JPH0198A - Method for producing 2',3'-deoxynucleoside - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION BACKGROUND OF THE INVENTION 11 Separation The present invention relates to an improved process for producing 2',3'-dideoxynucleosides. Furthermore, the present invention provides novel α-(D)-
and α- and β-(L)-2′,3′-dideoxynucleosides.

Typically, 2',3'-dideoxycytidine (dde)
is 1+! synthesized from 2'-deoxycytidine! 0 This is a general method for synthesizing 2',3'-dideoxynucleosides. However, the starting materials for this synthesis are very expensive and not available in large quantities, and the reagents required for this deoxygenation are also very expensive.

Other preparation methods include the use of cytidine or protected cytidine, which is then di-deoxygenated in stages.

The original synthesis of Nor%1i tz 3 used cyclonucleosides as intermediates and involved many steps.

A racemic mixture of β-(D,L)-2',3'-dideoxyadenosine (ddA) was prepared by Robins (M,J).

Robbins)'.

Many of the other procedures involve reduction of the nucleoside to a 2',3'-unsaturated intermediate, which is then catalytically hydrogenated by Perkow et al.
) More complex methods involving reactions have also been described. Many of these methods are summarized in US Patent No. 12. More recently, the photoreductive synthesis of dideoxynucleosides from nucleosides has also emerged.
',3'-dibuosiuracil, but this is easily converted to dde.

Finally, the synthesis of dideoxynucleosides (which are racemic and also anomeric) from various tetrahydrofurans by photochemical chlorination is described in Rocha Reference 15. Cytidine is an expensive starting material for thousands of preparations of ddc. Furthermore, although the substituted tetrahydrofuran used by the Rocha authors is rather inexpensive, it is racemic and an anomeric mixture, yielding dde as a mixture of enantiomers that is difficult to separate. Also,
L-dideoxynucleosides are not available from natural nucleosides. This is the only method known for their production.

SUMMARY OF THE INVENTION In summary, the present invention provides a 2'
, a method for producing 3'-dideoxynucleosides, novel 2',3'-dideoxynucleosides produced thereby, and their effectiveness as antiviral and antibiotic agents, especially against acquired immunodeficiency syndrome (AIDS) infectious agents. This includes its use as an antiviral agent.

An earlier application, US Patent Application No. 028.817, filed March 20, 1987, broadly discloses the method herein presented as the invention. More particularly, the description of the invention and the original schemes I and II shows structural formulas that clearly show that the products produced by the process of the invention can have a base in the β-position or in the α-position. However, these formulas do not so clearly indicate the stereomorphic orientation for the hydroxymethyl moiety pendant to the bentofuranose ring in the 4-position; the actual example in the original application is from (D)-γ-carboxy T-butyrolactone Figure 2 illustrates 2',3'-dideoxynucleosides produced according to the starting invention. We have now illustrated a method for preparing each of the possible enantiomers separately, starting from (L)-γ-carboxy-γ-butyrolactone. The method according to the invention involves chiral synthesis of each enantiomer and avoids the additional step of separating the enantiomers.

In another aspect, the invention includes (L)-dideoxynucleosides. In particular, the present invention includes (L)-dideoxynucleosides prepared by the method of the present invention.

In yet another aspect, the present invention includes a formula useful in the method: (wherein A is an 0-active group as described below, and
, CI, Br and 11, preferably a leaving group selected from CI and Br).

DETAILED DESCRIPTION OF THE INVENTION The present invention is a process for preparing dideoxynucleosides of the formula: wherein B is a base selected from the group of purine and pyrimidine bases, and R is H and a hydroxy protecting group. (a) a member of the group consisting of Noshi- and D-r-carboxy-T-butyrolactone of the formula, wherein R is a hydroxy-protecting group; (b) converting the intermediate from step (a), formula 2, to the group -methyl-T-butyrolactone, of formula 5-0;
-Hydroxy protecting group-methyl-2,3-dioxy-D-
or -L-bentofuranose, the intermediate from step (c) step), formula 3, is converted into a compound of formula 3, where R is a hydroxy protecting group and A is alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl , arylthiocarbonyl, alkylsulfonyl, arylsulfonyl and carbonate groups, the alkyl moiety can be an unsubstituted or substituted C8-C3 alkyl group, and the minor moiety is an unsubstituted or substituted phenyl group. and the substituents on the alkyl and aryl moieties are 〇-activated groups which can be selected from 1 to 3 groups selected from halo and CI to C, alkoxy groups). converting the group -5-0-protecting group -2,3-dideoxy-D- or -L-bentofuranose, (d) converting the intermediate from step (0), formula 4, into a formula HX
and TMSX, wherein R is a hydroxy protecting group and X is FlCl,
1-Leaving group -5-0-protecting group -2,3-dideoxy-〇- or - which is a leaving group selected from Br and I)
(e) converting the intermediate from step (d), formula 5, to an activated base selected from purine and pyrimidine bases pendant on the nucleus of the given base; Reaction with a base activated by reaction of amino and hydroxy groups with an activated compound selected from silylating and acetylating and benzoylating agents in the presence of one of a Bronsted acid and a Lewis acid. and (f) recovering the 2',3'-dideoxynucleoside from the step (e).

A method for producing two representative dideoxynucleosides according to the present invention is schematically illustrated in Scheme 3.

Scheme I The starting material, γ-carboxy γ-butyrolactone, can be conveniently obtained from L- or D-glutamic acid by use of conventional techniques reported in the chemical literature '6.

A suitably protected 2,3-dideoxybentofuranose, 3, is produced by a combination of chemical reactions. This compound and its derivatives are known in the literature.

Thus, in the conversion of the starting material γ-carboxy γ-butyrolactone to 5-0-hydroxy protecting group-methyl-γ-butyrolactone, step (a) involves the reaction of the γ-carboxy group to a hydroxymethyl group, then Reactions with hydroxy protecting group reagents are included.

For example, reduction of the γ-carboxy group and protection of the resulting γ-hydroxymethyl group with a benzyl group (PhCH,-), step (a), can be applied to BH3·SMe of the starting compound of formula l.
, and 16+1′? achieved by sequential reaction with Ph CHt Br? .

