AU621615B2 - Creación de Inventos y Patentes con Inteligencia Artificial

AU621615B2 – Process for the preparation of dideoxycytidine
– Google Patents

AU621615B2 – Process for the preparation of dideoxycytidine
– Google Patents
Process for the preparation of dideoxycytidine

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Publication number
AU621615B2

AU621615B2
AU34686/89A
AU3468689A
AU621615B2
AU 621615 B2
AU621615 B2
AU 621615B2
AU 34686/89 A
AU34686/89 A
AU 34686/89A
AU 3468689 A
AU3468689 A
AU 3468689A
AU 621615 B2
AU621615 B2
AU 621615B2
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AU
Australia
Prior art keywords
bromide
dideoxycytidine
yield
mixture
amino
Prior art date
1988-05-12
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AU34686/89A
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AU3468689A
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Inventor
Peter Stefan Belica
Tai-Nang Huang
Percy Sarwood Manchand
John Joseph Partridge
Steve Tam
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F Hoffmann La Roche AG

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F Hoffmann La Roche AG
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1988-05-12
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1989-05-11
Publication date
1992-03-19

1989-05-11
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F Hoffmann La Roche AG

1989-11-16
Publication of AU3468689A
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patent/AU3468689A/en

1992-03-19
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1992-03-19
Publication of AU621615B2
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patent/AU621615B2/en

1993-01-28
Assigned to F. HOFFMANN-LA ROCHE AG
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F. HOFFMANN-LA ROCHE AG
Alteration of Name(s) in Register under S187
Assignors: F. HOFFMANN-LA ROCHE AG

