Biliary Excretion of 17β-Estradiol 17β-d-Glucuronide Is Predominantly Mediated by cMOAT/MRP2

Purpose. The mechanism for the biliary excretion of 17-estradiol170-d-glucuronide (E₂17G), a cholestatic metabolite of estradiol, isstill controversial. The purpose of the present study is to examine thetransport of E₂17G across the bile canalicular membrane. Methods. We examined the uptake of [³H]E₂17G by isolatedcanalicular membrane vesicles (CMVs) prepared from Sprague-Dawley (SD)rats and Eisai Hyperbilirubinemic rats (EHBR) whose canalicularmultispecific organic anion transporter/multidrug resistance associatedprotein 2 (cMOAT/MRP2) function is hereditarily defective. Also,in vivo biliary excretion of intravenously administered [³H]E₂17Gwas examined. Results. In CMVs prepared from SD rats, but not from EHBR, amarked ATP-dependent uptake of [³H]E₂17G was observed.Moreover, E₂17G competitively inhibited the ATP-dependent uptake of[³H]2,4-dinitrophenyl-S-glutathione (DNP-SG). In addition, nosignificant inhibitory effect of verapamil (100 M) and PSC-833 (5 M) onthe uptake of [³H]E₂17G was observed. In vivo, the biliary excretionof intravenously administered [³H]E₂17G was severely impaired inEHBR while the biliary excretion of [³H]E₂17G in SD rats wasreduced by administering a cholestatic dose (10 mol/kg) unlabeledE₂17G, but not by PSC-833 (3 mg/kg). Conclusions. The transport of E₂17G across the bile canalicularmembrane is predominantly mediated by cMOAT/MRP2.     

β受体反应亢进症17例报告

乙型肾上腺能受体(β受体)反应亢进症(简称β亢症)临床上较为常见.并常与甲亢、心肌炎、冠心病等混淆。我们于1984年3月～1986年10月共治疗该病17例,现报道如下。一、临床资料 20～50岁,其中20～40岁14例;女14例,男3例;病程1～5年,临床主要表现为神经及循环系统症状,17例均有头晕、心悸、胸闷,失眠13例,神经质11例,胸部刺痛9例,乏力14例,     

17-β-雌二醇治疗更年期综合征

目的 :评估 17 β 雌二醇治疗更年期综合征的疗效。方法 :84例患有更年期综合征的妇女分 2组 ,试验组 33例 ,年龄 4 8a±s5a ,服用 17 β 雌二醇1mg ,po ,qd ,共 3个周期 ,对照组 5 1例 ,年龄 4 8a±5a ,服用共轭雌激素 0 .62 5mg ,ро ,qd ,共 3个周期 ,2组每周期d 15均加用醋酸甲羟孕酮 4mg ,ро ,qd× 14d ,更年期综合征以改良Kupperman评分评估 ,治疗前后检查血雌二醇、促卵泡激素。结果 :治疗 3个周期后 ,2组Kupperman评分分别下降87% ,80 % ,雌二醇上升至卵泡中期水平 ,促卵泡激素显著下降 ,2组比较差异无显著意义 (P >0 .0 5 )。结论 :17 β 雌二醇治疗更年期综合征与共轭雌激素疗效相似。     

高浓度17β-雌二醇诱导巨噬细胞凋亡

目的　考察高浓度 17β 雌二醇 (E2 )对巨噬细胞的影响。方法　用Hoechst332 5 8进行染色质荧光染色 ;扫描电子显微镜观察细胞形态 ;激光共聚焦显微镜检测胞质内钙离子浓度 [Ca2 + ]i 及线粒体膜电位。结果　荧光染色及扫描电镜显示E2 (≥ 1μmol·L-1)作用后巨噬细胞出现多种凋亡表现。E2 1μmol·L-1作用后 ,细胞 [Ca2 + ]i 明显升高 ,且有剧烈的波动。E2 1μmol·L-1使线粒体膜电位下降。结论　高浓度的E2 (≥ 1μmol·L-1)能诱导巨噬细胞的凋亡 ,并且与[Ca2 + ]i 的变化及线粒体膜电位的下降有关     

Stability of Extruded 17β-Estradiol Solid Dispersions

Recrystallization is one of the main problems concerning the stability of solid dispersions. Different analytical methods were applied showing that no recrystallization occurred after treating melt extruded solid dispersions with 17β-Estradiol as the model drug with heat or water vapor. A skillful choice of excipients—a combination of polymers and additives—could be the reason for improving the stability. The requirements of the USP 23 for Estradiol tablets of 75% dissolved drug after 60 min were fulfilled after storing the tablets for 6 months at 40°C/75% RH. By observing the change in glass transition temperature, DSC analysis showed that the solid dispersions were stable against thermal stress. Isothermal microcalorimetry as well as moisture absorption gravimetry were methods to prove the stability of the solid dispersions against water vapor.     

