Di(2-PROPYL)ESTERS OF 1-FLUORO-2-PHOSPHONOMETHOXY-3-P-TOLUENESULFONYLOXYPROPANES, THEIR PRODUCING AND UTILIZATION

Abstract: The invention relates to new compounds, di(2-propyl)esters of (R)-, (S)- and (RS)-1-fluoro-2-phosphonomethoxy-3-p-toluenesulfonyloxypropane and the method of producing them. The compounds can be used with advantage in the preparation of N-(3-fluoro-2-phosphonomethoxypropyl) derivatives of heterocyclic purine and pyrimidine bases that have antiviral activity.

Description: P (2-propyl )esters of l-fl.uρrp..-2-p,hp£Phpnpfflethpχ -~7--r--t.ρ- luenesulfonylo,:vpropanes, their producing and utilization Technical Field This invention relates to di(2-prop 1 ) esters o 1-fluoro- 2-phosphonomethoκy-3-p-tolLienesulfonylctypropanes, the method of their preparation and utilization in production of antivirals, N-(3-fluoro-2-phosphonometho,\ypropy1 ) derivatives of heterocy lic purine and pyrimidme bases. Bachqrpunti Art N-<3-Fluoro-2-phosphonometho ϊyprop 1 ) derivatives of hete¬ rocyclic purine and pyrimidme bases <FPI P-deπ ati eε) occupy a significant place among compounds effective against retro- viruses causing serious illnesses in man and animals. Under in vitro conditions, these compounds exhibit biological parameters (selectivity of action better than e.g. c ?-(2-phosphonometho>:yeth 1 )adenιne (R.Pau els, J. Balzarin , D. Schols, l . Baba, J. Deεmyter, I. Rosenberg, A. Holy-, E.De Clercqi Antimicrob. Ag. Chemother. I2, 1025 (1^88^. The said derivatives were hitherto accessible from N-(3-fluoro- 2-hydro*?ypropy11 derivatives of purine or pyrimidme bases (prepared e.g. according to CS-patent PV 2048-^0> by reaction with diallyl p-toluenesulfon lo 'y- methanesul fonates or ethanesLilfonylccymethanephαsphonateε in the presence of sodium hydride and subsequent removal of the protecting groups by acid or alkaline hydrolysis and f lli with bromotrimethylsilane. The drawbacl of this method (according to CS-patent application PV 2047- t ?Cπ is that it is necessary first to prepare the optically act! -J& N-f3-fluoro-2-hydro;;ypropyl "* derivatives and perform their multistage protection. Another possible solution would be condensation of the heterocyclic base (its silyl derivative or alkali metal salt) with a suitable optically active organophosphorus synthon that has a preformed structure of the side chain of FPMP derivatives. Such a reaction of suitably protected derivatives of glycerol with dial yl esters of p-toluenesLilfonyloκymethanephoεphonic acid, followed by partial deblocking and introduction of the reactive tosyl or esyl group has been utilized e.g. in producing analogous N-(3-hydroκy-2-phoεphonometho>;ypropyl) derivatives (J. J. Bronson, I. Ghazzouli, M. J. M. Hitchcock, R. R. Webb, J. C. Martin: J. Med. Chem. 32, 1457 (198?)). Still another approach made use of introduction of the esterified phosphonomethyl ether functionality on the hydrσxyl component via an acetoκymethy1 ether by treatment with bromotrimethylsilane and trialkyl phosphite (CS-patent application PV 3871-90). Disclosure pf Invention The above-mentioned drawbacks in the so far Lised methods of preparing N-(3—fluoro-2-phosphonomethoκypropy1) derivatives of PLtrine and pyrimidine bases are removed by this invention relating to di(2-ρropyl) esters of i-fluoro-2-phoερhono- methoκy-3-p-toluenesLtlfonyloMypropanes of the general formula I ChfeC 6 H 4 S0 2 0CH 2 ( I) where the absolute configuration at carbon atom C-2 is R, S or RS, and the method of preparing them which is characterized in that i-flLioro-2,3-propanediols of the general formula II FCHaCHCH 2 OH (II) I OH where the absolute configuration at carbon atom C2 is R, S or RS, are reacted with p-toluenesulfony1 chloride in the presence of a tertiary amine, preferably triethylamine or pyridine, in an inert organic aprotic solvent, preferably benzene or dichloromethane, or only in pyridine, the obtained 1-flu.oro-3-p-toluenesulfonyloxy-2-propanolε of the general formula III FCH2CHCH2OSO2C6H4CH3 (III) OH where the absolute con iguration at carbon atom C2 is R, S or RS, βire reacted with dimethoxymethane in the presence of phosphorus pentoxide in an inert organic solvent, preferably dichloromethane, the resulting 1-fluoro—2—methoxy ethoxy-3-p-toluenesulfonyloxy- propanes of the general formula IV CH 3 C 6 H 4 SO2OCH2 ( I ) where the absolute configuration at carbon atom C2 is R, S or RS, are treated with acetic anhydride in the presence of a Lewis acid, preferably boron trifluoride etherate, at -5 to +5 °C, the obtained 2-acetoκy ethoxy-l-fluoro-3-p-toluenesulfony1- oxypropanes of the formula V OCH2 where the absolute configuration at carbon atom C2 is R, S or RS, are heated with bromotrimethylsilane to 100 - 120 °C, preferably in toluene, the volatile material is evaporated in vacuo and the residLie is heated with tri(2-propyl ) phosphite at 100 - 120 °C, and the compounds of the general formula I are isolated, preferably by chromatography. The invention also relates to the utilization of the said compounds of the formula I in the preparation of N-(3—fluoro—2-phosphonomethoxyprop 1 ) derivatives of hetero- cyclic purine and pyrimidine bases of the general formula VI B-CH2CH-OCH2P ( 0 ) ( OH ) ( VI I CH 2 F where the absolute con guration at carbon atom C2 is R, S or RS and B is s. su.b__titu.ted ρuriπ-?