Discovery of novel ursolic acid derivatives as effective antimicrobial agents through a ROS-mediated apoptosis mechanism

doi:10.1007/s11705-023-2361-5

Front. Chem. Sci. Eng.

2023, Vol. 17

Issue (12) : 2101-2113 https://doi.org/10.1007/s11705-023-2361-5

RESEARCH ARTICLE

Discovery of novel ursolic acid derivatives as effective antimicrobial agents through a ROS-mediated apoptosis mechanism

Yihong Yang, Siyue Ma, Ting Li, Jingjing He, Shitao Liu, Hongwu Liu, Jiaojiao Zhang, Xiang Zhou(

), Liwei Liu, Song Yang(

)

National Key Laboratory of Green Pesticide, Key Laboratory of Green Pesticide and Agricultural Bioengineering (Ministry of Education), Center for R & D of Fine Chemicals of Guizhou University, Guiyang 550025, China

Download: PDF(7748 KB) HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

Abstract

In response to the reduction of food production and economic losses caused by plant bacterial diseases, it is necessary to develop new, efficient, and green pesticides. Natural products are rich and sustainable source for the development of new pesticides due to their low toxicity, easy degradation, and eco-friendliness. In this study, we prepared three series of ursolic acid derivatives and assessed their antibacterial ability. Most target compounds exhibited outstanding antibacterial activities. Among them, the relative optimal EC₅₀ values of Xanthomonas oryzae pv. oryzae and Xanthomonas axonopodis pv. citri were 2.23 (A17) and 1.39 (A16) μg·mL^–1, respectively. The antimicrobial mechanism showed that compound A17 induced an excessive accumulation and production of reactive oxygen species in bacteria and damaged the cell membrane integrity to kill bacteria. More interestingly, the addition of low concentrations of exogenous hydrogen peroxide enhanced the antibacterial efficacy of compound A17 against Xanthomonas oryzae pv. oryzae. These entertaining results suggested that compound A17 induced an apparent apoptotic behavior in the tested bacteria. Overall, we developed the promising antimicrobial agents that destroyed the redox system of phytopathogenic bacteria, further demonstrating the unprecedented potential of ursolic acid for agricultural applications.

Keywords ursolic acid antibacterial activities reactive oxygen species apoptosis

Corresponding Author(s): Xiang Zhou,Song Yang

Online First Date: 03 November 2023 Issue Date: 30 November 2023

Cite this article:

Yihong Yang,Siyue Ma,Ting Li, et al. Discovery of novel ursolic acid derivatives as effective antimicrobial agents through a ROS-mediated apoptosis mechanism[J]. Front. Chem. Sci. Eng., 2023, 17(12): 2101-2113.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-023-2361-5
https://academic.hep.com.cn/fcse/EN/Y2023/V17/I12/2101

Fig.1 (a) Bioactive derivatives of UA; (b) compounds that contain antibacterial units; (c) design strategy for target compounds.

Scheme1 Synthetic route for target compounds A1–A22.

Scheme2 Synthetic route for title molecules B1–B10.

Scheme3 Synthetic route for target compounds C1–C6.

Tab.1 EC₅₀ values of title molecules A1–A22 against pathogens in vitro

Tab.2 EC₅₀ values of title molecules B1–B10 against pathogens in vitro

Fig.2 SAR analysis of title molecules.

Tab.3 EC₅₀ values of compounds C1–C6 against pathogens in vitro

Fig.3 Fluorescence pattern of Xoo cells after incubation with compound A17 at (a) 0 μg·mL^–1, (b) 12.5 μg·mL^–1, (c) 25 μg·mL^–1, and (d) 50 μg·mL^–1, respectively (scale bar =10 μm).

Fig.4 The effect of compound A17 on enzyme activities. (a) CAT enzyme activities triggered by different dosages of compound A17; (b) SOD enzyme activities triggered by different dosages of compound A17. Statistical analysis was conducted using the ANOVA method under the condition of equal variances assumed (p > 0.05) and equal variances not assumed (p < 0.05). Different lower-case letters indicate the control efficiencies with significant differences among different treatment groups at p < 0.05.

