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Design of Experiments Design of Experiments (DOE): (DOE): A “New” Approach to A “New” Approach to Reaction Optimization Reaction Optimization Dr. Steven Weissman Dr. Steven Weissman Merck & Co. Merck & Co. Feb 4, 2008/UPR Feb 4, 2008/UPR
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DOE Applications in Process Chemistry Presentation

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Page 1: DOE Applications in Process Chemistry Presentation

Design of Experiments (DOE): Design of Experiments (DOE): A “New” Approach to Reaction A “New” Approach to Reaction

OptimizationOptimization

Dr. Steven WeissmanDr. Steven WeissmanMerck & Co. Merck & Co.

Feb 4, 2008/UPR Feb 4, 2008/UPR

Page 2: DOE Applications in Process Chemistry Presentation

OutlineOutline

Background: Big changes for Pharma Background: Big changes for Pharma

Basic Principles of Basic Principles of Design of ExperimentsDesign of Experiments

Merck Case StudiesMerck Case Studies

Take Home message/QuestionsTake Home message/Questions

Page 3: DOE Applications in Process Chemistry Presentation

Big Changes for Big PharmaBig Changes for Big Pharma

Costs/Risks of drug development are risingCosts/Risks of drug development are rising– low hanging fruit has been pickedlow hanging fruit has been picked– small molecules no longer in voguesmall molecules no longer in vogue

protein-based and vaccines = more opportunitiesprotein-based and vaccines = more opportunities

– ‘‘Vioxx hangover’- more trials/more patients = $$Vioxx hangover’- more trials/more patients = $$

Page 4: DOE Applications in Process Chemistry Presentation

Big Changes for Big PharmaBig Changes for Big Pharma

Costs/Risks of drug development are risingCosts/Risks of drug development are rising

Globalization of marketplaceGlobalization of marketplace– US market sales is matured; slow growthUS market sales is matured; slow growth– Emerging markets sales = high growth potentialEmerging markets sales = high growth potential– Strategic portions of drug development Strategic portions of drug development

outsourced to India/China (low-cost providers)outsourced to India/China (low-cost providers)

Page 5: DOE Applications in Process Chemistry Presentation

Big Changes for Big PharmaBig Changes for Big Pharma

Costs/Risks of drug development are risingCosts/Risks of drug development are rising

Globalization of marketplaceGlobalization of marketplace

Uncertain pipelinesUncertain pipelines– ‘‘Batting average’ is unchanged despite huge Batting average’ is unchanged despite huge

investments investments – Is bureaucracy killing drug discovery?Is bureaucracy killing drug discovery?

Are smaller companies becoming better at this?Are smaller companies becoming better at this?

Page 6: DOE Applications in Process Chemistry Presentation

Big Changes for Big PharmaBig Changes for Big Pharma

Costs/Risks of drug development are risingCosts/Risks of drug development are risingGlobalization of marketplaceGlobalization of marketplaceUncertain pipelinesUncertain pipelinesRevenues/profits are being squeezedRevenues/profits are being squeezed– patent expirations – Fosamax ($3 B)patent expirations – Fosamax ($3 B)– tougher regulatory environmenttougher regulatory environment– payers demand value-addedpayers demand value-added– lower cost structures: aggressively pursuedlower cost structures: aggressively pursued

downsizingdownsizingplant closings-namely here in PRplant closings-namely here in PR

Page 7: DOE Applications in Process Chemistry Presentation

Big Changes for Big PharmaBig Changes for Big Pharma

Costs/Risks of drug development are risingCosts/Risks of drug development are rising

Globalization of marketplaceGlobalization of marketplace

Uncertain pipelinesUncertain pipelines

Revenues/profits are being squeezedRevenues/profits are being squeezed

Page 8: DOE Applications in Process Chemistry Presentation

New Approaches NeededNew Approaches Needed

What can we do as chemists to What can we do as chemists to change the way we do our jobs ?change the way we do our jobs ?

Can we work smarter/faster ?Can we work smarter/faster ?

How ?? How ??