-; * I 7 purin-7-yl, pyrxmidi'-.-i-yl or pyrimidine-3-yl moietj or their ars t_ diaza analogues, which is characterized in that compounds of the formula τ . Ξ.ΓΞ reacted with a sodium salt of the substituted purine or pyrimidine heterocyclic base, its aza or diaza analogue, or their acyl, preferably benzoyl, derivatives, or with a mixture of these bases with an alkali metal carbonate, preferably cesium carbonate, in di ethylformamide at 60 - 140 °C, whereupon the solvent is evaporated in vacuo and the residue is treated with sodium ethoxide in ethanol and then with bromotrimethylsilane in an inert organic solvent, preferably in acetonitrάle and the resulting compounds of the general formula VI are isolated by chromatography, preferabl on ion-exchanging resins. Devising an organophosphon-is synthon suitable for the preparation of FPMP-derivatives of the general formula VI involves the following problems: (a) the choice of a su.itable reactive group capable of substituting the active form of the heterocyclic base, (b) the choice of suitable ester protecting groups on the phosphonic acid moiety, and (c) carrying out the whole synthetic sequence leading to the chiral synthon from an easily accessible chiral compound with preservation of optical purity of both the synthon and the final product. The starting fluoro derivatives of the formula II are prepared e.g. by reaction of 1-0-p-toluenesulfonyIglycerols with alkali metal fluorides; this reaction can be realized with the racemic compound (CS-patent application PV 2048-90) as well as with both the enantio eric derivatives. In such case one can make use of p-toluenesulfony1 esters of 4-hydroxymethyl-2,2-dimethyl- 1,3-dioxolanes of the formula VII of which the (R)-enantiomer is readily accessible from D-mannitol (A. Holy: Collect. Czech. Chem. Commun. 4_0_, 187 (1975)) whereas the (S)-antipode can be obtained e.g. from D~ribonolactone. Thus, e.g. a solution of 2,3-D-i5oprop li- dene-5-0-( -toluenesulfon 1 )-D-ribonolactone (50 g) (L. Hough, J. K. N. Jones, D. L. Mitchel: Can. J. Chem. _2ώ, 1720 (1958)) in dioxane (500 ml) is stirred with concentrated hydrochloric acid (50 ml) at room temperature for 12 h. Then water (130 ml) is added and the mixture is neutralized by addition of solid sodium hydrogen carbonate. The bottom aqueous layer is separated, washed with ethyl acetate (500 ml), and the combined ethyl acetate and dioxane solutions are dried over magnesium sulfate. After filtration, the solvents are evaporated and the dry residue is crystallized by addition of toluene (500 ml) to its boiling solution in ethyl acetate (130 ml); yield 31.6 g (71%) of 5-0-( -tolueneεulfon- y1 )-D-ribonolactone. A solution of this compound (30.7 g) in a mixture of water (75 ml) and dioxane (75 ml) is cooled to 0 °C and a solution of sodium periodate (22.8 g) in water (165 ml) is added. The mixture is stirred at room temperature for 20 miπ, cooled to 0 °C, and after 20 min dioxane (150 ml) is added. After filtration, sodium borohydride (4.73 g) is added in portions to the filtrate, the temper tire being kept below 20 °C. After 0.5 h at room temperature, the reaction mixture is poured into ethyl acetate (1.3 liter) and shaken SLiccessively with 1M HC1 (saturated with sodium chloride) (2 x 150 ml) and a 1:1 ixttire of saturated solutions of sodiu.m chloride and sodium hydrogen carbonate (2 x 150 ml). The ethyl acetate soluition is then dried over magnesium sulfate, filtered and the solvent is evaporated to dryness, yielding 20 g (80%) of (S)-3-0-(p-toluenesulfony1 ) lycerol. A solution of this compound (27 g) in acetone (150 ml) is stirred with 2,2—dimethoxypropane (20.2 ml) and trifluσromethanesulfonic acid (10 j l) at room temperature. After standing overnight at ambient temperature, triethyla ine (0.5 ml) is added, the reaction mixture is concentrated to a minimum volume, the residue is dissolved in toluene (500 ml), washed with water (3 x 250 ml) and dried over magnesium sulfate. The drying agent is filtered off and the filtrate is evaporated in vacuio to afford (S)-l,2-0-isopropylidene-3-0-(p-toluenesulfony1 )- glycerol (28.3 g). Reaction of 1,2-isopropylidene-3-0-(p-toluenesulfony1 )- glycerols with potassium fluoride in diethylene glycol affords flLioromethyl derivatives which can be converted into the starting compounds of the formula II (W. J. Lloyd, R. Harrison: Carbohyd. Res. 2-_>, 133 (1971)). Thus, e.g., a mixture of (S)-l,2-0-isopropylidene-3-0-(p-toluenesulfon- yl)glycerol (100 g), potassium flLioride (30 g) and diethylene glycol is heated to 160 - 170 °C for 2 h Linder vigorous stirring and introduction of nitrogen. The product which distills is collected in a receiver cooled with