Recombinant Human Chorionic Gonadotropin Research Articles

Dual trigger of triptorelin and HCG optimizes clinical outcome for high ovarian responder in GnRH-antagonist protocols.

In this paper, a retrospective cohort study was conducted to the high ovarian responders in GnRH-antagonist protocols of IVF/ICSI cycles. The purpose of the study is to investigate whether dual triggering of final oocyte maturation with a combination of gonadotropin-releasing hormone (GnRH) agonist and human chorionic gonadotropin (HCG) can improve the clinical outcome compared with traditional dose (10000IU) HCG trigger and low-dose (8000IU) HCG trigger for high ovarian responders in GnRH-antagonist in vitro fertilization/intracytoplasmic sperm injection (IVF-ICSI) cycles. Our study included 226 couples with high ovarian responders in GnRH-antagonist protocols of IVF/ICSI cycles. Standard dosage of HCG trigger (10000 IU of recombinant HCG) versus dual trigger (0.2 mg of triptorelin and 2000 IU of recombinant HCG) and low-dose HCG trigger (8000IU of recombinant HCG) were used for final oocyte maturation. Our main outcome measures were high quality embryo rate, the number of usable embryos, the risk of OHSS, duration of hospitalization and incidence rate of complications. Our evidence demonstrated that dual trigger is capable of preventing severe OHSS while still maintaining excellent high quality embryo rate in in high ovarian responders of GnRH-antagonist protocols.

Comparison of Serum Progesterone levels and Incidence of Pregnancy with Urinary and Recombinant HCG in Women Undergoing ICSI

Objective: Luteinizing hormone (LH) surge is important in the final maturation of the oocyte and in oocyte retrieval. The structural similarity of human chorionic gonadotropin (HCG) and LH allow its binding to HCG/LH receptor. So, we aimed to compare the efficacy of urinary HCG (uHCG) and recombinant HCG (rHCG) for induction of oocyte maturation, triggering ovulation and incidence of pregnancy through assessing the serum levels of estrogen, progesterone, and βHCG in infertile women undergoing intracytoplasmic sperm injection (ICSI) cycles. Subjects and Methods: Eighty-one Egyptian females were recruited in this study including 71 infertile females undergoing ICSI and 10 apparently healthy females as a control group. The infertile female patients were classified based on the treatment used for triggering of mature follicles into 35 infertile females received 1000 IU uHCG and 36 infertile females received 500 µg rHCG. Serum levels of basal follicle stimulating hormone (FSH) and LH, estrogen (E2), progesterone (P) and βHCG were measured at different times of menstrual cycle by ELISA methods. Results: serum FSH and LH were similar in all the studied groups. Serum P level at day 5 embryo transfer and number of retrieved and mature oocytes were comparable in the rHCG and uHCG groups. While Serum P level at day oocyte pick up was significantly higher in rHCG group than in uHCG group. The successful pregnancy was higher in uHCG group than in rHCG group. Conclusion: the uHCG more effective biochemical pregnancy rate more than rHCG. However, rHCG showed equivalent efficacy to uHCG in terms of serum P level at day 5 embryo transfer, the number of retrieved oocytes and mature oocytes in female patients undergoing ICSI. While rHCG increases the serum P level at day oocyte pick up to abnormal high level which may reduce the incidence of pregnancy in females received rHCG.

Analytical characterization of recombinant hCG and comparative studies with reference product.

IntroductionRegulatory agencies recommend a stepwise approach for demonstrating biosimilarity between a proposed biosimilar and reference biological product emphasizing for functional and structural characterization to trace if there is any difference which may impact safety and efficacy. We studied the comparative structural and biological attributes of recombinant human chorionic gonadotropin (rhCG), SB005, with reference product, Ovidrel® and Ovitrelle®. Recombiant hCG was approved in 2000 by the US Food and Drug Administration for the induction of final follicular maturation, early luteinization in infertile women as part of assisted reproductive technology program. It is also indicated for the induction of ovulation and pregnancy in ovulatory infertile patients whose cause of infertility is not due to ovarian failure.Materials and methodsPrimary structure was studied by intact mass analysis, peptide fingerprinting, peptide mass fingerprinting and sequence coverage analysis. Higher order structure was studied by circular dichroism, ultraviolet-visible spectroscopy, fluorescence spectroscopy, and disulfide bridge analysis. Different isoforms of reference product and SB005 were identified using capillary isoelectric focusing and capillary zone electrophoresis. Glycosylation was studied by N-glycan mapping using LC-ESI-MS, point of glycosylation, released glycan analysis using ultra performance liquid chromatography and sialic acid analysis. Product related impurities such as oligomer content analysis and oxidized impurities were studied using size exclusion chromatography and reverse phase high performance liquid chromatography, respectively. Biological activity in term of potency of reference product and SB005 was studied by in vivo analysis.Results and ConclusionIn this study we have compared analytical similarity of recombinant rhCG (SB005) produced at Sun Pharmaceuticals with the reference product with respect to its primary, higher order structure, isoforms, charge variants, glycosylation, sialyation pattern, pharmacodynamic and in vivo efficacy. Our studies show that the in house produced rhCG has a high degree of structural and functional similarity with the reference product available in the market.