Page 9: DOE Applications in Process Chemistry Presentation

New Approaches NeededNew Approaches Needed

What can we do as chemists to What can we do as chemists to change the way we do our jobs ?change the way we do our jobs ?

Can we work smarter/faster ?Can we work smarter/faster ?

How ?? How ?? Automation/TechnologyAutomation/Technology

Page 10: DOE Applications in Process Chemistry Presentation

Automated Synthesis CycleAutomated Synthesis Cycle

Design

Experiment Analysis

Informatics

Page 11: DOE Applications in Process Chemistry Presentation

Automated Synthesis CycleAutomated Synthesis Cycle

Designof Experiments

Design

Experiment Analysis

Informatics

Page 12: DOE Applications in Process Chemistry Presentation

Current Approach to OptimizationCurrent Approach to Optimization

Change One Factor at a time (OFAT)Change One Factor at a time (OFAT)

– Rarely leads to optimal conditionsRarely leads to optimal conditions– Leads to different conclusions depending on Leads to different conclusions depending on

starting pointstarting point– Requires many expts/little informationRequires many expts/little information– Cannot separate “noise” from true variabilityCannot separate “noise” from true variability

Page 13: DOE Applications in Process Chemistry Presentation

Current Approach to OptimizationCurrent Approach to Optimization

Change One Factor at a time (OFAT)Change One Factor at a time (OFAT)

– Rarely leads to optimal conditionsRarely leads to optimal conditions– Leads to different conclusions depending on Leads to different conclusions depending on

starting pointstarting point– Requires many expts/little informationRequires many expts/little information– Cannot separate “noise” from true variabilityCannot separate “noise” from true variability

– Ignores interactions of variablesIgnores interactions of variables

Page 14: DOE Applications in Process Chemistry Presentation

Example of OFAT (11/07)Example of OFAT (11/07)

Page 15: DOE Applications in Process Chemistry Presentation

21 Reactions21 Reactions

Page 16: DOE Applications in Process Chemistry Presentation

DOE vs OFATDOE vs OFAT

OFAT: 3 factors needed 21 reactionsOFAT: 3 factors needed 21 reactions– No information on interactions of effectsNo information on interactions of effects

DOE: 3 or 4 factors- 11 or 17 reactionsDOE: 3 or 4 factors- 11 or 17 reactions– Better quality informationBetter quality information– Learn about interactions of effectsLearn about interactions of effects– Fewer reactionsFewer reactions

Page 17: DOE Applications in Process Chemistry Presentation

Notable QuoteNotable Quote

““If you test one factor at a time, there’s a If you test one factor at a time, there’s a low probability that you are going to hit low probability that you are going to hit the right one before everybody gets sick the right one before everybody gets sick of it and quits”of it and quits”

Forbes magazine article on DOE in 1996Forbes magazine article on DOE in 1996

Page 18: DOE Applications in Process Chemistry Presentation

What is DOE ?What is DOE ?

Selected set of expts in which all relevant Selected set of expts in which all relevant factors are varied factors are varied simultaneouslysimultaneously

‘‘Continuous’ factors are ideal (time, temp, Continuous’ factors are ideal (time, temp, equiv)-the ‘equiv)-the ‘How much’How much’

Analysis reveals which factors influence the Analysis reveals which factors influence the outcome and identifies optimal conditionsoutcome and identifies optimal conditions

Systematic, organized approach to problem Systematic, organized approach to problem solvingsolving

Mathematical model of the design spaceMathematical model of the design space

Page 19: DOE Applications in Process Chemistry Presentation

DoE IntroductionDoE Introduction

Core Knowledge(Engineering, Chemistry, Biology,…)

Statistical Knowledge

Develop Solutions

DOE is NOT a replacement for process knowledge

Page 20: DOE Applications in Process Chemistry Presentation

Questions to be Answered by DoEQuestions to be Answered by DoE

How do we get the best synthetic yield ?How do we get the best synthetic yield ?

How much catalyst/ligand do I need ?How much catalyst/ligand do I need ?

