Method of preparing an adduct

Abstract

A method for identifying one or several molecular structure(s) having a high-affinity for a target of interest, the molecular structure(s) each including one nucleotide chain onto which is hybridized at least one PNA-encoded molecule.

Description

[0001]The present invention relates to methods for identifying one or several consensus structure(s) having a high-affinity for a chemical target of interest, and to a method of preparing an adduct starting from said potential consensus structure(s).[0002]The discovery of selective ligands to a protein remains a rate limiting step in drug discovery. While classical high throughput screening has proven successful for the screening of libraries of tens of thousands to hundreds of thousands of compounds, it is prohibitively expensive and cumbersome to scale out this technology on a proteomic scale. The approach proposed herein aims to overcome these limitations in providing a technology which enables the selection and evolution of small molecules from readily available libraries containing up to millions of compounds obtained by self assembly of PNA-encoded libraries onto DNA-templates in a highly miniaturized format. The inventors have developed technologies to synthesize PNA-encoded ...

AI Technical Summary

Benefits of technology

[0007]There is consequently a need for a more rapid, easy and economic technology for the selection of fittest ligands for a specific target.

[0008]The inventors have discovered a way to capitalize on the programmable assembly of these PNA-tagged small molecules onto libraries of DNA templates to be able to amplify the nucleic acid tag by PCR. This technique allows a very sensitive detection of the selected entity by virtue of the exponential amplification of the PCR reaction. An additional advantage of displaying PNA-encoded small molecules onto DNA templates is that it allows to combinatorially display small molecules as fragments of a ligand.

[0023]By the expression “PNA(s)-nucleotide chain hybrid” is meant a compound which consists of a nucleotide chain onto which at least one, but preferably two PNA-encoded molecule(s) or multiple PNA-encoded molecules are hybridized. According to the invention, the resulting library of PNA(s)-nucleotide chain hybrids self assembles into predictable hybrids based on the rules of hybridization and nucleotide recognition. The identity of the molecules(s) can be inferred by the sequence of the nucleotide chain. Hybridization of multiple PNA-encoded molecules onto a nucleotide chain enables a combinatorial display of the PNA-encoded molecules which can interact cooperatively with a target of interest.

Problems solved by technology

The discovery of selective ligands to a protein remains a rate limiting step in drug discovery.

While classical high throughput screening has proven successful for the screening of libraries of tens of thousands to hundreds of thousands of compounds, it is prohibitively expensive and cumbersome to scale out this technology on a proteomic scale.

In all cases, the diversification and encoding reactions must be performed in water under conditions which do not degrade the DNA which in itself is a severe limitation on the repertoire of reactions that can be utilized to construct the libraries.

Further limitations for technologies which rely on an enzymatic transformation to extend the nucleic acid tag (Praecis and Phylochem) is the possibility of enzyme poisoning with small molecules present in the library or reagents left from the previous transformations.

While this approach does provide N-terminus appended libraries, it does not allow these libraries to be prepared in a split and mix fashion.

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