A Proof-of-concept Clinical Study to Evaluate the Impact of a Combination Nutraceutical Kit on Muscle Function, Cognitive Performance, and Immune Resilience in Older Adults.
Phase: PHASE4
Conditions: Age-related Functional Decline, Mild Cognitive Impairment
Sponsor: GPLIFE HEALTHCARE PRIVATE LIMITED

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 1 of 46 
STUDY PROTOCOL 
A proof-of-concept clinical study to evaluate the impact of a combination nutraceutical on 
muscle function, cognitive performance, and immune resilience in older adults. 
 
 
Study Code: GP/CT/25-26/001 
Protocol Version No.: 1.00 
 
Date: 19/11/2025 
 
 
Supersedes Version No.: N/A 
Date: N/A 
 
 
 
 
 
 
 
 
 
CONFIDENTIALITY STATEMENT 
 
The information contained in this document is the property of the study sponsor. No part of it 
may be transmitted, reproduced, published, or used by another individual or organization 
without prior written authorization from the study sponsor. This document is therefore 
provided to you in confidence as an investigator, potential investigator or consultant, for 
review by you, your staff, and an applicable IRB. It is understood that this information will 
not be disclosed to others without written authorization from the sponsor, except to the extent 
necessary to obtain an informed consent document from those participants to whom the 
investigational product may be administered. 
 
 
SPONSOR DETAILS 
 
GPLIFE HEALTHCARE PRIVATE LIMITED 
Office no. 705-706, 7th floor, Orbit 1 Building, near RRTM Market, 
Punagam Saroli Road, Surat, Gujarat 395010 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 2 of 46 
 
1. STUDY APPROVAL AND MANAGEMENT TEAM 
1.1. SPONSOR’S APPROVAL 
I, the undersigned, have read and understood this protocol, and hereby agree to abide by the 
pertinent responsibilities of the sponsor for the conduct of the study in accordance with the 
WMA Declaration of Helsinki, current Protocol, ICH-GCP, ICH guidelines for Good Clinical 
Practice and local regulatory guidelines.               
 
No changes in the protocol will be implemented without formal authorization by the ethics 
committee (IRB) and regulatory authority unless there are logistic or administrative changes.  
 
I further assure that the investigational products to be used in this study are manufactured as 
per the current Good Manufacturing Practices. 
 
 
 
 
 
 
 
 
____________________________                                                         _________ 
 
             
Sponsor’s Representative                                                                      Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 3 of 46 
2. 
TABLE OF CONTENT 
SN 
Chapter details 
Page No. 
1 
STUDY APPROVAL AND MANAGEMENT TEAM 
 
2 
TABLE OF CONTENT 
 
3 
LIST OF TABLES 
 
4 
LIST OF ABBREVIATIONS 
 
5 
STUDY SYNOPSIS 
 
6 
INTRODUCTION 
 
7 
STUDY RATIONALE  
 
8 
STUDY OBJECTIVES AND ENDPOINTS 
 
9 
INVESTIGATIONAL PLAN  
 
10 
SAFETY CONSIDERATIONS 
 
11 
STATISTICAL CONSIDERATIONS AND EFFICACY 
EVALUATION 
 
12 
DOCUMENTATION 
 
13 
ETHICAL CONSIDERATIONS 
 
14 
RECORDING OF DATA, REPORT PREPARATION, AND 
ARCHIVAL 
 
15 
TERMINATION OF THE STUDY 
 
16 
PUBLICATION 
 
17 
IPR VALUE 
 
18 
REFERENCES 
 
19 
APPENDICES 
 
 
3. 
LIST OF TABLES 
Table No. 
Title 
Page No. 
1 
List of Abbreviations 
3 
2 
Study Synopsis 
5 
3 
Active Ingredients of Investigational product  
12 
4 
Efficacy Endpoints 
18 
5 
AE evaluation based on severity, causality, and outcome 
31 
6 
Schedule of study events 
49 
 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 4 of 46 
4. 
LIST OF ABBREVIATIONS 
Table 1: List of Abbreviations 
Abbreviation 
Full Form 
ICH  
International Council For Harmonisation Of Technical Requirements 
For Pharmaceuticals For Human Use 
GCP  
Good Clinical Practice 
WMA 
World Medical Association 
IRB 
Institutional Review Board 
AE 
Adverse Event  
SAE 
Serious Adverse Event 
SPSS 
Statistical Package For The Social Sciences 
PP 
Per Protocol 
IP 
Investigational Product 
CRF 
Case Report Form 
ICF 
Informed Consent Form 
ADL 
Activities Of Daily Living 
MedDRA 
Medical Dictionary For Regulatory Activities 
LAR 
Legally Acceptable Representative 
PIS 
Participant Information Sheet 
QA 
Quality Assurance 
IPIB 
Investigational Product Information Brochure 
SAP 
Statistical Analysis Plan 
SOP 
Standard Operating Procedure 
SPIRIT 
Standard Protocol Items: Recommendations For Interventional Trials 
IPR 
Intellectual Property Rights 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 5 of 46 
5. STUDY SYNOPSIS 
 Table 2: Study Synopsis 
Study Title 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical kit on muscle function, cognitive performance, and immune 
resilience in older adults. 
Study Design 
A single-arm, open-label, exploratory pilot clinical study. 
Clinical Phase 
Proof of concept 
Treatment Arm 
Single arm (Interventional nutraceutical formulation) 
Dosage 
30 g sachet content to be mixed with water and consumed twice daily 
before meals. 
Objectives 
The objectives of the study will be to: 
1. To evaluate the impact of the combination nutraceutical kit on muscle 
function, including strength, endurance, and muscle mass. 
2. To assess effects on cognitive performance, including executive 
function, memory, and processing speed. 
3. To evaluate immune resilience and inflammation modulation 
through relevant immune biomarkers and immune cell measurements. 
4. To assess safety, tolerability, and compliance of the combination 
nutraceutical kit. 
Efficacy 
Endpoints 
Assessment of changes in: 
Domain 
Test and 
Method 
Outcome Measure 
Time frame 
Primary 
Endpoint 1 
 
