Kovács, Katalin

Matkovics, Béla

Rakonczay, Zoltán

English

Kovács Katalin

Rakonczay Zoltán

Matkovics Béla

University of Szeged

STEROIDS, xxra. Synthesis of 2- and 4-Hydroxy and 2,4-Dihydroxy Derivatives of Estrone and Estradiol By KATALIN KOVÁCS, Z. RAKONCZAY AND B. MATKOVICS Biochemical and Genetical Groups, Attila József University, Szeged (Received May 19, ¡973) The following hydroxy derivatives of estrone and estradiol were prepared during this work1: 2,3-dihydroxyestrone, 2,3-dihydroxyestradiol, 3,4-dihydroxyestrone, 3,4-dihydroxyestradiol, 2,3,4-tri-hydroxyestrone and 2,3,4-trihydroxyestradiol. The syntheses were performed via the correspond-ing mono and dinitro derivatives, and all intermediates were isolated and described. A number of authors have dealt with the direct synthesis of the group of compounds mentioned in the title [1—4], while others have strived to synthesize the monomethoxy derivatives [5] in. order to identify the methoxy intermediates occurring among natural estrogens [6—7]. As regards the hydroxylated estrogens accumulating during pregnancy, many authors attribute importance exclusively to the compounds mentioned, but primarily to 2-hydroxyestrone [8]. In the case of estrogens bound to protein, and possibly to glutathion, on the other hand, the important intermediate role of these compounds [9—11] is assumed. Valuable results have been obtained recently on the isolation of this group of compounds, and on the study of their properties and role in the intermediate metabolism [12, 13]. The synthesis of these compounds has already been dealt with for purposes of identification of the derivatives obtained in previous non-specific enzymatic hydroxylation experiments [14]. 1 Abbreviations used: 2-nitroestrone means 2-nitro-3-hydroxy-l,3,5(10)-triene-17-one; 2-aminoestrone and 2-hydroxyestrone mean the 2-amino and 2-hydroxy derivatives of this same compound; similarly, 2-nitroestradiol means 2-nitro-3,17B-dihydroxyestra-l,3,5(10)-triene, while 2-aminoestradiol and 2-hydroxyestradiol mean the corresponding 2-amino and 2-hydroxy derivatives, and so on. 288 k a t a l i n k o v a c s , z. r a k o n c z a y a n d b. m a t k o v i c s Experimental2 2- and 4-nitroestrones and 2,4-dinitroestrone were prepared and purified as described by WERBIN and HOLOWAY [1]. The reduction of mono- and dinitroestrones was carried out with sodium borohydride to the corresponding estradiols, just as for the mono- and diamino aind the mono- and dihydroxy substituted derivatives, by the method of PATTON [4]. In the identification an effort was made to attain the chemical and physico-chemical properties for the compounds described in the literature, and also to obtain materials which were chromatographically (TLC) completely pure by running them in a number of well-known solvent pairs. Separations were performed under the usual conditions, on silica gel adsorbent prepared according to Stahl (Merck or Reanal) in an ascending system.3 2-Nitroestrone (I) [1]. M.p.: 178—80°C (Lit.: 183.5—184 or 155°C). Anal.: Calc.: C1 8H2 104N C 68.55 H 6.71 N4.44; Found: C 68.65 H 6.70 H4.41. [a]D = = + 3 0 + 1 ° (c= 1.0; CHC13). 3350(OH), 1750, 1640(CO), 1530 (Ar-NO,), 880, 760, 650 (C-N02) cm-1 . (The IR spectra were taken with a UNICAM SP 1000 spectrophotometer in all cases.) . 2-Nitroestradiol (II) [4]. The sodium borohydride reduction of I save the diol. M.p.: 162—3°C (Lit.: 167—8°C). Anal.: Calc.: C1 8H2 304N C 6 8 . l l H 7.30 N 4.41; Found: C 68.15 H 7.23 N 4.48. [a]D = +160 + 2° (c= 1.0; CHC13). v ^ 3490, 3300, 2950 (OH), 1630, 1525 (Ar-NOa), 965, 990, 760 (C-N02) cm"1 . 