Conditions: Alzheimer Disease
Keywords: Tau Amyloid MK-6240 Florbetaben Racial/Ethnic Minority Alzheimer's disease
Allocation: NA | Intervention model: SINGLE_GROUP
Enrollment: 145
Healthy volunteers: yes

Brief summary:
The study employs tau positron emission tomography (PET) imaging in a well-characterized multi-racial/ethnic cohort to examine the extent to which tau pathology is associated with cognition, differences in tau pathology across racial/ethnic groups, and the relationship between MRI markers of small-vessel cerebrovascular disease and tau pathology. The study also investigates amyloid-dependent tau spreading.

Detailed description:
Deposition of hyperphosphorylated tau protein is observed in several neurodegenerative diseases including Alzheimer's Disease (AD), progressive supranuclear palsy, corticobasal degeneration, chronic traumatic encephalopathy, and frontotemporal lobar degeneration. Tau is a microtubular protein and its native function is to provide structural support to neurons. Paired helical filaments composed of dysfunctional tau protein are found in several neurodegenerative diseases. In AD, the clinical progression of dementia has been shown to correlate with the amount and topographical spread of tau throughout the brain. Therefore, detecting and quantifying tau aggregate load in brain would have diagnostic and prognostic potential in clinical management of several neurological diseases. As disease modifying drugs that target tau are being developed, there is a critical need for a reliable method of detecting tau aggregates to confirm pathology in patients entering clinical trials.

Arms:
arm 1: Racially/ethnically diverse subjects with or without a positive family history of Alzheimer's disease (AD) will have one PET scan with 18F-MK-6240 over a 30 to 60-minute scanning period, and one PET scan with 18F-Florbetaben over a 20-minute scanning period.

Interventions: intervention 1: 18F-MK-6240 intervention 2: 18F-Florbetaben
Intervention descriptions: intervention 1: Administration of 5 mCi of 18F-MK-6240 for tau PET. intervention 2: Administration of 8.1 mCi as a slow single intravenous bolus (6 sec/mL) in a total volume of up to 10 mL of 18F-Florbetaben for Aβ PET imaging.

Protocol document:
Prot_SAP_000.pdf: 
ALZHEIMER’S PET IMAGING IN  
RACIALLY/ETHNICALLY DIVERSE ADULTS
Regulatory Sponsor: 
Adam M. Brickman, Ph.D. 
Taub Institute for Research on Alzheimer’s Disease & the 
Aging Brain 
Funding Sponsor: 
Study Product: 
NCT number: 
IND Number: 
National Institutes of Health/National Institutes on Aging 
9000 Rockville Pike 
Bethesda, Maryland 20892 
+1 (301) 496-4000
18F-MK-6240 
18F-Neuraceq, 18F-Florbetaben 
NCT03706261 
140399 
Protocol Date: July 29, 2022  
CONFIDENTIAL 
This document is confidential and the property of Columbia University Medical Center.  
No part of it may be transmitted, reproduced, published, or used by other persons without 
prior written authorization from the study sponsor. 


Page ii 
Table of Contents 
STUDY SUMMARY ................................................................................................................................................... 1 
INTRODUCTION ....................................................................................................................................................... 3 
1.1 
BACKGROUND .................................................................................................................................................. 3 
1.2 
INVESTIGATIONAL AGENT................................................................................................................................ 3 
1.3 
PRECLINICAL DATA ......................................................................................................................................... 3 
1.4 
CLINICAL DATA TO DATE ................................................................................................................................ 6 
1.5 
DOSE RATIONALE AND RISK/BENEFITS .......................................................................................................... 10 
2 
STUDY OBJECTIVES .................................................................................................................................... 13 
3 
STUDY DESIGN .............................................................................................................................................. 14 
3.1 
GENERAL DESIGN .......................................................................................................................................... 14 
3.2 
PRIMARY STUDY ENDPOINTS ......................................................................................................................... 15 
3.3 
PRIMARY SAFETY ENDPOINTS ........................................................................................................................ 15 
4 
SUBJECT SELECTION AND WITHDRAWAL.......................................................................................... 15 
4.1 
INCLUSION CRITERIA ..................................................................................................................................... 15 
4.2 
EXCLUSION CRITERIA .................................................................................................................................... 15 
4.3 
SUBJECT RECRUITMENT AND SCREENING ...................................................................................................... 16 
4.4 
EARLY WITHDRAWAL OF SUBJECTS ............................................................................................................... 17 
4.4.1 
When and How to Withdraw Subjects .................................................................................................. 17 
4.4.2 
Data Collection and Follow-up for Withdrawn Subjects ..................................................................... 17 
5 
STUDY DRUG ................................................................................................................................................. 17 
5.1 
DESCRIPTION ................................................................................................................................................. 17 
5.2 
PREPARATION AND ADMINISTRATION OF STUDY DRUG ................................................................................. 18 
5.3 
PACKAGING.................................................................................................................................................... 18 
5.4 
RECEIVING, STORAGE, DISPENSING AND RETURN .......................................................................................... 18 
5.4.1 
Receipt of Drug Supplies ..................................................................................................................... 18 
5.4.2 
Storage ................................................................................................................................................. 18 
5.4.3 
Dispensing of Study Drug .................................................................................................................... 19 
5.4.4 
Return or Destruction of Study Drug ................................................................................................... 19 
6 
STUDY PROCEDURES .................................................................................................................................. 19 
6.1 
TAU PET ........................................................................................................................................................ 19 
6.2 
AMYLOID PET ............................................................................................................................................... 19 
6.3 
QUESTIONNAIRES ........................................................................................................................................... 19 
7 
STATISTICAL PLAN ..................................................................................................................................... 20 
7.1 
SAMPLE SIZE DETERMINATION ...................................................................................................................... 20 
7.2 
STATISTICAL METHODS ................................................................................................................................. 20 
7.3 
SUBJECT POPULATION(S) FOR ANALYSIS ....................................................................................................... 20 


 
 
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List of Abbreviations 
 
 
PET 
Positron Emission Tomography 
AD 
Alzheimer’s disease 
MRI 
Magnetic resonance imaging 
18F 
Flourine-18 
MK-6240 
6-Fluoro-3-(1H-pyrrolol[2,3-c]pyridine-1-yl)isoquinolin-5-amine 
Florbetaben 
4-[(E)-2-(4-{2-[2-(2-[18F] fluoroethoxy) ethoxy] ethoxy} phenyl)vinyl]-
N-methylaniline 
CVD 
Cerebrovascular disease 
IV 
intravenous 
Ci 
Curie 
Bq 
Becquerel 
SUV 
Standard uptake value 
SUVR 
Standard uptake value ratio 
Aβ+ 
Amyloid positivity 
FDA 
Federal Drug Administration 
BBB 
Blood-brain barrier 
NFT 
Neurofibrillary tangle 
VT 
Volume of distribution 
M 
Molar 
w/v 
Weight by volume 
v/v 
Volume by volume 
NOAEL 
No-observed-adverse-effect-level 
3H 
Tritium 
Kd 
Equilibrium dissociation constant 
FTD 
Frontotemporal dementia 
ICH 
International Conference on Harmonization 
MNI 
Molecular NeuroImaging 
AE 
Adverse events 
VOI 
Volumes of interest 
BPND 
Binding potential 
HV 
Healthy volunteer 
DVR 
Distribution volume ratio 
SoT 
Standard of Truth 
BSS 
Bielschowsky silver staining 
CERAD 
Consortium to Establish a Registry for Alzheimer’s Disease 
DLB 
Dementia with Lewy bodies 
ECG 
Electrocardiogram 
SAE 
Serious adverse event 
ECI 
Extracorporeal irradiation 
CT 
Computed tomography 
ED 
Effective dose 
Sv 
Sievert 
Gy 
Gray 