6-minute walk 
test 
6-minute walk 
distance (6MWD) 
(meters) 
Screening, 
and Day 60. 
Secondary 
Endpoint 1 
 
1-RM strength 
test (knee 
extension) 
1 RM (kg) by leg 
press 
Screening, 
and Day 60. 
Secondary 
Endpoint 2 
 
DXA scan  
Muscle volume 
Screening, 
and Day 60. 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 6 of 46 
 
Secondary 
Endpoint 3 
 
NIH Toolbox 
Fluid Composite 
Fluid Composite score 
Screening, 
and Day 60. 
executive function, 
attention and 
processing speed, 
working memory 
Secondary 
Endpoint 4 
 
Cognitive 
Biomarkers 
p-tau181 
Screening, 
and Day 60. 
Secondary 
Endpoint 5 
 
Immunological 
markers  
CD4/CD8 ratio, 
CD45, CD3, CD4, 
CD8, T cells, B cells, 
NK cells, 
lymphocyte/neutrophil 
ratio 
Screening, 
and Day 60. 
Secondary 
Endpoint 6 
 
Inflammatory 
markers 
hs-CRP, TNF-alpha, 
IL-6 
Secondary 
Endpoint 7 
 
Longevity and 
Putative Energy 
Markers  
MDA, SOD, NAD+ 
Screening, 
and Day 60. 
Secondary 
Endpoint 8 
 
Anthropometric 
parameters 
Body Weight (kg), 
BMI (Kg/m2) and hip, 
waist, chest 
circumferences (cm) 
Screening, 
and Day 60. 
Secondary 
Endpoint 9 
 
Hematological 
& biochemical 
investigations  
Complete blood count 
(CBC), Renal 
Function Test (RFT). 
Liver Function Test 
(LFT), Thyroid 
profile, lipid profile, 
electrolytes. 
Screening, 
and Day 60. 
Secondary 
Endpoint 10 
 
AE/SAE 
monitoring 
Adverse events, 
Continuously, 
from baseline 
to end of the 
study. 
Secondary 
Endpoint 11 
 
Tolerability 
 
- 
Secondary 
Endpoint 12 
 
Compliance 
- 
Sample Size 
A total of 29 participants will be enrolled, allowing for an anticipated 20% 
dropout rate to ensure at least 23 evaluable participants complete the study. 
Participant recruitment targets should strive for balance in sex (ideally, 50% 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 7 of 46 
female, but 40-60% balance is acceptable with accommodation for intersex 
individuals) 
Inclusion  
criteria 
 
