Clinical trial NCT02702102
Conditions: Diffuse Lewy Body Disease Dementia With Lewy Bodies Parkinson's Disease Dementia
Sponsor: Columbia University
Start date: 2015-12-01; Completion date: 2019-09-04
Enrollment: 5 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
This study uses a special type of scan called a positron emission tomography (PET) scan to take pictures of the brain. During the PET scan, a special dye called 11C-PBR28 is injected into the body.

11C-PBR28 sticks to parts of the brain where there is inflammation. The purpose of this study is to see if 11C-PBR28 can detect inflammation in patients with Parkinson's disease dementia or dementia with Lewy bodies.

11C-PBR28 is considered a drug by the Food and Drug Administration. 11C-PBR28 is not a treatment for any disease. Rather, 11C-PBR28 can be used to measure inflammation in the brain.

Detailed description:
This study is being done to see if 11C-PBR28 could be useful in diffuse Lewy body disease. Diffuse Lewy Body Disease refers to subjects with either Parkinson's disease dementia or dementia with Lewy bodies.

The hypothesis is that 11C-PBR28 binding will be greater in patients with diffuse Lewy body disease than in controls, with largest differences in temporal and limbic regions of the brain.

Participants will have a medical history and examination. This will include questions about thinking and memory. Participants will have blood drawn for routine lab tests. Blood will also be tested for a special genetic test to make sure the 11C-PBR28 scan will work. A special genetic blood test will also be done to see if participants have genes that are associated with dementia with Lewy bodies or other causes of dementia. Participants will not receive the results of these special genetic tests. Participants will also have a magnetic resonance imaging (MRI) scan of the brain.

If a participant still meet the inclusion and exclusion criteria after these procedures are done, then the participant will have one PET scan using 11C-PBR28. This scan will take place at the Columbia University Medical Center Kreitchman PET Center. On the day of the PET scan you will have an intravenous (IV) catheter (tube) placed in the participant's arm. The participant will also have an arterial catheter placed in an artery of your wrist. Participant will be asked to lay down on a PET scanner. A CT scan will be taken of the participant's head. A small amount of 11C-PBR28 will be injected into the IV. A member of the study team will draw blood from the arterial catheter. The PET scan will last 90 minutes. The participant will be asked to lay still during the scan.

The amount of radiation from the 11C-PBR28 PET scan is up to 20 millicuries. Similar amounts of 11C-PBR28 have been given to over 100 human subjects in prior studies.

Arms: arm 1: Participants will receive one IV injection of up to 20 millicuries of 11C-PBR28.
Interventions: intervention 1: 11C-PBR28
Intervention descriptions: intervention 1: Up to 20 millicuries

Protocol documents:
Prot_SAP_000.pdf: 
 
 
 NCT02702102 
  
  
  
  
  
  
  
IMAGING INFLAMMATION IN PATIENTS WITH DIFFUSE  
LEWY BODY DISEASE  
 
 
Regulatory Sponsor:  
William Charles Kreisl, MD  
Taub Institute  
622 W. 168th St, PH 19, New York, NY 10032 
212-305-9194  
Funding Sponsor:  
Taub Institute  
622 W. 168th St, PH 19, New York, NY 10032  
212-305-9194  
Study Product:  
11C-PBR28  
Protocol Number:  
AAAQ0756  
Date: August 18, 2015  
 
Amended: May 13, 2016  
 
Administrative Change:  
 
 
 
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.  


  
  
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Table of Contents  
STUDY SUMMARY ................................................................................................................................................... 1 
1  INTRODUCTION ................................................................................................................................................... 4 
1.1  BACKGROUND .................................................................................................................................................... 4 
1.2  INVESTIGATIONAL AGENT .................................................................................................................................. 5 
1.3  PRECLINICAL DATA ............................................................................................................................................ 5 
1.4 CLINICAL DATA TO DATE .................................................................................................................................... 9 
1.5 DOSE RATIONALE AND RISK/BENEFITS ............................................................................................................. 11 
2 STUDY OBJECTIVES .......................................................................................................................................... 11 
3 STUDY DESIGN .................................................................................................................................................... 11 
3.1  GENERAL DESIGN ............................................................................................................................................. 11 
3.2  PRIMARY STUDY ENDPOINTS ........................................................................................................................... 13 
3.3 SECONDARY STUDY ENDPOINTS ....................................................................................................................... 13 
3.4 PRIMARY SAFETY ENDPOINTS ........................................................................................................................... 13 
4 SUBJECT SELECTION AND WITHDRAWAL ................................................................................................ 13 
4.1  INCLUSION CRITERIA ........................................................................................................................................ 13 
4.2  EXCLUSION CRITERIA....................................................................................................................................... 14 
4.3 SUBJECT RECRUITMENT AND SCREENING.......................................................................................................... 14 
4.4 EARLY WITHDRAWAL OF SUBJECTS .................................................................................................................. 15 
4.4.1  When and How to Withdraw Subjects ...................................................................................................... 15 
4.4.2 Data Collection and Follow-up for Withdrawn Subjects .......................................................................... 15 
5 STUDY DRUG ........................................................................................................................................................ 15 
5.1  DESCRIPTION .................................................................................................................................................... 15 
5.2 TREATMENT REGIMEN ...................................................................................................................................... 15 
5.3 METHOD FOR ASSIGNING SUBJECTS TO TREATMENT GROUPS .......................................................................... 15 
5.4  PREPARATION AND ADMINISTRATION OF STUDY DRUG ................................................................................... 15 
5.5  SUBJECT COMPLIANCE MONITORING ............................................................................................................... 15 
5.6  PRIOR AND CONCOMITANT THERAPY ............................................................................................................... 15 
5.7  PACKAGING ...................................................................................................................................................... 15 
5.8 BLINDING OF STUDY DRUG ............................................................................................................................... 16 
5.9 RECEIVING, STORAGE, DISPENSING AND RETURN ............................................................................................. 16 
5.9.1  Receipt of Drug Supplies ......................................................................................................................... 16 
5.9.2  Storage ..................................................................................................................................................... 16 
5.9.3  Dispensing of Study Drug ........................................................................................................................ 16 
5.9.4  Return or Destruction of Study Drug ....................................................................................................... 16 
6 STUDY PROCEDURES ........................................................................................................................................ 16 
7 STATISTICAL PLAN ........................................................................................................................................... 18 
7.1  SAMPLE SIZE DETERMINATION ........................................................................................................................ 18 
7.2 STATISTICAL METHODS ..................................................................................................................................... 18 
7.3 SUBJECT POPULATION(S) FOR ANALYSIS .......................................................................................................... 18 
8 SAFETY AND ADVERSE EVENTS .................................................................................................................... 18 
8.1  DEFINITIONS ..................................................................................................................................................... 18 
8.2 RECORDING OF ADVERSE EVENTS ..................................................................................................................... 20 
8.3 REPORTING OF SERIOUS ADVERSE EVENTS ....................................................................................................... 21 
8.3.1 IRB Notification by Investigator ............................................................................................................... 21 
8.3.2 FDA Notification by Investigator .............................................................................................................. 22 
8.4 UNBLINDING PROCEDURES ................................................................................................................................ 22 
8.5 STOPPING RULES ............................................................................................................................................... 22 
8.6  MEDICAL MONITORING .................................................................................................................................... 23 
9  DATA HANDLING AND RECORD KEEPING ................................................................................................ 23 
9.1  CONFIDENTIALITY ............................................................................................................................................ 23 
9.2  SOURCE DOCUMENTS ....................................................................................................................................... 23 
9.3  CASE REPORT FORMS ....................................................................................................................................... 23 


 
  
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10  STUDY MONITORING, AUDITING, AND INSPECTING .......................................................................... 24 
10.1  STUDY MONITORING PLAN ............................................................................................................................ 24 
11 ETHICAL CONSIDERATIONS ........................................................................................................................ 24 
12 STUDY FINANCES ............................................................................................................................................. 24 
12.1  FUNDING SOURCE .......................................................................................................................................... 24 
12.2 CONFLICT OF INTEREST ................................................................................................................................... 24 
12.3 SUBJECT STIPENDS OR PAYMENTS................................................................................................................... 25 
13  PUBLICATION PLAN ....................................................................................................................................... 25 
14  REFERENCES .................................................................................................................................................... 25 
 
  
  
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List of Abbreviations  
  
  
  
PET  
Positron emission tomography  
11C-PBR28  
11C-[O-methyl-11C-N-acetyl-N-(2-methoxybenzyl)-2-phenoxy-
5pyridinamine (a radioligand for inflammation)  
MRI  
Magnetic resonance imaging  
DLBD  
Diffuse Lewy Body Disease  
AD  
Alzheimer’s disease  
PD  
Parkinson’s disease  
DLB  
Dementia with Lewy bodies  
EKG  
Electrocardiogram  
TSPO  
18 kDa translocator protein  
SNP  
Single nucleotide polymorphism  
MCI  
Mild cognitive impairment  
SUVR  
Standardized uptake value ratio  
HAB  
High affinity binder  
MAB  
Mixed affinity binder  
LAB  
Low affinity binder  


 
 
Clinical Research Protocol Template  
  
page 1  
Version 4  
Study Summary  
Title  
Imaging inflammation in patients with diffuse Lewy body disease  
Short Title  
Imaging inflammation in DLBD  
Protocol Number  
AAAQ0756  
Phase  
II  
Methodology  
This is a single center PET study.  
  