Primary alcohol functional groups can be ethers such as trialkyl or dialkylaryl or diarylalkyl or triarylsilyl unsubstituted and substituted benzyl, unsubstituted and substituted alkyl, or as allyl ethers, or esters such as benzoyl, mesitoyl,
It can be protected as pivaloyl, unsubstituted or substituted acetyl, or carbonate. For a more detailed explanation of protecting groups and their associated chemistry, see Protective Groups in Organic Synthesis.
s in Organic 5 synthesis)
J + Greene (T, W, Greene) + John
John Wiley, New Yo
rk).

See 1981. In a more preferred embodiment, a benzyl group, most preferably a benzoyl group, was used as the hydroxy protecting group for the primary alcohol group in the 5-position due to their stability and well-known preparation.

5-0-hydroxy protecting group -methyl-
Conversion of T-butyrolactone to 5-〇-hydroxy-protecting group-2,3-dideoxybentofuranose with formula 3 is accomplished by combining an intermediate with formula 2 with NaH and HCOzBt
Is was then achieved by reaction with H(11).

The present invention uses any one of a number of reducing agents to
It will be understood by those skilled in the art that either or both of the steps (i.e., sequentially) or simultaneously can be performed using other useful methods other than BH2.SMe for use in steps (a) and (b) above. NaB is the reducing agent
H4, NaB Ha and LiCj! or A(l C1
3 or B F 2 , LI A I Ha, Li A
n! H(OMe)s, Li AIH(0-t-Bu
)s, (Sia)z Bl ((dialkyranes) and other dialkylboranes, etc.) It is preferred to use (Sia)t B H as the reducing agent for convenient handling and reactivity.

The conversion of step (c) of the intermediate of formula 3 to an intermediate of formula 4 with an "activated" hydroxy group in the C(1) position of the 2,3-dideoxybentofuranose ring system is:
This hydroxy group is converted into a group that can be easily substituted by Cl or Or by reaction with HCβ or HB r s or preferably TMSBr or TMSCl(+) with a catalytic amount of TMSI (rTMsJ represents a tetramethylsilyl group). This can be accomplished with any reagent effective to effect the conversion. Groups that are easily substituted include alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, arylthiocarbonyl, alkylsulfonyl, arylsulfonyl oyohicarbonate groups (however, the alkyl moiety is unsubstituted or substituted C+-C
s alkyl group, the aryl moiety can be an unsubstituted or substituted phenyl group, and the substituents on the alkyl and aryl moieties are from 1 to 3 groups selected from halo and C1-C3 alkoxy groups. (can be selected) is included. More preferably, the activating group is (as defined above) the corresponding acetoxy and benzoyloxy group,
Most preferably selected from acetoxy.

Step (c) was conveniently carried out using an acetic anhydride/pyridine reagent.

Replacement of the 1-0-activating group functional group in step (6) with a leaving group selected from C1b and Dr converts intermediate 4 into HCI! (
or) IBr) in CH2ci, at medium to low temperatures, and the furanosyl halide 5a or 5 is present in solution as a mixture of anomers (α and β).
b was obtained.

In step (e) of the method according to the invention, said step (d)
) where the base is selected from purine and pyrimidine bases, such bases being well known for obtaining silylated, acetylated or benzoylated or other acylated bases. A suitably activated base activated by reaction with an activating reagent and optionally a Lewis acid such as boron halides, aluminum halides, titanium halides, tin chloride in the presence of a suitable polar or non-polar solvent. (■), in the presence of zinc halides, trimethylsilylbromide, ioside, triflate and other species well known to be used in glycosylation reactions, or in the presence of Brønsted acids such as hydrogen chloride, hydrogen bromide or The reaction was carried out in the presence of hydrogen iodide. It is particularly useful in this step (e) to use silylated purines and pyrimidines in the presence of non-polar solvents such as benzene, toluene, chloroform, dichloromethane, dichloroethane and carbon tetrachloride. Examples of polar solvents used include, among others, eg tetrahydrofuran and dioxane, nitriles such as acetonitrile, dimethylformamide and dimethylsulfoxide. An example of a procedure useful for carrying out step (13) is provided by Brund idge et al.
U.S. Pat. No. 4,625.020 discloses a combination of silylated pyrimidine and 2-deoxy-2-fluoroarabinofuranosyl halide in which the active hydrogens of the hydroxy and amino groups are blocked by a silyl group, such as a trimethylsilyl group. A coupling is disclosed. This reference procedure was applied to step (e) according to the invention, so that a purine or pyrimidine base in which the active hydroxy and amino hydrogens were all blocked by trimethylsilyl groups was reacted with an intermediate having formula 5.

Thus, treatment of the intermediate with formula 5 in CHz CIlz and CHCl5 with the silylated base results in a protected 2',3'-dideoxynucleoside containing a mixture of anomers (β:α=1=1-3: 1) was obtained in good yield (about 4.9% to about 74%).

Alternatively, in other embodiments according to the invention step (c)
When the group "rA" in is acetyl, the intermediate from step (c) is reacted with an activated base in the presence of a Lewis acid as in step (e) to form dideoxynucleosides without passing through step (c). can get things.

After the coupling reaction of step (e), step (f) is performed to obtain a final product that generally has antiviral, antimetabolite, antitumor, anti-human immunodeficiency virus (anti-HIV) activity.
The process according to the invention includes 5-
〇-An additional step is included to remove the protecting group. Suitable procedures for removing this protecting group are well known in the art relevant to the present invention, examples include those described by Greene (T, W.
``Protecting Groups in Organic Synthesis'' by John Y! j-(John Wiley, New York
), 1981.

When the protecting group is a particularly preferred benzyl or benzoyl group, it is subjected to catalytic hydrogenation (H2/Pd C
1z ) or ammonia in methanol (N Hz
/MeOH).