Single-dose pharmacokinetic study comparing the pharmacokinetics of recombinant human chorionic gonadotropin in healthy Japanese and Caucasian women and recombinant human chorionic gonadotropin and urinary human chorionic gonadotropin in healthy Japanese women.

PurposeRecombinant hCG (r‐hCG) was approved in Japan in 2016. As a prerequisite for a Phase III study in Japan related to this approval, the pharmacokinetic (PK) profile of r‐hCG was investigated.MethodsAn open‐label, partly randomized, single‐center, single‐dose, group‐comparison, Phase I PK‐bridging study was done that compared a single 250 μg dose of r‐hCG with a single 5000 IU dose of urinary hCG (u‐hCG) in healthy Japanese women, as well as comparing a single 250 μg dose of r‐hCG in Japanese and Caucasian women. The Japanese participants were randomized 1:1 to receive either r‐hCG or u‐hCG, while the Caucasian participants were weight‐matched to the Japanese participants who were receiving r‐hCG in a 1:1 fashion. The primary PK parameters were the area under the serum concentration–time curve from time 0 extrapolated to infinity (AUC 0–∞) and the maximum serum concentration (Cmax).ResultsThe mean serum hCG concentration–time profiles of r‐hCG in the Japanese and Caucasian participants were a similar shape, but the level of overall exposure was ~20% lower in the Japanese participants. For the Japanese participants, r‐hCG resulted in an 11% lower Cmax but a 19% higher AUC 0–∞ compared with u‐hCG. No new safety signal was identified.ConclusionThis study cannot exclude a potential difference in the PK profile of r‐hCG between Japanese and Caucasian participants. However, this study does not indicate that there are clinically relevant differences in the serum PK of r‐hCG and u‐hCG in the Japanese participants.

Efficacy of patient self-administered recombinant human chorionic gonadotropin (RHCG) is comparable to nurse administred RHCG in ovulation induction cycles

Infertility treatment requires patients to overcome a variety of physical, emotional, and financial obstacles. Often, patients express anxiety over self-administration of medications, including subcutaneous recombinant hCG (rHCG). Midluteal rise in progesterone (P4) levels to >3ng/dL suggest evidence of ovulation, and of ≥9ng/dL of adequate luteinization. Many patients express concern over whether they will adequately be able to self-administer medication. The study sought to determine if the rate and adequacy of ovulatory response is comparable whether a patient or a nurse administers the medication.

Central hypogonadism due to a giant, “silent” FSH-secreting, atypical pituitary adenoma: effects of adenoma dissection and short-term Leydig cell stimulation by luteinizing hormone (LH) and human chorionic gonadotropin (hCG)

We present a case report of an atypical giant pituitary adenoma secreting follicle-stimulating hormone (FSH). A 55-year-old patient presented for erectile dysfunction, loss of libido and fatigue. The biochemical evaluation showed very high FSH serum levels in the presence of central hypogonadism. Neither testicular enlargement nor increased sperm count was observed, thus a secretion of FSH with reduced biological activity was supposed. The histological examination after neuro-surgery showed an atypical pituitary adenoma with FSH-positive cells. Hypogonadism persisted and semen analyses impaired until azoospermia in conjunction with the reduction in FSH levels suggesting that, at least in part, this gonadotropin should be biologically active. Thus, we hypothesized a concomitant primary testicular insufficiency. The patient underwent short-term treatment trials with low doses of either recombinant luteinizing hormone (LH) or human chorionic gonadotropin (hCG) in three consecutive treatment schemes, showing an equal efficacy in stimulating testosterone (T) increase. This is the first case of atypical, giant FSH-secreting pituitary adenoma with high FSH serum levels without signs of testicular hyperstimulation, in presence of hypogonadism with plausible combined primary and secondary etiology. Hypophysectomized patients may represent a good model to assess both pharmacodynamics and effective dose of LH and hCG in the male.

