PARTICIPANT INFORMATION SHEET

You are invited to participate in a research study. This information sheet provides you with
information about the research study. The Principal Investigator (the research doctor or the person
in charge of this research) or his/her representative will also describe this research to you and
answer all of your questions. Read the information below and ask questions about anything you
don’t understand before deciding whether or not to take part.

PART I: General Information

NUS-IRB Reference Code: NUS-IRB-2021-946

1. Protocol title

Does Alpha-ketoglutarate supplementation lower BiologicaL agE in middle- aged adults

(ABLE)?

2. Principal Investigator and co-investigator(s), if any, with the contact number and
address of organization

Principal Investigator:

Andrea B. Maier, MD, PhD, FRACP

Professor, Healthy Longevity Translation Research Programme, Yong Loo Lin School of
Medicine, National University of Singapore, Co-Director, Centre for Healthy Longevity,
National University Health Systems

378 Alexandra Road, Singapore (159964)

Contact number: (65) 63703186/ 63703187 [Monday to Friday, 8.30 am to 5.30 pm]
Email: healthy_longevity@nus.edu.sg

Co-Investigators:

1. Brian Kennedy, PhD, Co-Director and Professor, NUHS Centre for Healthy Longevity,
Departments of Biochemistry and Physiology, and Healthy Longevity Translation
Research Programme, Yong Loo Lin School of Medicine, National University of
Singapore

2. Boon Seng Desmond Teo, Dr, MBBS, MRCP, MMed, GDFM, GDPM, Consultant,
Division of Advanced Internal Medicine, Department of Medicine, National
University Hospital, Consultant, Division of Advanced Internal Medicine,
Department of Medicine, National University Hospital, Consultant, Alexandra
Hospital, Assistant Professor, Department of Medicine, Yong Loo Lin School of
Medicine, National University of Singapore

3. Goh Jor Ming, PhD, Research Assistant Professor, NUHS Centre for Healthy
Longevity, Department of Physiology, and Healthy Longevity Translation Research
Programme, Yong Loo Lin School of Medicine, National University of Singapore

4. Elena Sandalova, PhD, Senior Research Fellow, NUHS Centre for Healthy Longevity,
Department of Physiology, and Healthy Longevity Translation Research Programme,
Yong Loo Lin School of Medicine, National University of Singapore

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 5. Lim Zi Xiang, PhD, Research Fellow, NUHS Centre for Healthy Longevity, Department
of Biochemistry, and Healthy Longevity Translation Research Programme, Yong Loo
Lin School of Medicine, National University of Singapore

6. Diogo Gonçalves Barardo, PhD, Research Fellow, NUHS Centre for Healthy
Longevity, and Healthy Longevity Translation Research Programme, Yong Loo Lin
School of Medicine, National University of Singapore

7. Sangeetha D/O Adiyapatham, Clinical Research Coordinator, NUHS Centre for

Healthy Longevity and Department of Physiology, Yong Loo Lin School of Medicine,
National University of Singapore

8. Lim Zhi Meng, Research Assistant, NUHS Centre for Healthy Longevity and
Department of Physiology, Yong Loo Lin School of Medicine, National University of
Singapore

9. Danica Ashrintha D/O Thanigasalam, Research Assistant, NUHS Centre for Healthy
Longevity and Department of Physiology, Yong Loo Lin School of Medicine, National
University of Singapore

10. Lim Kai Xin, Clarissa, Research Assistant, NUHS Centre for Healthy Longevity
and Department of Physiology, Yong Loo Lin School of Medicine, National
University of Singapore

11. Jessica Lu Jie Lu, PhD student, NUHS Centre for Healthy Longevity and Healthy
Longevity Translational Research Programme, Yong Loo Lin School of Medicine,
National University of Singapore

12. Rajkumar S/O Dorajoo, Senior Research Scientist, Laboratory of Metabolic Disease
and Ageing Genomics, Genome Institute of Singapore

13. Jeremy Lim, Associate Professor, Chief Executive Officer, Amili Asia

3. Whom should I call if I have any questions or problems?

Please contact the Principal Investigator, Andrea B. Maier, for all research-related
matters and in the event of research-related injuries.
Telephone: (65) 63703186/ 63703187 [Monday to Friday, 8.30 am to 5.30 pm]
Email: healthy_longevity@nus.edu.sg

This study has undergone an ethics review by the National University of Singapore Institutional
Review Board (NUS-IRB). For an independent opinion specifically regarding the rights and
welfare of research participants, you may contact a staff member of the National University of
Singapore Institutional Review Board at telephone (+65) 6516 1234 [Mondays to Thursdays
from 8.30am to 6pm, and Fridays from 8.30am to 5.30pm, except public holidays] or email at
irb@nus.edu.sg.

4. Who can participate in the research? What is the expected duration of my

participation? What is the duration of this research?

You can participate in this study if:

1) You are a middle aged (40-60 years) man or woman living in Singapore.
2) You have completed the pre-screening requirements and the research coordinator has
invited you for the screening visit.

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 3) You will be enrolled in the study (1 g AKG or placebo) if you meet the randomization
criteria after the pre-screening visit i.e., your biological age (as measured by blood
DNA methylation) is greater than your chronological age.
4) You are able to attend all 5 research visits for screening and research data collection
at the NUHS Centre for Healthy Longevity (CHL) at Alexandra Hospital: screening
visit, visit 1 (baseline), visit 2 (3 months), visit 3 (end of intervention, 6 months), and
visit 4 (post-intervention, 9 months). Each visit will last around 3 to 4 hours.
5) You have no more than ONE of the following chronic medical conditions:
a) Waist circumference more than or equal to 90 cm for males or more than or
equal to 80 cm for females
b) Fasting triglycerides more than or equal to 1.7 mmol/l
c) High-density lipoprotein less than 1.0 mmol/l in men or less than 1.3 mmol/l in
women
d) Resting blood pressure more than or equal to 130/85 mmHg or use of
antihypertensive medication
e) Fasting glucose more than or equal to 6.0 mmol/l
f) Osteopenia
g) Mild Osteoarthritis not interfering in daily activities
h) Fatty liver
6) You are willing to wear a small activity monitor for 7 consecutive days.

