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EPOXY STEROIDS

20240398836 · 2024-12-05

    Inventors

    Cpc classification

    International classification

    Abstract

    A drug on the basis of 7,19-epoxy steroids, comprising a compound of the general formula (I)

    ##STR00001## or a physiologically acceptable salt thereof, wherein R.sup.1=H; methyl, aryl-CH.sub.2, R.sup.xC(O) or R.sup.xOC(O), or R.sup.1O is dispensed with if there is a double bond between 4-5, W.sup.1, W.sup.2, independently of each other, represent H, OH, CHR.sup.2OH, CHR.sup.2R.sup.x, CR.sup.2R.sup.xOH or W.sup.1 and W.sup.2 together represent O, R.sup.2=saturated or unsaturated, optionally branched alkyl groups with 4 to 9 C atoms, wherein these alkyl groups can also have a cyclopropane or cyclobutane substructure; R.sup.3=methyl or ethyl, X.sup.1, X.sup.2 independently of each other represent R.sup.x or OR.sup.5, or together represent O, NOR.sup.4; Y.sup.1, Y.sup.2 independently of each other represent R.sup.x or OR.sup.5, or together represent O, NOR.sup.4; or X.sup.1 and Y.sup.1 together form a double bond or an epoxy group; Z.sup.1, Z.sup.2 independently of each other represent R.sup.x or OR.sup.5, or together represent O, NOR.sup.4, CH.sub.2; R.sup.4 independently of each other represent H, alkyl or aryl; and R.sup.5 independently of each other represent H; alkyl, aryl-CH.sub.2, R.sup.xC(O) or R.sup.xOC(O), SO.sub.3H, SO.sub.3CH.sub.3, PO(OH).sub.2 or glycosyl; with each R.sup.x independently of each other representing H, saturated or unsaturated, optionally branched alkyl groups, saturated or unsaturated, optionally branched substituted alkyl, aryl, heteroaryl, PEG, SO.sub.3H, PO(OH).sub.2, or glycosyl; and wherein up to 3 hydrogens can be substituted by fluorine.

    Claims

    1. Medicaments based on 7,19-epoxy-steroids comprising a compound of general formula I, ##STR00035## or a physiologically tolerable salt thereof, wherein in the formula: R.sup.1=H; methyl, aryl-CH.sub.2, R.sup.xC(O), or R.sup.xOC(O); or R.sup.1O is omitted if a double bond exists between 4-5; W.sup.1, W.sup.2 are independently H, OH, CHR.sup.2OH, CHR.sup.2R.sup.x, CR.sup.2R.sup.xOH, or W.sup.1 and W.sup.2 are together O, R.sup.2=saturated or unsaturated, optionally branched, alkyl radicals with 4 to 9 carbon atoms, wherein such alkyl radicals may also have a cyclopropane or cyclobutane substructure; R.sup.3=methyl or ethyl, X.sup.1, X.sup.2 are independently R.sup.x or OR.sup.5, or together form O, NOR.sup.4; Y.sup.1, Y.sup.2 are independently R.sup.x or OR.sup.5, or together form O, NOR.sup.4; or X.sup.1 and Y.sup.1 together form a double bond or epoxy group; Z.sup.1, Z.sup.2 are independently R.sup.x or OR.sup.5, or together form O, NOR.sup.4, CH.sub.2; R.sup.4 are independently H, alkyl or aryl; and R.sup.5 are independently H; alkyl, aryl-CH.sub.2, R.sup.xC(O) or R.sup.xOC(O), SO.sub.3H, SO.sub.3CH.sub.3, PO(OH).sub.2 or glycosyl; wherein each R.sup.x is independently H, saturated or unsaturated, optionally branched, alkyl, saturated or unsaturated, optionally branched, substituted alkyl, aryl, heteroaryl, PEG, SO.sub.3H, PO(OH).sub.2, or glycosyl; and wherein up to 3 hydrogens may be substituted by fluorine.

    2. The medicament of claim 1, wherein formula I has formula A, ##STR00036## and the substituents are as defined in claim 1.

    3. The medicament according to claim 1, wherein (i) R.sup.x has from 1 to 10 carbon atoms, and/or (ii) said substituted alkyl in R.sup.x has OR or C(O)OR as a substituent, and R is H or alkyl or a protective group, or (iii) R.sup.1 is H, Z.sup.1, Z.sup.2 are O, R.sup.3 is methyl.

    4. The medicament according to claim 1, wherein Y.sup.1 is H, Y.sup.2 is OH, or Y.sup.1 is H, Y.sup.2 is OR.sup.5, wherein R.sup.5 is R.sup.xC(O) or R.sup.xOC(O), and R.sup.x is H, alkyl, substituted alkyl, aryl, or heteroaryl.

    5. The medicament according to claim 1, wherein R.sup.x is H, alkyl, and R.sup.2 is 4-methylpentyl.

    6. The medicament according to claim 1, wherein R.sup.1 is H; methyl.

    7. The medicament according to claim 1, (i) wherein R.sup.4 has up to 10 carbon atoms, (ii) wherein R.sup.5 has up to 10 carbon atoms or (iii) wherein R.sup.4 has up to 10 carbon atoms and R.sup.5 has up to 10 carbon atoms.

    8. The medicament according to claim 1, wherein R.sup.x has up to 10 carbon atoms.

    9. The medicament according to claim 1, wherein R.sup.x is alkyl or substituted alkyl.

    10. A process for preparing the compound of formula I of claim 1 comprising step a, followed by conversion of functional groups: ##STR00037## wherein step a is performed under basic conditions.

    11. A method of treating malignant diseases, benign or semimalignant skin diseases comprising the step of administering the compound of formula I of claim 1 to a patient in need thereof.

    12. The method of claim 11, wherein said malignant diseases affect bone marrow or other hematopoietic organs, or are solid tumors or epithelial tumors.

    13. The method of claim 11, wherein said benign or semimalignant skin diseases are psoriasis vulgaris, keloids, or basaliomas.

    14. A method of treating inflammatory and chronic inflammatory diseases comprising the step of administering the compound of formula I of claim 1 to a patient in need thereof.

    15. The method according to claim 11, wherein said treating is combined with a cytostatic.

    16. The method of claim 12 wherein said solid tumors are brain tumors of grade 1-grade 4.

    17. The method of claim 16 wherein said solid tumors are glioblastoma or medulloblastoma.

    18. A method of treating a disease comprising the step of administering the compound of formula I of claim 1 to a patient in need thereof in immunosuppressive therapy.

    19. A method of treating a disease comprising the step of administering the compound of formula I of claim 1 to a patient in need thereof in antiviral, antibacterial, antimycotic, antiprotozoan, or antihelminthic therapy.

    20. The method of claim 15 wherein said cytostatic is selected from the group consisting of cytarabin and vincristine.

    Description

    FIGURES

    [0065] FIG. 1 shows the inhibition of proliferation in Nalm-6 cells.

    [0066] FIG. 2 shows the overcoming of steroid and cytostatic resistances in NaKu cells.

    [0067] FIG. 3 shows the overcoming of polycytostatic resistance in BiBo cells.

    [0068] FIG. 4 shows the overcoming of polycytostatic resistance in 7-CCA cells.

    [0069] FIG. 5 shows synergistic effects with cytarabin and vincristin on Nalm-6 cells.

    [0070] FIG. 6 shows the exclusion of unspecific necrosis in Nalm-6 cells.

    [0071] FIG. 7 shows the selectivity for leukemia cells (Nalm-6 cells) and lymphoma cells (BJAB cells) as compared to human leukocytes.

    [0072] FIG. 8 shows the effect in cytostatics-resistant solid tumors.

    [0073] FIG. 9 shows the effect in brain tumor cells.

    [0074] FIG. 10 shows the effect in primary human glioblastoma cells.

    [0075] FIG. 11 shows the effect in primary human pancreatic carcinoma cells.

    [0076] FIG. 12 shows the distribution of the substances according to the invention in serum.

    [0077] FIG. 13 shows the distribution of the substances according to the invention in the brain.

    [0078] FIG. 14 shows the distribution of the substances according to the invention in the liver.

    EXAMPLES

    [0079] The invention is further illustrated by the following Examples.

    Example 1: Synthesis of 3,5-dihydroxy-7,19-epoxy-cholestan-6-one (9, WIL-071)

    [0080] The synthetic sequence for preparing the novel steroids according to the invention is illustrated for the substance WIL-071 as an example. At first, the C-19 methyl group of cholesteryl acetate (1) is hydroxylated according to a method basically known from the literature, before the 19-hydroxycholesteryl acatate (4) is oxidatively functionalized in the B cycle, and the 7,19-epoxy bridge is closed.

    ##STR00007##

    ##STR00008##

    Steps a-c: Synthesis of 3-acetoxy-cholest-.SUP.5.-en-19-ol (4)

    [0081] 200 g (0.23 mol, 1.0 eq.) of cholesteryl acetate (1) in 1.6 liters of dioxan was admixed with 96 g (0.67 mol, 1.5 eq.) of N-bromoacetamide (NBA) and 320 ml of HClO.sub.4 (0.5 N). After 1 h at 0 C. in the dark and 1 h at room temperature, the reaction was stopped by adding a saturated Na.sub.2SO.sub.3 solution (until the reaction mixture changes its color) and water. After extraction with MTBE, the combined organic phases were washed with saturated NaCl solution and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure. The raw product (2, beige solid), with considerable amounts (>40%) of regio-isomeric bromohydrin, was employed in the following step without further purification. For a detailed analysis, a small amount of the bromohydrin 2 was purified by column chromatography.

    [0082] To a solution of (about 25 g) bromohydrin 2 (+regioisomer) in 1.5 L of cyclohexane were added 27.6 g (85.8 mmol, 1.5 eq.) of diacetoxyiodobenzene (DIB) and 8.76 g (68.6 mmol, 1.2 eq.) of iodine. The reaction mixture was heated under reflux for 1 h under irradiation by a 150 Watt mercury vapor lamp. The purple reaction mixture was cooled to room temperature, and quenched by adding a saturated Na.sub.2SO.sub.3 solution and water (until the color changes). After extraction with EtOAc, the combined organic phases were washed with saturated NaCl solution and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, and the raw product (3, brown viscous oil) could be employed in the next step without further purification. This procedure was performed for a total of seven times. A small amount of 3 was purified by column chromatography for analytical purposes.

    [0083] To a solution of compound 3 (about 50 g) in 1.6 L of i-PrOH were added 37.8 g (0.58 mol, 5.0 eq.) of zinc powder and 94 mL (98 g, 1.64 mol, 14.0 eq.) of acetic acid. The reaction mixture was heated under reflux for 3 h, then cooled to room temperature, and filtered through Celite. The clear yellow solution was concentrated to a volume of 100 mL, and extracted with MTBE after the addition of water. The combined organic phases were washed with saturated NaCl solution and dried over MgSO.sub.4. The solvent was removed under reduced pressure. This procedure was performed for a total of four times. The product 4 was obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 10:1) in the form of a beige solid (74 g, 0.17 mol, 36% over three stages).

    [0084] Melting point: 112 C.-114 C.

    [0085] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=5.76 (d, J=5.0 Hz, 1H), 4.66-4.60 (m, 1H), 3.82 (d, J=11.2 Hz, 1H), 3.61 (dd, J=11.4, 9.1 Hz, 1H), 2.41 (ddd, J=13.0, 4.9 Hz, 2.1 Hz, 1H), 2.28-2.23 (m, 1H), 2.02 (s, 3H), 1.95 (dt, J=13.8, 3.4 Hz, 1H), 1.90-0.93 (m, 26H), 0.90 (d, J=6.5 Hz, 3H), 0.85 (dd, J=6.6, 2.2 Hz, 3H), 0.72 (s, 3H).

    [0086] .sup.13C NMR (125 MHz, CDCl.sub.3): [ppm]=170.6, 134.7, 128.4, 73.6, 62.9, 57.7, 56.2, 50.4, 42.7, 41.7, 40.1, 39.6, 38.4, 36.3, 35.9, 33.5, 33.2, 31.4, 28.4, 28.2, 28.2, 24.2, 24.0, 23.0, 22.7, 21.9, 21.5, 18.8, 12.4.

    [0087] FT-IR (ATR): [cm.sup.1]=3507 (bw), 2944 (s), 2934 (s), 2870 (m), 1734 (m), 1713 (s), 1469 (m), 1444 (m), 1381 (m), 1370 (m), 1254 (s), 1245 (vs), 1029 (vs), 977 (m), 961 (m), 911 (w), 886 (w), 824 (w), 807 (w), 742 (w), 670 (w), 610 (w).

    [0088] HR-MS: (ESI, 70 eV)=m/z calculated for C.sub.29H.sub.48O.sub.3Na.sup.+ [M+Na].sup.+ 467.3496 u, found: 467.3493 u.

    Step d: Synthesis of 3-acetoxy-19-methoxymethyloxy-cholestan-.SUP.5.-ene (5)

    [0089] 42 g (78.7 mmol, 1.0 eq.) of 19-hydroxycholesteryl acetate (4) in 315 mL of 5 CH.sub.2(OMe).sub.2 was admixed with 1.64 g (18.9 mmol, 0.2 eq.) of lithium bromide and 1.8 g (9.44 mmol, 0.1 eq.) of p-toluenesulfonic acid (as monohydrate), and the reaction mixture was stirred at room temperature for 5 h. After complete conversion was observed (TLC control), the reaction was stopped by adding a saturated NaCl solution, and the aqueous phase was extracted with MTBE. The combined organic phases were washed with a saturated NaHCO.sub.3 solution and an NaCl solution, and dried over MgSO.sub.4. The product 5 could be obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 5:1) in the form of a white solid (26.9 g, 55.0 mol, 70%).

    [0090] Melting point: 84 C.-85 C.

    [0091] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=5.60 (d, J=5.4 Hz, 1H), 4.67-4.58 (m, 3H), 3.73 (d, J=10.3 Hz, 1H), 3.48 (d, J=10.3 Hz, 1H), 3.37 (s, 3H), 2.40 (ddd, J=13.0, 5.2, 2.2 Hz, 1H), 2.33 (t, J=12.2 Hz, 1H), 2.11 (dt, J=13.8, 3.7 Hz, 1H), 2.03 (s, 3H), 2.02-0.96 (m, 25H), 0.91 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.3 Hz, 6H), 0.70 (s, 3H).

    [0092] .sup.13C NMR (125 MHz, CDCl.sub.3): [ppm]=170.7, 135.8, 126.2, 97.0, 73.8, 69.1, 57.4, 56.2, 55.6, 50.5, 42.6, 40.5, 40.2, 39.7, 38.5, 36.3, 36.0, 33.4, 32.9, 31.7, 28.4, 28.2, 28.2, 24.3, 24.0, 23.0, 22.7, 22.0, 21.6, 18.8, 12.2.

    [0093] FT-IR (ATR): [cm.sup.1]=3419 (br w), 2932 (m), 2868 (m), 1733 (m), 1498 (w), 1467 (w), 1444 (w), 1366 (m), 1242 (s), 1144 (m), 1112 (m), 1096 (m), 1046 (s), 1033 (s), 986 (m), 962 (m), 943 (m), 915 (m), 881 (w), 845 (w), 824 (w), 814 (w), 727 (w), 703 (w), 679 (w), 630 (w), 607 (m), 580 (w).

    Step e: Synthesis of 3-acetoxy-5-hydroxy-19-methoxymethyloxy-cholestan-6-one (6)

    [0094] 32.4 g (66.3 mmol, 1.0 eq.) of compound 5 was dissolved in 400 mL of CH.sub.2Cl.sub.2, and admixed with 25.3 g (113 mmol, 1.7 eq.) of mCPBA (70%). The reaction mixture was stirred at room temperature for 1 h (when a complete conversion was reached, a white precipitate was observed, which is not necessarily required, however), and cooled to 0 C. The white precipitate formed was dissolved by adding 950 mL of acetone, and the clear solution was admixed with a solution of 24 g (239 mmol, 3.6 eq.) of CrO.sub.3 in 80 mL of water. After 10 minutes at 0 C., the reaction mixture was taken to room temperature, and stirred for 1 h. Subsequently, the previously described procedure was repeated with 7.82 g (78.2 mmol, 1.18 eq.) of CrO.sub.3 in 35 mL of water. After 2 h, the reaction mixture was quenched by adding a saturated NaHCO.sub.3 solution, and the aqueous phase was extracted with EtOAc. The combined organic phases were washed with water, a saturated NaHCO.sub.3 solution and a saturated NaCl solution. The clear yellow solution was dried over MgSO.sub.4, and the solvent was removed under reduced pressure. Subsequently, the desired product 6 was obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 8:1) in the form of a white foam (22.3 g, 42.8 mol, 65%).