Superovulatory responses in cynomolgus monkeys (Macaca fascicularis) depend on the interaction between donor status and superovulation method used.

The current study was performed to investigate the effect of oocyte donor status, including age and body weight, on metaphase II (MII) oocyte recovery using two superovulation methods in cynomolgus monkeys. The use of Method A [recombinant gonadotrophin (75 IU/kg, 3 ×, 3-day intervals) and human chorionic gonadotropin (hCG)] led to great increases in ovary size and the mean number of MII oocytes retrieved in age- and body-weight-dependent manner; in contrast, both the parameters were similar in Method B [recombinant gonadotrophin (60 IU, twice daily, 6 days), recombinant gonadotropin and recombinant human luteinizing hormone (rhLH) (60 IU, twice daily, 3 days), and hCG]. Importantly, Method A showed maximal MII oocyte recovery rate in > 60-month-old or 4.5–5.0-kg female monkeys, whereas Method B was equally effective regardless of the donor age and body weight. These results indicate that superovulatory responses depend on the interaction between oocyte donor status and the superovulation method used in cynomolgus monkeys.

Gonadal response after a single-dose stimulation test with recombinant human chorionic gonadotropin (rhCG) in patients with isolated prepubertal cryptorchidism.

BackgroundThe evaluation of prepubertal gonadal Leydig cells secretion requires gonadotropin stimulation. Urinary hCG (human chorionic gonadotropin) is currently unavailable in many countries, however, recombinant hCG (rhCG) can be used. Our aim was to evaluate rhCG-stimulated testicular hormones in a group of patients with cryptorchidism.MethodsWe evaluated 31 prepubertal boys (age range, 0.75–9.0 years) presenting with unilateral (n = 24) or bilateral (n = 7) cryptorchidism. Patients with other genital abnormalities, previous use of hCG or testosterone or previous surgeries were excluded. Blood samples were obtained at baseline and 7 days after a single subcutaneous dose of rhCG (Ovidrel® 250 mcg) to measure the testosterone, DHT (dihydrotestosterone), AMH (anti-Mullerian hormone), and inhibin B levels.ResultsrhCG stimulation significantly increased testosterone levels from 10 ng/dl to 247.8 ± 135.8 ng/dl, increased DHT levels from 4.6 ± 0.8 to 32.3 ± 18.0 ng/dl, and increased the T/DHT ratio from 2.2 ± 0.4 to 8.0 ± 3.5. There was also a significant increase in inhibin B (from 105.8 ± 65.2 to 132.4 ± 56.1 pg/ml; p < 0.05) and AMH levels (from 109.4 ± 52.6 to 152.9 ± 65.2 ng/ml; p < 0.01) after the rhCG stimulation.ConclusionsIn this cohort, hormonal responses can be elicited after the rhCG stimulation test, suggesting that rhCG is a promising stimulation test to replace the urinary hCG test during the evaluation of gonadal Leydig cells function. The clinical applicability and adequate performance of rhCG testing must be investigated in future studies.Electronic supplementary materialThe online version of this article (doi:10.1186/s12610-016-0039-2) contains supplementary material, which is available to authorized users.

Dual trigger with gonadotropin-releasing hormone agonist and recombinant human chorionic gonadotropin improves in vitro fertilization outcome in gonadotropin-releasing hormone antagonist cycles.

The aim of this study was to evaluate whether dual trigger with leuprolide acetate plus recombinant human chorionic gonadotropin (hCG) improves in vitro fertilization outcome in gonadotropin-releasing hormone antagonist cycles. A total of 156 patients diagnosed with mild male factor, unexplained or tubal factor infertility were enrolled in the study. All subjects were allocated into one of two groups: the dual trigger group (leuprolide acetate 500μg + recombinant hCG 250μg) and the standard group (recombinant hCG 250μg) according to the selected trigger method. Oocyte trigger was performed when at least three follicles >17 mm were observed. Pregnancy rate, number of collected oocytes, number of metaphase II oocytes, number of grade-A embryos, cycle cancellation rate, and ovarian hyperstimulation syndrome rate were the main outcome measures for the study. The mean number of grade-A embryos (1.6 ± 1.5 vs 1.1 ± 1.4, P=0.01) and of metaphase II oocytes (7.9 ± 4.6 vs 6.3 ± 5.8, P=0.02) was significantly higher in the dual-trigger group. Pregnancy rate was significantly higher in the dual-trigger group than in the standard group (54.8 vs 37.5%, P=0.006). Two cases of mild ovarian hyperstimulation syndrome were observed in each group. This novel and more physiological trigger approach using 500μg leuprolide acetate plus 250μg recombinant hCG may lead to an increase in the number of metaphase II oocytes, grade-A embryos, and may improve pregnancy rates.

