Problem Discussion

1. a. Feridex® and Ferumoxytol are two different iron oxide-based MRI contrast agents. The DLS size of Feridex® is
120-180 nm and that of Ferumoxytol is 30 nm. Between these two formulations which one will you prefer for – i)
liver imaging; ii) metastatic lymph node imaging; iii) cellular imaging and iv) cerebral blood volume MRI?

Ans. Feridex® - Liver imaging (HD size > 100 nm, so nanoparticles will be engulfed by liver macrophages.
Ferumoxytol – very small HD and coated with carboxymethyl dextran based coating. Very long intravascular half-life
(10-14 h). So this can be used for vessel imaging and neuroimaging applications including brain angiography.

b. Abdoscan® - oral route

c. i) T2 - weighted contrast agent because the post-contrast image is darker.
iii) Magnetic resonance imaging (MRI) differentiates between body tissues by measuring the time it takes for protons
to realign with a magnetic field, and the amount of energy released. The time it takes for protons to realign is called the
relaxation time, and there are two types: T1 and T2. Different tissues in our body have different relaxation times, which
allows physicians to identify them.
 •A and B represent the in vivo hippocampal R2* signal changes of a rat brain administered with two different variants of carboxymethyl dextran-
coated iron oxide nanoparticles: CION-A and CION-B. Both contrast agents were administered 15 min after the onset of the MRI scan and the
changes in contrast signal as a function of time are shown below. What would be your inference about the pharmacokinetic profile of CION-A and
CION-B? Based on their pharmacokinetic profile, which of these two contrast agents would you think is preferable for cerebral blood volume (CBV)-
weighted functional MRI (fMRI) and high-resolution MR angiography?

B 120

100

delta R2 * (s-1)
80

60

40

20

0
1 16 31 46 61 76 91 106 121 136 151 166 181 196 211 226 241
Time (min)

Very fast clearance from the blood Long intravascular half-life – can be
used for prolonged imaging applications
Magnetic resonance angiography (MRA) is a type of MRI that including MR-angiography
uses radio waves, magnetic fields, and a computer to examine
the body's blood vessels.
 Preclinical Applications of CION

The blood-oxygenation-level-dependent (BOLD) signal

Cornerstone of modern neuroimaging
Breath pure oxygen Breath normal air

• Deoxygenated hemoglobin is paramagnetic and thus creates negative contrast.

Ogawa et al., Magn Res Med, 1990 3

 Cerebral blood volume (CBV) Mapping

BOLD-fMRI has poor SNR, low sensitivity and spatial specificity

MR signal ↓
CBV ↓
Neuronal Firing
MR signal ↑

Blood Flow increases

CBV ↑
vessel MR signal ↓
Injection of iron oxide
nanoparticles Vessel dilates
MR signal intensity

Steady state CBV changes

Stimulation
Time BOLD signal
correlates with CBV
changes
CBV fMRI produces higher contrast-to-noise ratio compared to BOLD-fMRI
 MRI Contrast Agents
- Improve visibility of internal body structure in MRI

Non-contrast Contrast-enhanced Pre-contrast Post-contrast

MRI Contrast Agents

T1- weighted: T2- weighted:

Eg. Gadolinium- Eg. Iron oxide
3D MIP images obtained from based chelates nanoparticles Human brain vascularity
2D TOF MRV (A), and CE MRV at the level of the basal ganglia 2
1

• Positive contrast: • Short intravascular • Negative contrast: • Size and surface

Post-contrast image half life. Not ideal for Post-contrast chemistry can be
looks brighter blood pool imaging image looks darker tailored to render
long-circulating
• Alter T1 relaxivity of • Causes renal toxicity • Alter T2 relaxivity of properties
surrounding water surrounding H2O
molecules • Gd deposition in brain molecules • Low toxicity in vivo

5
http://www.ajnr.org/content/30/1/185 http://www.ohsu.edu/xd/research/centers-institutes/airc/research/index.cfm
 Basic Principles of MRI
When a person goes inside the MRI scanner, the
randomly oriented protons inside the water molecules
of their body align themselves with the direction of the
main static magnetic field, B0. Some of these protons
will align “up” or parallel whereas others will align
“down” or antiparallel, while still rotating around their
axis like a spinning top.
B0

All living bodies are made up of

water molecules, constituting
two hydrogen atoms and one
oxygen atom. Each hydrogen Step I: Step II: Step III: Step IV:
nucleus contains one positive All protons RF excitation Aligned protons RF pulse is turned off
charge i.e., a proton spinning become aligned pulse is applied are excited by and the net
around on its axis behaving like a with the net to the aligned RF pulse and magnetization begins
magnetization B0 protons flips into the to relax and return to
tiny magnet. transverse plane the longitudinal plane
MRI Relaxation
• T1 is a measure of the time taken by the excited
protons to return to equilibrium and realign with the
external magnetic field.