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FOV 
Field of view 
MPRAGE 
Magnetization Prepared Rapid Acquisition Gradient Echo 
ROI 
Region of interest 
FS 
FreeSurfer 
CRF 
Case report form 
UP 
Unexpected problem 
HIPAA 
Health Insurance Portability and Accountability Act 
PHI 
Protected health information 


Page 1 
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Study Summary 
Title 
Alzheimer’s PET Imaging in Racially/Ethnically Diverse Adults 
Short Title 
Alzheimer’s PET Imaging in Racially/Ethnically Diverse Adults 
Protocol Number 
AAAR8986 
Phase 
I 
Methodology 
This is a single center PET Study. 
Racially/ethnically diverse subjects with or without a positive family 
history of Alzheimer’s disease (AD) will have one PET scan with 18F-
MK-6240 over a 30 to 60-minute scanning period, and one PET scan 
with 18F-Florbetaben over a 20-minute scanning period. 
Study Duration 
5 years 
Study Center(s) 
Kreitchman PET Center 
Objectives 
The purpose of this study is to examine the extent to which tau 
deposition is related to cognitive function, to determine whether tau 
deposition varies across racial/ethnic groups, to determine the extent to 
which cerebrovascular disease (CVD) is associated with tau pathology, 
and to determine if any of those associations depend on amyloid status. 
Number of Subjects 
150 
Diagnosis and Main 
Inclusion Criteria 
Inclusion criteria: 
•
Aged 35 – 85 years
•
Have either mild cognitive impairment or mild clinical
Alzheimer’s disease; or have no problem with memory or
thinking.
•
Able to participate in all scheduled evaluations and to complete
all required tests and procedures 
•
Considered likely to comply with the study protocol and to have
a high probability of completing the study
Exclusion criteria: 
•
Past or present history of a certain brain disease other than mild
cognitive impairment or mild clinical Alzheimer's disease.
•
Certain significant medical conditions. Examples are
uncontrolled epilepsy or multiple serious injuries.


 
 
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Diagnosis and Main 
Inclusion Criteria 
• Unable to lie still for PET scans. 
• Radiation exposure for research studies in the last year that 
would put you past allowable limits if included in this study. 
• Participation in the last year in a clinical trial for a disease 
modifying drug for AD unless it can be determined that your 
received placebo and not active drug. 
• Conditions that preclude entry into the scanner (e.g. 
claustrophobia, etc.). 
• Inability to have a catheter in your vein for the injection of the 
radioligand (dye). 
• Currently pregnant or breastfeeding. 
Study Product, Dose, 
Route, Regimen 
18F-MK-6240, 5 mCi, intravenous (IV), one injection 
18F-Florbetaben, 8.1 mCi, intravenous (IV), one injection 
Duration of 
administration 
A single dose of 18F-MK-6240, the scan takes approximately 30-60 
minutes. 
A single dose of 18F-Florbetaben, the scan takes approximately 20 
minutes. 
Reference therapy 
N/A 
Statistical 
Methodology 
The primary outcome measure with be regional standardized uptake 
value ratio (SUVR) for 18F-MK-6240 and amyloid positivity (Aβ+) for 
18F-Florbetaben. 
 


 
 
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Introduction 
This document is a protocol for a human research study. This study is to be conducted according 
to US and international standards of Good Clinical Practice (FDA Title 21 part 312 and 
International Conference on Harmonization (ICH) guidelines), applicable government 
regulations and Institutional research policies and procedures.  
1.1 Background 
Alzheimer’s disease (AD) is one of the most serious public health crises in modern times that 
disproportionately affects ethnic and racial minorities[1, 2].  There are currently no effective 
disease-modifying treatments or preventative strategies.  Single pathway pathogenic models 
emphasize a temporal series of biological changes that drives the neurodegeneration associated 
with the AD clinical syndrome.  The formation of amyloid plaques is thought to initiate the 
disease, which stimulates intracellular tau pathology, neurodegeneration, and, ultimately, 
cognitive and functional decline[3, 4].  However, it is now clear that tau deposition often occurs 
prior to or independent of the formation of amyloid pathology[5-9] and an emerging literature 
suggests that other factors, including cerebrovascular disease, may potentiate tau pathology[10-
12].  After several unsuccessful trials that focus on clearance or prevention of amyloid deposition 
to treat AD[13, 14], attention is turning towards tau as a more viable treatment target[15], 
partially due to its tighter link with the clinical symptoms of the disease[16].  It is also becoming 
clearer that treatment and preventative strategies for AD will be most effective during midlife, 
before the onset of frank symptomatology[17], consistent with recent mandates to study AD risk 
factors across the adult lifespan[18].  However, there is very little available information on AD 
biomarkers during midlife and there is an absolute dearth of existing data on biomarker profiles, 
their cognitive correlates, and their risk factors among middle-aged ethnic and racial minorities, 
who are at greater risk for the disease[1, 19].   
1.2 Investigational Agent 
18F-MK-6240 (6-Fluoro-3-(1H-pyrrolo[2,3-c]pyridin-1-yl)isoquinolin-5-amine, 18F-MNI-946) is 
a PET radioligand that binds to paired helical filament tau aggregates. This radioligand has only 
been used for research purposes. 18F-MK-6240 will be administered in tracer doses at activity of 
up to 5 mCi (185 MBq) per injection. 
 
18F-Florbetaben (4-[(E)-2-(4-{2-[2-(2-[
18F] fluoroethoxy) ethoxy] ethoxy} phenyl)vinyl]-N-
methylaniline) is approved and indicated for PET imaging of the brain to estimate β-amyloid 
neuritic plaque density in adult patients with cognitive impairment who are being evaluated for 
AD and other causes of cognitive decline. 
1.3 Preclinical Data 
PET studies in rhesus monkey show that 18F-MK-6240 rapidly crosses the blood-brain barrier 
(BBB) followed by rapid clearance (Figure 1). Distribution is homogeneous across all brain 
regions, as expected since neurofibrillary tangle (NFT) pathology is not present in monkey brain 
(Figure 2). The lack of elevated 18F-MK-6240 retention in white matter compared to gray matter 
is an important characteristic for a NFT tracer that is not achieved by more lipophilic tracer 


 
 
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compounds. The distribution volume (VT) in brain stabilized after 60 minutes, indicating 
influence from radio-metabolites on brain signal is minimal or negligible. 
 
 
Figure 1.        Time-activity Curves for 18F-MK-6240 in Rhesus Monkey. 
 
Figure 2.        18F-MK-6240 Summed PET Image (60-120 min) Overlaid on MRI in Rhesus 
Monkey. Sagittal view; scale is in standard uptake value. 
 
Self-block PET studies (n=3) with 18F-MK-6240 were performed by injecting a high dose of 
unlabeled MK-6240 prior to radiotracer injection. No significant changes in VT with respect to 
baseline were observed (Figure 3) across different brain regions. Additionally, VT was 
homogeneous across different brain regions, demonstrating the lack of specific 18F-MK-6240 
binding, as expected in the healthy rhesus brain. The corresponding summed PET images from 
these studies are shown in Figure 4.   