You will NOT be able to participate in this study if you fall in the following categories:
1) Pre-existing, or history of major CVD (coronary artery disease, heart failure, stroke,
peripheral vascular disease, pulmonary hypertension), severe/uncontrolled
hypertension (under 3 or more than 3 prescribed medications), rheumatic heart
disease, congenital heart disease, deep vein thrombosis, pulmonary embolism
2) Type 1 diabetes and Type 2 diabetes under oral metformin or insulin therapy and with
diabetic complications such as diabetic retinopathy, diabetic nephropathy
3) Active cancer or treatment of cancer in the last 3 years
4) Chronic obstructive pulmonary disease (COPD), severe asthma (taking daily
medications)
5) Pregnant women will not be recruited into this study because of the safety issues
associated with X-ray irradiation during DXA scan
6) Potential female participant who plans on pregnancy within the next 9 months of study
period
7) Multiple sclerosis and autoimmune/immune deficiency diseases such as Rheumatic
arthritis, HIV, Crohn’s disease
8) Recent history of sepsis or infection (within 3 months of in-patient hospitalization)
9) Any psychiatric disease or neurodegenerative diseases such as Alzheimer’s Disease,
Parkinson’s Disease, Lewy body dementia, and any eating disorders
10) Any metal implants in the body
11) Hepatitis and Liver cirrhosis (independent of severity)
12) Severe kidney disease (GFR less than 30 ml/min/1.73 m2)
13) Skin disease (on oral or systemic medication for immune system)
14) Subjects receiving any other similar investigational product within 60 days or 5 half-
lives before the screening, whichever that is longer
15) Any serious medical illness which in the PI’s judgment may jeopardize the subject by
his or her participation in this study or may hamper his or her ability to perform and
complete procedures required in the study

5. What is the approximate number of research subjects involved?

Our study will recruit 120 participants in total. Participants will be randomized equally
into 2 groups (n = 60 participants per group, either 1 g Ca-AKG or placebo treatment)

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6. What will be done if I take part in this research study?
If you meet the randomization criteria (your biological age is greater than the
chronological age), you will be randomized (the computer will randomly assign) into one
of the 2 groups: either 1 g (2 tablets of 500 mg) sustained-release oral Ca-AKG or placebo
daily for a total of 6 months in a double-blinded fashion (neither you nor the study team
member will know if you are given Ca-AKG tablets or placebo tablets).

Before you could be randomized into one of the two groups (daily oral 2 tablets of 500
mg Ca-AKG or 2 tablets of placebo) you need to complete the screening visit today
and meet all the eligible criteria. Today, we will be collecting your sociodemographic
data, medical history, medication records, perform some physical examinations, and
finally, 10 ml of blood will be collected to analyze biological age, blood lipids and blood
sugar. It will take around 10 to 15 days for a blood test to analyze your biological age
based on DNA methylation status. DNA methylation is a biological process that causes
chemical changes in our DNA, without changing any patterns in DNA function. Based on
such chemical changes happening in your DNA, your biological age will be estimated. If
you meet the inclusion/exclusion criteria, a study team member will contact you to
schedule for the baseline visit (visit 1) and enrol you in the study. If you do not meet the
inclusion/exclusion criteria after the blood test, the blood report test will be mailed to you.
You can then take the information back to your healthcare providers and get the
appropriate advice.

This study will be conducted at NUHS CHL, Alexandra Hospital. All biological specimens
(saliva, whole blood and stool samples) will be obtained at CHL and processed at National
University of Singapore (NUS), and all clinical/functional (e.g., lifestyle assessment,
handgrip strength and biceps/leg curl) and physiological (e.g., arterial stiffness, DEXA)
tests will be conducted at CHL.

For this study, you will be required to visit the NUHS CHL research clinic at Alexandra
Hospital 5 times: screening visit, baseline (visit 1), 3 months (visit 2), 6 months (visit 3),
and 9 months (visit 4) (see Figure 1). You will spend about 3 to 4 hours per visit at the
CHL for physiological, functional assessments and bio-specimen collection.

Figure 1. Schematic representation of study visits. Participants will be randomly assigned to receive a
placebo or 1 g Ca-AKG.

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 Study procedure at each research visit is explained in detail below:

For all 5 visits including the screening visit today, you will come to the NUHS Centre
for Healthy Longevity (CHL) at Alexandra Hospital in the morning. You would need to
fast overnight for the blood draw as fasting blood collection is required for the study. You
have to refrain from alcohol / caffeine consumption for 24 hours and abstain from dietary
supplements before all 5 visits. The study team member will inform you in advance that
overnight fasting is required. After the blood is collected, you will be provided some
crackers.

