Byrn Lectures Flashcards
Polio Vaccine
- brought to public in 1955
- bore a Lilly label
- Polio cases dropped from 1000’s to 53
4 Types of COVID Vaccines
mRNA
Vector
Protein Subunit
Whole, killed
mRNA
- used by Pfizer, Moderna
- mRNA used to instruct cells to make a piece of spike protein
- coated in fatty lipids to protect it
Vector
- used by Janssen, AstraZeneca, Sputnik
- used adenoviruses to carry genetic instructions for spike protein
- attach to cells and inject DNA to tell cells to make spike protein
Protein Subunit
- used by Novavax, Sanofi
- get small pieces of the target virus circulating into the system by engineering insect viruses used to infect moths that the cells are used then to create spike protein
Whole, Killed
- used by Sinovac
- batches of the virus are grown and then killed using chemicals or heat
Novavax COVID/FLU combo Vaccine
- launched a phase 1/2 trial
- tested in 640 healthy adults aged 50-70
- must have been injected by COVID-19 or vaccinated
Stability of RNA
-70 degrees is stable at this temperature over 32 days
RT is degraded over 32 days (can be thawed for an hour and be given at RT safely without degradation)
Ionizable Lipids
- capsulate RNA in lipids to prevent being hydrolyzed
- positive charged at acidic at pH
- neutral in the blood
Ionizable Cationic Lipid
ALC-0315 (Pfizer) & SM-102 (Moderna) at a pH of 6.2-6.4
Helper Lipids
DSPC, Cholesterol, Zwitterionic at pH of 7.5
PEG-Lipid
ALC-0159 (Pfizer) & PEG-DMG (Moderna)
Manufacture
- dissolve lipids in ethanol in solution 1
- dissolve nucleic acid (RNA) in buffer in solution 2
- mix the two solutions together
- FORMS LIPID NANOPARTICLES (dense)
Moderna Patent
- Luciferase: protein in lightening bugs
- recognition tactic to show if the mRNA was formed
- cationic lipid + DSPC + cholesterol + PEG-lipid
- diluted to 0.2 mg/ml in a citrate buffer and then the ethanol was mixed with RNA.
- Ethanol was then removed and replaced with PBS by dialysis
- Ratio
50:10:38:1.5 - Total Lipid:RNA ratio
10:1
Microfluidics
- inject 3 green tubes injecting drug into plate making the nanoparticle
Solute
material being dissolved
typically a solid
Solvent
used for drugs to dissolve the solid
ex. alcohol, water, buffers, isopropyl alcohol, glycerine, propylene glycol
Oral Solutions
drug in solvent including water
The solution dosage form reaches higher blood levels than the solid dosag form because:
The drug is already in solution
It takes longer for solid to dissolve
Syrup
- a drug in solution containing sugar or a sugar substitute
- may contain antimicrobial preservative
Elixir
clear sweetened solutions containing a drug and alcohol
Nonaqueous Solutions
Liniment
Biological Products
Peptides - insulin
Monoclonal Antibodies - Humira
Proteins
Immune enhancing agents
Cell Therapy
Vaccines
Biological Formulations
typically solutions or freeze dried (lyphilized)
Insulin Ingredients
Glycerine, Metacresol Phenol –> Stabilizers
Zinc –> Complexing Agent
Protamine Sulfate –> Delay of Absorption
Sodium Phosphate –> Buffer
HCl, NaOH –> pH Adjusters
Common Biological Dosage Forms
- Solutions in vials
- Freeze dried vials with power (must add water)
Stress of Freezing
- feared quality of issues are aggregates
- aggregates can cause immune responses
- higher OD, more aggregates
- mediate aggregates by different pH levels during cooling
Surfactants
Polysorbate 80 allows us to stabilize the aggregates
Small Molecules vs Antibodies
Small
- structure is simple and well-defined
- predictable chemical process
- identical copy can be made
- easy to characterize
- stable
- low potential of immunogenicity
Antibody
- structure is complex, with many PTM
- each manufacture is unique
- similar copy can be made but not identical
- difficult to characterize due to mixture of related molecules
- sensitive to storage and handling
- high potential of immunogenicity
Sensitivity of mAbs
- PTM are affected by manufacturing processes
- PTM can effect mechanism of action, bioavailability, clearance, immunogenicity, effector functions, and binding
Drug Product Manufacturing
- sterile filtration of drug to get out the microbes
- container closure integrity and possibility of lyophilized
Container Closure Considerations
- depyrogenation of glass vials (dry heat and autoclaving)
- Steam and pressure sterilization
- Test materials
- Leaching and hemolysis
Drug Substance Chacterization
- should be positive for identity and specified for purity, potency, and microbial contamination
- acceptance criteria should be established
- results of stability should be given to IND
- Data of characterization should be given to IND
Specifications of Drug Substance
Identity
Purity
Impurities
Potency
Quantity
Safety
Specifications for Drug Product
Appearance
Identity
Purity
Quantity
Potency
Safety
Physical
Particulate
BioProcess Design Criteria
Quality: purity, sequency, glycosylation, activity
Concentration
Productivity
Yield
Advantages and Disadvantages of Cell Lines
Advantage
- produce human-like proteins
- secrete proteins
- correctly constructed and active proteins
Disadvantage
- slow growth rate
- low cell densities and productivity
- sensitive to fermentation conditions
Type of Cell Lines Effect
Feeding Regime
Down-Stream Processing
Scale
Bioreactor
Type of Bioreactor Depends On…
Are cells anchored or suspended?
Cell Density Mixing –> homogenous conditions throughout
Oxygen transfer rate
CIP/SIP
Shear Resistance
Process of Fermentation
Definition of Product
Selection of Producing Organism
Strain Screening
Medium Optimization
Small Scale Bioreactor
Control Requirements
Scale-Up
Types of Chromatography
Streamline
Index
Chromaflow
BPG
FineLINE
BioProcess Stainless Steel
SEE SLIDE 49
ICH Documents for Biologics
Q5 A: viral safety
Q5 B: genetic stability
Q5 C: product stability
Q5 D: cell substrate
Q5 E: comparability
Q6 B: specification
M4/M2: CTD
Q7: GMP for API
Q8: Pharmaceutical Development
Q9: Quality Risk Management
Q10: Pharmaceutical Quality System
Q11: Development and Manufacture of Drug Substance
