contraceptives--postcoital and anordiol

Anordiol, the dihydroxylated metabolite of anordrin, is an antiestrogen with estrogenic activity that is known to inhibit fertility. The following study was conducted to determine the mechanism of this antifertility effect. Anordiol was administered orally to rats, prior to implantation, on Day 2 of pregnancy. Control animals were treated with the vehicle only. The effectiveness of the agent in terminating pregnancy was determined on Day 14 of pregnancy. Anordiol was 100% effective in abolishing pregnancy at a dose of 0.6 mg/Kg. Administration of smaller doses resulted in a decreased number of implanting embryos, in a dose-dependent manner. An additional dose of anordiol on Day 3 of pregnancy yielded similar results. To determine whether pregnancy impairment by anordiol is exerted via the embryo or via the uterus, reciprocal embryo transfers were performed. Day 5 blastocysts were transferred into the uteri of pseudopregnant rats. In one set of experiments, the donor rats were treated with anordiol, and in the second set the recipient rats were treated. The results indicate that the effects of anordiol administration are exerted via the embryo as well as the uterus.

Topics: Abortion, Induced; Animals; Blastocyst; Contraceptives, Postcoital; Embryo Implantation; Embryo Transfer; Embryo, Mammalian; Female; Gestational Age; Norandrostanes; Pregnancy; Rats; Rats, Wistar; Uterus

The effectiveness of mifepristone, onapristone, and ORG 31806 alone or in combination with anordiol to terminate pregnancy in the rat was evaluated. ORG 31806 at a dose of 2 mg/kg/day, mifepristone at 4 mg/kg/day, and onapristone at 8 mg/kg/day, terminated pregnancy in all treated animals. Anordiol, an antiestrogen, at a dose of 5 mg/kg/day, terminated pregnancy in all treated animals. Anordiol acted synergistically with all three antiprogestins terminating pregnancy in the rat. The antiprogestins at doses that were either partially effective or non-effective became 100% effective when administered with a non-effective dose of anordiol. Thus, combination of ORG 31806 (1 mg/kg/day) plus anordiol (0.31 mg/kg/ day), mifepristone (1 mg/kg/day) plus anordiol (0.62 mg/ kg/day), and onapristone (2 mg/kg/day) plus anordiol (2.5 mg/kg/day) terminated pregnancy in all treated animals. These combinations of the antiprogestins and anordiol decreased significantly the serum progesterone levels but not serum 17 beta-estradiol levels. The present results indicate that the most potent combination was ORG 31806 plus anordiol.. The pregnancy termination potency of varying doses of mifepristone, onapristone, and ORG 3806--alone and in combination with the estrogenic/antiestrogenic compound anordiol--was evaluated in adult rats. The antiprogestins and anordiol alone were administered to pregnant female rats on days 7, 8, and 9 of pregnancy and the presence or absence of embryos in utero was determined on day 16. ORG 31806 at a dose of 2 mg/kg/day, mifepristone at 4 mg/kg/day, and onapristone at 8 mg/kg/day terminated pregnancy in 100% of animals; 5 mg/kg/day of anordiol was required. Anordiol acted synergistically with all three antiprogestins. Antiprogestin doses that were either partially effective or ineffective became 100% effective when administered with a noneffective dose of anordiol. The combination of ORG 31806 (1 mg/kg/day) and anordiol (0.31 mg/kg/day) had the most potent pregnancy termination activity. The administration of antiprogestins in combination with anordiol at doses that effectively terminate pregnancy was associated with a significant, persistent reduction in serum progesterone, but no change in serum estradiol levels. The effectiveness of ORG 31806 and anordiol in terminating pregnancy should be evaluated in a non-human primate model to determine its potential clinical use.

