Natural Polyketides Isolated from the Endophytic Fungus Phomopsis sp. CAM212 with a Semisynthetic Derivative Downregulating the ERK/IκBα Signaling Pathways

Abstract

Three previously undescribed natural products, phomopsinin A – C (1 – 3), together with three known compounds, namely, cis-hydroxymellein (4), phomoxanthone A (5) and cytochalasin L-696,474 (6), were isolated from the solid culture of Phomopsis sp. CAM212, an endophytic fungus obtained from Garcinia xanthochymus. Their structures were determined on the basis of spectroscopic data, including IR, NMR, and MS. The absolute configurations of 1 and 2 were assigned by comparing their experimental and calculated ECD spectra. Acetylation of compound 1 yielded 1a, a new natural product derivative that was tested together with other isolated compounds on lipopolysaccharide-stimulated RAW 264.7 cells. Cytochalasin L-696,474 (6) was found to significantly inhibit nitric oxide production, but was highly cytotoxic to the treated cells, whereas compound 1 slightly inhibited nitric oxide production, which was not significantly different compared to lipopolysaccharide-treated cells. Remarkably, the acetylated derivative of 1, compound 1a, significantly inhibited nitric oxide production with an IC₅₀ value of 14.8 µM and no cytotoxic effect on treated cells, thereby showing the importance of the acetyl group in the anti-inflammatory activity of 1a. The study of the mechanism of action revealed that 1a decreases the expression of inducible nitric oxide synthase, cyclooxygenase 2, and proinflammatory cytokine IL-6 without an effect on IL-1β expression. Moreover, it was found that 1a exerts its anti-inflammatory activity in lipopolysaccharide-stimulated RAW 264.7 macrophage cells by downregulating the activation of ERK1/2 and by preventing the translocation of nuclear factor κB. Thus, derivatives of phomopsinin A (1), such as compound 1a, could provide new anti-inflammatory leads.

Key words

Garcinia xanthochymus - Phomopsis sp. - polyketides - phomopsinin - anti-inflammatory - cytokines - MAPKs - NF-κB