The 2',3'-dideoxynucleoside product (unprotected or protected) is collected from the step (el) or the reaction mixture is collected from an additional step to remove the 5-0-protecting group, and the insoluble reactants and additives are removed from Celite. The resulting product was conveniently recovered by purification by column chromatography, for example on a silica gel column using a mixture of about 1-5% methanol in chloroform as the eluent.

The resulting anomers (β and α) were separated using chromatographic and crystallization techniques well known in the art relevant to the present invention. The desired active β-anomer was generally obtained in larger proportions than the apparently biologically inactive, or at least less biologically active α-anomer.

Therefore, the method according to the invention is (D)-β-1(D)-α
-2' containing (L)-β- and (L)-α-compounds
, provides a stereospecific synthesis for producing 3'-dideoxynucleosides.

The following is a general procedure for separating β and α anomers in which a mixture of Niazmers is applied to a silica gel HPLC column and a gradient mixture of ethyl acetate and hexane is applied at high pressure (100
Elute at a flow rate of ~500 ml/min). The pure anomer is obtained by pooling the appropriate portions and removing the solvent, for example by evaporation under reduced pressure.

2', 3' produced by the method of the present invention as described above
- The base component B of the dideoxynucleoside is selected from purine and pyrimidine bases.

Representative components when derived from purine bases are: 6-aminopurin-9-yl (adenin-9-yl), 2-aminopurin-9-yl, 2,6-diaminopurin-9 -yl, 2-amino-6-hydroxypurin-9-yl (guanine-9-yl), 6-hydroxypurin-9-yl, In addition to the above, component B is 2-halopurin-9-yl, 6-yl
Halopurin-9-yl or 2,6-cyhalopurine-9
-yl, in which case the base component is a step (
There is no need for activation, eg complete silylation, to carry out the condensation or camping reaction in e). Representative components when derived from pyrimidine bases are: 2.4-dihydroxypyrimidin-1-yl, 5
-Methyl-2,4-dihydroxypyrimidin-1-yl, 5-ethyl-2,4-dihydroxypyrimidin-1-yl, 2-hydroxy-4-aminopyrimidin-1-yl, 5-vinyl-2,4- Dihydroxypyrimidin-1-yl, 5-halovinyl-2,4-dihydroxypyrimidin-1
-yl, 5-halomethyl-2,4-dihydroxypyrimidine-1
-yl, 5-haloethyl-2,4-dihydroxypyrimidine-1
-yl, the 5-methyl and 5-ethyl substituents are representative 5-yl,
It is an alkyl substituent, and the 5-vinyl substituent is a typical 5-vinyl substituent.
-alkenyl substituent. 5-Halobinyl (Ma or 5
-Haloalkenyl) Base (7) Examples of halo groups include 1 to 3
or 4 F, CI and Sr groups.

The method according to the invention is therefore useful for the production of various 2',3'-dideoxynucleosides, both pyrimidine and purine nucleosides, which have antiviral, antimetabolite and antitumor activity, as well as activity against the human immunodeficiency virus.

The following examples are illustrative of but some representative embodiments of the method according to the invention and are not to be construed as limiting the scope.

All parts and percentages are by weight and temperatures are degrees Celsius unless otherwise indicated.

The following scheme (■) relates to Examples 1-9.

Ku ＝　　　　　　　　　- /-one \.

Examples Example 1 l-O-acetyl-5-0-pene, dil-2,3 depletion f
5-0-benzyl-2,3 prepared according to Noe (II) literature IS+lI
-dideoxy-D-bentofuranose (1) 1.00
The reaction mixture was stirred for 2.5 hours at room temperature, stirred on ice, stirred and extracted with dichloromethane. The organic layer was washed with IN hydrochloric acid, saturated aqueous sodium bicarbonate and brine. The dichloromethane solution was dried over sodium sulfate, filtered and evaporated to dryness to yield 1-0-acetyl-5-0-benzyl-2,
3-dideoxy-D-bentofuranose 1.20g (1
00%) was obtained as a colorless liquid. Chromatography on silica gel with a mixture of ethyl acetate and hexane gave pure compound (II). Nuclear magnetic resonance spectra supported the desired structure and showed that compound (n) was a 3:2 mixture of anomers.

However, the crude compound (II) was essentially pure and was usually used directly in the next step.

Example 2 5-0-benzyl-2,3-dideoxy-D-/pentofuranosyl chloride (Illa) 1-0-acetyl-5-0-benzyl-2,3-dideoxy-〇-bentofuranose 0.128 A 0.125N solution of hydrogen chloride in dichloromethane is added to a solution of g (0,00051 mol) in dichloromethane 1 tel at 0° C. 4 ya
j! (0,00051 mol) was added.

After 1 hour at 0°C, nuclear magnetic resonance spectra of a portion of this solution showed complete conversion to moisture sensitive 5-0-benzyl-2,3-dideoxy-D-pentofuranosyl chloride (Ina). The above solution was used for direct coupling tests.

Example 3 5-0-benzyl-2,3-dideoxy-D-pentofuranosylbromide ([[b) 1-〇-acetyl-5-0-benzyl-2,3-dideoxy-〇-bentofuranose 0.0965 g(0,0
00384 mol) in 1 ml of dichloromethane
0.3 of hydrogen bromide in dichloromethane to solution at °C
A 6N solution of 1 ts Il (0,00036 mol) was added. After 15 minutes, nuclear magnetic resonance spectra of a portion of this solution showed complete conversion to moisture sensitive 5-0-benzyl-2,3-dideoxy-D-pentofuranosyl bromide (mb). The above solution was used for direct coupling tests.

Example 4 5'-〇-benzyl-2',3'-dideoxyuridine (IV) Uracil 0.24 g (0,00214 mol), hexamethyldisilazane 0.52 tel (0,0024
7 mol) and ammonium sulfate 0.025 g (0
,00019 mol) in dry acetonitrile 8111 was refluxed under argon for 2 hours. The mixture was cooled, evaporated to dryness, and the solvent was changed to dry dichloromethane to give 5 all o,o-bis-trimethylsilyluracils.
．． A 428M solution resulted. 5-0-benzyl-2,3-
Dideoxy-D-pentofuranosylbromide 0.104
g (0,000384 mol) of dichloromethane 2r
A 0.428 M solution of bis-trimethylsilyluracil in dichloromethane to a solution at 40 °C in al.
m 11 (0,000856 mol) was added and the temperature was gradually raised to 25°C. After an overnight period at room temperature, react with 0.2 mA of methanol! quenched with water, filtered through Celite, and washed with 30:1 dichloromethane-methanol.