Topics: Abortion, Induced; Administration, Oral; Animals; Cohort Studies; Contraceptives, Postcoital; Dose-Response Relationship, Drug; Drug Combinations; Drug Synergism; Estradiol; Estrenes; Female; Furans; Gonanes; Hormone Antagonists; Male; Mifepristone; Norandrostanes; Pregnancy; Progesterone; Rats; Rats, Sprague-Dawley

In the present studies, we observed the regulation of water channel gene (AQP-CHIP) expression by estradiol (E2) and anordiol, an antiestrogen with agonist activity, in immature female rat uterus. Antisense and sense oligonucleotide primers corresponding to the consensus sequences of two rats AQP-CHIP water channels were synthesized and used to amplify a cDNA fragment that was reverse-transcripted from rat uterine total RNA preparation. E2 administered as a single dose of 40 micrograms.kg-1 to immature female rats induced a significant increase in AQP-CHIP mRNA expression 9 h after treatment. The lowest effective doses of E2 and anordiol were 40 and 50 micrograms.kg-1, respectively. The stimulatory effect of anordiol was more pronounced than that of E2. The present results suggest that AQP-CHIP water channel gene expression may be involved in E2- and anordiol-mediated water imbibition and luminal fluid production in the uterus.

Topics: Animals; Aquaporin 1; Aquaporins; Contraceptives, Postcoital; Estradiol; Female; Gene Expression; Norandrostanes; Rats; Rats, Sprague-Dawley; RNA, Messenger; Uterus

A 250 kDa secretory protein was isolated from the uterine luminal fluid (ULF) obtained from estradiol-treated ovariectomized rats. Antiestrogens blocked the production of this protein. The protein components were separated and purified by SDS-PAGE. Polyclonal antibodies were raised against the 250 kDa protein and used to identify the protein by Western blot. 17 beta-estradiol (E2) at a dose of 0.005 mg/kg/d administered s.c. for 3 days to ovariectomized rats stimulated a marked increase in the production of the 250 kDa protein. Anordiol at a low dose of 5 mg/kg/d x 3 p.o. or 0.25 mg/kg/d x 3 s.c. stimulated the production of the 250 kDa protein. Treatment with higher doses (10 mg/kg/d x 3 p.o., or 1.25 mg/kg/d x 3 s.c.) was less effective in inducing production of this protein. Also anordiol partially inhibited the stimulatory action of E2; whereas ICI 182,780, a pure antiestrogen, at a dose of 0.3 mg/kg/d x 3 s.c. completely blocked the stimulatory action of E2. The 250 kDa protein was not detected in the blood obtained from E2-treated ovariectomized rats. The anti-complement C3 and anti-alpha 2-macroglobulin antibodies did not react with the ULF 250 kDa protein. The present results show that the production of the ULF 250 kDa protein is regulated by estradiol and is not a component of blood plasma. It is proposed that the estrogen-responsive 250 kDa protein may be involved in maintaining the viability of the fertilized ova and in the implantation of the blastocyst.

Topics: Animals; Blotting, Western; Contraceptives, Postcoital; Electrophoresis, Polyacrylamide Gel; Estradiol; Estrogen Antagonists; Female; Fulvestrant; Glycoproteins; Molecular Weight; Norandrostanes; Ovariectomy; Rats; Rats, Sprague-Dawley; Serpins; Uterus

The in vitro effect of anordrin and anordiol on the development of mouse two-cell embryos was studied.. Female mice were primed with gonadotropins for superovulation and caged with male mice. Preimplantation embryos, at the two-cell stage, were recovered from the oviducts at 40 hr post-hCG. In the first experiment, two-cell embryos were exposed to culture medium containing different concentrations of anordrin for 3, 12, 24, and 80 hr and then grown in the anordrin-free culture medium and assessed for the formation of total and hatching blastocysts at 80 hr. In the second experiment, two-cell embryos were grown in culture medium containing different concentrations of anordiol and assessed for the formation of total and hatching blastocysts at 80 hr in vitro.. Exposure of two-cell embryos to anordrin concentrations of 2.5-7.5 micrograms/ml for 12 hr, 2.5-5.0 micrograms/ml for 24 hr, and 2.5 micrograms/ml for 80 hr caused significant inhibition of the formation of total blastocysts and to 2.5-7.5 micrograms/ml for 12 hr, 1.0-2.5 micrograms/ml for 24 hr, and 1.0 micrograms/ml for 80 hr caused significant inhibition of the formation of hatching blastocysts, in a exposure time-dependent and dose-dependent manner. Exposure of two-cell embryos to anordiol concentrations of 15-25 micrograms/ml for 80 hr caused significant inhibition of the formation of total blastocysts and to 15-20 micrograms/ml for 80 hr caused significant inhibition of the formation of hatching blastocysts in a dose-dependent manner.. Anordrin and its metabolite anordiol inhibit the development of two-cell embryos in vitro.

