copyright © 2007 merck & co., inc., whitehouse station, new jersey, usa all rights reserved 14...

Copyright © 2007 Merck & Co., Inc., Whitehouse Station, New Jersey, USA All rights Reserved

14th CROIFebruary 27, 2007

Next Generation Inhibitors of HIV-1 Integrase Strand Transfer:Structural Diversity and Resistance Profiles

John WaiMerck Research Laboratories West Point, PA

2 Copyright © 2007 Merck & Co., Inc., Whitehouse Station, New Jersey, USA All rights Reserved

HIV Life Cycle


N N

HN

O

OHO

HN

O

NN

O

F

• Potent integrase strand transfer inhibitor (IC50 ~ 10 nM)• Antiviral IC95 ~ 33 23 nM (50% NHS)• Retains potency across diverse clinical isolates, HIV-2• Potent antiviral effect in treatment naïve patients (Markowitz et al IAC 2006)• Robust activity in HIV-1 patients with triple class resistance

– Phase II 24 week data (Grinsztejn et al CROI 2006)– Phase III data (Cooper et al & Steigbigel et al CROI 2007)

•Lipid neutral in naïve patients (Mascolini et al ICAAC 2006)

• Few drug interactions; Metabolized via glucuronidation

MK-0518: Raltegravir


MAINTAIN favorable properties of Raltegravir with respect to

• efficacy, • tolerability, lipid profile • few drug interactions

and ACHIEVE

• Once daily dosing (stand alone)• Activity against InSTI resistant isolates

Question: Clinical resistance profile of InSTIs???

Objectives for 2nd generation InSTI


E152

D64 D116

Mg

In Vitro Selection for Mutants

Catalytic Residues


S153

I151

F121

T125V72

M154

N155

T66

E152

D64 D116

Mg


Catalytic Residues

Diketoacid

Naphthyridine L-810


Q148

S153

I151

F121

T125V72

M154

N155

T66

E152

D64 D116

G140 E138

Mg


• Bind to catalytic site

• Different inhibitors selected for different sets of mutants

• Multiple possible binding orientations

• Mutation points are spread around the catalytic site

Catalytic Residues

Diketoacid

Naphthyridine L-810

Pyrimidine MK-518


Enabling Tool: Mutant Panel

Q148

S153

F121

T125V72

N155

T66

G140 E138

Mg

• Mutant viruses constructed using site directed mutagenesis

• Activity assessed with single cycle infectivity assay

Catalytic Residues

Diketoacid

Naphthyridine L-810

Pyrimidine MK-518


N

OHO

N N N

O

OHO

N

N X

OH

O

OHO

N

N

O

X

O

OEt

OH

X = CH, NH

X = CH, NHN

N

OHO

NH

O

N

N

O

N

OH

N

O

C2 symmetry

Diversification of DKA Isosteres

O

OH

OHO

DiketoacidPharmacophore

N

O

N N

OHO


Optimization Against InSTI Mutants

Spread

T66I/S153Y F121YQ148K

N

O

F

N N

OHO

156 22x 76x 20x31x

ClC95 nM

90x

Ratio of IC50’s inInfectivity assay N155S

E138A/G140A/Q148K

N

N

O

F

N N

OHO

Cl 9 1x 2x 1x2x

OMeHN

16x

0.8 1x 1x 1x1xN

N

O

F

N N

OHO

Cl

N

N

SO

O

1x

N

O

N N

OHO

MK-2048

Benchmark


T66I

/S15

3Y

N15

5S

Q14

8KE1

38A

/G14

0A/Q

148K

F121

YMK-2048

MK-518

GS-91370

20

40

60

80

100116X

169X

395X

388X

IC50

rat

io o

f m

uta

nts

vs.

WT

HIV

-1

in in

fect

ivit

y as

say

Comparison with Clinical CandidatesScreening Mutant Panel


Comparison with Clinical CandidatesMK-518 Clinical Isolates

IC50

rat

io o

f m

uta

nts

vs.

WT

HIV

-1

in in

fect

ivit

y as

say

N15

5HE92

Q/N

155H

Q14

8RG

140S

/Q14

8HG

140S

/Q14

8RMK-2048

MK-518

GS-91370

100

200

300399 / 2194X


WeeksPrimary Mutation

• Selection of Resistance in cell culture at IC95

• Time taken for the first emergence of mutant in vitro correlates with mutation profiles of inhibitors

Time to Selection of Resistance inCell Culture

7

11

21

>37

N155H

Q148K

S153Y

T112I

Compound B

MK-518

Compound A

Nu

mb

ers

of

wee

ks o

f in

cub

atio

n f

or

the

firs

t em

erg

ence

of

mu

tati

on

in a

ll is

ola

tes

# of Clones

5/5

6/6

7/7

5/12MK-2048

>37 wk

0

10

20

30

40 >37 wk


Summary: Second Generation InSTIs

• Resistance profiling leads to the identification of inhibitors with activity against broad spectrum of integrase resistance isolates.

• Provides Proof of Concept for developing second generation InSTIs.

T66I

/S15

3Y

N15

5S

Q14

8K

E138

A/

G14

0A/

Q14

8K

F121

Y

MK-2048

MK-518

GS-91370

20

40

60

80

100116X

169X

395X

388XIC

50 r

ati

o o

f m

uta

nts

vs.

WT

HIV

-1

in i

nfe

ctiv

ity

as

say


2nd Generation HIV-1 InSTIs Discovery Team

Medicinal ChemistryThor FisherMark EmbreyBo-young KimMelissa EgbertsonWei HanDebbie PerlowRowena RuzekPeter WilliamsMarie LangfordJanetta PellicoreDonnette StaasVanessa ShermanKelly SavageCathy WiscountLee TranMelody McWherterLinghang ZhuangKyle RobinsonMatt MorrissetteLou Anne NeilsonRichard IsaacsLinda PayneRick PracittoVanessa ObligadoWayne ThompsonPeter MunsonBrian PhillipsJohn WaiTerry LyleJoe Vacca

Biological ChemistryDaria HazudaMike MillerJay GroblerPete FelockKara StillmockAmy SimcoeMarc WitmerAlysha DayRobert DanovichYuxiong Ke

Viral Vaccine ResearchJessica FlynnLinda EctoSinouen TouchLori GabryelskiBill Schleif

Drug MetabolismChuck ThompsonPrasad KariJoan EllisKim MichelAnne TaylorReza AnariChris Kochansky

Pharm R&DDave DubostWei Xu

Scale-up LabJohn SwestockRick WoodwardRandy NewtonKim StraussJim Guare

NMR & MS GroupsSteve PitzenbergerSandor VargaMike BoguskyJanine Brouillette Chuck RossJoan Murphy

Molecular ModelingKate HollowayChris Culberson

Chromatography GroupCarl HomnickTodd AndersonChristy Olson

Analytical GroupKristi HoffmanKen AndersonMatt ZradaAmanda CortesDeanne Rudd

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