EP0640091A1 - Synthese de monomeres nucleotidiques - Google Patents

Synthese de monomeres nucleotidiques

Info

Publication number
EP0640091A1
EP0640091A1 EP93911072A EP93911072A EP0640091A1 EP 0640091 A1 EP0640091 A1 EP 0640091A1 EP 93911072 A EP93911072 A EP 93911072A EP 93911072 A EP93911072 A EP 93911072A EP 0640091 A1 EP0640091 A1 EP 0640091A1
Authority
EP
European Patent Office
Prior art keywords
synthesis
deoxyadenosine
contacting
benzoyl
yield
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP93911072A
Other languages
German (de)
English (en)
Other versions
EP0640091A4 (fr
Inventor
Leonid Beigelman
Jasenka Matulic-Academic
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sirna Therapeutics Inc
Original Assignee
Ribozyme Pharmaceuticals Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US07/882,479 external-priority patent/US5506349A/en
Application filed by Ribozyme Pharmaceuticals Inc filed Critical Ribozyme Pharmaceuticals Inc
Publication of EP0640091A1 publication Critical patent/EP0640091A1/fr
Publication of EP0640091A4 publication Critical patent/EP0640091A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • 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
    • 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
    • C07H19/16—Purine radicals

Definitions

  • This invention relates to the chemical synthesis of 2 ' ,3'-dideoxycytidine, 2' ,3'-dideoxyguanosine,
  • 2' ,3'-dideoxypurine-nucleosides i.e.. adenosine, guanosine and inosine nucleosides
  • 3'-0-benzoyl-2'- deoxyadenosine i.e. adenosine, guanosine and inosine nucleosides
  • this invention features an improved economical synthetic method for the preparation of dideoxyuridine (dd ⁇ ) which is then converted into dideoxycytidine (ddC) via a 4-triazolyl intermediate.
  • the method is not only cost efficient, but can be scaled up to several hundred gram quantities.
  • the method generally utilizes inexpensive uridine as a starting material which is converted in a 7 step reaction sequence to ddC with a yield of about 30%.
  • the ddC can be used for chemical synthesis of sugar-modified nucleotides; chemical synthesis of DNA chain terminators; and chemical synthesis of anti-HIV 2' ,3'-dideoxynucleosides.
  • the invention features a method for chemical synthesis of 2',3'- dideoxycytidine in which 5'-silylated-2',3'-deoxyuridine is used to form ddC, e.g.. by reaction with 1,2,4- triazole, a phosphorous oxychloride, followed by treatment with ammonia, and the silyl group of the resulting product cleaved by use of an ion-exchange resin (specifically, that referred to as the Amberlyst A-26 (F ⁇ )) in toluene.
  • an ion-exchange resin specifically, that referred to as the Amberlyst A-26 (F ⁇ )
  • the invention features a method for synthesis of dideoxyguanosine by directly converting N 2 -isobutyrylguanosine to a 2',3'-unsaturated derivative by action of acetoxyisobutyryl bromide in moist acetonitrile, with subsequent reductive elimination by Zn/Cu/DMF and deacylation by NaOMe/methanol.
  • Catalytic hydrogenation and ammonolysis of the N 2 -isobutyryl-2' ,3'- didehydroguanosine provides 2' ,3'-dideoxyguanosine in 30% yield.
  • the method of this invention utilizes N 2 - isobutyrylguanosine as a starting compound, and thus overcomes problems with isolation of unsaturated compounds.
  • the described method is cost efficient and can be scaled up to multi—gram quantity.