Publikationsverlauf

Eingereicht: 29. November 2019

Angenommen nach Revision: 02. Juli 2020

Artikel online veröffentlicht:
05. August 2020

• References

• 1 Aderem A, Ulevitch RJ. Toll-like receptors in the induction of the innate immune response. Nature 2000; 406: 782-787
• 2 Hunter P. The inflammation theory of disease. The growing realization that chronic inflammation is crucial in many diseases opens new avenues for treatment. EMBO Rep 2012; 13: 968-970
• 3 Akira S, Takeda K. Toll-like receptor signalling. Nat Rev Immunol 2004; 4: 499-511
• 4 Verheij EWM, Coronel RE. Edible Fruits and Nuts. Plant Resources of South-EastAsia (PROSEA). Netherlands: Backhuys Publishers; 1991: 175
• 5 Faeth SH, Fagan WF. Fungal endophytes: common host plant symbionts but uncommon mutualists. Integr Comp Biol 2002; 42: 360-368
• 6 Kharwar RN, Mishra A, Gond SK, Stierle A, Stierle D. Anticancer compounds derived from fungal endophytes: their importance and future challenges. Nat Prod Rep 2011; 28: 1208-1228
• 7 Jouda JB, Tamokou JD, Mbazoa CD, Douala-Meli C, Sarkar P, Bag PK, Wandji J. Antibacterial and cytotoxic cytochalasins from the endophytic fungus Phomopsis sp. harbored in Garcinia kola (Heckel) nut. BMC Complement Altern Med 2016; 16: 462
• 8 Isaka M, Jaturapat A, Rukseree K, Danwisetkanjana K, Tanticharoen M, Thebtaranonth Y. Phomoxanthones A and B, novel xanthone dimers from the endophytic fungus Phomopsis species. J Nat Prod 2001; 64: 1015-1018
• 9 Hemtasin C, Kanokmedhakul S, Kanokmedhakul K, Hahnvajanawong C, Soytong K, Prabpai S, Kongsaeree P. Cytotoxic pentacyclic and tetracyclic aromatic sesquiterpenes from Phomopsis archeri. J Nat Prod 2011; 74: 609-613
• 10 Horn W, Simmonds M, Schwartz R, Blaney W. Phomopsichalasin, a novel antimicrobial agent from an endophytic Phomopsis sp. Tetrahedron 1995; 51: 969-978
• 11 Chapla VM, Zeraik ML, Ximenes VF, Zanardi LM, Lopes MN, Cavalheiro AJ, Silva DH, Young MC, Fonseca LM, Bolzani VS, Araujo AR. Bioactive secondary metabolites from Phomopsis sp., an endophytic fungus from Senna spectabilis. Molecules 2014; 19: 6597-6608
• 12 Rukachaisirikul V, Sommart U, Phongpaichit S, Sakayaroj J, Kirtikara K. Metabolites from the endophytic fungus Phomopsis sp. PSU-D15. Phytochemistry 2008; 69: 783-787
• 13 Tang JW, Wang WG, Li A, Yan BC, Chen R, Li XN, Du X, Sun HD, Pu JX. Polyketides from the endophytic fungus Phomopsis sp. sh917 by using the one strain/many compounds strategy. Tetrahedron 2017; 73: 3577-3584
• 14 Ju ZR, Lin XP, Li M, Wang Y, Tian YQ, Wang JF, Liu J, Tu Z, Xu SH, Liu Y. Chaetochromones A–C, Three New Polyketides from Mangrove Plant Derived Endophytic Fungus Phomopsis sp. SCSIO 41006. Chem Biodivers 2017; 14
• 15 Luo Y, Zhang M, Dai J, Pedpradab P, Wang W, Wu J. Cytochalasins from mangrove endophytic fungi Phomopsis spp. xy21 and xy22. Phytochem Lett 2016; 17: 162-166
• 16 Ondeyka J, Hensens OD, Zink D, Ball R, Lingham RB, Bills G, Dombrowski A, Goetz M. L-696, 474, a novel cytochalasin as an inhibitor of HIV-1 protease. II. Isolation and structure. J Antibiot (Tokyo) 1992; 45: 679-685
• 17 Montenegro TG, Rodrigues FA, Jimenez PC, Angelim AL, Melo VM, Rodrigues Filho E, de Oliveira Mda C, Costa-Lotufo LV. Cytotoxic activity of fungal strains isolated from the ascidian Eudistoma vannamei . Chem Biodivers 2012; 9: 2203-2209
• 18 Lingham RB, Hsu A, Silverman KC, Bills GF, Dombrowski A, Goldman ME, Darke PL, Huang L, Koch G, Ondeyka JG, Goetz MA. L-696, 474, a novel cytochalasin as an inhibitor of HIV-1 protease. III. Biological activity. The J Antibiot (Tokyo) 1992; 45: 686-691
• 19 Haidle AM, Myers AG. An enantioselective, modular, and general route to the cytochalasins: synthesis of L-696, 474 and cytochalasin B. Proc Natl Acad Sci U S A 2004; 101: 12048-12053
• 20 Chen LQ, Shen XF, Hu BY, Lin Y, Igbe I, Zhang CG, Zhang GL, Yuan XH, Wang F. Nitric oxide production inhibition and mechanism of phenanthrene analogs in lipopolysaccharide-stimulated RAW 264.7 macrophages. Bioorg Med Chem Lett 2016; 26: 2521-2525
