DE19514523A1 - New cytosine and cytidine derivatives - Google Patents

Abstract

The invention concerns cytosine and cytidine derivatives of general formula (1) and their production and use in medicaments.

Description

The invention relates to cytosine and cytidine derivatives, their preparation and use in Medicines.

The therapeutic use of cytidines, especially aracytidin and others Cytosine derivatives as cytostatics or antivirals are affected by their rapid deactivation by cytidine deaminases, being from the biologically potent Cytidines or cytosine derivatives the corresponding inactive uridines or uracil derivatives arise as by G. W. Camiener et al., Biochem. Pharmacol 14, 1405 (1965), 16, 1681 (1967) and W. Kreis et al., Helv. Chim. Acta 61, 1011 (1978). Therefore, it is of great therapeutic interest to synthesize cytidines or cytosine derivatives that are not substrates of cytidine deaminase and have a longer duration of action in vivo.

It has now surprisingly been found that Ansa-cytosine and Ansa-cytidine derivatives which a dimethylene grouping between the C⁵ carbon atom and the N⁴ nitrogen Have atoms, represent biologically potent molecules that are not substrates of the cytidine Are deaminase and therefore do not have the disadvantages mentioned above of the state of the art Technology-owned cytosine and cytidine derivatives.

The invention relates to cytosine and cytidine derivatives of the general formula 1,

wherein
R is hydrogen, CHO, COR⁶ with R⁶ in the meaning of C₁-C₁₇-alkyl and phenyl, or COOR⁷ with R⁷ in the meaning of C₁-C₄-alkyl or benzyl
R ′ is hydrogen or the groups

wherein
n 1 or 2
XO, S, CH₂ or NR,
R² is hydrogen, fluorine, CH₃ or CN,
R³ is hydrogen, fluorine, hydroxy or -OCH₃,
R⁴ is hydrogen, fluorine, hydroxy, N₃ or NH₂,
R⁵ is hydrogen or PO (OH) ₂,
R², R³ together = CH₂,
R³, R⁴ together form a double bond and their salts.

If n = 2, all substituents R² and R³ can be different.

The compounds of general formula 1 can be represented, for example, as follows put:

The well-known 5-hydroxyethylcytosine 7 (J.D. Fissekis et al .; J. Org. Chem. 29, 2670 (1964)) can z. B. with triphenylphosphine dihalides such as bromide or chloride and Triethylamine or after Mitsunobu to the new, not yet described Ansa-Cytosin 1 (R = R₁ = H) cyclize, which leads to the N-acyl derivatives such as the N⁴-acetyl derivative 1 (R = COCH₃; R '= H) can be acylated. Silylation of 1 (R = R₁ = H) with hexamethyldisilazane (HMDS) in the presence of a catalytic amount of trimethylchlorosilane (TCS) in abs. Acetonitrile gives the 2,4-bistrimethylsilyl compound 8a, which deals with 1-O-acetyl-2,3,5-tri-O- benzoyl-β-D-ribofuranose 9 in the presence of trimethylsilyl triflate (or SnCl₄) in Solvents such as abs. 1,2-dichloroethane or acetonitrile in the protected crystalline Ansacytidine 10 can be converted. The monosilylated N⁴-acetyl compound 8b reacts analogously to the N⁴-acetylated 2 ′, 3 ′, 5′-tri-O-benzoyl-ribofuranoside (10 with N⁴-acetyl group). Saponification of the tribenzoate 10 gives the free, crystalline ansacytidine 11. Alternatively, it provides  the reaction of persilylated 5-hydroxyethyluracil with 1-O-acetyl-2,3,5-tri-O-benzoyl β-D-ribose 9 in the presence of trimethylsilyl triflate (or SnCl₄) the 5-hydroxyethyl-2 ′, 3 ′, 5′- tri-O-benzoyl-uridine 12. Reaction of 12 with POCl₃ (or with triphenylphosphine / 1,2-di bromotetrachloroethane) in acetonitrile gives the protected dichloro (or di-bromo) compound 13, which deals with methanolic ammonia in the presence of tertiary bases such as Triethylamine, ethyldiisopropylamine or DBU with simultaneous saponification of the O- Benzoyl groups cyclized to free ansacytidine 11.

a) R⁵ = H, b) R⁵ = PO (OH) ₂, c) R⁵ = PO (OH) ONa

The Ansacytidin 11 can by the usual methods, for. B. by reaction of 11 with 2-acetoxyisobutyric acid chloride (J.G. Moffatt et al .; J. Org. Chem. 39, 2182 (1974)) or 2- Acetoxybenzoyl chloride (J.J. Fox et al .; Synthesis 533 (1976)) at 24 ° C in acetonitrile or by heating with diphenyl carbonate in the presence of sodium bicarbonate (C. B. Reese et al., J. Chem. Soc., Perkin Trans. I, 1172 (1982)), or ethylene carbonate (Ajinomoto Inc., DOS 2.261215) into 2,2'-anhydro-ansacytidine hydrochloride 14. Subsequent treatment treatment with aqueous ammonia or 1 N NaOH, evaporation, and crystallization Methanol provides the crystalline Ansa-ara-cytidine 15a. Implementation with equivalent amounts methanolic HCl and careful concentration at T <20 ° gives the crystalline solution  ara-cytidine hydrochloride 15a HCl. Phosphorization with POCl₃ in triethyl phosphate results the phosphate 15b from which the monophosphate 15c can be produced overnight.