Participants meeting all of the following criteria will be eligible for the 
study: 
1. Participants with  
• Generally good health with life expectance ≥5 years;  
• Male and female participants aged between 50-80 years having a BMI 
between 25–35 kg/m² (both inclusive); 
• Health not severely compromised (i.e. no life-threatening illness or 
disability) 
2. Participants with  
• Intrinsic Capacity score showing mild-to-moderate decline, and/or  
• Short Physical Performance Battery (SPPB) score between 9–11 and/or 
• Montreal Cognitive ≥ Assessment (MoCA) >20 and ≤25; 
3. Participants with or without comorbidity. If comorbidity is present, it 
should be on a stable regimen (i.e., same drug and same dose) for at least 
12 weeks before the screening visit; 
4. Participants are able and willing to provide written informed consent, 
comply with the study protocol requirements, and can read and write in 
English.  
Exclusion  
criteria 
Participants meeting any of the following criteria will not be eligible for the 
study:  
1. Severe or uncontrolled chronic disease (osteoarthritis, advanced 
cardiovascular disease, kidney failure, uncontrolled diabetes, severe 
COPD, terminal cancer); 
2. Physical disability requiring a walker or wheelchair; 
3. Diagnosed dementia or cognitive impairment preventing protocol 
compliance; 
4. Acute infection or illness within 3 months before enrolment; 
5. Unstable medical conditions (e.g., recent MI, unstable angina, 
uncontrolled hypertension); 
6. Major surgery in the past 6 months or planned during study; 
7. Severe psychiatric disorders unless well controlled; 
8. Substance abuse within past 6 months; 
9. Participation in another clinical trial within the last 6 months; 
10. Known allergy to any component of the nutraceutical kit; 
11. Participants with sedentary lifestyles are not able or not advised to 
undergo the resistance training as per protocol; 
12. Any clinically relevant macro or micro-nutrient deficiency as per 
investigator discretion; 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 8 of 46 
13. Pregnant women, lactating women, women of child bearing potential 
not following adequate contraceptive measures, women who were 
found positive for urine pregnancy test; 
14. Participants who are currently taking any herbal products, 
nutraceuticals, Ayurvedic formulations, dietary supplements, or 
vitamin/mineral supplements and are unwilling to discontinue their 
use for the duration of the study; 
15. According to the investigator, any other illness or abnormal laboratory 
investigations would interfere directly with the study results or 
jeopardize the participant's health. 
Study Duration 
Each patient will participate in this study for approximately 60 days 
(minimum). 
Study Visits 
Scheduled study visits: 
1. Screening visit (-7 to day 0) 
2. Visit 1: Baseline Visit: Enrolment (Day 1) 
3. Visit 2: Day 30 ± 5 days  
4. Visit 3: Day 60 ± 5 days (End of the study) 
End of the Study The end of the study will be defined if a patient meets the following criteria: 
1. Completer of the study 
2. Progression of disease, non-compliant patient, and as per discretion of  
investigator (Termination from the study) 
3. Incidence of SAE (Termination from the study) 
4. Patient withdraws consent  
5. Development of a new comorbidity during the study period, leading to 
discontinuation based on the investigator’s clinical judgment 
6. Depends upon the investigator’s discretion  
Interim Analysis There would be at least one interim analysis. It will be documented in the 
interim analysis record. 
Statistics 
All statistical analyses will be performed using IBM SPSS Statistics 
(Version 30) or an equivalent validated statistical software package. All 
statistical tests will be two-sided with a significance level (α) of 0.05. Given 
the exploratory, proof-of-concept nature of this single-arm pilot study, the 
emphasis will be on estimation of effect sizes and confidence intervals, with 
hypothesis testing considered supportive and descriptive. 
Continuous variables will be summarized using descriptive statistics: 
number of observations (n), mean, standard deviation (SD), median, 
minimum, and maximum. Categorical variables will be summarized using 
frequency counts and percentages. 
Patient 
Population 
for 
Analyses 
Enrolled Population 
All participants who sign informed consent and are enrolled into the 
study. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 9 of 46 
 