2009-05-11
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Classifications

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07D—HETEROCYCLIC COMPOUNDS

C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom

C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings

C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS

C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof

C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen

C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom

C07H19/06—Pyrimidine radicals

Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE

Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

Y02P20/00—Technologies relating to chemical industry

Y02P20/50—Improvements relating to the production of bulk chemicals

Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Description

62 1 ef 689 FORM 10 COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 COMPLETE SPECIFICATION
(ORIGINAL)
FOR OFFICE USE: Class Int Class Complete Specification Lodged: Accepted: Published: *a Priority: Related Art:
I_
.o Name and Address of Applicant: Address for Service: F Hoffmann-La Roche Co Aktiengesellschaft Grenzacherstrasse 124-184 4002 Basle
SWITZERLAND
Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia r, a Complete Specification for the invention entitled: Process for the Preparation of Dideoxycytidine The following statement is a full description of this invention, including the best method of performing it known to me/us 5845/5 I RAN 4430/29 Abstract 21,3′-Dideoxycytidine is prepared from cytidine by a reaction sequence comprising the formation of novel cytidine derivatives.
0 C~ 0000 so 0 C 000 00000 00e Ga 00 0 000 S00 060 a 00000 0 0 0 00.000 0 0 rr. rr Lapl l~ll~ c-~ner~ IA The instant invention relates to a process for the preparation of 2′,3′-dideoxycytidine (ddC) and to novel intermediates used therein.
In accordance with the instant invention, 2′,3′-dideoxycytidine is prepared by a process comprising the steps of: protecting the 4-amino group of cytidine with a suitable protecting group, reacting the 4-amino protected cytidine with hydrogen bromide in acetic acid or with substituted or unsubstituted 2-acetoxy-2-methylpropanoyl bromide, 2-acetoxybenzoyl bromide, or 2-acetoxypropanoyl bromide to yield the corresponding 2′-bromo-3’5′-acylated or 3′-bromo-2′,5′ acylated 4-amino protected cytidines, reductively eliminating bromide and the 2′ or 3′-acyloxy groups from the product of step to yield the corresponding L 2′,3-didehydro derivative, hydrogenating the 2′,3′-didehydro derivative to yield 4 amino, S» 5′-hydroxy protected 2′,3′-dideoxycytidine.
removing the 4-amino and 5′-hydroxy protecting groups to yield o| :2′,3′-dideoxycytidine.
b The process comprising steps to is depicted in Scheme I 20 below: i 1 j 1459u 2 Scheme I .Hd OH T7 00 C0 C aC 00 C 0004 00 a 0 0 0 00 00 a 0 CC 000 0 08 0 Be- OAe
A
11-16 NHC P RoN
HC
I ,lo N
HO
Ii.
In Scheme I, Ac means acetyl. R’ is substituted or unsubstituted lower alkyl, aryl, or aralkyl with the substituents selected from halogen, alkyl, nitro, or alkoxy; and R is substituted or unsubstituted 2-acetoxy-2-methylpropanoyl, 2-acetoxypropanoyl, or 2-acetoxybenzoyl with the substituents selected from lower alkyl, aryl, or aralkyl.
Lower alkyl means a straight or branched chain hydrocarbon containing from 1 to 7 carbon atoms. Aryl means substituted or unsubstituted phenyl with the substituents selected from lower alkyl, lower alkoxy, nitro, amine, or halogen. Lower alkoxy means a lower alkyl ether group where the alkyl is as defined herein. Halogen means chlorine, fluorine, bromine, or iodine. Aralkyl means alkyl substitu- S 15 ted with one or more aryl groups.
C r CC The compounds of formula liIa, IIIb, IV and V are novel eI69 a and form part of the instant invention.
e€€t S 20 In step a) the 4-amino group of cytidine is protected with a suitable protecting group by methods known in the art. For example the reaction of cytidine with acetic tc C anhydride according to known methods yields N-acetyl Scr cytidine.
Step b) comprises a novel bromoacetylation step and yields the compounds of Formula III.
S* In one embodiment of step b) the amino-protected S t 30 cytidine is _reacted with substituted or unsubstituted 2-acetoxy-2-methylpropanoyl bromide, 2-acetoxybenzoyl bromide, or 2-acetoxypropanoyl bromide.
Preferred in this embodiment is where N-acetylcytidine is reacted with unsubstituted 2-acetoxy-2-methylpropanoyl bromide, or-2-acetoxypropanoyl bromide.
nr.armuurrr~l^Y»I~ 4 In another preferred embodiment of step b) N-acetylcytidine is treated with HBr in acetic acid. The bromoacetylation of N-acetylcytidine according to either of the above embodiments results in a mixture of bromoacetates in high yield.
Step c) comprises a novel reductive elimination step. In one embodiment of step the compound of formula IIIa or IIIb is reduced over a zinc-copper couple to yield the compound of formula IV. In another embodiment of step c) the compound of formula IV may be obtained by utilizing an electrolytic reduction.
Step d) comprises the hydrogenation of the compound of I c 15 formula IV. Preferably, the hydrogenation is carried out over a palladium on carbon catalyst with solvent mixtures containing THF, particularly methanol and THF.
e C ddC is then obtained in step e) by removing the 4-amino 20 and 5′ hydroxy protecting groups by base hydrolysis.
As used herein, the term zinc/copper couple means a S combination of zinc and copper prepared according to the 6 C method set forth in Example SA. THF means tetrahydrofuran.
MeOH means methanol. Regioisomer means positional isomers ge t, such that two positions on the compound are interchangeable. For example, compounds IIIa and IIIb are regioisomers.
0 a ca 30 The present invention will be described in connection with the following examples which are set forth for the purpose of illustration only.
Example 1 g of N-acetylcytidine and 50 ml 30% hydrogen bromide in acetic acid and 5 ml acetic anhydride was heated in an
