Targeting Cellular Senescence With Senolytics to Improve Skeletal Health in Older Humans: A Phase 2, Single-Center, 20-week, Open-Label, Randomized Controlled Trial.
Phase: PHASE2
Conditions: Healthy
Sponsor: Sundeep Khosla, M.D.

 
 
Targeting Cellular Senescence with Senolytics 
to Improve Skeletal Health in Older Humans: A 
Phase 2, Single-Center, 20-week, Open-Label, 
Randomized Controlled Trial 
 
NCT04313634 
 
12/8/2023 

Effects of Senolytics on Markers of Aging and Bone Metabolism  
 
 
 
 
 
 
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Targeting Cellular Senescence with Senolytics to Improve Skeletal Health in Older 
Humans: A Phase 2, Single-Center, 20-Week, Open-Label, Randomized Controlled Trial 
 
 
 
 
 
Principal Investigators and 
Regulatory Sponsor: 
Sundeep Khosla, MD 
Division of Endocrinology and Kogod Center on Aging 
Mayo Clinic 
200 First Street Southwest 
Rochester, MN  55905 
 
 
Co-Principal Investigator: 
Joshua Farr, PhD  
Division of Endocrinology and Kogod Center on Aging 
Mayo Clinic 
200 First Street Southwest 
Rochester, MN  55905 
 
 
Study Product: 
• Sprycel® (dasatinib) and Quercetin 
 
 
Protocol Number: (IRBe) 
18-010546 
 
IND Number: 
145558 
 
 
 
 
 
 
September 10, 2019; Version 2.0 
December 9, 2019; Version 3.0 
February 19, 2020; Version 4.0 
May 19, 2020; Version 5.0 
July 14, 2020; Version 6.0 
November 20. 2020; Version 7.0 
March 4, 2021; Version 8.0 
May 20, 2021; Version 9.0 
July 16, 2021; Version 10.0 
January 7, 2022; Version 11.0 
May 31, 2022; Version 12.0 
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March 22, 2023; Version 14.0 
 

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Table of Contents 
 
 
 
STUDY SUMMARY ...................................................................................................................................... 5 
1 
INTRODUCTION ................................................................................................................................. 6 
1.1 
BACKGROUND .................................................................................................................................... 6 
1.2 
INVESTIGATIONAL AGENT .................................................................................................................... 7 
1.3 
PRECLINICAL DATA ............................................................................................................................. 8 
CLINICAL DATA TO DATE ........................................................................................................................... 11 
1.4 
DOSE RATIONALE.............................................................................................................................. 12 
1.5 
RISKS AND BENEFITS ......................................................................................................................... 13 
2 
STUDY OBJECTIVES ........................................................................................................................ 14 
3 
STUDY DESIGN ................................................................................................................................. 15 
3.1 
GENERAL DESCRIPTION ..................................................................................................................... 15 
3.2 
NUMBER OF SUBJECTS ....................................................................................................................... 16 
3.3 
DURATION OF PARTICIPATION ............................................................................................................ 16 
3.4 
PRIMARY STUDY ENDPOINTS .............................................................................................................. 16 
3.5 
SECONDARY STUDY ENDPOINTS ......................................................................................................... 17 
3.6 
PRIMARY SAFETY ENDPOINTS ............................................................................................................ 17 
3.7 
IDENTIFICATION OF SOURCE DATA ...................................................................................................... 17 
4 
SUBJECT SELECTION ENROLLMENT AND WITHDRAWAL ...................................................... 17 
4.1 
INCLUSION CRITERIA ......................................................................................................................... 17 
4.2 
EXCLUSION CRITERIA ........................................................................................................................ 17 
4.3 
SUBJECT RECRUITMENT, ENROLLMENT AND SCREENING ....................................................................... 20 
4.4 
EARLY WITHDRAWAL OF SUBJECTS .................................................................................................... 21 
4.4.1 
When and How to Withdraw Subjects ........................................................................................ 21 
4.4.2 
Data Collection and Follow-up for Withdrawn Subjects .............................................................. 22 
5 
STUDY DRUG .................................................................................................................................... 22 
5.1 
DESCRIPTION .................................................................................................................................... 22 
5.2 
TREATMENT REGIMEN ....................................................................................................................... 23 
5.3 
METHOD FOR ASSIGNING SUBJECTS TO TREATMENT GROUPS ................................................................ 23 
5.4 
PREPARATION AND ADMINISTRATION OF STUDY DRUG ......................................................................... 24 
5.5 
SUBJECT COMPLIANCE MONITORING ................................................................................................... 24 
5.6 
PRIOR AND CONCOMITANT THERAPY .................................................................................................. 24 
5.7 
PACKAGING ...................................................................................................................................... 24 
5.8 
MASKING/BLINDING OF STUDY........................................................................................................... 24 
5.9 
RECEIVING, STORAGE, DISPENSING AND RETURN ................................................................................. 24 
5.9.1 
Receipt of Drug Supplies .......................................................................................................... 24 
5.9.2 
Storage ................................................................................................................................... 24 
5.9.3 
Dispensing of Study Drug ......................................................................................................... 25 
5.9.4 
Return or Destruction of Study Drug ......................................................................................... 25 
6 
STUDY PROCEDURES ...................................................................................................................... 26 
6.1 
VISIT 1 – SCREENING VISIT ................................................................................................................ 26 
6.2 
VISIT 2 – BASELINE/ENROLLMENT (START OF WEEK 1, DAY 1) .............................................................. 26 
6.3 
VISIT 3 – START OF WEEK #3 (DAY 15 ± 5 DAYS) ................................................................................. 27 
6.4 
VISIT 4 – START OF WEEK #5 (DAY 29 ± 5 DAYS) ................................................................................. 28 
6.5 
VISIT 5 – START OF WEEK #9 (DAY 57 ± 5 DAYS) ................................................................................. 28 

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6.6 
VISIT 6 – START OF WEEK #13 (DAY 85 ± 5 DAYS) ............................................................................... 29 
6.7 
VISIT 7 – START OF WEEK #17 (DAY 113 ± 5 DAYS) ............................................................................. 29 
6.8 
VISIT 8-END OF WEEK #20 (DAY 140 ± 5 DAYS) ........................................................................... 31 
6.9 ASSESSMENTS ..................................................................................................................................... 29 
6.9.1 
Examinations and Procedure .................................................................................................... 30 
6.9.2 
Laboratory Assessments ........................................................................................................... 30 
6.9.3 
Questionnaires: ....................................................................................................................... 31 
6.9.4 
Skin AGEs Assessment: ............................................................................................................ 31 
7 
STATISTICAL PLAN ......................................................................................................................... 31 
7.1 
SAMPLE SIZE DETERMINATION ........................................................................................................... 31 
7.2 
STATISTICAL METHODS ..................................................................................................................... 32 
7.3 
SUBJECT POPULATION(S) FOR ANALYSIS ............................................................................................. 33 
8 
SAFETY AND ADVERSE EVENTS ................................................................................................... 33 
8.1 
DEFINITIONS ..................................................................................................................................... 33 
8.2 
RECORDING OF ADVERSE EVENTS ....................................................................................................... 35 
8.3 
REPORTING OF SERIOUS ADVERSE EVENTS AND UNANTICIPATED PROBLEMS .......................................... 36 
8.3.1 
Sponsor-Investigator reporting: notifying the Mayo IRB ............................................................. 36 
8.3.2 
Sponsor-Investigator reporting: Notifying the FDA ..................................................................... 36 
8.4 
UNMASKING/UNBLINDING PROCEDURES ............................................. ERROR! BOOKMARK NOT DEFINED. 
8.5 
STOPPING RULES ............................................................................................................................... 37 
8.6 
MEDICAL MONITORING ..................................................................................................................... 37 
8.6.1 
Internal Data and Safety Monitoring Board ............................................................................... 37 
8.6.2 
Independent Data and Safety Monitoring Board ......................................................................... 38 
9 
DATA HANDLING AND RECORD KEEPING .................................................................................. 38 
9.1 
CONFIDENTIALITY ............................................................................................................................. 38 
9.2 
SOURCE DOCUMENTS ........................................................................................................................ 38 
9.3 
CASE REPORT FORMS ........................................................................................................................ 38 
9.4 
RECORDS RETENTION ........................................................................................................................ 39 
10 
STUDY MONITORING, AUDITING, AND INSPECTING ................................................................ 39 
10.1 
STUDY MONITORING PLAN ............................................................................................................ 39 
10.2 
AUDITING AND INSPECTING ........................................................................................................... 40 
11 
ETHICAL CONSIDERATIONS ......................................................................................................... 40 
12 
STUDY FINANCES ............................................................................................................................ 40 
12.1 
FUNDING SOURCE ......................................................................................................................... 40 
12.2 
CONFLICT OF INTEREST ................................................................................................................. 40 
12.3 
SUBJECT STIPENDS OR PAYMENTS .................................................................................................. 40 
13 
PUBLICATION PLAN ....................................................................................................................... 41 
14 
REFERENCES ................................................................................................................................... 41 
 
 
 
 
 
 
 
 

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List of Abbreviations 
 
 
LIST OF ABBREVIATIONS 
AE 
Adverse Event/Adverse Experience 
CFR 
Code of Federal Regulations 
CRF 
Case Report Form 
DSMB 
Data and Safety Monitoring Board 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
HIPAA 
Health Insurance Portability and Accountability Act 
IB 
Investigator’s Brochure 
IND 
Investigational New Drug Application 
IRB 
Institutional Review Board 
PHI 
Protected Health Information 
PI 
Principal Investigator 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SOP 
Standard Operating Procedure 
 
 
 
 
 
 
 

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Study Summary 
Title 
Targeting Cellular Senescence with Senolytics to Improve Skeletal 
Health in Older Humans: A Phase 2, Single-Center, 20-Week, Open- 
Label, Randomized Controlled Trial 
Running Title 
Effects of Senolytics on Markers of Aging and Bone Metabolism 
Protocol Number 
18-010546 
Phase 
Phase II 
Methodology 
Randomized, open-label trial 
Overall Study 
Duration 
20 weeks 
Subject Participation 
Duration 
20 weeks, plus screening visit 
Single or Multi-Site  
Single Site Study 
Objectives 
Test whether intermittent senolytic therapy reduces markers of 
biological age and improves bone turnover markers and skeletal 
parameters in older postmenopausal women 
Number of Subjects 
240 screened, 74 accrued 
Diagnosis and Main 
Inclusion Criteria 
Elderly postmenopausal women, aged ≥60years, with increased 
senescent cell abundance in blood 
Study Product, Dose, 
Route, Regimen 
This study will involve an oral treatment regimen with Dasatinib (D; 
100 mg for two days) plus Quercetin (Q; 1000 mg total daily for three 
consecutive days) taken orally on an intermittent schedule (starting 
every 28 days) with no-therapy periods in between dosing regimens, 
repeated every 28 days over 20 weeks, resulting in five total dosing 
periods throughout the entire intervention. Note that we are modifying 
the protocol to complete 30 subjects in the control group, 30 in the 
D+Q group, and 14 who have completed the F-group but no additional 
subjects will be added to that group. 
Duration of 
Administration 
Five total intermittent dosing periods throughout the 20-week 
intervention 
Reference therapy 
Untreated control group 
Statistical 
Methodology 
Using previously published data, power calculations were conducted, 
while allowing for an estimated withdrawal rate of 10%. This 
calculation demonstrated that 30 subjects per group will provide the 
study with 90% power to detect a 19.8% difference in the bone 
resorption marker, CTX (17.1% with 80% power), which is our 
primary endpoint.  
 