Screening will include history, physical examination, routine laboratory studies, 
neurological examination, genetic testing for 18 kDa translocator protein (TSPO) 
status, and brain MRI. Clinical evalution will include Montreal Cognitive 
Assessment (MoCA) and Unified Parkinson’s disease rating scale (UPDRS). 
Subjects determined to have dementia with Lewy bodies or Parkinson’s disease 
dementia (collectively considered as diffuse Lewy body disease) or cognitively 
normal by consensus diagnosis will be included. Subjects will then have one PET 
scan with 11C-PBR28. Subjects who have no previously had genetic testing for 
glucocerebrosidase (GBA) or ApoE will have these tests performed.  
Study Duration  
1 year  
Study Center(s)  
Taub Institute, CUMC   
Objectives  
The primary objective is to generate pilot data in patients with DLBD. The aims are:  
1. 
Determine feasibility of performing 11C-PBR28 PET in patients with DLBD.  
2. 
Determine effect size of 11C-PBR28 binding in patients with DLBD vs. 
controls. The exploratory aims are:  
1. Identify potential reference regions for 11C-PBR28 binding in patients with DLBD. 
2. Determine if 11C-PBR28 binding correlates with cognitive impairment and 
Parkinsonism in patients with DLBD.  
Number of Subjects  
Target enrollment is 8 patients and 8 controls. 
Target accrual is 4 patients and 4 controls.  


 
  
 
 
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Diagnosis and Main 
Inclusion Criteria  
Inclusion criteria for patients:  
1. Age 60 and older  
2. Meet criteria for either a) dementia with Lewy bodies, or b) Parkinson’s 
disease dementia  
3. Able to provide informed consent   
4. Written and oral fluency in English   
5. Able to participate in all scheduled evaluations and to complete all required 
tests and procedures.  
6. In the opinion of the investigator, the subject must be considered likely to 
comply with the study protocol and to have a high probability of completing 
the study.  
  
Inclusion criteria for controls:  
1. Age 60 and older  
2. Normal cognitive and motor function based on neurological examination  
3. Written and oral fluency in English   
4. Able to participate in all scheduled evaluations and to complete all required 
tests and procedures.  
5. In the opinion of the investigator, the subject must be considered likely to 
comply with the study protocol and to have a high probability of completing 
the study.  
  
Exclusion criteria:  
1. Past or present history of certain brain disorders (other than DLB or PDD for 
patient participants).  
2. Certain significant medical conditions, which make study procedures of the 
current study unsafe. Such serious medical conditions include uncontrolled 
epilepsy and multiple serious injuries.   
3. Contraindication to MRI scanning  
4. Conditions precluding entry into the scanners (e.g. morbid obesity, 
claustrophobia, etc.).  
5. Exposure to research related radiation in the past year that, when combined 
with this study, would place subjects above the allowable limits.  
6. Low affinity binding on TSPO genetic screen  
7. Currently taking anticoagulant drugs (e.g., warfarin).  
8. Women who are able to become pregnant.  
  
  
Study Product, Dose, 
Route, Regimen  
11C-PBR28, up to 20 mCi (740 MBq), IV, total of one injection.  
Duration of 
administration  
A single dose of radioligand will be injected over 1 minute for the PET scan.   


 
  
 
 
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Reference therapy  
N/A  
Statistical  
Methodology  
Primary outcome  
PET images will be analyzed using 1) the two-tissue kinetic model to calculate total 
distribution volume, corrected for free fraction of radioligand in plasma (VT/fP), and 2) 
the cerebellum as a pseudo-reference region to calculate standardized uptake value 
ratios (SUVR). Effect size will be calculated for different regions as the difference in 
means (DLBD vs. controls) divided by the standard deviation.  
  
Secondary outcomes  
Coefficient of variation (%COV = standard deviation / mean) will be calculated for 
controls and patients with DLBD for both VT/fP and SUVR data. SUVR values will be 
calculated using cerebellum as reference regions. Additional reference regions will be 
explored. Correlation coefficients will be computed to look for correlation between 
11C-PBR28 binding (VT/fP and SUVR) and clinical severity. Severity will be 
measured using CDR-sum of boxes, MoCA score, and UPDRS. We will also look for 
correlations between 11C-PBR28 binding and atrophy, as determined using ROI 
volumes derived from MRI data.  
  
Sample size determination  
This is a Phase 2 pilot study to determine feasibility and preliminary data for a larger 
study. Therefore, the proposed sample size is not expected to provide enough 
statistical power to see significant group differences.  
  
 


 
  
 
 
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1 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 guidelines), applicable government regulations and 
Institutional research policies and procedures.   
1.1 Background  
Neuroinflammation is a proposed pathogenic contributor to diffuse Lewy body disease 
(DLBD), which includes Parkinson’s disease dementia and dementia with Lewy bodies. DLBD 
is defined by aggregated α-synuclein inclusions (i.e., Lewy bodies), which activate microglia in 
vitro and stimulate release of proinflammatory mediators[1]. Activated microglia are found 
proximal to Lewy bodies at autopsy[2-4]. The amount of activated microglia is proportional to 
both the number of Lewy bodies found at autopsy[3] and the amount of α-synuclein and loss of 
dopaminergic neurons measured in transgenic mice[5]. However, the exact role of inflammation 
in the progression of DLBD remains poorly understood. Activated microglia surrounding 
degenerating Lewy body-containing neurons could simply be a response to upstream cellular 
damage[6]. Yet, α-synuclein activates microglia prior to loss of Lewy body-containing 
neurons[7], suggesting an early role of inflammation in pathogenesis. Microglia may play a 
protective role by producing antioxidant enzymes proximal to Lewy body-containing neurons in 
response to oxidative stress[4]. However, microglia produce reactive oxygen species in response 
to α-synuclein[1], and transgenic mouse studies have shown that 1) α-synuclein-induced 
neurodegeneration does not occurs in the absence of chronic inflammation, and 2) inhibiting 
oxidative enzymes produced by microglia ameliorates this neurodegeneration[5]. Therefore, 
dysfunctional glial response to Lewy body pathology may accelerate neuronal loss, and 
attenuating the neuroimmune response to α-synuclein aggregation may have therapeutic benefit.  
A potential complication in studying inflammation in dementia with Lewy bodies is the 
frequent co-existence of Alzheimer’s disease (AD) pathology [8-11], as β-amyloid is also a 
proposed trigger for neuroinflammation[12]. However, autopsy studies have shown microglial 
activation in patients with “pure” dementia with Lewy bodies without AD pathology[3]. 
Glucocerebrosidase (GBA) gene mutations are more prevalent in patients with “pure” dementia 
with Lewy body patients than in those with AD co-pathology [13,14], while patients with 
dementia with Lewy bodies who are ApoE4 non-carriers have lower Braak stage [15,16] and less 
amyloid on PET[17] than ApoE4 carriers. Therefore, selecting DLBD patients with either GBA 
mutations or ApoE4 non-carrier status would reduce the incidence of AD co-pathology and 
facilitate investigation of the relationship between inflammation and synuclein in the absence of 
amyloidopathy.   
Activated microglia over-express the 18 kDa translocator protein (TSPO) in response to 
cellular injury[18], and TSPO density can be quantified in vivo using positron emission 
tomography (PET). Only two previous PET studies have measured TSPO in DLBD. In the first, 
patients with Parkinson’s disease dementia showed greater TSPO binding with 11C-(R)-PK 
11195 than controls in anterior and posterior cingulate, striatum, frontal, temporal, parietal, and 
occipital cortical regions, while non-demented Parkinson’s disease patients showed greater 
binding in temporal, parietal, and occipital regions[19]. In the second, patient’s with dementia 
with Lewy bodies and patients with Parkinson’s disease showed greater binding than controls in 
substantia nigra and putamen, while patients with dementia with Lewy bodies also showed 