    [0095] Melting point: 143 C.-144 C.

    [0096] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=5.11 (m, 1H), 4.50 (s, 2H), 3.64 (d, J=10.6 Hz, 1H), 3.57 (d, J=10.6 Hz, 1H), 3.34 (s, 3H), 2.75 (s, 1H), 2.57 (dd, J=14.6, 11.6 Hz, 1H), 2.20-2.13 (m, 2H), 2.09-2.03 (m, 2H), 2.02 (s, 3H), 1.96-1.82 (m, 3H), 1.76-1.63 (m, 3H, H-1), 1.59-1.43 (m, 5H), 1.39-0.96 (m, 12H), 0.91 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.1 Hz, 6H), 0.68 (s, 3H).

    [0097] .sup.13C NMR (125 MHz, CDCl.sub.3): [ppm]=209.6, 171.1, 97.1, 77.8, 70.4, 67.1, 57.0, 56.3, 56.1, 45.7, 44.1, 43.2, 41.5, 40.0, 39.6, 37.2, 36.3, 35.9, 32.5, 28.3, 28.1, 26.6, 26.1, 24.0, 22.9, 22.7, 21.9, 21.5, 18.8, 12.3.

    [0098] FT-IR (ATR): [cm.sup.1]=3384 (br w), 2939 (m), 2869 (m), 1730 (m), 1713 (s), 1467 (w), 1401 (w), 1382 (m), 1365 (m), 1236 (s), 1150 (m), 1106 (m), 1034 (s), 1012 (s), 967 (m), 940 (m), 920 (m), 904 (m), 871 (w), 834 (w), 734 (w), 664 (w), 941 (w), 609 (w), 553 (w).

    Step f: Synthese von 3-acetoxy-7-bromo-5,6-dihydroxy-6,19-epoxy-cholestan (7)

    [0099] To a solution of 22.3 g (42.8 mmol, 1.0 eq.) of compound 6 in 400 mL of acetic acid was added 6.6 mL (20.5 g, 128 mmol, 3.5 eq.) of bromine in acetic acid (65 mL) and a few drops of HBr (48% aqu.). The reaction mixture was heated at 60 C. for 5 h. Subsequently, the reaction mixture was cooled to room temperature, and quenched with a saturated Na.sub.2SO.sub.3 solution. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with water and a saturated NaCl solution, and dried over MgSO.sub.4. Then, the solvent was removed under reduced pressure, and the product 7 was obtained after purification by column chromatography (c-hex/EtOAc, 6:1) in the form of a white solid (13.4 g, 24.2 mol, 57%).

    [0100] Melting point: 124 C.-125 C.

    [0101] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=5.05-5.00 (m, 1H), 4.12 (d, J=4.9 Hz, 1H), 4.02 (d, J=9.0 Hz, 1H), 3.73 (d, J=9.0 Hz, 1H), 3.09 (s, 1H), 2.89 (s, 1H), 2.19 (ddd, J=12.9, 4.5, 2.2 Hz, 1H), 2.04 (s, 3H), 2.05-1.99 (m, 1H), 1.97-1.83 (m, 5H), 1.74 (t, J=12.4 Hz, 1H), 1.62 (s, 2H), 1.59-0.98 (m, 19H), 0.91 (d, J=6.5 Hz, 3H), 0.87 (dd, J=6.6, 3.0 Hz, 6H), 0.74 (s, 3H).

    [0102] .sup.13C NMR (125 MHz, CDCl.sub.3): [ppm]=170.7, 101.7, 79.4, 69.9, 66.8, 59.4, 55.7, 52.9, 45.4, 43.5, 39.6, 39.0, 38.7, 38.6, 36.2, 35.8, 35.0, 28.2, 28.1, 27.2, 24.0, 24.0, 23.3, 23.0, 22.7, 21.6, 21.5, 18.8, 13.1.

    [0103] FT-IR (ATR): [cm.sup.1]=3418 (br w), 2933 (m), 2868 (m), 1726 (s), 1714 (s), 1498 (w), 1466 (w), 1382 (m), 1366 (m), 1244 (s), 1154 (m), 1131 (m), 1096 (m), 1042 (s), 984 (m), 965 (s), 942 (s), 906 (m), 846 (m), 814 (w), 727 (w), 703 (m), 680 (m), 666 (m), 629 (m), 609 (m), 583 (m).

    Step g: Synthesis of 3-acetoxy-5-hydroxy-7,19-epoxy-cholestan-6-one (8) (WIL-232)

    [0104] 22.6 g (40.7 mmol, 1.0 eq.) of compound 7 was dissolved in 1.8 L of dimethylformamide, and admixed with 38.3 g (0.52 mol, 12.7 eq.) Li.sub.2CO.sub.3 and 16.3 g (0.19 mol, 4.6 eq.) of LiBr. The reaction mixture was stirred at 100 C. for 2 h. Subsequently, the reaction mixture was cooled to room temperature, and diluted with water. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with water and a saturated NaCl solution, and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, and the product 8 was obtained after purification by column chromatography (c-hex/EtOAc, 6:1) in the form of a white solid (17.5 g, 37.0 mol, 91%).

    [0105] Melting point: 146 C.-147 C.

    [0106] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=5.25-5.18 (m, 1H), 4.14 (dd, J=10.2, 1.7 Hz, 1H), 3.91 (d, J=10.2 Hz, 1H), 3.82 (d, J=1.7 Hz, 1H), 2.61 (s, 1H), 2.12-2.03 (m, 3H), 2.03 (s, 3H), 2.00-1.94 (m, 2H), 1.86 (dt, J=13.0, 9.4, 5.8 Hz, 1H), 1.73 (td, J=13.8, 4.7 Hz, 1H), 1.65-1.48 (m, 5H), 1.42-1.04 (m, 13H), 0.98 (dt, J=10.2, 8.7 Hz, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.4 Hz, 6H), 0.74 (s, 3H).

    [0107] .sup.13C NMR (125 MHz, CDCl.sub.3): [ppm]=212.9, 170.5, 78.7, 76.2, 69.2, 63.6, 55.7, 52.0, 45.5, 45.1, 43.2, 40.5, 40.2, 39.6, 36.2, 35.8, 35.2, 28.6, 28.1, 26.2, 23.9, 22.9, 22.9, 22.7, 21.7, 21.5, 21.3, 18.7, 13.1.

    [0108] FT-IR (ATR): [cm.sup.1]=3507 (br w), 2944 (s), 2934 (s), 2870 (m), 1734 (m), 1713 (s), 1469 (m), 1444 (m), 1381 (m), 1370 (m), 1254 (s), 1245 (vs), 1029 (vs), 977 (m), 961 (m), 911 (w), 886 (w), 824 (w), 807 (w), 742 (w), 670 (w), 610 (w).

    [0109] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.29H.sub.46O.sub.5Na.sup.+ [M+Na].sup.+ 497.32375 u, found: 497.32341 u.

    Step h: Synthesis of 3,5-dihydroxy-7,19-epoxy-cholestan-6-one (9) (WIL-071)

    [0110] To a solution of 12.7 g (26.7 mmol, 1.0 eq.) of 8 in 890 mL of methanol was added 5.5 g (40.1 mmol, 1.5 eq.) of K.sub.2CO.sub.3, and the reaction mixture was stirred at room temperature for 1 h. After the reaction was complete (TLC), the reaction mixture was diluted with water. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with NaCl solution, and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, and the residue was purified by column chromatography (c-hex/EtOAc, 1:1). The product 9 (WIL-071) was obtained in the form of a white solid (11.3 g, 26.1 mol, 98%), and subsequently recrystallized from about 400 mL (EtOH/H.sub.2O, 1:1) (9.7 g of highly pure substance).

    [0111] Melting point: 178 C.-180 C.

    [0112] .sup.1H NMR (600 MHz, CDCl.sub.3): [ppm]=4.19-4.13 (m, 2H), 3.88 (d, J=10.2 Hz, 1H), 3.81 (s, 1H), 2.71 (s, 1H), 2.11-2.00 (m, 3H), 1.96-1.82 (m, 3H), 1.75 (s, 1H), 1.65-0.94 (m, 20H), 0.88 (d, J=6.5 Hz, 3H), 0.85 (dd, J=6.6, 2.9 Hz, 6H), 0.73 (s, 3H).

    [0113] .sup.13C NMR (150 MHz, CDCl.sub.3): [ppm]=213.6, 79.2, 76.2, 66.2, 63.7, 55.7, 52.1, 45.5, 45.2, 43.3, 40.6, 40.3, 39.6, 38.8, 36.2, 35.8, 30.2, 28.6, 28.1, 23.9, 22.9, 22.9, 22.7, 21.8, 21.5, 18.7, 13.1.

    [0114] FT-IR (ATR): [cm.sup.1]=3463 (br w), 2948 (s), 2868 (m), 1733 (vs), 1497 (w), 1466 (m), 1414 (w), 1382 (m), 1365 (w), 1340 (w), 1296 (w), 1248 (m), 1225 (w), 1153 (m), 1051 (vs), 968 (m), 958 (m), 887 (w), 818 (w), 779 (w), 756 (m), 713 (w), 637 (w), 558 (m), 539 (w).

    [0115] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.44O.sub.4Na.sup.+ [M+Na].sup.+ 455.31318 u, found: 455.31365 u.

    Synthesis of 5-hydroxy-7,19-epoxy-cholestane-3,6-dione (10) (WIL-241)

    ##STR00009##

    [0116] To a solution of 400 mg (0.93 mmol, 1.0 eq.) of alcohol 9 in 11.5 mL of dichloromethane at 0 C. was added 588 mg (1.39 mmol, 1.5 eq.) of Dess-Martin periodinane, and the reaction mixture was stirred at room temperature for 1 h. After the reaction was complete (TLC), the reaction mixture was diluted with aqueous NaHCO.sub.3 solution and aqueous Na.sub.2S.sub.2O.sub.3 solution. The aqueous phase was extracted with CH.sub.2Cl.sub.2, the combined organic phases were dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (chex/EtOAc, 5:1), and the product 10 was obtained in the form of a white solid (148 mg, 0.34 mmol, 37%).

    [0117] Melting point: 204 C.-205 C.

    [0118] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=4.12 (q, J=10.5 Hz, 2H), 3.93 (d, J=1.6 Hz, 1H), 2.99 (d, J=15.6 Hz, 1H), 2.92-2.87 (m, 1H), 2.48 (m, J=16.3, 8.0 Hz, 2H), 2.43-2.34 (m, 1H), 2.19 (t, J=10.6 Hz, 1H), 2.11-2.00 (m, 2H), 1.93-1.84 (m, 1H), 1.65-1.56 (m, 3H), 1.55-1.06 (m, 14H), 1.03-0.95 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.77 (s, 3H).

    [0119] .sup.13C NMR (126 MHz, CDCl.sub.3): [ppm]=211.5, 206.5, 80.0, 76.8, 63.6, 55.7, 51.9, 47.1, 45.4, 45.1, 43.4, 40.9, 40.2, 39.6, 36.9, 36.2, 35.8, 28.6, 28.1, 23.9, 22.9, 22.9, 22.7, 22.4, 21.8, 18.7, 13.1.

    [0120] FT-IR (ATR): [cm.sup.1]=3459 (bw), 2952 (m), 2940 (m), 2918 (m), 2859 (m), 1742 (s), 1714 (s), 1466 (m), 1382 (m), 1345 (w), 1298 (w), 1252 (w), 1238 (m), 1195 (w), 1163 (m), 1145 (m), 1102 (w), 1079 (w), 1051 (s), 980 (w), 948 (m), 913 (w), 903 (w), 891 (w), 854 (w), 822 (w), 797 (w), 752 (m), 723 (m), 663 (w), 631 (m), 594 (w), 559 (m), 525 (m).

    [0121] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.43O.sub.4.sup.+ [M+H].sup.+ 431.31559 u, found: 431.31544 u;

    [0122] m/z calculated for: C.sub.27H.sub.42O.sub.4Na.sup.+ [M+Na].sup.+ 453.29753 u, found: 453.29761 u.

    Synthesis of 7,19-epoxy-cholest-4-en-3,6-dione (11) (WIL-270)

    ##STR00010##

    [0123] To a solution of 50 mg (0.12 mmol, 1.0 eq.) of 10 in 1.2 mL of toluene was added 1.8 mg (5.8 mmol, 0.05 eq.) of pTsOH-H.sub.2O at room temperature. The reaction mixture was heated at 70 C. and stirred at 70 C. for 7 h. After the reaction was complete (TLC), the reaction mixture was diluted with CH.sub.2Cl.sub.2 and diluted aqueous NaHCO.sub.3 solution. The aqueous phase was extracted with CH.sub.2Cl.sub.2, the combined organic phases were dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (chex/EtOAc, 6:1). The product 11 was obtained in the form of a white solid (44 mg, 0.11 mmol, 92%).

    [0124] Melting point: 127 C.-128 C.

    [0125] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=6.42 (s, 1H), 4.03 (d, J=8.9 Hz, 1H), 3.95 (s, 1H), 3.91 (d, J=8.9 Hz, 1H), 2.50-2.43 (m, 2H), 2.23 (t, J=10.9 Hz, 1H), 2.13 (dt, J=13.2, 3.1 Hz, 1H), 2.06-1.96 (m, 1H), 1.95-1.83 (m, 2H), 1.70-1.62 (m, 1H), 1.62-1.47 (m, 3H), 1.44-1.06 (m, 12H), 1.03-0.96 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.4 Hz, 6H), 0.78 (s, 3H)

    [0126] .sup.13C NMR (126 MHz, CDCl.sub.3): [ppm]=198.6, 197.7, 156.4, 122.7, 77.0, 64.3, 55.7, 52.4, 49.3, 45.4, 43.5, 40.3, 39.6, 38.4, 36.2, 35.8, 34.7, 28.7, 28.1, 24.2, 23.9, 22.9, 22.9, 22.7, 21.9, 18.7, 13.1.

    [0127] FT-IR (ATR): [cm.sup.1]=2954 (m), 2933 (m), 2857 (m), 1724 (m), 1688 (s), 1632 (w), 1465 (m), 1453 (m), 1383 (w), 1366 (w), 1328 (w), 1274 (w), 1245 (w), 1231 (m), 1185 (m), 1113 (w), 1074 (w), 1035 (s), 974 (w), 932 (w), 904 (s), 879 (w), 857 (w), 828 (m), 811 (w), 780 (m), 766 (w), 720 (w), 653 (w), 634 (w), 571 (w), 548 (m), 529 (m), 507 (w).

    [0128] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.41O.sub.3.sup.+ [M+H].sup.+ 413.30502 u, found: 413.30513 u;

    [0129] m/z calculated for: C.sub.27H.sub.40O.sub.3Na.sup.+ [M+Na].sup.+ 435.28697 u, found: 435.28720 u.

    Synthesis of 3,5-dibenzyloxy-7,19-epoxy-cholestan-6-one (12) (WIL 242a) and of 5-benzyloxy-3-hydroxy-7,19-epoxy-cholestan-6-one (13) (WIL 242b)

    ##STR00011##

    [0130] To a solution of 39 mg (0.090 mmol, 1.0 eq.) of 9 in 0.92 mL of dimethylformamide were added 35 mg (0.185 mmol, 2.0 eq.) of CsOH.Math.H.sub.2O, 16.2 mL (0.139 mmol, 1.5 eq.) of BnBr and 4 mg (0.011 mmol, 0.1 eq.) of tetrabutylammonium iodide at room temperature. The reaction mixture was stirred at room temperature for 2 h. After the reaction was complete (TLC), the reaction mixture was diluted with MTBE and diluted aqueous NH.sub.4Cl solution. The aqueous phase was extracted with MTBE, the combined organic phases were washed first with water, then with saturated NaCl solution, subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (chex/EtOAc, 9:1 to 3:1). The product 12 (doubly Bn) was obtained in the form of a colorless oil (22 mg, 0.036 mmol, 40%). In addition, the product 13 (singly benzylated) was also obtained in the form of a colorless oil (7.9 mg, 0.015 mmol, 17%).

