Substituted thienopyrrolopyrimidine ribonucleosides for therapeutic use

Abstract

A group of substituted thienopyrrolopyrimidine ribonucleosides of general formula I, in which R shows strong cytostatic and cytotoxic activities preferably against cancer cell lines of broad spectrum of diseases including tumors of various histogenetic origin.

Claims

1. Substituted thienopyrrolopyrimidine ribonucleosides of general formula I ##STR00015## wherein R is selected from the group consisting of: C1-C5 alkyl, optionally substituted by at least one substitutent selected from hydroxy, sulfanyl, amino, C1-C5 alkoxy, C1-C5 sulfanyl, C1-C5 alkylamino, and di(C1-C5 alkyl)amino; C2-C6 alkenyl, optionally substituted by at least one substitutent selected from hydroxy, sulfanyl, amino, C1-C5 alkoxy, C1-C5 sulfanyl, C1-C5 alkylamino, and di(C1-C5 alkyl)amino; C6-C12 aryl, optionally substituted by at least one substitutent selected from C1-C5 alkyl, hydroxy, sulfanyl, amino, C1-C5 alkoxy, C1-C5 sulfanyl, C1-C5 alkylamino, and di(C1-C5 alkyl)amino; C4-12 heteroaryl, further comprising at least one heteroatom selected from O and S; optionally substituted by at least one substitutent selected from C1-C5 alkyl, hydroxy, sulfanyl, amino, C1-C5 alkoxy, C1-C5 sulfanyl, C1-C5 alkylamino, and di(C1-C5 alkyl)amino; amino, C1-C5 alkylamino, di(C1-C5 alkyl)amino, C1-C5 alkoxy, and C1-C5 alkylsulfanyl; or a pharmaceutically acceptable salt thereof, or an optical isomer thereof; or a mixture of optical isomers.

2. Substituted thienopyrrolopyrimidine ribonucleosides of general formula I according to claim 1, wherein R is selected from the group consisting of: C1-C5 alkyl, phenyl, naphthyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, benzofuryl, dibenzofuryl, amino, C1-C5 alkylamino, di(C1-C5 alkyl)amino, C1-C5 alkoxy, and C1-C5 alkylsulfanyl.

3. Substituted thienopyrrolopyrimidine ribonucleosides of general formula I according to claim 1, wherein R is selected from the group consisting of: furan-2-yl, furan-3-yl, benzofuran-2-yl, methylsulfanyl, methoxy, amino, dimethylamino and methyl.

4. Thienopyrrolopyrimidine ribonucleosides of general formula I of claim 1 being selected from the following compounds: 4-(Furan-2-yl)-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-(Furan-3-yl)-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-(Benzofuran-2-yl)-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-Methyl-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-(N,N-dimethylamino)-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-Amino-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, 4-Methoxy-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine, and 4-(Methylsulfanyl)-8-((3-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine.

5. A method of inhibition of pathological cell proliferation of tumor/non-tumor origin, comprising the step of administering the compound of general formula I according to claim 1 to a subject in need thereof.

6. A method of treatment of a tumor disease, comprising the step of administering the compound of general formula I according to claim 1 to a subject in need thereof.

7. A pharmaceutical composition characterized in that it comprises at least one compound of general formula I according to claim 1 and at least one pharmaceutically acceptable carrier, filler and/or excipient.

8. A pharmaceutical composition according to claim 7 for use in the inhibition of pathological cell proliferation of tumor/non-tumor origin and/or in the treatment of tumor/non tumor disease associated with cell hyperproliferation.

9. A method of treatment of tumor/non-tumor disease associated with cell hyperproliferation, comprising the step of administering the compound of general formula I according to claim 1 to a subject in need thereof.

10. The method according to claim 6, wherein the tumor disease is selected from the group consisting of tumors of epitelial, mesenchymal and neuroectodermal origin.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) Compounds Numbering

(2) Following numbering of compounds is used:

(3) ##STR00003##

(4) R=

(5) ##STR00004##

(6) Synthesis of Compounds

(7) Key-intermediate benzoylated 4-chlorothienopyrrolopyrimidine ribonucleoside 6 was synthesised by 4-step procedure starting from 4,6-dichloropyrimidine (2), which was zincated (Mosrin, M.; Knochel, P. Chem. Eur. J. 2009, 15, 1468-1477) and coupled with 3-iodothiphene to give 4,6-dichloro-5-thiophen-3-ylpyrimidine (3). Nucleophilic substitution with one equivalent of sodium azide in THF furnished azido derivative 4, which was photochemically cyclized to desired thienopyrrolopyrimidine 5. Vorbrggen's glycosylation gave benzoylated 4-chlorothienopyrrolopyrimidine nucleoside 6 (Scheme 1).

(8) ##STR00005##

(9) Target 4-substituted nucleosides (Scheme 2) were prepared by palladium-catalyzed reactions or nucleophilic substitutions. 2-Furyl group was introduced into position 4 by Stille coupling with 2-furyltributylstannane, 3-furyl and 2-benzofuryl groups by Suzuki reaction with corresponding boronic acids, methyl derivative was synthesised by palladium-catalyzed alkylation with trimethylalluminium and dimethylaminoderivative by nucleophilic substitution with dimethylamine. All these reactions led to benzoylated derivatives, which gave target free nucleosides by deprotection using sodium methoxide in methanol. Methoxy, amino and methylsulfanyl groups were introduced by nucleophilic substitution, benzoyl groups were deprotected under reaction conditions.

(10) ##STR00006##

(11) TABLE-US-00001 TABLE 1 Synthesis of 4-substituted thienopyrrolopyrimidine nucleosides 7, 1 Protected Yield Free Yield Entry Conditions R nucleoside [%] nucleoside [%] 1 a 2-furyl 7a 67 1a 68 2 b 3-furyl 7b 82 1b 83 3 b 2-benzofuryl 7c 83 1c 86 4 c Me 7d 1d 70 5 d Me.sub.2N 7e 85 1e 88 6 e NH.sub.2 1f 78 7 f MeO 1g 65 8 g MeS 1h 90

(12) If tested compounds showed activity in in vitro cytotoxic test (Table 4), it was selective against broad spectrum of cancer cell lines of various histogenetic origin (mesenchymal or epitelial tumors) with significantly lower activity against normal human fibroblasts (BJ and MRC-5 cell lines). Active compounds showed promising therapeutic indexes (15-2500). IC.sub.50 values of compounds 1c, 1f were in micromolar range, IC.sub.50 values of compounds 1d, 1g, 1h, were sub-micromolar to nanomolar. Cytotoxic activity against cancer cells was independent on p53 gene status, same activities were found for HCT116 (p53 wild type) and for mutant line with deleted gene HCT116 (p53 /). However, number of derivatives showed lower cytotoxicity against cells overexpressing transport proteins (mdr-1 for K562-TAX line and mrp-1 for CEM-DNR).

