GnRH-I receptor (GnRHR) [1][2]

In human ovarian cancer OV-1063 xenografts in nude mice, chronic treatment with Cetrorelix significantly inhibited tumor growth, whereas the GnRH-I agonist Triptorelin did not [1].
In 20 IVF patients with ovarian stimulation, Cetrorelix administered subcutaneously at 3 mg/day (n=15) or 1 mg/day (n=5) from day 7 of the menstrual cycle until HCG injection effectively suppressed endogenous LH surges, with no patient experiencing a premature LH peak. Mean oocytes collected were 8.1 per patient, fertilization rate 61.5%, and three pregnancies were achieved [2].
In Balb/c mice, Cetrorelix (0.5 mg/kg/day s.c. for 16 days) significantly reduced cyclophosphamide-induced ovarian follicle destruction. At cyclophosphamide 50 mg/kg, Cetrorelix pretreatment resulted in only 14% loss of primordial follicles vs. 53% in control (P<0.001); at 75 mg/kg, 35% loss vs. 54% (P<0.004). Relative protection ratios were 1.83 and 1.4, respectively [3].

Female Balb/c mice (8-9 weeks old, weight 15-25 g) were injected subcutaneously daily with Cetrorelix at 0.5 mg/kg from day 1 to day 15. On day 9, a single intraperitoneal dose of cyclophosphamide (50 or 75 mg/kg) was administered. Control groups received saline. On day 16, mice were sacrificed, both ovaries removed, fixed in 4% paraformaldehyde, embedded in paraplast, serially sectioned at 5 μm, stained with hematoxylin-eosin, and primordial follicles (single layer of squamous pregranulosa cells, no theca layer, nucleus clearly identified) were counted in every tenth section; total number per mouse was calculated by multiplying by 10 [3].

Absorption, Distribution and Excretion
Absorption is rapid following subcutaneous injection. The mean absolute bioavailability after subcutaneous administration in healthy female subjects was 85%. Unmetabolized cetrarolix was detected in urine after subcutaneous injection of 10 mg cetrarolix in both men and women. Dosage: 1.16 L/kg Dosage: 1.28 ml/min·kg [Single subcutaneous injection of 3 mg in healthy adult women] Unmetabolized cetrarolix was detected in urine after subcutaneous injection of 10 mg cetrarolix in both men and women. Cetrolix and trace amounts of (1-9), (1-7), (1-6), and (1-4) peptides were detected in bile samples 24 hours later. 2-4% of the dose was excreted unchanged in the urine as cetrarolix, and 5-10% of the dose was excreted in the bile as cetrarolix and its four metabolites. Therefore, only 7-14% of the total dose is recovered in unchanged cetrorelix and its metabolites from urine and bile within 24 hours. Due to the short collection time of bile and urine, the remaining dose may not be recovered. Following a single intravenous injection of 3 mg cetrorelix, its volume of distribution is approximately 1 L/kg. In vitro human plasma protein binding is 86%. In patients undergoing controlled ovarian stimulation, cetrorelix concentrations in follicular fluid and plasma are similar on the day of oocyte retrieval. Following subcutaneous injections of 0.25 mg and 3 mg cetrorelix, plasma cetrorelix concentrations are below or within the lower limit of quantitation on the days of oocyte retrieval and embryo transfer. Cetrorelix is rapidly absorbed after subcutaneous injection, reaching maximum plasma concentrations approximately 1-2 hours after administration. The mean absolute bioavailability of cetrorelix after subcutaneous injection in healthy female subjects is 85%. Pharmacokinetic studies were primarily conducted in rats and dogs. Absorption at the subcutaneous injection site is rapid and complete, regardless of sex or species. Dose-plasma AUC is linear. Cetrorelix distributes rapidly. Its primary target organs are the kidneys, liver, small intestine, and organs containing luteinizing hormone-releasing hormone (LHRH) receptors (pituitary gland, ovaries). Plasma protein binding is 86%. The drug is rapidly cleared from most tissues, primarily within 48 hours. …Cetrolix crosses the placenta in small amounts. The distribution of cetrorelix or its metabolites in breast milk has not been studied. Cetrorelix is excreted unchanged in the urine and metabolized by peptidases in the bile. …Studies in healthy volunteers indicate that cetrorelix is excreted similarly in humans, rats, and dogs. Following subcutaneous injection, the absolute bioavailability of cetrorelix is approximately 85% in both men and women. The apparent volume of distribution is 1.16 ± 0.29 L/kg in women and 1.02 ± 0.33 L/kg in men. The terminal half-life is approximately 10 hours after intravenous injection and approximately 30 hours after subcutaneous injection, with a decreasing trend in women. Human plasma protein binding is approximately 85%. Linear pharmacokinetics were observed after single (0.25, 0.5, and 1.00 mg) and multiple (0.25 to 1.00 mg) doses. Pharmacokinetics were linear within the 3 mg dose range.
Metabolism/Metabolites
In vitro studies showed that cetrorex was stable for both phase I and phase II metabolism. Cetrorex can be converted by peptidases, with peptides (1–4) being the major metabolites.
In rat bile, the major metabolite of cetrorex was identified as heptapeptide (1–7). This metabolite had no pharmacological activity in rats, i.e., it did not inhibit testosterone secretion.
After subcutaneous injection of 10 mg cetrorex into male and female rats, cetrorex and trace amounts of peptides (1–9), (1–7), (1–6), and (1–4) were detected in bile samples within 24 hours. In vitro studies showed that cetrorex was stable for both phase I and phase II metabolism. Cetrotide is converted by peptidase, with (1-4) peptides being the main metabolites.

