阿扎斯丁和环氧司坦类似物的合成及其抗早孕活性

阿扎斯丁和环氧司坦能竞争性抑制3β-羟甾脱氢酶,干扰孕酮合成,从而具有抗早孕作用。本文设计合成了12个新的类似物,进行抗早孕筛选。其中6个中间体和3个副产物亦未见文献报道。本文提出的17α,18-二甲基-17-羟基雌甾-4-烯-3-酮(17)-锅合成法,总收率48%     

用回归设计法优选甾族沃氏氧化反应条件

沃氏氧化反应(Oppenauer Oxidation)在甾体药物的生产中有着广泛的应用。16,17α-环氧-孕甾-4-烯-3,20-二酮(简称沃氏氧化物),是甾体药物的一个重要的中间体。沃氏氧化物是由3β-羟基-孕甾-5,16-二烯-20-酮-3-醋酸酯(简称双烯),经双氧水氧化得3β-羟基-16,17α-环氧-孕甾-5-烯-20-酮(简称氧桥)再经沃氏氧化而生成的。     

甾体激素 Ⅱ.9α-氟副肾皮质激素的新合成路线

自16α,17α-环氧孕甾-4-烯-11α-醇-3,20-二酮(Ⅰ)經对甲苯磺酰化和消去反应得到16α,17α-坏氧孕甾-4,9(11)-二烯-3,20-二酮(Ⅱ),用氫溴酸打开环氧。氫解脫溴得到孕甾-4,9(11)-二烯-17α-醇-3,20-二酮(Ⅲ),引入C₂₁-OAc得到孕甾-4,9(11)-二烯-17α,21-二醇-3,20-二酮21-醋酸脂(Ⅳ),然后按已知方法合成9α-氟-可的唑(Ⅵ).(Ⅵ)对(Ⅰ)的收率为17.4%理論。也进行了由(Ⅲ)先引入9β,11β-环氧,然后引入C₂₁-OAc以合成9α-氟可的唑的試探。     

5α-△~(9(11))-16β-甲基-3β,17α,21-三羟基-孕甾烯-3β,21-双醋酸酯-20酮和5α,17α-甲基-17β-羟基-雄甾-3酮的微生物转化

用诺卡氏菌与节杆菌混合菌种转化从蕃麻皂素制得的中间体5α-△~((?)(11))-16β-甲基-3β,17α,21三羟基孕甾烯-3β,21-双醋酸酯-20酮(Ⅰ)得50％的16β-甲基-△~(1,4,9(11))-孕甾三烯-20酮(Ⅱ)和少量的16β-甲基-9,11α环氧-△~1,4孕甾二烯-20酮(Ⅲ)。另外,又用同样的混合菌种转化从剑麻皂素制得的中间体5α,17α甲基-17β羟基-雄甾-3酮(Ⅳ)得50％17α甲基-17β羟基-△~(1,4)-雄甾二烯-3酮(Ⅴ)。如改变培养基则得3,17β-羟基-17α-甲基-9酮基-9,10开环-1,3,5(10)雄甾三烯化合物。     

螺内酯的合成

雄烯二酮和原甲酸三乙酯反应后用碘化三甲基锍环氧化得到17β,20β-环氧-3-乙氧基-17α-孕甾-3,5-二烯,然后经与丙二酸二甲酯成内酯环、NBS溴化后脱溴化氢,与硫代乙酸加成制得利尿药螺内酯,总收率57%.     

17β-羟基-4ɑ,5ɑ-环氧雄甾烷并[2,3-d]异噁唑的合成工艺改进

17β-羟基雄甾-4-烯-3-酮与甲酸乙酯在甲醇钠条件下经缩合生成17β-羟基-2-羟亚甲基雄甾-4-烯-3-酮,与盐酸羟胺环合形成17β-羟基-4-烯-雄甾烷并[2,3-d]异噁唑,最后经m-CPBA环氧化制得曲洛司坦关键中间体17β-羟基-4ɑ,5ɑ-环氧雄甾烷并[2,3-d]异噁唑,总收率为70%,纯度为99.1%。     