EXAMPLES

(13) List of Abbreviations

(14) ATR Attenuated total reflectance

(15) aq. aqueous

(16) bd broad doublet

(17) bq broad quartet

(18) bs broad singlet

(19) bt broad triplet

(20) btd broad triplet of doublets

(21) Bz benzoyl

(22) C-18 C-18 reverse phase as stationary phase

(23) calcd calculated

(24) d doublet

(25) dd doublet of doublets

(26) ddd doublet of doublet of doublets

(27) DMF N,N-dimethylformamide

(28) DMSO dimethylsulfoxide

(29) dt doublet of triplets

(30) eq. equivalent

(31) ESI electrospray ionization

(32) EtOH ethanol

(33) FT Fourier transform

(34) HPFC high performance flash chromatography

(35) HR high resolution

(36) iPr isopropyl

(37) IR infrared spectroscopy

(38) m multiplet

(39) Me methyl

(40) MeCN acetonitrile

(41) MeOH methanol

(42) MeONa sodium methoxide

(43) MeSNa sodium thiomethoxide

(44) m.p. melting point

(45) MS mass spectrometry

(46) MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

(47) v wave number

(48) NMR nuclear magnetic resonance

(49) Ph phenyl

(50) q quartet

(51) r.t. room temperature

(52) s singlet

(53) SiO.sub.2 silicagel as stationary phase

(54) t triplet

(55) td triplet of doublets

(56) TMSOTf trimethylsilyl trifluoromethansulfonate

(57) TFA trifluoroacetic acid

(58) THF tetrahydrofuran

(59) (TMP).sub.2Zn bis(2,2,6,6-tetramethypiperidinyl)zinc

(60) General Experimental Part

(61) NMR spectra were recorded on a 400 MHz (.sup.1H at 400 MHz, .sup.13C at 100.6 MHz), a 500 MHz (.sup.1H at 500 MHz, .sup.13C at 125.7 MHz), or a 600 MHz (.sup.1H at 600 MHz, .sup.13C at 150.9 MHz) spectrometer. Melting points were determined on a Kofler block and are uncorrected. Germicid UV bulb, model EUV-13B was used for photocyclization reactions. Optical rotations were measured at 25 C., and [].sub.D.sup.20 values are given in 10.sup.1 deg cm.sup.2 g.sup.1. High resolution mass spectra were measured using electrospray ionization. Reverse-phase high performance flash chromatography (HPFC) was performed on KP-C18-HS columns with Biotage SP1 system. FT IR spectra were measured on Bruker Alpha spectrometer using ATR technique. The purity of all tested compounds was confirmed by HPLC analysis and was >95%.

(62) TABLE-US-00002 TABLE 3 List of Compounds in Examples Example Compound Structure Systematic name 10 1a 4-(Furan-2-yl)-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 11 1b 4-(Furan-3-yl)-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 12 1c 4-(Benzofuran-2-yl)-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 13 1d 0 4-Methyl-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 14 1e 4-(N,N-dimethylamino)-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 15 1f 4-Amino-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 16 1g 4-Methoxy-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine 17 1h 4-(Methylsulfanyl)-8-(-D-ribofuranosyl)-8H- thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine

(63) General Procedure A (Zempln Deprotection of Benzoylated Nucleosides):

(64) Protected nucleoside was dissolved in methanol (10 ml) and 1M solution of MeONa in MeOH (0.3 equiv.) was added. Reaction mixture was stirred at r.t. overnight. Solvent was evaporated under reduced pressure and crude products were purified using RP-HPFC (H.sub.2O/MeOH, 0%-100%, 2 L).

Example 1

4,6-Dichloro-5-(thiofen-3-yl)pyrimidine (3)

(65) 4,6-Dichloropyrimidine (2) was zincated according to modified literature procedure (Mosrin, M.; Knochel, Chem. Eur. J. 2009, 15, 1468-1477). 4,6-Dichloropyrimidine (2) (950 mg, 6.30 mmol) was dissolved in THF (10 ml) and added dropwise into an ice-cooled solution of (TMP).sub.2Zn.MgCl.sub.2.LiCl in THF (0.35 M, 9.0 ml, 3.15 mmol) and the reaction mixture was stirred at 0 C. for 1 hour, then let to warm to r.t. for one hour and added to a solution of 3-iodothiophene (0.74 ml, 6.7 mmol) and Pd(PPh.sub.3).sub.4 (775 mg, 0.67 mmol) in THF (3 ml), which was pre-stirred at r.t. for 20 min., and stirred at 65 for 16 hrs. Solvent was evaporated under reduced pressure and crude mixture was purified by HPFC (hexane/EtOAc 0.fwdarw.1%) to give 3 (890 mg, 58%) as a white solid. m.p. 178-180 C. IR (ATR): v=2932, 2862, 1510, 1404, 1326, 813, 774. .sup.1H NMR (600.1 MHz, CDCl.sub.3): 7.15 (dd, 1H, J.sub.4,5=4.9, J.sub.4,2=1.4, H-4-thienyl); 7.47 (dd, 1H, J.sub.2,5=3.0, J.sub.2,4=1.4, H-2-thienyl); 7.48 (dd, 1H, J.sub.5,4=4.9, J.sub.5,2=3.0, H-5-thienyl); 8.75 (s, 1H, H-2). .sup.13C NMR (150.9 MHz, CDCl.sub.3): 126.12 (CH-5-thienyl); 126.91 (CH-2-thienyl); 128.08 (CH-4-thienyl); 129.83 (C-5); 131.56 (C-3-thienyl); 156.44 (CH-2); 161.55 (C-4,6). APCI MS m/z (rel %): 231 (100) [M+H]. HR MS (APCI) for C.sub.8H.sub.5N.sub.2Cl.sub.2S [M+H]: calcd 230.95450; found 230.95456.