Alternatively, 2,2'-anhydro-ansacytidine 14 from the oxazoline 16, that from D-arabinose and cyanamide is easily accessible (R.A. Sanchez and L.E. Orgel, J. Mol Biol. 47, 531 (1970); D.H. Shannahoff and R.A. Sanchez, J. Org. Chem. 38, 593 (1973)), by Kon Prepare the seal with 2-formyl-pyrrolidone 17. Condensation of 17 with urea or N¹, N²-bis-trimethylsilylurea gives ansacytosine 1 (R = H; DPTBS; BOC).

Selective protection of the 5′-hydroxyl group in 11 (or the corresponding N⁴-acetyl derivative) by tritylation or silylation, subsequent mesylation of the 2 ', 3'-hydroxyl groups and treatment with Li₂Te provides the 5′-protected 2 ′, 3′-didehydro-2 ′, 3′-dideoxy compound, from which the 2 ', 3'- by hydrogenation and subsequent removal of the 5'-protecting group Dideoxyansa-ara-cytidine can be produced. Reaction of the silyl compound 8b with 1-α-chloro-2- deoxy-3,5-di-O-toluolyl-D-ribofuranose 18 in the presence of trimethylsilyl triflate in abs. 1,2-dichloroethane gives an α / β mixture of the protected 2'-deoxy compounds from which the β-anomer 19 can be separated from the α-anomer 20 in crystalline form.

Saponification of 19 with methanolic ammonia leads to the free crystalline 2'-deoxy ansacytidine 21

Reaction of 8b with the 2-fluorine derivative 22 (see J. A. Martin et al., J. Med. Chem. 33, 2137 (1990)) followed by saponification and separation results in addition to the β-anomer 23 and the α-anomer 24th

Reaction of 8b with the 2,2-difluorosugar 25 (T. S. Chou, P. C. Heath, L. E. Patterson, L.M. Poket, R.E. Lakin and A.H. Hunt, Synthesis 565 (1992)) in the presence of Trime After separation and saponification, thylsilyl triflate in 1,2-dichloroethane gives the 2′-deoxy-2 ′, 2′- difluoro-ansacytidine 26 and the corresponding α-anomer 27.

Reaction of 2'-deoxyansacytidine 21 with tert-butyldiphenylsilyl chloride and follow heating with diphenyl carbonate / NaHCO₃ gives 2'-deoxy-2,3'-anhydro-ansa-cytidine  28, the one with HN₃ or AlF₃ and subsequent separation of the TBDPS group with F⁻ in the 3'-azido derivative 29 and the 3'-fluoro derivative 30 can be converted.

The 3'-azido derivative 29 can be hydrogenated to the 3'-amino derivative. Reaction of the N-acetylated Ansacytidins 31 with TIPS-Cl₂ and subsequent CrO₃-oxidation gives the 2'-ketone 32 that the free 2′-methylene nucleoside 33 in a Wittig reaction followed by fluoride treatment delivers. Reaction of ketone 22 with NaCN / NaHCO₃ in Et₂O / H₂O leads to cyanohydrin 34. Acylation of 34 with C₆H₅-O-CS-Cl in the presence of triethylamine-DMAP and after following radical reduction with Bu₃SnH / AIBN and finally cleavage of the silyl The 2′-deoxy-2-cyanoansacytidine 35 provides groups (cf. A. Matsuda et al., Tetrahedron Lett. 32, 6003 (1991)).

The implementation of 8b with reactive intermediates such. B. the corresponding halo gender derivatives gives the Secoderivate 3 ′ (see A. Holy, Coll Czech. Commun. 58, 649 (1993), P.R. Brodfuehrer et al., Tetrahedron Lett. 35, 3243 (1994)), 4 ′ and 5 ′, while the

Reaction of 8 with 5-acetoxy-2-acetoxymethylthiazolidine in the presence of trimethyl silyl triflate and subsequent saponification provides a cis / trans mixture from which the Pure nucleoside 6 can be separated.

The invention relates to the process for the preparation of the compounds of the formula 1

wherein the radicals R ′ and R have the meaning given above, characterized in that that he

• a) a compound of formula 7 with triphenylphosphine dihalide and a tertiary amine or
• b) a formylpyrrolidone derivative of the formula 17 in which R¹³ is hydrogen, a SiR¹⁴R¹⁵R¹⁶ radical in which R¹⁴, R¹⁵, R¹⁶ are identical or different and are phenyl or C₁-C₄-alkyl, or is tert-butyloxycarbonyl,
  with a urea derivative or an isourea derivative such as 16,
  wherein R¹⁷, R¹⁸ are the same or different and are hydrogen or SiR¹⁴R¹⁵R¹⁶,

implemented and, if desired, then formylated, acylated or introduced in the 7-position or
in the 3-position introduces the optionally hydroxyl-containing radical R 'and then, if desired, introduces N₃, NH₂, fluorine, CN or = CH₂ or R³, R⁴ in the meaning of a double bond and if desired hydrogenates or 5' hydroxyl groups in the 5'-phosphate transferred and, if necessary, splitting off protective groups or forming the salts.

Alkali salts such as sodium and potassium salts and ammonium salts are suitable as salts of the 5′-phosphates. The customary groups can be used as hydroxyl protective groups. Alkyl is to be understood as straight-chain and branched alkyls, in particular C _1-4 alkyls and C₁₅ and C₁₆ alkylene radicals.

The preferred meaning of the radical R 'is group 2, in particular n = 1 and X = O are preferred.

