Uterine estrogen receptor binding of catecholestrogens and of estetrol (1,3,5(10)-estratriene-3,15alpha,16alpha,17beta-tetrol).

The binding affinities for the catecholestrogen metabolites of estradiol and of their methyl ethers for the rat uterine cytosol estrogen receptors were examined. Similarly the binding of the fetal estradiol metabolite, 15alpha-hydroxyestriol (estertrol) was also measured. All of the catecholestrogens showed binding affinities far in excess of their uterotrophic potency. This is different from estriol, the product of the alternative metabolic pathways and suggests that the direction of estradiol metabolism may have an important role in the modulation of estrogenic activity of the female sex hormone.     

The preparation of 2,4-dibromoestra-1,3,5(10),6-tetraene-3,17 beta-diol diacetate.

[2,4,6,7-3H4]Estradiol is used for the quantitative determination of estradiol receptors. The synthesis of 2,4-dibromoestra-1,3,5(10),6-tetraene-3,17 beta-diol 3,17-diacetate, the convenient precursor of [2,4,6,7-3H4]estradiol, is described. 6-Alkoxy compounds were also prepared and investigated.     

Mass spectrometry of ring D hydroxylated 3-methoxy-1,3,5(10)-estratrienes

Mass spectra and fragmentation patterns of the epimeric 17-, 16-, 15- and 14-hydroxy derivatives of 3-methoxy-1,3,5(10)-estratriene are compared. The main fragmentation pathways are differently influenced, depending on the position of the hydroxy group. The different configuration of the hydroxy groups is reflected only in the spectra of the epimeric 15- and 14-hydroxy compounds. Possibilities of mass spectrometric differentiation between the hydroxyestratrienes are discussed.     

Conjugate hydrocyanation of 17-acetyl-11-carbomethoxygona-1,3,5(10),13(17)-tetraenes

The conjugate hydrocyanation of 17-acetylgona-11-carbomethoxy-1,3,5(10),13(17)-tetraenes using diethylaluminum cyanide (Nagata reaction) is reported. This methodology has allowed the introduction of an angular cyano group at the C-13 position of the steroid skeleton. Subsequent reduction of the nitrile group yielded various functionalized steroids. One of them, 22 bears the natural trans/anti/trans stereochemistry and possesses an hydroxyl and aminomethyl functionalities in the positions 11beta and 13beta, respectively. The characteristic (1)H and (13)C NMR spectroscopic features of the synthesized steroids are reported.     

Cytotoxic steroids with antiestrogenic activity of the 11alpha-acyloxyestra-1,3,5(10)-triene series

Esterification of 3-hydroxyl group in 11-acyloxyestra-1,3,5(10)-trienes with p-[bis(2-chloroethyl)amino]phenylacetic acid led to antitumor steroids displaying antiestrogenic and cytotoxic activities. Our substances exhibit their activities on the model of murine mammary adenocarcinoma Ca-755, with inhibition of the tumor growth being 94-99%. A new approach was used for the 11alpha-hydroxylation of estra-1,3,5(10)-trienes.     

17Beta-hydroxy-11alpha-(3'-sulfanylpropyl)oxy-estra-1,3,5(10)- trien-3-yl sulfamate--a novel hapten structure: toward the development of a specific enzyme immunoassay (EIA) for estra-1,3,5(10)-triene-3-yl sulfamates.

The title compound 17 has been synthesized for the use as hapten in the development of a competitive enzyme immunoassay for estrogen sulfamates. The synthesis started from estradiol diacetate 2. Oxyfunctionalization at C-11 to give 11alpha-hydroxy steroid 8 was accomplished by hydroboration/alkaline hydrogen peroxide oxidation of the 9(11)-dehydro derivative 7, which was obtained from compound 2 via 9-hydroxylation with dimethyldioxirane. After transformation of compound 8 into the allyl ether 9, the side chain was thio-functionalized at the omega-position affording the thioate 11 in two steps. Selective silylether deprotection at position 3 followed by sulfamoylation gave the sulfamate 19, which in turn was demasked at position 17 and treated with sodium borohydride/aluminum chloride to liberate the side chain thiol. Alternatively, title compound 17 was synthesized via the disulfides 13-16. For the preparation of the immunogen the title compound 17 was coupled to bovine gamma globulin in a two-step procedure using an amine and thiol specific bifunctional crosslinker. The immunization of rabbits resulted in the formation of antibodies which clearly discriminated the sulfamoylated estrogens from the non-esterified estrogens. The use of a biotinylated hapten derivative as a tracer in combination with a streptavidin-peroxidase-tetramethylbenzidine based detection system allowed the measurement of estradiol 3-sulfamate (1) in the range of about 1 to 1000 pg/well.     

First total synthesis of (+/-)-3-aza-11-oxa-1,3,5(10)-trieno steroids

We set out to describe a new and versatile method for preparing 3-aza-11-oxa-1,3,5(10)-trieno steroids via an intramolecular Diels-Alder cycloaddition of o-quinodimethanes as the key step. The characteristic 1H and 13C NMR spectroscopic features of the synthesized compounds are reported.     

