. S. Kim, O. Ezaki, S. Ikemoto, and H. Itakura, “Effects of Platycodon grandiflorum Feeding on Serum and Liver Lipid Concentrations in Rats With Diet‐Induced Hyperlipidemia,” Journal of Nutritional Science and Vitaminology 41 (1995): 485–491, https://doi.org/10.3177/jnsv.41.485. 19. Y. Xie, H. Pan, H. Sun, and D. Li, “A Promising Balanced Th1 and Th2 Directing Immunological Adjuvant, Saponins From the Root of Platycodon grandiflorum,” Vaccine 26 (2008): 3937–3945, https://doi. org/10.1016/j.vaccine.2008.01.061. 20. J. Kim, “Antibacterial and Anti‐Inflammatory Effects of Platycodon grandiflorum Extracts,” Journal of Digital Convergence 12 (2014): 359–366, https://doi.org/10.14400/JDC.2014.12.3.359. 21. I. S. Lee, M. C. Choi, and H. Y. Moon, “Effect of Platycodon grandiflorum A. DC Extract on the Bronchus Diseases Bacteria,” Korean Society for Biotechnology and Bioengineering Journal 15 (2000): 162–166. 22. N. Matsumori, D. Kaneno, M. Murata, H. Nakamura, and K. Tachibana, “Stereochemical Determination of Acyclic Structures Based on Carbon−Proton Spin‐Coupling Constants. A Method of Configuration Analysis for Natural Products,” Journal of Organic Chemistry 64 (1999): 866–876, https://doi.org/10.1021/jo981810k. 23. Y. Ahn and T. Cohen, “A General Diastereoselective Synthesis of Spiroacetals Related to Those in Ionophores via the Reaction of Lactones With Cerium(III).Gamma. Cerioalkoxide. MAD Reverses the Diastereoselectivity of the Addition of Methylmetallics to a .beta.‐Keto Ether,” Journal of Organic Chemistry 59 (1994): 3142–3150, https://doi. org/10.1021/jo00090a036. 24. K. Fujii, Y. Ikai, H. Oka, M. Suzuki, and K. Harada, “A Nonempirical Method Using LC/MS for Determination of the Absolute Configuration of Constituent Amino Acids in a Peptide: Combination of Marfey's Method With Mass Spectrometry and Its Practical Application,” Analytical Chemistry 69 (1997): 5146–5151, https://doi.org/10. 1021/ac970289b. 25. R. Y. Wang, P. J. Su, B. Li, et al., “Two New Aromatic Derivatives From Codonopsis pilosula and Their α‐Glucosidase Inhibitory Activities,” Natural Product Research 36 (2022): 4929–4935, https://doi.org/10. 1080/14786419.2021.1912749. 26. K. Oka, F. Saito, T. Yasuhara, and A. Sugimoto, “The Allergens of Dendropanax trifidus Makino and Fatsia Japonica Decne. et Planch. and Evaluation of Cross‐Reactions With Other Plants of the Araliaceae Family,” Contact Dermatitis 40 (1999): 209–213, https://doi.org/10. 1111/j.1600‐0536.1999.tb06036.x. 30. J. Chen, L. Zhang, P. Zhao, G. Ma, Q. Li, and X. Yu, “Synthesized Alkyl Ferulates With Different Chain Lengths Inhibited the Formation of Lipid Oxidation Products in Soybean Oil During Deep Frying,” Food Chemistry 410 (2023): 135458, https://doi.org/10.1016/j.foodchem.2023.135458. 31. H. J. Shin, H. S. Lee, and D. S. Lee, “The Synergistic Antibacterial Activity of 1‐Acetyl‐Beta‐Carboline and Beta‐Lactams Against Methicillin‐ Resistant Staphylococcus aureus (MRSA),” Journal of Microbiology and Biotechnology 20 (2010): 501–505, https://doi.org/10.4014/jmb.0910.10019. 32. I. I. Yoeswono and T. Falah, “1,1‐Dibutoxybutane as a Petroleum Diesel Fuel Blending Component and Their Mixture Performance,” Journal of Energy and Power Engineering 10 (2016): 726, https://doi.org/ 10.17265/1934‐8975/2016.12.002. 