  • the invention features a method for chemical synthesis of 2' ,3'-dideoxyguanosine, including contacting N 2 -isobutyrylguanosine with acetoxyisobutyryl bromide in acetonitrile with subsequent reductive elimination, deacylation, hydrogenation and ammonolysis.
  • the invention features a convenient "one-pot" (i.e..
  • the method of this invention utilizes selective protection of the 5'-OH group of 2'-deoxyadenosine (or 2 '— deoxyinosine) and N 2 -isobutyry1-2'-deoxyguanosine by introducing a tert-butyldiphenylsilyl protecting group.
  • the high selectivity of this step allows the preparation of the above key compounds in a "one-pot" procedure without isolation of intermediates.
  • Simultaneous removal of the 5'-0-tert-butyldiphenylsilyl group during the elimination step reduces the total number of steps and gives a high yield of 2 r , 3'-unsaturated nucleosides.
  • the described methods are cost efficient, and can be scaled up to multi-gram quantity.
  • the invention features a method for chemical synthesis of 2',3'-dideoxyadenosine, 2',3'-dideoxyinosine, and 2' ,3'-dideoxyguanosine without purification of intermediate compounds by sequentially treating: (a) N 2 -isobutyry1-2'-deoxyguanosine with a t- butyl-diphenylsilylchloride, triphenylphosphine, diethyl- azodicarboxylate, p-nitrophenylethanol andmethanesulfonyl (or p-toluenesulfonyl) chloride; or (b) by treating.2'- deoxyadenosine or 2'-deoxyinosine sequentially with t- butyldiphenylsilylchloride, and methanesulfonyl (or p- toluenesulfonyl) chloride.
  • the invention features a method for synthesis of 3'-O-benzoyl-2'-deoxyadenosine based on the efficient protection of the 5'-hydroxyl group of 2'-deoxyadenosine, using stable t-butyldimethylsilyl ether (TBDMS) protection.
  • TDMMS stable t-butyldimethylsilyl ether
  • Acylation with benzoyl cyanide occurs exclusively at the remaining free sugar hydroxyl group.
  • Other benzoylating agents e.g. , benzoic anhydride, benzoyl chloride
  • the three step procedure of this invention provides high yields of 5'-0 and 3'-O-protection reactions.
  • the procedures used are simple and require no chromatographic purifications.
  • this three step synthesis is a useful improvement over existing methods and provides a high overall yield of about 59%.
  • the method can be scaled up for synthesis of kilogram quantities of the desired compounds.
  • the invention features a method for rapid and selective synthesis of 3'-0-benzoyl- 2'-deoxyadenosine, a protected nucleoside intermediate suitable for modification at C-5' and/or N-6 position.
  • the nucleoside is treated with a chemical which provides a protecting group at the 5'-OH group (e.g.. TBDMS) , and the remaining 3'-OH group is benzoylated using benzoyl cyanide.
  • the protecting group is then removed by treatment with a reagent such as tetrabutylammonium fluoride (TBAF) or its equivalent.
  • Fig. 1 is a diagrammatic representation of the chemical synthesis of ddC from uridine by a method of this invention.
  • Fig. 2 is a diagrammatic representation of the synthesis of 2' ,3'-dideoxyguanosine.
  • Fig. 3 is a diagrammatic representation of a method of this invention for synthesis of 2',3'- - dideoxyguanosine.
  • Fig. 4 is a diagrammatic representation of a method of this invention for synthesis of 2',3'- dideoxyadenosine and 2' ,3'-dideoxyinosine.
  • Fig. 5 is a diagrammatic representation of a method for synthesis of 3'-O-Benzoy1-2'-deoxyadenosine. Synthesis:
  • Example 1 2' ,3'-Dideoxycytidine