Can we minimize formation of an impurity?Can we minimize formation of an impurity?

Which experimental factors are (un) Which experimental factors are (un) important?important?

How robust is my process ?How robust is my process ?

Page 21: DOE Applications in Process Chemistry Presentation

DOE: ConsiderationsDOE: Considerations

Can’t replace full screening of catalyst or Can’t replace full screening of catalyst or solvent (HTS)- ‘solvent (HTS)- ‘discreetdiscreet’ variables’ variables Best suited for Best suited for continuouscontinuous variables variables– time, temp, stoichiometrytime, temp, stoichiometry

Not helpful for non-reproducible rxnsNot helpful for non-reproducible rxnsBest suited for ‘low maintenance’ rxnsBest suited for ‘low maintenance’ rxns– Temp = 20 to 150 Temp = 20 to 150 ooCC– All reactants added at once All reactants added at once

Page 22: DOE Applications in Process Chemistry Presentation

DOE: Experimental ObjectivesDOE: Experimental Objectives

ScreeningScreening– Which factors are most influential ? Which factors are most influential ? – What are their appropriate values/ranges ?What are their appropriate values/ranges ?

OptimizationOptimization– Extract information regarding how factors Extract information regarding how factors

combine to influence responsecombine to influence response– Identify optimized reaction conditionsIdentify optimized reaction conditions

Page 23: DOE Applications in Process Chemistry Presentation

DOE: MisconceptionsDOE: Misconceptions

Requires in-depth statistics knowledgeRequires in-depth statistics knowledge– User-friendly DOE software does this for youUser-friendly DOE software does this for you

MODDEMODDE (Umetrics)/ (Umetrics)/Design ExpertDesign Expert (Stat-ease) (Stat-ease)

Page 24: DOE Applications in Process Chemistry Presentation

DOE: MisconceptionsDOE: Misconceptions

DoE requires in-depth statistics knowledgeDoE requires in-depth statistics knowledge– Experimental design software does this for youExperimental design software does this for you

DoE requires a lot of experiments and timeDoE requires a lot of experiments and time– Perhaps. but will always get better quality informationPerhaps. but will always get better quality information– Typically 11-27 reactions per designTypically 11-27 reactions per design– Automation/technology helps reduce the effort neededAutomation/technology helps reduce the effort needed

Page 25: DOE Applications in Process Chemistry Presentation

High Throughput ScreeningHigh Throughput Screening

= 96 x

Discreet variables- ‘The what”

What is the best ligand/catalyst combination ?What is the best solvent ?

Page 26: DOE Applications in Process Chemistry Presentation

High Throughput ScreeningHigh Throughput Screening

= 96 x

Can we do OPTIMIZATION this way too ??

Page 27: DOE Applications in Process Chemistry Presentation

High Throughput Optimization ??High Throughput Optimization ??

= 96 x

If so,…………………..Which reactions do we run ?How do assess the data ?

Page 28: DOE Applications in Process Chemistry Presentation

High Throughput Optimization ??High Throughput Optimization ??

= 96 x

Statistical Design of Experiments (DOE)

Page 29: DOE Applications in Process Chemistry Presentation

HTS Reaction VialsHTS Reaction Vials

Page 30: DOE Applications in Process Chemistry Presentation

DOE: WorkflowDOE: WorkflowDefine the Define the Objective Objective – screening, optimize, robustnessscreening, optimize, robustnessDefinition of Definition of Factors Factors – Prioritize: known, suspected, possibly, unlikelyPrioritize: known, suspected, possibly, unlikely– Set HIGH/LOW values for factors (define Set HIGH/LOW values for factors (define design design

spacespace))Define the Define the Response Response – how to measure ?– how to measure ?Select Select Experimental DesignExperimental Design Generate Generate WorksheetWorksheet Run theRun the Reactions Reactions PerformPerform Analysis Analysis with DOE softwarewith DOE software

Page 31: DOE Applications in Process Chemistry Presentation

DOE Design (N=27)DOE Design (N=27)Factor 1 Factor 2 Factor 3 Factor 4 Factor 5 Response 1