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Figure 3.        Comparison of VT in Selected Brain Regions Between Baseline and Self-block 
PET Studies for 18F-MK-6240.   
Figure 4.        Summed 18F-MK-6240 PET Images (60-120 min) Overlaid on MRI in Rhesus 
Monkey. Comparison of the Baseline image (left) with the Self-block image 
(right). Images are representative of results in three different monkeys.   
A study was done to determine the potential toxicity of MK-6240 when administered IV (via 
bolus injection) once daily to rats for 7 or 8 days. 
Crl:WI(Han) rats were assigned to 4 groups of 10 females and 10 males each that received 16.6, 
33.3, or 333 µg/kg/day of MK-6240 (hydrochloride salt) in 10 mM acetate buffer, 10% (w/v) 
Captisol™, 0.675% (w/v) sodium chloride, 10% (v/v) ethanol, pH 4, or vehicle only, at a dose 
volume of 5 mL/kg.  The dosing formulations contained the clinical precursor-related impurity at 
a MK-6240 to impurity ratio of approximately 10:2. Assessment of toxicity was based on 
mortality, clinical observations, body weights, food consumption, ophthalmic examinations, and 
clinical and anatomic pathology evaluations. 
There were no unscheduled deaths, no MK˗6240-related clinical signs, and no changes in body 
weight or food consumption. There were no changes in hematological, coagulation, serum 
biochemical examinations or urinalyses, ophthalmic findings, and organ weight changes. There 
were no gross observations or histomorphologic findings, and no MK-6240-related localized 
effects observed at the injection sites. 


Page 6 
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Histomorphologic changes attributable to the administration of Captisol™ were seen in all 
groups, including control with a comparable incidence and severity (very slight) in the kidneys 
and in the lungs.  They consisted of, in the renal cortex, a fine clear vacuolation of the epithelial 
cells lining the proximal tubules in females and males and, in the lung, an accumulation of 
macrophages in the alveoli with occasionally finely vacuolated cytoplasm in males.  Similar 
histomorphologic changes have been seen in rats treated with Captisol™ at this concentration. 
Therefore, such observations were considered unrelated to the administration of MK-6240.  
Based on these findings, the no-observed-adverse-effect-level (NOAEL) was ≥333 µg/kg/day. 
3H-Florbetaben in vitro binding experiments reveal two binding sites (Kd of 16 nM and 135 nM) 
in frontal cortex homogenates from patients with AD. Binding of 18F-florbetaben to β-amyloid 
plaques in post-mortem brain sections from patients with AD using autoradiography correlates 
with both immunohistochemical and Bielschowsky silver stains. 18F-Florbetaben does not bind to 
tau or α-synuclein in tissue from patients with AD. Neither Florbetaben nor non-radioactive 19F-
florbetaben bind to AT8 positive tau deposits in brain tissue from patients with frontotemporal 
dementia (FTD), using autoradiography and immunohistochemistry, respectively. 
1.4 Clinical Data to Date 
MK-6240 was administered at microdoses in accordance with ICH M3(R2) guidelines for an 
exploratory clinical PET trial. In the three elderly healthy volunteers that were included in the 
ongoing brain imaging study, after 18F-MK-6240 injection, the radiotracer uptake in brain was 
high with a peak SUV of ~5, followed by a quick washout from all brain regions. The 
radioactivity distribution in brain was homogenous across regions including gray and white 
matter as expected for a healthy volunteer with no NFTs (Figure 5). Absolute quantification of 
target density in terms of VT was low, consistent with low potential for non-specific, off-target 
(non-NFT) binding.  
Figure 5.        Average PET Image (60 – 90 min) Overlaid on MRI from a 68 Year Old 
Healthy Male Subject Injected with 155 MBq of 18F-MK-6240.  
The color scale for PET image is shown in terms of standardized uptake value               
(SUV) that is normalized to injected radioactivity and body weight. 
In this Phase 0 imaging trial performed by Molecular NeuroImaging (MNI), 18F-MK-6240 
(referred to as 18F-MNI-946 in the MNI report) was used for evaluation as a PET radioligand for 
imaging tau protein deposition, six subjects (3 AD, 3 healthy volunteers) completed the study. 


 
 
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Each subject received one PET scan with 18F-MK-6240. Prior to 18F-MK-6240 each subject 
underwent a 18F-florbetapir PET screening scan for amyloid deposition. The investigational 
radiopharmaceutical, 18F-MK-6240, and imaging procedures were well tolerated with no 
reported adverse events (AEs) related to the product. 
Dynamic PET scans were acquired on a Siemens ECAT EXACT HR+ over 180 minutes 
following 18F-MK-6240 injection (0-90 and 120-180 minute imaging segments). A brain MRI 
was obtained as part of the screening visit. PET and MRI images were aligned and normalized 
into the Montreal Neurological Institute space, and ATLAS-based (Hammers, N30R83) volumes 
of interest (VOI) were applied to the dynamic PET series. 18F-MK-6240 was evaluated in terms 
of kinetic profile (uptake and clearance), binding potential (BPND) computed noninvasively using 
non-invasive Logan with cerebellar cortex as reference, and target region to cerebellum 
standardized uptake value ratio (SUVR) for both healthy volunteers (HVs) and AD subjects. 
18F-MK-6240 exhibited high initial brain uptake within about 3 min of tracer injection with SUV 
values of 3.5-6.0 followed by fast washout in low binding regions (80-85% at ~60 min post-
injection) and moderate in high binding regions (30-50% decrease at ~60 min post injection), 
and by a slow and steady washout in all regions during ~60-180 min interval. Low concentration 
was found in the cerebellum and white matter. 
 
In AD subjects, 18F-MK-6240 showed elevated retention and SUVR values in regions expected 
to demonstrate tau pathology, with values of up to 4.0, compared to ~0.5-1.5 for the HV subjects. 
The pattern of uptake varied between AD patients and between hemispheres, consistent with 
known heterogeneity of tau pathology in AD. 18F-MK-6240 metabolism was rapid with 7 ± 3% 
(n=5) intact parent remaining at 60 min post injection. 
 
The results of this proof-of-concept human study suggest that 18F-MK-6240 can detect tau 
pathology in AD and has suitable pharmacokinetic properties for quantitative measurements of 
tau pathology burden. 
 
Three controls (age 68, 66, and 59 years) and one patient with AD (age 67 years, Mini Mental 
State Exam score 11) underwent PET imaging with 18F-MK-6240 with arterial sampling for full 
quantification of the PET data. Images were acquired 0-90 min post-injection. Total VT was 
calculated using the two-tissue compartment model with the metabolite-corrected arterial input 
function. VT values were similar (~3.5 – 4.5 mL · cm-3) across brain regions in the control 
subjects, with stable values after 60 min of scan time. In the AD patient, VT values were elevated 
(6 – 9 mL · cm-3) in amygdala, parahippocampal gyrus, and medial temporal cortex regions.  
 
