DD Exam 4 PDF

I liked the color coding of the last one, please feel free to use your color :) Yes… they are all custom. How whimsy! HI mitch! Hey Jared! I feel left out… - Inez NOT SO WHIMSY NOW IS IT? Bethany, Mitch, Evan, Jared, Inez Objectives Discuss the anatomy and physiology of the pulmonary system in...

I liked the color coding of the last one, please feel free to use your color :) Yes… they are all custom. How whimsy! HI mitch! Hey Jared! I feel left out… - Inez NOT SO WHIMSY NOW IS IT? Bethany, Mitch, Evan, Jared, Inez Objectives Discuss the anatomy and physiology of the pulmonary system in relationship to the aerosol particle size and deposition in the lungs ○ Main determinant of deposition is particle size ○ > 10 um for nasopharyngeal area ○ 3 um for normal deposition in lower lungs ○ We lose most of the med in the nose/mouth, so only a small percent actually makes it all the way to the bottom of the lungs Describe the differences between the types of aerosol devices and explain what the advantages and disadvantages are of each aerosol devices ○ Ultrasonic Nebulizer → piezoelectric principle, the nebulizer produces heat so it can breakdown drug/proteins, not good for microemulsions Advantages → small size (portable), short treatment time, smaller drug amount but no diluent Disadvantages → expensive, fragile, needs electricity ○ SVN → gas powered, evaporation + recirculation = stronger [drug], has a dead volume you must combate w diluent (won’t aerosolized below certain amount) → can be used in children < 2 Advantages → can aerosolized many things/mixtures, good for young/old/debilitated bc low coordination/low inspiration rate/no need to hold breath Disadvantages → expensive, cumbersome, long treatment time, contamination if not cleaned and can spray if mask doesn’t fit ○ MDI → pressurized canister of drug with suspension, co-solvent, and micronized powder, surfactants and HFA, use in people > 5  Two phase system: pressurized gas (think whipping cream) the amount of spray force decreases over time (this would decrease the amount of drug delivered)) Single phase system: constant drug delivery over time Advantages → portable/small, short treatment time, easy to use Disadvantages → hand-breathing coordination required, fixed [drug], depletion is hard to measure, can have rxns to excipients Factors impacting performance Shaking canister to redistribute creaming/suspension Time between breaths to allow for bronchodilation (5-10 min) Open vs closed mouth → open = slower particle movement and less oropharyngeal impact Loss of prime/degradation over time Temp degradation (keep between -20 to +20 degrees celsius) ○ MDI reservoir → allows for simplified coordination, slowing of particles/smaller particles, allows time for vaporization of propellant, use for people > 4, < 4 need reservoir and mask, and neonates need endotracheal tube These are usually a chamber (spacer) added to the mouth part of the inhaler Advantages → reduced oropharyngeal drug loss, use during airflow obstruction, masks for kids Disadvantages → cumbersome, added expense, assembly required and possible contamination ○ Combivent Respimat → no propellants (mechanics with spring), not for use in people with allergies to soy/peanuts as these are sometimes used ○ DPI → drug in powder form, need a high inspiration rate, not for children < 4 Turbohaler = sensitive to humidity Diskus = has powder in blister strips so more resistant to moisture Advantages → small/portable, short treatment time, no coordination or CFC, easy to determine remaining doses What does this stand for?? Chlorofluorocarbons → they were used as propellants  Disadvantages → limited drug range, need to be able to strongly inhale, high oropharyngeal deposition Have knowledge of: ○ Structure of proteins and peptides Amino acids are the building blocks of proteins and are made of an Amino, Hydrogen, Carboxyl, and R group Protein structure = function Primary → Ochem “line and dot” kind of thing Secondary → DNA Tertiary → 3D of a single peptide chain Quaternary → 3D of multiple chains together ○ Stability of proteins and peptides Easily degraded via temp changes either way, Physical instability Denaturation → change in 2 or 3 structure where stabilizing forces are impeded by temp, pH, organic solvents Surface adsorption → interaction with glass/plastic/rubber, where the molecules of peptide/drug stick to the surface of their container and ultimate loss of peptide ○ Caused by pH, container, charges, prolonged exposure but after a monolayer forms we lose less drug after that ○ Reversibility dependent on time, temp, agitation ○ Determined by SA ○ Insulin adsorbed onto glass or plastic IV tubing ○ Lessen by adding surfactants (albumin, Tween/Span, ethylene oxide) or Teflon (polyethylene oxide) Protein aggregation → often end in denaturation of product or precipitation on macroscale, caused by shaking/adsorption Chemical instability  Deamination → degradation of asparagine and glutamine in water, decrease by putting in pH = 6 and develop a lyophilized formula where we freeze dry Oxidation → can happen to many amino acids like Meth, Cyst, His, Tryp, Tyr, making them “D” can deactivate ○ Minimize by keeping from light, put in fridge, or add antioxidants True antioxidant → phenolic compounds