AY-9944 inhibition of sterol biosynthesis inChlorella emersonii

WhenChlorella emersonii, a green alga, was cultured in the presence of 20 ppm AY-9944, a number of sterols accumulated which appear to be intermediates of sterol biosynthesis in this organism. The sterols isolated include 14α-methyl-ergost-8-en-3β-ol, 14α-methyl 24S-stigmast-8-en-3β-ol, 14α-methyl ergosta-8,24(28)-dien-3β-ol and 4α, 14α-dimethyl 24S-stigmast-8-en-3β-ol. Smaller quantities of several other sterols were found in addition to the normally occurring Δ⁷, chondrillasterol and Δ⁷. Control cultures were found to contain, in addition to the normally occurring sterols, smaller quantities of most of the sterols isolated from AY-9944 inhibited cultures. AY-9944 is a specific inhibitor of Δ⁷ in cholesterol biosynthesis in animals. However, sinceC. emersonii terminates sterol biosynthesis one step prior to the Δ⁷ step, AY-9944 apparently inhibits sterol biosynthesis prior to this step in this organism. The accumulation of 14α-methyl sterols in treated cultures suggests that AY-9944 is an effective inhibitor of the 14α-methyl removal inC. emersonii. Scientific Article No. A1865, Contribution No. 4775 of the Maryland Agricultural Experiment Station.     

Inhibition of sterol biosynthesis in chlorella ellipsoidea by AY-99441

WhenChlorella ellipsoidea was grown in the presence of 4 ppm AY-9944, complete inhibition of Δ⁵-sterol biosynthesis was achieved. However total sterol production remained unaltered. As a result a number of sterols accumulated that appear to be intermediates in sterol biosynthesis. These sterols were described and identified as (24S)-5α-ergost-8(9)-en3β-ol, (24S)-5α-stigmast-8(9)-en-3β-ol, 4α-methyl-(24S)-5α-ergosta-8, 14-dien-3β-ol, 4α-methyl-(24S)-5α-stigmasta-8, 14-dien-3β-ol, 4α-methyl-(24S)-5α-ergost-8(9)-en-3β-ol and (24S)-4α-methyl-5α-stigmast-8(9)-en-3β-ol. The occurrence of these sterols inChlorella ellipsoidea is the first time they have been noted in biological material. The accumulation of these sterols in treated cultures indicates that AY-9944 is an extremely effective inhibitor of the Δ⁸→Δ⁷ isomerase and the Δ¹⁴ reductase of these plants. The occurrence of small amounts of other sterols in treated cultures has led to a proposed pathway for thebiosynthesis of sterols inChlorella ellipsoidea. Scientific Article No. A1775, Contribution No. 4565 of the Maryland Agricultural Experiment Station.     

Synthesis of Δ5,22-cholestadien-3β-ol from Δ5,7,22-cholestatrien-3β-ol by a liver enzyme

The rat liver enzyme system, which catalyzes reduction of Δ^5,7,24-cholestatrien-3β-ol to cholesterol (Δ⁵-cholesten-3β-ol), converted radiolabeled Δ^5,7,22-cholestatrien-3β-ol to Δ^5,22-cholestadien-3β-ol, but not to cholesterol. This enzyme system thus contains membrane-bound Δ⁷ and Δ²⁴-reductases and no Δ²²-reductase. Kinetic and competition studies showed that the enzymes system contains a single Δ^5,7-sterol Δ⁷-reductase, which is not influenced by unsaturtion at the Δ²²-position of the sterol side chain. The identity of Δ^5,22-cholestadienol was established by chromatographic, spectral and chemical analyses. Use of the enzyme system and readily available Δ^5,7,22-cholestatrienol provides a facile procedure for specific production of Δ^5,22-cholestradien-3β-ol in quantity.     

