Title:
SELECTIVE SOLVENT EXTRACTION FOR THE PURIFICATION OF PROTECTED NUCLEOSIDES
Document Type and Number:
WIPO Patent Application WO/2000/039138
Kind Code:
A1
Abstract:
A process for the purification of nucleosides, and more particularly to a selective solvent extraction method for purifying protected nucleosides. In the purification process, solid particles of protected nucleosides are selectively washed to remove undesirable polar and/or non-polar impurities, while leaving the solid particles substantially undissolved.
Inventors:
SWEDO RAYMOND J
KOLJACK MATHIAS P
GAITA ROMULUS
YATES STEPHEN F
WU CHARLES CHAO
KOLJACK MATHIAS P
GAITA ROMULUS
YATES STEPHEN F
WU CHARLES CHAO
Application Number:
PCT/US1999/030412
Publication Date:
July 06, 2000
Filing Date:
December 21, 1999
Export Citation:
Assignee:
ALLIED SIGNAL INC (US)
International Classes:
C07H19/06; B01D11/02; C07H19/16; (IPC1-7): C07H1/08
Foreign References:
| US5639867A | 1997-06-17 |
Other References:
G.S. LI, B.L. GAFFNEY, R.A. JONES: "Transient protection: efficient one-flask syntheses of protected deoxynucleosides", J. AM. CHEM. SOC., vol. 104, 1982, pages 1316 - 1319, XP002136167
J.P. HORWITZ, J.A. URBANSKI, J. CHUA: "Nucleosides. II. 5'-O-Mesylthymidine and 3'-O-mesylthymidine", J. ORG. CHEM., vol. 27, 1962, pages 3300 - 3302, XP002136168
H. SCHALLER, G. WEIMANN, B. LERCH, H. KHORANA: "Studies on polynucleotides. XXIV. The stepwise synthesis of specific deoxyribopolynucleotides (4). Protected derivatives of deoxyribonucleosides and new syntheses of deoxyribonucleoside-3' phosphates", J. AM. CHEM. SOC., vol. 85, 1963, pages 3821 - 3827, XP002136169
J.P. HORWITZ, J.A. URBANSKI, J. CHUA: "Nucleosides. II. 5'-O-Mesylthymidine and 3'-O-mesylthymidine", J. ORG. CHEM., vol. 27, 1962, pages 3300 - 3302, XP002136168
H. SCHALLER, G. WEIMANN, B. LERCH, H. KHORANA: "Studies on polynucleotides. XXIV. The stepwise synthesis of specific deoxyribopolynucleotides (4). Protected derivatives of deoxyribonucleosides and new syntheses of deoxyribonucleoside-3' phosphates", J. AM. CHEM. SOC., vol. 85, 1963, pages 3821 - 3827, XP002136169
Attorney, Agent or Firm:
Criss, Roger H. (Attn: A. Olinger 101 Columbia Road P.O. Box
2245 Morristown, NJ, US)
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Claims:
CLAIMS We claim:
| 1. | A method for purifying a composition comprising solid particles of a protected nucleoside and polar and nonpolar impurities, the method comprising: a) applying a solvent in which the polar impurities are soluble and in which the particles of protected nucleoside are at most slightly soluble; and b) applying a solvent in which the nonpolar impurities are soluble and in which the particles of protected nucleoside are at most slightly soluble, wherein steps (a) and (b) may be performed in any order. |
| 2. | The method of claim 1 wherein said solvent for the polar impurities comprises a mixture of miscible first and second polar solvents, wherein the particles of protected nucleoside are soluble in the first polar solvent, and relatively insoluble in the second polar solvent. |
| 3. | The method of claim 2 wherein said first polar solvent is selected from the group consisting of acetonitrile, C, to C6 linear and branched alcohols, dimethylformamide, dimethyl sulfoxide, acetone and mixtures thereof, and said second polar solvent is water. |
| 4. | The method of claim 3 wherein said first polar solvent is acetonitrile. |
| 5. | The method of claim 4 wherein the ratio of acetonitrile to water is within the range of about 5: 95 to about 30: 70 by volume. |
| 6. | The method of claim 1 wherein said solvent for nonpolar impurities comprises a mixture of miscible first and second nonpolar solvents, wherein the particles of protected nucleoside are highly soluble in the first nonpolar solvent, and relatively insoluble in the second nonpolar solvent. |
