New γ-pyrone derivative from Helichrysum sanguineum (L.) and its Anti-Helicobacter pylori activity

New γ-pyrone derivative from Helichrysum sanguineum (L.) and its Anti-Helicobacter pylori activity | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report New γ-pyrone derivative from Helichrysum sanguineum (L.) and its Anti- Helicobacter pylori activity Adel M Mahdi, Taha A Kedra, Muhamad Abd-Elraouf, Usama Shaheen, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7059138/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Mar, 2026 Read the published version in Revista Brasileira de Farmacognosia → Version 1 posted 6 You are reading this latest preprint version Abstract Helichrysum sanguineum is a medicinal plant traditionally used for treatment of several diseases. Phytochemical investigation of its aerial parts led to isolation of four compounds, including one new γ -pyrone derivative named Kadipyrone ( 1 ) and identified as 2-(4-hydroxy-6-methylheptan-2-yl)-3-methyl-4 H -pyran-4-one. Three known compounds were also isolated including luteolin (2) , quercetin (3) and para hydroxy benzoic acid (4) . Structures of these compounds were elucidated through extensive spectroscopic methods, including ESI-MS, HR-ESI-(qTOF)-MS, 1 H NMR, 13 C-NMR, APT-NMR experiments and two dimensional NMR techniques (HSQC and HMBC).Further, different fractions obtained from Helichrysum sanguineum and compound 1 were evaluated for their anti-bacterial activity against Helicobacter pylori through diffusion assay. The results indicated that compound 1 has a promising activity toward H. pylori with a minimum inhibitory concentration (MIC) of 32 µg/ml, suggesting the potential role of compound 1 in the treatment of H. pylori infection. Peptic ulcer Antibacterial Natural products Medicinal plants Figures Figure 1 Figure 2 Figure 3 Introduction Helicobacter pylori is one of the most pathogenic bacteria infecting humans, with approximately 50% of populations worldwide are being infected with this gram negative bacteria. Prevalence rates exceed 70% in developing countries (Hooi et al., 2017 ). H.pylori infection can lead to serious gastrointestinal complications, including peptic ulcers, duodenal ulcers, and gastric cancer (Reyes, 2023 ), as well as several extra-gastric disorders such as cardiovascular diseases, diabetes mellitus, anemia, neurologic disorders, metabolic disorders and liver diseases (Gravina et al., 2018 ). The ability of H.pylori to survive in various harsh microenvironments contributes to its significant resistance to multiple antimicrobial agents (de Souza Gonçalves et al., 2021 ). Several studies have demonstrated the efficacy of various natural products and phytochemicals in treating and eradicating of H. pylori infections (Deng et al., 2024 ). Within natural-derived products, γ-pyrone compounds are one of the most potent anti- H.pylori agents, and their activities have been investigated in previous studies (Taniguchi et al., 2000 ).Of note, some γ-pyrones have been previously isolated from Helichrysum genus (Werner et al., 2019 ). The genus Helichrysum is a large genus in the Asteraceae family and comprises approximately 600 species that are distributed worldwide. Helichrysum sanguineum (L.) Kostel, commonly known as Red Everlasting, is a perennial a perennial herbaceous species native to Mediterranean and Middle East regions, including Turkey, Lebanon, Palestine and Syria (Bahadirli et al., 2024 ).Traditional medicinal applications of H. sanguineum are well-documented across Mediterranean regions, including Turkey and Palestine, where aqueous decoction has been employed to treat asthma, tuberculosis, urogenital disorders, hepatobiliary conditions (jaundice), dermatological inflammation, diarrhea, renal calculi, and fever (Bahadirli et al., 2024 ). Although numerous studies have been conducted on Helichrysum species, several species including H. sanguineum remain poorly investigated. In this work, the chemical composition and antimicrobial activity of dichloromethane and ethyl acetate fractions of the aerial parts of H. sanguineum were investigated. Material and Methods About 2.5 kg of dried aerial parts of H. sanguineum were ere collected in April 2023 from a local Egyptian herbal market (originally imported from Syria). The plant material was authenticated by Prof. Dr. Abdo Marei, Professor of Plant Ecology, Department of Botany and Microbiology, Faculty of Science, Al-Azhar University. A voucher specimen (HS-180) was deposited in the Herbarium of the Department of Pharmacognosy and Medicinal Plants, Al-Azhar University, Cairo, Egypt. The plant material was grinded and extracted by maceration with 70% methanol (3×10L), followed by evaporation under reduced pressure to give a 253 g crude extract. The crude extract was then suspended in 500 ml distilled water and partitioned with n -Hexane (3L), dichloromethane (DCM, 3L), ethyl acetate (EtOAc, 3L) and n -butanol ( n -BuOH, 6L) to give corresponding fractions of 0.25, 3, 15 and 42 g, respectively. The antimicrobial activity of these fractions was tested against gram-positive bacterial strains including Staphylococcus epidermidis and Bacillus subtilis , gram negative bacterial strains including Helicobacter pylori , Salmonella typhimurium , and Serratia marcenscens , as well as against fungi including one filamentous fungus, Aspergillus fumigatus , and one yeast species, Candida tropicalis . All microbial strains, except H. pylori , were provided from culture collection of the Regional Center for Mycology and Biotechnology (RCMB), Al-Azhar University, Cairo, Egypt. The antimicrobial activities were evaluated using a modified well diffusion method (Rasras et al., 2023 ).Briefly, 100 µl of the test bacteria/fungi were grown on 10 ml of fresh media until they reached a count of approximately 10 8 cells/ml for bacteria or 10 5 cells/ml for fungi. One hundred µl of microbial suspension was spread onto agar plates corresponding to the broth in which they were maintained and tested for susceptibility by well diffusion method. One hundred µl of each sample (at 10 mg/ml) was added to each well (6 mm diameter holes cut in the agar gel). The plates were incubated for 24–48 h at 37°C (for bacteria and yeast) and for 48 h at 28°C (for filamentous fungi). After incubation, the microorganism's growth was observed. The resulting inhibition zone diameters were measured in millimeters and used as criterion for the antimicrobial activity. Solvent control (DMSO) was included in every experiment as negative controls.Positive controls were also performed using gentamycin as standard antibacterial drugs and ketoconazole as standard antifungal drug. The H. pylori standard strain American Type Culture Collection (ATCC 26695) was obtained from the Department of Microbiology and Immunology, Faculty of Pharmacy (Men), Al-Azhar University. H.pylori was cultured on selective media (Columbia agar with 5% sheep blood and antibiotics) under microaerophilic conditions (5–10% O₂, 5–10% CO₂, 80–90% N₂) for 3–5 days (Ma et al., 2024 ). The anti-bacterial activity of the H.sanguineum fractions and the compound 1 against H.pylori was determined at the laboratory of microbiology and immunology Department of Faculty of Pharmacy- AL -Azhar university-Cairo using Mueller-Hinton agar (MHA) assay (Mansour-Ghanaei et al., 2022 ), where A concentration of 1000 µg/ml (w/v) was prepared for each tested fraction using DMSO at concentration of 50%. Standardized concentrations (McFarland were that corresponding to 1.5X10 8 CFU) of cultures of the test bacterial isolates was prepared and swabbed aseptically on the agar plates and holes (6 mm) using a sterile cork borer. 