CHOLESTANE DERIVATIVES, PREPARATIONS CONTAINING THESE DERIVATIVES AND USE THEREOF

Abstract

The invention relates to brassinosteroid-derived cholestane derivatives, their use in the protection of cell damage, injury and cell death and compositions containing these derivatives. New generation of compounds possess also selective antineurodegenerative properties on neuronal cells and tissues and can be particularly used in the treatment and prophylaxis of neurodegenerative disease, particularly in the treatment and prophylaxis of Parkinson's disease.

Claims

1. Cholestane derivatives of the general formula I ##STR00044## wherein, a is single or double bond; double bond is independently selected from in either E or Z configuration; R1, R2 is independently selected from the group consisting of hydrogen or hydroxyl group in alpha or beta configuration and their combination thereof; R3 is independently selected from the group consisting hydrogen, linear and branched C1-5 alkyl and cycloalkyl; R4 is independently selected from the group consisting hydrogen, methyl and ethyl; R3, R4 is independently selected from the bivalent group consisting C2-C5 α,ω-alkylene, and wherein the compounds 2α-hydroxy-5α-cholestan-6-one, 3α-hydroxy-5α-cholestan-6-one, 2α,3α-dihydroxy-5α-cholestan-6-one are excluded from the cholestane derivatives of general formula I, in the form of racemates and isomers and the pharmaceutically acceptable salts thereof, in particular salts with alkali metals, ammonium or amines, or addition salts with acids.

2. Cholestane derivatives according to claim 1 with linear C1-5 alkyl in position R3, which is selected from the group consisting of methyl, ethyl, n-propyl, n-butyl, and n-pentyl.

3. Cholestane derivatives according to claim 1 with branched C1-5 alkyl in position R3, which is selected from the group consisting of isopropyl, isobutyl, sec-butyl, tert-butyl, 2-methylbutan-2-yl, 2,2-dimethylpropyl, 3-methylbutyl, pentan-2-yl, pentan-3-yl, 3-methylbutan-2-yl, and 2-methylbutyl.

4. Cholestane derivatives according to claim 1 with cycloalkyl in position R3, which is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, which can be independently at each occurrence substituted by linear C1-5 alkyl.

5. Cholestane derivatives according to claim 1 with an alkylene group in position R3 and R, which are selected from the group consisting of 1,2-ethylene, trimethylene, tetramethylene, and pentamethylene.

6. Cholestane derivatives according to claim 1 with a chiral centre in position R3, which is selected from the group consisting of either R or S absolute configuration.

7. Cholestane derivatives according to claim 1, selected from the group consisting of 2α-hydroxy-24-nor-5α-cholan-6-on, 3α-hydroxy-24-nor-5α-cholan-6-one, 2α,3α-dihydroxy-24-nor-5α-cholan-6-one, 2α-hydroxy-5α-cholan-6-one, 3α-hydroxy-5α-cholan-6-one, 2α,3α-dihydroxy-5α-cholan-6-one, 2α-hydroxy-23-methyl-5α-cholan-6-one, 3α-hydroxy-23-methyl-5α-cholan-6-one, 2α,3α-dihydroxy-23-methyl-5α-cholan-6-one, 2α-hydroxy-26,27-dinor-5α-cholestan-6-one, 3α-hydroxy-26,27-dinor-5α-cholestan-6-one, 2α,3α-dihydroxy-26,27-dinor-5α-cholestan-6-one, (23R)-2α-hydroxy-23-ethyl-5α-cholan-6-one, (23R)-3α-hydroxy-23-ethyl-5α-cholan-6-one, (23R)-2α,3α-dihydroxy-23-ethyl-5α-cholan-6-one, (23S)-2α-hydroxy-23-ethyl-5α-cholan-6-one, (23S)-3α-hydroxy-23-ethyl-5α-cholan-6-one, (23S)-2α,3α-dihydroxy-23-ethyl-5α-cholan-6-one, 2α-hydroxy-27-nor-5α-cholestan-6-one, 3α-hydroxy-27-nor-5α-cholestan-6-one, 2α,3α-dihydroxy-27-nor-5α-cholestan-6-one, 2α-hydroxy-24,24-dimethyl-5α-cholan-6-one, 3α-hydroxy-24,24-dimethyl-5α-cholan-6-one, 2α,3α-dihydroxy-24,24-dimethyl-5α-cholan-6-one, 2α-hydroxy-26α-homo-27-nor-5α-cholestan-6-one, 3α-hydroxy-26α-homo-27-nor-5α-cholestan-6-one, 2α,3α-dihydroxy-26α-homo-27-nor-5α-cholestan-6-one, 2α-hydroxy-24,24,24-trimethyl-5α-cholan-6-one, 3α-hydroxy-24,24,24-trimethyl-5α-cholan-6-one, 2α,3α-dihydroxy-24,24,24-trimethyl-5α-cholan-6-one, 2α-hydroxy-27-nor-5α-campestan-6-one, 3α-hydroxy-27-nor-5α-campestan-6-one, 2α,3α-dihydroxy-27-nor-5α-campestan-6-one, 2α-hydroxy-27-nor-5α-ergostan-6-one, 3α-hydroxy-27-nor-5α-ergostan-6-one, 2α,3α-dihydroxy-27-nor-5α-ergostan-6-one; 2α-hydroxy-26α,26b-dihomo-27-nor-5α-cholestan-6-one, 3α-hydroxy-26α,26b-dihomo-27-nor-5α-cholestan-6-one, 2α,3α-dihydroxy-26α,26b-dihomo-27-nor-5α-cholestan-6-one, 2α-hydroxy-5α-campestan-6-one, 2α-hydroxy-5α-ergostan-6-one, 3α-hydroxy-5α-ergostan-6-one, 2α,3α-dihydroxy-5α-ergostan-6-one, 2α-hydroxy-24-ethyl-24,24-dimethyl-5α-cholan-6-one, 3α-hydroxy-24-ethyl-24,24-dimethyl-5α-cholan-6-one, 2α,3α-dihydroxy-24-ethyl-24,24-dimethyl-5α-cholan-6-one, 2α-hydroxy-25-methyl-5α-cholestan-6-one,3α-hydroxy-25-methyl-5α-cholestan-6-one, 2α,3α-dihydroxy-25-methyl-5α-cholestan-6-one, 2α-hydroxy-27-nor-5α-stigmastan-6-one, 3α-hydroxy-27-nor-5α-stigmastan-6-one, 2α,3α-dihydroxy-27-nor-5α-stigmastan-6-one, (24S)-2α-hydroxy-26α-homo-27-nor-5α-campestan-6-one, (24S)-3α-hydroxy-26α-homo-27-nor-5α-campestan-6-one, (24S)-2α,3α-dihydroxy-26α-homo-27-nor-5α-campestan-6-one, (24R)-2α-hydroxy-26α-homo-27-nor-5α-ergostan-6-one, (24R)-3α-hydroxy-26α-homo-27-nor-5α-ergostan-6-one, (24R)-2α,3α-dihydroxy-26α-homo-27-nor-5α-ergostan-6-one, 2α-hydroxy-22-cyclopropyl-23,24-dinor-5α-cholan-6-one, 3α-hydroxy-22-cyclopropyl-23,24-dinor-5α-cholan-6-one, 2α,3α-dihydroxy-22-cyclopropyl-23,24-dinor-5α-cholan-6-one, 2α-hydroxy-22-cyclobutyl-23,24-dinor-5α-cholan-6-one, 3α-hydroxy-22-cyclobutyl-23,24-dinor-5α-cholan-6-one, 2α,3α-dihydroxy-22-cyclobutyl-23,24-dinor-5α-cholan-6-one, 2α-hydroxy-22-cyclopentyl-23,24-dinor-5α-cholan-6-one, 3α-hydroxy-22-cyclopentyl-23,24-dinor-5α-cholan-6-one, 2α,3α-dihydroxy-22-cyclopentyl-23,24-dinor-5α-cholan-6-one, 2α-hydroxy-22-cyclohexyl-23,24-dinor-5α-cholan-6-one, 3α-hydroxy-22-cyclohexyl-23,24-dinor-5α-cholan-6-one, 2α,3α-dihydroxy-22-cyclohexyl-23,24-dinor-5α-cholan-6-one, 2α-hydroxy-23-cyclopropyl-24-nor-5α-cholan-6-one, 3α-hydroxy-23-cyclopropyl-24-nor-5α-cholan-6-one, 2α,3α-dihydroxy-23-cyclopropyl-24-nor-5α-cholan-6-one, 2α-hydroxy-23-cyclobutyl-24-nor-5α-cholan-6-one, 3α-hydroxy-23-cyclobutyl-24-nor-5α-cholan-6-one, 2α,3α-dihydroxy-23-cyclobutyl-24-nor-5α-cholan-6-one, 2α-hydroxy-23-cyclopentyl-24-nor-5α-cholan-6-one, 3α-hydroxy-23-cyclopentyl-24-nor-5α-cholan-6-one, 2α,3α-dihydroxy-23-cyclopentyl-24-nor-5α-cholan-6-one, 2α-hydroxy-23-cyclohexyl-24-nor-5α-cholan-6-one, 3α-hydroxy-23-cyclohexyl-24-nor-5α-cholan-6-one, 2α,3α-dihydroxy-23-cyclohexyl-24-nor-5α-cholan-6-one.

8. A method comprising administering a medicament comprising the cholestane derivatives of the general formula I according to claim 1.

9. A method of treatment comprising the step of administering the cholestane derivatives according to claim 1 for protection or prophylaxis of cell damage, injury and cell death.