dry ice; yield 21 g (45%) of (S)-l-deoxy—1-fluoro-2,3-0-isopropylide- neglycerol. A mixture of this produ.ct and 1M HC1 (50 ml) is stirred at room temperature for 10 h. Then it is neutralized with an anion-exchanging resin (HC03~ form), filtered, ana the resin is washed with methanol (50 ml). The combined filtrates are treated with charcoal and after filtration the solvent is evaporated and the product distilled in a vacuum of an oil pump at bath temperature not exc eding 140 °C; yield 12.4 g (84%) of (S)-3-fluoro-1,2-propanediol . As follows from the previous studies, for the desired N-substitution of heterocyclic bases the presence of a reactive chloro, p-toluenesulfonyloxy or methanesulfonyloxy group in the synthon I represents an advantage. p-Toluenesulfony1 derivatives are well accessible from alcohols and can be easily purified. Therefore, the synthon I can be prepared preferably from 1-fluoro-3-p-toluenesulfon- yloxy-2-propanol of the formula III, which can be easily- obtained from compounds II by tosylation in pyridine or in inert organic solvents (benzene, chlorinated hydrocarbons) in the presence of a slight excess of a tertiary base (e.g. pyridine, triethylamine, di(2-propy1 )ethylamine) . The tosyl derivatives III are purified by chromatography on silica gel. The introduction of the phosphonomethyl ether functionali¬ ty on the carbon atom in position 2 in compound III makes use of 2-methoxymethyl ether IV as the key intermediate. This is easily prepared from compound III by acid-catalyzed transaceta-lization with dimethoxymethane in the presence of phosphorus pentoxide. The reaction is heterogeneous but the product is obtained in high yield by chromatography of the organic phase. The methoxy ethy1 group in compound IV can be converted into the acetoxymethy1 group by reaction with acetic anhydride in the presence of a Lewis acid, e.g., boron trifluoride etherate, and the obtained acetoxymethoxy derivative V is immediately converted into the reactive 2-bromomethyl ether by treatment with bromotrimethylsilane. This reaction proceeds sluggishly even at relatively high temperatures and it is advisable to follow its course by thin-layer chromatography. The solvent and acetσxytri ethy1- siiane are evaporated in vacuo and the crude product is immediately reacted with trialkyl phosphite. Tri(2-propyl )phosphite appears to be reagent of choice: it is sufficiently reactive, the 2-bromopropane arising in the reaction is volatile and can be removed during the reaction, and the formed di(2-ρropyl) ester of phosphonic acid in the synthon I is not capable of alkylatiαπ of the heterocyclic base, unlike e.g. the corresponding ethyl, or particularly ethyl, ester. However, synthons analogous to the compound I, containing other ester groups such as dimethyl, diethyl or di(2,2-dimethy1propy1 ) ester etc. can be prepared by the analogotis reaction with the corresponding trialkyl phosphite. This reaction proceeds smoothly and after its end and evaporation of volatile products the desired product, synthon I, is obtained by chromatography on silica gel. All stages of this reaction sequence are realized in high yields so that the final overall yield from the starting compound II exceeds 40%. The reaction steps require no special materials or equipment and the optical purity of the end product I is determined only by the optical purity of the compound II. It follows from the optical purity of the known compounds of the formula IV that are ultimately prepared from the synthon I by condensation with sodiLim salts of heterocyclic bases. Such reaction takes place in dimeth lformamide with a suspension of the soditim salt obtained e.g. by in situ reaction of the base with an equimolar amount of sodium hydride. An equimolar amount of compound I is used in the reaction which is carried out at 80-100 °C under exclusion of air moiεttire. Another particularly advantageous modification of this method consists in heating compound I with the heterocyclic base in the presence of cesium carbonate which proceeds substantially faster and under formation of less side-products. With purine derivatives of the formula VI, which are of particular interest becaise of their antiretroviral activity, such alkylation with sodium salts of some bases (e.g. adenine, 2, -diaminopu.rine, 6-methylthiopurine, 3-deazaadenine ': takes place invariably in the N9-position, with other bases (hypoxanthine) in position N7? with some other bases (e.g.