The lipids of the common house cricket,Acheta domesticus L. III. Sterols

Sterols constitute 1.95% of the total extractable lipids ofAcheta domesticus L., of which 18% are esterified. The free sterols consist of cholestane-3β-ol (0.5%), Δ⁵-cholestene-3β-ol (83.5%), Δ⁷-cholestene-3β-ol (2.3%) Δ^5,7-cholestadiene-3β-ol (3%), Δ^5,22-cholestadiene-3β-ol (4%), Δ^5,7,22-cholestatriene-3β-ol (0.2%), campestane-3β-ol (0.03%), Δ⁵-campestene-3β-ol (1.0%), Δ⁷-campestene-3β-ol (trace), Δ^5,7-campestadiene-3β-ol (0.2%), stigmastane-3β-ol (0.09%), Δ⁵-stigmastene-3β-ol (2.1%), Δ⁷-stigmastene-3β-ol (0.04%), Δ^5,7-stigmastadiene-3β-ol (0.4%), Δ^5,22-stigmastadiene-3βol (0.1%). The same sterols are present in the esterified sterol fraction. Δ⁷-Sterols and Δ^5,7-sterols are present in significantly larger amounts in the esterified fraction than in the free sterol fraction. By a comparison with the sterols of the cricket food, it is clear thatA. domesticus is capable of removing methyl and ethyl groups from C-24 of sterols of the campestane and stigmastane type. The ability to introduce a Δ⁷ double bond into saturated and Δ⁵-sterols is indicated, and it is suggested that Δ⁷-sterols of the C₂₇, C₂₈, and C₂₉ sterol series may be intermediates in the conversion of Δ⁵-sterols to Δ^5,7-sterols. Associate Professor, Department of Chemistry, University of Michigan, Ann Arbor, Mich.; Alfred P. Sloan Foundation Fellow, 1968–68. Public Health Service Predoctoral Fellow, 1968–67.     

Inhibitors of cholesterol synthesis and myelin formation

Inhibitors of cholesterol biosynthesis AY 9944 and 20,25 diazacholesterol were administered by stomach tube to suckling rats in varying doses during the time of rapid myelination (15–22 days of age). Purified myelin was prepared from the brains and spinal cords, and the sterol content analyzed. Up to 50% of the myelin sterol consisted of desmosterol in rats treated with 20,25 diazacholesterol, while 7-dehydrocholesterol comprised at least one third of the myelin sterol in rats administered AY 9944. Myelin from rats treated with both compounds contained desmosterol, 7-dehydrocholesterol, Δ^5,7,24 cholestatriene-3-β-ol and an unknown sterol, the four comprising about 45% of the total sterol. The proportion of phospholipid: galactolipid-total sterol in myelin from the drug-treated rats was not significantly different from the normal, although much less myelin was recovered. Brain and spinal cord slices from 22 to 25-day rats pretreated with inhibitors showed decreased uptake of U-¹⁴C-glucose into all myelin components. The decreased uptake was approximately proportional in all lipids and the protein was also affected. It is proposed that myelin composition is fixed, and that a deficiency of one of the lipid components will limit the assembly of the whole lipid portion of the membrane. Presented at the AOCS Meeting, San Francisco, April 1969.     