2-Aminoestrone (III). It was reduced to amine by the method of KRAYCHY and GALLAGHER [15] with an aqueous solution of alkaline sodium hydrosulphite (Na2S204). The yield was very good. M.p.: 192—93°C (Lit.: 220°C). Anal.: Calc.: C1 8H2 302N C 75.75 H 8.14 N4.91; Found: C 75.80 H8.18 N4.67. [oe]D = - 4 0 ± 2 ° (c=0.5 ; CHCI3). v £ i 3500, 3390, 3350 (NH2, OH), 1730 (CO), 1290 (NH2) cm- 1 . 2-Aminoestradiol (IV). Sodium borohydride reduction of an alkaline methanolic solution of III led to IV. M.p.: 188—90°C. Anal.: Calc.: C18H>50„N C 75.23 H 8.76 N 4.87; Found: C 75.30 H 8.80 N4.72. [a]D = - 1 3 0 + 2° (c = 0.1; CHC13). v™r 3390, 3320, 2950 (NH2 , OH), .1290 1220 (NH2) cm"1 . 2-Hydroxyestrone (V). Prepared from III by diazotation and boiling the diazo compound with urea (by a combination of the methods of NIEDERL and VOGEL [2] and KRAYCHY and GALLAGHER [15]). The 2-hydroxyestrone formed was purified from the by-products by preparative thick layer chromatography on a silica gel adsorbent mentioned before. The end-product of the above reaction was dissolved in C H C 1 3 and transferred in a band to a 500 N thick adsorbent layer. The plate was run in a 1:2:2 mixture of acetone—cyclohexane—chloroform. The identified band, with the appropriate R,̂  value, was scraped off together with the adsorbent. The adsorbent containing the substance was repeatedly extracted with acetone—chloroform (1:2). The adsorbent was removed by filtration on a G5 glass filter and the solvent 2 Melting points are uncorrected. 3 We wish to publish the detailed TLC properties of the compounds in the near future else-where. STEROIDS, XXIII. DERIVATIVES OFESTRONE AND ESTRADIOL 289 was evaporated in N2 atmosphere. (As formation of quinone was observed, it is very important to avoid contact of the material with air as far as possible.) The product thus isolated was pure by TLC. M.p.: 116—18 °C (Lit.: 200°C). Anal.: Calc.: C18H2203 С 75.52 H7.74; Found: С 75.60 H 7.80. [<x]D= +190+2° (c= 1.0; EtOH). v™'x 3350 (OH), 1730, 1710 (CO), 1610, 1290 (Ar) cm"1 . 2-Hydroxyestradiol (VI). Prepared by the previously mentioned reduction of V. M.p.: 194—96 °C. Anal.: Calc.: C18H2403 С 75.00 H8.39; Found: С 75.05 H 8.43. [ a t = +110±2° (c=0 .1 ; EtOH). v ^ 3 4 0 0 ( O H ) , 1510,1450, 1260(Ar)cm-1. 4-Nitroestrone ( VII). M.p.: 277—78 °C (Lit.: 279—80 °C or 258 °C). Anal.: Calc.: C1 8H2 104N (as for I); Found: С68.60 H 6.78 N4.41. [a]D = - 2 9 9 + 2° (c=0.3; CHC13). v™ 3350 (OH), 1730 (CO), 1530 (Ar-N02), 1030, 825, 800 (C-N02) cm - 1 . 4-Nitroestradiol (VIII). Prepared from VII by the method used for II. M.p.: 192—93 °C (Lit.: 220°C). Anal.: Calc.: (as for II); Found: С 68.08 H 7.20 N4.72. MD — + 2 5 0 ± 2 ° (c=0.1; CHC13). 3200, 2950 (OH), 1530 (Ar-NO,), 1050, 825 (C-N02) cm-1 . 4-Aminoestrone (IX). Prepared from VII, analogously as for III. M.p.: 250—52 °C. Anal. :_Calc.: (as for III); Found: С 75.70 H 8.09 N 4.67. [a]0 = + 5 0 + 1 ° (c=0.1; dimethylsulphoxide) 4-Aminoestradiol (X). Prepared from IX as for III and IV. M.p.: 266—68 °C.. Anal.: Calc.: (as for IV); Found: С 75.20 H 8.72 N4.72. [a]D = - 9 0 ± 2 ° (c=0.1; dimethylsulphoxide). 3390, 3320, 2950 (OH, NH2), 1620, 1300, 800 (NH2) cm"1 . 4-Hydroxyestrone (XI). Prepared from IX by the combined method given for V. M.p.: 233—35 °C. Anal.: Calc.': (as for V); Found: С 75.58 H 7.78. MD = = + 100±2° (c=0.1; EtOH). v™rx 3400, 3350 (OH), 1740 (CO), 1290 (Ar) cm-1 . - 4-Hydroxyestradiol (XII). Prepared from XI as for VI. M.p. : 168—70 °C. Anal.: Calc.: (as for VI); Found: С 75.04 H 8.40. MD = +140+2° (c=0.1; EtOH). vS i 3400 (OH), 1510, 1250 (Ar) cm"1 . 