like BHT, BHA, propyl gallate Second class antioxidant → reducing agents that become ox instead of drug like methionine, vitamin C, Na sulfite, thioglycerol ○ Pharmacokinetics of biologicals Clearance is enzyme catalyzed and have short half life with high metabolism and elimination with catalytic activity dependent on pH, temp, and [substrate/enzyme] Negative charge and increasing size decreases kidney filtration rate ○ Routes of administration of proteins and peptides Oral → not for use for protein due to stomach acid, to increase bioavailability add protease inhibitor, drug encapsulation, prodrug, absorption enhancers Parenteral → best way to admin protein and bypass biological barriers, however it is uncomfortable, can cause injuries and rxns Do not freeze pitocin or procrit Nasal and pulmonary → good place for absorption due to blood vessels and proximity to brain and increased bioavailability, avoids first pass metabolism, peptides have < 5% absorption, can cause irritation ○ Handling of protein and peptide drugs Sterility must be maintained when no preservative are used and must be pyrogen free  Be careful with filters as they can catch product - consider adding a drug distal to filter or filters made of cellulose, difluoride, polycarbonate ○ Regulatory aspects of biologicals Store in fridge (2-8 degrees), freezer (-20 degrees), room temp (15-25 degrees), ultra freezer (-80 degrees) Have very short half life Water containing compounds BUD = 14 days in fridge The advantages and limitations of suppositories ○ Advantages → painless/easy to insert, first pass partially avoided (the further up you push the more first pass you encounter), good for large doses, duration controlled by base, anyone can do it/good for people who can’t give oral ;) ○ Disadvantages → poor compliance (i wonder why), leaking/insertion issues, slow absorption compared to oral or IV Mechanism of absorption of a drug from the rectal and vaginal route ○ Rectal The suppository is inserted and dissolved in the rectum where it melts and then penetrates through the lipid membrane and is able to enter the blood. It is taken up by the middle and inferior hemorrhoidal veins which drain into the common iliac vein which drains into the IVC Hemorrhoids caused by dilation of these veins Rectum has no buffers and neutral pH ○ Vaginal The cells of the vagina are covered by a thick mucous When a suppository is inserted, it dissolves and is able to travel through the mucous and into the bloodstream It is absorbed into the vaginal plexus which drains into the internal iliac veins, into the common iliac, and into the IVC Rectal, urethral, and vaginal product characteristics and types of bases ○ Vaginal → PEG/gelatin bases, round/ovular shaped, 3-5 g, can also be compressed tablets  ○ Rectal → can be fatty or water soluble bases, cylindrical w tapered end, 1-1.5 in, ~2g ○ Urethra Female → 5 mm diameter, 50 mm length, 2 g Male → 5 mm diameter, 125 mm length, 4 g Preparation storage and handling of suppositories ○ Preparation Double cast method → mix all drugs with a portion of base and pour evening about halfway up all molds then top off with more base. Let cool, then remelt and repour to evenly distribute all drug ○ Storage Glycero-gelatin suppositories usually packaged in well-closed screw-capped containers and stored < 35 degrees C Cocoa butter suppositories are usually packaged by wrapping individually into partitioned boxes to prevent contact and adhesion Light sensitive – wrapped in opaque material e.g. metallic foil Commercially individually wrapped in strip-packaging in foil or PVC-polyethylene Patient counseling on suppositories ○ Explain how to insert (after insertion lie still for 15 minutes and do not use the bathroom for at least an hour), signs of instability, how to store Describe the anatomy of human skin and how it influences transdermal drug absorption ○ Skin is made of three layers: corneum, dermis, subcutaneous ○ Bc the skin is thick, drug movement is slow ○ Drugs cross skin barrier via three methods Intercellular → moves between the cells Transcellular → moves through the cells Transappendigeal → moves via follicles or glands Describe the advantages and disadvantages of the transdermal drug delivery systems ○ Advantages → avoids first pass, avoids pain from injections, sustained release with infrequent dosing/potential for 0 order, easy termination, better compliance  ○ Disadvantages → adhesive issues/irritations, not good for drugs that need high blood volume, uncomfortable, cannot be cut, and has OD potential (drugs that “burst” and hairless skin = more permeable), drugs must be free and soluble in the system and solution, no more than 50 mg/day Explain what the differences are between the different transdermal systems ○ Ointments → occlusive and hydrating, made of oily base, and resists evaporation ○ Microneedles → 300-600 um in length to puncture corneum, and is usually painless and used for vaccines and macromolecules ○ Iontophoresis → positive current from device drives the positively charged drug out of device and into the body and vice versa, skin has isoelectric current at 4 so the SC must be negative ○ Phonophoresis → ultrasounds make small holes for fluids to flow through, sounds at a certain wavelength disrupt the lipid