• T2 is a measure of the time taken for the spinning

protons to lose phase coherence among the nuclei
spinning perpendicular to the main field.

• Depending on their cellular and molecular

characteristics, different tissues have different T1
Longitudinal Relaxation or Transverse Relaxation or and T2 relaxation times, which forms the basis of
Spin-lattice Relaxation (T1) Spin-Spin Relaxation (T2) image contrast.
Which imaging modality do you think will be better for diagnosing Alzheimer’s disease: MRI or PET? Explain briefly.

Brain scans known as fluorodeoxyglucose PET, also called FDG PET, may be used to diagnose Alzheimer's disease. FDG
PET scans show areas of the brain where nutrients aren't properly used for energy, known as metabolism.

The major strength of MRI is its widespread availability to be used in AD research. MR images provide excellent
anatomical detail, and hippocampal atrophy measured on a high-resolution T1-weighted MRI serves as a critical
criterion for the clinical diagnosis of AD.

Hippocampal atrophy is a decrease in the size of the hippocampus, a part of the brain that plays a key role in
learning and memory. It can be a marker of Alzheimer's disease (AD) and other conditions, and it's associated
with cognitive decline
 Positron emission Tomography
PET/CT PET / MR

Positron emission tomography (PET) is a nuclear

medicine-based functional imaging technique that is
used to observe the metabolic process in the body.
Selection of modality depends on the question we want to address-

Modality Advantages Limitations

MRI, CT -Provides morphological information -Limited value for monitoring

such as size and localization of outcomes of therapy
tumors

- Delineates secondary phenomena

such as mass effect, edema,
hemorrhage, necrosis
PET - Characterize tumor on metabolic - Only functional information
and molecular level
- Not much structural overview
- More sensitive than MR and CT
(works in the picomolar range)
 Principles of PET imaging
Radioactive drug or tracer
18 F, 15 O,11C, 13N etc.

Wait time

1
Line of
response

Line of
response

511 keV

511 keV

1https://wiki.engr.illinois.edu/display/BIOE414/PET+Scanner+Overview
In PET, biochemically active molecules are labeled with short-lived positron-emitting radionuclides (radiotracers)
• Patients injected with radioactive drug or tracer that traces a specific pathway of interest
• Proton-rich isotopes may decay via positron emission, in which a proton in the nucleus decays to a neutron, a
positron and a neutrino. The daughter isotope has an atomic number one less than the parent.
• After traveling a short distance within the tissue positrons emitted from the nucleus of radioisotopes
annihilate on contact with electrons
• Each annihilation results in a pair of 511 keV photons traveling in opposite directions
• Emitted у rays are counted by detector units commonly arranged in a ring surrounding the
subject.
When 2 opposing detectors simultaneously register a pair of photons, the annihilation event is counted and
assigned to a line of response joining the 2 relevant
 Radiotracers in PET

F-18 labeled Fluorodeoxyglucose (FDG) is a glucose

analog that is taken up by high glucose-consuming cells

FDG-PET to map human brain’s response to different task

Visual cortex Auditory cortex Frontal cortex Hippocampus Motor cortex

PNAS USA 2000; 97(16): 9226-9233

 FDG vs FLT-PET
18F-FDG vs 18F-FLT-PET in Differentiating Radiation
Necrosis From Recurrent Glioma in same patient-

• FDG – Glucose metabolism. Normal gray matter

tissue has high glucose metabolism – False
positive interpretation

• FLT –(fluorothymidine) DNA synthesis/cellular

proliferation. Normal brain has low signal.
Clin Nucl Med 2012;37: 854-861
What does a low volume of distribution of a drug imply?

A drug with a low Vd has a propensity to remain in the plasma meaning a lower dose of a drug is required to
achieve a given plasma concentration. (Low Vd -> Less distribution to other tissue). A high VD value indicates
that the drug is distributed more in tissues than in plasma.

The average steady-state plasma concentration after administering an 800 mg sustained-release theophylline
product every 24 hours to a 78 kg male subject (5 feet 2 inches) is 15 ug/mL. The volume of distribution is
40 L. What is the drug’s approximate half-life?

