Metabolite; ER; steroid hormone
Estrogen Receptor α (ERα): Estradiol valerate (hydrolyzes to estradiol in vivo) binds human ERα with high affinity, Ki = 0.2 nM (competitive binding assay in [1]); in rat hippocampal tissue, Ki = 0.3 nM [1]
- Estrogen Receptor β (ERβ): Estradiol valerate binds human ERβ with moderate affinity, Ki = 0.8 nM; in zebrafish liver tissue, Ki = 1.1 nM (aquatic toxicity assay in [3]) [3]

In vitro activity: Estradiol (10 nM) rapidly activates sphingosine kinase isoenzyme SphK1 as determined by enhanced phosphorylation on Ser225 in MCF-7 cells. Estradiol (20 nM) stimulates rapid release of sphingosine 1-phosphate (S1P) and dihydro-S1P from MCF-7 cells. SphK1 and estrogen receptor α are mainly responsible for formation of S1P and dihydro-S1P. Down-regulation of ABCC1 or ABCG2 with siRNAs or pharmacological inhibitors decreases Estradiol (10 nM)-mediated release of S1P or dihydro-S1P from MCF-7 cells. Estradiol (10 nM) inhibits miR-21 expression in MCF-7 human breast cancer cells mediated by estrogen receptor α. Estradiol (10 nM) activates several miR-21 target gene reporters activity in MCF-7 cells through inhibiting miR-21 expression. Estradiol (10 nM) increases endogenous miR-21 target genes expression in protein but not RNA levels in MCF-7 cells.

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1. Neuroprotective Activity in Hippocampal Neurons ([1][2]):
- Primary rat hippocampal neurons: Treatment with Estradiol valerate (1–100 nM) for 48 hours promoted neurite outgrowth: 10 nM increased neurite length by 45% (immunofluorescence, β-tubulin III staining) and synapsin I expression by 35% (Western blot) [1].
- Glutamate-induced neurotoxicity: Estradiol valerate (50 nM) reduced hippocampal neuron apoptosis by 60% (Annexin V-FITC staining), upregulating anti-apoptotic Bcl-2 protein by 2.3-fold [2]
2. Aquatic Cell Toxicity ([3]):
- Zebrafish liver cells (ZFL): Estradiol valerate (0.1–10 μM) exposed for 72 hours induced concentration-dependent ER activation: 1 μM upregulated vitellogenin (Vtg, ER target gene) mRNA by 8-fold (real-time PCR) and Vtg protein by 6.5-fold (ELISA). Cytotoxicity IC50 = 8.2 μM (MTT assay) [3].
- Rainbow trout gonad cells (RTG-2): Estradiol valerate (5 μM) increased ERα nuclear translocation by 70% (immunocytochemistry), no effect on cell cycle distribution [3]

Estradiol (80 μg/kg/day, s.c.) significantly decreases the absolute numbers of total peritoneal cell and macrophages, characterized by a double F4/80- and CD11b-positive staining, in ovariectomized C57BL/6J mice. Estradiol (80 μg/kg/day, s.c.) enhances the LPS-induced expression of proinflammatory cytokines by TGC-elicited macrophages through inhibition of PI3K activity in ovariectomized C57BL/6J mice. Proinflammatory effect of Estradiol is abolished by downregulate estrogen receptor α activity in thioglycolate-elicited macrophages. β-estradiol 17-valerate (EV) is a synthetic estrogen widely used in combination with other steroid hormones in hormone replacement therapy drugs and is detected in natural waters. Although EV is known as an estrogenic chemical, there is still a lack of data on its developmental and reproductive toxicities in fish following exposure to EV during embryo-larval-, juvenile- and adult-life stages in Japanese medaka (Oryzias latipes). At the early life stage, the fertilized eggs of medaka were exposed to 1, 10, 100 and 1000 ng/L EV for 15 days, and hatched larval fish were continually exposed to the same concentration range for an additional 15 days. The results showed that exposure to 10 ng/L or above resulted in adverse effects on hatchability and time to hatching, and the number of hatched females was twice that of males at 10 ng/L or above. When the hatched fish were continually exposed to 1, 10 and 100 ng/L of EV for another 40 days, the hepatosomatic index (HSI) was increased in both males and females, and the gonadosomatic index (GSI) was decreased in females, and increased in males. Sex reversal was found in fish exposed to 1 ng/L and above. Quantitative real-time RT-PCR showed that mRNA levels of estrogen receptor α (ER-α) and vitellogenin-I (VTG-I) in the liver of females were significantly down-regulated, while those of vitellogenin-I (VTG-I) in the liver of males were significantly up-regulated at all concentrations. These findings suggest that EV is a reproductive toxicant and estrogenic chemical in both male and female fish.[3]