50 µl of each fraction and controls were inoculated in each hole under aseptic condition, kept at room temperature for 1 h to allow the agents to diffuse into the agar medium and then incubated. Clarithromycin (100 µg/ml) positive control for bacteria, while DMSO (50%), used as the negative control. The MHA plates were then incubated and inverted at 37°C for 48 hours under microaerophilic condition using candle jar. The inhibition zone diameters were measured in millimeters. The MIC values were determined using broth microdilution method by polystyrene microtiter plates with 96 wells (Oy Growth Curves Ab Ltd., Helsinki, Finland), using Muller-Hinton broth as indicated by the Clinical and Laboratory Standards Institute of the United States of America (CLSI, 2020) (Huang et al., 2021 ), where a stock concentration of the tested samples was employed. Two-fold serial dilution was conducted. The wells were filled with a total volume of 200 µl, made up of 20 µl of the antimicrobial solution at a range of concentrations and 170 µl of Muller-Hinton broth and 10 µl of the tested strain at concentration of 1.5X10 8 CFU. Negative controls with 100 µl of Muller-Hinton broth and 100 µl of the tested compounds and positive controls with 200 µl of the bacterial inoculum were used. Growth was determined by adding 50 µl of Resazurin 0.01%.The DCM and EtOAc fractions exhibited the highest antimicrobial activities so they were subjected to different chromatographic techniques in order to obtain the compound/s responsible for their activities. The DCM fraction (3g) was chromatographed over a normal-phase silica gel column (2×50 cm) using DCM/MeOH (100:0 → 90:10, v/v) to give 5 sub fractions (D1-D5). Subfraction D2 (98:2 DCM/MeOH, 0.2 g) was purified over Sephadex LH-20 column (2×60 cm, 100% MeOH) to afford 25 mg of the pure compound (1) . The EtOAc fraction (15g) was chromatographed using silica gel (normal phase) column chromatography (5×50 cm) using DCM-MeOH (100%-70:30 v/v) to give 5 sub fractions (E1-E5).Sub fraction E2 (3.6g) was further chromatographed on silica gel column (2×60 cm) chromatography (normal phase) using DCM and MeOH (100:0–90:10 v/v) to give 6 sub fractions (E2a-E2f), then the sub fraction E2e (0.4g, eluted with DCM: MeOH 90:10 v/v) was purified on sephadex LH-20 column (2× 60 cm, 100% MeOH) to afford 30 mg of compound (2) .Sub fraction E3 (4.5g) was further chromatographed on silica gel column (2×60 cm) chromatography (normal phase) using DCM and MeOH (100:0–90:10 v/v) with to give 6 sub fractions (E3a-E3f), then the sub fraction E3f (0.5g, eluted with DCM: MeOH 90:10) was purified on sephadex LH-20 column (2×60 cm, 100% MeOH) to afford 60 mg of compound (3) . Sub fraction E5 (1.1g) was further chromatographed on silica gel column (2×60 cm) chromatography (normal phase) using DCM and MeOH (90:10–75:25 v/v) to give 4 sub fractions (E5a-E5d), then the sub fraction E5d (0.1g,eluted with DCM: MeOH 75:25 v/v) was purified on sephadex LH-20 column (2× 60 cm, 100% MeOH) to afford 10 mg of compound (4) . Results and Discussion Compound 1 was obtained as colorless crystalline solid; its molecular formula was determined as C 14 H 22 O 3 through HR-ESI-MS (qTOF) which showed a molecular ion peak at m/z 283.1551 [M+FA-H] - . The UV spectrum of 1 showed one absorption maximum at λ 253 nm. The 1 H NMR spectrum (Table 1) of compound 1 showed the presence of two aromatic ortho -coupled protons at δ 6.07 (d, J = 9.8 Hz, 1H) and 6.84 (d, J = 9.8 Hz, 1H), four methyl proton signals at δ 0.92 (d, J = 6.7 Hz, 3H), 0.96 (d, J = 6.6 Hz, 3H), 1.38 (s, 3H), and 1.86 (s, 3H). Additionally, the 1 H NMR spectrum revealed the presence of two methylene proton signals equivalent to four protons at δ 0.83 (dd, J = 7.4, 3.8 Hz, 2H), 1.17 (m, 2H), three methines at δ 1.23, 2.16, and 4.90. The 13 C NMR spectrum of 1 (Table 1) showed the presence of one carbonyl at δ 186.7 ppm, as well as four aromatic carbons at δ 124.7, 129.2, 159.1, and 160.2, suggesting the presence of pyrone ring. The appearance of two aromatic ortho -coupled protons in the 1 H NMR, suggested a disubstituted pyrone ring. Moreover, the 13 C NMR spectrum showed four methyl carbons resonances at δ 10.42, 21.07, 21.59, 25.63 ppm, one aliphatic oxygenated carbon at δ 67.40 ppm, two methylene carbons at δ 37.81, 49.99 ppm and two aliphatic methines at δ 28.74 and 40.02 ppm, suggesting the presence of an aliphatic hydroxylated side chain attached to the pyrone moiety. The γ- pyrone moiety was deduced from the presence of four aromatic carbons and one carbonyl carbon, in addition to the UV absorption maxima at λ 253 nm compared to that of α- pyrone at λ max 300 nm (Soliman et al., 2007). Further, the HSQC experiment was in a line with the suggestion of a γ-pyrone with an aliphatic side chain. Furthermore, the HMBC correlations (Figure 1) confirmed the structure of 1 to be γ -pyrone with a methyl group at position C3 and an aliphatic side chain at position C2. The HMBC correlations between δ H 6.07 / δ C 159.14 and 186.72, and δ H 6.8 and δ C 186.7 and 160.2, further confirmed the presence of γ-pyrone. The attachment of methyl group at C3 position of γ-pyrone ring was confirmed by correlations between the methyl protons at δ 1.86 and carbon at δ 129.23 (C-3) in HMBC spectrum. The skeleton of aliphatic side chain was confirmed through the correlations between the two terminal methyl protons at δ 0.92 and 0.96 with carbons at 28.7 and 49.99, correlation between proton signal at δ 0.83 and carbon at δ 67.40 ppm, correlation between proton signal at δ 4.9 and carbon at δ 37.8, and correlation between proton signal at δ 1.38 and carbons at δ 37.8 and 40.02. The attachment of aliphatic side chain at C-2 of the γ- pyrone ring was confirmed through the correlations of proton at δ 1.38 with carbon at 160.23 (C-2 of γ -pyrone ring). Based on the above mentioned data, compound 1 was identified as 2-(4-hydroxy-6-methylheptan-2-yl)-3-methyl- 4H -pyran-4-one and named as Kadipyrone, which is in a good agreement with related γ- pyrone derivatives in literature (Shin et al., 1986, Wilk et al., 2009, Labuda et al., 2021).This is the first report of this compound in nature. Compound 2 was obtained as light yellow powder, 1 H NMR (400 MHz, DMSO-d6) δ 6.67 (s, H3), 6.19 (d, J = 1.9 Hz, H6), 6.45 (d, J = 1.9 Hz, H8), 7.42 (d, J = 1.9 Hz, H2`), 6.89 (d, J = 8 Hz, H5`), 7.40 ppm (H-6` overlapped with H2`). 13 C NMR (100 MHz, DMSO-d6) δ 163.95 (C2), 102.86 (C3), 181.76 (C4), 161.51 (C5), 98.94 (C6), 164.39 (C7), 93.96 (C8), 157.36 (C9), 103.68 (C10), 121.47 (C1`), 113.38 (C2`), 145.85 (C3`), 149.87 (C4`), 116.11 (C5`), 119.04 (C6`). ESI/MS: m/z 285 [M-H] - (calc. for C 15 H 10 O 6 ).Compound 2 was identified as luteolin (Tawornchat et al., 2021). Compound 3 was obtained as yellow powder, 1 H NMR (400 MHz, DMSO-d6) δ 6.19 (d, J = 1.7 Hz, H6), 6.41 (d, J = 1.6 Hz, H8), 7.68 (d, J = 2.0 Hz, H2`), 6.88 (d, J = 8.5 Hz, H5`), 7.54 (dd, J = 8.5, 2.0 Hz, H6`). 13 C NMR (100 MHz, DMSO-d6) δ 147.24 (C2), 136.18 (C3), 176.29 (C4), 161.17 (C5), 98.64 (C6), 164.38 (C7), 93.81 (C8), 156.59 (C9), 103.45 (C10), 122.41 (C1`), 115.51 (C2`), 145.51 (C3`), 148.15 (C4`), 116.06 (C5`), 120.42 (C6`). ESI/MS: m/z 303 [M+H] + , 301[M+H] - (calc. for C 15 H 10 O 7 ) . Compound 3 was identified as quercetin (Sambandam et al., 2016). Compound 4 was obtained as white crystalline powder, 1 H NMR (400 MHz, DMSO-d6) δ 7.33 (d, J =8.9 2H, H2 & H6), 6.67 (d, J =8.8 2H, H3 & H5). 