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1 Introduction 
 
This document is a protocol for a human research study. This study will be carried out in 
accordance with the applicable United States government regulations and Mayo Clinic research 
policies and procedures. 
1.1 
Background 
 
The Mayo Clinic Aging Group is at the forefront of discovering, developing, and optimizing 
therapies to eliminate senescent cells – “senolytics”. With aging, DNA damage and/or other 
cellular stressors(1-4) cause proliferating(5,6) and terminally differentiated, non-dividing cells(7-10) 
to undergo senescence. Senescent cell characteristics include profound chromatin and secretome 
changes, increased expression of senescence biomarkers (e.g., p16Ink4a and p21Cip1) and 
resistance to apoptosis(1,11). Senescent cells also develop the senescence-associated secretory 
phenotype (SASP), consisting of pro-inflammatory cytokines, chemokines, and matrix-degrading 
proteins that have deleterious paracrine and systemic effects(12-16). Further, because senescent 
cells accumulate in multiple tissues in temporal and spatial synchrony with age-associated 
functional decline in animals and humans(5-7,17-24), they have been hypothesized to drive the 
deterioration underlying numerous chronic diseases(1). Indeed, even a relatively low abundance 
of senescent cells (e.g., ~10-15% in old primates(23)) is sufficient to cause tissue dysfunction. 
Therefore, senescent cells represent promising therapeutic targets to delay or alleviate functional 
deficits and chronic disorders of aging, including age-related bone loss. 
Consistent with this, eliminating even a relatively small proportion (~30%) of senescent cells 
using a “suicide” transgene, INK-ATTAC (p16Ink4a-linked apoptosis through targeted activation 
of caspase(25)), that permits inducible elimination of p16Ink4a-expressing senescent cells upon 
administration of a drug (AP20187), extends healthspan and prevents the development of 
multiple age-related morbidities in both progeroid and naturally aged mice(12,25-27). As an 
alternative to this genetic approach and one applicable to humans, the Kirkland group at Mayo 
Clinic exploited the dependence of senescent cells on specific pro-survival pathways and 
identified that the combination of Dasatinib (D; an FDA-approved tyrosine kinase inhibitor 
[TKI] in clinical use for treating hematologic disorders(28,29)) plus Quercetin (Q; a natural 
flavanol present in many fruits and vegetables(30) that targets BCL-2, insulin/IGF-1, and HIF-1α 
networks(31-33)) [D+Q], has in vitro senolytic activity by killing senescent cells without affecting 
proliferating or quiescent, differentiated cells(31). 
To test this approach in vivo, work by our group as well as independent laboratories using mouse 
models of accelerated or natural aging has demonstrated that intermittent D+Q therapy delays the 
onset of several age-related disorders(7,26,27,31,34-41). For example, intermittent D+Q therapy 
extends healthspan in mice by preventing age-related bone loss(27) and improving several aspects 
of physical dysfunction/frailty(39) as well as atherosclerosis(26), idiopathic pulmonary fibrosis 
(IPF)(36), and tau-mediated neurodegenerative disease(38). Intermittent D+Q therapy also extended 
maximal lifespan in old mice as post-treatment survival was significantly increased by 36%(39). 
More recent findings(34,40) as well as our unpublished data have established that Fisetin (F; a 
flavonol with similar chemical structure to Q(42,43)) also has potent senolytic activity both in 
vitro(34,40) and in vivo(40), with beneficial effects on multiple tissues in both progeroid and young 
wild-type (WT) mice “aged” by senescent cell transplantation. Thus, intermittently treating aged 

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mice with D+Q or F enhances healthspan(7,26,27,31,34-41), which sets the stage for clinical studies 
testing intermittent senolytic therapy in humans as proposed in this randomized controlled trial 
(RCT). 
This study will include normal, elderly postmenopausal women, aged ≥60years, with increased 
senescent cell abundance in peripheral blood T-lymphocytes (PBTL) at screening by T-cell 
assay. 
1.2 
Investigational Agent 
 
An investigational product is defined as a pharmaceutical form of an active substance being 
tested or used as a reference in a clinical study, including products already with a marketing 
authorization but used or assembled (formulated or packaged) differently than the authorized 
form, or used for an unauthorized indication, or when used to gain further information about the 
authorized form. The investigational products will be stored in a secure area according to local 
regulations. Investigators will have responsibility for ensuring that investigational product is only 
dispensed to study subjects. The investigational products (Dasatinib plus Quercetin) will be 
dispensed only by authorized personnel according to regulations that apply at Mayo, Rochester. 
In this protocol, investigational products are Dasatinib plus Quercetin. 
 
 
1.2.1 Sprycel® (Dasatinib) 
 
Sprycel® (Dasatinib) is an FDA approved product (NDA #021986). SPRYCEL is a kinase 
inhibitor indicated for the treatment of:  
• Newly diagnosed adults with Philadelphia chromosome-positive (Ph+) chronic myeloid 
leukemia (CML) in chronic phase. The trial is ongoing and further data will be required 
to determine long-term outcome.  
• Adults with chronic, accelerated, or myeloid or lymphoid blast phase Ph+ CML with 
resistance or intolerance to prior therapy including imatinib.  
• Adults with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) 
with resistance or intolerance to prior therapy.  
 
Commercially available Dasatinib will be purchased for the purposes of this trial. Dasatinib will 
be supplied as 100 mg tablet white to off-white, biconvex, oval, film- coated with “BMS 100” 
debossed on one side and “852” on the other side. This will be an open label study. 
 
 
1.2.2 Quercetin 
 
Quercetin is a flavonoid present in many fruits, vegetables, and grains and is also used as an 
ingredient in supplements, beverages, or various types of foods. Typical dietary intakes are 
between 5 mg and 40 mg per day, but intakes of 200-500 mg/day are possible with high 
consumption of fruits and vegetables, especially when the peel is consumed. Quercetin has been 
safely used in amounts up to 500 mg twice daily for up to 8 weeks. To date, no reports of 
significant toxicity have been reported, and current evidence from multiple in vitro studies 
demonstrates safety of this product as a food additive. Our anticipation of any risks of toxicity in 

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elderly postmenopausal women who will receive this drug intermittently for only three 
consecutive days at a time (five total three-day dosing regimens) is <1%. Quercetin phytosome 
dehydrate capsules equating to 1,000 mg total daily dosage will be administered for three 
consecutive days orally to the elderly postmenopausal women randomized to the D+Q group. 
They will receive two capsules in the morning, two capsules in the evening.  This will be 
obtained commercially from the Thorne research company.  
 
Quercetin will be supplied as quercetin phytosome (sophora japonica concentrate (leaf) / 
phosphatidylcholine complex from Sunflower) 250 mg by Thorne Research. Quercetin 
Phytosome is a “00” hypromellose (vegetarian cellulose) capsule filled with a pale, yellow 
powder containing 250 mg quercetin phytosome. Microcrystalline cellulose, leucine, and silicon 
dioxide are added as manufacturing aids. This will be an open label study. 
1.3 
Preclinical Data 
Discovery of senolytics by targeting the Achilles’ heel of senescent cells: from transcriptome to 
drug identification. The class of small molecule drugs that can selectively induce death (or 
eliminate) senescent cells are known as “senolytic(s)” – i.e., from the word “senescence” and 
“lytic” – destroying. Our colleagues in the Kirkland laboratory and Mayo Clinic Kogod Center 
on Aging discovered senolytics including the combination of Dasatinib plus Quercetin (D+Q) 
and Fisetin (F)(31,34). Dasatinib is a tyrosine kinase inhibitor (TKI) that is currently used for 
certain cancer treatments and has been approved for treatment of chronic myeloid leukemia 
(CML). Dasatinib initially induces senescence followed by necrosis in cancer cells. Recent work 
evaluating the effects of Dasatinib against thyroid cancer cell lines in vitro and in a xenograft 
model in vivo, demonstrated that Dasatinib-treated cells (BHP2-7 and Cal62) exhibited a 
characteristic clumped appearance upon biopsy and most of these cells were classified as 
senescent using a common technique for assessing cellular senescence – i.e., the senescence-
associated β-Galactosidase (SA-β-Gal) assay. This work further showed that the average number 
of senescent cells per high powered field were: BHP2-7, control 0/hpf; BHP2-7, Dasatinib 
20/hpf; Cal62, control 0/hpf; Cal62 Dasatinib 25/hpf. In addition, when these senescent cells 
were followed, they displayed necrotic features after 10 days of Dasatinib treatment. 
 

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Senescent cell eradication (or reduction) 
by activating a drug-inducible “suicide” 
gene (i.e., via p16INK4a-linked apoptosis 
through targeted activation of caspase 
[INK-ATTAC]) can delay multiple age-
related phenotypes in both genetically 
modified progeroid mice and natural, 
chronologically aged mice(25,57). In 
subsequent studies, the Kirkland 
laboratory at Mayo Clinic screened 46 
agents/drugs that could theoretically 
induce apoptosis preferentially of 
senescent cells in vitro. They then 
evaluated these agents and different 
combinations to determine the specific 
subset of agents that were most effective 
at eliminating senescent cells(31). Dasatinib 
preferentially reduced viability and caused cell death in senescent human preadipocytes (also 
known as mesenchymal stem cells [MSCs]), was much less effective on senescent human 
umbilical vein cells (HUVECs) (Fig. 1A). After three days of exposure, proliferating 
preadipocytes increased by 2-5fold in number compared to day 0 in the presence of Dasatinib. 
The viability of non-dividing senescent preadipocytes from the same human subjects decreased 
by 30-40% in the presence of Dasatinib, thus demonstrating a selective reduction. 
 
Another senolytic agent our colleagues in the Kirkland laboratory discovered was Quercetin (Q), 
a naturally occurring flavonoid known to inhibit PI3Kinase, 
other kinases, and mTOR pathways. In contrast to Dasatinib, at 
low concentrations Quercetin reduced the viability and caused 
cell death of senescent HUVECs to a greater extent than 
proliferating cells, but was less effective on preadipocytes (Fig. 
1B). The combination of D+Q displayed selective killing of both 
senescent preadipocytes and endothelial cells (Fig. 2). By day 3, 
the viability of non-dividing senescent preadipocytes exposed to 
D+Q was reduced by ~70% compared to day 0, while non-
senescent, proliferating cells had increased by 2-4 fold, 
establishing that the combination of D+Q selectively targets a 
broader range of senescent cell types then either agent alone, 
hence forming the basis of our hypothesis in our study in mice 
(see below). In a previous study published in collaboration with 
the Kirkland laboratory(31), it was shown that the combination of 
D+Q not only decreased the senescent cell burden in mice in 
vivo, but also improved functional improvements in running 
endurance in younger mice that were “aged” by administration of doxorubicin, a 
chemotherapeutic drug known to cause DNA damage and induce cellular senescence. In 
addition, it was demonstrated that intermittent D+Q administration alleviated age-related cardiac 
and vascular dysfunction, improved gait in mice disabled exposing a limb to ionizing radiation, 
Fig. 1. Effects of (A) Dasatinib and (B) Quercetin 
on preadipocytes and HUVECs. 
Fig. 2. Effects of Dasatinib 
plus Quercetin (D+Q) on 
preadipocytes and HUVECs. 

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and delayed frailty, neurological dysfunction, and osteoporosis in mice with an accelerated 
aging syndrome(31). These findings were later further supported by studies in our laboratory more 
specifically examining the roles of senescent cells in mediating age-related bone loss (detailed 
below). 
  
Identifying senescent cells in bone and the causal role of cellular senescence in age-related 
bone loss. Our group recently found(7) in mice that with aging, a subset of cells of various 
lineages within the bone microenvironment become senescent and that some of these senescent 
cell populations develop a SASP. Importantly, we found similar results in bone biopsies from 
older vs younger women(7), establishing that senescent cells are present at the time and location 
of age-related bone loss in 
humans, as they are in mice. 
To establish causality(27), we 
next used genetic (INK-
ATTAC(25)) and 
pharmacological (D+Q(31)) 
approaches to target senescent 
cells and assessed their impact 
on age-related bone loss in old 
mice. As detailed in our 
publication(27) and as 
summarized in our recent 
reviews(58,59), the skeletal 
effects of intermittent D+Q 
therapy in old WT mice (Fig. 
3 A-F) were virtually identical to those observed in the old INK-ATTAC mice treated with 
AP20187 (a synthetic drug that induces suicide transgene activation(25)), demonstrating that 
senescent cell clearance via either genetic or senolytic approaches prevents age-related bone loss. 
By histomorphometry, we found that targeting senescent cells using genetic or senolytic 
approaches reduced bone resorption (Fig. 3D). Importantly, despite lower bone resorption, bone 
formation was either maintained (trabecular sites) or was higher (cortical sites; Fig. 3 E, F) 
following senescent cell clearance(27). Thus, from a therapeutic perspective, targeting senescent 
cells offers substantial advantages over conventional anti-resorptive therapies(58) as the latter 
inhibit/eliminate osteoclasts to reduce bone resorption, with a concomitant reduction in bone 
formation due to coupling. By contrast, senolytic therapy reduces the senescent cell burden, 
which in turn suppresses bone resorption with either increased (cortical bone) or maintained 
(trabecular bone) bone formation, leading to positive “bone balance”(27). The studies proposed in 
this application are aimed at translating these novel findings in mice to humans. 
Fisetin is a senolytic that extends health- and lifespan and rescues senescent cell-induced bone 
loss. As noted earlier, Fisetin (F – 3,3’,4’,7-tetrahydroxyflavone) is a member of the flavonoid 
family of naturally occurring polyphenolic compounds present in many fruits and vegetables with 
high concentrations in strawberries (160 µg/g)(42,43). To date, no adverse effects of F have been 
reported, even when taken at high doses(60). The hydrophobic nature of F allows it to penetrate cell 
membranes and accumulate in cells to exert its effects(49,52). In vitro studies in either senescent 
primary human umbilical vein endothelial cells or senescent primary murine embryonic fibroblasts 
demonstrate that F significantly reduced viability (causing apoptosis of 30-70% of senescent cells) 
0 .0 0
0 .0 5
0 .1 0
0 .1 5
C t.T h . (m m )
0 .0
0 .5
1 .0
1 .5
B F R  / B S
 (m c m 3/m cm 2/d )
0
5
1 0
1 5
N .O b /B .P m  (/m m )
0
5
1 0
1 5
N .O c /B .P m  (/m m )
0
5
1 0
1 5
2 0
B V /T V  (% )
0
1 5
3 0
4 5
6 0
7 5
F a ilu re  L o a d  (N )
**
F
**
D
E
**
C
*
B
*
A
*
Vehicle
D+Q
Fig. 3. D+Q (n=10) prevents age-related bone loss in old WT mice 
vs vehicle (n=13). (A-C) µCT-derived spine bone volume fraction 
(BV/TV), femur cortical thickness (Ct.Th.), and femur failure load 
(strength). (D-F) Histomorphometry (n=8/group) at the femoral 
endocortical surface: osteoclast #’s/bone perimeter (N.Oc/B.Pm), 
osteoblast #’s/bone perimeter (N.Ob/B.Pm), and bone formation 
rate/bone surface (BFR/BS). Mean ± SEM. *p<0.05; **p<0.01. 