Nature of fecal sterols and intestinal bacterial flora

Sterol excretion in the spontaneously atherosclerosis-susceptible White Carneau (WC) pigeon, the Silver King (SK) pigeon and the Show Racer (SR) pigeon was studied by thin layer chromatography (TLC), argentation TLC and gas liquid chromatography. Unlike man and the chicken, these pigeons excreted no coprostanol or coprostanone derivatives of sterols. Moreover incubation of¹⁴C-labeled cholesterol with pigeon feces indicated that, also unlike man and the chicken, these pigeons are unable to convert it to coprostanol. Bacteriologic examination revealed the absence of gram-negative anaerobic flora and of members of the genusBifidobacterium in both the WC and SR pigeons. On the other hand, one of the two SK pigeons examined showed evidence of the presence of bothBacteroids fragilis andB. bifidum in the upper intestinal tract. Although no qualitative experiments were performed, no unusual characteristics of the aerobic flora were noted in these pigeons. In addition, analysis of human stool specimens indicated a “normal” bowel flora. The flora of the intestinal tract of the chicken is similar to that of the human. Because of this similarity, it appears that differences in environment (living conditions, diets) between the human and the chicken are of little consequence. The results obtained in this study suggest the possibility that the anaerobic gram-negative flora and sponsible, at least in part, for the chemical conversion of cholesterol to coprostanol. Presented in part at the Fourth International Symposium on Drugs Affecting Lipid Metabolism, Philadelphia, September 1971, and at the AOCS Meeting, Los Angeles, April 1972. The following nomenclature has been used for the steroids referred to in this paper: cholesterol, cholest-Δ⁵-en-3β-ol; coprostanol, 5β-cholestan-3β-ol; campesterol, 24-methylcholest-Δ⁵-en-3βol; stigmasterol, 24-ethylcholest-Δ^5,22-dien-3β-ol; β-sitosterol, 24-ethylcholest-Δ⁵-en-3β-ol; coprocampestanol, 24-methyl-5β-cholestan-3β-ol; coprostigmastenol, 24-ethyl-5β-cholest-Δ²²-en-3β-ol; coprostigmastanol, 24-ethyl-5β-cholestan-3β-ol; coprostanone, 5β-cholestan-3-one; campestanone, 24-methyl-5β-cholestan-3-one; stigmastenone, 24-ethyl-5β-cholest-Δ²²-en-3-one; and β-sitostanone, 24-ethyl-5β-cholestan-3-one.     

Purification of campesterol and preparation of 7-dehydrocampesterol, 7-campestenol and campestanol

Campesterol ([24R]-24-methyl-5-cholesten-3β-ol) was isolated from a soybean sterol mixture and purified on a small scale by column chromatography and on a large scale by crystallization from acetone to 97% purity. From this, 7-dehydrocampesterol ([24R]-24-methyl-5,7-cholestadien-3β-ol) and campestanol ([24R]-24-methyl-cholestan-3β-ol) were prepared. The acetates and benzoates of all four compounds also were obtained and compared to the corresponding members of the ergostane series. Contribution 2189, Arizona Agricultural Experiment Station.     

Anatomical distribution of sterols in oysters (Crassostrea gigas)

Oysters (Crassostrea gigas) contain at least 8 predominant sterols as determined by gas liquid chromatography and a modified Liebermann-Burchard reaction. These sterols and the average amount found in mg/100 are: C₂₆-sterol (22-trans-24-norcholesta-5, 22-diene-3β-ol), 19.1; 22-dehydrocholesterol, 15.1; cholesterol, 46.8; brassicasterol, 27.2; Δ^5,7-sterols (i.e., 7-dehydrocholesterol) 22.5; 24-methylenecholesterol 29.1; 24-ethylcholesta-5,22-diene-3β-ol, 1.2; and 24-ethylcholesta-5-en-3β-ol, 12.7. The distribution of these sterols appears uniform (r²=0.938) between 5 major organs of the oyster. The percent body mass vs percent total sterols in these 5 organs are: mantle 44.1–41.4; visceral mass 30.3–36.7; gills 13.2–11.7; adductor muscle 8.3–3.7; and labial palps 4.2–6.5. The possible sources of these sterols are discussed.     

Cooccurrence of C-24 alkylated Δ7- and Δ5-Sterols in the leaves ofBeta vulgarisin the leaves ofBeta vulgaris

The 4-desmethylsterols from the leaves ofBeta vulgaris are a mixture of Δ⁷-sterols (71%) and Δ⁵-sterols (29%). The Δ⁷-sterols isolated are spinasterol (24α-ethylcholesta-7,22-dien-3β-ol; 45%), 22-dihydrospinasterol (24α-ethylcholest-7-en-3β-ol; 24%), and avenasterol (24-ethylcholesta-7,24(28)-dien-3β-ol; 1.5%). The Δ⁵-sterols isolated are sitosterol (24α-ethylcholest-5-en-3β-ol; 15%), 24ζ-ethylcholesta-5,22-dien-3β-ol (7.5%), and 24ζ-methylcholest-5-en-3β-ol (7%).     