We will then check whether you are feeling fine (no fever, cold, etc.) and physically fit to
undergo the procedures listed below. If you are feeling unwell, your visit will be
rescheduled to another day. You will be requested to self-declare your health status.
a) Resting blood pressure: You will sit quietly for 5 minutes and have your blood pressure
taken by an automatic blood pressure monitor. If your blood pressure is > 160/90 mmHg,
it will be measured again after 5 minutes. If it is still > 160/90 mmHg after the second
reading, the site Principal Investigator or a physician at the Geriatric Clinic will examine
you and determine whether you should be rescheduled for another visit, or if you will be
considered ineligible for enrolment into the study.

b) Height and Weight: You will have your height and weight taken using standard
stadiometer and a digital weight scale respectively.

c) Waist and hip circumference: A staff member will measure your waist and hip
circumference with a cloth measuring tape. These measurements will be performed
twice. Female staff is available to assist ladies.

d) Dual-energy X-ray absorptiometry (DXA scan): This is a method to image your bone,
skeletal muscle and fat. It is a form of X-ray although the level of radiation is 1/100 times
of a normal chest X-ray; so, it is much lower in dosage than X-ray. The process involves
lying down on the scanning bed and this should take no longer than 30 minutes.

e) Cardiopulmonary Exercise Testing (CPET): You will be asked to exercise on a

treadmill while fitted with a face mask, a heart-rate monitor. The face mask is connected
to a metabolic cart to assess respiratory gas exchange parameters during the exercise
test. During exercise, your vital signs will be monitored at regular intervals. Finger prick
will/may be conducted up to 2 timepoints: before and after the exercise. At any point,
when the test is terminated for any reason(s), an immediate post-exercise finger prick
will be taken. The blood from the finger prick will be used to measure blood lactate and
blood glucose. Oxygen consumption, exhaled carbon dioxide, heart rate, Rate of
Perceived Exertion (RPE) will be measured. This procedure will take around within 60-
80 minutes in total. Participants may trip during the exercise test, and thus our treadmill
is equipped with a harness that will be secured to the participant that will prevent them
from falling off the treadmill belt. We will include participants based on the inclusion and
exclusion criteria. With exercise, there will always be a small risk of a cardiac event.
Hence, a potential Serious Adverse Event (SAE) would be one that triggers a cardiac
event, in which case, the study team will activate Alexandra hospital’s code blue
immediately.

f) Arterial Stiffness: You will lie comfortably on an examination bed for 15 minutes, after
which a blood pressure cuff will be placed around your arm to measure your blood
pressure. Following that, it will be removed while a blood pressure cuff will be placed
around your thigh. The research staff will then measure the distance from your neck to
your thigh with a measuring tape. This distance will inform us how fast your pulse travels

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 in your major blood vessels, which is related to how stiff the blood vessels are. The
research staff will then gently place a probe on your neck to detect the pulse. This
procedure will take 20-25 minutes. Female staff is available to assist ladies.

g) Skin autofluorescence: You will sit comfortably and place your arm on the device. The
device will shine a light into your skin to determine the levels of advanced glycation
products, which are biomarkers related to inflammation. This procedure takes less than
5 minutes.

h) Handgrip strength: You will sit comfortably in a chair and squeeze as hard as you can
on a handgrip device, with your arm bent 90 degrees at the elbow. You will be given 3
attempts to perform your best in both hands.

i) 8-repetition maximum (8-RM) Leg extension test: You will sit on a leg extension chair
with your back leaning against the backrest. You will them will perform leg extension
with the initial weights of 10 kg. If you can perform 8 repetitions, but without being able
to do the 9th repetition, the weight will be recorded as 8 RM strength. If you are able to
do more than 8 repetitions for the given weight, weights will be added every set, with an
adequate rest period, until 8 RM is achieved. The Weight increments are 2.5kg, 5kg and
100kg. For a validated repetition, a complete range of motion has to be performed.

j) Questionnaires: You will be required to complete few questionnaires to assess your

socio-demographic information, present and past medical information, quality of life,
sleep and as well as cognitive function. For most of these questions, you may answer
“Yes/No”. The questionnaires will take about 30 minutes to complete.

k) Blood: Peripheral fasting blood will be drawn from the antecubital vein by a nurse or
phlebotomist or trained research staff. Around 54.5 mL (15.5 mL for screening visit)
will be collected at each study visit. We will take your blood to see whether some
indicators of health, such as blood cholesterol, HbA1c (a measurement of the average
level of sugar in your blood over the preceding 3 months) and other markers of
inflammation are affected by the use of Ca-AKG.

l) Saliva Samples: Saliva will be collected into two polypropylene cryovial (plastic tube).
You will be instructed to let saliva collect in your mouth and gently push the saliva into a
short cryovial.

m) Stool Samples: During the research visits at CHL, the research staff will pass you the
stool containers. An instruction sheet detailing the procedure for the collection of stool
samples will also be given to you. You will return your stool sample in the stool container
within 24 hours of collection.

n) Food intake and physical activity record:

• You will be issued a commercial activity tracker to monitor your routine activity for
7 consecutive days. It can be used to estimate postures and energy expenditure
derived from different types of activities. A 3-day food diary will be emailed as a soft
copy or issued as a hard copy during each visit, and you will then record your routine
meals at least 3 times a week (2 weekdays and 1 weekend). You may email or
mail us back the food diary after completion.
• You will also be issued a physical activity diary (emailed as a soft copy or issued
as hard copy) during each visit to record the time when you go to bed, when you
wake up, or go to work, or go out of the home, as well as any physical activity start
and end time as well as intensity for 7 days. You will email or mail us the physical
activity diary after completion.

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 • the activity tracker must be returned to the study team via postage mail after
completing the procedure and all mail postage costs will be covered by study
team. If there is any damage or loss of devices, participants may have to pay 50%
of the cost of the device.

7. Will there be reimbursement for participation?

The study procedures will be performed at no charge to you. There is no cost for
participating in this study.

You will be provided SGD $30 for each visit, as transportation reimbursement.

8. How will my privacy and the confidentiality of my research records be protected?

Your participation in this study will involve the collection of “Personal Data”. “Personal
Data” means data about you which makes you identifiable (i) from such data or (ii) from
that data and other information which an organization has or likely to have access. This
includes medical conditions, medications, investigations, and treatment history.