10. A method of treatment comprising the step of administering the cholestane derivatives according to claim 1 for prophylaxis of neurodegenerative diseases.

11. The method of treatment according to claim 9, wherein the neurodegenerative disease is selected from amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, progressive supranuclear palsy, corticobasal degeneration, frontotemporal dementia, Lewy body dementia, multiple system atrophy, chronic traumatic encephalopathy, spinocerebellar ataxia.

12. A pharmaceutical composition comprising one or more cholestane derivatives according to claim 1 and at least one pharmaceutically acceptable excipient.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0072] FIG. 1 shows neuroprotective effect of novel compounds in glutamate-induced model of oxidative damage on dopaminergic neuron-like SH-SY5Y cells. The compounds of the invention at 0.1, 1 and 10 μM along with positive controls: R-lipoic acid (R-LA, 0.5, 5 and 50 μM) and deferoxamine (DFO, 1, and 100 μM) were used in co-treatment with 160 mM glutamate (Glu) for 24 hours. After 24 hours cell death was quantified by propidium iodide staining. Cell death generated by Glu was considered as 100% so the reduction of cell death was observed. All results are presented as mean±the standard error of the mean (SEM) in triplicate experiments (n=3) in three separated days. ANOVA, Tukey post hoc test; *, #P<0.05; **, ##P<0.01; ***, ###P<0.001; *P compared with vehicle with Glu 160 mM, #P compared with vehicle without Glu 160 mM. A value of P<0.05 is considered statistically significant.

[0073] FIG. 2 Glutamate-induced oxidative stress (OS) and OS-reducing activity of compounds of the invention and positive controls. The compounds of the invention at 0.1, 1 and 10 μM along with positive controls: R-lipoic acid (R-LA, 0.5, 5 and 50 μM) and deferoxamine (DFO, 1, 10 and 100 μM) were used in co-treatment with 160 mM Glu for 4 hours. After 4 hours superoxide radical formation as marker of oxidative stress was quantified by dihydroethidium staining. Oxidative stress generated by Glu was considered as 100% so the reduction of cell death was observed. All results are presented as mean±the standard error of the mean (SEM) in triplicate experiments (n=3) in three separated days. ANOVA, Tukey post hoc test; *, #P<0.05; **, ##P<0.01; ***, ###P<0.001; *P compared with vehicle with Glu 160 mM, #P compared with vehicle without Glu 160 mM. A value of P<0.05 is considered statistically significant.

EXAMPLES OF CARRYING OUT THE INVENTION

[0074] The following examples serve to illustrate the invention without limiting the scope thereof. Unless otherwise stated, all percentages and the like amounts are based on weight. The starting materials may be obtained from commercial sources (Sigma, Aldrich, Fluka, etc.) or can be prepared as described below.

[0075] 1H and 13C experiments were performed on Jeol ECX-500SS (500 MHz for 1H), and Varian.sup.UNITY Inova 400 (400 MHz for 1H) instruments, using CDCl.sub.3, DMSO-d.sub.6, CD.sub.3OD or THF-ds as solvents (25° C.). Chemical shifts (δ) were referenced to the residual signal of the solvent (CDCl.sub.3, DMSO-d.sub.6, CD.sub.3OD or THF-d.sub.8) and are reported in parts per million (ppm). Coupling constants (J) are reported in Hertz (Hz). NMR spectra were processed in the ACD/NMR Processor Academic Edition 12.01, MestReNova 6.0.2-5475 or JEOL Delta v5.0.5.1. HRMS analysis was performed using an LC-MS Orbitrap Elite high-resolution mass spectrometer with electrospray ionization (Dionex Ultimate 3000, Thermo Exactive plus, MA, USA). Spectra were taken at the positive and negative mode in the range of 100-1000 m/z. The samples were dissolved in MeOH and injected to the mass spectrometer over autosampler after HPLC separation: precolumn Phenomenex Gemini (C18, 50×2 mm, 2.6 μm), mobile phase isocratic MeOH/water/HCOOH 95:5:0.1. The course of the reactions was monitored by TLC on Kieselgel 60 F.sub.254 plates (Merck) detected by spraying with 10% aqueous H.sub.2SO.sub.4 and heating to 400° C. Purification was performed using column chromatography on Silica gel 60 (Merck 7734).

[0076] Starting steroid derivative 1 for olefination reaction was prepared from commercially available stigmasterol (lit. Heterocycles 1982, 17, 301). All new compounds described in this invention were prepared according to the following general scheme:

##STR00031##

[0077] Synthesis of New Cholane and Cholestane Derivatives

Example 1

[0078] ##STR00032##

2α,3α-(Isopropylidenedioxy)-6,6-(ethylenedioxy)-24-nor-5α-chol-22-en (compound 2a)

[0079] To a solution of methyltriphenylphosphonium bromide (48 mg; 0.134 mmol) in anhydrous tetrahydrofuran (3 mL) was added solution of n-butyllithium in hexane (1.6 M; 85 μL; 0.134 mmol). The mixture was stirred at 0° C. for 30 minutes. Then, the solution of aldehyde 1 (30 mg; 0.067 mmol) in anhydrous tetrahydrofuran (3 mL) was added and the reaction mixture was left to reach room temperature and stirred for additional 2 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (7% ethyl acetate in cyclohexane) to afford the title compound 2a.

[0080] Colourless oil, chemical formula: C.sub.28H.sub.44O.sub.4, yield: 25 mg, 84%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.69 (s, 3H, CH.sub.3); 0.84 (s, 3H, CH.sub.3); 1.02 (d, 3H, J=6.4 Hz, CH.sub.3); 1.32 (s, 3H, CH.sub.3); 1.47 (s, 3H, CH.sub.3); 1.92 (dd, 1H, J=12.7 and 6.9 Hz); 1.97 (dt, 1H, J=12.7 and 3.2 Hz); 2.06 (m, 1H); 2.16 (m, 1H); 3.75 (m, 1H, CH.sub.2O); 3.88-3.97 (m, 3H, CH.sub.2O); 4.10 (m, 1H, H-2(3); 4.27 (m, 1H, H-3(3); 4.81 (dd, 1H, J=10.2 and 1.9 Hz, H-23a); 4.90 (dd, 1H, J=17.1 and 1.9 Hz, H-23b); 5.65 (ddd, 1H, J=17.1, 10.2 and 8.7 Hz, H-22). .sup.13C NMR δ 12.14, 13.35, 20.04, 20.70, 21.94, 24.14, 26.55, 28.30, 28.59, 32.84, 37.98, 39.51, 41.01, 41.19, 42.43, 42.68, 45.47, 52.95, 55.37, 55.90, 64.15, 65.46, 72.82, 72.93, 107.51, 109.65, 111.54, 145.16. HRMS (API): m/z calcd for C.sub.28H.sub.45O.sub.41[M+H].sup.+ 445.3318, found 445.3319.

##STR00033##

2α,3α-Dihydroxy-24-nor-5α-chol-22-en-6-one (compound 3a)

[0081] 1M solution of hydrochloric acid (0.5 mL) was added to a solution of compound 2a (20 mg; 0.045 mmol) in tetrahydrofuran (4 mL) and the reaction mixture was heated at 40° C. for 4 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 3a.

[0082] White powder, chemical formula: C.sub.23H.sub.36O.sub.3, yield: 15 mg, 93%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.69 (s, 3H, CH.sub.3); 0.76 (s, 3H, CH.sub.3); 1.04 (d, 3H, J=6.7 Hz, CH.sub.3); 1.91 (dd, 1H, J=15.0 and 3.1 Hz); 1.97-2.11 (m, 2H); 2.29 (dd, 1H, J=13.3 and 4.4 Hz); 2.68 (dd, 1H, J=12.5 and 2.8 Hz); 3.77 (m, 1H, H-2β); 4.05 (m, 1H, H-3β); 4.83 (dd, 1H, J=10.1 and 1.9 Hz, H-23a); 4.91 (dd, 1H, J=17.1 and 1.1 Hz, H-23b); 5.65 (ddd, 1H, J=17.1, 10.1 and 8.6 Hz, H-22). .sup.13C NMR 612.17, 13.54, 20.04, 21.14, 23.90, 26.26, 28.15, 37.62, 39.24, 40.13, 41.11, 42.57, 42.88, 46.71, 50.67, 53.68, 55.25, 56.64, 68.24, 68.35, 111.84, 144.85, 212.23. HRMS (API): m/z calcd for C.sub.23H.sub.37O.sub.31[M+H].sup.+ 361.2743, found 361.2741.

##STR00034##

2α,3α-Dihydroxy-24-nor-5α-cholan-6-one (compound 4a)

[0083] Palladium on charcoal (12 mg) was added to a solution of compound 3a (12 mg; 0.033 mmol) in tetrahydrofuran (3 mL) and ethanol (1 mL). The flask with reaction mixture was evacuated and hydrogen was added from balloon. The reaction was stirred at room temperature for 18 hours. The palladium on charcoal was filtrated off, solvents were evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 4a.