Gender differences in response to chronic treatment with 17β-oestradiol and 17β-aminoestrogen pentolame on hemostasis in rats

Objectives:

This work evaluated chronic treatment with 17β-oestradiol (E₂) and 17β-aminoestrogen pentolame (AEP) on prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), and fibrinogen concentration (FIB). Male (M) and ovariectomized (Ovx) Wistar rats were used to explore gender differences in the pharmacological response.

Materials and Methods:

Rats (n = 12-18) were treated every third day during three months with E₂ (1, 10, 100 μg/kg), AEP (1, 10, 100, 500 μg/kg) or vehicle (propylenglycol 1 ml/ kg). PT, aPTT, TT, and FIB were measured using standardized techniques.

Results:

Chronic treatment with E₂ in male rats increased PT (4-7%; P < 0.05), decreased aPTT (9%; 100 μg/kg; P < 0.05) and decreased TT (5% at 100 μg/Kg; P < 0.05). Chronic treatment with E₂ in ovariectomized female rats decreased PT (3-4%; P < 0.05), did not induce significant changes on aPTT and decreased TT in a dose dependent manner (12-27%; P < 0.05). Chronic treatment with AEP in male rats did not alter PT, increased aPTT in a dose dependent manner (5-16%; P < 0.05), and decreased TT (5%; 500 μg/Kg; P < 0.05) while in female ovariectomized rats it decreased PT (5-9%; P < 0.05), increased aPTT (8-13%; P < 0.05) and decreased TT (6-13%; P < 0.05). E₂ and AEP decreased FIB in M and Ovx animals. Decreases in FIB by E₂ were more pronounced in male (15-18% P < 0.05) than in ovariectomized rats (10-14% P < 0.05). E₂ showed more potency than AEP, lowering FIB at 1 and 10 μg/kg doses. Both estrogens decreased FIB in ovariectomized animals (E₂, 10-14%, P < 0.05; AEP, 9% P < 0.05) and were reverted by increasing dosage.

Conclusions:

Gender influenced response to chronic treatment with E₂ and AEP on hemostatic parameters. PT and aPTT were the most affected parameters, demonstrating non-equivalence in the pharmacological response of M and Ovx rats.KEY WORDS: 17β-aminoestrogens, gender, hemostasis, oestradiol, rat     

Pharmacological profile of a 17β-heteroaryl-substituted neuroactive steroid

Rationale

In order to improve upon the pharmacological properties of the neuroactive steroid ganaxolone, it was used as the starting point in the design of novel neurosteroids that replace the 17β-acetyl side chain with an isoxazole bioisostere.

Objectives

UCI-50027 (3-[3α-hydroxy-3β-methyl-5α-androstan-17β-yl]-5-(hydroxymethyl)isoxazole) was designed as an orally active neuroactive steroid specifically targeted at the gamma-aminobutyric acid(A) receptor (GABA_AR).

Methods

UCI-50027 was tested in vitro in Xenopus oocytes expressing human GABA_ARs and in vivo as an anticonvulsant, for ataxic effects and for anxiolytic activity.

Results

In vitro, UCI-50027 dose-dependently enhanced the activity of GABA at human α₁β₂γ_2L, α₂β₁γ_2L, and α₄β₃δ GABA_ARs. Consistent with its action as a positive allosteric modulator (PAM), it had no direct activity in the absence of GABA. UCI-50027 protected against acute pentylenetetrazol (PTZ)-induced convulsions with an ED₅₀ of 6 mg/kg p.o. In the rotarod (RR) paradigm in mice, the AD₅₀ (the ataxic dose where half of the animals fail the RR test) was found to be 38 mg/kg p.o., giving a therapeutic index (TI = RR AD₅₀/PTZ ED₅₀)～6 versus 2.8 for ganaxolone. In the mouse-elevated plus maze (EPM) model for anxiety, UCI-50027 showed a minimum effective dose (MED) ≤0.3 mg/kg p.o. Thus, the TI (TI = RR AD₅₀/EPM MED) for the compound as an anxiolytic is ≥127 versus 3.3 for ganaxolone.