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and senescence biomarkers relative to vehicle(34,40). Notably, eliminating ~30% of senescent cells 
in INK-ATTAC mice was sufficient to  alleviate multiple age-related conditions(25). In vivo studies 
in both naturally aged WT mice and Ercc1-/∆ mice, which model human XFE progeria and display 
several features resembling accelerated aging within a lifespan of 6 months(61), demonstrated that 
F significantly reduced senescence biomarkers, while late-life intervention with F in old WT mice 
significantly extended lifespan(40). Similar findings were observed in human adipose tissue 
explants treated with F vs vehicle(40). F also extended healthspan measured by a composite frailty 
score in the Ercc1-/∆ mouse model of accelerated aging(40), thus establishing beneficial effects of F 
on physical function and aspects of frailty. Finally, our more recent unpublished data in young 
adult (4-month) WT mice transplanted with either relatively small numbers (i.e., 106) of quiescent, 
non-senescent control cells or senescent cells (Fig. 4), demonstrate that senescent cell 
transplantation causes significant bone loss that is rescued by intermittent F therapy (Fig. 4 A-B).  
Collectively, these data in 
mice establish that reducing 
the senescent cell burden with 
senolytics has broad potential 
implications for osteoporosis 
and other chronic diseases of 
aging. 
Clinical Data to Date 
The senolytic drugs used in 
this study include Dasatinib 
and Quercetin. Dasatinib has 
previously been approved for 
the treatment of chronic myeloid leukemia (CML) by the FDA. This proposal outlines a new 
indication for use and will be taken orally on an intermittent schedule (starting every 28 days) with 
no-therapy periods in between dosing regimens, repeated every 28 days over 20 weeks. More 
specifically, the oral treatment administration for each dosing regimen will be Dasatinib (D; 100 
mg daily for two consecutive days) plus Quercetin (Q; 1000 mg total daily for three consecutive 
days) taken orally on an intermittent schedule (starting every 28 days) with no-therapy periods in 
between dosing regimens, repeated every 28 days over 20 weeks, resulting in five total dosing 
regimens throughout the entire intervention. 
 
To our knowledge, there are no published studies utilizing Dasatinib or Quercetin or the 
combination of both drugs in the population of normal elderly postmenopausal women, 
independent of a diagnosis of CML. Dasatinib is a TKI used as an oral daily treatment for CML 
with fewer side effects and improved survival compared to older treatment agents for this 
disease(62). Dasatinib is generally well-tolerated with the more common adverse effects consisting 
of rash, abdominal pain, diarrhea, nausea, myelosuppression, fluid retention, and headache. 
 
There has, however, been a recent publication by Yilmaz et al.(63) in which a historical cohort 
review was conducted on 468 newly diagnosed patients with chronic-phase chronic myeloid 
leukemia at MD Anderson who were treated long-term with TKIs (Imatinib, Dasatinib, and 
Nilotinib). The primary goal was to examine the incidence of changes in kidney function over 
time. Among this study cohort, 99 of 468 patients received Dasatinib dosed at 100 mg daily or 50 
Fig. 4. After two months (2-mo), senescent (SEN) cell transplantation 
in young adult (4-mo-old) WT mice causes significant bone loss [(A) 
lower spine BV/TV and (B) trabecular thickness (Tb.Th.)] vs control 
(CON) cell transplantation that is rescued by intermittent (bi-weekly) 
fisetin (F) therapy vs vehicle (V). Mean ± SEM. *p<0.05; **p<0.01. 
0 .0 0
0 .0 2
0 .0 4
0 .0 6
0 .0 8
0 .1 0
T b .T h . (m m )
0
7
1 4
2 1
2 8
3 5
B V /T V  (% )
CON
+ V
SEN 
+ V
2-mo
Skeletal 
Measurements
SEN 
+ F
*
*
*
**
A
B
Cell Transplantation
n = 15-17 / group

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mg twice daily for more than 3 months and were followed for a median of 39 months. Among the 
Dasatinib treated patients, only one of 99 patients developed acute kidney injury defined as a rise 
in serum creatinine of 0.3 mg/dL. Among the 93 Dasatinib-treated patients with normal baseline 
kidney function, the mean estimated glomerular filtration rate (eGFR) initially declined and then 
stabilized after four years. Among the six Dasatinib-treated patients with CKD (eGFR <60 
mL/min) at baseline, there were no significant changes in eGFR over time within this relatively 
small group. The overall findings were that long-term Dasatinib-treated patients with normal 
baseline kidney function did experience a decline in eGFR but no association was found with 
development of acute kidney injury. Additionally, those with CKD at baseline did not experience 
any further decline in eGFR (for either of the tyrosine kinase inhibitors studied). 
 
Dasatinib (D) has rarely been associated with acute kidney injury among patients treated for 
leukemia. The injury may occur in the context of tumor lysis syndrome which is of minimal to 
no risk for CKD patients who lack the underlying hematological disease process. Among patients 
treated for leukemia, available case reports also describe a variety of kidney lesions including: 
acute tubular necrosis from adverse effects such as gastroenteritis and nephrotic syndrome from 
thrombotic microangiopathy occurring in patients receiving Dasatinib therapy(64-66). These events 
generally resolve with interruption of drug therapy and conservative management. Based on a 
case report and review by Wallace et al.(66), a possible mechanism for Dasatinib-induced kidney 
injury is through inhibition of the vascular endothelial growth factor (VEGF) signaling pathway, 
a specific tyrosine kinase signaling pathway. However, there are no package insert recommended 
dosing adjustments for either kidney disease or reductions in kidney function for patients 
receiving Dasatinib therapy. 
 
Quercetin (Q) is a natural-occurring flavonoid known to inhibit PI3Kinase, other kinases, and 
mTOR pathways. Quercetin is present in many fruits, vegetables, and grains and is also used as 
an ingredient in supplements, beverages, or various types of foods. Quercetin is a supplement 
and not FDA approved for any indication. The primary contraindication/warning for this drug is 
a hypersensitivity to Quercetin. Adverse effects associated with quercetin include emesis, 
dyspnea, and nephrotoxicity or kidney injury. Kidney injury from this drug has not been 
substantiated in recent reports and overall appears safe in clinical application(67), and several 
studies involve use of Quercetin for its anti-oxidative and anti-apoptotic effects in animal models 
of kidney disease, including diabetic nephropathy (DN). In fact, there are numerous preclinical 
studies in acute kidney injury or DN suggesting that Quercetin may reduce oxidative stress and 
improve kidney function(68-71). 
 
1.4 
Dose Rationale  
Reducing the senescent cell burden with senolytics extends health- and lifespan in old 
mice(7,26,27,31,34-41), and prevents age-related bone loss(7,27). The senolytics proposed in this 
application were discovered using a hypothesis-driven approach whereby senolytics selectively 
induce senescent cell apoptosis by transiently disabling the senescent cell anti-apoptotic 
pathways (SCAPs)(32,77) that defend senescent cells against their own pro-apoptotic 
environment(31). However, not all senescent cells are the same: they can originate from different 
cell types, express unique SASPs(78-80), and use different mechanisms to resist apoptosis(32,77). 
Intermittent D+Q or F therapy in old mice or young mice “aged” by senescent cell 

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transplantation alleviates a range of age-related chronic conditions, including osteoporosis(27). 
Because senescent cells do not divide(1) and take at least weeks to re-accumulate(81) and because 
senolytics do not need to be continuously present to occupy a receptor or affect an enzyme(32,77), 
senolytic combinations, such as D+Q(31), and newer senolytics such as F(40), can be repurposed 
for immediate translation to humans. This premise is supported by a first-in-human pilot study of 
intermittent D+Q therapy(82), showing that senolytics alleviated aspects of physical dysfunction 
in patients with IPF, a progressive, fatal cellular senescence-associated disease(36,83,84). This study 
provides “proof-of-concept” feasibility for targeting senescent cells in humans with senolytics to 
alleviate aspects of age-related diseases, and supports additional studies evaluating senolytics for 
other chronic diseases of aging, such as osteoporosis. 
Dasatinib plus Quercetin (D+Q): D is an FDA-approved TKI used clinically to treat 
hematologic disorders with a long-standing, acceptable safety profile(28,29). The oral dose of D 
(100 mg/d) selected is based on the FDA-approved dose for chronic therapy as effective for 
inducing apoptosis in human cancer cells. Follow-up analysis (5-years) from the phase III 
Dasatinib versus Imatinib Study in Treatment-Naïve Chronic Myeloid Leukemia (CML) Patients 
trial(85), demonstrated long-term efficacy and safety of oral D (100 mg/d). In addition, in the TKI 
era (e.g., with daily D therapy) patients with CML have lifespans nearly as long as the general 
population(86). In contrast to the daily D regimen these patients receive, D therapy in our study 
will be administered intermittently (see below). The oral dose of Q (a natural product present in 
many fruits and vegetables(30)) selected is 1000 mg/d (250 mg capsules x 4/d). Importantly, in the 
first-in-human pilot study of D+Q in patients with IPF(82), these doses of D+Q for 3 consecutive 
days over 3 consecutive weeks (9 total participant administered dosing days) at two separate U.S. 
clinical sites were well tolerated without notable changes in clinical chemistries or renal/hepatic 
function. Also, in a more recent study, we found that these doses of D+Q for 3 consecutive days 
demonstrated clearance of senescent cells in the skin and adipose tissue (87). Based on pre-clinical 
models, we believe that 2 doses of D along with 3 doses of Q will be equally effective, but a 
single dose of D may not be; as such, we propose to use daily doses of D x2d and daily doses of 
Q x3d in our study.  In our study, the D and Q doses will be the same, but the interval between 
dosing regimens will be longer (26 [Dasatinib] or 25 [Quercetin] days off senolytic therapy 
between dosing regimens) vs the first-in-human study in IPF patients (4 days off senolytic 
therapy between dosing regimens)(82). Further, the study duration we propose is 20 total weeks to 
include a total of 5 participant dosing days administered intermittently (every 4 weeks, see 
Section 3, Study Design). More specifically, the two-consecutive day dosing days for Dasatinib 
will be at Baseline (Day 1, 2), start of Week 5 (Day 29, 30), start of Week 9 (Day 57, 58), start of 
Week 13 (Day 85, 86), and start of Week 17 (Day 113, 114) for a total of ten days of Dasatinib 
over the course of the 20-week study. More specifically, the three-consecutive day dosing days 
for Quercetin will be at Baseline (Days 1-3), start of Week 5 (Days 29-31), start of Week 9 
(Days 57-59), start of Week 13 (Days 85-87), and start of Week 17 (Days 113-115) for a total of 
15 days of Quercetin over the course of the 20-week study. 
1.5 
Risks and Benefits 
Based on the above (see Section 1.4, Dose Rationale), our anticipation of any toxicity risk in 
subjects receiving these compounds (D+Q) for only two or three consecutive days followed by at 
least 25 days of no therapy, every four weeks over the course of 20 weeks, is <1%. Therefore, 
the potential benefits of intermittent senolytic administration to target senescent cells and thereby 
alleviate multiple functional consequences of age-related diseases in humans, as in 

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mice(7,26,27,31,34-41), is considerable. Further, although not yet established, the senolytic proposed 
here and next generation senolytics may have a better safety profile than existing pharmacologic 
options for osteoporosis prevention and treatment(88). The studies proposed herein will be pivotal 
to this end. In addition, the risks of drawing blood include pain, bruising, lightheadedness, and/or 
fainting, or rarely, infection at the site of the needle stick.  Study subjects will be exposed to low 
doses X-ray radiation during the dual-energy X-ray absorptiometry (DXA) and high-resolution 
peripheral quantitative computed tomography (HR-pQCT) scans. The amount of radiation has a 
low risk of harmful effects. Based upon the review of the published literature and our pre-clinical 
as well as clinical data, we do not anticipate the occurrence of serious adverse events with any of 
the procedures or prescribed drug dosing regimen in this study. We have identified a variety of 
drug-drug interactions for which we established exclusion criteria or modification plans to 
minimize associated risks. Therefore, the overall procedure risks and any risks of using either 
drug regimen are likely minimal compared to the anticipated benefits and the knowledge that 
may be gained from this clinical investigation. 
2 Study Objectives 
 
Primary Objective 
 
In a cohort of 60 older postmenopausal women, aged ≥60years, with a high senescent cell burden 
(as these individuals are most likely to benefit from pharmacological interventions that eliminate 
senescent cells – i.e., senolytics), we will test the efficacy of intermittent senolytic therapy 
(D+Q) as compared to non-interventional therapy on improving serum bone resorption markers 
over the course of 20 weeks. 
 
