菌物中麦角甾类化合物的研究进展

甾类化合物广泛存在于自然界中，是一类很重要的天然产物。在菌物中，甾类化合物不仅种类多样，含量也相当丰富，尤其是由28个碳原子组成的麦角甾类化合物更具有代表性，是菌物的特征性成分。文中对国内外文献报道的菌物中麦角甾类化合物的种类进行了归纳总结。     

Synthesis and biological properties of D-homo-6-oxa-8α-analogues of steroid estrogens

Modifications of D-homo-6-oxa-8α-analogues of steroid estrogens were found to lead to a complete loss of the uterotropic and hypertriglyceridemic activities. These compounds may be promising for the design on their basis of inhibitors of the steroid hormone metabolism and transporters of other compounds to the estrogen target organs.     

Increasing the amphiphilicity of an estradiol based steroid structure by Barbier-allylation--ring-closing metathesis--dihydroxylation sequence

Polyhydroxylated steroids, such as brassinosteroids, phytoecdysteroids and steroid saponins, are structurally attractive compounds possessing a number of interesting biological properties. Accordingly, development of synthetic procedures to build steroid based structures mimicking the naturally occurring hydrophilic steroids is of topical interest. In the present work, a D-secoestrone derivative was modified further by Barbier-allylation - ring-closing metathesis - dihydroxylation sequence with the aim to prepare steroid based structures with limited hydrophilicity. A straightforward synthesis route was developed with the isolated yield for each step ranging from good to excellent. All compounds prepared were fully characterized by NMR spectroscopic techniques and completely assigned (1)H and (13)C spectra are reported herein. Finally, the effects of the synthesized amphiphilic steroid derivatives on the proliferation of cancer cells are reported and discussed.     

Ent-16α-methylamino-3-methoxy-1,3,5(10)-estratrien-17β-ol hydrochloride, a selective anti-arrhytmic enantiosteroid

The title compound and its antipode with natural steroid configuration were synthesized and tested. Both compounds showed equal potency as anti-arrhythmic compounds. An additional effect found for the steroid with natural configuration was its plasma cholesterol lowering activity in the rat. This suggests that enantiosteroids may have a more selective action than the steroids with natural configuration     

Development of adenosine responsiveness after isolation of Leydig cells

Effects of adenosine and related compounds on the regulation of steroid production by isolated Leydig cells have been investigated. Steroid production by freshly isolated Leydig cells from testes of immature or mature rats and mice, or from Leydig tumor tissue could not be stimulated with adenosine, nicotinamide-adenine dinucleotide (phosphate) [NADPH, NAD(P)] or N6-(1-2-phenylisopropyl)-adenosine (PIA) (50 microM), whereas luteinizing hormone (LH) stimulated steroid production more than 10-fold. After 24 h incubation all adenosine-related compounds, but not inosine, stimulated steroid production to 20-100% of the maximal LH-stimulated activity. LH- or 22R -hydroxycholesterol-stimulated steroidogenesis in Leydig cells from immature rats did not decrease during the 24-h culture period, whereas ATP levels increased. The first significant effect of adenosine on steroid production in these cells was found after an incubation period of 3 h. In cells incubated for 1 h and 24 h, LH stimulated cyclic adenosine 3':5'-monophosphoric acid (cAMP) production 10-fold. Significant effects of adenosine and PIA on cAMP production or protein phosphorylation could only be shown in cells incubated for 24 h. Effects of adenosine on Leydig cells in intact testis tissue of immature rats could not be determined. The results suggest that after isolation of Leydig cells, specific alterations in the cell membrane occur, causing increased sensitivity to adenosine and related compounds. Adenosine apparently does not play a role in the role of steroid production in Leydig cells in vivo.     

Igor' Vladimirovich Torgov. Corresponding member of the Russian Academy of Sciences. On his 95th birthday

The late 1940s and the early 1950s were marked by an extraordinary rise in the interest of scientists worldwide (chemists, biologists, and physicians) in steroid hormones, one of the types of natural compounds later referred to as low-molecular bioregulators. The steroid compounds constructed on the basis of the cyclopentanoperhydrophenanthrene skeleton turned out to be the most universal of all known representatives of natural compounds, exhibiting a high regulatory activity in the broadest set of physiological processes. At the very beginning of study of this class of compounds, it was found that they are responsible for sexual development and maturation, reproduction, water-salt balance, and exchange of carbohydrates. The spectrum of their activities was extended ever further, and, nowadays, perhaps no known vital function of living organisms remains in which, in some way, steroids are not involved.     