  • 2'-deoxy ⁇ ridine is formed from uridine by a method similar to that of Haoqiang and Chu, supra, and then converted to ddC by the steps shown.
  • uridine 60 g was treated with acetyl bromide (with or without HBr) in acetonitrile to give 3',5'-di-0-acetyl-2'-bromo-2'-deoxyuridine.
  • the crude material was deoxygenated using tributyltin hydride in the presence of benzoyl peroxide (or 2,2'-azobis-(2- methyl)propionitrile r or azobisisobutyronitrile) to give, after deacetylation with methanolic ammonia, 2'- deoxyuridine in 70% yield.
  • TBDMS t- butyldimethylsilyl
  • N 2 -isobutyryl-2' ,3'-dideoxy- 2' ,3'-didehydroguanosine was formed as follows.
  • N 2 -isobutyrylguanosine 6,74 g (20 mM) was suspended in 80 ml of dry acetonitrile. 0.4 ml water was added followed by acetoxyisobutyryl bromide and the mixture stirred by room temperature (about 24°C) for 1 hour.
  • the reaction mixture was filtered, diluted with 300 ml of ethylacetate and added to cold saturated NaHC0 3 solution. The organic layer was separated and washed by water (70 ml x 2) . The combined water extracts were washed by ethylacetate (2 x 250 ml) .
  • 2' ,3'-dideoxyguanosine was formed as follows: 2.6 g of N 2 -isobutyryl-2',3'-dideoxy-2',3'- didehydroguanosine (8.125 mM) in methanol (250 mml) was hydrogenated at 32 psi in the presence of 10% Pd/C (1.3 g) for five hours. The catalyst was filtered and the solvent evaporated. The purification of the crude product by flash chromatography using CHC1 3 methanol (10:1) and subsequent ammonolysis yielded 2.0 g (77%) of 2',3'- dideoxyguanosine as a solid.
  • Example 3 2' .3 '-Dideoxyguanosine Referring to Fig.
  • N 2 -isobutyryl-6-0-[ (p-nitrophenyl)ethyl]-3'-O-mesyl- 5'-O-tert-butyldiphenylsily1-2'-deoxyguanosine was obtained as a solid in 80% yield.
  • N 2 -isobutyryl-2 ' , 3 '-dideoxy-2 ' , 3 ' - didehydroguanosine was prepared as follows.
  • N 2 -isobutyry1-6-0-[ (p-nitrophenyl)ethyl]-3'-o- mesyl-5'-0-tert-butyldiphenylsilyl-2'-deoxyguanosine 4.77 g (6 mM) was dissolved in 60 ml of dry N,N- dimethylformamide and 1.62 g (30 mM, 5 eq) sodium methanolate was added. After 1 hour 50 ml methanol was added, and the reaction mixture neutralized by Amberlite IRC 50 (H + ) .
  • N 2 -isobutyryl-2' ,3'-dideoxyguanosine was prepared as follows.
  • dideoxyadenosine and dideoxyinosine can be formed in a similar manner with similar yields to that described in Example 3.
  • reaction mixture was kept for 2 hours at 0°C, then 24 hours at room temperature (about 24°C) , with exclusion of moisture, and 30 ml methanol added. After an additional 30 minutes, the reaction mixture was evaporated to dryness and coevaporated with toluene. The residue was dissolved in chloroform (500 ml) , washed with water (3 x 100 ml) , dried (MgS0 4 ) , evaporated and purified by flash chromatography on silica gel (800 g) in CHC1 3 - 2% MeOH. 3',O-tosyl-5'-0-t- butyldiphenylsilyl-2'-deoxyadenosine was obtained as a solid in 85% yield. 2',3'-dideoxy-2' ,3'-didehydroadenosine was prepared as follows:
  • the invention generally features a three step procedure which can be performed without chromatographic purification steps.
  • Precipitate was formed, and after 1 hour of stirring at room temperature, water and chloroform were added and the mixture filtered. The precipitate was washed successively with chloroform and water and dried to give 5'-0-TBDMS-2'- deoxyadenosine in 85% yield.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Saccharide Compounds (AREA)