[0084] White powder, chemical formula: C.sub.23H.sub.38O.sub.3, yield: 11 mg, 91%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.66 (s, 3H, CH.sub.3); 0.75 (s, 3H, CH.sub.3); 0.83 (t, 3H, J=7.2 Hz, CH.sub.3); 0.91 (d, 3H, J=6.4 Hz, CH.sub.3); 1.91 (dd, 1H, J=15.2 and 3.0 Hz); 1.96-2.08 (m, 2H); 2.30 (dd, 1H, J=13.3 and 4.5 Hz); 2.68 (dd, 1H, J=12.5 and 2.7 Hz); 3.76 (m, 1H, H-2β); 4.05 (m, 1H, H-3β). .sup.13C NMR δ 10.29, 11.98, 13.53, 17.98, 21.16, 23.92, 26.26, 28.17, 30.27, 36.88, 37.67, 39.35, 40.14, 42.58, 42.86, 46.77, 50.69, 53.69, 55.49, 56.62, 68.27, 68.37, 212.34. HRMS (API): m/z calcd for C.sub.23H.sub.39O.sub.3 [M+H].sup.+ 363.2899, found 363.2902.

Example 2

[0085] ##STR00035##

(22Z)-2α,3α-(Isopropylidenedioxy)-6,6-(ethylenedioxy)-5α-chol-22-en (compound 2b)

[0086] To a solution of ethyltriphenylphosphonium bromide (50 mg; 0.134 mmol) in anhydrous tetrahydrofuran (3 mL) was added solution of n-butyllithium in hexane (1.6 M; 85 μL; 0.134 mmol). The mixture was stirred at 0° C. for 30 minutes. Then, the solution of aldehyde 1 (30 mg; 0.067 mmol) in anhydrous tetrahydrofuran (3 mL) was added and the reaction mixture was left to reach room temperature and stirred for additional 2 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (7% ethyl acetate in cyclohexane) to afford the title compound 2b as predominantly Z isomer (less than 5% of E isomer (2bE) observed in .sup.1H NMR).

[0087] Colourless oil, chemical formula: C.sub.29H.sub.46O.sub.4, yield: 27 mg, 87%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.71 (s, 3H, CH.sub.3); 0.839 (s, 3H, CH.sub.3); 0.95 (d, 3H, J=6.7 Hz, CH.sub.3); 1.32 (s, 3H, CH.sub.3); 1.47 (s, 3H, CH.sub.3); 1.60 (dd, 3H, J=6.7 and 1.5 Hz, CH.sub.3); 1.93 (dd, 1H, J=12.7 and 6.9 Hz); 1.97 (m, 1H); 2.15 (m, 1H); 2.46 (m, 1H); 3.75 (m, 1H, CH.sub.2O); 3.88-3.97 (m, 3H, CH.sub.2O); 4.10 (m, 1H, H-2β); 4.27 (m, 1H, H-3β); 5.15 (m, 1H, H-22); 5.25 (m, 1H, H-23). .sup.13C NMR δ 12.25, 13.10, 13.37, 20.42, 20.71, 21.95, 24.13, 26.55, 27.88, 28.60, 32.85, 33.70, 38.00, 39.55, 41.03, 42.37, 42.70, 45.48, 53.01, 55.95, 56.16, 64.15, 65.47, 72.84, 72.95, 107.51, 109.67, 120.29, 137.50. HRMS (API): m/z calcd for C.sub.29H.sub.47O.sub.4 [M+H].sup.+ 459.3474, found 459.3475. Selected .sup.1H NMR signals for 22E-isomer (2bE): δ 0.67 (s, 3H, CH.sub.3); 0.833 (s, 3H, CH.sub.3); 0.98 (d, 3H, J=6.7 Hz, CH.sub.3); 5.29 (m, 1H).

##STR00036##

(22Z)-2α,3α-Dihydroxy-5α-chol-22-en-6-one (compound 3b)

[0088] 1M solution of hydrochloric acid (0.5 mL) was added to a solution of compound 2b (20 mg; 0.045 mmol) in tetrahydrofuran (4 mL) and the reaction mixture was heated at 40° C. for 4 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 3b (due to the presence of minor E-isomer (2bE) in the starting material 3bE (5%) can be observed in .sup.1H NMR).

[0089] White powder, chemical formula: C.sub.24H.sub.38O.sub.3, yield: 15 mg, 92%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.71 (s, 3H, CH.sub.3); 0.764 (s, 3H, CH.sub.3); 0.97 (d, 3H, J=6.7 Hz, CH.sub.3); 1.60 (dd, 3H, J=6.7 and 1.5 Hz, CH.sub.3); 1.92 (dd, 1H, J=15.1 and 3.2 Hz); 1.97-2.06 (m, 2H); 2.30 (dd, 1H, J=13.1 and 4.5 Hz); 2.47 (m, 1H); 2.69 (dd, 1H, J=12.5 and 2.8 Hz); 3.78 (m, 1H, H-2β); 4.05 (m, 1H, H-3β); 5.15 (m, 1H, H-22); 5.28 (m, 1H, H-23). .sup.13C NMR δ12.28, 13.13, 13.55, 20.40, 21.16, 23.89, 26.26, 27.75, 33.65, 37.65, 39.27, 40.17, 42.60, 42.83, 46.74, 50.68, 53.74, 56.04, 56.68, 68.26, 68.36, 120.58, 137.20, 212.22. HRMS (API): m/z calcd for C.sub.24H.sub.39O.sub.31M[M+H].sup.+ 375.2899, found 375.2897. Selected .sup.1H NMR signals for 22E-isomer (3bE): δ 0.67 (s, 3H, CH.sub.3); 0.757 (s, 3H, CH.sub.3); 1.00 (d, 3H, J=6.7 Hz, CH.sub.3); 5.34 (m, 1H).

##STR00037##

2α,3α-Dihydroxy-5α-cholan-6-one (compound 4b)

[0090] Palladium on charcoal (12 mg) was added to a solution of compound 3b with minor 3bE (12 mg; 0.033 mmol) in tetrahydrofuran (3 mL) and ethanol (1 mL). The flask with reaction mixture was evacuated and hydrogen was added from balloon. The reaction was stirred at room temperature for 18 hours. The palladium on charcoal was filtrated off, solvents were evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 4b.

[0091] White powder, chemical formula: C.sub.24H.sub.40O.sub.3, yield: 10 mg, 83%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.66 (s, 3H, CH.sub.3); 0.76 (s, 3H, CH.sub.3); 0.87 (t, 3H, J=7.0 Hz, CH.sub.3); 0.91 (d, 3H, J=6.4 Hz, CH.sub.3); 1.92 (dd, 1H, J=15.2 and 3.3 Hz); 1.97-2.08 (m, 2H); 2.30 (dd, 1H, J=13.2 and 4.6 Hz); 2.68 (dd, 1H, J=12.6 and 2.9 Hz); 3.77 (m, 1H, H-213); 4.05 (m, 1H, H-313). .sup.13C NMR δ 11.98, 13.54, 14.52, 18.56, 19.19, 21.18, 23.93, 26.26, 28.01, 35.49, 37.68, 38.20, 39.38, 40.17, 42.60, 42.92, 46.78, 50.69, 53.70, 56.05, 56.66, 68.29, 68.38, 212.26. HRMS (API): m/z calcd for C.sub.24H.sub.39O.sub.2 [M−H.sub.2O+H].sup.+ 359.2950, found 359.2948.

Example 3

[0092] ##STR00038##

(22Z)-2α,3α-(Isopropylidenedioxy)-6,6-(ethylenedioxy)-26,27-dinor-5α-cholest-22-en (compound 2c)

[0093] To a solution of propyltriphenylphosphonium bromide (52 mg; 0.135 mmol) in anhydrous tetrahydrofuran (3 mL) was added solution of n-butyllithium in hexane (1.6 M; 85 μL; 0.134 mmol). The mixture was stirred at 0° C. for 30 minutes. Then, the solution of aldehyde 1 (30 mg; 0.067 mmol) in anhydrous tetrahydrofuran (3 mL) was added and the reaction mixture was left to reach room temperature and stirred for additional 2 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (7% ethyl acetate in cyclohexane) to afford the title compound 2c as predominantly Z isomer (7% of E isomer (2cE) observed in .sup.1H NMR).

[0094] Colourless oil, chemical formula: C.sub.30H.sub.48O.sub.4, yield: 28 mg, 88%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.70 (s, 3H, CH.sub.3); 0.841 (s, 3H, CH.sub.3); 0.95 (t, 3H, J=7.4 Hz, CH.sub.3); 0.96 (d, 3H, J=6.7 Hz, CH.sub.3); 1.33 (s, 3H, CH.sub.3); 1.48 (s, 3H, CH.sub.3); 1.91-1.99 (m, 2H); 2.00-2.09 (m, 2H); 2.16 (m, 1H); 2.42 (m, 1H); 3.75 (m, 1H, CH.sub.2O); 3.88-3.97 (m, 3H, CH.sub.2O); 4.11 (m, 1H, H-2β); 4.27 (m, 1H, H-3β); 5.09 (m, 1H, H-22); 5.18 (m, 1H, H-23). .sup.13C NMR δ12.26, 13.38, 14.49, 20.71, 20.80, 20.86, 21.96, 24.13, 26.56, 28.06, 28.61, 32.86, 34.15, 38.00, 39.56, 41.03, 42.37, 42.70, 45.48, 53.01, 55.97, 56.02, 64.16, 65.48, 72.84, 72.95, 107.52, 109.68, 128.22, 135.93. HRMS (API): m/z calcd for C.sub.30H.sub.49O.sub.4 [M+H].sup.+ 473.3631, found 473.3633. Selected .sup.1H NMR signals for 22E-isomer (2cE): δ 0.67 (s, 3H, CH.sub.3); 0.838 (s, 3H, CH.sub.3); 0.99 (d, 3H, J=6.7 Hz, CH.sub.3); 5.33 (m, 1H).