Example 2

4-Azido-6-chloro-5-(thiophen-3-yl)pyrimidine (4)

(66) 4,6-Dichloro-5-thiophen-3-ylpyrimidine (3) (1.1 g, 4.9 mmol) was dissolved in THF (10 ml), NaN.sub.3 (320 mg, 4.9 mmol) and LiCl (204 mg, 4.9 mmol) were added and the reaction mixture was stirred for 2 days at r.t. Solvent was evaporated and the crude material was purified by column chromatography on silica (hexane/EtOAc 6:1). Desired product 4 (1.0 g, 90%) was obtained as a yellow solid. m.p. 85 C.; IR (ATR): v=3390, 3086, 2148 (weak), 1587, 1514, 1406, 1382, 1324, 1182, 1086, 978, 898, 816, 794, 763, 633, 504. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 7.75 (dd, 1H, J.sub.4,5=5.1, J.sub.4,2=1.3, H-4-thienyl); 7.82 (dd, 1H, J.sub.5,4=5.1, J.sub.5,2=3.0, H-5-thienyl); 8.34 (dd, 1H, J.sub.2,5=3.0, J.sub.2,4=1.3, H-2-thienyl); 10.17 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 117.66 (C-5); 126.71 (CH-5-thienyl); 128.78 (CH-4-thienyl); 129.39 (C-3-thienyl); 130.22 (CH-2-thienyl); 137.98 (CH-2); 144.00, 150.95 (C-4,6).

Example 3

4-Chloro-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (5)

(67) Azide 4 (200 mg, 0.84 mmol)) was dissolved in TFA (20 ml) and stirred at r.t under irradiation by 4 W UV bulb for 24 h. UV bulb was placed inside the flask with the reaction mixture. Solvent was evaporated and the crude material was purified by HPFC (40 g silica cartridge, hexane/EtOAc, 20.fwdarw.30%) to give compound 5 (98 mg, 56%) as a white solid. m.p. 258-261 C. IR (ATR): v=3047, 2931, 2861, 2804, 2663, 1607, 1568, 1499, 1470, 1425, 1313, 1267, 1229, 1107, 1071, 917, 835, 783, 635. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 7.41 (d, 1H, J.sub.6,5=5.3, H-6); 7.50 (d, 1H, J.sub.5,6=5.3, H-5); 8.65 (s, 1H, H-2); 13.23 (bs, 1H, NH). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 111.11 (C-4a); 118.01 (CH-5); 119.59 (C-4b); 121.46 (CH-6); 142.54 (C-7a); 148.48 (C-4); 150.69 (CH-2); 156.69 (C-8a). APCI MS m/z (rel %): 209 (100) [M+H]. HR MS (APCI) for C.sub.8H.sub.5N.sub.3ClS [M+H]: calcd 209.98872; found 209.98874.

Example 4

4-Chloro-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (6)

(68) Tricyclic base 5 (150 mg; 0.7 mmol) was dissolved in MeCN (30 ml) and BSA (175 l, 0.7 mmol) was added. The reaction mixture was heated at 60 C. for 30 minutes, then, TMSOTf (316 l, 1.75 mmol) and 1-O-acetyl-2,3,5-tri-O-benzoyl--D-ribofuranose (724 mg, 1.4 mmol) were added. The mixture was heated to 60 C. for 12 hours. After cooling to r.t., the mixture was extracted with EtOAc and water, organic layer was washed with NaHCO.sub.3 and again with water, dried over MgSO.sub.4 and evaporated under reduced pressure. Crude product was purified using column chromatography (hexane/EtOAc, 15.fwdarw.35%). Protected nucleoside 6 (187 mg, 40%) was obtained as a white solid. m.p. 166-169 C. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 4.72 (dd, 1H, J.sub.gem=12.3 J.sub.5b,4=4.6, H-5b); 4.84 (dd, 1H, J.sub.gem=12.3, J.sub.5a,4=3.1, H-5a); 5.03 (td, 1H, J.sub.4,5=4.6, 3.1, J.sub.4,3=4.6, H-4); 6.11 (dd, 1H, J.sub.3,2=6.1, J.sub.3,4=4.6, H-3); 6.32 (dd, 1H, J.sub.2,3=6.1, J.sub.2,1=5.7, H-2); 6.96 (d, 1H, J.sub.1,2=5.7, H-1); 7.43 (m, 2H, H-m-Bz); 7.47 (d, 1H, J.sub.6,5=5.4, H-6); 7.48, 7.52 (2m, 22H, H-m-Bz); 7.53 (d, 1H, J.sub.5,6=5.4, H-5); 7.62, 7.66, 7.69 (3m, 31H, H-p-Bz); 7.82, 7.95, 7.99 (3m, 32H, H-o-Bz); 8.66 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 63.76 (CH.sub.2-5); 70.94 (CH-3); 72.53 (CH-2); 79.85 (CH-4); 86.54 (CH-1); 112.08 (C-4a); 117.97 (CH-5); 120.41 (C-4b); 123.39 (CH-6); 128.40, 128.76 (C-i-Bz); 128.85, 128.90, 128.99 (CH-m-Bz); 129.32 (C-i-Bz); 129.38, 129.46, 129.58 (CH-o-Bz); 133.73, 134.12 (CH-p-Bz); 140.99 (C-7a); 149.15 (C-4); 150.91 (CH-2); 155.64 (C-8a); 164.58, 164.90, 165.59 (CO-Bz). ESI MS m/z (rel %): 676 (100) [M+Na]. HR MS (ESI) for C.sub.34H.sub.24N.sub.3O.sub.7ClSNa [M+Na]: calcd 676.09157; found 676.09181.