Unusual tetraene sterols in some phytoplankton

Sterols were analyzed from four phytoplankton strains which are under investigation as possible sources of food for oysters in culture. One strain ofPyramimonas contained only 24-methylenecholesterol as a major sterol component.Pyramimonas grossii, Chlorella autotrophica andDunaliella tertiolecta each contained a complex mixture of C₂₈ and C₂₉ sterols with Δ⁷, Δ^5,7 and Δ^5,7,9(11) nuclear double bond systems. Sterols were found both with and without the C-22 side chain double bond. Ergosterol and 7-dehydroporiferasterol were the principal sterols in each of the latter three species, which also contained the rare tetraene sterols, 24-methylcholesta-5,7,9(11),22-tetraen-3β-ol and 24-ethylcholesta-5,7,9(11),22-tetraen-3β-ol.     

Diversity of sterol composition in the family chenopodiaceae

The predominant 4-desmethylsterols from the leaves of 13 species in eight genera of the family Chenopodiaceae are 24α-ethylsterols. In four species,Chenopodium ambrosioides L.,C. rubrum L.,Salicornia europaea L. andS. bigelovii Torr., the C-22(23) double bond is introduced into more than 70% of the 24α-ethylsterols producing spinasterol (24α-ethylcholesta-7,22E-dien-3β-ol) in the first two species and mixtures of spinasterol and stigmasterol (24α-ethylcholesta-5,22E-dien-3β-ol) in the latter species. The saturated side chain analogues predominate with more than 70% of the 24α-ethylsterols in eight species.Salsola kali L.,Suaeda linearis (Ell.) Moq.,Kochia scoparia (L.) Roth., andBassia hirsute (L.) Aschers. synthesize sitosterol (24α-ethylcholest-5-en-3β-ol), andAtriplex arenaria Nutt.,C. album L.,C. urbicum L. andC. leptophyllum Nutt. possess mixtures of sitosterol and 22-dihydrospinasterol (24α-ethylcholest-7-en-3β-ol). Sitostanol (24α-ethyl-5α-cholestan-3β-ol) was isolated fromSuaeda linearis as an 18% component of the total 4-desmethylsterol and in lesser amounts from four other species. In all species synthesizing 24-ethyl-Δ⁵-sterols, a 24ξ-methylcholest-5-en-3β-ol was also present at 1.0–20% of the total 4-desmethylsterol. Avenasterol [24-ethylcholesta-7,24(28)Z-dien-3β-ol], isofucosterol [24-ethylcholesta-5,24(28)Z-dien-3β-ol), cholesterol (cholest-5-en-3β-ol) and 24ξ-methyl-5α-cholestan-3β-ol also were isolated from several species. Species in the family Chenopodiaceae and the type genusChenopodium may be categorized into one of three groups based on sterol biosynthesis: the Δ⁷-sterol producers; the Δ⁵-sterol producers, and those producing mixtures of both Δ⁷- and Δ⁵-sterols in relatively fixed percentage compositions.     