    [0131] Analytics relating to 12:

    [0132] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=7.38-7.25 (m, 5H), 5.15 (d, J=11.9 Hz, 1H), 4.79 (d, J=12.0 Hz, 1H), 4.13 (d, J=10.3 Hz, 1H), 3.88 (d, J=10.2 Hz, 2H), 3.67 (s, 1H), 2.65 (dd, J=14.2, 3.5 Hz, 1H), 2.09-1.97 (m, 3H), 1.96-1.73 (m, 4H), 1.63-1.06 (m, 18H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.74 (s, 3H).

    [0133] .sup.13C NMR (126 MHz, CDCl.sub.3): [ppm]=208.4, 139.3, 128.4, 127.1, 80.7, 66.1, 65.5, 63.7, 55.7, 52.6, 45.6, 44.1, 43.0, 41.4, 40.6, 39.6, 36.3, 35.8, 33.2, 30.5, 28.7, 28.2, 23.9, 23.0, 22.9, 22.7, 22.0, 21.9, 18.7, 13.1.

    [0134] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.34H.sub.50O.sub.4Na.sup.+ [M+Na].sup.+545.36013 u, found: 545.36016 u.

    [0135] Analytics relating to 13:

    [0136] .sup.1H NMR (500 MHz, CDCl.sub.3): [ppm]=7.38-7.23 (m, 8H), 7.16 (d, J=6.9 Hz, 2H), 5.07 (d, J=11.6 Hz, 1H), 4.58-4.51 (m, 3H), 4.15 (d, J=9.9 Hz, 1H), 3.86 (d, J=10.1 Hz, 1H), 3.66 (s, 1H), 3.64-3.55 (m, 1H), 2.72 (dd, J=14.2, 3.4 Hz, 1H), 2.08-1.93 (m, 4H), 1.91-1.83 (m, 1H), 1.79 (dd, J=14.1, 11.4 Hz, 1H), 1.72 (td, J=13.8, 4.7 Hz, 1H), 1.62-1.46 (m, 4H), 1.41-1.06 (m, 13H), 1.02-0.94 (m, 1H), 0.89 (d, J=6.4 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.73 (s, 3H).

    [0137] .sup.13C NMR (126 MHz, CDCl.sub.3): [ppm]=208.7, 139.2, 138.7, 128.6, 128.4, 127.8, 127.3, 127.3, 80.6, 72.3, 70.6, 65.5, 63.7, 55.8, 52.6, 45.6, 44.1, 43.1, 41.6, 40.6, 39.6, 36.3, 35.8, 30.2, 28.7, 28.2, 27.3, 23.9, 23.0, 23.0, 22.7, 21.9, 21.8, 18.7, 13.1, 0.2.

    [0138] FT-IR (ATR): [cm.sup.1]=3064 (w), 3031 (w), 2949 (s), 2867 (s), 1733 (s), 1496 (w), 1454 (m), 1382 (w), 1364 (w), 1307 (w), 1262 (w), 1228 (w), 1206 (w), 1179 (w), 1142 (w), 1090 (s), 1054 (m), 1001 (w), 953 (w), 873 (w), 842 (w), 803 (w), 734 (s), 693 (s), 557 (w).

    [0139] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.41H.sub.56O.sub.4Na.sup.+ [M+Na].sup.+635.40708 u, found: 635.40751 u.

    Synthesis of (7,19-epoxy-5-hydroxy-6-keto-cholestan-3-yl) ester of propionic acid (14) [WIL-313]

    ##STR00012##

    [0140] To a solution of 100 mg (0.232 mmol, 1.0 eq.) of alcohol 9 in 0.58 mL of pyridine was added 0.30 ml (2.32 mmol, 10 eq.) propionic anhydride at room temperature. The reaction mixture was stirred at room temperature for 25 h, and after the reaction was complete (TLC), it was diluted with ethyl acetate. The organic phase was washed twice with 1 M hydrochloric acid, and once with water, and subsequently dried over MgSO.sub.4. The solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica (cHex/EtOAc, 10:1). The product 14 could be obtained in the form of a white solid (92 mg, 0.19 mmol, 81%).

    [0141] Analytics relating to 14:

    [0142] Melting point: 50 C.-55 C.

    [0143] Rf: 0.50 (3:1, cHex:EtOAc).

    [0144] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.23 (tt, J=11.1, 5.5 Hz, 1H), 4.19-4.11 (m, 1H), 3.91 (d, J=10.2 Hz, 1H), 3.82 (d, J=1.5 Hz, 1H), 2.61 (s, 1H), 2.29 (q, J=7.5 Hz, 2H), 2.13-2.03 (m, 3H), 2.00-1.93 (m, 2H), 1.91-1.82 (m, 1H), 1.73 (td, J=13.9, 4.7 Hz, 1H), 1.65-1.47 (m, 4H), 1.44-1.07 (m, 17H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.6 Hz, 3H), 0.86 (dd, J=6.7, 2.4 Hz, 6H), 0.74 (s, 3H).

    [0145] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: (126 MHz, CDCl.sub.3): [ppm]=212.89, 173.96, 78.67, 76.18, 68.93, 63.57, 55.73, 52.03, 45.52, 45.09, 43.22, 40.53, 40.24, 39.59, 36.22, 35.77, 35.27, 28.64, 28.13, 28.00, 26.24, 23.88, 22.93, 22.89, 22.69, 21.71, 21.27, 18.74, 13.07, 9.30.

    [0146] FT-IR (ATR): [cm.sup.1]=3444 (bw), 2947 (m), 2868 (m), 1737 (s), 1463 (m), 1381 (m), 1350 (m), 1275 (w), 1261 (w), 1181 (s), 1155 (m), 1082 (m), 1046 (m), 1022 (m), 975 (w), 952 (m), 889 (w), 869 (m), 824 (w), 808 (w), 757 (m), 711 (w), 663 (w), 637 (w), 559 (m).

    [0147] HR-MS: (GC-EI-MS, 70 eV)=m/z calculated for: C.sub.30H.sub.48O.sub.5[M].sup.+ 488.34963 u, found: 488.34896 u.

    [0148] [].sub..sup.20: c=0.555 g/100 mL, CHCl.sub.3: [0149] [].sub.365.sup.20=108.83 (0.15), [0150] [].sub.436.sup.20=64.44 (0.22), [0151] [].sub.546.sup.20=37.48 (0.00), [0152] [].sub.579.sup.20=32.43 (0.15), [0153] [].sub.589.sup.20=30.45 (0.00).

    Synthesis of 7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl-methyl carbonate (15) [WIL-357]

    ##STR00013##

    [0154] To a solution of 99 mg (0.231 mmol, 1.0 eq.) of alcohol 9 in 0.49 mL of dichloromethane were added 0.07 ml (8.8 mmol, 3.8 eq.) of pyridine, 3 mg (0.025 mmol, 0.1 eq.) of DMAP and 0.03 (0.38 mmol, 1.6 eq.) of methyl chloroformate at 0 C. After 1.5 h at 0 C., the reaction mixture was heated at room temperature, and stirred at room temperature for 24 h. After the reaction was complete (TLC), the reaction mixture was diluted with an aqueous saturated NaCl solution and water. The aqueous phase was extracted three times with dichloromethane, and the combined organic phases were washed with 1 M hydrochloric acid, and with saturated aqueous NaCl solution, and subsequently dried over MgSO.sub.4. The solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica (cHex/EtOAc, 10:1). The product 15 could be obtained in the form of a white solid (93 mg, 0.19 mmol, 81%).

    [0155] Analytics relating to 15:

    [0156] Melting point: 64 C.-65 C.

    [0157] Rf: 0.67 (1:1, cHex:EtOAc).

    [0158] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.09 (tt, J=11.1, 5.4 Hz, 1H), 4.12 (dd, J=10.4, 1.6 Hz, 1H), 3.90 (d, J=10.3 Hz, 1H), 3.82 (d, J=1.7 Hz, 1H), 3.77 (s, 3H), 2.63 (s, 1H), 2.17 (ddd, J=13.0, 5.2, 1.6 Hz, 1H), 2.13-1.99 (m, 4H), 1.86 (dtd, J=13.1, 9.5, 5.9 Hz, 1H), 1.73 (td, J=13.7, 4.5 Hz, 1H), 1.66-1.55 (m, 3H), 1.51 (dt, J=13.2, 6.6 Hz, 1H), 1.43-1.07 (m, 14H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.74 (s, 3H).

    [0159] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.72, 155.19, 78.65, 76.12, 73.29, 63.51, 55.73, 54.82, 52.02, 45.52, 45.09, 43.20, 40.51, 40.24, 39.59, 36.22, 35.76, 35.21, 28.64, 28.13, 26.18, 23.88, 22.93, 22.88, 22.69, 21.71, 21.22, 18.74, 13.08.

    [0160] FT-IR (ATR): [cm.sup.1]=3457 (bw), 2950 (m), 2867 (w), 1743 (s), 1443 (m), 1383 (w), 1320 (w), 1269 (s), 1258 (s), 1227 (m), 1171 (w), 1154 (w), 1046 (m), 1018 (m), 952 (m), 935 (m), 877 (w), 825 (w), 792 (m), 757 (w), 721 (w), 664 (w), 625 (w), 559 (w), 530 (w).

    [0161] HR-MS: (GC-EI-MS, 70 eV)=m/z calculated for: C.sub.27H.sub.46O.sub.6[M].sup.+ 490.32889 u, found: 490.32834 u.

    [0162] [].sub..sup.20: c=0.58 g/100 mL, CHCl.sub.3: [0163] [].sub.365.sup.20=105.11 (0.08), [0164] [].sub.436.sup.20=61.21 (0.00), [0165] [].sub.546.sup.20=35.34 (0.00), [0166] [].sub.579.sup.20=30.46 (0.08), [0167] [].sub.589.sup.20=28.79 (0.00),

    Synthesis of N-Boc-glycin (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl) ester (16) [WIL-360]

    ##STR00014##

    [0168] To a solution of 199 mg (0.460 mmol, 1.0 eq.) of alcohol 9 in 4.6 mL of dichloromethane were added 0.12 ml (1.5 mmol, 3.4 eq.) of pyridine, 195 mg (1.02 mmol, 2.2 eq.) of EDC.Math.HCl, 18 mg (0.14 mmol, 0.3 eq.) of DMAP and 89 mg (0.51 mmol, 1.1 eq.) of N-Boc-glycine under an argon atmosphere. The reaction mixture was stirred at room temperature for 3.5 h. After the reaction was complete (TLC), the reaction mixture was diluted with 20 ml of ethyl acetate. The organic phase was washed once each with saturated aqueous NaHCO.sub.3 solution, water and saturated aqueous NaCl solution. The organic phase was subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (10:1, cHex/EtOAc), and the product 16 was obtained in the form of a colorless solid (262 mg, 0.444 mmol, 96%).

    [0169] Analytics relating to 16:

    [0170] Melting point: 76 C.-77 C.

    [0171] Rf: 0.42 (3:1, cHex:EtOAc).

    [0172] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.29 (tt, J=11.1, 5.5 Hz, 1H, H-3), 5.04-4.71 (m, 1H, NH), 4.12 (d, J=10.8 Hz, 1H, H-19), 3.94-3.78 (m, 4H, H-29, H-19, H-7), 2.64 (s, 1H, OH), 2.12-1.95 (m, 5H), 1.91-1.81 (m, 1H), 1.73 (td, J=13.7, 4.6 Hz, 1H), 1.63-1.54 (m, 4H), 1.50 (dt, J=13.3, 6.6 Hz, 1H), 1.45 (s, 9H), 1.41-1.05 (m, 13H), 0.98 (q, J=9.1 Hz, 1H), 0.89 (d, J=6.6 Hz, 3H), 0.85 (dd, J=6.6, 2.4 Hz, 6H), 0.74 (s, 3H).

    [0173] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.73, 169.86, 155.81, 80.13, 78.62, 76.15, 70.50, 63.50, 55.73, 52.01, 45.51, 45.08, 43.20, 42.75, 40.51, 40.23, 39.59, 36.21, 35.76, 35.19, 28.63, 28.46, 28.13, 26.15, 23.88, 22.93, 22.88, 22.69, 21.69, 21.21, 18.74, 13.07.

    [0174] FT-IR (ATR): [cm.sup.1]=3387 (bw), 2951 (m), 2869 (m), 1740 (s), 1720 (s), 1513 (m), 1467 (w), 1384 (m), 1366 (m), 1282 (w), 1251 (m), 1204 (m), 1169 (s), 1051 (m), 970 (w), 918 (w), 867 (w), 825 (w), 757 (w), 734 (w), 559 (w).

    [0175] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.34H.sub.55NO.sub.7Na [M+Na].sup.+612.38707 u, found: 612.38748 u.

    [0176] [].sub..sup.20: c=0.575 g/100 mL, CHCl.sub.3: [0177] [].sub.365.sup.20=94.90 (0.22), [0178] [].sub.436.sup.20=56.87 (0.00), [0179] [].sub.536.sup.20=33.39 (0.00), [0180] [].sub.579.sup.20=28.98 (0.08), [0181] [].sub.589.sup.20=27.42 (0.08),

    Synthesis of (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl) ester of (tert-butyldimethylsilanyloxy)acetic acid (17) [WIL-361]

    ##STR00015##

    [0182] To a solution of 198 mg (0.458 mmol, 1.0 eq.) of alcohol 1 in 4.6 mL of dichloromethane were added 0.12 ml (1.5 mmol, 3.4 eq.) of pyridine, 197 mg (1.02 mmol, 2.2 eq.) of EDC.Math.HCl, 18 mg (0.14 mmol, 0.3 eq.) of DMAP and 98 mg (0.51 mmol, 1.1 eq.) of TBS-glycolic acid under an argon atmosphere. The reaction mixture was stirred at room temperature for 6 h. After the reaction was complete (TLC), the reaction mixture was diluted with 20 ml of ethyl acetate. The organic phase was washed once each with saturated aqueous NaHCO.sub.3 solution, water and saturated aqueous NaCl solution. The organic phase was subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (20:1, cHex/EtOAc), and the desired product 17 could be obtained in the form of a colorless solid (165 mg, 0.273 mmol, 59%). In addition, the undesirable by-product 18 could also be obtained as a colorless solid (53 mg, 0.080 mmol, 17%).

    [0183] Analytics relating to 17:

    [0184] Melting point: 139 C.-140 C.

    [0185] Rf: 0.59 (3:1, cHex:EtOAc).

    [0186] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.29 (dt, J=11.3, 5.6 Hz, 1H), 4.20 (s, 2H), 4.12 (d, J=9.3 Hz, 1H), 3.90 (d, J=10.2 Hz, 1H), 3.81 (d, J=1.5 Hz, 1H), 2.60 (s, 1H), 2.13-1.94 (m, 5H), 1.91-1.81 (m, 1H), 1.73 (td, J=13.9, 4.7 Hz, 1H), 1.64-1.46 (m, 4H), 1.43-1.06 (m, 14H), 1.02-0.95 (m, 1H), 0.91 (s, 9H), 0.89 (d, J=6.6 Hz, 3H), 0.85 (dd, J=6.6, 2.4 Hz, 6H), 0.74 (s, 3H), 0.09 (d, J=1.3 Hz, 6H).

    [0187] .sup.13C NMR: (151 MHz, CDCl.sub.3) [ppm]: 212.74, 171.29, 78.62, 76.15, 69.83, 63.54, 62.08, 55.73, 52.02, 45.52, 45.07, 43.20, 40.52, 40.23, 39.59, 36.21, 35.77, 35.20, 28.64, 28.13, 26.19, 25.88, 23.88, 22.93, 22.89, 22.69, 21.70, 21.24, 18.74, 18.54, 13.07, 5.25, 5.27.

