9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives and their use for the treatment of skin disorders

Abstract

The invention relates to N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2, 6-diamine derivatives and their use as drugs and cosmetics. The compounds of the present invention exhibit a number of biological activities associated with oxidative stress inhibition, especially anti-aging, anti-inflammatory and anti-neurodegenerative biological activities. The invention also relates to cosmetic and pharmaceutical compositions containing such derivatives as active agents.

Claims

1. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives of the general formula I, ##STR00002## wherein R.sup.2 and R.sup.3 are independently selected from the group consisting of H, —(CH.sub.2).sub.mCH.sub.3, m=0 or 1 or 2, —CH.sub.2(CH.sub.3).sub.2, and —(CH.sub.2).sub.n—N(CH.sub.3).sub.2, n=2 or 3, wherein at least one of R.sup.2 and R.sup.3 is alkyl or dimethylaminoalkyl; o is an integer in the range of from 2 to 5, wherein a hydrogen in at least one methylene group (CH.sub.2) of the substituent on N atom in position 9 may optionally be replaced by a methyl or methoxy group; R.sup.1 is selected from the group consisting of furfuryl or furfuryl substituted with at least one methyl or methoxy group, benzyl or benzyl substituted by at least one substituent selected from the group consisting of methyl, trifluoromethyl, hydroxy, methoxy, trifluoromethoxy, halogen, amino, methoxycarbonyl and acetoxy, 3-methylbut-2-en-1-yl, 3-methylbut-3-en-1-yl, (4-hydroxy-3-methylbut-2-en-1-yl, and 4-hydroxy-3-methylbutyl; or a pharmaceutically acceptable salt thereof with an alkali metal, ammonium or amine, in the form of a racemate or optically active isomer, or an addition salt with acid.

2. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, wherein if R.sup.2 or R.sup.3 is —(CH.sub.2).sub.nN(CH.sub.3).sub.2, the other of these substituents is hydrogen.

3. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, wherein the group —N(R.sup.2)(R.sup.3) is selected from the group consisting of methylamino, ethylamino, propylamino, 2-propylamino, dimethylamino, diethylamino, [2-(dimethylamino)ethyl]amino; and [3-(dimethylamino)propyl]amino.

4. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, wherein R1 is selected from the group consisting of furfuryl, 3-methylfurfuryl, 4-methylfurfuryl, 5-methylfurfuryl, 3-methoxyfurfuryl, 4-methoxyfurfuryl, 5-methoxyfurfuryl, benzyl, 2-methylbenzyl, 3-methylbenzyl, 4-methylbenzyl, 2-(trifluoromethyl)benzyl, 3-(trifluoromethyl)benzyl, 4-(trifluoromethyl)benzyl, 2-hydroxybenzyl, 3-hydroxybenzyl, 4-hydroxybenzylamino, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(trifluoromethoxy)benzyl, 3-(trifluoromethoxy)benzyl, 4-(trifluoromethoxy)benzyl, 2-fluorobenzyl, 3-fluorobenzyl, 4-fluorobenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2-bromobenzyl, 3-bromobenzyl, 4-bromobenzyl, 2-iodobenzyl, 3-iodobenzyl, 4-iodobenzyl, 2-aminobenzyl, 3-aminobenzyl, 4-aminobenzyl, 2-(methoxykarbonyl)benzyl, 3-(methoxykarbonyl)benzyl, 4-(methoxykarbonyl)benzyl, 2-acetoxybenzyl, 3-acetoxybenzyl, 4-acetoxybenzyl, 2,3-dihydroxybenzyl, 2,5-dihydroxybenzyl, 3,4-dihydroxybenzyl, 3,5-dihydroxybenzyl, 2,3-dimethoxybenzyl, 2,5-dimethoxybenzyl, 3,4-dimethoxybenzyl, 3,5-dimethoxybenzyl, 2,6-difluorobenzyl, 3,4-difluorobenzyl, 3,5-difluorobenzyl, 2,3-dichlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl, 3,5-dichlorobenzyl, 2-hydroxy-3-methylbenzyl, 2-hydroxy-5-methylbenzyl, 2-hydroxy-3-methoxybenzyl, 2-hydroxy-4-methoxybenzyl, 3-hydroxy-4-methoxybenzyl, 4-hydroxy-2-methoxybenzyl, 4-hydroxy-3-methoxybenzyl, 3-fluoro-4-hydroxybenzyl, 3-chloro-4-hydroxybenzyl, 4-fluoro-3-hydroxybenzyl, 4-chloro-3-hydroxybenzyl, 2-chloro-4-fluorobenzyl, 2-chloro-6-fluorobenzyl, 3,4,5-trihydroxybenzyl, 3,4,5-trimethoxybenzyl, 2,3,4-trifluorobenzyl, 2,3,6-trifluorobenzyl, 3,4,5-trifluorobenzyl, 4-hydroxy-3,5-dimethoxybenzyl, 3-methylbut-2-en-1-yl, 3-methylbut-3-en-1-yl, 4-hydroxy-3-methylbut-2-en-1-yl, and 4-hydroxy-3-methylbutyl.

5. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, wherein the substituent on N atom in position 9 is selected from tetrahydro-2H-pyran-2-yl and tetrahydrofuran-2-yl, which may optionally be substituted by at least one methyl or methoxy group.

6. The N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, selected from the group consisting of 2-(methylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine, 2-(ethylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine; 2-(dimethylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine; 2-(diethylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine; 2-{[2-(dimethylamino)ethyl]amino}-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine; 2-(methylamino)-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine, 2-(ethylamino)-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine; 2-(dimethylamino)-6-furfurylamino-9-(tetrahydro-2N-pyran-2-yl)-9H-purine; 2-(diethylamino)-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine; and 2-{[2-(dimethylamino)ethyl]amino}-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine, or a salt thereof, wherein the furfuryl group can optionally be substituted by at least one substituent selected from the group consisting of methyl and methoxy group.

7. A method for inhibiting lipid and protein peroxidation and/or inhibiting aging processes comprising the step of providing N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives of the general formula I, ##STR00003## wherein R.sup.2 and R.sup.3 are independently selected from the group consisting of H, —(CH.sub.2).sub.mCH.sub.3, m=0 or 1 or 2, —CH.sub.2(CH.sub.3).sub.2, and —(CH.sub.2).sub.n—N(CH.sub.3).sub.2, n=2 or 3, wherein at least one of R.sup.2 and R.sup.3 is alkyl or dimethylaminoalkyl; o is an integer in the range of from 2 to 5, wherein a hydrogen in at least one methylene group (CH.sub.2) of the substituent on N atom in position 9 may optionally be replaced by a methyl or methoxy group; R.sup.1 is selected from the group consisting of furfuryl or furfuryl substituted with at least one methyl or methoxy group, benzyl or benzyl substituted by at least one substituent selected from the group consisting of methyl, trifluoromethyl, hydroxy, methoxy, trifluoromethoxy, halogen, amino, methoxycarbonyl and acetoxy, 3-methylbut-2-en-1-yl, 3-methylbut-3-en-1-yl, (4-hydroxy-3-methylbut-2-en-1-yl, and 4-hydroxy-3-methylbutyl; or a pharmaceutically acceptable salt thereof with an alkali metal, ammonium or amine, in the form of a racemate or optically active isomer, or an addition salt with acid to a subject in need thereof.

8. The method according to claim 7, wherein the method comprises treatment of psoriasis or multiple sclerosis.

9. The method according to claim 7, wherein the method comprises stimulation of proliferation, morphogenesis and senescence inhibition, cell division and differentiation factors of plant, mammal, microorganisms, yeast and fungal cells.

10. Pharmaceutical composition, characterized in that it comprises at least one N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative of the general formula I according to claim 1, and at least one auxiliary substance.

11. A cosmetic composition, characterized in that it comprises at least one N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative of the general formula I according to claim 1, and at least one auxiliary substance.

12. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives according to claim 1, wherein the substituent on N atom in position 9 is selected from tetrahydro-2H-pyran-2-yl and tetrahydrofuran-2-yl, which may optionally be substituted by one methyl or methoxy group.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 shows inhibition of intracellular ROS production by compound 114. The bars show mean±SD. Differences between control cells and cells treated with compound 114 were tested by Student t-test (** p<0.01, *** p<0.001).

EXAMPLES

(2) 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.

(3) The starting materials may be obtained from commercial sources (Sigma, Aldrich, Fluka, etc.) or can be prepared as described below. Thin-layer chromatography was carried out on Silica 60 F.sub.254 plates (Merck) using CHCl.sub.3/MeOH as a developing system and the spots were detected by UV light (254 and 365 nm) and/or 6% (v/v) vanilline in absolute EtOH containing 1% (v/v) of H.sub.2SO.sub.4. The column chromatography purification of intermediates was carried out by silica Davisil 40-63 micron (Grace Davision). Elemental analysis was determined using Flash EA 1112 analyzer (Thermo Scientific). The chromatographic purity and molecular mass of prepared compounds was determined using an Alliance 2695 separation module (Waters) linked simultaneously to a DAD detector PDA 996 (Waters) and a Q-Tof micro (Waters) benchtop quadrupole orthogonal acceleration time-of-flight tandem mass spectrometer. Samples were dissolved in DMSO and diluted to a concentration of 10 μg/ml in initial mobile phase (90% 15 mM ammonium formate, pH 4.0 (A)+10% methanol (B)). Samples (10 μl) were injected on a RP-column Symmetry C18 (150 mm×2.1 mm×3.5 μm, Waters) and separated at a flow rate of 0.2 ml/min with the following binary gradient: 0 min, 10% B; 0-24 min, a linear gradient to 90% B, 10 min, followed by 10 min isocratic elution of 90% B. At the end of the gradient, the column was re-equilibrated to initial conditions (15 mM formic acid adjusted to pH 4.0 by ammonium hydroxide was used as solvent (A) and methanol as the organic modifier (solvent B)). The eluent was introduced into the DAD (scanning range 210-400 nm, with 1.2 nm resolution) and an ESI source (source temperature 110° C., capillary voltage +3.0 kV, cone voltage +20 V, desolvation temperature 250° C.). Nitrogen was used both as desolvation gas (500 l/h) as well as cone gas (50 l/h). The data was obtained in positive (ESI+) ionization mode in the 50-1000 m/z range.

(4) .sup.1H and .sup.13C NMR spectra were recorded on Jeol ECA-500 operating at a frequency of 500 MHz (.sup.1H) and 126 MHz (.sup.13C) or Bruker Avance operating at a frequency of 300 MHz (.sup.1H) and 75 MHz (.sup.13C), respectively. Samples were prepared by dissolving substances in DMSO-d.sub.6 and chemical shifts were calibrated to residual solvent peak (DMSO, 2.49 ppm for proton) and DMSO-d.sub.6 (39.5 ppm for carbon).

(5) N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivatives were prepared in three steps, starting from 2,6-dichloro-9H-purine. Firstly, (2-oxacycloalkyl) substituents were introduced to the N9-position of purine moiety by hydroamination of corresponding α,β-unsaturated O-heterocycles by 2,6-dichloro-9H-purine in ethyl acetate catalyzed by trifluoroacetic acid according to protocol in Szučová et al. 2009 Bioorg. Med. Chem. 17, 1938-1947. 2,3,4,5-tetrahydrooxepin was prepared by endo cycloisomeration of 5-hexyn-1-ol catalyzed by tetradentate nitrogen-phosphorous mixed ligand ruthenium complex following methodology published in Liu et al. (2010) Chem. Eur. J. 16, 7889-7897 and Mitchell et al. (1973) J. Chem. Soc., Dalton Trans. (8), 846-854.

(6) The intermediates were further subjected to the nucleophilic substitution at C6 position with appropriate side-chain amines and triethylamine in refluxing propanol followed by nucleophilic substitution of C2 chlorine atom. Small amines such as methylamine or dimethylamine were introduced to C2 position using a modified protocol published in Kelley et al. (1989) J. Med. Chem. 32, 218-224, whereas amines with larger molecular mass were introduced to C2 position according to Kurimoto et al. (2003) Bioorg. Med. Chem. 11, 5501-5508.

Modification of Purines at N9-Position

(7) A) Hydroamination of α,β-Unsaturated O-Heterocycles

Example 1 2,6-Dichloro-9-(tetrahydrofuran-2-yl)-9H-purine

(8) To a suspension of 2,6-dichloro-9H-purine (1 eq., 5 g) and 2,3-dihydrofuran (2.5 eq.) under Ar atmosphere in dry ethyl acetate (40 ml) was trifluoroacetic acid (2.5 eq) dropwise added and resulting solution was stirred at room temperature for 2 hours. After cooling in ice bath pH of the reaction mixture was adjusted to 7-8 by conc. NH.sub.3/water (2:3, v/v) and the layers were separated. The water phase was re-extracted by ethyl acetate (5×25 ml). Combined organic layers were washed with brine (2×25 ml), dried (Na.sub.2SO.sub.4) and evaporated under reduced pressure. The product as a pale yellow solid was obtained by crystallization from petroleum ether. Pale yellow solid, chemical formula: C.sub.9H.sub.8Cl.sub.2N.sub.4O, yield (%): 98. HPLC-UV/VIS retention time, purity (min., %): 21.07, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 258.8 (8, [.sup.35Cl-M+H].sup.+), 260.8 (6, [.sup.37Cl-M+H].sup.+), 188.8 (100, [.sup.35Cl-M-THF+H].sup.+), 190.8 (68, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 2.01-2.08 (m, 1H), 2.11-2.20 (m, 1H), 2.41-2.46 (m, 2H), 3.93 (q, J=7.5 Hz, 1H), 4.18 (td, J=7.9, 5.7 Hz, 1H), 6.33 (dd, J=5.8, 4.3 Hz, 1H), 8.82 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 23.8, 31.3, 69.2, 85.6, 131.0, 146.4, 149.6, 150.9, 152.6.