13 C NMR (100 MHz, DMSO-d6) δ 124.99 (C1), 130.37 (C2,C6), 115.00 (C3,C5), 160.07 (C4), 171.55 (COOH). ESI/MS m/z 139 [M+H] + (calc. for C 7 H 6 O).Compound 4 was identified as Para hydroxy benzoic acid (CHO et al., 1998). The results of antifungal assays (Table 2) showed that DCM and EtOAc fractions have good antifungal action against Candida tropicalis with MIC values of 1250 and 2500 µg/ml, respectively. Conversely, all fractions ( n -Hexane, DCM, EtOAc and n -BuOH) are inactive against Aspergillus fumigatus. Evaluation of the antibacterial activity (Table 2) showed that the DCM and EtOAc fractions exhibited good antibacterial activities against Staphylococcus epidermidis with MIC values of 625 and 1250 µg/ml, respectively, and against Salmonella typhimurium with MIC values of 2500 and 3500 µg/ml, respectively. Moreover, the results of anti H.pylori activity indicated that the DCM fraction of H. sanguineum has the highest activity with MIC value of 64 µg/ml as depicted in Table 3 and Figure 2. Importantly, compound 1, isolated from DCM fraction, was also tested against H. pylori, and the results displayed that compound 1 possess a potent antibacterial activity against H. pylori with MIC value of 32 µg/ml. In conclusion Declarations Acknowledgements The authors are grateful to Mohammed Salah Abdulrahman, Lecturer of microbiology and immunology faculty of pharmacy Alazhar University for conducting the anti-H.pylori assays. Authorship Adel M Mahdi : Plant collection, extraction, Writing – original draft, Methodology, Investigation, Formal analysis, Data curation. Taha A Kedra: original manuscript writing, data curation. Muhamad Abd-Elraouf: analysis of data, methodology, writing and revision. Usama Shaheen : analysis of data, supervision, data curation. Ezzat A. M. Genady : Writing & editing, Supervision, data curation, Conceptualization. Declaration of competing interest The authors declare no conflict of interest. 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Position δ 13 C δ 1 H ( J in Hz) HMBC 2 160.23 ………………. 3 129.23 …………………. 4 186.72 …………………. 5 124.73 6.07 (d, J = 9.8 Hz, 1H) 159.14, 186.72 6 159.14 6.84 (d, J = 9.8 Hz, 1H) 160.23, 186.72 7 40.02 2.16 (d, J = 5.0 Hz, 1H) 8 37.81 1.17 (m, 2H) 67.40 9 67.40 4.90 (m, 1H) 37.81, 49.99 10 49.99 0.83 (dd, J = 7.4, 3.8 Hz, 2H) 67.40 11 28.74 1.23 (s, 1H) 12 21.59 0.92 (d, J = 6.7 Hz, 3H) 21.07,28.74,49.99 13 10.42 1.86 (s, 3H) 129.23, 160.23, 186.72 14 25.63 1.38 (s, 3H), 37.81, 40.02,160.23 15 21.07 0.96 (d, J = 6.6 Hz, 3H) 21.59, 28.74, 49.99 J -values are reported in Hz; chemical shifts are given in ppm; assignments were confirmed by HSQC, and HMBC experiments. Data measured in dmso-d6. Table 2: Antimicrobial activity of H. sanguineum fractions (zone of inhibition and MIC). DCM EtOAc n -BuOH n -Hex Control Inhibition zone MIC Inhibition zone MIC Inhibition zone Inhibition zone Inhibition zone Aspergillus Fumigatus NA - NA - NA NA 17 Candida Tropicalis 23 1250 19 2500 15 NA 21 Staphylococcus epidermidis 22 625 15 1250 12 10 28 Bacillus subtilis 16 - 13 - 11 NA 26 Salmonella typhimurium 20 2500 15 3500 14 NA 17 Serratia marcenscens 19 - 13 - 12 NA 25 Positive control for fungi (Ketoconazole 100 µg/ml), Positive control for bacteria (Gentamycin 4 mg/ml), *NA: No activity. The sample was tested at 10 mg/ml concentration. Zone of inhibition was measured in mm. MIC was measured in µg/ml. Table 3 : Anti H-Pylori activity (well diffusion assay) of H.sanguineum DCM fraction and compound 1 and their MICs. Samples Well diffusion assay MIC (µg /ml) Plate -1 Plate-2 Plate-3 Mean SD (±) Compound 1 34 33 32 33 0.816497 32 DCM 31 30 31 30.66667 0.471405 64 Clarithromycin 24 22 23 23 0.816497 128 DMSO 50% 12 10 10 10.66667 0.942809 - Zone of inhibition is measured in mm, clarithromycin is positive control, and DMSO is negative control. MIC is expressed in (µg /ml). Supplementary Files GraphicalAbstract.docx Supportinginformations.docx Cite Share Download PDF Status: Published Journal Publication published 18 Mar, 2026 Read the published version in Revista Brasileira de Farmacognosia → Version 1 posted Reviewers agreed at journal 30 Jul, 2025 Reviewers invited by journal 30 Jul, 2025 Editor invited by journal 30 Jul, 2025 Editor assigned by journal 24 Jul, 2025 First submitted to journal 24 Jul, 2025 Editorial decision: Major revisions 15 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7059138","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":493342997,"identity":"8ce4849b-eadd-4e72-8445-191337369afe","order_by":0,"name":"Adel M Mahdi","email":"data:image/png;base64,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","orcid":"https://orcid.org/0009-0007-4223-2044","institution":"Al-Azhar University Faculty of Pharmacy for Boys","correspondingAuthor":true,"prefix":"","firstName":"Adel","middleName":"M","lastName":"Mahdi","suffix":""},{"id":493342998,"identity":"34740e50-b364-4c3d-bd0f-b02cefc35005","order_by":1,"name":"Taha A Kedra","email":"","orcid":"","institution":"Al-Azhar University Faculty of Pharmacy for Boys","correspondingAuthor":false,"prefix":"","firstName":"Taha","middleName":"A","lastName":"Kedra","suffix":""},{"id":493342999,"identity":"de179702-8402-412f-b809-17e64f2332d5","order_by":2,"name":"Muhamad Abd-Elraouf","email":"","orcid":"","institution":"Al-Azhar University Faculty of Pharmacy for Boys","correspondingAuthor":false,"prefix":"","firstName":"Muhamad","middleName":"","lastName":"Abd-Elraouf","suffix":""},{"id":493343000,"identity":"ca669198-ed49-4786-a54d-7ebb6da2076c","order_by":3,"name":"Usama Shaheen","email":"","orcid":"","institution":"Al-Azhar University Faculty of Pharmacy for Boys","correspondingAuthor":false,"prefix":"","firstName":"Usama","middleName":"","lastName":"Shaheen","suffix":""},{"id":493343001,"identity":"af7e9190-f096-4a48-9309-d925db1aa29f","order_by":4,"name":"Ezzat A. M. Genady","email":"","orcid":"","institution":"Al-Azhar University Faculty of Pharmacy for Boys","correspondingAuthor":false,"prefix":"","firstName":"Ezzat","middleName":"A. M.","lastName":"Genady","suffix":""}],"badges":[],"createdAt":"2025-07-06 17:00:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7059138/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7059138/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s43450-025-00725-w","type":"published","date":"2026-03-18T15:59:46+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":88267203,"identity":"bd875051-e868-4ab1-9b09-e7ae9a79efae","added_by":"auto","created_at":"2025-08-04 16:32:41","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32281,"visible":true,"origin":"","legend":"\u003cp\u003eKey HMBC correlations of compound\u003cstrong\u003e 1 \u003c/strong\u003e(DMSO-d6)\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/bfa0037aa1a1ffcf2d82c4e5.jpg"},{"id":88267802,"identity":"f9d75be7-913f-4c8b-9c37-6f1e90aa605e","added_by":"auto","created_at":"2025-08-04 16:40:41","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":74522,"visible":true,"origin":"","legend":"\u003cp\u003eEvaluation of Anti-H-Pylori activity (well diffusion) of DCM fraction and compound \u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN.B: HS-3 is compound \u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/12f16422df81000fe2736e3a.jpg"},{"id":88267201,"identity":"397a4d60-ed53-41d1-997c-961d252a9bc8","added_by":"auto","created_at":"2025-08-04 16:32:41","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":15916,"visible":true,"origin":"","legend":"\u003cp\u003eUnnumbered image in the Results and Discussion section.