    [0188] FT-IR (ATR): [cm.sup.1]=3494 (w), 2952 (m), 2931 (m), 2896 (w), 2858 (w), 1741 (m), 1495 (w), 1463 (w), 1441 (w), 1383 (w), 1361 (w), 1252 (m), 1221 (m), 1187 (w), 1147 (s), 1085 (w), 1048 (m), 1021 (m), 977 (w), 961 (w), 888 (w), 835 (s), 780 (s), 755 (m), 712 (m), 663 (w), 636 (w), 602 (w), 559 (w), 474 (w), 434 (w).

    [0189] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.35H.sub.60O.sub.6SiNa [M+Na].sup.+627.40514 u, found: 627.40559 u.

    [0190] [].sub..sup.20: c=0.53 g/100 mL, CHCl.sub.3: [0191] [].sub.365.sup.20=97.86 (0.09), [0192] [].sub.436.sup.20=58.55 (0.09), [0193] [].sub.546.sup.20=34.47 (0.18), [0194] [].sub.579.sup.20=30.25 (0.35), [0195] [].sub.589.sup.20=28.30 (0.16),

    [0196] Analytics relating to 18:

    [0197] Melting point: 170 C.-171 C.

    [0198] Rf: 0.48 (3:1, cHex:EtOAc).

    [0199] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.33 (tt, J=11.1, 5.5 Hz, 1H), 4.69-4.60 (m, 2H), 4.38 (s, 2H), 4.14 (dd, J=10.3, 1.8 Hz, 1H), 3.93 (d, J=10.2 Hz, 1H), 3.84 (d, J=1.7 Hz, 1H), 2.63 (s, 1H), 2.15-1.98 (m, 5H), 1.93-1.84 (m, 1H), 1.75 (td, J=13.8, 4.7 Hz, 1H), 1.65-1.49 (m, 4H), 1.44-1.08 (m, 14H), 1.04-0.97 (m, 1H), 0.94 (s, 9H), 0.91 (d, J=6.5 Hz, 3H), 0.88 (dd, J=6.6, 2.4 Hz, 6H), 0.76 (s, 3H), 0.14 (s, 6H).

    [0200] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.63, 171.31, 166.98, 78.59, 76.13, 70.83, 63.49, 61.59, 60.96, 55.73, 52.00, 45.51, 45.08, 43.19, 40.51, 40.22, 39.59, 36.21, 35.76, 35.14, 28.63, 28.13, 26.09, 25.88, 23.88, 22.93, 22.88, 22.69, 21.69, 21.20, 18.74, 18.54, 13.07, 5.31.

    [0201] FT-IR (ATR): [cm.sup.1]=3523 (w), 2950 (m), 2930 (m), 2885 (w), 2858 (w), 1771 (m), 1754 (m), 1739 (m), 1470 (w), 1444 (w), 1389 (w), 1367 (w), 1255 (w), 1223 (w), 1180 (m), 1141 (s), 1040 (m), 1018 (w), 975 (w), 956 (w), 891 (w), 880 (w), 836 (s), 782 (m), 760 (m), 729 (w), 693 (w), 663 (w), 635 (w), 561 (w), 545 (w), 454 (w), 408 (w).

    [0202] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.37H.sub.62O.sub.8SiNa [M+Na].sup.+685.41062 u, found: 685.41025 u.

    Synthesis of (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl) ester of glycolic acid (19) [WIL-364]

    ##STR00016##

    [0203] A solution of 104 mg (0.172 mmol, 1.0 eq.) of the TBSW ether 17 in 1.7 mL of THF was cooled to 0 C., and subsequently admixed with 0.34 ml (0.34 mmol, 2.0 eq.) of TBAF. After 1.5 h, and if the reaction was complete (TLC), the reaction mixture was admixed with a saturated aqueous NaCl solution. The aqueous phase was extracted three times with ethyl acetate, the combined organic phases were dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (3:1, cHex/EtOAc), and the desired product 19 could be obtained in the form of a colorless solid (76 mg, 0.15 mmol, 90%).

    [0204] Analytics relating to 19:

    [0205] Melting point: 164 C.-166 C.

    [0206] Rf: 0.17 (2:1, cHex:EtOAc).

    [0207] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.35 (tt, J=11.2, 5.5 Hz, 1H), 4.14-4.08 (m, 3H), 3.90 (d, J=10.3 Hz, 1H), 3.81 (d, J=1.7 Hz, 1H), 2.79 (s, 1H), 2.47 (t, J=5.4 Hz, 1H), 2.14-2.02 (m, 3H), 2.02-1.96 (m, 2H), 1.85 (dtd, J=13.1, 9.4, 6.0 Hz, 1H), 1.74 (td, J=13.9, 4.7 Hz, 1H), 1.63-1.54 (m, 3H), 1.52-1.46 (m, 1H), 1.41-1.03 (m, 14H), 1.01-0.93 (m, 1H), 0.88 (d, J=6.5 Hz, 3H), 0.84 (dd, J=6.6, 2.9 Hz, 6H), 0.73 (s, 3H).

    [0208] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.75, 172.97, 78.58, 76.17, 70.93, 63.48, 60.88, 55.73, 52.00, 45.52, 45.06, 43.20, 40.51, 40.22, 39.59, 36.22, 35.76, 35.17, 28.63, 28.14, 26.18, 23.88, 22.93, 22.89, 22.69, 21.70, 21.22, 18.74, 13.07.

    [0209] FT-IR (ATR): [cm.sup.1]=3454 (bw), 2949 (m), 1932 (m), 2867 (m), 1738 (s), 1465 (m), 1446 (m), 1383 (m), 1366 (w), 1223 (s), 1208 (s), 1155 (m), 1097 (s), 1087 (s), 1045 (s), 1020 (m), 973 (m), 961 (m), 920 (w), 888 (w), 824 (m), 780 (w), 757 (m), 716 (w), 663 (w), 637 (w), 559 (m), 522 (m), 477 (m), 431 (m).

    [0210] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.29H.sub.46O.sub.6Na [M+Na].sup.+ 513.31866 u, found: 513.31859 u.

    [0211] [].sub..sup.20: c=0.56 g/100 mL, CHCl.sub.3: [0212] [].sub.365.sup.20=95.59 (0.30), [0213] [].sub.436.sup.20=57.98 (0.08), [0214] [].sub.546.sup.20=34.17 (0.22), [0215] [].sub.579.sup.20=30.12 (0.22), [0216] [].sub.589.sup.20=28.21 (0.14),

    Synthesis of (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl) ester of benzoic acid (20) [WIL-362]

    ##STR00017##

    [0217] To a solution of 150 mg (0.347 mmol, 1.0 eq.) of alcohol 1, 6 mg (0.05 mmol, 0.1 eq.) of DMAP, 0.10 ml (0.69 mmol, 2.0 eq.) of triethylamine in 2.3 ml DCM was added 0.06 ml (0.52 mmol, 1.5 eq.) of benzoyl chloride at 0 C. under an argon atmosphere. After 15 min at 0 C., the reaction mixture was stirred at room temperature for another 15 h. After the reaction was complete (TLC), the reaction mixture was diluted with dichloromethane and water, and the aqueous phase was extracted three times with dichloromethane. The combined organic phases were washed with saturated aqueous NaCl solution, dried over MgSO.sub.4, and subsequently, the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (40:1-15:1; cHex/EtOAc), and the desired product 20 could be obtained in the form of a colorless solid (158 mg, 0.294 mmol, 85%).

    [0218] Analytics relating to 20:

    [0219] Melting 151 C.-152 C.

    [0220] point:

    [0221] Rf: 0.55 (3:1, cHex:EtOAc).

    [0222] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 8.07-8.01 (m, 2H), 7.61-7.54 (m, 1H), 7.45 (t, J=7.8 Hz, 2H), 5.51 (tt, J=11.0, 5.5 Hz, 1H), 4.25 (dd, J=10.3, 1.7 Hz, 1H), 3.97 (d, J=10.3 Hz, 1H), 3.87 (d, J=1.7 Hz, 1H), 2.70 (s, 1H), 2.25 (ddd, J=13.0, 5.3, 1.5 Hz, 1H), 2.19-2.07 (m, 4H), 1.95-1.78 (m, 2H), 1.78-1.59 (m, 3H), 1.53 (dt, J=13.1, 6.6 Hz, 1H), 1.48-1.09 (m, 14H), 1.06-0.98 (m, 1H), 0.92 (d, J=6.5 Hz, 3H), 0.88 (dd, J=6.6, 2.4 Hz, 6H), 0.77 (s, 3H).

    [0223] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.86, 166.01, 133.04, 130.61, 129.71, 128.46, 78.74, 76.21, 69.83, 63.62, 55.74, 52.04, 45.54, 45.12, 43.26, 40.61, 40.26, 39.60, 36.23, 35.78, 35.39, 28.66, 28.14, 26.35, 23.89, 22.94, 22.91, 22.70, 21.74, 21.33, 18.75, 13.09.

    [0224] FT-IR (ATR): [cm.sup.1]=3423 (bw), 2949 (m), 2867 (m), 1739 (m), 1717 (m), 1603 (w), 1585 (w), 1493 (w), 1451 (w), 1382 (w), 1316 (w), 1273 (s), 1227 (m), 1175 (w), 1152 (w), 1112 (m), 1069 (m), 1046 (m), 1027 (m), 954 (m), 887 (w), 856 (w), 822 (w), 757 (w), 710 (s), 686 (m), 663 (w), 637 (w), 600 (w), 559 (m), 524 (w).

    [0225] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.34H.sub.49O.sub.5[M+H].sup.+ 537.35745 u, C.sub.34H.sub.48O.sub.5Na [M+Na].sup.+ 559.33940 u, found: 537.35769 u, 559.33938.

    [0226] [].sub..sup.20: c=0.56 g/100 mL, CHCl.sub.3: [0227] [].sub.365.sup.20=201.62 (0.29), [0228] [].sub.436.sup.20=111.59 (0.22), [0229] [].sub.546.sup.20=63.60 (0.44), [0230] [].sub.579.sup.20=55.44 (0.34), [0231] [].sub.589.sup.20=52.73 (0.37),

    Synthesis of 4-(7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl)-1-methyl ester of itaconic acid (21) [WIL-367]

    ##STR00018##

    [0232] To a solution of 73 mg (0.169 mmol, 1.0 eq.) of alcohol 1 in 1.7 mL of dichloromethane were added 0.042 ml (0.54 mmol, 3.2 eq.) of pyridine, 74 mg (0.39 mmol, 2.3 eq.) of EDC.Math.HCl, 8 mg (0.07 mmol, 0.4 eq.) of DMAP and 28 mg (0.19 mmol, 1.1 eq.) of monomethyl ester of itaconic acid under an argon atmosphere. The reaction mixture was stirred at room temperature for 2.5 h. After the reaction was complete (TLC), the reaction mixture was diluted with 10 ml of ethyl acetate. The organic phase was washed once each with saturated aqueous NaHCO.sub.3 solution, water and saturated aqueous NaCl solution. The organic phase was subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (10:1-8:1; cHex/EtOAc), and the product 21 could be obtained in the form of a colorless solid (65 mg, 0.116 mmol, 69%).

    [0233] Analytics relating to 21:

    [0234] Melting point: 132 C.-134 C.

    [0235] Rf: 0.39 (3:1, cHex:EtOAc).

    [0236] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 6.31 (d, J=1.0 Hz, 1H), 5.69 (d, J=1.1 Hz, 1H), 5.25 (tt, J=11.1, 5.4 Hz, 1H), 4.13 (d, J=10.5 Hz, 1H), 3.90 (d, J=10.2 Hz, 1H), 3.81 (d, J=1.7 Hz, 1H), 3.76 (s, 3H), 3.30 (d, J=1.1 Hz, 2H), 2.57 (s, 1H), 2.12-2.02 (m, 3H), 2.00-1.94 (m, 2H), 1.91-1.82 (m, 1H), 1.72 (td, J=13.8, 4.7 Hz, 1H), 1.63-1.47 (m, 4H), 1.42-1.06 (m, 14H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.4 Hz, 6H), 0.74 (s, 3H).

    [0237] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.83, 170.14, 166.77, 133.91, 128.55, 78.65, 76.16, 69.79, 63.54, 55.73, 52.27, 52.02, 45.52, 45.09, 43.22, 40.53, 40.24, 39.59, 38.19, 36.22, 35.77, 35.14, 28.64, 28.14, 26.10, 23.89, 22.94, 22.89, 22.69, 21.70, 21.24, 18.74, 13.07.

    [0238] FT-IR (ATR): [cm.sup.1]=3422 (bw), 2951 (w), 2930 (w), 2866 (w), 1731 (s), 1644 (w), 1457 (w), 1434 (w), 1375 (w), 1333 (m), 1301 (w), 1201 (s), 1178 (s), 1146 (m), 1097 (w), 1040 (m), 1033 (m), 1023 (m), 975 (w), 956 (m), 936 (w), 920 (w), 884 (w), 826 (m), 813 (m), 783 (w), 754 (m), 712 (w), 644 (w), 584 (w), 562 (m), 545 (w), 459 (w).

    [0239] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.34H.sub.50O.sub.7[M+H].sup.+ 559.36293 u, C.sub.33H.sub.50O.sub.7Na [M+Na].sup.+ 581.34488 u, found: 559.36329 u, 581.34505 u.

    [0240] [].sub..sup.20: c=0.52 g/100 mL, CHCl.sub.3: [0241] [].sub.365.sup.20=75.000, [0242] [].sub.436.sup.20=64.81, [0243] [].sub.546.sup.20=38.14, [0244] [].sub.579.sup.20=33.72, [0245] [].sub.589.sup.20=33.79,

    Synthesis of 7,19-epoxy-3,5-hydroxy-cholestan-6-on-oxime (22) [WIL-368]

    ##STR00019##

    [0246] To a solution of 101 mg (0.231 mmol, 1.0 eq.) of alcohol 1 in 2.3 mL of pyridine were added 64 mg (0.93 mmol, 4.0 eq.) of hydroxylamine hydrochloride and a solution of 78 mg (0.93 mmol, 4.0 eq.) of sodium acetate in 0.5 ml of methanol at room temperature. The reaction mixture was stirred at room temperature for 19 h, and after the reaction was complete (TLC), the reaction mixture was diluted with EtOAc and a saturated aqueous NaCl solution. The aqueous phase was washed with a diluted NaCl solution, water, a saturated aqueous NaCl solution, and a CuSO.sub.4 solution. The organic phase was dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was recrystallized from a mixture of methanol, ethyl acetate and water. The product 22 could be obtained as a colorless solid (35 mg, 0.078 mmol, 34%).

    [0247] Analytics relating to 22:

    [0248] Melting point: 266 C.-267 C. (decomposition).

    [0249] Rf: 0.09 (1:1, cHex:EtOAc); 0.45 (5:5:1, cHex:EtOAc:MeOH)

    [0250] .sup.1H NMR: (600 MHz, DMSO-d.sub.6) [ppm]: 11.02 (s, 1H), 4.62 (s, 1H), 4.36 (d, J=5.9 Hz, 1H), 3.84 (d, J=9.5 Hz, 2H), 3.69 (s, 1H), 3.59 (d, J=9.9 Hz, 1H), 1.95 (d, J=12.7 Hz, 1H), 1.82-1.65 (m, 3H), 1.65-1.57 (m, 1H), 1.55-1.43 (m, 3H), 1.42-1.17 (m, 8H), 1.17-0.99 (m, 8H), 0.97-0.90 (m, 1H), 0.85 (d, J=6.5 Hz, 3H), 0.81 (dd, J=6.6, 2.9 Hz, 6H), 0.67 (s, 3H).

    [0251] .sup.13C NMR: (151 MHz, DMSO-d.sub.6) [ppm]: 158.76, 74.65, 71.50, 63.77, 62.83, 55.16, 52.23, 44.83, 44.31, 42.47, 40.22, 38.92, 38.78, 37.95, 35.67, 35.11, 29.88, 28.23, 27.39, 23.15, 22.67, 22.44, 22.41, 21.13, 21.11, 18.47, 12.78.