The nucleosides of formula 1 with free 5'-hydroxyl groups can be known Methods (M. Yoshikawa, T. Kato and T. Takenishi, Bull Chem. Soc. Jap. 42, 3505 (1969) and T. Sowa and S. Ouchi, Bull Chem. Soc. Yep 48, 2084 (1975)) transfer the corresponding 5'-phosphates and their salts.

The listed new ansacytidines of general formula 1 and their 5'-phosphates and Salts are particularly characterized by their long-lasting in vitro and in vivo action different mouse leukemia models, especially in mouse L1210 leukemia, the HL 60 cell culture and various viral diseases.

Compound 21 and the corresponding phosphoramidates of 21 are particularly suitable are excellent for the formation of stable triplex DNA (cf. 5-methyl-2′deoxycytidine: G.C. Best and P.B. Dervan, J. Amer. Chem. Soc. 117; 1187 (1995)).

For the use of the compounds according to the invention as pharmaceuticals, these are Form of a pharmaceutical preparation, which in addition to the active ingredient for enteral or parenteral administration of suitable pharmaceutical, organic or inorganic inert Carrier materials, such as water, gelatin, gum arabic, milk sugar, Contains starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols etc. The  pharmaceutical preparations can be in solid form, for example as tablets, coated tablets, Suppositories, capsules or in liquid form, for example as solutions, suspensions or emulsions are present. If necessary, they also contain auxiliary substances such as Preservatives, stabilizers, wetting agents or emulsifiers, salts for changing the osmotic pressure or buffer.

For parenteral use, especially injection solutions or suspensions, in particular aqueous solutions of the active compounds in polyhydroxyethoxylated Castor oil, suitable.

Surface-active auxiliaries such as salts of bile acids can also be used as carrier systems or animal or vegetable phospholipids, but also mixtures thereof and liposomes or their components are used.

Tablets, coated tablets or capsules with talc are particularly suitable for oral use and / or hydrocarbon carriers or binders, such as lactose, maize or Potato starch, suitable. The application can also be in liquid form, such as Example as juice, to which a sweetener may be added.

The dosage of the active ingredients can vary depending on the route of administration, age and weight of the Patients, type and severity of the disease to be treated and similar factors vary.

Insofar as the preparation of the starting compounds is not described, they are known or can be prepared analogously to known compounds or processes described here.

The following examples are intended to explain the process according to the invention:

example 1 Preparation of 3,5,6,7-tetrahydro-2H-pyrrolo [2,3d] pyrimidin-2-one 1 (R = R ′ = H)

To a stirred suspension of 7.75 g (50 mmol) of the known 5- (2-hydroxyethyl) cytosine 7 (JD Fissekis et al., J. Org. Chem. 29, 2670 (1964)) and 15.73 g (60 mmol) triphenylphosphine was at +4 → 7 ° a solution of 17.91 g (55 mmol) 1,2-dibromotetrachloroethane in 75 ml abs. DMF added dropwise within 1 h and stirred for a further 0.5 h at + 6 °. After warming up to 21 °, the reaction mixture was left at 21 ° C. after 18 h, then 25.85 g (200 mmol) of N-ethyl-N-diisopropylamine were added and the reaction mixture was heated at an oil bath temperature of 82-84 ° for 2 h, initially a clear solution emerged, from which crystals precipitated again after about 45 minutes. After cooling to 65 °, the mixture was filtered and the extracted substance was washed with 35 ml of DMF, 2.51 g of pure 1 being obtained. The filtrate was evaporated at 65-70 ° in vacuo and the residue was stirred with 500 ml of CH₂Cl₂ and 20 ml of N-ethyl-N-diisopropylamine and the undissolved substance was suctioned off and again with 100 ml of CH₂Cl₂ and 10 ml of N-ethyl-N-diisopro washed pylamine and dried, whereby a further 5.03 g 1 were obtained. Both positions 1 were suspended in 150 ml of H₂O and sat so long with stirring. NaHCO₃ solution added until ap _H = 8-9 was reached. After stirring for a further hour at 24 °, the mixture was evaporated and the residue was recrystallized from 50 ml of H₂O with the addition of activated carbon, giving 4.8 g of 1, mp. 310 ° (decomp.) (Sublimation at 260 °) (¹H- NMR (DMSO-D₆ δ: 2.68-2.73 (tr, CH₂-C), 3.56-3.63 (tr, CH₂-N), 7.18 (s, HC = C).

Example 2 Preparation of 7-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one 1 (R = COCH₃; R ′ = H)

3.7 g (29.98 mmol) of 3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one were added in 50 ml Section. Suspended pyridine and added 8 ml of acetic anhydride, whereupon the Reaction mixture heated to 28 °. The solution that became clear after stirring for one hour was evaporated in vacuo after 18 h, three times after the addition of 30 ml of abs. Xylene concentrated and the residue dried on the oil pump. When recrystallizing from 50 ml Methanol and 2 ml of H₂O were 1.58 g of N-acetyl compound, mp. 274-279 ° and at Concentration of the mother liquor to 25 ml a further 0.68 g = a total of 2.26 g of N- Acetyl compound obtained. The evaporated mother liquor was treated with 25 ml pyridine - 5 ml Acetanhydrid nachacetyliert and after 20 h with xylene several times in vac. evaporated, being a further 2.1 g of practically pure N-acetyl compound were obtained. Mp 274-279 ° C, ¹H-  NMR (DMSO-D₆) δ: 2.56 (s, CH₃CO), 2.72-2.78 (m, CH₂-C =), 3.88-3.92 (m, CH₂-N), 7.56 (s, H-C = C), 11.05 (br, HN)