 
  
 
 
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greater binding in several associative cortices and cerebellum[20]. While studies have shown 
cerebellar atrophy in dementia with Lewy bodies[21], and some studies have shown α-synuclein 
deposits in cerebellum[22], others have shown decreased[23] or infrequent α-synuclein in this 
region[24]. Therefore, whether TSPO density is increased in cerebellum in living DLBD patients 
has not been conclusively established.  
11C-(R)-PK 11195 has low specific-to-nonspecific binding[25] which could confound 
accurate quantification of TSPO density. The second generation TSPO radioligand 11C-PBR28 
has greater specific-to-nonspecific binding than 11C-(R)-PK 11195 in monkey brain[25] and has 
been used to detect neuroimmune activation in stroke, schizophrenia, and epilepsy[26-28]. 
Therefore, 11C-PBR28 PET imaging is expected to detect inflammatory changes in DLBD and 
may be useful for monitoring disease progression and response to novel treatments. Because the 
relationship between inflammation and the clinical symptoms exhibited in DLDB remains 
unknown, there is a critical need to determine the relationship between inflammation and DLBD 
to increase knowledge about the mechanisms underlying the disease and help develop 
antiinflammatory treatments as disease-modifying therapy for this presently incurable disorder.  
One limitation of 11C-PBR28, shared by all tested second generation TSPO radioligands, is 
differential affinity caused by the rs6971 single nucleotide polymorphism on the TSPO gene[29]. 
Statistical correction for TSPO genotype allows inclusion of individual heterozygous for the 
polymorphism; however, homozygotes have negligible binding with 11C-PBR28 and must be 
excluded from analysis[27].  
The primary research objective is to generate pilot data in patients with DLBD to more 
effectively design an R01 application to comprehensively study the relationship between 
inflammation and DLBD. The aims are to:  
1. Determine feasibility of performing 11C-PBR28 PET in patients with DLBD.  
2. Determine effect size of 11C-PBR28 binding in patients with DLBD vs. controls. The 
exploratory aims are:  
1. 
Identify potential reference regions for 11C-PBR28 binding in patients with 
DLBD.  
2. 
Determine if 11C-PBR28 binding correlates with cognitive impairment and 
Parkinsonism in patients with DLBD.   
1.2 Investigational Agent  
11C-PBR28 is a PET radioligand that binds to the 18 kDa translocator protein (TSPO), a 
marker of inflammation. 11C-PBR28 has previously been administered in humans.   
This radioligand will be administered in tracer doses. 11C-PBR28 will be administered at 
activity of up to 20 mCi per injection.   
1.3 Preclinical Data  
1.3.1. Preclinical Data for PBR28  
1.3.1.1. Pharmacology of PBR28  
1.3.1.1.1. General pharmacology  
Formerly called the peripheral benzodiazepine receptor, the 18 kDa translocator protein 
(TSPO) is located both in the central nervous system (CNS) and many peripheral organs, 
including endocrine tissues, kidney, heart, liver and blood cells. In the CNS, TSPO exists mostly 
but not exclusively in glial cells. In most of these organs, TSPO is located in the outer membrane 


 
  
 
 
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of mitochondria. In some organs, TSPO shows other subcellular localizations. For example, 
TSPO has been localized in mitochondrial inner membrane of guinea pig lung[30]. In rat liver, 
TSPO has been localized in two sites: a mitochondrial and an unidentified non-mitochondrial 
location[31]. In heart, TSPO is located in the plasma membrane, where it is reported to be 
coupled to calcium channels[32]. TSPO has been proposed to be involved in cellular 
proliferation, calcium channel activity, immune responses, transport of porphyrin and anion and 
regulation of steroid biosynthesis[33].   
PBR28 is a selective aryloxyanilide ligand for TSPO. PBR28 affinity was measured in 
using [3H]PK11195 as the radioligand and membranes prepared from rat brain, following a 
published procedure[34]. The Ki value was 0.2 nM in rat brain[35].  
  
1.3.1.1.2. In vitro receptor binding   
PBR28 showed a Ki of >10 μM at several central benzodiazepine (GABAA) receptors 
(α1β1γ2, α2β2γ2, α5β2γ2, and α6β2γ2 subtypes) and at 10 μM concentration was found to cause less 
than 50% displacement of reference ligand 5-HT1A,1B,1D,1E,2A−C,3,5A,6,7, α1A,1B,2A−C, β1−3, D1−4, H1−4, 
M2,5, DAT, NET, and SERT sites. A full displacement study revealed a Ki of 2.2 μM at κ opiate 
receptors[35].  
  
1.3.1.2. Animal toxicology for PBR28  
1.3.1.2.1. Rat study  
Male and female Sprague-Dawley rats (10/sex/group) were given a single intravenous 
(IV) dose of PBR28 at  440.5 µg/kg (2643 µg/m2, 500 times the human dose, Group 2) or at 88.1 
µg/kg (528.6 µg/m2, 100 times the human dose, Group 3) on Day 1 or a daily IV dose on Days 
1–5 of PBR28 at 88.1 µg/kg/day (528.6 µg/m2/day, 100 times the human dose, total dose 440.5 
µg/kg ). A control group (10/sex), Group 1, was given a single IV dose of vehicle, 5% ethanol in 
sterile saline, at an equivalent volume on Day 1. Dose administration in the repeat dose group 
(Group 4) was initiated 4 days prior to dosing of the single dose animals (Groups 1−3) so that 
necropsies occurred on the same calendar day for all groups. This schedule permitted control 
clinical pathology and necropsy data to be shared between the two dose regimens. Animals were 
sacrificed on Day 3 or Day 15 (interim and terminal necropsy, respectively) for the single dose 
groups (Groups 1–3) or on Day 7 or Day 19 (interim and terminal necropsy, respectively) for the 
repeat dose group (Group 4).   
The following parameters were evaluated: mortality/morbidity, clinical observations, 
body weights, food consumption, clinical pathology (hematology and serum chemistry), organ 
weights, macroscopic observation at necropsy and microscopic histopathology.   
All animals survived until their scheduled necropsy. No drug-related effects were found 
for clinical observations, body weights, food consumption, clinical pathology, organ weights, 
macroscopic or histopathologic evaluations.   
In conclusion, IV administration of PBR28 to male and female Sprague-Dawley rats at 
440.5 µg/kg (2643 µg/m2; 500× human dose) for a single day or 88.1 µg/kg (528.6 µg/m2, 100× 
human dose) for a single day or 5 days did not result in overt clinical adverse effects. The 
noobserved-adverse-effect (NOAEL) is considered to be at least 440.5 µg/kg (2643 µg/m2) for a 
single IV dose administration and at least 88.1 µg/kg/day (528.6 µg/m2/day) once each day for 5 
daily IV dose administrations. Because no significant adverse effects were seen at the highest 
dose evaluated, the maximum tolerated dose (MTD) could not be determined, but it is considered 


 
  
 
 
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to be greater than 440.5 µg/kg (2643 µg/m2) when given in a single IV injection and greater than 
88.1 µg/kg/day (528.6 µg/m2/day) when given in a daily IV injection for 5 consecutive days.   
  
1.3.1.2.2. Dog study  
This study involved three (Days 1, 8, and 15) dose administrations of PBR28 separated 
by one week intervals. Body weights were collected to determine dose calculations for each dose 
administration. Cardiovascular (CV) baseline data were collected prior to each dose 
administration including electrocardiogram (ECG), heart rate, respiratory rate, and blood 
pressure (indirect). Body temperature was also collected. Following administration of PBR28, 
each   
CV parameter was collected at 2, 30, 60, 120, 240 min, and 24 hr post dose administration.  
Clinical observations were made immediately post dose and daily thereafter for the duration of 
the study.    
The first IV administration of PBR28 was 26.43 µg/kg (528.6 µg/m2), 100 times the 
proposed human dose level. No adverse signs were observed for 7 days following administration. 
On Day 8, dose administration was escalated to 500 times the proposed human dose level 
(132.15 µg/kg; 2643 µg/m2). Again, no adverse clinical observations were observed for the week 
following administration. On Day 15, 1 naïve male and female dog were administered 132.15 
µg/kg PBR28 to confirm the NOAEL. Body weights and body temperatures were normal for all 
dogs for the duration of the study except for a minor body temperature decrease at the 24 hour 
time point for one dog administered 26.43 µg/kg PBR28 on Day 1. The minor change of body 
temperature for this dog is considered to be a sporadic variation and not test article-related.    
There were no test article related CV effects, including ST segment abnormalities, 
pronounced U waves, nor T wave changes such as changes in polarity, increased amplitude, or 
flattening seen in any of the dogs after a single intravenous injection of 100x or 500x the human 
dose level of PBR28 for the duration of the study, except for dog #4, which was administered 
132.15 µg/kg PBR28, had mild ST segment elevation on Day 15 at time points of 2 min and 4 
hour. Also in dog #4, there was a change in T wave polarity from pre-dose positive T wave to 2 
min negative T wave, to time 4 hour positive T wave, returning to negative T waves at time 24 
hour. Dog #3 had increased QRS amplitude at baseline, pre-dose and throughout the study; and 
this was not due to a treatment effect.   
   
