Determination of norgestimate and ethinyl estradiol in tablets by high-performance liquid chromatography

A rapid, simple, stability-indicating assay procedure for norgestimate [(+)-13-ethyl-17-hydroxy-18,19-dinor-17 alpha-pregn-4-en-20-yn-3-one oxime acetate], a new progestational agent, and ethinyl estradiol (19-nor-17 alpha-pregna-1,3,5(10)-trien-20-yne-3,17-diol) in single- and composite-tablet analyses was developed using high-performance liquid chromatography. Norgestimate and ethinyl estradiol were extracted from the tablet matrix with methanol containing an internal standard. An aliquot was chromatographed on a 5-microns, reversed-phase column using water:tetrahydrofuran:methanol solution (65:25:10 v/v/v) as the mobile phase. The selectivity of the chromatographic system for intact norgestimate and ethinyl estradiol was demonstrated by resolving both compounds from various potential degradation products of each compound. An essential property of the chromatographic system was its ability to separate norgestimate as its syn and anti isomers. The method is linear, quantitative, and reproducible.     

Methandrostenolone: metabolism in the rabbit

Methandrostenolone and the fully reduced metabolites 17 alpha-methyl-5 alpha-androstane-3 beta, 17 beta-diol and 17 alpha-methyl-5 beta-androstane-3 alpha, 17 beta-diol, the partially reduced and hydroxylated metabolites 16 alpha, 17 beta-dihydroxy-17 alpha-methyl-5 beta-androst-1-en-3-one and 16 beta, 17 beta-dihydroxy-17 alpha-methyl-5 beta-androst-1-en-3-one, the monohydroxylated metabolites 6 beta, 17 beta-dihydroxy-17 alpha-methyl-1,4-androstadien-3-one and 16 beta, 17 beta-dihydroxy-17 alpha-methyl-1,4-androstadien-3-one, and the dihydroxylated metabolite 6 beta, 16 beta, 17 beta-trihydroxy-17 beta-trihydroxy-17 alpha-methyl-1,4-androstadien-3-one have been isolated and identified in the urine of rabbits orally dosed with methandrostenolone. C-16 Hydroxylated and dihydroxylated metabolites have not been previously reported from methandrostenolone. No evidence for epimerization at the C-17 position was observed in the rabbit.     

New monoterpenes from Mentha microphylla

Four new monoterpenes, (1 R*,2 S*)-1,2-dihydroxy- p-menth-4(8)-en-3-one ( 1), (4 S*)-4-hydroxy- p-mentha-1,8-dien-3-one ( 2), (4 S*)-4-methoxy- p-mentha-1,8-dien-3-one ( 3), and (3 R*,4 R*,6 S*)-3-acetoxy-6-hydroxy- p-mentha-1,8-diene ( 4) together with four known compounds have been isolated from the aerial parts of Mentha microphylla. The structures were determined using spectroscopic methods particularly high resolution (1)H-, (13)C-NMR as well as 2D (1)H- (1)H COSY, HMQC and HMBC analysis.     

6-hydroxy-4-en-3-one sterols from the marine sponge Iotrochoto birotulata

Five sterols with a nucleus skeleton of 6-hydroxy-4-en-3-one, namely cholesta-6beta-hydroxy-4-en-3-one (1), ergosta-6beta-hydroxy-4,24(28)-dien-3-one (2), ergosta-6alpha-hydroxy-4,24(28)-dien-3-one (3), ergosta-6alpha-hydroxy-4,22-E-dien-3-one (4), and ergosta-28-methyl-6beta-hydroxy-4,24(28)-dien-3-one (5) have been isolated from the marine sponge Iotrochoto birotulata, collected from the southern China sea. Sterols 2-4 are new compounds, and 1 has been isolated from marine organisms for the first time. The structures have been elucidated on the basis of extensive spectroscopic and chemical properties.     

Isolation and identification of novel impurities in spironolactone

Three known and five new steroidal compounds as impurities in spironolactone were isolated from the enriched mother liquor by using various chromatographic methods. Their structures were elucidated by spectrometric analysis. New compounds were characterized as 3-(3,3-dimethoxy-5,7-epidithio-17β-hydroxy-4-androstan-17-yl) propionic acid γ-lactone (6); 3-(3-oxo-7-acetylthio-6β,17β-dihydroxy-4-androsten-17-yl) propionic acid γ-lactone (7); 7-acetylthio-17β-20-isopropylidendioxy-21-nor-17-pregn-4-en-3-one (8); 7-acetylthio-3-oxo-pregna-4,17(20)i-dien-22-oic acid methyl ester (9) and 7-acetylthio-17-methyl-18-nor-androsta-4-en-3-one (10).     