• 21 Gabay C. Interleukin-6 and chronic inflammation. Arthritis Res Ther 2006; 8 (Suppl. 02) S3
• 22 Kim EK, Choi EJ. Compromised MAPK signaling in human diseases: an update. Arch Toxicol 2015; 89: 867-882
• 23 Tak PP, Firestein GS. NF-kappaB: a key role in inflammatory diseases. J Clin Invest 2001; 107: 7-11
• 24 Hu B, Zhang H, Meng X, Wang F, Wang P. Aloe-emodin from rhubarb (Rheum rhabarbarum) inhibits lipopolysaccharide-induced inflammatory responses in RAW 264.7 macrophages. J Ethno 2014; 153: 846-853
• 25 Jouda JB, Mfotie Njoya E, Mbazoa CD, Zhou Z, Meli Lannang A, Wandji J, Shiono Y, Wang F. Lambertellin from Pycnoporus sanguineus MUCL 51321 and its anti-inflammatory effect via modulation of MAPK and NF-kappaB signaling pathways. Bioorg Chem 2018; 80: 216-222
• 26 Qiang Z, Zhou ZY, Peng T, Jiang PZ, Shi N, Njoya EM, Azimova B, Liu WL, Chen WH, Zhang GL, Wang F. Inhibition of TPL2 by interferon-alpha suppresses bladder cancer through activation of PDE4D. J Exp Clin Cancer Res 2018; 37: 288
• 27 Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 2003; 3: 768-780
• 28 Kundu JK, Shin YK, Kim SH, Surh YJ. Resveratrol inhibits phorbol ester-induced expression of COX-2 and activation of NF-kappaB in mouse skin by blocking IkappaB kinase activity. Carcinogenesis 2006; 27: 1465-1474
• 29 Sizemore N, Lerner N, Dombrowski N, Sakurai H, Stark GR. Distinct roles of the Ikappa B kinase alpha and beta subunits in liberating nuclear factor kappa B (NF-kappa B) from Ikappa B and in phosphorylating the p65 subunit of NF-kappa B. J Biol Chem 2002; 277: 3863-3869
• 30 Jouda JB, Kusari S, Lamshoft M, Mouafo Talontsi F, Douala Meli C, Wandji J, Spiteller M. Penialidins A–C with strong antibacterial activities from Penicillium sp., an endophytic fungus harboring leaves of Garcinia nobilis . Fitoterapia 2014; 98: 209-214
• 31 Chemical Computing Group Inc.. MOE. Available at (Accessed 2018): http://www.chemcomp.com
• 32 Halgren T. MMFF94s option for energy minimization studies. J Comp Chem 1999; 20: 720-729
• 33 Becke A. Density-functional exchange-energy approximation with correct asymptotic behavior. Phys Rev 1988; 38: 3098-3100
• 34 Karton A, Tarnopolsky A, Lamere JF, Schatz GC, Martin JM. Highly accurate first-principles benchmark data sets for the parametrization and validation of density functional and other approximate methods. Derivation of a robust, generally applicable, double-hybrid functional for thermochemistry and thermochemical kinetics. J Phys Chem A 2008; 112: 12868-12886
• 35 Perdew J. Density-functional approximation for the correlation energy of the inhomogeneous electron gas. Phys Rev 1986; 33: 8822-8824
• 36 Neese F, Becker U, Ganyushin D, Hansen A, Liakos D, Kollmar C, Koßmann S, Petrenko T, Reimann C, Riplinger C, Sivalingam K, Wezisla B, Wennmohs F. ORCA – an ab initio, density functional and semiempirical program package, version 2.9.1 Max Planck Institute for Bioinorganic Chemistry: Germany. Available at (Accessed 2012): https://orcaforum.cec.mpg.de/
• 37 Yanai T, Tew DP, Handy NC. A new hybrid exchange-correlation functional using the Coulomb-attenuating method (CAM-B3LYP). Chem Phys Lett 2004; 393: 51-57
• 38 Sinnecker S, Rajendran A, Klamt A, Diedenhofen M, Neese F. Calculation of solvent shifts on slectronic g-tensors with the conductor-like screening model (COSMO) and its self-consistent generalization to real solvents (COSMO-RS). J Phys Chem A 2006; 110: 2235-2245
• 39 Barone V, Cossi M. Quantum calculation of molecular energies and energy gradients in solution by a conductor solvent sodel. J Phys Chem A 1998; 102: 1995-2001
• 40 Bruhn T, Schaumloffel A, Hemberger Y, Bringmann G. SpecDis: quantifying the comparison of calculated and experimental electronic circular dichroism spectra. Chirality 2013; 25: 243-249