The crude product was chromatographed on silica gel with 75% ethyl acetate in hexane to separate 5'-0-benzy-2',3'-dideoxyuridine and its C(1) epimer (α-anomer). Mixture (β:α=2:
1) 0.105g (90%) was obtained.

Nuclear magnetic resonance spectra supported the structure.

The β and α anomers are separated by chromatography on silica gel 20°2'.

Example 5 2',3'-dideoxyuridine (V) 0.425 g (0,00024 mol) of pre-reduced palladium chloride and 5'-〇-benzy-2',3'-dideoxyuridine (α and β anomers) 0.080 g (0.00
A suspension of 0.0265 mol) in 1.5++4 dry methanol was hydrogenated for 1 hour with 15 p,s,i, of hydrogen in a Parr shaker. The catalyst was removed by filtration through Celite and the filtrate was washed with water.
ex) Neutralized with 0.70 g of W GR resin for 15 minutes at room temperature, filtered again and evaporated. The crude product was purified by silica gel chromatography eluting with 5% methanol in dichloromethane to yield 2',3'-dideoxyuridine as well as the α anomer (β:α = 2:1).
0.0545g (97%) was obtained.

The nuclear magnetic resonance spectra of the main components of the mixture agreed with the literature data +1%.

Example 6 5'-〇-benzyl-2',3'-dideoxytidine (VI) Cytosine 0.363 g (0,00327 mol), Hexamethyldisilazane 0.78 ml (0,0037
mole) group and ammonium sulfate 0.038 g (0,
A suspension of 0.00029 mol) in 10 ml of dry acetonitrile was refluxed for 2 hours under argon. Cool the mixture;
Evaporating the solvent and precipitating the solution into 5 ml of dichloromethane yields O,N-bis-trimethylsilylcytosine.
．． A 654M solution was obtained.

A 0.654 M solution of bis-trimethylsilylcytosine in a solution of 0.92 g (0.0004 mol) of 5-0-benzyl-2,3-dideoxy-D-pentofuranosyl chloride in 4 ml of dichloromethane at 0° C. 1.2
6 ml (0,00082 mol) were added and the temperature was gradually raised to 25°C. After an overnight period at room temperature, the reaction was quenched with 0.21111 l of methanol, filtered through Celite and washed with 20:1 dichloromethane-methanol. The crude product was chromatographed on silica gel eluting with 5% methanol in methylene chloride to give a 5'-0
- a mixture of benzyl-2',3'-dideoxycytidine and its C(1) epimer (α-anomer) (β:α
=11) 0.090g (74%) was obtained. Nuclear magnetic resonance spectra supported the assigned structure. The β and α anomers were resolved on silica gel, but the separation of the anomers was not yet complete.

Example 7 2',3'-dideoxycytidine ■) Pre-reduced palladium chloride 0.070 g (0,000395 mol)
and 5'-〇-benzy-2',3'-dideoxycytidine (α and β anomers) 0.040 g (0
,000132 mol) in 1.5 d of dry methanol was hydrogenated for 1 hour with 15 p, s, i, of hydrogen in a Parr shaker.゛The catalyst was removed by filtration through Celite, and the filtrate was filtered through A+++berlite.
Neutralized with 0.1 g of IRA resin for 15 minutes at room temperature, filtered again and evaporated. The crude product was dissolved in dichloromethane with 2
Purification by silica gel chromatography eluting with 5% methanol yielded 0.020 g of 2',3'-dideoxycytidine and the α anomer (β:α = 2:1).
(71%) was obtained.

Nuclear magnetic resonance spectra of the main components of the mixture are based on literature data 5
matched.

Example 8 5'-〇-benzyl-2',3'-deoxythymidine (■) Thymine 0.252 g (0,002 mol), hexamethyldisilazane 0.5 ml (0,0024 mol) and -
A suspension of 0.01 g (0,0001 mol) of dotrimethylsilane was refluxed for 48 hours and then cooled. This o
, a solution of 0-bis-triethylsilylthymine is 5-0-
A solution of 0.271 g (0.001 mol) of benzyl-2,3-dideoxy-D-pentofuranosyl bromide in 12.1 mji of dry chloroform at 0)-78 DEG C. was added.

The mixture was gradually allowed to reach room temperature and stirred overnight. The reaction was quenched with 0.5 m methanol, diluted with dichloromethane, washed with water and brine, and dried over sodium sulfate.

The crude product was chromatographed on silica gel eluting with 4=1 ethyl acetate/hexane.

The yield of 5'-〇-benzy-3'-deoxythymidine (both anomers were obtained, α:β = 1:1) was 0.180
g (58%).

Nuclear magnetic resonance spectra supported the structure. Although no separation of the anomers was performed, chromatographic methods are well used for their separation in nucleoside chemistry 26+Z+.

Example 9 5'-0-Benzyru 2', 3'- Dideoxyadenosine (IX) Adenine 0.270g (0,002 mol), hexa.

0.5 mff1 (0,0024 mol) of methyldisilazane and 0.01 g of iodo5-trimethylsilane (
0,0001 mol) was refluxed for 18 hours and then cooled.

A solution of this N,N-bis-trimethylsilyladenine was mixed with 0.271 g (0,001 mol) of 5-0-benzyl-2,3-dideoxy-D-pentofuranosylbromide in 12.1 d of dry chloroform. Added to solution at 78°C. The mixture was gradually allowed to reach room temperature and stirred overnight. The reaction was quenched with 0.5 wl of methanol, diluted with dichloromethane, washed with water and brine, and dried over sodium sulfate. The crude product was chromatographed on silica gel eluting with 8% methanol in dichloromethane. 5'-〇-benzyru 2', 3
The yield of '-dideoxyadenosine (β:α=3:1) was 0.158 g (49%).