Example 5

4-(2-Furyl)-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (7a)

(69) Protected nucleoside 6 (200 mg, 0.31 mmol), 2-furyltributylstannane (131 mg, 0.38 mmol) and PdCl.sub.2(PPh.sub.3).sub.2 (22 mg, 0.03 mmol) were dissolved in anhydrous DMF (5 ml) and heated to 100 C. for 6-8 hours. The volatiles were removed in vacuo and the residue was loaded on silica column containing 15% of KF. Column was washed with 3 liters of hexane, than with gradient of ethyl-acetate in hexane (0.fwdarw.20%). Protected nucleoside 7a (140 mg, 67%) was obtained as a white solid. m.p. 114-118 C. IR (ATR): v=2933, 2862, 1722, 1605, 1562, 1446, 1435, 1289, 1264, 1134, 1110, 1091, 1055, 1029, 708, 687. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 4.73 (dd, 1H, J.sub.gem=12.4, J.sub.5b,4=4.7, H-5b); 4.84 (dd, 1H, J.sub.gem=12.4, J.sub.5a,4=3.1, H-5a); 5.02 (td, 1H, J.sub.4,5=4.7, 3.1, J.sub.4,3=4.7, H-4); 6.13 (dd, 1H, J.sub.3,2=6.1, J.sub.3,4=4.7, H-3); 6.33 (dd, 1H, J.sub.2,3=6.1, J.sub.2,1=5.9, H-2); 6.84 (dd, 1H, J.sub.4,3=3.5, J.sub.4,5=1.7, H-4-furyl); 7.00 (d, 1H, J.sub.1,2=5.9, H-1); 7.40 (d, 1H, J.sub.6,5=5.4, H-6); 7.42 (m, 2H, H-m-Bz); 7.47-7.56 (m, 5H, H-3-furyl, H-m-Bz); 7.62, 7.65, 7.70 (3m, 31H, H-p-Bz); 7.81 (m, 2H, H-o-Bz); 7.93 (d, 1H, J.sub.5,6=5.4, H-5); 7.99-8.02 (m, 4H, H-o-Bz); 8.24 (dd, 1H, J.sub.5,4=1.7, J.sub.5,3=0.8, H-5-furyl); 8.76 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 63.84 (CH.sub.2-5); 70.92 (CH-3); 72.15 (CH-2); 79.51 (CH-4); 85.94 (CH-1); 107.30 (C-4a); 112.99 (CH-4-furyl); 113.63 (CH-3-furyl); 121.21 (CH-6); 121.61 (CH-5); 121.71 (C-4b); 128.39, 128.80 (C-i-Bz); 128.91, 128.94, 129.04 (CH-m-Bz); 129.41 (C-i-Bz); 129.48, 129.62 (CH-o-Bz); 133.76, 134.16 (CH-p-Bz); 140.83 (C-7a); 145.22 (C-4); 146.86 (CH-5-furyl); 151.21 (CH-2); 152.36 (C-2-furyl); 156.98 (C-8a); 164.60, 164.96, 165.67 (CO-Bz). ESI MS m/z (rel %): 686 (45) [M+H]; 708 (100) [M+Na]. HR MS (ESI) for C.sub.38H.sub.28N.sub.3O.sub.8S [M+H]: calcd 686.15916; found 686.15935.

Example 6

4-(3-Furyl)-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (7b)

(70) Protected nucleoside 6 (250 mg, 0.38 mmol), furan-3-boronic acid (85 mg, 0.76 mmol), K.sub.2CO.sub.3 (157 mg, 0.76 mmol) and Pd(PPh.sub.3).sub.4 (22 mg, 0.019 mmol) were dissolved in toluene and heated to 100 C. for 6 hours. Solvent was evaporated and the crude product was purified by column chromatography (hexane/EtOAc, 0.fwdarw.20%). Nucleoside 7b containing 30% of impurities (312 mg, 83%) was obtained as a yellow solid and was directly deprotected. ESI MS m/z (rel %): 686 (19) [M+H]; 708 (100) [M+Na]. HR MS (ESI) for C.sub.38H.sub.27N.sub.3O.sub.8SNa [M+Na]: calcd 708.14111; found 708.14120.

Example 7

4-(2-Benzofuryl)-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (7c)

(71) Compound 7c was prepared as described for 7b from protected nucleoside 6 (250 mg, 0.38 mmol) and benzofuran-2-boronic acid (124 mg, 0.76 mmol). Nucleoside 7c (228 mg, 82%) was obtained as a yellow solid. m.p. 110-113 C. IR (ATR): v=2965, 2938, 1728, 1603, 1454, 1433, 1290, 1267, 1112, 1070, 1030, 709, 688. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 4.75 (dd, 1H, J.sub.gem=12.4, J.sub.5b,4=4.6, H-5b); 4.86 (dd, 1H, J.sub.gem=12.4, J.sub.5a,4=3.1, H-5a); 5.04 (td, 1H, J.sub.4,5=4.6, 3.1, J.sub.4,3=4.6, H-4); 6.15 (dd, 1H, J.sub.3,2=6.1, J.sub.3,4=4.6, H-3); 6.35 (dd, 1H, J.sub.2,3=6.1, J.sub.2,1=5.8, H-2); 7.03 (d, 1H, J.sub.1,2=5.8, H-1); 7.39 (ddd, 1H, J.sub.5,4=8.0, J.sub.5,6=7.3, J.sub.5,7=1.0, H-5-benzofuryl); 7.42 (m, 2H, H-m-Bz); 7.45 (d, 1H, J.sub.6,5=5.4, H-6); 7.48-7.55 (m, 5H, H-6-benzofuryl, H-m-Bz); 7.62, 7.64, 7.70 (3m, 31H, H-p-Bz); 7.82 (m, 2H, H-o-Bz); 7.85 (ddd, 1H, J.sub.4,5=8.0, J.sub.4,6=1.3, J.sub.4,7=1.0, H-4-benzofuryl); 7.93 (d, 1H, J.sub.3,7=1.0, H-3-benzofuryl); 7.98 (dq, 1H, J.sub.7,6=8.4, J.sub.7,3=J.sub.7,4=J.sub.7,5=1.0, H-7-benzofuryl); 8.00, 8.01 (2m, 22H, H-o-Bz); 8.08 (d, 1H, J.sub.5,6=5.4, H-5); 8.86 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 63.88 (CH.sub.2-5); 70.97 (CH-3); 72.26 (CH-2); 79.61 (CH-4); 86.08 (CH-1); 108.58 (C-4a); 109.31 (CH-3-benzofuryl); 112.06 (CH-7-benzofuryl); 121.55 (C-4b); 121.68 (CH-6); 121.83 (CH-5); 122.76 (CH-4-benzofuryl); 124.21 (CH-5-benzofuryl); 126.82 (CH-6-benzofuryl); 127.83 (C-3a-benzofuryl); 128.42, 128.82 (C-i-Bz); 128.94, 128.97, 129.07 (CH-m-Bz); 129.42 (C-i-Bz); 129.50, 129.65 (CH-o-Bz); 133.79, 134.19 (CH-p-Bz); 141.60 (C-7a); 145.00 (C-4); 151.19 (CH-2); 153.99 (C-2-benzofuryl); 155.62 (C-7a-benzofuryl); 157.19 (C-8a); 164.65, 164.98, 165.70 (CO-Bz). ESI MS m/z (rel %): 758 (100) [M+Na]. HR MS (ESI) for C.sub.42H.sub.29N.sub.3O.sub.8S [M+H]: calcd 736.17481; found 736.17495.