\u003c/p\u003e","description":"","filename":"Unnumberfig1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/e472c15eec96f26d1ab6e582.jpg"},{"id":105223942,"identity":"19f0cc8a-4034-43d7-b762-23bfcf75a653","added_by":"auto","created_at":"2026-03-23 16:11:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":730743,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/129dfbc9-c278-4a5e-af9e-d5f52460eef6.pdf"},{"id":88267209,"identity":"6d86d6f5-fa55-49a0-8285-4b84d7bd645d","added_by":"auto","created_at":"2025-08-04 16:32:41","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":276373,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.docx","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/4cf090baca75bb959ded149b.docx"},{"id":88267210,"identity":"46d495ba-ff9a-47ff-b4be-884738e1ab9d","added_by":"auto","created_at":"2025-08-04 16:32:41","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":4889815,"visible":true,"origin":"","legend":"","description":"","filename":"Supportinginformations.docx","url":"https://assets-eu.researchsquare.com/files/rs-7059138/v1/b5edf7c86191b93396833f3a.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003e\u003cstrong\u003eNew γ-pyrone derivative from \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHelichrysum sanguineum (L.) \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eand its Anti-\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHelicobacter pylori \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eactivity\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003e\u003cem\u003eHelicobacter pylori\u003c/em\u003e is one of the most pathogenic bacteria infecting humans, with approximately 50% of populations worldwide are being infected with this gram negative bacteria. Prevalence rates exceed 70% in developing countries (Hooi et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). \u003cem\u003eH.pylori\u003c/em\u003e infection can lead to serious gastrointestinal complications, including peptic ulcers, duodenal ulcers, and gastric cancer (Reyes, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), as well as several extra-gastric disorders such as cardiovascular diseases, diabetes mellitus, anemia, neurologic disorders, metabolic disorders and liver diseases (Gravina et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The ability of \u003cem\u003eH.pylori\u003c/em\u003e to survive in various harsh microenvironments contributes to its significant resistance to multiple antimicrobial agents (de Souza Gon\u0026ccedil;alves et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Several studies have demonstrated the efficacy of various natural products and phytochemicals in treating and eradicating of \u003cem\u003eH. pylori\u003c/em\u003e infections (Deng et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Within natural-derived products, γ-pyrone compounds are one of the most potent anti-\u003cem\u003eH.pylori\u003c/em\u003e agents, and their activities have been investigated in previous studies (Taniguchi et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).Of note, some γ-pyrones have been previously isolated from \u003cem\u003eHelichrysum\u003c/em\u003e genus (Werner et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The genus \u003cem\u003eHelichrysum\u003c/em\u003e is a large genus in the Asteraceae family and comprises approximately 600 species that are distributed worldwide. \u003cem\u003eHelichrysum sanguineum\u003c/em\u003e (L.) Kostel, commonly known as Red Everlasting, is a perennial a perennial herbaceous species native to Mediterranean and Middle East regions, including Turkey, Lebanon, Palestine and Syria (Bahadirli et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).Traditional medicinal applications of \u003cem\u003eH. sanguineum\u003c/em\u003e are well-documented across Mediterranean regions, including Turkey and Palestine, where aqueous decoction has been employed to treat asthma, tuberculosis, urogenital disorders, hepatobiliary conditions (jaundice), dermatological inflammation, diarrhea, renal calculi, and fever (Bahadirli et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Although numerous studies have been conducted on \u003cem\u003eHelichrysum\u003c/em\u003e species, several species including \u003cem\u003eH. sanguineum\u003c/em\u003e remain poorly investigated. In this work, the chemical composition and antimicrobial activity of dichloromethane and ethyl acetate fractions of the aerial parts of \u003cem\u003eH. sanguineum\u003c/em\u003e were investigated.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cp\u003eAbout 2.5 kg of dried aerial parts of \u003cem\u003eH. sanguineum\u003c/em\u003e were ere collected in April 2023 from a local Egyptian herbal market (originally imported from Syria). The plant material was authenticated by Prof. Dr. Abdo Marei, Professor of Plant Ecology, Department of Botany and Microbiology, Faculty of Science, Al-Azhar University. A voucher specimen (HS-180) was deposited in the Herbarium of the Department of Pharmacognosy and Medicinal Plants, Al-Azhar University, Cairo, Egypt. The plant material was grinded and extracted by maceration with 70% methanol (3\u0026times;10L), followed by evaporation under reduced pressure to give a 253 g crude extract. The crude extract was then suspended in 500 ml distilled water and partitioned with \u003cem\u003en\u003c/em\u003e-Hexane (3L), dichloromethane (DCM, 3L), ethyl acetate (EtOAc, 3L) and \u003cem\u003en\u003c/em\u003e-butanol (\u003cem\u003en\u003c/em\u003e-BuOH, 6L) to give corresponding fractions of 0.25, 3, 15 and 42 g, respectively.\u003c/p\u003e\u003cp\u003eThe antimicrobial activity of these fractions was tested against gram-positive bacterial strains including \u003cem\u003eStaphylococcus epidermidis\u003c/em\u003e and \u003cem\u003eBacillus subtilis\u003c/em\u003e, gram negative bacterial strains including \u003cem\u003eHelicobacter pylori\u003c/em\u003e, \u003cem\u003eSalmonella typhimurium\u003c/em\u003e, and \u003cem\u003eSerratia marcenscens\u003c/em\u003e, as well as against fungi including one filamentous fungus, \u003cem\u003eAspergillus fumigatus\u003c/em\u003e, and one yeast species, \u003cem\u003eCandida tropicalis\u003c/em\u003e. All microbial strains, except \u003cem\u003eH. pylori\u003c/em\u003e, were provided from culture collection of the Regional Center for Mycology and Biotechnology (RCMB), Al-Azhar University, Cairo, Egypt. The antimicrobial activities were evaluated using a modified well diffusion method (Rasras et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).Briefly, 100 \u0026micro;l of the test bacteria/fungi were grown on 10 ml of fresh media until they reached a count of approximately 10\u003csup\u003e8\u003c/sup\u003e cells/ml for bacteria or 10\u003csup\u003e5\u003c/sup\u003e cells/ml for fungi. One hundred \u0026micro;l of microbial suspension was spread onto agar plates corresponding to the broth in which they were maintained and tested for susceptibility by well diffusion method. One hundred \u0026micro;l of each sample (at 10 mg/ml) was added to each well (6 mm diameter holes cut in the agar gel). The plates were incubated for 24\u0026ndash;48 h at 37\u0026deg;C (for bacteria and yeast) and for 48 h at 28\u0026deg;C (for