Nuclear magnetic resonance spectra supported the structure. Although the separation of the anomers shown in Scheme II is not yet complete, it can be achieved by chromatographic methods typically used in nucleoside chemistry.

The following scheme (■) relates to Examples 10-14 and 16 and 17. The biological activities of representative new compounds are shown in the table, Biological Activity Against Human Immunodeficiency Virus (HIV).

Example 10 (L)-5-0-benzoyl-2,3-dideoxypentofuranose (X) 0.5 M solution of dicyamylborane in THF solution 20
(L)-5'-benzoyloxy-5-hydroxymethylbutyrolactone under nitrogen at 0°C to 0d 15.0
g (0,068 mol) of dry tetrahydrofuran 4
A solution in Od was added dropwise. After 20 minutes at 0°C the reaction mixture was warmed to 22°C. The reaction mixture was stirred at 22° C. for 1 z h and then worked up by slowly adding 12 Jnl of water and refluxing for 30 min. The reaction mixture was cooled to 0<0>C and 30% hydrogen peroxide 24- was slowly added maintaining pi at 7-8 with IN sodium hydroxide. After the addition, the reaction mixture was evaporated on rotovapor at 30° C. under reduced pressure to give an oily residue. This was partitioned between 500° of dichloromethane and 150° of water. The aqueous layer was extracted with 2×100 − of dichloromethane, then the combined organic layers were washed with 50 − of water, dried over anhydrous sodium sulfate, and evaporated to give an oil. (L)-5-
Yield of 0-benzoyl-2,3-dideoxypentofuranose = 15.0 g (99%). 'H-NMR supported the structure and the oil was used directly in the next step.

Example 11 1-0-Acetyl-(L)-5-〇-Penzoyl-nye 2-poor lj'Nojin iotΣ2jノ: Gosagi■Ω--(L)-
5-0-benzoyl-2,3-dideoxypentofuranose (12.0 g, 0.054 mol) in pyridine 24
- and acetic anhydride 12- were stirred at 22° C. for 2 hours, then 100 g of ice.

It was then treated with 600+d of ethyl acetate. The reaction mixture was then washed with 3X100a+1 of IN hydrochloric acid, 4X100d of saturated aqueous sodium bicarbonate and 100sd of brine. The organic layer was dried over sodium sulfate and evaporated to give a pale yellow oil. This can be used as is or on silica gel 35%...→50% EtO
Can be chromatographed with Ac/hexane. Yield 1: 8.6 g of 1-0-acetyl-(L)-5-0-benzoyl-2,3-dideoxy-bentofuranose (
60%).

The NMR supported the structure. Other fractions contained the desired material, but were impure.

Example 12 α,β-(L)-5′-0-benzoyl-2′,3′
-dideoxyadenosine (XI) 1-0-acetyl-(
L)-5-0-benzoyl-2,3-bentofuranose (2,97 g.

0,01125 mol) in 20r
Dissolved in rf. To this was added 1.
44ml! added. After stirring this for 20 minutes at 22°C, 28.93 ml of a 0.428M solution of bissilyladenine in 1,2-dichloroethane was added.

The solution was stirred at 22°C for 120 hours. The reaction mixture was then cooled to 0° C. and then treated by dipping into 50° C. of cold saturated aqueous sodium bicarbonate. The reaction mixture was partitioned between 300mf dichloromethane and 2x50rn1 water. The organic layer was dried and evaporated to give a product mixture, 3.3 g (87%) of a colorless oil.

Example 13 α and β anomers of 9-(L)-5′-0-benzoyl-2′,3′-dideoxyadenosine (XI[[,
A and B) This material is equilibrated and eluted with 60% Me OH/Hz O as solvent Waters C
-18 Prep Baku Cartridge with Waters (W
aters) LC-500 chromatography system. Dissolve the sample in 4 ml of methanol,
Injected. Elution was performed using a refractive index detector at 0.11/min.

Both fractions were collected and analyzed by HPLC. Pool similar, evaporate to 1/3 volume, dichloromethane 3X
500-. The pooled a-layer was dried and evaporated to yield 660 m of pure 9-(β)-(L)-5'-0-benzoyl-2',3'-dideoxyadenosine.
g (NMR supports structure) and pure 9-(α)-
(L)-5'-〇-benzoyl-2',3'-dideoxyadenosine 560 mg [NMR supports structure]
was gotten.

Also, 9-(α,β)-(L)-5'-0-benzoyl-
A mixture of 2',3'-dideoxyadenosine (core 1.8
g) was obtained.

Example 14 9-(β)-(L)-2',3'-dideoxyadenosine (XIV A) 9-(β)-(L)-5'-0-benzoyl-2',3
'-dideoxyadenosine (270 mg.

0.0008 mol) was dissolved in methanol 5d. To this was added 20 d of methanol saturated with ammonia. This was plugged and vented into a bubbler. The reaction was completed after 16 hours and then evaporated. The residue was dissolved in dichloromethane.
Chromatographed on a silica gel column packed and eluted with 0% methanol. The gradient is 2 in dichloromethane.
Proceed to 0% methanol. pool the desired net percentage;
Evaporated. When the residue is crystallized from methanol, 9
160 mg (85%) of -(β")-(L)-2',3'-dideoxyadenosine was obtained. (NMR supported the structure.) Calculated values: C51,05, H5,57, N29 .7
7 Measured values: C51.15, H5.63, N 29.
679-(α)-(L)-2', 3'-dideoxyadenosine (XIVB) 9-(α)-(L)-5'-0-benzoyl-2',
3'-dideoxyadenosine) (250■, 0.000
74 mol) was dissolved in methanol 5-. To this was added 20-methanol saturated with ammonia. This was capped and vented into a bubbler.

