Claims
- 1. A method of deprotecting an oligonucleotide, which method comprises heating an oligonucleotide of the formula:
- 2. The method of claim 1, wherein Q or Q1 comprises a nucleoside of the formula:
- 3. The method of claim 1, wherein R1 is H, an alkyl or NR1aR1a′, wherein R1a and R1a′, together with the nitrogen atom to which they are bonded, comprise a heterocycle containing from 3 to about 7 atoms in the ring skeleton thereof.
- 4. The method of claim 1, wherein X1 is S.
- 5. The method of claim 1, wherein Z is CR2aR2a′ or CR2aR2a′CR2bR2b′, and R2a, R2a′, R2b and R2b′ are the same or different and each is H or an alkyl.
- 6. The method of claim 1, wherein R2 or R2′ is H or an alkyl.
- 7. The method of claim 1, wherein R3 or R3′ is H, an alkyl or an aryl.
- 8. The method of claim 1, wherein R is a protecting group of the formula:
- 9. The method of claim 1, wherein the temperature is from about 50° C. to about 90° C.
- 10. The method of claim 1, wherein the deprotection is carried out at about pH 7.
- 11. The method of claim 1, wherein the fluid medium contains water.
- 12. A compound selected from the group consisting of compounds of the formulae:
- 13. The compound of claim 12, wherein Q or Q1 comprises a nucleoside of the formula:
- 14. The compound of claim 12, wherein R1 is H, an alkyl or NR1aR1a′, wherein R1a and R1a′, together with the nitrogen atom to which they are bonded, comprise a heterocycle containing from 3 to about 7 atoms in the ring skeleton thereof.
- 15. The compound of claim 12, wherein X1 is S.
- 16. The compound of claim 12, wherein Z is CR2aR2a′ or CR2aR2a′CR2bR2b′ and R2a, R2a′, R2b and R2b′ are the same or different and each is H or an alkyl.
- 17. The compound of claim 12, wherein R2 or R2′ is H or an alkyl.
- 18. The compound of claim 12, wherein R3 or R3′ is H, an alkyl or an aryl.
- 19. The compound of claim 12, wherein R is a protecting group of the formula:
- 20. A method of producing an oligonucleotide, which method comprises
(a) reacting a nucleophile of the formula: R4—O—Q1—OHwith an electrophile of the formula: 51wherein W is a dialkylamino group that is displaced by the nucleophile, under conditions to displace W and produce an adduct comprising a tricoordinated phosphorus atom; (b) reacting the product obtained in step (a) with a reagent selected from the group consisting of oxidizing agents, sulfurizing agents, and selenizing agents to produce a protected oligonucleotide of the formula: 52wherein n=1; (c) cleaving R15 from the protected oligonucleotide from step (b) to produce a nucleophile; (d) optionally repeating steps (a)-(c) until an oligomer of a specified length is obtained; and (e) heating the product from step (c) or (d) in a fluid medium, at a substantially neutral pH, at a temperature up to about 100° C. to produce a deprotected oligonucleotide of the formula: 53wherein R is a thermolabile protecting group of the formula: 54R1 is H, R1a, OR1a, SR1a or NR1aR1a′, wherein R1a and R1a′ are the same or different and each is H, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, or an aralkyl; or, when R1 is NR1aR1a′, R1a and R1a′, together with the nitrogen atom to which they are bonded, comprise a heterocycle containing from 3 to about 7 atoms in the ring skeleton thereof, X is O, S or Se; X is O or S; Z is O, S, NR2a, CR2aR2a′ or CR2aR2a′CR2bR2b′, wherein R2a, R2a′, R2b and R2b′ are the same or different and each is H, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, or an aralkyl; or R1a or R1a′, in combination with any of R2a, R2a′, R2b or R2b′, together with C═X of the protecting group to which they are bonded, comprise a ring containing from 3 to about 7 atoms in the skeleton thereof; provided that R1 is not R1a when Z is S, Z is not CR2aR2a′ or CR2aR2a′CR2bR2b′ when R1 is SR1a, and Z is not O or S when R1 is H; R2, R2′, R3 and R3′ are the same or different and each is H, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, or an aralkyl, or R2 or R2′, in combination R3 or R3′, together with the carbon atoms to which they are bonded, comprise a cyclic substituent of the formula: 55wherein p is an integer from 0-6 and a-d are the same or different and each is selected from the group consisting of H, an alkyl, a nitro, an amino, a hydroxyl, a thio, a cyano and a halogen, provided that the aromatic ring, which bears the substituents a-d, is one carbon removed from the phosphate oxygen of formula (IIIa), wherein R1, R2a, R2a′, R2b, R2b′, R2, R2′, R3 or R3′ is unsubstituted or substituted with one or more substituents, which are the same or different, selected from the group consisting of OR8, CN, NO2, N3, and a halogen, wherein R8 is H or an alkyl; R4 and R15 are the same or different and each is H, a hydroxyl protecting group, or a solid support; Q1 is a nucleoside, an oligonucleotide or an oligomer comprising an oligonucleotide; n is an integer from 1 to about 300; and Q is a nucleoside, an oligonucleotide or an oligomer comprising an oligonucleotide and, when n is greater than 1, each Q is independently selected.
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent application is a continuation-in-part of copending international patent application No. PCT/US00/04032, filed Feb. 16, 2000, pending, which claims priority to U.S. provisional patent application No. 60/125,867, filed Mar. 24, 1999.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60125867 |
Mar 1999 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
PCT/US00/04032 |
Feb 2000 |
US |
Child |
09792799 |
Feb 2001 |
US |