Novel polyketide metabolites from Streptomyces rimosus mutant strain R1059

Three novel polyketide metabolites were isolated from laboratory-scale fermentation of the Streptomyces rimosus mutant strain R1059. Structural elucidation of the compounds was based on NMR experiments. The compounds were characterized as naphthalene derivatives: (rel)-4beta,8-dihydroxy-3alpha-hydroxymethyl-4alpha-methyl-1,2,3,4-tetrahydronaphthalene1-one (1), 4xi8-dihydroxy-3-hydroxymethyl-4xi-methyl-1,4-dihydronaphthalene-1-one (2) and (rel)-4beta,8-dihydroxy-3alpha-O-[alpha-glucopyranosyl]hydroxymethyl-4alpha-methyl-1,2,3,4-tetrahydronaphthalene-1-one (3). The compounds isolated appear to be derived via a shorter polyketide backbone than oxytetracycline (4), the normal end-product made by the parent of this strain. Compound 3 was the glucoside of 1 and must be formed as a post-PKS reaction by the activation of a glycosyl transferase, which has not been reported in this species before.     

Biotransformation of 1,4-cineole, a monoterpene ether

1. The metabolism of 1,4-cineole, a monoterpene ether, was studied in the rabbit. 2. Four neutral and one acidic metabolites were isolated from the urine and shown to be 9-hydroxy-1,4-cineole, 3,8-dihydroxy-1,4-cineole, 8,9-dihydroxy-1,4-cineole, 1,4-cineole-8-en-9-ol and 1,4-cineole-9-carboxylic acid.     

Biotransformation of 1,4-Cineole,a Monoterpene Ether

1.The metabolism of 1,4-cineole, a monoterpene ether, was studied in the rabbit.

2.Four neutral and one acidic metabolites were isolated from the urine and shown to be 9-hydroxy-1,4-cineole, 3,8-dihydroxy-1,4-cineole, 8,9-dihydroxy-1,4-cineole, 1,4-cineole-8-en-9-ol and 1,4-cineole-9-carboxylic acid.     

Androstene-17-thioketals. 1st communication: glucocorticoid receptor binding, antiproliferative and antiinflammatory activities of some novel 20-thiasteroids (androstene-17-thioketals)

The unique replacements of the alpha-hydroxyl and beta-ketol groups of corticoids at C17 with selected, simple alkylthio or (2-fluoroalkyl)thio groups resulted in the structurally novel steroids, C17-alkylthioketals of 9 alpha-fluoro-11 beta-hydroxy-androsta-1,4-diene-3,17-dione. The described androstene-17-thioketals (20-thiasteroids) had high affinities for the glucocorticoid receptor protein of rat liver cytosol. Most were more potent than triamcinolone acetonide, a clinically moderately potent corticoid, in antiproliferative and antiinflammatory activities in mice. Specifically, (11 beta, 17 alpha)-17-(ethylthio)-9 alpha-fluoro-11 beta-hydroxy-17-(methylthio) androsta-1,4-dien-3-one (tipredane, SQ 27,239) and (11 beta, 17 alpha)-17-(ethylthio)-9 alpha-fluoro-17-[2-(fluoroethyl)thio] - 11 beta - hydroxy-androsta-1,4-dien-3-one (SQ 28,300), topically applied, were as potent as halcinonide, a clinically highly potent corticoid, in inhibition of croton oil-induced edema in the mouse. It is suggested that both thiasteroids could be moderately to highly potent topical antiinflammatory agents in man.     

Oxidation of furans. 3. Estrogenic properties of lactones and anhydrides derived from the oxidation of 17alpha-[3-furyl]estradiol derivatives.