##STR00039##

(22Z)-2α,3α-Dihydroxy-26,27-dinor-5α-cholest-22-en-6-one (compound 3c)

[0095] 1M solution of hydrochloric acid (0.5 mL) was added to a solution of compound 2c (20 mg; 0.042 mmol) in tetrahydrofuran (4 mL) and the reaction mixture was heated at 40° C. for 4 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 3c (due to the presence of minor E-isomer (2cE) in the starting material 3cE (7%) can be observed in .sup.1H NMR).

[0096] White powder, chemical formula: C25114.003, yield: 15 mg, 91%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.70 (s, 3H, CH.sub.3); 0.76 (s, 3H, CH.sub.3); 0.95 (t, 3H, J=7.4 Hz, CH.sub.3); 0.97 (d, 3H, J=6.7 Hz, CH.sub.3); 1.91 (dd, 1H, J=15.3 and 3.1 Hz); 1.97-2.07 (m, 2H); 2.29 (dd, 1H, J=13.1 and 4.6 Hz); 2.43 (m, 1H); 2.68 (dd, 1H, J=12.5 and 2.8 Hz); 3.77 (m, 1H, H-2β); 4.05 (m, 1H, H-3β); 5.09 (m, 1H, H-22); 5.19 (m, 1H, H-23). .sup.13C NMR δ 12.40, 13.68, 14.60, 20.89, 21.00, 21.28, 24.02, 26.40, 28.07, 34.22, 37.78, 39.41, 40.28, 42.73, 42.94, 46.87, 50.80, 53.85, 56.02, 56.82, 68.37, 68.47, 128.61, 135.74, 212.39. HRMS (API): m/z calcd for C.sub.25H.sub.41O.sub.3 [M+H].sup.+ 389.3056, found 389.3053. Selected .sup.1H NMR signals for 22E-isomer (3bE): δ 0.67 (s, 3H, CH.sub.3); 0.74 (s, 3H, CH.sub.3); 1.00 (d, 3H, J=6.7 Hz, CH.sub.3); 5.35 (m, 1H).

##STR00040##

2α,3α-Dihydroxy-26,27-dinor-5α-cholestan-6-one (compound 4c)

[0097] Palladium on charcoal (12 mg) was added to a solution of compound 3c with minor 3cE (12 mg; 0.031 mmol) in tetrahydrofuran (3 mL) and ethanol (1 mL). The flask with reaction mixture was evacuated and hydrogen was added from balloon. The reaction was stirred at room temperature for 18 hours. The palladium on charcoal was filtrated off, solvents were evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 4c.

[0098] White powder, chemical formula: C.sub.25H.sub.42O.sub.3, yield: 10 mg, 83%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.66 (s, 3H, CH.sub.3); 0.75 (s, 3H, CH.sub.3); 0.89 (t, 3H, J=7.0 Hz, CH.sub.3); 0.91 (d, 3H, J=6.4 Hz, CH.sub.3); 1.91 (dd, 1H, J=15.2 and 3.3 Hz); 1.96-2.07 (m, 2H); 2.30 (dd, 1H, J=13.2 and 4.6 Hz); 2.68 (dd, 1H, J=12.6 and 3.0 Hz); 3.76 (m, 1H, H-2β); 4.05 (m, 1H, H-3β). .sup.13C NMR δ11.98, 13.53, 14.17, 18.61, 21.17, 23.11, 23.92, 26.26, 27.99, 28.28, 30.28, 35.50, 37.68, 39.37, 40.16, 42.59, 42.90, 46.78, 50.69, 53.70, 55.97, 56.65, 68.28, 68.38, 212.32. HRMS (API): m/z calcd for C.sub.25H.sub.43O.sub.2 [M−H.sub.2O+H].sup.+ 391.3212, found 391.3209.

Example 4

[0099] ##STR00041##

(22Z)-2α,3α-(Isopropylidenedioxy)-6,6-(ethylenedioxy)-27-nor-5α-cholest-22-en (compound 2d)

[0100] To a solution of butyltriphenylphosphonium bromide (54 mg; 0.135 mmol) in anhydrous tetrahydrofuran (3 mL) was added solution of n-butyllithium in hexane (1.6 M; 85 μL; 0.134 mmol). The mixture was stirred at 0° C. for 30 minutes. Then, the solution of aldehyde 1 (30 mg; 0.067 mmol) in anhydrous tetrahydrofuran (3 mL) was added and the reaction mixture was left to reach room temperature and stirred for additional 2 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (7% ethyl acetate in cyclohexane) to afford the title compound 2d as predominantly Z isomer (20% of E isomer (2dE) observed in .sup.1H NMR).

[0101] Colourless oil, chemical formula: C311-15004, yield: 27 mg, 82%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.70 (s, 3H, CH.sub.3); 0.84 (s, 3H, CH.sub.3); 0.89 (t, 3H, J=7.0 Hz, CH.sub.3); 0.91 (d, 3H, J=6.7 Hz, CH.sub.3); 1.33 (s, 3H, CH.sub.3); 1.48 (s, 3H, CH.sub.3); 1.89-2.09 (m, 5H); 2.15 (m, 1H); 2.42 (m, 1H); 3.75 (m, 1H, CH.sub.2O); 3.88-3.97 (m, 3H, CH.sub.2O); 4.10 (m, 1H, H-2β); 4.28 (m, 1H, H-3β); 5.11-5.20 (m, 2H H-22, H-23). .sup.13C NMR δ 12.28, 13.39, 13.91, 20.72, 20.74, 21.97, 22.98, 24.14, 26.57, 28.03, 28.61, 29.67, 32.86, 34.19, 38.01, 39.57, 41.03, 42.37, 42.71, 45.49, 53.02, 55.97, 56.10, 64.17, 65.48, 72.84, 72.96, 107.52, 109.68, 126.44, 136.64. HRMS (API): m/z calcd for C.sub.31H.sub.51O.sub.4 [M+H].sup.+ 487.3787, found 487.3783. Selected 1H NMR signals for 22E-isomer (2dE): δ 0.68 (s, 3H, CH.sub.3); 0.84 (s, 3H, CH.sub.3); 0.87 (t, 3H, J=7.0 Hz, CH.sub.3); 1.00 (d, 3H, J=6.7 Hz, CH.sub.3); 5.28 (m, 1H).

##STR00042##

(22Z)-2α,3α-Dihydroxy-27-nor-5α-cholest-22-en-6-one (compound 3d)

[0102] 1M solution of hydrochloric acid (0.5 mL) was added to a solution of compound 2d (20 mg; 0.041 mmol) in tetrahydrofuran (4 mL) and the reaction mixture was heated at 40° C. for 4 hours. The reaction mixture was diluted with ethyl acetate and extracted twice with water. Organic layer was dried over calcium chloride. Solvent was evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 3c (due to the presence of minor E-isomer (2dE) in the starting material 3dE (20%) can be observed in .sup.1H NMR). White powder, chemical formula: C25114.003, yield: 15 mg, 91%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.70 (s, 3H, CH.sub.3); 0.764 (s, 3H, CH.sub.3); 0.91 (t, 3H, J=7.4 Hz, CH.sub.3); 0.97 (d, 3H, J=6.7 Hz, CH.sub.3); 1.92 (m, 1H); 1.96-2.07 (m, 4H); 2.29 (dd, 1H, J=13.1 and 4.4 Hz); 2.44 (m, 1H); 2.69 (dd, 1H, J=12.6 and 2.7 Hz); 3.78 (m, 1H, H-2β); 4.05 (m, 1H, H-3β); 5.11-5.24 (m, 2H, H-22, H-23). .sup.13C NMR δ 12.29, 13.56, 13.91, 20.71, 21.16, 22.96, 23.90, 26.26, 27.91, 29.68, 34.14, 37.66, 39.28, 40.16, 42.60, 42.81, 46.74, 50.67, 53.74, 55.97, 56.69, 68.26, 68.35, 126.72, 136.33, 212.25. HRMS (API): m/z calcd for C.sub.26H.sub.43O.sub.3 [M+H].sup.+ 403.3212, found 403.3211. Selected .sup.1H NMR signals for 22E-isomer (3bE): δ 0.67 (s, 3H, CH.sub.3); 0.758 (s, 3H, CH.sub.3); 0.87 (t, 3H, J=7.0 Hz, CH.sub.3); 1.01 (d, 3H, J=6.7 Hz, CH.sub.3); 5.28 (m, 1H).

##STR00043##

2α,3α-Dihydroxy-27-nor-5α-cholestan-6-one (compound 4d)

[0103] Palladium on charcoal (12 mg) was added to a solution of compound 3d with minor 3dE (12 mg; 0.030 mmol) in tetrahydrofuran (3 mL) and ethanol (1 mL). The flask with reaction mixture was evacuated and hydrogen was added from balloon. The reaction was stirred at room temperature for 18 hours. The palladium on charcoal was filtrated off, solvents were evaporated under reduce pressure and crude product was subjected to silica gel chromatography (60% ethyl acetate in cyclohexane) to afford the title compound 4d.

