Methods of creating and screening DNA-encoded libraries
The present invention features a number of methods for identifying one or more compounds that bind to a biological target. The methods include synthesizing a library of compounds, wherein the compounds contain a functional moiety having one or more diversity positions. The functional moiety of the compounds is operatively linked to an initiator oligonucleotide that identifies the structure of the functional moiety.
1. A method of tagging DNA-encoded chemical libraries, said method comprising:
(a) binding a first functional group of a bifunctional linker to a component of said chemical library, wherein said component comprises a functional moiety,
(b) binding a second functional group of a bifunctional linker to an initiator oligonucleotide at the 5′ end of said initiator oligonucleotide, and
(c) binding an identifier region at the 3′ end of said initiator oligonucleotide,
wherein said initiator oligonucleotide is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions,
wherein said modified initiator oligonucleotide comprises a nucleotide having a hydrophobic moiety, and said hydrophobic moiety is an aliphatic chain at the C5 position.
2. A method of tagging DNA-encoded chemical libraries, said method comprising:
(a) binding a first functional group of a bifunctional linker to a component of said chemical library, wherein said component comprises a functional moiety,
(b) binding a second functional group of a bifunctional linker to an initiator oligonucleotide at the 5′ end of said initiator oligonucleotide, and
(c) binding an identifier region at the 3′ end of said initiator oligonucleotide,
wherein said initiator oligonucleotide is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions,
wherein said modified initiator oligonucleotide comprises an insertion having a hydrophobic moiety, and said hydrophobic moiety is an azobenzene.
3. The method of claim 1 , wherein said initiator oligonucleotide bound to said bifunctional linker forms a hairpin structure.
4. The method of claim 2 , wherein said initiator oligonucleotide bound to said bifunctional linker forms a hairpin structure.
5. The method of claim 1 , wherein said initiator oligonucleotide comprises an identifier region.
6. The method of claim 5 , wherein said identifier region of step (c) hybridizes to said identifier region of said initiator oligonucleotide.
7. The method of claim 2 , wherein said initiator oligonucleotide comprises an identifier region.
8. The method of claim 7 , wherein said identifier region of step (c) hybridizes to said identifier region of said initiator oligonucleotide.
9. The method of claim 1 , wherein said bifunctional linker is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions.
10. The method of claim 9 , wherein the modified bifunctional linker comprises one or more of an alkyl chain, a polyethylene glycol unit, a branched species with positive charges, or a hydrophobic ring structure.
11. The method of claim 10 , wherein the modified bifunctional linker comprises 12 to 45 polyethylene glycol units.
12. The method of claim 2 , wherein said bifunctional linker is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions.
13. The method of claim 12 , wherein the modified bifunctional linker comprises one or more of an alkyl chain, a polyethylene glycol unit, a branched species with positive charges, or a hydrophobic ring structure.
14. The method of claim 13 , wherein the modified bifunctional linker comprises 12 to 45 polyethylene glycol units.
15. The method of claim 1 , wherein said identifier region of step (c) or identifier region of said initiator oligonucleotide is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions.
16. The method of claim 2 , wherein said identifier region of step (c) or identifier region of said initiator oligonucleotide is modified to increase solubility of a member of said DNA-encoded chemical library in organic conditions.
17. The method of claim 1 , wherein said identifier region of step (c) encodes said functional moiety.
18. The method of claim 2 , wherein said identifier region of step (c) encodes said functional moiety.
19. The method of claim 1 , wherein prior to step (c) the method comprises linking a chemical building block to said functional moiety and said identifier region of step (c) encodes said chemical building block.
20. The method of claim 2 , wherein prior to step (c) the method comprises linking a chemical building block to said functional moiety and said identifier region of step (c) encodes said chemical building block.
21. The method of claim 1 , wherein said member of said DNA-encoded chemical library has an octanol:water coefficient from 1.0 to 2.5.
22. The method of claim 2 , wherein said member of said DNA-encoded chemical library has an octanol:water coefficient from 1.0 to 2.5.