The oxidation of 17alpha-[3-furyl]estradiol derivatives and the estrogenic properties of the resulting isomeric 17,21-and 17,23-dihydroxy-19,24-dinor-17alpha-chola-1,3,5(10)20(22)-tetraenoic acid(20leads to 23) gamma-lactones as well as those of the related 17-hydroxy-19-nor-17alpha-pregna-1,3,5(10)-triene-20,21-dicarboxylic acid anhydrides and gamma-lactones are described. Of these, only lactones 3c,e,h and 5b retained the same degree and profile of estrogenic activity as the starting 17alpha-[3-furyl]estradiols.     

Steroids from Harrisonia abyssinica

In addition to the known sterols and ketosteroids beta-sitosterol (24 alpha-ethylcholest-5-en-3 beta-ol), stigmasterol (24 alpha-ethylcholesta-5,22-dien-3 beta-ol), campesterol (24 alpha-methylcholest-5-en-3 beta-ol), beta-sitostenone (stigmast-4-en-3-one, 24 alpha-ethylcholest-4-en-3-one), stigmastenone (stigmasta-4,22-dien-3-one, 24 alpha-ethylcholesta-4,22-dien-3-one), campestenone (24 alpha-methylcholest-4-en-3-one), and stigmasta-3,5-dien-7-one (24 alpha-ethylcholesta-3,5-dien-7-one), the new steroids stigmasta-3,5,22-trien-7-one (24 alpha-ethylcholesta-3,5,22-trien-7-one), and campesta-3,5-dien-7-one (24 alpha-methylcholesta-3,5-dien-7-one) were isolated from the stem bark of Harrisonia abyssinica and identified by NMR and mass spectrometry.     

Pregnane glycosides from Solanum nigrum

Two new pregnane saponins, solanigroside A (1) and solanigroside B (2), along with two known compounds (3 and 4), were isolated from 60% ethanolic extract of the dried herb of Solanum nigrum L. The structures of 1 and 2 were elucidated as 5alpha-pregn-16-en-3beta-ol-20-one 3-O-beta-D-xylopyranosyl-(1 --> 3)-O-[alpha-L-arabinopyranosyl-(1 --> 2)]-O-beta-D-glucopyranosyl-(1 --> 4)-O-[alpha-L-rhamnopyranosyl-(1 --> 2)]-O-beta-D-galactopyranoside (1) and 5alpha-pregn-16-en-3beta-ol-20-one 3-O-beta-D-glucopyranosyl-(1 --> 2)-O-[beta-D-glucopyranosyl-(1 --> 3)]-O-beta-D-glucopyranosyl-(1 --> 4)-O-beta-D-galactopyranoside (2), respectively, on the basis of extensive spectroscopic analysis as well as comparison with reported spectroscopic data of related compounds. This paper deals with the isolation and structural characterisation of pregnane glycosides from S. nigrum L.     

Synthesis of 11 beta,13 beta- and 13 beta,16 beta-propano steroids: probes of hormonal activity.

Syntheses of 11 beta,13 beta- and 13 beta,16 beta-propano derivatives of 17 alpha-ethynyl-17 beta-hydroxygon-4-en-3-one are described. The 13 beta,16 beta bridge was constructed by intramolecular alkylation of the C-16 enolate anion from 3-methoxy-13 beta-[3'-(tosyloxy)propyl]gona-3,5-dien-17-one, the latter being obtained via Birch reduction of both aryl groups of 17 beta-hydroxy-3-methoxy-13 beta-(3'-phenoxypropyl)gona-1,3,5(10),8-tetraene (1). The 11 beta,13 beta bridge was constructed by Prins cyclization of 17 beta-acetoxy-3-methoxy-13 beta-(3'-oxopropyl)gona-1,3,5(10),9(11)-tetraene, itself obtained via Birch reduction of only the side-chain aryl group of 1. Binding affinities of certain of these compounds and substituted 13 beta-propyl derivatives of 17 alpha-ethynyl-17 beta-hydroxygon-4-en-3-one for the uterine cytosol receptor of progesterone are reported, and the origin of the high progestational activity of norgestrel and 11 beta-substituted progestins is discussed.     