C~
I
i [i oil bath at 500 for 18 hours. This reaction was cooled to room temperature and dissolved in 250 ml of methylene chloride and washed 2 times with 250 ml of 0.05 M potassium phosphate buffer pH7, and 2 times with 250 ml of saturated 5 aqueous sodium bicarbonate. The aqueous layers were washed with 250 ml of methylene chloride. The combined organic layers were dried over anhydrous sodium sulfate and evaporated to dryness to yield 6.1 g of a mixture of N-acetyl-3′-bromo-3′-deoxycytidine-2′,5′-diacetate and the regioisomer thereof.
Example 2 A suspension of 142.6 g (0.5 mole) of N-acetylcytidine, and 1.25 L of acetonitrile was stirred under nitrogen, cooled to o5C and treated dropwise with 225 ml of 2-acetoxy- -2-methylpropionyl bromide during 30 minutes. At the completion of the addition, a homogeneous solution resulted.
It was stirred at room temperature overnight (the reaction 20 was complete within 3 hr), cooled to 5 0 C, and diluted with 1.25 L of ethyl acetate. After recooling to 5 0 C, 2.0 L of saturated sodium bicarbonate was added. The mixture was stirred for 5 minutes, the organic phase was separated, and the aqueous phase was back-extracted with 500 ml of ethyl 25 acetate. The combined organic extracts were washed with 1 L of saturated brine, dried (MgSO 4 and evaporated to give a gum. Final drying at 40 0 C (1mm) for 1 hr gave 264.7 g of a white solid. High pressure liquid chromatographic analysis gave the following results (major peaks only): C Cr ‘cCC C C
C
CCp C e C C Relative Retention Time (RRT) [min] Compound 0
II
III(a) R=(CH 3 2
R’=CH
3 III(b)
OAC
48.77 60.20 6 6 HPLC conditions are methanol: water (40:60) with C 8 18 column and detection at 245 nm.
Example 3 28.52 g of N-acetylcytidine in 250 ml of acetonitrile was cooled to 10 0 C and treated with 48.75 g of (S)(-)-2-acetoxypropionyl bromide during 15 minutes. It was stirred at room temperature overnight, cooled to 100C, treated with 400 ml of cold (OOC) saturated sodium bicarbonate, and extracted with 250 ml of ethyl acetate. The extract was washed with 200 ml of saturated brine, dried (MgSO 4 and evaporated to give 45.45 g of a white foam.
1 o* 15 High pressure liquid chromatographic analysis gave the o a oa following results (major peaks only): 0 0 0 00 o. Relative Retention
E
000 Compound Time (RRT) [min] 0 00 .»oO 20 O III(a) R=CH 3 CH-C-, R’=CH 3 28.53 III(b) I 37.60 24 0oQ 0
OAC
0. 0 O 00 Reversed phase chromatography (C 18 column) with .00 V methanol in water gave a pure sample of III(a), N-acetyl-3′- -bromo-3′-deoxycytidine-2′-acetate-5′-[2-(acetoxy)propionate].
o0 Example 4 0
S
g of N-acetylcytidine in 45 ml of methylene chloride was cooled to 5 0 C and treated with 11.5 g of 2-acetoxy- -benzoyl bromide. Stirring was continued at room temperature for 4h, and the mixture was diluted with 70 ml of sodium bicarbonate. The organic phase was separated and the aqueous-phase was extracted with 50 ml of methylene chloride. The combined organic phase was washed with 7 saturated brine, dried (MgSO 4 and evaporated to give i 10.4 g of a foam. Chromatography of a 3.5 g portion by reversed phase chromatography (C 18 column) with methanol in water gave 12.2 g of a mixture of bromoacetates, N-acetyi-3′-bromo-3′-deoxycytidine-2′-acetate-5′-[2-(acetyloxy)benzoate] IIIa and its regioisomer IIIb, UV (EtOH):298 (E 9,600), 243 (E 20,000) and 217 (E 17,500) nm.
Relative Retention Compound Time (RRT) rminl III(a) R 2-acetoxybenzoyl 53.60 29.6 III(b) R’ CH 3 58.40 48.3 Example |j e A) 4.50 kg of zinc dust was washed with 3.75 L of 3% e aqueous hydrochloric acid by stirring for 3 to 5 minutes.
V t The hydrochloric acid was decanted from the solid. This o cycle was repeated with 3 x 3.75 L 14.0 L of 3% hydro- 20 chloric acid. The zinc dust was then washed with 4 x 3.0 L 12.0 L of deionized water to remove any residual hydrochloric acid. After all the water was decanted, the spongy 0oo0 a 0 zinc layer was treated with a solution made by dissolving t04 240.0 g of cupric sulfate dihydrate in 7.5 L of deionized water. The suspension was stirred rapidly as the solution -ao was added. The aquamarine color of the cupric sulfate solution was removed almost immediately and the zinc suspension changed in color from gray to black. The near colorless aqueous layer was decanted and the solid was S 30 washed with 4 x 3.0 L 12.0 L of deionized water. The suspension of zinc-copper couple was filtered then washed successively with 4 x 3.0 L 12.0 L of ethanol and 3 x L 9.0 L of ether. The blick solid was air-dried and then at 250 and 140 mm overnight and for 3 hr at 130-140 0
C
(0.5 mm) to remove ether. The solid was cooled to room temperature-under vacuum and was stored under argon in amber bottles. The procedure yielded 3.84 kg of zinc-copper couple.
8 B) 1.26 kg of N-acetyl-3′-bromo-3′-deoxycytidine-2′,5′- -diacetate and its 2′,3′-regioisomer which contained varying amounts of ethanol was dissolved in 4.0 L of acetonitrile.
The solution was evaporated to dryness at 500 and 70 mm to remove the residual ethanol. 1.26 kg of dry bromoacetate was dissolved in 16.0 L of acetonitrile. 350.0 g of zinc-copper couple was added and the heterogeneous mixture was degassed at 70 mm and layered with a nitrogen atmosphere. The mixture was degassed and relayered with nitrogen twice and a small volume of nitrogen was continuously bubbled into the mixture with a gas inlet tube that extended below the surface. HPLC analysis showed the reaction to be complete after stirring overnight (16-18 hr) at room temperature. The reaction mixture was filtered through a Celite pad and the filter bed I c 15 washed with acetonitrile. Then 250 ml of acetic anhydride, o and 250 ml of pyridine were added to reacetylate any 0 deacetylated product. The reaction mixture was immediately 0 et oe concentrated at 500 (70 mm). The residue obtained from two a runs was suspended in 40.0 L of methylene chloride and 0 20 washed with a warm solution of 1.50 kg of sodium bicarbonate and 1.50 kg of disodium ethylenediamine tetracetate (disodium EDTA) previously dissolved in 15.0 L of deionized 0 0 0 QC C water by heating on a steam bath to 50 6 C. After the layers Iwere separated, a fresh batch of 15.0 L of buffered EDTA solution was added to the aqueous layer and the aqueous Slayer extracted with 40.0 L of methylene chloride by stirring for 15 minutes. The organic layer was separated and the aqueous layer filtered to remove insoluble solids.