Metabolism of sitosteryl β-D-glucoside and its nutritional effects in rats

[4-¹⁴C]Sitosteryl β-D-glucoside, intragastrically administered to rats, was not absorbed by the intestinal mucosa. At three hr after the application, radioactivity was concentrated almost exclusively in the digesta of stomach, small intestine as well as cecum and colon, whereas only low proportions of radioactively labeled compounds were found in the various tissues of the gastrointestinal tract. Minor proportions of labeled metabolites of [4-¹⁴C]sitosteryl β-D-glucoside, such as sitosterol and sitosteryl esters, were formed in the small intestine in vivo and in slices of small intestine in vitro. In the tissues of cecum and colon as well as the digesta derived from them, high proportions of labeled coprositostanol, i.e. 24α-ethyl-5β-cholestan-3β-ol, that obviously had been formed by bacterial degradation of the substrate were detected. The feeding of sitosteryl β-D-glucoside (0.5 g/kg body weight×day) over a period of four weeks did not alter significantly body weights or organ weights of rats. Analyses of steryl lipids of the various organs and tissues confirmed the findings obtained with the radioactive substrate: neither sitosteryl β-D-glucoside nor sitosterol or sitosteryl esters derived therefrom had been transported in appreciable amounts to organs and tissues outside the alimentary canal during the feeding period. Minor proportions of unmetabolized sitosteryl β-D-glucoside were detected in the tissues of stomach and intestine, whereas large proportions of the substrate were found in feces of rats that had received the sitosteryl β-D-glucoside-containing diet; coprositostanol was found in feces of these animals in high proportions as well. Thus, the use of sitosteryl β-D-glucoside as emulsifier or preservative in food and feed does not appear to involve any risk. The systematic nomenclature of the sterols referred to by trivial names is, cholest-5-en-3β-ol (cholesterol); 5α-cholestan-3β-ol (5α-cholestanol); 5β-cholestan-3β-ol (5β-cholestanol, coprostanol); 24α-methylcholest-5-en-3β-ol (campesterol); 24α-methyl-5α-cholestan-3β-ol (5α-campestanol); 24α-methyl-5β-cholestan-3β-ol (5β-campestanol, coprocampestanol); 24α-methyl-cholesta-5,22-dien-3β-ol (brassicasterol); 24α-ethylcholest-5-en-3β-ol (sitosterol, β-sitosterol); 24α-ethyl-5α-cholestan-3β-ol (5α-sitostanol); 24α-ethyl-5β-cholestan-3β-cholestan-3β-ol (5β-sitostanol, coprositostanol); 24α-ethylcholesta-5,22-dien-3β-ol (stigmasterol).     

Sterols of cucurbitaceae: The configurations at C-24 of 24-Alkyl-Δ5-,Δ7- and Δ8-sterols

The major sterols of the seeds ofBenincasa cerifera, Cucumis sativus, Cucurbita maxima, C. pepo andTrichosanthes japonica and of the mature plant tissues (leaves and stems) ofCitrullus battich, Cucumis sativus andGynostemma pentaphyllum of the family Cucurbitaceae were 24-ethyl-Δ⁷-sterols which were accompanied by small amounts of saturated and Δ⁵-and Δ⁸-sterols. The 24-ethyl-Δ^7,22,Δ^7,25(27) and Δ^7,22,25(27)-sterols constituted the predominant sterols for the seed materials, whereas the 24-ethyl-Δ⁷ and Δ^7,22-sterols were the major ones for the mature plant tissues. The configurations of C-24 of the alkylsterols were examined by high resolution¹H NMR and¹³C NMR spectroscopy. Most of the 24-methyl- and 24-ethylsterols examined which lack a Δ²⁵⁽²⁷⁾-bond (i.e., 24-methyl-, 24-methyl-Δ²²-, 24-ethyl- and 24-ethyl-Δ²² sterols) were shown to occur as the C-24 epimeric mixtures in which the 24α-epimers predominated in most cases. The 24-ethylsterols which possess a Δ²⁵⁽²⁷⁾ (i.e., 24-ethyl-Δ²⁵⁽²⁷⁾-and 24-ethyl-Δ^7,22,25(27)-sterols) were, on the other hand, composed of only 24β-epimers. The Δ⁸-sterols identified and characterized were four 24-ethyl-sterols: 24α-and 24β-ethyl-5α-cholesta-8,22-dien-3β-ol, 24β-ethyl-5α-cholesta-8,25(27)-dien-3β-ol and 24β-ethyl-5α-cholesta-8,22,25(27)-trien-3β-ol. This seems to be the first case of the detection of Δ⁸-sterols lacking a 4-methyl group in higher plants, and among the four Δ⁸-sterols the latter two are considered to be new sterols. The probable biogenetic role of the Δ⁸-sterols and the possible biosynthetic pathways leading to the 24α- and 24β-alkylsterols in Cucurbitaceae are discussed.     

