Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof

• US 4,469,863 A
• Filed: 11/12/1980
• Issued: 09/04/1984
• Est. Priority Date: 11/12/1980
• Status: Expired due to Term

• Current Assignee: Lillian Bonnie Murakoshi, Patricia Anne Finch, Robin Lee Finch, Ruth Mae Finch, Vida Marilena Mcarty, Walter G. Finch
• Original Assignee: Lillian Bonnie Murakoshi, Patricia Anne Finch, Robin Lee Finch, Ruth Mae Finch, Vida Marilena Mcarty, Walter G. Finch
• Inventors: Ts'o, Paul O. P., Miller, Paul S.
• Primary Examiner(s): Hazel, Blondel
• Application Number:
  US06/206,297
• Time in Patent Office: 1,392 Days
• Field of Search: 536/27, 536/28, 536/29
• US Class Current: 536/24.5
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

Oligonucleoside alkyl-- or aryl phosphonates are nonionic analogues of nucleic acid which possess unique physical and biological properties. These properties enable the analogues to enter living cells intact and to bind with specifically selected nucleic acids within the cell. As a result, the analogues can specifically inhibit the function or expression of a preselected nucleic acid sequence. Thus the analogues could be used to specifically inhibit the growth of tumor cells or replication of viruses in infected cells.

Four methods are provided for preparing oligonucleoside methylphosphonates: (1) Coupling a protected nucleoside 3'"'"'-alkyl or aryl phosphonate with the 5'"'"'-hydroxyl group of a protected nucleoside using a condensing agent; (2) Coupling protected nucleoside 3'"'"'-alkyl or aryl phosphonic acid derivative with the 5'"'"'-hydroxy group of a protected nucleoside with the activated alkyl or aryl phosphonic acid derivative possessing functionalities which are good leaving groups; (3) Coupling a protected nucleoside 3'"'"'-alkyl or aryl phosphinate derivative with the 5'"'"'-hydroxyl group of a protected nucleoside with the resulting phosphinate derivative being then oxidized to the phosphonate; and (4) Converting a oligonucleoside methoxyphosphite derivative to the alkyl or aryl phosphonate derivative by reaction with an alkyl or aryl iodide. It has been demonstrated that procedures (1) and (2) can be used to prepare oligonucleoside methylphosphonates. Others have shown that procedure (4) can be used to prepare a diribonucleoside methylphosphonate.

Claims

1. Alkyl or aryl phosphonate nucleic acid analogs, comprising, at least five nucleosides, and an alkyl or aryl phosphonate group, said nucleosides being linked together by said alkyl or aryl phosphonate group to form phosphonate nucleic acid analogs, where the alkyl phosphonate group or the aryl phosphonate group do not sterically hinder the phosphonate linkage or interact with each other, said alkyl or aryl phosphonate nucleic acid analog as defined by the structural formula:

##STR9## where B is a base;

R'"'"' is a hydrogen, hydroxyl, O-alkyl or O-aryl or O-halogen;

R is alkyl or aryl; and

where the S and R configurations define the spatial location of the alkyl or aryl group, with R denoting the alkyl or aryl group in a pseudoequitorial position and S denoting the alkyl or aryl group in a pseudoaxial position.

2. The alkyl or aryl phosphonate nucleic acid analog as recited in claim 1, wherein each said nucleoside includes a 3'"'"'-hydroxyl group and a 5'"'"'-hydroxyl group, with a 3'"'"'-hydroxyl group of one nucleoside being linked with a 5'"'"'-hydroxyl group of the other nucleoside.

3. The alkyl or aryl phosphonate nucleic acid analog as recited in claim 1, wherein each said nucleoside has one of the following sugars:

ribose, 2'"'"'-deoxyribose, 2'"'"'-O alkyl ribose, 2'"'"'halogenoribose, or aryl ribose.

4. The alkyl or aryl phosphonate nucleic acid analog as recited in claim 1, wherein each said nucleoside has a base consisting of at least one of the following:

adenine, thymine, cytosine, guanine, uracil or hypoxanthine.

5. The alkyl or aryl phosphonate nucleic acid analog as recited in claim 1, wherein the number of nucleosides can range from at least five to at least twenty linked together by said alkyl or aryl phosphonate groups.

6. The alkyl or aryl phosphonate nucleic acid analog as recited in claim 1, wherein said alkyl or aryl phosphonate group consists of R and S configurations, wherein R and S define the spatial location of the alkyl or aryl group, with R denoting the alkyl or aryl group in a pseudoequitorial position and S denoting the alkyl or aryl group in a pseudoaxial position.

7. A heptadeoxyribonucleoside methylphosphonate nucleic acid analog comprising six methylphosphonate groups, and seven nucleosides, covalently linked together in the following order:

deoxyadenosine, deoxyquanosine, deoxyguanosine, deoxyadenosine, deoxyguanosine, deoxyguanosine, and thymidine, as defined by the structural formula;

##STR10## where Ad is adenine, Gu is guanine, and Th is thymine.