The cerebellum was used as reference region to calculate relative cortical binding with both 
kinetic modeling (Two tissue compartment model; distribution volume ration (DVR)) and 
simplified measurement (SUVR) using 60-90 min scan data. DVR and SUVR values showed 
strong correlation (R2 = 0.96, P < 0.0001), suggesting SUVR can be used to quantify 
neurofibrillary tangle binding.   
Florbetaben was evaluated in three single arm clinical studies (Study A-C) that examined images 
from adults with a range of cognitive function, including some end-of-life patients who had 
agreed to participate in a post-mortem brain donation program. Subjects underwent Florbetaben 
injection and scan, then had images interpreted by independent readers masked to all clinical 


Page 8 
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information. The Standard of Truth (SoT) was based on the histopathologic examination using 
Bielschowsky silver staining (BSS) of six brain regions assessed by a Pathology Consensus 
Panel masked to all clinical information (including PET scan results). Florbetaben PET imaging 
results (negative or positive) corresponded to a histopathology derived plaque score based on the 
Consortium to Establish a Registry for Alzheimer’s Disease (CERAD) criteria using neuritic 
plaque counts (Table 1). For the subject level SoT, if in any of the six regions β-amyloid neuritic 
plaques were more than sparse, the subject was classified as positive; if in none of the regions the 
β-amyloid neuritic plaques were assessed as being more than sparse, the subject was classified as 
negative. 
Study A evaluated Florbetaben PET images from 205 subjects and compared the results to 
postmortem truth standard assessments of brain β-amyloid neuritic plaque density in subjects 
who died during the study. The median age was 79 years (range 48 to 98 years) and 52% of the 
subjects were male. By medical history 137 study participants had AD, 31 had other non-AD 
dementia, 5 had dementia with Lewy Bodies (DLB), and 32 had no clinical evidence of 
dementia. Interpretation of images from 82 autopsied subjects was compared to the subject level 
histopathology SoT. Three readers, after undergoing in-person tutoring, interpreted images using 
a clinically applicable image interpretation methodology (See Attachment C). At autopsy, the 
subject level brain β-amyloid neuritic plaque density category was: frequent (n = 31); moderate 
(n = 21); sparse (n = 17); or none (n = 13). Results from Study A are presented in Table 2 and 
Table 3.  


Page 9 
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In Study B five independent, blinded readers underwent the Electronic Media Training in the 
clinically applicable image interpretation methodology (See Attachment C) and assessed images 
from the same 82 end-of-life subjects who enrolled in Study A. The time interval between the 
Florbetaben scan and death was less than one year for 45 patients, between one and two years for 
23 patients and more than two years for 14 patients. Results from Study B can also be found in 
Table 2 and Table 3. 
Study C evaluated the reliability and reproducibility of the clinically applicable image 
interpretation methodology (see Attachment C) using the Electronic Media Training; 461 images 
from previous clinical studies were included from subjects with a range of diagnoses. Five new 
readers assessed randomly provided images from subjects with a truth standard (54 subjects who 
underwent an autopsy) and without a truth standard (51 subjects with mild cognitive impairment, 
182 subjects with AD, 35 subjects with other dementias, 5 subjects with Parkinson’s Disease and 
188 HV). Among the 461 subjects, the median age was 72 years (range 22 to 98), 197 were 
females, and 359 were Caucasian. Image reproducibility data for various subject groups in Study 
C are presented in Table 4. Interreader agreement across all 5 readers had a kappa coefficient of 
0.79 (95% CI 0.77, 0.83). The performance characteristics in 54 subjects with SoT were similar 
to those measured in Studies A and B. Additionally, intra-reader reproducibility was assessed 
from 46 images (10%); the percentage of intra- reader agreement for the 5 readers ranged from 
91% to 98%. 


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1.5 Dose Rationale and Risk/Benefits 
Single IV doses of [18F]MK-6240 (up to ~185 MBq [5 mCi], containing ≤ 20 μg MK-6240) were 
generally well tolerated in healthy male and female subjects. In patients administered with the 
former Captisol formulation, (PN-001), 6 subjects (46.2%) reported a total of 3 treatment 
emergent AEs, of which one was considered by the primary investigator to be related to the 
study drug (frontal headache), (Table 5), which resolved spontaneously, and two were 
considered not related to study drug, rather method of administration (vascular access site 
bruising and vascular access site hematoma). 
In the MNI-946 #e0061 study, using the current formulation, a sterile solution of up to 10% (v/v) 
ethanol, and 0.5% sodium ascorbate in saline (0.9% sodium chloride), one subject reported left 
wrist hematoma which was not considered related to study drug. There was no commonly 
recurring AE reported following [18F]MK-6240 injection. All AEs were mild in intensity.  
In the CE-01-JPN-18-01 study, also using the current formulation, two subjects reported adverse 
events prior to study drug administration. There were no adverse events reported post [18F]MK-
6240 dose. 
Data from the meta-analysis of safety report (May 2020) and the two completed studies indicate 
that of 4,230 subjects exposed to [18F]MK-6240 in ongoing trials at the time of the report, only 
0.95% (39 subjects) experienced an AE and only 0.40% (17 subjects) experienced an AE that 
was considered related to injection of [18F]MK-6240. One subject experienced two related AEs 
(headache and arthragia). There were no SAEs reported that were considered related to injection 
of [18F]MK-6240. One unrelated SAE of pneumonia is recorded in the safety database. Other 
unrelated AEs reported in the meta-safety analysis study population included claustrophobia, 
anxiety, back pain, chest pain, musculoskeletal pain, nausea, hypertension, decreased blood 
glucose, skin irritation, falling and discomfort in the PET scanner. 


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Combined adverse event data from the above and ongoing studies evaluated with the data cutoff 
of 08 May 2022 are presented below. The most commonly occurring AEs that were related to 
injection of [18F]MK-6240 included headache (ten subjects) and dizziness (three subjects). All 23 
of the AEs reported in 22 subjects are listed in Table 5 and were considered possibly related to 
injection of [18F]MK-6240. 
No Suspected Unexpected Serious Adverse Reactions (SUSARs) have been reported (9825 
patients total in completed (n=25) and clinical trials contributing to the global safety database 
(n=9800)).  
There were no commonly recurring AEs reported following treatment with [18F]MK-6240. All 
AEs deemed at least possibly related to [18F]MK-6240 were mild in intensity. 
No clinically significant abnormalities were noted in physical examinations, laboratory safety 
tests (chemistry, hematology, and urinalysis), VS, and ECGs. A meta-analysis of the safety 
database indicated that a majority of the studies contributing BP data demonstrated modest 
decreases in mean systolic and diastolic BP at the Post-injection Pre-scan time point while a 
majority of the studies demonstrated increases in mean systolic and diastolic BP at the Post-
injection Post-scan time point. It is difficult to determine whether these BP changes are related 
to injection with [18F]MK-6240 or whether they are related to the imaging procedures or other 
factors. BP monitoring before administration of an imaging agent and after imaging is routine 
in clinical practice and should continue a best practice for individuals [18F]MK-6240. 
Furthermore, consistent with microdose patient exposures, injection with [18F]MK-6240 does not 
appear to alter ECG recordings in any clinically significant manner. 
There were no ECI, deaths or discontinuations due to an AE. 