[0104] White powder, chemical formula: C.sub.26H.sub.44O.sub.3, yield: 10 mg, 83%. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ, ppm: δ 0.66 (s, 3H, CH.sub.3); 0.76 (s, 3H, CH.sub.3); 0.89 (t, 3H, J=7.0 Hz, CH.sub.3); 0.91 (d, 3H, J=6.4 Hz, CH.sub.3); 1.92 (dd, 1H, J=15.2 and 3.3 Hz); 1.96-2.07 (m, 2H); 2.30 (dd, 1H, J=13.2 and 4.6 Hz); 2.68 (dd, 1H, J=12.6 and 3.0 Hz); 3.77 (m, 1H, H-2β); 4.05 (m, 1H, H-3β). .sup.13C NMR δ 11.98, 13.54, 14.13, 18.61, 21.17, 22.73, 23.93, 25.70, 26.26, 27.99, 30.28, 35.64, 35.77, 37.68, 39.37, 40.16, 42.59, 42.91, 46.78, 50.69, 53.70, 55.98, 56.65, 68.29, 68.38, 212.27. HRMS (API): m/z calcd for C.sub.26H.sub.43O.sub.2 [M−H.sub.2O+H].sup.+ 387.3263, found 387.3260.

TABLE-US-00001 TABLE 1 Examples of cholestane derivatives (Formula 1) substituents CH analysis MS analysis R1 R2 R3 R4 a [% C, % H] [calc./found] [M + H].sup.+ 1 H OH H H double 80.18/80.16, 10.53/10.54 345 2 OH H H H double 80.18/80.16, 10.53/10.55 345 3 OH OH H H double 76.62/76.60, 10.06/10.09 361 4 H OH H H single 79.71/79.68, 11.05/11.08 347 5 OH H H H single 79.71/79.69, 11.05/11.06 347 6 OH OH H H single 76.20/76.18, 10.56/10.59 363 7 H OH methyl H double 80.39/80.38, 10.68/10.70 359 8 OH H methyl H double 80.39/80.38, 10.68/10.72 359 9 OH OH methyl H double 76.96/76.94, 10.23/10.25 375 10 H OH H methyl double 80.39/80.40, 10.68/10.70 359 11 OH H H methyl double 80.39/80.39, 10.68/10.71 359 12 OH OH H methyl double 76.96/76.93, 10.23/10.27 375 13 H OH methyl H single 79.94/79.91, 11.18/11.21 361 14 OH H methyl H single 79.94/79.93, 11.18/11.20 361 15 OH OH methyl H single 76.55/76.51, 10.71/10.74 377 16 H OH methyl methyl double 80.59/80.56, 10.82/10.85 373 17 OH H methyl methyl double 80.59/80.58, 10.82/10.83 373 18 OH OH methyl methyl double 77.27/77.25, 10.38/10.40 389 19 H OH methyl methyl single 80.16/80.14, 11.30/11.33 375 20 OH H methyl methyl single 80.16/80.13, 11.30/11.33 375 21 OH OH methyl methyl single 76.87/76.85, 10.84/10.88 391 22 H OH ethyl H double 80.59/80.57, 10.82/10.85 373 23 OH H ethyl H double 80.59/80.56, 10.82/10.86 373 24 OH OH ethyl H double 77.27/77.25, 10.38/10.40 389 25 H OH H ethyl double 80.59/80.57, 10.82/10.84 373 26 OH H H ethyl double 80.59/80.58, 10.82/10.83 373 27 OH OH H ethyl double 77.27/77.26, 10.38/10.40 389 28 H OH ethyl H single 80.16/80.14, 11.30/11.33 375 29 OH H ethyl H single 80.16/80.15, 11.30/11.34 375 30 OH OH ethyl H single 76.87/76.85, 10.84/10.86 391 31 H OH ethyl ethyl double 80.94/80.91, 11.07/11.10 401 32 OH H ethyl ethyl double 80.94/80.92, 11.07/11.08 401 33 OH OH ethyl ethyl double 77.83/77.81, 10.64/10.66 417 34 H OH ethyl ethyl single 80.54/80.53, 11.51/11.52 403 35 OH H ethyl ethyl single 80.54/80.52, 11.51/11.51 403 36 OH OH ethyl ethyl single 77.46/77.44, 11.07/11.09 419 37 H OH methyl ethyl double 80.77/80.72, 10.95/11.01 387 38 OH H methyl ethyl double 80.77/80.73, 10.95/11.00 387 39 OH OH methyl ethyl double 77.56/77.52, 10.51/10.55 403 40 H OH ethyl methyl double 80.77/80.74, 10.95/10.99 387 41 OH H ethyl methyl double 80.77/80.73, 10.95/11.01 387 42 OH OH ethyl methyl double 77.56/77.51, 10.51/10.55 403 43 H OH ethyl (23R)-methyl single 80.35/80.31, 11.41/11.46 389 44 OH H ethyl (23R)-methyl single 80.35/80.30, 11.41/11.46 389 45 OH OH ethyl (23R)-methyl single 77.18/77.15, 10.96/10.99 405 46 H OH ethyl (23S)-methyl single 80.35/80.30, 11.41/11.47 389 47 OH H ethyl (23S)-methyl single 80.35/80.30, 11.41/11.47 389 48 OH OH ethyl (23S)-methyl single 77.18/77.14, 10.96/11.01 405 49 H OH n-propyl H double 80.77/80.76, 10.95/10.98 387 50 OH H n-propyl H double 80.77/80.76, 10.95/10.99 387 51 OH OH n-propyl H double 77.56/77.54, 10.51/10.53 403 52 H OH H n-propyl double 80.77/80.75, 10.95/10.97 387 53 OH H H n-propyl double 80.77/80.76, 10.95/10.98 387 54 OH OH H n-propyl double 77.56/77.53, 10.51/10.54 403 55 H OH n-propyl H single 80.35/80.31, 11.41/11.45 389 56 OH H n-propyl H single 80.35/80.32, 11.41/11.43 389 57 OH OH n-propyl H single 77.18/77.16, 10.96/11.00 405 58 H OH isopropyl H double 80.77/80.75, 10.95/10.98 387 59 OH H isopropyl H double 80.77/80.78, 10.95/10.99 387 60 OH OH isopropyl H double 77.56/77.53, 10.51/10.53 403 61 H OH H isopropyl double 80.77/80.76, 10.95/10.98 387 62 OH H H isopropyl double 80.77/80.74, 10.95/10.99 387 63 OH OH H isopropyl double 77.56/77.53, 10.51/10.55 403 64 H OH isopropyl H single 80.35/80.33, 11.41/11.43 389 65 OH H isopropyl H single 80.35/80.33, 11.41/11.45 389 66 OH OH isopropyl H single 77.18/77.15, 10.96/10.99 405 67 H OH n-butyl H double 80.94/80.92, 11.07/11.09 401 68 OH H n-butyl H double 80.94/80.93, 11.07/11.10 401 69 OH OH n-butyl H double 77.83/77.80, 10.64/10.67 417 70 H OH H n-butyl double 80.94/80.92, 11.07/11.08 401 71 OH H H n-butyl double 80.94/80.91, 11.07/11.11 401 72 OH OH H n-butyl double 77.83/77.81, 10.64/10.66 417 73 H OH n-butyl H single 80.54/80.52, 11.51/11.54 403 74 OH H n-butyl H single 80.54/80.53, 11.51/11.52 403 75 OH OH n-butyl H single 77.46/77.44, 11.07/11.09 419 76 H OH (R)-but-2-yl H double 80.94/80.92, 11.07/11.10 401 77 OH H (R)-but-2-yl H double 80.94/80.91, 11.07/11.11 401 78 OH OH (R)-but-2-yl H double 77.83/77.82, 10.64/10.65 417 79 H OH H (R)-but-2-yl double 80.94/80.91, 11.07/11.09 401 80 OH H H (R)-but-2-yl double 80.94/80.91, 11.07/11.10 401 81 OH OH H (R)-but-2-yl double 77.83/77.81, 10.64/10.65 417 82 H OH (R)-but-2-yl H single 80.54/80.53, 11.51/11.52 403 83 OH H (R)-but-2-yl H single 80.54/80.51, 11.51/11.55 403 84 OH OH (R)-but-2-yl H single 77.46/77.45, 11.07/11.08 419 85 H OH (S)-but-2-yl H double 80.94/80.92, 11.07/11.08 401 86 OH H (S)-but-2-yl H double 80.94/80.93, 11.07/11.09 401 87 OH OH (S)-but-2-yl H double 77.83/77.80, 10.64/10.68 417 88 H OH H (S)-but-2-yl double 80.94/80.93, 11.07/11.10 401 89 OH H H (S)-but-2-yl double 80.94/80.91, 11.07/11.11 401 90 OH OH H (S)-but-2-yl double 77.83/77.81, 10.64/10.67 417 91 H OH (S)-but-2-yl H single 80.54/80.52, 11.51/11.53 403 92 OH H (S)-but-2-yl H single 80.54/80.52, 11.51/11.54 403 93 OH OH (S)-but-2-yl H single 77.46/77.45, 11.07/11.09 419 94 H OH tert-butyl H double 80.94/80.93, 11.07/11.08 401 95 OH H tert-butyl H double 80.94/80.92, 11.07/11.09 401 96 OH OH tert-butyl H double 77.83/77.81, 10.64/10.66 417 97 H OH H tert-butyl double 80.94/80.91, 11.07/11.11 401 98 OH H H tert-butyl double 80.94/80.92, 11.07/11.10 401 99 OH OH H tert-butyl double 77.83/77.82, 10.64/10.65 417 100 H OH tert-butyl H single 80.54/80.52, 11.51/11.54 403 101 OH H tert-butyl H single 80.54/80.53, 11.51/11.54 403 102 OH OH tert-butyl H single 77.46/77.45, 11.07/11.08 419 103 H OH isobutyl H double 80.94/80.92, 11.07/11.11 401 104 OH H isobutyl H double 80.94/80.91, 11.07/11.11 401 105 OH OH isobutyl H double 77.83/77.82, 10.64/10.65 417 106 H OH H isobutyl double 80.94/80.93, 11.07/11.08 401 107 OH H H isobutyl double 80.94/80.91, 11.07/11.09 401 108 OH OH H isobutyl double 77.83/77.81, 10.64/10.67 417 109 H OH isobutyl H single 80.54/80.51, 11.51/11.55 403 110 OH H isobutyl H single 80.54/80.53, 11.51/11.53 403 111 OH OH isobutyl H single 77.46/77.44, 11.07/11.10 419 112 H OH n-pentyl H double 81.10/81.07, 11.18/11.20 415 113 OH H n-pentyl H double 81.10/81.08, 11.18/11.20 415 114 OH OH n-pentyl H double 78.09/78.06, 10.77/10.79 431 115 H OH H n-pentyl double 81.10/81.06, 11.18/11.21 415 116 OH H H n-pentyl double 81.10/81.07, 11.18/11.19 415 117 OH OH H n-pentyl double 78.09/78.06, 10.77/10.80 431 118 H OH n-pentyl H single 80.71/80.68, 11.61/11.65 417 119 OH H n-pentyl H single 80.71/80.67, 11.61/11.66 417 120 OH OH n-pentyl H single 77.72/77.70, 11.18/11.21 433 121 H OH 3-methylbut-1-yl H double 81.10/81.07, 11.18/11.20 415 122 OH H 3-methylbut-1-yl H double 81.10/81.08, 11.18/11.19 415 123 OH OH 3-methylbut-1-yl H double 78.09/78.08, 10.77/10.78 431 124 H OH H 3-methylbut-1-yl double 81.10/81.08, 11.18/11.19 415 125 OH H H 3-methylbut-1-yl double 