Example 2 2,6-Dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(9) To a suspension of 2,6-dichloro-9H-purine (1 eq., 5 g) and 3,4-dihydro-2H-pyran (2 eq.) under Ar atmosphere in dry ethyl acetate (50 ml) was trifluoroacetic acid (1.8 eq.) dropwise added and resulting solution was stirred at room temperature for 2 hours. After cooling in ice bath pH of the reaction mixture was adjusted to 7-8 by conc. NH.sub.3/water (2:3, v/v). Resulting solid was filtered, washed with cold water and dried at room temperature. Second portion of product was obtained by extraction of filtrate and washings by ethyl acetate (3×25 ml). The combined organic layers were washed with brine (2×15 ml) dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure to give a yellow gel. The solid material was obtained after crystallization from diethyl ether. White solid, chemical formula: C.sub.10H.sub.10Cl.sub.2N.sub.4O, yield (%): 88. HPLC-UV/VIS retention time, purity (min., %): 22.87, 98.4. ESI.sup.+-MS m/z (rel. int. %, ion): 273.0 (10, [.sup.35Cl-M+H].sup.+), 275.0 (8, [.sup.37Cl-M+H].sup.+), 189.0 (100, [.sup.35Cl-M-THP+H].sup.+), 191.0 (74, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.56-1.61 (m, 2H), 1.73-1.77 (m, 1H), 1.94-2.01 (m, 2H), 2.21-2.29 (m, 1H), 3.70-3.75 (m, 1H), 4.00-4.03 (m, 1H), 5.73 (dd, J=10.9, 2.3 Hz, 1H), 8.95 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.0, 24.3, 29.6, 67.7, 81.6, 130.5, 146.4, 149.9, 151.2, 152.7.

Substitution of Purines at C6 Position

(10) Synthesis of 2-chloro-6-substituted-9-(2-oxacycloalkyl)-9H-purine Derivatives—Typical Procedure

(11) Mixture of 2,6-dichloro-9-(2-oxacycloalkyl)-9H-purine (1 eq., 250 mg), side-chain amine (1.2 eq.) and triethylamine (2.5 eq. and in case of salts 3.5 eq., respectively) were refluxed in propanol (9 ml) for 4 hours. After evaporation under reduced pressure the residue was treated with ice cold water (15 ml).

(12) Method A) If a solid material was obtained—the solid was filtered, washed well with ice cold water and dried.

(13) Method B) If no solid material was obtained—the mixture was extracted by ethyl acetate (5×10 ml). Combined organic layers were washed using brine (2×10 ml), dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure.

(14) The crude material was purified by silica column chromatography using chloroform/methanol as a mobile phase starting from pure chloroform with methanol gradient.

Example 3 2-Chloro-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(15) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and furfurylamine, work-up using A. White solid, chemical formula: C.sub.14H.sub.14ClN.sub.5O.sub.2, yield (%): 85. HPLC-UV/VIS retention time, purity (min., %): 24.43, 99.8. ESI.sup.+-MS m/z (rel. int. %, ion): 319.8 (100, [.sup.35Cl-M+H].sup.+), 321.8 (45, [.sup.37Cl-M+H].sup.+), 249.8 (83, [.sup.35Cl-M-THF+H].sup.+), 251.8 (35, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.96-2.04 (m, 1H), 2.11-2.20 (m, 1H), 2.36-2.41 (m, 2H), 3.89 (q, J=7.3 Hz, 1H), 4.11 (td, J=7.8, 5.9 Hz, 1H), 4.61 (d, J=5.5 Hz, 2H), 6.19 (t, J=5.2 Hz, 1H), 6.25 (d, J=2.8 Hz, 1H), 6.37 (bs, 1H), 7.54-7.55 (m, 1H), 8.28 (s, 1H), 8.76 (t, J=5.2 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.1, 31.2, 36.6, 68.7, 84.4, 107.1, 110.5, 118.6, 139.7, 142.0, 149.3, 152.0, 152.9, 154.7.

Example 4 2-Chloro-6-[(5-methylfurfuryl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(16) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 5-methylfurfurylamine, work-up using B. White solid, chemical formula: C.sub.15H.sub.16ClN.sub.5O.sub.2, yield (%): 66. HPLC-UV/VIS retention time, purity (min., %): 26.05, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 334.6 (100, [.sup.35Cl-M+H].sup.+), 336.7 (64, [.sup.37Cl-M+H].sup.+), 264.5 (87, [.sup.35Cl-M-THF+H].sup.+), 266.6 (43, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.96-2.04 (m, 1H), 2.13-2.17 (m, 1H), 2.20 (s, 3H), 2.36-2.41 (m, 2H), 3.89 (q, J=7.3 Hz, 1H), 4.11 (q, J=7.2 Hz, 1H), 4.55 (d, J=5.5 Hz, 2H), 5.96 (bs, 1H), 6.11 (d, J=2.8 Hz, 1H), 6.19 (t, J=5.2 Hz, 1H), 8.28 (s, 1H), 8.72 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 13.3, 24.1, 31.2, 36.6, 68.7, 84.4, 106.4, 107.9, 118.6, 139.6, 149.3, 150.1, 150.5, 152.9, 154.6.

Example 5 2-Chloro-6-benzylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(17) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and benzylamine, work-up using B. White solid, chemical formula: C.sub.16H.sub.16ClN.sub.5O, yield (%): 65. HPLC-UV/VIS retention time, purity (min., %): 26.39, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 330.7 (100, [.sup.35Cl-M+H].sup.+), 332.7 (63, [.sup.37Cl-M+H].sup.+), 260.5 (82, [.sup.35Cl-M-THF+H].sup.+), 262.6 (40, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.99-2.04 (m, 1H), 2.13-2.18 (m, 1H), 2.38-2.39 (m, 2H), 3.88-3.90 (m, 1H), 4.10-4.11 (m, 1H), 4.63 (bs, 2H), 6.18-6.19 (m, 1H), 7.22 (bs, 1H), 7.31 (bs, 4H), 8.28 (s, 1H), 8.86 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.1, 31.2, 43.1, 68.7, 84.4, 118.5, 126.8, 2×127.2, 2×128.2, 139.2, 139.6, 149.2, 153.0, 154.9.

Example 6 2-Chloro-6-[(3-hydroxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(18) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 3-(aminomethyl)phenol, work-up using A. White solid, chemical formula: C.sub.16H.sub.16ClN.sub.5O.sub.2, yield (%): 94. HPLC-UV/VIS retention time, purity (min., %): 23.33, 99.2. ESI.sup.+-MS m/z (rel. int. %, ion): 345.8 (100, [.sup.35Cl-M+H].sup.+), 347.8 (40, [.sup.37Cl-M+H].sup.+), 275.8 (83, [.sup.35Cl-M-THF+H].sup.+), 277.8 (35, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.98-2.03 (m, 1H), 2.14-2.19 (m, 1H), 2.39 (dd, J=12.8, 7.3 Hz, 2H), 3.89 (q, J=7.3 Hz, 1H), 4.11 (dd, J=13.8, 7.6 Hz, 1H), 4.56 (d, J=6.1 Hz, 2H), 6.19 (t, J=5.2 Hz, 1H), 6.60 (dd, J=7.9, 1.5 Hz, 1H), 6.70 (s, 1H), 6.73 (d, J=7.8 Hz, 1H), 7.08 (t, J=7.6 Hz, 1H), 8.28 (s, 1H), 8.81 (t, J=6.1 Hz, 1H), 9.31 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.1, 31.2, 42.9, 68.7, 84.4, 113.7, 113.8, 117.7, 118.5, 129.2, 139.5, 140.7, 149.2, 153.1, 154.9, 157.3.

Example 7 2-Chloro-6-[(4-hydroxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(19) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 4-(aminomethyl)phenol, work-up using A. White solid, chemical formula: C.sub.16H.sub.16ClN.sub.5O.sub.2, yield (%): 98. HPLC-UV/VIS retention time, purity (min., %): 23.02, 97.6. ESI.sup.+-MS m/z (rel. int. %, ion): 345.8 (100, [.sup.35Cl-M+H].sup.+), 347.8 (45, [.sup.37Cl-M+H].sup.+), 275.8 (85, [.sup.35Cl-M-THF+H].sup.+), 277.8 (35, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.96-2.04 (m, 1H), 2.12-2.20 (m, 1H), 2.36-2.41 (m, 2H), 3.89 (q, J=7.3 Hz, 1H), 4.11 (td, J=7.7, 6.2 Hz, 1H), 4.51 (d, J=5.2 Hz, 2H), 6.18 (t, J=5.2 Hz, 1H), 6.68 (d, J=8.6 Hz, 2H), 7.14 (d, J=8.3 Hz, 2H), 8.25 (s, 1H), 8.69 (app. t, 1H), 9.23 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.0, 31.1, 42.6, 68.6, 84.3, 2×114.9, 118.5, 2×128.6, 129.3, 139.3, 149.1, 153.0, 154.7, 156.2.

Example 8 2-Chloro-6-[(2-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(20) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 2-methoxybenzylamine, work-up using B. White solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O.sub.2, yield (%): 53. HPLC-UV/VIS retention time, purity (min., %): 26.97, 98.7. ESI.sup.+-MS m/z (rel. int. %, ion): 360.3 (100, [.sup.35Cl-M+H].sup.+), 362.3 (65, [.sup.37Cl-M+H].sup.+), 290.6 (88, [.sup.35Cl-M-THF+H].sup.+), 292.6 (46, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.99-2.03 (m, 1H), 2.14-2.18 (m, 1H), 2.38-2.42 (m, 2H), 3.82 (s, 3H), 3.89 (q, J=7.3 Hz, 1H), 4.09-4.14 (m, 1H), 4.61 (d, J=6.1 Hz, 2H), 6.19 (t, J=5.0 Hz, 1H), 6.85 (t, J=7.5 Hz, 1H), 6.98 (d, J=7.9 Hz, 1H), 7.10 (d, J=7.3 Hz, 1H), 7.21 (t, J=7.6 Hz, 1H), 8.30 (s, 1H), 8.63 (t, J=6.0 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.1, 31.2, 38.4, 55.3, 68.7, 84.4, 110.4, 118.6, 120.1, 126.43, 126.8, 127.9, 139.5, 149.2, 153.1, 155.1, 156.5.

Example 9 2-Chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(21) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 4-methoxybenzylamine, work-up using B. Pale yellow solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O.sub.2, yield (%): 54. HPLC-UV/VIS retention time, purity (min., %): 26.17, 98.3. ESI.sup.+-MS m/z (rel. int. %, ion): 360.3 (100, [35Cl-M+H].sup.+), 362.3 (68, [37Cl-M+H].sup.+), 290.6 (87, [.sup.35Cl-M-THF+H].sup.+), 292.6 (48, [37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.97-2.02 (m, 1H), 2.13-2.18 (m, 1H), 2.36-2.40 (m, 2H), 3.70 (s, 3H), 3.88 (q, J=7.4 Hz, 1H), 4.08-4.12 (m, 1H), 4.55 (d, J=6.1 Hz, 2H), 6.18 (t, J=5.2 Hz, 1H), 6.86 (d, J=8.6 Hz, 2H), 7.26 (d, J=8.9 Hz, 2H), 8.27 (s, 1H), 8.79 (t, J=6.0 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.1, 31.2, 42.5, 55.0, 68.7, 84.4, 2×113.6, 118.5, 128.4, 128.7, 131.1, 139.5, 149.1, 153.0, 154.7, 158.2.

Example 10 2-Chloro-6-[(4-hydroxy-3-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(22) Prepared from 2,6-dichloro-9-(tetrahydrofuran-2-yl)-9H-purine and 4-(aminomethyl)-2-methoxyphenol hydrochloride, work-up using A. White solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O.sub.3, yield (%): 96. HPLC-UV/VIS retention time, purity (min., %): 23.27, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 375.8 (99, [.sup.35Cl-M+H].sup.+), 377.8 (43, [.sup.37Cl-M+H].sup.+), 305.8 (100, [.sup.35Cl-M-THF+H].sup.+), 307.8 (42, [.sup.37Cl-M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.96-2.04 (m, 1H), 2.12-2.20 (m, 1H), 2.36-2.41 (m, 2H), 3.73 (s, 3H), 3.89 (q, J=7.3 Hz, 1H), 4.11 (td, J=7.7, 6.2 Hz, 1H), 4.51 (d, J=5.5 Hz, 2H), 6.18 (t, J=5.2 Hz, 1H), 6.68 (d, J=7.9 Hz, 1H), 6.73 (d, J=7.9 Hz, 1H), 7.00 (s, 1H), 8.25 (s, 1H), 8.68 (app. t, 1H), 8.79 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.0, 31.1, 43.0, 55.4, 68.6, 84.3, 112.3, 115.1, 118.5, 120.01, 129.9, 139.3, 145.4, 147.2, 149.1, 153.0, 154.7.

Example 11 2-Chloro-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(23) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and furfurylamine, work-up using A. White solid, chemical formula: C.sub.15H.sub.16ClN.sub.5O.sub.2, yield (%): 83. HPLC-UV/VIS retention time, purity (min., %): 25.57, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 334.0 (100, [.sup.35Cl-M+H].sup.+), 336.0 (56, [.sup.37Cl-M+H].sup.+), 249.9 (92, [.sup.35Cl-M-THP+H].sup.+), 251.9 (44, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.58 (m, 2H), 1.68-1.74 (m, 1H), 1.91-1.95 (m, 2H), 2.15-2.23 (m, 1H), 3.65-3.70 (m, 1H), 3.97-3.99 (m, 1H), 4.62 (d, J=4.0 Hz, 2H), 5.55-5.57 (m, 1H), 6.26 (d, J=2.8 Hz, 1H), 6.37 (bs, 1H), 7.55 (d, J=0.9 Hz, 1H), 8.39 (s, 1H), 8.78 (t, J=5.3 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.2, 24.4, 29.9, 36.6, 67.6, 80.8, 107.0, 110.4, 118.1, 139.51, 141.9, 149.4, 151.9, 153.1, 154.6.