Abstract

L'invention se rapporte à des procédés de synthèse de 2',3'-didésoxycytidine, 2',3'-didésoxyguanosine, 2',3'-didésoxyadénosine, 2',3'-didésoxyinosine et 2',3'-didésoxyguanosine, et de 3'-O-benzoyl-2'-désoxyribonucléoside.
EP93911072A 1992-05-13 1993-05-06 Synthese de monomeres nucleotidiques. Withdrawn EP0640091A4 (fr)

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
US919544 1978-06-27
US882572 1986-07-07
US88257292A 1992-05-13 1992-05-13
US07/882,479 US5506349A (en) 1992-05-13 1992-05-13 Chemical synthesis of 2', 3'-dideoxycytidine
US882479 1992-05-13
US91064992A 1992-07-08 1992-07-08
US910649 1992-07-08
US91954492A 1992-07-24 1992-07-24
PCT/US1993/004239 WO1993023413A1 (fr) 1992-05-13 1993-05-06 Synthese de monomeres nucleotidiques

Publications (2)

Publication Number Publication Date
EP0640091A1 true EP0640091A1 (fr) 1995-03-01
EP0640091A4 EP0640091A4 (fr) 1995-07-26

Family

ID=27505972

Family Applications (1)

Application Number Title Priority Date Filing Date
EP93911072A Withdrawn EP0640091A4 (fr) 1992-05-13 1993-05-06 Synthese de monomeres nucleotidiques.

Country Status (6)

Country Link
EP (1) EP0640091A4 (fr)
JP (1) JPH07508022A (fr)
AU (1) AU4234193A (fr)
CA (1) CA2135498A1 (fr)
MX (1) MX9302761A (fr)
WO (1) WO1993023413A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW374087B (en) * 1993-05-25 1999-11-11 Univ Yale L-2',3'-dideoxy nucleotide analogs as anti-hepatitis B(HBV) and anti-HIV agents
US5627160A (en) * 1993-05-25 1997-05-06 Yale University L-2',3'-dideoxy nucleoside analogs as anti-hepatitis B (HBV) and anti-HIV agents
EP1910393A2 (fr) * 2005-07-05 2008-04-16 Hetero Drugs Limited Nouveau procede de preparation de didanosine au moyen de nouveaux produits intermediaires
CN105884846B (zh) * 2016-06-01 2018-08-28 中南大学 一种2’-脱氧腺苷的合成方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US64631A (en) * 1867-05-14 Thomas g
JPS63275597A (ja) * 1987-05-07 1988-11-14 Ajinomoto Co Inc ジデオキシシチジンの製造方法、その製造中間体、およびその製造方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5099010A (en) * 1983-01-17 1992-03-24 Research Corporation Intermediates in the preparation of 3'-amino-2',3'-dideoxycytidine and the pharmacologically acceptable salts thereof
US5084445A (en) * 1986-05-01 1992-01-28 University Of Georgia Research Foundation, Inc. 3'-azido-2',3'-dideoxy-5-methylcytidine
US4987224A (en) * 1988-08-02 1991-01-22 University Of Georgia Research Foundation, Inc. Method of preparation of 2',3'-dideoxynucleosides

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US64631A (en) * 1867-05-14 Thomas g
JPS63275597A (ja) * 1987-05-07 1988-11-14 Ajinomoto Co Inc ジデオキシシチジンの製造方法、その製造中間体、およびその製造方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 111, no. 1, 3 July 1989, Columbus, Ohio, US; abstract no. 7759y, M.UCHIYAMA ET AL. 'Preparation of 2',3'-Dideoxycytidine.' page 748 ;column 2 ; & JP-A-63 275 597 (AJINOMOTO CO., INC....) 14 November 1988 *
CHEMICAL ABSTRACTS, vol. 111, no. 11, 11 September 1989, Columbus, Ohio, US; abstract no. 97691x, S.BRODER ET AL. 'Preparation of Antiretroviral 2',3'-Dideoxynucleoside Dimers Effective Against Human Immunodeficiency Virus (HIV).' page 792 ;column 2 ; & US-A-64 631 (UNITED STATES DEPT. OF HEALTH AND HUMAN SERVICES.) 1 September 1988 *
JOURNAL OF ORGANIC CHEMISTRY., vol.53, 1988, EASTON US pages 4780 - 4786 M.OKABE ET AL. 'Synthesis of the Dideoxynucleosides ddC and CNT from Glutamic Acid, Ribonolactone, and Pyrimidine Bases.' *
See also references of WO9323413A1 *

Also Published As

Publication number Publication date
CA2135498A1 (fr) 1993-11-25
MX9302761A (es) 1994-05-31
AU4234193A (en) 1993-12-13
EP0640091A4 (fr) 1995-07-26
WO1993023413A1 (fr) 1993-11-25
JPH07508022A (ja) 1995-09-07

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