(5α)-Pregnane-3,20-dione

• CAT Number: R018031
• CAS Number: 566-65-4
• Molecular Formula: C21H32O2
• Molecular Weight: 316.485
• Purity: ≥95%
• Synonyms: 5α-Pregnane-3,20-dione (8CI); 5α-Pregnanedione (7CI); (+)-(5α)-Pregnane-3,20-dione; 3,20-Allopregnanedione; 3,20-Dioxo-5α-pregnane; 5α-Dihydro-P4; 5α-Dihydroprogesterone; NSC 18319
• Storage: RT
• IUPAC Name: (5S,8R,9S,10S,13S,14S,17S)-17-acetyl-10,13-dimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-3-one
• InChI: InChI=1S/C21H32O2/c1-13(22)17-6-7-18-16-5-4-14-12-15(23)8-10-20(14,2)19(16)9-11-21(17,18)3/h14,16-19H,4-12H2,1-3H3/t14-,16-,17+,18-,19-,20-,21+/m0/s1
• InChIKey: XMRPGKVKISIQBV-BJMCWZGWSA-N
• SMILES: CC(=O)C1CCC2C1(CCC3C2CCC4C3(CCC(=O)C4)C)C

5alpha-pregnane-3,20-dione (CAS 566-65-4), a metabolite of progestrone, belongs to the class of organic compounds known as gluco/mineralocorticoids, progestogins and derivatives.It is a C21-steroid hormone that is 5alpha-pregnane substituted by oxo groups at positions 3 and 20. It is a 20-oxo steroid, a C21-steroid hormone and a 3-oxo-5alpha-steroid. It derives from a progesterone. It derives from a hydride of a 5alpha-pregnane.  5alpha-Pregnane-3, 20-dione exists as a solid and is considered to be practically insoluble (in water) and relatively neutral.Within the cell, 5alpha-pregnane-3, 20-dione is primarily located in the membrane (predicted from logP) and cytoplasm. In humans, 5alpha-pregnane-3, 20-dione is involved in the steroidogenesis pathway. 5alpha-Pregnane-3, 20-dione is also involved in several metabolic disorders. Memberane 5alpha-pregnane-3,20-dione receptor was associated with cell proliferation and detachment in MCF-7 breast cancer cell line. 

Reference

1. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4341-5. doi: 10.1073/pnas.71.11.4341.

Characterization of binding components for progesterone and 5alpha-pregnane-3,20-dione in the hamster uterus.Leavitt WW, Grossman CJ.

 

In the hamster uterus, a specific progesterone (pregn-4-ene-3,20-dione) receptor 

has been identified in the cytosol fraction. In the present study, we examined hamster uterine cytosol for the possible existence of specific macromolecules that bind the progesterone metabolite, 5alpha-pregnane-3,20-dione. When cytosol was analyzed by density-gradient centrifugation with sucrose-glycerol gradients and by Scatchard plot analysis of [(3)H]5alpha-pregnane-3,20-dione binding data, there was no evidence of specific binding components for this metabolite. In vivo treatment of proestrous hamsters with unlabeled progesterone, 5alpha-pregnane-3,20-dione, or cortisol for 1 hr revealed that only progesterone caused the depletion of progesterone-receptor sites from the uterine cytosol 

fraction. Incubation of uterine strips which had been preloaded with two different concentrations of [(3)H]-progesterone demonstrated that progesterone was metabolized to 5alpha-pregnane-3,20-dione and to a greater extent to 3alpha-hydroxy-5alpha-pregnan-20-one. The accumulation of 5alpha-pregnane-3,20-dione during progesterone metabolism appeared to be related to the availability of nonspecifically bound hormone. These studies (i) strongly suggest there is no specific receptor system for 5alpha-pregnane-3,20-dione in the uterine cytosol fraction, (ii) confirm the existence of a specific progesterone receptor in uterine cytosol, and (iii) provide evidence that progesterone itself mediates the uterine progestational response via interaction with a specific receptor system.

 

2. Endocrine. 2001 Oct;16(1):7-14. doi: 10.1385/endo:16:1:07.The endogenous progesterone metabolite, 5a-pregnane-3,20-dione, decreases cell-substrate attachment, adhesion plaques, vinculin expression, and polymerized F-actin in MCF-7 breast cancer cells.Wiebe JP(1), Muzia D.

 

Tumorous human breast tissue readily converts progesterone to 5alpha-pregnane-3,20-dione (5alphaP), and this metabolite has been shown to stimulate proliferation and to decrease adhesion of MCF-7 breast cancer cells. To determine the mechanisms of action of 5alphaP on cell adhesion, MCF-7 cells were grown without or with 5alphaP (10(-9)-10(-5) M), and the effects on cell and nuclear morphology, adhesion plaques, vinculin and actin expression, actin polymerization, and microfilament distribution were examined by immunohistochemistry, morphometry (using confocal microscopy and digital computer imaging analysis), and Western blotting. Treatment of cells with 10(-9)-10(-6) M 5alphaP resulted in dose-dependent decreases in cell area, cell-to-cell contacts, and attachment to the substratum, and increases in variation in nuclear area. These changes in the 5alphaP-treated cells were accompanied by decreases in vinculin-containing adhesion plaques, vinculin expression, polymerized actin stress fibers, and decreases in insoluble and 

increases in soluble actin fractions. The results suggest that the observed decreases in adhesion and increases in cell proliferation following 5alphaP treatment may be owing to depolymerization of actin and decreased expression of actin and vinculin. We conclude that the endogenous progesterone metabolite, 5alphaP, may be involved in promoting breast neoplasia and metastasis by affecting adhesion and cytoskeletal molecules.