Rxn # A:temp B:P/Pd C:Cu D:Boron/Br E:Conc Yield

1 85 2 0.25 1.2 0.4 45

2 115 2 0.25 1.2 0.1 49

3 85 4 0.25 1.2 0.1 43

4 115 4 0.25 1.2 0.4 55

5 85 2 1.5 1.2 0.1 45

6 115 2 1.5 1.2 0.4 55

7 85 4 1.5 1.2 0.4 39

8 115 4 1.5 1.2 0.1 54

9 85 2 0.25 2.5 0.1 48

10 115 2 0.25 2.5 0.4 63

11 85 4 0.25 2.5 0.4 88

12 115 4 0.25 2.5 0.1 76

13 85 2 1.5 2.5 0.4 79

14 115 2 1.5 2.5 0.1 65

15 85 4 1.5 2.5 0.1 80

16 115 4 1.5 2.5 0.4 76

17 85 3 0.875 1.85 0.25 64

18 115 3 0.875 1.85 0.25 66

19 100 2 0.875 1.85 0.25 58

20 100 4 0.875 1.85 0.25 64

Page 32: DOE Applications in Process Chemistry Presentation

DOE Creates a Design SpaceDOE Creates a Design Space

Design-Expert® Software

YieldX1 = A: tempX2 = B: P/PdX3 = C: Cu load

Actual FactorsD: Boron/Br = 2.50E: Conc = 0.40

CubeYield

A: temp

B:

P/P

d

C: Cu load

A-: 85.00 A+: 115.00B-: 2.00

B+: 4.00

C-: 0.25

C+: 1.50

63.7936

74.1825

86.3492

83.738

59.9047

70.2936

82.4603

79.8492

Page 33: DOE Applications in Process Chemistry Presentation

DOE Expts: How Many ?DOE Expts: How Many ? rxnsrxns

factorsfactors

HIHI Med Med LoLo Total rxnsTotal rxns

3 factors3 factors 44 33 44 1111

44 88 33 88 1919

5 5 1616 33 1616 3535

33 55 77 55 1717

44 1010 77 1010 2727

55 99 1111 99 2929

scree

nin

g o

ptim

aztio

n

Page 34: DOE Applications in Process Chemistry Presentation

DOE Case StudiesDOE Case Studies

Page 35: DOE Applications in Process Chemistry Presentation

MK-0518MK-0518

N

N

O

OK

O

HN

HN

O

NN

O

F

First-in-Class Oral HIV-1 Integrase Inhibitor

Approved by FDA October-12-2007

Page 36: DOE Applications in Process Chemistry Presentation

MK-0518MK-0518

N

N

O

OH

O

HN

F

HN

O

NN

O

N

N

O

OH

O

H2N OMe

H2N

F

O

NN

OOK

Page 37: DOE Applications in Process Chemistry Presentation

MK-0518MK-0518

N

N

O

OH

O

HN

F

HN

O

NN

O

N

N

O

OH

O

H2N

F

O

NN

OOK

HN

Challenge: to reduce manufacture cost by 20%

Page 38: DOE Applications in Process Chemistry Presentation

MK-518: Problem stepMK-518: Problem step

Peter Maligres

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

DMSO

58 C/5 h

Existing Conditions: 4 eq Mg(OMe)2/ 4 eq MeI @ 0.5 M (68% isolated yield)

18 solvents, 8 bases screened

78 22

Page 39: DOE Applications in Process Chemistry Presentation

MK-518: DOE OptimizationMK-518: DOE Optimization

Peter Maligres

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

DMSO

DOE Optimzation Design Factors: Mg(OMe)2 equiv: 1.0 and 3.0MeI equiv: 2.5 and 5.0Conc: 0.25 and 1.0 MTemperature: 30 and 65 oC