Desmosterol in developing rat brain

The brain of the young rat contains appreciable amounts of desmosterol (24-dehydrocholesterol). The peak desmosterol concentration is seen during the first week of life and only traces of this sterol are found at 21 days. The spinal cord also contains some desmosterol. Rat brain desmosterol is distributed in the white matter, gray matter and cerebellum and occurs in the same proportion to cholesterol in each of these brain fractions. Rat brain contains a small amount of sterol ester but no appreciable amounts of desmosterol are present in this fraction. Studies carried out in intact animals injected either intraperitoneally or intracerebrally with mevalonic acid-2-¹⁴C or glucose-U-¹⁴C indicate the biosynthetic origin or brain desmosterol. Rat brain slices (1–20 day old) incubated in suitably fortified medium convert sodium acetate-2-¹⁴C and glucose-U-¹⁴C to desmosterol, whereas brain slices from adult rats yielded no radioactive desmosterol under similar conditions. When labeled desmosterol was incubated with young rat brain slices, it was converted to cholesterol. When pregnant rats were treated with triparanol (20 mg/kg/day) during the course of their pregnancy, they either resorbed the fetuses or gave birth to small, stillborn litters. The brains of the progeny of triparanol treated mothers contained large amounts of desmosterol as well as another sterol which may be Δ^7,24-cholestadiene-3β-ol.     

Dominance of Δ7-sterols in the family caryophyllaceaein the family caryophyllaceae

The predominant 4-desmethylsterols from the leaves of 12 species in 11 genera of the family Caryophyllaceae are 24-ethyl-Δ⁷-sterols. In eight species,Scleranthus annus L.,Paronychia virginica Spreng.,Lychnis alba Mill.,Silene cucubalus Wibel,Dianthus armeria L.,Gypsophilia paniculata L.,Saponaria officinales L. andMyosoton aquaticum (L.) Moench, the major sterols are spinasterol (24α-ethylcholesta-7,22E-dien-3β-ol) and 22-dihydrospinasterol (24α-ethylcholest-7-en-3β-ol), with spinasterol at more than 60% of the desmethylsterol in the latter six species. Both 24α-and 24β-ethyl-Δ⁷-sterols are present in two species,Minuartia caroliniana Walt. andSpergula arvensis L., which possess 24β-ethylcholesta-7,25(27)-dien-3β-ol and 24β-ethylcholesta-7,22E,25(27)-trien-3β-ol as well as spinasterol and 22-dihydrospinasterol.Cerastium arvense L.,C. vulgatum L. andArenaria serpyllifolia L. possess 24-alkyl-Δ⁵ and Δ⁷-sterols. These three species synthesize sitosterol (24α-ethylcholest-5-en-3β-ol), 24ζ-methylcholest-5-en-3β-ol, spinasterol, 22-dihydrospinasterol and the stanols, sitostanol (24α-ethyl-5α-cholestan-3β-ol) and 24ζ-methyl-5α-cholestan-3β-ol. Avenasterol (24-ethylcholesta-7,24(28)Z-dien-3β-ol) was also isolated from five species. Sterol biosynthetic capability may be a useful characteristic in examining the taxonomic relatedness of plants in the Caryophyllaceae.     

The effect of AY-9944 on yeast sterol and sterol ester metabolism

The effects of the hypocholesterolemic drug AY-9944 (trans-1,4-bis(2-chlorobenzylaminoethyl)cyclohexane dihydrochloride) at two concentrations (10⁻⁴M and 5×10⁻⁴M), on the synthesis of sterols and sterol esters bySaccharomyces cerevisiae were investigated. Although growth was not markedly affected by the drug, there was a decrease in the free sterol to sterol ester ratio with increased drug concentration. A concomitant increase in the saturated fatty acids esterified to sterol relative to the unsaturated fatty acids was also noted in response to increased drug concentration. Ergosterol accounted for 94.7% of the free sterol in the control culture and for 87.8% of the 5×10⁻⁴M drug-treated culture, respectively. However, in the sterol ester fraction, the ergosterol content decreased from a value of 45.1% in the control culture to 2.4% in the 5×10⁻⁴M AY-9944 treated culture. The sterol ester fraction simultaneously showed increased levels of the Δ⁸ sterol, fecosterol, in response to increased drug concentration from a 7.4% control value to 57.4% in the 5×10⁻⁴M drug-treated culture. The accumulation of the Δ⁸ sterol suggests that the site of action of the drug is probably at the Δ⁸ to Δ⁷ isomerase step in the biosynthesis of ergosterol. The fact that ergosterol is retained as the major free sterol suggests a biological advantage to the retention of this particular sterol. In addition, the near normal growth in the presence of the drug, in spite of the occurrence of an altered sterol ester profile, indicates that the composition of the sterol ester fraction is not as critical as the free sterol fraction. These results form a portion of a dissertation submitted to the Graduate School, University of Maryland, College Park, MD, by Ravi Pereira, in partial fulfillment of the requirements for the Ph.D. degree in Biochemistry.     