Conclusions

UCI-50027 is an orally active neuroactive steroid with pharmacological activity consistent with a GABA_AR PAM that has an improved separation between anticonvulsant/anxiolytic and rotarod effects, potent activity as an anticonvulsant and anxiolytic when compared to ganaxolone.     

17β-Hydroxysteroid dehydrogenase inhibitors: a patent review

Importance of the field: 17β-Hydroxysteroid dehydrogenases (17β-HSDs) mainly catalyze the reduction of C17-ketosteroids to their corresponding hydroxylated forms as well as the reverse reaction (oxidation). Able to convert inactive or less active steroid hormones into more potent ones and vice versa, certain 17β-HSDs play a key role, especially in the regulation of estrogen and androgen levels. The therapeutic potential of this enzyme family, especially for the treatment of breast cancer, prostate cancer, acne and osteoporosis, then stimulated the development of inhibitors of 17β-HSDs and important progress was achieved over the last years.

Areas covered in this review: This review article reports all patent applications related to the inhibitors of 17β-HSDs, including some articles needed to complement the information presented.

What the reader will gain: Readers will be informed about the role and function of 17β-HSDs in the first section and about the history of inhibitor development in the second section. Furthermore, in the third and main section, the readers will learn about the structures of patented inhibitors originating from different companies and academic groups.

Take home message: The increase in the number of 17β-HSD inhibitors reported in the last years augurs well for the future. The challenge is now to translate these results into clinical studies to allow determination of the therapeutic usefulness of 17β-HSD inhibitors.     

17β-estradiol decreases methylmercury-induced neurotoxicity in male mice

There is increasing evidence that health effects of toxic metals, including methylmercury (MeHg), differ in prevalence or are manifested differently in men and women. The present study was aimed at investigating the potential differential susceptibility of male and female Swiss mice against MeHg-induced neurotoxicity, which was evaluated by biochemical (cerebellar oxidative stress-related parameters) and behavioral (locomotor activity and motor performance) variables. We also aimed to evaluate the potential protective effects of 17β-estradiol against such toxicity in MeHg-exposed male animals. MeHg exposure (40mg/L, diluted in tap water, during 2 weeks) decreased locomotor activity and motor performance in both male and female animals, but such phenomena were higher in males. 17β-estradiol co-treatment (10μg/animal, in alternate days) prevented MeHg-induced locomotor deficits in males. MeHg exposure caused a significant increase (60%) in cerebellar lipid peroxidation in male mice, but did not in females. In close agreement, MeHg exposure decreased (43%) cerebellar glutathione peroxidase activity in males, but did not in females. These events were prevented by 17β-estradiol administration. Cerebellar GR activity was increased (25%) in MeHg-exposed males and such event was partially prevented by 17β-estradiol administration. These results indicate that the low susceptibility of female mice to the neurotoxicity elicited by MeHg is linked to neuroprotective effects of sex steroids, which appear to modulate the activities of glutathione-related enzymes. Our experimental observation corroborates previous epidemiological studies showing the greater developmental effects in male than in female humans exposed to MeHg.     

Nasal Absorption Enhancement of 17β-Estradiol by Dimethyl-β-Cyclodextrin in Rabbits and Rats

A new formulation for nasal administration containing 17-estradiol (E₂) with dimethyl--cyclodextrin (DMC) as a solubilizer and absorption enhancer is described. Nasal administration of this E₂-DMC formulation gave a significantly higher E₂ absorption than an E₂ suspension in both rabbits and rats. Relative to an intravenous injection of the E₂-DMC formulation, absolute bioavailabilities of 94.6 and 67.2% were calculated for the nasal E₂-DMC formulation in rabbits and rats, respectively. Differences in bioavailability may have resulted from differences in experimental animal conditions. The effects on human nasal ciliary activity of the E₂-DMC formulation were studied with an in vitro method. The formulation was found to exert only a minor effect on ciliary beat frequency. Thus, nasal delivery of E₂, using a cyclodextrin inclusion formulation, may have potential for clinical application, e.g., in the therapy of postmenopausal disorders.     