 
Secondary Objective 
 
In the same cohort of 60 older postmenopausal women, aged ≥60years, with a high senescent 
cell burden, we will further assess the impact of senolytics (D+Q) as compared to non- 
interventional therapy on reducing systemic surrogate markers of senescent cell abundance and 
circulating senescence-associated secretory phenotype (SASP) factors. 
 

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3 Study Design 
 
 
 
 
3.1 
General Description 
We will conduct a 20-week, open label, RCT comparing D+Q to the control group for improving 
bone turnover markers and skeletal parameters and 
in reducing systemic surrogate markers of 
senescent cell abundance and the SASP in 80 older 
postmenopausal women with an underlying 
senescent cell burden based on p16INK4a mRNA 
expression in PBTL (>95th percentile of same sex 
young controls; cut-off is shown in Fig. 6). All 
studies will be performed at the Mayo Clinic 
outpatient Clinic Research Unit (CRU) following 
IRB approval. At screening, the nurse study 
coordinator will explain the study, including 
potential benefits and risks to volunteer subjects. 
After obtaining informed written consent and 
following comprehensive screening studies 
(including blood draw after 12-hr overnight fast for 
the p16INK4a PBTL assay and clinical chemistries), subjects who meet eligibility criteria (as 
detailed below) and agree to take the assigned therapy for the entire study duration, will be 
randomized to one of two groups: i) untreated control group; ii) D (100 mg/d, for two days,  
Sprycel, Bristol Myers Squibb) plus Q (1000 mg/d; 250 mg capsules x 4/d Quercetin Phytosome, 
Thorne Research). As detailed above, D therapy will be taken orally on an intermittent schedule 
(every 27 days) with no-therapy periods in between doses, repeated every 28 days over 20 
weeks, resulting in ten total dosing days throughout the entire intervention, whereas Q therapy 
Fig. 6. p16
INK4a mRNA in PBTL as a biomarker 
to permit detection of in vivo human cellular 
senescence. Dashed line = 95th percentile of 
younger women. Mean ± SEM. ***p<0.001. 
0
2 1 0 - 5
4 1 0 - 5
6 1 0 - 5
p16 IN K 4 a m R N A
(N o rm a liz e d  E x p re s s io n )
Potentially 
eligible 
older 
subjects w/ 
underlying 
cellular 
senescence
***
Older women (n=20)
Younger women (n=20)
 

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will be taken orally on an intermittent schedule (every 25 days) with no-therapy periods in 
between doses, repeated every 28 days over 20 weeks, resulting in 15 total dosing days 
throughout the entire intervention. Our proposed intermittent dosing regimen is based on the 
following: i) effects of D+Q on various senescent cell types in culture(31,34,40); ii) our preclinical 
studies in which senolytics were administered monthly to naturally aged old WT mice (Fig. 3); 
iii) senescent cell clearance from freshly isolated human tissue explants treated with D+Q(39) or 
F(40); iv) peak concentrations and the short elimination half-lives (<12 hrs.) of these senolytics in 
humans(89,90); and v) dose escalation studies using these senolytics in old mice and old Rhesus 
monkeys. Block randomization will be performed by our statistician, Ms. Elizabeth Atkinson, 
M.S., to ensure balanced group assignment as the study proceeds. The first dose of each dosing 
regimen will be administered onsite by the nurse study coordinator who will provide eligible, 
randomized subjects with a screw cap bottle(s) from the Mayo Clinic Research Pharmacy 
containing their group allocated therapy (D+Q).  Throughout the study, all subjects will be 
instructed to take 1000 IU of Vitamin D (supplied to them) and will be counseled to obtain total 
calcium intake of ~1200 mg/d.  At each visit, a fasting morning blood draw will be obtained. 
Subjects will be given a blank notebook for recording AEs that the study team will review and 
document at each visit. The primary study endpoints will be percent changes in serum bone 
turnover markers (amino-terminal propeptide of type I collagen [P1NP] and C-terminal 
telopeptide of type I collagen [CTX]) from baseline over 20 weeks; changes in these markers 
over intermediate time points (weeks 4, 8, 12, 16) and changes in the P1NP/CTX ratio will be 
considered secondary endpoints. Additional secondary endpoints will be patient reported quality 
of life, safety and tolerability and changes in the following from baseline to study endpoint: i) 
bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) at the lumbar spine, 
hip (total and femoral neck [FN]), and radius (total and ultra-distal); ii) high-resolution 
peripheral quantitative computed tomography (HRpQCT)-derived trabecular/cortical bone 
parameters and micro-finite element analysis (µFEA)-derived bone strength at the radius and 
tibia; iii) p16INK4a mRNA expression in PBTL; and iv) plasma SASP factors. 
3.2 
Number of Subjects 
60 elderly postmenopausal women, aged ≥60 years, with increased senescent cell abundance in 
blood will proceed to randomization. We will screen 240 elderly postmenopausal women, aged 
≥60years, as we anticipate ~50% will have eligible PBTL p16INK4a mRNA expression assays. 
Note that 14 subjects have already completed the F group, but no additional subjects will be 
enrolled into that group per approval from NIH and our NIH DSMB (see documentation). Data 
from these 14 subjects will be used as pilot data to design future studies using F, depending on 
the outcome from D+Q. 
3.3 
Duration of Participation 
Participants will remain in the study for 20 weeks post accrual. In addition, a follow-up phone 
call will be performed at 1 and 2-year post-study to inquire about any new malignancies that may 
have occurred since completing the study. 
3.4 
Primary Study Endpoints 
The primary study endpoints will be percent changes in the serum bone resorption marker (C-
terminal telopeptide of type I collagen [CTX]) from baseline over 20 weeks. 

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3.5 
Secondary Study Endpoints 
Changes in the bone formation marker, amino-terminal propeptid of type I collagen [P1NP] and 
CTX markers over intermediate time points (weeks 4, 8, 12, 16) and changes in the P1NP/CTX 
ratio will be considered secondary endpoints. Additional secondary endpoints will be patient 
reported quality of life, safety and tolerability and changes in the following from baseline to study 
endpoint: i) bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) at the 
lumbar spine, hip (total and femoral neck [FN]), and radius (total and ultra-distal); Due to the 
recent availability of trabecular bone score (TBS) software on the CRTU iDXA as an alternative 
method for assessment of bone quality from DXA images, we will explore how vertebral TBS 
compares to the cortical measures (BMSi and cortical porosity) in secondary analyses.  Note that 
TBS requires no additional DXA imaging; it is performed on L1-L4 AP spine scans which have 
already been performed. ii) high-resolution peripheral quantitative computed tomography 
(HRpQCT)-derived trabecular/cortical bone parameters and micro-finite element analysis 
(µFEA)-derived bone strength at the proximal and distal radius and tibia; iii) p16INK4a mRNA 
expression in PBTL; and iv) plasma SASP factors. 
3.6 
Primary Safety Endpoints 
Safety assessments are undertaken with the measurement of safety laboratory tests and 
procedures, vital signs, and recording of adverse events. In addition, follow-up phone calls will 
be performed at 1 and 2-year post-study completion to inquire about new malignancies.  The 
following safety labs will be monitored and limits on these are as noted, which would result in 
withdrawal of the subject from the study:  ECG (QTc > 470 msec); CBC (PI’s assessment of 
clinically meaningful changes in blood counts; if ambiguous, this will be reviewed with an 
independent clinician [Dr. Matthew Drake]); AST (>2x above the normal limit); cystatin C (> 
50% increase or reduction of eGFR to below 30 ml/min/1.73 m2). 
3.7 
Identification of Source Data 
Source data will be obtained from the electronic medical record and documented in study 
specific case report forms (paper and/or electronic data capture). No information in source 
documents about the identity of the subjects will be disclosed. 
4 
 Subject Selection Enrollment and Withdrawal 
4.1 
Inclusion Criteria 
We will randomize 60 eligible older postmenopausal women aged ≥60years to one of two groups: 
untreated control group or to D+Q; Inclusion criteria are as follows: 
• Able and willing to provide informed consent. 
• Normal postmenopausal women. 
• Aged ≥60years. 
4.2 
Exclusion Criteria 
• Subjects who are Type II diabetic and on insulin due to a risk of hypoglycemia. 
• Hemoglobin A1c ≥8.0% at screening 
• Abnormal screening labs (see below): Calcium >10.1 mg/dL, Phosphorus >4.7 mg/dL, 
Thyroid stimulating hormone (TSH) level <0.3mU/L, Fasting blood glucose >200 mg/dL.   

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• Presence of significant liver (with total bilirubin >2X upper normal limit, AST >2xupper 
normal limit, alkaline phosphatase >2x upper normal limit) or kidney disease (see eGFR 
below). If any elevations were to be noted (>2x the normal level), the study participant 
would stop treatment and have levels re-drawn in a month, per the clinical judgement of 
the investigator.  
• If any of the laboratory blood work drawn at the study visits return with lab values 
outside of the “normal limits” or show a significant change from a previous value, a 
repeat blood draw would be done before the subject is excluded. 
• Presence of a clinical diagnosis of heart failure 
• Known active malignancy (including myeloma) 
• Current diagnosis of Malabsorption or currently undergoing treatment for malabsorption 
disease 
• gastric bypass / reduction 
• Hyperthyroidism 
• Acromegaly 
• Cushing’s syndrome 
• Hypopituitarism 
• Subjects with a fracture within the past six months 
• Undergoing treatment with any medications that affect bone turnover, including the 
following: 
o adrenocorticosteroids (> 3 months at any time or > 10 days within the previous 
yr., except for use of topical steroid creams or gels or inhaled steroids), 
anticonvulsant therapy (within the previous year, include only those taking 
Carbamazepine, Phenobarbital and Phenytoin), 
o bisphosphonates (within the past 3 yrs., including injections or infusions); oral use 
permitted if taken for less than 1 month, 
o denosumab, 
o estrogen (E) therapy or treatment with a selective E receptor modulator, except 
for vaginal estrogen cream use, or teriparatide, abaloparatide, romosozumab (>1 
month within the past yr.) 
 
• QTc >450 msec 
• Current diagnosis of hypo- or hyperparathyroidism or currently undergoing treatment for 
the disease 
• Inability to provide consent 
• Inability to tolerate oral medication 
• eGFR<30 ml/min/1.73 m2 (using the cystatin C blood levels for analysis)  
• Subjects on therapeutic doses of anti-coagulants (e.g. warfarin, heparin, low molecular 
weight heparin, factor Xa inhibitors, etc) 

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• Subjects with hypovitaminosis D (25-hydroxyvitamin D [25(OH)D] <20 ng/ml, whose 
level does not improve above 20 ng/ml after two courses of 4-week treatment of 50,000 
IU/d of Vitamin D.  They will be referred to their primary provider should this occur. 
• Subjects taking anti-arrhythmic medications known to cause QTc prolongation 
• Subjects taking potentially senolytic agents within the last 6 months: Fisetin, Quercetin, 
Luteolin, Dasatinib, Piperlongumine, or Navitoclax 
• Subjects currently taking drugs that induce cellular senescence: alkylating agents, 
anthracyclines, platins, other chemotherapy 
• Subjects taking H2 antagonists, unless randomized to the control group 
• Tyrosine kinase inhibitor therapy 
• Subjects not having a PBTL p16INK4a  mRNA expression level >95 percentile of young 
female controls (this cut-off is depicted by the dotted line in Fig. 6) 
• Known hypersensitivity or allergy to Dasatinib or Quercetin 
• Subjects taking the following antimicrobial agents: Aminoglycosides, Azole antifungals 
(fluconazole, miconazole, voriconazole, itraconazole), Macrolides (clarithromycin, 
erythromycin), Antivirals (nelfinavir, indinavir, saquinavir, ritonavir, 
elbasvir/grazoprevir), Rifampin 
• Subjects taking medications that are sensitive to substrates or substrates with a narrow 
therapeutic range for CYP3A4, CYP2C8, CYP2C9, or CYP2D6 or strong inhibitors or 
inducers of CYP3A4 (e.g., cyclosporine, tacrolimus or sirolimus). If antifungals are 
necessary from an infectious disease perspective, then they will be allowed only if the 
levels are therapeutic.  
• Subjects taking strong inhibitors of CYP3A4  
• Subjects on antiplatelet agents (Clopidogrel [Plavix]; Dipyridamole + Aspirin 
[Aggrenox]; Ticagrelor [Brilinta]; Prasugrel [Effient]; Ticlopidine [Ticlid] or Other) who 
are unable or unwilling to reduce or hold therapy prior to and during the study drug 
dosing periods. Subjects may continue their previous regimen between study drug dosing 
periods.  
• Subjects on quinolone antibiotic therapy for treatment or for prevention of infections 
within ten days. 
• Subjects taking proton pump inhibitors and unwilling to discontinue therapy for one week 
before and two weeks following dosing. 
• Subjects with clinically evident fluid retention 
• Subjects with evidence of right heart strain on ECG 
• Subjects with a history of pulmonary hypertension 
• Subjects with an abnormal Complete Blood Count (clinically insignificant changes would 
be acceptable based on the judgement of the investigators) 
• Presence of any condition the Investigator believes would place the subject at risk or 
would preclude the subject from successfully completing all aspects of the trial. 
• If the DXA assessment reveals a spine or femur neck T-score < -2.5, the participant will 
be advised of this. She would then be given the option of withdrawing from the study to 
immediately start an osteoporosis drug through her primary care physician or continue in 

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the study and defer osteoporosis drug treatment for the duration of the study (20 weeks). 
Given that osteoporosis is a chronic, long-term disease, the 20-week deferral would pose 
a minimal risk to the participant and she would be free to make this choice. 
 