    [0252] FT-IR (ATR): [cm.sup.1]=3511 (bw), 3290 (bw), 3197 (bw), 3111 (bw), 2954 (m), 2939 (m), 2921 (m), 2867 (m), 1493 (w), 1469 (m), 1444 (w), 1418 (w), 1384 (w), 1375 (w), 1362 (w), 1337 (w), 1307 (w), 1251 (w), 1208 (w), 1181 (w), 1157 (m), 1131 (w), 1098 (w), 1069 (w), 1042 (m), 1018 (m), 986 (m), 944 (s), 906 (m), 894 (w), 873 (w), 838 (w), 811 (m), 789 (m), 733 (m), 712 (w), 666 (w), 605 (w), 593 (w), 578 (w), 562 (m), 488 (w), 462 (w), 443 (w).

    [0253] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.46NO.sub.4 [M+H].sup.+ 448.34214 u, found: 448.34203 u.

    [0254] [].sub..sup.20: c=0.59 g/100 mL, CHCl.sub.3: [0255] [].sub.365.sup.20=141.08 (0.16), [0256] [].sub.436.sup.20=84.58 (0.00), [0257] [].sub.546.sup.20=48.41 (0.08), [0258] [].sub.579.sup.20=41.58 (0.16), [0259] [].sub.589.sup.20=38.03 (0.16),

    Synthesis of (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl)methyl ester of fumaric acid (23) [WIL-369]

    ##STR00020##

    [0260] To a solution of 100 mg (0.23 mmol, 1.0 eq.) of alcohol 1 in 2.3 mL of dichloromethane were added 0.06 ml (0.69 mmol, 3.0 eq.) of pyridine, 97 mg (0.051 mmol, 2.2 eq.) of EDC.Math.HCl, 8 mg (0.07 mmol, 0.3 eq.) of DMAP and 37 mg (0.25 mmol, 1.1 eq.) of monomethyl ester of fumaric acid under an argon atmosphere. The reaction mixture was stirred at room temperature for 2 h. After the reaction was complete (TLC), the reaction mixture was diluted with 20 ml of ethyl acetate. The organic phase was washed once each with saturated aqueous NaHCO.sub.3 solution, water and saturated aqueous NaCl solution. The organic phase was subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (8:1-6:1; cHex/EtOAc), and the product 23 could be obtained in the form of a colorless solid (106 mg, 0.195 mmol, 84%).

    [0261] Analytics relating to 23:

    [0262] Melting point: 153 C.-154 C.

    [0263] Rf: 0.20 (6:1, cHex:EtOAc).

    [0264] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 6.85 (d, J=1.1 Hz, 2H), 5.36 (tt, J=11.1, 5.5 Hz, 1H), 4.17 (dd, J=10.3, 1.7 Hz, 1H), 3.94 (d, J=10.3 Hz, 1H), 3.85 (d, J=1.7 Hz, 1H), 3.83 (s, 3H), 2.67 (s, 1H), 2.18-2.01 (m, 5H), 1.94-1.84 (m, 1H), 1.77 (td, J=13.8, 4.6 Hz, 1H), 1.69-1.57 (m, 3H), 1.53 (dt, J=13.1, 6.6 Hz, 1H), 1.45-1.08 (m, 14H), 1.04-0.97 (m, 1H), 0.91 (d, J=6.5 Hz, 3H), 0.88 (dd, J=6.6, 2.4 Hz, 6H), 0.76 (s, 3H).

    [0265] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.69, 165.54, 164.36, 134.09, 133.44, 78.61, 76.15, 70.51, 63.52, 55.73, 52.46, 52.01, 45.52, 45.08, 43.22, 40.54, 40.23, 39.59, 36.22, 35.76, 35.16, 28.64, 28.13, 26.12, 23.88, 22.93, 22.89, 22.69, 21.70, 21.23, 18.74, 13.07.

    [0266] FT-IR (ATR): [cm.sup.1]=3453 (bw), 2951 (m), 2868 (m), 1722 (s), 1645 (w), 1495 (w), 1437 (w), 1383 (w), 1302 (s), 1260 (m), 1227 (w), 1155 (s), 1100 (w), 1086 (w), 1033 (m), 1021 (m), 977 (m), 910 (w), 889 (w), 825 (w), 775 (w), 757 (w), 734 (w), 676 (w), 593 (w), 559 (w), 529 (w).

    [0267] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.33H.sub.48O.sub.7[M+H].sup.+ 545.34728 u, C.sub.32H.sub.48O.sub.7Na [M+Na].sup.+ 567.32922 u, found: 545.34756 u, 567.32935 u.

    [0268] [].sub..sup.20: c=0.59 g/100 mL, CHCl.sub.3: [0269] [].sub.365.sup.20=141.08 (0.16), [0270] [].sub.436.sup.20=84.58 (0.00), [0271] [].sub.546.sup.20=48.41 (0.08), [0272] [].sub.579.sup.20=41.58 (0.16), [0273] [].sub.589.sup.20=38.03 (0.16),

    Synthesis of (7,19-epoxy-5-hydroxy-6-on-cholestan-3-yl) ester of methanesulfonic acid (24) [TAS-S389]

    ##STR00021##

    [0274] To a solution of 150 mg (0.347 mmol, 1.0 eq.) of alcohol 1 in 8 mL of pyridine was added 138 ml (1.75 mmol, 5.0 eq.) of mesyl chloride, followed by stirring at 10 C. for 5 h. Subsequently, the reaction mixture was quenched by adding water, and the aqueous phase was extracted with ethyl acetate. The combined organic phases were successively washed with water, 1 N HCl, and a saturated aqueous NaCl solution. The organic phase was subsequently dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (2:1, cHex/EtOAc), and the mesitylated product 24 could be obtained in the form of a colorless solid (175 mg, 0.343 mmol, 99%).

    [0275] Analytics relating to 24:

    [0276] Melting point: 173 C.-176 C.

    [0277] Rf: 0.35 (2:1, cHex:EtOAc).

    [0278] .sup.1H NMR: (600 MHz, CDCl.sub.3) [ppm]: 5.12 (dp, J=11.0, 5.7 Hz, 1H), 4.14-4.08 (m, 1H), 3.92 (d, J=10.3 Hz, 1H), 3.84 (d, J=1.7 Hz, 1H), 3.01 (s, 3H), 2.77 (s, 1H), 2.24 (ddd, J=13.0, 5.4, 1.6 Hz, 1H), 2.21-2.07 (m, 3H), 2.09-2.03 (m, 1H), 1.91-1.82 (m, 1H), 1.81-1.70 (m, 2H), 1.62-1.47 (m, 3H), 1.42-1.05 (m, 14H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.9 Hz, 6H), 0.75-0.73 (m, 3H).

    [0279] .sup.13C NMR: (151 MHz, CDCl.sub.3) [ppm]: 212.21, 78.67, 77.47, 76.06, 63.35, 55.71, 51.95, 45.49, 45.03, 43.13, 40.35, 40.18, 39.57, 38.70, 36.50, 36.19, 35.73, 28.61, 28.11, 27.56, 23.86, 22.92, 22.85, 22.67, 21.67, 21.36, 18.72, 13.05.

    [0280] FT-IR [cm.sup.1]=3499 (w), 2947 (w), 2921 (w), 2865 (w), 1733 (m), (ATR): 1496 (w), 1467 (w), 1455 (w), 1417 (w), 1363 (m), 1356 (m), 1334 (m), 1302 (m), 1262 (w), 1229 (w), 1181 (m), 1169 (s), 1151 (m), 1100 (w), 1087 (w), 1045 (m), 1019 (w), 981 (m), 945 (s), 885 (m), 861 (s), 821 (m), 764 (m), 748 (m), 720 (w), 662 (w), 638 (w), 592 (w), 559 (w), 531 (m), 507 (m), 497 (m), 434 (w).

    [0281] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.28H.sub.46O.sub.6SNa [M+Na].sup.+533.290731 u, found: 533.29086 u.

    [0282] [].sub.365.sup.20: c=0.54 g/100 mL, CHCl.sub.3: [0283] [].sub.365.sup.20=131.61 (0.23), [0284] [].sub.436.sup.20=75.81 (0.08), [0285] [].sub.546.sup.20=43.52 (0.09), [0286] [].sub.579.sup.20=38.12 (0.15), [0287] [].sub.589.sup.20=36.28 (0.00),

    Synthesis of 7,19-epoxy-5-hydroxy-cholest-2-en-6-one (25) [TAS-S392]

    ##STR00022##

    [0288] To a solution of 143 mg (0.280 mmol, 1.0 eq.) of mesylate in 5.0 mL of DMF were added 92 mg (1.06 mmol, 3.8 eq.) of LiBr and 99 mg (1.01 mmol, 3.6 eq.) of Li.sub.2CO.sub.3. The reaction mixture was subsequently heated under reflux for 1 h. After the reaction mixture had cooled down to room temperature, the solvent was removed under reduced pressure, the residue was taken up in water and dichloromethane, and the aqueous phase was extracted with DCM. The combined organic phases were dried with MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (15:1, cHex/EtOAc), and the elimination product 25 could be obtained in the form of a colorless solid (75 mg, 0.18 mmol, 65%).

    [0289] Analytics relating to 25:

    [0290] Melting point: 78 C.-80 C.

    [0291] Rf: 0.56 (2:1, cHex:EtOAc).

    [0292] .sup.1H NMR: (600 MHz, CDCl.sub.3) [ppm]: 5.76-5.66 (m, 2H), 3.91-3.80 (m, 3H), 2.76-2.64 (m, 2H), 2.19-2.08 (m, 3H), 2.06 (dt, J=13.1, 3.1 Hz, 1H), 1.87 (dtd, J=13.1, 9.4, 5.9 Hz, 1H), 1.72-1.36 (m, 8H), 1.35-1.05 (m, 9H), 1.02-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.85 (dd, J=6.6, 3.0 Hz, 6H), 0.75 (s, 3H).

    [0293] .sup.13C NMR: (151 MHz, CDCl.sub.3) [ppm]: 214.86, 125.53, 123.65, 77.16, 77.08, 65.09, 55.74, 51.94, 45.48, 45.35, 43.58, 40.36, 40.26, 39.59, 36.21, 35.77, 33.46, 28.65, 28.12, 24.66, 23.88, 22.93, 22.86, 22.68, 21.54, 18.74, 13.09.

    [0294] FT-IR [cm.sup.1]=3568 (bw), 3464 (bw), 3399 (bw), 3023 (w), 2949 (s), (ATR): 2933 (m), 1895 (m), 2865 (m), 1742 (s), 1646 (w), 1615 (w), 1490 (w), 1464 (m), 1425 (m), 1380 (m), 1366 (w), 1336 (w), 1314 (w), 1285 (w), 1257 (w), 1232 (w), 1207 (w), 1178 (m), 1128 (w), 1099 (w), 1077 (m), 1039 (s), 979 (w), 966 (w), 932 (w), 908 (m), 899 (m), 876 (w), 861 (w), 820 (w), 801 (w), 770 (w), 738 (m), 657 (s), 608 (m), 565 (m), 488 (w), 463 (w), 429 (w).

    [0295] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.43O.sub.3[M+H].sup.+ 415.320671 u, C.sub.27H.sub.42O.sub.3Na [M+Na].sup.+ 437.302616 u, found: 415.32086 u, 437.30267 u.

    [0296] [].sub..sup.20: c=0.55 g/100 mL, CHCl.sub.3: [0297] [].sub.365.sup.20=663.93 (0.31), [0298] [].sub.436.sup.20=290.97 (0.08), [0299] [].sub.546.sup.20=146.73 (0.15), [0300] [].sub.579.sup.20=125.88 (0.08), [0301] [].sub.589.sup.20=119.94 (0.22),

    Synthesis of 7,19-epoxy-5-hydroxy-cholestan-6-one (26) [TAS-S398]

    ##STR00023##

    [0302] To a solution of 70 mg (0.17 mmol, 1.0 eq.) of alkene 25 in 4 ml of ethyl acetate was added 20 mg of Pd/C, and the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 16 h. Subsequently, the reaction mixture was filtered through celite, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (5:1, cHex/EtOAc), and the hydrogenation product 26 could be obtained in the form of a colorless solid (60 mg, 0.14 mmol, 85%). Analytics relating to 26:

    [0303] Melting point: 50 C.

    [0304] Rf: 0.58 (3:1, cHex:EtOAc).

    [0305] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 4.15 (dd, J=10.1, 1.6 Hz, 1H), 3.86 (d, J=10.1 Hz, 1H), 3.79 (d, J=1.7 Hz, 1H), 2.43-2.36 (m, 1H), 2.07 (tdd, J=11.0, 10.1, 9.5, 2.4 Hz, 2H), 1.94 (dd, J=13.8, 4.5 Hz, 1H), 1.92-1.75 (m, 2H), 1.64-1.55 (m, 6H), 1.51 (ddt, J=16.5, 6.6, 3.2 Hz, 2H), 1.44-1.06 (m, 14H), 1.03-0.95 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.74 (s, 3H).

    [0306] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 215.48, 77.89, 76.46, 63.78, 55.75, 52.13, 45.57, 45.05, 44.07, 40.83, 40.33, 39.60, 36.23, 35.78, 30.11, 28.67, 28.13, 23.88, 22.94, 22.91, 22.69, 22.18, 21.43, 20.32, 19.14, 18.75, 13.08.

    [0307] FT-IR [cm.sup.1]=3474 (bw), 2932 (s), 2866 (s), 1734 (s), 1492 (w), (ATR): 1465 (m), 1449 (m), 1382 (m), 1346 (w), 1261 (w), 1237 (w), 1205 (w), 1153 (m), 1132 (w), 1103 (w), 1085 (w), 1043 (s), 1006 (m), 988 (m), 962 (w), 946 (w), 906 (w), 879 (m), 825 (w), 804 (w), 749 (s), 689 (w), 660 (w), 632 (w), 558 (s), 538 (w), 473 (w), 463 (w), 453 (w).

    [0308] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.45O.sub.3[M+H].sup.+ 417.336322 u, C.sub.27H.sub.44O.sub.3Na [M+Na].sup.+ 439.318266 u, found: 417.33657 u, 439.31838 u.

    [0309] [].sub..sup.20: c=0.49 g/100 mL, CHCl.sub.3: [0310] [].sub.365.sup.20=338.16 (0.16), [0311] [].sub.436.sup.20=153.74 (0.10), [0312] [].sub.546.sup.20=80.07 (0.09), [0313] [].sub.579.sup.20=69.18 (0.17), [0314] [].sub.559.sup.20=65.78 (0.10),

    Synthesis of 7,19-epoxy-6-methylen-cholest-3,5-diol (27) [TAS-S399]

    ##STR00024##

    [0315] To a solution of 125 mg (0.35 mmol, 1.5 eq.) of Ph.sub.3PCH.sub.3Br in 2 mL of dry THF was added 39 mg (0.35 mmol, 1.5 eq.) of KO.sup.tBu at 0 C. The reaction mixture was warmed up to room temperature, stirred at room temperature for 30 min, and subsequently cooled down again to 0 C. Subsequently, a solution of 100 mg (0.231 mmol, 1.0 eq.) of alcohol 1 in 3.0 ml of THF was added to the reaction mixture, and subsequently warmed up to room temperature. The reaction was stopped after h by adding water. The aqueous phase was extracted with ethyl acetate, and the combined organic phases were washed with saturated aqueous NaCl solution, dried over MgSO.sub.4, and freed from solvent under reduced pressure. The residue was purified by column chromatography on silica (1:1, cHex/EtOAc), and the alkene 27 could be obtained in the form of a colorless solid (82 mg, 0.19 mmol, 83%).

    [0316] Analytics relating to 27:

    [0317] Melting point: 171 C.-174 C.

    [0318] Rf: 0.20 (1:2, cHex:EtOAc).