81.10/81.08, 11.18/11.20 415 126 OH OH H 3-methylbut-1-yl double 78.09/78.08, 10.77/10.78 431 127 H OH 3-methylbut-1-yl H single 80.71/80.69, 11.61/11.64 417 128 OH H 3-methylbut-1-yl H single 80.71/80.68, 11.61/11.65 417 129 OH OH 3-methylbut-1-yl H single 77.72/77.70, 11.18/11.20 433 130 H OH (R)-2-methylbut-1-yl H double 81.10/81.08, 11.18/11.20 415 131 OH H (R)-2-methylbut-1-yl H double 81.10/81.09, 11.18/11.19 415 132 OH OH (R)-2-methylbut-1-yl H double 78.09/78.07, 10.77/10.79 431 133 H OH H (R)-2-methylbut-1-yl double 81.10/81.06, 11.18/11.22 415 134 OH H H (R)-2-methylbut-1-yl double 81.10/81.07, 11.18/11.21 415 135 OH OH H (R)-2-methylbut-1-yl double 78.09/78.08, 10.77/10.78 431 136 H OH (R)-2-methylbut-1-yl H single 80.71/80.68, 11.61/11.64 417 137 OH H (R)-2-methylbut-1-yl H single 80.71/80.69, 11.61/11.63 417 138 OH OH (R)-2-methylbut-1-yl H single 77.72/77.69, 11.18/11.21 433 139 H OH (S)-2-methylbut-1-yl H double 81.10/81.08, 11.18/11.19 415 140 OH H (S)-2-methylbut-1-yl H double 81.10/81.08, 11.18/11.20 415 141 OH OH (S)-2-methylbut-1-yl H double 78.09/78.07, 10.77/10.79 431 142 H OH H (S)-2-methylbut-1-yl double 81.10/81.07, 11.18/11.21 415 143 OH H H (S)-2-methylbut-1-yl double 81.10/81.09, 11.18/11.20 415 144 OH OH H (S)-2-methylbut-1-yl double 78.09/78.08, 10.77/10.78 431 145 H OH (S)-2-methylbut-1-yl H single 80.71/80.68, 11.61/11.64 417 146 OH H (S)-2-methylbut-1-yl H single 80.71/80.68, 11.61/11.63 417 147 OH OH (S)-2-methylbut-1-yl H single 77.72/77.69, 11.18/11.22 433 148 H OH (R)-3-methylbut-2-yl H double 81.10/81.08, 11.18/11.19 415 149 OH H (R)-3-methylbut-2-yl H double 81.10/81.09, 11.18/11.19 415 150 OH OH (R)-3-methylbut-2-yl H double 78.09/78.06, 10.77/10.79 431 151 H OH H (R)-3-methylbut-2-yl double 81.10/81.08, 11.18/11.19 415 152 OH H H (R)-3-methylbut-2-yl double 81.10/81.07, 11.18/11.21 415 153 OH OH H (R)-3-methylbut-2-yl double 78.09/78.07, 10.77/10.79 431 154 H OH (R)-3-methylbut-2-yl H single 80.71/80.67, 11.61/11.65 417 155 OH H (R)-3-methylbut-2-yl H single 80.71/80.68, 11.61/11.64 417 156 OH OH (R)-3-methylbut-2-yl H single 77.72/77.70, 11.18/11.20 433 157 H OH (S)-3-methylbut-2-yl H double 81.10/81.09, 11.18/11.19 415 158 OH H (S)-3-methylbut-2-yl H double 81.10/81.09, 11.18/11.20 415 159 OH OH (S)-3-methylbut-2-yl H double 78.09/78.05, 10.77/10.80 431 160 H OH H (S)-3-methylbut-2-yl double 81.10/81.08, 11.18/11.20 415 161 OH H H (S)-3-methylbut-2-yl double 81.10/81.09, 11.18/11.21 415 162 OH OH H (S)-3-methylbut-2-yl double 78.09/78.06, 10.77/10.80 431 163 H OH (S)-3-methylbut-2-yl H single 80.71/80.69, 11.61/11.64 417 164 OH H (S)-3-methylbut-2-yl H single 80.71/80.70, 11.61/11.63 417 165 OH OH (S)-3-methylbut-2-yl H single 77.72/77.69, 11.18/11.23 433 166 H OH (R)-pent-2-yl H double 81.10/81.07, 11.18/11.20 415 167 OH H (R)-pent-2-yl H double 81.10/81.08, 11.18/11.20 415 168 OH OH (R)-pent-2-yl H double 78.09/78.07, 10.77/10.78 431 169 H OH H (R)-pent-2-yl double 81.10/81.08, 11.18/11.20 415 170 OH H H (R)-pent-2-yl double 81.10/81.07, 11.18/11.21 415 171 OH OH H (R)-pent-2-yl double 78.09/78.05, 10.77/10.80 431 172 H OH (R)-pent-2-yl H single 80.71/80.69, 11.61/11.64 417 173 OH H (R)-pent-2-yl H single 80.71/80.68, 11.61/11.64 417 174 OH OH (R)-pent-2-yl H single 77.72/77.70, 11.18/11.22 433 175 H OH (S)-pent-2-yl H double 81.10/81.06, 11.18/11.22 415 176 OH H (S)-pent-2-yl H double 81.10/81.07, 11.18/11.21 415 177 OH OH (S)-pent-2-yl H double 78.09/78.06, 10.77/10.80 431 178 H OH H (S)-pent-2-yl double 81.10/81.07, 11.18/11.21 415 179 OH H H (S)-pent-2-yl double 81.10/81.08, 11.18/11.20 415 180 OH OH H (S)-pent-2-yl double 78.09/78.07, 10.77/10.78 431 181 H OH (S)-pent-2-yl H single 80.71/80.69, 11.61/11.65 417 182 OH H (S)-pent-2-yl H single 80.71/80.67, 11.61/11.66 417 183 OH OH (S)-pent-2-yl H single 77.72/77.68, 11.18/11.23 433 184 H OH pent-3-yl H double 81.10/81.06, 11.18/11.22 415 185 OH H pent-3-yl H double 81.10/81.07, 11.18/11.21 415 186 OH OH pent-3-yl H double 78.09/78.05, 10.77/10.79 431 187 H OH H pent-3-yl double 81.10/81.08, 11.18/11.19 415 188 OH H H pent-3-yl double 81.10/81.08, 11.18/11.20 415 189 OH OH H pent-3-yl double 78.09/78.07, 10.77/10.80 431 190 H OH pent-3-yl H single 80.71/80.67, 11.61/11.66 417 191 OH H pent-3-yl H single 80.71/80.67, 11.61/11.65 417 192 OH OH pent-3-yl H single 77.72/77.70, 11.18/11.22 433 193 H OH 1,2-ethylene double 81.03/81.05, 10.34/10.38 371 194 OH H 1,2-ethylene double 81.03/81.07, 10.34/10.35 371 195 OH OH 1,2-ethylene double 77.68/77.65, 9.91/9.96 387 196 H OH 1,2-ethylene single 80.59/80.55, 10.82/10.83 373 197 OH H 1,2-ethylene single 80.59/80.54, 10.82/10.79 373 198 OH OH 1,2-ethylene single 77.27/77.30, 10.38/10.40 389 199 H OH trimethylene double 81.20/81.18, 10.48/10.50 385 200 OH H trimethylene double 81.20/81.17, 10.48/10.52 385 201 OH OH trimethylene double 77.95/77.91, 10.06/10.10 401 202 H OH trimethylene single 80.77/80.74, 10.95/10.96 387 203 OH H trimethylene single 80.77/80.75, 10.95/10.98 387 204 OH OH trimethylene single 77.56/77.52, 10.51/10.53 403 205 H OH tetramethylene double 81.35/81.31, 10.62/10.63 399 206 OH H tetramethylene double 81.35/81.33, 10.62/10.63 399 207 OH OH tetramethylene double 78.21/78.18, 10.21/10.25 415 208 H OH tetramethylene single 80.94/80.92, 11.07/11.10 401 209 OH H tetramethylene single 80.94/80.93, 11.07/11.09 401 210 OH OH tetramethylene single 77.83/77.81, 10.64/10.66 417 211 H OH pentamethylene double 81.50/81.47, 10.75/10.77 413 212 OH H pentamethylene double 81.50/81.49, 10.75/10.76 413 213 OH OH pentamethylene double 78.46/78.44, 10.35/10.38 429 214 H OH pentamethylene single 81.10/81.08, 11.18/11.20 415 215 OH H pentamethylene single 81.10/81.06, 11.18/11.22 415 216 OH OH pentamethylene single 78.09/78.05, 10.77/10.79 431 217 H OH cyclopropyl H double 81.20/81.18, 10.48/10.52 385 218 OH H cyclopropyl H double 81.20/81.17, 10.48/10.52 385 219 OH OH cyclopropyl H double 77.95/77.90, 10.06/10.09 401 220 H OH H cyclopropyl double 81.20/81.17, 10.48/10.54 385 221 OH H H cyclopropyl double 81.20/81.18, 10.48/10.54 385 222 OH OH H cyclopropyl double 77.95/77.91, 10.06/10.09 401 223 H OH cyclopropyl H single 80.77/80.74, 10.95/11.00 387 224 OH H cyclopropyl H single 80.77/80.73, 10.95/11.02 387 225 OH OH cyclopropyl H single 77.56/77.51, 10.51/10.56 403 226 H OH cyclobutyl H double 81.35/81.29, 10.62/10.66 399 227 OH H cyclobutyl H double 81.35/81.31, 10.62/10.65 399 228 OH OH cyclobutyl H double 78.21/78.17, 10.21/10.26 415 229 H OH H cyclobutyl double 81.35/81.29, 10.62/10.67 399 230 OH H H cyclobutyl double 81.35/81.28, 10.62/10.67 399 231 OH OH H cyclobutyl double 78.21/78.18, 10.21/10.24 415 232 H OH cyclobutyl H single 80.94/80.92, 11.07/11.10 401 233 OH H cyclobutyl H single 80.94/80.90, 11.07/11.11 401 234 OH OH cyclobutyl H single 77.83/77.80, 10.64/10.67 417 235 H OH cyclopentyl H double 81.50/81.45, 10.75/10.79 413 236 OH H cyclopentyl H double 81.50/81.46, 10.75/10.79 413 237 OH OH cyclopentyl H double 78.46/78.41, 10.35/10.40 429 238 H OH H cyclopentyl double 81.50/81.44, 10.75/10.80 413 239 OH H H cyclopentyl double 81.50/81.44, 10.75/10.79 413 240 OH OH H cyclopentyl double 78.46/78.43, 10.35/10.38 429 24 H OH cyclopentyl H single 81.10/81.04, 11.18/11.23 415 242 OH H cyclopentyl H single 81.10/81.06, 11.18/11.21 415 243 OH OH cyclopentyl H single 78.09/78.06, 10.77/10.81 431 244 H OH cyclohexyl H double 81.63/81.59, 10.87/10.91 427 245 OH H cyclohexyl H double 81.63/81.58, 10.87/10.92 427 246 OH OH cyclohexyl H double 78.68/78.64, 10.47/10.50 443 247 H OH H cyclohexyl double 81.63/81.59, 10.87/10.92 427 248 OH H H cyclohexyl double 81.63/81.60, 10.87/10.90 427 249 OH OH H cyclohexyl double 78.68/78.63, 10.47/10.52 443 250 H OH cyclohexyl H single 81.25/81.20, 11.29/11.34 429 251 OH H cyclohexyl H single 81.25/81.19, 11.29/11.35 429 252 OH OH cyclohexyl H single 78.33/78.28, 10.88/10.92 445