Topics: Animals; Contraceptives, Postcoital; Embryo, Mammalian; Embryonic and Fetal Development; Embryonic Development; Female; Male; Mice; Norandrostanes; Organ Culture Techniques; Pregnancy; Time Factors

To develop a better postcoital contraceptive, the following antiestrogens were tested for their anti-implantation activity in the rat: anordrin, anordiol, tamoxifen, ICI 182,780, and RU 39411. The compounds were administered orally or subcutaneously (s.c.) to female rats on days 1, 2, and 3 of pregnancy. All the antiestrogens tested were 100% effective in preventing blastocyst implantation. The lowest effective doses when administered orally were 10, 1.25, 0.062, 6.0 (partially effective), and 0.01 mg/kg/day, respectively. The estimated median effective doses (ED50) were 5.60, 0.40, 0.035, 5.40, and 0.0074 mg/kg/day, respectively. When administered s.c., the minimum effective doses in preventing blastocyst implantation in all animals were 2.0, 0.1, 0.1, 0.1, and 0.01 mg/kg/day, respectively. Anordrin, anordiol, and ICI 182,780 were more potent when administered s.c.; whereas tamoxifen and RU 39411 were effective at similar doses when administered parenterally or orally. RU 39411 was the most potent among the antiestrogens tested and should be evaluated as a potential postcoital contraceptive. The administration of mifepristone, an antiprogestin, at a dose of 8 mg/kg/day blocked blastocyst implantation in all treated animals; whereas at a dose of 4 mg/kg/day or lower, the drug was ineffective. These findings confirm that estradiol and progesterone are essential for blastocyst implantation in the rat. The capacity of mifepristone to potentiate the anti-implantation activity of the antiestrogens was also determined. The combination of a non-effective dose of each of the antiestrogens (anordrin, anordiol, and tamoxifen), and RU 39411, with mifepristone at a non-effective dose, prevented pregnancy, demonstration that an antiprogestin and antiestrogen act synergistically in blocking blastocyst implantation in the rat. The antiestrogen compounds whose anti-implantation activities were potentiated by mifepristone were found to possess significant estrogenic activity, when assayed by measuring the increase in the uterine weights of ovariectomized rats. The only exception was ICI 182,780, which showed no estrogenic activity in the uterine weight bioassay and did not act synergistically with mifepristone in blocking blastocyst implantation. Estradiol was effective in preventing pregnancy at a dose of 1 microgram/kg/day. The combination of non-effective doses of estradiol and mifepristone did not prevent pregnancy. The findings that mifepristone potentiates the anti-. In New York, female and male rats copulated on the afternoon of proestrus as part of a study aiming to determine whether antiestrogens alone or in combination with mifepristone (RU-486) will block blastocyst implantation in the rat. This study is part of research efforts to develop a better postcoital contraceptive. The antiestrogens included RU39411; ICI 182,780; anordrin; anordiol; tamoxifen; and estradiol. The laboratory researchers treated the rats orally or subcutaneously on days 1, 2, and 3 of pregnancy. They killed them on day 8-9 to examine the uteri for the presence or absence of implanted embryos. All the antiestrogens effectively prevented blastocyte implantation. Using the measurement mg/kg/day, the lowest effective oral dose was 10 for anordrin; 1.25 for anordiol; 0.062 for tamoxifen; 6 (80% effective) for ICI 182,780; and 0.01 for RU 39411. These antiestrogens' estimated median effective doses were 5.6, 0.4, 0.035, 5.4, and 0.0074 mg/kg/day, respectively. In all animals, the minimum effective doses administered subcutaneously were 2, 0.1, 0.1, 0.1, and 0.01 mg/kg/day, respectively. Anordrin, anordiol, and ICI 182,780 were more effective during subcutaneous administration while tamoxifen and RU 39411 were as effective at similar doses during parenteral or oral administration. The most potent antiestrogen was RU 39411. This antiestrogen should be evaluated as a potential postcoital contraceptive. An 8 mg/kg/day dose of RU-486 blocked blastocyst implantation in all rats. The 4 mg/kg/day dose was completely ineffective. RU-486 augmented the activity of anordrin, anordiol, tamoxifen, and RU39411 synergistically, resulting in prevention of pregnancy at non-effective doses of RU-486 and the antiestrogens. These same antiestrogens also exhibited significant estrogenic activity as determined by an increase in uterine weights of ovariectomized rats. Estradiol prevented pregnancy at a dose of 1 mcg/kg/day. RU-486 did not potentiate estradiol. These findings suggest that the synergistic effect of anordrin, anordiol, tamoxifen, and RU39411 may be unique to these antiestrogens.