The ST segment elevation changes seen in dog #4 administered 132.15 µg/kg PBR28 on 
Day 15 were transient, which may represent regional myocardial hypoxia and may have 
normalized at 24 hour post-dose. However, dog #3, also administered 132.15 µg/kg PBR28on 
Day 15, had no adverse test article related CV effects. In addition, Dogs #1 and #2 were 
administered 132.15 µg/kg PBR28 on Day 8, with no test article related CV effects; therefore, 
the changes seen in dog #4 were not definitively due to a treatment related adverse effect.   
Several dogs (#s 1, 2 and 4) had left and/or right mean electrical axis (MEA) deviations at 
the baseline, pre-dose and post dose administration. These changes can be affected by many 
extra-cardiac factors and are not considered to be test article-related.   
   
One dog (#2) had trivial hypotension at 4 hour post dose on Day 1. Animals with only 
reduced diastolic pressures were not considered to be hypotensive, as the diastolic pressure may 
be more likely to be erroneously low with oscillometric techniques; therefore, this effect is not 
considered to be test article related.    
In conclusion, the study results confirmed that there were no overt cardiovascular or 
toxicity effects of PBR28 after a single IV injection of 100 times the proposed human dose level 


 
  
 
 
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of PBR28 in Beagle dogs. Effects at 500 times the proposed human dose (132.15 µg/kg, 2643 
µg/m2) are inconclusive, with minor, transient effects on ST segment elevation seen in a single 
dog out of 4 evaluated at this dose level. The NOAEL for PBR28 is considered to be greater than  
26.43 µg/kg (528.6 µg/m2) and possibly less than 132.15 µg/kg (2643 µg/m2) via a single 
intravenous administration to male and female Beagle dogs. The maximum tolerated dose  
(MTD) was not determined, but is considered to be greater than 132.15 µg/kg (2643 µg/m2).  
  
1.3.1.3. Imaging studies with 11C-PBR28  
1.3.1.3.1. Whole body studies in monkey  
Male rhesus monkeys were scanned with 11C-PBR28. A preblocking scan was performed 
on 1 monkey after its baseline scan, in which nonradioactive PK 11195 was injected 3 min 
before the radioligand. Injection of 11C-PBR28 caused no change in ECG, heart, or respiration 
rates. Gallbladder, lungs, spleen, heart, brain, kidneys, liver, and urinary bladder were visually 
identified as organs with moderate-to-high activity. Uptake of 11C-PBR28 was highest in the 
lungs, with a peak of 50% injected activity occurring during the first frame (0–1.15 min). Peak 
uptake in kidneys, heart, liver, brain, and urinary bladder were 19.5%, 7%, 10%, 4.2%, and 0.8% 
standardized uptake value, respectively. The human effective dose estimated from monkey 
whole-body imaging was 10.3 μSv/MBq. The 3 organs with the highest radiation burden 
(μSv/MBq) were lungs (70.5), kidneys (43.1), and brain (19.5). Compared to baseline scans, the 
blocking experiment decreased areas under the curve in organs with PBRs (brain, lungs, kidneys, 
and heart) and increased areas under the curve in organs involved with metabolism and excretion 
(liver and bladder).  
  
1.3.1.3.1. Brain studies in monkey  
Rhesus monkeys had brain imaging with 11C-PBR28. A blocking experiment was 
performed by administering DAA1106 (3 mg/kg IV) with the radiotracer. In all brain scans, 
arterial blood sampling was performed to measure [11C]PBR28 and the radiometabolite levels in 
plasma.  
In the baseline experiments, [11C]PBR28 showed high peak uptake in brain (∼ 300% 
SUV) and was widely distributed, with greater activity in gray than white matter. The choroid 
plexus in the 4th ventricle had the highest radioactivity at late times, consistent with the known 
distribution of TSPO.  
In blocking experiments, brain activity at early times was increased due to elevated 
plasma concentrations of radiotracer. For example, the peak uptake of [11C]PBR28 was ∼ 300% 
SUV at baseline and ∼ 500% SUV after receptor blockade. Nevertheless, receptor blockade in 
brain was evident from the faster washout compared to baseline. That is, receptors were blocked 
by nonradioactive ligand, and the radioligand was not significantly retained in brain. For 
example, under baseline conditions, peak brain uptake occurred at about 40 min and decreased 
only 1.3% in the 15 min period thereafter. In contrast, for the receptor-blocked condition, brain 
uptake peaked at about 4 min and decreased by 62% in the subsequent 15 min.  
  
1.3.1.3.2. Rat stroke  
The ability of [11C]PBR28 PET to localize TSPO in a rat permanent middle cerebral 
artery occlusion (MCAO) model of neuroinflammation was determined. [11C]PBR28 was 
intravenously administered to rats at 4 and 7 days after permanent MCAO. In all experiments, 


 
  
 
 
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arterial blood was sampled for compartmental modeling of regional distribution volumes, and rat 
brains were sampled after imaging for in vitro [3H]PK 11195 autoradiography and histological 
evaluation. [11C]PBR28 PET and [3H]PK 11195 autoradiography showed similar areas of 
increased TSPO, especially in the peri-ischemic core. Results from these in vivo and in vitro 
methods were strongly correlated.  
  
1.4 Clinical Data to Date  
1.4.1. Human experience with PBR28  
  
Dr. Kreisl has 8 years experience using 11C-PBR28 in clinical studies at the National  
Institute of Mental Health Intramural Research Program. Over 100 subjects received injection of 
11C-PBR28 in clinical protocols on which he was either PI or sub-Investigator No AEs related to 
PBR28 administration have been reported.   
  
1.4.1.1. Whole body studies  
  
Whole-body images were acquired after intravenous bolus administration of 11C-PBR28 
in 7 healthy humans (651 +/- 111 MBq). For typical subjects, the 3 organs with highest exposure 
were those with the high TSPO densities (kidneys, spleen, and lungs), and the effective dose was 
6.6 microSv/MBq[36].  
  
1.4.1.2. Brain studies in controls  
  
Results from the first in-human brain study of 11C-PBR28 are as follows: Twelve healthy 
volunteers (1 female and 11 males, 25 ± 5 years of age, 81 ± 15 kg body weigh) underwent brain 
imaging with 11C-PBR28 with arterial catheterization[37]. Injected activity was 650 ± 92 MBq 
(17.6 ± 2.5 mCi). After injection of 11C-PBR28, 10 of 12 subjects showed moderate levels of 
activity in brain that washed out gradually. The peak uptake occurred at 5 min and was ∼ 200% 
SUV. Brain activity decreased to 50% of the peak by 70 min and to 40% of peak by 120 min. As 
expected from known distribution of TSPO in human brain, the distribution of activity was 
widespread and fairly uniform in gray matter of cerebral cortices and cerebellum, basal ganglia, 
and thalamus. The unconstrained two-tissue compartment model provided significantly better fit 
than the one-tissue compartment model, consistent with the presence of significant amounts of 
both specific and nonspecific binding in human brain.  
  
1.4.1.3. Differential affinity  
  
In the initial human whole-body and brain studies, two subjects were found to have a 
strikingly different time course of radioactivity in brain and periphery, including markedly faster 
washout[36, 37]. These two unusual subjects showed a higher peak radioactivity in brain at an 
earlier time. Activity washed quickly from brain and was at half of the peak concentration within 
only 6–7 min, compared to the other 10 subjects. In addition, brain activity remained almost 
constant after ∼ 20 min.  
  