Fig.5 Effect of exogenous H₂O₂ on the bactericidal ability of compound A17. (a) Schematic and (b) images of plate assay after treatment with 0 μg·mL^–1 H₂O₂ or compound A17, 0.05 mmol·L^–1 H₂O₂, 12.5 μg·mL^–1 compound A17, and 12.5 μg·mL^–1 compound A17 + 0.05 mmol·L^–1 H₂O₂; (c) average bacterial colony diameter after treatment with A17 and H₂O₂. Statistical analysis was conducted using the ANOVA method under the condition of equal variances assumed (p > 0.05) and equal variances not assumed (p < 0.05). Different lower-case letters indicate the control efficiencies with significant differences among different treatment groups at p < 0.05.

Fig.6 SEM images of Xoo after incubation with various dosages of compound A17 at (a) 0 μg·mL^–1, (b) 3.125 μg·mL^–1, (c) 6.25 μg·mL^–1, (d) 12.5 μg·mL^–1, (e) 25 μg·mL^–1, and (f) 50 μg·mL^–1 (scale bar = 2 μm).

Fig.7 AO-EB staining pattern of Xoo cells triggered by various concentrations of molecule A17. A17-treated Xoo cells stained with (a–c) AO and (d–f) EB. (g–i) Superimposed images of AO and EB staining (scale bar = 10 μm).

Fig.8 Schematic illustration of the action mechanism for compound A17 on Xoo cells.

Tab.4 Control effects of compound A17 against rice bacterial blight at 200 μg·mL^–1 in vivo

Tab.5 In vivo control efficiencies (14 days after spraying) of compound A16 and TC against citrus bacterial canker at 200 μg·mL^–1

Fig.9 In vivo pot assay of compounds A17, A17-OPO, A17-OSi, BT, and TC at doses of 200 μg·mL^–1 against rice bacterial blight.

