Biocatalysis: A valuable tool
for cost savings in the synthesis of pharma intermediates and APIs
Oliver May Competence Manager Biocatalysis San Francisco, January 26th
Content
Technology:
- Introduction to Biocatalysis and Biodiversity
Application:
- Divers and Challenges - Examples
Summary:
- Two take home messages
Some Statements about Biocatalysis
The overall impact [of biocatalysis] on the chemical process industry will be slight. Organic chemistry will continue as its mainstream.
R. L. Hinman, A Role for Biotech in Producing Chemical? Bio/Technology 1991, 9, 533 (1991)
Only 10 years later
biocatalysis, we are no longer limited In by our ignorance. We are only limited by our imagination.
Frances H. Arnold Caltech, 1999
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Biocatalysis is emerging
Tremendous progress in e.g.
<1990
black box era
What happened?
>2000
knowledge based era
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Structural Biology recDNA technology Automation DNA sequencing Genome projects Bioinformatics Metagenomics Synthetic genes Synthetic diversity
Biocatalysis is emerging
Structural Biology recDNA technology Automation DNA sequencing Genome projects Bioinformatics Metagenomics Synthetic genes Synthetic diversity
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Biocatalysis is emerging
Structural Biology recDNA technology Automation DNA sequencing Genome projects Bioinformatics Metagenomics Synthetic genes Synthetic diversity
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Biocatalysis is emerging
Structural Biology recDNA technology Automation DNA sequencing Genome projects Bioinformatics Metagenomics Synthetic genes Synthetic diversity
Several 200 l ocean water from Sargasso Sea: over 1.2 million previously unknown genes discovered
Venter et al. Science (2004): Vol. 304. no. 5667, pp. 66 - 74
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Our Biocatalysis Toolbox is rapidly growing
Past Present Future
Trends in Biocatalysis
The number of applications will be growing due to significant advances in technology (e.g. access to biodiversity, enzyme engineering, )
Biocatalysis fastest growing
s on ti ic ed Pr
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About Predictions
forecasting the future of scientific research In there is one quite general law to be noted. The unpredicted always happens. So one can be quite sure that the future will make any detailed prediction look rather silly.
J.B.S. Haldane (1892-1964) in The Future of Biology
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About Predictions
believe in horses. I The automobile is a transient phenomenon.
Wilhelm II, last German emperor (1905)
First invented car (Carl Benz, 1886)
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Trends in Biocatalysis
Growth of Enzymatic Processes 3000
The number of applications will be growing due to significant advances in technology (e.g. access to biodiversity, enzyme engineering, )
# of Patent Families
2500 2000 1500 1000 500 0 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 Year
Outside DSM
Biocatalysis fastest growing
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Growth of alcohol dehydrogenase targets
# of requests for ADHs
9 8 7 6 5 4 3 2 1 0
s on ti ic ed Pr
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lity ea R
Q3 Q4 Q1 Q2 2006
Inside DSM
Q1 Q2 2005
Q3
Q4
Q1 2007
From DSM track record to our believe in Bio
>100 products and applications
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DSM Biocatalysis Competence in Depth & Breath unmatched!
Combined strengths and experience makes DSM unique in biocatalysis
Strain devel. Enzyme devel. Fermentation. Enzyme manufacturing
Delft, Kaiseraugst
Capua
Chemistry Route scouting Platform screening
Geleen
Linz, Venlo, Regensburg
Enzyme application Regulatory (cGMP)
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Outside world: Which Companies will be successful in Industrial Biotechnology in the Long Run?
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Content
Technology:
- Introduction to Biocatalysis and Biodiversity
Application:
- Divers and Challenges - Examples
Summary:
- Two take home messages
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Driver: Cut down steps Improve efficiency
Opportunity: Cut down steps by clever chemistry!
Source: Org.Biomol.Chem., 2006, 4, 2337-2347 17
Pharma Need:
efficient manufacturing processes for very complex products
Pharma Syntheses are Multi-step Syntheses!
Opportunity: Cut down steps by clever chemistry!
Source: Org.Biomol.Chem., 2006, 4, 2337-2347
Asymmetric Synthesis
(e.g. HCN addition, ketone reduction, transamination, NH3 addition)
100% yield >99.9% ee
Desymmetrization
(e.g. nitrile,18 ester, epoxide hydrolysis)
Dynamic kinetic resolution & deracemization
(e.g. hydrolysis of hydantoins, esters or acylation of alcohols, redox cycles for amines and alcohols)
Examples of established pharma processes based on enzymatic transformations
Product Reaction Type reduction; Enzyme Platform Dorzolamide; Nelfinavir; Pregabalin;
desymmetrization;
stereosel. hydrolysis;
Cholest. lowering; C-C bond formation;
Cardiovascular; asymm. cyanide addition;
10 0%
Pyrethroide;
stereosel. hydrolysis;
2n dG en e
Antibiotic; Blood Pressure;
dynamic kinetic resolution; sterosel. hydrolysis;
Ma ny
Cardiovascular; asymmetric NH3 addition;
Substitution due to significant cost savings! First time right is still exception 19
dif Incr fer ea en sin tp gn lat um fo rm ber s a of pp lie d
ketoreductase epoxide hydrolase lipase or nitrilase aldolase HNL amidase ammonia lyase hydantoinase
ra tio np ro ce ss es
co nc ep ts
hydrolase (esterase) DSM Examples
Typical Development Timelines
Substrate / product preparation
ca. 2-4 weeks
Develop analytics (%ee, yield)
ca. 2 weeks
Identification of enzyme which - Accepts substrate - Shows desired selectivity
Enzyme screening
ca. 2-4 weeks
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Typical Development Timelines
Substrate / product preparation
ca. 2-4 weeks
to nt Develop analytics re y ! (%ee, yield) iffe -lBiocatalyst preparation d o og ot hn - Reaction engineering n c ca. 2 weeks - Process development es r t e n Enzyme - Sample supply eli screening he - Large-scale quotation im o t T
Go / no go for
ca. 2-4 weeks
ny a
ca. 1,5-2,5 months
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ca. 2-3 months
Success factors
l
Broadest collection of off-the-shelf enzymes
700 600 Number of enzymes 500 DSM plus partners 400 300 200 100 0 1 2 3 4 es ses ses es s cta yas fera ydrola l u s h red ran t o 22
supplier 1 Biocatalytics
Sigma-Aldrich supplier 2
supplier Codexis
oxi
Success factors
l
Broadest collection of off-the-shelf enzymes established and expanded @ DSM through:
In-house development (Corporate Research) Partnering (Verenium, IEP, Libragen,...) Use of suppliers (Codexis, Novozymes, Amano, ) Academic network (Graz, Manchester, IBOS, B-BASIC )
Increase chance to identify suited enzyme Reduce development cost Timelines fit to pharma needs
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Success factors
l l
Broadest collection of off-the-shelf enzymes Secure supply at any scale (and with FTO!)