In the whole body images, these unusual subjects had decreased activity in organs with 
high TSPO density—that is, brain, lungs, heart, spleen, and kidneys. In fact, spleen, lungs, and 
kidneys could not be identified visually. Furthermore, the activities in liver and gallbladder of 
these unusual subjects were much higher than in the other subjects.   


 
  
 
 
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One of the two subjects reported taking ibuprofen (1000 mg per day for several 
days/weeks) prior to the first scan. To test whether the ibuprofen may have blocked radioligand 
binding in brain, after ∼ 100 days off ibuprofen and 114 days after the first PET scan, we 
repeated the brain scan in this subject. The second scan was almost identical to the first in terms 
of peak uptake and washout rate.  
These unusual subjects were initially referred to as “non-binders,” reflecting the 
apparently lack of interaction between 11C-PBR28 and the target protein. In vitro binding assays 
using peripheral leukocytes isolated from blood samples taken from non-binders showed that 
these subjects expressed TSPO with ∼50-fold lower affinity for 11C-PBR28 than binders[38].  
 Later binding studies showed that there exist three patterns of 11C-PBR28 binding in human 
subjects[39, 40]. High affinity binders (HABs) behave as though they express only TSPO that 
binds with high affinity, low affinity binders (LABs) behave as though they express only TSPO 
that binds with low affinity, and mixed affinity binders (MABs) behave as though they express 
both a high- and low-affinity version of TSPO. Genetic studies determined there is a strong 
association between TSPO binding status and the rs6971 (Ala147Thr) single nucleotide 
polymorphism on the TSPO gene[29]. HABs lack this polymorphism, LABs are homozygous, 
and MABs are heterozygous. As the allelic frequency of the polymorphism is 30% based on 
population studies, 9% are expected to be LABs. Caucasians and African Americans have 
highest prevalence of the polymorphism while it is rare in Han Chinese and Japanese.    
Our ex 
vivo study showed that statistical correction for TSPO genotype improves the ability of 3H-
PBR28 to detect differences in TSPO density between controls and patients with schizophrenia in 
the dorsolateral prefrontal cortex[27]. Therefore, clinical studies have since employed the 
strategy of excluding LABs and performing statistical correction to remove the effect of 
systematic physiological differences in 11C-PBR28 binding between HABs and MABs.  
  
1.4.1.4. 11C-PBR28 in Alzheimer’s disease   
  
In our first AD PET study using 11C-PBR28, we found that AD patients had greater 
binding than MCI patients or controls[41]. The largest differences were seen in parietal and 
temporal regions. 11C-PBR28 binding correlated with clinical severity, brain atrophy, and earlier 
age of symptom onset. 11C-PBR28 binding did not conclusively correlate with amyloid burden 
as measured with 11C-Pittsburgh Compound B. This study used arterial blood sampling to 
calculate total distribution volume using the two-tissue compartment model, with correction for 
free fraction of radioligand. Even after TSPO genotype correction, variance of the PET data was 
relatively large (coefficient of variance = 34 - 73%).   
We later used a simplified ratio method to measure 11C-PBR28 binding in a larger sample 
(25 AD patients, 11 MCI patients, and 21 controls)[42]. By defining 11C-PBR28 binding as 
uptake in a target region divided by that in cerebellum (standardized uptake value ratio, SUVR), 
we were able to detect differences between AD patients and controls in more regions with larger 
effect sizes. Variance of the PET data was much lower. Cerebellum was chosen because it is a 
large brain region relatively spared in AD (particularly early in disease). This simplified ratio 
method removes the need for arterial sampling and reduces the amount of time needed on the 
PET scanner.  
  


 
  
 
 
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1.5 Dose Rationale and Risk/Benefits  
Based on prior human experience and human dosimetry data, we expect the proposed injected 
activity of 11C-PBR28 (up to 20 mCi) to be safe without expected toxicity.   
2 Study Objectives  
Primary objectives  
1). Determine feasibility of performing 11C-PBR28 PET in patients with DLBD.  
  
2). Determine effect size of 11C-PBR28 binding in patients with DLBD vs. controls.  
  
Our hypothesis is that 11C-PBR28 binding will be greater in DLBD patients than controls, 
with largest differences in temporal and limbic regions.   
  
Secondary objectives  
1). Identify potential reference regions for 11C-PBR28 binding in patients with DLBD.  
  
Our hypothesis is that correcting cortical 11C-PBR28 binding to that in the cerebellum, a region 
typically spared from pathology early in DLBD, will reduce intra-subject variance, avoid need for 
arterial catheterization, and improve sensitivity for detecting differences between DLBD patients 
and controls. We will also explore other potential reference regions such as cortical white matter.  
  
2). Determine if 11C-PBR28 binding correlates with cognitive impairment and Parkinsonism 
in patients with DLBD.  
  
We postulate that 11C-PBR28 binding will correlate with clinical severity (cognitive 
impairment and Parkinsonism) and the amount of atrophy on MRI.   
3 Study Design  
3.1 General Design  
  
Subject selection. Patients and controls will be recruited from the ADRC, CUMC clinics, 
and direct referrals. Target accrual is n = 4 completers in each group (DLBD and controls). ADRC 
subjects who have already undergone GBA and ApoE genotyping through ADRC participation 
will be preferentially enrolled. If possible, only previously-determined GBA mutation carriers will 
be included. However, if not enough GBA mutation carriers are recruited to meet target enrollment 
we will also enroll GBA non-carriers who are also known ApoE4 noncarriers. This strategy is 
designed to reduce the inclusion of patients with co-existent AD-related tangle pathology, as GBA 
mutation and ApoE4 non-carrier status are both associated with pure DLBD pathology[14-16]. If 
we are still unable to meet target enrollment then we will include patients of unknown GBA and 
ApoE genotype. However, these patients will have ApoE genotyping performed in this study, and 
we will perform post-hoc analysis to see if ApoE4 status affects 11C-PBR28 binding. ApoE testing 
will be done for research purposes only and results will not be disclosed to subjects. Subjects must 
have capacity to provide informed consent and consent will be obtained prior to study procedures.  
  


 
  
 
 
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Screening procedures. Subjects will undergo screening including history, physical 
examination, routine laboratory studies, neurological examination and brain MRI. Patients meeting 
clinical criteria for either dementia with Lewy bodies or Parkinson’s disease dementia (collectively 
considered as DLBD) will be included. Diagnosis will be made via consensus conference using 
clinical criteria[42]. Screening will also include genetic analysis to exclude subjects homozygous 
for the rs6971 (Ala147Thr) SNP on the TSPO gene. Homozygous subjects have negligible binding 
on 11C-PBR28 PET imaging and must be excluded; however, heterozygotes may be included[27, 
41]. Clinical evaluation will include Montreal Cognitive Assessment (MoCA), Clinical Dementia 
Rating scale – sum of boxes (CDR-SB), and Unified Parkinson’s Disease Rating Scale (UPDRS).   
  
PET imaging procedures. Subjects will undergo 11C-PBR28 PET in the CUMC Kreitchman 
PET center on a Siemens Biograph mCT. Prior to PET imaging, one arterial catheter and one 
intravenous catheter will be placed. After a low-dose CT scan for attenuation correction, up to 20 
mCi 11C-PBR28 will be injected intravenously over 60 seconds. Emission scan will begin at the 
start of the radioligand infusion according to the following frame schedule: 30 seconds x 6, 1 
minute x 3, 2 minutes x 2, and 5 minutes x 16 for total scan time of 90 min. During the emission 
scan, arterial samples will be drawn at 10 second intervals for the first 4 min using an automated 
blood sampler. Thereafter, manual arterial samples will be drawn at 6, 8, 10, 15, 20, 30, 40, 50, 60, 
75, and 90 minutes. Radioactivity in plasma will quantified by a γ-counter and analyzed by reverse-
phase chromatography to separate parent radioligand from radiometabolites[43] by the PET center 
radiometabolite laboratory. Free fraction of 11C-PBR28 in plasma (fP) will be measured by 
ultrafiltration and normalized using a standard derived from pooled donor plasma[44].   
  