Example 3 3- (2 ′, 3 ′, 5′-tri-O-benzoyl-β-D-ribofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3- d] pyrimidin-2-one 10

Approx. 5-6 g of crude 3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one 1 (R ′ = R = H) were with 20 ml abs. Pyridine boiled with 125 ml of hexamethyldisilazane for 12 h, then in vacuo concentrated and the residue evaporated twice with 50 ml of xylene and finally on the Oil pump dried to give 10.41 (36.97 mmol) of crystalline bisilyl compound 8a has been. The silyl compound was treated with 18.64 g (36.97 mmol) of 1-O-acetyl-2,3,5-tri-O- benzoyl-β-D-ribofuranose 9 under nitrogen in 200 ml abs. Dissolved 1,2-dichloroethane, then 8th ml (44 mmol) trimethylsilyl triflate in 75 ml 1,2-dichloroethane within 30 min Stir added dropwise at 24 ° and leave the reaction mixture at 24 ° for 72 h. To Dilution with 250 ml CH₂Cl₂ was with 200 ml ice-cold saturated NaHCO₃ solution shaken, the phases separated and the aqueous phase re-extracted with 2 × 200 ml CH₂Cl₂. After drying (Na₂SO₄) and evaporation, the amorphous residue (22.4 g) CH₂Cl₂ a column of 450 g SiO₂ chromatographed, 1 l CH₂Cl₂ and another 2 l CH₂Cl₂- Isopropanol (98: 2) contained only 3.15 g of by-products, while further elution with 2-3 l of the 98: 2 mixture gave 17.25 g (80.23%) of pure homogeneous nucleoside 10, which from little methanol crystallized. M.p. 224-226 ° C, 1 H NMR (DMSO-D₆) δ: 2.99 (tr, 2 H; CH₂-C = C), 3.74 (tr, 2 H; CH₂-N), 3.79-3.92 (m, 2 H; H₅ ′), 4.13 (m, 1 H, H₄ ′) 4.21 (m, 1H, H₃ ′), 4.28-4.32 (m, 1 H; H₂ ′), 7.48 (s, 1 H, H-C = C)

Example 4 3-β-D-ribofuranosyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one 11

16.15 g (27.8 mmol) of 2 ', 3', 5'-tri-O-benzoate 10 were 72 h in 250 ml of saturated methanoli Leave ammonia, after which the thin layer chromatogram in the n-butanol system: AcOH: H₂O = 4: 1: 5 (upper phase) no longer indicated starting material 10. At the Ab the free nucleoside 11 already began to crystallize. The residue was in 300 ml of H₂O and 200 ml of methyl t-butyl ether added and after separation of the phases aqueous phase again extracted with 200 ml of methyl t-butyl ether. After narrowing the aqueous phase to 150 ml, 2 g of activated carbon became the yellow, hot aqueous solution given, the filtered coal washed with 75 ml of hot H₂O and the combined H₂O filtrates evaporated to give 7.8 g of crude crystalline nucleoside. In the Recrystallization from a mixture of 175 ml of ethanol and 40 ml of H₂O was 4.8 g of 11  M.p. 224-226 °, obtained and from the mother liquor still 1.84 g 11. total yield 6.64 g (88.8%) 1 H NMR (D₂O) δ: 3.7-3.9 (m); 4.07-4.12 (n, H₄ ′) 4.17-4.22 (m, H₃ ′), 4.27-4.30 (m, H₂ ′), 5.92-5.95 (m, H₁ ′), 7.47 (s, H-C = C)

Example 5 2,2′-anhydro-3 (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidine 2-on hydrochloride 14

To a suspension of 1.35 g (5 mmol) of compound 11 in 50 ml abs. Acetonitrile was added the 3.29 g (20 mmol) acetoxyisobutyric acid chloride at 21 ° and the reaction mixture The mixture was stirred at 21 ° for 4 h, a clear, colorless solution being formed after about 1 h and after 4 h the starting material 11 in the system upper phase of n-butanol-acetic acid-H₂O (4: 1: 5) was no longer detectable. After evaporating and stirring three times colorless viscous residue, each with 100 ml of diethyl ether, became 75 ml of 1 molar added methanolic HCl and stirred at 21 ° for about 18 h, whereby colorless crystals precipitated out len with 40 ml abs. Methanol washed and dried, 1.14 g (79%) 14, 235 ° (dec.). After the filtrate had been evaporated, 0.16 g remained in the residue. ¹H- NMR (DMSO-D₆) δ: 3.03-3.07 (m, CH₂-C = C), 3.4-3.43 (m, CH₂OH), 3.82-3.86 (m, N- CH₂), 4.22 (s, H-4 ′), 4.48 (s, H-3 ′), 5.04 (m, 5-OH), 5.4 (d, H₂ ′), 6, 5 (d, H₁ ′), 7.96 (s, HC = C)