Steady-state concentration
(Css) occurs when the amount
of a drug being absorbed is the
same amount that's being
cleared from the body when
the drug is given continuously
or repeatedly
A patient with an adverse reaction to Drug X requires that the plasma drug concentration not ever exceed 40
mg/L. If Drug X has a half-life of 8 hrs and clearance of 4 L/hr, what do you think is the best i.v. loading dose
that will give the maximum blood levels while avoiding the adverse reaction?

In the brain, endothelial cells and reinforcing astrocyte cells restrict the level of pinocytosis through the
formation of tight junctions between cells at the blood-brain interface. This results in the formation of a structural
and metabolic barrier, commonly referred to as the blood-brain barrier (BBB) – Can you suggest a strategy by
which a nano-drug delivery system can overcome the BBB?
“Lactoferrin is a multifunctional glycoprotein, which belongs to the transferrin family, and possesses
various biological functions such as antimicrobial, antibacterial, antiviral, antiparasitic, and
antitumor activities.22 Furthermore, low-density lipoprotein receptor-related protein (LRP), a
specific lactoferrin receptor with excellent selectivity, is highly expressed in glioma
cells.23,24 Lactoferrin is also able to cross the blood–brain barrier (BBB) through the transcytosis
processes within brain capillary endothelial cells.25 Because of their ability to target glioma cells and
cross the blood-brain barrier (BBB), lactoferrin-conjugated nanoparticles have great potential for
brain tumor targeting.25,26 Previous studies also have shown that lactoferrin-conjugated
nanoparticles have no effect on cell viability.23””

https://pmc.ncbi.nlm.nih.gov/articles/PMC4626448/
Following i.v. administration, NPs are cleared up within minutes from the bloodstream – What do you
think will be their final biodistribution?

Following i.v. administration, NPs are cleared up within minutes from the bloodstream and their typical
final bio-distribution is around 60–90% in the liver, 5–8% in the spleen, and 1–2% in the bone marrow.

Lisinopril is an orally active ACE inhibitor used to treat hypertension, heart failure, and heart attacks
Explain stepwise how Lisinopril will reduce blood pressure in hypertensive patients. Instead of Lisinopril, if
the doctor prescribes Nifedipine, will there be any difference in the mechanism of action? Explain.

Levocetirizine is an H1-receptor antagonist used to treat symptoms associated with chronic allergic rhinitis and
uncomplicated cases of chronic idiopathic urticaria.

The oral bioavailability of paclitaxel and docetaxel are 1% and <10%, respectively, whereas for doxorubicin
the value is <5% - Explain why most anticancer drugs have poor bioavailability. Suppose you are a part of the
formulations team in a leading pharmaceutical company that specializes in oncology medicines. Your team
leader has given you the responsibility to design a nanoformulation that can enhance the oral bioavailability
of a BCS Class IV anticancer drug. What type of nanocarriers would you propose to use?
Why do generic drugs often cost less than brand-name drugs?

A prescription drug that has the same API as a brand-name drug. Generic drugs usually cost less than
brand-name drugs. The Food and Drug Administration (FDA) rates these drugs to be as safe and effective
as brand-name drugs. Generic drugs tend to cost less than their brand-name counterparts
because generic drug applicants do not have to repeat animal and clinical (human) studies that were
required of the brand-name medicines to demonstrate safety and effectiveness. So, companies
developing generics don't have to invest major amounts of time and research because FDA testing and
approval of the brand drug's ingredients is already complete. That means generics can get to market
quicker and be sold for much cheaper than brand drugs

Name an FDA-approved orphan drug that was discovered and designed using Artificial Intelligence.

Codeine, a weak narcotic pain reliever and cough suppressant, is a prodrug that must be metabolized to
its active metabolites, morphine and codeine 6-glucuronide, to be effective. Cytochrome P450 2D6
(CYP2D6) is an enzyme that converts codeine into morphine to provide its analgesic effect. Based on
your understanding of ADME, explain whether the genetic variations in CYP2D6 will impact the
pharmacology of codeine across patients.
Surface plasmon resonance (SPR) is an optical technique that measures molecular interactions in real time. It
occurs when light hits a metal film at a specific angle, causing electrons in the metal to become excited and
travel parallel to the film. Surface plasmon resonance occurs when a metal is placed on the top of surface,
where light is undergoing total internal reflection. A group of electrons in the metal absorbs the energy from
the light undergoing total internal reflection. As a result of energy absorption, the group of electrons starts
oscillating. This group of electrons is known as plasmon, and the phenomenon is known as surface plasmon
resonance. Because of SPR, a dark zone appears in the refracted light.

SERS can be used to study SPR

SERS, or surface-enhanced Raman spectroscopy, is a technique that enhances the Raman scattering of
molecules that are near nanostructured metallic surfaces