bilayer of corneum and create larger holes for molecules to go through List the reasons for the targeted delivery of drugs. ○ Used to optimize and widen therapeutic index by localizing its effect to target area Target towards organs, systems, cells, organelles, or molecules ○ Used for drugs with low solubility/absorption, short half life, low therapeutic index, to overcome barriers, drug is unstable via other methods, Compare and contrast active targeting to passive targeting with examples. ○ Active → drug is conjugated to carrier like monoclonal antibody to target antibodies at specific site or natural drug deposition of a carrier drug is altered, increase cellular uptake, fewer side effects and broader use, more challenging to synthesize Monoclonal antibody (no other example is given so this?) ○ Passive → physicochemical properties are altered and exploited, alters drug distribution in body, restricted in use, more side effects, and less effort to make Nanoparticle (no other example given so this?) Explain the general considerations in targeted drug delivery. ○ Target site → each tissue has things that can make delivery harder/easier - eye is easy bc you can inject into it whereas the brain is very impermeable  ○ Site specific properties → make sure you identify the properties different than other organs - ID proteins/antigens at that place only ○ Drug characteristics → keep in mind the physicochemical properties to determine method of delivery and final formulation ○ Target modality → must apply mathematical, scientific, and protein engineering to make final formulation - classify into particulate or conjugated drug systems Discuss the classification of targeted drug delivery systems (TDDS) with examples. ○ Generations First generation → normal pill things Second generation → enteric Third generation → swelling/diffusion controlled Fourth generation → target/self regulated delivery systems Fifth generation → gene therapy/biologics ○ Size Soluble macromolecules → prodrugs, mAb, immunotoxins Particulate carrier systems → liposomes, microparticles ○ These allow for the drug to reach their desired target and remain there for the required amount of time Identify the components of TDDS and their purpose. ○ Active drug ○ Carrier system for protection from metabolism and early clearance, and enhance favorable distribution ○ Homing system for specific targeting and not required for passive Describe the delivery challenges of various therapeutics and targeted delivery strategies. ○ Small molecules → BS Class 4, reduce off target toxicity ○ Proteins and peptides → improve stability and targeting, need non-invasive administration ○ Antibodies → achieving high doses, need non-invasive, bypass biological barriers ○ Nucleic acids → prevent off targeting gene alterations, accessing cytosol/nucleus ○ Live cells → control unpredictable PKs, manufacturing and scaling up, targeting disease location  Compare and contrast Microparticles and Nanoparticles. ○ Microparticles are 1-1000 um → red blood cells, polymeric microcapsules and particles ○ Nanoparticles are 1-1000 nm → DNA, virus, micelles Advantages → enhances solubility and therapeutic response, rapid drug degradation and bioavailability, lower side effects Describe different classes of nanoparticles and their advantages. ○ Polymeric → dendrimer, polymersome, nanosphere Easy modification and control of characteristics, has potential for aggregation, ○ Inorganic → silica and gold NP, quantum dot Unique and variable size and properties, good for theranostic applications, can be toxic and have solubility issues ○ Lipid based → liposome, emulsions Simple formulation with a range of properties, high bioavailability and flexibility, but low encapsulation efficiency Define liposomes and list their advantages and disadvantages. ○ Liposomes are sphere shaped vesicles with a lipid bilayer and can trap drugs in either layer for transport Comes in different types like SUV, LUV, GUV, multilamellar and multivesicular Explain the composition of liposomes and the function of each component. ○ Phospholipids → main component with hydrophilic heads and hydrophobic tails made of glycerophospholipids or sphingomyelins ○ Sterol → gives lipids fluidity, stability, and permeability made of cholesterol ○ Polyethylene glycol → protect metabolic inactivation and drug degradation to increase circulation time in body and enhance intracellular uptake made of DSPE-PEG ○ Charged lipids → allow for encapsulation of anionic drugs and better intracellular delivery made of DOTMA and stearyl amine Classify liposomes based on size, lamellarity, and composition.  ○ Conventional → basic and basis for all other liposomes, made of natural lipids or phospholipids with cholesterol used in passive targeting and unilamellar DaunoXome and AmBisome ○ Stealth aka long circulating → surface modification with PEGylation, avoid phagocytosis, the PEG binds to phospholipids in the body to allow them to circulate longer and have a more localized effect used in passive targeting Doxil and Onivyde ○ Immunoliposomes → designed for active targeting with specific antibodies on surface to improve targeting and appears unilamellar Compare and contrast Liposomes to Lipid nanoparticles (LNPs). ○ LNP → encapsulate R/DNA and are most clinically advanced non-viral DS, not all have a continuous bilayer as they have a monolayer with multilayer cores to help hold the drugs, they also do not have an internal aq pocket Advantages → optimized delivery, low immunogenicity, good targeting and encapsulation *** generally more stable after being lyophilized bc of long term storage Disadvantages → expensive and specific storage/formulation ○ Look at liposome lecture bc no new material was stated Describe nucleic acid therapeutics and the challenges associated with them. ○ 2 types of therapeutics mRNA based and synthetic oligonucleotide → have long lasting effects and can be used in replacement, editing, or inhibition, ASOs, siRNA → effective by disrupting transcription of DNA and translation of mRNA to proteins ○ Challenges Enzymes → attack added DNA and break it down Delivery → RNA systems are large, hydrophilic and anionic making it hard to get them in cell Off target → need specific targeting system to allow drug to reach target to decrease toxicity and increase efficiency  Getting to nucleus/cytoplasm → need to be able to be endocytosed so it can travel to target area Pharmacokinetics → naked nucleic acids break down easy and the body is too good at identifying and killing foreign DNA Physical instabilities → exposures at different temps, pH, transition metals, oxygen Chemical instabilities → hydrolysis, oxidation, deamination Discuss the typical composition of LNPs and the use of each ingredient. ○ Nucleic acid/drug ○ Cholesterol ○ PEG lipid → stealth/increase circulation (DMG-PEG) ○ Complexing lipid → to fuse with nucleic acid, membrane, endocyte (catatonic lipid) ○ Helper lipoid → stealth and structure (DSPA or neutral phospholipids) Explain the importance of cationic and ionizable cationic lipids in LNPs. ○ Catatonic → permanent + charge, commonly used are DOTMA and DOTAP, can rxn with nonspecific - charges which can lead to decreased hemolysis, cell adhesion, and transduction efficiency ○ Ionizable → + at low pH and neutral at physiological pH (neutral), this is good for in vivo bc neutral lipid have less interactions so they are more bioavailable, they have a pKa of 6-7 and are less toxic than fixed lipids Become + charged in endosome which can enhance release Ionizable LNPs are manufactured in acidic or low pH buffers (4-4.5) to allow the complexation of nucleic acids ONPATRO - IV (LNP siRNA), Spikevax and Comirnaty/Pfizer - IM (LNP mRNA) Identify different classes of polymeric nanoparticles. ○ Polymeric particles are colloidal with a size of 1-1000 um and are either made of natural or synthetic polymers → natural not approved by FDA due to batch to batch variations EXCEPT for albumin, all on market are synthetic ○ Types  Nanospheres → solid matrix, continuous polymer matrix with drug distributed within the matrix via encapsulation/covalent bonding or adsorption to nanosphere Nanocapsule → cavity surrounded by polymeric shell, used to encapsulate drugs in the liquid inner core Polymersomes → artificial vesicles, comparable to liposomes in shape and have good drug retention/stability, made of amphiphilic (self assembly) block copolymers, and biomimetic analogs (“copy”) of natural phospholipids, size 50 nm to 5 um, release drugs based on passive diffusion and stimulus reacted Micelles → made of the same copolymer and self assemble to have a hydrophilic shell w hydrophobic core and the drug goes into whatever layer its soluble in → this allows it to be used for IV, size of 10-200 nm, critical [micelle] which is the [] to keep the micelles assembled → fall below micelle fall apart Dendrimers → hyper branched polymers w a core and branches holding it out, easily control the mass, size, shape, chemistry, functional groups on outside allow conjugation in order to transport drugs (typically nucleic acids and small molecules) from blob to blob, typically for these we use charged particles like PAMAM and PEI ○ Advantages → more stable than lipids, biodegradable, easily manipulated properties and drug release, act as drug solubility enhancers ○ Disadvantages → risk for aggregation/toxicity, expensive, hard to scale up Discuss the mechanism of drug release from microparticles. ○ Creating a depot at injection site ○ After diffusion, drug will release slowly, the polymer will degrade overtime (after duration of use has passed) and then the whole thing will hydrolyze to be degraded and eventually eliminated Describe the importance of PLGA and PLA polymers in microparticles and nanoparticles formation.  ○ Their biodegradable and mechanical properties as well as MWS, and lactide:glycolide ratio (PLGA) are easily manipulated ○ Both are easily broken down by the body via Krebbs and have minimal toxicity associated with them ○ These are the only two polymers approved in the market PLGA is most successfully used bc it hydrolyzes into lactic and glycolic acid The more G you have the faster it degrades → need it to last longer? Add more L List the clinically relevant microparticle formulations and their composition and route of administration. ○ Lupron Depot → IM injection for prostate cancer formulated with PLA and PLGA for different durations of action ○ Bydureon → GLP-1 with PLGA as a subQ injection for diabetes ○ Risperdal Consta → PLGA IM injection every 2 weeks for schizophrenia Explain the methods of gene therapy. ○ In vivo → gene transferred into patient via vector, ocular therapy for congenital blindness ○ Ex vivo → gene put into isolated cells and then injected back into patient, modify the genome in vitro and insert back, focused on T or NK cells Like in vitro List the various gene therapy delivery systems. ○ Non viral → physical → gene gun, electroporation, micro injections AND chemical → polymers, liposomes Transfection technologies → transfer genes into cell using non-viral methods ○ Viral → biological → retro, adeno, and associated adenoviruses, do not cause disease bc their replication factors are removed and replaced with the genes we want to insert Transduction technologies → transfer genes into cells using viral vector  Integrating viral vectors → retro and lentivirus add genes to hosts DNA and is then replicated with the host Lentivirus and gamma retroviruses fuse fragment of their genetic material to host DNA, and are very stable and long lasting, both have moderate transduction efficiency, stable expression, and moderate/high immunogenicity ○ Kymirah and Yescarta are CAR-T therapies for blood cancer and are made by ex vivo integration of vectors Non integrating → adenoviruses add genes to host cell nuclei but do not integrate with DNA and have transient effect (short time) Adenoviruses have E1 region removed so it can’t replicate, is highly immunogenic and gene carrying abilities, high transduction efficiency and immunogenicity and transient expression ○ Janssen COVID-19 vaccine Adeno associated viruses → commonly used bc of their low toxicity and wide usage, typically does not integrate into genome, can only insert a limited amount of genes, moderate transduction efficiency, low immunogenicity, and stable/transient expression Luxturna and Zolgensma Discuss the physical methods employed in gene delivery. ○ Electroporation → use of physical force to insert drugs/macromolecules into extracellular compartments, transiently increases membrane permeability by disrupting it, simple, low cost, used in both in and ex vivo, but has short term pain, erythema (allergic rxn to meds), and tissue damage ○ Gene gun → particle bombardment → particles with genetic information are shot at a high velocity to impact and penetrate the membrane and transfect cells, requires specific particles and can cause cell damage ○ Sonoporation → transient pereation of membrane to add DNA to cell via ultrasound energy, noninvasive, combined with bubbles, but can cause cell and tissue damage and is not precise and hard to replicate  ○ Jet injection → another form of particle bombardment, minimal immune response, required training and is dedicated/time consuming Identify the chemical methods of gene delivery. ○ Liposomes/catatonic lipids → complex w nucleotides of interest and pass through cell membrane to release in cytoplasm ○ Cationic polymers → water soluble and form complexes with nucleic acids/polymers to adhered to cell membrane and endocytose ○ CaP precipitation → CaP binds to DNA on cell surface for endocytose ​Describe different classes of protein-based therapeutics with examples. ○ Antibody → produced by B lymphocytes to immune response and bind to antigens to help target certain cells Gazyva → mAb ○ Enzymes → catalyze rxns and replace a deficient/missing enzyme PEGaspargase → PEGylated L-asparaginase ○ Coagulation factors → naturally occurring proteins that help in blood clotting and can be admin to people w/o them SevenFact ○ Protein hormones → secreted by endocrine glands and act as messengers to stimulate a physiological cascade of action Insulin ○ Cytokines → broad group mediating cell - cell communication Interferon alfa b2 Identify the desirable characteristics of protein-based therapeutics. ○ Stability → proteins naturally break down easy so should not easily degrade, denature, aggregate ○ Targetability → want it in the body for longer time ○ Intracellular delivery → they also naturally have many targets in a cell so we have to tailor it ○ Pharmacodynamics → we want to increase/decrease kidney clearance through body w/o any extra side effects or immune responses Explain the role of PEGylation and Fc fusion in protein-based therapeutics.  ○ PEGylation extends half life of protein and decreases clearance ○ Fc fusion the Fc region of an antibody is fused to a biologically active protein which enhances circulation half life by decreasing the hydrodynamic diameter and slowing it down in the kidney (enhance circulation time by 1.4-1.8x) Discuss protein nanoparticles and their role in drug delivery with examples. ○ Advantages → increased stability and activity due to increased protection (so these next things decrease) from enzymatic degradation, immunogenicity, phagocytosis, and renal clearance, thereby leading to an increase in the half-life of the drug. ○ Ontak → targeted protein nanoparticle made of IL-2 and diphtheria, used for leukemia and lymphoma due to its IL-2 targeting, cytokines ○ Virus like particles → self assembled viral coat proteins, structure similar to viruses, but because of lacking nucleic acids they cant replicate, they are good carriers for therapeutic nucleic acids and other drugs Gardasil-9 ○ Human Serum Albumin → makes up > 60% of the blood