Purification and Isolation Technologies - Hovione

Purification and Isolation Technologies Ricardo Mendonça & Rui Loureiro 27.Jan.2016

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Purification and Isolation Technologies

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Confidential Hovione © 2016

Rui Loureiro Rui Loureiro joined Hovione in 2008 as a Process Chemist and

after several positions he is currently the Director of the R&D Process Chemistry area where is responsible for the development and scaleup of processes to produce active pharmaceutical ingredients under development. Rui Loureiro has a degree in Physics and Chemistry from the University of Algarve, Portugal and a Ph.D. in organic chemistry from the University of Liverpool, UK. After completing his Ph. D. Rui continued in the University of Liverpool as a post-doc fellow before joining Lintfield Ltd as a process chemist where he stayed between 2004 and 2008.

In his work Rui Loureiro as published ten papers in peer review magazines and is author of four patents. He has also carried out several oral (8) and poster communications (10). Currently his interests are scaling-up of processes to produce and purify APIs under a quality by design approach, flow chemistry and process modelling.


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Ricardo Mendonça Ricardo Mendonça joined Hovione in 2007 as a process chemist and is currently a Group Leader in the R&D Process Chemistry Development area where is responsible for the development and scale-up of processes to produce Active Pharmaceutical Ingredients (APIs) under development.

Ricardo has a degree in Biochemistry at the University of Algarve in 1998. Afterwards he moved to Liverpool UK to carry out a Ph.D. in Organic Chemistry. After completing his Ph.D. Ricardo moved to Tempe, Arizona as a post-doc fellow working at the Cancer Research Institute, under the supervision of Prof. George R. Pettit, in the area of Natural Products Synthesis. After moving to Portugal Ricardo worked between 2005 and 2007 at the New University of Lisbon working in the development of new asymmetric brominating reagents.

Ricardo Mendonça has published nine scientific papers and is the author of one patent. He has also carried out several oral and poster communications. Ricardo also contributes to the scientific community as a reviewer of scientific papers and as an invited lecturer of Industrial Synthesis of APIs at the Faculty of Pharmacy (University of Lisbon). Currently his interests are biocatalysis, carbohydrate chemistry, peptide synthesis and large-scale chromatography. Purification and Isolation Technologies

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Agenda

Introduction Purification Technologies Purification by IEX – Case Study Isolation technologies Integrated Purification & Isolation QbD applied to purification and isolation


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Introduction – Complexity increasing

New chemical entities are becoming more complex and more difficult to purify and isolate Bryostatin 1

Telavancin

Dolastatin 10

Enfuvirtide Purification and Isolation Technologies

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Introduction – Complexity increasing Enantiomeric separation

Purification of complex mixtures

Long synthesis

Purification and Isolation Challenges

Particle Design

Removal of specific impurities such as GTI’s


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Agenda



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Introduction – Purification: Crystallization Crystallization has been at the forefront of purification technologies for a long time: • From the melt o Not widely used • From solution o Temperature controlled o Evaporation controlled o Anti solvent addition

New trends: •

Sonocrystallization o The Use of Ultrasound for Improved Industrial • Crystallization o The most important effect of ultrasound on crystallization is the induction of nucleation and the principal benefits derived from an ability to manipulate this effect.


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Continuous Flow o Emerging technique o Offers several benefits • Cost reduction • Faster scale-up • Controllable quality Confidential Hovione © 2016

Introduction – Purification: Chromatography Chromatography is a common technology used industrially to purify a whole lot of compounds. In the pharmaceutical arena it’s mostly used industrially to purify complex compounds and pure enantiomers Several (LC) types: Low-pressure (up to 3 bar), Medium pressure (up to 240 bar),

High-pressure By mode of separation:

• • • • •

Specific binding interactions (affinity chromatography) Charge (ion exchange chromatography) Size (size exclusion chromatography/gel filtration chromatography) Hydrophobic surface area (hydrophobic interaction chromatography and reverse phase chromatography) Multiple properties (multimodal or mixed-mode chromatography) Purification and Isolation Technologies

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Introduction – Purification: New Trends in Chromatography

Simulated moving bed (SMB) •

Multiple smaller columns containing the solid adsorbent (beds) rather than one large column, and to “move” the beds in the opposite direction of the fluid to achieve a countercurrent flow, rather than flowing fluids through one static bed. Allows much higher productivity (20x) relative to batch (single-column) methods.

Drugs purified using SMB: • Prozac, • Citaprolam

Centrifugal Partition Chromatography (CPC) • •

Liquid-liquid chromatography technique based on partition of compounds between two immiscible liquide phases. Has been used for the purification of oligonucleotides (WO2013030263 A1)

Supercritical fluid chromatography (SFC)


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Introduction – Purification: Membrane separation processes

Extremely useful technique for the removal of water from solutions of APIs. Can be used to remove dissolved gases, salts, impurities etc. Depending on the size of compounds present and their difference several types of membranes are available for: • • • • •

Reverse osmosis Nanofiltration Ultra filtration Microfiltration Particle filtration Purification and Isolation Technologies

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Introduction – Purification: New Trends in Membrane separation processes

Organic solvent nanofiltration (OSN) is emerging as a novel method to remove organic impurities from APIs. Use of molecularly imprinted polymers (MIPs) and synergistic combinations of these two technologies is also a hot research field.

These technologies allow reduced energy consumption, improved process capability, low maintenance, increased automation, modularity and reduced footprint in the purification of APIs.