0 e These solids were suspended in 15.0 L of warm buffered EDTA 30 solution and extracted with 40.0 L of methylene chloride with rapid stirring for 15 minutes. All of the organic layers were washed in succession with 2 x 15.0 L 30.0 L of warm buffered EDTA and 20.0 L of saturated sodium bicarbonate solution. The layers were separated and the organic layers were collected in six 20 L glass bottles.
Each bottle-was dried over anhydrous sodium sulfate containing 20 g of SG extra charcoal with stirring. The 9 mixture was filtered through a 1″ Celite pad and concentrated at 50 0 (70 mm) to leave a dark semisolid. This solid was triturated with 4.0 L of cold tetrahydrofuran and 2.0 L of petroleum ether. The tan solid was dried to constant I 5 weight to give 986.0 g of N-acetyl 2′,3′-didehydro-2′,3′- -dideoxycytidine 5′-acetate. HPLC analysis showed this material to be 93% pure, but suitable for use in the next step.
Yields in other reactions ranged from 45-55%. An analytical sample was prepared by recrystallization from tetrahydrofuran to yield white solid, m.p. >3500; 15.80 (c 0.33, DMSO): lit. m.p. >2800 [T.
Adachi, T. Iwasaki, I. Inoue, and M. Miyoshi, J. Orq. Chem., 4, 1404 (1979)].
se C c Example 6 t’0 t 259.0 g of a mixture of bromoacetates from Example 2 in 20 2.5 L of acetonitrile was deoxygenated by evacuation followed by filling the reaction vessel with argon; this procedure was repeated three times. 100 g of zinc-copper couple (obtained as in Example 5A) was added, and the S mixture was stirred under argon at room temperature overnight. It was filtered over Celite, the flask was I rinsed out with 200 ml of acetonitrile, and the rinse was used to wash the Celite. The combined filtrate and washing were evaporated and the residue was dissolved in S 1.25 L of methylene chloride. This was added to a previously prepared solution of 200 g of ethylenediaminetetraacetic acid disodium salt dihydrate in 2.0 L of deionized water containing 200 g of sodium bicarbonate. The mixture was stirred vigorously for 1.5 hr, and filtered ovar Celite, which was washed with 300 ml of methylene chloride.
The organic phase was separated and the aqueous phase was re-extracted with 500 ml of methylene chloride. The combined organic phase was washed with 250 ml of saturated w I sodium bicarbonate, which was back-extracted with 100 ml of methylene chloride. The combined organic phase was dried (MgSO 4 filtered, and concentrated to ca. 800 ml. To this was added 30 ml of acetic anhydride followed by 40 g of poly-4-vinylpyridine and the mixture was stirred under nitrogen for 3 hr. It was filtered over Celite, which was washed with 200 ml of methylene chloride. The combined filtrate and washing were evaporated, 250 ml of toluene was added, and the mixture was evaporated again. 500 ml of ether was added with vigorous stirring for 15 minutes. The mixture was filtered and washed with 200 ml of ether to give 143.3 g of a mixture of N-acetyl-2′,3′-didehydro-2′,3′- -dideoxycytidine 5′-acetate and N-acetyl 2′,3′-didehydro- -2′,3′-dideoxycytidine 5′-[(2-acetoxy-2-methyl)propionate] as a tan-colored solid.
HPLC Analysis Relative Retention Compound Time (RRT) [min] IV, R=CH 3
R’=CH
3 5.83 6 VI, R=(CH 3 2
R’=CH
3 C 13.50 91
OAC
Recrystallization of the mixture from hot tetrahydrofuran yielded pure N-acetyl 2′ 3′-didehydro-2′ 3′-dideoxy- C t C I C I cytidine 5′-[(2-acetoxy-2-methyl)propionate], mp 173-1750; [a]D 123.50 (c 1.0, CHC1 3 Example 7 A total of 1.47 g of a mixture of bromoacetates from Example 3 was reduced with 800 mg of zinc-copper couple as described in Example 6 to give 570 mg of N-acetyl-2′,3′- -didehydro-2′,3′-dideoxycytidine 5′-[(2-acetoxy)propionate] after crystallization from hot tetrahydrofuran, mp 125 0
C;
[a]D 119.04 (c 0.25, CHC13).
I Example 8 30.5 g of a mixture of the bromoacetates of Example 4 was deoxygenated by evacuation followed by filling the reaction vessel with argon; this procedure was repeated twice. 10 g of zinc-copper couple (obtained as in Example was added and the mixture was stirred under argon at room temperature overnight. It was filtered over Celite, and the flask was rinsed out with 20 ml of acetonitrile, and the rinse was used to wash the Celite. The combined filtrate and washings were evaporated, and the residue was dissolved in 125 ml of methylene chloride. This was added to a previously prepared solution of 20 g of ethylenediaminetetracetic acid disodium salt dihydrate in 200 ml of water containing 20.0 g of sodium bicarbonate. The mixture v was stirred at room temperature for 1.5 h, and filtered over Celite. The organic phase was separated and the aqueous ‘phase was re-extracted with 75 ml of methylene chloride.
The combined organic phase was dried (MgSO 4 and 20 filtered. The filtrate was treated with 6.0 ml of acetic anhydride and 850 mg of 4-dimethylaminopyridine. The mixture was stirred at room temperature for 2 h, diluted with 25 ml of ethanol, and then concentrated to ca. 100 ml, S’ 25 ml of toluene was added and the mixture was evaporated to dryness. The residue was crystallized from 100 ml of ether-ethanol to give 6.3 g of N-acetyl-2,’3’didehydro-2′,3′-dideoxycytidine 5′-[2(acetoxy)benzoate].
Recrystallization from hot ethanol, tetrahydrofuran (1:1) .ttO, gave an analytical sample of mp 265 0 C UV (EtOH): 299 30 (E 9,800), 243 (E 21,550), and 203 (E 48,400) nm.