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Biological half-life
~62.8 hours
In humans, the terminal half-lives after intravenous and subcutaneous injection are 8-9 hours and 24-40 hours, respectively.
In rats, the terminal half-lives after intravenous and subcutaneous injection are 1-2 hours and 7-14 hours, respectively… Elimination half-life: Single 3 mg dose: 62.8 hours (38.2–108 hours); Single 0.25 mg dose: 5.0 hours (2.4–48.8 hours); 0.25 mg daily for 14 days: 20.6 hours (4.1–179.3 hours) /Excerpt from table/
Half-lives greater than or equal to 100 hours were observed in excretory organs (liver, kidney), spleen, and organs containing LHRH binding sites.

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Cetrorelix causes immediate suppression of gonadotropins (LH more than FSH) within a few hours after subcutaneous injection; effects are fully reversible and dose-dependent [2].

Protein binding 86% hr Non-human toxicity value 68.1 mg/kg was determined as the minimum lethal dose.

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Cetrorelix showed no systemic toxicity and no teratogenic potential in clinical studies. Occasionally, short-term erythema at the injection site was observed, but no wheal or pruritus [2].

[1]. Reprod Biol Endocrinol.2003 Oct 7;1:65;

[2]. Suppression of the endogenous luteinizing hormone surge by the gonadotrophin-releasing hormone antagonist Cetrorelix during ovarian stimulation. Hum Reprod. 1994 May;9(5):788-91.

[3]. The GnRH antagonist cetrorelix reduces cyclophosphamide-induced ovarian follicular destruction in mice. Hum Reprod. 2004 Jun;19(6):1294-9.

Therapeutic Uses
Cetrorelix is indicated for the suppression of premature luteinizing hormone (LH) surges in women undergoing controlled ovarian stimulation. This randomized, placebo-controlled, single-blind study was conducted in 45 adult female Wistar rats… After peritoneal implantation of endometrial tissue, rats were randomly assigned to three equal intervention groups: (i) a control group, (ii) a leuprorelin group, and (iii) a cetrorelix group. Six weeks later, the implant volume was measured (volume-1) via a second laparotomy. Subsequently, the control group received weekly subcutaneous injections of saline (0.1 mL/rat), the leuprorelin group received twice-daily subcutaneous injections of leuprorelin (0.075 mg/kg), and the cetrorelix group received subcutaneous injections of cetrorelix (0.001 mg/rat/day) for 8 weeks. At the end of treatment, the implant volume was measured again (volume-2) via a third laparotomy, and the implant was completely removed for histopathological examination. Compare the volume-1 and volume-2 values within each group, and the intergroup scores for stromal and glandular tissue. In both the leuprorelin and cetrorelix groups, volume-2 was significantly reduced compared to volume-1 (P < 0.01 and P < 0.01, respectively), while there was no significant change in volume in the control group (P > 0.05). Compared to the control group, both glandular and stromal tissue were significantly reduced in the control group (P < 0.01 and P < 0.01, respectively). Leuprorelin and cetrorelix showed similar efficacy in reducing the size and histological structure of experimental endometriosis lesions. Drug Warnings: Cetrorelix should be prescribed by a healthcare professional experienced in fertility treatment. Pregnancy must be ruled out before initiating cetrorelix acetate treatment.
In patients receiving controlled ovarian stimulation, 1-2% reported elevated liver function test results, including ALT (SGPT), AST (SGOT), gamma-glutamyl transferase (GGT, GGTP), and alkaline phosphatase, up to 3 times the upper limit of normal.
Caution should be exercised in patients with GnRH allergy. These patients should be closely monitored after the first injection. In a study of an indication unrelated to infertility, one patient experienced a severe allergic reaction with cough, rash, and hypotension after 7 months of treatment with cetrorelix 10 mg/day.
Local reactions (e.g., redness, erythema, ecchymosis, pruritus, swelling, and itching) have been reported. These adverse reactions are usually transient, mild, and short-lived.
For more complete data on drug warnings for cetrorelix (8 warnings total), please visit the HSDB record page.

