Penicipyrrolidines A−N, pyrrolidine derivatives with inhibitory effects on EMT and fibroblast activation from the mangrove-derived fungus Penicillium sp. DM27

Abstract

An investigation of the mangrove-derived fungus Penicillium sp. DM27 led to the isolation of 19 new compounds, including three pairs of piperidinone enantiomers ( ±)-1, ( ±)-2, and ( ±)-3, two pairs of pyrrolidinone enantiomers ( ±)-4 and ( ±)-5, and nine pyrrolidine derivatives 6−14. The structures of 1−14 were elucidated through NMR and HRESIMS analysis, coupled with experimental and calculated ECD spectroscopy and the modified Mosher method. Quantitative real time PCR and Western bolt analyses revealed that 11 blocked EMT in TGF-β1-treated HK-2 cells and suppressed fibroblast activation in TGF-β1-stimulated NIH-3T3 cells. Molecular simulations demonstrated that compound 11 could dock ADAM17, showing a high negative binding affinity. Additionally, the overexpression of ADAM17 by lentiviral infection triggered renal tubular EMT, while compound 11 suppressed this process. Overall, our research suggests that pyrrolidine derivatives may be potential therapeutic agents for the treatment of fibrotic kidney disease.

Keywords

Penicillium sp. / Pyrrolidine / Pyrrolizidine alkaloids / Renal fibrosis / Fibroblast activation

References

Bai Y, Wang W, Yin P, Gao J, Na L, Sun Y, Wang Z, Zhang Z, Zhao C. Ruxolitinib alleviates renal interstitial fibrosis in UUO Mice. Int Biol Sci, 2020, 16: 194-203

Cantin A, Moya P, Castillo MA, Primo J, Miranda MA, Primo-Yufera E. Isolation and synthesis of N-(2-methyl-3-oxodec-8-enoyl)-2-pyrroline and 2-(hept-5-enyl)-3-methyl-4-oxo-6,7,8,8a-tetrahydro-4H- pyrrolo[2,1-b]-1,3-oxazine. Two new fungal metabolites with in-vivo anti-juvenile-hormone and insecticidal activity. Eur J Org Chem, 1999, 1: 221-226

Chen XP, Ma LF, Zhan ZJ. A new pyrrolizidine alkaloid from Penicillium expansum. J Chem Res, 2017, 41: 93-94

Chen DQ, Feng YL, Cao G, Zhao YY. Natural products as a source for antifibrosis therapy. Trends Pharmacol Sci, 2018, 39: 937-952

Chen S, Cai R, Liu Z, Cui H, She Z. Secondary metabolites from mangrove-associated fungi: Source, chemistry and bioactivities. Nat Prod Rep, 2022, 39: 560-595

Chen XY, Shi M, Yang LT, Guo F, Liang Y, Ma L, Fu P. Phenylethanoid glycoside verbascoside ameliorates podocyte injury of diabetic kidney disease by regulating NR4A1-LKB1-AMPK signaling. Acta Mater Med, 2023, 2: 72-83.

Cimmino A, Masi M, Evidente M, Superchi S, Evidente A. Application of Mosher's method for absolute configuration assignment to bioactive plants and fungi metabolites. J Pharm Biomed, 2017, 144: 59-89

Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Scalmani G, Barone V, Mennucci B, Petersson GA, Nakatsuji H, Caricato M, Li X, Hratchian HP, Izmaylov AF, Bloino J, Zheng G, Sonnenberg JL, Hada M, Ehara M, et al. Gaussian 09, Revision A.02, 2009. Wallingford: Gaussian Inc.

Hao HP, Zheng X, Wang GJ. Insights into drug discovery from natural medicines using reverse pharmacokinetics. Trends Pharmacol Sci, 2014, 35: 168-177

Huang H, Feng X, Xiao Z, Liu L, Li H, Ma L, Lu Y, Ju J, She Z, Lin Y. Azaphilones and p-terphenyls from the mangrove endophytic fungus Penicillium chermesinum (ZH4-E2) isolated from the South China Sea. J Nat Prod, 2011, 74: 997-1002

Jager KJ, Kovesdy C, Langham R, Rosenberg M, Jha V, Zoccali C. A single number for advocacy and communication-worldwide more than 850 million individuals have kidney diseases. Kidney Int, 2019, 96: 1048-1050

Kalinowski HO, Berger S, Braun S. Carbon-13 NMR spectroscopy, 1988. Chichester: Wiley 293.

Leshchenko EV, Antonov AS, Dyshlovoy SA, Berdyshev DV, Hauschild J, Zhuravleva OI, Borkunov GV, Menshov AS, Kirichuk NN, Popov RS, Gerasimenko AV, Udovenko AA, Graefen M, Bokemeyer C, Von Amsberg G, Yurchenko AN. Meroantarctines A−C, meroterpenoids with rearranged skeletons from the alga-derived fungus Penicillium antarcticum KMM 4685 with potent p-glycoprotein inhibitory activity. J Nat Prod, 2022, 85: 2746-2752

Li K, Chen S, Pang X, Cai J, Zhang X, Liu Y, Zhu Y, Zhou X. Natural products from mangrove sediments-derived microbes: structural diversity, bioactivities, biosynthesis, and total synthesis. Eur J Med Chem, 2022, 230: 114-117

Liu Y. New insights into epithelial-mesenchymal transition in kidney fibrosis. J Am Soc Nephrol, 2010, 21: 212-222

Liu Y. Cellular and molecular mechanisms of renal fibrosis. Nat Rev Nephrol, 2011, 7: 684-696