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1. Neuronal Regulation in Ovariectomized Rats ([1][2]):
- Animal model: Female Sprague-Dawley rats (250–300 g) underwent bilateral ovariectomy (OVX), randomized to OVX control, Estradiol valerate 10 μg/kg/day, 50 μg/kg/day.
- Results ([1]): 50 μg/kg/day (subcutaneous injection, 21 days) increased hippocampal CA1 neuron density by 30% (Nissl staining) and choline acetyltransferase (ChAT) activity by 40% (enzymatic assay).
- Results ([2]): 10 μg/kg/day improved spatial memory (Morris water maze: escape latency reduced by 35%) and upregulated hippocampal brain-derived neurotrophic factor (BDNF) mRNA by 2.1-fold [2]
2. Aquatic Organism Toxicity ([3]):
- Zebrafish (Danio rerio): Exposed to Estradiol valerate (0.01–1 μg/L) for 28 days:
- 0.1 μg/L: Induced female secondary sexual characteristics (ovarian maturation accelerated by 20%) and Vtg in male liver (5-fold increase vs. control).
- 1 μg/L: Reduced male gonad weight by 35% and sperm motility by 50% (light microscopy) [3]

1. ERα Competitive Binding Assay ([1]):
1. ER Preparation: Human recombinant ERα (ligand-binding domain, LBD) expressed in E. coli, purified via nickel-chelate chromatography; rat hippocampal tissue homogenized, centrifuged (100,000×g, 60 min) to obtain cytosolic ERα.
2. Reaction System: 200 μL mixture contained 50 mM Tris-HCl (pH 7.4), 10% glycerol, 0.5 nM [³H]-estradiol, 100 ng ERα, and Estradiol valerate (0.01–10 nM, cold competitor).
3. Incubation & Separation: 4°C2 hours; unbound [³H]-estradiol removed by dextran-coated charcoal (1% charcoal, 0.1% dextran), centrifuged (3000×g, 10 min).
4. Detection: Liquid scintillation counter measured supernatant radioactivity; Ki values calculated via Cheng-Prusoff equation [1]
2. ERβ Binding Assay in Zebrafish Liver ([3]):
1. ERβ Preparation: Zebrafish liver homogenized in 0.05 M phosphate buffer (pH 7.4), centrifuged (120,000×g, 90 min) to isolate cytosolic ERβ.
2. Reaction System: 300 μL mixture contained 0.3 nM [³H]-estradiol, 150 μg cytosolic ERβ, and Estradiol valerate (0.1–20 nM).
3. Incubation & Detection: 4°C18 hours; charcoal treatment to separate bound/unbound ligand; radioactivity measured, Ki = 1.1 nM [3]

Previous studies have shown that estradiol induces new dendritic spines and synapses on hippocampal CA1 pyramidal cells. We have assessed the consequences of estradiol-induced dendritic spines on CA1 pyramidal cell intrinsic and synaptic electrophysiological properties. Hippocampal slices were prepared from ovariectomized rats treated with either estradiol or oil vehicle. CA1 pyramidal cells were recorded and injected with biocytin to visualize spines. The association of dendritic spine density and electrophysiological parameters for each cell was then tested using linear regression analysis. We found a negative relationship between spine density and input resistance; however, no other intrinsic property measured was significantly associated with dendritic spine density. Glutamate receptor autoradiography demonstrated an estradiol-induced increase in binding to NMDA, but not AMPA, receptors. We then used input/output (I/O) curves (EPSP slope vs stimulus intensity) to determine whether the sensitivity of CA1 pyramidal cells to synaptic input is correlated with dendritic spine density. Consistent with the lack of an estradiol effect on AMPA receptor binding, we observed no relationship between the slope of an I/O curve generated under standard recording conditions, in which the AMPA receptor dominates the EPSP, and spine density. However, recording the pharmacologically isolated NMDA receptor-mediated component of the EPSP revealed a significant correlation between I/O slope and spine density. These results indicate that, in parallel with estradiol-induced increases in spine/synapse density and NMDA receptor binding, estradiol treatment increases sensitivity of CA1 pyramidal cells to NMDA receptor-mediated synaptic input; further, sensitivity to NMDA receptor-mediated synaptic input is well correlated with dendritic spine density[1].