plasma, globular protein, acts as drug carrier and enhances hydrophobic solubility in the body Paclitaxel-nab → contained in nanoparticles, binding this drug to albumin eliminates the need for solvents but can cause rxns, Abraxene used for breast, non small cell lung and pancreatic cancer List the components of antibody-drug conjugates and their function. ○ mAb → responsible to the selective targeting and binding ○ Cytotoxic or payload → must be high cytotoxic, stable, and have high potency after cleavage in target cell ○ Linker → connects antibody and payload and are typically microtubule disruptors like aurstatins or DNA damaging agents like doxorubicin Explain the mechanism of action of antibody-drug conjugates (ADC). ○ Bind ADC to specific antigen and the cell will internalize it ○ Degrading the ADC via lysosomes will release the payload ○ Cytotoxic effect occurs which is usually damaging DNA or disrupting microtubules  ○ The cell goes through apoptosis and dies List the physicochemical and biological attributes of nanomedicine products. ○ Formulation → how to the physicochemical properties affect the drug and release ○ Biological → how the drug interacts with biological systems and uptake (anaphylaxis) ○ Clinical → taking the medicine from the lab to the real world and focus on scalability, safety, and efficacy Discuss the role of particle size, surface properties, drug loading, and drug release in establishing the quality of nanomedicine products. ○ Particle size → key attribute as it affects many other things, first way to determine size is by dynamic light scattering, look at morphology via microscopy (TEM/SEM) ○ Surface properties → determines nature and interaction with external environment like proteins, biomolecules, and immune cells, things such as SA, ligands and hydrophobicity of particle surface affect this ○ Drug loading and release → encapsulation helps to ensure desired therapeutic effect and drug safety, measurement of total drug loaded is achievable by dissolution of vehicle and extraction/quantification of drug released, if the nanoparticle is unstable/porous it can all diffuse out into the blood and cause adverse effects Explain the kinetic properties of nanomedicine products in biological media. ○ Must be biologically stable under three conditions at different temps and pHs to see how they react in different body conditions such as fever/inflammation Release kinetics in presence of plasma proteins - shows impact on safety/efficacy and measures drug release Physical stability of nanocarrier regarding change in size and polydispersity of particles when contacting plasma proteins - measures stability in the body and doesn't work on small “soft” molecules Adsorption of plasma proteins on the drug surface leading to protein corona and alters surface properties - measures interaction with immune system as well as safety/efficacy and doesn't work on small molecules -  nanoparticle “grows” in body bc the proteins adsorb to it forming the corona Identify the cellular responses and adverse effects associated with nanomedicine products. ○ Systemic administration → RBC lysis = haemolysis ○ NP surface opsonization/protein adsorption → platelet aggregation =thrombogenesis, complement proteins = CARPA, proinflammatory cytokines and ROS production = inflammation, and antiinflammatory cytokines = immune response Describe the importance of endotoxin contamination and hemocompatibility in evaluating nanomedicine products. ○ Endotoxin → LPS that are parts of a gram - cell wall, contamination can occur in manufacturing/handling, in vivo testing with LAL assay (gel clot, turbidimetric, and chromogenic) ○ Hemocompatibility → assessment includes studying hemolysis and thrombogenesis testing, in vivo hemolysis assay is a good indicator on whether your particle can be used for this purpose Explain the concept of complement activation-related pseudoallergy (CARPA) in the context of nanotechnology-based health products. ○ Hypersensitivity rxn to show mild to severe cardiopulmonary issues and is required for nano medications ○ Anaphylaxis is not IgE mediated bc it is triggered by complement activation → activating this system in vivo to evaluate CARPA risk for people Explain the differences between machine learning and artificial intelligence ○ ML → branch of AI in computer science - data sets/stats/boring shit (like evan) In real world we have tablets with info on patients and its connected to data and computations done in a database and the “are u human” ○ AI → field of study to mimic how humans learn to predict - mimic how we think and learn Discuss data curation ○ Organizing and managing data sets to find and meet needs of a certain group of  people ○ Steps → obtain data sets, clean and validate, experiment to develop new models, test models on new data sets, apply to real world Explain the differences between training, test and validation sets, data find and data mining\ ○ Training set is used to train the program on how to sort and give the desired answers ○ Test sets test the program on how well it runs on an entirely new data set and validation sets validate the final models for ○ Data find is …… idk command F???? ○ Data mining is math based and is able to reveal patterns or trends in data Explain what is meant