Preparation of 14,15-secoestra-1,3,5(10)-trien-15-ynes, inhibitors of estradiol dehydrogenase

The conversion of estrone to 14,15-secoestratrien-15-ynes, inactivators of estradiol dehydrogenase from human term placenta, is described. The optically pure precursor 7-acetoxy-octahydro-2-phenanthrenecarboxylic acid methyl ester is prepared from estrone in five steps and 40% yield. The unsubstituted propargylic secoestratriene diol, a mechanism-based inactivator of estradiol dehydrogenase, and the corresponding acetylenic ketone, an affinity label inactivator of the same enzyme, arise from the phenanthrene ester in three and four steps. The propargylic secoestratriene diol also competes with [3H]estradiol for binding to calf uterus estrogen receptor and possesses weak uterotrophic activity.     

Fungal interactions with the explosive RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine)

The bacterially mediated, anaerobic biodegradation of the explosive RDX (hexahydro 1,3,5 trinitro-1,3,5-triazine) is well established. Reports of successful mineralization of RDX by white rot fungi, and the enhanced transformation of RDX in stirred as compared to static composts, led us to study the possible aerobic role of several filamentous fungi in RDX biodegradation.Cladosporium resinae, Cunninghamella echinulata varelegans, Cyathus pallidus andPhanerochaete chrysosporium were grown in the presence of 50 and 100 g ml^–1 of RDX on a vegetable juice agar. Little inhibition of radial growth was observed, while control cultures with TNT exhibited substantial inhibition. When 100 g ml^–1 of RDX was added to pre-grown mycelia in a nonlignolytic liquid medium, between 12 and 31% was lost after 3 days. In similar experiments using¹⁴C-RDX, most of the label remained in the organic fraction, and little or none was found in the aqueous fraction, the volatile fraction or incorporated into cell walls. Although disappearance of RDX was observed for all four species tested, there was no evidence of mineralization. Mixed cultures of microorganisms, including both bacteria and fungi, merit further study as agents for the decontamination of munitions-contaminated soils.     

Efficient synthesis of 17-acetyl-13-(p-bromophenyl)-3-methoxy-11,11-bis(methoxycarbonyl)gona-1,3,5(10)-trienes

We described an efficient synthesis of (8β,9β,14β)-17β-acetyl-13β-p-bromophenyl-11,11-di(methoxycarbonyl)-3-methoxygona-1,3,5(10)-triene, (8β,9α,14β)-17β-acetyl-13β-p-bromophenyl-11,11-di(methoxycarbonyl)-3-methoxygona-1,3,5(10)-triene, (8β,9β,14β)-13 β-p-bromophenyl-11,11-di(methoxycarbonyl)-17β-(2-hydroxyethyl)-3-methoxygona-1,3,5(10)-triene, and (8β,9β,14β)-13β-p-bromophenyl-11,11-di(methoxycarbonyl)-17β-(2-oxoxyethyl)-3-methoxygona-1,3,5(10)-triene in five or six steps from 1-iodo-4-methoxybenzocyclobutene and readily available materials.     

Dynamic motions of 1,6-diphenyl-1,3,5-hexatriene in interdigitated C(18):C(10)phosphatidylcholine bilayers.

This study investigates the dynamic behavior of 1,6-diphenyl-1,3,5-hexatriene (DPH) in C(18):C(10)phosphatidylcholine [C(18):C(10)PC] bilayers. C(18):C(10)PC is an asymmetric mixed-chain phosphatidylcholine known to form mixed-interdigitated structures below the transition temperature and form partially interdigitated bilayers above the transition temperature. The rotation of DPH in C(18):C(10)PC has been described in terms of the thermal coefficient of rotation using the modified Y-plot method which takes into account the limiting anisotropy value. During the phase transition of C(18):C(10)PC, DPH exhibits a thermal coefficient b2M = 0.41 - 0.51 degrees C-1 which is similar to the b2M values obtained with noninterdigitated phosphatidylcholine bilayers. Differential polarized phase-modulation fluorometry has also been employed to study the dynamic behavior of DPH in C(18):C(10)PC in real time. The data show that DPH contains considerable motion in the highly ordered mixed interdigitated bilayers. The DPH motion steadily increases with an increase in temperature as shown by the rotational correlation time, and the wobbling diffusion constant. However, the limiting anisotropy, the order parameter, and the width of the lifetime distribution undergo an abrupt decrease, and a corresponding abrupt increase in the cone angle, at approximately 16 degrees C. This temperature range is near the onset temperature of the phase transition as determined by differential scanning calorimetry. The rotational parameters show strong hysteresis on heating and cooling. All the rotational parameters derived from DPH fluorescence in mixed interdigitated C(18):C(10)PC exhibit magnitudes similar to those obtained from noninterdigitated gel phases of symmetric diacylphosphatidylcholines.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synthesis of 3,6α,16α-trihydroxy-1,3,5(10)-estratrien-17-one 6-hemisuccinate and [6,73H]-3,16α-dihydroxy-1,3,5(10)-estratrien-17-one

The preparation of high specific activity ³H labeled 16α-hydroxyestrone and of the 6-hemisuccinate of the ketol metabolite of estradiol is described. The synthetic procedures which were required to produce these essential ingredients of a radioimmunoassay for 16α-hydroxyestrone required multistep syntheses because of the multifunctional and labile nature of the metabolite.