t t Example 9 To the catholyte reservoir of an electrolytic cell was added 35.0 g of N-acetyl-3′-bromo-3′-deoxycytidine and 1.0 L of 0.25M tetraethylammonium tosylate in acetonitrile. To the anolyte reservoir was i I; -i l–I~E4~SYIP~I 12 added 1.0 L of .025 M tetraethylammonium tosylate in acetonitrile. Both the catholyte and anolyte were circulated through the electrolytic cell at a flow rate of 200 ml/min/ cell. The cell was divided by an anion exchange membrane and the initial current density was 2.4 mA/cm at -1.5V. The reaction was followed by TLC and HPLC. During the first 8 hr the desired product N-acetyl 2′,3′-dideoxycytidine precipitated from the reaction mixture. The product was removed by filtration and the reaction continued. After 16 hr, this reaction was almost complete.
The catholyte mixture was collected and evaporated to dryness at room temperature and 10 mm vacuum. The dried residue was combined with the previously filtered solids.
The mixture of product and electrolyte was then dissolved in 200 ml of deionized water. The mixture was extracted with 3 x 300 ml 900 of methylene chloride. The organic extract was dried over anhydrous sodium sulfate and evaporated to a o o .00 dryness. The residue was triturated with 180 ml of oo o o 90 tetrahydrofuran to give 18.12 g (76% yield) of N-acetyl 20 2′,3′-didehydro-2′,3′-dideoxycytidine o o (IV, R=CH3CO. R’=CH3), m.p. >3000 (dec); [a]D 14.70 (c 0.387, DMSO). The product was analyzed by HPLC to give a 99.2% assay.
0 0* 25 Example 0 00 0 0 0 000 0 To the catholyte reservoir of an electrolytic cell was added 10.0 g (0.019 mol) of N-acetyl-3′-bromo-3′-deoxy- A 00 00: cytidine 5′-[(2-acetoxy-2-methyl)propionate] and 500 ml of 30 0.25 M tetraethylammonium tosylate in acetonitrile. To the anolyte reservoir was added 500 ml of .025 M tetraethylammonium tosylate in acetonitrile. Both the catholyte and anolyte were circulated through the electrolytic cell at a flow rate of 250 ml/min/cell. The cell was divided by an anion exchange membrane (lonac MA-3475, Sybron Chemical Division, Birmingham, NJ) and the initial current density a 2 was 0.8 mA/cm at -4.0 V. The reaction was monitored by -13- TLC and HPLC. During the first 30 minutes, a sample analyzed gave 60% conversion. About 90 minutes later, the TLC sample showed total disappearance of the starting material. The catholyte solution was collected and evaporated to dryness at room temperature in vacuo. The dried residue was then dissolved in 200 ml of deionized water. The solution was extracted with 3 x 200 ml 600 of methylene chloride. The organic extract was dried over anhydrous sodium sulfate and was then evaporated to dryness to yield 3.07 g of tan-colored solids (yield, A total of 0.5 g of the product was dissolved in 8 ml of hot tetrahydrofuran and allowed to stand overnight to effect crystallization. The solid crystalline mass was filtered and washed with 10 ml of ether to give .22 g of N-acetyl 2′,3′-didehydro-2′,3′-dideoxycytidine 5′-[(2-acetoxy-2- CC -methyl)propionate]. Recrystallization from hot tetrahydro- S furan gave crystals m.p. 170-172 0
C.
a c 0 C S, Example 11 S* To 16.0 L of tetrahydrofuran and 22.0 L of methanol, a slurry of 500.0 g of N-acetyl 2′,3′-didehydro-2′,3′- -dideoxycytidine 5′-acetate in 1.5 L of methanol was given.
Then any solid starting material was washed from the S 25 glassware into the flask with an additional 500 ml of e cc methanol. With the mixture layered with argon, 20.0 g of palladium on carbon catalyst in 200 ml of methanol was carefully added. The flask was then evacuated (70 mm) and layered with hydrogen gas. The evacuation-layering process r 30 was carried put three times. A hydrogen pressure of 1 t«C C atmosphere was then maintained as the mixture was stirred at room temperature. After 52 minutes the reaction slowed markedly and a sample was removed for HPLC analysis to trace the disappearance of starting material. The flask was evacuated (70 mm) and layered with argon gas. This evacuation-layering process was carried out three times.
The reaction mixture was then cautiously filtered through a 14 Celite pad. The pad was then washed with 500 ml of ca. tetrahydrofuran in methanol. The filtrate was then concentrated to dryness at 500 (70 mm) on a large rotary evaporator. In the same manner 9 batches were hydrogenated to a mixture of N-acetyl 2′,3′-dideoxycytidine and a small amount of N-acetylcytosine by-product. The filtrate was concentrated at 70-800 (70 mm) on a large rotary evaporator or give a white to off-white solid. The solid was triturated with 4.5 L of acetonitrile, by stirring the solid for 15 minutes. The paste was cooled to 100 for minutes and filtered to remove acetonitrile soluble impurities [small amounts of overhydrogenated products such as 5,6-dihydro-2′,3′-dideoxyuridine 5′-acetate]. The white filter cake was then dissolved in 20 L of hot acetonitrile to give a cloudy solution. This solution was filtered through a pad of Celite to remove insoluble N-acetylt t’ cytosine. The colorless filtrate was concentrated at 70-800 S (70 mm) to a 10 L volume. The concentrated solution was chilled to 100 for 1 hr to induce crystallization. The 20 solid was collected on a Bichner funnel and washed with 2 x 2.0 4.0 L of cold acetonitrile. The white solid was dried at 800 to yield 3.30 kg of first crop N-acetyl 3-dideoxycytidine 5′-acetate, m.p. 210-2110 25 t [Ca] =+92.00 (c 0.49, CH 3
OH).
S a D 3
CC
The mother liquors and washes were pooled from several oo 0 runs and concentrated. Recrystallization of this mterial yielded an additional 10-16% of N-acetyl 2′,3′-dideoxycytidine 5′-acetate of comparable quality. The mother 30 liquors from this second crop material contained small c C amounts of the 5,6-dihydro-2′,3′-dideoxyuridine Purification of these mother liquors on silica gel with 10:1 ethyl acetate-methanol gave an additional 3-5% of desired product.
Example 12 142.3 g of the bromoacetates from Example 5B) in 800 ml of methanol was warmed until a solution was obtained, diluted with 800 ml of tetrahydrofuran, and then cooled to room temperature. A total of 8.9 g of 10% palladium on charcoal was added under argon and the mixture was hydrogenated with stirring at room temperature and atmospheric pressure until hydrogen uptake ceased (11 L, ca.