Working data will be entered on paper forms and electronic databases (Excel, SPSS,
Qualtrics and REDCap). They will then be stored in an assigned university laptop, NUS
secure cloud (nBox, OneDrive) and data will be backed up to at least one encrypted
external hard drive. All research documents/databases will be devoid of participant
identifiers. The participant identifiers will be kept in a source file that is kept separate from
the working data. Only the PI and designated study team members will have access to the
links to identifiable source data.

To prevent unauthorized access, disclosure, or loss of data, all information collected during
the research will be secured by data encryption and passwords (for electronic data) and
under lock and key (for paper hardcopies). These data and records will only be accessible
by the PI, co-investigators, and research co-ordinators. The records will however be made
available for inspection by authorized regulatory monitors or legal authorities, as when
called for.

Information and “Personal Data” collected for this study will be kept confidential. Your
records, to the extent of the applicable laws and regulations, will not be made publicly
available. All research data collected will be kept in accordance with the National University
of Singapore’s Research Data Management Policy. Research data (without personal
identifiers) used in any publication will be kept for a minimum of 15 years before being
discarded. Where any Personal Data is collected from you, we will keep the information
confidential in accordance with the Human Biomedical Research Act and other applicable
legal rules.

However, NUS, Health Sciences Authorities (HSA), NUS Institution Review Board, and
the Ministry of Health will be granted direct access to your original data records to check
study procedures and data, without making any of your information public. By signing the
Informed Consent Form attached, you (or your legally acceptable representative, if
relevant) are authorizing (i) the collection, access to, use and storage of your “Personal
Data”, and (ii) the disclosure to authorized service providers and relevant third parties.

Data collected and entered into the Case Report Forms that are the property of NUS. In
the event of any publication regarding this study, your identity will remain confidential.

Research arising in the future, based on your “Personal Data”, will be subject to review by
the relevant institutional review board.

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 Any biological samples and/or information containing your “Personal Data” that is collected
for the purposes described in this Informed Consent Form will be stored in Singapore. Only
de-identified biological samples and/or data will be transferred out of Singapore.

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 PART II: Information on this Human Biomedical Research

9. What is the nature of this biomedical research?

This is a prospective 6-month double-blinded, placebo-controlled longitudinal
interventional study on middle-aged participants to study the effect of AKG on
biomarkers of aging, with another 3 months of post-intervention follow-up. We aim to
discover new information which may contribute to scientific and/or medical knowledge in
the geoscience field. Treatments or interventions used in this research are under
evaluation and are not currently validated as standard care or standard practice.

Definition of terms used

AKG is a compound that is formed during the Krebs cycle. The Krebs cycle is a series of
chemical reactions that occurs in every cell in our body to generate energy from
carbohydrates, fat and protein. The released AKG during energy production in the Krebs
cycle has many functions such as the increase in muscle mass, improve the bone condition
and promote faster wound healing. These functions have a beneficial effect on the
treatment of diseases such as in the heart, brain, liver, and skeletal muscle, suggesting
that AKG may lower the biological age. As we grow older, the level of AKG in our body
also decreases; hence, we may need to take AKG in other forms as a dietary supplement.

Chronological age is the amount of time that has passed from your birth to the given
date. It’s your age in terms of years, months, days, etc. This is the primary way people
define their age. It’s also a primary risk factor for chronic diseases, mortality, and any
impairments to bodily functions, such as hearing and memory.

In contrast, biological age refers to how old a person seems. Also known as physiological
or functional age, biological age differs from chronological age because it takes into
consideration a number of factors other than just the day you were born. The basic idea
behind biological aging is that aging occurs as you gradually accumulate damage to
various cells and tissues in the body.
In this study, we intend to measure other health indicators such as your body strength, the
health of your arteries and blood tests that are related to your biological age, which can be
modulated using Ca-AKG. Overall, we aim to study the anti-aging function of AKG.

10. What is the purpose of this biomedical research?

The purpose of this study is to is to evaluate the anti-aging function of AKG and
investigate whether AKG can reduce biological age in middle-aged adults in Singapore.
We aim to study if AKG will affect DNA methylation (chemical changes in DNA which is
related in aging) which will be associated with the change in blood tests and body function.

This is a 6-month double-blinded, placebo-controlled longitudinal interventional study on

middle-aged participants to study the effect of AKG on biomarkers of aging, with another
3-month of post-intervention follow-up. The total duration of participation in this study
is 9 months. The rationale for this study design is to study the long-term effect of 1 g Ca-
AKG in middle-aged adults.

The primary purpose is to study the effect of 1 g Ca-AKG treatment, compared with
placebo, on DNA methylation status in middle-aged adults in Singapore.

The secondary purpose is to study the effect of AKG on arterial stiffness, body
strength and composition as well as changes in blood concentration of systemic
inflammatory markers and metabolites associated with aging.

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11. What are the possible risks, discomforts, or inconveniences to me if I participate in
this research?