Eine Oestradiol-17β-Dehydrogenase in den Erythrocyten der Ratte

Zusammenfassung In Fortsetzung einer Untersuchungsreihe über Steroidfermente wird in der vorliegenden Arbeit über eine Oestradiol-17-Dehydrogenase in den roten Blutzellen der Ratte berichtet. Intakte Zellen reduzieren Oestron zu Oestradiol-17 und dehydrogenieren Oestradiol-17, dagegen nicht das 17-Epimere, zu Oestron. Zwischen den beidem Reaktionen stellt sich ein Gleichgewicht her, das unter physiologischen Bedingungen auf seiten des Reduktionsprodukts liegt. Die unter wechselnden Bedingungen angetroffene Gleichgewichtseinstellung wird vor allem von dem Verhältnis TPNH:TPN bestimmt. Dieses Verhältnis wiederum hängt vorwiegend von der Aktivität der Glucose-6-phosphat-Dehydrogenase ab. — Nebenreaktionen, insbesondere die Bildung von Oestradiol-17, lassen sich nicht nachweisen. Zwischen dem Blut von Ratten verschiedenen Geschlechts sind keine Aktivitätsdifferenzen. Die Erythrocyten von Ratten mit fortgeschrittenem Hepatom oder Yoshida-Sarkom enthalten weniger Ferment. praktisch der gesamte Gehalt des Bluts an Oestradiol-17-Dehydrogenase fällt auf die roten Blutzellen. Darstellung und Eigenschaften eines rohen Fermentpräparats aus Erythrocyten werden beschrieben. Die Ergebnisse werden unter den Gesichtspunkten Gleichgewichts-Einstellung mit Erythrocyten sowie Stereo- und Pyridin-nucleotid-Spezifität des Ferments diskutiert.Mit 5 TextabbildungenEin Teil der Ergebnisse wurde auf der Tagung der Gesellschaft für Physiologische Chemie am 24. September 1959 in Berlin vorgetragen.Meinem verehrten Lehrer, Herrn Professor P. Wels, in Dankbarkeit zum 70. Geburtstag gewidmet.     

长效避孕药——17β-甾醇酯的合成

本文报道了为寻找长效避孕药而设计的炔诺酮酯和18-甲基炔诺酮酯的合成方法。以双环己基碳二亚胺(DCC)为脱水剂,4-二甲氨基吡啶(DMAP)为接触剂,酸与醇直接反应,合成了五个17β-酯,收率为60～90%。     

17β雌二醇对氯胺酮所致神经元凋亡的影响

目的：研究17β雌二醇对氯胺酮所致原代培养皮质神经元凋亡的影响及其机制。方法原代培养皮质神经元,体外培养7 d,随机分为空白对照组（给予相同体积的DMSO）,雌二醇组（17β雌二醇终浓度为0.1μmol·L-1）,氯胺酮组（氯胺酮终浓度为100μmol·L-1）,氯胺酮＋雌二醇组（氯胺酮,17β雌二醇终浓度分别为100μmol·L-1,0.1μmol·L-1）。噻唑蓝（ MTT）法检测神经元存活率,Hoechest33258染色法检测皮质神经元凋亡,Western-blot 法测定cleaved-Caspase-3及Bcl-2蛋白表达。结果氯胺酮组神经元存活率（54.02±7.78）%,明显低于空白对照组;氯胺酮＋雌二醇组神经元存活率（88.09±6.54）%,明显高于氯胺酮组。神经元经Hoechest33258染色在荧光显微镜下观察,氯胺酮组神经元凋亡[凋亡率（49.50±4.34）%]较空白对照组明显增加,氯胺酮＋雌二醇组[凋亡率（15.74±3.40）%]较氯胺酮组神经元凋亡下降。氯胺酮组 cleaved-Caspase-3表达明显增加,Bcl-2明显下降,而氯胺酮＋雌二醇组较氯胺酮组cleaved-Caspase-3表达明显下降,Bcl-2表达明显升高。结论17β雌二醇通过抑制神经元凋亡对抗氯胺酮诱导的皮质神经元损伤,产生保护作用。     

17β-雌二醇预防绝经后妇女的骨质疏松症

本文旨在评价经皮肤给予三种剂量17β-雌二醇对预防绝经后妇女骨质疏松症的效果。127例绝经后妇女，年龄＞40岁。平素无骨骼疾病。随机接受17公雌二醇（0．025、0．05或0．ling／天）或安慰剂。全组中共有路例患者完成了为期2年的治疗。127例患者中，最终可用于评价疗效者四例，其中17只雌二醇0025mg组23例、0．05mg组23例、0．ling组别例2安慰剂组32例。1年后与安慰剂组对比，治疗组（种剂量）的腰椎骨矿质密度显著优于安慰剂组，尤以0．025mg组为著（P＝0．054）。2年后，治疗组的骨质密度也与安慰剂组有显著差异。17开雌H醇治疗2年后，…     