Safety Population (mITT) 
All enrolled participants who receive at least one dose of the 
investigational product. This population will be used for all safety and 
tolerability analyses, and for compliance assessments. 
Per-Protocol (PP) Population 
All participants in the Safety Population who: 
o complete the Day 60 visit (within the allowed window), 
o have primary endpoint data at both baseline and Day 60, and 
o have no major protocol deviations that, in the opinion of the 
investigator and/or statistician, may significantly impact primary 
efficacy outcomes, and 
o demonstrate treatment compliance of ≥90% (see compliance 
definition in Section X.7). 
The primary efficacy analyses will be based on the PP Population, with 
sensitivity analyses performed on the Safety (mITT) Population where 
applicable. Safety analyses will be based on the Safety Population. 
Participants who are enrolled but do not receive any dose of the 
investigational product will be listed separately under the category 
“Treatment not given” and will not be included in efficacy or safety 
summaries. 
Efficacy 
Analyses 
The efficacy endpoints will be analyzed using appropriate statistical tests, 
including the student’s paired or unpaired t-test, Wilcoxon signed-rank test, 
Mann–Whitney U test, or Chi-square test, depending on the nature of the 
data (parametric or non-parametric). 
The final statistical analysis plan (SAP) will be drafted after the locking of 
the database and will be documented and notified to the sponsor as well as 
ethics committees (IRBs).  
Safety Analysis 
Adverse events (AEs) and serious adverse events (SAEs) will be 
summarized by reporting both the total number of events and the number 
of participants experiencing at least one event during the study period. 
Additionally, these summaries will be stratified based on seriousness, 
severity, and their relationship to the study medication. The percentage of 
participants experiencing events and overall tolerability will be analyzed 
and compared using the Chi-square test.   
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 10 of 46 
6. INTRODUCTION 
6.1. Background information 
Humanity is experiencing a demographic shift unprecedented in history. By 2050, the number of 
people aged ≥60 years will surpass 2 billion, reshaping the landscape of healthcare, productivity, 
and social stability [1]. Although modern medicine has successfully extended lifespan, it has 
achieved far less in extending healthspan—the period of life lived with autonomy, mobility, 
cognitive vitality, and immune resilience. Aging is increasingly understood as a biological 
phenomenon characterized by metabolic inefficiency, mitochondrial decline, chronic 
inflammation, loss of neuromuscular mass, and immune dysregulation [2,3]. These interconnected 
processes erode the capacity of older adults to live actively and independently long before the 
onset of overt disease. 
Sarcopenia is a central contributor to late-life frailty, affecting 10–20% of individuals over 60 
years [4]. Progressive decreases in mitochondrial ATP output, muscle fiber density, and motor unit 
recruitment drive reductions in physical capability and gait endurance [3,6]. Functional 
performance measures such as the six-minute walk distance (6MWD) provide a holistic 
assessment of biological capacity—capturing cardiopulmonary endurance, neuromotor 
synchronization, musculoskeletal output, and motivation [7]. Clinically meaningful improvements 
in 6MWD (20–50 meters) are associated with reduced hospitalization, better survival, and delayed 
disability in older adults [7]. Thus, 6MWD embodies the real-world essence of healthspan better 
than isolated laboratory biomarkers. 
The European Working Group on Sarcopenia in Older People (EWGSOP2) specifically 
recommends DXA for confirming sarcopenia diagnosis and utilizes standardized cut-off points, 
such as ASM/height2 <7.0kg/m2 for men and <5.5kg/m2 for women, to identify clinically 
significant low muscle quantity [4]. The Dual-energy X-ray absorptiometry (DXA) system 
provides appendicular Lean Mass (ALM), which is widely accepted in research as a validated 
surrogate for appendicular skeletal muscle (ASM) mass and forms the foundation in all major 
sarcopenia consensus definitions as a clinically recognized research tool [4,5].  
Cognitive decline represents a second pillar of aging. Mild cognitive impairment and executive 
dysfunction emerge gradually, shaped by neuroinflammation, oxidative stress, and impaired 
neurogenesis [8]. Multifactorial nutraceuticals targeting mitochondrial function, synaptic 
plasticity, and antioxidative defense have demonstrated potential to improve cognitive 
performance in domains such as processing speed, working memory, and executive decision-
making [8]. Tools like the NIH Toolbox Fluid Composite enable comprehensive assessment of 
multiple functional domains over short time frames, reducing bias inherent in single-domain tests. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 11 of 46 
Aging also drives profound immune remodeling. Immunosenescence—a hallmark of aging—
encompasses reductions in naïve T-cell pools, impaired antigen presentation, and chronic low-
grade inflammation (“inflammaging”) [9]. Dysregulated cytokine networks such as elevated IL-6 
and TNF-α further propagate oxidative stress and tissue degeneration [9]. Nutritional and 
phytochemical interventions have demonstrated immunomodulatory activity, increasing NK cell 
surveillance and supporting adaptive immune balance [10]. 
The proposed nutraceutical combination aims to simultaneously enhance muscle function, 
cognitive performance, and immune resilience using synergistic bioactive components. Unlike 
single-target pharmaceuticals, nutraceutical combinations may modulate multiple biological 
axes—mitochondrial respiration, oxidative stress pathways, neurotrophic signaling, and immune 
regulation—providing versatility and scalability [2,10,11]. Most importantly, the intervention is 
accessible, non-invasive, and feasible for mass deployment across diverse socioeconomic 
contexts. 
If a short-term intervention can produce measurable improvements in functional mobility, 
cognitive performance, and immune resilience, it will demonstrate that aging is not a passive, 
unidirectional decline but a modifiable biological state. This work directly advances to extend the 
period of life lived in capability, autonomy, and human dignity. 
6.2. Details of Investigational Product 
6.2.1 Ingredients of Investigational Products 
     Table 3: Active Ingredients of Investigational product  
S/N 
Ingredient 
Composition (g) 
1 
Whey isolate 90% 
17.50 
2 
Vegan Collagen peptide 
4.00 
3 
Mango freeze dried 
5.00 
4 
Creatine monohydrate 
1.50 
5 
Magnesium glycinate 
0.50 
6 
Resveratrol 98% 
0.25 
7 
Quercetin 95% 
0.25 
8 
Cordyceps extract 
0.14 
9 
Marine algae (B12 + D3) 
0.10 
10 
Acetyl-L-Carnitine 
0.10 
11 
Phosphatidylserine 
0.10 
12 
AKBA 
0.12 
13 
Brahmi extract 
0.19 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 12 of 46 
14 
Ginkgo biloba extract 
0.10 
15 
Ashwagandha extract 
0.07 
16 
Seaweed extract 
0.05 
17 
Curcumin 95% 
0.03 
TOTAL 30.00 
• Other ingredients: Q.S. 
 