filamentous fungi). After incubation, the microorganism's growth was observed. The resulting inhibition zone diameters were measured in millimeters and used as criterion for the antimicrobial activity. Solvent control (DMSO) was included in every experiment as negative controls.Positive controls were also performed using gentamycin as standard antibacterial drugs and ketoconazole as standard antifungal drug. The \u003cem\u003eH. pylori\u003c/em\u003e standard strain American Type Culture Collection (ATCC 26695) was obtained from the Department of Microbiology and Immunology, Faculty of Pharmacy (Men), Al-Azhar University. \u003cem\u003eH.pylori\u003c/em\u003e was cultured on selective media (Columbia agar with 5% sheep blood and antibiotics) under microaerophilic conditions (5\u0026ndash;10% O₂, 5\u0026ndash;10% CO₂, 80\u0026ndash;90% N₂) for 3\u0026ndash;5 days (Ma et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The anti-bacterial activity of the \u003cem\u003eH.sanguineum\u003c/em\u003e fractions and the compound \u003cb\u003e1\u003c/b\u003e against \u003cem\u003eH.pylori\u003c/em\u003e was determined at the laboratory of microbiology and immunology Department of Faculty of Pharmacy- AL -Azhar university-Cairo using Mueller-Hinton agar (MHA) assay (Mansour-Ghanaei et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), where A concentration of 1000 \u0026micro;g/ml (w/v) was prepared for each tested fraction using DMSO at concentration of 50%. Standardized concentrations (McFarland were that corresponding to 1.5X10\u003csup\u003e8\u003c/sup\u003e CFU) of cultures of the test bacterial isolates was prepared and swabbed aseptically on the agar plates and holes (6 mm) using a sterile cork borer. 50 \u0026micro;l of each fraction and controls were inoculated in each hole under aseptic condition, kept at room temperature for 1 h to allow the agents to diffuse into the agar medium and then incubated. Clarithromycin (100 \u0026micro;g/ml) positive control for bacteria, while DMSO (50%), used as the negative control. The MHA plates were then incubated and inverted at 37\u0026deg;C for 48 hours under microaerophilic condition using candle jar. The inhibition zone diameters were measured in millimeters. The MIC values were determined using broth microdilution method by polystyrene microtiter plates with 96 wells (Oy Growth Curves Ab Ltd., Helsinki, Finland), using Muller-Hinton broth as indicated by the Clinical and Laboratory Standards Institute of the United States of America (CLSI, 2020) (Huang et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), where a stock concentration of the tested samples was employed. Two-fold serial dilution was conducted. The wells were filled with a total volume of 200 \u0026micro;l, made up of 20 \u0026micro;l of the antimicrobial solution at a range of concentrations and 170 \u0026micro;l of Muller-Hinton broth and 10 \u0026micro;l of the tested strain at concentration of 1.5X10\u003csup\u003e8\u003c/sup\u003e CFU. Negative controls with 100 \u0026micro;l of Muller-Hinton broth and 100 \u0026micro;l of the tested compounds and positive controls with 200 \u0026micro;l of the bacterial inoculum were used. Growth was determined by adding 50 \u0026micro;l of Resazurin 0.01%.The DCM and EtOAc fractions exhibited the highest antimicrobial activities so they were subjected to different chromatographic techniques in order to obtain the compound/s responsible for their activities. The DCM fraction (3g) was chromatographed over a normal-phase silica gel column (2\u0026times;50 cm) using DCM/MeOH (100:0 \u0026rarr; 90:10, v/v) to give 5 sub fractions (D1-D5). Subfraction D2 (98:2 DCM/MeOH, 0.2 g) was purified over Sephadex LH-20 column (2\u0026times;60 cm, 100% MeOH) to afford 25 mg of the pure compound \u003cb\u003e(1)\u003c/b\u003e. The EtOAc fraction (15g) was chromatographed using silica gel (normal phase) column chromatography (5\u0026times;50 cm) using DCM-MeOH (100%-70:30 v/v) to give 5 sub fractions (E1-E5).Sub fraction E2 (3.6g) was further chromatographed on silica gel column (2\u0026times;60 cm) chromatography (normal phase) using DCM and MeOH (100:0\u0026ndash;90:10 v/v) to give 6 sub fractions (E2a-E2f), then the sub fraction E2e (0.4g, eluted with DCM: MeOH 90:10 v/v) was purified on sephadex LH-20 column (2\u0026times; 60 cm, 100% MeOH) to afford 30 mg of compound \u003cb\u003e(2)\u003c/b\u003e.Sub fraction E3 (4.5g) was further chromatographed on silica gel column (2\u0026times;60 cm) chromatography (normal phase) using DCM and MeOH (100:0\u0026ndash;90:10 v/v) with to give 6 sub fractions (E3a-E3f), then the sub fraction E3f (0.5g, eluted with DCM: MeOH 90:10) was purified on sephadex LH-20 column (2\u0026times;60 cm, 100% MeOH) to afford 60 mg of compound \u003cb\u003e(3)\u003c/b\u003e. Sub fraction E5 (1.1g) was further chromatographed on silica gel column (2\u0026times;60 cm) chromatography (normal phase) using DCM and MeOH (90:10\u0026ndash;75:25 v/v) to give 4 sub fractions (E5a-E5d), then the sub fraction E5d (0.1g,eluted with DCM: MeOH 75:25 v/v) was purified on sephadex LH-20 column (2\u0026times; 60 cm, 100% MeOH) to afford 10 mg of compound \u003cb\u003e(4)\u003c/b\u003e.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eCompound \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003ewas obtained as colorless crystalline solid; its molecular formula was determined as C\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e through HR-ESI-MS (qTOF) which showed a molecular ion peak at \u003cem\u003em/z\u0026nbsp;\u003c/em\u003e283.1551 [M+FA-H]\u003csup\u003e-\u003c/sup\u003e. The UV spectrum of \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003eshowed one absorption maximum at \u0026lambda; 253 nm. The \u003csup\u003e1\u003c/sup\u003eH NMR spectrum (Table 1) of compound \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003eshowed the presence of two aromatic \u003cem\u003eortho\u003c/em\u003e-coupled protons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e6.07 (d, \u003cem\u003eJ\u0026nbsp;\u003c/em\u003e= 9.8 Hz, 1H) and 6.84 (d, \u003cem\u003eJ\u0026nbsp;\u003c/em\u003e= 9.8 Hz, 1H), four methyl proton signals at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e0.92 (d, \u003cem\u003eJ\u0026nbsp;\u003c/em\u003e= 6.7 Hz, 3H), 0.96 (d, \u003cem\u003eJ\u0026nbsp;\u003c/em\u003e= 6.6 Hz, 3H), 1.38 (s, 3H), and 1.86 (s, 3H). Additionally, \u0026nbsp;the \u003csup\u003e1\u003c/sup\u003eH NMR spectrum revealed the presence of two methylene proton signals equivalent to four protons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e0.83 (dd, \u003cem\u003eJ\u0026nbsp;\u003c/em\u003e= 7.4, 3.8 Hz, 2H), 1.17 (m, 2H), three methines at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e1.23, 2.16, and 4.90. The \u003csup\u003e13\u003c/sup\u003eC NMR spectrum of \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003e(Table 1) showed the presence of one carbonyl at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e186.7 ppm, as well as four aromatic carbons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e124.7, 129.2, 159.1, and 160.2, suggesting the presence of pyrone ring. The appearance of two aromatic \u003cem\u003eortho\u003c/em\u003e-coupled protons in the \u003csup\u003e1\u003c/sup\u003eH NMR, suggested a disubstituted pyrone ring. Moreover, the \u003csup\u003e13\u003c/sup\u003eC NMR spectrum showed four methyl carbons resonances at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e10.42, 