Modified nucleotide compounds

• US 7,495,088 B1
• Filed: 06/28/1994
• Issued: 02/24/2009
• Est. Priority Date: 12/04/1989
• Status: Active Grant

• Current Assignee: Enzo Biochem Incorporated
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Quartin, Robin S., Brakel, Christine L., Wetmur, James G.
• Primary Examiner(s): Zhou; Shubo (Joe)
• Application Number:
  US08/479,999
• Time in Patent Office: 5,355 Days
• Field of Search: 536/27, 536/22.1, 536/23.1, 536/24.1, 536/25.3, 536 243- 2433, 435/6, 435/810, 436/501, 935/77, 935/78
• US Class Current: 536/23.1
• CPC Class Codes: A61P 31/12 Antivirals
• 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

Disclosed is a nuclease resistant nucleotide compound capable of hybridizing with a complementary RNA in a manner which inhibits the function thereof, which modified nucleotide compound includes at least one component selected from the group consisting of MN₃M, B(N)_xM and M(N)_xB wherein N is a phosphodiester-linked modified 2′-deoxynucleoside moiety; M is a moiety that confers endonuclease resistance on said component and that contains at least one modified or unmodified nucleic acid base; B is a moiety that confers exonuclease resistance to the terminus to which it is attached; and x is an integer of at least 2.

Claims

1. A modified nucleotide compound which contains at least one sequence having the formula MN₃M wherein N is a phosphodiester-linked unmodified 2′

-deoxynucleoside moiety containing at least one guanine, adenine, cytosine or thymine moiety and M is a methylphosphonate-containing deoxynucleotide.

2. A method of inhibiting the function of an RNA, which comprises:

contacting said RNA, under conditions permissive of hybridization, with a modified nucleotide compound which includes at least one sequence having the formula MN₃M wherein N is a phosphodiester-linked unmodified 2′

-deoxynucleoside moiety containing at least one guanine, adenine, cytosine or thymine moiety and M is a methylphosphonate-containing deoxynucleoside.

3. A method of identifying a nucleotide compound having a combination of nuclease resistance and the ability to form an RNase H substrate when in complex with an RNA, which method comprises:

(i) preparing modified nucleotide compounds;

(ii) selecting by exo-and endonuclease digestion those modified nucleotide compounds of (i) which are nuclease-resistant as shown by being capable of forming and electrophoretically migrating as a duplex with a complementary nucleotide compound; and

(iii) selecting by RNase H digestion those of the nuclease-resistance nucleotide compounds of (ii) which act as substrates for RNase H when hybridized with a complementary RNA.