| 7. | The method of claim 6 wherein said first nonpolar solvent is selected from the group consisting of chlorinated solvents, ethyl acetate and mixtures thereof, and said second nonpolar solvent is selected from the group consisting of C5 to CIO linear, branched and cyclic hydrocarbons and mixtures thereof. |
| 8. | The method of claim 7 wherein said chlorinated solvents are selected from the group consisting of methylene chloride chloroform, carbon tetrachloride, dichloroethane and mixtures thereof. |
| 9. | The method of claim 7 wherein said first nonpolar solvent is methylene chloride. |
| 10. | The method of claim 7 wherein said second nonpolar solvent is hexane or cyclohexane. |
| 11. | The method of claim 7 wherein said first nonpolar solvent is methylene chloride and said second nonpolar solvent is hexane. |
| 12. | The method of claim 11 wherein the ratio of methylene chloride to hexane is within the range of about 3: 1 to about 1: 3, by volume. |
| 13. | The method of claim 12 wherein the ratio of methylene chloride to hexane is about 1: 2, by volume. |
| 14. | The method of claim 1 wherein the nucleoside which is protected is selected from the group consisting of deoxyadenosine, deoxycytidine, deoxyguanosine and thymidine. |
| 15. | The method of claim 1 wherein the nucleoside includes amino and alcohol functional groups, and protection involves derivatization of the amino and alcohol groups. |
| 16. | The method of claim 15 wherein the alcohol group is derivatized as a trityl or dimethoxytrityl ether. |
| 17. | The method of claim 15 wherein the amino group is derivatized as a benzoyl or isobutyryl group. |
| 18. | The method of claim 15 wherein the protected nucleoside is selected from the group consisting of N6benzoyl5'O (4,4'dimethoxytrityl)2'deoxyadenosine and N2 isobutyryl5'0 (4,4'dimethoxytrityl)2'deoxyguanosine. |
| 19. | The method of claim 1 wherein step (a) is performed before step (b). |
| 20. | The method of claim 1 further comprising recovering purified particles of protected nucleoside. |
| 21. | The method of claim 20 wherein said recovering comprises filtering from the purified particles from the solvent of the last applying step. |
| 22. | The method of claim 21 further comprising evaporating the solvent from the filtered particles. |
| 23. | A method for purifying a composition comprising solid particles of a protected nucleoside and impurities, the method comprising applying a solvent in which the impurities are soluble and in which the particles of protected nucleoside are at most slightly soluble. |
| 24. | The method of claim 23 in which the impurities comprise polar and nonpolar impurities. |
| 25. | The method of claim 24 wherein said solvent comprises a mixture of a polar solvent and a nonpolar solvent. |
| 26. | The method of claim 25 wherein said solvent further comprises a solvent which is miscible with both the polar and nonpolar solvents. |
| 27. | The method of claim 26 wherein said solvent comprises hexane, methanol and methylene chloride. |
| 28. | The method of claim 23 wherein the nucleoside which is protected is selected from the group consisting of deoxyadenosine, deoxycytidine, deoxyguanosine and thymidine. |
| 29. | The method of claim 28 wherein the solvent comprises water. |
| 30. | The method of claim 28 wherein the solvent comprises water and acetonitrile. |
| 31. | The method of claim 28 wherein the solvent comprises methanol and hexane. |
| 32. | The method of claim 28 wherein the solvent comprises hexane, methanol and methylene chloride. |
Description:
INTERNATIONALSEARCHREPORT IntemationalApplicationNo PCT/US99/30412 C.(Continuation)DOCUMENTSCONSIDEREDTOBERELEVANT Category'Citation ofdocument,withindication,whereappropriate,oftherelevantpass