After 16 hours, the reaction mixture was evaporated and chromatographed on a silica gel column packed and eluted with 10% methanol in dichloromethane. The gradient progressed to 20% methanol in dichloromethane. The desired pure fractions are pooled, evaporated, and then crystallized from methanol to yield 9
-(α)-(L)-2',3'-dideoxyadenosine 170 g (98%) was obtained ('HNMR supported the structure.) Calculated values: C51,05, H5,57, N 29.7
7 Measured values: C51.04,)I 5.59, N2
9.64 Example 15 α,β-(D)-5'-0-benzoyl-2',3'-
The mixture containing 9-(α,β)-(D)-5′-0-benzoyl-2′,3′-dideoxyadenosine from the coupling reaction of dideoxyarnosine (XVA) was purified by Waters blep.・Baku C-18
50% MeOH/H! for column and elution. Reversed phase chromatography was performed on a Waters LC500 chromatography system using O. This material was dissolved in 2 ml of methanol and injected. 0.1ε/
A flow rate of minutes was used. After eluting solvent 1.51, mobile phase 41 was slowly eluted with 60% MeOH/HzO.

Both fractions containing primarily the α-(D)-5'-0-benzoyl isomer were then evaporated. The residue was then rechromatographed on a 1" x 14" flash column packed with C-18 and eluted with a gradient of 50% methanol/water to 65% methanol/water.

Both fractions were collected and the pure fractions were pooled and evaporated to 1/3 volume. This was then extracted with 5×100 dichloromethane and the organic phases were pooled, dried over anhydrous sodium sulfate and evaporated. When this substance is crystallized from acetone, α
-(D)-5'-0-benzoyl-2',3'-dideoxyadenosine was obtained. (NMR supported the structure.) (D) -XV B NH.

9-α-(D)-2′,3′-dideoxyadenosine (XV B) Substance 9-(α)-(D)-5′-0-benzoyl-2
',3'-dideoxyadenosine 220■ (0,000
65 mol) of methanol 8WIl saturated with ammonia
dissolved in. This was stirred in a closed system for 16 hours.

The reaction mixture was then evaporated and 12
Chromatographed on a silica gel column packed with % methanol and flNm. Both fractions were collected and the pure fraction was pooled and evaporated. Crystallization of the residue from methanol yielded 114 ow (75%) of 9-(α)-,(D)-2',3'-dideoxyadenosine.

('HNMR supported the structure.) Example 16 (α,β)-(L)-5'-0-benzoyl-2',3
'-Dioxycytidine (XLI) 1-0-acetyl-5-(L)-0-benzoyl-2,3-bentofuranose 2.815 g (10.66 mmol) was added to 1.2
-Dissolved in dichloroethane 20-. To this was added 1.8 Mi (1
5,46 mmol) (1,45 eq.) and this
Stirred for 20 minutes at 2°C, then cooled the solid mass to -78°C. This was then added with 50 d (15,65 mmol) of a solution of bissilylcytosine in a volume of 0.31 moJ).
added. The bath was removed and the reaction mixture was warmed to 22°C.

After 16 hours the reaction mixture was worked up by pouring into 50 ml of cold saturated aqueous sodium bicarbonate. This was partitioned with 300° of dichloromethane (cHz Clx) and the organic layer was washed with 50° of water. The organic layer was dried over anhydrous sodium sulfate and evaporated to give α,β-(L)-5'
3.36 g of a mixture containing -0-benzoyl-2',3'-dideoxycytidine was obtained.

Example 17 α,β-(L)-5'-0-benzoyl-2',3'-
The material separated from the coupling reaction of dideoxycytidine (XWA) was dissolved in 35% MeOH/65% pH 6.5.
Reversed phase C-18 chromatography cartridge (Waters C-18 Prep Vac) equilibrated and eluted with 0.01M ammonium phosphate buffer.
Waters L C-50 using
Separated on a 0 chromatography system. column to 50%
MeOH150% pns, s buffer total 6j! 0. It was eluted at lllZ minutes.

Collect 5% MeOH/CH,C using 2 development.
I, analyzed by TLC on 5iO1 plates. T
Both fractions containing mainly the first spot on the LC were pooled and evaporated. For further purification, the material was then 40% M
C-1 eluted with eOH/60% buffer, pn6.5
Flash chromatographed on an 8 column 1" x 14". After pooling the desired pure fractions and evaporating to 1/3 volume, the material was dissolved in dichloromethane (cHz C1z) 3X.
100-. Combine the organic layers and dry
When evaporated, β-(L)-5'-0-benzoyl-2
380 ml of ',3'-dideoxycytidine was obtained. (
NMR supports structure. ] Similarly, both fractions from the LC-500 chromatograph, containing primarily the second spot, were pooled and evaporated. This substance is mixed with 40% methanol/60%
Flash C-1 to fill and elute with pH 6.5 buffer
Purified on 8 columns 1" x 14#. After pooling the desired pure fractions it was evaporated to 1/3 volume and the material was partitioned between 3 x 500 m of dichloromethane. The combined organic layers were dried and evaporated. Upon reaction, α-(L)-5'-〇-benzoyl-2',3'-dideoxycytidine 506■ was obtained. ('HNMR supports the structure.) β-(L)-2',3' -dideoxycytidine (XVIr B) β-(L)-5'-0-benzoyl-2',3'-
Dideoxycytidine (250 mg, 0.00079 mol) was dissolved in methanol 5- and ammonia-saturated methanol 20- was added. The reaction was vented through a bubbler. The reaction mixture was stirred at 22°C for 16 hours. The reaction was completed and then evaporated.

Crystallization of the residue from ethanol/ether gives β-
(L) -2',3'-dideoxycytidine (1
32 mg, 78%) was obtained.

('HNMR supported the structure.) α-(L)-2',3'-dideoxycytidine (XWC-) α-(L)-5'-0-benzoyl-2',3'-dideoxycytidine ( 250mg, 0.00079mol)
was dissolved in 5-methanol and to this was added 20-methanol saturated with ammonia. The reaction was vented into a bubbler. The reaction was completed after stirring at 22°C for 16 hours.

The reaction mixture was then evaporated and triturated with ether leaving an oil. This was crystallized from methanol/ether to yield α-(L)-2',3'-dideoxycytidine (140 mg).

84%) was obtained. [IH NMR supported the structure.

