Irreversible Inhibitors of ErbB-family of kinasesGlaxoSmithKline (2002-2004)Robert D Hubbard, Ph.D.

GlaxoSmithKline (2002-2004)Irreversible Inhibitors of ErbB-family

N

N

HN

O

NHS

OO Cl

OF

TykerbTM

Reversible EGFR/ ErbB-2BT474 Proliferation (IC50,nM) = 100

N

NS

HN

NH

HCl

OF

Irreversible EGFR/ ErbB-2EGFR modification @ 3hr = 47%

BT474 Proliferation (IC50,nM) = 94

Explore the SAR on substituted pyrrolidines: modification, cellular activityPNAS 2008, 105,2773

N

NS

HN Cl

OF

H

SCys797 Structure in ErbB-4

A. Waterson, D. Uehling.

Proposed mechanism of modification requires a basic pyrrolidine crucial

Analog BT474 (IC50, nM)

EGFR mod. @ 3 hr (%)

R = H 94 47R = -(CH2)2OMe

110 19

R = -SO2Me 650 0R = -C(O)OMe 1700 0 BMCL 2008, 18, 5738

N

NS

HN

N

HCl

OF

R

TykerbTM (green)Thienopyrimidine (red)

Substituted Carbamoyl-pyrrolidinesProvided a balance of activity and minimized EGFR modification

N

NS

HN

NH

Cl

OF

N

NS

HN

NH

Cl

OF

OHN

O

N

NS

HN

NH

Cl

OF

ON

O

N

NS

HN

NH

Cl

OF

ON

O

O

ON

O

R1

R2

N

NS

HN

BrN

Cl

OF

ON

O

R1

R2

N

HO

BOC1. NaH, then R1R2COCl

2. Sonogashira Coupling3. TFA

BOC

CO2MeN

HO

BOC

Corey-Fuchs

BMCL 2009, 19, 21.

SAR comments: Potency: Et > Dimethyl > MorpholinePK: Morpholine ~ Dimethyl > EtModification of EGFR @ 3hr: < 15%

From Laulimalide total synthesis (Stanford)Modified Seyferth-Gilbert Reaction

OH

HO

OTBS

OTBSO

H

1. MOMCl, DIEA, CH2Cl2, RT (99%)2. (a) BH3-DMS, cyclohexene, THF(b) H2O2, 3 M NaOH, EtOH (87%)

3. Dess-Martin, NaHCO3, CH2Cl2, H2O (85%)O

H

MOMO

OTBS

OTBSO

H

O

4. K2CO3, MeOH (76%)

5. n-BuLi, ClCO2Me, THF,-78C (79%, 86% BORSM)

OH

MOMO

OTBS

OTBSO

H

CO2Me

6. HF-pyridine, THF, RT (98%)

7. LiOH, THF, water, RT (76%)O

H

MOMO

OH

OHO

H

CO2H

8. 2,4,6-trichlorobenzoyl chloride,Et3N, DMAP, benzene, RT (55%)

O

O

OHO

H

OMOMH

OH

O

OO

OHHHO

H

OHO

H

Laulimalide

O

N2

PO

O

O

JACS 2002, 124, 4956.

Synthesis of all possible diastereomers (4-OH proline)

N

HO

CO2Me

BOC

1. TBDPSCl, Imid2. LiBH4, THF

3. Dess-Martin, CH2Cl2,water

N

TBDPSO

CHO

BOC

P

N2

OO

K2CO3, MeOH, RT N

TBDPSO

BOC

4. 5. TBAF, THF

(R)

N(S)

HO

BOC

7. Ph3P, DIAD, pNO2-PhCO2H

(S)

N(S)

HO

BOC

O O

NH

HO

CO2Me1. Ac2O, AcOH

NO

ORR = -Ac

then HCl, refluxNH

HO

CO2H2. SOCl2, MeOH

3. BOC2, TEA, DMAP N

HO

CO2Me

BOC

4. TBDPSCl, Imid

5. DIBAL-H, -78C, then MeOH

N

TBDPSO

CHO

BOC

OP

N2

OO

O

N

TBDPSO

BOC

6. TBAF, THF(R)

N(R)

HO

BOC

7. Ph3P, DIAD, pNO2-PhCO2H

(S)

N(R)

HO

BOC

BMCL 2009, 19, 21.

SAR analysis of four possible stereoisomers of the pyrrolidinyl-carbamate – SAR consistent for all carbamates

Analog

BT474 (IC50, nM) 30 50 130 10

Ms PO PK (mM*hr) 1.5 ~ 0.3-0.5 low low

N

NS

HN

R

Cl

OF

NH

ON

O NH

ON

ONH

ON

ONH

ON

O

N CO2Me

HO

BOC

(-)-Aldrich

N CO2Me

HO

BOC(+)-not available, crazy expensive

Letter from Aldrich??

GSK – Summary

• Decoupled EGFR modification from cellular potency

• SAR of cellular activity, PK, and modification dependent on the stereochemistry of the substituents on the pyrrolidine ring

• Project was terminated due to concerns of irreversible inhibition of target kinases.

• T790M mutation requires irreversible inhibition for potent activity (PNAS 2008, 105, 2070).