with “black box” ○ A proprietary system with an algorithm to sort data to come up with relevant information, when picking a program vendor to use make a test set of data and run it to see which operating system works best as we do not know how the black box sorting works Identify websites, programs which can be used for ML/AI ○ Programs → Python, MATLAB, Amazon Web Service, orange ○ Operating systems → software that manages the hardware while providing an environment for the systems/programs to run macOS, Windows, Linux Explain role of wearable/phone devices on medical health and information, in context of patient education ○ They allow patients to have easy access to basic information about their current health ie. HR, BP, daily movement, glucose monitors (FreeStyle Libre), etc ○ They are able to raise concerns about their health and communicate this with their providers who can validate their information and give them the help they may need Study Guides What is a biosimilar?  ○ A biological medication that is similar to an already FDA approved biological product (the OG is the innovator biologic) What different types of biopharmaceuticals are identified? ○ Types of biopharmaceuticals Monoclonal antibodies Enzymes Immunomodulators → affects immune response Cytokines → impacts cell to cell interaction/communication What is freeze drying of proteins? ○ AKA lyophilization ○ Remove water from a protein sample in order to turn it into a powder to obtain a longer shelf life What types of instability can occur with proteins? ○ Chemical and physical → read above for more information What is surface adsorption? ○ When the drug/peptide sticks to the wall of the container it is in. It is seen as a sign of instability as it can lead to protein degradation. What protein used in type I diabetes tends to adsorption to plastic for IV tubing? ○ Insulin What temperature is a freezer and refrigerator? ○ Freezer = -20 degrees, fridge = 2-8 degrees What peptide of drug is orally available? ○ Cyclosporine → available in a solution and soft gel What nasal spray peptide can be used for osteoporosis? ○ Miacalcin Afrezza → insulin Desmopressin → diabetes Nafarelin → endometriosis What peptide drug is used in induced labor after pregnancy? ○ Pitocin → it’s an artificial form of oxytocin to induce uterine contractions What is an antibody drug conjugate?  ○ A monoclonal antibody combined with a cancer drug used to target certain cells ○ The antibody binds to the cancer cell and the medication is able to then attack the cell ○ A monoclonal antibody is a clone of a WBC to function in immune responses Where are the main delivery sites of suppositories? ○ Vagina, rectum, urethra What are vaginal suppositories mainly compounded as base? ○ Made of water soluble bases like PEG and gelatin to minimize leaking ○ Be careful with Na barbital, salicylic acid and camphor will crystalize out of PEG Which suppositories are bougie and pessaries? ○ Bougie is the urethral suppositories and the pessaries are the vaginal What are the main uses of suppositories, ie. Local and systemic? ○ Local → hemorrhoids, infection ○ Systemic → hormones, anti-nausea, analgesic What effect does suppositories have on first pass metabolism? ○ The lower part of the rectum, ya know where evan gets pegged, avoids first pass and the things absorbed here enter systemic circulation as the middle and inferior rectal veins drain straight into the IVC. The further up the rectum you go the things absorbed here are transported to the liver via the portal system which would then go through first pass. What is the influence of suppository based on drug release? ○ Water soluble bases will dissolve easier in the body and therefore be absorbed faster due to them being miscible with body fluids. FA bases will still melt quickly but they will take a longer time to make their way into the blood due to their nature. Which suppository base has problems with polymorphism? ○ Cocoa butter → if it's not heated/cooled correctly you could get the incorrect polymorph of the butter What effect does glycerol suppositories have as a possible side effect or therapeutic effect? ○ The major side effect to be aware of is that glycerol can have a laxative action  ○ However, this means it can used to treat you when ur full of shit What excipients can be used in suppositories? ○ Antioxidants → BTH, BHA, the “gallates” ○ Preservatives → benzoic acid, cetrimide → most bases will required one ○ Emulsifiers → wool fat/alcohol, polysorbates → used to facilitate incorporation of an aq liquid ○ Hardening Agents → gelatin Where should cocoa butter and fatty base suppositories be stored compared to PEG ones? ○ FA ones are individually wrapped and packed in a box to store in a fridge or at least below 30 degrees, but NOT IN FREEZER ○ PEG are put in a tight screw cap container and stored below 35 degrees What are instabilities of suppositories? ○ Brittleness → caused by lack of humidity ○ Sponginess → caused by high humidity ○ Excessive softening → most important instability sign What are the layers of the skin that prevent drug delivery? ○ The corneum prevents delivery as it is mostly dead keratin cells which do not support facilitation and absorption of drugs as there’s no blood flow What would happen to drug absorption