    [0319] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.23 (s, 1H), 4.97 (s, 1H), 4.10 (tt, J=11.0, 5.2 Hz, 1H), 3.98 (dd, J=10.0, 1.7 Hz, 1H), 3.91 (s, 1H), 3.71 (d, J=9.9 Hz, 1H), 2.11-2.03 (m, 2H), 1.93-1.80 (m, 4H), 1.69-1.61 (m, 3H), 1.54-1.47 (m, 2H), 1.45-1.06 (m, 16H), 1.02-0.95 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.1 Hz, 6H), 0.75 (s, 3H).

    [0320] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 155.41, 109.49, 75.66, 75.24, 66.82, 63.83, 55.86, 53.00, 45.44, 44.94, 43.90, 43.40, 40.82, 39.62, 39.12, 36.29, 35.76, 30.24, 28.79, 28.15, 23.86, 22.96, 22.94, 22.70, 21.88, 21.71, 18.77, 13.37.

    [0321] FT-IR [cm.sup.1]=3436 (bw), 1934 (s), 1906 (s), 2865 (s), 1657 (w), (ATR): 1489 (w), 1462 (m), 1450 (m), 1437 (m), 1415 (w), 1382 (m), 1365 (m), 1310 (m), 1263 (m), 1222 (m), 1171 (w), 1151 (w), 1122 (w), 1073 (s), 1039 (s), 1029 (s), 1013 (s), 1001 (m), 956 (s), 926 (m), 892 (m), 875 (m), 852 (m), 818 (m), 767 (w), 749 (w), 710 (w), 693 (m), 660 (w), 531 (m), 491 (m), 454 (m).

    [0322] [].sub..sup.20: c=0.57 g/100 mL, CHCl.sub.3: [0323] [].sub.366.sup.20=85.67 (0.30), [0324] [].sub.436.sup.20=56.78 (0.08), [0325] [].sub.546.sup.20=34.45 (0.17), [0326] [].sub.579.sup.20=30.41 (0.17), [0327] [].sub.589.sup.20=29.12 (0.00),

    Synthesis of 3,5-dihydroxy-7,19-epoxy-cholestan-6-one (28) [TAS-S403]

    ##STR00025##

    [0328] To a solution of 121 mg (0.46 mmol, 2.0 eq.) of PPh.sub.3 and 106 mg (0.46 mmol, 2.0 eq.) of DBAD in 2.0 ml of dry THF was added a solution of 100 mg (0.23 mmol, 1.0 eq.) of alcohol 1 in 3.0 ml of dry THF. When the color of the reaction mixture turned from colorless to yellow, 26 ml (0.46 mmol, 2.0 eq.) of acetic acid was added, followed by stirring at room temperature for 16 h. Subsequently, the reaction mixture was adsorbed on silica, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (20:1-6:1; cHex/EtOAc), and the acetate obtained (0.46 mmol, 2.0 eq.) was directly dissolved in 5.0 ml of methanol. To this solution, 34 mg (0.25 mmol, 1.2 eq.) of K.sub.2CO.sub.3 was added, the reaction mixture was stirred at room temperature for 16 h, and subsequently quenched by adding water. The aqueous phase was extracted with ethyl acetate, the combined organic phases were washed with saturated aqueous NaCl solution, dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (2:1, cHex/EtOAc), and the free alcohol 28 could be obtained in the form of a colorless solid (80 mg, 0.18 mmol, 80%).

    [0329] Analytics relating to 28:

    [0330] Melting point: 148 C.-150 C.

    [0331] Rf: 0.11 (2:1, cHex:EtOAc).

    [0332] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 4.18 (p, J=2.9 Hz, 1H), 3.97 (dd, J=10.3, 1.6 Hz, 1H), 3.92 (d, J=10.2 Hz, 1H), 3.81 (d, J=1.6 Hz, 1H), 3.62 (bs, 2H), 2.13-2.04 (m, 4H), 1.92-1.76 (m, 4H), 1.69-1.55 (m, 2H), 1.51 (dt, J=13.1, 6.6 Hz, 1H), 1.47-1.07 (m, 14H), 1.03-0.94 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.74 (s, 3H).

    [0333] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.36, 79.55, 76.56, 66.47, 63.64, 55.73, 51.99, 45.55, 45.08, 43.63, 41.35, 40.24, 39.58, 36.21, 35.77, 35.01, 28.71, 28.63, 28.12, 23.89, 22.93, 22.87, 22.68, 21.46, 18.74, 17.62, 13.06.

    [0334] FT-IR [cm.sup.1]=3428 (bw), 3287 (bw), 2946 (s), 2867 (m), 1735 (s), (ATR): 1459 (m), 1439 (m), 1377 (m), 1344 (w), 1327 (w), 1297 (w), 1244 (m), 1211 (m), 1119 (m), 1089 (m), 1079 (m), 1044 (s), 949 (m), 934 (m), 921 (m), 897 (m), 860 (m), 821 (w), 766 (w), 730 (w), 714 (w), 667 (m), 632 (m), 597 (m), 581 (m), 526 (w), 492 (w), 453 (w).

    [0335] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.45O.sub.4[M+H].sup.+ 433.331236 u, C.sub.27H.sub.44O.sub.4Na [M+Na].sup.+ 455.313181 u, found: 433.33145 u, 455.31325 u.

    [0336] [].sub..sup.20: c=0.53 g/100 mL, CHCl.sub.3: [0337] [].sub.365.sup.20=181.70 (0.71), [0338] [].sub.436.sup.20=82.45 (0.53), [0339] [].sub.546.sup.20=42.89 (0.32), [0340] [].sub.579.sup.20=36.22 (0.26), [0341] [].sub.589.sup.20=32.20 (0.32),

    Synthesis of 3-acetoxy-5-hydroxy-7,19-epoxy-cholestan-6-one (29) [TAS-S405]

    ##STR00026##

    [0342] To a solution of 50 mg (0.116 mmol, 1.0 eq.) of alkohol 28 in 2.0 ml of pyridine was added 110 ml (1.16 mmol, 10 eq.) of Ac.sub.2O. The reaction mixture was stirred at room temperature for 16 h, and then diluted with water. The aqueous phase was extracted with ethyl acetate, the combined organic phases were washed with 1 M HCl and saturated aqueous NaCl solution, and dried over MgSO.sub.4. The solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica (2:1, cHex/EtOAc). The acetate 29 could be obtained in the form of a colorless solid (54 mg, 0.114 mmol, 98%).

    [0343] Analytics relating to 29:

    [0344] Melting point: 122 C.-114 C.

    [0345] Rf: 0.43 (2:1, cHex:EtOAc).

    [0346] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.31-5.26 (m, 1H), 3.94 (s, 2H), 3.77 (d, J=1.6 Hz, 1H), 3.33 (s, 1H), 2.23-2.11 (m, 2H), 2.13-2.03 (m, 5H), 1.92-1.75 (m, 5H), 1.66-1.56 (m, 1H), 1.51 (dt, J=13.1, 6.6 Hz, 1H), 1.46-1.23 (m, 9H), 1.22-1.05 (m, 5H), 1.03-0.95 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.75 (s, 3H).

    [0347] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 210.28, 169.44, 77.16, 76.67, 68.81, 63.38, 55.74, 52.04, 45.57, 44.58, 43.32, 41.07, 40.25, 39.59, 36.22, 35.80, 33.15, 28.67, 28.13, 25.47, 23.92, 22.94, 22.89, 22.69, 21.58, 21.53, 18.75, 18.10, 13.05.

    [0348] FT-IR [cm.sup.1]=3484 (bw), 2948 (m), 2932 (m), 2866 (m), 1737 (s), (ATR): 1465 (w), 1441 (w), 1375 (m), 1248 (m), 1224 (m), 1157 (m), 1119 (w), 1094 (m), 1083 (m), 1043 (s), 969 (w), 914 (w), 893 (w), 852 (w), 832 (m), 769 (w), 749 (m), 713 (w), 657 (w), 631 (w), 606 (w), 563 (m), 524 (w), 494 (w), 427 (w).

    [0349] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.29H.sub.47O.sub.5[M+H].sup.+ 475.241801 u, C.sub.29H.sub.46O.sub.5Na [M+Na].sup.+ 497.323746 u, found: 475.34182 u, 497.32366 u.

    [0350] [].sub..sup.20: c=0.55 g/100 mL, CHCl.sub.3: [0351] [].sub.365.sup.20=87.15 (0.17), [0352] [].sub.436.sup.20=52.85 (0.08), [0353] [].sub.546.sup.20=30.97 (0.22), [0354] [].sub.579.sup.20=27.76 (0.31), [0355] [].sub.589.sup.20=26.12 (0.17),

    Synthesis of 7,19-epoxy-6-methylcholestan-3,5,6-triol (30) [TAS-S406]

    ##STR00027##

    [0356] To a solution of 100 mg (0.23 mmol, 1.0 eq.) of ketone 1 in 5.0 mL of dry diethyl ether was added 0.23 ml (0.69 mmol, 3.0 eq., 3.0 M in Et.sub.2O) of MeMgBr at 78 C. The reaction mixture was slowly warmed up to room temperature, and stirred at room temperature for 2 h. The complete conversion of the starting material was monitored by means of TLC controls, and subsequently, the reaction was stopped by adding a saturated NH.sub.4Cl solution. The aqueous phase was extracted with ethyl acetate, the combined organic phases were washed with saturated aqueous NaCl solution, dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (1:5, cHex/EtOAc), and the alcohol 30 could be obtained in the form of a colorless solid (90 mg, 0.20 mmol, 87%).

    [0357] Analytics relating to 30:

    [0358] Melting point: 218 C.-221 C.

    [0359] Rf: 0.16 (1:5, cHex:EtOAc).

    [0360] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 4.04 (tt, J=10.9, 5.1 Hz, 1H), 3.87 (dd, J=10.0, 1.6 Hz, 1H), 3.63 (d, J=10.0 Hz, 1H), 3.34 (d, J=1.2 Hz, 1H), 2.49 (s, 3H), 2.07-1.98 (m, 2H), 1.90-1.75 (m, 3H), 1.69-1.46 (m, 6H), 1.43-1.23 (m, 11H), 1.21-1.07 (m, 6H), 1.03-0.96 (m, 1H), 0.90 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.3 Hz, 6H), 0.71 (s, 3H).

    [0361] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 76.81, 74.91, 67.92, 64.05, 55.91, 54.37, 45.39, 42.98, 40.80, 40.68, 40.00, 39.64, 39.30, 36.32, 35.85, 30.20, 28.80, 28.15, 24.09, 23.91, 22.95, 22.91, 22.70, 22.63, 22.07, 18.77, 12.69.

    [0362] FT-IR [cm.sup.1]=3449 (bw), 2930 (m), 2847 (m), 1492 (w), 1463 (w), (ATR): 1438 (w), 1378 (m), 1365 (w), 1325 (w), 1279 (w), 1226 (w), 1171 (w), 1125 (w), 1107 (m), 1067 (m), 1043 (s), 982 (w), 960 (w), 935 (m), 920 (w), 881 (w), 852 (w), 819 (w), 757 (w), 646 (m), 523 (m).

    [0363] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.28H.sub.48O.sub.4Na [M+Na].sup.+471.344481 u, found: 471.34455 u.

    [0364] [].sub..sup.20: c=0.54 g/100 mL, CHCl.sub.3: [0365] [].sub.365.sup.20=252.10 (0.17), [0366] [].sub.436.sup.20=157.41 (0.79), [0367] [].sub.546.sup.20=92.28 (0.61), [0368] [].sub.579.sup.20=81.05 (0.08), [0369] [].sub.589.sup.20=77.10 (0.08),

    Synthesis of 5-hydroxy-2,3-7,19-diepoxy-cholestan-6-one (31) [TAS-S407]

    ##STR00028##

    [0370] To a solution of 117 mg (0.28 mmol, 1.0 eq.) of alkene 25 in 5.0 mL of dichloromethane was added 58 mg (0.34 mmol, 1.2 eq.) of mCPBA at 0 C. The reaction mixture was stirred at room temperature for 16 h, and subsequently, the reaction was stopped by adding water. The aqueous phase was extracted with dichloromethane, and the combined organic phases were washed with saturated Na.sub.2SO.sub.3, NaHCO.sub.3 and NaCl solutions. Subsequently, the combined organic phases were dried over MgSO.sub.4, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica (3:1, cHex/EtOAc), and the epoxide 31 could be obtained in the form of a colorless solid (95 mg, 0.22 mmol, 79%).

    [0371] Analytics relating to 31:

    [0372] Melting point: 189 C.-191 C.

    [0373] Rf: 0.20 (2:1, cHex:EtOAc).

    [0374] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 3.96 (d, J=10.1 Hz, 1H), 3.77 (d, J=1.6 Hz, 1H), 3.68 (dd, J=10.1, 1.6 Hz, 1H), 3.52 (s, 1H), 3.46 (dt, J=3.8, 1.9 Hz, 1H), 3.31 (dd, J=5.6, 3.8 Hz, 1H), 2.52-2.40 (m, 2H), 2.10-2.03 (m, 3H), 1.86 (dtd, J=13.2, 9.4, 6.2 Hz, 1H), 1.68-1.56 (m, 3H), 1.50 (dq, J=13.2, 6.6 Hz, 1H), 1.45-1.22 (m, 8H), 1.19-1.04 (m, 5H), 1.01-0.94 (m, 1H), 0.89 (d, J=6.5 Hz, 3H), 0.86 (dd, J=6.6, 2.5 Hz, 6H), 0.74 (s, 3H).

    [0375] .sup.11C NMR: (126 MHz, CDCl.sub.3) [ppm]: 209.29, 77.72, 77.56, 65.18, 55.67, 54.11, 51.71, 51.25, 45.36, 44.75, 43.72, 40.05, 39.95, 39.56, 36.18, 35.79, 29.57, 28.62, 28.11, 23.92, 23.61, 22.93, 22.81, 22.67, 21.27, 18.72, 13.01.

    [0376] FT-IR [cm.sup.1]=3460 (w), 2977 (m), 2943 (m), 2932 (m), 2922 (m), (ATR): 2864 (m), 1746 (s), 1466 (m), 1435 (w), 1382 (w), 1374 (m), 1326 (w), 1304 (w), 1264 (w), 1245 (w), 1234 (w), 1142 (m), 1130 (m), 1096 (m), 1051 (s), 1034 (m), 978 (w), 937 (m), 903 (s), 870 (m), 803 (s), 792 (m), 776 (m), 750 (m), 723 (m), 681 (w), 651 (w), 610 (m), 590 (m), 566 (m), 487 (w), 470 (m), 450 (s), 414 (w).

    [0377] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.27H.sub.43O.sub.4[M+H].sup.+ 431.315586 u, C.sub.27H.sub.42O.sub.4Na [M+Na].sup.+ 453.297531 u, found: 431.31568 u, 453.29764 u.

    [0378] [].sub..sup.20: c=0.602 g/100 mL, CHCl.sub.3: [0379] [].sub.365.sup.20=375.97 (0.08), [0380] [].sub.436.sup.20=161.24 (0.08), [0381] [].sub.546=81.39 (0.13), [0382] [].sub.579.sup.20=70.10 (0.00), [0383] [].sub.589.sup.20=66.61 (0.00),

    Synthesis of 3-acetoxy-19-hydroxy-stigmast-.SUP.5.-ene (33) [WIL-DK04]

    ##STR00029##

    [0384] 7.49 g (16.4 mmol, 1.0 eq.) of sitosteryl acetate (32) in 200 ml of dioxane was admixed with 3.39 g (23.6 mol, 1.5 eq.) of N-bromoacetamide (NBA) and 40 ml of HClO.sub.4 (0.5 N). After 1 h at 0 C. in the dark and 2.5 h at room temperature, the reaction was stopped by adding a saturated Na.sub.2SO.sub.3 solution (until the reaction mixture changes its color) and water. After extraction with MTBE, the combined organic phases were washed with saturated NaCl solution and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure. The raw product (beige solid, 9.06 g), with considerable amounts (>40%) of regio-isomeric bromohydrin, was employed in the following step without further purification.