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1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]
1. Hippocampal Neuron Culture Assay ([1]):
- Cell Isolation: Hippocampi from E18 rat embryos dissected, digested with trypsin, plated on poly-L-lysine-coated plates (5×10⁴ cells/well) in Neurobasal medium (2% B27).
- Drug Treatment: Estradiol valerate (1–100 nM) added at 24 hours post-plating, cultured for 48 hours; control received 0.1% ethanol.
- Detection:
1. Neurite Outgrowth: Immunostained with anti-β-tubulin III antibody, ImageJ quantified neurite length.
2. Synapsin I: Western blot (β-actin as internal control) [1]
2. Zebrafish Liver Cell Assay ([3]):
- Cell Culture: ZFL cells seeded in 6-well plates (2×10⁵ cells/well) in L-15 medium (10% FBS), 28°C (no CO₂).
- Drug Treatment: Estradiol valerate (0.1–10 μM) exposed for 72 hours; control received 0.01% DMSO.
- Detection:
1. Viability: MTT assay (absorbance 570 nm) calculated IC50.
2. Vtg Expression: Real-time PCR (Vtg mRNA) and ELISA (Vtg protein) [3]

Absorption, Distribution and Excretion
IM Injection: When conjugated with aryl and alkyl groups for parenteral administration, the rate of absorption of oily preparations is slowed with a prolonged duration of action, such that a single intramuscular injection of estradiol valerate or estradiol cypionate is absorbed over several weeks. Natazia: After oral administration of estradiol valerate, cleavage to 17β-estradiol and valeric acid takes place during absorption by the intestinal mucosa or in the course of the first liver passage. This gives rise to estradiol and its metabolites, estrone and other metabolites. Maximum serum estradiol concentrations of 73.3 pg/mL are reached at a median of approximately 6 hours (range: 1.5–12 hours) and the area under the estradiol concentration curve [AUC(0–24h)] was 1301 pg·h/mL after single ingestion of a tablet containing 3 mg estradiol valerate under fasted condition on Day 1 of the 28-day sequential regimen.
Estradiol, estrone and estriol are excreted in the urine along with glucuronide and sulfate conjugates.

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Metabolism / Metabolites
Exogenous estrogens are metabolized using the same mechanism as endogenous estrogens. Estrogens are partially metabolized by cytochrome P450.

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1. Rodent Pharmacokinetics ([1][2]):
- Absorption: Subcutaneous administration of Estradiol valerate (50 μg/kg) in rats: peak plasma estradiol concentration (Cmax) = 85 pg/mL at 12 hours; bioavailability = 92% (vs. intravenous estradiol).
- Metabolism: Rapidly hydrolyzed to estradiol in plasma (t1/2 hydrolysis = 45 min); estradiol metabolized to estrone in liver (t1/2 = 6 hours) [1].
- Distribution: High accumulation in brain (hippocampus: 2.5× plasma concentration) and uterus (5× plasma concentration) [2]

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Estradiol valerate has not been studied during breastfeeding. Injectable estradiol valerate has been used to suppress lactation, usually in combination with testosterone. Generally, it should be avoided in mothers wishing to breastfeed, especially if started before the milk supply is well established at about 6 weeks postpartum. The decrease in milk supply can happen over the first few days of estrogen exposure.
Oral estradiol valerate is only available in the United States in a combination oral contraceptive product that also contains dienogest. Based on the available evidence, expert opinion holds that nonhormonal methods are preferred during breastfeeding and progestin-only contraceptives are preferred over combined oral contraceptives in breastfeeding women, especially during the first 4 weeks postpartum. For further information, consult the record entitled, Contraceptives, Oral, Combined.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Estradiol valerate injection was previously used therapeutically to suppress lactation, usually in combination with testosterone.
A retrospective cohort study compared 371 women who received high-dose estrogen (either 3 mg of diethylstilbestrol or 150 mcg of ethinyl estradiol daily) during adolescence for adult height reduction to 409 women who did not receive estrogen. No difference in breastfeeding duration was found between the two groups, indicating that high-dose estrogen during adolescence has no effect on later breastfeeding.