6.2.2 Pharmacology of Product ingredients: 
The investigational product includes scientifically validated bioactive ingredients targeting key 
determinants of health span: skeletal muscle function, cognitive performance, immune 
resilience, and inflammatory regulation. These compounds act across mitochondrial 
metabolism, synaptic neurotransmission, antioxidant activity, neuromuscular repair, and 
immune signalling pathways. 
• Whey Protein Isolate (90%) 
Whey isolate is a rapidly absorbed complete protein rich in leucine, which activates the mTOR 
pathway and increases skeletal muscle protein synthesis in aging populations [12,13]. 
Supplementation promotes recovery from activity-induced microtrauma and supports preservation 
of lean muscle mass relevant to sarcopenia mitigation [12]. 
• Creatine Monohydrate 
Creatine increases intramuscular phosphocreatine levels, enabling faster ATP regeneration during 
muscular contraction [14]. In older adults, creatine supplementation combined with resistance 
exercise improves muscle strength, lean mass, and functional performance including walking 
capacity and chair-rise time [15]. 
• Vegan Collagen Peptides 
Collagen peptides stimulate fibroblast activity and extracellular matrix synthesis, improving 
tendon integrity and muscle–tendon force transmission [16]. Supplementation increases collagen 
turnover, reduces joint discomfort, and supports exercise recovery, indirectly improving mobility 
and gait performance [17]. 
• Cordyceps Extract 
Cordyceps sinensis bioactives upregulate mitochondrial ATP production, enhancing endurance 
capacity and oxygen utilization [18]. The polysaccharide fraction also enhances macrophage and 
NK-cell responses, contributing to immune resilience in aging cohorts [19]. 
• Marine Algae (Vitamin D3 + Vitamin B12) 
Vitamin D3 modulates calcium handling and skeletal muscle contractility and is essential for 
immune homeostasis and T-cell activation [20]. Vitamin B12 supports neuronal myelination and 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 13 of 46 
neurotransmitter synthesis; deficiencies are associated with cognitive impairment, fatigue, and 
reduced neuroperformance [21]. 
• Acetyl-L-Carnitine (ALCAR) 
ALCAR facilitates mitochondrial transport of long-chain fatty acids, improving ATP production 
in both neurons and skeletal muscle [22]. Clinically, it demonstrates cognitive benefits such as 
improved mental energy, memory, and executive function in aging adults [23]. 
• Phosphatidylserine 
Phosphatidylserine is a core neuronal membrane phospholipid that enhances synaptic plasticity, 
promotes acetylcholine turnover, and reduces stress-induced cortisol elevations [24]. 
Supplementation improves accuracy, attention, and working memory performance [25]. 
• AKBA (Boswellia serrata) 
Acetyl-11-keto-β-boswellic acid (AKBA) is a selective 5-lipoxygenase inhibitor reducing 
leukotriene-mediated inflammation and oxidative tissue stress [26]. Clinical data demonstrates 
improved joint function and reduced inflammatory symptoms without NSAID-related 
gastrointestinal toxicity [27]. 
• Brahmi (Bacopa monnieri) 
Bacosides increase dendritic branching density, promote synaptic communication, and enhance 
retention of newly acquired information [28]. Chronic supplementation improves memory recall 
and reduces cognitive latency in aging cohorts [29]. 
• Ginkgo biloba 
Ginkgo flavone glycosides enhance cerebral blood flow, reduce oxidative neuronal damage, and 
modulate neurotransmitter activity [30]. It demonstrates clinically significant benefits in attention, 
psychometric performance, and age-related cognitive decline [31]. 
• Ashwagandha (Withania somnifera) 
Ashwagandha regulates the hypothalamic-pituitary-adrenal (HPA) axis and reduces cortisol, 
improving resilience, sleep, and cognitive performance [32]. It also enhances muscle strength and 
immune activation via NK-cell and macrophage pathways [33]. 
• Seaweed Extract (Fucoidans) 
Fucoidan polysaccharides exhibit immunomodulatory actions through macrophage stimulation, 
enhanced NK cytotoxicity, and antioxidant protection [34]. They have demonstrated anti-fatigue, 
anti-glycation, and metabolic stability effects relevant to aging [35]. 
• Curcumin (95%) 
Curcumin suppresses NF-κB, COX-2, and pro-inflammatory cytokine activity (TNF-α/IL-6), 
reducing systemic inflammation and oxidative stress [36]. It also demonstrates neuroprotective 
properties, supporting mitochondrial function and attenuating amyloid pathology [37]. 
• Mango Freeze-Dried (Polyphenols, Mangiferin) 
Mango pulp delivers mangiferin, xanthones, and flavonoids with potent antioxidant and anti-
inflammatory effects, reducing lipid peroxidation and reactive oxygen species burden relevant to 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 14 of 46 
aging physiology [38]. Mangiferin modulates glucose homeostasis, protects neuronal membranes, 
and demonstrates neurocognitive benefits by attenuating pro-oxidant cytokine cascades in 
preclinical and early translational models [39]. 
• Magnesium Glycinate 
Magnesium acts as a cofactor for more than 300 enzymatic reactions, including ATP synthesis, 
nerve conduction, and muscle contraction, and deficiency contributes to fatigue, impaired 
performance, and neuromuscular instability [40]. The chelated glycinate form offers superior 
gastrointestinal tolerance and supports sleep regulation, immune homeostasis, and neurocognitive 
stability through modulation of NMDA receptor excitability [41]. 
• Resveratrol 98% 
Resveratrol activates SIRT1 and AMPK signalling, promoting mitochondrial biogenesis, 
metabolic resilience, and longevity-associated pathways [42]. It exerts anti-inflammatory and 
antioxidant effects by downregulating NF-κB and inhibiting TNF-α/IL-6 expression, supporting 
immune and neurovascular integrity in aging populations [43]. 
• Quercetin 95% 
Quercetin is a flavonol with strong free-radical scavenging capacity, inhibiting xanthine oxidase 
and suppressing NF-κB–driven inflammatory cascades [44]. It modulates innate immunity through 
mast-cell stabilization and enhances antiviral and NK-cell responses, while also providing 
neuroprotection against oxidative injury [45]. 
 