33. M. Ma, M. Wu, B. Tian, et al., “Polyacetylenes With Xanthine Oxidase Inhibitory Activity From the Medicinal and Edible Fruits of Cyclocodon lancifolius (Roxburgh) Kurz,” Fitoterapia 170 (2023): 105631, https://doi.org/10.1016/j.fitote.2023.105631. 34. J. M. McRae, Q. Yang, R. J. Crawford, and E. A. Palombo, “Antibacterial Compounds From Planchonia careya Leaf Extracts,” Journal of Ethnopharmacology 116 (2008): 554–560, https://doi.org/10.1016/j.jep.2008. 01.007. 35. O. Kenny, N. P. Brunton, D. Walsh, C. M. Hewage, P. McLoughlin, and T. J. Smyth, “Characterisation of Antimicrobial Extracts From Dandelion Root (Taraxacum officinale) Using LC‐SPE‐NMR,” Phytotherapy Research 29 (2015): 526–532, https://doi.org/10.1002/ptr.5276. 36. Q. Xie and C. Wang, “Polyacetylenes in Herbal Medicine: A Comprehensive Review of Its Occurrence, Pharmacology, Toxicology, and Pharmacokinetics (2014–2021),” Phytochemistry 201 (2022): 113288, https://doi.org/10.1016/j.phytochem.2022.113288. 37. I. Wiegand, K. Hilpert, and R. E. W. Hancock, “Agar and Broth Dilution Methods to Determine the Minimal Inhibitory Concentration (MIC) of Antimicrobial Substances,” Nature Protocols 3 (2008): 163–175, https://doi.org/10.1038/nprot.2007.521. 38. N. K. Vijayan, H. Tanimu, and B. O. Sukumaran, “Comparative Study on Quorum Modulatory Effect of Selected Medicinal Plants on Chromobacterium Violaceum ATCC 12472 (MTCC 2656),” Journal of Pure and Applied Microbiology 18 (2024): 1848–1859, https://doi.org/10.22207/JPAM. 18.3.34. 39. M. Skogman, S. Kanerva, S. Manner, P. Vuorela, and A. Fallarero, “Flavones as Quorum Sensing Inhibitors Identified by a Newly Optimized Screening Platform Using Chromobacterium violaceum as Reporter Bacteria,” Molecules 21 (2016): 1211, https://doi.org/10.3390/ molecules21091211. 40. D. Deryabin and A. Tolmacheva, “Antibacterial and Anti‐Quorum Sensing Molecular Composition Derived From Quercus Cortex (Oak Bark) Extract,” Molecules 20 (2015): 17093–17108, https://doi.org/10. 3390/molecules200917093. Supporting Information Additional supporting information can be found online in the Supporting Information section. 27. S. V. Naidu, P. Gupta, and P. Kumar, “Enantioselective Syntheses of (−)‐Pinellic Acid, α‐ and β‐Dimorphecolic Acid,” Tetrahedron 63 (2007): 7624–7633, https://doi.org/10.1016/j.tet.2007.05.047. 28. S. Annen, T. Zweifel, F. Ricatto, and H. Grützmacher, “Catalytic Aerobic Dehydrogenative Coupling of Primary Alcohols and Water to Acids Promoted by a Rhodium(I) Amido N‐Heterocyclic Carbene Complex,” ChemCatChem 2 (2010): 1286–1295, https://doi.org/10.1002/ cctc.201000100. 29. F. Wang, H. M. Hua, X. Bian, Y. H. Pei, and Y. K. Jing, “Two New Aromatic Compounds From the Resin of Styrax tonkinensis (Pier.) Craib,” Journal of Asian Natural Products Research 8 (2006): 137–141, https://doi.org/10.1080/10286020500480712. 11 of 11 15214184, 2025, 7, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ardp.70043 by Ewha Womans University Library, Wiley Online Library on [07/12/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License 15. M. Y. Ji, A. Bo, M. Yang, et al., “The Pharmacological Effects and Health Benefits of Platycodon grandiflorus—A Medicine Food Homology Species,” Foods 9 (2020): 142, https://doi.org/10.3390/foods9020142.