19 reactions

Responses (4 and 20 h):Assay yieldSelectivity

Page 40: DOE Applications in Process Chemistry Presentation

MK-518 OptimizationMK-518 Optimization

Peter Maligres

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

DMSO

20 h

> 90% AY

DOE Optimal SettingsBase equiv: 1.0 and 3.0 MeI equiv: 2.5 and 5.0Temperature: 30 and 65 oCConc: 0.25 and 1.0 MTime: 4 and 20 h

99 1

Page 41: DOE Applications in Process Chemistry Presentation

MK 518: Surface ModelMK 518: Surface ModelInvestigation: MK 518 Methylation 20 hr AB ratio (MLR)

Response Surface Plot

MODDE 8 - 10/26/2007 4:23:15 PM

Base Equiv = 3Temp = 65

Page 42: DOE Applications in Process Chemistry Presentation

Effect of Temp & ConcEffect of Temp & ConcInvestigation: MK 518 Methylation 20 hr AB ratio (MLR)

Contour Plot

MODDE 8 - 12/13/2007 11:25:45 AM

Base Equiv = 3MeI equiv = 5

Page 43: DOE Applications in Process Chemistry Presentation

Effect of Base and ConcEffect of Base and ConcInvestigation: MK 518 Methylation 20 hr AB ratio (MLR)

Contour Plot

MODDE 8 - 12/13/2007 11:38:40 AM

MeI equiv = 5Temp = 65

Page 44: DOE Applications in Process Chemistry Presentation

MK518-In Situ DemethylationMK518-In Situ Demethylation

HN

N

O

OH

O

CbzHNHN

N

N

O

OH

O

CbzHNHN

F F

Mg(OMe)2then MeIDMSO N

N

OMe

OH

O

CbzHNHN

F

+

Magnesium iodide salts

4 h4 h 20 h20 h

ConvConv 95%95% 99%99%

N vs ON vs O 80/2080/20 99/199/1

Page 45: DOE Applications in Process Chemistry Presentation

MK-518 ConcernsMK-518 Concerns

Peter Maligres

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

DMSO (1M) 65 oC

3 eq Mg(OMe)25 eq MeI

Issues:

1. at this higher concentration, end of reaction difficult to stir2. Mg(OMe)2- long term issues with supply & cost3. MeI is mutagenic/carcinogen/toxic

99 1

Page 46: DOE Applications in Process Chemistry Presentation

MK-518 OptimizationMK-518 Optimization

Peter Maligres

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

NMP (1.2 M)100 oC/6 h

2 eq Mg(OH)22 eq Me3SOI

Yield =90%Selectivity = >99.9 %Safer, more economical reagentsIncorporated best practices from DOE:

HI Temp/HI Concentration/Longer reaction times

Page 47: DOE Applications in Process Chemistry Presentation

MK-518 SummaryMK-518 Summary

HN

NCbzN

OH

O

H MeN

NCbzN

OH

O

H N

NCbzN

OH

OMe

H+

HN

O

F

O

HN

O

HN

F F

> 90% Yield

78 22

> 99 < 1

DOE

Goal of 20% reduction in drug inventory cost was achievedHigher Yield cascades back to allow fewer RM/solvents to be usedSubmitted for 2008 Presidential Green Chemistry Award

Page 48: DOE Applications in Process Chemistry Presentation

Case Study 2- SuzukiCase Study 2- Suzuki

Br R

R

(HO)2B

Tol/ THF, K2CO3

Pd(OAc)2/Ar-PCy2

95% AY92 A% pure

70 oC

Goal: 1. to reduce Pd(OAc)2 & ligand charges (0.4 mole%,0.8 mole%)2. Improve yield and/or purity

Page 49: DOE Applications in Process Chemistry Presentation

Case Study 2- SuzukiCase Study 2- Suzuki

DOE Factors:

Ligand/Pd ratio: 1.0 and 3.0Catalyst load: 0.1 and 0.5 mole%Molarity boronic acid: 0.5 and 1.5Temperature: 60 and 80 oC