Accumulation of ergosta-8,14-dien-3β-ol bySaccharomyces cerevisiae cultured with an azasterol antimycotic agent

15-Aza-24-methylene-D-homocholesta-8,14-dien-3β-ol, an antimycotic agent, at a concentration of 75 ng/ml inhibited ergosterol biosynthesis inSaccharomyces cerevisiae strain 3701 B resulting in the accumulation of an unusual sterol. Experimental data presented indicate that this sterol is ergosta-8,14-dien-3β-ol. The accumulation of the compound is supportive of current models of biosynthetic pathways for sterols in yeast and is consistent with inhibition by the azasterol of the Δ¹⁴ sterol reductase.     

Sterols ofKalanchoe pinnata: First report of the isolation of both C-24 epimers of 24-alkyl-Δ25-sterols from a higher plant

Eighteen sterols were isolated from the aerial parts ofKalanchoe pinnata (Crassulaceae) including four novel sterols,viz. (24R)-stigmasta-5,25-dien-3β-ol (24-epiclerosterol), (24R)-5α-stigmasta-7,25-dien-3β-ol, 5α-stigmast-24-en-3β-ol, and 25-methyl-5α-ergost-24(28)-en-3β-ol. 24-Epiclerosterol and its Δ⁷-analog occur together with their 24S/β-epimers in the same plant making this the first report of the isolation of both C-24 epimers of Δ²⁵-unsaturated 24-alkylsterols from a non-marine organism. Iodine-catalyzed isomerization of stigmasta-5,24-dien-3β-ol (24-ethyldesmosterol), the main sterol ofK. pinnata, yielded 24-epiclerosterol among other products.     

The sterols ofCucurbita moschata (“calabacita”) seed oil

From the sterol fraction of seed oil from commercialCucurbita moschata Dutch (“calabacita”) Δ⁵ and Δ⁷ sterols having saturated and unsaturated side chain were isolated by chromatographic procedures and characterized by spectroscopic (¹H and¹³C-nuclear magnetic resonance, mass spectrometry) methods. The main components were identified as 24S-ethyl 5α-cholesta-7,22E-dien-3β-ol (α-spinasterol); 24S-ethyl 5α-cholesta-7,22E, 25-trien-3β-ol (25-dehydrochondrillasterol); 24S-ethyl 5α-cholesta-7, 25-dien-3β-ol; 24R-ethylcholesta-7-en-3β-ol (Δ⁷-stigmastenol) and 24-ethyl-cholesta-7, 24(28)-dien-3β-ol (Δ^7,24(28)-stigmastadienol).Lipids 31, 1205–1208 (1996).     

Conversion of cyclolaudenol to 24α- and 24β-ethylsterols in the cucurbitaceae

[2-³H]Cyclolaudenol was converted into α-spinasterol, 24β-ethylcholesta-7,25-dien-3β-ol and 24β-ethylcholesta-7,22,25-trien-3β-ol by seedlings ofCucurbita maxima. As 24-methylenecycloartanol is the obligatory precursor of 24-ethylsterols, it can be assumed that the transformation of cyclolaudenol to 24-methylenecycloartanol must have occurred. These results lead us to postulate the existence, in the Cucurbitaceae family, of an enzymatic system capable of isomerizing Δ²⁵ alkylsterols into Δ²⁵⁽²⁸⁾ sterols.