Microbiological Synthesis of 17β-Hydroxyandrosta-1,4-dien-3-one

Pharmaceutical Chemistry Journal -     

碘.β—环糊精包合物的制备

本文研究了碘·β-环糊精(I_1-β-CD)包合物的制备方法,经差示扫描量热法证实所制得的物质为包合物,并将I_1-β-CD包合物制备成了胶囊剂和混悬剂。     

真菌Nigrospora sp.Z18-17的化学成分

目的研究Nigrospora sp.Z18-17号菌株代谢产物,对其化学成分进行分离鉴定。方法采用硅胶柱色谱、凝胶柱色谱、HPLC等方法进行分离纯化,通过理化性质和波谱分析确定化合物的结构。结果从Nigrospora sp.Z18-17菌株发酵液和菌丝体中分离得到7个化合物,分别鉴定为:2-羟基-3-(4-羟基苯基)丙酸甲酯[2-hydroxy-3-(4-hydroxyphenyl)propionic acid methyl ester,1]、对羟基苯乙醇α-羟基丙酸酯(4-hydroxyphenethyl 2-hydroxypropanoate,2)、环(脯-缬)[cyclo(Pro-Val),3]、环(脯-脯)[cyclo(Pro-Pro),4]、环(苯丙-脯)[cyclo(Phe-Pro),5]、环(酪-羟脯)[cyclo(Tyr-Hyp),6]、尿嘧啶核苷(uridine,7)。结论化合物1～7均为首次从Nigrospora sp.Z18-17号菌株的代谢产物中分离得到。     

17α-Alkynyl 3α, 17β-androstanediol non-clinical and clinical pharmacology, pharmacokinetics and metabolism

17α-ethynyl-5α-androstane-3α, 17β-diol (HE3235, Apoptone) is an orally bioavailable synthetic analogue of 3β-androstanediol, that is active in rodent models of prostate and breast cancer, and is in Phase IIa clinical trials for the treatment of early- and late-stage prostate cancer. In preparation for clinical studies, nuclear hormone receptor and P450 interactions for HE3235 and major metabolites were characterized in vitro, and pharmacokinetics and metabolite profiles were studied in rodents, dogs, and monkeys. Four-week safety studies conducted in rats and dogs indicated a substantial margin of safety for clinical use, and no evidence of electrocardiographic or neurological effects, although anorexia, thrombocytopenia, and hypokalemia were identified as potentially dose-limiting toxicities at superpharmacological exposures. Pharmacokinetics and drug metabolism have been studied in prostate cancer patients.     

Geranylgeraniol and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester isolated from Pterodon pubescens Benth.: Further investigation on the antinociceptive mechanisms of action

The crude alcoholic extracts obtained from Pterodon pubescens Benth. seeds are widely used in Brazilian folk medicine as anti-inflammatory, analgesic, anti-rheumatic tonics and depurative preparations. We previously demonstrated the antinociceptive activity on writhing capsaicin, glutamate, and hot-plate tests of two compounds isolated from P. pubescens: geranylgeraniol (C1) and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester (C2). This work is a continuation of the previous study investigating the possible mechanisms of action for compounds C1 and C2, and the differences between them. The present study demonstrated that when administered intraperitoneally (i.p.): i), compounds C1 and C2 produced significant anti-allodynic activity during the acute phase of the Complete Freund's Adjuvant (CFA)-induced persistent pain model; ii) compound C1 produced significant anti-hypernociception activity in the carrageenan-induced pain model; iii) compound C2 presented a significant loss of activity after p-chlorophenylalanine methyl ester hydrochloride (PCPA) [5-HT synthesis inhibitor] treatment, suggesting that the mechanisms of action could be related to either the synthesis or release of serotonin; iv) compound C1 presented a significant loss of activity after ondansetron (5-HT(3) receptor antagonist) treatment suggesting activity upon 5-HT(3) serotonin receptors; v) compound C1 presented a significant loss of activity after efaroxan (mixed I(1) imidazoline/α(2)-adrenoceptor antagonist) treatment suggesting the participation of this compound upon imidazoline I(1) receptors; and vi) both compounds C1 and C2 did not appear to exert their activity via 5-HT(1A), 5-HT(2A), imidazoline I(2), α(2)-adrenoceptor, nitric oxide, GABA(A), acetylcholine muscarinic, and nicotinic receptors when evaluated in acetic acid-induced nociception.