27 Reactions in 96-well plate format, 2 days to plan/setup/execute/assay0.65 g material (24 mg/rxn) !!

Br R

R

(HO)2B

Tol/ THF, K2CO3

Pd(OAc)2/Ar-PCy2

95% AY92 A% pure

70 oC

Page 50: DOE Applications in Process Chemistry Presentation

Case Study 2- SuzukiCase Study 2- Suzuki

DOE Optimal Settings:Ligand/Pd ratio: 1.0 and 3.0Catalyst load: 0.1 and 0.5 mole%Molarity boronic acid : 0.5 and 1.5Temperature: 60 and 80 oC (65 oC)

Br R

R

(HO)2B

Tol/ THF, K2CO3

Pd(OAc)2/Ar-PCy2

Page 51: DOE Applications in Process Chemistry Presentation

Effect of Temp and Pd LoadingEffect of Temp and Pd Loading

Lig/ catalyst ratio fixed at 3:1; Triol M fixed at 1.5 M

Overall LCAP

MODDE 8 - 1/8/2008 5:20:14 PM

L i g / C a t = 3

Page 52: DOE Applications in Process Chemistry Presentation

OptimizedOptimized Conditions Conditions

Optimized Experiment:-increased LCAP by 1%-decreased DesBr impurity (50%)-decreased Pd by 75%-decreased Lig by 70%

Spencer Dreher

BrR

R

(HO)2B

Tol/ THF, K2CO3

0.1 %Pd(OAc)2

95% AY93 A% pure

70 oC

0.25 % Ar-PCy2

1.5 M

Page 53: DOE Applications in Process Chemistry Presentation

Case Study #3Case Study #3

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

> 99% conv>98% ee

Dave Pollard

Goal: to reduce cost by increasing productivity

100 g/L

Page 54: DOE Applications in Process Chemistry Presentation

Screening DesignScreening Design

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

Dave Pollard

FactorsOctanol: 40 and 60 %NADP equiv: 0.1 and 0.5 %Concentration: 50 and 150 g/LTemp: 25 and 35 oCEnzyme load: 0.3 to 1.0 g/L

19 experiments

Page 55: DOE Applications in Process Chemistry Presentation

DOE Factors PlotDOE Factors Plot

-1.0

-0.8

-0.6

-0.4

-0.2

-0.0

0.2

0.4

0.6

0.8

NA

D

Co

n

Te

mp

en

z

NA

D*C

on

Co

n*T

em

p

%

Investigation: -Ketone Bio-Reduction-Conv2 (MLR)

MODDE 8 - 2/1/2008 12:55:49 PM

Page 56: DOE Applications in Process Chemistry Presentation

Interaction: Conc and NADInteraction: Conc and NAD

-1.4

-1.2

-1.0

-0.8

-0.6

-0.4

-0.2

0.0

0.2

0.4

0.6

0.8

50 60 70 80 90 100 110 120 130 140 150

Co

nv

ers

ion

Concentration

NAD (low )NAD (high)

NAD (low)

NAD (low)

NAD (high)

NAD (high)

MODDE 8 - 2/1/2008 12:53:16 PM

Page 57: DOE Applications in Process Chemistry Presentation

Screening ResultScreening Result

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

Dave Pollard

Factor Preferred SettingOctanol: 40 and 60 % No impactNADP equiv: 0.1 and 0.5 % No impact- increase more ? Concentration: 50 and 150 g/L 50 g/L- undesirable settingTemp: 25 and 35 oC minimal effect- set at 30 oCEnzyme load: 0.3 to 1.0 g/L 1.0 g/L-increase more

Page 58: DOE Applications in Process Chemistry Presentation

Optimization DesignOptimization Design

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

Dave Pollard

FactorNADP equiv: 0.5 and 1.5 % Concentration: 100 and 200 g/LEnzyme load: 0.5 to 3.0 g/L

19 experiments

Page 59: DOE Applications in Process Chemistry Presentation

Optimization DesignOptimization Design

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

Factor Preferred settingNADP equiv: 0.5 and 1.5 % No effect Concentration: 100 and 200 g/L 200Enzyme load: 0.5 to 3.0 g/L 3.0