1	S M Hassan, M Jasinski, Z Leonowicz, E Jasinska, A K Maji. Plant disease identification using shallow convolutional neural network. Agronomy, 2021, 11(12): 2388–2408 https://doi.org/10.3390/agronomy11122388
2	J YangH J YeH M XiangX ZhouP Y WangS S LiuB X YangH B YangL W LiuS Yang. Photo-stimuli smart supramolecular self-assembly of azobenzene/β-cyclodextrin inclusion complex for controlling plant bacterial diseases. Advanced Functional Materials, 2023, doi: 10.1002/adfm.202303206
3	W L Xiao, N Wang, L L Yang, Y M Feng, P L Chu, J J Zhang, S S Liu, W B Shao, X Zhou, L W Liu. et al.. Exploiting natural maltol for synthesis of novel hydroxypyridone derivatives as promising anti-virulence agents in bactericides discovery. Journal of Agricultural and Food Chemistry, 2023, 71(17): 6603–6616 https://doi.org/10.1021/acs.jafc.3c00465
4	P L Chu, Y M Feng, Z Q Long, W L Xiao, J Ji, X Zhou, P Y Qi, T H Zhang, H Zhang, L W Liu. et al.. Novel benzothiazole derivatives as potential anti-quorum sensing agents for managing plant bacterial diseases: synthesis, antibacterial activity assessment, and SAR study. Journal of Agricultural and Food Chemistry, 2023, 71(17): 6525–6540 https://doi.org/10.1021/acs.jafc.2c07810
5	T Odjo, D Diagne, H Adreit, J Milazzo, H Raveloson, D Andriantsimialona, A I Kassankogno, S Ravel, Y M D Gumedzoe, I Ouedraogo. et al.. Structure of african populations of Pyricularia oryzae from rice. Phytopathology, 2021, 111(8): 1428–1437 https://doi.org/10.1094/PHYTO-05-20-0186-R
6	I Talibi, H Boubaker, E H Boudyach, A Ait Ben Aoumar. Alternative methods for the control of postharvest citrus diseases. Journal of Applied Microbiology, 2014, 117(1): 1–17 https://doi.org/10.1111/jam.12495
7	K D Le, J Kim, N H Yu, B Kim, C W Lee, J C Kim. Biological control of tomato bacterial wilt, kimchi cabbage soft rot, and red pepper bacterial leaf spot using Paenibacillus elgii JCK-5075. Frontiers in Plant Science, 2020, 11: 775 https://doi.org/10.3389/fpls.2020.00775
8	J P Zubrod, M Bundschuh, G Arts, C A Bruhl, G Imfeld, A Knabel, S Payraudeau, J J Rasmussen, J Rohr, A Scharmuller. et al.. Fungicides: an overlooked pesticide class?. Environmental Science & Technology, 2019, 53(7): 3347–3365 https://doi.org/10.1021/acs.est.8b04392
9	M Mitchell, L Thornton, M A Riley. Identifying more targeted antimicrobials active against selected bacterial phytopathogens. Journal of Applied Microbiology, 2022, 132(6): 4388–4399 https://doi.org/10.1111/jam.15531
10	S S SuH W LiuJ R ZhangP Y QiY DingL ZhangL L YangL W LiuX ZhouS Yang. Discovery and structure-activity relationship studies of novel tetrahydro-β-carboline derivatives as apoptosis initiators for treating bacterial infections. Journal of Integrative Agriculture, 2023, doi: 10.1016/j.jia.2023.05.031
11	J M Gebicki. Oxidative stress, free radicals and protein peroxides. Archives of Biochemistry and Biophysics, 2016, 595: 33–39 https://doi.org/10.1016/j.abb.2015.10.021
12	A Mourenza, J A Gil, L M Mateos, M Letek. Oxidative stress-generating antimicrobials, a novel strategy to overcome antibacterial resistance. Antioxidants, 2020, 9(5): 361 https://doi.org/10.3390/antiox9050361
13	A R Phull, B Nasir, I U Haq, S J Kim. Oxidative stress, consequences and ROS mediated cellular signaling in rheumatoid arthritis. Chemico-Biological Interactions, 2018, 281: 121–136 https://doi.org/10.1016/j.cbi.2017.12.024
14	M Cobbaut, J Van Lint. Function and regulation of protein kinase D in oxidative stress: a tale of isoforms. Oxidative Medicine and Cellular Longevity, 2018, 2018: 2138502 https://doi.org/10.1155/2018/2138502
15	L J Yan, D C Allen. Cadmium-induced kidney injury: oxidative damage as a unifying mechanism. Biomolecules, 2021, 11(11): 1575 https://doi.org/10.3390/biom11111575