Recombinant versus urinary human chorionic gonadotrophin for final oocyte maturation triggering in IVF and ICSI cycles.

For the last few decades urinary human chorionic gonadotrophin (uhCG) has been used to trigger final oocyte maturation in cycles of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Recombinant technology has allowed the production of two drugs, recombinant human chorionic gonadotrophin (rhCG) and recombinant luteinising hormone (rLH), that can be used for the same purpose, to mimic the endogenous luteinising hormone (LH) surge. This allows commercial manufacturers to adjust production according to market requirements and to remove all urinary contaminants, facilitating the safe subcutaneous administration of a compound with less batch-to-batch variation. However, prior to a change in practice, it is necessary to compare the effectiveness of the recombinant drugs to the currently used urinary human chorionic gonadotrophin (uhCG). To assess the effects of subcutaneous rhCG and high dose rLH versus uhCG for inducing final oocyte maturation in subfertile women undergoing IVF and ICSI cycles. We searched the Cochrane Menstrual Disorders and Subfertility Group Trials Register (April 2015), the Cochrane Central Register of Controlled Trials (CENTRAL) (2015, Issue 3), MEDLINE (1946 to April 2015), EMBASE (1980 to April 2015) and PsycINFO (1806 to April 2015) as well as trial registers at ClinicalTrials.gov on 13 May 2015 and the World Health Organization International Clinical Trials Registry Platform (WHO ICTRP) search portal on 14 May 2015. Two review authors independently scanned titles and abstracts and selected those that appeared relevant for collection of the full paper. We included randomised controlled trials comparing rhCG and rLH with urinary hCG for final oocyte maturation triggering in IVF and ICSI cycles for treatment of infertility in normogonadotropic women. Two authors independently performed assessment for inclusion or exclusion, quality assessment and data extraction. We discussed any discrepancies in the presence of a third author to reach a consensus. The primary review outcomes were ongoing pregnancy/live birth and incidence of ovarian hyperstimulation syndrome (OHSS). Clinical pregnancy, miscarriage rate, number of oocytes retrieved and adverse events were secondary outcomes. We combined data to calculate pooled odds ratios (ORs) and 95% confidence intervals (CIs) and assessed statistical heterogeneity using the I(2) statistic. We evaluated the overall quality of the evidence for the main comparisons using GRADE methods. We included 18 RCTs involving 2952 participants; 15 compared rhCG with uhCG, and 3 compared rhLH with uhCG. The evidence for different comparisons ranged from very low to high quality: limitations were poor reporting of study methods and imprecision. Pharmaceutical companies funded 9 of the 18 studies, and 5 studies did not clearly report funding source. Ongoing pregnancy/live birthThere was no conclusive evidence of a difference between rhCG and uhCG (OR 1.15, 95% CI 0.89 to 1.49; 7 RCTs, N = 1136, I(2) = 0%, moderate quality evidence) or between rhLH and uhCG (OR 0.95, 95% CI 0.51 to 1.78, 2 RCTs, N = 289, I(2) = 0%, very low quality evidence) for ongoing pregnancy/live birth rates. OHSS There was no evidence of a difference between rhCG and uhCG in the incidence of OHSS: moderate to severe OHSS (OR 1.76, 95% CI 0.37 to 8.45; 3 RCTs, N = 417, I(2) = 0%, low quality evidence), moderate OHSS (OR 0.78, 95% CI 0.27 to 2.27; 1 RCT, N = 243, I(2) = 0%, low quality evidence), mild to moderate OHSS (OR 1.00, 95% CI 0.42 to 2.38; 2 RCTs, N = 320, I(2) = 0%, low quality evidence) or undefined OHSS (OR 1.18, 95% CI 0.50 to 2.78; 3 RCTs, N = 495, I(2) = 0%, low quality evidence). Likewise, there was no evidence of a difference between rhLH and uhCG in OHSS rates for moderate OHSS (OR 0.82, 95% CI 0.39 to 1.69, 2 RCTs, N = 280, I(2) = 5%, very low quality evidence). Other adverse events There was no evidence of a difference in miscarriage rates between rhCG and uhCG (OR 0.72, 95% CI 0.41 to 1.25; 8 RCTs, N = 1196, I(2) = 0%, low quality evidence) or between rhLH and uhCG (OR 0.95, 95% CI 0.38 to 2.40; 2 RCTs, N = 289, I(2) = 0%, very low quality evidence). For other adverse effects (most commonly injection-site reactions) rhCG was associated with a lower number of adverse events than uhCG (OR 0.52, 95% CI 0.35 to 0.76; 5 RCTS, N = 561; I(2) = 67%, moderate quality evidence). However, when we used a random-effects model due to substantial statistical heterogeneity, there was no evidence of a difference between the groups (OR 0.56, 95% CI 0.27 to 1.13). Only one study comparing rLH and uhCG reported other adverse events, and it was impossible to draw conclusions. We conclude that there is no evidence of a difference between rhCG or rhLH and uhCG for live birth or ongoing pregnancy rates or rates of OHSS.