There is no known side effect associated with this study procedure. While this study is
considered and is rigorously planned to minimize all potential adverse events, no study is
completely risk-free. The anticipated risks to the participants are as follows:
a) Ca-AKG: AKG is apparently safe, when taken up to 6 g per day for 6 months. While
no known adverse effects of AKG supplementation have ever been reported,
participants may experience rare side effects like nausea and abdominal discomfort,
which will be documented and reported to the Principal Investigator. The Principal
Investigator will decide whether to withdraw the participant based on his/her clinical
judgment.
b) Blood draw: There is some discomfort during venepuncture. There is also a risk of
bruising, but this should resolve on its own over a few days.
c) DXA scan: There is minimal exposure to X-rays during this scan. However, the
exposure is 100-times lower than that of a regular chest X-ray.
d) AGE reader: There is minimal exposure to UV-ray on your arm. However, the
exposure is for about 12 seconds, which is less than the amount that you would expect
from standing in outdoor sunlight for a few minutes.
e) Emotional distress: You may experience some emotional distress during this
interview. Trained counsellors will be present at all times to provide counselling and
support. You can refuse to answer any questions that you are uncomfortable with and
choose to withdraw from this study at any point by informing the interviewers.
f) CPET: Participants may trip during the exercise test, and thus our treadmill is equipped
with a harness that will be secured to the participant that will prevent them from falling off
the treadmill belt. We will include participants based on the inclusion and exclusion criteria.
With exercise, there will always be a small risk of a cardiac event. Hence, a potential
Serious Adverse Event (SAE) would be one that triggers a cardiac event, in which case,
the study team will activate Alexandra hospital’s code blue immediately. If the SAE is either
life-threatening or results in death, the study team will inform RCIO and MOH within 7 days
of the event.
g) 8-RM leg extension test: Participants may feel some soreness the day after the leg
extension test, particularly in the front of their thighs. This is normal as they are not used
to the movement, but will unlikely remain sore for more than 2-3 days. If a participant
continues to feel extreme soreness a day after the test or if the pain continues for more
than 3 days, she can contact the study team, where we will document this as anote-to-file,
and update IRB accordingly. The PI will reach out and discuss with the participant about
the case and will refer for further medical follow-up if necessary. Like in the case of CPET,
in the unlikely event that the 8-RM test triggers a cardiac event, the code blue protocol will
be followed. If the SAE is either life-threatening or results in death, the study team will
inform RCIO and MOH within 7 days of the event.
In case of any unforeseen circumstances, the research team will also ensure that the PI
(Andrea B Maier) and clinically trained Co-investigators are within the vicinity of the testing
site on the study visit days.

12. What benefits can I expect from participating in the research?

There is no known benefit from participation in this study. The bulk of the investigations
performed in this study is for research purposes and not for clinical management. Although
there may be no direct personal benefit to the participant arising from this study and the
examinations, the results of this study may be beneficial to understand the process of
aging and discover the potential treatment. This may help chart the future for healthy aging
and longevity.

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 Potential benefits include improved aging-related cardiovascular function (arterial
stiffness), bone mass density and skeletal muscle mass, after AKG supplementation.
Research findings from this study and tests results such as bone mineral density, body
mass index, blood sugar, blood lipids, total blood count, pulse wave velocity will be
provided to you. These findings can be further consulted with your treating physicians. The
full study results will be provided only after the study has been completed, unless in the
event of incidental findings.
13. Are there any alternative procedures or treatments available to me? What are the
potential benefits and risks of such alternatives?

There are no alternative procedures or treatment available to you in this study. If you
choose not to take part in this study, you will still receive standard care for your condition,
if applicable.

14. If I am injured as a result of participating in this research, what are the compensation
and treatments available to me?

It is unlikely that you will be injured during the course of this study. If you follow the
directions of the PI/research staff in charge of this study and you are injured due to the
tests conducted, NUS will pay the medical expenses for the treatment of that injury without
any legal commitment on your part to prove NUS is at fault. However, there are conditions
and limitations to the extent of compensation provided. You may wish to discuss this with
your Principal Investigator. By signing this consent form, you will not waive any of your
legal rights or release the parties involved in this study from liability for negligence.

15. Do I have to incur any expenses by participating in this research?

The study procedures will be performed at no charge to you. There are no costs for
participating in this study. The trial material (1g Calcium-Alpha-ketoglutarate or placebo
tablets) will be provided to you at free of cost. Your transportation costs will be reimbursed
as an honorarium

16. What will happen to the biological material taken from me?

You consent to having your biological samples (saliva, blood and stool) taken, which will
be analyzed to look for biomarkers that may be associated with aging. Biomarkers are
distinctive biological characteristics; whose presence indicates either normal or diseased
processes in the body. Biomarkers can be specific cells, molecules, enzymes, hormones,
genes, and physical characteristics, etc. Biomarker analysis involves analyzing genetic
material and testing of molecules. Leftover of the samples will go under the charge of the
Principal Investigator for long-term storage. Samples will be stored until they are fully
utilized or reach their expiry for storage for use in future research related to aging and age-
related diseases. Your samples will be kept for a minimum of 15 years, to facilitate future
research related to aging and age-related diseases. Additional tests may be conducted in
case we discover that there is the substance we do not currently know about that might
help us better understand or develop treatments that delay aging that is not known at the
present time. They will not be used for restricted human biomedical research/future
restricted human biomedical research involving human-animal combinations.

The biological samples collected for the study will be deemed to be gifted to the study
team and will not be returned to you. You will also not have any right or claim to any share
in the commercial gain derived from the research (if any). If you have consented to
contribute your leftover biological samples and research data for future research, your
biological samples will be de-identified and be assigned a unique subject identifier. The

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 individual performing the tests will not be informed regarding your identity. Your de-
identified information and personal identifiers will be placed in separate and secured
databases. However, you retain your right to ask the Principal Investigator to discard or
destroy any remaining samples if the biological sample(s) is noted-identified and has not
been used for the research/future research or it has been used for research, but it is
practicable to discontinue further use of the biological sample(s) for the research/future
research. We treasure the samples donated by you and we will maximize the usage till
they are fully utilized or reaches their expiry for storage. Collected biological samples will
only be used for current and future research related to aging and age-related diseases not
for any other purposes.