6.2.3 Indications  
Improvement in muscle domain, immunity parameter and cognitive parameters in 
aging population. 
 
6.2.4 Dosage 
30 g sachet content to be mixed in water and consumed twice daily before meals. 
 
6.2.5 Packaging for Clinical Trial Supply 
30g sachet for each serving 
 
6.2.6 Storage Condition 
Store in a cool & dry place away from sunlight. Once opened the sachet needs to be 
consumed at once, no storage after opening of sachet. 
 
6.2.7 Shelf Life 
18 months from the date of manufacturing 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 15 of 46 
6.3. Preclinical and Clinical Study Data 
The investigational nutraceutical formulation integrates bioactive compounds with validated 
molecular actions on muscle physiology, cognitive performance, immune modulation, and 
systemic inflammation. The combination approach is grounded in the principle that aging is a 
multisystem decline requiring coordinated biological modulation rather than single-pathway 
intervention. Selected ingredients within the formula possess established clinical evidence across 
performance, neurocognitive, and immunological domains, which collectively support the 
rationale for conducting this proof-of-concept study. 
Muscle Function and Physical Performance 
Creatine monohydrate has undergone extensive clinical evaluation across aging and sarcopenic 
populations. Supplementation increases intramuscular phosphocreatine, enhances ATP 
regeneration, and augments resistance training outcomes, particularly lower-limb power and gait 
endurance [46]. Older adults receiving creatine in combination with strength training demonstrated 
superior improvements in lean mass and muscle performance compared to training alone [47]. 
Similarly, whey protein isolate, rich in leucine, stimulates mTORC1-dependent muscle protein 
synthesis and supports post-exercise recovery and anabolic maintenance in aging adults [48]. A 
synergistic interaction has been described whereby creatine enhances the anabolic sensitivity of 
muscle protein synthesis under protein-supplemented conditions [49]. 
Cognitive Performance and Neuroprotection 
Bacopa monnieri (Brahmi) has been repeatedly evaluated in aging populations; standardized 
extracts have shown improvements in memory acquisition, retention, and recall latency following 
chronic supplementation periods of 8–12 weeks [50]. The bacoside fraction promotes dendritic 
arborization and cholinergic neurotransmission, mechanisms associated with enhancement of 
working memory and attention. Ginkgo biloba has demonstrated benefits in cognitive processing 
speed and attention, particularly in subjects with age-related cognitive decline. Randomized trials 
suggest vascular-neuroprotective effects mediated through antioxidant flavonol fractions and 
improved cerebral perfusion [51]. Acetyl-L-Carnitine also exhibits neurotrophic activity, 
enhancing mitochondrial fatty acid transport and acetylcholine synthesis, with demonstrated 
improvements in cognitive symptoms in elderly cohorts [52]. 
Immune Function and Inflammation Modulation 
Cordyceps sinensis polysaccharides increase innate immune activation, promoting NK-cell 
cytotoxicity and macrophage functional responsiveness [53]. Clinical and preclinical findings 
describe improvements in exercise tolerance and VO₂max, likely mediated by mitochondrial ATP 
enhancement and oxidative stress reduction. Ashwagandha (Withania somnifera) exerts 
immunomodulatory and anti-stress properties; standardized extract administration reduces 
cortisol, enhances NK-cell activity, and improves subjective resilience scores in stressed adult 
populations [54]. Polyphenolic compounds, including quercetin and resveratrol, demonstrate 
potent NF-κB inhibition and attenuation of IL-6 and TNF-α expression, supporting mechanisms 
of inflammation reduction and mitochondrial protection in aging tissues [55,56]. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 16 of 46 
6.4. Risk-Benefit Assessment  
The investigational nutraceutical consists of widely used dietary ingredients with established 
human safety at recommended doses. Expected risks are low and generally limited to mild 
gastrointestinal discomfort, headache, or hypersensitivity in susceptible individuals [57,58]. 
Potential benefits include improvements in physical function, cognitive performance, immune 
balance, and inflammatory control, all of which are clinically relevant to older adults. With 
appropriate eligibility screening and ongoing safety monitoring, the anticipated benefits to 
participants are considered to outweigh the potential risks. 
7. STUDY RATIONALE  
The collective evidence indicates that the key constituents target distinct yet interconnected 
physiologic domains—mitochondrial ATP generation, neuromuscular anabolism, immune 
regulation, and oxidative stress. Unlike pharmaceutical monotherapies, dietary bioactives 
exhibit complementary mechanisms, enabling multi-domain functional improvement through 
moderate biological modulation. While each ingredient has been individually studied, no prior 
clinical investigation has systematically tested a combined formulation that concurrently 
addresses muscle endurance, cognition, immune resilience, and inflammation within a short-
duration intervention window. The current trial is therefore intended as a proof-of-concept to 
assess functional responses and biomarker trajectories in older adults.  
8. STUDY OBJECTIVES AND ENDPOINTS 
8.1. Study Objectives 
The primary objective of this study is to evaluate the impact of the combination nutraceutical 
on muscle function, as reflected by changes in performance and functional capacity. 
Secondary objectives include assessing its effects on cognitive performance, immune 
resilience, and inflammatory biomarkers, along with evaluating tolerability, safety, and 
participant compliance over the intervention period. This proof-of-concept trial aims to 
generate early clinical evidence supporting the formulation’s multi-domain benefits in older 
adults. 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 17 of 46 
8.2. Endpoints 
8.2.1. Efficacy endpoints 
Table 4: Efficacy Endpoints 
Domain 
Test and Method 
Outcome Measure 
Time frame 
Primary 
Endpoint 1 
 