Metabolism of a new synthetic progestagen,Org 2969, in female volunteers. Pharmacokinetics after an oral dose

Summary The pharmacokinetics of a new synthetic progestagen, Org 2969 (13-ethyl-11-methylene-18,19-dinor-17-pregn-4-en-20-yn-17-ol) and its presumed active metabolite, 3-keto-Org 2969, were studied in five healthy female volunteers. During the first part of the study (Phase I), four volunteers ingested as a single dose 50 µg (about 100 µCi) of [16-³H]-Org 2969 together with 50 µg of ethinyl oestradiol. During the second part (Phase II), ten days of pretreatment with non-readioactive Org 2969 and ethinyl oestradiol preceded dosing, which was similar to that in Phase I. The fifth volunteer ingested 2500 µg of Org 2969 as a single dose. The concentrations of Org 2969 and 3-keto-Org 2969 in serum were measured by specific radioimmunoassay, and by as radioactivity. The maximum serum level of Org 2969 of 0.2–0.3% of the dose/litre was obtained 0.8–1.3 h after administration, and it was followed by a mono-exponential decrease, the half-life being 1.3–1.5 hours. No difference in Org 2969 levels was seen between Phase I and Phase II studies. The maximum 3-keto-Org 2969 serum level in Phase I was 0.4–0.8% of the dose/litre, 1–3 h after administration. A two-compartment open body model adequately described the data. The half-life of elimination was 16 hours. There was a considerable change in the single dose kinetics between Phase I and Phase II in the case of 3-keto-Org 2969. In Phase II the maximum serum level was 2.0–3.4% of the dose/litre, and there was no significant change in half-life. The change was considered to be due to a decrease in the apparent volume of distribution as a result of an increased number of binding sites on sex hormone-binding globulin induced by ethinyl oestradiol during the pretreatment period, and/or to an increase in the fraction of Org 2969 metabolized to 3-keto-Org 2969. The two simultaneous processes contributed to the change in kinetics and to the production of a steady state level of 3-keto-Org 2969 which resulted in a steady state within the first 10 days of daily administration of Org 2969 and ethinyl oestradiol.The following codes and trivial names were used Org 2969 13-ethyl-11-methylene-18, 19-dinor-17-pregn-4-en-20-yn-17-ol - 3-keto-Org 2969 13-ethyl-17-hydroxy-11-methylene-18, 19-dinor-17-pregn-4-en-20-yn-3-one - ethinyl oestradiol 17-ethinyl-1,3,5(10)-oestratriene-3,17-diol - lynestrenol 19-nor-17-pregn-4-en-20-yn-17-ol - norgestrel 13-ethyl-17-hydroxy-18,19-dinor-17-pregn-4-en-20-yn-3-one     

Alkyl phloroglucinol derivatives from Syzygium levinei and their differentiation-inducing activity

Eleven compounds have been identified from the whole plants of Syzygium levinei (Myrtaceae) on the basis of spectroscopic analysis and chemical methods. Among these compounds, (Z)-1-(2,6-dihydroxy-4-methoxyphenyl)-oct-5-en-1-one (1), (3 E,7 Z)-1-(2,6-dihydroxy-4-methoxyphenyl)-deca-3,7-dien-1-one (2), and 1-(2-O-beta-D-glucopyranosyl-5,6-dihydroxy-4-methoxyphenyl)-octan-1-one (3) were characterized as new alkyl phloroglucinol derivatives, and 1-(2,6-dihydroxy-4-methoxyphenyl)-hexan-1-one (4) was identified as a new natural product. Compounds 1, 2, and 4 were found to inhibit the proliferation of leukemia K562 and HL-60 cells and also to induce the differentiation of K562 cells as determined by the appearance of matured morphology and the increase in the intracellular hemoglobin level.     

Three new diterpenoids from Coleus forskohlii Briq

Three new diterpenoids, forskolin G(2), forskolin H(3), forskolin I(4), were isolated from the whole plant of the Coleus forskohlii Briq., and their structures were elucidated as 1α,6β-diacetoxy-8,13-epoxylabd-14-en-11-one, 1α-hydroxy-6β,7β-diacetoxy-8,13-epoxylabd-14-en-11-one, and 1α,9α-dihydroxy-6β,7α-diacetoxy-8,13-epoxylabd-14-en-11-one on the basis of spectral data.     