Moya P, Cantin A, Castillo MA, Primo J, Miranda MA, Primo-Yufera E. Isolation, structural assignment, and synthesis of N-(2-methyl-3-oxodecanoyl)-2-pyrroline, a new natural product from Penicillium brevicompactum with in Vivo anti-juvenile hormone activity. J Org Chem, 1998, 63: 8530-8535

Moya P, Cantin A, Miranda MA, Primo J, Primo-Yufera E. Synthesis and biological evaluation of new analogues of the active fungal metabolites N-(2-methyl-3-oxodecanoyl)-2-pyrroline and N-(2-methyl-3-oxodec -8-enoyl)-2-pyrroline. J Agr Food Chem, 1999, 47: 3866-3871

Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. J Nat Prod, 2020, 83: 770-803

Ohtani I, Kusumi T, Kashman Y, Kakisawa H. High-field FT NMR application of Mosher's method. The absolute configurations of marine terpenoids. J Am Chem Soc, 1991, 113: 4092-4096

Peng SG, Liu XN, Sura MB, Yan YM, Cheng YX. Mantidisflavin A: A riboflavin derivative featuring a 6/6/6/5/5 skeleton from the egg cases of the insect Tenodera sinensis Saussure and its anti-renal fibrosis activity. Org Lett, 2024, 26: 1316-1320

Romagnani P, Remuzzi G, Glassock R, Levin A, Jager KJ, Tonelli M, Massy Z, Wanner C, Anders H. Chronic kidney disease. J Nat Rev Dis Primers, 2017, 3: 17088

Ruiz-Ortega M, Rayego-Mateos S, Lamas S, Ortiz A, Rodrigues-Diez RR. Targeting the progression of chronic kidney disease. Nat Rev Nephrol, 2020, 16: 269-288

Sadorn K, Saepua S, Boonyuen N, Laksanacharoen P, Rachtawee P, Pittayakhajonwut P. Antimicrobial activity and cytotoxicity of polyketides isolated from the mushroom Xerula sp. BCC56836. RSC Adv, 2016, 6: 94510-94523

Sasaki M, Tsuda M, Sekiguchi M, Mikami Y, Kobayashi J. Perinadine A, a novel tetracyclic alkaloid from marine-derived fungus Penicillium citrinum. Org Lett, 2005, 7: 4261-4264

Shao CL, Wang CY, Gu YC, Wei MY, Pan JH, Deng DS, She ZG, Lin YC. Penicinoline, a new pyrrolyl 4-quinolinone alkaloid with an unprecedented ring system from an endophytic fungus Penicillium sp. Bioorg Med Chem Lett, 2010, 20: 3284-3286

Sheng L, Zhuang S, Zhuang S. New insights into the role and mechanism of partial epithelial-mesenchymal transition in kidney fibrosis. Front Physiol, 2020, 11: 569322.

Singh R, Kaundal RK, Zhao B, Bouchareb R, Lebeche D. Resistin induces cardiac fibroblast-myofibroblast differentiation through JAK/STAT3 and JNK/c-Jun signaling. Pharmacol Res, 2021, 167: 105414.

Tsuda M, Sasaki M, Mugishima T, Komatsu K, Sone T, Tanaka M, Mikami Y, Kobayashi J. Scalusamides A−C, new pyrrolidine alkaloids from the marine-derived fungus Penicillium citrinum. J Nat Prod, 2005, 68: 273-276

Wang HC, Ke TY, Ko YC, Lin JJ, Chang JS, Cheng YB. Anti-inflammatory azaphilones from the edible alga-derived fungus Penicillium sclerotiorum. Mar Drugs, 2021, 19: 529

Weiskirchen R, Weiskirchen S, Tacke F. Organ and tissue fibrosis: Molecular signals, cellular mechanisms and translational implications. Mol Aspects Med, 2019, 65: 2-15

Wu J, Zhang H, He LM, Xue YQ, Jia J, Wang SB, Zhu KK, Hong K, Cai YS. A new fusicoccane-type norditerpene and a new indone from the marine-derived fungus Aspergillus aculeatinus WHUF0198. Chem Biodivers, 2021, 18: e2100562.

Wu J, Wang F, He LM, Zhou SY, Wang F, Wang SB, Jia J, Hong K, Cai YS. Aculeaquamide A, cytotoxic paraherquamide from the marine fungus Aspergillus aculeatinus WHUF0198. Nat Prod Res, 2022, 36: 4388-4393

Yang JK, Zhang B, Gao T, Yang MY, Zhao GZ, Zhu HJ, Liu L, Cao F. A pair of enantiomeric 5-oxabicyclic[4.3.0]lactam derivatives and one new polyketide from the marine-derived fungus Penicillium griseofulvum. Nat Prod Res, 2018, 32: 2366-2369

Zeisberg M, Neilson EG. Mechanisms of tubulointerstitial fibrosis. J Am Soc Nephrol, 2010, 21: 1819-1834

Zhang YH, Li L, Li YQ, Luo JH, Li W, Li LF, Zheng CJ, Cao F. Oxalierpenes A and B, unusual indole-diterpenoid derivatives with antiviral activity from a marine-derived strain of the fungus Penicillium oxalicum. J Nat Prod, 2022, 85: 1880-1885

Zhou ZF, Kurtan T, Yang XH, Mandi A, Geng MY, Ye BP, Taglialatela-Scafati O, Guo YW. Penibruguieramine A, a novel pyrrolizidine alkaloid from the endophytic fungus Penicillium sp. GD6 associated with Chinese mangrove Bruguiera gymnorrhiza. Org Lett, 2014, 16: 1390-1393

Zhou R, Liao J, Cai D, Tian Q, Huang E, Lue T, Chen SY, Xie WB. Nupr1 mediates renal fibrosis via activating fibroblast and promoting epithelial-mesenchymal transition. FASEB J, 2021, 35: e21381.