6-minute walk test 
6-minute walk distance 
(6MWD) (meters) 
Screening, and 
Day 60. 
Secondary 
Endpoint 1 
 
1-RM strength test 
(knee extension) 
1 RM (kg) by leg press 
Screening, and 
Day 60. 
Secondary 
Endpoint 2 
 
DXA scan  
Muscle volume 
Screening, and 
Day 60. 
Secondary 
Endpoint 3 
 
NIH Toolbox Fluid 
Composite 
Fluid Composite score 
Screening, and 
Day 60. 
executive function, attention 
and processing speed, 
working memory 
Secondary 
Endpoint 4 
 
Cognitive 
Biomarkers 
p-tau181 
Screening, and 
Day 60. 
Secondary 
Endpoint 5 
 
Immunological 
markers  
CD4/CD8 ratio, CD45, 
CD3, CD4, CD8, T cells, B 
cells, NK cells, 
lymphocyte/neutrophil ratio 
Screening, and 
Day 60. 
Secondary 
Endpoint 6 
 
Inflammatory 
markers 
hs-CRP, TNF-alpha, IL-6 
Screening, and 
Day 60. 
Secondary 
Endpoint 7 
 
Longevity and 
Putative Energy 
Markers  
MDA, SOD, NAD+ 
Screening, and 
Day 60. 
Secondary 
Endpoint 8 
 
Anthropometric 
parameters 
Body Weight (kg), BMI 
(Kg/m2) and hip, waist, 
chest circumferences (cm) 
Screening, and 
Day 60. 
Secondary 
Endpoint 9 
 
Hematological & 
biochemical 
investigations  
Complete blood count 
(CBC), Renal Function Test 
(RFT). Liver Function Test 
(LFT), Thyroid profile, lipid 
profile, electrolytes. 
Screening, and 
Day 60. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 18 of 46 
Secondary 
Endpoint 10 
 
AE/SAE 
monitoring 
Adverse events, 
Continuously, 
from baseline to 
end of the study. 
Secondary 
Endpoint 11 
 
Tolerability 
 
- 
 
Secondary 
Endpoint 12 
 
Compliance 
- 
 
9. INVESTIGATIONAL PLAN  
9.1. Study Design 
This open-label, single arm interventional clinical study aims to evaluate the efficacy and 
safety of nutraceutical formulation (Interventional product) in older aduts. A total of 29 
participants will be screened and enrolled to ensure at least 23 evaluable cases. The total 
treatment period will be 60 days. 
9.2. Overall Study Plan 
This is an open-label, single-arm interventional clinical study designed to evaluate the 
efficacy, safety, and tolerability of the investigational nutraceutical formulation in older adults 
over a 60-day treatment period. A total of 29 participants will be screened, with the intent to 
obtain at least 23 evaluable subjects. Eligible participants will complete baseline assessments 
at screening and day 1, followed by administration of the study product for 60 days. The 
primary efficacy endpoint is improvement in functional capacity as assessed by change in 6-
minute walk distance from baseline to Day 60. Secondary endpoints include changes in 
muscle strength (1-RM knee extension), muscle volume (DXA), cognitive performance (NIH 
Toolbox Fluid Composite and p-tau181), immune cell profiles, inflammatory biomarkers (hs-
CRP, TNF-α, IL-6), oxidative and longevity-related markers (MDA, SOD, NAD+), 
anthropometry. Hematological and biochemical safety laboratories (CBC, RFT, LFT, thyroid 
profile, lipid profile, electrolytes) will be evaluated at baseline and Day 60. Adverse events, 
serious adverse events, tolerability, and compliance will be monitored continuously from 
enrollment to end of study. Participants will attend a screening visit (−7 to 0 days), 
baseline/enrollment visit (Day 1), and an end-of-study visit on Day 60 ± 5 days. 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 19 of 46 
Diagram depicting the overall study plan: 
 