In-house enzyme production (Capua >1.400.000 l)
Broad range of proprietary Production organisms (pluGbugs)
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Success factors
l l l
Broadest collection of off-the-shelf enzymes Secure supply at any scale (and with FTO!), fast scale-up Experience (also regulatory) in large-scale implementation
Biocatalysis Platforms
L -Amidases D -Hydantoinase / D-C arbamoylase R - and S -hydroxynitrile lyases
Product Classes
-Amino acids, -Hydroxy acids -Amino acids -/-Amino acids, -Hydroxy acids
ed ent -Amino acids m Acylases -/-Amino acids, Amines, Amino-nitriles ple im Lipases, Esterases Carboxylic acids, Alcohols, Amines es Proteases Peptides ss Aldolases 1,2-Diols, Aminoalcohols, Hydroxyesters oce r Epoxide hydrolases al p Epoxides, 1,2-Diols tri Racemases, Isomerases us Racemization Haloperoxidases Alcohols, Epoxides, Sulfoxides ind 5 Hydratases >2 -/ -Hydroxy acids
Ammonia lyases Nitrilases Reductases & Dehydrogenases -/ -Amino acids, -Hydroxy acids Alcohols, other chiral intermediates
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Success factors
l l l l
Broadest collection of off-the-shelf enzymes Secure supply at any scale (and with FTO!), fast scale-up Experience (also regulatory) in large-scale implementation Interdisciplinary route scouting (including all options!)
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DSM: Leader in Industrial Biocatalysis
Examples
Biocatalysis Platforms
L -Amidases D -Hydantoinase / D-C arbamoylase R - and S -hydroxynitrile lyases
Product Classes
-Amino acids, -Hydroxy acids -Amino acids
Ammonia lyases Acylases Lipases, Esterases Proteases Aldolases Epoxide hydrolases Racemases, Isomerases Haloperoxidases Hydratases
>2
ind 5
ted en -Amino acids lem -/-Amino acids, Amines, Amino-nitriles impAlcohols, Amines Carboxylic acids, ses Peptides es Aminoalcohols, Hydroxyesters roc p 1,2-Diols, 1,2-Diols l Epoxides, ria t Racemization us
-/-Amino acids, -Hydroxy acids Alcohols, Epoxides, Sulfoxides -/ -Hydroxy acids -/ -Amino acids, -Hydroxy acids Alcohols, other chiral intermediates
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Nitrilases Reductases & Dehydrogenases
Example for Dynamic Kinetic Resolution
Hydantoinase Technology
Now also S!
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Example for Enzymatic Desymmetrization
O R 1O O
pig liver esterase (PLE) O
OR 1 HO R2
O OR1
R2
R3
R3
DSM has exclusive access to a non-animal derived porcine liver esterase!
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DSM Innovation Award 2007
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Example for Asymmetric Synthesis (1)
Ammonia Lyase Technology
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Example for Asymmetric Synthesis (2)
Hydroxynitrile Lyase Technology
Source Almonds Rubber Tree
StereoSpecificity R S
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Substrate Specificity Aliphatic, aromatic Aliphatic, aromatic
Platform offers Access to a wide Spectrum of Intermediates
O R1 R2 Hydroxy Nitrile Lyase NC O H R1 R2
H+ O H 2N R1 OH R2 Nitrilase Nitrile Hydratase NC O H R1 R2 H+ [H]
HOOC OH R1 R2
[H]
NH 2 OH R1 R2 OH OH R1 R2 O R1 R2
HOOC OH R1 R2
[H]
NaNO 2
>100 unique nitrilases in our screening platform
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Example for Asymmetric Synthesis (3)
Aldolase Technology
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Example for Asymmetric Synthesis (4)
Dehydrogenase Technology
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Example for Asymmetric Synthesis (5)
Enone Reductase Technology
R2 R1 R3
NAD(P)H NAD(P) H+ [H-]
O R4 R1
R2
O R4 R3
R2 R1 R3
OR4 R1
R2
O R4 R3
E/Z (R/S)
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broad
Content
Technology:
- Introduction to Biocatalysis and Biodiversity
Application:
- Divers and Challenges - Examples
Summary:
- Two take home messages
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Conclusions
Toolbox of Biocatalysis is rapidly developing - More versatile due to better access to biodiversity - Proven and well established within DSM Don be afraid to kiss the t Biocatalysis Frog - It can be fast - It can be highly rewarding - If you are still afraid to kiss ask us ,
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For further information about biocatalysis, please contact: Name Phone E-mail Dr. Oliver May +31 46 47 60551 [email protected]