Example 6—Viability Tests on Human Neuron-Like SH-SY5Y Cells

[0105] The SH-SY5Y human neuroblastoma cell line obtained purchased from ECACC (The European Collection of Authenticated Cell Cultures) was cultivated in Dulbecco's modified Eagle's Medium and Ham's F12 Nutrient Mixture (DMEM:F-12, 1:1), supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin at 37° C. in a humidified atmosphere 5% CO.sub.2, 95% air in passage limit up to ECACC+20. The assay was performed in 96-well microplate with 7000 SH-SY5Y cells per well. Next day, SH-SY5Y underwent all-trans retinoic acid (ATRA)-differentiation for 48 h (ATRA 10 μM). After 48 h, old DMEM/F12 media was removed by fresh media containing tested compounds at 0.1, 1 and 10 μM concentrations for 24 h. The tested compounds were dissolved in DMSO were added into the medium. The maximum concentration of DMSO in media was kept below 0.1% (v/v). The cell viability was measured by Calcein AM (1 mg/ml ThermoFisher) viability assay. Solution of Calcein AM in PBS (0.75 μM) was pipetted to cells and incubated for 50 minutes. After that the fluorescence was measured at 488/517 nm (excitation/emission) using microplate reader Infinite M200 (TECAN). Calcein AM assay is based on the dye-intracellular-esterase cleavage of non-fluorescent dye (Calcein AM) by living cells to fluorescent dye (Calcein), while dyeing cells that lose such ability. The values in Table 4 show % of viability, relative to control of all tested compounds. The control (medium with DMSO, <0.1% (v/v)) was postulated as 100% viability (see the first line of the table). As shown in table 4, all derivatives were prove to not induce a decrease in viability.

TABLE-US-00002 TABLE 4 The effect of new derivatives on viability of human neuroblastoma cell line SH-SY5Y (neuron-like phenotype) after 24 hours of treatment. The data are expressed as percentages of DMSO control as the mean ± SEM of two independent experiments. Viability (% of DMSO control) DMSO control 100 0.95 Compound 0.1 μM SEM± 1 μM SEM± 10 μM SEM± 57 105.90 2.86 108.90 3.93 117.10 3.31 30 99.41 3.13 102.30 4.14 104.70 3.13 15 98.20 2.53 106.40 1.82 120.60 2.26 111 106.90 4.19 107.80 3.02 109.90 1.37 6 101.30 3.94 104.80 4.84 122.10 4.86 0.5 μM SEM± 5 μM SEM± 50 μM SEM± R-LA 96.12 2.68 96.93 3.59 101.2 2.09 1 μM SEM± 5 μM SEM± 50 μM SEM± DFO 105.7 8.48 105.5 8.49 109.5 5.92 *DMSO—dimethylsuplhoxide

Example 17 Neuroprotective Effect of Examples on Glutamate-Induced Oxidative Damage of Human Neuron-Like SH-SYSY Cells

[0106] Similarly, to the previous test the assay was performed in 96-well microplates with 20000 cells per well. Day after the seeding of cells, SH-SYSY underwent all-trans retinoic acid (ATRA)-differentiation for 48 h (ATRA 10 μM). After 48 h, old DMEM/F12 media was removed by fresh media containing 160 mM glutamate (Glu) without or with tested compounds at 0.1, 1 and 10 μM concentrations. As positive controls R-lipoic acid (R-LA) at 0.5, 5, 50 μM and deferoxamine (DFO) at 1, 10 and 100 μM were used. After 24 h the cell death was quantified by propidium iodide staining according to literature (Stone et al. BMC Cell Biol. 2003, 4, 1) with modifications. In general, PI staining is associated with damaged cell membrane or presence of dying cells (Stone et al. BMC Cell Biol. 2003, 4, 1). Due to the loss of adherence, damaged or death cells were stained by PI solution directly in media at 1.5 μM final concentration and incubated at room temperature for 15-25 minutes at room temperature. PI stained cells were quantified at 535/617 nm (excitation/emission) by Infinite M200 Pro (Tecan) microplate reader. Cell death generated by 160 mM Glu was consider as 100% of cell death so that reduction in cell death (neuroprotective effect) was observed.

[0107] Treatment by 160 mM Glu resulted in approx. 5-times higher cell death in comparison to DMSO control. As can be seen from Table 5 and FIG. 1 all tested examples at 10 μM demonstrated comparable or higher neuroprotective effect than positive controls. Specifically, the highest effect was observed in case of compounds 15 and 6 at 10 μM with double neuroprotective effect than R-LA. Additionally, compounds showed higher efficiency than positive controls due the fact that neuroprotective effect was achieved at orders of magnitude lower concentrations (5-10 fold).

TABLE-US-00003 TABLE 5 Neuroprotective effects of novel compounds on human neuroblastoma cell line SH-SY5Y (differentiated phenotype) Compound Neuroprotective effect (% of control) p value 0.5 μM 5 μM 50 μM R-LA N/A N/A 13.49 >0.001   1 μM 10 μM  100 μM  DFO N/A 19.94 23.55 >0.001 0.1 μM 1 μM 10 μM 57 N/A 15.99 24.88 >0.001 30 N/A N/A 23.17 >0.001 15 N/A N/A 30.08 >0.001 111 N/A N/A 26.52 >0.001 6 N/A 15.00 33.70 >0.001 N/A = no protection; The data are expressed as differences between 160 mM Glu and co-treatment groups (compounds) means, with significances after statistical analysis (ANOVA, Tukey post hoc test).

