Berkeley Pit Lake is a 1500 ft. deep abandoned open-pit copper mine filled with over 1140 billion liters of acidic, metal-sulfate contaminated water. This harsh environment is proving to be a source of unusual microorganisms that produce novel bioactive compounds. We recently reported the structures of berkeleydione (1) and berkeleytrione, two novel hybrid polyketide-terpenoid metabolites isolated from a deep water Penicillium sp. growing in Berkeley Pit lake. In this paper we report the structures of three new compounds, berkeleyacetals A-C (3-5) isolated from extracts of this fungus. The structures of these compounds were deduced by comparison of mass spectral and NMR data to that of berkeleydione 1.The Berkeley Pit Lake System is part of the largest EPA Superfund site in North America. Berkeley Pit Lake contains roughly 1140 billion liters of acidic (pH 2.5), metal-sulfate rich water that sits poised at the head of the Columbia River ecosystem. 1 We are studying the secondary metabolism of the culturable microbes isolated from Pit water and sediment samples. Several interesting compounds have been isolated from these microbes based on their activities in different bioassay guided fractionation schemes. [2][3][4][5][6] One of the first microbes to be studied from this environment was isolated from a depth of 885 feet and was subsequently identified as Penicillium rubrum Stoll based on rRNA sequencing alignment data (300 base pairs). 3 The fungus was grown in acidified potato dextrose broth (pH 2.7, still) for 21 days. At harvest time the fungus was killed by the addition of MeOH, the mycelia removed by gravity filtration and the filtrate extracted with CHCl 3 . The CHCl 3 extract inhibited both MMP-3 and caspase-1 in assay systems so it was fractionated by LH-20 column chromatography followed by HPLC to yield berkeleydione (1) and berkeleytrione (2). 3 The 1 H NMR spectra from all of the column fractions were examined for evidence of related analogs. Promising candidates were purified and tested for activity. The three new berkeleyacetals 3-5 were isolated based on this "NMR guided" fractionation as well as their activity in both enzyme bioassays. We report here the isolation and characterization of berkeleyacetals A-C, (3-5). The structures were established based upon extensive spectroscopic data and a single crystal X-ray structure for 1.We are exploring the possibility of isolating compounds with anticancer potential using in vitro signal transduction bioassay systems. Two enzyme inhibition schemes have shown promising correlation to the National Cancer Institutes Developmental Therapeutics Program human cancer cell line assay. To date, compounds isolated in our lab based on these assays have shown activity against non-small cell lung cancer 3 and ovarian cancer 4 . Matrix metalloproteinases (MMP) hydrolyze the extracellular matrix of connective tissues and basement membranes. 7, 8 MMP-3 is up-regulated in many tumors 9, 10 and can induce *To whom correspondence should be addressed. Tel: (406) Casp...

We previously reported several novel bioactive hybrid polyketide-terpenoid metabolites from a deep water Penicillium rubrum isolated from Berkeley Pit Lake, Butte, Montana. In this paper we report the structures of four new amides, berkeleyamides A-D (1, 4, 5, 7), isolated from extracts of this fungus. The structures of these compounds were deduced by analysis of NMR data, chemical derivatization, and comparison of their spectroscopic data to those of known compounds.

The Berkeley Pit, an acid mine waste lake, is a source of extremophilic microorganisms that produce interesting bioactive compounds. We have previously reported the isolation of berkeleydione 1, berkeleytrione 2, the berkeleyacetals and the berkeleyamides from the Pit Lake fungus Penicillium rubrum. In this paper we report the isolation and characterization of berkeleyones A-C (4, 5 and 7) as well as previously described preaustinoid A (3) and A1(6) from this same fungus. These compounds were evaluated as inhibitors of the signaling enzyme caspase-1 and as potential inhibitors of interleukin 1-β production by inflammasomes in induced THP-1 cell line assays.