Binding of steroid hormones and anabolic agents to bovine sex-hormone binding globulin

Binding of some selected steroids and anabolic agents to bovine sex-hormone binding globulin (SHBG) was investigated. SHBG binding affinities, relative to the reference hormone 5 alpha-dihydrotestosterone, were estimated for the compounds. The results demonstrate that binding of steroid hormones to SHBG is facilitated by the 17 beta-hydroxyl group, possibly involving hydrogen binding, and by the methyl group at C-19 of the steroid moiety. Structural modifications at C-17 of a steroid molecule involving esterification, epimerization or reduction of the 17 beta-hydroxyl group, or introduction of a bulky 17 alpha group have the effect of decreasing the SHBG binding affinity of the steroid molecule.     

Identification of small-molecule inhibitors of the steroidogenic acute regulatory protein (STARD1) by structure-based design

A homology model of the steroidogenic acute regulatory protein (STAR)-related lipid transfer (START) domain of STARD1 was built, and the cholesterol binding site was identified. Structure-based design studies were performed to identify small molecule inhibitors of the START domain. The lead compounds were selected based on cAMP-induced, but not 22R-hydroxycholesterol-supported, inhibition of steroid synthesis by 50% at 10 μM. The results obtained by molecular docking & dynamics show a good correlation between bioactivity, docking scores and calculated binding energies of ligand-protein complexes. The best active compounds will be optimized further and used to develop potential drugs to control excessive steroid formation.     

Anabolic utilization of steroid hormones in Helicobacter pylori

In this study, we have demonstrated that Helicobacter pylori absorbs a steroid prehormone (pregnenolone) and two androgens (dehydroepiandrosterone and epiandrosterone), glucosylates these steroids, and utilizes glucosyl-steroid hormone compounds as the membrane lipid components. The only common structure among the steroid prehormone and the two androgens is a 3β-OH in the steroid framework. Our results indicate that the 3β-OH in the steroid hormones is a crucial conformation required for steroid glucosylation by H. pylori . In addition, we found that H. pylori absorbs and holds estrogens possessing 3-OH (estrone and estradiol) into the membrane. The effective absorption of estrogen into the membrane appeared to be controlled by the number of hydroxyl groups modifying the steroid framework. In contrast, H. pylori induced neither membrane absorption nor glucosylation of the other steroid hormones possessing 3=O (progesterone, androstenedione and testosterone) or 3α-OH (androsterone). These results indicate that H. pylori selectively absorbs 3β-OH and 3-OH steroid hormones, and utilizes only 3β-OH steroid hormones as the materials for glucosylation.     

Method for combined analysis of profiles of conjugated progesterone metabolites and bile acids in serum and urine of pregnant women

A method for analysis of profiles of conjugated progesterone metabolites and bile acids in 10 ml of urine and 1–4 ml of serum from pregnant women is described. Total bile acids and neutral steroids from serum and urine were extracted with octadecylsilane-bonded silica. Groups of conjugates were separated on the lipophilic ion-exchanger triethylaminohydroxypropyl Sephadex LH-20 (TEAP-LH-20). Fractions were divided for steroid or bile acid analyses. Sequences of hydrolysis/ solvolysis and separations on TEAP-LH-20 permitted separate analyses of steroid glucuronides, monosulfates and disulfates and bile acid aminoacyl amidates, sulfates, glucuronides and sulfate-glucuronides. Radiolabelled compounds were added at different steps to monitor recoveries and completeness of separation, and hydrolysis/solvolysis of conjugates was monitored by fast-atom bombardment mass spectrometry. The extraction and solvolysis of steroid disulfates in urine were studied in detail, and extraction recoveries were found to be pH-dependent. Following methylation of bile acids, all compounds were analysed by capillary gas chromatography and gas chromatography—mass spectrometry of their trimethylsilyl ether derivatives. Semiquantification of individual compounds in each profile by gas—liquid chromatography had a coefficient of variation of less than 30%. The total analysis required 3 days for serum and 4 days for urine.     