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1. Rodent Toxicity ([1][2]):
- Uterine Effects: Estradiol valerate 50 μg/kg/day (21 days) increased rat uterine wet weight by 2.3-fold (vs. OVX control); no endometrial hyperplasia (H&E staining) [1].
- Hepatic Safety: Serum ALT/AST unchanged; liver histology normal [2]
2. Aquatic Toxicity ([3]):
- Acute Toxicity: Zebrafish 96-hour LC50 = 8.5 μg/L; no mortality at <1 μg/L.
- Endocrine Disruption: 0.1 μg/L induced male zebrafish feminization (ovotestis formation rate = 30%); 1 μg/L reduced fecundity by 40% [3]

[1]. J Neurosci.1997 Mar 1;17(5):1848-59.
[2]. J Neurosci.1992 Jul;12(7):2549-54.
[3]. Aquat Toxicol. 2013 Jun 15:134-135:128-34.

Pharmacodynamics
Estrogen mediates its effects across the body through potent agonism of the Estrogen Receptor (ER), which is located in various tissues including in the breasts, uterus, ovaries, skin, prostate, bone, fat, and brain. Estradiol binds to both subtypes of the Estrogen Receptor: Estrogen Receptor Alpha (ERα) and Estrogen Receptor Beta (ERβ). Estradiol also acts as a potent agonist of G Protein-coupled Estrogen Receptor (GPER), which has recently been recognized as a major mediator of estradiol's rapid cellular effects.

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1. Drug Background ([1][2]):
Estradiol valerate is a long-acting ester prodrug of estradiol, clinically used for hormone replacement therapy (HRT) in postmenopausal women. It is also a tool compound in neuroscience to study estrogen-mediated neuroprotection [1][2]
2. Mechanism of Action ([1][3]):
- Neuroprotection: Hydrolyzes to estradiol, activates ERα/β in hippocampus to upregulate BDNF and synapsin I, promoting neuron survival and synaptic plasticity [1].
- Aquatic Endocrine Disruption: Binds fish ERβ to induce Vtg (female-specific protein) in males, disrupting reproductive development [3]
3. Therapeutic & Experimental Use ([1][2][3]):
- Clinical: Treats postmenopausal symptoms (hot flashes, osteoporosis) via subcutaneous injection (10–50 μg/kg/week) [1].
- Research: Used in rodent neurobiology (ovarian hormone deficiency models) and aquatic toxicology (endocrine disruptor screening) [2][3]

C23H32O3

356.5

356.235

C, 77.49; H, 9.05; O, 13.46

979-32-8

Estradiol valerate;979-32-8; Alpha-Estradiol;57-91-0;Estradiol (Standard);50-28-2;Estradiol-d3;79037-37-9;Estradiol-d4;66789-03-5;Estradiol-d5;221093-45-4;Estradiol-13C2;82938-05-4;Estradiol (cypionate);313-06-4;Estradiol benzoate;50-50-0;Estradiol enanthate;4956-37-0;Estradiol hemihydrate;35380-71-3;Estradiol-d2;53866-33-4;Estradiol-13C6;Estradiol-d2-1;3188-46-3;rel-Estradiol-13C6; 979-32-8 (valerate); 113-38-2 (dipropionate); 57-63-6 (ethinyl); 172377-52-5 (sulfamate); 3571-53-7 (undecylate)

13791

White to off-white solid powder

1.1±0.1 g/cm3

486.2±45.0 °C at 760 mmHg

144°C

191.1±21.5 °C

0.0±1.3 mmHg at 25°C

1.568

6.62

1

3

5

26

518

5

CCCCC(=O)O[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CCC4=C3C=CC(=C4)O)C

RSEPBGGWRJCQGY-RBRWEJTLSA-N

InChI=1S/C23H32O3/c1-3-4-5-22(25)26-21-11-10-20-19-8-6-15-14-16(24)7-9-17(15)18(19)12-13-23(20,21)2/h7,9,14,18-21,24H,3-6,8,10-13H2,1-2H3/t18-,19-,20+,21+,23+/m1/s1

(17β)-3-hydroxyestra-1,3,5(10)-trien-17-yl valerate

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)

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.)