16	A T Dharmaraja. Role of reactive oxygen species (ROS) in therapeutics and drug resistance in cancer and bacteria. Journal of Medicinal Chemistry, 2017, 60(8): 3221–3240 https://doi.org/10.1021/acs.jmedchem.6b01243
17	M Y Memar, R Ghotaslou, M Samiei, K Adibkia. Antimicrobial use of reactive oxygen therapy: current insights. Infection and Drug Resistance, 2018, 11: 567–576 https://doi.org/10.2147/IDR.S142397
18	H Li, X D Zhou, Y Y Huang, B Y Liao, L Cheng, B Ren. Reactive oxygen species in pathogen clearance: the killing mechanisms, the adaption response, and the side effects. Frontiers in Microbiology, 2020, 11: 622534 https://doi.org/10.3389/fmicb.2020.622534
19	A Vaishampayan, E Grohmann. Antimicrobials functioning through ROS-mediated mechanisms: current insights. Microorganisms, 2021, 10(1): 61 https://doi.org/10.3390/microorganisms10010061
20	Z Y Song, Y Wu, H J Wang, H Y Han. Synergistic antibacterial effects of curcumin modified silver nanoparticles through ROS-mediated pathways. Biomaterials Advances, 2019, 99: 255–263 https://doi.org/10.1016/j.msec.2018.12.053
21	O Loiseleur. Natural products in the discovery of agrochemicals. Chimia, 2017, 71(12): 810–822 https://doi.org/10.2533/chimia.2017.810
22	G Li, H X Lou. Strategies to diversify natural products for drug discovery. Medicinal Research Reviews, 2018, 38(4): 1255–1294 https://doi.org/10.1002/med.21474
23	C L Cantrell, F E Dayan, S O Duke. Natural products as sources for new pesticides. Journal of Natural Products, 2012, 75(6): 1231–1242 https://doi.org/10.1021/np300024u
24	P G Do Nascimento, T L Lemos, A M Bizerra, A M Arriaga, D A Ferreira, G M Santiago, R Braz-Filho, J G Costa. Antibacterial and antioxidant activities of ursolic acid and derivatives. Molecules, 2014, 19(1): 1317–1327 https://doi.org/10.3390/molecules19011317
25	J Zhao, H Y Zheng, Z G Sui, F B Jing, X H Quan, W W Zhao, G W Liu. Ursolic acid exhibits anti-inflammatory effects through blocking TLR4-MyD88 pathway mediated by autophagy. Cytokine, 2019, 123: 154726 https://doi.org/10.1016/j.cyto.2019.05.013
26	W J Li, R C Yan, Y Liu, C C He, X J Zhang, Y Lu, M W Khan, C R Xu, T Yang, G Y Xiang. Co-delivery of bmi1 small interfering RNA with ursolic acid by folate receptor-targeted cationic liposomes enhances anti-tumor activity of ursolic acid in vitro and in vivo. Drug Delivery, 2019, 26(1): 794–802 https://doi.org/10.1080/10717544.2019.1645244
27	S Habtemariam. Antioxidant and anti-inflammatory mechanisms of neuroprotection by ursolic acid: addressing brain injury, cerebral ischemia, cognition deficit, anxiety, and depression. Oxidative Medicine and Cellular Longevity, 2019, 2019: 8512048 https://doi.org/10.1155/2019/8512048
28	P Y Wang, M Xiang, M Luo, H W Liu, X Zhou, Z B Wu, L W Liu, Z Li, S Yang. Novel piperazine-tailored ursolic acid hybrids as significant antibacterial agents targeting phytopathogens Xanthomonas oryzae pv. oryzae and X. axonopodis pv. citri probably directed by activation of apoptosis. Pest Management Science, 2020, 76(8): 2746–2754 https://doi.org/10.1002/ps.5822
29	P Y Qi, T H Zhang, N Wang, Y M Feng, D Zeng, W B Shao, J Meng, L W Liu, L H Jin, H Zhang, X Zhou, S Yang. Natural products-based botanical bactericides discovery: novel abietic acid derivatives as anti-virulence agents for plant disease management. Journal of Agricultural and Food Chemistry, 2023, 71(14): 5463–5475 https://doi.org/10.1021/acs.jafc.2c08392
30	J Ji, W B Shao, P L Chu, H M Xiang, P Y Qi, X Zhou, P Y Wang, S Yang. 1,3,4-Oxadiazole derivatives as plant activators for controlling plant viral diseases: preparation and assessment of the effect of auxiliaries. Journal of Agricultural and Food Chemistry, 2022, 70(26): 7929–7940 https://doi.org/10.1021/acs.jafc.2c01988