21.07, 21.59, 25.63 ppm, one aliphatic oxygenated carbon at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e67.40 ppm, two methylene carbons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e37.81, 49.99 ppm and two aliphatic methines at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e28.74 and 40.02 ppm, suggesting the presence of an aliphatic hydroxylated side chain attached to the pyrone moiety. The \u003cem\u003e\u0026gamma;-\u003c/em\u003epyrone moiety was deduced from the presence of four aromatic carbons and one carbonyl carbon, in addition to the UV absorption maxima at \u0026lambda; 253 nm compared to that of \u003cem\u003e\u0026alpha;-\u0026nbsp;\u003c/em\u003epyrone at \u0026lambda; max 300 nm (Soliman \u003cem\u003eet al.,\u003c/em\u003e 2007). Further, the HSQC experiment was in a line with the suggestion of a \u0026gamma;-pyrone with an aliphatic side chain. Furthermore, the HMBC correlations (Figure 1) confirmed the structure of \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003eto be \u0026gamma; -pyrone with a methyl group at position C3 and an aliphatic side chain at position C2. The HMBC correlations between \u003cem\u003e\u0026delta;\u003csub\u003eH\u003c/sub\u003e\u0026nbsp;\u003c/em\u003e6.07 / \u003cem\u003e\u0026delta;\u003csub\u003eC\u003c/sub\u003e\u003c/em\u003e\u003csub\u003e\u0026nbsp;\u003c/sub\u003e159.14 and 186.72, and \u003cem\u003e\u0026delta;\u003csub\u003eH\u003c/sub\u003e\u0026nbsp;\u003c/em\u003e6.8 and \u003cem\u003e\u0026delta;\u003csub\u003eC\u003c/sub\u003e\u0026nbsp;\u003c/em\u003e186.7 and 160.2, further confirmed the presence of \u0026gamma;-pyrone. The attachment of methyl group at C3 position of \u0026gamma;-pyrone ring was confirmed by correlations between the methyl protons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e1.86 and carbon at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e129.23 (C-3) in HMBC spectrum. The skeleton of aliphatic side chain was confirmed through the correlations between the two terminal methyl protons at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e0.92 and 0.96 with carbons at 28.7 and 49.99, correlation between proton signal at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e0.83 and carbon at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e67.40 ppm, correlation between proton signal at \u003cem\u003e\u0026delta;\u003c/em\u003e 4.9 and carbon at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e37.8, and correlation between proton signal at \u003cem\u003e\u0026delta;\u0026nbsp;\u003c/em\u003e1.38 and carbons at \u003cem\u003e\u0026delta;\u003c/em\u003e 37.8 and 40.02. The attachment of aliphatic side chain at C-2 of the \u0026gamma;- pyrone ring was confirmed through the correlations of proton at \u0026delta; 1.38 with carbon at 160.23 (C-2 of \u0026gamma; -pyrone ring). Based on the above mentioned data, compound \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003ewas identified as 2-(4-hydroxy-6-methylheptan-2-yl)-3-methyl-\u003cem\u003e4H\u003c/em\u003e-pyran-4-one and named as Kadipyrone, which is in a good agreement with related \u003cem\u003e\u0026gamma;-\u003c/em\u003epyrone derivatives in literature (Shin \u003cem\u003eet al.,\u003c/em\u003e 1986, Wilk \u003cem\u003eet al.,\u003c/em\u003e 2009, Labuda \u003cem\u003eet al.,\u003c/em\u003e 2021).This is the first report of this compound in nature.\u003c/p\u003e\n\u003cp\u003eCompound \u003cstrong\u003e2\u003c/strong\u003e was obtained as light yellow powder,\u003csup\u003e\u0026nbsp;1\u003c/sup\u003eH NMR (400 MHz, DMSO-d6) \u0026delta; 6.67 (s, H3), 6.19 (d, \u003cem\u003eJ\u003c/em\u003e = 1.9 Hz, H6), 6.45 (d, \u003cem\u003eJ\u003c/em\u003e = 1.9 Hz, H8), 7.42 (d, \u003cem\u003eJ\u003c/em\u003e = 1.9 Hz, H2`), 6.89 (d, \u003cem\u003eJ\u003c/em\u003e = 8 \u0026nbsp; \u0026nbsp;Hz, H5`), 7.40 ppm (H-6` overlapped with H2`).\u003csup\u003e13\u003c/sup\u003eC NMR (100 MHz, DMSO-d6) \u0026delta; 163.95 (C2), 102.86 (C3), 181.76 (C4), 161.51 (C5), 98.94 (C6), 164.39 (C7), 93.96 (C8), 157.36 (C9), 103.68 (C10), 121.47 (C1`), 113.38 (C2`), 145.85 (C3`), 149.87 (C4`), 116.11 (C5`), 119.04 (C6`). ESI/MS: \u003cem\u003em/z\u003c/em\u003e 285 [M-H]\u003csup\u003e-\u003c/sup\u003e (calc. for C\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e).Compound \u003cstrong\u003e2\u003c/strong\u003e was identified as luteolin (Tawornchat \u003cem\u003eet al.,\u003c/em\u003e 2021).\u003c/p\u003e\n\u003cp\u003eCompound \u003cstrong\u003e3\u003c/strong\u003e was obtained as yellow powder, \u003csup\u003e1\u003c/sup\u003eH NMR (400 MHz, DMSO-d6) \u0026delta; 6.19 (d, \u003cem\u003eJ\u003c/em\u003e = 1.7 Hz, H6), 6.41 (d, \u003cem\u003eJ\u003c/em\u003e = 1.6 Hz, H8), 7.68 (d, \u003cem\u003eJ\u003c/em\u003e = 2.0 Hz, H2`), 6.88 (d, \u003cem\u003eJ\u003c/em\u003e = 8.5 Hz, H5`), 7.54 (dd, \u003cem\u003eJ\u003c/em\u003e = 8.5, 2.0 Hz, H6`).\u003csup\u003e13\u003c/sup\u003eC NMR (100 MHz, DMSO-d6) \u0026delta; 147.24 (C2), 136.18 (C3), 176.29 (C4), 161.17 (C5), 98.64 (C6), 164.38 (C7), 93.81 (C8), 156.59 (C9), 103.45 (C10), 122.41 (C1`), 115.51 (C2`), 145.51 (C3`), 148.15 (C4`), 116.06 (C5`), 120.42 (C6`). ESI/MS: \u003cem\u003em/z\u0026nbsp;\u003c/em\u003e303 [M+H]\u003csup\u003e+\u003c/sup\u003e, 301[M+H]\u003csup\u003e\u0026nbsp;-\u003c/sup\u003e (calc. for C\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e)\u003csub\u003e.\u003c/sub\u003eCompound \u003cstrong\u003e3\u003c/strong\u003e was identified as quercetin (Sambandam \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2016).\u003c/p\u003e\n\u003cp\u003eCompound \u003cstrong\u003e4\u003c/strong\u003e was obtained as white crystalline powder,\u0026nbsp;\u003csup\u003e1\u003c/sup\u003eH NMR (400 MHz, DMSO-d6) \u0026delta; 7.33 (d, \u003cem\u003eJ\u003c/em\u003e=8.9 2H, H2 \u0026amp; H6), 6.67 (d, \u003cem\u003eJ\u003c/em\u003e=8.8 2H, H3 \u0026amp; H5).\u003csup\u003e\u0026nbsp;13\u003c/sup\u003eC NMR (100 MHz, DMSO-d6) \u0026delta;\u0026nbsp;124.99 (C1), 130.37 (C2,C6), 115.00 (C3,C5), 160.07 (C4), 171.55 (COOH). ESI/MS \u003cem\u003em/z\u003c/em\u003e 139 [M+H]\u003csup\u003e+\u003c/sup\u003e (calc. for C\u003csub\u003e7\u003c/sub\u003eH\u003csub\u003e6\u003c/sub\u003eO).Compound\u0026nbsp;\u003cstrong\u003e4\u003c/strong\u003e was identified as Para hydroxy benzoic acid (CHO \u003cem\u003eet al.,\u003c/em\u003e 1998).\u003c/p\u003e\n\u003cp\u003eThe results of antifungal assays (Table 2) showed that DCM and EtOAc fractions have good antifungal action against \u003cem\u003eCandida tropicalis\u0026nbsp;\u003c/em\u003ewith MIC values of 1250 and 2500 \u0026micro;g/ml, respectively. Conversely, all fractions (\u003cem\u003en\u003c/em\u003e-Hexane, DCM, EtOAc and \u003cem\u003en\u003c/em\u003e-BuOH) are inactive against \u003cem\u003eAspergillus fumigatus.\u0026nbsp;\u003c/em\u003eEvaluation of the antibacterial activity (Table 2) showed that the DCM and EtOAc fractions exhibited good antibacterial activities against \u003cem\u003eStaphylococcus epidermidis\u0026nbsp;\u003c/em\u003ewith MIC values of 625 and 1250 \u0026micro;g/ml, respectively, and against \u003cem\u003eSalmonella typhimurium\u0026nbsp;\u003c/em\u003ewith MIC values of 2500 and 3500 \u0026micro;g/ml, respectively. Moreover, the results of anti \u003cem\u003eH.pylori\u003c/em\u003e activity indicated that the DCM fraction of \u003cem\u003eH. sanguineum\u0026nbsp;\u003c/em\u003ehas the highest activity with MIC value of 64 \u0026micro;g/ml as depicted in Table 3 and Figure 2. Importantly, compound \u003cstrong\u003e1,\u0026nbsp;\u003c/strong\u003eisolated from DCM fraction, was also tested against \u003cem\u003eH. pylori,\u0026nbsp;\u003c/em\u003eand the results displayed that compound \u003cstrong\u003e1\u0026nbsp;\u003c/strong\u003epossess a potent antibacterial activity against \u003cem\u003eH. pylori\u0026nbsp;\u003c/em\u003ewith MIC value of 32 \u0026micro;g/ml. In conclusion\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to Mohammed Salah Abdulrahman, Lecturer of microbiology and immunology faculty of pharmacy Alazhar University for conducting the \u003cem\u003eanti-H.pylori\u003c/em\u003e assays.