Studies On Secondary Metabolites From The Co-cultivation Of Two Fungi And Their Anti-Parkinson’s Disease Activity

This study mainly consists of three parts:the first part is drug deficiency in Parkinson’s disease and the promotion of new secondary metabolites generation under fungi coculture;The second part is a study on the chemical composition under coculture of Alternaria brassicicola and Penicillium granulosum;The third part is to carry out biological activity research in anti-Parkinson’s disease on new compounds obtained from coculture.In this study,Alternaria brassicicola and Penicillium granulosum were selected,which were solid fermented by lab environment for 28 days and extracted with anhydrous methanol.In turn,the extracts were fractionated and purified using modern chromatographic techniques,combined with 1D and 2D NMR,HRESIMS,IR,UV,calculated ECD,calculated ¹³C NMR spectrum,chemical derivatization,and X-ray methods,and aligned with the literature,leading to the final determination of twenty-three compounds,nine of which were novel compounds,including four steroids（2-5）,three fusicoccane diterpenoids（7-9）,and compound 7 was a novel tetracyclic fusicoccane diterpene,twoα-Pyranone derivatives（20,23）.Other known compound structure types also include Conidiogenones type diterpenes.New compounds were investigated for their pharmacological activity and mechanism against Parkinson’s disease.Initially,fusicoccane diterpenoids 7,8 were found to be well protective against H₂O₂induced oxidative damage in SH-SY5Y cells.Steroidal compound 4in MPP⁺-induced PD model of SH-SY5Y could finally exert protective effects on SH-SY5Y neuronal cells by blocking the assembly of NLRP3 inflammasome and then inhibiting pyroptosis.Theα-pyrone derivative compound 20,in the PD model of SH-SY5Y cells induced by 6-OHDA,could exert protective effects on SH-SY5Y neuronal cells by inhibiting the ferroptosis pathway.In summary,we identified twenty-three compounds isolated from them by establishing a fungal coculture model to stimulate the production of novel secondary metabolites,containing nine novel compounds,including four steroids,three fusicoccane type diterpenes,and twoα-pyrone derivatives.And they were screened for anti-PD activity and found the neuroprotective activity exhibited by compounds 4,7,8,20 in different models,which provided a novel theoretical rationale and experimental basis in leading drug discovery of anti-PD.