- guanine, 2-aminopurine, 1-deazaadenine) a mixture of the N9- and N7-isomers is formed. In such cases it is advantageous to use indirect methods based on subsequent conversion of the heterocyclic base in compounds of the general formula VI or their precursors modified at the base. Such is the case e.g. in the preparation of the hypoxanthine or xanthine derivative by deamination of compound, containing adenine or guanine, with nitrous acid or its esters, or in the synthesis of the 2-hydroxyadenine derivative by specific deamination of compound derived from 2,6-diaminopurine. The guanin-9-yl derivative as well as its N7-iεomer are prepared by hydrolysis of compound obtained from 2-amino-6-chloropurine. The same intermediate can be used to prepare 2-amino-6-substituted derivatives. Thus, compound of the formula VI containing 2,6-diaminopurine is obtained by hydrogenation of the corresponding 2-amino-6-azidopurine compound. The reaction of compounds I can also be performed with pyrimidine? derivatives; in such alkylations it is most important to ensure the regioεpeci ic course of the reaction in position Mis this is preferably achieved using 4~alkoxy-2-pyrimidoneε. The obtained Nl-substituted derivatives may be then converted by an acid hydrolysis with aqueous mineral acid or e.g. dilute acetic acid uracil or thy ine derivatives or, alternatively, by a inolysis, e.g. with alcoholic solutions of ammonia, primary or secondary amines at elevated temperatures to the derivatives of cytosine, 5-methylcytosine or their N-substituted analogs. The intermediate obtained by condensation of compound I with the base is either isolated by chromatography and is further processed as described below or the reaction mixture is directly methanolyzed (which removes the N-acyl group from the heterocyclic base). In the latter case, the obtained mixture is deionized, e.g. on a column of cation-exchanging resin (such as Dowex 50) in H + -form, and the di(2-prop 1 )ester of compound VI is isolated; this compound is sufficiently stable to withstand e.g. elution with dilute (Is10) aqueous ammonia from the resin. After drying, the ester groups in this intermediate are removed by reaction with bromotrimethylsilane, e.g. in acetoπitrile. After evaporation of the solvent, hydrolysis of the residue and desalting, the pure product of the formula VI is isolated by chromatography on an ion-exchanger or chromatography on hydrophobized silica gel. The optical purity of the thus-prepared compounds of the for Lila VI is proved e.g. by HPLC in 4 mM copρer(II) sulfate and 4 mM L— henylalanine at PH 3.1. The above-described procedure ffords compounds VI of optical purity >95%. The invention is fLirther illustrated by examples of execution without being in any way limited. Examples ,of .Execution Example 1. A solution of p-toltienesulfonyl chloride (26.4 g, 138 mol) in pyridine (30 ml) is added at -30°C to a solution of (S)-i-fluoro-2,3-propanediol (12.4 g, 132 mmol) in pyridine (30 ml). After standing at -30θC for 48 h, the reaction mixture is poured into ethyl acetate (1300 ml) and under ice-cooling shaken with 1M HC1 to acid reaction of the aqueoLiε layer, then with water, sodium hydrogen carbonate solution and again with water. After drying over magnesium sulfate and filtration, the solvent is evaporated in vacuo and the residue is chromatographed on silica gel (500 g) in toluene and then in a 50:1 mixture of toluene and ethyl acetate. Yield 21 g (64%) of (S)-l-fluoro-3-(p-toluenesulfon- loxy)-2—propanol. For Cι H ± 3 0*S (248.3) calculated: 48.37% C, 5.27% H, 7.65% F, 12.91% S; found: 43.54% C, 5.31% H, 7.43% F, 12.80% S. Phosphorus pentoxide is added at room temper ttire to a stirred mixture of this compound (11 g, 44.3 mmol), dimethoxymethane (11 ml) and dichloromethane (11 ml) uintil the starting compound disappears to afford the desired product (monitored by TLC in toluene-ethyl acetate 5:1). The mixture is filtered with Celite (10 g), the filter is washed with chloroform and the solvents are evaporated in vacuo; yield 11.9 g (93%) of (S)-i-fluoro~2-methσxymethoxy-3-(p-to- luenesulfonyloxy)propane. For Ci2H 1E :F0sS (290.3) calculated: 49.65% C, 5.20% H, 6.54% F, 11.04% S; found: 49.85% C, 5.34% H, 6.40% F, 10.91% S. A mixture of this product (2.45 g), acetic anhydride (5 ml) and boron trifluoride etherate (1 ml) is stirred at 0 °C for 2 h. Then the reaction mixture is shaken with a mixture of sodium hydrogen carbonate (10 g), water (100 ml) and toluene (100 ml). The aqueous layer is separated, the toluene layer is washed with water (2 x 50 ml) and dried over magnesium sulfate. Evaporation of the solvent gives (S)-2-acetoxymethoxy-l~fluoro-3-<p-toluenesulfonyloxy)propane in practically quantitative yield. For Cir-iHiTFO&S (320.1) calculated: 48.73% C, 5.35% H, 5.93% F, 10.00% S; found: 48.82% C, 5.51% H, 5.80% F, 9.95%S. This compound (11.0 g) is codistilled with toluene (3 x 10 ml), then toluene (20 ml) and bromotrimethylsilane (6.82 ml) are added and the mixture is heated to 80 °C for 16 h. Toluene is distilled off, the residue is codistilled with toluene (3 x 10 ml) and heated with triiεopropyl phosphite (10 ml) at 50 °C for 2 h. The mixture is again codistilled with toluene and the remaining oil is chromatographed on silica gel (100 g) by successive elutioπ with toluene, toluene-ethyl acetate (5:1) and toluene-ethyl acetate (1:1). Yield 11 g of (S)-i-fluoro-2-(diiεopropyIphos- phonyl ethoxy)-3-(p-tol ienesulfonyloxy)- propane; CKUD +12.50 (c 0.8, CHCI3). For CIT^SFOTPS (426.4) calculated: 47.87% C, 6.61% H, 4.45% F, 7.26% P, 7.51% S; calculated: 48.04% C, 6.97% H, 4.56% F, 7.45% P. MS, m/e (rel.