 
Behavioral Modification: Participants will be educated about the risk of excessive caffeine usage. 
Participants will be encouraged to reduce use by 50% prior to and during the drug dosing periods. 
Due to drug-drug interaction, subjects may not clear the caffeine from their system properly/as 
usual.  Because of possible effects on the metabolism of dasatinib, subjects will also be advised 
not to consume grapefruits or grapefruit juice during their participation in the study. Patients will 
be asked to contact the study coordinator by phone if prescribed any new medication during the 
trial. 
 
Involvement of special vulnerable populations: We will not involve special vulnerable 
populations, such as fetuses, neonates, pregnant women, children, prisoners, institutionalized 
individuals, or others who may be considered vulnerable populations. 
4.3 
Subject Recruitment, Enrollment and Screening 
Recruitment sources will include: 1) Previous research study participants, 2) flyers, 3) classified 
advertising, 4) radio advertising, 5) newspaper advertisement, 6) appropriate internet/social 
media platforms 
 7) elderly population-
focused newsletters 
; Family 
Service Rochester Senior Independence; 
; Rochester Family Service-
Neighbors Helping Neighbors; 
/; Elder Network; 
), 8) Mayo clinic patients (EMR search)and 9) at off-site campuses 
(e.g., retirement communities). Permission will be obtained from each location before 
recruitment begins.  Trained clinical coordinators will contact subjects and assist in recruitment 
and carry-through of the protocol.  Description of the studies and review of the informed consent 
forms with each patient will be conducted in person directly with the clinical coordinators 
involved in the study. Written informed consent will be obtained with details of the procedures to 
be followed, the number of subjects to be included, identification of the risks and benefits, and 
alternative procedures.  Please see Inclusion and Exclusion criteria above. Documentation of 
recruitment and enrollment efforts will be maintained in a secure database. We anticipate 
enrolling 60 subjects (30 per group). All studies will be performed and samples collected in the 
Clinical Research and Trials Unit (CRTU) at Mayo, with study subjects maintaining their usual 
diet (including usual calcium intake) and Vitamin D use. Screening will include a brief history, 
physical exam, ECG, and the following fasting blood screening tests: CBC, HbA1c, serum 
calcium, creatinine, aspartate aminotransferase, 25(OH)D, fasting blood glucose, bilirubin, 
serum phosphorus, alkaline phosphatase, thyroid-stimulating hormone and cystatin C. 
 
At the initial screen (Visit 1), subjects will undergo a blood test for peripheral blood CD3+ T cell 
assay (in PBTL) for p16INK4a mRNA expression and at Baseline (Visit 2) they will undergo a test 
using an AGES reader. These tests will be analyzed in our laboratory as we have established 
criteria for determining the presence of significant cellular senescence. If there is absence of 
cellular senescence in the T cell assay, the subject will not undergo randomization. However, if 
the T cell assay does reveal cellular senescence (as depicted in Fig. 6), then they will proceed to 

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randomization. The rationale for this selection is that senolytic drugs are unlikely to be effective 
in the absence of underlying cellular senescence. 
 
4.4 
Early Withdrawal of Subjects 
4.4.1 When and How to Withdraw Subjects 
All subjects will be assessed during the three days of medication administration, and subsequent 
days of medication administration and all study visits. If a severe adverse event (SAE) occurs at 
any time during administration of the three-day drug regimen, a formal review will occur and 
subsequent patients will be enrolled one at a time using the same regimen. If three or more 
events accrue, the study will be held and either a potential dosing regimen revisited or 
discontinuation of the study protocol will occur. Other interventions will be as per the direction 
of the Food and Drug Administration and Mayo Institutional Review Board. 
Study Completion: 
For each subject in the study, the end of study will be reached when treatment and post-treatment 
safety follow-up periods have been completed. 
Subject withdrawal: 
A subject may be withdrawn from the study prior to that subject completing all study- related 
procedures. Some reasons may include: 
• Subject safety issues 
• Failure of subject to adhere to protocol requirements 
• Subject decision to withdraw from the study (withdrawal of consent) 
Withdrawn subjects may not re-enter the study. 
 
Premature withdrawal from study: 
Subjects may voluntarily withdraw from the study for any reason at any time. Subjects are 
considered withdrawn if they state an intention to withdraw further participation in all 
components of the study, die, or are lost to follow-up for any other reason. The investigator may 
withdraw a subject from the study (without regard to the subject’s consent) if they believe that 
continued participation in the study would be contrary to the best interests of the patient. 
Subjects are considered as lost to follow-up if all reasonable attempts by the investigator to 
communicate with the individual fail. The investigator will take preventive measures to avoid a 
subject being lost to follow-up (e.g., document different ways of contact such as telephone 
number, home address, e-mail address, person to be contacted in case the subject cannot be 
reached). If the subject cannot be reached, the investigator will make a reasonable effort to 
contact the subject, document all attempts and enter the loss of follow-up information into the 
Case Report Form (CRF). The following methods will be used: at least two telephone calls will 
be placed to the last available telephone number (each call on different days) and one registered 
letter will be sent by post to the last available home address. If the subject is still unreachable 
after all contact attempts listed above, he/she will be considered lost to follow-up. 
If premature withdrawal occurs for any reason, the reason for premature withdrawal from the 
study, along with who made the decision (subject, investigator) will be recorded in the CRF. 

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Reporting of Serious Adverse Events and Unanticipated Problems: 
When an adverse event has been identified, the study team will take appropriate action necessary 
to protect the study subject and then complete the Study Adverse Event Worksheet and log. The 
investigator will evaluate the event and determine the necessary follow-up and reporting 
required. 
Subject replacement: 
Subjects withdrawn from the study will be replaced by newly recruited subjects meeting 
inclusion criteria matching similar baseline characteristics (age, sex, race, and T cell assay 
senescent cell positivity). 
4.4.2 Data Collection and Follow-up for Withdrawn Subjects 
 
• For withdrawn subjects not undergoing any study intervention, no additional follow-up 
will be done. 
• For withdrawn subjects receiving intervention (Dasatinib plus Quercetin), for safety 
monitoring telephone contact will be attempted and subjects will be encouraged to return 
to complete the laboratory evaluations at the Week 20 (Day 140) visit. No follow up 
phone call needed beyond four months, except to screen for malignancies at year 1 and 2 
post-study completion. It will be highly recommended that withdrawn subjects return for 
clinical blood (CBC, creatinine, etc.) studies as part of safety monitoring. Research data 
will not be collected on subjects after they are withdrawn from the study. Any additional 
evaluation will be for subject safety only. 
• For those subjects in the intervention groups who are not tolerating the medication and 
considering withdraw, they will be given the option to continue in the study with intent to 
treat. 
5 Study Drug 
5.1 
Description 
 
Dasatinib (commercially available) will be purchased for the purposes of this trial. Dasatinib will 
be supplied as 100 mg tablet white to off-white, biconvex, oval, film- coated with “BMS 100” 
debossed on one side and “852” on the other side. 
 
Quercetin will be supplied as quercetin phytosome (sophora japonica concentrate (leaf) / 
phosphatidylcholine complex from Sunflower) 250 mg by Thorne Research. This drug product 
will be dispensed through the research pharmacy in child resistant containers. Quercetin 
Phytosome is a “00” hypromellose (vegetarian cellulose) capsule filled with a pale, yellow 
powder containing 250 mg quercetin phytosome. Microcrystalline cellulose, leucine, and silicon 
dioxide are added as manufacturing aids. 
 
 

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5.2 
Treatment Regimen 
 
1) Dasatinib is a Tyrosine Kinase Inhibitor (TKI) used to treat cancer, particularly 
Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML) or Ph+ acute 
lymphoblastic leukemia (ALL). Dasatinib is currently FDA approved. Precautions 
include: cardiac adverse events, avoidance of use of H2 blockers and proton pump 
inhibitors (affects Dasatinib absorption), fluid retention, hemorrhage, myelosuppression, 
dermatologic reactions, and avoidance in pregnancy/breast feeding. The drug interferes 
with cytochrome P450 and may require drug adjustments to agents such as calcineurin 
inhibitors. Unlike standard treatment regimens for CML (lasting several months), this 
medication will only be administered intermittently. Elimination half-life is <12 hours in 
adults. Similarly, in animal models, drug clearance is observed within 72 hours. Yet, 
based on animal studies, senescent cell clearance persists for at least ~2-4 weeks. 
a. Dosing: Dasatinib in the form of Sprycel® (Bristol Myers Squibb) 100 mg orally 
daily for two days, for each of the five dosing courses; 10 total dosing days. One 
(100 mg) tablet will be taken in the morning, for each dosing period. Thus, 
Dasatinib will be taken orally on an intermittent, one-day schedule (starting every 
27 days) with 26-day no-therapy periods in between dosing regimens, repeated 
every 28 days over 20 weeks, resulting in ten total dosing days throughout the 
entire intervention. 
2) Quercetin is a natural-occurring flavonoid known to inhibit PI3Kinase, other kinases, and 
mTOR pathways. Quercetin is present in many fruits, vegetables, and grains and is also 
used as an ingredient in supplements, beverages, or various types of foods. Quercetin is a 
supplement and not FDA approved for any indication. The recommended dose ranges 
from a total of 750-1500 mg per day. The primary contraindication/warning is 
hypersensitivity to Quercetin. Listed drug interactions include: cyclosporine, digoxin, and 
fluoroquinolones. 
a. Dosing: Quercetin dehydrate capsules (250 mg each) equating to 1000 mg total 
daily dosage will be administered orally for three consecutive days for each of the 
five dosing periods; 15 total dosing days. Two (250 mg) capsules will be taken 
each morning and two (250 mg) capsules will be taken in the evening for each of 
the three days, within each dosing period. Thus, Quercetin will be taken orally on 
an intermittent, three consecutive day schedule (starting every 28 days) with 25-
day no-therapy periods in between dosing regimens, repeated every 28 days over 
20 weeks, resulting in five total three-day dosing regimens (15 total dosing days) 
throughout the entire intervention. 
5.3 
Method for Assigning Subjects to Treatment Groups 
Subjects who meet eligibility criteria and agree to take the assigned therapy for the entire study 
duration, will be randomized (using block randomization, as detailed below) to one of two 
groups: i) Untreated control group; ii) Dasatinib (D; 100 mg/d, for two days, Sprycel, Bristol 
Myers Squibb) plus Quercetin (Q; 1000 mg/d; 250 mg capsules x 4/d Quercetin Phytosome, 
Thorne Research). As detailed above, therapy (D+Q) will be taken orally on an intermittent 
schedule (every 28 days) with no-therapy periods in between doses, repeated every 28 days over 
20 weeks.  Block randomization (using blocks of four) will be performed by our statistician, 
 
M.S., to ensure balanced group assignment as the study proceeds. Subjects 

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will be randomized in a 1:1 ratio to the untreated control group and D+Q.  Randomization data 
will be provided to the Research Pharmacy by the statisticians. 
5.4 
Preparation and Administration of Study Drug 
The request for the study drugs (investigational products) will be sent to the Research Pharmacy 
and individually prepared for each subject. This will be managed by the Research Pharmacy 
according to their established stand procedures. Subjects randomized to the Dasatinib plus 
Quercetin (D+Q) intervention arm will be provided with one bottle containing two tablets of 
Dasatinib, and one bottle containing 12 capsules of Quercetin at outpatient CRU study visits at 
Weeks 1 (Day 1), 5 (Day 29), 9 (Day 57), 13 (Day 85), and 17 (Day 113). Study subjects will be 
instructed to take the Quercitin within 5 minutes of the Dasatinib and to ingest all the pills/capsules 
from each bottle each day. They will record their administration in the provided drug diary. 
5.5 
Subject Compliance Monitoring 
Patient adherence to study treatment will be monitored by drug accountability (pill counts). The 
study coordinator will perform the pill counts. 
5.6 
Prior and Concomitant Therapy 
Drugs listed as part of the exclusion criteria are not permitted during each of the study drug periods. 
If patients are required to initiate these medications within the study drug period, then they will be 
removed from the study primarily due to risk of drug-drug interactions. 
5.7 
Packaging 
The investigational products for this study will be delivered to, managed, and packaged by the 
Research Pharmacy according to their established standard procedures. At outpatient CRU study 
visits at Weeks 1 (Day 1), 5 (Day 29), 9 (Day 57), 13 (Day 85), and 17 (Day 113), subjects 
randomized to the Dasatinib plus Quercetin intervention arm will be provided with one bottle 
containing two tablets of Dasatinib, and one bottle containing 12 capsules of Quercetin. As noted 
above, the bottles will be prepared and dispensed by the Research Pharmacy with appropriate 
labeling to include a statement that these products are for investigational use only.  
5.8 
Masking/Blinding of Study  
In order to minimize the study bias, this study will use a process for randomized group 
assignment to either the untreated control group or to D+Q. Furthermore, the laboratories 
analyzing the collected samples and initial data will not have access as to which group (Control, 
D+Q) the samples came from. Since this is an open label study, the subject and study staff will 
be aware of which group they are in.   
5.9 
Receiving, Storage, Dispensing and Return 
5.9.1 Receipt of Drug Supplies 
The investigational products for this study will be delivered to and managed by the Research 
Pharmacy according to their established standard procedures. 
5.9.2 Storage 
Investigational products should be stored at room temperature 68° to 77°F (20° to 25°C). 