 

3. Steroids. 1976 Dec;28(6):867-80. doi: 10.1016/0039-128x(76)90036-2.Isolation, identification and quantitation of serum 5alpha-pregnane-3,20-dione and its relationship to progesterone in the pregnant mare.Atkins DT, Harms PG, Sorensen AM Jr, Fleeger JL.

 

5alpha-pregnane-3,20-dione was isolated from pooled pregnant mare serum using Sephadex LH-20 column chromatography and identified by the use of radioimmunoassay, gas-liquid chromatography and gas-liquid chromatography-mass spectrometry analyses. 5beta-pregnane-3,20-dione was not cross-reactive with the radioimmunoassay system and was not detected by gas-liquid chromatography. Peripheral blood levels of progesterone and 5alphs-pregnane-3,20-dione were determined by radioimmunoassay in four Quarter Horse mares for the first 150 days of gestation. Progesterone and 5alpha-pregnane-3,20-dione declined from a range of 6 to 15 ng/ml at day 10 to a range of 2 to 8 ng/ml at day 40. After day 

40 there was an increase in progesterone and 5alpha-pregnane-3,20-dione concentration. Toward the end of 150 days of gestation progesterone tended to decline; whereas, 5alpha-pregnane-3,20-dione levels were maintained or increased to levels as high as 35 ng/ml. Neither source nor function of 5alpha-pregnane-3,20-dione is known.

 

 

4. J Steroid Biochem Mol Biol. 2005 Nov;97(3):278-88. doi: 10.1016/j.jsbmb.2005.05.014. Epub 2005 Sep 9.Membrane 5alpha-pregnane-3,20-dione (5alphaP) receptors in MCF-7 and MCF-10A breast cancer cells are up-regulated by estradiol and 5alphaP and down-regulated by the progesterone metabolites, 3alpha-dihydroprogesterone and 20alpha -dihydroprogesterone, with associated changes in cell proliferation and detachment.Pawlak KJ(1), Zhang G, Wiebe JP.

 

Previous studies have shown that the progesterone metabolite, 5alpha-pregnane-3,20-dione (5alphaP), exhibits mitogenic and metastatic activity in breast cell lines and that specific, high affinity receptors for 5alphaP are located in the plasma membrane fractions of tumorigenic (ER/PR-positive) MCF-7 cells. The aim of this study was to determine the effects of the mitogenic (estradiol; 5alphaP) and anti-mitogenic (3alpha-hydroxy-4-pregnen-20-one, 3alphaHP; 20alpha-hydroxy-4-pregnen-3-one, 20alphaHP) endogenous steroid hormones on 5alphaP receptor (5alphaP-R) numbers and on cell proliferation and adhesion of MCF-7 and MCF-10A cells. Exposure of MCF-7 cells for 24h to estradiol or 5alphaP resulted in significant (p < 0.05-0.001) dose-dependent increases in 5alphaP-R levels. Conversely, treatment with 3alphaHP or 20alphaHP resulted in significant (p <0.05-0.01) dose-dependent decreases in 5alphaP-R levels. Treatment with one mitogenic and one anti-mitogenic hormone resulted in inhibition of the mitogen-induced increases, whereas treatment with two mitogenic or two anti-mitogenic hormones resulted in additive effects on 5alphaP-R numbers. Treatments with cycloheximide and actinomycin D indicate that changes in 5alphaP-R levels depend upon transcription and translation. The non-tumorigenic breast cell line, MCF-10A, was also shown to posses specific, high affinity plasma membrane receptors for 5alphaP that were up-regulated by estradiol and 5alphaP and down-regulated by 3alphaHP. Estradiol binding was demonstrated in MCF-10A cell membrane fractions and may explain the estradiol action in these cells that lack intracellular ER. In both MCF-7 and MCF-10A cells, the increases in 5alphaP-R due to estradiol or 5alphaP, and decreases due to 3alphaHP or 20alphaHP correlate with respective increases and decreases in cell proliferation as well as detachment. These results show distribution of 5alphaP-R in several cell types and they provide further evidence of the significance of progesterone metabolites and their novel membrane-associated receptors in breast cancer stimulation and control. The findings that 3alphaHP and 20alphaHP down-regulate 5alphaP-R and suppress mitogenic and metastatic activity suggest that these endogenous anti-mitogenic progesterone metabolites deserve considerations in designing new breast cancer therapeutic agents.