Salvage hormonal therapy after failed microdissection testicular sperm extraction: A multi-institutional prospective study.

To validate the efficacy of salvage hormonal therapy in men with non-obstructive azoospermia at their second microdissection testicular sperm extraction. This was a multi-institutional study registered at the Japanese University Hospital Medical Information Network clinical trial center. After 1month of human chorionic gonadotropin therapy (5000IU, three times a week), patients were treated with recombinant human follicle-stimulating hormone (150IU, three times a week) and human chorionic gonadotropin for the next 3months. Three testicular samples were obtained randomly from both testes, and sent for pathological diagnosis at the first and second microdissection testicular sperm extraction. A total of 21 men, excluding those with chromosomal abnormalities, azoospermia factora or b deletions, extremely small testes (<2mL), or prior hormonal therapy, were eligible to participate based on our inclusion criteria. At the first microdissection testicular sperm extraction, 13 and six patients had Sertoli cells only and an early maturation arrest, respectively. With the second microdissection testicular sperm extraction, sperm were successfully obtained from two patients (10%). Patient age, testicular volume and hormone profiles were not associated with the results of the second microdissection testicular sperm extraction. However, the testicular histology of the two successful patients were late maturation arrest and hypospermatogenesis. Effectiveness of human chorionic gonadotropin-based salvage hormonal therapy preceding a second microdissection testicular sperm extraction seems to be limited. Non-obstructive azoospermia men who have differentiated cells in their testes are likely to respond to hormonal stimulation.

A Preliminary Report of A Low-Dose Step-Up Regimen of Recombinant Human FSH for Young Women Undergoing Ovulation Induction with IUI.

The aim of this study was to evaluate the efficacy and safety of a recombinant human follicle stimulating hormone (r-FSH) low-dose step-up regimen for controlled ovarian hyperstimulation in patients undergoing ovulation induction (OI) with intrauterine insemination (IUI). The study was conducted in the Department of Obstetrics and Gynecology, Far Eastern Memorial Hospital, New Taipei, Taiwan. In this prospective, observational study, consecutive infertile women (20-35 years) with regular menstrual cycles and a normal baseline FSH level were prospectively enrolled between January 2010 and September 2010. A starting dose of 112.5 IU/day r-FSH was administered on day 3 and increased by 37.5 IU/day every 2 days until a follicle ≥11 mm in diameter was present. Recombinant human chorionic gonadotropin (r-hCG) was administered when a follicle ≥18 mm was noted. Monifollicular development was defined as only one follicle with a diameter ≥16 mm. Clinical pregnancy was defined as a pregnancy diagnosed by ultrasonographic visualization of one or more gestational sacs. A total of 29 women and 30 cycles were included. The mean daily dose of r-FSH to achieve a follicle of ≥11 mm in diameter was 131.3 ± 23.6 IU and the mean total dose was 1030.0 ± 383.2 IU. Approximately 41% of the cycles were monofollicular. Clinical pregnancy was observed in 9 (30.0%) cycles, and a fetal heart beat was observed in 7 (23.3%). There were no multiple pregnancies. Mild ovarian hyperstimulation syndrome, which was resolved with conservative management, was observed in 3 (10.0%) cycles. This r-FSH low-dose step-up regimen seems to be a feasible and practical method for OI in younger infertile women undergoing IUI.