Example 12 2-Chloro-6-[(5-methylfurfuryl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(24) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5-methylfurfurylamine, work-up using B. White solid, chemical formula: C.sub.16H.sub.18ClN.sub.5O.sub.2, yield (%): 96. HPLC-UV/VIS retention time, purity (min., %): 26.98, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 348.8 (100, [.sup.35Cl-M+H].sup.+), 350.8 (64, [.sup.37Cl-M+H].sup.+), 264.7 (91, [.sup.35Cl-M-THP+H].sup.+), 266.7 (47, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.58 (m, 2H), 1.67-1.76 (m, 1H), 1.91-1.95 (m, 2H), 2.13-2.20 (m, 4H), 3.65-3.70 (m, 1H), 3.98 (dd, J=10.9, 1.7 Hz, 1H), 4.55 (d, J=5.5 Hz, 2H), 5.55-5.57 (m, 1H), 5.96 (bs, 1H), 6.11 (d, J=3.1 Hz, 1H), 8.38 (s, 1H), 8.73 (t, J=5.5 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 13.2, 22.3, 24.4, 29.9, 36.7, 67.6, 80.8, 106.4, 107.9, 118.1, 139.5, 149.4, 150.1, 150.5, 153.1, 154.6.

Example 13 2-Chloro-6-benzylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(25) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and benzylamine, work-up using A. White solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O, yield (%): 97. HPLC-UV/VIS retention time, purity (min., %): 27.28, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 344.8 (100, [.sup.35Cl-M+H].sup.+), 346.8 (64, [.sup.37Cl-M+H].sup.+), 260.6 (90, [.sup.35Cl-M-THP+H].sup.+), 262.6 (47, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.60 (m, 2H), 1.67-1.76 (m, 1H), 1.93 (bs, 2H), 2.16-2.24 (m, 1H), 3.65-3.70 (m, 1H), 3.98 (d, J=11.0 Hz, 1H), 4.64 (d, J=3.7 Hz, 2H), 5.55 (dd, J=11.0, 1.8 Hz, 1H), 7.22 (t, J=6.6 Hz, 1H), 7.28-7.33 (m, 4H), 8.38 (s, 1H), 8.88 (t, J=6.0 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.3, 24.4, 29.9, 43.1, 67.6, 80.8, 118.0, 126.8, 2×127.2, 2×128.2, 139.2, 139.4, 149.3, 153.2, 154.9.

Example 14 2-Chloro-6-[(4-methylbenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(26) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 4-methylbenzylamine, work-up using B. White solid, chemical formula: C.sub.18H.sub.20ClN.sub.5O, yield (%): 99. HPLC-UV/VIS retention time, purity (min., %): 28.63, 98.5. ESI.sup.+-MS m/z (rel. int. %, ion): 358.8 (100, [.sup.35Cl-M+H].sup.+), 360.8 (72, [.sup.37Cl-M+H].sup.+), 274.6 (90, [.sup.35Cl-M-THP+H].sup.+), 276.6 (59, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.58 (m, 2H), 1.67-1.76 (m, 1H), 1.90-1.94 (m, 2H), 2.15-2.27 (m, 4H), 3.64-3.70 (m, 1H), 3.98 (d, J=11.3 Hz, 1H), 4.58 (d, J=4.9 Hz, 2H), 5.55 (dd, J=10.9, 2.0 Hz, 1H), 7.10 (d, J=7.6 Hz, 2H), 7.20 (d, J=7.9 Hz, 2H), 8.37 (s, 1H), 8.84 (t, J=6.1 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 20.6, 22.3, 24.5, 29.9, 42.8, 67.6, 80.8, 118.0, 2×127.2, 2×128.8, 135.8, 136.1, 139.4, 149.3, 153.3, 154.9.

Example 15 2-Chloro-6-{[4-(trifluoromethyl)benzyl]amino}-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(27) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 4-(trifluoromethyl)benzylamine, work-up using A. White solid, chemical formula: C.sub.18H.sub.17ClF.sub.3N.sub.5O, yield (%): 73. HPLC-UV/VIS retention time, purity (min., %): 29.12, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 412.7 (97, [.sup.35Cl-M+H].sup.+), 414.7 (65, [.sup.37Cl-M+H].sup.+), 328.7 (90, [.sup.35Cl-M-THP+H]), 330.7 (64, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.55 (bs, 2H), 1.67-1.76 (m, 1H), 1.93 (bs, 2H), 2.16-2.24 (m, 1H), 3.65-3.70 (m, 1H), 3.98 (d, J=11.3 Hz, 1H), 4.72 (d, J=5.5 Hz, 2H), 5.55-5.57 (m, 1H), 7.53 (d, J=7.9 Hz, 2H), 7.68 (d, J=8.3 Hz, 2H), 8.41 (s, 1H), 8.98 (t, J=6.0 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.3, 24.4, 29.9, 42.8, 67.6, 80.9, 118.1, 124.3 (q, .sup.1J.sub.CF=272.3 Hz), 2×125.2 (q, .sup.3J.sub.CF=2.4 Hz), 127.5 (q, .sup.2J.sub.CF=32.2 Hz), 2×127.8, 139.7, 144.1, 149.4, 153.2, 154.9.

Example 16 2-Chloro-6-[(3-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(28) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 3-(aminomethyl)phenol, work-up using B. White solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O.sub.2, yield (%): 73. HPLC-UV/VIS retention time, purity (min., %): 24.43, 99.8. ESI.sup.+-MS m/z (rel. int. %, ion): 359.8 (100, [.sup.35Cl-M+H].sup.+), 361.8 (40, [.sup.37Cl-M+H].sup.+), 275.8 (99, [.sup.35Cl-M-THP+H].sup.+), 277.8 (38, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.58 (m, 2H), 1.70-1.74 (m, 1H), 1.90-1.95 (m, 2H), 2.16-2.21 (m, 1H), 3.65-3.70 (m, 1H), 3.96-3.99 (m, 1H), 4.56 (d, J=6.1 Hz, 2H), 5.56 (dd, J=11.0, 2.1 Hz, 1H), 6.60 (dd, J=7.9, 1.8 Hz, 1H), 6.70 (s, 1H), 6.73 (d, J=7.9 Hz, 1H), 7.08 (t, J=7.8 Hz, 1H), 8.38 (s, 1H), 8.83 (t, J=6.3 Hz, 1H), 9.31 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.3, 24.5, 29.9, 42.9, 67.7, 80.9, 113.7, 113.8, 117.7, 118.06, 129.2, 139.4, 140.6, 149.3, 153.3, 154.9, 157.3.

Example 17 2-Chloro-6-[(3-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(29) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 4-(aminomethyl)phenol, work-up using B. White solid, chemical formula: C.sub.17H.sub.18ClN.sub.5O.sub.2, yield (%): 99. HPLC-UV/VIS retention time, purity (min., %): 23.98, 98.0. ESI.sup.+-MS m/z (rel. int. %, ion): 360.8 (90, [.sup.35Cl-M+H].sup.+), 362.8 (51, [.sup.37Cl-M+H].sup.+), 276.7 (100, [.sup.35Cl-M-THP+H].sup.+), 278.6 (58, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.57 (m, 2H), 1.67-1.76 (m, 1H), 1.90-1.94 (m, 2H), 2.15-2.23 (m, 1H), 3.64-3.70 (m, 1H), 3.98 (d, J=11.3 Hz, 1H), 4.51 (d, J=5.8 Hz, 2H), 5.55 (d, J=10.7 Hz, 1H), 6.68 (d, J=7.9 Hz, 2H), 7.13 (d, J=8.3 Hz, 2H), 8.36 (s, 1H), 8.76 (t, J=6.1 Hz, 1H), 9.27 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.3, 24.4, 29.9, 42.6, 67.6, 80.8, 2×115.0, 118.0, 128.5, 128.7, 129.3, 139.3, 149.2, 153.3, 154.8, 156.3.

Example 18 2-Chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(30) Prepared from 2,6-dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 4-methoxybenzylamine, work-up using B. White solid, chemical formula: C.sub.18H.sub.20ClN.sub.5O.sub.2, yield (%): 99. HPLC-UV/VIS retention time, purity (min., %): 27.10, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 374.8 (98, [.sup.35Cl-M+H].sup.+), 376.8 (65, [.sup.37Cl-M+H].sup.+), 290.7 (100, [.sup.35Cl-M-THP+H].sup.+), 292.7 (62, [.sup.37Cl-M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.53-1.60 (m, 2H), 1.67-1.76 (m, 1H), 1.90-1.95 (m, 2H), 2.15-2.23 (m, 1H), 3.64-3.67 (m, 1H), 3.70 (s, 3H), 3.98 (dd, J=10.9, 1.7 Hz, 1H), 4.55 (d, J=4.6 Hz, 2H), 5.55 (dd, J=10.9, 1.7 Hz, 1H), 6.86 (d, J=8.6 Hz, 2H), 7.26 (d, J=8.9 Hz, 2H), 8.37 (s, 1H), 8.82 (t, J=6.1 Hz, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.3, 24.4, 29.9, 42.6, 55.0, 67.6, 80.8, 2×113.6, 118.0, 128.4, 128.7, 131.1, 139.4, 149.2, 153.2, 154.8, 158.2.

(31) Substitution of Purines at C2 Position:

(32) A) Substitution with Primary and Secondary Alkylamines—Typical Procedure

(33) A mixture of 2-chloro-6-substituted-9-(2-oxacycloalkyl)-9H-purine (150 mg, 1 eq.) and amine (20 eq.) was heated at 95° C. under Ar atmosphere in ACE pressure tube for 16 hours. The reaction mixture was evaporated under reduced pressure and the residue was treated with ice cold water (5 ml). The resulting solid was filtered, washed with ice cold water and dried at ambient temperature. The crude material was purified by silica column chromatography using chloroform/methanol as a mobile phase starting from pure chloroform with methanol gradient.

Example 19 2-(Methylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(34) Prepared from 2-chloro-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.15H.sub.18N.sub.6O.sub.2, yield (%): 74. HPLC-UV/VIS retention time, purity (min., %): 22.03, 99.8. ESI.sup.+-MS m/z (rel. int. %, ion): 315.0 (100, [M+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.93-2.01 (m, 1H), 2.18-2.26 (m, 1H), 2.29-2.36 (m, 1H), 2.41 (bs, 1H), 2.76 (d, J=4.9 Hz, 3H), 3.85 (q, J=7.5 Hz, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.60 (bs, 2H), 6.07 (dd, J=6.7, 3.7 Hz, 1H), 6.22 (d, J=2.8 Hz, 1H), 6.34 (m, 2H), 7.51 (s, 1H), 7.67 (bs, 1H), 7.81 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.5, 28.1, 30.7, 36.6, 68.3, 83.6, 106.5, 110.3, 113.5, 135.55, 141.5, 150.8, 153.4, 154.2, 159.7.

Example 20 2-(Dimethylamino)-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(35) Prepared from 2-chloro-6-furfurylamino-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.16H.sub.20N.sub.6O.sub.2, yield (%): 97. HPLC-UV/VIS retention time, purity (min., %): 27.07, 99.3. ESI.sup.+-MS m/z (rel. int. %, ion): 328.8 (100, [M+H].sup.+), 258.8 ([M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-2.01 (m, 1H), 2.21-2.27 (m, 1H), 2.30-2.37 (m, 1H), 2.43-2.48 (m, 1H), 3.06 (s, 6H), 3.84-3.88 (m, 1H), 4.10 (q, J=7.3 Hz, 1H), 4.59 (bs, 2H), 6.09 (dd, J=7.3, 4.0 Hz, 1H), 6.21 (d, J=2.8 Hz, 1H), 6.34 (dd, J=3.1, 1.8 Hz, 1H), 7.51 (d, J=1.2 Hz, 1H), 7.79 (bs, 1H), 7.83 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.7, 30.5, 36.3, 2×36.9, 68.5, 83.8, 106.5, 110.4, 113.1, 136.27, 141.6, 150.8, 153.5, 153.7, 159.0.

Example 21 2-(Dimethylamino)-6-[(5-methylfurfuryl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(36) Prepared from 2-chloro-6-[(5-methylfurfuryl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.17H.sub.22N.sub.6O.sub.2, yield (%): 66. HPLC-UV/VIS retention time, purity (min., %): 28.55, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 343.7 (100, [M+H].sup.+), 273.6 (70, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-2.00 (m, 1H), 2.19 (s, 3H), 2.22-2.27 (m, 1H), 2.30-2.37 (m, 1H), 2.43-2.47 (m, 1H), 3.07 (s, 6H), 3.86 (dd, J=13.4, 7.6 Hz, 1H), 4.10 (q, J=7.3 Hz, 1H), 4.52 (bs, 2H), 5.93 (d, J=1.8 Hz, 1H), 6.07-6.10 (m, 2H), 7.71 (bs, 1H), 7.82 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 13.3, 24.7, 30.5, 36.3, 2×36.9, 68.5, 83.8, 106.3, 107.3, 113.1, 136.2, 150.0, 150.7, 151.6, 153.6, 159.0.

