Protease inhibitors from three fishpond water blooms of Microcystis spp.

Four new natural products, micropeptin GH979 (1), microginin GH787 (2), micropeptin HM978 (3), and micropeptin HA983 (4), as well as 10 known protease inhibitors and hepatotoxins, were isolated from the hydrophilic extract of three samples of cyanobacteria (Microcystis spp.) that were collected from fishponds in Kibbutz Giva’at Haim, Kibbutz Hama’apil, and Kibbutz Gan Shmuel. The structures of the pure natural products were elucidated using spectroscopic methods, including UV, 1D and 2D NMR, and MS techniques. The absolute configuration of the chiral centers of the compounds was determined using Marfey’s method. The inhibitory activity of the compounds was determined for the serine proteases: trypsin, chymotrypsin, thrombin and elastase, and the metalloprotease, aminopeptidase N.     

Three novel metabolites from a bloom of the cyanobacterium Microcystis sp.

Three new metabolites, microphycin AL828, microguanidine AL772, and microginin AL584 and three known metabolites, anabaenopeptin F, oscillamide Y, and microcin SF608 were isolated from the extracts of a Microcystis sp. bloom collected in Alonim reservoir, Valley of Armagedon, Israel. The planar structure of the compounds was determined by homonuclear and inverse-heteronuclear 2D-NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers of the amino acids was studied using Marfey's method for HPLC.     

Two new microcyclamides from a water bloom of the cyanobacterium Microcystis sp.

Two new thiazole-containing cyclic hexapeptides, microcyclamides MZ602 and MZ568, were isolated from the hydrophilic extract of the cyanobacterium Microcystis sp. The structures of the pure natural products were elucidated using spectroscopic methods including UV, 1D and 2D NMR, and MS techniques. The absolute configuration of the chiral centres of the modified cyclic peptides was determined using Marfey’s method for HPLC. The microcyclamides have been evaluated for their cytotoxicity against leukemia cell lines and inhibition of serine proteases.     

Eight micropeptins from a Microcystis spp. bloom collected from a fishpond near Kibbutz Lehavot HaBashan,Israel

Eight new micropeptin-type cyclic depsipeptides, micropeptins LH920, LH1021, LH1048, LH1062, LH911A, LH911B, LH911C, and LH925, along with five known micropeptins, three known anabaenopeptins and two known microcystins (LR and RR), were isolated from a water bloom biomass of Microcystis spp. assembly that was collected from a fishpond near Kibbutz Lehavot HaBashan, Israel. The structures of the pure compounds were elucidated using 1D and 2D NMR techniques, as well as high-resolution mass spectrometry. The stereochemistry of the new natural products was determined using Marfey's method for amino acids. The inhibitory activity of the compounds was determined for the serine proteases, trypsin, and chymotrypsin.     

Micropeptins from Microcystis sp. collected in Kabul Reservoir,Israel

Three new micropeptins, micropeptin KR1030, KR1002 and KR998 and the known microcyclamide GL546A were isolated from the extract of Microcystis sp. bloom material collected in Kabul Reservoir, Israel. The planar structures of the compounds were determined by homonuclear and inverse-heteronuclear 2D-NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centres of the amino acids was studied using Marfey's method for HPLC. The inhibitory activity of the compounds was determined for the serine proteases: trypsin, chymotrypsin and elastase.     

Novel thiazole and oxazole containing cyclic hexapeptides from a waterbloom of the cyanobacterium Microcystis sp.

Eight novel thiazole and oxazole containing cyclic peptides, microcyclamides GL616, GL582, GL614A, GL614B, GL614C, GL546A, GL546B, and GL628, as well as the known microcyclamide A, were isolated from the hydrophylic extract of a Microcystis sp. water-bloom collected in Gilboa reservoir, Valley of Armageddon, Israel. The planar structure of the compounds was determined by homonuclear and inverse-heteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers of the amino acids was studied using Marfey's method for HPLC following ozonolysis, hydrolysis and derivatization with Marfey's reagent. This is the first example where acidic and modified amino acids are incorporated in this group of ribosomally biosynthesized metabolites.     

Banyasin A and banyasides A and B, three novel modified peptides from a water bloom of the cyanobacterium Nostoc sp.

Three new modified peptides banyasin A, banyaside A and banyaside B were isolated from the hydrophilic extract of a natural bloom of the cyanobacterium Nostoc sp. The planar structure of the new compounds was determined by homonuclear and inverse-heteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. Banyasides A and B, are structurally closely related to the cyanobacteria metabolite, suomilide and to the sponge derived, dysinosins. The absolute configuration of the asymmetric centers was studied using Marfey's method for HPLC. Banyaside A and B were found to be trypsin and thrombin inhibitors.     