%): 427 (M + H, 55), 385 (M - iPr, 17), 342 (M - 2 iPr, 100), 259 (9), 171 (TεO, 36). *H NMR (DMS0) : 1.20 d, 6 H (CH 3 , iPr) ? 1.26 d, 6 H (CH 3 , iPr); 2.43 ε, 3 H (CH 3 , Tε) ; 3.8 - 4.8 , 9 H (HC0, H 2 CF); 7.3 - 7.8 m, 4 H (arom. protons, Ts). Example 2 Phosphorus pentoxide is added to a stirred mixture of (R)-i-fluoro-3-( -toluenesulfonyloxy)-2-propanol (li g, 44.3 mmol), dimethoxymethane (11 ml) and dichloromethane (11 ml) at room temperature until the starting compound disappears in favour of the desired product (monitored by TLC in toluene-ethyl acetate 5:1). Celite (10 g) is added to the reaction mixture which is then filtered and the Celite is washed with chloroform. The solvent is evaporated from the filtrate to afford li.9 g (93%) of (R)-l-fluoro-2-methoxy- methoxy-3-(p-toluenesulfonyloxy)propane. This prodtict is processed as described in Example 1, yielding (R)-2-acetoxy- methoxy-3-fluoro-l-p-toluenesulfonyloxypropane in quantitative yield. For Ci3Hi F0 6 S (320.1) calculated: 48.73% C, 5.35% H, 5.93% F, 10.00% S; found: 48.82% C, 5.51% H, 5.80% F, 9.95% S. The product is processed according to Example 1, giving (R)—i-fluoro—2-(diisopropylphosphonylmethoxy)-3-(p-toluene- sulfoπyloxy>propane in 75% yield. EK3D -i2.6° (c 0.8, CHC1 3 ). For Ci7H2sF0 7 PS (426.4) calculated: 47.87% C, 6.61% H, 4.45% F, 7.51% S; found: 48.13% C, 6.91% H, 4.52% F, 7.35% S. MS, m/e (rel. %): 427 (M + H, 55), 385 (M - iPr, 17), 342 (M - 2 iPr, 100), 259, 171 (TεO, 36). H NMR (DMS0): 1.20 d, 6 H (iPr); 1.26 d, 6 H (iPr); 2.43 ε, 3 H (CH 3 , Ts) 5 3.8 - 4.8 m, 9 H (HC0, H2CF), 7.3 - 7.8 m, 4 H (arom. proton, Ts). Example 3 Sodium hydride (60% suspension in paraffin oil, 1.125 g, 28 mmol) is added to a suspension of N^-beπzoyladenine (6.5 g, 27 mmol in dimethylformamide (100 ml) and the mixture is stirred and heated at 60 °C for 15 miπ. After addition of (S)-l-fluoro-2-(diisopropylphosphonylmethoxy)-3-(p-toluene- SLilfonyloxy)propane (10 g, 23.6 mmol) the mixture is further stirred and heated at 80 °C for 6 h. The reaction mixture is then neuitralized with acetic acid and dimethylformamide is evaporated at 40 °C 100 Pa. The residue is extracted with chloroform, the chloroform extracts are filtered through Celite and the filtrate is concentrated in vacLio. The εiru.ρy reεidue is chromatographed on a column of silica gel (500 g) in a mixture of chloroform and methanol (50:1); yield 3.7 g (32%) of (S)-9-(3-fluoro-2-diisoρropylphosphonylmethoxy- propyl )-W 6 -benzoyladenine. For C22H29FWEO5P (493.5) calculated: 53.54% C, 5.92% H, 3.85% F, 14.19% N, 6.27% P; found: 53.71% C, 5.83% H, 3.71% F, 13.97% N. Sodium hydride (50 g, 2 mmol) iε added to a solution of this compound (2.1 g, 4.25 mmol) in methanol (20 ml). After standing at room temperature for 16 h, acetic acid (250 JJ1 > is added, the mixture is concentrated to a small volume and the residue iε chromatographed on a column of silica gel in a mixture of chloroform and methanol (30:1). Yield 1.11 g (67%) of (S)-9-C3-fluoro-2-(diisopropylphosphonylmethoxy)- propylϋadenine, m.p. 72-74 °C (toluene). For CisH2EFNEO.P (389.4) calculated: 46.26% C, 6.47% H, 4.87% F, 17.98% N, 7.95% P; found:47.26% C, 7.31% H, 4.90% F, 17.13% N, 7.26% P. MS, m/e (rel.%): 389 (M, 10), 374 (M - CH3 , 8), 288 (20), 209 (M - CH2P(0)(0iPr) 2 , 48), 195 (0CH 2 P(0) (OiPr) 2 , 100), 174 (12), 149 (AdeCH 2 , 40), 135 (Ade, 60). *H NMR (DMS0): 1.08 - 1.20 m, 12 H (CH3 , iPr) ; 3.7 - 4.7 m, 9 H (HC0, H 2 CF); 7.19 bs, 2 H (NH 2 ); S.09 s, 1 H (H-S, Ade); 8.15 s, 1 H (H-2, Ade). Bromotrimethylsilane (2.17 ml, 16.4 mmol) is added at room temperature to a solution of the above product (1.6 g, 4.1 mmol) in acetonitrile (8 ml) and the mixture is stirred at ambient temperature for 24 h. The reaction mixture is concentrated to a minimum volume, the residue is codistilled with toluene (2 x 10 ml) and with methanol (10 ml) and 2% aqueous solution of ammonia (10 ml) iε added. After standing for 1 h at room temperature, the solution is evaporated to dryness, the residue is dissolved in a minimum amount of water and the solution , is applied onto a column of cation-exchanger Dowex 50 (H+-form, 50 ml). The column is washed with water until the eluate exhibits negative reaction to bromides and then the product is eluted from the column with 2% aqueous ammonia. The obtained eluate iε concentrated to a minimum volume and applied onto a column of anion-exchanger Dowex 1 (acetate form, 50 ml). The column is washed with water to negative reaction to ammonia and the product iε eluted with 1M acetic acid. Evaporation of the solvents and codistillation of the residue with water affords 1.1 g (91%) of (S)-9-(3-fluoro-2-phosphonomethoxypropyl)- adenine, m.P.194-197 OC, Cκ3 D -16.9 (c 0.38, water). For C<?Hi3FNs0fl.P (305.2) calculated: 35.41% C, 4.29% H, 6.22% F, 22.94% N, 10.14% P; found: 35.20% C, 4.43% H, 5.85% F, 22.73% N, 10.44% P. Example 4 Sodium