The Role of High Speed Synthesis in Protein Kinase Approaches Graham F. Smith Library Design and production Group PGRD Sandwich UK

Kinases - the opportunity There are “433” kinases roles in many diseases known pharmacophores known structure motifs - crystal & homology large protein ligands common ligand ATP Weak unselective actives seem common Potency, selectivity and novelty efficiently generated are the key LD POP goals

There are “433” kinases

roles in many diseases

known pharmacophores

known structure motifs - crystal & homology

large protein ligands

common ligand ATP

Weak unselective actives seem common

Potency, selectivity and novelty efficiently generated are the key LD POP goals

High Speed Synthesis - the opportunity Many of the known pharmacophores fall within existing protocols Closed Loop process Protocol validation Diverse / targeted subsets Dedicated production HT Screen sequences iterative sequence IT & communication = oil Design / optimise VL Synthesise Screen Select subset

Many of the known pharmacophores fall within existing protocols

Closed Loop process

Protocol validation

Diverse / targeted subsets

Dedicated production

HT Screen sequences

iterative sequence

IT & communication = oil

LDP-Sandwich ~ 20k compounds per annum Sparse / diverse / targeted selections Rule of 5 compliant HPLC purified QC on every compound File enrichment stage completed ~ 42k compounds registered ArQule available for HTS large VL chemical opportunity Closed loop optimisation possible

~ 20k compounds per annum

Sparse / diverse / targeted selections

Rule of 5 compliant

HPLC purified

QC on every compound

File enrichment stage completed

~ 42k compounds registered

ArQule

available for HTS

large VL chemical opportunity

Closed loop optimisation possible

IKK2 - HTS to LD POP Simple solution phase chemistry PTFE 96 well plates Large available monomer sets

Simple solution phase chemistry

PTFE 96 well plates

Large available monomer sets

IKK HTS - 1999 UK-374,974 (10.6 µM) UK-379,552 (6.4µM) 4880 LDP kinase targeted compounds were present in file delivered a few weak actives from LDP libraries and UK-14,403 UK-379,421 (9.4 µM) UK-14,403 (ca. 2 µM) LDP libraries

4880 LDP kinase targeted compounds were present in file

delivered a few weak actives from LDP libraries and UK-14,403

IKK2 closed loops

IKK2 - summary HTS produced weak actives from LDP compounds poor leads and poor SAR closed loop work took place 6 loops performed new monomers added to VL by design after each loop VL grew from ~300K to ~4.6 Million 2.5k compounds made and screened ~0.5 Chem FTE to complete this work TA started lead development project LD-POP 3 structural series - low nM IC50’s clear SAR whole cell activity in PBMC’s

HTS produced weak actives from LDP compounds

poor leads and poor SAR

closed loop work took place

6 loops performed

new monomers added to VL by design after each loop

VL grew from ~300K to ~4.6 Million

2.5k compounds made and screened

~0.5 Chem FTE to complete this work

TA started lead development project

LD-POP

3 structural series - low nM IC50’s

clear SAR

whole cell activity in PBMC’s

Pyrazoles in cdk2 - Jay Pandit

Kin28 - the next LD POP? MISD target selection/validation Evotec HTS assay development Targeted subset pre HTS LDP compounds and IKK2 actives found cdk2 structure + chimera available legacy SAR from IKK2 Fungal cidality and selectivity are the key LD-POP goals

MISD target selection/validation

Evotec HTS assay development

Targeted subset pre HTS

LDP compounds and IKK2 actives found

cdk2 structure + chimera available

legacy SAR from IKK2

Fungal cidality and selectivity are the key LD-POP goals

Kin28 1st loop design selected IKK2 and cdk2 compounds avoid pyrazole-quinazolines - PGP? library screened in both IKK2 and Kin28 assays in parallel positional scan monomers from DISCUS Amino alcohols pyrazoles and polar hets Status…?

selected IKK2 and cdk2 compounds

avoid pyrazole-quinazolines - PGP?

library screened in both IKK2 and Kin28 assays in parallel

positional scan

monomers from DISCUS

Amino alcohols

pyrazoles and polar hets

Status…?

Other kinase hits Syk PILS Kin28 - ongoing Cdk2 - selectivity screen Lck - ? CamKII other closed loop gene families PDE2, 5, 9, 10 & 11 NPFF, mGluR1, MAdCAM, 5HT4

Syk

PILS

Kin28 - ongoing

Cdk2 - selectivity screen

Lck - ?

CamKII

other closed loop gene families

PDE2, 5, 9, 10 & 11

NPFF, mGluR1, MAdCAM, 5HT4

monomers protocols + = select synthesise screen PGRD core assets developing competencies expand & beautify apply // synthesis to generate LD-POPs Shared VL approach to kinases? AIDD / LiBrain parallel chemistry space

Thanks to... Mark Gardner Wolfgang Klute Nunzio Sciammetta Dave Williams Snahel Patel Mark Kemp Duncan Armour Sandy Monaghan Martin Edwards Pam Greengrass Sue Boughton John Taylor Steve Liu Mike Yeadon Kate Burt Colin Groom Rachel Grimley Blanda Stammen Tanya Parkinson

Mark Gardner

Wolfgang Klute

Nunzio Sciammetta

Dave Williams

Snahel Patel

Mark Kemp

Duncan Armour

Sandy Monaghan

Martin Edwards

Pam Greengrass

Sue Boughton

John Taylor

Steve Liu

Mike Yeadon

Kate Burt

Colin Groom

Rachel Grimley

Blanda Stammen

Tanya Parkinson