Example 8

4-Methyl-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (7d)

(72) (Me).sub.3Al (345 l, 2M in toluene) was added to a solution of nucleoside 6 (150 mg, 0.23 mmol) and Pd(PPh.sub.3).sub.4 (12 mg, 0.012 mmol) in THF (8 ml) and the reaction mixture was stirred at 70 C. for 12 hr. Solvent was evaporated and purification by HPFC (hexane/EtOAc 10.fwdarw.50%) gave nucleoside 7d (137 mg, purity 90%), which was directly deprotected. ESI MS m/z (rel %): 634 (18) [M+H]; 656 (100) [M+Na]. HR MS (ESI) for C.sub.35H.sub.28N.sub.3O.sub.7S [M+H]: calcd 634.16425; found 634.16474.

Example 9

4-N,N-dimethylamino-8-(2,3,5-tri-O-benzoyl--D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (7e)

(73) Dimethylamine (253 l, 2M in THF) was added to a solution of nucleoside 11 (220 mg, 0.34 mmol) in isopropanol (15 ml) and the reaction mixture was stirred at r.t. for 24 hr. Solvent was evaporated and purification by HPFC (hexane/EtOAc 15%) gave nucleoside 7e (190 mg, 85%) as a white solid. m.p. 148-151 C. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.35 (s, 6H, (CH.sub.3).sub.2N); 4.70 (dd, 1H, J.sub.gem=12.3, J.sub.5b,4=4.6, H-5b); 4.81 (dd, 1H, J.sub.gem=12.3, J.sub.5a,4=3.1, H-5a); 4.96 (td, 1H, J.sub.4,5=4.6, 3.1, J.sub.4,3=4.6, H-4); 6.09 (dd, 1H, J.sub.3,2=6.1, J.sub.3,4=4.6, H-3); 6.26 (t, 1H, J.sub.2,1=J.sub.2,3=6.1, H-2); 6.90 (d, 1H, J.sub.1,2=6.1, H-1); 7.23 (d, 1H, J.sub.6,5=5.5, H-6); 7.39-7.44 (m, 3H, H-5, H-m-Bz); 7.51, 7.52 (2m, 22H, H-m-Bz); 7.62, 7.678, 7.684 (3m, 31H, H-p-Bz); 7.81, 7.97, 8.03 (3m, 32H, H-o-Bz); 8.20 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 39.36 ((CH.sub.3).sub.2N); 63.91 (CH.sub.2-5); 70.88 (CH-3); 71.79 (CH-2); 79.17 (CH-4); 85.59 (CH-1); 97.93 (C-4a); 119.86 (CH-6); 121.21 (CH-5); 122.04 (C-4b); 128.37, 128.79 (C-i-Bz); 128.96, 129.04 (CH-m-Bz); 129.46 (C-i-Bz); 129.49, 129.54, 129.60 (CH-o-Bz); 133.80, 134.15, 134.19 (CH-p-Bz); 135.17 (C-7a); 151.12 (CH-2); 156.29 (C-8a); 157.01 (C-4); 164.59, 164.97, 165.71 (CO-Bz). ESI MS m/z (rel %): 663 (15) [M+H]. 685 (100) [M+Na]. HR MS (ESI) for C.sub.36H.sub.31N.sub.4O.sub.7S [M+H]: calcd 663.19080; found 663.19088.

Example 10

4-(2-Furyl)-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1a)

(74) Compound 7a (140 mg, 0.20 mmol) was deprotected according to the general procedure A. Nucleoside 1a (52 mg, 68%) was obtained as a white solid. m.p. 189-192 C. [].sub.D 65.1 (c 0.19). IR (ATR): v=3242, 2930, 1579, 1548, 1504, 1420, 1401, 1331, 1127, 1099, 1041, 719. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.63, 3.67 (2bdd, 21H, J.sub.gem=11.5, J.sub.5,4=5.2, H-5); 3.98 (td, 1H, J.sub.4,5=5.2, J.sub.4,3=2.8, H-4); 4.13 (dd, 1H, J.sub.3,2=5.4, J.sub.3,4=2.8, H-3); 4.60 (dd, 1H, J.sub.2,1=7.1, J.sub.2,3=5.4, H-2); 5.01 (bs, 1H, OH-5); 5.35 (bs, 1H, OH-3); 5.49 (bs, 1H, OH-2); 6.48 (d, 1H, J.sub.1,2=7.1, H-1); 6.85 (dd, 1H, J.sub.4,3=3.5, J.sub.4,5=1.7, H-4-furyl); 7.43 (d, 1H, J.sub.6,5=5.4, H-6); 7.50 (dd, 1H, J.sub.3,4=3.5, J.sub.3,5=0.8, H-3-furyl); 7.96 (d, 1H, J.sub.5,6=5.4, H-5); 8.25 (dd, 1H, J.sub.5,4=1.7, J.sub.5,3=0.8, H-5-furyl); 8.83 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 62.19 (CH.sub.2-5); 70.76 (CH-3); 71.11 (CH-2); 85.47 (CH-4); 86.45 (CH-1); 106.96 (C-4a); 112.97 (CH-4-furyl); 113.41 (CH-3-furyl); 121.09 (CH-5); 121.38 (CH-6); 121.41 (C-4b); 140.88 (C-7a); 145.00 (C-4); 146.68 (CH-5-furyl); 151.14 (C-2-furyl); 152.55 (CH-2); 157.45 (C-8a). ESI MS m/z (rel %): 396 (100) [M+Na]. HR MS (ESI) for C.sub.17H.sub.15N.sub.3O.sub.5SNa [M+Na]: calcd 396.06246; found 396.06251.

