https://pubs.rsc.org/en/content/articlepdf/2019/NP/C8NP00092A

Marine natural products.

Rate constants for autoxidation propagation of several unsaturated lipids in benzene solution at 37 degrees C and in phosphatidylcholine liposomes were determined by a linoleate radical clock. This radical clock is based on competition between hydrogen atom abstraction by an intermediate peroxyl radical derived from linoleic acid that leads to a trans,cis-conjugated hydroxyoctadecadienoic product and beta-fragmentation of the same peroxyl that gives the trans,trans-product hydroxyoctadecadienoic acid. Rate constants determined by this approach in solution relative to linoleic acid (k(p) = 62 M(-1) s(-1)) were: arachidonic acid (k(p) = 197 +/- 13 M(-1) s(-1)), eicosapentaenoic acid (k(p) = 249 +/- 16 M(-1) s(-1)), docosahexaenoic acid (k(p) = 334 +/- 37 M(-1) s(-1)), cholesterol (k(p)= 11 +/- 2 M(-1) s(-1)), and 7-dehydrocholesterol (k(p)= 2260 +/- 40 M(-1) s(-1)). Free radical oxidations of multilamellar and unilamellar liposomes of various mixtures of glycerophosphatidylcholine molecular species were also carried out. In some experiments, cholesterol or 7-dehydrocholesterol was incorporated into the lipid mixture undergoing oxidation. A phosphatidylcholine bearing a linoleate ester at sn-2 was a component of each liposome peroxidation reaction and the ratio of trans,cis/trans,trans (t,c/t,t)-conjugated diene oxidation products formed from this phospholipid was determined for each oxidation reaction. This t,c/t,t-product ratio from linoleate was used to "clock" liposome constituents as hydrogen atom donors in the lipid bilayer. Application of this lipid bilayer radical clock gives relative autoxidation propagation rate constants of arachidonate (20:4), eicosapentaenoate (20:5), docosahexaenoate (22:6), and 7-dehydrocholesterol to be 115 +/- 7, 145 +/- 8, 172 +/- 13, and 832 +/- 86, respectively, a reactivity trend that parallels the one in solution. We also conclude from the liposome oxidations that linoleate peroxyl radicals at different positions on the eighteen-carbon chain (at C-9 and C-13) have different kinetic properties. This is in contrast to the results of solution oxidations of linoleate in which the C-9 and C-13 peroxyl radicals have similar reactivities. We suggest that peroxyl radical beta-scission depends on solvent polarity and the polarity of the local environment of peroxyl radicals in liposomal oxidations depends on the position of the peroxyl radical on the 18-carbon chain.

/pdf/rate-constants-for-peroxidation-of-polyunsaturated-fatty-582nzah05p.pdf

Rate Constants for Peroxidation of Polyunsaturated Fatty Acids and Sterols in Solution and in Liposomes

Free radical chain oxidation of highly oxidizable 7-dehydrocholesterol (7-DHC), initiated by 2,2'-azobis(4-methoxy-2,4-dimethylvaleronitrile), was carried out at 37 degrees C in benzene for 24 h. Fifteen oxysterols derived from 7-DHC were isolated and characterized with 1D and 2D NMR spectroscopy and mass spectrometry. A mechanism that involves abstraction of hydrogen atoms at C-9 and/or C-14 is proposed to account for the formation of all of the oxysterols and the reaction progress profile. In either the H-9 or H-14 mechanism, a pentadienyl radical intermediate is formed after abstraction of H-9 or H-14 by a peroxyl radical. This step is followed by the well-precedented transformations observed in peroxidation reactions of polyunsaturated fatty acids such as oxygen addition, peroxyl radical 5-exo cyclization, and S(H)i carbon radical attack on the peroxide bond. The mechanism for peroxidation of 7-DHC also accounts for the formation of numerous oxysterol natural products isolated from fungal species, marine sponges, and cactaceous species. In a cell viability test, the oxysterol mixture from 7-DHC peroxidation was found to be cytotoxic to Neuro2a neuroblastoma cells in the micromolar concentration range. We propose that the high reactivity of 7-DHC and the oxysterols generated from its peroxidation may play important roles in the pathogenesis of Smith-Lemli-Opitz syndrome, X-linked dominant chondrodysplasia punctata, and cerebrotendinous xanthomatosis, all of these being metabolic disorders characterized by an elevated level of 7-DHC.

Oxysterols from free radical chain oxidation of 7-dehydrocholesterol: product and mechanistic studies.

This review summarizes new findings concerning the sources and characteristics of various natural products that can be extracted from mangrove-associated microbes over the past three years (January 2011–December 2013). The natural products are discussed with a focus on bioactivity, highlighting the unique chemical diversity of these metabolic products.

Bioactive natural products derived from mangrove-associated microbes

Two new cuparene-type sesquiterpenes, enokipodins C (1) and D (2), were isolated from culture medium of an edible mushroom, Flammulina velutipes, along with enokipodins A (3) and B (4). The structures of 1 and 2 were determined using spectroscopic methods (HRMS, (1)H and (13)C, and 2D NMR). The absolute configuration of enokipodin C was determined from the observed (1)H NMR chemical shifts and NOEs in NOESY experiments after conversion into the corresponding esters with the chiral reagent 2-(2'-methoxy-1'-naphthyl)-3,4-dichlorobenzoic acid. All the metabolites showed antimicrobial activity against a fungus, Cladosporium herbarum, and Gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus.

http://www.caspmi.cn/zhanglab/papers/2011/Enokipodins.pdf

Antimicrobial cuparene-type sesquiterpenes, enokipodins C and D, from a mycelial culture of Flammulina velutipes.

Eight new sterols, 5α, 8α-epidioxy-(22E, 24R)-23-methylergosta-6, 22-dien-3β-ol (1), 3β, 5α, 9α-trihydroxy-(22E, 24R)-23-methylergosta-7, 22-dien-6-one (2), 3β, 5α, 9α-trihydroxy-(24S)-ergost-7-en-6-one (3), 3β, 5α, 9α, 14α-tetrahydroxy-(22E, 24R)-ergosta-7, 22-dien-6-one (4), (22E, 24R)-ergosta-7, 22-diene-3β, 5α, 6α, 9α-tetrol (5), 5α, 9α-epidioxy-3β-hydroxy-(22E, 24R)-ergosta-7, 22-dien-6-one (6), 5α, 9α-epidioxy-3β-hydroxy-(24S)-ergost-7-en-6-one (7) and 5α, 6α-epoxy-(22E, 24R)-ergosta-8, 22-diene-3β, 7β, 14α-triol (8), have been isolated from five edible mushrooms, Lentinus edodes, Flammulina velutipes, Hypsizigus marmoreus, Pleurotus ostreatus and Pholiota nameko together with fifteen known ones (9-23), of which two (16 and 17) are reported for the first time from a fungal source. The structures of these new compounds were elucidated on the basis of their spectral data.

Akihiro Masaki

Papers

Sterol Constituents from Five Edible Mushrooms