Your name will be removed from all data collected. De-identified data will be stored long
term under the care of the Principal Investigator after central reading and will only be
accessible to the study team or potential collaborators. Only de-identified information will
be used for analysis.

Any biological samples and/or information containing your “Personal Data” that is collected
for the purposes described in this Informed Consent Form will be stored in Singapore. Only
de-identified biological samples and/or data will be transferred out of Singapore, if
required. Biological samples transferred out of Singapore may contain genetic information
for genetic sequencing. To minimize any risk of re-identification through genetic
information, research team will secure the dataflow and restrict analysis to their accredited
laboratories only. De-identified samples will be used for future research conducted by the
healthy longevity programme and its collaborators (only). That might include genetic
testing, such as Genome Wide Association Studies. Only de-identified samples will be
used for any future research, including genetic testing.

17. Will my participation in this research involve the use of any information that will
identify me?
1. Your personal data (i.e., name, date of birth and partial NRIC, contact details) will be
collected for verifying your identity and contacting you for the purpose of the study.

18. How will my personal identifiers collected from me be kept confidential?

To prevent unauthorized access, disclosure, or loss of data, all information collected in the
course of the research will be secured by data encryption and passwords (for electronic
data) and under lock and key (for paper hardcopies). Only the principal investigator, co-
investigators and authorized research co-ordinators will have access to your personal
data, and this will not be released to any other person, including members of the research
team. Your personal data will never be used in a publication or presentation. To protect
your confidentiality, your research data will be coded (i.e., only identified with a code
number). All personal data (e.g., names and contact information) will be kept separate
from the research data. All participating subjects will be requested to provide the contact
details of a person who can be contacted if needed, and this would be kept separate from
the research data also. The link between your personal data and the code number will be
kept confidential by the principal investigator or a trusted third party.

Your identifiable information (e.g., names, Partial IC nos.) will not be disclosed, unless
required by the applicable laws and regulations (e.g., Infectious Disease Act) or for
inspection by authorized regulatory monitors or legal authorities, as when called for.

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19. Will any identifiable information obtained from me be used for future biomedical
research?

If you consent, your personal data (i.e., name, date of birth and partial NRIC) will be
retained for a period of 25 years so that we may contact you for participation in future
research and/or follow-up in this study.

20. Will I be re-identified in the event of incidental finding(s) arising during the
biomedical research?

During the course of this study or future studies, there is a possibility that we might
unintentionally come to know of new information about your health condition from the blood
tests or other clinical tests that are that are unrelated to our research. These are called
“incidental findings”. “Incidental findings” are findings that have potential health or
reproductive importance to participants like you and are discovered in the course of
conducting the study, but are unrelated to the purposes, objectives or variables of the
study. These findings may affect your future life and/or health insurance coverage.

You will be asked to indicate whether you wish to be re-identified and notified in the case
of any important incidental finding that is related to you. The discovery of an incidental
finding, at the discretion of the University, will be communicated to you for the purpose of
seeking medical advice or treatment. In the event that you have indicated not to be re-
identified and notified, but the University has determined that the incidental finding is of
clinical significance, you may still be contacted to decide if you wish to be notified of the
incidental finding at that time

If you agree to be re-identified and notified, your study doctor/a qualified healthcare
professional will explain the incidental finding to you and discuss and advise you on the
next steps to follow. You may wish to do more tests and seek advice to confirm this
incidental finding. The costs for any care that will be needed to diagnose or treat an
incidental finding would not be paid for by this research study. These costs would be your
responsibility.

21. Under what circumstances will I be re-contacted for further consent?

You will be re-contacted for further consent, including but not limited to changes in the
proposed research, serious adverse events that would lead to a change in the proposed
research.

22. Can I withdraw my consent to the research at any time?

Your participation in this study is completely voluntary. You may stop participating in this
study at any time without giving any reason. Your decision not to take part in this study or
to stop your participation will not affect your medical care or any benefits to which you are
entitled. If you decide to stop taking part in this study, you should inform the Principal
Investigator.

However, the data that has been collected until the time of your withdrawal will be kept
and analyzed. The reason is to enable a complete and comprehensive evaluation of the
study. The reason for the withdrawal will be requested, however you are not bound to
provide a reason for withdrawal. There will be no penalties or damages imposed on you
should you withdraw your consent to participate in this research. Once you are withdrawn
from this study, no attempt will be made to further evaluate you or to collect additional data
from you.

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 Your doctor and the Investigator of this study may stop your participation in the study at
any time if they decide that it is in your best interests. They may also do this if you do not
follow the instructions required to complete the study adequately. If you have other medical
problems or side effects, the doctor and/or nurse will decide if you may continue in the
research study.

In the event of any new information becoming available that may be relevant to your
willingness to continue in this study, you (or your legally acceptable representative, if
relevant) will be informed in a timely manner by the Principal Investigator or his/her
representative.

PART III: Information on the Removal and Use of My Tissue(s) for this
Research

1. What is the tissue(s) that will be taken from me and what will they be used for?

a. Blood Biomarkers
Peripheral blood will be drawn from the antecubital vein by a nurse/ phlebotomist/ trained
SAF medic, with the participant having fasted overnight, and a maximum of 54.5 mL of
blood will be collected into blood collection tubes. This would be used for Immune
parameters, metabolic, hematological, liver and renal function.