Example 22 2-(Methylamino)-6-benzylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(37) Prepared from 2-chloro-6-benzylamino-9-(tetrahydrofuran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.17H.sub.20N.sub.6O, yield (%): 58. HPLC-UV/VIS retention time, purity (min., %): 24.14, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 325.7 (100, [M+H].sup.+), 255.6 (38, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.93-2.01 (m, 1H), 2.19-2.26 (m, 1H), 2.30-2.36 (m, 1H), 2.42 (bs, 1H), 2.73 (d, J=4.9 Hz, 3H), 3.85 (dd, J=13.9, 7.5 Hz, 1H), 4.10 (q, J=7.3 Hz, 1H), 4.61 (bs, 2H), 6.07 (dd, J=6.6, 3.8 Hz, 1H), 6.31 (bs, 1H), 7.18 (t, J=7.2 Hz, 1H), 7.27 (t, J=7.6 Hz, 2H), 7.33 (d, J=7.6 Hz, 2H), 7.80 (s, 2H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.6, 28.2, 30.7, 42.6, 68.4, 83.6, 113.5, 126.4, 2×127.3, 2×128.0, 135.5, 140.7, 150.6, 154.4, 159.8.

Example 23 2-(Dimethylamino)-6-benzylamino-9-(tetrahydrofuran-2-yl)-9H-purine

(38) Prepared from 2-chloro-6-benzylamino-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.18H.sub.22N.sub.6O, yield (%): 74. HPLC-UV/VIS retention time, purity (min., %): 28.60, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 339.7 (100, [M+H].sup.+), 269.6 (73, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-1.99 (m, 1H), 2.21-2.26 (m, 1H), 2.29-2.36 (m, 1H), 2.43-2.48 (m, 1H), 3.03 (s, 6H), 3.84-3.88 (m, 1H), 4.10 (q, J=7.3 Hz, 1H), 4.59 (bs, 2H), 6.08 (dd, J=7.3, 4.0 Hz, 1H), 7.18 (t, J=7.3 Hz, 1H), 7.26 (t, J=7.6 Hz, 2H), 7.35 (d, J=7.6 Hz, 2H), 7.82 (s, 1H), 7.97 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.7, 30.5, 2×36.9, 43.0, 68.5, 83.8, 113.1, 126.4, 2×127.4, 2×128.0, 136.1, 140.8, 150.6, 153.9, 159.0.

Example 24 2-(Dimethylamino)-6-(3-hydroxybenzylamino)-9-(tetrahydrofuran-2-yl)-9H-purine

(39) Prepared from 2-chloro-6-(3-hydroxybenzylamino)-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.18H.sub.22N.sub.6O.sub.2, yield (%): 53. HPLC-UV/VIS retention time, purity (min., %): 25.73, 98.0. ESI.sup.+-MS m/z (rel. int. %, ion): 354.8 (100, [M+H].sup.+), 284.8 (53, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-1.99 (m, 1H), 2.21-2.27 (m, 1H), 2.30-2.37 (m, 1H), 2.43-2.47 (m, 1H), 3.03 (s, 6H), 3.86 (q, J=6.9 Hz, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.52 (bs, 2H), 6.09 (dd, J=7.0, 4.0 Hz, 1H), 6.56 (d, J=8.3 Hz, 1H), 6.75 (s, 1H), 6.76 (s, 1H), 7.05 (t, J=7.8 Hz, 1H), 7.82 (s, 1H), 7.89 (bs, 1H), 9.24 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.8, 30.5, 2×36.9, 42.9, 68.5, 83.8, 113.1, 113.4, 114.1, 118.02, 129.0, 136.1, 142.2, 150.6, 153.9, 157.2, 159.0.

Example 25 2-(Dimethylamino)-6-[(4-hydroxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(40) Prepared from 2-chloro-6-[(4-hydroxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.18H.sub.22N.sub.6O.sub.2, yield (%): 77. HPLC-UV/VIS retention time, purity (min., %): 25.50, 99.07. ESI.sup.+-MS m/z (rel. int. %, ion): 354.8 (100, [M+H].sup.+), 284.8 (54, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-2.00 (m, 1H), 2.21-2.26 (m, 1H), 2.29-2.37 (m, 1H), 2.44-2.48 (m, 1H), 3.06 (s, 6H), 3.86 (td, J=7.6, 5.8 Hz, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.49 (bs, 2H), 6.08 (dd, J=7.2, 3.8 Hz, 1H), 6.65 (d, J=8.6 Hz, 2H), 7.16 (d, J=8.3 Hz, 2H), 7.73 (bs, 1H), 7.79 (s, 1H), 9.16 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.6, 30.5, 2×36.9, 42.4, 68.4, 83.7, 113.0, 2×114.7, 2×128.6, 130.9, 135.9, 150.5, 153.8, 155.9, 159.0.

Example 26 2-(Methylamino)-6-[(2-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(41) Prepared from 2-chloro-6-[(2-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.18H.sub.22N.sub.6O.sub.2, yield (%): 77. HPLC-UV/VIS retention time, purity (min., %): 24.62, 99.0. ESI.sup.+-MS m/z (rel. int. %, ion): 355.8 (100, [M+H].sup.+), 285.8 (20, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.95-2.00 (m, 1H), 2.22-2.26 (m, 1H), 2.31-2.37 (m, 1H), 2.44 (bs, 1H), 2.70 (d, J=4.3 Hz, 3H), 3.81 (s, 3H), 3.84-3.88 (m, 1H), 4.11 (q, J=7.1 Hz, 1H), 4.59 (bs, 2H), 6.08 (dd, J=6.9, 3.8 Hz, 1H), 6.29 (d, J=3.7 Hz, 1H), 6.83 (t, J=7.3 Hz, 1H), 6.95 (d, J=8.3 Hz, 1H), 7.13 (d, J=7.3 Hz, 1H), 7.16-7.19 (m, 1H), 7.49 (bs, 1H), 7.81 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.6, 28.1, 30.7, 37.8, 55.2, 68.4, 83.6, 110.1, 113.6, 119.9, 126.9, 127.5, 127.8, 135.5, 150.5, 154.7, 156.5, 159.8.

Example 27 2-(Dimethylamino)-6-[(2-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(42) Prepared from 2-chloro-6-[(2-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.2, yield (%): 80. HPLC-UV/VIS retention time, purity (min., %): 28.95, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 369.8 (100, [M+H].sup.+), 299.8 (63, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-2.01 (m, 1H), 2.21-2.28 (m, 1H), 2.30-2.37 (m, 1H), 2.44-2.47 (m, 1H), 3.00 (s, 6H), 3.81 (s, 3H), 3.84-3.88 (m, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.61 (bs, 2H), 6.09 (dd, J=7.3, 4.0 Hz, 1H), 6.83 (t, J=7.3 Hz, 1H), 6.95 (d, J=7.9 Hz, 1H), 7.16-7.19 (m, 2H), 7.60 (bs, 1H), 7.82 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.7, 30.5, 2×36.8, 37.7, 55.2, 68.5, 83.8, 110.2, 113.1, 119.9, 2×127.5, 127.9, 136.1, 150.6, 154.1, 156.5, 159.0.

Example 28 2-(Methylamino)-6-[(4-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(43) Prepared from 2-chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.18H.sub.22N.sub.6O.sub.2, yield (%): 73. HPLC-UV/VIS retention time, purity (min., %): 23.85, 98.4. ESI.sup.+-MS m/z (rel. int. %, ion): 355.8 (100, [M+H].sup.+), 285.8 (20, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.93-2.01 (m, 1H), 2.18-2.25 (m, 1H), 2.29-2.36 (m, 1H), 2.41 (bs, 1H), 2.75 (d, J=4.6 Hz, 3H), 3.69 (s, 3H), 3.83-3.87 (m, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.52 (bs, 2H), 6.06 (dd, J=7.0, 4.0 Hz, 1H), 6.30 (bs, 1H), 6.83 (d, J=8.6 Hz, 2H), 7.27 (d, J=8.6 Hz, 2H), 7.74 (bs, 1H), 7.79 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.6, 28.2, 30.7, 42.0, 54.9, 68.4, 83.6, 3×113.4, 2×128.7, 132.6, 135.4, 150.6, 154.3, 157.9, 159.8.

Example 29 2-(Dimethylamino)-6-[(4-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(44) Prepared from 2-chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.2, yield (%): 81. HPLC-UV/VIS retention time, purity (min., %): 28.33, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 369.8 (100, [M+H].sup.+), 299.8 (74, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-1.99 (m, 1H), 2.20-2.26 (m, 1H), 2.29-2.36 (m, 1H), 2.43-2.47 (m, 1H), 3.05 (s, 6H), 3.68 (s, 3H), 3.85 (td, J=7.6, 5.8 Hz, 1H), 4.10 (q, J=7.2 Hz, 1H), 4.52 (bs, 2H), 6.08 (dd, J=7.2, 3.8 Hz, 1H), 6.83 (d, J=8.9 Hz, 2H), 7.28 (d, J=8.6 Hz, 2H), 7.81 (s, 1H), 7.87 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.7, 30.5, 2×36.9, 42.4, 55.1, 68.5, 83.8, 113.1, 2×113.4, 2×128.7, 132.7, 136.0, 150.5, 153.8, 157.9, 159.0.

Example 30 2-(Dimethylamino)-6-[(4-hydroxy-3-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine

(45) Prepared from 2-chloro-6-[(4-hydroxy-3-methoxybenzyl)amino]-9-(tetrahydrofuran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.3, yield (%): 68. HPLC-UV/VIS retention time, purity (min., %): 25.52, 99.7. ESI.sup.+-MS m/z (rel. int. %, ion): 384.8 (100, [M+H].sup.+), 314.8 (46, [M-THF+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.94-2.01 (m, 1H), 2.19-2.27 (m, 1H), 2.30-2.37 (m, 1H), 2.44-2.47 (m, 1H), 3.07 (s, 6H), 3.71 (s, 3H), 3.86 (td, J=7.7, 5.7 Hz, 1H), 4.10 (q, J=7.3 Hz, 1H), 4.50 (bs, 2H), 6.08 (dd, J=7.0, 4.0 Hz, 1H), 6.66 (d, J=7.9 Hz, 1H), 6.76 (dd, J=7.9, 1.8 Hz, 1H), 6.99 (d, J=1.8 Hz, 1H), 7.73 (bs, 1H), 7.80 (s, 1H), 8.71 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 24.7, 30.5, 2×36.9, 42.8, 55.4, 68.4, 83.7, 112.2, 113.0, 115.0, 120.0, 131.5, 135.9, 145.1, 147.1, 150.5, 153.8, 159.0.

Example 31 2-(Methylamino)-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(46) Prepared from 2-chloro-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.16H.sub.20N.sub.6O.sub.2, yield (%): 52. HPLC-UV/VIS retention time, purity (min., %): 23.42, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 328.9 (100, [M+H].sup.+), 244.9 (28, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.60 (m, 2H), 1.63-1.72 (m, 1H), 1.84-1.87 (m, 1H), 1.92-1.94 (m, 1H), 2.12-2.20 (m, 1H), 2.78 (d, J=4.9 Hz, 3H), 3.57-3.62 (m, 1H), 3.96-3.99 (m, 1H), 4.63 (bs, 2H), 5.43 (d, J=9.5 Hz, 1H), 6.22 (d, J=3.1 Hz, 1H), 6.34 (m, 2H), 7.51 (d, J=0.9 Hz, 1H), 7.62 (bs, 1H), 7.87 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.5, 24.5, 28.2, 30.1, 36.3, 67.5, 80.2, 106.5, 110.3, 112.9, 134.9, 141.5, 150.9, 153.4, 154.2, 159.8.

Example 32 2-(Dimethylamino)-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(47) Prepared from 2-chloro-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.17H.sub.22N.sub.6O.sub.2, yield (%): 96. HPLC-UV/VIS retention time, purity (min., %): 28.07, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 343.0 (100, [M+H].sup.+), 258.9 (88, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.56 (m, 2H), 1.66-1.70 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.19 (ddd, J=23.9, 12.6, 4.1 Hz, 1H), 3.07 (s, 6H), 3.58-3.64 (m, 1H), 3.97 (dd, J=10.9, 2.0 Hz, 1H), 4.59 (bs, 2H), 5.45 (dd, J=11.0, 2.1 Hz, 1H), 6.21 (d, J=3.4 Hz, 1H), 6.34 (dd, J=3.2, 2.0 Hz, 1H), 7.51 (d, J=1.2 Hz, 1H), 7.79 (bs, 1H), 7.90 (s, 1H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.1, 36.4, 2×36.9, 67.5, 80.2, 106.4, 110.3, 112.4, 135.4, 141.5, 150.9, 153.5, 153.7, 159.1.

Example 33 2-(Methylamino)-6-[(5-methylfurfuryl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(48) Prepared from 2-chloro-6-[(5-methylfurfuryl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. Pale yellow solid, chemical formula: C.sub.17H.sub.22N.sub.6O.sub.2, yield (%): 79. HPLC-UV/VIS retention time, purity (min., %): 24.98, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 343.8 (100, [M+H].sup.+), 259.7 (32, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.59 (m, 2H), 1.65-1.69 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.12-2.18 (m, 1H), 2.19 (s, 3H), 2.77 (d, J=4.6 Hz, 3H), 3.57-3.62 (m, 1H), 3.97 (dd, J=10.9, 2.0 Hz, 1H), 4.54 (bs, 2H), 5.42 (d, J=10.1 Hz, 1H), 5.93 (d, J=1.8 Hz, 1H), 6.08 (d, J=2.8 Hz, 1H), 6.37 (bs, 1H), 7.61 (vbs, 1H), 7.88 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 13.3, 22.6, 24.6, 28.2, 30.2, 36.2, 67.6, 80.1, 106.3, 107.4, 112.9, 134.9, 150.1, 150.8, 151.5, 154.1, 159.8.