] Example 18 α. β-(D)-5'-0-benzoyl-2',3'-
α,β-(D)-5′-0-benzoyl-2′,3′ of dideoxycytidine (XVIA)
- a mixture of dideoxycytidine in 40% MeOH/p
Flash chromatography was performed on a reverse phase C-18 column (1"X14") packed and eluted with H6,5,0,01M ammonium phosphate buffer. Both fractions were collected and analyzed by TLC on silica gel plates using 2 development and 5% methanol in dichloromethane as solvent. Both parts, mainly including the second spot, are pooled and evaporated,
It was then chromatographed again on the same column in a similar manner. This produced a pure fraction, which was pooled and evaporated. The residue was dissolved in 5% methanol in dichloromethane and filtered through glass wool (to eliminate buffer salts). When the filtrate is evaporated, α-(D)-5'-0
-benzoyl-2',3'-dideoxycytidine 160
mg was obtained. ('HNMR supports the structure.] Also! (During this separation in method 1, pure β-D-dd
c and a mixture similar to that originally started was obtained.

α-(D)-2',3'-deoxycytidine (
XVI B) 80 mg (0,00025 mol) of α-(D)-5'-0-benzoyl-2',3'-dideoxycytidine were dissolved in methanol 5- and then methanol saturated with ammonia 10- was added. The reaction was vented through a bubbler. The reaction was completed after 16 hours and then evaporated.

Trituration of the residue with ether gives α2(D)-2',
50 μ of 3′-dideoxycytidine were obtained. ('
HNMR supports the structure. ] Example 19 9-α-(L)-2′,3′- 9-α-(L)-2′,3′-dideoxyadenosine 45 mg, (0,00019 mol) H distilled to
, 03 ml and adenosine deaminase [Sigma, type n, E (3,5,4,4)
45 mg of J was added. The reaction mixture was stirred at 22°C for 18 hours. At this time, HPLC analysis (20% MeO
H/pl (6,5 ammonium phosphate buffer, C-18
column) showed that the reaction was 97% complete.

The H2O was then evaporated and the reaction mixture was added to the previous step.
Combined with 0 mg scale reaction mixture. Then add it to 10% M
Chromatographed on a silica gel column packed and eluted with eOH/CHz C1.

Fractions were collected, analyzed, and the pure percentages were pooled to yield 9-α-
L-2',3'-dideoxyinosine 42mg (0,
00018 mol) (76% yield) was obtained, melting point>2
50°C, (NMR supported the structure.) Viability to human immunodeficiency virus (HIV), μm/- Reference 1, Samukov et al. (Samukov, V, V, H
Ofitserov+V, 1. , Bioorg, Khim, 1
983. 9. 132゜2, Prisbe et al.
be, E, J, ; Martin.

J,C,), Synthetic Communications (S
ynth, Con++sun, ), 1 9
B5. 1 5. 4 0 1.

3. Horwitz et al.
P, ; Chau, Noel, M, ;D
onatti+J, T, ) * Journal of Organic Chemistry (J, Org, Ches, ), 1967. 32
．． 817゜4, Robbins (M,
J.) and others, Journal of American
Chemical Society (J, As, Che++
+, Soc, ), 1976. 98゜8213゜5, Marumoto et al.
onjo, M, ) + chemical and pharmaceutical buretine (chew, Pharm,
Bull, ), 1974.

22.128°60. Furukawa (Y, ) and others,
Chemical and Pharmaceutical Bulletin (chew, Pharm, Bull,)+
1 9 7 0. 1 8゜554゜7, McCarthy, J, R
,)others.

Journal of the American Chemical Society (J, A-, Ches, 5oc-) + 1966
+88.1549°8, Robins (M, J, ) and others, Tetrahedron Letters (Tet, Lett,
), 19 B 4゜25.367゜9, David et al.
yey, G, L Carbohydrate Research (ca
rbohydrateRes, ), 1980, 82.4
5゜10, Adachi, 'r, et al., Journal of Organic Chemistry (J, Or
gChes, ), 1979. Earthwork, 14α4゜11
, Team Haka (Thiem, J, 1asch,
D, ) *Nucleosides and Nucleos
1985°4.487°12, Verheyden et al.
, P, H,; Moffatt, J, G, ), U.S. Patent No. 3.817.982 (1974).

13, Saito (1.) et al., Journal of the American Chemical Society (J, Am, Chem, Sac,), 1986.
108°3115°14, Lin, T, S, et al., Journal of Medicinal Chemistry (J, Med.

Chem, ), contributed.

15, Kaulina (L, T,
), as well as Kimya Geterosiklisieskik Sedinen (nihim, Geterosikl, 5oedin,
), l 932°101°16, Taniguchi et al.
Koga, K.; Yamada, 3. ),
Tetrahedron, 197
4.30.3547゜17, Ravid et al.
verstein.

RoM,; Sm1th, L, Ro),
Tetrahedron, 19
78. 34. 1449°18, Hubbard
ard, A., J.) et al., Nucleisoc.
Acid Research (Nucleic Ac1dRes)
), 1984, 7.6827°19, Stone et al. (Stone, K-; Little, Ro
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1919゜21, Woenckh House
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゜Procedural amendment (method) 1. Indication of case 1985 Patent 1iII No. 679
No. 78 No. 2, Title of the invention 2', 3'-deoxynucleoside manufacturing method 3, Relationship with the case of the person making the amendment Applicant name: Bristol-Myers Company 4, Agent 5, Date of amendment order: 1988 June 28th (same content)
Koyu ε5! Pickled J

Claims (24)

