ER/estrogen-receptor; Clomiphene binds to muscarinic cholinergic receptors and calcium channel blocker binding sites in cell membranes [1]

在来自多种组织的膜制剂中，氯米芬（Clomiphene）抑制放射性标记的毒蕈碱配体（如奎宁环基苄酸盐）和钙通道阻滞剂（如尼群地平）与其各自结合位点的结合。这种抑制具有浓度依赖性，表明其与这些位点存在竞争性或非竞争性相互作用 [1]

对雄性小鼠进行围产期氯米芬（Clomiphene）处理（从妊娠第12天至出生后第10天，给孕鼠每日注射100 μg），可改变其性取向，表现为交配行为和偏好测试中的变化。与对照组相比，处理后的雄性小鼠对雌性小鼠的偏好降低 [2]
在临床环境中，氯米芬（Clomiphene）用于无排卵性不孕女性的促排卵治疗。它刺激垂体释放促卵泡生成素（FSH）和促黄体生成素（LH），从而促进卵泡发育和排卵。据报道，其排卵率为70-80%，妊娠率为30-40% [3]

制备组织（如脑、心脏）的膜组分，在不同浓度的氯米芬（Clomiphene）存在下，与放射性标记的毒蕈碱配体或钙通道阻滞剂共同孵育。孵育后，通过过滤或离心分离结合态和游离态放射性配体。测量结合态放射性配体的量，并生成抑制曲线以评估氯米芬（Clomiphene）与结合位点的相互作用 [1]

For perinatal studies, pregnant mice receive Clomiphene via subcutaneous injection at a dose of 100 μg/day from gestational day 12 to postnatal day 10. Control groups receive vehicle injections. Offspring male mice are tested for sexual orientation using behavioral assays (e.g., preference for female mice vs. male mice) after reaching adulthood [2]

Absorption, Distribution and Excretion
Based on early studies of clomiphene citrate labeled with 14C, this drug is readily absorbed orally in humans. Based on early studies of clomiphene citrate labeled with 14C, this drug is readily absorbed orally in humans and is primarily excreted via feces. The average urinary excretion rate is approximately 8%, and the fecal excretion rate is approximately 42%. Subcutaneous injection of clomiphene citrate labeled with 14C…distributed in the tissues of female newborn guinea pigs…estrogen-responsive tissues showed a high affinity for 14C. (14)C levels remained stable in the uterus…decreased in the ovaries and plasma…increased in the adrenal glands. /clomiphene citrate/ Approximately half of the ingested dose was excreted within five days; trace amounts of the drug remained in the feces for up to six weeks after administration. /Clomiphene Citrate/
Clomiphene is well absorbed after oral administration. The drug and its metabolites are primarily excreted in feces, with a small amount excreted in urine. The relatively long plasma half-life (approximately 5 to 7 days) is mainly due to plasma protein binding, enterohepatic circulation, and accumulation in adipose tissue. Long-lived active metabolites may also be produced.

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Metabolism/Metabolites
Liver
Incubation of the nonsteroidal anti-estrogenic clomiphene with rat liver microsomes resulted in the formation of 4-hydroxy, N-deethyl, and N-oxide metabolites, in stark contrast to previous similar experiments using rabbit microsomes, where only the first two metabolites were detected. No urinary excretion of the drug or its metabolites was detected after oral administration of clomiphene. 4-hydroxyclomiphene was the only detectable elimination product in the fecal extract.
Liver

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Biological Half-Life
5-7 days

Toxicity Summary
Clomiphene possesses both estrogenic and anti-estrogenic properties, but its exact mechanism of action remains unclear. Clomiphene appears to stimulate the release of gonadotropins (follicle-stimulating hormone (FSH) and luteinizing hormone (LH)), thereby promoting follicular maturation, ovulation, and corpus luteum development and function, ultimately leading to pregnancy. The release of gonadotropins may be due to direct stimulation of the hypothalamic-pituitary axis, or it may be due to clomiphene competing with endogenous estrogens from the uterus, pituitary gland, or hypothalamus, thereby reducing the inhibitory effect of estrogen on the hypothalamic-pituitary axis. Clomiphene has no significant progesterone, androgenic, or anti-androgenic effects and does not appear to interfere with the function of the pituitary-adrenal or pituitary-thyroid axis. Toxicity Data
The acute oral LD50 of clomiphene is 1700 mg/kg in mice and 5750 mg/kg in rats. The toxic dose in humans is unknown. There are no reports of acute overdose of clomiphene. During clomiphene treatment, if the dose exceeds the recommended dose, overdose symptoms may occur, including nausea, vomiting, vasomotor flushing, blurred vision, spots or flashes of light in front of the eyes, scotomas, enlarged ovaries, and pelvic or abdominal pain. Hepatotoxicity: Information on serum transaminase levels during clomiphene treatment is limited because clomiphene is usually taken only in low doses for short periods. Although there are a few reports of mild serum enzyme elevations in patients taking clomiphene, there is no conclusive evidence that it causes specific, clinically significant liver damage. Drugs used to treat female infertility often work by stimulating ovarian follicles, which can lead to ovarian hyperstimulation syndrome (OHSS), sometimes accompanied by elevated serum enzymes and even jaundice. This syndrome usually appears 4 to 14 days after ovarian stimulation with gonadotropins or clomiphene and is characterized by abdominal pain, abdominal distension with ascites, enlarged ovaries, and ovarian cysts. Patients may experience significant fluid shifts, leading to hemoconcentration and rapid development of severe ascites and pleural effusion. Liver dysfunction is present in 25% to 40% of patients with ovarian hyperstimulation syndrome (OHSS), typically manifested as mild to moderate elevations in alanine aminotransferase (ALT) and aspartate aminotransferase (AST), but with minimal or no elevation in serum bilirubin and alkaline phosphatase levels. Liver dysfunction usually resolves as OHSS subsides, typically within 2 to 3 weeks of onset. In severe cases, OHSS can be fatal, but death is usually due to dehydration, shock, and sepsis, rather than liver failure. In typical cases of abnormal liver enzymes, liver histology shows nonspecific changes, including sinusoidal dilatation, mild fatty deposits, and focal inflammatory infiltration dominated by macrophages and lymphocytes. The incidence of OHSS is lower with clomiphene citrate compared to ovulation induction with human chorionic gonadotropin (hCG).
Probability Score: C (Possibly the cause of clinically significant liver damage in ovarian hyperstimulation syndrome).