2,4-Dinitroestrone (XIII). Prepared as described by WERBIN and HOLOWAY [1]. M.p.: 185—87.5 °C. Anal.: Calc.: C18H20O6N2 С 59.99 H 5.59 N7.78; Found: С 60.05 H 5.62 N8.10. [a]D = + 120±2° (c=1.0; CHC13). vSSi 3350, 3320 (OH), 1750 (CO), 1730, 1535 (Ar-N02), 1320 (N02), '750 (C-NOa) cm-1 . 2,4-Dinitroestradiol (XIV). Prepared from XIII similarly as previously. M.p.: 263—65 °C. Anal.: Calc.: C18H2206N2 С 59.11 H6.12 N7.78; Found: С 59.20 H 6.18 N8.10. [<z]0 = + 100±2° (c=0.1; CHC13). v£Ji 3220,2940 (OH), 1320(N02), 800, 745 (C-NOa) cm"1. 2,4-Diaminoestrone (XV). Prepared as above, by reduction, from XIII. M.p.: 207—9 °C. Anal.: Calc.: C18H2402N2 С 71.97 H 8.05 N9.03; Found: С 80.02 H 8.10 N9.40. Мд = + 2 0 0 ±2° (c=0.1; MeOH). 3390, 3350 (NH2 , OH), 1745 (CO), 1615, 1290 (NH2) cm"1 . 2,4-Diaminoestradiol (XVI). Prepared from XV by further reduction. M.p.: 198—200°C. Anal.: Calc.: C18H2602N C71.52 H 8.66 N9.26; Found: C71.57 2 9 0 k a t a l i n k o v á c s , z . r a k o n c z a y a n d b . m a t k o v i c s H 8.60 N9.40. [ a ] D = + 1 6 0 + 2 ° (c = 0.1; dimethylsulphoxide). 3380, 3320 (NH2 , OH), 1290 (NH2) cm - 1 . 2,4-Dihydroxyestrone (XVJI). Prepared from XV in accordance with the above. M.p.: 228—30°C. Anal.: Calc.: C18H2204 C 71.51 H 7.33; Found: C 71.48 H 7.30. [a]D = + 128 + 2° (c=0.1; EtOH). v{£i 3420, 2940 (OH), 1740 (CO), 1610, 1290 (Ar) cm - 1 . . 2,4-Dihydroestradiol (XVIII). Prepared from XVII by sodium borohydride re-duction. M.p.: 223—25 °C. Anal.: Calc.: C18H2404 C 71.04 H 7.95; Found: C 71.10 H 7.90. [a]D= + 300 + 2°(c = 0.1; EtOH). v™; 3420, 2940 (OH), 1380,1260(Ar) cm"1. The authors wish to acknowledge the assistance of the Analytical Laboratory of the Institute of Organic Chemistry, Attila József University, Szeged, for the analyses, and to express their thanks for the starting material (estrone) to the Phar-maceutical Works GEDEON RICHTER Budapest). References [1] Werbin, H., C. Holoway: J. Biol. Chem. 223, 651 (1965). ]2] Niederl, J. B„ H. J. Vogel: J. Amer. Chem. Soc. 71, 2566 (1949). 13] Könyves, I., A. Olsson: Acta Chem. Scand. 18, 483 (1964). [4] Patton, T. L.: J. Org. Chem. 24, 1795 (1959). [5] Kraychy, S.: J. Amer. Chem. Soc. 81, 1702 (1959). . [6] Layne, D. S., R. H. Common, W. A. Maw, R. M. Fraps: Nature 181, 351 (1958). [7] Axelrod, L. R„ P. N. Rao, J. W. Goldzicher: Arch. Biochem. Biophys. 87, 152 (1960). [8] Yoshizawa, I., J. Fishman: J. Clin. Endocr. 32, 3 (1971). [9] Brown, B. j„ P. H. Jellinck: Biochem. J. 124, 91 (1971). [10] Elce, J.S.: Biochem. J. 126, 1067 (1972).' [11] Lyttle, C. R„ P. H. Jellinck: Biochem. J. 127, 481 (1972). [12] Knuppen, R., O. Haupt, H. Breuer: Biochem. J. 128, 1369 (1972). [13] Gelbke, H. P., R. Knuppen: J. Chromatogr. 71, 465 (1972). [14] Matkovics, B„ S. E. Rajki, D. G. Szőnyi: Steroids Lipids Res. 3, 118 (1972). [15] Kraychy, S„ T. F. Gallagher: J. Biol. Chem. 229, 523 (1957). СТЕРОИДЫ XXIII. СИНТЕЗ 2- И 4-ГИДРОКСИ-, 2,4-ГИДРОКСИ ЭСТРОНА И ЭСТРАДИОЛА К. Ковач, 3. Раконцаи, Б. Маткович Синтетизированы, через соответствующие нитросоединения, все С-2 и С-4 однозамещен-ные гидроксипроизводные эстрона и эстрадиола. Продукты всех ступеней синтеза выделены и характеризованы. 

Steroids 23. : Synthesi s o f 2 - and 4-Hydrox y an d 2,4-Dihydrox y Derivatives of Estrone and Estradiol

http://acta.bibl.u-szeged.hu/23865/1/phys_chem_019_fasc_003_287-290.pdf

Steroids 23. : Synthesi s o f 2 - and 4-Hydrox y an d 2,4-Dihydrox y Derivatives of Estrone and Estradiol

Abstract

Similar works

Full text

Available Versions

University of Szeged