New microviridins from a water bloom of the cyanobacterium Microcystis aeruginosa

Three new microviridins namely, SD1684 (1), SD1634 (2), and SD1652 (3), were isolated from the hydrophilic extract of Microcystis aeruginosa. The planar structures of compounds 1-3 were determined by homonuclear and inverse-heteronuclear 2D-NMR techniques as well as by high-resolution mass spectrometry. The absolute configuration of the asymmetric centers was studied using Marfey's method for HPLC. Compounds 1-3 contain l-threo-β-hydroxy aspartic acid as a building block of the peptide chain. This is the first example where microviridins contain non-proteinogenic amino acid in their structure. Compound 2 is a mild serine protease inhibitor.     

Micropeptins from Microcystis aeruginosa collected in Dalton reservoir, Israel

Three new micropeptins, micropeptin DR1056, DR1060 and DR1006 and three known metabolites, micropeptin SF909, aeruginosins 298A and B were isolated from the extracts of a Microcystis aeruginosa bloom material collected in Dalton reservoir, Israel. The planar structure of the compounds was determined by homonuclear and inverse-heteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers of the amino acids was studied using Marfey’s method for HPLC. The inhibitory activity of the compounds was determined for the serine proteases: trypsin, chymotrypsin, and elastase and for amino peptidase N.     

Three novel anabaenopeptins from the cyanobacterium Anabaena sp.

Three new cyclic peptides, anabaenopeptins NZ825, NZ841, and NZ857, were isolated from the hydrophilic extract of the cultured cyanobacterium Anabaena sp. The planar structure of the compounds was determined by homonuclear and inverse-heteronuclear 2D-NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers was studied using Marfey's method for HPLC. This is the first report of anabaenopeptins that contain N-methyl glycine instead of the common N-methyl alanine. The incorporation of N-methyl glycine into the cyclic portion of the compounds results in their appearance as a mixture of two, equally stable, conformers, instead of the one distinct conformer in anabaenopeptins that contain N-methyl alanine or N-methyl homotyrosine. The three compounds were tested for inhibition of serine proteases and found to be not active.     

Three aeruginosins and a microviridin from a bloom assembly of Microcystis spp. collected from a fishpond near Kibbutz Lehavot HaBashan,Israel

Four new metabolites, aeruginosins LH650A (1), LH650B (2), and LH606 (3), and microviridin LH1667 (4) were isolated along with the two known aeruginosins 98-A and DA495B, and previously isolated metabolites, from a bloom material of Microcystis spp., collected from a fishpond near Kibbutz Lehavot HaBashan, Israel. Their structures were elucidated by a combination of various spectroscopic techniques; primarily NMR and MS, while the absolute configuration of the chiral centers was determined by Marfey's method and chiral-phase HPLC. The structures and biological activities of the four new metabolites are described.     

Marine bacterial inhibitors from the sponge-derived fungus Aspergillus sp.

Chromatographic separation of a crude extract obtained from the fungus Aspergillus sp., isolated from the Mediterranean sponge Tethya aurantium, yielded a new tryptophan derived alkaloid, 3-((1-hydroxy-3-(2-methylbut-3-en-2-yl)-2-oxoindolin-3-yl)methyl)-1-methyl-3,4-dihydrobenzo[e][1,4]diazepine-2,5-dione (1), and a new meroterpenoid, austalide R (2), together with three known compounds (3–5). The structures of the new compounds were unambiguously elucidated on the basis of extensive one and two-dimensional NMR (¹H, ¹³C, COSY, HMBC, and ROESY) and mass spectral analysis. Interestingly, the compounds exhibited antibacterial activity when tested against a panel of marine bacteria, with 1 selectively inhibiting Vibrio species and 2 showing a broad spectrum of activity. In contrast, no significant activity was observed against terrestrial bacterial strains and the murine cancer cell line L5178Y.     

Protease inhibitors from a Slovenian Lake Bled toxic waterbloom of the cyanobacterium Planktothrix rubescens

Three new protease inhibitors, planktopeptin BL1125, planktopeptin BL843 and planktopeptin BL1061 were isolated, along with three known compounds, anabaenopeptin A, anabaenopeptin B and anabaenopeptin F from the hydrophilic extract of Planktothrix rubescens. The planar structure of the new compounds was determined by homonuclear and inverse-heteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers was studied using Marfey's method for HPLC and by comparison of the acid hydrolysate with authentic samples on a chiral HPLC column. The new peptides were found to be elastase and chymotrypsin inhibitors.     