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Table 5: Subjects With Adverse Events Considered Related to Study Drug By The 
Investigator (Incidence > 0% in One or More Treatment Groups) 
Completed Studies (n=25) 
MNI-946 #e0061 (n=6) 
PN-001 (n=13) 
CE-01-JPN-18-01 (n=6) 
All studies contributing to the 
global safety database   
(n=9825)a
n
(%)
n
(%)
  Subjects
  with one or more related adverse events 
1 
(4.0) 
22 
(0.22)
Gastrointestinal disorders 
0 
(0.0) 
1 
(0.01) 
    Abdominal distension 
0 
(0.0) 
1 
(0.01) 
General disorders and administrative  
  site conditions 
0 
(0.0) 
2 
(0.02) 
  Injection Site Bruising 
0 
(0.0) 
1 
(0.01) 
  Injection Site Pruritis 
0 
(0.0) 
1 
(0.01) 
 Musculoskeletal and connective tissue 
 disorders 
0 
(0.0) 
1 
(0.01) 
   Arthralgia 
0 
(0.0) 
1b 
(0.01) 
Nervous system disorders 
1
(4.0)
17
(0.17)
     Dizziness 
0 
(0.0) 
4 
(0.04) 
     Dysgeusia 
0 
(0.0) 
1 
(0.01) 
     Headache
1
(4.0)
10
(0.10)
     Parosmia 
0 
(0.0) 
1 
(0.01) 
     Parosmia 
0 
(0.0) 
1 
(0.01) 
Psychiatric disorders 
0 
(0.0) 
1 
(0.01) 
     Insomnia
0
(0.0)
1
(0.01)
Renal and Urinary Disorders 
0 
(0.0) 
1 
(0.01) 
 Dysuria 
0 
(0.0) 
1 
(0.01) 
Every subject is counted a single time for each applicable row and column. 
A system organ class or specific adverse event appears on this report only if its incidence in one or more of the columns meet s 
the incidence criterion in the report title, after rounding. 
a As of the cutoff date 08 May 2022, 9825 subjects had been exposed to [18F]MK-6240 in completed and ongoing trials. This 
data includes expedited reports, if any in ongoing studies. 
b This subject also reported headache and was the only subject in the safety database to report two related adverse events. 
Three (3) healthy volunteers were included in the dosimetry study (two females and one male). 
For each subject, 10 whole body images over approximately 5 hours (18F radionuclide, physical 
half-life is 109.77 min) were serially acquired according to standard procedures on a Siemens 
Biograph PET-CT camera. Three-dimensional volumes of interest were drawn to estimate total 
undecay corrected radioactivity exposures as a function of time in each organ of interest that 
takes up the tracer in significant and visually assessable amounts in addition to others that were 
delineated in the registered CT image (e.g. lungs and muscle). From these radioactivity exposure 
curves, the mean residence time of the radiotracer were calculated for each organ/tissue. These 


 
 
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values were entered into a human biodistribution model (Olinda/EXM) to calculate the effective 
dose (ED).  
 
Evaluation of radiation dosimetry has been performed following administration of  
[18F]MK-6240 in three healthy humans (two females and one male). The average (± SD) value of 
ED was 29.4 ± 0.6 µSv/MBq, which is in the typical range for 18F-radiolabelled ligands. The 
radioactivity widely distributed to various body organs with organ absorbed doses largest for the 
gallbladder (202 µGy/MBq), small intestine (116 µGy/MBq), upper large intestine (128 
µGy/MBq) and urinary bladder (128 µGy/MBq). Based on this, the administration of one 185 
MBq (5 mCi) of [18F]MK-6240 for PET scanning is anticipated to result in a total human ED of 
about 5.4 mSv. 
 
The overall safety profile of Florbetaben is based on data from 978 administrations of 
Florbetaben to 872 subjects and 12 subjects who received vehicle only. No serious adverse 
reactions related to Florbetaben administration have been reported. The most frequently observed 
adverse drug reactions in subjects receiving Florbetaben were injection site reactions consisting 
of erythema, irritation and pain. All adverse reactions were mild to moderate in severity and of 
short duration. 
 
The effective dose resulting from a 300 MBq (8.1 mCi) administration of Florbetaben in adult 
subjects is 5.8 mSv. The use of a CT scan to calculate attenuation correction for reconstruction of 
Florbetaben images (as done in PET/CT imaging) will add radiation exposure. Diagnostic head 
CT scans using helical scanners administer an average of 2.2 ± 1.3 mSv effective dose (CRCPD 
Publication E-07-2, 2007). A low-dose head CT attenuation scan administers a 0.04 mSv 
effective dose. The actual radiation dose is operator and scanner dependent.  
 
Thus, the total combined radiation exposure from MK-6240 and Florbetaben administration and 
subsequent scan on a PET/CT scanner is estimated to be 5.4mSv + 5.8mSv + 2 * 0.04mSv = 
11.28 mSv. 
2 Study Objectives 
The study employs tau PET imaging in a well-characterized multi-racial/ethnic cohort to 
examine the extent to which tau pathology is associated with cognition, differences in tau 
pathology across racial/ethnic groups, and the relationship between MRI markers of small-vessel 
cerebrovascular disease and tau pathology. The study also investigates amyloid-dependent tau 
spreading. 
 
Regional SUVR for 18F-MK-6240 will be calculated to investigate associations with measures of 
memory, olfactory function, and cerebrovascular disease, and amyloid positivity for 18F-
Florbetaben will be calculated to investigate the potential moderation of amyloid on the 
associations with tau. 


 
 
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Figure 6.        Procedures for PET quantification  
  
 
3 Study Design 
3.1 General Design 
Tau and Aβ PET scanning takes place at the Kreitchman PET Center in the Radiology 
Department at Columbia University on separate days within 24 months of each other.  18F-MK-
6240 will be provided by Cerveau and 18F-Florbetaben is provided by Life Molecular Imaging.  
Participant preparation includes intravenous catheterization followed by administration of 8.1 
mCi as a slow single intravenous bolus (6 sec/mL) in a total volume of up to 10 mL of 18F-
Florbetaben for Aβ PET imaging and 5 mCi of 18F-MK-6240 for tau PET.  The 18F- Florbetaben 
PET scans are acquired over a period of 20 minutes in 4 × 5 minute frames on an MCT PET/CT 
scanner (Siemens) in dynamic, 3D imaging mode beginning 90 minutes after injection. The 18F-
MK-6240 PET scans are acquired over a period of 30 to 60 minutes in 6 x 5 to 12 x 5 minute 
frames on the same scanner, beginning 60 to 80 minutes after injection. 
Transmission scans are done prior to the 
emission scan.  An accompanying 
structural CT scan (in-plane 
resolution=0.58×0.58 mm, slice 
thickness=3mm, FOV=29.6×29.6 cm, 
number of slices=75) is also acquired.  For 
quantitative analysis of tau and Aβ PET 
scans[20], dynamic PET frames are 
aligned to the first frame with rigid-body 
registration and a static PET image is 
obtained by averaging the four frames. The 
static PET image is then registered to the 
CT to obtain the inverse transformation 
matrix that transfers the CT image to the 
static PET image space.  Each individual’s 
MPRAGE MRI is also registered to the 
participant’s CT image using normalized mutual information and tri-linear interpolation.  Sets of 
regions-of-interest (ROIs) are selected from the FreeSurfer (FS)-segmented MRI scans.  A 
combination of the two transformation matrices is used to transfer the ROI masks and the 
cerebellar grey matter from MRI space to static PET image space using nearest neighbor 
interpolation. These regional masks are used to extract the regional PET data.  The SUV, defined 
as the decay-corrected brain radioactivity concentration normalized for injected dose and body 
weight, is calculated in selected regions.  The SUV is then normalized to cerebellum grey matter 
to derive the SUVR.  This process is illustrated in Figure 6.       
For tau PET, in asymptomatic individuals tau pathology is typically restricted primarily to the 
medial temporal lobes [21-24] (corresponding to Braak Stage I/II) and tau pathology in this 
region appears to be selectively related to episodic memory [23, 24], so we will use bilaterial 
medial and inferior temporal cortex for the primary analysis of tau imaging.  We will also 
quantitate SUVR in regions corresponding to Braak stage III/IV (limbic cortex), and stage V/VI 
(isocortex) [23] for secondary analyses.  Tau SUVR data in all FreeSurfer-defined cortical 
regions will be databased for exploratory analyses.  
For Aβ PET, SUVR values are derived in lateral temporal cortex, parietal cortex, and posterior 


 
 
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cingulate/precuneus as the primary ROIs.  Using K-means clustering of log-transformed SUVR 
values, we derive cut scores for positivity following established procedures[25].  The overall 
rating is Aβ+ if any of the regions is considered positive.  For the current study we are primarily 
concerned with whether participants are Aβ+, but we will also database continuous amyloid 
SUVR values in all FS-defined ROIs.   
3.2 Primary Study Endpoints 
The primary outcome measure with be regional SUVR values for 18F-MK-6240 and Aβ+ for 18F-
Florbetaben. 
3.3 Primary Safety Endpoints 
The PET/CT imaging in this study is being undertaken for research purposes only and will not be 
reviewed clinically unless specifically requested. The majority of participants will receive or 
have received an MRI scan under a different protocol, which is reviewed clinically. A small 
number of participants are not eligible to receive an MRI scan due to contraindications, and 
therefore will not have had a clinical review. 
 