3 hr). The mixture was filtered over Celite and the Celite was washed with 300 ml of methanol. The combined filtrate and washing were evaporated to give 132.7 g of a mixture of N-acetyl-2′,3′-dideoxycytidine 5′-acetate and N-acetyl- -2′,3′-dideoxycytidine-5′-[(2-acetoxy-2-methyl)propionatej.
HPLC Analysis
I
«e Relative Retention o1 Compound Time (RRT) Cmin 1. 0, 20 V R=CH 3
R’=CH
3 C 7.40 6 V R=(CH 3 2 R’=CH 3
C
OAC 17.93 SN-Acetylcytosine 3.67 1 A pure sample of N-acetyl-2′,3′-dideoxycytidine -acetyl-2-methyl)propionate] was obtained as a foam by chromatography over silica gel with 1 percent methanol in 25 methylene chloride, [l]D 136.08 (CHCl 3 C=1.02.
0 C 30 Example 13 a t A solution of 720 mg of the product from Example 7 in ml of methanol and 10 ml of tetrahydrofuran was hydrogenated over 200 mg of 10% palladium on charcoal at room temperature and atmospheric pressure until hydrogen uptake ceased (10 ml). The mixture was filtered over Celite and the filtrate was evaporated to give a gum. Chromato- 16 16 graphy on 10 g of silica with 10% methanol in methylene chloride, gave 290 mg of N-acetyl-2′,3′-dideoxycytidine 5′-[(2-acetoxy-2-methyl)propionatej as a foam, [a] 88.43 (C 0.99, CHC13); UV (EtOH): 299 (E 6,420), 246 (E 12410), and 214 (E 16,500, nm).
Example 14 A solution of 20.7 g of the product from Example 13 in 100 ml of ethanol was treated with 10.0 ml of Triton B (N-benzyltrimethyl-ammonium hydroxide), and the mixture was stirred at room temperature overnight. The mixture was concentrated to 20 ml, cooled to 0 C, and the product was collected by filtration. It was washed with 10 ml of cold ethanol to give 4.48 g of 2′,3′-dideoxycytidine, mp 215-218 0 C, as an off-white solid.
e 2 C 9. Example o040 oo a oe a 20 To 1.66 g of the product of Example 7 in 30 ml of methanol and 30 ml of dimethylformamide, 300 mg of palladium on charcoal was added under argon and the mixture was hydrogenated with stirring at Loom temperature and Sort atmospheric pressure until hydrogen uptake ceased (113 ml).
The mixture was filtered over Celite and the filtrate was evaporated in vacuo to give a gum, which was chromatographed 00o 6 on 15 g of silica gel with 2% methanol in methylene chloride as eluent to give 970 mg of N-acetyl-2′,3′-dideoxycytidine 00..06 5′-[2-(acetoxy)benzoate] as a white foam; UV (EtOH); 298 S30 (E 9,450) 243 (E 19,500), and 203 (E 42,350) nm.
C C Example 16 A solution of 33.4 g of the product of Example 15 in 330 ml of methanol, 33 ~il. of water, and 66 ml of triethylamine was stirred, under argon, at 65 0 C for 7h, and Sthen at room temperature overnight. It was concentrated in 17 vacuo and the residue was azeotroped three times with toluene. The residue was dissolved in 50 ml of ethanol, and diluted with 400 ml of acetone, and stirred at room temperature overnight. The product was collected by filtration to give 4.04 g of 2′,3′-dideoxycytidine, m.p.
218-220 0
C.
Example 17 1.55 kg of N-acetyl 2′,3′-dideoxycytidine 10.6 L of methanol, 3.1 L of triethylamine, and 1.56 L of deionized water was heated to 60-650 (bath temperature). The mixture was stirred at 55-600 for 3 hr, then at ambient temperature for 12 h. The reaction mixture was concentrat i S: 15 in vacuo at 70-800 (70 mm) to a volume of 2.5 L to induce cot crystallization. To the white semi-solid was added 1.5 L of ,C absolute ethanol and again the mixture was concentrated in S, vacuo to 2.5 L volume to remove residual solvents. The white semi-solid was cooled to 10 0 C for 0.5 hr and was collected by filtration and washed with 1.5 L of absolute ethanol. The damp white solid was recrystallized from 50.0 L of absolute ethanol at reflux and filtered to remove S» any particulate matter. The filtrate was concentrated in vacuo at 70-80° (70 mm) to a volume of 3.0 L. The semi-solid was then heated for 10 minutes without vacuum at reflux in C C the rotary evaporator. The mixture was cooled to 10 0 C for c 2 hr to effect crystallization and the white solid was collected by filtration, and washed with 2 x 1.0 L 2.0 L t of absolute ethanol. The solid was dried in a vacuum oven SCfcr 30 at 85 0 C andl mm overnight to yield 1.02 kg of 2′,3′-dideoxycytidine as a white solid, m.p. 225-228 0
C;
[a]D 76.90 (c 0.56, H 2 2 105.00 (c 0.50, CH OH); lit. m.p. 215-2170; [a]25 3 D 810 (c 0.635, H 0) Horowitz. J. Chua, M. Noel, and J. T. Donatti, J. Orq. Chem., 32, 817 (1967)].
18 The mother liquors from three batches were pooled and concentrated to a volume of 1.0 L to effect recrystalli.
zation. This solid was recrystallized from 5.0 L of absolute ethanol which was concentrated to a volume of 1.0 L to yield 2-4% additional 2′,3′-dideoxycytidine.
Example 18 27.2 g of the mixture of the compounds from Example 12 in 71.0 ml of methanol was stirred at room temperature until a solution was obtained and then treated with 7.14 ml of Triton B (40% Benzyltrimethyl-ammonium hydroxide in methanol). Stirring was continued at room temperature overnight and the product was collected by filtration. It was washed with some cold methanol to give 8.33 g of crude 2′,3′-dideoxycytidine, with a purity of 99.17% (HPLC).
o Evaporation of the filtrate and washing gave a semisolid to F ,which 20.0 ml of ethanol was added. The product was cte collected by filtration, washed with some cold ethanol, to 20 give an additional 1.05 g of crude (96.67% by HPLC) t 2′,3′-didexoycytidine, a total of 9.38 g of crude 2′,3′-didexoycytidine (43.1% from N-acetylcytidine).
tC The combined crude 2′,3′-didexoycytidine above (9.38 g) was dissolved in a mixture of 100 ml of hot (reflux) Sabsolute ethanol and 12 ml of deionized water. The hot solution was filtered, and the funnel was washed with 10 ml of ethanol. The combined filtrate and washing were allowed .to cool to room temperature and cooled further to ca. 7 0
C
i; 30 (ice-bath). The product was collected by filtration and washed with a few ml of ethanol to give 7.17 g of 2′,3′-didexoycytidine. as white crystals, m.p. 219-221 0
C,
95.860 (CH30H, c=1.46).