    [0385] To a solution of the (about 9.06 g) bromohydrin (+regioisomer) from the previous raw product in 500 ml of cyclohexane were added 7.92 g (24.5 mmol, 1.5 eq.) of diacetoxyiodobenzene (DIB) and 4.99 g (19.6 mmol, 1.2 eq.) of iodine. The reaction mixture was heated under reflux for 3 h under irradiation by a 150 Watt mercury vapor lamp. The purple reaction mixture was cooled to room temperature, and quenched by adding a saturated Na.sub.2SO.sub.3 solution and water (until the color changes). After extraction with MTBE, the combined organic phases were washed with water and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, and the raw product (brown viscous oil, 13.6 g) could be employed in the next step without further purification.

    [0386] To a solution of the raw product from the previous reaction (13.6 g) in 400 ml of i-PrOH were added 5.35 g (8.19 mol, 5.0 eq.) of zinc powder and 13.8 mL (98 g, 22.9 mol, 14.0 eq.) of acetic acid. The reaction mixture was heated under reflux for 3 h, then cooled to room temperature, and filtered through Celite. The clear yellow solution was concentrated to a volume of 100 mL, and extracted with MTBE after the addition of water. The combined organic phases were washed with water and a saturated NaCl solution, and subsequently dried over MgSO.sub.4. The solvent was removed under reduced pressure. This procedure was performed for a total of four times. The product 33 was obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 4:1) in the form of a beige solid (2.67 g, 5.82 mol, 35% over three stages).

    [0387] Analytics relating to 33:

    [0388] Melting point: 115 C.-118 C.

    [0389] Rf: 0.33 (4:1, cHex:EtOAc).

    [0390] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.78 (dd, J=4.8, 2.4 Hz, 1H), 4.64 (tt, J=11.4, 4.8 Hz, 1H), 3.83 (d, J=11.4 Hz, 1H), 3.62 (d, J=11.4 Hz, 1H), 2.42 (ddd, J=13.0, 4.9, 2.3 Hz, 1H), 2.31-2.23 (m, 1H), 2.07-1.99 (m, 5H), 1.96 (dt, J=13.9, 3.7 Hz, 1H), 1.90-1.77 (m, 3H), 1.70-1.46 (m, 7H), 1.40-0.98 (m, 11H), 0.95-0.89 (m, 6H), 0.86-0.80 (m, 9H), 0.80-0.76 (m, 1H), 0.73 (s, 3H).

    [0391] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 170.65, 134.66, 128.47, 73.55, 62.87, 57.71, 56.14, 50.45, 45.97, 42.67, 41.75, 40.13, 38.36, 36.28, 34.07, 33.52, 33.24, 31.39, 29.28, 28.39, 28.25, 26.21, 24.23, 23.20, 21.91, 21.54, 19.96, 19.17, 18.91, 12.36, 12.12.

    [0392] FT-IR [cm.sup.1]=3497 (bs), 2956 (s), 2932 (s), 2866 (s), 1728 (m), (ATR): 1464 (s), 1441 (S), 1377 (m), 1255 (m), 1133 (s), 1088 (s), 1033 (m), 960 (s), 916 (s), 883 (s), 841 (s), 810 (s), 798 (s), 623 (s), 585 (s).

    [0393] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.31H.sub.52O.sub.3Na [M+Na].sup.+481.365217 u, found: 481.36525 u.

    [0394] [].sub..sup.20: c=0.95 g/100 mL, CHCl.sub.3: [0395] [].sub.365.sup.20=87.44 (0.44), [0396] [].sub.436.sup.20=53.19 (0.13), [0397] [].sub.546.sup.20=30.46 (0.05), [0398] [].sub.579.sup.20=27.02 (0.05), [0399] [].sub.589.sup.20=26.95 (0.00),

    Synthesis of 3-acetoxy-19-(methoxymethyloxy)stigmastan-.SUP.5.-ene (34) [WIL-DK05]

    ##STR00030##

    [0400] 2.67 g (5.82 mmol, 1.0 eq.) of 19-hydroxystigmastyl acetate (33) in 34 ml of CH.sub.2(OMe).sub.2 was admixed with 300 g (2.31 mmol, 0.6 eq.) of lithium bromide and 0.11 g (0.57 mmol, 0.1 eq.) of p-toluenesulfonic acid monohydrate, and the reaction mixture was stirred at room temperature for 23 h. After complete conversion was observed (TLC control), the reaction was stopped by adding water, and the aqueous phase was extracted with MTBE. The combined organic phases were washed with water and a saturated NaCl solution, and dried over MgSO.sub.4. The product 34 could be obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 10:1) in the form of a brown solid (2.37 g, 4.59 mol, 79%).

    [0401] Analytics relating to 34:

    [0402] Melting point: 83 C.-84 C.

    [0403] Rf: 0.39 (10:1, cHex:EtOAc).

    [0404] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.60 (dt, J=4.4, 1.9 Hz, 1H), 4.69-4.57 (m, 3H), 3.73 (d, J=10.3 Hz, 1H), 3.48 (d, J=10.3 Hz, 1H), 3.37 (s, 3H), 2.40 (ddd, J=13.1, 5.2, 2.2 Hz, 1H), 2.36-2.27 (m, 1H), 2.19-2.15 (m, 1H), 2.11 (dt, J=13.5, 3.5 Hz, 1H), 2.07-1.96 (m, 5H), 1.84 (dtd, J=15.5, 9.2, 6.0 Hz, 2H), 1.79-1.61 (m, 2H), 1.64-1.45 (m, 6H), 1.39-0.98 (m, 15H), 0.97-0.86 (m, 6H), 0.88-0.79 (m, 9H), 0.81-0.75 (m, 1H), 0.70 (s, 3H).

    [0405] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 170.67, 135.81, 126.26, 97.05, 73.83, 69.07, 57.42, 56.14, 55.60, 50.47, 45.99, 42.59, 40.52, 40.18, 38.50, 36.31, 34.09, 33.45, 32.91, 31.73, 29.29, 28.40, 28.19, 26.21, 24.35, 23.21, 21.99, 21.57, 19.96, 19.18, 18.91, 12.19, 12.13.

    [0406] FT-IR [cm.sup.1]=2952 (bs), 2933 (s), 2869 (s), 1728 (m), 1463 (s), (ATR): 1443 (s), 1379 (m), 1369 (s), 1243 (m), 1140 (s), 1112 (s), 1045 (m), 1027 (m), 917 (s), 916 (s), 881 (s), 839 (s), 802 (s), 736 (s), 611 (s), 578 (s).

    [0407] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.33H.sub.56O.sub.4Na [M+Na].sup.+539.407081 u, found: 539.40665 u.

    [0408] [].sub..sup.20: c=0.85 g/100 mL, CHCl.sub.3: [0409] [].sub.365.sup.20=87.44 (0.44), [0410] [].sub.436.sup.20=53.19 (0.13), [0411] [].sub.546.sup.20=30.46 (0.05), [0412] [].sub.579.sup.20=27.02 (0.05), [0413] [].sub.589.sup.20=26.95 (0.00),

    Synthesis of 3-acetoxy-5-hydroxy-19-(methoxymethyloxy)-sitgmastan-6-one (35) [WIL-DK06]

    ##STR00031##

    [0414] 2.27 g (4.39 mmol, 1.0 eq.) of compound 34 was dissolved in 33 ml of CH.sub.2Cl.sub.2, and admixed with 1.28 g (7.47 mmol, 1.7 eq.) of mCPBA (70%). The reaction mixture was stirred at room temperature for 1 h and then cooled down to 0 C. The white precipitate formed was dissolved by adding 81 mL of acetone, and the clear solution was admixed with a solution of 2.37 g (23.4 mmol, 5.4 eq.) of CrO.sub.3 in 8.0 mL of water. After 10 minutes at 0 C., the reaction mixture was taken to room temperature, and stirred for 18 h. Subsequently, the reaction mixture was stopped by adding a saturated NaHCO.sub.3 solution, and the aqueous phase was extracted with EtOAc. The combined organic phases were washed with water and a saturated NaCl solution. The clear yellow solution was dried over MgSO.sub.4, and the solvent was removed under reduced pressure. Subsequently, the desired product 35 was obtained after purification by column chromatography on silica gel (c-hex/EtOAc, 3:1) in the form of a colorless gel (1.51 g, 2.76 mol, 61%).

    [0415] Analytics relating to 35:

    [0416] Rf: 0.19 (3:1, cHex:EtOAc).

    [0417] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.11 (tt, J=11.2, 5.3 Hz, 1H), 4.50 (s, 2H), 3.64 (d, J=10.7 Hz, 1H), 3.57 (d, J=10.6 Hz, 1H), 3.34 (s, 3H), 2.83 (s, 1H), 2.57 (dd, J=14.6, 11.5 Hz, 1H), 2.21-2.12 (m, 2H), 2.08-1.99 (m, 5H), 1.96-1.82 (m, 3H), 1.77-1.63 (m, 3H), 1.59-1.43 (m, 4H), 1.39-0.98 (m, 12H), 0.92 (d, J=6.4 Hz, 4H), 0.88-0.76 (m, 10H), 0.68 (s, 3H).

    [0418] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 209.63, 171.09, 97.12, 77.83, 70.46, 67.13, 57.05, 56.16, 56.05, 45.92, 45.70, 44.09, 43.22, 41.54, 39.99, 37.20, 36.26, 34.01, 32.48, 29.26, 28.31, 26.56, 26.28, 26.09, 24.05, 23.19, 21.93, 21.50, 19.95, 19.16, 18.84, 12.25, 12.11.

    [0419] FT-IR [cm.sup.1]=3417 (bs), 2936 (m), 2870 (s), 1713 (m), 1463 (s), (ATR): 1443 (s), 1379 (m), 1365 (s), 1236 (m), 1150 (s), 1106 (s), 1034 (m), 1012 (s), 967 (s), 941 (s), 919 (s), 835 (s), 666 (s), 608 (s), 581 (s), 552 (s).

    [0420] HR-MS: (ESI, 70 eV)=m/z calculated for: C.sub.33H.sub.56O.sub.6Na [M+Na].sup.+571.396911 u, found: 571.39677 u.

    [0421] [].sub..sup.20: c=0.90 g/100 mL, CHCl.sub.3: [0422] [].sub.365.sup.20=243.15 (0.14), [0423] [].sub.436.sup.20=116.29 (0.14), [0424] [].sub.546.sup.20=60.18 (0.05), [0425] [].sub.579.sup.20=51.96 (0.05), [0426] [].sub.589.sup.20=49.93 (0.10),

    Synthesis of 3-acetoxy-7-bromo-5,6-dihydroxy-6,19-epoxy-stigmastan (36) [WIL-DK07]

    ##STR00032##

    [0427] To a solution of 1.34 g (2.45 mmol, 1.0 eq.) of compound 35 in 25 mL of acetic acid were added 1.37 mL (8.57 mmol, 3.5 eq.) of bromine and 8 drops of HBr (48% aqu.). The reaction mixture was heated at 60 C. for 24 h. Subsequently, the reaction mixture was cooled to room temperature, and quenched with a saturated Na.sub.2SO.sub.3 solution. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with water and saturated NaCl solution, and dried over MgSO.sub.4. Then, the solvent was removed under reduced pressure, and the product 36 was obtained after purification by column chromatography in the form of a white solid (657 mg, 1.13 mmol, 61%).

    [0428] Analytics relating to 36:

    [0429] Melting point: 93 C.-94 C.

    [0430] Rf: 0.21 (5:1, cHex:EtOAc).

    [0431] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.03 (tt, J=11.0, 4.6 Hz, 1H), 4.13 (d, J=4.7 Hz, 1H), 4.06-4.01 (m, 1H), 3.73 (d, J=8.9 Hz, 1H), 2.20 (ddd, J=12.9, 4.4, 2.2 Hz, 1H), 2.10 (s, 1H), 2.07-2.00 (m, 4H), 1.97-1.83 (m, 5H), 1.75 (dd, J=12.9, 11.9 Hz, 1H), 1.70-1.64 (m, 1H), 1.60-1.00 (m, 17H), 0.96-0.89 (m, 4H), 0.88-0.76 (m, 10H), 0.75 (s, 3H).

    [0432] .sup.13C NMR: (151 MHz, CDCl.sub.3) [ppm]: 170.81, 101.79, 79.34, 69.92, 66.86, 59.35, 55.56, 52.87, 45.94, 45.37, 43.44, 38.98, 38.64, 38.53, 36.21, 35.01, 34.01, 29.29, 28.11, 27.18, 26.22, 24.01, 23.28, 23.21, 21.57, 21.54, 19.99, 19.15, 18.85, 13.12, 12.12.

    [0433] FT-IR [cm.sup.1]=3413 (bs), 2954 (m), 2936 (m), 2869 (s), 2050 (s), (ATR): 1713 (m), 1497 (s), 1457 (s), 1377 (m), 1365 (s), 1243 (m), 1153 (s), 1129 (s), 1036 (m), 1095 (s), 985 (s), 945 (s), 906 (s), 844 (s), 702 (s), 677 (s), 628 (s), 608 (s), 528 (s).

    [0434] HR-MS: (GC-EI/MS, 70 eV, 50-250 C.) m/z (%)=81.06980 (88), 109.06468 (100), 255.17394 (55), 396.33803 (30), 414.34872 (45), 474.36932 (10), 502.36426 (<5).

    [0435] [].sub..sup.20: c=1.00 g/100 mL, CHCl.sub.3: [0436] [].sub.365.sup.20=137.92 (3.68), [0437] [].sub.436.sup.20=84.96 (0.12), [0438] [].sub.546.sup.20=50.07 (0.21), [0439] [].sub.579.sup.20=43.78 (0.12), [0440] [].sub.589.sup.20=42.25 (0.28).

    Synthesis of 3-acetoxy-5-hydroxy-7,19-epoxy-stigmastan-6-one (37) [WIL-DK08]

    ##STR00033##

    [0441] 533 mg (0.913 mmol, 1.0 eq.) of compound 36 was dissolved in 46 ml of dimethylformamide and admixed with 858 mg (11.6 mmol, 12.7 eq.) of Li.sub.2CO.sub.3 and 365 mg (4.20 mmol, 4.6 eq.) of LiBr. The reaction mixture was stirred at 100 C. for 2 h. Subsequently, the reaction mixture was cooled to room temperature, and diluted with water. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with water and saturated NaCl solution, and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, and the product 37 was obtained after purification by column chromatography (c-Hex/EtOAc, 2.5:1) in the form of a white solid (321 mg, 0.639 mmol, 54%).

    [0442] Analytics relating to 37:

    [0443] Melting point: 142 C.-143 C.

    [0444] Rf: 0.33 (2.5:1, cHex:EtOAc).

    [0445] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 5.22 (tt, J=11.2, 5.5 Hz, 1H), 4.14 (dd, J=10.3, 1.7 Hz, 1H), 3.91 (d, J=10.2 Hz, 1H), 3.82 (d, J=1.7 Hz, 1H), 2.63 (s, 1H), 2.13-2.01 (m, 6H), 2.00-1.93 (m, 2H), 1.88 (dtd, J=13.0, 9.4, 5.9 Hz, 1H), 1.77-1.49 (m, 5H), 1.42-1.11 (m, 14H), 1.05-0.97 (m, 1H), 0.95-0.87 (m, 4H), 0.87-0.75 (m, 9H), 0.74 (s, 3H).

    [0446] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 212.88, 170.55, 78.65, 76.17, 69.18, 63.55, 55.60, 52.02, 45.94, 45.52, 45.09, 43.21, 40.53, 40.23, 36.13, 35.23, 33.98, 29.24, 28.66, 26.22, 26.11, 23.18, 22.90, 21.70, 21.46, 21.25, 19.93, 19.14, 18.80, 13.07, 12.11.

    [0447] FT-IR [cm.sup.1]=3533 (bs), 2953 (m), 2863 (s), 2112 (s), 1733 (m), (ATR): 1499 (s), 1463 (s), 1375 (m), 1332 (s), 1253 (m), 1151 (s), 1097 (s), 1042 (m), 1024 (s), 963 (s), 945 (s), 910 (s), 892 (s), 870 (s), 753 (s), 710 (s), 655 (s), 635 (s), 606 (s), 574 (s).

    [0448] HR-MS: (GC-EI/MS, 70 eV, 50-250 C.) m/z (%)=81.06982 (100), 109.06469 (100), 255.17406 (65), 396.33804 (35), 414.34868 (50), 474.36956 (10), 502.36451 (<1).