Image analysis. PET images will be analyzed using two methods. First, kinetic analysis 
will be performed using PMOD software (PMOD Technologies). Subject MR images will be 
coregisered to native space PET images and then segmented to define gray matter regions of 
interest (ROIs): prefrontal cortex, combined superior and inferior parietal lobule, occipital cortex, 
inferior temporal cortex, combined middle and superior temporal cortex, hippocampus, 
parahippocampal gyrus, anterior cingulate, amygdala, insula, and cerebellum. These regions have 
been selected based on where the largest number of Lewy bodies have been previously 
reported[45]. The two-tissue kinetic model using time-activity curves from each ROI and the 
metabolite-corrected arterial input function will be used to calculate total distribution volume, 
corrected for free fraction of radioligand in plasma (VT/fP). Second, the cerebellum will be used as 
a pseudo-reference region to calculate standardized uptake value ratios (SUVR) for each target 
ROI. To determine the best time interval for non-invasive measurement of TSPO binding in DLBD, 
we will calculate SUVR using different time intervals and compute correlation coefficients with 
the gold-standard measurement VT/fP. The time intervals that correspond to SUVR values that have 
strongest correlation with VT/fP will be used to calculate SUVR for the group comparisons between 
patients with DLBD and controls.  
  
Statistical analysis.   
Primary outcome  
PET images will be analyzed using 1) the two-tissue kinetic model to calculate total distribution 
volume, corrected for free fraction of radioligand in plasma (VT/fP), and 2) the cerebellum as a 
pseudo-reference region to calculate standardized uptake value ratios (SUVR). Effect size will be 


 
  
 
 
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calculated for different regions as the difference in means (DLBD vs. controls) divided by the 
standard deviation.  
  
Secondary outcomes  
Coefficient of variation (%COV = standard deviation / mean) will be calculated for controls and 
patients with DLBD for both VT/fP and SUVR data. SUVR values will be calculated using 
cerebellum as reference regions. Additional reference regions will be explored. Correlation 
coefficients will be computed to look for correlation between 11C-PBR28 binding (VT/fP and 
SUVR) and clinical severity. Severity will be measured using CDR-sum of boxes, MoCA score, 
and UPDRS. We will also look for correlations between 11C-PBR28 binding and atrophy, as 
determined using ROI volumes derived from MRI data.  
  
3.2 Primary Study Endpoints  
  
Because the drug used in this study is a radioligand given at tracer doses, there are no clinical 
endpoints of the study.   
  
The primary outcome measures are 11C-PBR28 binding (using both VT/fP and SUVR). We expect 
binding will be greater in DLBD patients than in controls, with greatest difference in occipital, 
temporal, and parietal regions. We expect that 11C-PBR28 binding in cerebellum will not differ 
between DLBDpatients and controls and that this region can be used as a pseudo-reference 
region for non-invasive measurement of TSPO binding in DLBD. If we find that 11C-PBR28 
binding is different between patients and controls, we will explore other possible methods of 
non-invasive measurement. These methods include a cluster-based approach[46] or use of other 
potential pseudo-reference regions, such as white matter or whole gray matter.   
  
  
   
3.3 Secondary Study Endpoints  
N/A  
3.4 Primary Safety Endpoints  
N/A  
4 Subject Selection and Withdrawal  
4.1 Inclusion Criteria  
  
Inclusion criteria for patients:  
1. Age 60 and older  
2. Meet criteria for either a) dementia with Lewy bodies, or b) Parkinson’s disease dementia  
3. Able to provide informed consent   
4. Written and oral fluency in English   


 
  
 
 
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5. Able to participate in all scheduled evaluations and to complete all required tests and 
procedures.  
6. In the opinion of the investigator, the subject must be considered likely to comply with 
the study protocol and to have a high probability of completing the study.  
  
Inclusion criteria for controls:  
1. Age 60 and older  
2. Normal cognitive and motor function based on neurological examination  
3. Written and oral fluency in English   
4. Able to participate in all scheduled evaluations and to complete all required tests and 
procedures.  
5. In the opinion of the investigator, the subject must be considered likely to comply with 
the study protocol and to have a high probability of completing the study.  
  
Non-English speaking subjects will be excluded. The rationale is that because of the small scope 
of the study, we expect to reach accrual target with English speakers only. Also, because of the 
small scope we may not have resources for providing Spanish-speaking staff at all study visits.  
4.2 Exclusion Criteria  
1. Past or present history of certain brain disorders (other than DLB or PDD in the case of 
patient participants).  
2. Certain significant medical conditions, which make study procedures of the current study 
unsafe. Such serious medical conditions include uncontrolled epilepsy and multiple 
serious injuries.   
3. Contraindication to MRI scanning  
4. Conditions precluding entry into the scanners (e.g. morbid obesity, claustrophobia, etc.).  
5. Exposure to research related radiation in the past year that, when combined with this 
study, would place subjects above the allowable limits.  
6. Low affinity binding on TSPO genetic screen  
7. Currently taking anticoagulant drugs (e.g., warfarin)  
8. Women who are able to become pregnant   
  
4.3 Subject Recruitment and Screening  
  
Patients and controls will be recruited from the ADRC, CUMC clinics, and direct 
referrals. Approximately 50% of enrolled subjects are expected to fail screening procedures or 
withdraw from the study. We therefore plan to screen 8 subjects in each group to ensure at least 4 
in each group complete study procedures. Screening will also include genetic analysis to exclude 
subjects homozygous for the rs6971 (Ala147Thr) single nucleotide polymorphism on the TSPO 
gene.   
  


 
  
 
 
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4.4 Early Withdrawal of Subjects  
4.4.1 When and How to Withdraw Subjects  
Subjects will be withdrawn if they develop serious medical illness during the study, defined as an 
event determined to be grade 3 or higher, i.e., severe or life-threatening. The exception is a single 
event of syncope if felt by the PI to be vasovagal in etiology and related to catheter placement or 
venipuncture. Vasovagal syncope in response to blood drawing and IV placement is common 
and does not warrant study withdrawal if the subject’s loss of consciousness is brief (i.e., less 
than 5 minutes) and does not require further medical attention.  
4.4.2 Data Collection and Follow-up for Withdrawn Subjects  
Data collected prior to withdrawal will be analyzed if possible. No data collection will take place 
after withdrawal in subjects who drop out of the study.   
5 Study Drug   
5.1 Description  
11C-PBR28 is a PET radioligand that binds to the 18 kDa translocator protein. 11C-PBR28 will be 
administered at tracer doses and is not expected to have a pharmacological effect.  
  
5.2 Treatment Regimen  
N/A  
5.3 Method for Assigning Subjects to Treatment Groups  
N/A  
5.4 Preparation and Administration of Study Drug  
11C-PBR28 will be synthesized and administered by the CUMC PET Department. Injection will 
be administered at a target imaging dose of up to 20 mCi (740 MBq).  
  
Calculation of activity to be injected will be performed by a qualified radiopharmacist in the 
CUMC PET department.  
5.5 Subject Compliance Monitoring  
N/A  
5.6 Prior and Concomitant Therapy  
Subjects will not be included if they are currently taking anticoagulants such as warfarin. 
Antiplatelet drugs such as aspirin are allowed.  
  
5.7 Packaging  
N/A  


 
  
 
 
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5.8 Blinding of Study Drug  
N/A   
5.9 Receiving, Storage, Dispensing and Return  
5.9.1 Receipt of Drug Supplies  
Upon receipt of the of the study treatment supplies, the CUMC PET Center staff will perform 
an inventory and a drug receipt log will be filled out and signed by the person accepting the 
shipment.  Any damaged or unusable study drug in a given shipment (active drug or 
comparator) will be documented in the study files.   
5.9.2 Storage  
N/A  
5.9.3 Dispensing of Study Drug  
11C-PBR28 will be administered the same day of synthesis.  
5.9.4 Return or Destruction of Study Drug  
Any unused radiopharmaceutical will be on site and documented in the study files.  
6 Study Procedures  
  
This study will involve up to 3 outpatient visits. See schedule below.  
  
At the first screening visit, subjects will sign informed consent, undergo history and physical and 
neurological examination, and have blood drawn for routine safety laboratories. Menopause 
(defined as absence of menses for at least one year) or history of hysterectomy and/or tubal 
ligation will be confirmed by medical history to exclude women of child-bearing potential. 
Blood will also be drawn for TSPO, ApoE and GBA genotyping if not previously performed in 
another study.   
  
Brain MRI will take place at the Neurological Institute Neuro MRI Center on 3T Philips scanner. 
MRI will be performed to rule out intracranial disease other than PDD or DLB, and for 
coregistration with the PET images for analysis. An additional screening visit is required for 
brain MRI; however, this may be performed on the same day as the first screening visit is 
scheduling allows. Up to 2 screening visits are therefore anticipated. However, additional visits 
may be necessary if certain tests must be scheduled on different days due to scheduling. If 
subjects require a sedative for the MRI (e.g., because of claustrophobia), they may receive a 
onetime dose of lorazepam (0.5 - 1 mg orally) or equivalent dose of an alternative 
benzodiazepine prior to the MRI. Side effects of anti-anxiety medications may include prolonged 
sedation (usually less than 8 hours), lightheadedness, motor incoordination, difficulties with 
balance, or confusion. These effects are temporary and go away after several hours. Subjects will 
be instructed not to drive, handle machinery, or drink alcohol until the next day. Subjects who 
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visit, and will be accompanied by a member of the study staff during the MRI. The PI will be 
available in the event of an adverse event during the MRI.  
  