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthorship\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdel M Mahdi\u003c/strong\u003e: Plant collection, extraction, Writing \u0026ndash; original draft, Methodology, Investigation, Formal analysis, Data curation.\u0026nbsp;\u003cstrong\u003eTaha A\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Kedra:\u003c/strong\u003e original manuscript writing, data curation. \u003cstrong\u003eMuhamad Abd-Elraouf:\u003c/strong\u003e analysis of data, methodology, writing and revision. \u003cstrong\u003eUsama Shaheen\u003c/strong\u003e: analysis of data, supervision, data curation.\u0026nbsp;\u003cstrong\u003eEzzat A. M. Genady\u003c/strong\u003e:\u0026nbsp;Writing \u0026amp; editing, Supervision, data curation, Conceptualization.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of competing interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBahadirli NP, Timur M, Sahin CB (2024) Extraction, Distribution, and Chemical Diversity of \u003cem\u003eHelichrysum plicatum, H.sanguineum and H\u003c/em\u003e.stoechas Populations from the Eastern Mediterranean Region of T\u0026uuml;rkiye. 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Bioorg Med Chem 17:2304\u0026ndash;3209. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.bmc.2008.11.001\u003c/span\u003e\u003cspan address=\"10.1016/j.bmc.2008.11.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable \u003cstrong\u003e1\u003c/strong\u003e:\u003csup\u003e\u0026nbsp;1\u003c/sup\u003eH NMR (400 MHz, DMSOd6), \u003csup\u003e13\u003c/sup\u003eC NMR (100 MHz, DMSO-d6) and HMBC of the compound \u003cstrong\u003e(1).\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"576\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003ePosition\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u0026delta;\u003csup\u003e13\u003c/sup\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e\u0026delta; \u003csup\u003e1\u003c/sup\u003eH (\u003cem\u003eJ\u003c/em\u003e in Hz)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003eHMBC\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e160.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e129.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e186.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e124.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e6.07 (d, \u003cem\u003eJ\u003c/em\u003e = 9.8 Hz, 1H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e159.14, 186.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e159.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e6.84 (d, \u003cem\u003eJ\u003c/em\u003e = 9.8 Hz, 1H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e160.23, 186.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e40.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e2.16 (d, \u003cem\u003eJ\u003c/em\u003e = 5.0 Hz, 1H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e37.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e1.17 (m, 2H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e67.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e67.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e4.90 (m, 1H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e37.81, 49.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e49.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e0.83 (dd, \u003cem\u003eJ\u003c/em\u003e = 7.4, 3.8 Hz, 2H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e67.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e28.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e1.23 (s, 1H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e21.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e0.92 (d, \u003cem\u003eJ\u003c/em\u003e = 6.7 Hz, 3H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e21.07,28.74,49.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e10.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e1.86 (s, 3H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e129.23, 160.23, 186.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e25.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e1.38 (s, 3H),\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e37.81, 40.02,160.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 11.6319%;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.5694%;\"\u003e\n \u003cp\u003e21.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.6319%;\"\u003e\n \u003cp\u003e0.96 (d, \u003cem\u003eJ\u003c/em\u003e = 6.6 Hz, 3H)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29.1667%;\"\u003e\n \u003cp\u003e21.59, 28.74, 49.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eJ\u003c/em\u003e-values are reported in Hz; chemical shifts are given in ppm; assignments were confirmed by HSQC, and HMBC experiments. Data measured in dmso-d6.\u003c/p\u003e\n\u003cp\u003eTable 2: Antimicrobial activity of\u0026nbsp;H. sanguineum\u0026nbsp;fractions (zone of inhibition and MIC).\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"690\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 20.8696%;\"\u003e\n \u003cp\u003eDCM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 22.6087%;\"\u003e\n \u003cp\u003eEtOAc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e\u003cem\u003en\u003c/em\u003e-BuOH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e\u003cem\u003en\u003c/em\u003e-Hex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eControl\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cu\u003e\u0026nbsp;\u003c/u\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eInhibition zone\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003eMIC\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eInhibition zone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eMIC\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eInhibition zone\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.913%;\"\u003e\n \u003cp\u003eInhibition zone\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eInhibition zone\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eAspergillus\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eFumigatus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eCandida\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eTropicalis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e1250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e2500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eStaphylococcus epidermidis\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e625\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e1250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eSalmonella\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003etyphimurium\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e2500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e3500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 17.3913%;\"\u003e\n \u003cp\u003e\u003cem\u003eSerratia\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003emarcenscens\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9.56522%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 11.3043%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.913%;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ePositive control for fungi (Ketoconazole 100 \u0026micro;g/ml), Positive control for bacteria (Gentamycin 4 mg/ml), *NA: No activity. The sample was tested at 10 mg/ml concentration. Zone of inhibition was measured in mm. MIC was measured in \u0026micro;g/ml.