[Claims] (1) Formula, ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, B is a base selected from the group of purine and pyrimidine bases, and R is a member selected from H and hydroxy protecting groups.) (a) Formula, ▲ Numerical formula, chemical formula, table, etc. ▼ Formula 1 of the group consisting of L- and D-γ-carboxy-γ-butyrolactone. By reacting a compound of formula 1 with a carboxy group reducing agent and then reacting the resulting hydroxymethyl group with a hydroxy protecting group reagent, the formula, ▲mathematical formula, chemical formula, table, etc.▼formula 2 (in the formula, (b) converting the intermediate from step (a), formula 2, to a compound of formula 2 and the carbonyl By reacting with a group reducing agent, the formula, ▲mathematical formula, chemical formula, table, etc.▼5-O-hydroxy protecting group-methyl-2,3- of formula 3 (in the formula, R is a hydroxy protecting group) (c) acylating or sulfonating or carbonylating the intermediate from step (b), formula 3, with a compound of formula 3 corresponding to group A; By reacting with the agent, formulas such as ▲mathematical formulas, chemical formulas, tables, etc. can be obtained.▼Formula 4 (In the formula, R is a hydroxy protecting group, and A is alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, arylthiocarbonyl, alkylsulfonyl , arylsulfonyl and carbonate groups, the alkyl moiety can be an unsubstituted or substituted C_1-C_3 alkyl group, the aryl moiety can be an unsubstituted or substituted phenyl group, and the alkyl moiety can be an unsubstituted or substituted phenyl group; O in which the substituents can be selected from 1 to 3 groups selected from halo and C_1 to C_3 alkoxy groups
1-O-activating group-5-O-hydroxy protecting group-2,
(d) converting the intermediate from step (c), formula 4, to a compound of formula 4 and a compound having one of formulas HX and TMSX; By reacting, there are formulas, ▲mathematical formulas, chemical formulas, tables, etc.▼Formula 5 (In the formula, R is a hydroxy protecting group, X is F, Cl,
1-Leaving group-5-O-protecting group-2,3-dideoxy-, which is a leaving group selected from Br and I
(e) converting the intermediate from step (d), formula 5, into a D- or -L-pentofuranose with an activated base selected from purine and pyrimidine bases, a base activated by reaction of pendant amino and hydroxy groups on the nucleus with an activating compound selected from silylating and acetylating and benzoylating agents, in the presence of one of a Brønsted acid and a Lewis acid, and a polar and (f) recovering the dideoxynucleotide from step (e). (2) the reducing agent used in steps (a) and (b) is B
H_3 (B_2H_6), BH_3・SMe_2, Na
BH_4, NaBH_4 and LiCl and AlCl_3
and one of BF_3, LiAlH_4, LiAlH(OMe)_3, and LiAlH(O-t-Bu)_3 and (Sia)_2
Claim (1) selected from dialkylboranes containing BH.
) method described. (3) The method of claim (1), wherein the reducing agent used in step (a) is BH_3 or BH_3SMe_2. (4) The hydroxy protecting groups used in step (a) include unsubstituted and substituted benzyl, trialkylsilyl, alkylarylsilyl, unsubstituted and substituted alkyl, vinyl,
A method according to claim (1), wherein the group is selected from benzoyl, mesitoyl, pivaloyl, unsubstituted and substituted atomoxy, and carbonate groups. (5) Claim (4) wherein the hydroxy protecting group is benzyl.
) method described. (6) A method according to claim 1, wherein the O-activating group used in step (c) is selected from alkylcarbonyl and arylcarbonyl. (7) A method according to claim 6, wherein the O-activated group used in step (c) is selected from acetyl and benzoyl. (8) the base component B in step (e) is activated by (i) the base reacting with a halotrialkylsilane as a silylating agent, where halo is selected from bromo and iodo; and (ii) 1- where the base has formula 5
Purines and pyrimidines selected from halopurine bases that are sufficiently reactive with the leaving group -5-O-protecting group -2',3'-dideoxypentofuranose and do not require activation of the base by reaction with an activating compound. selected from bases,
The method according to claim (1). (9) Base component B is selected from purine and pyrimidine bases activated by reaction with halotrimethylsilane, and the reaction in step (e) is CHCl_3, CH_2C
l_2, ClCH_2-CH_2Cl and CCl_4
Claim (8) wherein the process is carried out in a non-polar solvent selected from
) method described. (10) Base component B is 6-aminopurin-9-yl, 2
-aminopurin-9-yl, 2,6-diaminopurine-
the group of purine bases consisting of 9-yl, 2-amino-6-hydroxypurin-9-yl and 6-hydroxypurin-9-yl, and 2,6-dihydroxypyrimidine-
3-yl, 5-methyl-2,6-dihydroxypyrimidin-3-yl, 2-hydroxy-6-aminopyrimidin-3-yl, 5-vinyl-2,6-dihydroxypyridin-3-yl, 5-halovinyl -2,6-dihydroxypyrimidin-3-yl, 5-halomethyl-2,6-dihydroxypyrimidin-3-yl and 5-haloethyl-
The method according to claim 9, wherein the pyrimidine base is selected from the group of pyrimidine bases consisting of 2,6-dihydroxypyrimidin-3-yl. 11. The method of claim 1, comprising an additional step of removing the 5-O-hydroxy protecting group after step (e) and before step (f). (12) The 5-O-hydroxy protecting group is selected from benzyl and benzoyl groups, each of which is H_2/P
12. The process according to claim 11, wherein the removal is carried out by a method selected from catalytic hydrogenation of benzyl groups using a dCl_2 reagent and NH_3 hydrolysis of benzoyl groups. Formula (13), ▲Mathematical formula, chemical formula, table, etc.▼ (In the formula, R is a hydroxy protecting group, and A is an alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, arylthiocarbonyl, alkylsulfonyl, arylsulfonyl, and carbonate group. O selected from
- an activating group). (14) R is PhCH_2 (benzyl) and A is C
The compound according to claim (13), which is H_3CO (acetyl). Formula (15), ▲Mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R is a hydroxy protecting group, X is F, Cl,
Br and I). (16) The compound according to claim (15), wherein R is selected from PhCH_2 (benzyl) and benzoyl group. (17) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A compound with (18) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A compound with (19) A (L)-2',3'-dideoxynucleoside having the formula, ▲Mathematical formula, chemical formula, table, etc.▼ (wherein B is a member selected from the group consisting of purine and pyrimidine bases). (20) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (19), which has the following. (21) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (19), which has the following. (22) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (19), which has the following. (23) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (19), which has the following. (24) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (19), which has the following.