Example 18 Measurement of Oxidative Stress on Human Neuroblastoma Cell Line SH-SYSY-Glutamate-Induced Model of PD

[0108] Similarly as shown in previous assay, cells grew on 96 multiwell plates at density 20000 cells per well underwent differentiation and treatment procedure for glutamate model. After 4 hour co-treatment with glutamate superoxide radical's formation as marker of oxidative stress was quantified by dihydroethidium according to Kim et al. 2017, J. Med. Food, 20, 140-151 with modifications. Briefly, neuron-like SH-SYSY cells were centrifuged at 500 g for 5 minutes and 30 seconds, then media were replaced by 10 μM DHE PBS solution and kept at room temperature for 30 minutes. After 30 minutes, DHE was read at 500 nm/580 nm (excitation/emission) by Infinite M200 Pro (Tecan) microplate reader. DHE is cell permeable dye which is selective toward superoxide radical detection. Overall oxidative stress achieved by 160 mM Glu was consider as 100% so that the reduction in oxidative stress (OS reducing effect) was observed. As shown in Table 6 and FIG. 2, 160 mM Glu induced 2.6 fold increase in superoxide radical's formation. More importantly all examples at 10 μM demonstrated comparable or slightly better OS reducing effect than antioxidant R-LA, while completely outperformed DFO. Taken together, all examples showed higher efficiency than positive controls in term of concentrations responsible for OS reducing activity (5-10 fold more effective than positive controls).

TABLE-US-00004 TABLE 6 Oxidative stress-induced by glutamate and OS reducing activity of novel compounds when tested on neuron-like SH-SY5Y cells. Superoxide radical formation (oxidative stress) % of reduction average ±SEM DMSO CTR 38.18 0.74 Glu 160 mM 100 1.52 Compound 1 μM ±SEM 10 μM ±SEM 57 93.34 3.83 83.99 5.33 30 84.54 3.88 81.08 3.25 15 98.27 4.65 85.61 3.28 111 89.17 4.8 80.21 3.31 6 85.36 3.38 80.19 2.95 R-LA 5 μM ±SEM 50 μM ±SEM 86.72 5.07 83.91 4.33 DFO 10 μM  ±SEM 100 μM  ±SEM 87.06 2.39 92.61 2.2 The data are expressed as percentages of 160 mM Glu control, the mean ± SEM of three independent experiments.

Example 21—Formulations

[0109] The growth regulatory formulations usually contain from 0.1 to 99% by weight, especially from 0.1 to 95% by weight, of active ingredient mixture comprising a C2,C6-disubstituted-9-benzylated-9H-purine derivative of this invention, from 1 to 99.9% by weight of a solid or liquid formulation adjuvant, and from 0 to 25% by weight, especially from 0.1 to 25% by weight, of a surfactant. Whereas commercial products are usually formulated as concentrates, the end user will normally employ dilute formulations. The compositions may also comprise further ingredients, such as stabilizers, e.g., vegetable oils or epoxidized vegetable oils (epoxidized coconut, rapeseed oil or soybean oil), antifoams, e.g., silicone oil, preservatives, viscosity regulators, binders, tackifiers, and also fertilisers or other active ingredients. Preferred formulations have especially the following compositions: (%=percent by weight):

TABLE-US-00005 F1. Wettable powders a) b) c) d) active ingredient mixture 5% 25%  50%  80% sodium lignosulfonate 4% — 3% — sodium lauryl sulphate 2% 3% —  4% sodium diisobutylnaphthalene- — 6% 5%  6% sulfonate octylphenol polyglycol ether (7-8 mol — 1% 2% — ethylene oxide) highly dispersed silicic acid 1% 3% 5% 10% kaolin 88%  62%  35%  —

[0110] The active ingredient is mixed thoroughly with the adjuvants and the mixture is thoroughly ground in a suitable mill, affording wettable powders which can be diluted with water to give suspensions of any desired concentration.

TABLE-US-00006 F2. Suspension concentrates a) b) c) d) active ingredient mixture 3% 10%  25%  50%  ethylene glycol 5% 5% 5% 5% nonylphenol polyglycol ether (15 — 1% 2% — mol of ethylene oxide) sodium lignosulfonate 3% 3% 4% 5% carboxymethylcellulose 1% 1% 1% 1% 37% aqueous formaldehyde 0.2%.sup.  0.2%.sup.  0.2%.sup.  0.2%.sup.  solution silicone oil emulsion 0.8%.sup.  0.8%.sup.  0.8%.sup.  0.8%.sup.  water 87%  79%  62%  38% 

[0111] The finely ground active ingredient is intimately mixed with the adjutants, giving a suspension concentrate from which suspensions of any desired concentration can be obtained by dilution with water.

[0112] F3. Dry Capsules

[0113] 5000 capsules, each of which contain 0.25 g of one of the C2,C6-disubstituted-9-benzyl-9H-purine derivative as active ingredient, are prepared as follows: [0114] Composition: Active ingredient: 1250 g; Talc: 180 g; Wheat starch: 120 g; Magnesium stearate: 80 g; Lactose 20 g. [0115] Preparation process: The powdered substances mentioned are pressed through a sieve of mesh width 0.6 mm Portions of 0.33 g of the mixture are transferred to gelatine capsules with the aid of a capsule-filling machine.

[0116] F4. Soft Capsules

[0117] 5000 soft gelatine capsules, each of which contain 0.05 g of one of the C2,C6-disubstituted-9-benzyl-9H-purine derivative as active ingredient, are prepared as follows: [0118] Composition: 250 g Active ingredient+2 litres Lauroglycol [0119] Preparation process: The powdered active ingredient is suspended in Lauroglykol® (propylene glycol laurate, Gattefossé S. A., Saint Priest, France) and ground in a wet-pulveriser to a particle size of about 1 to 3 mm Portions of in each case 0.419 g of the mixture are then transferred to soft gelatine capsules by means of a capsule-filling machine.

[0120] F5. Soft Capsules

[0121] 5000 soft gelatine capsules, each of which contain 0.05 g of one of the C2,C6-disubstituted-9-benzyl-9H-purine derivative as active ingredient, are prepared as follows: [0122] Composition: 250 g Active ingredient+1 litre PEG 400+1 litre Tween 80 [0123] Preparation process: The powdered active ingredient is suspended in PEG 400 (polyethylene glycol of Mr between 380 and about 420, Sigma, Fluka, Aldrich, USA) and Tween® 80 (polyoxyethylene sorbitan monolaurate, Atlas Chem. Inc., Inc., USA, supplied by Sigma, Fluka, Aldrich, USA) and ground in a wet-pulveriser to a particle size of about 1 to 3 mm Portions of in each case 0.43 g of the mixture are then transferred to soft gelatine capsules by means of a capsule-filling machine.

Example 22—Formulation of Tablets with Controlled Release

[0124] One tablet contains, for example, 300-350 mg of terpenoid derivative as active ingredient. Excipient with known effect: Each tablet contains 150 to 200 mg of a retardant (Methocel, Parteck® SRP 80, Kollidon® SR, Kollidon 25, chitosan, alginate), as well as a lubricant (magnesium stearate), active substances (VH), binders (Prosolv SMCC 90).

[0125] The dosage form is a controlled release tablet.

[0126] Tablet preparation: Tablets are prepared by direct compression. First, the calculated amount of retarding component (Methocel, Parteck® SRP 80, Kollidon® SR, Kollidon 25, chitosan, alginate), weighing agent (magnesium stearate), active ingredient (VH), binder (Prosolv SMCC 90) are weighed. The resulting mixture is then homogenized in a homogenizer (Retsch MM200—Retsch GmbH, Haan). It is recommended to carry out the homogenization at three frequencies: 10 oscillations/s, 13 and 15 oscillations/s for 1 minute each. The tablet is then transferred to a hand press. The tablets are compressed at a load of 8 kN for 5 minutes. The load is selected with respect to the desired tablet strength of 0.8 to 0.8 MPa. The tablet weight is 500±5 mg.

[0127] Hydrophilic Matrix Tablets with Hypromellose

[0128] The tablets are prepared by the direct compression method as described above.

[0129] The tablet weight was 500±5 mg. Composition of hydrophilic tablets with hypromellose in wt. %:

TABLE-US-00007 Formulation A1 A2 A3 Prosolv SMCC 90 49% 49% 49% Methocel K4M — 30% — Methocel K15M 30% — — Methocel K100M — — 30% Active substance 20% 20% 20% Magnesium stearate  1%  1%  1%

[0130] Hydrophilic Matrix Tablets with Retarding Component Kollidon 25, Kollidon® SR, Parteck® SRP 80

[0131] The tablets are prepared by the direct compression method as described above.

[0132] The tablet weight was 500±5 mg. Composition of hydrophilic tablets in wt. %:

TABLE-US-00008 Formulation F1 F2 F3 F4 F5 Prosolv ® SMCC 90 49% 49% 49% 49% 49% Kollidon 25 30% 20% 10% — — Kollidon ® SR — — — 30% — Parteck ® SRP 80 — — — — 30% Active substance 20% 30% 40% 20% 20% Magnesium stearate  1%  1%  1%  1%  1%

[0133] Hydrophilic Matrix Tablets Containing LubriTose™ MCC, Methocel K15M Nebo Methocel K4M

[0134] The tablets are prepared by the direct compression method as described above.

[0135] The tablet weight was 500±5 mg. Composition of hydrophilic tablets in wt. %:

TABLE-US-00009 Formulation A1 A2 LubriTose ™ MCC 50% 50% Methocel K15M 30% — Methocel K4M — 30% Active substance 20% 20%