Cross-Contamination in Drug Manufacturing: The Regulatory Trends
Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing...
-
Upload
carmella-dortha-merritt -
Category
Documents
-
view
214 -
download
0
Transcript of Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing...
![Page 1: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/1.jpg)
Drug Manufacturing
BIT 230
Walsh Chapter 3
![Page 2: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/2.jpg)
Drug Manufacturing
Most regulated of all manufacturing industries Highest safety and quality standards Parameters include:
– Design and layout of facility– Raw materials– Process itself– Personnel– Regulatory framework
![Page 3: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/3.jpg)
Pharmacopeias
Discussed before in other units and classes Martindale- not a standards book Gives information about drugs
– Physiochemical properties– Pharmacokinetics– Uses and modes of administration– Side effects– Appropriate doses
![Page 4: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/4.jpg)
GMP guidelines
Different publications world wide, but generally have similar information
Go over everything from raw materials to the facility
US guidelines issues publications called “Points to Consider” for additional guidelines for newer biotech products (will go over these later in semester)
![Page 5: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/5.jpg)
Manufacturing facility
Most manufacturing facilities have requirements, but some specifics to biotech products, especially
– Clean room
– Water
![Page 6: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/6.jpg)
Clean Rooms
Clean room views Environmentally controlled areas Critical steps for bio/injectable drugs are
produced in clean rooms Contain high efficiency particulate air (HEPA)
filters in the ceiling Figure 3.1 page 98 of chapter
![Page 7: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/7.jpg)
Classification of Clean Roomsfor Pharma industry
Class # microrganisms/m3 of air
A <1
B 5
C 100
D 500
See table 3.5 page 100 of chapter
![Page 8: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/8.jpg)
Other considerations
Exposed surfaces – smooth, sealed, non-penetrable surface
Chemically-resistant floors and walls Fixtures (lights, chairs, etc.) minimum and
easily cleaned Proper entry of materials and personnel into
clean room to reduce risk of contamination in clean room
![Page 9: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/9.jpg)
Gowned person in Clean room
![Page 10: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/10.jpg)
Clean Room clothing
Covers most of operators body Change in a separate room and enter clean
room via an air lock Clothing made from non-shredding material Number of people in a clean room at once
limited to only necessary personnel (helps with automated processes)
![Page 11: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/11.jpg)
CDS
Cleaning, decontamination and sanitization C- removal or organic and inorganic material
that may accumulate D-inactivation and removal of undesired
materials S- destroying and removing viable
microorganisms
![Page 12: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/12.jpg)
CDS cont’d
Done on surfaces that either are direct or indirect contact with the product
Examples of surfaces in both categories?
![Page 13: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/13.jpg)
CDS of process equipment
Of course trickier because comes in contact with the final product
Clean equipment, then rid equipment of cleaning solution
Last step involves exhaustive rinsing of equipment with pure water – WFI– Followed by autoclaving if possible– If possible use CIP (cleaning in place)
![Page 14: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/14.jpg)
Examples of CIP agents used to clean chromatography columns
0.5-2.0 M NaCl Non-ionic detergents 0.1-1.0 M NaOH Acetic Acid Ethanol EDTA Protease
![Page 15: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/15.jpg)
Water
WFI- talked about this extensively before 30,000 liters of WFI needed for 1kg of a recombinant
protein Use tap water just for non-critical tasks Purified water – not as pure as WFI, but used for
limited purposes (in cough medicines, etc.) WFI used exclusively in downstream processing Will not cover pages 105-112- water and
documentation pages
![Page 16: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/16.jpg)
Sources of Biopharmaceuticals
Genetic engineering of recombinant expression systems
Your talks will be about types of systems and how they are used- mammalian cells, yeast, bacteria etc.
Most approved products so far produced in E. coli or mammalian cell lines
![Page 17: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/17.jpg)
E. coli
Cultured in large quantities Inexpensive (relatively speaking) Generation of quantities in a short time Production facilities easy to construct
anywhere in the world Standard methods (fermentation) used
![Page 18: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/18.jpg)
Current products from E. Coli
tPA (Ekokinase) Insulin Interferon Interleukin-2 Human growth hormone Tumor necrosis factor
![Page 19: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/19.jpg)
Heterologous systems
Expression of recombinant proteins in cells where the proteins do not naturally occur