4 Subject Selection and Withdrawal 
4.1 Inclusion Criteria 
We will limit our recruitment into this study to people who are between 35 and 85 years old at 
the time of enrollment, with or without confirmed WHICAP parent history of AD, and who are 
currently enrolled in the ongoing Offspring Study. Recruitment will focus on those individuals 
who did or will undergo structural MRI scanning as part of this Offspring study within 18 
months of PET study participation. A small number of subjects who enroll in the PET study 
may have contraindications to MRI scanning, for which we have an adjusted method of image 
analysis in place as described in Section 4.3 below.  
The participants must meet the following: 
1. Age 35-85 years  
2. Have either mild cognitive impairment or mild clinical Alzheimer's disease; or have no 
problem with memory or thinking 
3. Able to participate in all scheduled evaluations and to complete all required tests and 
procedures 
4. Must be considered likely to comply with the study protocol and to have a high 
probability of completing the study 
 
4.2 Exclusion Criteria 
Exclusion criteria include pregnant or lactating women, conditions precluding entry into the 
scanners (e.g. claustrophobia, etc.), and inability to have a venous catheter for the injection of the 
radioligand.  
 


 
 
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TABLE 6. Recruitment 
Participants will be excluded from the study if any of the following applies: 
1. Past or present history of certain brain disease other than mild cognitive impairment or 
mild clinical Alzheimer's disease 
2. Certain significant medical conditions. Examples are uncontrolled epilepsy or multiple 
serious injuries 
3. Unable to lie still for the duration of the PET scans 
4. Radiation exposure for research studies in the last year that would put the subject past 
allowable limits if included in this study 
5. Participation in the last year in a clinical trial for a disease modifying drug for AD 
unless it can be determined that the subject received a placebo and not an active drug 
6. Conditions that preclude entry into the scanner (e.g., claustrophobia, etc.). 
7. Inability to have a catheter placed in the vein for the injection of the dye 
8. Currently pregnant or breastfeeding 
 
4.3 Subject Recruitment and Screening 
Adult children from the WHICAP study are currently being recruited into the Offspring Study.  
The parents are a selected subset of participants in WHICAP who are active (seen at the most 
recent assessment wave) or inactive (deceased, moved, too ill to participate) and seen at baseline 
and at least one follow-up.  We recruit from families in which parents with and without AD are 
demographically matched. Table 6 shows the number of WHICAP families from which we are 
recruiting offspring, given the current prevalence of AD among WHICAP participants and the 
expected conversion rate over 5 years among active participants.  Our preliminary data suggest 
that on average, we will have access to two to three offspring who live in the greater NYC area.   
 
We plan to assess 600 offspring per year for all five years of the Offspring Study, and obtain 
medical, neuropsychological, and functional data from an in-person visit. We are recruiting 
approximately 33% of the offspring to participate in brain MRI. These 1,000 (n=200 per year) 
offspring will be representative of the 
cohort at large with respect to parent 
diagnosis, race/ethnicity, and sex.  All 
offspring will be screened for 
eligibility for structural brain MRI. 
Table 6 shows our expected MRI 
recruitment based on our experience in 
the WHICAP study, our pilot interview 
data of offspring, and conservative 
estimates of eligibility and willingness 
to be scanned. Only one offspring per family will be recruited for the MRI.  If there are more than 
one offspring who are eligible for MRI within a family, we select the oldest.  Of eligible families, 
we expect that 75% of offspring will be eligible after screening for MRI contraindications, and of 
those, 60% will be willing to participate (willingness rate was 79% in our survey), and thus we will 
have 1,079 eligible and willing MRI participants.   
White 
Black 
Hispanic 
no 
AD 
AD 
no AD 
AD 
no AD 
AD 
Active WHICAP parents 
547 
100 
494 
206 
903 
505 
Inactive WHICAP parents 
460 
59 
452 
109 
421 
162 
Parents with children (75%) 
755 
119 
710 
236 
993 
500 
Assent/locate (95%/50%) 
562 
93 
521 
188 
801 
421 
Other parent AD-free (90%) 
506 
n/a 
469 
n/a 
721 
n/a 
# offspring avail (2 per fam) 
1012 
187 
939 
375 
1442 
841 
MRI elig. (1 per fam; 75%) 
380 
70 
352 
141 
541 
315 
MRI willing (60%) 
228 
42 
211 
84 
325 
189 


 
 
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Aβ and tau PET scans will be obtained on a representative subset of 150 offspring (with and 
without positive family history of AD) between the age of 35 and 85. We will prioritize 
individuals who have already undergone or will undergo structural MRI as part of the Offspring 
Study. We will obtain both PET scans within 24 months of each other. Therefore, about 33 
individuals per year will be recruited and scanned for contemporaneous Aβ and tau PET and 
MRI scanning.  MRI scans will be completed up to 18 months prior to the first PET scan, or up 
to 18 months after the last PET scan. Since MRI scanning has already commenced in the overall 
study, we will re-acquire MRI scans (with other funds) in instances when the initial scan falls 
outside of the 18-month window. A subset of PET eligible subjects may be ineligible for MRI 
scans due to contraindications. To allow us to continuously meet recruitment goals, we will still 
obtain PET scans on these individuals and use an adjusted statistical method during data 
analysis. Specially, PET data collected without MRI data will be quantified as parametric 
standard uptake value ratio (SUVR) images using atlas-based regions of interest (ROIs) rather 
than subject-specific, MRI-based ROIs. The Montreal Neurological Institute (MNI) brain atlas 
will undergo a non-linear spatial transform to PET space. An inferior cerebellar gray matter ROI 
from the MNI template will be used as reference region. Alzheimer's disease specific ROIs from 
the MNI template will be used to derive mean SUVRs. A covariate for analysis method (atlas-
based ROIs vs subject-specific, MRI-based ROIs) will be included in all statistical analyses.  
 
4.4 Early Withdrawal of Subjects 
4.4.1 When and How to Withdraw Subjects 
Subjects may be withdrawn if they experience any serious adverse reactions to the radioligand or 
during the participation in the study. 
4.4.2 Data Collection and Follow-up for Withdrawn Subjects 
No data collection will occur after withdrawal in subjects who drop out of the study. If data was 
collected prior to withdrawal, such data will be analyzed if possible. 
5 Study Drug  
5.1 Description 
18F-MK-6240 is a novel tau radioligand recently developed by Merck & Co., Inc [26]. 18F-MK-
6240 shows favorable binding properties in both human brain homogenates and rhesus monkey 
brain PET studies [27]. Pharmacological studies have shown that the unlabeled compound MK-
6240 is not expected to have any significant binding to other targets that would confound 
measurement of tau aggregation. Early human PET studies have shown that 18F-MK-6240 
binding is elevated in AD patients, and that resulting radiation exposure is within the range of 
other 18F radioligands. 
 