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5.9.3 Dispensing of Study Drug 
The study drug is to be used exclusively in the clinical study according to the instructions of this 
protocol and directions for use. The Investigator’s designee is responsible for providing subjects 
with the study drug and instructions for dosing and proper storage of the study drug.  
5.9.4 The Investigator’s designee will record the amount of study drug dispensed, date of 
dispensing, as well as the amount of drug returned and drug remaining. Return or 
Destruction of Study Drug 
 
 
At the completion of the study, there will be a final reconciliation of drug shipped, drug 
dispensed, drug returns, and drug remaining. This reconciliation will be logged on the drug 
reconciliation form, signed, and dated. Any discrepancies noted will be documented and 
investigated, prior to return or destruction of unused study drug. Drug destroyed on site will be 
documented in the study files. 
Table 1.  Overview of study visits. 
 
Time (Days) 
 
Visits 
Screen 
 
-90 to 1 
1 
Baseline 
(Day 1) 
 1 
2 
Week 3 
(Day15
) 
  ±5 
   3 
Week 5 
(Day 29) 
 ±5 
4 
Week 9 
(Day 57) 
±5 
5 
Week 13 
(Day 85) 
±5 
6 
Week 17 
(Day 113) 
±5 
7 
End of Week 
20 (Day 140) 
±5 
8 
   Early 
Termination 
 
Informed Consent 
X 
 
 
 
 
 
 
 
 
Verify Eligibility 
X 
X 
 
 
 
 
 
 
 
Patient History 
X 
 
 
 
 
 
 
 
 
Medication History 
X 
X 
X 
X 
X 
X 
X 
X 
X 
ECG  
X 
X  
 
X  
X  
X  
X  
X  
X  
Physical Exama 
X 
 
 
 
 
 
 
X 
X 
Screening Labsb 
X 
 
 
 
 
 
 
 
 
Vital Signsc 
X 
X 
X 
X 
X 
X 
X 
X 
X 
p16Ink4a in T Cells 
X 
 
 
 
 
 
 
 
 
Randomization 
 
X 
 
 
 
 
 
 
 
Skin AGES Reader 
 
x 
 
 
 
 
 
x 
X 
Urine tests 
 
x 
X 
X 
 
 
 
x 
x 
Buccal Swab 
 
x 
x 
X 
 
 
 
x 
x 
Bone Markersd 
 
X 
X 
X 
X 
X 
X 
X 
X 
Plasma SASP Factors 
 
X 
X 
X 
 
 
 
X 
X 
Safety Labse 
 
 
X 
X 
X 
X 
X 
X 
X 
DXA: 
full 
body, 
spine, hip, radius  
 
X 
 
 
 
 
 
X 
X>10 weeks 
HR-pQCTf 
 
X 
 
 
 
 
 
X 
X>10 weeks 
Study Drugs 
 
X 
 
X 
X 
X 
X 
 
 
Record book 
 
X 
 
X 
X 
X 
X 
X 
X 
New fractures 
 
X 
X 
X 
X 
X 
X 
X 
X 
Questionnairesg,h 
 
Xg,h 
 
Xg 
Xg 
Xg 
Xg 
Xg,h 
Xg,h 
Adverse Events 
X 
X 
X 
X 
X 
X 
X 
X 
X 
a. 
Blood pressure, temperature, respiration, height, weight, and heart rate 
b. 
Fasting blood glucose, CBC w/differential, creatinine, 25-hydroxyvitam D, AST, hemoglobin A1c, TSH, bilirubin, phosphorus, calcium, 
cystatin C, and alkaline phosphatase 
c. 
Temperature, blood pressure, pulse, weight, and respiration 
d. 
Fasting blood for plasma and cells for future research/bone markers (P1NP & CTx) 
e. 
CBC, AST, cystatin C   
f. 
High-resolution peripheral quantitative computed tomography, radius & tibia 
g. 
SF 36 QOL 
h. 
Health interview (baseline only). Geriatric depression scale and Pittsburg Fatigability questionnaires 

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6 Study Procedures 
6.1 
Visit 1 – Screening Visit 
A tabulated summary of all visits and assessments described in the following sections is provided 
in Table 1. To the extent possible, subjects will be expected to adhere to the established visit 
schedule. After discussing the study with the investigator/appropriate study staff and after 
agreeing to study participation by signing the consent, subjects will be assigned a subject 
number. For any subject, it is the responsibility of the investigator or study team member to 
obtain written informed consent (subject’s signature) prior to performing any protocol-mandated 
assessment. However, assessments performed as part of the routine care of the subject may be 
used to assess eligibility. The subject number will identify the subject throughout the study. In 
case of re-screening, the subject number assigned during the first screening procedure will be 
retained. 
 
Patients meeting the entry criteria and consenting to participate in the protocol will undergo the 
following: 
• Eligibility confirmation and informed consent discussion and documentation. 
• Review of medications. 
• Medical History. 
• EKG/ ECG – assessment of QTc interval. 
• Fasting Blood work – screening labs, peripheral blood CD3+ T cell assay for p16INK4a. 
• Vital signs (blood pressure, temperature, height, weight, respiration, and heart rate). 
• Query subject concerning adverse events. 
 
6.2 
Visit 2 – Baseline/Enrollment (Start of Week 1, Day 1) 
Patients who meet eligibility criteria during Visit 1 will attend Visit 2. This visit will be recorded 
as “Day 1” and will require a “fasting” visit to the outpatient CRTU research center at Mayo Clinic 
where the following tests will be performed: 
• Eligibility confirmation. 
• Health interview, Geriatric depression scale and Pittsburg Fatigability scale questionnaires 
• Vital signs (blood pressure, temperature, respiration, weight, and heart rate). 
• Review of medications. 
• SF 36 (QOL) Health Survey. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• Blood work (P1NP, CTx, SASP markers). 
• Skin AGES – Skin AGEs will be assessed using the AGE Reader®. 
• DXA scans (Total Body, Spine [AP], Hip, & Forearm). 
• New fracture assessment. 
• Xtreme CT high-resolution peripheral quantitative computed tomography (HR-pQCT) 
scans (distal and proximal radius & tibia). 
• ECG to monitor QTc prolongation related to dasatinib 
• Urine tests (senescent markers, urine stored for future research) 
• Buccal swab (senescent markers and biological clock) 

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• Plasma & Cells for future research. 
o This is a routine procedure and will be performed by an experienced phlebotomist 
using aseptic technique. About 100 mL of blood will be withdrawn in total, which 
poses no significant risk. Blood will be collected and stored until analysis. 
• Investigation product administration – subjects will be provided with their randomized 
study drug (D+Q) in a sealed bottle(s) from the Mayo Clinic Research Pharmacy on Day 1 
in the presence of a study coordinator. Subjects must consume all medications within the 
allotted visit time, taking Quercitin within 5 minutes of Dasatinib. Administration 
verification will be documented on the drug diary form. Subjects will be sent home with 
the remaining two days of study drugs (Day 2, 3), the drug diary form and the record book 
(for adverse event recording). Drug education will be provided by the study coordinator to 
those subjects receiving D+Q.  
• ECG to monitor QTc prolongation related to dasatinib. 
6.3 
Visit 3 – Start of Week #3 (Day15 ± 5 Days) 
This visit will be recorded as “Day 15”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of any additional medications taken since last visit. 
• New fracture assessment. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• SF 36 (QOL) Health Survey 
• Blood work (P1NP & CTx; safety labs for possible dasatinib toxicities – CBC, AST, and 
cystatin C. 
• Plasma & Cells for future research: 50 ml blood drawn to be stored until analysis 
• Urine tests (senescent markers, urine stored for future research) 
• Buccal swab (senescent markers and biological clock) 
6.4 
Visit 4 – Start of Week #5 (Day 29 ± 5 Days) 
This visit will be recorded as “Day 29”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of any additional medications taken since last visit. 
• New fracture assessment. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• SF 36 (QOL) Health Survey. 
• Blood work (P1NP & CTx; safety labs for possible dasatinib toxicities – CBC, AST, and 
cystatin C. 
• Urine tests (senescent markers, urine stored for future research) 
• Buccal swab (senescent markers and biological clock) 
• ECG to monitor QTc prolongation related to dasatinib 
• Investigation product administration – subjects will be provided with their randomized 
study drug (D+Q) in a sealed bottle(s) from the Mayo Clinic Research Pharmacy on Day 
29 in the presence of a study coordinator. Subjects must consume all medications within 
the allotted visit time, taking Quercitin within 5 minutes of Dasatinib. Administration 
verification will be documented on the drug diary form. Subjects will be sent home with 
the remaining two days of study drugs (Day 30, 31), the drug diary and the record book. 

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Drug education will be provided by the study coordinator to those subjects receiving 
D+Q. 
• Plasma & Cells for future research: 50 ml blood drawn to be stored until analysis 
6.5 
Visit 5 – Start of Week #9 (Day 57 ± 5 Days) 
This visit will be recorded as “Day 57”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of any additional medications taken since last visit. 
• New fracture assessment. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• SF 36 (QOL) Health Survey. 
• Blood work (P1NP & CTx; safety labs for possible dasatinib toxicities – CBC, AST, 
cystatin C). 
• ECG to monitor QTc prolongation related to dasatinib. 
• Investigation product administration – subjects will be provided with their randomized 
study drug (D+Q) in a sealed bottle(s) from the Mayo Clinic Research Pharmacy on Day 
57 in the presence of a study coordinator.  Subjects must consume all medications within 
the allotted visit time, taking Quercitin within 5 minutes of Dasatinib. Administration 
verification will be documented on the drug diary form.  Subjects will be sent home with 
the remaining two days of study drugs (Day 58, 59), the drug diary form and the record 
book. Drug education will be provided by the study coordinator to those subjects 
receiving D+Q. 
• Plasma & Cells for future research: 50 ml blood drawn to be stored until analysis 
 
6.6 
Visit 6 – Start of Week #13 (Day 85 ± 5 Days) 
This visit will be recorded as “Day 85”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of any additional medications taken since last visit. 
• New fracture assessment. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• SF 36 (QOL) Health Survey. 
• Blood work (P1NP & CTx; safety labs for possible dasatinib toxicities – CBC, AST, 
creatinine). 
• ECG to monitor QTc prolongation related to dasatinib. 
• Investigation product administration – subjects will be provided with their randomized 
study drug (D+Q) in a sealed bottle(s) from the Mayo Clinic Research Pharmacy on Day 
85 in the presence of a study coordinator.  Subjects must consume all medications within 
the allotted visit time, taking Quercitin within 5 minutes of Dasatinib. Administration 
verification will be documented on the drug diary form.  Subjects will be sent home with 
the remaining two days of study drugs (Day 86, 87), the drug diary form and the record 
book. Drug education will be provided by the study coordinator to those subjects 
receiving D+Q. 
• Plasma & Cells for future research: 50 ml blood drawn to be stored until analysis 
 

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6.7 
Visit 7 – Start of Week #17 (Day 113 ± 5 Days) 
 