31	S Jalal, T Zhang, J Deng, J Wang, T Xu, T H Zhang, C N Zhai, R Q Yuan, H M Teng, L Huang. β-Elemene isopropanolamine derivative LXX-8250 induces apoptosis through impairing autophagic flux via PFKFB4 repression in melanoma cells. Frontiers in Pharmacology, 2022, 13: 900973 https://doi.org/10.3389/fphar.2022.900973
32	X Huang, H W Liu, Z Q Long, Z X Li, J J Zhu, P Y Wang, P Y Qi, L W Liu, S Yang. Rational optimization of 1,2,3-triazole-tailored carbazoles as prospective antibacterial alternatives with significant in vivo control efficiency and unique mode of action. Journal of Agricultural and Food Chemistry, 2021, 69(16): 4615–4627 https://doi.org/10.1021/acs.jafc.1c00707
33	W B Shao, P Y Wang, Z M Fang, J J Wang, D X Guo, J Ji, X Zhou, P Y Qi, L W Liu, S Yang. Synthesis and biological evaluation of 1,2,4-triazole thioethers as both potential virulence factor inhibitors against plant bacterial diseases and agricultural antiviral agents against tobacco mosaic virus infections. Journal of Agricultural and Food Chemistry, 2021, 69(50): 15108–15122 https://doi.org/10.1021/acs.jafc.1c05202
34	I Steiner, N Stojanovic, A Bolje, A Brozovic, D Polancec, A Ambriovic-Ristov, M R Stojkovic, I Piantanida, D Eljuga, J Kosmrlj. et al.. Discovery of ‘click’ 1,2,3-triazolium salts as potential anticancer drugs. Radiology and Oncology, 2016, 50(3): 280–288 https://doi.org/10.1515/raon-2016-0027
35	S C Jiang, X Tang, M Chen, J He, S J Su, L W Liu, M He, W Xue. Design, synthesis and antibacterial activities against Xanthomonas oryzae pv. oryzae, Xanthomonas axonopodis pv. citri and Ralstonia solanacearum of novel myricetin derivatives containing sulfonamide moiety. Pest Management Science, 2020, 76(3): 853–860 https://doi.org/10.1002/ps.5587
36	F C He, J Shi, Y J Wang, S B Wang, J X Chen, X H Gan, B A Song, D Y Hu. Synthesis, antiviral activity, and mechanisms of purine nucleoside derivatives containing a sulfonamide moiety. Journal of Agricultural and Food Chemistry, 2019, 67(31): 8459–8467 https://doi.org/10.1021/acs.jafc.9b02681
37	K J Okolotowicz, M Dwyer, D Ryan, J Cheng, E A Cashman, S Moore, M Mercola, J R Cashman. Novel tertiary sulfonamides as potent anti-cancer agents. Bioorganic & Medicinal Chemistry, 2018, 26(15): 4441–4451 https://doi.org/10.1016/j.bmc.2018.07.042
38	Y L Zhao, X Huang, L W Liu, P Y Wang, Q S Long, Q Q Tao, Z Li, S Yang. Identification of racemic and chiral carbazole derivatives containing an isopropanolamine linker as prospective surrogates against plant pathogenic bacteria: in vitro and in vivo assays and quantitative proteomics. Journal of Agricultural and Food Chemistry, 2019, 67(26): 7512–7525 https://doi.org/10.1021/acs.jafc.9b02036
39	Q Q Tao, L W Liu, P Y Wang, Q S Long, Y L Zhao, L H Jin, W M Xu, Y Chen, Z Li, S Yang. Synthesis and in vitro and in vivo biological activity evaluation and quantitative proteome profiling of oxadiazoles bearing flexible heterocyclic patterns. Journal of Agricultural and Food Chemistry, 2019, 67(27): 7626–7639 https://doi.org/10.1021/acs.jafc.9b02734
40	J J Zhang, Y M Feng, J R Zhang, W L Xiao, S S Liu, X Zhou, H Zhang, P Y Wang, L W Liu, S Yang. Resistance-driven innovations in the discovery of bactericides: novel triclosan derivatives decorating isopropanolamine moiety as promising anti-biofilm agents against destructive plant bacterial diseases. Pest Management Science, 2023, 79(7): 2443–2455 https://doi.org/10.1002/ps.7419
41	S Xu, X Z Zhao, F Liu, Y Cao, B Wang, X Y Wang, M Yin, Q Z Wang, X Feng. Crucial role of oxidative stress in bactericidal effect of parthenolide against Xanthomonas oryzae pv. oryzae. Pest Management Science, 2018, 74(12): 2716–2723 https://doi.org/10.1002/ps.5091