Impact of quantitative parameters, such as number of retrieved oocytes, number of transferred embryos and availability of surplus embryos for cryopreservation, on clinical pregnancy rates in ART

Objective: To estimate the impact of quantitative parameters such as number of retrieved oocytes, number of transferred embryos and availability of surplus embryos for cryopreservation on clinical pregnancies in assisted reproductive technology (ART).Design: We used the database of fertility clinic on in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment cycles from year 2000 to 2010. Follicular fluids containing oocyte–cumulus complexes were recovered by a single lumen follicle aspiration needle, and IVF or ICSI procedure was used depending on infertility indications. Fertilization determination was performed on the second day. Zygotes were transferred into fresh medium and cultivated until the embryo transfer procedure. According to morphological criteria, the best quality embryos were transferred into uterus on the second or third day. Surplus embryos were frozen by slow freezing or by vitrification. We evaluated only clinical pregnancies that were diagnosed by ultrasonography.Patients: We allocated all 1161 women cohort into three age groups: 671 women were <35; 397 were 35–40 years old and 93 women were >40. Indications for IVF treatment were as follows: mechanical factor (436 cases), male infertility (361 cases), idiopathic infertility (129 cases), endometriosis (78 cases), immunological infertility (14 cases), anovulation (28 cases), and other indications (28 cases). There were no data about the cause of infertility of 87 patients. The stimulation protocol was composed from gonadotropin hormone stimulation with antagonist or agonist supplementation. The triggering of luteinizing hormone (LH) surge was performed by recombinant human chorionic gonadotropin (hCG). The clinical pregnancy rates were: 42.2% (283/671) for women <35, 31.2% (124/397) for women 35–40-year-old and 16.1% (15/93) for women >40.Interventions: All materials and methods that we used were based on results from our daily practice in IVF clinic and have no experimental design.Main outcome measures: We evaluated clinical pregnancies in terms of the number of obtained oocytes, the amount of transferred embryos and the availability of surplus embryos for freezing. The number of retrieved oocytes was divided as follows: 1–5 (group I); 6–10 (group II); ≥11 (group III). The number of transferred embryos was from 1 to 3. Cases at which cryopreservation of embryos was not performed were evaluated as well.Results: For women <35 and 35–40 years old, the highest percentage of pregnancies was achieved when 11 and more oocytes were obtained. Differences in women under 35 between groups were statistically significant. Statistically significant differences in clinical pregnancies depend on the number of transferred embryos in the age groups of <35 and 35–40. Women <35 and 35–40 years old, with the availability of surplus embryos for freezing, had better chances for pregnancy compared with women, who had no excess embryos for freezing.Conclusions: The number of obtained oocytes, the number of transferred embryos and the availability of surplus embryos for freezing may serve as predictors of pregnancy rates in ART.

Atrazine-Mediated Disruption of Steroidogenesis in BLTK1 Murine Leydig Cells.

Atrazine (ATR) is a broad-spectrum triazine herbicide that disrupts steroidogenesis resulting in reproductive and developmental toxicity at high doses. Mouse BLTK1 Leydig cells were used as a steroidogenic model to investigate the effects of ATR on testosterone (T) biosynthesis. Induction of steroidogenesis by 3 ng/ml recombinant human chorionic gonadotropin (rhCG) induced intracellular 3',5' cyclic adenosine monophosphate (cAMP) approximately 20-fold and T approximately 3-fold at 4 h. Co-treatment with 300 μM ATR super-induced cAMP levels 100-fold yet antagonized rhCG-mediated induction of T approximately 20% at 4 h. ATR inhibited cAMP-specific phosphodiesterase (cPDE) with an IC50 of ≥98 μM, suggesting cPDE inhibition contributes to the super-induction of cAMP. However, concentrations of up to 3 mM db-cAMP did not antagonize rhCG induction of T levels, suggesting cAMP super-induction alone does not decrease T biosynthesis. Western analysis of cAMP-activated protein kinase A (PKA) target proteins identified ATR-mediated concentration-dependent alterations in phosphorylation including phospho-CREB. These results suggest the cPDE inhibition by ATR and super-induction of cAMP are independent of effects on T levels, and that altered phosphorylation of key steroidogenic regulatory proteins may underlie ATR-mediated disruption of steroidogenesis.