Example 34 2-(Dimethylamino)-6-[(5-methylfurfuryl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(49) Prepared from 2-chloro-6-[(5-methylfurfuryl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. Pale yellow solid, chemical formula: C.sub.18H.sub.24N.sub.6O.sub.2, yield (%): 86. HPLC-UV/VIS retention time, purity (min., %): 28.90, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 357.8 (100, [M+H].sup.+), 273.7 (20, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.56 (m, 2H), 1.64-1.72 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.15-2.18 (m, 1H), 2.19 (s, 3H), 3.08 (s, 6H), 3.58-3.63 (m, 1H), 3.97 (dd, J=11.0, 1.8 Hz, 1H), 4.53 (bs, 2H), 5.45 (dd, J=11.0, 2.1 Hz, 1H), 5.93 (dd, J=2.9, 0.8 Hz, 1H), 6.08 (d, J=2.8 Hz, 1H), 7.71 (bs, 1H), 7.90 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 13.2, 22.6, 24.6, 30.1, 36.3, 2×36.9, 67.5, 80.2, 106.3, 107.3, 112.4, 135.4, 150.0, 150.7, 151.6, 153.6, 159.1.

Example 35 2-(Dimethylamino)-6-benzylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(50) Prepared from 2-chloro-6-[(3-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O, yield (%): 86. HPLC-UV/VIS retention time, purity (min., %): 29.12, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 353.8 (100, [M+H].sup.+), 269.7 (78, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.51-1.57 (m, 2H), 1.63-1.72 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.19 (ddd, J=24.0, 12.5, 3.7 Hz, 1H), 3.04 (s, 6H), 3.58-3.63 (m, 1H), 3.97 (dd, J=11.0, 1.8 Hz, 1H), 4.60 (bs, 2H), 5.45 (dd, J=11.0, 2.1 Hz, 1H), 7.18 (t, J=7.3 Hz, 1H), 7.27 (t, J=7.5 Hz, 2H), 7.35 (d, J=7.3 Hz, 2H), 7.89 (s, 1H), 7.96 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.1, 2×36.9, 43.0, 67.5, 80.2, 112.4, 126.4, 2×127.4, 2×128.0, 135.3, 140.8, 150.6, 153.9, 159.2.

Example 36 2-(Dimethylamino)-6-[(4-methylbenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(51) Prepared from 2-chloro-6-[(4-methylbenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.20H.sub.26N.sub.6O, yield (%): 82. HPLC-UV/VIS retention time, purity (min., %): 30.32, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 367.8 (100, [M+H].sup.+), 283.8 (29, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.51-1.59 (m, 2H), 1.63-1.72 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.14-2.19 (m, 1H), 2.23 (s, 3H), 3.04 (s, 6H), 3.58-3.63 (m, 1H), 3.97 (dd, J=11.0, 1.8 Hz, 1H), 4.55 (bs, 2H), 5.44 (dd, J=11.0, 2.1 Hz, 1H), 7.07 (d, J=7.9 Hz, 2H), 7.23 (d, J=7.6 Hz, 2H), 7.89 (s, 2H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 20.6, 22.6, 24.6, 30.1, 2×36.9, 42.7, 67.5, 80.2, 112.4, 2×127.4, 2×128.6, 135.3, 135.4, 137.7, 150.6, 153.8, 159.2.

Example 37 2-(Dimethylamino)-6-{[4-(trifluoromethyl)benzyl]amino}-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(52) Prepared from 2-chloro-6-{[4-(trifluoromethyl)benzyl]amino}-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.20H.sub.23F.sub.3N.sub.6O, yield (%): 96. HPLC-UV/VIS retention time, purity (min., %): 30.43, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 421.7 (100, [M+H].sup.+), 337.8 (81, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.58 (m, 2H), 1.64-1.70 (m, 1H), 1.85 (dd, J=12.8, 1.8 Hz, 1H), 1.93 (bd, J=13.4 Hz, 1H), 2.15-2.23 (m, 1H), 3.00 (s, 6H), 3.58-3.63 (m, 1H), 3.97 (d, J=11.0 Hz, 1H), 4.66 (bs, 2H), 5.45 (dd, J=10.9, 1.7 Hz, 1H), 7.54 (d, J=8.3 Hz, 2H), 7.64 (d, J=7.9 Hz, 2H), 7.91 (s, 1H), 8.10 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.1, 2×36.8, 42.8, 67.5, 80.3, 112.4, 124.4 (q, .sup.1J.sub.CF=272.1 Hz), 2×125.0 (q, .sup.3J.sub.CF=3.6 Hz), 127.1 (q, .sup.2J.sub.CF=32.2 Hz), 2×127.9, 135.5, 145.7, 150.7, 153.8, 159.1.

Example 38 2-(Dimethylamino)-6-[(3-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(53) Prepared from 2-chloro-6-[(3-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.2, yield (%): 94. HPLC-UV/VIS retention time, purity (min., %): 26.50, 99.5. ESI.sup.+-MS m/z (rel. int. %, ion): 368.8 (100, [M+H].sup.+), 284.8 (53, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.58 (m, 2H), 1.63-1.73 (m, 1H), 1.83-1.86 (m, 1H), 1.90-1.95 (m, 1H), 2.15-2.23 (m, 1H), 3.04 (s, 6H), 3.58-3.63 (m, 1H), 3.96-3.98 (m, 1H), 4.53 (bs, 2H), 5.45 (dd, J=11.0, 1.8 Hz, 1H), 6.55-6.57 (m, 1H), 6.75 (s, 1H), 6.76 (bs, 1H), 7.05 (t, J=7.9 Hz, 1H), 7.89 (s, 2H), 9.24 (bs, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.2, 36.9, 38.9, 39.1, 39.3, 39.5, 39.6, 39.8, 40.0, 42.9, 67.5, 80.3, 112.4, 113.4, 114.1, 118.0, 129.0, 135.3, 142.2, 150.7, 153.9, 157.2, 159.2.

Example 39 2-(Dimethylamino)-6-[(4-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(54) Prepared from 2-chloro-6-[(4-hydroxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. Pale yellow solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.2, yield (%): 76. HPLC-UV/VIS retention time, purity (min., %): 26.12, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 369.8 (100, [M+H].sup.+), 285.7 (83, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.51-1.56 (m, 2H), 1.66-1.70 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.18 (ddd, J=24.0, 12.5, 3.7 Hz, 1H), 3.06 (s, 6H), 3.58-3.63 (m, 1H), 3.95-3.98 (m, 1H), 4.48 (bs, 2H), 5.44 (dd, J=11.0, 1.5 Hz, 1H), 6.65 (d, J=8.6 Hz, 2H), 7.16 (d, J=8.3 Hz, 2H), 7.80 (bs, 1H), 7.88 (s, 1H), 9.19 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.1, 2×36.9, 42.4, 67.5, 80.2, 112.4, 2×114.8, 2×128.7, 130.9, 135.2, 150.6, 153.8, 156.0, 159.2.

Example 40 2-(Methylamino)-6-[(4-methoxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(55) Prepared from 2-chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 8M solution of methylamine in absolute ethanol. White solid, chemical formula: C.sub.19H.sub.24N.sub.6O.sub.2, yield (%): 70. HPLC-UV/VIS retention time, purity (min., %): 24.92, 98.0. ESI.sup.+-MS m/z (rel. int. %, ion): 369.8 (100, [M+H].sup.+), 285.8 (29, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.51-1.57 (m, 2H), 1.62-1.72 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.16 (ddd, J=24.1, 12.4, 3.5 Hz, 1H), 2.76 (d, J=4.9 Hz, 3H), 3.56-3.62 (m, 1H), 3.69 (s, 3H), 3.97 (dd, J=11.0, 1.8 Hz, 1H), 4.53 (bs, 2H), 5.41 (d, J=10.4 Hz, 1H), 6.33 (bs, 1H), 6.83 (d, J=8.6 Hz, 2H), 7.27 (d, J=8.3 Hz, 2H), 7.76 (bs, 1H), 7.86 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 28.3, 30.2, 42.1, 54.9, 67.6, 80.1, 112.9, 2×113.4, 2×128.7, 132.6, 134.8, 150.6, 154.3, 157.9, 159.9.

Example 41 2-(Dimethylamino)-6-[(4-methoxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(56) Prepared from 2-chloro-6-[(4-methoxybenzyl)amino]-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 5.6M solution of dimethylamine in absolute ethanol. White solid, chemical formula: C.sub.20H.sub.26N.sub.6O.sub.2, yield (%): 93. HPLC-UV/VIS retention time, purity (min., %): 28.85, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 383.8 (100, [M+H].sup.+), 299.8 (82, [M-THP+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.51-1.59 (m, 2H), 1.63-1.72 (m, 1H), 1.83-1.86 (m, 1H), 1.91-1.94 (m, 1H), 2.18 (ddd, J=24.0, 12.5, 4.0 Hz, 1H), 3.06 (s, 6H), 3.58-3.63 (m, 1H), 3.68 (s, 3H), 3.97 (dd, J=11.0, 1.8 Hz, 1H), 4.52 (bs, 2H), 5.44 (dd, J=11.0, 2.1 Hz, 1H), 6.83 (d, J=8.9 Hz, 2H), 7.28 (d, J=8.6 Hz, 2H), 7.88 (s, 2H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.6, 30.1, 2×36.9, 42.4, 54.9, 67.5, 80.2, 112.4, 2×113.4, 2×128.7, 132.7, 135.2, 150.6, 153.8, 157.9, 159.2.

(57) B) Substitution with 2-(dimethylamino)ethylamine or 3-(dimethylamino)propylamine—Typical Procedure

(58) A mixture of 2-chloro-6-substituted-9-(2-oxacycloalkyl)-9H-purine (150 mg, 1 eq.) and 2-(dimethylamino)ethylamine or 3-(dimethylamino)propylamine (10 eq.) in n-butanol (2.2 ml) was heated under Ar atmosphere in ACE pressure tube at 120° C. for 18 hours. The reaction mixture was evaporated under reduced pressure and the crude material was purified by silica column chromatography using chloroform/methanol as a mobile phase starting from pure chloroform with methanol gradient.

Example 42 2-{[2-(Dimethylamino)ethyl]amino}-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

(59) Prepared from compound 2-chloro-6-furfurylamino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine and 2-(dimethylamino)ethylamine. White solid, chemical formula: C.sub.19H.sub.27N.sub.7O.sub.2, yield (%): 78. HPLC-UV/VIS retention time, purity (min., %): 16.27, 99.9. ESI.sup.+-MS m/z (rel. int. %, ion): 385.8 (100, [M+H].sup.+). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ (ppm): 1.52-1.57 (m, 2H), 1.62-1.68 (m, 1H), 1.84-1.87 (m, 1H), 1.92-1.94 (m, 1H), 2.15-2.23 (m, 7H), 2.37 (t, J=6.9 Hz, 2H), 3.30-3.35 (m, 2H), 3.56-3.61 (m, 1H), 3.96-3.99 (m, 1H), 4.62 (bs, 2H), 5.41 (dd, J=11.0, 1.8 Hz, 1H), 6.17 (bs, 1H), 6.20 (dd, J=3.1, 0.6 Hz, 1H), 6.34 (dd, J=3.4, 1.8 Hz, 1H), 7.51 (dd, J=1.7, 0.8 Hz, 1H), 7.65 (bs, 1H), 7.87 (s, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) δ (ppm): 22.6, 24.5, 30.0, 36.3, 3×45.1, 58.3, 67.5, 80.3, 106.3, 110.31, 113.0, 135.0, 141.5, 150.9, 153.4, 154.2, 159.1.

(60) TABLE-US-00001 TABLE 1 Examples of 9-(oxetan-2-yl) purine derivatives PURINE SUBSTITUENT CHN ANALYSES MS ANALYSIS Comp. C2 C6 [% C, % H, % N] [M + H].sup.+ 1 MeNH— furfurylamino 60.0, 5.4, 28.0 301 2 EtNH— furfurylamino 57.3, 5.8, 26.7 315 3 Me.sub.2N— furfurylamino 57.3, 5.8 26.7 315 4 Et.sub.2N— furfurylamino 59.6, 6.5, 24.5 343 5 Me.sub.2N-ethyl-NH— furfurylamino 57.1, 6.5, 27.4 358 6 MeNH— benzylamino 61.9, 5.9, 27.1 311 7 EtNH— benzylamino 63.0, 6.2, 25.9 325 8 Me.sub.2N— benzylamino 63.0, 6.2, 25.9 325 9 Et.sub.2N— benzylamino 64.8, 6.9, 23.9 353 10 Me.sub.2N-ethyl-NH— benzylamino 62.1, 6.9, 26.7 368 11 MeNH— (3-methylbut-2-en-1-yl)amino 58.3, 7.0, 29.2 289 12 EtNH— (3-methylbut-2-en-1-yl)amino 59.6, 7.3, 27.8 303 13 Me.sub.2N— (3-methylbut-2-en-1-yl)amino 59.6, 7.3, 27.8 303 14 Et.sub.2N— (3-methylbut-2-en-1-yl)amino 61.8, 7.9, 25.4 331 15 Me.sub.2N-ethyl-NH— (3-methylbut-2-en-1-yl)amino 59.1, 7.9, 28.4 346 16 MeNH— (E)-(4-hydroxy-3-methylbut-2- 55.3, 6.6, 27.6 305 en-1-yl)amino 17 EtNH— (E)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 18 Me.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 19 Et.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 20 Me.sub.2N-ethyl-NH— (E)-(4-hydroxy-3-methylbut-2- 56.5, 7.5, 27.1 362 en-1-yl)amino 21 MeNH— (Z)-(4-hydroxy-3-methylbut-2- 55.3, 6.6, 27.6 305 en-1-yl)amino 22 EtNH— (Z)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 23 Me.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 24 Et.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 25 Me.sub.2N-ethyl-NH— (Z)-(4-hydroxy-3-methylbut-2- 56.5, 7.5, 27.1 362 en-1-yl)amino

