Camosic acid from rosemary

Mace and colleagues have examined the ability of camosol and camosic acid from rosemary as well as the synthetic dithiolethione, oltipraz, to block the formation of DNA adducts, and their effects on the expression of phase I and phase II enzymes. It was found that both rosemary extracts and oltipraz inhibited BaP- or aflatoxin Bi-induced DNA adduct formation by efficiently inhibiting CYP activities and inducing the expression of GST. Treatment of female CD-I mice with a 2% methanol extract of rosemary in AIN-76A diet for 3 weeks increased the liver microsomal 2-hydroxylation of estradiol and estrone by approximately 150%, increased their 6-hydroxylation by approximately 30%, and inhibited the 16a-hydroxylation of estradiol by approximately 50%. The same treatment of rosemary also stimulated the liver microsomal glucuronidation of estradiol and estrone by 54 to 67% and 37 to 56%, respectively. In additional studies, feeding 2% rosemary diet to ovariectomized CD-I mice for 3 weeks inhibited the uterotropic action of estradiol and estrone by 35 to 50% compared with animals fed a control diet. 

Hops resin 150 mg hops aipha acids (from hops [humulus lupulus, ieaves, flowers] extract) 10 iU naturai Vitamin E (from 8 mg d-aipha tocopheroi acetate) 0.5 mg astaxanthin (from Haematococcus pluvialis aigai extract) 1.5 mg camosic acid (from rosemary [Rosemarinus officinalis, ieaves, flowers] extract) 100 mg olive oil... 

Bailey, D, Richheimwr, S, Bank, V and King, B (1999) High purity camosic acid from rosemary and sage extracts by pH-controlled precipitation. US Patent 5,859,293. 

Hopia, A.I., Huang, S.-W., Schwarz, K., German, J.B. and Frankel, E.N. 1996. Effect of different lipid systems on antioxidant activity of rosemary constituents camosol and camosic acid with and without alpha-xocopherol. J. Agric. Food Chem. 44 2030-2036. Houlihan, C.M., Ho, C.-T. and Chang, S.S. 1984 Elucidation of the chemical structure of a novel antioxidant, rosmaridiphenol, isolated from rosemary. J. Am. Oil Chem. Soc. 61 1036-1039. 

Several phenolic diterpenoids with antioxidant activities were isolated from rosemary leaves. These include camosol, camosic acid, rosmanol, isorosmanol, and epirosmanol. Camosol showed potent antioxidative activity as revealed by scavenging a,a-diphenyl-(3-picrylhydrazyl free radicals and protection of oxidative DNA damage. In chronic inflammation, cytokines induce the production of nitric oxide, which is converted to DNA-damaging and carcinogenic peroxynitrite. Peroxynitrite is a cytotoxicant with... 

Masuda, T., Inaba, Y. and Takeda, Y. Antioxidant mechanism of camosic acid Structural identification of two oxidation products. /. Agric. Food Chem. 49, 5560-5565 (2001). Masuda, T., Inaba, Y., Maekawa, T., Takeda, Y., Tamura, H. and Yamaguchi, H. Recovery mechanism of the antioxidant activity from camosic acid quinone, an oxidized sage and rosemary antioxidants. J. Agric. Food Chem. 50, 5863-5869 (2002). 

Common sage and rosemary (see Table 8.32), plants of the Lami-aceae family, contain the diterpenes camosic acid, also known as rosmaricin (8-259), derived from ent-caurene, and bitter carnosol (picrosalvin, 8-260), which are potent antioxidants. Carnosic acid is a major component of fresh rosemary tops (1-2%), but is unstable and is enzymatically transformed into carnosol. These two diterpenoids represent about 15% w/w of plants haulm extracts and exhibit about 90% of extract antioxidant activity. Other transformation products of carnosic acid are rosmanol (7a-hydroxy derivative, 8-261), epirosmanol (7P-isomer, 8-262) and similar compounds. 

Frankel et al. (1996a) also observed that camosol and camosic acid were significantly more active antioxidants in oil-in-water emulsions buffered at pH 4 or 5 than at pH 7. Hopia et al. (1996) reported that camosic acid was a better antioxidant than camosol in methyl linoleate, but that the reverse was tme in linoleic acid. In methyl linoleate or linoleic acid, camosic acid and camosol were more active than a-tocopherol. However, a-tocopherol was more active in bulk corn oil triacylglycerols, followed by camosic acid and camosol. The oxidative stability of rapeseed oil treated with rosemary extracts correlated well with the content of camosic acid, but not with content of camosol (Trojakova et al, 2001). In all emulsified systems, a-tocopherol exhibited more antioxidant activity than camosol or camosic acid (Hopia et al., 1996). In bulk com oil, the addition of camosol to a-tocopherol had a negative impact on the antioxidant activity of a-tocopherol, whereas camosic acid enhanced a-tocopherol activity (Hopia et al, 1996). Both camosic acid and camosol disappeared from the test system more quickly than a-tocopherol (Hopia et al, 1996). 

Camosol and ursolic acid were isolated from rosemary as described (5). Camosic acid and rosmarinic acid were isolated from the ground dried leaves of rosemary, sequentially, by hexane and n-butanol extractions. The final products were purified by column chromatography on silica gel (2). LPS (lipopolysaccharide, Escherichia coli 026 B6), sulfanilamide and naphthyl-ethylenediamine dihydrochloride were purchased from Sigma Chemical Co. (St. Louis, MO). Isotopes were obtained from Amersham (Arlington Heights, IL). RT-PCR reagents were purchased from Promega (Madison, WI). Polynucleotide kinase was purchased from Pharmacia (Piscataway, NJ). 

The aim of this study is to elucidate whether camosic acid, camosol, rosmarinic acid and ursolic acid have any effects on apoptotic induction, since ursolic acid was previously reported to possess this effect previously (28). We used HL-60 cells to investigate the molecular mechanisms involved. We examined the effects of these rosemary phytochemicals on DNA fragmentation, activation of caspases, altering the mitochondrial function, ROS generation and releasing of cytochrome c from mitochondria. In the present study, we have demonstrated camosic acid, camosol, and ursolic acid induced apoptosis in HL-60 ceils and activated caspase-3 and caspase-9 via provoking the release of cytochrome c. 

Camosol, camosic acid, rosmarinic acid and ursolic acid were isolated from rosemary (/). Their sfructures are shovm in Figure 1. RNase A, proteinase K and propidium iodide were purchased from Sigma (St. Louis, MO). Substrates for caspase-1, Ac-YVAD-AMC casppase-3, Ac-DEVD-AMC caspase-8, Ac-lETD-AMC and caspase-9, Ac-LEHD-AMC were obtained from AnaSpec Inc. (San Jose, CA). DiOC6 (5) and DCFH-DA were obtained from Molecular Probes (Eugene, OR). 

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