Example 11

4-(3-Furyl)-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1b)

(75) Crude compound 7b (300 mg, 70% purity, 0.31 mmol) was deprotected according to the general procedure A. Nucleoside 1b (95 mg, 83%) was obtained as white crystals. m.p. 192-195 C. [].sub.D 1.6 (c 0.19, DMSO). IR (ATR): v=3424, 3225, 3161, 1565, 1496, 1453, 1441, 1301, 1261, 1132, 1051, 1016, 878, 817, 795, 652, 643, 596. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.62, 3.67 (2ddd, 21H, J.sub.gem=11.5, J.sub.5,OH=5.5, J.sub.5,4=5.2, H-5); 3.98 (td, 1H, J.sub.4,5=5.2, J.sub.4,3=2.8, H-4); 4.12 (ddd, 1H, J.sub.3,2=5.5, J.sub.3,OH=4.6, J.sub.3,4=2.8, H-3); 4.59 (ddd, 1H, J.sub.2,1=7.3, J.sub.2,OH=6.3, J.sub.2,3=5.5, H-2); 5.04 (t, 1H, J.sub.OH,5=5.5, OH-5); 5.35 (d, 1H, J.sub.OH,3=4.6, OH-3); 5.50 (d, 1H, J.sub.OH,2=6.3, OH-2); 6.48 (d, 1H, J.sub.1,2=7.3, H-1); 7.19 (dd, 1H, J.sub.4,5=1.9, J.sub.4,2=0.9, H-4-furyl); 7.43 (d, 1H, J.sub.6,5=5.4, H-6); 7.62 (d, 1H, J.sub.5,6=5.4, H-5); 7.96 (dd, 1H, J.sub.5,4=1.9, J.sub.5,2=1.6, H-5-furyl); 8.57 (dd, 1H, J.sub.2,5=1.6, J.sub.2,4=0.9, H-2-furyl); 8.86 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 62.24 (CH.sub.2-5); 70.82 (CH-3); 71.22 (CH-2); 85.54 (CH-4); 86.54 (CH-1); 109.79 (C-4a); 110.32 (CH-4-furyl); 119.50 (CH-5); 120.69 (C-4b); 121.74 (CH-6); 125.17 (C-3-furyl); 140.65 (C-7a); 144.42 (CH-2-furyl); 144.90 (CH-5-furyl); 149.34 (C-4); 151.29 (CH-2); 156.92 (C-8a). ESI MS m/z (rel %): 396 (100) [M+Na]. HR MS (ESI) for C.sub.17H.sub.15N.sub.3O.sub.5SNa [M+Na]: calcd 396.06246; found 396.06237.

Example 12

4-(2-Benzofuryl)-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1c)

(76) Compound 7c was deprotected according to the general procedure C. Nucleoside 1c (100 mg, 86%) was obtained as yellowish crystals. m.p. 118-119 C. [].sub.D 29.6 (c 0.13). IR (ATR): v=3259, 1563, 1494, 1435, 1300, 1276, 1129, 1054, 1044, 1025, 794, 744, 644, 599. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.65, 3.69 (2ddd, 21H, J.sub.gem=11.5, J.sub.5,OH=5.5, J.sub.5,4=5.2, H-5); 3.99 (td, 1H, J.sub.4,5=5.2, J.sub.4,3=2.8, H-4); 4.14 (ddd, 1H, J.sub.3,2=5.3, J.sub.3,OH=4.7, J.sub.3,4=2.8, H-3); 4.62 (ddd, 1H, J.sub.2, 1=7.1, J.sub.2,OH=6.3, J.sub.2,3=5.3, H-2); 5.03 (t, 1H, J.sub.OH,5=5.5, OH-5); 5.36 (d, 1H, J.sub.OH,3=4.7, OH-3); 5.52 (d, 1H, J.sub.OH,2=6.3, OH-2); 6.52 (d, 1H, J.sub.1,2=7.1, H-1); 7.40 (ddd, 1H, J.sub.5,4=7.9, J.sub.5,6=7.2, J.sub.5,7=1.0, H-5-benzofuryl); 7.50 (d, 1H, J.sub.6,5=5.4, H-6); 7.52 (ddd, 1H, J.sub.6,7=8.4, J.sub.6,5=7.2, J.sub.6,4=1.3, H-6-benzofuryl); 7.86 (ddd, 1H, J.sub.4,5=7.9, J.sub.4,6=1.3, J.sub.4,7=1.0, H-4-benzofuryl); 7.95 (d, 1H, J.sub.3,7=1.0, H-3-benzofuryl); 8.00 (dq, 1H, J.sub.7,6=8.4, J.sub.7,3=J.sub.7,4=J.sub.7,5=1.0, H-7-benzofuryl); 8.12 (d, 1H, J.sub.5,6=5.4, H-5); 8.94 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 62.18 (CH.sub.2-5); 70.77 (CH-3); 71.16 (CH-2); 85.55 (CH-4); 86.52 (CH-1); 108.22 (C-4a); 109.12 (CH-3-benzofuryl); 112.07 (CH-7-benzofuryl); 121.27 (C-4b); 121.60 (CH-5,6); 122.72 (CH-4-benzofuryl); 124.20 (CH-5-benzofuryl); 126.73 (CH-6-benzofuryl); 127.88 (C-3a-benzofuryl); 141.64 (C-7a); 144.75 (C-4); 151.15 (CH-2); 154.19 (C-2-benzofuryl); 155.58 (C-7a-benzofuryl); 157.68 (C-8a). ESI MS m/z (rel %): 424 (31) [M+H]; 446 (100) [M+Na]. HR MS (ESI) for C.sub.21H.sub.17N.sub.3O.sub.5S [M+H]: calcd 423.0889; found 423.0894.

Example 13

4-Methyl-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1d)

(77) Crude compound 7d (125 mg, 0.23 mmol) was deprotected according to the general procedure A. Nucleoside 1d (43 mg, 70%) was obtained as a white lyophilizate (water/tBuOH). m.p. 126-128 C. [].sub.D 62.3 (c 0.20). IR (ATR): v=3524, 3131, 2848, 1608, 1504, 1450, 1402, 1323, 1257, 1113, 1051, 654. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 2.84 (s, 3H, CH.sub.3); 3.58-3.67 (bm, 2H, H-5); 3.95 (td, 1H, J.sub.4,5=5.1, J.sub.4,3=2.8, H-4); 4.11 (dd, 1H, J.sub.3,2=5.3, J.sub.3,4=2.8, H-3); 4.56 (dd, 1H, J.sub.2,1=7.2, J.sub.2,3=5.3, H-2); 5.01 (bs, 1H, OH-5); 5.34 (bs, 1H, OH-3); 5.47 (bs, 1H, OH-2); 6.41 (d, 1H, J.sub.1,2=7.2, H-1); 7.42 (d, 1H, J.sub.6,5=5.3, H-6); 7.63 (d, 1H, J.sub.5,6=5.3, H-5); 8.73 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 22.28 (CH.sub.3); 62.22 (CH.sub.2-5); 70.80 (CH-3); 71.24 (CH-2); 85.42 (CH-4); 86.42 (CH-1); 112.20 (C-4a); 118.75 (CH-5); 121.52 (C-4b); 121.83 (CH-6); 139.32 (C-7a); 151.25 (CH-2); 155.73 (C-8a); 157.35 (C-4). ESI MS m/z (rel %): 322 (25) [M+H]; 344 (100) [M+Na]. HR MS (ESI) for C.sub.14H.sub.15N.sub.3O.sub.4S [M+H]: calcd 321.0783; found 321.0789.