Example 6 3- (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d) pyrimidin-2-one 15a and hydrochloride 15a · HCl

a) A suspension of 2.69 g (10 mmol) of ribofuranoside 11 in 150 ml of acetonitrile was 9.875 g (60 mmol) of α-acetoxyisobutyric acid chloride were added and the mixture was stirred at 24 ° for 5 h. After evaporation of the acetonitrile, the residue was washed twice with 300 ml of diethyl ether each stirred for 15 minutes, during which a fine white precipitate settled. The ether phase was decanted, the precipitate with 25 ml conc. Ammonia and the resulting clear solution left for 48 h. After evaporation was twice with 30 ml of H₂O and evaporated 30 ml of methanol. After adding 50 ml of methanol, colorless precipitated Crystals that were filtered off = 0.27 g 15, mp. 230-234 ° (dec.). After constriction the mother liquor and the addition of 30 ml of methanol were able to 0.743 g 15 after 18 h be filtered off, mp. 231-235 ° (dec.). After the mother liquor had evaporated, the Residue stirred with 30 ml of pyridine and 15 ml of acetic anhydride at 22 ° for 18 h. The light yellow After evaporation and two co-distillation with 2 × 40 ml of toluene, the solution gave one Residue, shaken with 60 ml ice-cold saturated NaHCO₃ solution - 100 ml CH₂Cl₂ has been. The CH₂Cl₂ phase was dried with Na₂SO₄ and evaporated. After loosening in  Ethyl ester was filtered through a column of 100 g silica gel. After 750 ml of flow resulted the next 750 ml of eluate 1.9 g of pure 2 ', 3', 5'-tri-O-acetate, which with 50 ml of methanolic NH₃ was saponified overnight. After evaporation, the residue (1.8 g) crystallized out Methanol and in two portions gave a further 0.7 g 15a, mp. 232-235 °. Overall yield of 15a = 1.71 g (63.5%). MS (EI) m / z = 269 (M⁺), 251 (M-H₂O), 238, 208, 177, 166, 150, 138, 121, 110, 95, 82, 70, 55, 43. 1 H-NMR (DMSO) 2.82 (tr, J = 8 Hz, 2 H, CH₂-C) 3.53 (tr, J = 8 Hz, 2 H, CH₂-N) 3.59 (m, 2 H, C₅-CH₂) 3.61 (m, 1 H, H₄ ') 3.89 (m, 1 H, H₃ ′) 3.92 (m, 1 H, H₂ ′) 4.9 (tr, J = 5 Hz, 1 H, 5 ′ OH) 5.3 (d, J = 6 Hz, OH) 5, 33 (tr, J = 4 Hz, 1 H, OH) 6.02 (d, J = 4 Hz, 1 H, H 1 ′) 7.37 (d, 1 H, H-C = C) 7.76 (s, NH).

b) Starting from 4.03 g (15 mmol) of D-ribofuranoside 11 gave the analogous reaction (see Example 5) with 9.875 g (60 mmol) of α-acetoxyisobutyl chloride 3.5 g of crude 2,2′- Anhydroproduct 14, which was stirred with 10% aqueous ammonia at 22 ° for 3 days. To Evaporation and twice adding 30 ml of H₂O, the oily residue was in 120 ml of methanol are mixed with 30 ml of 2N NaOH and evaporated. After suspending in 100 ml of methanol and stirring for 15 minutes, the colorless precipitate was filtered off and washed with 30 ml of methanol washed. As much methanolic HCl was added to the filtrates until a pH of 3 was reached, with NaCl failing. After stirring for 30 minutes, the NaCl filtered off and washed with 30 ml of methanol. The filtrates were evaporated and with absolute ethanol extracted, 3.28 g of Ansa-aracytidine hydrochloride 15a-HCl, Mp = 212-216 ° (dec.), Which was washed with 25 ml of methanol. The abge steamed mother liquors were dissolved hot in 35 ml of methanol, with further cooling 0.23 g 15a · HCl, mp 212-216 ° (dec.) Were obtained. Total yield at 15 HCl = 3.51 g (76.53%), MS (CI) m / z = 270 (M + 1) 168, 138, 1 H-NMR (D₂O) δ: 2.95 (m, 2 H; CH₂-C = C), 3.6 (m, 2 H; H₅ '), 3.78 (m, 1 H, H₄'), 3.58 (m, 2 H; CH₂N), 3.92 (m , 1 H; H₃ ′), 4.04 (m, 1 H; H₂ ′), 5.49 (m, 1 H, H₁ ′), 7.72 (s, 1 H; H-C = C)

Example 7 3- (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2-H-pyrrolo [2,3-d] pyrimidin-2-one-5'-O- phosphate 15b

A suspension of 0.404 g (1.5 mmol) of 3- (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2H- pyrrolo- (2,3-d) pyrimidin-2-one 15 in 7.5 ml triethyl phosphate was stirred at -25 ° slowly for one hour using a syringe with 0.173 ml (1.88 m mol) Phosphorus oxychloride added and then stirred at about -20 ° for 1 hour and then stirred in an ice bath for 2 hours at + 2 °. After stirring in 25 ml of ice water was still Stirred for 2 hours at 22 ° and the triethyl phosphate by extraction with 6 × 25 ml CH₂Cl₂ extracted from the aqueous solution. The aqueous phase then slowly became one Solution of 1 ml of 1,2-butylene oxide in 180 ml of ethanol. After stirring for 5 hours  The precipitated colorless substance was filtered off at 22 ° and washed with 20 ml of ethanol, whereby 0.39 g of 5'-phosphate 15b were obtained. After evaporating the filtrate, dissolve in 5 ml H₂O after stirring in 75 ml of ethanol fell another 0.093 g of 5'-phosphate 15b out. Overall yield = 0.483 g = 92.35% on 15b. MS (FAB / m / z = 372 (M · HO³).