Synthesis and anti-HIV activity of cosalane analogues incorporating two dichlorodisalicylmethane pharmacophore fragments

A new series of cosalane analogues incorporating two fragments of the dichlorodisalicylmethane pharmacophore has been synthesized. In order to identify the position for the attachment of the pharmacophore fragments to the steroid ring that results in the most potent analogues, two types of compounds were designed. In the first type, the two pharmacophore fragments were attached at C-3 and C-17 of the steroid ring by using appropriate linker units. In the second type, both pharmacophore groups were connected to C-3 of the steroid through an alkenyl chain containing an amide moiety. All of the new compounds displayed antiviral activity versus HIV-1(RF), HIV-1(IIIB), and HIV-2(ROD) in cell culture. The relative potencies of the compounds resulting from the two attachment strategies were found to depend on the viral strain as well as the cell type. Overall, the attachment of the second pharmacophore did not result in either a large gain or a large loss in anti-HIV activity, and the results are therefore consistent with the hypothesis that the two pharmacophores act independently, and one at a time, with positively charged amino acid side chains present on the surface of gp120 and CD4.     

甾类化合物微生物转化与分解代谢机制研究进展

甾类化合物具有重要的生理医药作用,市场需求巨大。甾类化合物及其关键甾类药物通过微生物转化制备工艺较化学合成法具有区域立体选择性、减少合成步骤、缩短生产周期、提高收率以及生态友好等优点逐步被应用,然而甾类物质微生物分解代谢机制还有待进一步深入探索研究并确定。本文从甾类化合物结构种类与主要来源、生理功能、微生物转化与分解代谢机制的研究等方面进行了归纳,着重解析甾类化合物分解代谢过程关键酶系及其分子作用机制,为甾药化合物生产菌种改造与工程菌构建,以及微生物转化工业化生产工艺的开发提供参考。     

Synthesis, structure, and biological properties of some 8alpha-analogues of steroid estrogens with fluorine in position 2

A total synthesis of 8alpha analogues of steroid estrogens with fluorine in position 2 was achieved. Structural features of these compounds were studied by the example of 17beta-acetoxy-2-fluoro-3-methoxy-8alpha-estra-1,3,5(10)-triene. It was shown that the 8alpha analogues of 2-fluorosubstituted steroid estrogens have a low uterotropic activity and retain the osteoprotective and hypocholesterolemic activities.     

New fluorogenic substrate for the first continuous steroid sulfatase assay

The screening for new inhibitors of steroid sulfatase requires an efficient test system. To overcome the shortcomings of the available discontinuous fluorimetric assay, several coumarin-type compounds were investigated as potential new substrates. 3,4-Benzocoumarin 7-O-sulfate was found to have appropriate substrate properties for the establishment of the first direct continuous assay of steroid sulfatase.     

Synthesis,structure, and biological properties of some 8α analogues of steroid estrogens with fluorine in position 2

A total synthesis of 8α analogues of steroid estrogens with fluorine in position 2 was achieved. Structural features of these compounds were studied by the example of 17β-acetoxy-2-fluoro-3-methoxy-8α-estra-1,3,5(10)-triene. It was shown that the 8α analogues of 2-fluorosubstituted steroid estrogens have a low uterotropic activity and retain the osteoprotective and hypocholesterolemic activities.     

A new class of long-acting hormonal steroid preparation: synthesis of dimeric ethynodiol and nortestosterone, of dimeric and trimeric androgens and of some dimeric combinations of steroids

Dimeric steroid esters containing an estradiol molecule coupled with a molecule of ethynodiol, cortexo lone, or ethinylandrostenediol respectively, were synthesized. The same method was also used for dimerization of ethynodiol, nortestosterone, and testosterone, and for the preparation of a trimeric androgen consisting of two molecules testosterone linked by one molecule 5-androstene-3β, 17β-diol. The method is based upon the direct esteriflcation of steroid alcohols with steroid hemisuccinates in the presence of N,N'-carbonyldiimidazole. The results of preliminary experiments concerning the biological activity of some of these compounds showed a considerable prolongation of hormonal effect as compared to the respective monomer.     