Live birth following early follicular phase oocyte collection and vitrified-warmed embryo transfer 8 days later

A 30-year-old woman with premature ovarian insufficiency had two follicles measuring 17 mm and 14 mm on day 3 of her menstrual cycle. Serum oestradiol concentration was 210 pg/ml. Recombinant human chorionic gonadotrophin was given and 5 mg/day letrozole started orally. One metaphase II oocyte was collected 36 h later. A 4-cell embryo was vitrified on the second day after fertilization. Letrozole was stopped on cycle day 8 due to absence of any other visible antral follicles. Oestradiol valerate 6 mg/day was started and the endometrium was 9.2 mm on cycle day 11. The embryo was warmed and transferred on cycle day 13, the 8th day after oocyte retrieval. Luteal phase support with progesterone, oestradiol and low molecular weight heparin was started on the day of transfer and continued until the 10th gestational week. A healthy girl weighing 3200 g was born at term. Early follicular phase oocyte collection did not result in early opening of the implantation window. Apparently secretory transformation was not started until luteal phase support, enabling a cleavage stage embryo transferred 8 days later to implant. Either corpus luteum formation could be disrupted or the endometrium could remain unresponsive to progesterone during the early follicular phase.

Corifollitropin alfa combined with GNRH agonist triggering appears to be safe and efficient option in oocyte donation cycles. A prospective longitudinal study

To explore whether differences existed between the use of (Elonva®) and gonadotrophin releasing hormone (GnRH) agonist for triggering and the traditional stimulation schema with recombinant FSH and human chorionic gonadotrophin (hCG) for triggering in terms of efficacy, safety and patient friendliness in oocytes donation cycles.

Pharmacokinetics and Safety Profile of DA-3803, a Proposed Biosimilar of Recombinant Human Chorionic Gonadotropin, in Healthy Subjects.

DA-3803 is a proposed biosimilar of recombinant human chorionic gonadotropin (r-hCG), which is a therapeutic protein used to treat infertility. We compared the pharmacokinetics and safety profiles of DA-3803 (test) with those of a conventional formulation (Ovidrel(®), reference). A single-blind, randomized, single-dose, two-period, two-sequence, crossover study was conducted in 32 healthy subjects. In each period, 250 μg of r-hCG of the test or reference formulation was administered subcutaneously with a 3-week washout period. Serial blood samples were obtained for pharmacokinetic analysis and blood levels of human chorionic gonadotropin (hCG) were determined. The geometric mean ratios (GMRs) with 90 % confidence intervals (CIs) for the maximum (peak) plasma concentration (C max) and the area under the plasma concentration-time curve (AUC) were estimated. Among the 32 subjects, 27 completed the study. No serious adverse events were observed. The mean concentration-time profiles of the test formulation tended to be higher than those of the reference formulation. The mean C max values of the two products were similar (142 mIU/mL for reference vs. 143 mIU/mL for test), but the mean AUC from time zero to infinity (AUC∞) values of the test drug were approximately 25 % higher than those of the reference drug (9403 mIU·h/mL for reference vs. 11,817 mIU·h/mL for test). The GMR (90 % CI) of the test formulation to the reference formulation for C max, AUC from time zero to the last measurable time (AUClast), and AUC∞ were 1.03 (0.93-1.14), 1.25 (1.18-1.33), and 1.27 (1.19-1.35), respectively. A single subcutaneous injection of either DA-3803 or Ovidrel(®) was safe and well-tolerated. A comparative pharmacokinetics study showed that DA-3803 was bioequivalent to Ovidrel(®) in the C max value but non-equivalent in the AUC value. These pharmacokinetic differences may not be expected to affect therapeutic efficacy of these two drugs, but further clinical studies are warranted to confirm their therapeutic equivalence.

