Novel progesterone receptor modulators with gene selective and context-dependent partial agonism

Progesterone receptor (PR) modulators are used in contraception and post-menopausal hormone therapy, and are under clinical development for reproductive disorders such as uterine fibroids and endometriosis. Development of tissue selective PR modulators (SPRMs) with reduced side effects and improved pharmacology represents a large unmet medical need in the area of women’s health. One approach to addressing this need is to focus on the two PR isoforms PR-A and PR-B. In vitro and in vivo studies have revealed both distinct as well as overlapping gene regulation and functional responses of the two PR isoforms that suggests that PR-A selective modulators may retain a desired biological profile. We have identified a chemical series of 4-(4-chlorophenyl)-substituted piperazine carbimidothioic acid esters (PCEs) that have partial PR agonist activity and selectively activate some PR-A isoform regulated genes in T47D cells. However, full microarray analysis in these cells does not predict a global isoform selective profile for these compounds, but rather a unique gene-selective profile is observed relative to steroidal progestins. Using multiplexed peptide interaction profiling and co-activator recruitment assays we find that the mechanism of partial agonism is only partly defined by the ability to recruit known co-activators or peptides but also depends on the cell and promoter context of the gene under investigation. The data demonstrate global consequences of mechanistic and functional differences that can lead to selective biological responses of novel steroid receptor modulators.     

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目的研究中国乳腺癌病人术后死亡风险时间分布规律。方法回顾性研究1992年1月至2003年12月复旦大学附属肿瘤医院手术治疗并且资料完整的原发性乳腺癌2214例临床资料,对其进行生存及死亡风险分析。结果单因素和多因素生存分析均提示肿瘤大小、腋下淋巴结转移状态和孕激素受体状态是影响总存活率的预后因素(P<0.05)。全组病人年死亡风险曲线呈现双峰型,死亡高峰分别位于术后第2年及术后第9.5～10年,且该死亡模式在肿瘤较大及淋巴结阳性的亚组中更为明显。激素受体状态虽不改变病人的双峰型复发模式,但却呈现出一定的时间依赖性。结论乳腺癌术后死亡风险时间分布呈现出一定的规律,针对性制定术后随访和辅助治疗的新策略,可能有助于改善乳腺癌的治疗效果。     

Progesterone Receptor in Patients with Hepatolithiasis

Altered gallbladder motility by progesterone has been recognized as an important factor in the development of gallstones. There are two types of hepatolithiasis, that occurs de novo in the intrahepatic ducts with an intact gallbladder (primary hepatolithiasis) and that which originates in the gallbladder and the stones migrate into the intrahepatic duct (secondary hepatolithiasis). This study was designed to evaluate the possible role of the progesterone receptor of gallbladder in the pathogenesis of hepatolithiasis. Eighty-four patients with hepatolithiasis (34 patients had primary hepatolithiasis and the other 50 patients secondary hepatolithiasis) were included. Paraffin-embedded specimens of gallbladder were processed to have an immunohistochemical staining for progesterone receptor. Positivity for progesterone receptor of gallbladder specimens was noted in eight patients (23.5%) with primary hepatolithiasis and in 23 patients (46%) with secondary hepatolithiasis. There is significant difference (P = 0.031) in positive rate for progesterone receptor between the primary hepatolithiasis and secondary hepatolithiasis groups. In conclusion, many more patients (46%) with secondary hepatolithiasis show positivity for progesterone receptor of gallbladder specimens than patients with primary hepatolithiasis (23.5%) (P = 0.031). This phenomenon is intriguing and should be of further evaluation and elucided.     

Pharmacokinetics and Tolerability of a Novel 17β-Estradiol and Progesterone Intravaginal Ring in Sheep

This study reports the preparation, in vitro release, pharmacokinetics, and local tolerability of novel ethylene-vinyl acetate intravaginal rings (IVRs) delivering 17β-estradiol (E₂) and progesterone (P), in drug-naïve ovariectomized female Dorset crossbred sheep. After preparation and assessment of in vitro release of E₂ and P, animals were randomized to treatment groups 1 or 2 (comparator rings releasing 50 or 100 μg/d E₂, respectively), groups 3 or 4 (ethylene-vinyl acetate IVRs, 160 μg/d E₂ with 4 [160/4 IVR] or 8 mg/d P [160/8 IVR], respectively), or group 5 (160 μg E₂ and 10 mg P administered intravenously). IVRs were placed on day 1 and remained in place through day 29. Animals underwent daily examinations to confirm ring placement, and vaginal irritation was scored from 0 (none) to 4 (severe). Blood samples were taken at scheduled times for pharmacokinetic analysis. Postmortem examinations performed on groups 1-4 were macroscopic and microscopic evaluations, including irritation scoring and histopathology. IVRs were retained over 28 days in all but 1 animal (group 4). In all animal groups, clinical observations showed no significant abnormal findings. Pharmacokinetic analysis in the animals showed sustained release of E₂ and P over a 28-day period. Irritation scores and microscopic assessments were consistent with foreign object placement. A novel 2-drug IVR delivery system was well tolerated in a sheep model and pharmacokinetic release was as expected over a 28-day release period. These results will guide future human clinical studies.     