Example 8 Sodium salt of 3 (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3] pyrimidine- 2-one-5'-O-phosphate 15c

From the 5'-O-phosphate described in Example 9, 0.200 g (0.57 mmol) was suspended in 10 ml H₂O and the pH of the solution was brought to 7 with 1 N NaOH, a clear solution being formed. After stirring in 70 ml of ethanol, the mixture was stirred at 22 ° for 2 hours, filtered off and washed with 15 ml of ethanol, 0.196 g (92.9%) of pure monosodium salt of 5′-O-phosphate-15c- being obtained after drying in vacuo, MS (FAB) m / z = 394 (M + Na) 372 (M-HO³) 237, 133 IR / KBr) 1680, 1575, 1505, 1315 cm ^-1

Example 9 3- (2′-deoxy-3 ′, 5′-di-O-toluoyl-β-D-ribofuranosyl) -7-acetyl-3,5,6,7-tetrahydro-2H- pyrrolo [2,3-d] pyrimidin-2-one 19 and 3- (2′-deoxy-3 ′, 5′-di-O-toluoyl) -α-D- ribofuranosyI) -7-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimid-in-2-one 20

1.79 g (10 mmol) 7-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one 1 (R = COCH₃; R '= H) were with 2.1 ml (10 mmol) hexamethyldisilazane (HMDS) and 1.27 ml (10 mmol) trimethylchlorosilane (TCS) in 45 ml of acetonitrile boiled under reflux for 2 h, with a clear solution formed and NH₄Cl sublimed into the cooler. After evaporating the residue three times with 15 ml of abs. Xylene evaporated. The crystalline back was suspended in 120 ml of abs. 1,2-dichloroethane and added 3.86 g (10 mmol) of 3.5- Di-O-toluolyl-2-deoxy-α-D-ribofuranosyl chloride and then at + 5 ° 1.99 ml (11 mmol) trimethylsiltriflate in 40 ml abs. 1,2-dichloroethane for 30 min. with stirring added. After a further 15 min at + 7 °, the mixture was stirred for about 4 h at + 21 ° and the reaction mixture stored in the refrigerator at + 4 ° for 18 h. When working up with 120 ml ice-cold NaHCO₃ solution precipitated some red-brown substance, and an emulsion was formed. To Filtration over a G 4 frit and washing with CH₂Cl₂, the phases were separated and the aqueous phase extracted twice with 60 ml CH₂Cl₂. The combined organic phase after drying (Na₂SO₄) and evaporating, gave 4.75 g of a brownish amorphous crude product, dissolved in 40 ml of ethyl acetate - CH₂Cl₂ (1: 1) and on a column of 180 g of silica gel was chromatographed. Elution with 1 l of ethyl acetate - CH₂Cl₂ (1: 1) gave 0.15 g of red-brown Oil, while the next 0.5 l 3.4 g almost colorless, partially crystalline α / β mixture of 19  and 20 resulted. Elution with another liter of ethyl acetate: CH₂Cl₂ mixture gave 0.43 g almost pure α-nucleoside 20, which crystallized from a little acetone. Crystallization of the 3.4 g α / β- Mixture of about 100 ml acetone -CH₂Cl₂ gave 1.5 g (28.2%) of crystalline pure β-nucleoside 19, mp 203-205 °. The mother liquor contained according to the thin layer (upper phase of Toluene-acetic acid-H₂O = 5: 5: 1) mainly the α-anomer 20 and was therefore with inoculated crystalline α-anomer in a little acetone, with further crystalline α-anomer 20, 148-150 °.

β-anomer 19 1 H-NMR (CDCl₃) δ: 2.26-2.23 (m, 1 H, CH₂-N), 2.41 + 2.44 (s, 2 × CH₃-Ar), 2.40-2.50 (m, 1 H, CH₂-N), 2.60-2.70 (m, 1 H, H₂ ′), 2.69 (s, CH₃CO), 3.07 (d, d, d, J = 15 Hz; 1 H; H₂ ′) 3.83-3.90 + 3.94-4.02 (m, 2 × 1 H; N-CH₂), 4.60 (dd, J = 3 Hz, 11 Hz, 1 H, H₅ ′), 4.63 (dd; J = 2 Hz, J = 3 Hz; 1 H; H4 ′), 4.68 (dd, J = 2 Hz; J = 11 Hz; 1 H; H₅ ′), 5.62 (d, J = 7 Hz; 1 H; H₃ ′), 6.39 (dd; J = 6 Hz; J = 8 Hz; 1 H, H₁ ′), 7.24 (d, J = 8 Hz, Ar-H), 7.28 (d, J = 8Hz, Ar-H), 7.64 (s, 1H; C = C-H), 7.85 (d; J = 8Hz, ArH), 7.97 (d; J = 8Hz, ArH)

Example 10 3- (2'-deoxy-β-D-ribofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3d] pyrimidin-2-one-21

A suspension of 1.3 g (2.45 mmol) of 3- (2'-deoxy-3 ', 5'-di-O-toluoyl-β-D-ribofuranosyl) - 7-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3d] pyrimidin-2-one 19 in 150 ml methanolic Ammonia was stirred at 22 ° for 48 h, a yellowish, clear solution being formed. To The residue was evaporated in 150 ml of H₂O and 100 ml of methyl t-butyl ether taken. After the phases had been separated, the aqueous phase was evaporated off. The colorless, crystalline residue (0.68 g) was recrystallized from 5 ml H₂O, 0.22 pure 21, Mp 135-140 ° → 223-226 ° was obtained. Recrystallization of the evaporated Mother liquor from 8 ml ethanol-H₂O (9: 1) gave a further 0.17 g 19, mp. 134-140 ° → 224-228 °. Overall yield = 0.39 g 21 (63%).