醋酸乌利司他的合成

3,3-(亚乙二氧基)雌甾-5(10),9(11)-二烯-17-酮经乙炔加成,与苯硫氯反应,脱苯亚磺酰基,去甲基重排,羰基保护制得3,3:20,20-二(亚乙二氧基)-17α-羟基-19-去甲孕甾-5(10),9(11)-二烯(7),7以六氟丙酮为催化剂环氧化制得3,3:20,20-二(亚乙二氧基)-17α-羟基-5α,10α-环氧-19-去甲孕甾-9(11)-烯,然后经格氏加成、去保护、乙酰化合成醋酸乌利司他,总收率23.1%,纯度99.8%。优化后工艺生产成本低,生产周期短,操作简便。     

用根霉80322进行16α,17α-环氧-△~4-孕甾烯-3,20-二酮11α羟基化的副产物

11α-羟基-16α、17α-环氧-△~4-孕甾烯-3,20-二酮(Ⅱ)是合成可的松、强的松、肤轻松等甾体激素药物的重要中间体,由16α,17α-环氧-△~4-孕甾烯-3,20-二酮(Ⅰ)经霉菌氧化而得。上海工业微生物研究所从400余株野生梨的霉菌中,选出一株根霉80322新菌株,可在2％底物投料浓度,37℃发酵36～48h,使转化率达到55％以上,其主要发酵产物除11α-羟基-16     

醋酸乌利司他合成路线图解

醋酸乌利司他(ulipristal acetate,1),化学名为17α-乙酰氧基-11β-(4-二甲胺基苯基)-19-去甲孕甾-4,9-二烯-3,20-二酮,由法国HRA制药公司研究开发,2009年5月获得欧盟委员会批准上市,商品名EllaOne,可用作妇女无保护性交或避孕失败后5天内使用的紧急避孕药.     

孕二烯酮有关物质的合成

目的为更好地对孕二烯酮原料药质量进行控制,合成孕二烯酮原料药中的2种有关物质。方法以13β-乙基-15α-羟基雌甾-4-烯-3,17-二酮为起始原料,经过3位羰基缩酮化保护、17位乙炔化和水解脱保护基共3步反应,得到有关物质13β-乙基-15α,17β-二羟基-18,19-双去甲基-17α-孕甾-4-烯-20-炔-3-酮,收率为12.1%;该有关物质经乙酰化得到另一种有关物质13β-乙基-17β-羟基-15α-乙酰氧基-18,19-双去甲基-17α-孕甾-4-烯-20-炔-3-酮,收率为72.9%。结果与结论合成了孕二烯酮原料药中的2种有关物质,并经1H-NMR、13C-NMR和MS谱确证结构,2种有关物质的纯度经HPLC检测均在98%以上,可以作为孕二烯酮原料药质量控制的对照品。     

醋酸棉酚对hCG促进大鼠分散黄体细胞产生孕酮的影响

本文在体外研究了醋酸棉酚对分散大鼠黄体细胞产生孕酮的影响及其作用机制。当棉酚浓度在20μg/ml、作用时间1h,hCG刺激黄体细胞孕酮的生成量显著下降,但对基础孕酮的产生无明显影响;上述剂量的棉酚在作用4h后时细胞存活率无明显影响。经棉酚作用过的黄体细胞,经2h再孵育,不能恢复其对hCG的反应性。同时棉酚显著降低黄体细胞cAMP产生。结果提示,棉酚可能通过降低cAMP产生而抑制hCG的生孕酮作用。     

盐酸苯乙哌啶和dl-15甲基PGF2α甲酯对大鼠卵巢黄体细胞的影响

盐酸苯乙哌啶(R1132)10μg/ml或dl-15甲基PGF_2α甲酯(PG05)5或10μg/ml在体外能明显抑制黄体细胞对hCG的反应性,使孕酮分泌下降。假孕大鼠po R1132 10 mg/kg或Sc PG 05 5.1 mg/kg不影响卵巢孕酮分泌,合并给药后却能使其降低。R1132无抗孕酮作用。卵巢分泌孕酮减少可能是抗早孕的主要原因.假孕大鼠po R1132 50 mg/kg或sc PG050.5 mg/kg可抑制卵巢腺苷环化酶的活性.该酶可能是R1132或PG05在大鼠抗早孕作用的重要靶酶。     