Florbetaben is indicated for PET imaging of the brain to estimate β-amyloid neuritic plaque 
density in adult patients with cognitive impairment who are being evaluated for AD and other 
causes of cognitive decline. A negative Florbetaben scan indicates sparse to no amyloid neuritic 
plaques and is inconsistent with a neuropathological diagnosis of AD at the time of image 
acquisition; a negative scan result reduces the likelihood that a patient’s cognitive impairment is 


 
 
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due to AD. A positive Florbetaben scan indicates moderate to frequent amyloid neuritic plaques; 
neuropathological examination has shown this amount of amyloid neuritic plaque is present in 
patients with AD but may also be present in patients with other types of neurologic conditions as 
well as older people with normal cognition. Florbetaben is an adjunct to other diagnostic 
evaluations. 
 
5.2 Preparation and Administration of Study Drug 
Participant preparation includes intravenous catheterization followed by administration of 8.1 
mCi as a slow single intravenous bolus (6 sec/mL) in a total volume of up to 10 mL of 18F-
Florbetaben for Aβ PET imaging and 5 mCi of 18F-MK-6240 for tau PET. 
 
18F-MK-6240 will be formulated in clear solution of up to 10% (v/v) ethanol, and 0.5% sodium 
ascorbate in saline (0.9% sodium chloride) for IV bolus administration.  
 
18F-Florbetaben is supplied in a 30 mL glass vial containing up to 30 mL of a clear solution at a 
strength of 50 to 5000 MBq/mL (1.4 to 135 mCi/mL) 18F-florbetaben at end of synthesis. Each 
vial contains multiple doses and is enclosed in a shielded container to minimize external 
radiation exposure. 
5.3 Packaging 
N/A since 18F-MK-6240 will be packaged and provided by Cerveau and 18F-Florbetaben will be 
packaged and provided by Life Molecular Imaging. 
5.4 Receiving, Storage, Dispensing and Return 
5.4.1 Receipt of Drug Supplies 
Upon receipt of the of the study treatment supplies, an inventory must be performed and a 
drug receipt log filled out and signed by the person accepting the shipment. It is 
important that the designated study staff counts and verifies that the shipment contains all 
the items noted in the shipment inventory.  Any damaged or unusable study drug in a 
given shipment (active drug or comparator) will be documented in the study files.  The 
investigator must notify study sponsor of any damaged or unusable study treatments that 
were supplied to the investigator’s site. 
 
5.4.2 Storage 
Store 18F-Florbetaben and 18F-MK-6240 at room temperature 25°C (77°F); excursions 
permitted to 2°C to 42°C (36°F to 108°F). The product does not contain a preservative. 
Store within the original container or equivalent radiation shielding. Radiotracer must not 
be diluted. 
 
This preparation is approved for use by persons under license by the Nuclear Regulatory 
Commission or the relevant regulatory authority of an Agreement State. 


 
 
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5.4.3 Dispensing of Study Drug 
Slow bolus (6 sec/mL) IV injection of 5 mCi 18F-MK-6240 and 8.1 mCi 18F-Florbetaben 
on the same day as delivery to the Columbia University Medical Center PET Department. 
 
For 18F-MK-6240, the carrier mass dose will be limited to no more than 20 μg per 
injection. For 18F-Florbetaben, the carrier mass dose will be limited to no more than 30 
μg per injection. 
5.4.4 Return or Destruction of Study Drug 
At the completion of the study, there will be a final reconciliation of drug shipped, drug 
consumed, and drug remaining.  This reconciliation will be logged on the drug 
reconciliation form, signed and dated.  Any discrepancies noted will be investigated, 
resolved, and documented prior to return or destruction of unused study drug.  Drug 
destroyed on site will be documented in the study files. 
6 Study Procedures 
6.1 Tau PET 
The 18F-MK-6240 PET scans are acquired over a period between 30 to 60 minutes in 6 to 12 x 
5  minute frames on the same scanner, beginning 60 to 80 minutes after injection. 
 
Transmission scans are done prior to the emission scan.  An accompanying structural CT scan 
(in-plane resolution=0.58×0.58 mm, slice thickness=3mm, FOV=29.6×29.6 cm, number of 
slice=75) is also acquired. 
 
6.2 Amyloid PET 
The 18F-Florbetaben PET scans are acquired over a period of 20 minutes in 4 × 5 minute frames 
on an MCT PET/CT scanner (Siemens) in dynamic, 3D imaging mode beginning 90 minutes 
after injection. 
 
Transmission scans are done prior to the emission scan.  An accompanying structural CT scan 
(in-plane resolution=0.58×0.58 mm, slice thickness=3mm, FOV=29.6×29.6 cm, number of 
slice=75) is also acquired. 
 
The second PET visit will occur within 24 months of the first. There is no particular order that 
the participants must receive the study drugs. If eligible, the MRI scan will be collected up to 18 
months prior to the first PET visit, or up to 18 months after the second PET visit. 
 
6.3 Questionnaires 
Participants may be asked to complete any number of outstanding questionnaires during the PET 
scan uptake periods. The questionnaires are self-administered and are part of the planned data 
collection for the main Offspring study. 
 


 
 
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7 Statistical Plan 
7.1 Sample Size Determination 
The sample size of 150 subjects was selected to allow sufficient power (  80%) of a two-sided 
significance test with  = 0.05 to detect meaningful effects associated with the primary 
hypotheses.  We used the G*Power[28] to perform the following power analyses.   
7.2 Statistical Methods 
Regional SUVR for 18F-MK-6240 will be calculated to investigate associations with measures of 
memory, olfactory function, and cerebrovascular disease, and amyloid positivity for 18F-
Florbetaben will be calculated to investigate the potential moderation of amyloid on the 
associations with tau. 
Before any specific statistical modelling techniques are applied, we will examine all variables for 
illegitimate values, outliers, and other inconsistencies.  Distributions of demographic and other 
clinically important variables will be examined and summarized by means, standard deviations, 
minima, and maxima for continuous measures and proportions for categorical measures.  Similar 
descriptive analyses will be conducted on all outcome variables of interest.  We will make every 
effort to obtain all data to reduce or eliminate missing data issues.  Tests will be two-sided and 
performed at a significance level of =0.05.  After descriptive examination of the variables of 
interest, a series of unadjusted or partially adjusted planned analyses will be conducted to 
address the specific hypotheses, followed by fully adjusted models, follow-up analyses, and 
exploratory analyses.  In all regression analyses, steps will be taken to avoid issues related to 
multicollinearity of the predictors.  If necessary, data reduction techniques (e.g., screening out 
variables that are highly correlated) will be applied to the predictors prior to fitting the regression 
models. 
7.3 Subject Population(s) for Analysis 
The study builds on an over-25-year history of studying cognitive aging and dementia in the 
community through the WHICAP study and the newly-established Offspring Study, in which 
3,000 adult children of participants in the WHICAP study are being recruited and evaluated 
comprehensively, and 150 Offspring participants will undergo PET imaging. This 
racially/ethnically diverse subject population will allow the examination of sources of health 
disparities and the examination of biomarkers in established community-based diverse cohorts, 
utilizing existing cohorts. We have detailed characterization of both the parental and offspring 
generations, including consensus diagnosis based on detailed in-person assessment, longitudinal 
cognitive data, risk factors, and date of onset.  Unlike many other offspring cohorts, our 
Offspring Study has direct assessment of parental AD status among a community-based sample.  
Finally, we will be at the leading edge of examining the interplay of cerebrovascular disease and 
AD pathology, both of which occur at higher prevalence in racial/ethnic minorities, by being 
among the first to examine the relationship between markers of cerebrovascular disease and tau 
pathology in vivo. 