Example 11 3- (2 O-acetyl-5-O-benzoyl-2-deoxy-2-fluoro-β-D-arabinofuranosyl) -N⁷-acetyl-3,5,6,7- tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one

The Ansacytoxin I (R = Ac: R ′ = H) 0.896 g (5 mmol) was in 40 ml abs. Acetonitrile with excess hexamethyldisilazane (HMDS) trimethylchlorosilane (TCS) 30 minutes boiled, forming a clear solution of the silylated compound 8b and NH₄Cl in sublimated the reflux condenser. After evaporating the solution in vacuo Exclusion of moisture, the residue was twice with 15 ml of abs. Xylene in Evaporated vacuum, in 20 ml 1.2 abs. Dichloroethane dissolved and a solution of 1.206 g (5mmol) of 3-0-acetyl-5-0-benzoyl-2-deoxy-2-fluoro-α-D-arabinofuranosyl bromide 22  added and the reaction mixture boiled under nitrogen for 14 hours. After cooling and diluting the solution with 50 ml of CH₂Cl₂ was shaken with 75 ml of ice-cold sat. NaHCO₃ solution and extracted the aqueous solution with 2 × 30 ml CH₂Cl₂. To Drying (Na₂SO₄) and evaporation, the 2.53 g of crude product in 20 ml of the upper phase Mixed toluene: acetic acid: water = 5: 1: 1 dissolved and on a column of 120 g Chromatographed silica gel. After a preliminary run of 500 ml the next 150 ml resulted 0.812 g (35.35%) of one β-anomer, while the next 150 ml 0.210 g β-α mixture and the last 150 ml gave 0.040 g (1.7%) of pure α-anomer.

Total nucleoside yield = 1.062 g (46.2%)
β-anomer: MS (EI) m / z = 459 (M⁺), 439 (M-1 + F) 380, 338, 318, 281, 258, 219, 180, 137, 105 MS (CI) = 460 ( M + H³)
1H-NMR (CDCl₃) 2.18 (s, 3H, COCH₃) 2.7 (s, 3H, N-COCH₃) 2.68 (m, 2H, CH₂CH₂N) 4.05 (tr, 2H, J = 8Hz, CH₂N) 4.4 (tr, 2H, H₄) 4.68 (dd, 2H, J = 4, 12Hz, H-5 ′) 4.76 (dd, 2H, J = 3 + 12Hz H-5 ′, 5.30 (dd, 1H, J = 2 + 17Hz, H'₂), 6.28 + 6.34 (dd, 1H; J = 2, 17Hz, H'₁) 7.2-8.08 (m, aroma. ) 7.58 (s, 1H, HC = C), 4.05 (tr. 2H; J = 8 Hz, CH₂N)

Example 12 3- (2-deoxy-2-fluoro-β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3- d] pyrimidin-2-one 23

0.77 g (0.168 m mol) of pure β-anomer 3- (2′-O-acetyl-5′-O-benzoyl-2′-deoxy-2′-fluoro-β-D- arabinofuranosyl) -N⁷-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo- [2,3-d] pyrimidin-2-one were 20 Stirred at 22 ° with 50 ml of methanolic ammonia and evaporated in vacuo. The residue was washed with 150 ml of water and 75 ml of methyl t-butyl ether (MBE) recorded and after separating the phases, the aqueous phase twice with 50 each ml of MBE and extracted with 75 ml of CH₂Cl. After evaporation of the aqueous phase, the Residue dissolved in 10 ml of water, mixed with 0.2 g of activated carbon and filtered off Wash the activated carbon with 10 ml of water. After evaporation of the aqueous phase was obtained 0.54 g of crude product, which after dissolving in 30 ml of boiling ethanol on cooling 0.290 g pure 23 melting point 219-221 ° and when concentrating the mother liquor to 10 ml more 0.038 g 23 together delivered.

MS (EI) m / z = 271 (MO³), 251 (M-H₂O), 204, 177, 137, 121, 110, 93, 82 MS (CI) m / 7 = 272 (M + 1) 252, 177.138.

1 H-NMR (DMSO) 285 (tr, 2H, J = 8Hz, CH₂CH₂N), 3.5-3.6 (m, 3H, CH₂N) 3.78 (m, 1H, H4 ′) 4.12-4.21 (m, 1H, H₃ ′), 4.83 (dd, J = 2 + 3Hz, H′₂), 6.08 + 6.13 (dd, J = 3 + 17 Hz, H'₁) 7.4 (s, 1H, H-C = C)  

Example 13 3- (3,5-di-0-benzoyl-2-deoxy-2,2-difluoro-β-D-ribofuranosyl) -N⁷-acetyl-3,5,6,7- tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one

A suspension of 0.896 g (5 m mol) 1 (R = Ac, R '= H) in 50 ml abs. Acetonitrile was with excess hexamethyldisilazane (HMDS) and trimethylchlorosilane (TCS) 90 Boiled minutes, the silylated compound 8b formed and NH₄Cl in the Reflux cooler sublimated. After evaporation, the residue (8b) was 3 to 4 hours 40-50 ° (10-² bar) dried and in 100 ml abs. 1,2-dichloroethane dissolved. 2.2 g (4.8 m mol) 3,5-di-0-benzoyl-2,2-difluoro-D-ribofuranosyl mesylate 25 to the solution and dripped for 15 minutes a solution of 0.93 ml (5 m mol) trimethylsilyl triflate in 20 ml abs. 1,2-dichloroethane at 22 ° with stirring. The reaction mixture was then 14 Boiled for hours at reflux, a further 0.37 ml (2 mol) of trimethylsilyl triflate in 20 ml of abs. 1,2-dichloroethane at 22 ° with stirring. The reaction mixture was then 19th Boiled for hours at reflux, a further 0.37 ml (2 m mol) of trimethylsilyl triflate added and cooked again for 5 hours, after which only traces of the starting base in the DC. system (Upper phase of toluene: acetic acid: H₂O = 5: 5: 1) were visible. After working up with CH₂Cl₂, ice-cold NaHCO₃ solution, the CH₂Cl₂ phase dried (Na₂SO₄), filtered and evaporated. The 2.64 g reddish brown, oily raw product were then with the upper phase of toluene: acetic acid: H₂O = 5: 5: 1 on a column of Chromatographed 120 g of silica gel. After approx. 550 ml lead, the next 550 ml resulted Eluate 0.73 g mixture while the next 550 ml 0.48 g pure protected α-anomer delivered. A second chromatography of the 0.73 g mixture again on 120 g silica gel After approx. 650 ml lead, delivered 0.540 g pure β-anomer and 0.050 g pure α-anomer.