Effects of cadmium on preimplantation and early postimplantation mouse embryos in vitro with special reference to their trophoblastic invasiveness

Cadmium chloride, at concentrations of 0.5 or 1 microgram/ml medium, did not affect the trophoblastic invasiveness of mouse embryos treated for 24 hours at 4-cell and morula stages. At higher concentrations of 5 or 10 micrograms/ml medium, most treated embryos in vitro underwent degeneration while a few survivors formed trophoblastic outgrowths with variable areas. Cadmium chloride, at a low concentration of 0.5 microgram/ml medium presented continuously to blastocysts after attachment in vitro, has significantly retarded the trophoblastic outgrowth areas and reduced the number of trophoblastic giant-cell nuclei, though the spreading blastocysts appeared morphologically normal. At higher concentrations of 1 or 5 micrograms/ml medium, cytoplasmic disintegration and detachment of trophoblasts were observed. It is suggested that cadmium may interfere with the cell division and/or the transformation of trophectoderm cells into giant cells, resulting in the retardation of the trophoblastic outgrowths.     

In vitro embryotoxicity of petroleum creosote monitored via mouse preimplantation embryo culture.

A mouse preimplantation embryo culture system was utilized to characterize the in vitro embryotoxicity of petroleum creosote (PC), a complex mixture of aliphatic and polycyclic aromatic hydrocarbons. ICR mouse embryos, collected on d 3.5 of gestation (blastocyst stage), were exposed for 1 h to varying concentrations of petroleum creosote in serum-supplemented culture medium. Parallel embryo cultures were exposed to PC in medium supplemented with rodent hepatic S9 microsomal fractions to monitor the role of bioactivation in PC-induced embryotoxicity. Embryos were subsequently cultured in control medium for 72 h and observed for viability as well as specific, time-dependent developmental end points--hatching and attachment to the culture dish at 48 h, and trophoblastic outgrowth with a distinct inner cell mass at 72 h. Embryonic viability varied in inverse proportion to PC concentration. Petroleum creosote caused embryolethal effects at concentrations of 33 micrograms/ml of culture medium and 54 micrograms/ml. Embryotoxicity was not observed at 22 micrograms/ml. Culture supplementation with rodent hepatic S9 fractions did not modify, either qualitatively or quantitatively, the embryotoxicity of PC in vitro. These findings implicate PC as a prenatal toxicant and support environmental and human health concerns regarding PC exposure from PC-containing chemical waste sites.     

In vitro teratogenicity of acetylsalicylic acid on rat embryos: studies with various culture conditions

Rat embryos taken at day 9.5 of gestation were exposed in vitro to acetylsalicylic acid (aspirin) using various culture conditions. It was observed that embryos were sensitive to aspirin emulsified in olive oil at concentrations greater than or equal to 150 micrograms/ml. Between 43% and 66% of the embryos exhibited multiple malformations depending on the culture medium, 100% homologous rat serum or Waymouth medium supplemented with 50% rat serum, respectively. At concentrations greater than or equal to 400 micrograms/ml aspirin induced further toxic effects on embryo growth and differentiation. When gelatin was used as the drug-delivery system, aspirin at concentrations of greater than or equal to 150 micrograms/ml induced some malformations (mainly irregular somite shapes) in 57% of the embryos cultured in Waymouth medium, but in only 13% of the embryos grown in 100% serum. At concentrations which were greater than or equal to 400 micrograms/ml aspirin induced dysmorphogenic effects in all embryos, without any concomittant toxicity.     