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Effects during pregnancy and lactation
◉ Summary of medication use during lactation
A small amount of clomiphene was detected in the breast milk of a woman. Multiple studies have found that clomiphene can suppress lactation in women who do not wish to breastfeed. Its mechanism of action appears to be through reducing serum prolactin levels, especially the peak serum prolactin level after stimulation. Clomiphene may interfere with lactation in breastfeeding mothers.
◉ Effects on breastfed infants
A woman taking 2.04 mg/kg clomiphene daily breastfed part-time. She did not observe any adverse reactions in her infant.
◉ Effects on Lactation and Breast Milk
A double-blind study compared the effects of clomiphene daily at 50 mg for 10 days (n = 110), clomiphene daily at 100 mg for 5 days (n = 26), and placebo (n = 41) on inhibiting lactation and relieving pain and breast engorgement in non-lactating postpartum mothers. According to women's reports, both doses of clomiphene were superior to placebo, but the 100 mg daily dose was slightly superior to the 50 mg daily dose.
A study compared the effects of clomiphene daily at 100 mg (n = 60) for 5 days with placebo (n = 30) on inhibiting lactation and relieving breast engorgement symptoms. Physicians observed that starting clomiphene within 12 hours postpartum was superior to starting clomiphene at 12 hours or longer postpartum in all indicators; both treatments were more effective than mechanical measures alone (e.g., breast binding). A randomized trial compared the effects of four treatment regimens on reducing postpartum serum prolactin levels and decreasing lactation: clomiphene 50 mg twice daily for 14 days (n = 15); bromocriptine 2.5 mg twice daily for 14 days (n = 15); diethylstilbestrol 5 mg three times daily for 14 days (n = 15); testosterone propionate 75 mg intramuscularly once (n = 15); and placebo orally three times daily (n = 15). After three days of treatment, serum prolactin levels in the clomiphene group decreased to 65% of baseline, while those in the bromocriptine group decreased to only 35%. Clomiphene was also less effective than bromocriptine in suppressing lactation and relieving breast engorgement. Another study compared the effects of starting clomiphene 100 mg/day on the first postpartum day for 7 days (n = 10) with placebo (n = 12). The results showed that clomiphene was no more effective than placebo in suppressing lactation or lowering serum prolactin levels. Women who did not wish to breastfeed during the first week postpartum were either treated with clomiphene 50 mg/day twice daily (n=10) or a placebo (n=10). Women taking clomiphene did not experience elevated serum prolactin levels above baseline during breast pump use; women taking the placebo experienced normal post-stimulation increases in serum prolactin. A total of 80 postpartum women participated in the study. Forty women took clomiphene 50 mg/day twice daily for five consecutive days starting from day one postpartum; 20 women took clomiphene 50 mg twice daily for five consecutive days starting from day four postpartum; and the remaining 20 women received a placebo. All women taking clomiphene experienced lactation suppression, breast engorgement, discomfort, and decreased serum prolactin levels. For subjects 1 day postpartum, serum prolactin concentrations were significantly lower than baseline levels on day 3; for subjects 4 days postpartum, serum prolactin concentrations were significantly lower than baseline levels on day 5. Placebo did not suppress lactation or lower serum prolactin levels. Adverse Reactions: Some reported adverse reactions to clomiphene include headache, dizziness, exacerbation of psychiatric illness, gynecomastia, testicular tumors, vasomotor flushing, gastrointestinal discomfort, and breast pain. Other common adverse reactions include nausea, vomiting, ovarian enlargement, blurred vision, scotoma, abnormal uterine bleeding, pelvic pain, and hypertriglyceridemia. Some serious adverse reactions to clomiphene include multiple pregnancy, thrombocytopenia, pancreatitis, increased risk of ovarian cancer with long-term use, increased risk of malignant melanoma, severe visual impairment, and liver damage. Ovarian hyperstimulation syndrome (OHSS) has been reported in patients treated with clomiphene citrate for ovulation induction. OHSS can progress rapidly (within 24 hours) and may develop into a medical emergency.