Page 60: DOE Applications in Process Chemistry Presentation

Optimization DesignOptimization Design

N

OCl

N

OH

Cl

OHO

NADP+NADPH

Enzyme

2-octanol/buffer, pH 6, 30 oC

H2SO4

Factor Preferred settingNADP equiv: 0.5 and 1.5 % No effect Concentration: 100 and 200 g/L 200Enzyme load: 0.5 to 3.0 g/L 3.0

Confirming experiment at 200 g/L NADP= 0.5 g/L and enzyme at 3 g/L gave 100% conversion Goal achieved

Page 61: DOE Applications in Process Chemistry Presentation

Case Study #4-SonogashiraCase Study #4-Sonogashira

NH

O

I

15 mol% CuI2 equiv TEA

DMF25 oC/1 h

~100% conversion33% isolated yield

10 mol% Pd(TPP)4

1.2 eq

NH

O NH

O

H+ + bis-addition

impurity

S. Krska/A. Northrup

Medicinal Chemistry conditions

Page 62: DOE Applications in Process Chemistry Presentation

HTS ResultHTS Result

NH

O

I

NH

O

3

40 mol% CuI3 equiv iPr2NH

MeCN45 oC/18 h

90 A%

10 mol% [(allyl)PdCl]240 mol% (2-furyl)3P

+

10 A%

bis additionimpurity

Screened: ligands and Pd sources32 reactions (HTS-96 well plate format) – 1.5 days- 125 mg of substrate

Page 63: DOE Applications in Process Chemistry Presentation

DOE OptimizationDOE Optimization

3 Factor RSM design – 19 reactionsFixed factors: L/Pd ratio (2:1); temperature (45 oC); base equiv (3)

Varied Factors:• Pd loading (2 and 10 mole%)• Cu/Pd ratio (0.5 and 2.0)• Alkyne equiv (1 and 5)

NH

O

I

NH

O

3

40 mol% CuI3 equiv iPr2NH

MeCN45 oC/18 h

90 A%

10 mol% [(allyl)PdCl]240 mol% (2-furyl)3P

+

10 A%

bis additionimpurity

Page 64: DOE Applications in Process Chemistry Presentation

DOE OptimizationDOE Optimization

3 Factor RSM design – 19 reactionsFixed factors: L/Pd ratio (2:1); temperature (45 oC); base equiv (3)

Varied Factors:• Pd loading (2 and 10 mole%)- little effect- set to 3 mole%• Cu/Pd ratio (0.5 and 2.0)- most important therefore 12 mole% CuI• Alkyne equiv (1 and 5) –2 equiv

NH

O

I

NH

O

3

40 mol% CuI3 equiv iPr2NH

MeCN45 oC/18 h

90 A%

10 mol% [(allyl)PdCl]240 mol% (2-furyl)3P

+

10 A%

bis additionimpurity

Page 65: DOE Applications in Process Chemistry Presentation

DOE OptimizationDOE OptimizationInvestigation: PDK-1Assay yield (MLR)

Response Surface Plot

MODDE 8 - 11/15/2007 1:14:36 PM

Cu/Pd ratio = 2

Cu/Pd = 2

Page 66: DOE Applications in Process Chemistry Presentation

DOE ConfirmationDOE Confirmation

Confirming reaction run using iChem Explorer to monitor reaction

DOE/Automation Improvements over HTS Result:• 70% reduction in Pd charge• 70% reduction in ligand charge• 63% reduction in Cu charge• 94% reduction in time cycle• Improved selectivity from 9:1 to 100:1-mostly due to time aspect

NH

O

I

NH

O

2

12 mol% CuI3 equiv iPr2NH

MeCN45 oC/ 1 h

99 A%

3 mol% [(allyl)PdCl]212 mol% (2-furyl)3P

+

1 A%

bis additionimpurity

Page 67: DOE Applications in Process Chemistry Presentation

iChem ExploreriChem ExplorerHardware: heating (to 150 Hardware: heating (to 150 ooC) C) and stirring block for HP and stirring block for HP 1100/1200 systems1100/1200 systems