一些保肝药物对原代培养大鼠肝细胞糖原合成功能的影响

本文参照PO Seglen的方法并加以修改,建立了原代培养大鼠肝细胞糖原合成功能的测定体系。观察到联苯双酯既能使正常肝细胞合成糖原增加88%,又能保护肝细胞完全拮抗四氯化碳对其功能的损伤;银耳多糖能使四氯化碳对肝细胞糖原合成功能的损伤减轻57%;去甲斑蝥素10μg/ml能增加肝细胞糖原合成,浓度增加到100μg/ml时,此作用减弱,1000μg/ml则明显抑制糖原的合成,而且在10～100μg/ml浓度时,即能加强四氯化碳的损伤作用;100μg/ml CL₁₅₀₀和熊果酸二钠单独应用可增加肝细胞糖原合成,但与四氯化碳同时应用,反而加重对糖原合成的抑制作用。     

醋酸棉酚对离体大鼠黄体和人蜕膜、滋养层细胞的影响

以细胞分泌功能和存活率为指标,观察了醋酸棉酚对无血清培养的大鼠黄体和人蜕膜、滋养层细胞的直接损伤作用和可能的作用途径。结果表明:醋酸棉酚对黄体细胞有直接杀伤作用,LD₅₀为:1.6(0.4～2.9)μg·mL^-1。在非致死剂量(0.5μg·mL^-1)下,醋酸棉酚显著抑制黄体细胞基础孕酮分泌,但不能显著抑制hCG,Forskolin的促孕酮作用和3β-HSD的活性。此外,醋酸棉酚对蜕膜细胞和滋养层细胞也有直接杀伤作用,其LD₅₀分别为:3.5(0.4～6.6)μg·mL^-1,4.1(0.6～7.6)μg·mL^-1。本文结果提示,直接杀伤黄体,蜕膜和滋养层细胞并影响孕酮合成是醋酸棉酚抗生育的重要作用环节。     

Sensitivity of preimplantation mouse embryos to abrin

The effect of the plant toxin abrin on preimplantation mouse embryos in vitro was studied. Two-cell embryos from C57BL/6J or B6CBA/F1/Bom female X B6CBA/F1/Bom male mice were exposed to abrin for 72 hrs in vitro, in medium containing abrin in concentrations ranging from 0.1 to 10,000 pg/ml. Two-cell mouse embryos were also exposed to corresponding concentrations of ricin in vitro. The effect of abrin was evaluated by daily morphological observation of the embryos up to the blastocyst stage and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. The effect of ricin was evaluated by morphological observation of the embryos up to the blastocyst stage. The results showed that 2-cell mouse embryos were highly sensitive to abrin, 1 pg/ml being the lowest concentration of the toxin that significantly inhibited the development of 2-cell embryos to the blastocyst stage. With ricin statistically significant inhibition of blastocyst formation was obtained at a concentration of 10 pg/ml. The ID50 for abrin was calculated to be 24 pg/ml and for ricin 47 pg/ml. A dose-dependent lag period was observed before the effect of abrin or ricin on the formation of blastocysts became evident. Incorporation of 3H-thymidine, 3H-uridine and 14C-leucine into DNA, RNA and protein, respectively, was also inhibited by abrin after different time intervals in culture.     

Deciduoma formation in rats treated neonatally with the anti-estrogens, tamoxifen and MER-25

Female rats of the T strain were given single daily injections of 100 micrograms and 200 micrograms of tamoxifen (Tx) and MER-25 (MER) for 5 days beginning on the day of birth (DAY 1). When sacrificed on Day 60, the Tx-treated rats (Tx rats) exhibited continued vaginal diestrus, whereas the females given MER or the vehicle alone showed regular estrous cycles. Ovaries from Tx rats were polyfollicular without corpora lutea, while those from MER rats, as well as from the controls given the vehicle alone, invariably contained both follicles and corpora lutea. In Tx rats, the uteri underwent atrophy, containing few uterine glands in an endometrium largely occupied by fibroblasts. Decidual response of the uterus to intraluminal oil instillation was markedly reduced in Tx rats given an appropriate regimen of progesterone and estradiol injections following ovariectomy on Day 60. By contrast, MER given neonatally had little effects on decidualization. Since ovariectomy on Day 10 brought about no amelioration of the decidualization in Tx rats, it is suggested that the lowered deciduogenic responsiveness to the instillation was caused by a direct action of Tx on the uterus of neonatal rats rather than by a Tx-induced alteration of hypothalamo-hypophyseal-ovarian system. Differences in effect on the female reproductive system between Tx and MER were discussed.