\u003c/p\u003e\n\u003cp\u003eTable \u003cstrong\u003e3\u003c/strong\u003e: Anti H-Pylori activity (well diffusion assay) of H.sanguineum DCM fraction and compound \u003cstrong\u003e1\u003c/strong\u003e and their MICs.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"621\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eSamples\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 389px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eWell diffusion assay\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eMIC (\u0026micro;g /ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 59px;\"\u003e\n \u003cp\u003ePlate -1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 63px;\"\u003e\n \u003cp\u003ePlate-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003ePlate-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 75px;\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003eSD (\u0026plusmn;)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003eCompound \u003cstrong\u003e\u0026nbsp;1\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 59px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 63px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 75px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e0.816497\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e32\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003eDCM\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 59px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 63px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 75px;\"\u003e\n \u003cp\u003e30.66667\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e0.471405\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003eClarithromycin\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 59px;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 63px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 75px;\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e0.816497\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e128\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003eDMSO 50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 59px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 63px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 75px;\"\u003e\n \u003cp\u003e10.66667\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e0.942809\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 114px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eZone of inhibition is measured in mm, clarithromycin is positive control, and DMSO is negative control. MIC is expressed in (\u0026micro;g /ml).\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"revista-brasileira-de-farmacognosia","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"rbfa","sideBox":"Learn more about [Revista Brasileira de Farmacognosia](https://www.springer.com/journal/43450)","snPcode":"43450","submissionUrl":"https://www.editorialmanager.com/rbfa/default2.aspx","title":"Revista Brasileira de Farmacognosia","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Peptic ulcer, Antibacterial, Natural products, Medicinal plants","lastPublishedDoi":"10.21203/rs.3.rs-7059138/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7059138/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eHelichrysum sanguineum\u003c/em\u003e is a medicinal plant traditionally used for treatment of several diseases.\u003c/p\u003e\n\u003cp\u003ePhytochemical investigation of its aerial parts led to isolation of four compounds, including one new \u003cstrong\u003eγ\u003c/strong\u003e -pyrone derivative named Kadipyrone (\u003cstrong\u003e1\u003c/strong\u003e) and identified as 2-(4-hydroxy-6-methylheptan-2-yl)-3-methyl-4\u003cem\u003eH\u003c/em\u003e-pyran-4-one. Three known compounds were also isolated including luteolin \u003cstrong\u003e(2)\u003c/strong\u003e, quercetin \u003cstrong\u003e(3)\u003c/strong\u003e and para hydroxy benzoic acid \u003cstrong\u003e(4)\u003c/strong\u003e. Structures of these compounds were elucidated through extensive spectroscopic methods, including ESI-MS, HR-ESI-(qTOF)-MS, \u003csup\u003e1\u003c/sup\u003eH NMR,\u003csup\u003e13\u003c/sup\u003eC-NMR, APT-NMR experiments and two dimensional NMR techniques (HSQC and HMBC).Further, different fractions obtained from \u003cem\u003eHelichrysum sanguineum\u003c/em\u003e and compound \u003cstrong\u003e1\u003c/strong\u003e were evaluated for their anti-bacterial activity against \u003cem\u003eHelicobacter pylori\u003c/em\u003e through diffusion assay. The results indicated that compound \u003cstrong\u003e1\u003c/strong\u003e has a promising activity toward \u003cem\u003eH. pylori\u003c/em\u003e with a minimum inhibitory concentration (MIC) of 32 µg/ml, suggesting the potential role of compound \u003cstrong\u003e1\u003c/strong\u003e in the treatment of \u003cem\u003eH. pylori\u003c/em\u003e infection.\u003c/p\u003e","manuscriptTitle":"New γ-pyrone derivative from Helichrysum sanguineum (L.) and its Anti-Helicobacter pylori activity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-04 16:32:36","doi":"10.21203/rs.3.rs-7059138/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-07-30T18:59:17+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-07-30T17:54:22+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Revista Brasileira de Farmacognosia","date":"2025-07-30T15:05:17+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-24T17:06:48+00:00","index":"","fulltext":""},{"type":"submitted","content":"Revista Brasileira de Farmacognosia","date":"2025-07-24T13:06:44+00:00","index":"","fulltext":""},{"type":"decision","content":"Major revisions","date":"2025-07-15T16:04:35+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"revista-brasileira-de-farmacognosia","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"rbfa","sideBox":"Learn more about [Revista Brasileira de Farmacognosia](https://www.springer.com/journal/43450)","snPcode":"43450","submissionUrl":"https://www.editorialmanager.com/rbfa/default2.aspx","title":"Revista Brasileira de Farmacognosia","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"16dcf2c1-86f3-4dee-928c-448c98451638","owner":[],"postedDate":"August 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-23T16:06:45+00:00","versionOfRecord":{"articleIdentity":"rs-7059138","link":"https://doi.org/10.1007/s43450-025-00725-w","journal":{"identity":"revista-brasileira-de-farmacognosia","isVorOnly":false,"title":"Revista Brasileira de Farmacognosia"},"publishedOn":"2026-03-18 15:59:46","publishedOnDateReadable":"March 18th, 2026"},"versionCreatedAt":"2025-08-04 16:32:36","video":"","vorDoi":"10.1007/s43450-025-00725-w","vorDoiUrl":"https://doi.org/10.1007/s43450-025-00725-w","workflowStages":[]},"version":"v1","identity":"rs-7059138","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7059138","identity":"rs-7059138","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}