    [0449] [].sub..sup.20: c=0.91 g/100 mL, CHCl.sub.3: [0450] [].sub.365.sup.20=111.64 (0.27), [0451] [].sub.436.sup.20=66.74 (0.16), [0452] [].sub.546.sup.20=39.38 (0.14), [0453] [].sub.579.sup.20=34.36 (0.16), [0454] [].sub.589.sup.20=33.11 (0.14).

    Synthesis of 3,5-dihydroxy-7,19-epoxy-stigmastan-6-one (38) [WIL-DK09]

    ##STR00034##

    [0455] To a solution of 222 mg (0.442 mmol, 1.0 eq.) of 37 in 30 mL of methanol was added 91 mg (0.66 mmol, 1.5 eq.) of K.sub.2CO.sub.3, and the reaction mixture was stirred at room temperature for 1 h. After the reaction was complete (TLC), the reaction mixture was diluted with water. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with NaCl solution, and dried over MgSO.sub.4. Subsequently, the solvent was removed under reduced pressure, the residue was purified by column chromatography (c-Hex/EtOAc, 1:1) any by recrystallization from EtOH/H.sub.2O. The product 38 was obtained in the form of a white solid (110 mg, 0.685 mmol, quant.).

    [0456] Analytics relating to 38:

    [0457] Melting point: 175 C.-176 C.

    [0458] Rf: 0.17 (1:1, cHex:EtOAc).

    [0459] .sup.1H NMR: (500 MHz, CDCl.sub.3) [ppm]: 4.22-4.12 (m, 2H), 3.89 (d, J=10.2 Hz, 1H), 3.82 (d, J=1.7 Hz, 1H), 2.14-2.01 (m, 3H), 1.97-1.84 (m, 3H), 1.71-1.54 (m, 4H), 1.48 (tdd, J=13.4, 11.2, 4.9 Hz, 1H), 1.42-1.11 (m, 14H), 1.05-0.98 (m, 1H), 0.94-0.88 (m, 4H), 0.86-0.75 (m, 10H), 0.74 (s, 3H).

    [0460] .sup.13C NMR: (126 MHz, CDCl.sub.3) [ppm]: 213.65, 79.18, 76.18, 66.23, 63.66, 55.62, 52.06, 45.93, 45.53, 45.19, 43.31, 40.59, 40.28, 38.87, 36.12, 33.99, 30.19, 29.24, 28.67, 26.10, 23.18, 22.91, 21.76, 21.52, 19.93, 19.14, 18.80, 13.07, 12.11.

    [0461] FT-IR [cm.sup.1]=3353 (bs), 2955 (m), 2861 (s), 2050 (s), 1732 (m), (ATR): 1495 (s), 1463 (s), 1376 (m), 1365 (s), 1250 (m), 1225 (s), 1154 (s), 1070 (s), 1049 (m), 969 (s), 914 (s), 88 (s), 872 (s), 853 (s), 818 (s), 754 (s), 710 (s), 6635 (s), 637 (s), 583 (s), 559 (s).

    [0462] HR-MS: (GC-EI/MS, 70 eV, 50-300 C.) m/z (%)=81.06982 (100), 109.06472 (60), 133.10107 (55), 273.18460 (35), 414.34881 (25), 432.35897 (15), 460.35410 (<1).

    [0463] [].sub..sup.20: c=0.86 g/100 mL, CHCl.sub.3: [0464] [].sub.365.sup.20=248.03 (0.20), [0465] [].sub.436.sup.20=120.94 (0.34), [0466] [].sub.546.sup.20=63.82 (0.58), [0467] [].sub.579.sup.20=56.53 (0.54), [0468] [].sub.589.sup.20=53.76 (0.71),

    Example 2: Inhibition of Proliferation in Leukemia Cells (Nalm-6 Cells)

    [0469] In leukemia cells (Nalm-6), cell proliferation is inhibited. After 24 h, 110.sup.5 cells will have proliferated to almost threefold (2.9710.sup.5 cells), while cell proliferation is inhibited up to 100% in a concentration-dependent way by treatment with the steroid WIL 071, see FIG. 1. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    Example 3: Overcoming Steroid and Polycytostatics Resistances in NaKu Cells

    [0470] In steroid- and polycytostatics-resistent leukemia cells (NaKu (ALL)), treatment with the steroid WIL 071 induces as much apoptosis as in the non-resistent leukemia cells (Nalm-6) in a non-concentration-dependent way, see FIG. 2. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    [0471] The steroid WIL 071 overcomes steroid resistence in leukemia cells (NaKu cells). Further, the steroid-resistent leukemia cells (NaKu cells) have co-resistencies for the following cytostatics: [0472] prednisolone, methylprednisolone, dexamethasone, [0473] doxorubicin, daunorubicin, idarubicin, epirubicin [0474] cytarabine, cladribine [0475] etoposide [0476] cyclophosphamide, ifosfamide [0477] vindesine [0478] cisplatin, carboplatin, oxaliplatin

    Example 4: Overcoming of Polycytostatics Resistance in BiBo Cells

    [0479] BiBo cells are lymphoma cells (BJAB cells) that have been made resistent against vincristine, and have a Bcl-2 overexpression as a resistence mechanism. They have co-resistences for the following cytostatics: [0480] vincristine, vinblastine, vindesine, vinorelbin, taxol [0481] cytarabin [0482] daunorubicin.

    [0483] In polycytostatics-resistent lymphoma cells (BiBo cells), treatment with the steroid WIL 071 induces as much apoptosis as in the non-resistent lymphoma cells (BJAB (Burkitt like lymphoma cells)) in a non-concentration-dependent way, see FIG. 3. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments. The steroid WIL 071 overcomes steroid resistence in lymphoma cells (BiBo cells).

    Example 5: Overcoming of Polycytostatics Resistance in 7-CCA Cells

    [0484] 7-CCA cells are lymphoma cells (BJAB cells) that have been made resistent against doxorubin, and have a caspase-3 underexpression as a resistence mechanism. They have co-resistences for the following cytostatics: [0485] vincristine, vinblastine, vindesine, vinorelbin, taxol [0486] cytarabin, fudarabin, cladribin, clofarabin, [0487] daunorubicin, doxorubicin, idarubicin, epirubicin [0488] cisplatin, oxaliplatin

    [0489] In polycytostatics-resistent lymphoma cells (7-CCA cells), treatment with the steroid WIL 071 induces as much apoptosis as in the non-resistent lymphoma cells (BJAB (Burkitt like lymphoma cells)) in a non-concentration-dependent way, see FIG. 4. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments. The steroid WIL 071 overcomes steroid resistence in lymphoma cells (7-CCA cells).

    Example 6: Synergistic Effects with Cytarabin and Vincristine

    [0490] In leukemia cells (Nalm-6 cells), parallel treatment with the steroid WIL 071 and cytarabin or vincristine induces significantly more apoptosis than 1.1 times the sum of the apoptotic rates in treatment by a monotherapy with the corresponding drugs, see FIG. 5. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments. The steroid WIL 071 has synergistic effects with cytarabin and vincristine.

    Example 7: Exclusion of Non-Specific Necrosis

    [0491] What is measured is the release of lactate dehydrogenase (LDH) in leukemia cells (Nalm-6 cells) in the cell supernatant, which is an indication of non-specific cell death (necrosis), because this large protein LDH can cross cell membranes towards the extracellular space only if they are defect, see FIG. 6. Non-specific necrosis can be excluded.

    Example 8: Selectivity

    [0492] In leukemia cells (Nalm-6 cells) and in lymphoma cells (BJAB cells), apoptosis is induced in a concentration-dependent way by treatment with the steroid WIL 071. WIL 071 acts selectively in malignant cells that are proliferating. In contrast, apoptosis is not induced in human primary leukocytes.

    [0493] The DNA fragmentation was measured by flow cytometry, see FIG. 7. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    Example 9: Effect in Cytostatics-Resistant Solid Tumors

    [0494] In contrast to conventional glucocorticoids, the steroid WIL 071 can induce apoptosis also in solid tumor cells. Resistances towards cytostatics are overcome also in neuroblastoma cells (SKNAS cells) (LiOn cellscisplatin-resistant SKNAS cells with an underexpression of caspase-8.

    [0495] The DNA fragmentation was measured by flow cytometry), see FIG. 8. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    Example 10: Effect in Brain Tumor Cells

    [0496] In contrast to conventional glucocorticoids, the steroid WIL 071 can induce apoptosis also in brain tumor cells. The steroid WIL 071 significantly induces apoptosis also in glioblastoma cells (DBTRG05MG cells).

    [0497] The DNA fragmentation was measured by flow cytometry, see FIG. 9. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    Example 11: Effect in Primary Human Glioblastoma Cells

    [0498] In contrast to conventional glucocorticoids, the steroid WIL 071 can induce apoptosis also in primary human brain tumor cells. The steroid WIL 071 significantly induces apoptosis also in primary glioblastoma cells of a patient having a glioblastoma (grade 4).

    [0499] The DNA fragmentation was measured by flow cytometry, see FIG. 10. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    Example 12: Effect on Pancreatic Carcinoma Cells

    [0500] In contrast to conventional glucocorticoids, the steroid WIL 071 can induce apoptosis also in pancreatic carcinoma cells. In a corresponding cell line (DAN-G), the steroid WIL 071 significantly induces apoptosis.

    [0501] The DNA fragmentation was measured by flow cytometry, see FIG. 11. The experiments were performed in triplicate. The error bars show the standard deviations of the measured values of three independent experiments.

    [0502] The standard treatment for pancreatic carcinoma includes 5-fluorouracil. It shows clearly worse apoptosic rates in comparable experiments (Oxaliplatin: 14% apoptosis at 50 M, 5-fluorouracil 24.9% at 30 M, 21.2% at 50 M).

    Example 13: Comparison of the Biological Effects of Different Steroid Derivatives in Leukemia Cells (Nalm-6)

    TABLE-US-00002 Biological effect: half- Overcoming of Steroid maximal apoptosis induction prednisolone derivative (AC.sub.50) [M] resistance [M] WIL-071 17.2 16 WIL-232 25 17.5 WIL-241 41 WIL-242b 37 40 WIL-270 20 18 WIL357 >50 >50 WIL360 >20 20 WIL364 ~18 ~17.5 WIL367 ~8 ~12.5 WIL369 ~3 5 WIL-DK-08 50 ~40 WIL-DK-09 ~15 ~38 TAS-S389 ~20 ~30 TAS-S392 ~46 ~39 TAS-S398 ~80 ~67 TAS-S399 50 ~75 TAS-S403 100 20 TAS-S405 90 ~20 TAS-S406 13 ~17
    Apoptosis Induction in Nalm-6 Cells after Treatment with WIL-071 or Different Steroid Derivatives.

    [0503] The cells were incubated in 6-well plates with 1.0.Math.10.sup.5 cells/mL in two milliliters of medium with the respective steroid derivative in different concentrations. The included solvent control (DMSO) served as a control, which was subtracted from all the other measured values, in order not to take into account the spontaneous apoptosis of the cells that takes place independently of the concentration of active ingredient. After 72 h, a modified cell cycle analysis was performed, in order to quantify the proportion of apoptotic cells by flow cytometry. For each concentration of the steroid, the respective mean values f standard deviations, n=3 (*: p<0.05 vs. DMSO, t test) were determined. Using the curve of the dose-effect relationship (steroid concentrationapoptosis induction) formed from these measured values, the steroid concentration at which apoptosis was triggered in 50% of the tested leukemia cells was read.

    [0504] Prednisolone resistance is said to be overcome if the induction of apoptosis in prednisolone-resistant leukemia cells is as high as it is in normal leukemia cells.

    Example 14: Examination of Tolerability in an Animal Model

    [0505] An incremental increase of dose of the three substances Wil-071, Wil-232 and Wil-369 was performed in 8-week-old female NOG-F (Taconic) mice. Different mice were administered one of up to seven different doses of the test substances. The start was on day 1 with the lowest dose of 10 mg/kg. If no behavioural abnormalities or observable health symptoms showed in the course of the next 24 hours after such injection into the caudal vene of the animal, then the treatment was continued on the next day with the next higher dose. All in all, if possible, the doses 10, 20, 40, 80, 100, 150 and 200 mg/kg are tested. Subsequently, the maximum tolerable dose (MTD) was administered to three more animals, and these were subsequently observed for one week, in order to be able to observe any developing symptoms. For this purpose, the animals were observed and weighed on a daily basis. The body weight is considered an indicator of the general condition of the animals.

    [0506] Following the determination of the MTD, eight animals were again injected the MTD of the substances, and after 1, 2, 8 and 24 hours, 2 animals each were killed. For pharmacokinetics, the liver, brain and serum of the animals were shock-frosted in liquid nitrogen, and analyzed (see Example 15).

    Results

    [0507] All three substances are sparingly soluble in tested injection solutions, so that the maximal dose could not exceed 200 mg/kg.

    [0508] For the MTD experiment, Wil-071 was dissolved in 100% DMSO, and then diluted 1:50 with a 20% (w/v) 2-hydroxypropyl-beta-cyclodextrin (HPCD) solution. Thus, 200 L of injection solution (per mouse) of a 40 mg/kg dose could be achieved. For 80, 100 and 150 mg/kg, the DMSO proportion was increased to up to 4% (v/v) in the final injection solution. In addition, ultrasound had to be employed, and the sample had to be heated at 60 C. for up to 45 min. 200 mg/kg was reached only as a milky suspension. The animals did not show any abnormalities after the injection of the individual doses, and the final section did not show any toxicity macroscopically. However, the injection of the 200 mg/kg suspension resulted in the immediate death of the animal. A section showed signs of pulmonary embolism. Therefore, Wil-071 can be considered non-toxic up to 150 mg/kg. The final MTD could not be determined because of the insufficient solubility.

    [0509] For 100 mg/kg of Wil-232, the end concentration of DMSO in the injection solution was 2%. In order to dissolve Wil-232, the sample also had to be treated with ultrasound, and heated at 60 C. The dose of 150 mg/kg was achieved only with 4% DMSO, and 200 mg/kg was achieved only as a suspension. The injection of the 200 mg/kg dose resulted in the immediate death of the animal (signs of pulmonary embolism in section). Otherwise, there were no signs of toxicity, so that the substance can be considered tolerable up to 150 mg/kg. In this case too, a final MTD cannot be determined because of insufficient solubility.

    [0510] Wil-369 showed the lowest solubility. Using a DMSO stock and 20% HPCD solution, a maximum of only 5 mg/ml was dissolved. A further increase could not be achieved even with more DMSO or HPCD. Thus, above the dose of 40 mg/kg, a higher injection volume was injected in order to reach the corresponding dose. With 100 mg/kg in twice as large injection volumes, the animals lay down on their side for some moments, but recovered quickly. In the further course and in the section, all animals were normal, except for one animal from the 100 mg/kg dose group, which was found dead on day 3 after the injection. For the tolerability under the current solubility conditions, a value of 50 mg/mg/kg is to be applied. This dose was injected into the 8 animals for pharmacokinetics. In this case too, a final MTD cannot be determined because of insufficient solubility.

    Recognition

    [0511] In the concentration range that allows the substances to be dissolved completely, all substances are well tolerable.

    Example 15: Distribution

    [0512] The samples obtained in Example 4 were examined for the contents of the substances. Since WIL-369 can be hydrolyzed to WIL-071, both WIL-369 and WIL-071 were determined in the samples. As far as these substances were detectable in the samples, this is represented in the Figures. The distribution is shown in FIGS. 12 to 14.

    [0513] FIG. 12 shows that WIL-071 is formed when WIL-369 is administered, but the serum levels are cleared quickly; thus, WIL-369 is a prodrug.

    [0514] FIG. 13 shows relevant levels of WIL-071 after administration of WIL-369 or WIL-071 in the brain, which could be effective for more than 8 hours.

    [0515] FIG. 14 shows that a quick degradation takes place in the liver; it is the only place where WIL-369 is found.

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    In General:

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    Cell Cycle Arrest:

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    ER Stress, ROS, Autophagy:

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