不同乳腺癌细胞株雌激素和孕激素受体的Western blot检测

目的　建立人类雌激素受体 (estrogenreceptor,ER)和孕激素受体 (progesteronereceptor,PR)Westernblot检测方法 ,并对不同乳腺癌细胞株ER、PR表达情况进行测定。方法　取不同乳腺癌细胞株对数生长期细胞提取总蛋白 ,分别用ERα单克隆抗体 1D5和PR单克隆抗体PgR6 36以Westernblot方法对ER、PR的表达情况进行检测 ,并与同期固定细胞的流式细胞术 (FCM)检测结果进行比较。结果　T 4 7d、MCF 7、ZR 75 1细胞ERα的Westernblot检测可得到清晰且分子量正确的条带 ,T 4 7d、ZR 75 1细胞可见清晰且分子量正确的PR条带 ,同期固定细胞ER、PR阳性表达的FCM检测结果与Westernblot检测结果一致。结论　Westernblot检测ER、PR的方法是可行的。     

子宫内膜癌雌孕激素受体与临床病理分析

【目的】探讨子宫内膜癌雌激素受体 (ER)、孕激素受体PR)与临床、病理间的关系。【方法】对 83例子宫内膜癌患者手术标本常规进行病理检查 ,同时采用免疫组化技术检测ER、PR。【结果】子宫内膜癌组织ER、PR及ER和PR同时阳性率较高 ,分别为 5 6 6 %、81 9%、5 3 0 %。绝经前子宫内膜癌组织ER、PR阳性率高于绝经后 (P <0 0 5 )。子宫内膜癌Ⅰ期ER、PR阳性率高于Ⅱ期以上者 (P <0 0 5 )。子宫内膜癌组织高分化的ER阳性率高于中低分化 (P <0 0 5 ) ,而PR阳性率与组织分化程度相关不大 (P >0 0 5 )。【结论】子宫内膜癌ER、PR阳性表达与手术病理分期、分化程度有关 ,对指导术后内分泌治疗有一定价值。     

Hormonal therapy in ovarian cancer

Ovarian carcinoma continues to be the leading cause of death due to gynecological malignancy. Epidemiologic studies indicate that steroid hormones play roles in ovarian carcinogenesis. Gonadotropins, estrogen, and androgen may be causative factors, while gonadotropin-releasing hormone and progesterone may be protective factors in ovarian cancer pathogenesis. Experimental studies have shown that hormonal receptors are expressed in ovarian cancer cells and mediate the growth-stimulatory or growth-inhibitory effects of the hormones on these cells. Hormonal therapeutic agents have been evaluated in several clinical trials. Most of these trials were conducted in patients with recurrent or refractory ovarian cancer, with modest efficacy and few side effects. Better understanding of the mechanisms through which hormones affect cell growth may improve the efficacy of hormonal therapy. Molecular markers that can reliably predict major clinical outcomes should be investigated further in well-designed trials.     

补肾益气中药对小鼠围着床期子宫内膜形态结构及PR表达的影响

[目的]探讨补肾益气中药改善胚泡着床障碍小鼠着床期子宫内膜微环境的机制。[方法]于妊娠第4天9:00利用米非司嗣建立胚泡着床障碍小鼠模型,予补肾益气中药进行干预,于妊娠第4天21:00处死小鼠,观察子宫内膜形态学变化,采用免疫组化技术及Western blot技术检测子宫PR的表达。[结果]与正常组相比,模型组子宫内膜发育不良,中药组子宫内膜的发育与正常组相似;中药组子宫内膜腺体、间质、腔上皮细胞PR表达范围和强度及PR蛋白的表达均较模型组显著增高,子宫PR蛋白的表达较模型组显著增高,与正常组无显著差异;[结论]补肾益气中药通过改善子宫内膜发育,调节胚泡着床障碍小鼠子宫组织PR蛋白的表达,改善子宫内膜着床微环境,利于胚泡着床。     

Prospective randomized trial to evaluate the efficacy of a vaginal ring releasing progesterone for IVF and oocyte donation

A polysyloxane vaginal ring containing 1g of natural progesterone was developed as luteal supplementation for women treated with IVF-embryo transfer and for agonadal women participating in an oocyte donation programme. The ring provides continuous release of progesterone (10-20 nmol/l) for 90 days. The efficacy of this form of progesterone supplementation was evaluated in two multicentre prospective randomized trials. IVF-embryo transfer trial: After oocyte aspiration, 505 women were randomly allocated to progesterone supplementation with vaginal ring or i.m. progesterone (50 mg/day). The clinical pregnancy rate was 36.6% in both groups. Implantation rate was 15.9% in the vaginal ring and 16.0% in i.m. progesterone. Oocyte donation trial: After endometrial proliferation with micronized oestradiol, 153 women were allocated to progesterone replacement with a vaginal ring or i.m. progesterone (100 mg/day). Clinical pregnancy rate was 39.8 and 28.6% respectively. Implantation rate was significantly higher with the vaginal ring compared with i.m. progesterone (19.9 and 11.6% respectively, P = 0.006). The vaginal ring is a novel development which provides continuous release of progesterone for 90 days. In IVF-embryo transfer, its effectiveness is similar to daily i.m. injections. In oocyte donation the ring provides a progestative milieu which improves the implantation rate and eliminates the discomfort of daily i.m. injections.