Claims (7)

1. A process for the preparation of 2′,3′-dideoxycytidine comprising the steps of: protecting the 4-amino group of cytidine with a suitable protecting group, reacting the 4-amino protected cytidine with hydrogen bromide in acetic acid or with substituted or unsubstituted 2-acetoxy-2-methyl- propanoyl bromide, 2-acetoxybenzoyl bromide, or 2-acetoxypropanoyl bromide to yield the corresponding 2′-bromo-3’5′-acylated or 3′-bromo-2′,5′ acylated 4-amino protected cytidines, reductively eliminating bromide and the 2′ or 3′-acyloxy groups from the product of step to yield the corresponding 2′,3-didehydro derivative, hydrogenating the 2′,3′-didehydro derivative to yield 4 amino, 15 5′-hydroxy protected 2′,3′-dideoxycytidine. removing the 4-amino and 5′-hydroxy protecting groups to yield 2′,3′-dideoxycytidine.

2. The process of Claim 1 wherein the bromoacetylation is conducted with substituted or unsubstituted 2-acetoxy-2-methylpropanoyl 20 bromide, 2-acetoxybenzoyl bromide, or 2-acetoxypropanoyl bromide with the substituents selected from lower alkyl, aryl, or aralkyl; or with hydrogen bromide in acetic acid.