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Pharmacodynamics
Citrolix is a synthetic decapeptide with gonadotropin-releasing hormone (GnRH) antagonistic activity. GnRH induces anterior pituitary gonadotropic cells to produce and release luteinizing hormone (LH) and follicle-stimulating hormone (FSH). GnRH release is enhanced due to positive feedback from estradiol (E2) in the mid-menstrual cycle, leading to an LH surge. This LH surge induces ovulation of the dominant follicle, resumption of meiosis in the oocyte, and subsequent luteinization, manifested as elevated progesterone levels. Cetrorelix competes with natural GnRH for binding to pituitary cell membrane receptors, thereby controlling LH and FSH release in a dose-dependent manner.

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Cetrorelix is a GnRH antagonist that competitively inhibits GnRH receptors, suppressing LH and FSH secretion. It has been used in IVF to prevent premature LH surges [2] and in animal studies to protect ovaries from cyclophosphamide-induced damage [3]. In human ovarian cancer cells, Cetrorelix exhibits direct antiproliferative effects comparable to GnRH-I agonists in most cell lines (except EFO-27), and these effects persist after GnRH-I receptor knockdown, suggesting they are not mediated through the GnRH-I receptor [1]. In ES-2 ovarian cancer cells, Cetrorelix inhibited growth only at 1000 ng/ml [1].

C70H92CLN17O14

1431.061

1429.669

120287-85-6

Cetrorelix Acetate;145672-81-7;Cetrorelix diacetate;130143-01-0

25074887

Ac-D-2-Nal-D-Phe(4-Cl)-β-(3-pyridyl)-D-Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2|Ac-D-2-Nal-D-Phe(4-Cl)-β-(3-pyridyl)-D-Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2

Typically exists as solid at room temperature

1.4±0.1 g/cm3

1.668

2.69

16

16

38

102

2840

10

CC(C[C@H](NC([C@H](NC([C@@H](NC([C@@H](NC([C@H](NC([C@H](NC([C@H](NC(C)=O)CC1=CC2=CC=CC=C2C=C1)=O)CC3=CC=C(Cl)C=C3)=O)CC4=CN=CC=C4)=O)CO)=O)CC5=CC=C(O)C=C5)=O)CCCNC(N)=O)=O)C(N[C@H](C(N6CCC[C@H]6C(N[C@@H](C(N)=O)C)=O)=O)CCCNC(N)=N)=O)C

SBNPWPIBESPSIF-MHWMIDJBSA-N

InChI=1S/C70H92ClN17O14/c1-39(2)31-52(61(94)82-51(15-9-28-77-69(73)74)68(101)88-30-10-16-58(88)67(100)79-40(3)59(72)92)83-60(93)50(14-8-29-78-70(75)102)81-63(96)54(34-43-20-25-49(91)26-21-43)86-66(99)57(38-89)87-65(98)56(36-45-11-7-27-76-37-45)85-64(97)55(33-42-18-23-48(71)24-19-42)84-62(95)53(80-41(4)90)35-44-17-22-46-12-5-6-13-47(46)32-44/h5-7,11-13,17-27,32,37,39-40,50-58,89,91H,8-10,14-16,28-31,33-36,38H2,1-4H3,(H2,72,92)(H,79,100)(H,80,90)(H,81,96)(H,82,94)(H,83,93)(H,84,95)(H,85,97)(H,86,99)(H,87,98)(H4,73,74,77)(H3,75,78,102)/t40-,50-,51+,52+,53-,54+,55-,56-,57+,58+/m1/s1

(S)-1-(((R)-2-((S)-2-((S)-2-((R)-2-((R)-2-((R)-2-acetamido-3-(naphthalen-2-yl)propanamido)-3-(4-chlorophenyl)propanamido)-3-(pyridin-3-yl)propanamido)-3-hydroxypropanamido)-3-(4-hydroxyphenyl)propanamido)-5-ureidopentanoyl)-L-leucyl-L-arginyl)-N-((R)-1-amino-1-oxopropan-2-yl)pyrrolidine-2-carboxamide

CD 20761 D-20761 D20761NS-75A NS 75A SB-075 acetate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)