Example 14

4-N,N-dimethylamino-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1e)

(78) Compound 7e (90 mg, 0.14 mmol) was deprotected according to the general procedure A. Nucleoside 1e (42 mg, 88%) was obtained as white crystals. m.p. 117 C. [].sub.D 8.5 (c 0.16). IR (ATR): v=3258, 1583, 1460, 1442, 1420, 1314, 1112, 1053, 787, 646. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.34 (s, 6H, (CH.sub.3).sub.2N); 3.59, 3.64 (2dd, 21H, J.sub.gem=11.6, J.sub.5,4=5.2, H-5); 3.92 (td, 1H, J.sub.4,5=5.2, J.sub.4,3=3.0, H-4); 4.08 (dd, 1H, J.sub.3,2=5.5, J.sub.3,4=3.0, H-3); 4.55 (dd, 1H, J.sub.2,1=7.1, J.sub.2,3=5.5, H-2); 5.07, 5.37 (2bs, 3H, OH-2,3,5); 6.35 (d, 1H, J.sub.1,2=7.1, H-1); 7.27 (d, 1H, J.sub.6,5=5.5, H-6); 7.44 (d, 1H, J.sub.5,6=5.5, H-5); 8.23 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 39.39 ((CH.sub.3).sub.2N); 62.29 (CH.sub.2-5); 70.82 (CH-3); 71.00 (CH-2); 85.20 (CH-4); 86.80 (CH-1); 97.76 (C-4a); 119.65 (CH-6); 120.88 (CH-5); 121.53 (C-4b); 135.58 (C-7a); 150.85 (CH-2); 156.55 (C-8a); 157.10 (C-4). ESI MS m/z (rel %): 351 (39) [M+H]; 373 (100) [M+Na]. HR MS (ESI) for C.sub.15H.sub.19N.sub.4O.sub.4S [M+H]: calcd 351.11215; found 351.11223.

Example 15

4-Amino-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1f)

(79) Nucleoside 6 (280 mg, 0.42 mmol) was dissolved in dioxane (4 ml) and 30% aq. ammonia (12 ml) was added. The reaction mixture was heated in pressure glass vial at 100 C. for 24 hr, cooled and solvents were evaporated. Purification by RP-HPFC (water/methanol, 10.fwdarw.100%) gave compound 1f (107 mg, 78%) as white crystals. m.p. 98 C. [].sub.D 24.7 (c 0.15). IR (ATR): v=3452, 3347, 3073, 2933, 2862, 1723, 1605, 1551, 1453, 1263, 1093, 1068, 1028, 707. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.58, 3.63 (2bdd, 21H, J.sub.gem=11.7, J.sub.5,4=5.0, H-5); 3.92 (td, 1H, J.sub.4,5=5.0, J.sub.4,3=2.8, H-4); 4.08 (ddd, 1H, J.sub.3,2=5.5, J.sub.3,OH=4.3, J.sub.3,4=2.8, H-3); 4.55 (ddd, 1H, J.sub.2,1=7.1, J.sub.2,OH=6.5, J.sub.2,3=5.5, H-2); 5.08 (bs, 1H, OH-5); 5.25 (bd, 1H, J.sub.OH,3=4.3, OH-3); 5.40 (bd, 1H, J.sub.OH,2=6.5, OH-2); 6.26 (d, 1H, J.sub.1,2=7.1, H-1); 7.18 (bs, 2H, NH.sub.2); 7.26 (d, 1H, J.sub.6,5=5.3, H-6); 7.82 (d, 1H, J.sub.5,6=5.3, H-5); 8.16 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 62.33 (CH.sub.2-5); 70.89 (CH-3); 71.22 (CH-2); 85.22 (CH-4); 86.77 (CH-1); 96.80 (C-4a); 118.91 (CH-6); 119.96 (CH-5); 121.48 (C-4b); 135.57 (C-7a); 151.95 (CH-2); 155.89 (C-8a); 156.19 (C-4). ESI MS m/z (rel %): 323 (15) [M+H]; 345 (100) [M+Na]. HR MS (ESI) for C.sub.13H.sub.15N.sub.4O.sub.7S [M+H]: calcd 323.08085; found 323.08091.

Example 16

4-Methoxy-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1g)

(80) Nucleoside 6 (130 mg, 0.20 mmol) was suspended in methanol (20 ml) and sodium methoxide (15 mg, 0.26 mmol) was added. The reaction mixture was stirred overnight at r.t., solvent was evaporated and crude material was purified by RP-HPFC chromatography (water/methanol 10.fwdarw.100%). Nucleoside 1g (43 mg, 65%) was obtained as white crystals. m.p. 159-160 C. [].sub.D 47.3 (c 0.15, DMSO). IR (ATR): v=3617, 3480, 2951, 1610, 1564, 1443, 1335, 1308, 1205, 1127, 1052, 1023, 975, 635, 602. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 3.60, 3.64 (2ddd, 21H, J.sub.gem=11.6, J.sub.5,OH=5.6, J.sub.5,4=5.1, H-5); 3.95 (td, 1H, J.sub.4,5=5.1, J.sub.4,3=2.8, H-4); 4.10 (dd, 1H, J.sub.3,2=5.3, J.sub.3,OH=4.7, J.sub.3,4=2.8, H-3); 4.13 (s, 3H, CH.sub.3O); 4.55 (dd, 1H, J.sub.2,1=7.1, J.sub.2,OH=6.4, J.sub.2,3=5.3, H-2); 5.01 (t, 1H, J.sub.OH,5=5.6, OH-5); 5.32 (d, 1H, J.sub.OH,3=4.7, OH-3); 5.47 (d, 1H, J.sub.OH,2=6.4, OH-2); 6.38 (d, 1H, J.sub.1,2=7.1, H-1); 7.38 (d, 1H, J.sub.5,6=5.3, H-5); 7.39 (d, 1H, J.sub.6,5=5.3, H-6); 8.53 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 54.11 (CH.sub.3O); 62.22 (CH.sub.2-5); 70.80 (CH-3); 71.29 (CH-2); 85.42 (CH-4); 86.72 (CH-1); 99.35 (C-4a); 118.11 (CH-5); 120.51 (C-4b); 122.00 (CH-6); 137.53 (C-7a); 151.17 (CH-2); 157.17 (C-8a); 161.53 (C-4). ESI MS m/z (rel %): 360 (100) [M+Na]. HR MS (ESI) for C.sub.14H.sub.15N.sub.3O.sub.5SNa [M+Na]: calcd 360.06246; found 360.06254.