 
 
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14 References 
[1] Tang MX, Cross P, Andrews H, Jacobs DM, Small S, Bell K, et al. Incidence of Alzheimer's 
disease in African-Americans, Caribbean Hispanics and Caucasians in northern Manhattan. 
Neurology. 2001;56:49-56. 
[2] Mayeda ER, Glymour MM, Quesenberry CP, Whitmer RA. Inequalities in dementia 
incidence between six racial and ethnic groups over 14 years. Alzheimers Dement. 2016. 
[3] Jack CR, Jr., Knopman DS, Jagust WJ, Petersen RC, Weiner MW, Aisen PS, et al. Tracking 
pathophysiological processes in Alzheimer's disease: an updated hypothetical model of dynamic 
biomarkers. The Lancet Neurology. 2013;12:207-16. 
[4] Jack CR, Jr., Knopman DS, Jagust WJ, Shaw LM, Aisen PS, Weiner MW, et al. Hypothetical 
model of dynamic biomarkers of the Alzheimer's pathological cascade. The Lancet Neurology. 
2010;9:119-28. 
[5] Braak H, Del Tredici K. The pathological process underlying Alzheimer's disease in 
individuals under thirty. Acta neuropathologica. 2011;121:171-81. 
[6] Crary JF, Trojanowski JQ, Schneider JA, Abisambra JF, Abner EL, Alafuzoff I, et al. 
Primary age-related tauopathy (PART): a common pathology associated with human aging. Acta 
neuropathologica. 2014;128:755-66. 
[7] Jack CR, Jr., Knopman DS, Chetelat G, Dickson D, Fagan AM, Frisoni GB, et al. Suspected 
non-Alzheimer disease pathophysiology--concept and controversy. Nat Rev Neurol. 
2016;12:117-24. 
[8] Duyckaerts C, Braak H, Brion JP, Buee L, Del Tredici K, Goedert M, et al. PART is part of 
Alzheimer disease. Acta neuropathologica. 2015;129:749-56. 
[9] Braak H, Braak E. Frequency of stages of Alzheimer-related lesions in different age 
categories. Neurobiol Aging. 1997;18:351-7. 
[10] Koike MA, Green KN, Blurton-Jones M, Laferla FM. Oligemic hypoperfusion differentially 
affects tau and amyloid-{beta}. The American journal of pathology. 2010;177:300-10. 
[11] Rizvi B, Narkhede A, Last BS, Budge M, Tosto G, Manly JJ, et al. The effect of white 
matter hyperintensities on cognition is mediated by cortical atrophy. Neurobiol Aging. 
2018;64:25-32. 
[12] Zlokovic BV. Neurovascular pathways to neurodegeneration in Alzheimer's disease and 
other disorders. Nat Rev Neurosci. 2011;12:723-38. 
[13] Harrison JR, Owen MJ. Alzheimer's disease: the amyloid hypothesis on trial. The British 
journal of psychiatry : the journal of mental science. 2016;208:1-3. 
[14] Le Couteur DG, Hunter S, Brayne C. Solanezumab and the amyloid hypothesis for 
Alzheimer’s disease. BMJ. 2016;355. 
[15] Holtzman DM, Carrillo MC, Hendrix JA, Bain LJ, Catafau AM, Gault LM, et al. Tau: From 
research to clinical development. Alzheimers Dement. 2016;12:1033-9. 
[16] Nelson PT, Alafuzoff I, Bigio EH, Bouras C, Braak H, Cairns NJ, et al. Correlation of 
Alzheimer disease neuropathologic changes with cognitive status: a review of the literature. 
Journal of neuropathology and experimental neurology. 2012;71:362-81. 
[17] Winblad B, Amouyel P, Andrieu S, Ballard C, Brayne C, Brodaty H, et al. Defeating 
Alzheimer's disease and other dementias: a priority for European science and society. The Lancet 
Neurology. 2016;15:455-532. 
[18] Montine TJ, Koroshetz WJ, Babcock D, Dickson DW, Galpern WR, Glymour MM, et al. 
Recommendations of the Alzheimer's disease-related dementias conference. Neurology. 
2014;83:851-60. 


Page 29 
[19] Graff-Radford NR, Besser LM, Crook JE, Kukull WA, Dickson DW. Neuropathologic
differences by race from the National Alzheimer's Coordinating Center. Alzheimers Dement.
2016;12:669-77.
[20] Gu Y, Razlighi QR, Zahodne LB, Janicki SC, Ichise M, Manly JJ, et al. Brain amyloid
deposition and longitudinal cognitive decline in nondemented older subjects: Results from a
multi-ethnic population. PLoS One. 2015;10:e0123743.
[21] Harada R, Okamura N, Furumoto S, Furukawa K, Ishiki A, Tomita N, et al. 18F-THK5351:
A Novel PET Radiotracer for Imaging Neurofibrillary Pathology in Alzheimer Disease. J Nucl
Med. 2016;57:208-14.
[22] Shah M, Catafau AM. Molecular Imaging Insights into Neurodegeneration: Focus on Tau
PET Radiotracers. J Nucl Med. 2014;55:871-4.
[23] Scholl M, Lockhart SN, Schonhaut DR, O'Neil JP, Janabi M, Ossenkoppele R, et al. PET
Imaging of Tau Deposition in the Aging Human Brain. Neuron. 2016;89:971-82.
[24] Maass A, Lockhart SN, Harrison TM, Bell RK, Mellinger T, Swinnerton K, et al. Entorhinal
tau pathology explains episodic memory and neurodegeneration in normal aging. Manuscript
submitted for publication submitted.
[25] Villemagne VL, Mulligan RS, Pejoska S, Ong K, Jones G, O'Keefe G, et al. Comparison of
11C-PiB and 18F-florbetaben for Abeta imaging in ageing and Alzheimer's disease. European
journal of nuclear medicine and molecular imaging. 2012;39:983-9.
[26] Walji AM, Hostetler ED, Selnick H, Zeng Z, Miller P, Bennacef I, et al. Discovery of 6-
(Fluoro-(18)F)-3-(1H-pyrrolo[2,3-c]pyridin-1-yl)isoquinolin-5-amine ([(18)F]-MK-6240): A
Positron Emission Tomography (PET) Imaging Agent for Quantification of Neurofibrillary
Tangles (NFTs). J Med Chem. 2016;59:4778-89.
[27] Hostetler ED, Walji AM, Zeng Z, Miller P, Bennacef I, Salinas C, et al. Preclinical
Characterization of 18F-MK-6240, a Promising PET Tracer for In Vivo Quantification of
Human Neurofibrillary Tangles. J Nucl Med. 2016;57:1599-606.
[28] Faul F, Erdfelder E, Lang AG, Buchner A. G*Power 3: a flexible statistical power analysis
program for the social, behavioral, and biomedical sciences. Behav Res Methods. 2007;39:175-
91.