if the A or h is increased/decreased with Fick’s law? ○ A is a surface area of the membrane, and by increasing surface area we will increase drug absorption. The letter h represents membrane thickness, so by increasing it we will decrease drug absorption. What is a PLO gel? ○ A lecithin based organogel used as a vehicle in compounding to help improve permeability of many drugs. What allergy might occur with PLO gels? ○ It contains egg and soy What types of patches can be made, e.g reservoir etc? ○ Adhesive device → a backing (aluminum/polyester film) and a drug containing adhesive  ○ Monolithic device → a backing, adhesive (acrylic polymer) , and rate controlling drug containing membrane ○ Reservoir device → a backing, adhesive, rate controlling membrane (porous polypropylene), and drug containing reservoir (mineral oil/polyisobutylene) They all have a peel off layer to adhesive made of siliconized polyester What would the following properties have on drug candidates for TTDS: logP and MW 1 and < 4 for optimal passive drug delivery What factors would enhance drug absorption via transdermal systems? ○ We would want things such as permeation enhancers in order to better penetrate the corneum layer of skin ○ Works by altering hydration, intercellular lipids, reducing corneum resistance ○ DMSO, propylene glycol, ethanol, surfactants What would DMSO or surfactants do for drug penetration through skin? ○ DMSO helps with disrupting the skin lipid bilayer and solubilizing the drugs in order to help them better absorb. ○ Surfactants help to emulsify the drugs in the delivery system and breach the gap between possible hydrophilic/hydrophobic properties of drugs/body/systems. What would happen if a patent cuts a patch? ○ By cutting a patch you risk affecting the sustained release properties and the drug may release all at once causing potential OD Other important notes Do not use microparticles for IV → will cause embolism → if you see the word “depot” it will never be IV ○ Micro are good for long acting injectables due to their sustained release capability GLP-1 injections should be given once every seven days for diabetes PGLA lasts 1 month, whereas PLA last 3, 4, or 6 months Transdermal drug delivery  ○ Liberation on skin surface, penetration through corneum, permeation into dermis, and finally absorption/diffusion into blood vessels Drugs available for TDDS ○ Good – Contraceptives, fentanyl, nicotine, testosterone ○ Bad - Valacyclovir is a prog drug Factors affecting transdermal absorption ○ [drug] → more drug, more absorbed ○ LogP → high = less aq soluble to affect bioavailability ○ Skin hydration → the more hydrated the better absorbed ○ Thickness or corneum → inversely proportional ○ Surface area applied to → proportional ○ Duration of contact w skin → proportional Classifications of drug delivery ○ First order → delivers drugs to capillary bed of target organ ○ Second order → deliver drugs to certain cell types sparing healthy cells ○ Third order → deliver drugs to intracellular locations Ideal characteristics of a targeted drug delivery system (TDDS) ○ Specifically target the drug to target cells or target tissue ○ Ensure minimal drug leakage during transit to target ○ Protect the associated drug from metabolism ○ Retain the drug at the target site for the desired time ○ Facilitate the transport of the drug into the cell ○ Deliver the drug to the appropriate intracellular target site ○ Be biocompatible, biodegradable, and non-antigenic Advantages and disadvantages of liposomes ○ Can deliver hydrophilic and phobic drugs, nontoxic and readily bioavailable, help avoid contact of hazardous drugs with sensitive tissues, and increased drug stability due to encapsulation ○ They are expensive, have a short half life, low solubility in water, and can leak/fuse leading to rxns  Protein therapeutics are good because they are versatile, acting as catalysts, signaling molecules, molecular and ion transporters, scaffolds for maintaining cellular and tissue integrity, receptors, and drug carriers. ○ They are less likely to cause side effects by interfering with normal biological processes because they have evolved to play highly specific roles. ○ Typically, protein therapeutics show high potency and can also execute more complex functions owing to their intricate three-dimensional structures. Select a viral vector → type of target cell, packaging/stability, and desired duration of gene expression ○ Viral is good for ex vivo and transplantable stem cells ○ AAV is good for in vivo vehicles The ionizable lipid nitrogen: oligonucleotide phosphate (N/P) molar ratio represents the charge balance between the cationic tertiary amine of the ionizable cationic lipid and the anionic phosphate group in the oligonucleotide backbone. This property is the basis for the complexation of ionizable cationic lipids with oligonucleotides. LNPs commonly have an N/P ratio of around six (6). To understand the absorption, distribution, metabolism and excretion (ADME) profile of product under investigation it is important to use a range of model systems from in vitro cell-based models to in vivo animal models. ○ All assays require the detection and quantitation of nanomaterials in biological matrices, such as tissues, cells and subcellular structures