Antioxidative 7-oxodehydropodocarpane-type trinorditerpenes from the bark of Taiwania cryptomerioides

Five new 7-oxopodocarpane-type trinorditerpenes together with 1beta,13,14-trihydroxy-8,11,13-podocarpatrien-7-one ( 1) were isolated from the bark of Taiwania cryptomerioides. By using NMR and other spectral methods, the structures of five new compounds, 14-hydroxy-13-methoxy-8,11,13-podocarpatriene-3,7-dione ( 2), 1beta,14-dihydroxy-13-methoxy-8,11,13-podocarpatrien-7-one ( 3), 13,14-dihydroxy-8,11,13-podocarpatriene-3,7-dione ( 4), 3beta,13,14-trihydroxy-8,11,13-podocarpatrien-7-one ( 5), and 1beta,13,14-trihydroxy-8,11,13-podocarpatriene-2,7-dione ( 6), were elucidated. Compounds 1, 4, 5, and 6 exhibited strong antioxidative activity.     

New beyerane and isopimarane diterpenoids from Rhizophora mucronata

Chemical examination of the ethyl acetate extract of the roots of Rhizophora mucronata collected from Mangalore Coast resulted in the isolation of three more new diterpenoids, rhizophorins C-E (1-3) in addition to the two, rhizophorin A, (6R,11S,13S)-6,11,13-trihydroxy-2,3-seco-14-labden-2,8-olid-3-oic acid, and rhizophorin B, ent-3beta,20-epoxy-3alpha,18-dihydroxy-15-beyerene, recently reported. The structures of the new diterpenoids have been elucidated by a study of their physical and spectral data as 17-hydroxybeyer-15-en-3-one (1), 3beta,17-diacetoxy-15-beyeren-2-one (2) and 3beta,6alpha-diacetoxy-8(14),15-isopimaradien-2-one (3).     

海绵共生真菌Penicillium janthinellum LZDX-32-1抗乙型肝炎病毒次生代谢产物研究

目的 探究Xest,ospongia testudinaria海绵共生真菌Penicillium janthinellum LZDX-32-1的次生代谢产物及其抗乙型肝炎病毒(HBV)活性。方法 对菌株进行发酵,运用现代色谱学方法(硅胶柱色谱、凝胶柱色谱、C-18反相柱色谱,以及半制备HPLC)对发酵产物进行分离,运用现代波谱学技术(核磁共振、质谱、紫外等),结合文献数据对比鉴定了化合物的结构;采用HepG2.2.15细胞模型的HBV DNA 水平用实时定量 PCR测定评价化合物的抗HBV活性。结果 分离鉴定了11个化合物,分别为：penialidin C (1)、penialidin A(2)、6-(2,4-dihydroxy-6-methylphenyl)-4-hydroxy-2-pyrone(3)、trans-3,4-dihydro-3,4,8-trihydroxynaphthalen-1(2H)-one(4)、epi-?isoshinanolone (5)、 pyrenocine B(6)、FK17-P2b1(7)、cordyol C(8)、diorcinol(9)、p-acetamidobenzoic acid(10)、2-(2-oxoindolin-3-yl)-acetamide(11)。 活性评价结果报道化合物5显示有抗HBV活性。结论 Xestospongia testudinaria海绵共生真菌 Penicillium janthinellum LZDX-32-1能够代谢产生具有抗HBV活性的次生代谢产物。     

Anti-platelet aggregation alkaloids and lignans from Hernandia nymphaeifolia

A new aporphine, N-(N-methylcarbamoyl)-O-methyl-bulbocapnine (1), together with seven known compounds, (-)-5'-methoxypodorhizol (2), a mixture of beta-sitosterone (3) and stigmasta-4,22-dien-3-one (4), a mixture of 3 beta-hydroxystigmast-5-en-7-one (5) and 3 beta-hydroxystigmasta-5,22-dien-7-one (6), and a mixture of 6 alpha-hydroxystigmast-4-en-3-one (7) and 6 alpha-hydroxystigmasta-4,22-dien-3-one (8), were isolated in continuing studies on the trunk bark of Formosan Hernandia nymphaeifolia. The structures of these compounds were determined through spectral analyses. In addition, the previously reported six alkaloids, laurotetanine, oxohernagine, thalicarpine, reticuline, (+)-vateamine-2'-beta-N-oxide, (+)-hernandaline and six lignans, (+)-epiaschantin, (+)-epimagnolin, (+)-epiyangambin, (-)-hernone, (-)-yatein, (-)-deoxypodophyllotoxin were demonstrated to have anti-platelet aggregation activity.