β-anomer: MS (CI) m / z = 540 (M⁺ + H); 498; 478; 436
1 H-NMR (CDCl₃) δ = 2.48 (tr; 2H: j = 8Hz, CH₂CH₂N) 2.53 (s, 3H; CH₃CO) 4.3 (tr; 2H; J = 8Hz, CH₂N) 4.6 (m, 1H, H₄ ′), 4.68-4.9 (m, 2H, H₅ ′), 5.63-5.68 (m, 1H, H₃ ′) 6.58-6.63 (m, 1H, H′₁) 7.41 (s, 1H, HC = C), 7.45- 8.13 (m, aroma H).

Example 14 3- (2-deoxy-2,2-difluoro-β-D-ribofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] - pyrimidin-2-one 26

A suspension of 0.480 g (0.89 mol) of 3- [3,5-di-O-benzoyl-2-deoxy-2,2-difluoro-β-D-ribofuranosyl) N-7-acetyl-3,5 , 6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimid-in-2one in 40 ml of methanolic ammonia was stirred at 22 for 18 hours, a clear solution being formed after about 30 minutes. After evaporation and work-up with water -CH₂Cl₂ and subsequent extraction of the aqueous phase with CH₂Cl₂ and ethyl acetate, the aqueous phase was concentrated and the residue (0.26 g) was recrystallized from 5 ml of water, 0.161 g (62.5% ) pure 26, melting point 167-170 ° were obtained. MS (CI) m / z = 290 (M + H) 138, MS (EI) m / z = 290, 289 (M⁺) 272, 215, 208, 200, 95, 172, 164, 138, 137, 121, 110, 109, 93, 82, 60, 44
1 H-NMR (DMSO-D₆) δ = 3.58 (tr, 3H, J = 8Hz, CH₂CH₂N), 3.63-3.68 (m, 1H, H₄ ′), 3.76 (m, 1H, H₅ ′) 4.13- 4.18 (m, 1H, H₃ ′) 6.1-6.15 (m, 1H, H₁ ′), 7.43 (s, 1H, HC = C).

Claims (4)

1. Cytosine and cytidine derivatives of the general formula I wherein
R is hydrogen, CHO, COR⁶ with R⁶ in the meaning of C₁-C₁₇-alkyl and phenyl, or COOR⁷ with R⁷ in the meaning of C₁-C₄-alkyl or benzyl
R ′ is hydrogen or the groups wherein
n 1 or 2
X 0, S, CH₂ or NR,
R² is hydrogen, fluorine, CH₃ or CN,
R³ is hydrogen, fluorine, hydroxy or -OCH₃,
R⁴ is hydrogen, fluorine, hydroxy, N₃ or NH₂,
R⁵ is hydrogen or PO (OH) ₂,
R², R³ together = CH₂,
R³, R⁴ together form a double bond and their salts. 2. 3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one
7-acetyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one
3-β-D-ribofuranosyl-3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one
3- (β-D-arabiflofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d) pyrimidin-2-one
3- (β-D-arabinofuranosyl) -3,5,6,7-tetrahydro-2-H-pyrrolo [2,3-d] pyrimidine - 2-one-5'-O-phosphate
3- (2'-Deoxy-β-D-ribofuranosyl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one
3- (2-Deoxy-2-fluoro-β-D-arabinofurano syl) -3,5,6,7-tetrahydro-2H-pyrrolo [2,3-d] pyrimidin-2-one. 3. Use of the compounds according to claim 1 and 2 for the preparation of Medicines. 4. Process for the preparation of the compounds of formula I. wherein the radicals R 'and R have the meaning given above, characterized in that

• a) a compound of formula 7 with triphenylphosphine dihalide and a tertiary amine or
• b) a formylpyrrolidone derivative of the formula 17 in which R¹³ is hydrogen, an SiR¹⁴R¹⁵R¹⁶ radical in which R¹⁴, R¹⁵, R¹⁶ are identical or different and are phenyl or C₁-C₄-alkyl, or is tert-butyloxycarbonyl,
  with a urea derivative or an isourea derivative such as 16,
  wherein R¹⁷, R¹⁸ are the same or different and are hydrogen or SiR¹⁴R¹⁵R¹⁶, reacted and, if desired, subsequently formylated, acylated or COOR⁷ or
  in the 3-position introduces the optionally hydroxyl-containing radical R 'and then, if desired, introduces N₃, NH₂, fluorine, CN or = CH₂ or R³, R⁴ in the meaning of a double bond and if desired hydrogenates or 5' hydroxyl groups in the 5'-phosphate transferred and, if necessary, splitting off protective groups or forming the salts.