42	X C Yang, P L Zhang, K V Kumar, S Li, R X Geng, C H Zhou. Discovery of unique thiazolidinone-conjugated coumarins as novel broad spectrum antibacterial agents. European Journal of Medicinal Chemistry, 2022, 232: 114192 https://doi.org/10.1016/j.ejmech.2022.114192
43	W Dong, H P Liu, S Sun, Y B Wang, J L Wang. Precise engineering of dual drug-loaded polymeric nanoparticles system to improve the treatment of glioma-specific targeting therapy. Process Biochemistry, 2021, 102: 341–348 https://doi.org/10.1016/j.procbio.2021.01.018
44	H O B Oloyede, H O Ajiboye, M O Salawu, T O Ajiboye. Influence of oxidative stress on the antibacterial activity of betulin, betulinic acid and ursolic acid. Microbial Pathogenesis, 2017, 111: 338–344 https://doi.org/10.1016/j.micpath.2017.08.012
45	E Khusnutdinova, A Petrova, Z Zileeva, U Kuzmina, L Zainullina, Y Vakhitova, D Babkov, O Kazakova. Novel A-ring chalcone derivatives of oleanolic and ursolic amides with anti-proliferative effect mediated through ROS-triggered apoptosis. International Journal of Molecular Sciences, 2021, 22(18): 9796 https://doi.org/10.3390/ijms22189796
46	C Ransy, C Vaz, A Lombes, F Bouillaud. Use of H2O2 to cause oxidative stress, the catalase issue. International Journal of Molecular Sciences, 2020, 21(23): 9149 https://doi.org/10.3390/ijms21239149
47	L Han, X Y Xia, X Xiang, F H Huang, Z Zhang. Protective effects of canolol against hydrogen peroxide-induced oxidative stress in AGS cells. RSC Advances, 2017, 7(68): 42826–42832 https://doi.org/10.1039/C7RA08524A
48	B E Maleeff, T L Gales, P K Narayanan, M A Tirmenstein, T K Hart. Microscopic analysis of oxidative stress in cultured cells. I. Confocal Microscopy. Microscopy and Microanalysis, 2001, 7(S2): 634–635 https://doi.org/10.1017/S143192760002924X
49	P Belenky, J D Ye, C B Porter, N R Cohen, M A Lobritz, T Ferrante, S Jain, B J Korry, E G Schwarz, G C Walker. et al.. Bactericidal antibiotics induce toxic metabolic perturbations that lead to cellular damage. Cell Reports, 2015, 13(5): 968–980 https://doi.org/10.1016/j.celrep.2015.09.059
50	F F Li, W H Zhao, V K R Tangadanchu, J P Meng, C H Zhou. Discovery of novel phenylhydrazone-based oxindole-thiolazoles as potent antibacterial agents toward Pseudomonas aeruginosa. European Journal of Medicinal Chemistry, 2022, 239: 114521 https://doi.org/10.1016/j.ejmech.2022.114521
51	M Zhang, Y C Li, Y Bi, T L Wang, Y P Dong, Q Yang, T T Zhang. 2-Phenylethyl isothiocyanate exerts antifungal activity against Alternaria alternata by affecting membrane integrity and mycotoxin production. Toxins, 2020, 12(2): 124 https://doi.org/10.3390/toxins12020124
52	M V Shirmanova, A I Gavrina, T F Kovaleva, V V Dudenkova, E E Zelenova, V I Shcheslavskiy, A M Mozherov, L B Snopova, K A Lukyanov, E V Zagaynova. Insight into redox regulation of apoptosis in cancer cells with multiparametric live-cell microscopy. Scientific Reports, 2022, 12(1): 9058 https://doi.org/10.1038/s41598-022-12901-2
53	F Yang, C G Song, R X Ji, X Y Song, B Yang, Y Lv, Z Wei. Increasing the production of reactive oxygen species through a ferroptosis pathway disrupts the redox balance of tumor cells for cancer treatment. ACS Applied Polymer Materials, 2022, 4(7): 5001–5011 https://doi.org/10.1021/acsapm.2c00556

[1]

FCE-23039-OF-YY_suppl_1

Download

[1]	Beidou Feng, Huiyu Niu, Hongchen Zhai, Congcong Shen, Hua Zhang. In-situ hydrophobic environment triggering reactive fluorescence probe to real-time monitor mitochondrial DNA damage[J]. Front. Chem. Sci. Eng., 2022, 16(1): 92-102.

Viewed

Full text

Abstract

Cited

Shared

Discussed