Lack of Serum anti-Mullerian hormone responses after recombinant human chorionic gonadotropin stimulation in women with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is an anovulatory disorder characterized by excess androgen production and increased LH secretion. Serum anti-Mullerian hormone (AMH) is also elevated in this disorder. Women with PCOS exhibit a positive correlation between AMH and LH levels and recent in vitro data demonstrate that LH can directly stimulate AMH production by granulosa cells from women with PCOS. The objective of the study was to directly test whether LH increases AMH production in women with PCOS in vivo by assessing responses after recombinant human chorionic gonadotropin (r-hCG) stimulation. This was a prospective study. The study was conducted at a research center at an academic medical center. Women with PCOS (n = 28) and normal controls (n = 29) participated in the study. Blood samples were obtained before and 24 hours after iv administration of 25 μg r-hCG. Basal and stimulated serum AMH, androstenedione, T, and 17-hydroxyprogesterone levels were measured. Baseline AMH levels in women with PCOS were greater than in normal controls and correlated with levels of LH as well as androstenedione, T, and 17-hydroxyprogesterone. A rise of serum AMH levels was not observed after r-hCG administration in women with PCOS or normal ovaries. These findings are in contrast to in vitro evidence demonstrating that AMH secretion by granulosa cells of PCOS women in response to LH stimulation and suggest AMH regulation in vivo is complex and that the elevated serum AMH in women with PCOS is not a direct effect of the excess LH production characteristic of PCOS.

Limitations in the process of transcription and translation inhibit recombinant human chorionic gonadotropin expression in CHO cells

Human chorionic gonadotropin (hCG) is a glycoprotein hormone that exists as a heterodimer with a α subunit and β subunit assembled together with disulfide bridges. This hormone plays an important role in the detection of ovulation induction and in the treatment of certain diseases that cause female infertility. The effects of transcription, subunit expression, assembling and secretion on recombinant hCG expression in CHO cells were studied using stable high-producing and low-producing cell lines generated by the FLP-In™ system. The results indicated that the mRNA and polypeptide levels of the β subunit were always higher than those of the α subunit. Further study confirmed that the differences were caused by the transcription rate rather than by mRNA stability. In the high-producing cell lines, there was obvious transcription level limitation of the α subunit in contrast to the β subunit. In addition, there was obvious limitation of the synthetic steps from mRNA to polypeptide for both the α subunit and the β subunit, especially the β subunit. Significant limitations of the assembly and secretion levels were not observed in this research. This study presents a research methodology for double subunit protein expression and provides valuable evidence for the enhancement of recombinant hCG productivity.

Single-arm, observational study of the ease of use of a redesigned pen device to deliver recombinant human follicle-stimulating hormone (follitropin alfa) for assisted reproductive technology treatment.

PurposeEvaluation of patients’ ease of use of the redesigned, disposable, ready-to-use follitropin alfa pen during controlled ovarian stimulation for assisted reproductive technology.MethodsThis single-center, observational, open-label, single-arm study recruited infertile normo-ovulatory women (aged 18–45 years). Nurses trained patients to self-administer recombinant human follicle-stimulating hormone daily using the follitropin alfa pen (300 IU, 450 IU, and 900 IU). Before treatment, patients completed Questionnaire A. Following self-administered treatment, on stimulation days 5–6 and 7–8 (within a day of receiving recombinant human chorionic gonadotropin), patients completed Questionnaire B. Nurses completed an ease-of-learning/teaching questionnaire. The primary endpoint was proportion of patients rating the pen as “easy/very easy” to use (Questionnaire B) on the final visit before recombinant human chorionic gonadotropin. Secondary endpoints included: proportion of patients rating the follitropin alfa pen as easy to learn, use, prepare, deliver, and dispose of (Questionnaires A and B). Proportions (95% confidence intervals [CIs]) were provided for primary and secondary endpoints. Adverse events were reported descriptively.ResultsEighty-six patients received recombinant human follicle-stimulating hormone. Of the 72 patients who had completed the overall assessment questions, 66 (91.7%; 95% CI =82.7%–96.9%) found the pen “easy” to use. Also, 70/86 (81.4%) patients “strongly agreed/agreed” that, overall, it was easy to learn how to use the pen; 72/86 (83.7%) “strongly agreed/agreed” that easily understandable, verbal information was provided; and 70/86 (81.4%) were confident about using the pen correctly. In total, 24/26 nurses (92.3%; 95% CI =74.9%–99.1%) rated the pen as easy to use. Clinical pregnancy rate/patient/cycle/embryo transfer was 37%. Twenty-six ovarian hyperstimulation syndrome events were reported (none severe; 16 patients [19%]); of these, 13 occurred at embryo transfer.ConclusionIn this observational study, patients had a high acceptance of the redesigned follitropin alfa pen, with most finding it very easy/easy to use. Assisted reproductive technology nurses found the pen very easy/easy to teach.