This visit will be recorded as “Day 113”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of any additional medications taken since last visit. 
• New fracture assessment. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• SF 36 (QOL) Health Survey. 
• Blood work (P1NP & CTx; safety labs for possible dasatinib toxicities – CBC, AST, 
cystatin C). 
• ECG to monitor QTc prolongation related to dasatinib. 
• Investigation product administration – subjects will be provided with their randomized 
study drug (D+Q) in a sealed bottle(s) from the Mayo Clinic Research Pharmacy on Day 
113 in the presence of a study coordinator.  Subjects must consume all medications 
within the allotted visit time, taking Quercitin within 5 minutes of Dasatinib. 
Administration verification will be documented on the drug diary form.  Subjects will be 
sent home with the remaining two days of study drugs (Day 114, 115), the drug diary 
form and the record book. Drug education will be provided by the study coordinator to 
those subjects receiving D+Q. 
• Plasma & Cells for future research: 50 ml blood drawn to be stored until analysis 
 
6.8 
Visit 8 – End of Week #20 (Day 140 ± 5 Days) or Early Termination 
This visit will be recorded as “Day 140”. A summary of the events during this day are below: 
• Vital signs (blood pressure, respiration, temperature, weight, and heart rate). 
• Review of medications. 
• New fracture assessment. 
• Geriatric depression scale and Pittsburg Fatigability Scale questionnaires 
• 
SF 36 (QOL) Health Survey. 
• Query subject concerning adverse events: performed by a “blinded” staff member. 
• Blood work (P1NP, CTx, SASP markers; safety labs for possible dasatinib toxicities – 
CBC, AST, cystatin C). 
• ECG to monitor QTc prolongation related to dasatinib. 
• Skin AGES – Skin AGEs will be assessed using the AGE Reader®. 
• DXA scans (Total Body, Spine [AP], Hip, & Forearm). 
• Xtreme CT high-resolution peripheral quantitative computed tomography (HR-pQCT) 
scans (distal and proximal radius & tibia). 
• Plasma & Cells for future research: 100 ml blood drawn to be stored until analysis. 
• Urine tests (senescent markers, urine stored for future research) 
• Buccal swab (senescent markers and biological clock) 
 
6.9 
Assessments 
An electrocardiogram (ECG) will be conducted at screening/enrollment on all subjects and during 
visits 3-8 for the Dasatinib-randomized group members only. A timer will be set to ensure this is 

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done 2 hours after dosing. A QTc interval greater than 450 msec will require exclusion from the 
study at screening and greater than 470 msec during visits 3-8, will require withdrawal for the 
Dasatinib-randomized group members. 
6.9.1 Examinations and Procedure 
Vital signs, including pulse, blood pressure, heart rate, respiratory rate will be measured. Height 
and weight will be obtained at the pre-screening visit; weight will be obtained at each additional 
visit. Body mass index (BMI) will be recorded at screen only. 
6.9.2 Laboratory Assessments 
The following will be collected at the time points specified in the schedule of events (Section 6, 
Table 1) 
Screening Tests: 
• CBC w/diff 
• Creatinine 
• Vitamin D2 & D3 
• AST 
• Alkaline Phosphatase 
• Bilirubin 
• TSH 
• Cystatin C 
• HbA1c 
• Calcium  
• Phosphorus 
• Fasting blood glucose 
 
Blood Tests: 
• Kidney Function: creatinine, cystatin C 
• Bone Mineral Metabolism: calcium 
• Anemia: complete blood count 
• Senescence: Senescence (p16INK4a in T cells) and SASP markers (including IL-6, IL-8, 
MCP-1, Activin A) 
 
Urine Tests: 
• Senescent markers: urinary exosomes 
• Extracellular vesicles 
 
Buccal swab:  Testing will occur for determination of biological clock marker.  This is an 
epigenetic test measuring molecules and is associated with predicting epigenetic age. 
 
 
Peripheral blood CD3+ T cell assay for p16INK4a: 
10ml blood will be collected using EDTA tubes to measure p16INK4a positive lymphocyte 
population in the peripheral blood- a biomarker of senescence and chronological aging. CD3 
+lymphocyte will be collected using Whole Blood CD3, Human Microbeads (Miltenyi Biotec, 

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Cat# 130-090-874) and magnetic-activated cell sorting (MACS) machine. CD3 positive cells will 
be lysed and RNA will be isolated. P16INK4a expression will be measured by rt-qPCR using Taq-
man primer-probes, as done in Fig. 6. 
 
Future Research Studies – Plasma & Cells: 
• Collection of plasma, serum, and cells from subjects will enable us to evaluate and answer 
pertinent questions related to the biology of aging and senescence. Studies utilizing these 
samples will allow us to gain a greater understanding of the pathophysiology of accelerated 
aging and the potential for us to develop new therapies to improve patient outcomes. An 
aliquot of 100 mL plasma/serum will be obtained from each subject at Baseline and End 
of Week 20 (Day 140). The specimens will be kept on ice and immediately transported to 
Dr. Khosla’s laboratory for processing and storage. 
6.9.3 Questionnaires: 
The Short Form (36) Health Survey will be administered to subjects at each visit. This survey 
consists of 36 questions and eight scaled scores (vitality, physical functioning, bodily pain, general 
health perceptions, physical role functioning, emotional role functioning, social role functioning, 
and mental health). The Geriatric Depression Scale is a 15-item screening tool which can be used 
to identify depression in older adults.  This will be given at baseline and study end visits. If the 
GDS questionnaire is indicative of depression (score ≥ 10), the study team will make the subject 
aware of this and contact the subject’s primary care provider.  If the subject does not have a 
primary care provider, then consultation with psychiatry will be offered. If there are concerns 
regarding acute suicidal risk, the study team will contact one of the PIs who will then make 
appropriate arrangements for acute management.), diarrhea and sleep disorders.  
The health interview includes questions about general health, gynecological history, social habits, 
and fall or fracture history and will be given at the baseline visit.  The Pittsburg Fatigability Scale 
is a 10-item questionnaire that assesses self-report whole-body physical and mental tiredness 
related to activities of fixed intensity and duration in adults age ≥60.    
6.9.4 Skin AGEs Assessment: 
Skin AGEs will be assessed using the AGE Reader®.  This non-invasive method has a light source 
which illuminates a skin surface of approximately 4 cm2 on the volar side of the forearm. The 
device uses an excitation light source with peak intensity of approximately 370 nm to excite 
fluorescent moieties in the tissue (skin auto-fluorescence), which then will emit light of a different 
wavelength. In the used wavelength band, the major contribution in fluorescence comes from 
fluorescent AGEs linked mostly to collagen. Emission light and reflected excitation light from the 
skin are measured with a spectrometer in the 300–600 nm range. 
7 Statistical Plan 
In the case of an adverse event, information related to that particular patient will be revealed to the 
investigators. 
7.1 
Sample Size Determination 
Based on our recent 20-week RCT in postmenopausal women treated with β-blockers(93), we 
calculated the sample size to compare the percent change from baseline to 20 weeks in CTX  
between the senolytic (D+Q) and the untreated control group, while allowing for an estimated 

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withdrawal rate of 10% (which is realistic given the relatively short trial duration) across the two 
groups. This calculation demonstrated that 30 subjects per group will provide the study with 90% 
power to detect a 19.8% difference in the bone resorption marker, CTX (17.1% with 80% 
power), based on a two-sample t-test. 
7.2 
Statistical Methods 
 
Descriptive Statistics 
For the analyses, categorical data will be described by counts and percentiles and quantitative 
data by means and standard errors or standard deviations. Univariate descriptive statistics and 
frequency distributions will be calculated, as appropriate for all variables. Baseline values for 
demographic, clinical, and outcome variables (primary and secondary) will be tabulated for the 
different groups. Adherence to medication regimes will be described in terms of percent of 
medication taken (actual or reported) for each group. 
 
Handling of Missing Data 
All participants who are randomized will be included in the primary efficacy analysis dataset. The 
analysis will follow the intent-to-treat (ITT) principle (i.e., according to the treatment arm they are 
assigned to). For patients whose 20-week CTX and P1NP measurements are missing, data will be 
imputed. For subjects missing the 20-week measurements, individual regression models will be fit 
regressing each specific bone turnover marker (CTX or P1NP) on time (weeks); 20-week values 
will then be extrapolated based on these regression lines. 
 
 
Primary Hypothesis: In a cohort of 60 older postmenopausal women, aged ≥60years, with a 
high senescent cell burden (as these individuals are most likely to benefit from pharmacological 
interventions that eliminate senescent cells – i.e., senolytics), we hypothesize that intermittent 
senolytic therapy (D+Q) as compared to non-therapy will improve bone resorption markers over 
the course of 20 weeks. 
 
The Primary Hypothesis will be tested using an ANCOVA model with treatment as a fixed effect 
and the baseline measurement as a covariate, followed by an F-test to determine the overall 
difference between the groups. The 20-week percent changes in secondary endpoints will also be 
examined using this ANCOVA model. Although randomization should ideally obviate the need 
to correct for baseline factors, given two groups and 30 subjects per group, there is the possibility 
that randomization may not perfectly balance the subjects regarding potential covariates, such as 
the baseline values of the respective response variables. A detailed analysis of ANOVA versus 
ANCOVA by Van Breukelen(95), concluded that for RCTs with pre- and post-intervention 
measurements, both methods are appropriate and unbiased, but the ANCOVA has more power 
and is therefore recommended. All randomized subjects will be included in the primary efficacy 
analysis, which will follow the intention-to-treat (ITT) principle; i.e., according to the treatment 
arm to which they were randomized. To optimize the sensitivity of the ITT analysis, every effort 
will be made to obtain follow-up data at 20 weeks on any subject who discontinues the study at 
an earlier time point.  
 

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Secondary Hypothesis 1: In the same cohort of 60 older postmenopausal women, aged 
>60years, with a high senescent cell burden, we hypothesize that intermittent senolytic therapy 
(D+Q) as compared to non-therapy will reduce systemic surrogate markers of senescent cell 
abundance (p16INK4a in PBTLs) and circulating senescence-associated secretory phenotype 
(SASP) factors. 
 
The Secondary Hypotheses will be tested using an ANCOVA model with treatment as a fixed 
effect and the baseline measurement as a covariate. For secondary exploratory analyses, a mixed-
effects model will be used to allow the treatment effect to vary across time and to include all 
subjects (i.e., even those who did not contribute data at all time points). All subjects who receive 
any drug (D+Q) will be included in the safety and tolerability evaluations. Frequency tables will 
summarize AE occurrences by treatment arm, though group differences are likely since the 
treatment arm is known to the subjects.  
 
7.3 
Subject Population(s) for Analysis 
Analysis of the primary hypothesis will include subjects who have completed the final visit (at 
Week 20; Day 140), regardless of study drug adherence. Analysis of the secondary hypotheses 
will follow an intent-to-treat (ITT) model and will include all subjects who have completed at 
least one follow-up visit following initial randomization, again regardless of drug adherence. 
Secondary analysis will include only those subjects who completed the entire study drug course 
and were seen at all visits. 
8 Safety and Adverse Events 
Safety evaluations will include adverse event (AE) and serious AE (SAE) reporting.  Adverse 
event reporting will be conducted throughout the study for all subjects.  The reporting period 
begins at the time of informed consent and continues through study completion.  Adverse events 
will be assessed at every visit (except the screen visit) by a “blinded” staff member.   
 
Definitions, documentation and reporting of AE’s are described in Section 8.1-8.4. 
8.1 
Definitions 
Unanticipated Problems Involving Risk to Subjects or Others (UPIRTSO) 
Any unanticipated problem or adverse event that meets the following three criteria:  
• Serious: Serious problems or events that results in significant harm, (which may be 
physical, psychological, financial, social, economic, or legal) or increased risk for the 
subject or others (including individuals who are not research subjects). These include: (1) 
death; (2) life threatening adverse experience; (3) hospitalization - inpatient, new, or 
prolonged; (4) disability/incapacity - persistent or significant; (5) birth defect/anomaly; (6) 
breach of confidentiality and (7) other problems, events, or new information (i.e. 
publications, DSMB reports, interim findings, product labeling change) that in the opinion 
of the local investigator may adversely affect the rights, safety, or welfare of the subjects 
or others, or substantially compromise the research data, AND 
• Unanticipated: (i.e. unexpected) problems or events are those that are not already 
described as potential risks in the protocol, consent document, not listed in the 

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Investigator’s Brochure, or not part of an underlying disease. A problem or event is 
"unanticipated" when it was unforeseeable at the time of its occurrence. A problem or 
event is "unanticipated" when it occurs at an increased frequency or at an increased 
severity than expected, AND 
• Related: A problem or event is "related" if it is possibly related to the research procedures. 
 