(61) TABLE-US-00002 TABLE 2 Examples of 9-(tetrahydrofuran-2-yl) purine derivatives PURINE SUBSTITUENT CHN ANALYSES MS ANALYSIS Comp. C2 C6 [% C, % H, % N] [M + H].sup.+ 26 MeNH— furfurylamino 57.3, 5.8, 26.7 315 27 EtNH— furfurylamino 58.2, 6.1, 25.6 329 28 propylNH— furfurylamino 59.6, 6.5, 24.5 343 29 2-propylNH— furfurylamino 59.6, 6.5, 24.5 343 30 Me.sub.2N— furfurylamino 58.2, 6.1, 25.6 329 31 Et.sub.2N— furfurylamino 60.7, 6.8, 23.6 357 32 Me.sub.2N-ethyl-NH— furfurylamino 58.2, 6.8, 26.4 372 33 Me.sub.2N-propyl-NH— furfurylamino 59.2, 7.1, 25.4 386 34 MeNH— (3-methylfurfuryl)amino 58.5, 6.1, 25.6 329 35 EtNH— (3-methylfurfuryl)amino 59.6, 6.5, 24.5 343 36 Me.sub.2N— (3-methylfurfuryl)amino 59.6, 6.5, 24.5 343 37 Et.sub.2N— (3-methylfurfuryl)amino 61.6, 7.1, 22.7 371 38 Me.sub.2N-ethyl-NH— (3-methylfurfuryl)amino 59.2, 7.1, 25.4 386 39 MeNH— (4-methylfurfuryl)amino 58.5, 6.1, 25.6 329 40 EtNH— (4-methylfurfuryl)amino 59.6, 6.5, 24.5 343 41 Me.sub.2N— (4-methylfurfuryl)amino 59.6, 6.5, 24.5 343 42 Et.sub.2N— (4-methylfurfuryl)amino 61.6, 7.1, 22.7 371 43 Me.sub.2N-ethyl-NH— (4-methylfurfuryl)amino 59.2, 7.1, 25.4 386 44 MeNH— (5-methylfurfuryl)amino 58.5, 6.1, 25.6 329 45 EtNH— (5-methylfurfuryl)amino 59.6, 6.5, 24.5 343 46 Me.sub.2N— (5-methylfurfuryl)amino 59.6, 6.5, 24.5 343 47 Et.sub.2N— (5-methylfurfuryl)amino 61.6, 7.1, 22.7 371 48 Me.sub.2N-ethyl-NH— (5-methylfurfuryl)amino 59.2, 7.1, 25.4 386 49 MeNH— (3-methoxyfurfuryl)amino 55.8, 5.9, 24.4 345 50 EtNH— (3-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 51 Me.sub.2N— (3-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 52 Et.sub.2N— (3-methoxyfurfuryl)amino 59.1, 6.8, 21.8 387 53 Me.sub.2N-ethyl-NH— (3-methoxyfurfuryl)amino 56.8, 6.8, 24.4 402 54 MeNH— (4-methoxylfurfuryl)amino 55.8, 5.9, 24.4 345 55 EtNH— (4-methoxylfurfuryl)amino 57.0, 6.2, 23.5 359 56 Me.sub.2N— (4-methoxylfurfuryl)amino 57.0, 6.2, 23.5 359 57 Et.sub.2N— (4-methoxylfurfuryl)amino 59.1, 6.8, 21.8 387 58 Me.sub.2N-ethyl-NH— (4-methoxylfurfuryl)amino 56.8, 6.8, 24.4 402 59 MeNH— (5-methoxyfurfuryl)amino 55.8, 5.9, 24.4 345 60 EtNH— (5-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 61 Me.sub.2N— (5-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 62 Et.sub.2N— (5-methoxyfurfuryl)amino 59.1, 6.8, 21.8 387 63 Me.sub.2N-ethyl-NH— (5-methoxyfurfuryl)amino 56.8, 6.8, 24.4 402 64 MeNH— benzylamino 63.0, 6.2, 25.9 325 65 EtNH— benzylamino 63.9, 6.6, 24.8 339 66 Me.sub.2N— benzylamino 63.9, 6.6, 24.8 339 67 Et.sub.2N— benzylamino 65.6, 7.2, 22.9 367 68 Me.sub.2N-ethyl-NH— benzylamino 63.0, 7.1, 25.7 382 69 MeNH— (3-hydroxybenzyl)amino 60.0, 5.9, 24.7 341 70 EtNH— (3-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 71 Me.sub.2N— (3-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 72 Et.sub.2N— (3-hydroxybenzyl)amino 62.8, 6.9, 22.0 383 73 Me.sub.2N-ethyl-NH— (3-hydroxybenzyl)amino 60.4, 6.9, 24.7 398 74 MeNH— (4-hydroxybenzyl)amino 60.0, 5.9, 24.7 341 75 EtNH— (4-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 76 Me.sub.2N— (4-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 77 Et.sub.2N— (4-hydroxybenzyl)amino 62.8, 6.9, 22.0 383 78 Me.sub.2N-ethyl-NH— (4-hydroxybenzyl)amino 60.4, 6.9, 24.7 398 79 MeNH— (3-methoxybenzyl)amino 61.0, 6.3, 23.7 355 80 EtNH— (3-methoxybenzyl)amino 61.9, 6.6, 22.8 369 81 Me.sub.2N— (3-methoxybenzyl)amino 61.9, 6.6, 22.8 369 82 Et.sub.2N— (3-methoxybenzyl)amino 63.6, 7.1, 21.2 397 83 Me.sub.2N-ethyl-NH— (3-methoxybenzyl)amino 61.3, 7.1, 23.8 412 84 MeNH— (4-methoxybenzyl)amino 61.0, 6.3, 23.7 355 85 EtNH— (4-methoxybenzyl)amino 61.9, 6.6, 22.8 369 86 Me.sub.2N— (4-methoxybenzyl)amino 61.9, 6.6, 22.8 369 87 Et.sub.2N— (4-methoxybenzyl)amino 63.6, 7.1, 21.2 397 88 Me.sub.2N-ethyl-NH— (4-methoxybenzyl)amino 61.3, 7.1, 23.8 412 89 Me.sub.2N— (4-hydroxy-3- 59.3, 6.2, 21.8 385 methoxybenzyl)amino 90 MeNH— (3-methylbut-2-en-1-yl)amino 59.6, 7.3, 27.8 303 91 EtNH— (3-methylbut-2-en-1-yl)amino 60.7, 7.7, 26.6 317 92 Me.sub.2N— (3-methylbut-2-en-1-yl)amino 60.7, 7.7, 26.6, 317 93 Et.sub.2N— (3-methylbut-2-en-1-yl)amino 62.8, 8.2, 24.4 345 94 Me.sub.2N-ethyl-NH— (3-methylbut-2-en-1-yl)amino 60.1, 8.1, 27.3 360 95 MeNH— (E)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 96 EtNH— (E)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 97 Me.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 98 Et.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 60.0, 7.8, 23.3 361 en-1-yl)amino 99 Me.sub.2N-ethyl-NH— (E)-(4-hydroxy-3-methylbut-2- 57.6, 7.8, 26.1 376 en-1-yl)amino 100 MeNH— (Z)-(4-hydroxy-3-methylbut-2- 56.6, 7.0, 26.4 319 en-1-yl)amino 101 EtNH— (Z)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 102 Me.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 103 Et.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 60.0, 7.8, 23.3 361 en-1-yl)amino 104 Me.sub.2N-ethyl-NH— (Z)-(4-hydroxy-3-methylbut-2- 57.6, 7.8, 26.1 376 en-1-yl)amino 105 MeNH— (4-hydroxy-3-methylbutyl)amino 56.2, 7.6, 26.2 321 106 EtNH— (4-hydroxy-3-methylbutyl)amino 57.5, 7.8, 25.1 335 107 Me.sub.2N— (4-hydroxy-3-methylbutyl)amino 57.5, 7.8, 25.1 335 108 Et.sub.2N— (4-hydroxy-3-methylbutyl)amino 59.6, 8.3, 23.2 363 109 Me.sub.2N-ethyl-NH— (4-hydroxy-3-methylbutyl)amino 57.3, 8.3, 26.0 378

(62) TABLE-US-00003 TABLE 3 Examples of prepared 9-(tetrahydro-2H-pyran-2-yl) purine derivatives PURINE SUBSTITUENT CHN ANALYSES MS ANALYSIS Comp. C2 C6 [% C, % H, % N] [M + H].sup.+ 110 MeNH— furfurylamino 58.5, 6.1, 25.6 329 111 EtNH— furfurylamino 59.6, 6.5, 24.5 343 112 propylNH— furfurylamino 60.7, 6.8, 23.6 357 113 2-propylNH— furfurylamino 60.7, 6.8, 23.6 357 114 Me.sub.2N— furfurylamino 59.6, 6.5, 24.5 343 115 Et.sub.2N— furfurylamino 61.6, 7.1, 22.7 371 116 Me.sub.2N-ethyl-NH— furfurylamino 59.2, 7.1, 25.4 386 117 Me.sub.2N-propyl-NH— furfurylamino 60.1, 7.3, 24.5 400 118 MeNH— (3-methylfurfuryl)amino 59.6, 6.5, 24.5 343 119 EtNH— (3-methylfurfuryl)amino 60.7, 6.8, 23.6 357 120 Me.sub.2N— (3-methylfurfuryl)amino 60.7, 6.8, 23.6 357 121 Et.sub.2N— (3-methylfurfuryl)amino 62.5, 7.3, 21.9 385 122 Me.sub.2N-ethyl-NH— (3-methylfurfuryl)amino 60.1, 7.3, 24.5 400 123 MeNH— (4-methylfurfuryl)amino 59.6, 6.5, 24.5 343 124 EtNH— (4-methylfurfuryl)amino 60.7, 6.8, 23.6 357 125 Me.sub.2N— (4-methylfurfuryl)amino 60.7, 6.8, 23.6 357 126 Et.sub.2N— (4-methylfurfuryl)amino 62.5, 7.3, 21.9 385 127 Me.sub.2N-ethyl-NH— (4-methylfurfuryl)amino 60.1, 7.3, 24.5 400 128 MeNH— (5-methylfurfuryl)amino 59.6, 6.5, 24.5 343 129 EtNH— (5-methylfurfuryl)amino 60.7, 6.8, 23.6 357 130 Me.sub.2N— (5-methylfurfuryl)amino 60.7, 6.8, 23.6 357 131 Et.sub.2N— (5-methylfurfuryl)amino 62.5, 7.3, 21.9 385 132 Me.sub.2N-ethyl-NH— (5-methylfurfuryl)amino 60.1, 7.3, 24.5 400 133 MeNH— (3-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 134 EtNH— (3-methoxyfurfuryl)amino 58.1, 6.5, 22.6 373 135 Me.sub.2N— (3-methoxyfurfuryl)amino 58.1, 6.5, 22.6 373 136 Et.sub.2N— (3-methoxyfurfuryl)amino 60.0, 7.1, 21.0 401 137 Me.sub.2N-ethyl-NH— (3-methoxyfurfuryl)amino 57.8, 7.0, 23.6 416 138 MeNH— (4-methoxylfurfuryl)amino 57.0, 6.2, 23.5 359 139 EtNH— (4-methoxylfurfuryl)amino 58.1, 6.5, 22.6 373 140 Me.sub.2N— (4-methoxylfurfuryl)amino 58.1, 6.5, 22.6 373 141 Et.sub.2N— (4-methoxylfurfuryl)amino 60.0, 7.1, 21.0 401 142 Me.sub.2N-ethyl-NH— (4-methoxylfurfuryl)amino 57.8, 7.0, 23.6 416 143 MeNH— (5-methoxyfurfuryl)amino 57.0, 6.2, 23.5 359 144 EtNH— (5-methoxyfurfuryl)amino 58.1, 6.5, 22.6 373 145 Me.sub.2N— (5-methoxyfurfuryl)amino 58.1, 6.5, 22.6 373 146 Et.sub.2N— (5-methoxyfurfuryl)amino 60.0, 7.1, 21.0 401 147 Me.sub.2N-ethyl-NH— (5-methoxyfurfuryl)amino 57.8, 7.0, 23.6 416 148 MeNH— benzylamino 63.9, 6.6, 24.8 339 149 EtNH— benzylamino 64.8, 6.9, 23.9 353 150 Me.sub.2N— benzylamino 64.8, 6.9, 23.9 353 151 Et.sub.2N— benzylamino 66.3, 7.4, 22.1 381 152 Me.sub.2N-ethyl-NH— benzylamino 63.8, 7.4, 24.8 396 153 MeNH— (3-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 154 EtNH— (3-hydroxybenzyl)amino 61.9, 6.6, 22.8 369 155 Me.sub.2N— (3-hydroxybenzyl)amino 61.9, 6.6, 22.8 369 156 Et.sub.2N— (3-hydroxybenzyl)amino 63.6, 7.2, 21.2 397 157 Me.sub.2N-ethyl-NH— (3-hydroxybenzyl)amino 61.3, 7.1, 23.8 412 158 MeNH— (4-hydroxybenzyl)amino 61.0, 6.3, 23.7 355 159 EtNH— (4-hydroxybenzyl)amino 61.9, 6.6, 22.8 369 160 Me.sub.2N— (4-hydroxybenzyl)amino 61.9, 6.6, 22.8 369 161 Et.sub.2N— (4-hydroxybenzyl)amino 63.6, 7.2, 21.2 397 162 Me.sub.2N-ethyl-NH— (4-hydroxybenzyl)amino 61.3, 7.1, 23.8 412 163 MeNH— (3-methoxybenzyl)amino 61.9, 6.6, 22.8 369 164 EtNH— (3-methoxybenzyl)amino 62.8, 6.9, 22.0 383 165 Me.sub.2N— (3-methoxybenzyl)amino 62.8, 6.9, 22.0 383 166 Et.sub.2N— (3-methoxybenzyl)amino 64.4, 7.4, 20.5 411 167 Me.sub.2N-ethyl-NH— (3-methoxybenzyl)amino 62.1, 7.3, 23.0 425 168 MeNH— (4-methoxybenzyl)amino 61.9, 6.6, 22.8 369 169 EtNH— (4-methoxybenzyl)amino 62.8, 6.9, 22.0 383 170 Me.sub.2N— (4-methoxybenzyl)amino 62.8, 6.9, 22.0 383 171 Et.sub.2N— (4-methoxybenzyl)amino 64.4, 7.4, 20.5 411 172 Me.sub.2N-ethyl-NH— (4-methoxybenzyl)amino 62.1, 7.3, 23.0 425 173 MeNH— (3-methylbut-2-en-1-yl)amino 60.7, 7.7, 26.6 317 174 EtNH— (3-methylbut-2-en-1-yl)amino 61.8, 7.9, 25.4 331 175 Me.sub.2N— (3-methylbut-2-en-1-yl)amino 61.8, 7.9, 25.4 331 176 Et.sub.2N— (3-methylbut-2-en-1-yl)amino 63.7, 8.4, 23.4 359 177 Me.sub.2N-ethyl-NH— (3-methylbut-2-en-1-yl)amino 61.1, 8.4, 26.3 374 178 MeNH— (E)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 179 EtNH— (E)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 180 Me.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 181 Et.sub.2N— (E)-(4-hydroxy-3-methylbut-2- 60.9, 8.1, 22.4 375 en-1-yl)amino 182 Me.sub.2N-ethyl-NH— (E)-(4-hydroxy-3-methylbut-2- 58.6, 8.0, 25.2 390 en-1-yl)amino 183 MeNH— (Z)-(4-hydroxy-3-methylbut-2- 57.8, 7.3, 25.3 333 en-1-yl)amino 184 EtNH— (Z)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 185 Me.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 58.9, 7.6, 24.3 347 en-1-yl)amino 186 Et.sub.2N— (Z)-(4-hydroxy-3-methylbut-2- 60.9, 8.1, 22.4 375 en-1-yl)amino 187 Me.sub.2N-ethyl-NH— (Z)-(4-hydroxy-3-methylbut-2- 58.6, 8.0, 25.2 390 en-1-yl)amino 188 MeNH— (4-hydroxy-3-methylbutyl)amino 57.5, 7.8, 25.1 335 189 EtNH— (4-hydroxy-3-methylbutyl)amino 58.6, 8.1, 24.1 349 190 Me.sub.2N— (4-hydroxy-3-methylbutyl)amino 58.6, 8.1, 24.1 349 191 Et.sub.2N— (4-hydroxy-3-methylbutyl)amino 60.6, 8.6, 22.3 377 192 Me.sub.2N-ethyl-NH— (4-hydroxy-3-methylbutyl)amino 58.3, 8.5, 25.0 392