Cytotoxicity and gene expression profiles of novel synthesized steroid derivatives as chemotherapeutic anti-breast cancer agents

Anti-cancer agents which combine two biologically active compounds in one such as steroidal heterocyclic derivatives attain both hormone and cytotoxic effects on cancer cells. The aim of the present study is to synthesize and evaluate new potential chemotherapeutic anti-breast cancer agents. Several pyridazino-, pyrimido-, quinazolo-, oxirano- and thiazolo-steroid derivatives were synthesized. The structure of the novel steroid derivatives was confirmed using the analytical and spectral data. The most structurally promising of the novel synthesized steroid derivatives, compounds 8, 12, 17, 20, 22c, 24c, 30a and 30b, were investigated individually as anti-breast cancer agents against human breast cancer cells (MCF-7) using sulforhodamine B (SRB) assay. The tested compounds 17, 20, 22c and 8 showed potent broad spectrum cytotoxic activity in vitro after 48 h incubation. Compound 17 (IC(50)=2.5 μM) exhibited more inhibitory influence on MCF-7 growth than the reference drug doxorubicin (Dox) (IC(50)=4.5 μM) after 48 h incubation. Also, the present study showed that all the tested steroid derivatives exhibited significant depletion with various intensities in gene expression of breast cancer related genes (VEGF, CYP19 and hAP-2γ). Noteworthy, compounds 17, 20 and 22c showed the most pronounced effect in this respect.     

Progesterone analogs influence germination of Clostridium sordellii and Clostridium difficile spores in vitro

Clostridium sordellii and Clostridium difficile are closely related anaerobic Gram-positive, spore-forming human pathogens. C. sordellii and C. difficile form spores that are believed to be the infectious form of these bacteria. These spores return to toxin-producing vegetative cells upon binding to small molecule germinants. The endogenous compounds that regulate clostridial spore germination are not fully understood. While C. sordellii spores require three structurally distinct amino acids to germinate, the occurrence of postpregnancy C. sordellii infections suggests that steroidal sex hormones might regulate its capacity to germinate. On the other hand, C. difficile spores require taurocholate (a bile salt) and glycine (an amino acid) to germinate. Bile salts and steroid hormones are biosynthesized from cholesterol, suggesting that the common sterane structure can affect the germination of both C. sordellii and C. difficile spores. Therefore, we tested the effect of sterane compounds on C. sordellii and C. difficile spore germination. Our results show that both steroid hormones and bile salts are able to increase C. sordellii spore germination rates. In contrast, a subset of steroid hormones acted as competitive inhibitors of C. difficile spore germination. Thus, even though C. sordellii and C. difficile are phylogenetically related, the two species' spores respond differently to steroidal compounds.     

Enzymatic breakdown of steroids during 11 alpha- and 11 beta-hydroxylation of the 21-acetate of Reichstein's "substance S" by Tieghemella orchidis (fam. Mucoraceae)]

By-products of the transformation were studied, which were formed in the process of 11alpha and 11beta-hydroxylation of 21-acetate of the Reichstein substance "S" by the culture of Tieghemella orchidis. The following steroid derivatives, known before, were isolated: epihydrocortisone, 6beta-hydroxy derivative of the Reichstein substance "S", hydrocortisone, cortisone. The following steroid derivatives were isolated from the initial chloroform extract of the fermentation medium, and identified: prednisolone, androstendione, testololactone, and 1,2-dehydro derivatives of the Reichstein substance S" and its 21-acetate. The quantitative ratio between the products of transformation depended on the conditions of growth and transformation. These compounds seem to be intermediate metabolites in the course of the steroid destruction via the pathway common for fungi and involving the destruction of D ring of the steroid molecule.     

目前处于临床研究的抗肿瘤活性海洋天然产物

人们从海洋生物中分离出数万种包括萜类、甾类、多肽、聚醚类、大环内酯类、生物碱类等具有活性的化合物,目前有近二十种抗肿瘤活性化合物正处于临床研究。