Flashcards in Late Stage Drug Development Deck (53):
1
what do you use to test for intestinal absorption>
a Caco-2 cell monolayer
2
if a drug has stability against CYP450, less than ____ metabolism to the drug will occur after 1 hour
50%
3
for in vivo metabolic stability studies, greater than _______ percent of the drug should remain after 1 hour in rodents
20
4
general cytotoxicity studies include
ATP measurement, MTS, enzyme release
5
to check for mitochondrial dysfunction:
look at oxygen consumption
6
hERG potassium channel inhibition can cause
life-threatening arrhythmias
7
drug-drug interactions are usually _________ related
metabolism
8
paracellular and transcellular are forms of ____ uptake
passive
9
carrier mediated uptake is a form of ____ uptakes
active
10
Efflux is mediated by __________
p-glycoprotein
11
Caco-2 cells exhibit __________ mechanisms
all forms of uptake and efflux
12
the small intestine side of a membrane is called the _______ side
apical
13
the blood side of a membrane is called the __ side
basolateral
14
how many layer of cells are used in Caco-2 testing
one, a monolayer
15
hepatocytes are isolated from ____ and used as _______ cells. They contain the entire range of metabolic enzymes. A downside is that they are ____
animal/ intact/ harder to grow and use
16
microsomes:
subcellular fraction of digested liver
17
microsomes consist of ________
endoplasmic reticulum
18
microsomes contain ____ enzymes only
phase I
19
____ and ______ are phase I enzymes
CYP450/ FMO
20
microsomes are _____ to use than hepatocytes
easier
21
metabolism stability: half life should definitely be over _______, and _____ would probably be good to proceed to the next step
10 minutes/ 20 minutes
22
To do stability tests: start with a 96 well plate. Add ________. Then add ________. Now _________. Then ________ and ______ --> quantification of parent compound disappearance
chemicals/ hepatocytes or microsomes/ incubate/ extract/ analyze
23
to see if a drug inhibits CYP450, add ___, then ____ and incubate. Then add ____, incubate, and check for ______.
chemicals/ hepatocytes/ P450 substrate/ metabolites
24
toxicity can be measured as hepatocyte viability in the presence of the drug using _____
MTS/ MTT colorimetric assay
25
name three cytoplasmic enzymes that can be tested for to analyze the amount of cell death
lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase
26
_____ percent of the cell's energy is produced in the mitochondria
90
27
where is the programmed cell death control center located?
in the mitochondria
28
what are three main screens for mitochondrial toxicity?
disrupted energy metabolism, increased free radical generation, altered apoptosis
29
name four screening assays to check for energy metabolism disruption
ATP production (reduction = toxicity), membrane potential, oxygen utilization, mt biogenesis
30
name a screening assay that checks for free radical production
ROS production
31
name a screening assay that checks apoptosis induction
PARP activation
32
name three mechanistic assays that check energy metabolism disruption
OXPHOS Enzyme activity, phospo + acetyl, metabolic protein expression panel
33
name three mechanistic assays used to check for free radical production
protein nitration panel, protein oxidation panel, antioxidant protein expression panel
34
name two mechanistic assays use to check for apoptosis induction
cell fractionation and protein translocation panel, apoptosis protein expression panel
35
normal heart rhythms are governed by
electrical pulses (based on ion transport)
36
disruption of the electrical pulses that regulate heart rhythms results in
prolonged QT interval (prolonged heart beat) eventually arrythmias and possibly death
37
the hERG potassium channel is the major protein responsible for conducting K+ ions ____ the heart muscle cells
out of
38
name two drugs that are hERG inhibitors
fluroquinolones, terfenadine
39
what test can you use to check for genotoxicity?
Ames test
40
You need ___________, _____________, ________ and ____ studies to open an IND
proof of principle in most appropriate animal model, acute and chronic toxicity studies, safety pharmacology (respiratory, CVS, CNS), ADME panels
41
acute toxicity studies:
one large dose or several large does given over less than two weeks
42
chronic toxicity studies:
should be for the length of the clinical trial
43
therapeutic index:
LD50/ED50
44
phase I trials provide ___________ and determine _______
initial pharmacologic and pharmacokinetic data/ maximum tolerable dosage (MTD)
45
initial dose is usually _____ for phase I trials
1/10 of MELD10
46
______ ______ volunteers are used in a phase I trial
20-100/ healthy
47
________ interactions studies are performed in phase I trials
drug-food
48
how long are drug patents for?
20 years
49
four main criteria for patentability:
novelty, inventive step, enablement, support (justification for claims of efficacy)
50
patent claims are written to cover ____
a wide range of compounds in a given class (scaffold)
51
terfenadine is a prodrug _____ converted to ______-- by ________. It is a _______.
antihistamine/ fexofenadine (Allegra)/ CYP 450 3A4/ hERG channel inhibitor
52
name four hERG channel inhibitors
HIV protease inhibitors, Gleevec (cancer drug), ketoconazonle, grapefruit juice
53