Details for Patent: 5,731,424

Title:	High affinity TGF.beta. nucleic acid ligands and inhibitors
Abstract:	Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGF.beta.. Included in the invention are specific RNA and ssDNA ligands to TGF.beta.1 identified by the SELEX method. Also included are RNA ligands that inhibit the interaction of TGF.beta.1 with its receptor.
Inventor(s):	Toothman; Penelope J. (Boulder, CO), Ringquist; Steven (Lyons, CO), Gold; Larry (Boulder, CO)
Assignee:	NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:	Jun 02, 1995
Application Number:	08/458,424
Claims:	1. A nucleic acid ligand to transforming growth factor-beta (TGF.beta.) identified according to a method comprising: a) preparing a candidate mixture of nucleic acids; b) contacting the candidate mixture of nucleic acids with TGF.beta., wherein nucleic acids having an increased affinity to TGF.beta. relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to TGF.beta., whereby said nucleic acid ligand of TGF.beta. may be identified. 2. A nucleic acid ligand to transforming growth factor-beta 1 (TFG.beta.1) identified according to a method comprising: a) preparing a candidate mixture of nucleic acids; b) contacting the candidate mixture of nucleic acids with TGF.beta.1, wherein nucleic acids having an increased affinity to TGF.beta.1 relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to TGF.beta.1, whereby said nucleic acid ligand of TGF.beta.1 may be identified. 3. A purified and isolated non-naturally occurring RNA ligand to TGF.beta.1 wherein said ligand is selected from the group consisting of the sequences set forth in Table 3 (SEQ ID NOS: 12-42). 4. The nucleic acid ligand of claim 1 wherein in the method, said candidate mixture of nucleic acids is comprised of single stranded nucleic acids. 5. The nucleic acid ligand of claim 4 wherein in the method, said single stranded nucleic acids are ribonucleic acids. 6. The nucleic acid ligand of claim 4 wherein in the method, said single stranded nucleic acids are deoxyribonucleic acids. 7. A purified and isolated non-naturally occurring ssDNA ligand to TGF.beta.1 wherein said ligand is selected from the group consisting of the sequences set forth in Table 6 (SEQ ID NOS: 55-89). 8. A purified and isolated non-naturally occurring nucleic acid ligand to transforming growth factor-beta (TGF.beta.). 9. A purified and isolated non-naturally occurring nucleic acid ligand to transforming growth factor-beta 1 (TGF.beta.1). 10. The nucleic acid ligand of claim 5 wherein in the method, said single stranded nucleic acids are comprised 2'-amino (2'-NH.sub.2) modified ribonucleic acids. 11. The nucleic acid ligand of claim 5 wherein in the method, said single stranded nucleic acids are comprised 2'-fluoro (2'-F) modified ribonucleic acids.