hydride (60% suspension in paraffin oil, 1.125 g, 28.1 mmol) iε added to a suspension of N 6 -benzoyladenine (6.5 g, 26.9 mmol) in dimethylformamide (100 ml) and the stirred mixture iε heated at 60 °C for 15 in. After addition of (R)-l-fluoro-2—(diisopropylphosphonylmethoxy)-3-(p-tolue¬ nesulfonyloxy)propane (10 g, 23.6 mmol), the mixture is further stirred and heated at 80 °C for 6 h. It iε then neutralized with acetic acid and the dimethylformamide iε evaporated at 40 °C and 100 Fa. The residue iε extracted with chloroform, the combined chloroform extracts are filtered through Celite and the solvent iε evaporated in vacuo. The sirupy residue is chro atographed on a column of silica gel (500 g) in a mixture of chloroform and methanol (50:1), yielding 3.7 g (32%) of (R)-)-(3-fluoro-2-(diisopropyl- phoεphonylmethoxypropyl) )-N 6 -benzoyladenine. For C22H ?F E0EP (493.5) calculated: 53.54% C, 5.92% H, 3.85% F, 14.19% , 6.27% P; found: 53.67% C, 5.99% H, 3.70% F, 13.95% , 6.40% P. Sodium hydride (50 mg, 2 mmol) is added to a solution of this product (2.1 g, 4.25 mmol) in methanol (20 ml). After standing for 16 h at room temperature, acetic acid (250 JJI) is added, the mixture is concentrated in vacuo to a minimum volume and chromatographed on a column of silica gel in a mixture of chloroform and methanol (30:1), affording 1.1 g (67%) of (R)-9-C3-fluoro-2-(diisopropylphosphonylmethoxy- prop l)3adenine, . p. 72-74 °C (toluene). For (389.4) calculated: 46.26% C, 6.47% H, 4.87% F, 17.89% N, 7.95% P; found: 47.16% C, 7.12% H, 4.81% F, 17.23% N, 7.46% P. MS, m/e (rel.%): 389 (M, 10), 374 (M - CH 3 , 8), 288 (20), 209 (M - CH 2 P(0) (OiPr) 2 , 48), 195 (0CH 2 P(0) (OiPr> 2 , 100), 174 (12), 149 (AdeCH 2 , 40), 135 (Ade, 60). *H NMR (DMSO): 1.08 - 1.20 m, 12 H (CH 3 , iPr); 3.7 - 4.7 m, 9 H (HCO, H 2 CF); 7.19 bs, 2 H .NHa- ϊ 8.09 s, 1 H (H-S, Ade); 8.15 ε, 1 H (H-2, Ade). Bromotrimethylsilane (2.17 ml, 16.4 mmol) is added at room temperature to a solution of this compoLtnd (1.6 g, 4.1 mmol) in acetonitrile (8 ml) and the mixture is stirred for 24 h at room temperature. The reaction mixture is concentrated in vacuo to a minimum volume, the residue is codistilled with toluene (2 x 10 ml) and then with methanol (10 ml) and 2% aqueous ammonia solution (10 ml) is added. After 1 h at room temperature, the solution is evaporated to dryness, the residue ie dissolved in a minimtim amount of water and the solution is applied onto a column of cation-exchanger Dowex 50 (H+-form, 50 ml). The column is washed with water to negative reaction to bromides and the product is eluted from the column with 2% aqueous solution of ammonia. The obtained solution is concentrated in vacuo to a minimum volume and the residue is applied onto a column of anion-exchanger Dowex 1 (acetate form, 50 ml). The coltimn is washed with water and the product is eluted with 1M acetic acid. Evaporation in vacuo and codistillation of the residue with water affords (R)-9-(3-fluoro-2-phoεphonomethoxypropy1 )adenine (1.14 g, 91%), m.p. 194-197 0C, COCDD +18.68 o ( c 0.4, water). For C9H13FNSQ P (305.2) calculated: 35.41% C, 4.29% H, 6.22% F, 22.94% N, 10.14% P; found: 36.65% C, 5.34% H, 5.77% F, 20.14% N. 9.79% P. Example 5 A mixture of 2-amino-6-chloropurine (1.75 g, 10.3 mmol), sodium hydride (60% suspension in paraffin oil, 376 mg, 9.4 mmol) and dimethylformamide (25 ml) is heated at 90 °C for 5 min. (R)-l-Fluoro-2-(diieopropylphoεphonylmethoxy)-3-(p-to¬ luene sulfonyloxy)propane (4.0 g, 9.4 mmol) is then added and the mixture is heated under stirring at 90 &C for 2 h (the reaction is monitored by TLC in chloroform-methanol 5:1). The solvent iε evaporated at 40 c*C and 100 Pa, the residue iε mixed with Celite (25 ml) and chloroform (150 ml) and the obtained εu.spensioπ is filtered through Celite. The filtrate is concentrated to a minimum volume and the residue iε chromatographed on a column of εilica gel (150 g) in a mixture of chloroform and methanol (50:1), affording 2.8 g (68%) of (R)~2-amino-9-(2-diisopropylphoεphonylmethoxy-3- fluoropropyl)-6-chloropuriπe, m.p. 133—134 °C (toluene). For C I H 2 - I C1FN E 0 Ξ P (439.S) calculated: 40.96% C, 5.50% H, 8.06% Cl, 4.32% F, 15.92% N, 7.04% P; found: 41.05% C, 5.53% H, 8.48% Cl, 4.40% F, 15.96% N, 7.20% P. MS, m/e (rel.%): 424 (M, 75), 390 (26), 340 (100), 306 (25), 258 (10), 228 (19), 170 (34), 134 (17), 113 (12). A mixture of this compound (i g, 2.27 mmol) and sodium azide (2.0 g) in dimethylformamide (15 ml) is heated at 110 °C for 5 h. The εolvent iε evaporated in vacuo, the reεidue is suspended in chloroform, silica gel (4 g) is added and the mixture iε evaporated to dryness. This material iε applied onto a column of εilica gel (20 ml), the product is eluted with a mixtLire of chloroform and methanol (10:1) and purified by chromatography on a column of silica gel u.sing a mixture of chloroform and methanol (30:1) as eluent. Yield 700 g (69%) of (R)-2-amino-6-azido-9-(2—diisopropy1phosphoπyImetho- xy-3-fIuoropropy1) urine, m.p. 146-148 °C. For Ci5H *FNs0EP (446.36) calculated: 40.35% C, 5.41% H, 4.25% F, 25.10% , 6.93% P; found: 40.45% C, 5.56% H, 4.01% F, 25.23% N, 6.81% P. MS, m/e (rel.