Pompanopeptins A and B, new cyclic peptides from the marine cyanobacterium Lyngbya confervoides

A new 3-amino-6-hydroxy-2-piperidone (Ahp) containing peptolide, pompanopeptin A (1), and a novel N-methyl-2-amino-6-(4′-hydroxyphenyl)hexanoic acid (N-Me-Ahpha) containing cyclic pentapeptide connected with a sixth amino acid residue via a rare ureido linkage, pompanopeptin B (2), were isolated from the marine cyanobacterium Lyngbya confervoides collected from the southeastern coast of Florida. Their planar structures were determined by a combination of NMR spectroscopic analysis and mass spectrometry. The absolute configurations were established using advanced Marfey's method and chiral HPLC analysis of the chemical degradation products. Compound 1 selectively inhibited trypsin over elastase and chymotrypsin, with an IC₅₀ value of 2.4 μM; selectivity is conferred by an arginine residue in the cyclic core.     

Revised structure and structure-activity relationship of bisebromoamide and structure of norbisebromoamide from the marine cyanobacterium Lyngbya sp.

Novel potent cytotoxic peptides bisebromoamide (1) and norbisebromoamide (2) have been isolated from the marine cyanobacterium Lyngbya sp. The planar structure of these peptides was elucidated through the extensive application of 1D and 2D NMR techniques. The absolute stereostructure of 1 was determined by chemical degradation followed by chiral HPLC analysis. Recently, Tao and co-workers achieved synthesis of bisebromoamide, and the configuration of thiazoline moiety was revised. We re-investigated the stereochemistry of thiazoline moiety of 1. The structure-activity relationships of bisebromoamide (1) were investigated with the use of natural and synthetic analogs. Furthermore, bisebromoamide (1) potently inhibited protein kinase: the phosphorylation of ERK in NRK cells by PDGF-stimulation was selectively inhibited by treatment with 10-0.1 μM of 1.     

Carbamidocyclophanes F and G with anti-Mycobacterium tuberculosis activity from the cultured freshwater cyanobacterium Nostoc sp.

Two new (1 and 2) and three known (3–5) carbamidocyclophanes were isolated from a cultured freshwater cyanobacterium Nostoc sp. (UIC 10274) obtained from a sample collected at Des Plaines, Illinois. Their planar structures and stereoconfigurations were determined by extensive spectroscopic analysis including 1D/2D NMR experiments, HRESIMS as well as CD spectroscopy. Carbamidocyclophane F (1) showed potent anti-Mycobacterium tuberculosis activity in the microplate Alamar blue assay and low-oxygen-recovery assay with MIC values of 0.8 and 5.4 μM, respectively. Carbamidocyclophane F (1) also displayed antimicrobial activities against the gram positive bacteria Staphylococcus aureus and Enterococcus faecalis with MIC values of 0.1 and 0.2 μM, respectively. Carbamidocyclophane F (1) and Carbamidocyclophane G (2) both showed antiproliferative activity against MDA-MB-435 and HT-29 human cancer cell lines with IC₅₀ values in the range from 0.5 to 0.7 μM.     

Hapalocyclamide: a novel phytotoxic hexapeptide of the cyanobacterium Hapalosiphon sp.

Hapalocyclamide, a novel oxazole-, thiazole- and thiazoline-containing cyclic hexapeptide, was isolated from the terrestrial cyanobacterium Hapalosiphon sp., and which showed phytotoxic activity on lettuce seedling growth. The gross structure of hapalocyclamide was established from spectroscopic data and chemical degradation. The absolute stereochemistry was determined by Marfey’s analysis. Hapalocyclamide was established as cyclo-thiazole-l-alanine-oxazole-d-alanine-d-thiazoline-d-phenylalanine.     

Discovery of novel sesquiterpenoids from a gorgonian Menella sp.

A novel sesquiterpenoid, menelloide A (1), which was found to possess a new carbon skeleton, and a new guaiane-type sesquiterpenoid, menelloide B (2), along with (+)-chloranthalactone B (3), an enantiomer of the known sesquiterpenoid, chloranthalactone B (4), were isolated from a gorgonian coral identified as Menella sp. The structures of sesquiterpenoids 1-3 were established by spectroscopic methods and by comparison of the data with those of related metabolites. Sesquiterpenoids 1 and 3 displayed inhibitory effects on the generation of superoxide anion by human neutrophils.     

Acremines H-N, novel prenylated polyketide metabolites produced by a strain of Acremonium byssoides

Five novel metabolites, acremines H-N, have been isolated from malt extract-peptone-glucose agar cultures of a strain of Acremonium byssoides. Their structures and stereochemistry were elucidated using a combination of ¹³C and ¹H homo and heteronuclear 2D NMR experiments. Acremines H-N inhibited the germination of sporangia of Plasmopara viticola.     

Zooxanthellamide A, a novel polyhydroxy metabolite from a marine dinoflagellate of Symbiodinium sp.

A novel polyhydroxy metabolite named Zooxanthellamide A, C₁₂₈H₂₂₂N₂O₅₄S₂, was isolated from a cultured marine dinoflagellate, Symbiodinium sp., and the chemical structure was determined by spectroscopic methods.