Software: to visualize dataSoftware: to visualize data

up to 100- 1 mL reactions in LC up to 100- 1 mL reactions in LC vialsvials– monitor by direct injectionmonitor by direct injection

Page 68: DOE Applications in Process Chemistry Presentation

DOE SummaryDOE Summary

33% isolated yield

78% isolated yield

HTS/DOE (19 rxns)

NH

O

I

NH

O

2

12 mol% CuI3 equiv iPr2NH

MeCN45 oC/ 1 h

99 A%

3 mol% [(allyl)PdCl]212 mol% (2-furyl)3P

Page 69: DOE Applications in Process Chemistry Presentation

Case Study # 5Case Study # 5Cl

CO2H

OMe

Cl

CO2H

OMe

CO2H

OMe

Cl

CO2H

Cl

CO2H

CHO

+ + +

H2

Mark Weisel

10% loading Pearlman’s catalyst25 oC/45 psi/EtOAc

88 A%

Goal: to minimize formation of impurities/maximize desired product

12 A%

Page 70: DOE Applications in Process Chemistry Presentation

Case Study # 5Case Study # 5Cl

CO2H

OMe

Cl

CO2H

OMe

CO2H

OMe

Cl

CO2H

Cl

CO2H

CHO

+ + +

H2

Mark Weisel

88 A%

12 A%

DOE design: 4 Factors (19 reactions)

Temp (25 and 55 oC)Pressure (30 and 60 psi)Pd(OH)2 loading (5 and 15 wt%)Volume EtOAc (6 and 10 ml/g)

Page 71: DOE Applications in Process Chemistry Presentation

FactorsFactors

-6

-4

-2

0

2

4

6

8

10

12

14

16

18T

em

p

Pd

Vo

l

Te

mp

*Pd

Pd

*Vo

l

A%

MODDE 8 - 1/7/2008 4:30:11 PM

Page 72: DOE Applications in Process Chemistry Presentation

Effect of Pd and TempEffect of Pd and Temp

MODDE 8 - 2/1/2008 10:52:20 AM

v o l u m e = 1 0

Page 73: DOE Applications in Process Chemistry Presentation

Optimal SettingsOptimal Settings

Cl

CO2H

OMe

Cl

CO2H

OMe

CO2H

OMe

Cl

CO2H

Cl

CO2H

CHO

+ + +

H2

Pd(OH)2EtOAc

Mark Weisel

Relevant Factors-ranked

1. Pd loading ( 15 wt%)2. Temp (25 oC)3. Volume (10 ml/g)4. Pressure- no effect- run at 30 psi

Selectivity improved from 88 A% to > 99 A%

Page 74: DOE Applications in Process Chemistry Presentation

DOE BenefitsDOE Benefits

Increase your process knowledgeIncrease your process knowledge

Discover the effects of changing factorsDiscover the effects of changing factors

Understand the effects of interactionsUnderstand the effects of interactions

Learn what is and what is NOT importantLearn what is and what is NOT important

Save time, materials, and Save time, materials, and moneymoney

Page 75: DOE Applications in Process Chemistry Presentation

Take Home MessageTake Home Message

DOE is a powerful tool for optimization of DOE is a powerful tool for optimization of reactionsreactions

Automated tools minimize the effort of Automated tools minimize the effort of running multiple rxnsrunning multiple rxns

HTS & DOE in 96-well format represents HTS & DOE in 96-well format represents leading-edge scienceleading-edge science

Academics embracing HTS approachAcademics embracing HTS approach– Professor D. MacMillan (Professor D. MacMillan (Princeton)

Page 76: DOE Applications in Process Chemistry Presentation

AcknowledgmentsAcknowledgments

Peter MaligresPeter Maligres

Danny GauvreauDanny Gauvreau

Spenser DreherSpenser Dreher

Dave PollardDave Pollard

Shane KrskaShane Krska

Mark WeiselMark Weisel

Dave TellersDave Tellers

Page 77: DOE Applications in Process Chemistry Presentation

QUESTIONS ?QUESTIONS ?