(63) TABLE-US-00004 TABLE 4 Examples of prepared 9-(oxepan-2-yl) purine derivatives PURINE SUBSTITUENT CHN ANALYSES Comp. C2 C6 [% C, % H, % N] MS ANALYSIS 193 MeNH— furfurylamino 59.6, 6.5, 24.5 343 194 EtNH— furfurylamino 60.7, 6.8, 23.6 357 195 Me.sub.2N— furfurylamino 60.7, 6.8, 23.6 357 196 Et.sub.2N— furfurylamino 62.5, 7.3, 21.9 385 197 Me.sub.2N-ethyl-NH— furfurylamino 60.1, 7.3, 24.5 400 198 MeNH— benzylamino 64.8, 6.9, 23.9 353 199 EtNH— benzylamino 65.6, 7.2, 22.9 367 200 Me.sub.2N— benzylamino 65.6, 7.2, 22.9 367 201 Et.sub.2N— benzylamino 67.0, 7.7, 21.3 395 202 Me.sub.2N-ethyl-NH— benzylamino 64.5, 7.6, 23.9 410

Example 43 In Vitro Cytotoxic Activity of Novel Compounds

(64) Low cytotoxicity of the compounds is the major property determining the agricultural use. One of the parameters used, as the basis for cytotoxicity assays, is the metabolic activity of viable cells. For example, a microtiter assay, which uses the Calcein AM, is now widely used to quantitate cell proliferation and cytotoxicity. For instance, this assay is used in drug screening programs and in chemosensitivity testing. Because only metabolically active cells cleave Calcein AM, these assays detect viable cells exclusively. The quantity of reduced Calcein AM corresponds to the number of vital cells in the culture.

(65) Mouse fibroblasts NIH3T3; mouse immortalized bone marrow macrophages B2.4 and B10A.4, and BJ (human foreskin fibroblasts) were used for routine screening of compounds. The cells were maintained in Nunc/Corning 80 cm.sup.2 plastic tissue culture flasks and cultured in cell culture medium (DMEM with 5 g/l glucose, 2 mM glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 10% fetal calf serum and sodium bicarbonate).

(66) The cell suspensions that were prepared and diluted according to the particular cell type and the expected target cell density (2.500-30.000 cells per well based on cell growth characteristics) were added by pipette (80 μl) into 96/well microtiter plates. Inoculates were allowed a pre-incubation period of 24 hours at 37° C. and 5% CO.sub.2 for stabilisation. Four-fold dilutions of the intended test concentration were added at time zero in 20 μl aliquots to the microtiter plate wells. Usually, the compound tested was evaluated at six 4-fold dilutions. In routine testing, the highest well concentration was 166.7 μM, but it can be the matter of change dependent on the agent. All compound concentrations were examined in duplicates. Incubations of cells with the tested compounds lasted for 72 hours at 37° C., in 5% CO.sub.2 atmosphere and 100% humidity. At the end of the incubation period, the cells were assayed by using Calcein AM. Ten microliters of the stock solution were pipetted into each well and incubated for 1 hour. Fluorescence (FD) was measured with the Labsystem FIA Reader Fluoroscan Ascent (UK). The tumour cell survival (GI.sub.50) was calculated using the following equitation: TCS=(FD.sub.drug exposed well/mean FD.sub.control wells)×100%. The IC.sub.50 value, the compound concentration lethal to 50% of the cells, was calculated from the obtained dose response curves (Table 5).

(67) Zero cytotoxicity is a basic prerequisite for the use of these substances in cosmetical applications. To assess the antitumor activity, the toxicity of new derivatives on panels containing cell lines of different histogenic and species origin was tested (Table 5). It has been shown that for all tested tumor lines the action of the new compounds was comparable, whereas non-malignant cell lines, NIH3T3 fibroblasts and normal human lymphocytes were resistant to this effect. The compounds listed in Table 5 can be divided into 2 groups. The first group contains “classical cytokinins” represented by 6-substituted purines (their effects are already known). The second group includes novel substituted purine derivatives. These results suggest that substitution at the 2-position of the purine skeleton generally results in a decrease in cytotoxic activity compared to “classical cytokinin” analogues. As shown in Table 5, GI50 for NIH3T3 fibroblasts and normal human lymphocytes was always greater than 166.7 μM. The new derivatives show zero toxicity for both normal and tumor cells at concentrations of about 166.7 μM and are therefore much more suitable for agricultural and cosmetic applications than “classical cytokinins” (6-substituted purine derivatives).

(68) Low cytotoxicity (high IC.sub.50 value) of the compounds is the basic prerequisite for cosmetic and medical applications. Zero cytotoxic activity was found for N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purin-2,6-diamines in comparison to classical cytokinins known in prior art (kinetin, isopentenyladenine, . . . ).

(69) TABLE-US-00005 TABLE 5 Cytotoxicity of novel compounds for different normal cell lines tested/IC.sub.50 (μmol/L) Compound B10A.4 B10A.4 BJ NIH-3T3 Kinetin >166.7 164.1 147.5 132.8 isopentenyladenine >166.7 146.9 134.1 123.7 6-benzyladenine >166.7 138.9 128.7 112.6 trans-zeatin >166.7 >166.7 >166.7 154.9 meta-topolin >166.7 178.4 165.7 167.2 ortho-topolin 87.5 94.3 107.8 94.1 Adenine >166.7 >166.7 >166.7 >166.7 3 >166.7 >166.7 >166.7 9 >166.7 >166.7 >166.7 13 >166.7 >166.7 >166.7 >166.7 14 >166.7 >166.7 >166.7 >166.7 30 >166.7 >166.7 >166.7 39 >166.7 >166.7 >166.7 46 >166.7 >166.7 >166.7 47 >166.7 >166.7 >166.7 49 >166.7 >166.7 >166.7 >166.7 58 >166.7 >166.7 >166.7 59 >166.7 >166.7 >166.7 60 >166.7 >166.7 >166.7 75 >166.7 >166.7 >166.7 82 >166.7 >166.7 >166.7 84 >166.7 >166.7 >166.7 85 >166.7 >166.7 >166.7 87 >166.7 >166.7 >166.7 93 >166.7 >166.7 >166.7 99 >166.7 >166.7 >166.7 101 >166.7 >166.7 >166.7 103 >166.7 >166.7 >166.7 104 >166.7 >166.7 >166.7 105 >166.7 >166.7 >166.7 107 >166.7 >166.7 >166.7 108 >166.7 >166.7 >166.7 111 >166.7 >166.7 >166.7 114 >166.7 >166.7 >166.7 116 >166.7 >166.7 >166.7 119 >166.7 >166.7 >166.7 127 >166.7 >166.7 >166.7 132 >166.7 >166.7 >166.7

Example 44 Radical Scavenging Activity Determined by ORAC

(70) The ability of compounds to scavenge free radicals in vitro was determined by Oxygen Radical Absorbance Capacity (ORAC) method. In brief, fluorescein (100 μl, 500 mM) and 25 μl of compound solution were added into each working well in a 96-well microplate preincubated at 37° C. Thereafter, 25 μL of 250 mM AAPH was quickly added, microplate was shaken for 5 s and the fluorescence (Ex. 485 nm, Em. 510 nm) was read every 3 min over 90 min by using microplate reader Infinite 200 (TECAN, Switzerland). The net area under the curve was used to express antioxidant activity relative to trolox which was used as a standard. Compounds with ORAC value higher than 1 are more effective than trolox, the hydrophilic equivalent of vitamin E. Kinetin, a naturally occurring cytokinin with antioxidant properties, was assigned for comparison as a control and as a substance known in the art.

(71) TABLE-US-00006 TABLE 6 Radical scavenging activity on new derivatives ORAC Compound (compound/trolox) kinetin 0.201 ± 0.5  155  2.422 ± 0.107* 30 0.292 ± 0.006 89 4.359 ± 0.183 71 5.212 ± 0.238 116 2.475 ± 0.086 110 0.205 ± 0.032 114 0.211 ± 0.018 76 0.192 ± 0.005 160 0.728 ± 0.016 128 0.461 ± 0.003 46  0.14 ± 0.004 64  0.24 ± 0.001 *Mean ± SD (n = 3)

Example 45 Activation of Transcription Factor Nrf2

(72) The ability of compounds to activate Nrf2-dependent expression was determined by EpRE-LUX reporter cell line. In brief, compounds at 100, 10, 1 and 0.1 μM concentrations were incubated for 24 h with cells. After cells lysis (10 mM Tris, 2 mM DTT), a buffer containing 0.2 mM luciferin was added to start luminescent reaction. The increase in luminescence was measured with microplate reader Infinite M200 (TECAN). Compounds with Nrf2 value higher than 1 are more effective than dimethylfumarate (DMF), a strong Nrf2 activator approved for the treatment of psoriasis and multiple sclerosis. All newly developed derivatives are more active than kinetin, a naturally-occurring cytokinin that has been used as a reference substance and is known in the art.

(73) TABLE-US-00007 TABLE 7 Activation of transcription factor Nrf2 Nrf2 Compound (compound/DMF) kinetin 0.04 ± 0.00 155  0.24 ± 0.07* 30 1.09 ± 0.29 76 0.27 ± 0.15 89 0.19 ± 0.02 71 0.18 ± 0   116 0.24 ± 0.05 110 1.83 ± 0.08 114 2.53 ± 0.39 150 0.34 ± 0.02 160 0.14 ± 0.03 128 0.66 ± 0.14 66 0.25 ± 0.17 64 1.21 ± 0.09 *Mean ± SD (n = 3)

Example 46 Inhibition of Intracellular ROS Production by Compound 114

(74) ROS production in living cells was determined by using the fluorescent probe DCFH-DA. In brief, compound 114 at 100, 10, 1 and 0.1 μM concentrations was incubated for 24 h with BJ skin fibroblasts. Thereafter, cells were washed with phosphate buffer and DCFH-DA was added. The fluorescence was read every 1 min over 20 min by using microplate reader Infinite M200 (TECAN, Switzerland). ROS production was calculated as the increase in fluorescence over 20 min. Compound 114 significantly decreased intracellular ROS production in vitro at concentration ≥10 μM.

Example 47 Protection of Peroxidation of Membrane Lipids

(75) A typical symptom associated with aging is the direct consequence of increased concentrations of reactive oxygen species and lipid peroxidation. Therefore, the malondialdehyde (MDA) levels of lipid peroxidation decomposition product were measured in separate wheat leaves which were exposed to the new prepared derivatives and kinetin for four days in the dark as described in Example 7. MDA levels were measured using thiobarbituric acid (TBA method). In detail, 100 mg of fresh plant material is homogenized with 1 ml of 80% methanol in a ball mill (MM301, Retsch, Germany) using a high shaking speed. The crude extract was centrifuged at 10,000×g for 5 minutes and 100 μl aliquots of the supernatant were mixed with 100 μl of 0.5% (w/v) TBA containing 0.1% (w/v) trichloroacetic acid. The resulting solution was then incubated for 30 min at 95° C. The samples were rapidly cooled on ice and centrifuged for 5 minutes at 1000.times.g. The supernatant absorbance was measured at 532 nm with a background reading at 600 nm and the amount of MDA-TBA complex was calculated using an absorbance coefficient (155 mM-1 cm-1). The new derivatives significantly reduced the peroxidation of membrane lipids compared to untreated control (Table 8—values expressing MDA content, membrane lipid degradation product). The compounds of the present invention significantly reduce the peroxidation level of membrane lipids in animal cells, as can be seen from the results in Table 8. The newly prepared derivatives of the general formula thus have a protective function against the negative effects of reactive oxygen species that accumulate strongly in the tissues.