Topics: Administration, Oral; Animals; Contraceptives, Postcoital; Embryo Implantation; Estradiol; Estrogen Antagonists; Female; Fulvestrant; Injections, Subcutaneous; Male; Mifepristone; Norandrostanes; Pregnancy; Rats; Rats, Sprague-Dawley; Tamoxifen; Uterus

Anordiol (2 alpha,17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta,17 beta-diol) has been variously characterized as an estrogen and as an antiestrogen. To more completely understand the pharmacological properties of this contraceptive steroid, simultaneous responses were studied in uterine, vaginal, and hepatic tissues. Rats received 4 daily sc injections with either anordiol, clomiphene citrate (CC), or the vehicle alone (C+) starting on the first day of pseudopregnancy. Uteri were traumatized on day 4 of pseudopregnancy, and rats were sacrificed 5 days later. A pseudopregnant group without uterine trauma served as a negative control (C-). Mean uterine weights per animal and cytosolic estrogen (EcR) and progesterone (PcR) receptor activities per g of DNA were all 5- to 7-fold greater in the C+ group than in the other groups (all p < 0.05). However, anordiol and CC suppressed uterine weight without suppressing the stromal proliferative response; the DNA content of the uteri of anordiol- and CC-treated rats was similar to that of C+ rats. Vaginal tissue exhibited estrogenic responses to anordiol and CC with an increase in epithelial stratification compared to the C+ and C- groups even though no difference in levels of EcR/g of DNA were expressed 5 days after the last antiestrogen dose. Binding affinities and serum E2 and progesterone (P) concentrations were not statistically different among the groups. In conclusion, anordiol produced responses in the uterus and vagina of the pseudopregnant rat which were indistinguishable from those of CC, and, therefore, we conclude that anordiol acts on these tissues as an antiestrogen.

Topics: Animals; Clomiphene; Contraceptives, Postcoital; DNA; Estradiol; Estrogen Antagonists; Estrogens; Female; Liver; Norandrostanes; Organ Size; Progesterone; Pseudopregnancy; Rats; Receptors, Estrogen; Receptors, Progesterone; Uterus; Vagina

The purpose of this investigation was to determine the estrogenic and antiestrogenic activities of the potential contraceptive agent anordiol in the immature rat. A dose of 2.45 mumol of anordiol (AD), estradiol (E2), clomiphene citrate (CC), tamoxifen (TA), or the vehicle alone was administered to rats on the 25th and 29th days of age. Serum hormones were measured between 16:00 and 17:00 on days 30, 31, and 32; organ weights were determined on day 32. Anordiol and estradiol treatments significantly increased ovarian and uterine weight and serum LH concentrations, but CC and TA had no effect on these parameters. Vaginal cornification occurred before day 32 in all rats receiving anordiol or estradiol and in 3/5 and 4/5 rats receiving CC and TA, respectively, but not in control rats. Based on serum progesterone levels, ovulation was induced only in rats receiving anordiol or estradiol. All of the compounds tested significantly increased serum testosterone above levels in control animals, but both AD and E2 induced ovulation without a further increase in serum testosterone. We conclude that in the immature rat anordiol produces estrogenic responses in the vagina, uterus and in the hypothalamic-pituitary-ovarian axis, whereas TA and CC are estrogenic only on the vaginal and uterine epithelium.