Justification for the required volumes for blood samples

Test(s) Volume and type of tube(s)

Peripheral Blood Mononuclear Cells (PBMC) 3 x 9 mL anticoagulant tubes = 27 mL
Full Blood Count (FBC) and HbA1c 1 x 3 mL anticoagulant tubes = 3 mL
Glucose Fasting 1 x 2 mL Sodium fluoride tube = 2 mL
Insulin, lipid, and protein panel 1 x 5 mL Plain tube (Gold top) = 5 mL
Plasma and Buffy Coat Cells 1 x 8.5 mL anticoagulant tubes = 8.5 mL
Serum 1 x 9 mL Plain tube = 9 mL
Total 54.5 mL (8 tubes)
Table 2. Breakdown of the blood tubes and their intended uses.

b. Saliva Samples
Saliva will be collected into two polypropylene cryovials. The subjects will be instructed to
let saliva collect in their mouth and gently push the saliva into a sterile, disposable funnel
fitted to the cryovial. The samples will be stored at – 80°C. This would be used for DNA
methylation.

c. Stool Samples
During the research visits at CHL, the research staff will pass the stool collection kit to the
participants. An instruction sheet detailing the procedure for the collection of stool samples
will be included in the kit. The participants will then return the stool container with the
samples collected within 24 h. This would be use for microbiome analysis.

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2. Will my tissue(s) be used for any purpose other than current research?

Archival de-identified samples will be used for future research related to aging and age-
related diseases research will be conducted by the healthy
longevity programme and its collaborators, conducted by the healthy longevity
programme and its collaborators (only). That might include genetic testing, such as
Genome Wide Association Studies. Only de-identified samples will be used for any future
research, including genetic testing.

3. What are the proposed area(s) of research approved by the Institutional Review
Board (IRB), where the IRB has waived the requirement that the removal of my
tissue(s) be for therapeutic or diagnostic purpose?

De-identified Samples will not be used for therapeutic or diagnostic purposes, they will
only be used for the purpose of this study.

4. What risks, discomforts and inconveniences will I expect if my tissue(s) is/ are
removed from me?

You may experience some pain and bruising when blood is collected from your vein. Fainting
is an occasional adverse event due to blood-taking. Appropriate first aid will be administered
should any of these events occur.

5. Will I still retain my rights to my tissue(s) after it has/ have been removed from me?

Donation of your tissue(s) is voluntary. By donating your tissue(s), you will relinquish your
rights to your tissue(s) and any intellectual property rights that may be derived from the use of
your tissue(s).

6. If I am injured as a result of donating my tissue(s), what are the compensation and

treatments available to me?

(For research studies conducted in Singapore only) If you follow the directions of the PI in
charge of this research study and you are injured during the donation of your tissue(s), the
NUS will pay the medical expenses for the treatment of that injury. By signing this consent
form, you will not waive any of your legal rights or release the parties involved in this study
from liability for negligence.

Injuries sustained due to this research will be covered by the NUS Clinical Trial Insurance
Policy.

7. Do I have to incur any expenses if I donate my tissue(s) for this research?

No. you do not have to incur any expenses for donating tissues for this research.

8. Will my donated tissue(s) be used in any ways that can identify me?

Biological samples collected from the participants will be coded with the same unique
individual identifiers assigned to each participant. These de-identified biological samples
will be stored within the NUS and can only be accessed by qualified laboratory personnel
under the supervision of the principal investigator or members of the study team.
Biological samples transferred out of Singapore may contain genetic information for
genetic sequencing. To minimize any risk of re-identification through genetic information,

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 research team will secure the dataflow and restrict analysis to their accredited laboratories
only. Samples will be stored until they are fully utilized or expired. Only de-identified
samples/data will be sent to external/overseas collaborators.

There may be potential use of the de-identified data and samples for future research, but
this will only apply to samples/data of participants who have consented to allow such
future usage of their samples/data.

9. Will the records that identify me in relation to my tissue(s) donation be kept

confidential?

The PI will be responsible for the confidentiality of all study information. PI will ensure that
the participant’s identity is not made publicly available. To maintain subjects’
confidentiality, all case report forms/data collection forms will be devoid of personal
identifiers. Study participants will be identified only by unique identification numbers. All
physical records will be kept locked. The research database will be password-protected
and only accessible to authorized members of the study team. Linked personal data (e.g.,
name, date of birth, email address and telephone number) will be kept in a separate file
under lock and key.

10. Will the identifiable information obtained from me in the course of tissue donation
be used for future research?

Archival de-identified samples will be used for future research conducted by the PI and
other team members in the healthy longevity program or other external collaborators.

11. What will happen to the tissue(s) taken from me upon completion of the research?

The tissues will be stored for a minimum of 15 years, for current and future research.

12. Will my tissue(s) be used in restricted human biomedical research?

Your tissue(s) will not be used in restricted human biomedical research involving human-
animal combinations.

13. Will my tissue(s) be exported overseas or removed from Singapore?

De-identified samples will be used for future research conducted by the healthy longevity
programme and its collaborators (only). That might include genetic testing, such as
Genome Wide Association Studies. Only de-identified samples will be used for any future
research, including genetic testing. De-identified samples/data will be sent to
external/overseas collaborators for the purpose of analysis for this study.

14. Will I be re-identified in the case of incidental finding(s) arising from the use of my
tissue(s) in future research?

It is possible that during the research we may find you have pre-existing medical issues
that are unrelated to our research. These are called “incidental findings”. “Incidental
findings” are findings that have potential health or reproductive importance to participants

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 like you and are discovered in the course of conducting the study, but are unrelated to the
purposes, objectives or variables of the study.

You will be asked to indicate whether you wish to be re-identified and notified in the event
of an important incidental finding that is related to you. The discovery of an incidental
finding, at the discretion of the University, will be communicated to you for the purpose of
seeking medical advice or treatment. In the event that you have indicated not to be re-
identified and notified, but the University has determined that the incidental finding is of
clinical significance, you may still be contacted to decide if you wish to be notified of the
incidental finding at that time.

