Regulated aptamer therapeutics

• US 7,960,102 B2
• Filed: 11/30/2005
• Issued: 06/14/2011
• Est. Priority Date: 07/25/2002
• Status: Active Grant

• Current Assignee: Archemix Corp. (Baxter International Inc.)
• Original Assignee: Archemix Corp. (Baxter International Inc.)
• Inventors: Epstein, David, Wilson, Charles, Diener, John L.
• Primary Examiner(s): Chong; Kimberly
• Application Number:
  US11/291,610
• Publication Number:
  US 20060084109A1
• Time in Patent Office: 2,022 Days
• Field of Search: None
• US Class Current: 435/6.11
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/1048 SELEX
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/16 Aptamers
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2320/50 Methods for regulating/modulating their activity
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C12Q 2541/101 Selex
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose

Abstract

Materials and methods of use thereof are presented for the treatment of diabetes and other diseases. Therapeutic compositions including regulated aptamer therapeutic compositions with specificity to components of diabetes disease are presented with methods of administering these therapeutic compositions.

Claims

1. A method for selecting a regulated aptamer comprising the steps of:

a) identifying a first pool of candidate aptamers comprising a first binding domain that binds to a first ligand, wherein the first ligand is selected from the group consisting of a small molecule, a peptide, a protein and a saccharide, and a second pool of candidate aptamers comprising a second binding domain that binds to a second ligand, wherein the second ligand is a protein or a peptide;

b) engineering a chimeric pool of aptamers wherein individual aptamers within the chimeric pool comprise a first binding domain of a candidate aptamer from the first candidate pool linked to a second binding domain of a candidate aptamer from the second candidate pool; and

c) selecting, using SELEX, an aptamer from the chimeric pool wherein binding of the second ligand by the second binding domain of the selected aptamer is activated by binding of the first ligand by the first binding domain, and wherein binding of the second ligand by the second binding domain modulates a biological activity of the second ligand.

2. The method of claim 1, wherein the identifying step further comprises using SELEX to isolate the first pool of candidate aptamers.

3. The method of claim 1, wherein the identifying step further comprises using SELEX to isolate the second pool of candidate aptamers.

4. The method of claim 1, wherein the selecting step further comprises contacting the chimeric pool of aptamers with the second ligand, in a negative selection step, to result in a bound ligand and unbound ligand mixture, and partitioning the unbound ligand mixture;

contacting only the unbound ligand mixture, in a positive selection step, with the first and second ligands to result in a second bound ligand mixture and unbound ligand mixture; and

selecting an aptamer from the second bound ligand mixture.

5. The method of claim 4, wherein the selecting step further comprises amplifying and contacting the second bound ligand mixture with the second ligand in the negative selection step and repeating the negative and positive selection steps iteratively prior to selecting the aptamer.

6. The method of claim 1, wherein the engineering step comprises linking the first and second binding domains of the first and second candidate aptamers by linearly concatenating the first and second ligand binding domains with an intervening random sequence domain.

7. The method of claim 1, wherein the engineering step comprises linking the first and second binding domains of the first and second candidate aptamers by coupling the first and second binding domains with linking helices.

8. The method of claim 1, wherein the first ligand is a protein selected from the group consisting of a growth factor, a transcription factor, an enzyme, an immunoglobulin, a receptor and a hormone.