Green Chem., 2014, 16, 4440–4473


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Introduction – Purification: New Trends in Membrane separation processes

NEMOPUR (New Molecular Purification Technology for Pharmaceutical Production)

European Commission FP7funded Marie Curie Initial Training Network of nine partner organisations from industry and academia

Organic solvent nanofilotration: A platform for removal gentoxins from active pharmaceutical ingredients; Journal of Membrane Science 381 (2011), 21-33 A hybrid approach to reach stringent low genotoxic impurity contents in active pharmaceutical ingredients: Combining molecularly imprinted polymers and organic solvent nanofiltration for removal of 1,3diisopropylurea; Separation and Purification Technology 86 (2012), 78-87 Design, preparation and characterization of novel molecularly imprinted polymers for removal of potential genotoxic 1,3-diisopropylurea from API solutions; Separation and Purification Technology 86 (2012), 190-198 Purification and Isolation Technologies

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Agenda



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Purification – IEX case study Scaling-up

Chromatographic processes:  Efficient product purification  Resins can be re-used  Different types of resins can be used Elution monitored by PAT tools; QbD approach

Packing may be challenging; Uses large volumes of solvent;

Crude API: ~85-90% purity

Final API: ~99.5% purity


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Purification – IEX case study

•

Chromatography scaling-up challenges: • Column size • Stationary phase used • Wall effect • Pressure

• Mobile phase flow • Gradient • Elution rate variation from small scale to large scale • Column packing – ensuring it is properly packed


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Purification – IEX case study Laboratory scale •

Starting point ! •

Preparative chromatography system

•

Binary gradient pumping system (2.5 to 250 ml/min)

•

Pressure max 50 bar

•

Fraction collector

•

UV detector (200-840 nm) with 4 simultaneous channels

•

Controlled via SepacoreControl software


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Purification – IEX case study Resin selection Ion exchange resins are highly ionic, covalently cross-linked, insoluble polymers supplied as beads. The beads have either a dense internal structure with no discrete pores (gel resins, also called microporous resins) or a porous, multichannelled structure (macroporous or macroreticular resins).


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Purification – IEX case study Resin selection Ion exchange resins are highly ionic, covalently cross-linked, insoluble polymers supplied as beads. The beads have either a dense internal structure with no discrete pores (gel resins, also called microporous resins) or a porous, multichannelled structure (macroporous or macroreticular resins). • Several types of resins available: o

Strongly acidic (i.e. sulfonic acid groups)

o

Weakly acidic (i.e. carboxylic acid groups)

o

Strongly basic (i.e. quaternary amino groups)

o

Weakly basic (i.e. primary, secondary and ternary amino groups)

o

Ampholytic containing acidic and basic groups

o

Selective and chelating resins Purification and Isolation Technologies

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Purification – IEX case study Scale challenges Scale available: Each scale presents its own challenges!


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Purification – IEX case study Filling properties Wall effect


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Purification – IEX case study Resin properties Pressure Flow curves

Column diameter 60 cm 4.00

Column diameter 10 cm

4.0

3.50 3.5

3.00 Pressure (bar)

3.0

Pressure (bar)

2.5 2.0 1.5

2.50 2.00

1.50 1.00

1.0

0.50

0.5

0.00 0

10

20

30

0.0 0

100 Empty Column

200 Flow (cm/h) Packed Column

300

50

60

70

Flow (cm/h)

400 Bed only

40

Empty column

Packed Column

Bed Only

At smale scale the pressure in the column increases in a linear manner

•

At large scale the pressure in the column increases in a linear fashion to a point and then it is exponential, indicative that there is no wall effect at this scale. Purification and Isolation Technologies

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80

Purification – IEX case study Resin properties

Critical velocity

Pressure Flow curves



4.0

3.50 3.5

3.00 Pressure (bar)

3.0

Pressure (bar)

2.5 2.0 1.5

2.50 2.00

1.50 1.00

1.0

0.50

0.5

0.00 0

10

20

30

0.0 0

100 Empty Column


300

50

60

70

Flow (cm/h)

400 Bed only

40

Empty column

Packed Column

Bed Only


•


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Max packing flow




4.0

3.50 3.5

3.00 Pressure (bar)

3.0

Pressure (bar)

2.5 2.0 1.5

2.50 2.00

1.50 1.00

1.0

0.50

0.5

0.00 0

10

20

30

0.0 0

100 Empty Column


300

50

60

70

Flow (cm/h)

400 Bed only

40

Empty column

Packed Column

Bed Only


•


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80


Elution flow




4.0

3.50 3.5

3.00 Pressure (bar)

3.0

Pressure (bar)

2.5 2.0 1.5

2.50 2.00

1.50 1.00

1.0

0.50

0.5

0.00 0

10

20

30

0.0 0

100 Empty Column


300

50

60

70

Flow (cm/h)

400 Bed only

40

Empty column

Packed Column

Bed Only


•


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Purification – IEX case study Packing Column packing Is one of the most important aspects that should be taken in account when scaling up a chromatographic process Irregularities may cause:

•

Uneven flow within the bed

•

Band broadening (resolution lost)

•

Zone mixing

•

Changes in flow rates


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Purification – IEX case study Packing How to check column packing? • Visual inspection (i.e. dye) • Determining plate numbers/HETP, Asymmetry (Efficiency)


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Purification – IEX case study Packing Pulse experiments

Replicate #1

Replicate #2

60000

54000 52000

55000

using NaCl

50000 48000

50000

46000 45000

44000 42000

40000 35000 00:00:00

40000 00:14:24

00:28:48

Results Vmaximum W half height Assymetry factor Number plates HETP RPH

0.5973 0.04629 0.58 922 0.06 3.05


00:43:12

00:57:36

38000 00:00:00 00:14:24 00:28:48 00:43:12 00:57:36 01:12:00 01:26:24

01:12:00

Recommended values

0.8-1.8