Triazines and pyrimidines as protein binding ligands

Inventors

Betley, Jason Richard • Pearson, James Christopher • Tatton, Helen Rosemary • Beacom, Ben Martin

Assignees

Astrea UK Services Ltd

Abstract

Compounds of the general formula (I): in which inter alia Q1 represents —NR1R3, —OR1 or —SR1 and Q2 represents —NR2R4, —OR2 or —SR2, and A represents the point of attachment to a support matrix, are useful as protein binding ligands when (a) at least one of R1, R2, R3 and R4 includes an alkyl group —CnH2n+1 in which n is greater than or equal to 7; (b) at least two of R1, R2, R3 and R4 independently include an alkyl group —CnH2n+1 or a cycloalkyl group —CnH2n−1 in which n is greater than or equal to 4; or (c) at least three of R1, R2, R3 and R4 independently include a C1-12 alkyl group substituted by —NR5R6 or aryl.

Core Innovation

The invention relates to triazine and pyrimidine protein-binding ligand compounds of general formula (I) having defined substituents Q1 and Q2 and an attachment point A to a support matrix. Q1 represents –NR1R3 and Q2 represents –NR2R4, with R1 and R2 selected from specified alkyl groups and R3 and R4 both representing methyl. The attachment point A provides a linkage to the support matrix, optionally through a spacer linkage interposed between the matrix and the compound of formula (I).

The described compounds include hydrophobic groups in the Q1 and/or Q2 substituents, where at least one, two, or three of R1–R4 contain long-chain (C7+) alkyl or cycloalkyl (C4+) hydrophobes, including multiple substituted C1–C12 alkyl or aryl. The disclosure characterizes improved selectivity versus conventional hydrophobic interaction chromatography (HIC) ligands by using “locked” hydrophobic groups and hydrophobic pockets on the ligand. Preferred triazine embodiments and the defined substituent sets are presented for the ligand compounds.

The disclosure further describes how the ligands are provided as solid-phase ligands attached to a support matrix, including a compound library format attached at the point A to a common support matrix. A broad support-matrix concept is described, with example materials including agarose and other polymers, with an optional spacer linkage. Synthetic routes are outlined using chloro-triazine intermediates followed by coupling to install the Q2 groups, and then attachment context is established at the support matrix via the attachment point A.

The invention uses the resulting compound-ligand structures for protein binding in analytical and separation contexts. The disclosed uses include ligand use in column chromatography or micro-column/array screening to isolate, purify, or discover proteins, with examples including IL-18 binding protein. Example solid-phase ligand synthesis and screening and binding/purification results are described for IL-18 binding protein and other proteins such as ribonuclease A and human IgG.

Claims Coverage

The independent claim covers a compound of formula (I) with specified Q1 and Q2 structures, defined substituent selections for R1–R4, and an attachment point A to a support matrix, optionally via a spacer linkage. Further claim coverage includes specified attachment contexts, synthetic sequences, a library attached to a common matrix, and a process for separating, purifying, or discovering proteinaceous material using the compounds.

The core protection is directed to formula (I) protein-binding ligand compounds with Q1 and Q2 as defined, substituent selections for R1–R2, R3 and R4 as methyl, and attachment at A to a support matrix optionally via a spacer linkage. The claims further cover specified attachment contexts, synthetic sequences, a library attached to a common matrix, and use of formula (I) compounds to separate, purify, or discover proteinaceous material.

Stated Advantages

Documented Applications