3. The process of Claim 1 wherein the bromoacetylation is conducted with 30 percent hydrogen bromide in acetic acid. 25

4. The process of any one of Claims 1 to 3 wherein the hydrogenation is conducted with a palladium carbon catalyst and tetrahydrofuran in methanol.

The process of any one of Claims 1 to 4 wherein bromine and the acetoxy group is eliminated from the bromoacetate derivative to yield the corresponding 2′,3′-didehydro derivative by electrolytic reduction.

6. 2′,3′-Dideoxycytidine, whenever prepared by the process of any one of claims 1 to 5 or by an obvious chemical equivalent thereof. 0000 I t~o0 00,. 0 r f 4 o 0 1459u 20

7. A process for the preparation of 2′,3′-dideoxycytidine substantially as hereinbefore described with reference to any one of the Examples. DATED this EIGHTH day of JANUARY 1992 F Hoffmann-La Roche Co Aktiengesellschaft Patent Attorneys for the Applicant SPRUSON FERGUSON 0 0 0 0 0 oa a oioo 00 0 oa o o a o0 o0000 o 00 0 o o 0 00 0 QO o 0 o a o 0 0 0 0a0 I Ti™MS/1459u c 1

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Cited By (1)

* Cited by examiner, † Cited by third party

Publication number
Priority date
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AU640860B2
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Substituted 2′, 3′-dideoxy-5-trifluoromethyluridines, processes for their preparation and their use in medicaments

Families Citing this family (25)

* Cited by examiner, † Cited by third party

Publication number
Priority date
Publication date
Assignee
Title

US5290927A
(en)

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1994-03-01
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(en)

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(en)

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2003-11-04
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US5204466A
(en)

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1993-04-20
Emory University
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US5276151A
(en)

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(en)

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AU7558491A
(en)

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1991-10-30
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Nucleoside derivatives

JPH0592970A
(en)

1991-03-01
1993-04-16
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US5817667A
(en)

1991-04-17
1998-10-06
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Compounds and methods for the treatment of cancer

US5192764A
(en)

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1993-03-09
Research Foundation Of State Of N.Y.
Pyrazinone n-oxide nucleosides and analogs thereof

WO1993006119A1
(en)

1991-09-27
1993-04-01
The Institute Of Cancer Research
Deoxynucleoside derivatives

RU2085557C1
(en)

1991-09-30
1997-07-27
Санкио Компани Лимитед
Pyrimidine nucleoside derivatives or their pharmaceutically acceptable salts and method of their synthesis

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(en)

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US20020120130A1
(en)

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2002-08-29
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US5587362A
(en)

1994-01-28
1996-12-24
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L-nucleosides

IL115156A
(en)

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2000-07-16
Univ Georgia
Pharmaceutical compositions for the treatment of cancer comprising 1-(2-hydroxymethyl-1,3-dioxolan-4-yl) cytosines

US6391859B1
(en)

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US5703058A
(en)

1995-01-27
1997-12-30
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Compositions containing 5-fluoro-2′,3′-didehydro-2′,3′-dideoxycytidine or a mono-, di-, or triphosphate thereof and a second antiviral agent

US5808040A
(en)

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1998-09-15
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L-nucleosides incorporated into polymeric structure for stabilization of oligonucleotides

AU722214B2
(en)

1995-06-07
2000-07-27
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Nucleosides with anti-hepatitis B virus activity

US5753789A
(en)

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NZ540956A
(en)

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(en)

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WO2013036846A2
(en)

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N4 derivatives of deoxycytidine prodrugs

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US4604382A
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