agesRelevanttoclaimNo. XUS5639867A(BRILLWOLFGANGK-D)23,24,28 17June1997(1997-06-17) column13,line10-line13 column16,line48-line53 column17,line12-line18 XH.SCHALLER,G.WEIMANN,B.LERCH,H.23,24,28 KHORANA:"Studiesonpolynucleotides. XXIV.Thestepwisesynthesisofspecific deoxyribopolynucleotides(4).Protected derivativesofdeoxyribonucleosidesand newsynthesesofdeoxyribonucleoside-3' phosphates" J.AM.CHEM.SOC., vol.85,1963,pages3821-3827, XP002136169 citedintheapplication page3823,left-handcolumn,paragraph6 page3823,right-handcolumn,paragraph1 6 In.iationalapplicationNo. INTERNATIONALSEARCHREPORTPCT/US99/30412 Box)Observationswherecertainclaimswerefoundunsearchable(Cont
inuationofitem1offirstsheet) ThisInternationalSearchReporthasnotbeenestablishedinrespecto
fcertainclaimsunderArticle17(2)(a)forthefollowingreasons: 1.ClaimsNos.: becausetheyrelatetosubjectmatternotrequiredtobesearchedbythi
sAuthority,namely: 2.YClaimsNos.:23(partially) becausetheyrelatetopartsoftheInternationalApplicationthatdon
otcomplywiththeprescribedrequirementstosuch anextentthatnomeaningfulInternationalSearchcanbecarriedout,s
pecifically: seeFURTHERINFORMATIONsheetPCT/ISA/210 3.ClaimsNos.: becausetheyaredependentclaimsandarenotdraftedinaccordancewit
hthesecondandthirdsentencesofRule6.4(a). BoxIfObservationswhereunityofinventionislacking(Continuation
ofitem2offirstsheet) ThisIntemationalSearchingAuthorityfoundmultipleinventionsint
hisinternationalapplication,asfollows: 1.As ai !requiredadditionalsearchfeesweretimelypaidbytheapplicant,th
isinternationalSearchReportcoversall searchableclaims. 2.As all searchable claims couldbesearchedwithouteffortjustifyinganadditionalfee,thisAu
thoritydidnotinvitepayment ofanyadditionalfee. 3.Asonlysomeoftherequiredadditionalsearchfeesweretimelypaidb
ytheapplicant,thisInternationalSearchReport covers onlythoseclaimsforwhichfeeswerepaid,specificallyclaimsNos.: 4.Norequiredadditionalsearchfeesweretimelypaidbytheapplicant
.Consequently,thisInternationalSearchReportis restrictedtotheinventionfirstmentionedintheclaims ;itiscoveredbyclaimsNos.: RemarkonProtestThe additionalsearchfeeswereaccompaniedbytheapplicant'sprotest. u 7j Noprotestaccompaniedthepaymentofadditionalsearchfees. u FURTHER INFORMATION CONTINUED FROM PCT/ISA/210 Continuation of Box 1.2 Claims Nos.: 23 (partially) Present claim 23 relate to an extremely large number of possible methods.
Support within the meaning of Article 6 PCT and/or disclosure within the meaning of Article 5 PCT is to be found, however, for only a very small proportion of the methods claimed. In the present case, claim 23 so lack support, and the application so lacks disclosure, that a meaningful search over the whole of the claimed scope is impossible. Consequently, the search of claim 23 has been carried out in the light of those parts relating to the methods disclosed in the examples.
The applicant's attention is drawn to the fact that claims, or parts of claims, relating to inventions in respect of which no international search report has been established need not be the subject of an international preliminary examination (Rule 66.1 (e) PCT). The applicant is advised that the EPO policy when acting as an International Preliminary Examining Authority is normally not to carry out a preliminary examination on matter which has not been searched. This is the case irrespective of whether or not the claims are amended following receipt of the search report or during any Chapter II procedure. INTERNATIONALSEARCHREPORT------------------- International ApplicationNo ) rationonpatentfamilymembers PC, PC./US99/30412 PatentdocumentPublicationPatentfamily Publication citedinsearchreportdatemember(s)date US5639867A17-06-1997AT159949T15-11-1997 AU671387B22-08-1996 AU7304394A30-03-1995 CA2132133A18-03-1995 CN1111249A08-11-1995 DE59404522D11-12-1997 EP0644196A22-03-1995 ES2110726T16-02-1998 JP7165787A27-06-1995 ZA9407180A17-03-1995