(76) TABLE-US-00008 TABLE 8 Influence of new compounds on peroxidation of membrane lipids in NIH3T3 fibroblasts. Compound No. MDA (nmol/g FW) Control 18.9 (±2.2) kinetin 14.2 (±1.6) 3 12.4 (±1.1) 9 12.2 (±1.0) 13 13.1 (±1.1) 14 11.4 (±1.0) 30 12.6 (±1.1) 39 11.4 (±1.0) 46 11.2 (±1.5) 47 12.5 (±1.2) 49 12.3 (±1.0) 58 11.4 (±0.9) 59 11.5 (±1.0) 60 12.2 (±1.2) 75 11.7 (±1.2) 82 10.9 (±0.7) 84 10.6 (±0.8) 85 11.1 (±0.9) 87 10.4 (±1.0) 93 10.5 (±1.1) 99 10.2 (±0.9) 101 10.6 (±0.5) 103 10.1 (±0.9) 104 9.6 (±0.7) 105 10.6 (±0.8) 107 10.3 (±0.9) 108 9.7 (±0.6) 111 10.3 (±1.0) 114 8.8 (±0.5) 116 9.2 (±0.8) 119 9.7 (±0.9) 127 10.4 (±1.4) 132 10.7 (±1.2) *FW—fresh weight

Example 48 Ames Test

(77) The test substance was (114) assayed for the mutagenicity by the Bacterial Reverse Mutation Test. The performed test was based on EU method B.13/14 Mutagenicity—Reverse mutation test using bacteria, which is analogous to the OECD Test Guideline No. 471. Four indicator Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and one indicator Escherichia coli WP2 uvrA strain were used. The test substance was dissolved in dimethylsulfoxide (DMSO) and assayed in doses of 10-1000 μg per plate, which were applied to plates in volume of 0.1 mL. Experiments were performed without as well as with metabolic activation with a supernatant of rat liver and a mixture of cofactors. The working procedure described is in accordance with the documents Method B.13/14, Mutagenicity: Reverse Mutation Test Using Bacteria, Council Regulation (EC) No. 440/2008. Published in O.J. L 142, 2008 and OECD Test Guideline 471, Bacterial Reverse Mutation Test. Adopted Jul. 21, 1997. In the arrangement given above, the test substance was non-mutagenic for all the used tester strains without as well as with metabolic activation.

Example 49 Acute Toxicity—Fixed Dose Procedure (Subcutaneous)

(78) The aim of the study was to investigate acute toxic effects of the test substance, after a single subcutaneous administration to Wistar rats. The testing was performed according to the methods: ČSN EN ISO 10993-11: Biologické hodnoceni zdravotnických prostředků—část 11: Zkoušky na systémovou toxicitu and OECD Test Guideline No. 420 Acute Oral Toxicity—Fixed Dose Proceduře. Adopted 17 Dec. 2001. The test substance 114 was administered in a single dose as solution in vehicle (olive oil), given subcutaneously to male and female Wistar rats. The dosing was performed sequentially. The pre-test (the sighting study) was performed with 1 female and 1 male for each dose level. The dose level of 30 mg/kg was used as the starting dose. No death of animal was recorded at the starting dose, therefore the following dose level was used (100 mg/kg). The test substance administered at the dose of 30 and 100 mg/kg caused no death and no clinical signs of intoxication were observed. The presence of subcutaneously administered test substance at the dose level of 30 and 100 mg/kg was visible to second day after application. No macroscopic changes were diagnosed during pathological examination. According to the study results the Maximum Tolerated Dose (subcutaneous) of the test substance for male and female rats is >100 mg/kg.

Example 50 Acute Dermal Toxicity

(79) The test substance 114 was tested for acute dermal toxicity using Wistar rats. Testing was performed according to ČSN EN ISO 10993-11: Biologické hodnoceni zdravotnických prostředků—část 11: Zkoušky na systémovou toxicitu and according to OECD Test Guideline No. 402 Acute Dermal Toxicity Adopted 24 Feb. 1987. The study was performed as limit test: two groups of animals—5 males and 5 females at the dose of 1000 mg/kg of body weight. The pre-test was performed with 1 male and 1 female from each group. After the pilot experiment, the other animals of the group were dosed. The test substance in delivered form was applied on the shaved skin of the test animals for 24 hours. The test animals were observed 14 days after exposure to the test substance, afterwards they were sacrificed, and the necropsy for macroscopic examination of the organs was performed. The test substance applied at the dose of 1000 mg/kg of body weight did not cause death of animals. No clinical signs of toxicity were observed during the whole study. No macroscopic changes were diagnosed during pathological examination. The test substance toxicity was evaluated on the basis of mortality, body weight changes, and clinical signs of toxicity during the observation period and necropsy findings at the end of the study. According to the results of study, the value of Maximum Tolerated Dose (dermal) of the test substance for rats of both sexes is higher than 1000 mg/kg of body weight.

Example 51 In Vitro Skin Irritation Test

(80) Test substance was assayed for the in vitro skin irritation in human epidermal model EpiDerm™. The test was performed according to the Method B.46. In vitro skin irritation: Reconstructed human epidermis model test and Protocol for: In Vitro EpiDerm™ Skin Irritation Test For use with MatTek Corporation's Reconstructed Human Epidermal Model EPI-200-SIT (see par. 1.4, (1), (3)). Two types of extract were prepared by means of polar (water) as well as non-polar extractant (olive oil) according to directions given in ČSN EN ISO 10 993-12 (2012). After pre-incubation of tissues, 30 μl of an extract was placed directly atop to the tissue so it covered all tissue surfaces. Length of exposition was 60 minutes. Three tissues were used for the extract and for each control. The procedure was performed separately for each extract. After removal of extracts of the test substance from tissues, tissues were post incubated for 42 hours due to leave of damage reparation, followed by three hours incubation with MTT and two hours extraction period with shaking. Optical density (OD570) of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues. Under the above-described experimental design, average viability of tissues treated by test substance water extract was 99.0% and by test substance olive oil extract 103.0%, i.e. viability was >50% in both cases. The effect of the test substance 114 was negative in EpiDerm™ model (tissues were not damaged). According to the classification criteria given in chapter 4.5, the test substance is considered to have no category in regard to skin irritation.

Example 52 Animal Skin Irritation Test

(81) The test substance 114 was tested in Animal skin irritation test. Rabbits (New Zealand Albino breed) were used for the test. The test was performed according to ČSN EN ISO 10993 (Březen 2014): Biologické hodnoceni zdravotnických prostředků—část 10: Zkoušky dráždivosti a senzibilizace kůže, článek 6.3 Zkouška dráždivosti na zviřatech (Biological evaluation of medical devices—Part 10: Tests for irritation and delayed-type hypersensitivity). A dose 0.5 g of the test substance was applied to the skin of the back on each side of each rabbit. In order to demonstrate the sensitivity of the assay, positive control (0.5 g of lauryl sulfate) was included. At first the one rabbit (test animal. No. 16) was used. Two other rabbits (rabbit No. 17 and No. 18) were used in confirmatory test. At the end of the contact time (4 hours) the dressings were removed and animals were examined for signs of erythema and oedema, and the response was evaluated at 1, 24, 48 and 72 hours afterpatches removal. No changes in animal weights were observed. No clinical signs of systemic intoxication were detected during the whole study. Primary Irritation Index for positive control was 1.67 (slight). The reliability of positive control was ensured. No erythema and oedema were recorded at 1, 24, 48 and 72 hours in test sites in all three rabbits. Primary Irritation Index for test substance was defined as 0. No skin irritation was caused by 4-hour exposure of rabbits to test substance.

Example 53 Formulations

(82) 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 the N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine 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 epoxidised vegetable oils (epoxidised 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):

(83) TABLE-US-00009 Emulsifiable concentrates a) b) c) d) active ingredient mixture 5% 10% 25% 50% calcium dodecylbenzenesulfonate 6%  8%  6%  8% castor oil polyglycol ether 4% —  4%  4% (36 mol of ethylene oxide) octylphenol polyglycol ether —  2% —  2% (7-8 mol of ethylene oxide) cyclohexanone — — 10% 20% aromatic hydrocarbon 83%  82% 53% 18% mixture (C9-C12) Emulsions of any desired concentration can be obtained from such concentrates by dilution with water.

(84) TABLE-US-00010 Solutions a) b) c) d) active ingredient mixture  5% 10% 50% 90% 1-methoxy-3-(3-methoxy- — 20% 20% — propoxy)-propane polyethylene glycol (MW 400) 20% 10% — — N-methyl-2-pyrrolidone — — 30% 10% aromatic. hydrocarbon 75% 60% — — mixture 9C.sub.9-C.sub.12) The solutions are suitable for use in the form of microdrops.

(85) TABLE-US-00011 Wettable powders a) b) c) d) active ingredient mixture 5% 25%  50%  80% sodium lignosulfonate 4% — 3% — sodium lauryl sulfate 2% 3% —  4% sodium diisobutylnaphthalene- — 6% 5%  6% sulfonate octylphenol polyglycol ether — 1% 2% — (7-8 mol of ethylene oxide) highly dispersed silicic acid 1% 3% 5% 10% kaolin 87%  61%  37%  — 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.

(86) TABLE-US-00012 Suspension concentrates a) b) c) d) active ingredient mixture 3% 10%  25%  50%  ethylene glycol 5% 5% 5% 5% nonylphenol polyglycol ether — 1% 2% — (15 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 86%  78%  64%  38% 

(87) 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.

(88) Dry Capsules

(89) 5000 capsules, each of which contain 0.25 g of one of the N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative as active ingredient, are prepared as follows:

(90) Composition: Active ingredient: 1250 g; Talc: 180 g; Wheat starch: 120 g; Magnesium stearate: 80 g; Lactose 20 g.

(91) 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.

(92) Soft Capsules

(93) 5000 soft gelatine capsules, each of which contain 0.05 g of one of the N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative as active ingredient, are prepared as follows:

(94) Composition: 250 g Active ingredient+2 litres Lauroglycol

(95) 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 0.419 g of the mixture are then transferred to soft gelatine capsules by means of a capsule-filling machine.

(96) Soft Capsules

(97) 5000 soft gelatine capsules, each of which contain 0.05 g of one of the N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative as active ingredient, are prepared as follows:

(98) Composition: 250 g Active ingredient+1 litre PEG 400+1 litre Tween 80

(99) Preparation process: The powdered active ingredient is suspended in PEG 400 (polyethylene glycol of Mr in the range of from 380 to 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 0.43 g of the mixture are then transferred to soft gelatine capsules by means of a capsule-filling machine.

Example 54 Gel Formulation

(100) An ointment formulation was tested during a pilot clinical study with 4 volunteers with psoriatic skin disorders. The components are given in grams per 100 g.

(101) TABLE-US-00013 Compound Content Compound 114 1.0 g Butylhydroxytoluenum 0.2 g Butylparaben 0.2 g Diethyleneglycol monoethyl ether 10.0 g Silica colloidalis anhydrica 5.0 g Propylene glycol laurate 83.6 g

(102) The gel consistence may be additionally modified by addition of silica colloidalis anhydrica. It is again expected that the transdermal Transcutol P/Lauroglycol FCC system will increase the efficiency of compound 114. Silica colloidalis anhydrica will probably slow down the penetration of the active substance.

Example 55 Preparation Procedure of a Skin Ointment

(103) The formulation components are given in grams per 200 g:

(104) TABLE-US-00014 Compound Content Compound 114 2.0 g Butylhydroxytoluenum 0.4 g Butylparaben 0.4 g Diethyleneglycol monoethyl ether 20.0 g Glycerol dibehenate 44.0 g Propylene glycol laurate 133.2 g
Recommended Procedure

(105) Phase A: 2 grams of N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative 114 were dissolved in 20 g of Transcutol P while stirring continuously at room temperature in a separate glass or stainless-steel container. The dissolution process may be accelerated by heating the solution to a maximal temperature of 40° C.

(106) Phase B: 0.4 grams of Nipanox BHT and 0.4 g of Nipabutyl were dissolved while stirring continuously in 133.2 g of Lauroglycol FCC at a temperature of approximately 70° C. in another separate glass or stainless-steel container. The clear oily solution is heated to a temperature of approximately 80° C. and 44 g of Compritol 888 ATO are melted in it while stirring continuously. The clear oily solution is cooled down to approximately 60° C. and during continuous stirring and cooling down is mixed with phase A. The resulting whitish ointment-like substance is divided into approximately 15 gram portions and filled into prearranged plastic containers.

Example 56 Formulation of a Composition for Topical Application to the Skin

(107) A composition for topical application to the skin contains the following ingredients by weight %:

(108) TABLE-US-00015 Active ingredient: Compound 114 0.1% Oil phase: Cetyl alcohol 5.0% Glyceryl monostearate 15.0% Sorbitan monooleate 0.3% Polysorbate 80 USP 0.3% Aqueous phase: Methylcellulose 100 cps 1.0% Methyl paraben 0.25% Propyl paraben 0.15% Purified water q.s. to 100%

(109) Methyl paraben and propyl paraben were dissolved in hot water and subsequently methylcellulose was dispersed in the hot water. The mixture was chilled at 60° C. until the methylcellulose dissolved. The mixture was then heated to 72° C. and added to the oil phase which was heated to 70° C. while stirring continuously. N.sup.2,N.sup.6-disubstituted-9-(2-oxacycloalkyl)-9H-purine-2,6-diamine derivative 114 was added at a temperature of 35° C. and the resulting mixture was stirred continuously until dispersed. This composition is applied to the skin on at least a daily basis until the desired skin-ameliorating (anti-aging) effect is reached.