One study visit will be required for the 11C-PBR28 PET scan. At the PET scan visit, one IV 
catheter will be placed by a qualified clinician for purposes of radioligand injection. After 
performing an Allen test to ensure ulnar perfusion to the hand, a catheter will be placed in the 
radial artery by a qualified clinician. After a CT attenuation scan, up to 20 mCi 11C-PBR28 will 
be injected via the IV. The emission scan will last 90 minutes, during which members of the 
study team will draw arterial blood samples.   
  
11C-PBR28 PET scans must be completed within 12 months of the brain MRI. Other screening 
procedures must be done within 60 days prior to the PET scan.  
Study Procedures  
Screen a  
Study b  
Day -60 to 1  
Visit 1  
Informed consent  
X  
   
Medical history   
X  
   
Inclusion/exclusion criteria  
X  
X  
Blood draw for genetic tests  
X  
   
MRI  
X  
   
11C-PBR28 PET  
   
X  
Laboratory determinations   
X  
   
Neurological examination  
X  
   
Vital signs   
X  
   
Height and weight  
X  
 
Adverse events   
X  
X  
 
 


 
  
 
 
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7 Statistical Plan  
7.1 Sample Size Determination  
This study is intended to generate pilot data as 11C-PBR28 PET imaging has not previously 
been performed in patients with diffuse Lewy body disease. Therefore, preliminary data to 
accurately power this study does not exist. However, our previous study using 11C-PBR28 in 
patients with Alzheimer’s disease (a different neurodegenerative disease) showed effect size of 
1.5 in brain regions with largest difference between patients and controls. If a similar effect size 
is seen in patients with diffuse Lewy body disease then we would expect statistically significant 
differences with sample size of n = 8.  
  
7.2 Statistical Methods  
Primary outcome  
PET images will be analyzed using 1) the two-tissue kinetic model to calculate total distribution 
volume, corrected for free fraction of radioligand in plasma (VT/fP), and 2) the cerebellum as a 
pseudo-reference region to calculate standardized uptake value ratios (SUVR). Effect size will be 
calculated for different regions as the difference in means (DLBD vs. controls) divided by the 
standard deviation.  
  
Secondary outcomes  
Coefficient of variation (%COV = standard deviation / mean) will be calculated for controls and 
patients with DLBD for both VT/fP and SUVR data. SUVR values will be calculated using 
cerebellum as reference regions. Additional reference regions will be explored. Correlation 
coefficients will be computed to look for correlation between 11C-PBR28 binding (VT/fP and 
SUVR) and clinical severity. Severity will be measured using CDR-sum of boxes, MoCA score, 
and UPDRS. We will also look for correlations between 11C-PBR28 binding and atrophy, as 
determined using ROI volumes derived from MRI data.  
7.3 Subject Population(s) for Analysis  
Subjects will be age 60 or older. Healthy controls and subjects who meet clinical criteria for DLB 
or PDD will be included.  
8 Safety and Adverse Events  
8.1 Definitions  
Adverse Event  
An adverse event (AE) is any symptom, sign, illness or experience that develops or worsens 
in severity during the course of the study.  Intercurrent illnesses or injuries should be regarded 
as adverse events.  Abnormal results of diagnostic procedures are considered to be adverse 
events if the abnormality:  
• results in study withdrawal  


 
  
 
 
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• is associated with a serious adverse event  
• is associated with clinical signs or symptoms  
• leads to additional treatment or to further diagnostic tests  
• is considered by the investigator to be of clinical significance Serious Adverse 
Event  
Adverse events are classified as serious or non-serious.  A serious adverse event is any AE 
that is:   
• fatal  
• life-threatening  
• requires or prolongs hospital stay  
• results in persistent or significant disability or incapacity  
• an important medical event  
  
Important medical events are those that may not be immediately life threatening, but are 
clearly of major clinical significance.   They may jeopardize the subject, and may require 
intervention to prevent one of the other serious outcomes noted above.  For example, drug 
overdose or abuse, a seizure that did not result in in-patient hospitalization, or intensive 
treatment of bronchospasm in an emergency department would typically be considered 
serious.   
  
All adverse events that do not meet any of the criteria for serious should be regarded as 
non-serious adverse events.   
Adverse Event Reporting Period  
The study period during which adverse events must be reported is normally defined as the 
period from the initiation of any study procedures to the end of the study treatment 
followup.  For this study, the study drug follow-up is defined as 24 hours following the last 
administration of study drug.   
Preexisting Condition  
A preexisting condition is one that is present at the start of the study.  A preexisting 
condition should be recorded as an adverse event if the frequency, intensity, or the 
character of the condition worsens during the study period.  
General Physical Examination Findings  
At screening, any clinically significant abnormality should be recorded as a preexisting 
condition.  At the end of the study, any new clinically significant findings/abnormalities 
that meet the definition of an adverse event must also be recorded and documented as an 
adverse event.   
Post-study Adverse Event  
All unresolved adverse events should be followed by the investigator until the events are 
resolved, the subject is lost to follow-up, or the adverse event is otherwise explained.  At 
the last scheduled visit, the investigator should instruct each subject to report any 
subsequent event(s) that the subject, or the subject’s personal physician, believes might 
reasonably be related to participation in this study.  The investigator should notify the study 
sponsor of any death or adverse event occurring at any time after a subject has discontinued 


 
  
 
 
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or terminated study participation that may reasonably be related to this study.  The sponsor 
should also be notified if the investigator should become aware of the development of 
cancer or of a congenital anomaly in a subsequently conceived offspring of a subject that 
has participated in this study.   
Abnormal Laboratory Values  
A clinical laboratory abnormality should be documented as an adverse event if any one of 
the following conditions is met:   
• 
The laboratory abnormality is not otherwise refuted by a repeat test to confirm the 
abnormality  
• 
The abnormality suggests a disease and/or organ toxicity  
• 
The abnormality is of a degree that requires active management; e.g. change of 
dose, discontinuation of the drug, more frequent follow-up assessments, further 
diagnostic investigation, etc.  
Hospitalization, Prolonged Hospitalization or Surgery  
Any adverse event that results in hospitalization or prolonged hospitalization should be 
documented and reported as a serious adverse event unless specifically instructed otherwise 
in this protocol.  Any condition responsible for surgery should be documented as an 
adverse event if the condition meets the criteria for and adverse event.   
  
Neither the condition, hospitalization, prolonged hospitalization, nor surgery are reported 
as an adverse event in the following circumstances:  
• 
Hospitalization or prolonged hospitalization for diagnostic or elective surgical 
procedures for a preexisting condition.  Surgery should not be reported as an 
outcome of an adverse event if the purpose of the surgery was elective or diagnostic 
and the outcome was uneventful.  
• 
Hospitalization or prolonged hospitalization required to allow efficacy 
measurement for the study.  
• 
Hospitalization or prolonged hospitalization for therapy of the target disease of the 
study, unless it is a worsening or increase in frequency of hospital admissions as 
judged by the clinical investigator.  
8.2 Recording of Adverse Events  
At each contact with the subject, the investigator must seek information on adverse events (AEs) 
by specific questioning and, as appropriate, by examination.  Information on all adverse events 
should be recorded immediately in the source document, and also in the appropriate adverse 
event module of the case report form (CRF).  All clearly related signs, symptoms, and abnormal 
diagnostic procedures results should recorded in the source document, though should be grouped 
under one diagnosis.  
  
All adverse events occurring during the study period must be recorded.  The clinical course of 
each event should be followed until resolution, stabilization, or until it has been determined that 
the study treatment or participation is not the cause.  Serious adverse events that are still ongoing 


 
  
 
 
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at the end of the study period must be followed up to determine the final outcome.  Any serious 
adverse event that occurs after the study period and is considered to be possibly related to the 
study treatment or study participation should be recorded and reported immediately.  
  
