Encoded combinatorial chemical libraries

• US 6,060,596 A
• Filed: 03/03/1998
• Issued: 05/09/2000
• Est. Priority Date: 03/30/1992
• Status: Expired due to Term

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Brenner, Sydney, Lerner, Richard, Janda, Kim
• Primary Examiner(s): YUCEL, IREM
• Application Number:
  US09/033,743
• Time in Patent Office: 798 Days
• Field of Search: 536/25.3, 536/24.2, 436/518, 436/536, 435/6
• US Class Current: 506/15
• CPC Class Codes: B01J 2219/00572 Chemical means
• 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
• CPC Class Codes: C12N 15/1062 mRNA-Display, e.g. polypeptide and encoding template are connected covalently
• CPC Class Codes: C12N 15/1075 by coupling phenotype to genotype, not provided for in other groups of this subclass
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes

Abstract

The present invention describes an encoded combinatorial chemical library comprised of a plurality of bifunctional molecules having both a chemical polymer and an identifier oligonucleotide sequence that defines the structure of the chemical polymer. Also described are the bifunctional molecules of the library, and methods of using the library to identify chemical structures within the library that bind to biologically active molecules in preselected binding interactions.

Claims

1. A bifunctional molecule according to the formula A--B--C, wherein A is a polymer comprising a linear series of chemical units represented by the formula (X_n)_a, wherein X is a single chemical unit in polymer A, B is a linker molecule operatively linked to A and C. and identifier oligonucleotide C is represented by the formula (Z_n)_a, wherein a unit identifier nucleotide sequence Z within oligonucleotide C identifies the chemical unit X at position n;

and wherein n is a position identifier for both X in polymer A and Z in oligonucleotide C having the value of 1+i where i is an integer from 0 to 10, such that when n is 1, X or Z is located most proximal to the linker, and a is an integer from 4 to 50.

2. The bifunctional molecule of claim 1 wherein said unit identifier nucleotide sequence Z has a length of from 2 to 8 nucleotides.

3. The bifunctional molecule of claim 1 wherein said polymer is an oligosaccharide, polepeptide, glycolipid, lipid, proteoglycan, glycopeptide or oligonucleotide.

4. The bifunctional molecule of claim 1 wherein said polymer A is a polypeptide, X is an amino acid residue in said polypeptide, and unit identifier nucleotide sequence Z is a hexanucleotide sequence that identifies the amino acid residue at position n in polypeptide A.

5. The bifunctional molecule of claim 4 wherein said amino acid residue is selected from the group consisting of natural, modified and non-natural amino acids.

6. The bifunctional molecule of claim 1 wherein said identifier oligonucleotide C has a nucleotide sequence according to the formula P1--(Z_n)_a --P2, where P1 and P2 are nucleotide sequences that provide polymerase chain reaction (PCR) primer binding sites adapted to amplify the identifier oligonucleotide.

7. The bifunctional molecule of claim 6 wherein said P1 and P2 each contain a sequence that defines a restriction endonuclease site when present in a PCR-amplified duplex DNA fragment.

8. The bifunctional molecule of claim 7 wherein said restriction endonuclease sites are located proximal to (Z_n)_a relative to the PCR primer binding sites.

9. The bifunctional molecule of claim 8 wherein said restriction endonuclease sites form non-overlapping cohesive termini upon restriction endonuclease cleavage.

10. A library comprising a plurality of species of bifunctional molecules according to claim 1.

11. The library of claim 10 wherein said plurality of species is defined by the formula V^a, where V represents the number of different chemical units forming an alphabet of possible chemical units of X, and a is an exponent to V and represents the number of chemical units of X forming polymer A.

12. The library of claim 11 wherein X is a natural amino acid and V is 20.

13. The library of claim 11 wherein the number of chemical units (a) forming polymer A is from about 3 to about 8.

14. The library of claim 12 wherein a is 6.

15. The library of claim 12 wherein X is an amino acid and identifier oligonucleotide C is represented by the formula (Z_n)_a wherein a unit identifier nucleotide sequence Z within oligonucleotide C identifies the chemical unit X at position n:

and wherein n is a position identifier for both X in polymer A and Z in oligonucleotide C having the value of 1+i where i is an integer from 0 to 10, such that when n is 1, X or Z is located most proximal to the linker, and a is an integer from 4 to 50 and said unit identifier nucleotide sequence Z has a length from 3 to 6 nucleotides.

16. The library of claim 10 wherein each of said species of bifunctional molecules in said plurality is present in molar equivalents of from 0.2 to 10.0.

17. The library of claim 11 wherein said identifier oligonucleotide in each of said species of bifunctional molecules has a nucleotide sequence according to the formula P1--(Z_n)_a --P2, where P1 and P2 are nucleotide sequences that provide PCR primer binding sites adapted to amplify the identifier oligonucleotide, and where the nucleotide sequences of P1 and P2 are shared by all bifunctional molecule species in the library.