%): 445 (M - H, 1) , 431 (100), 405 (2D, 390 (3), 319 (9), 225 (8), 210 (9), 150 (11). A mixture of this compou.nd (150 mg, 0.336 mmol), bromo¬ trimethylsilane (111 _μl, 0.84 mmol) and acetonitrile (1.8 ml* iε stirred at room temperature for 24 h. The reaction mixture is then codistilled with toluene, a dilute aqueous ammonia soluition is added (5:1, 5 ml), the solution iε concentrated in vactio, the residtie is codistilled with water, dissolved in the same solvent and the solLition is applied onto a column of cation—exchanging resin (Dowex 50) in H+—form. The column i^ washed with water to negative reaction to bromide ions and the compound is then obtained by elution with dilute aqueous ammonia. After evaporation, the product is dissolved in water and applied onto a column of an anion-exchanging resin (e.g. Dowex 1 ) in acetate form. The column is first washed with water and then the compound is eluted with 1M acetic acid. Evaporation of the solvent and codistillation of the residue with water affords 80 g (69%) of (R)-2-amino-6-azido~9-(3- -fluoro-2-phosphonomethoxyprop l )purine. For C9H12F SO4P (327.2) calculated: 33.03% C, 3.69% H, 34.24% N, 9.46% P; found: 33. i4% C, 3.78% H, 34.50% N, 9.30% P. Example 6 A mixture of (R)-2-amino-9-(2-diiεopropylphosphony1- methoxy-3-fluoropropyl )-6-chloropurine (500 mg, 1.14 mmol) according to Example 5 and 75% trifluoroacetic acid (5 ml) is allowed to react at room temperature for 48 h (monitoring by TLC in chlorofor -methanol 5:1). The reaction mixture is concentrated in vacuo on a rotary evaporator, the residue is codistilled with water, neutralized with a mixture of aqueous ammonia and methanol (1:10, 15 ml) and the solvent iε evaporated to dryneεs. Chromatography on a column of silica gel (Silpearl, 30 ml) in a mixture of chloroform and methanol (20:1) affords 440 mg (92%) of (R)-9-(2-diisopropylphoεpho- πylmethoxy-3-fluoropropyl )guanine, m.p. 165-173 OC. For CiEH2EFNs0 6 P (421.4) calculated: 42.75% C, 5.98% H, 4.50% F, 16.62% IM, 7.35% P; found: 42.95% C, 5.92% H, 4.40% F, 17.16% N, 7.34% P. MS, m/e (rel.%): 420 (M - H, 5), 406 (100), 364, (13), 322 (50), 304 (4), 262 (4), 226 (5), 210 (10), 151 (19). A mixture of this compound (290 mg, 0.7 mmol), bromotrimethylsilane (364 mg, 2.75 mmol) and acetonitrile (3 ml) is stirred at room temperature for 24 h. Then the reaction mixture is codistilled with toluene, the residue is mixed with dilute aqueous ammonia (5:1, 5 ml), the solution is concentrated, codistilled with water and the residuie in aqueoLiε εolution iε applied onto a column of a cation-exchanging resin (Dowex 50) in H+-form. The column is washed with water to negative bromide reaction and the compound iε eluted with dilute aqueous ammonia and applied onto a column of an anion-exchanger (Dowex 1) in acetate form. The column is washed with water and the product iε eluted with 1M acetic acid. Evaporation of the solvents and codistillation with water affords (R)-9-(3-fluoro-2-phospho- nσmethoxypropyl)guanine (210 mg, 95%). For CΪHISFIMEOEP (321.2) calculated: 33.65% C, 4.07% H, 5.91% F, 21.80% N, 9.64% P; found: 33.26% C, 5.09% H, 6.00% F, 21.78% N, 8.47% P. MS, m/e (rel.%): 322 (M + H, 50), 279 (10), 257 (8), 228 (10), 131 (28), 149 (100), 110 (100). Example 7 A mixture of (R)-2-amino-6-azido-9-(2-diisopropoxyphosphonyl- methoxy-3-fluoropropyl )purine (according to Example 5, 300 mg) and 10% palladium on carbon (30 mg) in methanol (6 ml) iε stirred at room temperature for 10 h in hydrogen atmosphere. The reaction mixture is filtered and the solvent evaporated. Chromatography on a column of εilica gel (30 ml) in a mixture of chloroform and methanol (40:1) gave 217 mg (77%) of (R)-2,6-diamino-9-(2-diisopropy1phosphonylmethoxy-3-fluoro¬ propyl)purine. For CιsH26FN 6 0-j.P (404.4) calculated: 44.55% C, 6.48% H, 4.70% F, 20.78% N, 7.66% P; found: 44.35% C, 6.67% H, 4.61% F, 20.53% N, 7.52% P. To a εuspension of this compound (150 mg, 0.37 mmol) in acetonitrile (1.5 ml) is added bromotrimethylsilane (263 JJI, 2.0 mmol) and the mixture iε εtirred at room temperature for 24 h. Then the reaction mixture iε codiεtilled with toluene and dilute aqueous ammonia (10:1, 5 ml) iε added. The solution is concentrated, the residue codiεtilled with water and in aqueous solution applied onto a column of a cation-exchanging resin (Eiowex 50) in H+-form. The column iε washed with water to negative bromide reaction, the product iε eluted with dilute aqueous ammonia and applied onto a column of an anion-exchanger (Dowex 1) in acetate form. The column iε washed with water and the product iε then eluted with 1M acetic acid. Evaporation of solvents followed by codistillation with water affords 50 mg of (R)-2,6-diamino-9-(3-fluoro-2-phosphonomethoxypropyl )purine. For C<?Hi4FN 6 θ4P (320.2) calculated: 33.76% C, 4.41% H, 5.93% F, 26.24% IM, 