Insulin first in E. coli Remember the drawbacks of expression in
E. coli?
![Page 20: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/20.jpg)
Other problems with E. coli
Most proteins in E. coli expressed intracellularly
Therefore, recombinant proteins expressed in E. coli accumulate in the cytoplasm
Requires extra primary processing steps (e.g. cellular homogenization) and more purification (chromatography)
![Page 21: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/21.jpg)
Other problems with E. coli, cont’d
Inclusion bodies– Insoluble aggregates of partially folded product– Heterologous expressed proteins overload the
normal protein-folding machinery– Advantage- inclusion bodies are very dense, so
centrifugation can separate them from desired material
![Page 22: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/22.jpg)
Preventing inclusion bodies
Lower growth temperature (from 37C to 30C)
Use a fusion protein (thioredoxin) - native in E. coli – protein expressed at high levels and remains soluble
![Page 23: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/23.jpg)
Expression in animal cells
Major advantage- correct PT modifications Naturally glycosylated proteins produced in:
– CHO - Chinese hamster ovary– BHK - baby hamster kidney– HEK – human embryonic kidney
![Page 24: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/24.jpg)
Current products from animal cells
tPA FSH Interferon - Erythropoietin FSH Factor VIIa
![Page 25: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/25.jpg)
Disadvantages of animal cells(compared to E. coli)
Complex nutritional requirements Slower growth More susceptible to damage Increased costs
WILL NOT cover bottom of page 116 to page 124 (up to biopharmaceuticals)- you will cover these in your presentations
![Page 26: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/26.jpg)
Final Product Production
Focus on E. coli and mammalian systems Process starts with a single aliquot of the
Master Cell Bank Ends when final products is in labeled
containers ready to be shipped to the customer
![Page 27: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/27.jpg)
Production: Upstream and Downstream
Upstream: initial fermentation process; yields initial generation of product
Downstream: purification of initial product and generation of finished product, followed by sealing of final containers
biomanufacturing process overview
![Page 28: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/28.jpg)
![Page 29: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/29.jpg)
Upstream processing
Remove aliquot from MCB Inoculate sterile medium and grow (starter
culture) Starter culture used to inoculate larger scale
production culture Production culture inoculates bioreactor Bioreactors few to several thousand liters See figure 3.13 of chapter (page 129)
![Page 30: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/30.jpg)
Upstream cont’d
Pages 129-133 go over specific details for microbial fermentation
Pages 133-134 go over specific details for animal cell culture Properties of animal cells
– Anchorage dependent– Grow as a monolayer– Contact inhibited– Finite lifespan– Longer doubling times– Complex media requirements
![Page 31: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/31.jpg)
Downstream processing
Diagram page 135 of chapter 3
Detailed steps considered confidential
Clean room conditions for downstream
![Page 32: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/32.jpg)
Downstream cont’d
Steps involved (intracellular products – E. coli.) – mammalian products secreted in media, so easier to isolate)
– Centrifugation or filtration– Homogenization– Removal of cellular debris– Concentration of crude material (by precipitation or ultra
filtration)– High resolution chromatography (HPLC)– Formulation into the final product
![Page 33: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/33.jpg)
Downstream cont’d
Final product formulation– Chromatography yields 98-99% pure product– Add excipients (non active ingredients), which
may stabilize the final product– Filtration of final product, to generate sterile
product– Freeze drying (lyophilization) if product if to be
sold as a powder (dictated by product stability)
![Page 34: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/34.jpg)
Separation methods
Page 142,tables 3.18 and 3.19 Familiar with:
– Ion-exchange– Gel-filtration– Affinity chromatography
Protein A chromatography Immunoaffinity chromatography
![Page 35: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/35.jpg)
Factors that influence biological activity
Denature or modify proteins Results in loss of/reduced protein activity Need to minimize loss in downstream work Problems can be chemical (e.g., oxidizing,
detergents); physical (e.g., pH, temperature); or biological (e.g., proteolytic degradation)
Table 3.20 page 143
![Page 36: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/36.jpg)
Proteolytic degradation
Hydrolysis of one or more peptide bonds Results in loss of biological activity Trace quantities of proteolytic enzymes or chemical
influences Several classes of proteases:
– Serine– Cysteine– Aspartic– Metalloproteases (also in other ppt)
![Page 37: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/37.jpg)
Protease inhibitors
PMSF – serine and cysteine proteases Benzamidine – serine proteases Pepstatin A – aspartic proteases EDTA – metalloproteases
a.a residue known to be present at active site of protein, so disruption of it causes loss of activity
![Page 38: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/38.jpg)
Others (mentioned before)
Deamidation – hydrolysis of side chain of asparagine and glutamine– Happens at high temp and extreme pH
Oxidation and disulphide exchange– Oxidation by air (met and cys in particular)
Alterations of glycosylation patterns in glycoproteins (more than one sugar)– Affect activity or immunological properties
![Page 39: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/39.jpg)
Excipients
Substances added to final product to stabilize it
Serum albumin– Withstands low pH or elevated temps– Keeps final product from sticking to walls of
container– Stabilize native conformation of protein
![Page 40: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/40.jpg)
Excipients cont’d
Amino acids
– Glycine – stabilizes interferon, factor VIII, stabilizes against heat
Alcohols (and other polyols)– Stabilize proteins in solution
Surfactants– Reduces surface tension; proteins don’t aggregate, so don’t denature
![Page 41: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/41.jpg)
Final product fill
See figure 3.27 page 153 Bulk product gets QC testing Passage through 0.22 m filter for final
sterility Aceptically filled into final product containers Uses automated liquid handling systems
![Page 42: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/42.jpg)
Final product fill cont’d
Freeze drying (lyophilization) Yields a powdered product Reduces chemical and biological degradation
of final product Longer shelf life than products in solution Storage for parenteral products (those
administered intravenously or injected)
![Page 43: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/43.jpg)
Freeze drying cont’d
Need to add cryoprotectors– Glucose or sucrose– Serum albumin– Amino acids– Polyols
Freeze drying can be done in many steps
![Page 44: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/44.jpg)
Labeling and Packing
After sealed in final container, product quarantined
Samples are QC’d Check potency, sterility and final volume Detection and quantitation of excipients Highly automated procedures Labeling function critical- biggest error where
many products are made
![Page 45: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/45.jpg)
Label
Name and strength of product Specific batch number Date of manufacture and expiry date Required storage conditions Name of manufacturer Excipients included Correct mode of usage
![Page 46: Drug Manufacturing BIT 230 Walsh Chapter 3. Drug Manufacturing Most regulated of all manufacturing industries Highest safety and quality standards Parameters.](https://reader030.fdocuments.in/reader030/viewer/2022032803/56649e215503460f94b0d184/html5/thumbnails/46.jpg)
Other final product items
Biopharmaceutical products undergo more testing than traditional pharma products
Products made in recombinant systems have more potential to be contaminated than synthetic chemical drugs
Larger, more complex molecules