Example 17

4-Methylsulfanyl-8-(-D-ribofuranosyl)-8H-thieno[3,2:4,5]pyrrolo[2,3-d]pyrimidine (1h)

(81) Nucleoside 6 (80 mg, 0.12 mmol) was suspended in methanol (10 ml) and sodium thiomethoxide (12 mg, 0.16 mmol) was added. The reaction mixture was stirred overnight at r.t., solvent was evaporated and crude material was purified by RP-HPFC chromatography (water/methanol 10.fwdarw.100%). Nucleoside 1h (33 mg, 90%) was obtained as white crystals. m.p. 148-152 C. [].sub.D 50.9 (c 0.16). IR (ATR): v=3305, 1576, 1556, 1497, 1471, 1431, 1321, 1265, 1246, 1136, 1094, 1055, 912, 821, 719, 645. .sup.1H NMR (500.0 MHz, DMSO-d.sub.6): 2.74 (s, 3H, CH.sub.3S); 3.61, 3.64 (2dd, 21H, J.sub.gem=11.6, J.sub.5,4=5.2, H-5); 3.96 (td, 1H, J.sub.4,5=5.2, J.sub.4,3=2.8, H-4); 4.11 (dd, 1H, J.sub.3,2=5.3, J.sub.3,4=2.8, H-3); 4.55 (dd, 1H, J.sub.2,1=7.1, J.sub.2,3=5.3, H-2); 4.90-5.70 (bm, 3H, OH-2,3,5); 6.38 (d, 1H, J.sub.1,2=7.1, H-1); 7.43 (d, 1H, J.sub.5,6=5.3, H-5); 7.45 (d, 1H, J.sub.6,5=5.3, H-6); 8.73 (s, 1H, H-2). .sup.13C NMR (125.7 MHz, DMSO-d.sub.6): 11.57 (CH.sub.3S); 62.19 (CH.sub.2-5); 70.77 (CH-3); 71.33 (CH-2); 85.49 (CH-4); 86.54 (CH-1); 110.05 (C-4a); 118.07 (CH-5); 120.83 (C-4b); 122.46 (CH-6); 138.67 (C-7a); 151.00 (CH-2); 153.70 (C-8a); 159.00 (C-4); ESI MS m/z (rel %): 376 (100) [M+Na]. HR MS (ESI) for C.sub.14H.sub.16N.sub.3O.sub.4S.sub.2 [M+H]: calcd 354.05767; found 354.05772.

(82) In Vitro Antitumor Activity

(83) MTT test was used for in vitro evaluation of antitumor activities of newly synthesized compounds on cell lines derived from normal tissues or tumors. Specifically, cell lines K562 (human acute myeloid leukemia), K562-Tax (human acute myeloid leukemia, paclitaxel resistant subline, overexpress multiple drug resistant protein PgP), CEM (T-lymfoblastic leukemia), CEM-DNR-bulk (T-lymfoblastic leukemia, doxorubicin resistant), A549 (human lung adenocarcinoma), HCT116 wt (human colorectal cancer, wild-type), HCT116p53/(human colorectal cancer, mutant p53) a U2OS (human bone osteosarcoma) were used.

(84) Express characteristics, susceptibility profiles of classic antitumor drugs as well as methodology of cytotoxic MTT test have been repeatedly published {ref.:Noskova, V.; Dzubak, P.; Kuzmina, G.; Ludkova, A.; Stehlik, D.; Trojanec, R.; Janostakova, A.; Korinkova, G.; Mihal, V.; Hajduch, M., Neoplasma 2002, 49, 418-425}.

(85) Results of Biological Testing:

(86) The tested compounds showed activity in in vitro cytotoxic test (Table 4), and it was selective against broad spectrum of cancer cell lines of various histogenetic origin (mesenchymal or epitelial tumors) with significantly lower activity against normal human fibroblasts (BJ and MRC-5 cell lines were tested). Active compounds showed promising therapeutic indexes (15-2500). IC.sub.50 values of compounds 1c, if were in micromolar range, IC.sub.50 values of compounds 1d, 1g, 1h were sub-micromolar to nanomolar. Cytotoxic activity against cancer cells was independent on p53 gene status, same activities were found for HCT116 (p53 wild type) and for mutant line with deleted gene HCT116 (p53 /). However, a number of derivatives showed lower cytotoxicity against cells overexpressing transport proteins (mdr-1 for K562-TAX line and mrp-1 for CEM-DNR).

(87) TABLE-US-00003 TABLE 4 Cytotoxic activities of prepared compounds Compound A549 CCRF-CEM CEM-DNR HCT116 HCT116p53 K562 K562-TAX U2OS 1a E E E E E E E E 1b E E E E E E E E 1c E D D E E E D E 1d B A E B B A C A 1e E E E E E E E E 1f E B E E C E B B 1g C A E B B B D A 1h B A E A B B D A IC.sub.50: A = 10-200 nmol .Math. l.sup.1; B = 200-900 nmol .Math. l.sup.1; C = 0.9-10 mol .Math. l.sup.1; D = 10-25 mol .Math. l.sup.1; E = 25-50 mol .Math. l.sup.1.

INDUSTRIAL APPLICABILITY

(88) In particular, the compounds of this invention can be used as medicaments or components of medicaments for treatment of cancer and leukemia.