9.67% P; found: 33.46% C, *.83% H, 5.33% F, 26.04% N, 9.37% P. Example 8 A mixture of (R)-2-ειmino-9-(2-diisopropylphosphonylmethoxy- -3-fluoropropyl )-6-chloropurine (according to Example 5, 500 mg, 1.14 mmol) and 10% palladium on carbon (50 mg) in methanol (10 ml) iε εtirred at room temperature in hydrogen atmoεphere for 4 h. The reaction ie monitored by TLC (chloroform-methanol 5:i). The reaction mixture iε neutralized with a mixture of concentrated aqueous ammonia and methanol (1:10, 10 ml), filtered and evaporated. Chromatography on a column of silica gel (50 ml) εucceεεively in 50:1 and 20:1 mixtureε of chloroform and methanol affords 200 mg (45%) of (R)-2-amino-9-(2-diisoprop lphosphonyl ethoxy- 3—fluoropropyl )pLirine. For CIEH2EFNE04P (389.4) calculated: 46.27% C, 6.47% H, 4.88% F, 17.99% N, 7.95% P; found: 44.71% C, 6.44% H, 4.30 %F, 17.44% N, 8.14% P. To a solution of this compound (80 mg, 0.21 mmol) in acetonitrile (1.6 ml) is added bromotrimethylsilane (280 j l, 2.1 mmol ) and the mixture iε stirred at room temperature for 24 h. After evaporation in vacuo, the residue is codiεtilled with toluene, a dilute aqueous solution of ammonia (1:10, 5 ml) iε added, the solution is concentrated and the residue codistilled with water. An aqueous solution of the residue is applied onto a column of a cation-exchanger (Dowex 50) in H+-fσrm. The column is washed with water to negative bromide reaction and then with aqueous ammonia. The residue after evaporation of the ammonia eluate is applied onto a column of an anion-exchanger (Dowex 1) in acetate form, the column iε washed with water and the prodLict is eluted with 1M acetic acid. Evaporation of the solvents and codistillation with water affords 50 mg (80%) of (R)-i-amino-9-(3-fluoro-2-phos- iO - phonomethoxypropyl)purine. For C<?H±3F E04P (305.2) calculated: 35.41% C, 4.29% H, 6.22% F, 22.94% N, 10.14% P; found: 35.59% C, 4.40% H, 6.40% F, 23.15% N, 10.30% P.

Claims: Claims 1. Di(2-propyl) esters of 1-fluoro-2-phσsphonomethσxy-3-p- tolυenesulfonyloxypropanes of the general formula I CHaCβtLjSOzOCHz (I) where the absolute configura ion at carbon atom C2 is B, S or BS. 2. The method of preparing compounds of the formula I according to Claim 1, which is characterized in that 1- luσro-2,3-prσpanediols of the general formula II FCHzCHCHzOH (II) OH where the absolute configura ion at carbon atom C2 is B, S or BS. are reacted with p-toluenesulfony1 chloride in the presence of a tertiary amine, preferably triethylamine or pyridine, in an inert organic aprotic solvent, preferably benzene or dichloromethane, or only in pyridine, the obtained 1-fluoro-3~p-toluenesulfonyloxy-2-propanols of the general formula III FCHzCHCHzOSOzCβliiCHa (III) OH where the absolute configuration at carbon atom C2 is B, S or BS, are reacted with dimethoxymethane in the preβence of phosphorus pentoxide in an inert organic solvent, preferably dichloromethane, the resulting 1-fluoro-2-methoxymethoxy-3- p-toluenesulfonyloxypropaneβ of the general formula IV CHa CβH-i SOz OCHz CHCHz F ( IV) OCH2 OCH3 where the absolute con iguration at carbon atom C2 is B, S or BS, are treated with acetic anhydride in the presence of a Lewis acid, preferably boron trifluoride etherate, at -5 to +5 °C, the obtained 2-acetoxymethoxy-l- luoro-3-p-toluenesulfonyl- oxypropanes of the formula V CHaCβϊLiSOzOCHzCHCHzF (V) OCH2OCOCH3 where the absolute configuration at carbon atom C2 is B, S or BS, are heated with bromotrimethylsilane to 100 - 120 °C, preferably in toluene, the volatile material is evaporated in vacuo and the residue is heated with tri(2-propyl) phosphite at 100 - 120 °C, and the compounds of the general formula I are isolated, preferably by chromatography. 3. The utilization of compounds of the formula I according to Claim 1 in the preparation of N-(3-fluoro-2-phosphonomethoxy- propyl) derivatives of heterocyclic purine and pyrimidine bases of the general formula VI B-CH2CH-0CH2P 0)(0H) 2 (VI) ! CHzF where the absolute con iguration at carbon atom C2 is B, S or BS and B is a substituted purin-9-yl, purin-7-yl, pyrimidin-1-yl or pyrimidine-3-yl moiety or their aza or diaza analogues, which is characterized in that compounds of the formula I are reacted with a sodium salt of the substituted purine or pyrimidine heterocyclic base, its aza or diaza analogue, or their acyl, preferably benzoyl, derivatives, or with a mixture of these bases with an alkali metal carbonate, preferably cesium carbonate, in dimethyl ormamide at 60 - 140 °C, whereupon the solvent is evaporated in vacuo and the residue is treated with Bodium methoxide in methanol and then with bromotrimethylsilane in an inert organic solvent, preferably in acetonitrile, and the resulting compounds of the general formula VI are isolated by chromatography, preferably on ion- xchanging resins.

Inventors: HOLY ANTONIN, JINDRICH JINDRICH, HOLY, ANTONIN, JINDRICH, JINDRICH
Applicants: CESKOSLOVENSKA AKADEMIE VED
Application year: 1992
Main classification: C07F9
IPC codes: C07F9