[1]. Interaction of antiestrogens with binding sites for muscarinic cholinergic drugs and calcium channel blockers in cell membranes. Cancer Chemother Pharmacol. 1990;26(4):310-2;
[2]. [Perinatal clomiphene citrate treatment changes sexual orientations of male mice]. Dongwuxue Yanjiu. 2013 Oct;34(5):487-92.
[3]. Ovulation induction with clomifene: a primary care perspective. J Fam Plann Reprod Health Care. 2012;38(1):48-52.

Therapeutic Uses
Female Fertility Agents Veterinary Drugs: Used to induce ovulation in anovulatory female animals and to induce the restoration of normal menstrual cycles in animals with amenorrhea or oligomenorrhea. Multiple Pregnancy Rate: Approximately 8% of pregnancies are induced, six times the normal rate, but lower than with human menopausal gonadotropin (HMG). Multiple pregnancies are almost always twins; large multiple pregnancies are reported in rare cases. About 20% of pregnancies induced by clomiphene citrate result in spontaneous abortion (mostly early abortion), a rate that may be only slightly higher than normal but not higher than in infertile individuals. Clomiphene Citrate Clomiphene citrate has also been used to treat male oligospermia…but the value of this treatment has not been determined. Clomiphene Citrate For more complete data on the therapeutic uses of clomiphene (10 types), please visit the HSDB record page.

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Drug Warnings
Objective signs are rare, but there have been reports of decreased vision, definable scotoma, and altered retinal cell function. Clomiphene Citrate
Other adverse reactions include…headache, breast tenderness, and abdominal distension. Symptoms disappear after discontinuation of the drug. Clomiphene Citrate
Some physicians consider visual abnormalities a contraindication to continued use, while others continue treatment with lower doses. …Clomiphene should not be given to pregnant women; there is no indication for clomiphene treatment once pregnancy occurs. Clomiphene Citrate
Sulfobromophthalein retention may increase. High doses may increase norepinephrine levels.
For more complete data on drug warnings for clomiphene (21 in total), please visit the HSDB records page.

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Pharmacodynamics
Clomiphene (formerly known as clomiphene) is an oral nonsteroidal ovulation-inducing drug belonging to the class of selective estrogen receptor modulators (SERMs). Clomiphene can cause polyovulation, thus increasing the risk of twins. Clomiphene has a lower incidence of ovarian hyperstimulation syndrome compared to purified follicle-stimulating hormone (FSH). There may be risks of ovarian cancer and weight gain. Clomiphene can interact with tissues containing estrogen receptors, including the hypothalamus, pituitary gland, ovaries, endometrium, vagina, and cervix. Clomiphene may compete with estrogen for estrogen receptor binding sites and may delay the replenishment of intracellular estrogen receptors. Clomiphene initiates a series of endocrine events that ultimately lead to a pre-ovulatory gonadotropin surge and subsequent follicle rupture. The first endocrine event in clomiphene treatment is an increase in pituitary gonadotropin release. This initiates steroid production and follicle development, leading to ovarian follicle growth and elevated circulating estradiol levels. After ovulation, plasma progesterone and estradiol levels rise and fall as in a normal ovulatory cycle. Clomiphene is a styrene derivative with anti-estrogenic properties and is commonly used in reproductive medicine. Its ability to interact with muscarinic receptors and calcium channel binding sites suggests that, in addition to its major role in inducing ovulation, there may be potential off-target effects [1][3]

C26H28NOCL

405.95962

405.186

C, 76.92; H, 6.95; Cl, 8.73; N, 3.45; O, 3.94

911-45-5

Clomiphene citrate;50-41-9;Clomifene hydrochloride;57049-00-0;Clomifene-d5 hydrochloride;1346606-66-3

1548953

Typically exists as solid at room temperature

1.104g/cm3

509ºC at 760mmHg

117.25°C

261.6ºC

1.588

6.562

12.47

0

2

9

29

481

0

Cl/C(C1=CC=CC=C1)=C(C2=CC=C(OCCN(CC)CC)C=C2)/C3=CC=CC=C3

GKIRPKYJQBWNGO-OCEACIFDSA-N

InChI=1S/C26H28ClNO/c1-3-28(4-2)19-20-29-24-17-15-22(16-18-24)25(21-11-7-5-8-12-21)26(27)23-13-9-6-10-14-23/h5-18H,3-4,19-20H2,1-2H3/b26-25+

2-[4-[(E)-2-chloro-1,2-diphenylethenyl]phenoxy]-N,N-diethylethanamine

clomiphene; Clomiphene B; 911-45-5; Chlomaphene; Chloramifene; Clomifen; 2-(p-(2-Chloro-1,2-diphenylvinyl)phenoxy)triethylamine; 1HRS458QU2;

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

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。