As per CUMC IRB’s Policy, Unanticipated Problems will be defined as follows:  
  
Unanticipated Problem (UP) is any incident, experience or outcome involving risk to subjects or 
others in any human subjects research that meets all of the following criteria:  
  
• 
Unexpected (in terms of nature, severity or frequency) given (a) the research procedures 
that are described in the IRB-approval protocol and informed consent document and (b) 
the characteristics of the subject population being studied;  
• 
Related or possibly related to participation in such research (i.e., there is a reasonable 
possibility that the incident, experience or outcome may have been caused by the 
procedures involved in such research); and  
• 
Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic or social harm) than was previously known or 
recognized (HHS IRB Guidance, Section I).  
  
The following AEs will be considered Unexpected Problems:  
  
• 
A single occurrence of a serious, unexpected event that is uncommon and strongly 
associated with drug exposure  
• 
A single occurrence, or a small number of occurrences, of a serious, unexpected event 
that is not commonly associated with drug exposure  
• 
Multiple occurrences of an AE that, based on an aggregate analysis, are determined not to 
be isolated occurrences and involve risk to human subjects  
• 
An AE that is described or addressed in the investigator's brochure, protocol or informed 
consent documents (a Described AE), but occurs at a specificity or severity that is 
inconsistent with prior observations  
• 
A serious Described AE, but for which the rate of occurrence represents a clinically 
significant increase in the expected rate of occurrence  
• 
Any other AE or safety finding that would cause the sponsor to modify the investigator's 
brochure, study protocol or informed consent documents or would prompt other action by 
the IRB to ensure the protection of human subjects (2009 FDA Guidance, Section III 
(A)).  
8.3 Reporting of Serious Adverse Events  
8.3.1 IRB Notification by Investigator  
At the time of the Occurrence of an Unanticipated Problem (UP):  
  
Each UP will be reported to the IRB, whether or not (a) it is serious or non-serious or (b) 
it occurs at a site at which the PI is conducting the research.  


 
  
 
 
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The UP will be reported promptly, but not later than one week following the occurrence 
of the UP or the PI's acquiring knowledge of the UP.  
  
The PI will make the determination as to whether an incident, experience or outcome 
constitutes a UP.  
  
Each Unanticipated Problem will be reported to the IRB using the Unanticipated Problem 
Report module in Rascal.  
  
The investigator must conclude in the Unanticipated Problem Report whether the protocol 
and/or consent form(s) should be modified as the result of the UP.  If the protocol and/or 
consent document(s) requires a revision, a modification must be submitted in Rascal.  
  
At the Time of Continuing Review of a Protocol:  
  
At the time of continuing review of a protocol, the PI will submit a summary of all UPs 
that occurred during the review period and since the beginning of the study.  The summary 
for each UP should include:  
  
• 
The number of subjects who experienced the UP;  
• 
The investigator's determination as to whether or not the UP was serious;  
• 
The investigator's determination as to the UP's relationship to the study 
procedures (e.g., definitely related, probably related or possibly related).  
8.3.2 FDA Notification by Investigator  
The principal investigator shall notify the FDA by telephone or by facsimile transmission 
of any unexpected fatal or life-threatening experience associated with the use of the drug 
as soon as possible but no later than 7 calendar days from the investigator’s original receipt 
of the information.  
  
If a previous adverse event that was not initially deemed reportable is later found to fit the 
criteria for reporting, the principal investigator will submit the adverse event in a written 
report to the FDA as soon as possible, but no later than 15 calendar days from the time the 
determination is made.  
8.4 Unblinding Procedures  
N/A  
8.5 Stopping Rules   
N/A  


 
  
 
 
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8.6 Medical Monitoring  
We will not have a specific Data and Safety Monitoring Board. The PI will be monitoring the 
study. Adverse events will be documented in the patient’s chart. Notification of UPs to the IRB 
and FDA will take place as described in Section 8.3.   
  
9 Data Handling and Record Keeping  
9.1 Confidentiality  
Information about study subjects will be kept confidential and managed according to the 
requirements of the Health Insurance Portability and Accountability Act (HIPAA).  Those 
regulations require a signed subject authorization informing the subject of the following:   
• 
What protected health information (PHI) will be collected from subjects in this study  
• 
Who will have access to that information and why  
• 
Who will use or disclose that information  
• 
The rights of a research subject to revoke their authorization for use of their PHI.   
In the event that a subject revokes authorization to collect or use PHI, the investigator, by 
regulation, retains the ability to use all information collected prior to the revocation of subject 
authorization.  For subjects that have revoked authorization to collect or use PHI, attempts 
should be made to obtain permission to collect at least vital status (i.e. that the subject is alive) at 
the end of their scheduled study period.  
9.2 Source Documents  
Source data is all information, original records of clinical findings, observations, or other activities 
in a clinical trial necessary for the reconstruction and evaluation of the trial.  Source data are 
contained in source documents   Examples of these original documents, and data records include: 
hospital records, clinical and office charts, laboratory notes, memoranda, subjects’ diaries or 
evaluation checklists, pharmacy dispensing records, recorded data from automated instruments, 
copies or transcriptions certified after verification as being accurate and complete, microfiches, 
photographic negatives, microfilm or magnetic media, x-rays, subject files, and records kept at the 
pharmacy, at the laboratories, and at medico-technical departments involved in the clinical trial.  
9.3 Case Report Forms  
The study case report form (CRF) is the primary data collection instrument for the study.  All data 
requested on the CRF must be recorded.  All missing data must be explained.  If a space on the 
CRF is left blank because the procedure was not done or the question was not asked, write “N/D”.  
If the item is not applicable to the individual case, write “N/A”.  All entries should be printed 
legibly in black ink.  If any entry error has been made, to correct such an error, draw a single 
straight line through the incorrect entry and enter the correct data above it.  All such changes must 
be initialed and dated.  DO NOT ERASE OR WHITE OUT ERRORS.  For clarification of illegible 
or uncertain entries, print the clarification above the item, then initial and date it.  
  


 
  
 
 
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10 Study Monitoring, Auditing, and Inspecting  
10.1 Study Monitoring Plan  
This study will be monitored by the Principal Investigator. Dr. Kreisl will submit annual progress 
reports to the FDA within 60 days of the anniversary of the date that the IND became active (the 
date clinical studies were permitted to begin) in accordance with 21 CFR 312.33.  
11 Ethical Considerations  
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 guidelines), 
applicable government regulations and Institutional research policies and procedures.  
  
This protocol and any amendments will be submitted to a properly constituted independent 
Institutional Review Board (IRB), in agreement with local legal prescriptions, for formal 
approval of the study conduct.  The decision of the IRB concerning the conduct of the study will 
be made in writing to the investigator and a copy of this decision will be provided to the sponsor 
before commencement of this study.  The investigator should provide a list of EC/IRB members 
and their affiliate to the sponsor.  
  
All subjects for this study will be provided a consent form describing this study and providing 
sufficient information for subjects to make an informed decision about their participation in this 
study.  See attached copy of the Subject Informed Consent Form.  This consent form will be 
submitted with the protocol for review and approval by the IRB for the study.  The formal 
consent of a subject, using the IRB-approved consent form, must be obtained before that subject 
is submitted to any study procedure.  This consent form must be signed by the subject and the 
investigator-designated research professional obtaining the consent.   
  
12 Study Finances  
12.1 Funding Source  
This study is funded by a Rose and Boris Katz Assistant Professorship endowed to Dr. Kreisl and 
a Pilot Grant Award from the Alzheimer's Disease Research Center (ADRC) of the Taub 
Institute for Research on Alzheimer's Disease and the Aging Brain (PI = Kreisl).   
12.2 Conflict of Interest  
Any investigator who has a conflict of interest with this study (patent ownership, royalties, or 
financial gain greater than the minimum allowable by their institution, etc.) must have the 
conflict reviewed by a properly constituted Conflict of Interest Committee with a 
Committeesanctioned conflict management plan that has been reviewed and approved by the 
study sponsor prior to participation in this study.  All Columbia University Medical Center 
investigators will follow the University conflict of interest policy.  


 
  
 
 
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12.3 Subject Stipends or Payments  
Subjects will receive compensation commensurate to other studies at CUMC. Subjects will 
receive payment of $50 for the screening procedures and $200 for the PET scan ($250 total).  
13 Publication Plan  
Neither the complete nor any part of the results of the study carried out under this protocol, nor 
any of the information provided by the sponsor for the purposes of performing the study, will be 
published or passed on to any third party without the consent of the study sponsor.  Any 
investigator involved with this study is obligated to provide the sponsor with complete test 
results and all data derived from the study.  
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