After the end of study (visit 4; when participants have completed 3 months of post-
intervention follow-up), each participant will be given a choice of whether to be informed
if his or her biomedical test results in the study fall outside the range of normal values, in
which case, the results will be considered as incidental findings. If he or she has opted
and consented to be re-identified and notified, the PI will contact the participant directly to
discuss the necessary follow-up procedures with their primary care physician. There will
be no further follow-up if no incidental findings are noted.

15. Will I be re-contacted for further consent for the use of my tissue(s)? If so, under
what circumstances?

No, you would not be contacted for further consent for the use of tissue as consent has
been given initially.

16. Can I withdraw my consent to the removal of my tissue(s) and/or use of my tissue(s)
in research at any time?

You can also withdraw from the research at any time without giving any reasons, by informing
the principal investigator and all your tissue(s) collected will be discarded. However, this only
applies if the tissue is individually-identifiable and has not been used for research. Please also
note that the withdrawal of consent does not affect the research information obtained before
the consent is withdrawn and such information may still be retained and used for research.
There will be no penalties or damages imposed on you should you withdraw your consent to
participate in this research.

You are entitled to refuse to participate or discontinue participation at any time in this research.
Refusal to participate or withdrawal from participation will not affect your medical management
or cause loss of benefits to which you are otherwise entitled.

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 Consent Form for Research Subjects

Protocol title:
Alpha-ketoglutarate supplementation lowers BiologicaL agE in middle-aged adults (ABLE)

Principal Investigator with the contact number and organization:

Andrea B. Maier, MD, PhD, FRACP
Professor, Healthy Longevity Translation Research Programme, Yong Loo Lin School of
Medicine, National University of Singapore
378 Alexandra Road, Singapore (159964)
Contact number: (65) 87142729 / 87142730
Email: a.maier@nus.edu.sg

I hereby acknowledge that:

1. My signature is my acknowledgement that I have agreed to take part in the above

research.

2. I have received a copy of this information sheet that explains the use of human biological
material / data in this research. I understand its contents and agree to donate my biological
material / data the use of this research.

3. I can withdraw from the research at any point of time by informing the Principal Investigator
although if I have donated tissue for the research, I can only withdraw my consent to use
my tissue if the tissue is individually identifiable and has not been used for research or it
is practicable to discontinue further use of my tissue for research. I am aware that that the
withdrawal of consent does not affect the research information obtained before the consent
is withdrawn and such information may still be retained and used for research.

4. I will not have any financial benefits that result from the commercial development of this
research.

5. For health information and/or human biological materials collected for use in future
research related to aging and age-related diseases research will be conducted by the
healthy longevity programme and its collaborators (which might include genetic testing),
please select 1 of the 4 choices below:
I agree to donate my coded human biological material / data collected for this research
to be used in future research. The future research will be subject to an Institutional
Review Board’s approval. OR
I agree to donate my human biological material / data collected for this research to be
used in future research. However, I would like all personal data removed from my
biological samples / data so that no one can link the samples / data to me. The future
research will be subject to an Institutional Review Board’s approval. OR
I agree to donate my human biological material / data collected for this research to be
used in future research which may include testing and development by commercial
firms. However, I would like all personal data removed from my biological samples /
data so that no one can link the samples / data to me. OR
I do not agree to donate my human biological material / data collected for this research
to be used in future research. In addition, I would like to have left over biological
samples destroyed after the completion of this research.

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6. For purposes of re-contact for future research, please indicate:
I agree to be re-contacted for future related studies. I understand that future studies
will be subject to an Institutional Review Board’s approval.
I do not agree to be re-contacted for future related studies. I understand that future
studies will be subject to an Institutional Review Board’s approval.

7. For the purpose of re-contact for further consent

I agree to be re-contacted for further consent under the circumstances identified in the
Participant Information Sheet.

I do not agree to be re-contacted for further consent under the circumstances identified
in the Participant Information Sheet.

8. For purposes of re-contact and notified in the case of an incidental finding, please indicate

I agree that I may be re-contacted and notified in the case of an incidental finding from
this research or any other medical issue is found during the course of this study or in future
research, in order to facilitate my seeking proper medical advice. I understand that my
participation in the research is not considered medical treatment, and that the researchers
are not qualified medical practitioners able to provide diagnoses

I do not agree I may be re-contacted and notified in the case of an incidental finding
from this research or any other medical issue is found during the course of this study or in
future research, in order to facilitate my seeking proper medical advice. I understand that
my participation in the research is not considered medical treatment, and that the
researchers are not qualified medical practitioners able to provide diagnoses

9.

This research has been explained to me in ______________________ <State language>,

which I understand, by ___________________________________ <Name of translator>

on _________________________ <Date: dd-mm-yyyy>.

Name (Research Participant) Signature (Research Participant) Date <dd-mm-yyyy>

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I, the undersigned, certify to the following:

(a) I am 21 years of age or older.

(b) I have taken reasonable steps to ascertain the identity of the research subject.
(c) To the best of my knowledge, the research subject had the study fully explained in a
language understood by him/ her and clearly understands the nature, risks and
benefits of his/ her participation in the study.
(d) I have taken steps to ascertain that the consent has been given voluntarily without any
coercion or intimidation.

Name (Witness) Signature (Witness) Date <dd-mm-yyyy>

Note that the requirement of a witness is waived if the research is not invasive, not
interventional and is not restricted HBR. (e.g., research that comprises solely of a survey or
collection of information from research subjects)

Name (Consent Taker) Signature (Consent Taker) Date <dd-mm-yyyy>

The study has been explained to the research subject in ______________ [State Language]
☐ Check if this section is not applicable

Name (Translator) Signature (Translator) Date <dd-mm-yyyy>

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