Topics: Animals; Clomiphene; Contraceptives, Postcoital; Estradiol; Female; Genitalia, Female; Gonadal Steroid Hormones; Norandrostanes; Ovary; Ovulation; Radioimmunoassay; Rats; Rats, Sprague-Dawley; Tamoxifen; Uterus; Vagina

Because RU 486 synergizes with anordrin and its dihydroxylated metabolite to terminate established pregnancy in the rat and rabbit, the interactions of these agents were studied at two days postcoitally in the rat. RU 486 at a dose of 4 mg/kg did not prevent pregnancy when the animals were killed 12 days post insemination. A 2.5 mg/kg dose of anordrin prevented pregnancy in 14% of animals. By contrast, none of the animals became pregnant when treated with 2.5 mg/kg of anordiol. A non-effective dose of RU 486 (2 or 4 mg/kg) combined with a non-effective dose of anordrin (1.25 or 2.5 mg/kg) prevented pregnancy in all animals treated; there was no evidence of implantation sites or embryos when the animals were killed on day 12 post insemination. The same synergistic effect was observed when a small dose of RU 486 (e.g., 1 mg/kg) was combined with 0.6 mg/kg anordiol. To investigate the mechanism of pregnancy prevention, animals were treated two days postcoitally with 4 mg/kg of RU 486 plus 2.5 mg/kg of anordrin or 2 mg/kg of RU 486 plus 0.6 mg/kg of anordiol and were killed short intervals after treatment. These drugs had no effect by 6 h, but the numbers of embryos in oviducts were significantly reduced 12 h after treatment. By 24 h following treatment, no embryos were recovered from either the oviduct or the uterus. Progesterone and estradiol levels in serum collected 24 h after treatment were not significantly different from those of controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Contraceptives, Oral, Synthetic; Contraceptives, Postcoital; Drug Combinations; Drug Synergism; Embryo Implantation; Estradiol; Female; Male; Mifepristone; Norandrostanes; Ovum; Progesterone; Rats; Rats, Sprague-Dawley; Time Factors

We describe the chemical synthesis of the 2 beta-propionate-17 beta- hemisuccinate and 2 beta-hemisuccinate-17 beta-propionate diesters of anordiol (2 alpha,17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta,17 beta-diol) and the method for coupling them to bovine serum albumin and Affi-gel 102, in order to prepare antibodies for radioimmunoassay of anordrin. In addition, we describe the chemical synthesis of the following derivatives: 2 beta-ol-17 beta-propionate, 2 beta-propionate-17 beta-ol, 2 beta-hemisuccinate-17 beta-ol, and 2 beta-ol-17 beta-hemisuccinate.

Topics: Antibodies; Contraceptives, Postcoital; Esterification; Esters; Magnetic Resonance Spectroscopy; Mass Spectrometry; Norandrostanes; Radioimmunoassay

The delay in appearance of vaginal cornification associated with administration of anordiol (de-esterified anordrin) in the post-ovulatory period was confirmed. Ovarian tissue incubated in vitro for 2 h on the day which, in normal cycles, would be the day of proestrus produced negligible amounts of estradiol even in the presence of androstenedione and human menopausal gonadotropin, despite the appearance of apparently mature follicles in the ovaries. Ovaries of untreated rats produced significant amounts of estradiol when androstenedione was present. Continued incubation for 3 days resulted in significant estradiol production by ovaries of anordiol-pretreated rats in the presence of androstenedione, but less than that of ovaries of control rats. Granulosa cells of immature rats pretreated with diethylstilbestrol (DES) were unaffected by pretreatment with anordrin, whether anordrin was given before or after DES treatment. Taken together, the results indicate that anordiol inhibits development of the capacity for estrogen secretion in maturing follicles without affecting structural development, but that follicles that grow under the influence of high concentrations of estrogen (DES) are unaffected by the presence of anordrin (which is rapidly converted to anordiol in vivo). The latter result suggests that DES treatment bypasses the anordiol-sensitive step in follicular maturation.

Topics: Analysis of Variance; Androstenedione; Animals; Cells, Cultured; Contraceptives, Postcoital; Diethylstilbestrol; Drug Administration Schedule; Estrogens; Estrus; Female; In Vitro Techniques; Injections, Subcutaneous; Luteolytic Agents; Menotropins; Norandrostanes; Ovary; Radioimmunoassay; Rats; Rats, Inbred Strains

Two A-nor steroids, Anordrin and H241, showed a marked antifertility effect when given orally to hamsters at 10 mg/kg/day for three or four days after mating. Further study indicated that the antifertility effect was due to a disturbance of egg transport, retarded development and degeneration of fertilized eggs.

Topics: Animals; Cleavage Stage, Ovum; Contraceptives, Postcoital; Cricetinae; Embryo Implantation; Fallopian Tubes; Female; Norandrostanes; Ovulation; Ovum; Ovum Transport; Rabbits; Uterus