 
Adverse Event 
An untoward or undesirable experience associated with the use of a medical product (i.e. drug, 
device, biologic) in a patient or research subject. 
Serious Adverse Event 
Adverse events are classified as serious or non-serious.  Serious problems/events can be well 
defined and include: 
• death 
• life threatening adverse experience 
• hospitalization 
• inpatient, new, or prolonged; disability/incapacity 
• persistent or significant disability or incapacity 
• birth defect/anomaly 
and/or per protocol may be problems/events that in the opinion of the sponsor-investigator 
may have adversely affected the rights, safety, or welfare of the subjects or others, or 
substantially compromised the research data. 
 
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 
For this study, the study treatment follow-up period is defined as 30 days following the last 
administration of study treatment.  
 
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 sponsor-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 sponsor-investigator should instruct each subject to report, to the sponsor-

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investigator, any subsequent event(s) that the subject, or the subject’s personal physician, 
believes might reasonably be related to participation in this study.  
 
Abnormal Laboratory Values 
A clinical laboratory abnormality should be documented as an adverse event if the abnormality 
changes from a value within the normal range before treatment to one outside or worse than the 
normal range after treatments. A clinical laboratory abnormality will also be documented as an 
adverse event if the abnormality meets the definition of an SAE or requires the subject to have the 
investigational product discontinued or interrupted or requires the subject to receive specific 
corrective therapy. Such changes will prompt a repeat test, telephone call to check on the subject’s 
status, a repeat visit and/or referral to the subject’s primary care physician. For example, the subject 
will receive specific corrective therapy if their Vitamin D level is <20ng/mL, as described in 
Section 4.2, Exclusion Criteria.   In the event that a subject experiences an elevated liver function 
test (AST, ALT, ALP, and/or bilirubin) greater than 2-fold over their screening/baseline 
measurement, dosing with Dasatinib should be halted and the subject will continue in the trial and 
complete the study assessments including laboratory assessments. Additional treatment and 
continued participation will be at the discretion of study investigators. 
 
 
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 an adverse event.  
 
Neither the condition, hospitalization, prolonged hospitalization, nor surgery are reported as an 
adverse event in the following circumstances: 
 
The following hospitalizations will not be considered SAE for this study: 
• A visit to the emergency department or other hospital department <24 hours, that does 
not result in admission (unless considered an important medical or life-threatening event). 
• Elective surgery planned prior to signing consent. 
• Admissions as per protocol for a planned medical/surgical procedure. 
• Routine health assessment requiring admission for baseline/trending of health status (e.g., 
routine mammogram). 
• Medical or surgical admission other than to remedy ill health and planned prior to entry 
into the study. Appropriate documentation will be obtained in these cases. 
 
8.2 
Recording of Adverse Events 
At each contact with the subject, the study team must seek information on adverse events by 
specific questioning and, as appropriate, by examination.  Information on all adverse events 
should be recorded immediately in the source document, in the appropriate adverse event section 
of the case report form (CRF) or in a separate adverse event worksheet. All clearly related signs, 
symptoms, and abnormal diagnostic, laboratory or procedure results should be recorded in the 
source document. 

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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 ultimately 
determined that the study treatment or participation is not the probable cause.  Serious adverse 
events that are still ongoing at the end of the study period must be followed up, to determine the 
final outcome.  Any serious adverse event that occurs during the Adverse Event Reporting 
Period is at least possibly related to the study treatment or study participation should be recorded 
and reported immediately. 
 
8.3 
Reporting of Serious Adverse Events and Unanticipated Problems 
When an adverse event has been identified, the study team will take appropriate action necessary 
to protect the study participant and then complete the Study Adverse Event Worksheet and log.  
The sponsor-investigator will evaluate the event and determine the necessary follow-up and 
reporting required. 
 
8.3.1 Sponsor-Investigator reporting: notifying the Mayo IRB 
The sponsor-investigator will report to the Mayo IRB any UPIRTSOs and Non-UPIRTSOs 
according to the Mayo IRB Policy and Procedures. 
 
Information collected on the adverse event worksheet (and entered in the research database): 
• Subject’s name: 
• Medical record number: 
• Disease/histology (if applicable): 
• The date the adverse event occurred: 
• Description of the adverse event: 
• Relationship of the adverse event to the research (drug, procedure, or intervention*): 
• If the adverse event was expected: 
• The severity of the adverse event: (use a table to define severity scale 1-5**) 
• If any intervention was necessary: 
• Resolution: (was the incident resolved spontaneously or after discontinuing treatment) 
• Date of Resolution: 
The Investigator will review all adverse event reports to determine if specific reports need to be 
made to the IRB and FDA. The sponsor-investigator will sign and date the adverse event report 
when it is reviewed. For this protocol, only directly related SAEs/UPIRTSOs will be reported to 
the IRB. 
 
8.3.2 Sponsor-Investigator reporting: Notifying the FDA 
The sponsor-investigator will report to the FDA all unexpected, serious suspected adverse 
reactions according to the required IND Safety Reporting timelines, formats and requirements. 
 
Unexpected fatal or life threatening suspected adverse reactions where there is evidence to 
suggest a causal relationship between the study drug and the adverse event, will be reported as a 

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serious suspected adverse reaction.  This will be reported to the FDA on FDA Form 3500A, no 
later than 7 calendar days after the sponsor-investigator’s initial receipt of the information about 
the event. 
 
Other unexpected serious suspected adverse reactions where there is evidence to suggest a causal 
relationship between the study drug and the adverse event, will be reported as a serious suspected 
adverse reaction.  This will be reported to the FDA on FDA Form 3500A no later than 15 
calendar days after the sponsor-investigator’s initial receipt of the information about the event. 
 
Any clinically important increase in the rate of serious suspected adverse reactions over those 
listed in the protocol or product insert will be reported as a serious suspected adverse reaction.  
This will be reported to the FDA on FDA Form 3500A no later than 15 calendar days after the 
sponsor-investigator’s initial receipt of the information about the event. 
 
The sponsor-investigator must also notify the FDA (and sponsors must notify all participating 
investigators) in an IND safety report of potential serious risks, from clinical trials or any other 
source, as soon as possible, but in no case later than 15 calendar days after the sponsor determines 
that the information qualifies for reporting under § 312.32(c)(1)(i)-(iv). 
 
Findings from other studies in human or animals that suggest a significant risk in humans exposed 
to the drug will be reported.  This will be reported to the FDA on FDA Form 3500A, no later than 
15 calendar days after the sponsor-investigators initial receipt of the information about the event. 
8.4 
Stopping Rules  
• All patients will be assessed during the 20 weeks of study drug administration (D+Q) and 
during each study visit following administration of each cycle of drugs. If a severe 
adverse event occurs at any time during administration of the drug regimen, a formal 
review will occur and subsequent patients will be enrolled one at a time using the same 
regimen. If three or more severe adverse events accrue, the study will be held and either a 
potential dosing regimen revisited, or discontinuation of the study protocol will occur. 
Other interventions will be as per the direction of the Mayo IRB.  If any elevations were 
to be noted (>2x the normal level), the study participant would stop treatment and have 
levels re-drawn in a month, per the clinical judgement of the investigator.  
 
8.5 
Medical Monitoring 
It is the responsibility of the Principal Investigator to oversee the safety of the study at his/her 
site. This safety monitoring will include careful assessment and appropriate reporting of adverse 
events as noted above, as well as the construction and implementation of a site data and safety-
monitoring plan (see section 10 “Study Monitoring, Auditing, and Inspecting”).  Medical 
monitoring will include a regular assessment of the number and type of serious adverse events. 
8.5.1 Internal Data and Safety Monitoring Board 
None. 

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8.5.2 Independent Data and Safety Monitoring Board 
An independent Data and Safety Monitoring Board (DSMB) will be formed to monitor the study. 
Members will be determined and a charter developed once the NIH grant has been funded. 
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 of 1996 (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 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 (long term survival status 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. Case 
reports in the form of completed checklists will be kept ensuring inclusion/exclusion criteria and 
review of adverse events/toxicity. All data requested on the CRF will be recorded; data will be 
entered directly into REDCap. 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 will be printed legibly in 
black ink. If any entry error has been made, to correct such an error, a single straight line will be 
drawn through the incorrect entry and the correct data will be entered above it. All such changes 
will be initialed and dated. Errors will not be erased and “white-out” will not be used to correct 
errors. For clarification of illegible or uncertain entries, the clarification will be printed above the 
item, initialed, and dated. If the reason for the correction is not clear or needs additional 
explanation, details will be added related to the justification for the correction. 
 
Data Management 

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The data will be housed in both hard copy case report forms (CRFs) and eCRFs through a system 
called REDCap. 
 
Data Processing 
All data will be housed and analyzed by statistician, as detailed in Section 7. 
 
Data Security and Confidentiality 
Source documents and CRFs and original consents will be stored in secured locations. All data 
will be entered into a password protected, limited access database. Individually identifiable 
patient history and medical record information will be stored in a database under coded accession 
numbers. Clinical laboratory values will be stored in the electronic medical record system, 
requiring protected password access. These data are monitored regularly for access and a formal 
policy regarding protection of personal privacy is in place. The key to identification of subjects 
will be maintained in a secure office environment under the direction of the principal 
investigators. 
 
Data Quality Assurance 
Manual and computerized quality checks will occur during data collection and analyses and any 
discrepancies will require Case Report Form (CRF) review and validation of correct data. 
 
Data Clarification Process 
To be determined. 
9.4 
Records Retention 
The sponsor-investigator will maintain records and essential documents related to the conduct of 
the study.  These will include subject case histories and regulatory documents. The sponsor-
investigator will retain the specified records and reports for: 
1. Up to 2 years after the marketing application is approved for the drug; or, if a marketing 
application is not submitted or approved for the drug, until 2 years after shipment and 
delivery of the drug for investigational use is discontinued and the FDA has been so 
notified. OR 
2. As outlined in the Mayo Clinic Research Policy Manual “Retention of and Access to 
Research Data Policy” 
   
 
Whichever is longer. 
10 Study Monitoring, Auditing, and Inspecting 
10.1 Study Monitoring Plan 
The investigator will allocate adequate time for such monitoring activities.  The Investigator will 
also ensure that the monitor or other compliance or quality assurance reviewer is given access to 
all the study-related documents and study related facilities (e.g., pharmacy, diagnostic laboratory, 
etc.), and has adequate space to conduct the monitoring visit. 

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10.2 Auditing and Inspecting 
The investigator will permit study-related monitoring, audits, and inspections by the IRB, the 
sponsor, and government regulatory agencies, of all study related documents (e.g., source 
documents, regulatory documents, data collection instruments, study data etc.).  The investigator 
will ensure the capability for inspections of applicable study-related facilities (e.g., pharmacy, 
diagnostic laboratory, etc.). 
 
Participation as an investigator in this study implies acceptance of potential inspection by 
government regulatory authorities and applicable compliance offices. 
11 Ethical Considerations 
This study is to be conducted according to United States government regulations and 
Institutional research policies and procedures. 
 
This protocol and any amendments will be submitted to a properly constituted local Institutional 
Review Board (IRB), in agreement with local legal prescriptions, for formal approval of the 
study.  The decision of the IRB concerning the conduct of the study will be made in writing to 
the sponsor-investigator before commencement of this study. 
 
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.  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 Approved IRB consent form, must be 
obtained before that subject undergoes any study procedure.  The consent form must be signed 
by the subject or the subject’s legally authorized representative, and the individual obtaining the 
informed consent. 
12 Study Finances 
12.1 Funding Source 
This study will be funded by the NIH (National Institutes of Health). 
12.2 Conflict of Interest 
Any study team member 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 
Committee-sanctioned conflict management plan that has been reviewed and approved by the 
study sponsor-investigator prior to participation in this study. 
12.3 Subject Stipends or Payments 
Remuneration will consist of up to $400.00 if subjects complete the entire study and will be 
explained in the informed consent form. Subjects will receive remuneration for each visit as 
follows: 
                                     
1. Screen  
 
 
 
 
 
$25.00 

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2. Baseline (Day 1) 
 
 
 
 
$50.00 
3. Start of Week 3 (Day 15) 
 
 
 
$25.00  
 
 
 
 
4. Start of Week 5 (Day 29) 
 
 
 
$50.00 
5. Start of Week 9 (Day 57) 
 
 
 
$50.00 
6. Start of Week 13 (Day 85) 
 
 
 
$50.00 
7. Start of Week 17 (Day 113)  
 
 
$50.00 
8. Week 20 (Day 140) or early termination 
 
$50.00 
 
Completion of the study period means that the subject followed all study-related procedures for 
each day as described in the consent form. In a token of appreciation, subjects will receive an 
additional $50.00 for completing all visits in the 20-week period. In addition, subjects will receive 
parking passes or taxi reimbursement for the time involved with completing the study visits or 
travels to the research center for study visits. Subjects will be directed where to park in order to 
receive the parking passes. 
13 Publication Plan 
The principal investigators, Drs. Khosla and Farr, hold the primary responsibility for publication 
of the results of the study. 
 
We will register with ClinicalTrials.gov prior to subject recruitment and enrollment. We will 
post results to ClinicalTrials.gov within 12 months of final data collection for the primary 
outcome. 
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