9.3. Selection and Handling of Study Population 
The research team will recruit more than 23 eligible adult participants (both men and women) 
at the study site. 
9.3.1.  Inclusion Criteria 
        Participants will be eligible for the study if they meet all of the following conditions: 
1. Participants with  
• Generally good health with life expectance ≥5 years;  

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 20 of 46 
• Male and female participants aged between 50-80 years having a BMI between 25–35 
kg/m² (both inclusive); 
• Health not severely compromised (i.e. no life-threatening illness or disability) 
2. Participants with  
• Intrinsic Capacity score showing mild-to-moderate decline, and/or  
• Short Physical Performance Battery (SPPB) score between 9–11 and/or 
• Montreal Cognitive ≥ Assessment (MoCA) >20 and ≤25; 
3. Participants with or without comorbidity. If comorbidity is present, it should be on a stable 
regimen (i.e., same drug and same dose) for at least 12 weeks before the screening visit; 
4. Participants are able and willing to provide written informed consent, comply with the study 
protocol requirements, and can read and write in English. 
 
9.3.2. Exclusion criteria 
Participants meeting any of the following criteria will not be eligible for the study:  
1. Severe or uncontrolled chronic disease (osteoarthritis, advanced cardiovascular disease, 
kidney failure, uncontrolled diabetes, severe COPD, terminal cancer); 
2. Physical disability requiring a walker or wheelchair; 
3. Diagnosed dementia or cognitive impairment preventing protocol compliance; 
4. Acute infection or illness within 3 months before enrolment; 
5. Unstable medical conditions (e.g., recent MI, unstable angina, uncontrolled hypertension); 
6. Major surgery in the past 6 months or planned during study; 
7. Severe psychiatric disorders unless well controlled; 
8. Substance abuse within past 6 months; 
9. Participation in another clinical trial within the last 6 months; 
10. Known allergy to any component of the nutraceutical kit; 
11. Participants with sedentary lifestyles are not able or not advised to undergo the resistance 
training as per protocol; 
12. Any clinically relevant macro or micro-nutrient deficiency as per investigator discretion; 
13. Pregnant women, lactating women, women of child bearing potential not following adequate 
contraceptive measures, women who were found positive for urine pregnancy test; 
14. Participants who are currently taking any herbal products, nutraceuticals, Ayurvedic 
formulations, dietary supplements, or vitamin/mineral supplements and are unwilling to 
discontinue their use for the duration of the study; 
15. According to the investigator, any other illness or abnormal laboratory investigations would 
interfere directly with the study results or jeopardize the participant's health. 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 21 of 46 
9.3.3. Discontinuation Criteria 
Participants will be considered as discontinued based on the following criteria: 
• Withdrawal of consent by the participant to continue in the study; 
• The appearance of non-tolerable adverse events; 
• The appearance of exclusion criteria, which concern the safety of the participant; 
• Any situation judged by the investigator to be harmful to the participant;  
• At the discretion of the investigator, where the clinical condition of the participant worsens; 
• At the discretion of the investigator, poor compliance of the participant to study procedures; 
• A positive pregnancy test at any time during the study period; 
• Protocol deviation that in the opinion of the sponsor and investigator warrants discontinuation 
from the study; 
• The participant suffered from inter-current illness during the study. 
The reason for participant discontinuation will be documented in the participant’s case report 
form (CRF). Discontinued number of participants will not be replaced. In case of premature 
discontinuation, all follow-up assessments will be conducted at early withdrawal. Participants 
discontinued from the study at any stage will be considered for safety analysis.  
9.4. End of Study  
The end of the study will be defined if a participant meets the following criteria: 
1. Completer of the study. 
2. Progression of disease, non-compliant participant, and as per discretion of investigator. 
3. Incidence of SAE (Termination from the study). 
4. The participant withdraws consent. 
5. Depends upon the investigator's discretion. 
9.5. Blinding 
The study is open label study, no need of blinding process. 
9.6. Handling of Investigational Product 
Supply and Receipt 
The study IPs will be labeled according to the participant number before the start of the study. The 
investigator will be overall responsible for ensuring that IP is stored in a safe, limited access 
location, as per the condition noted on the label. 
The study IP will be labeled and packaged such that the label should include but not be limited to 
the Sponsor name/address, dosage, route of administration, name/strength, batch number/code, 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 22 of 46 
study reference or protocol number, directions for use, the statement ‘for clinical trial use only’, 
storage conditions, expiry/use by date.  
Study IP will be handled by designated personnel at the site. The study personnel involved in IP 
handling will not participate in any activity pertaining to participant safety and efficacy assessment 
that may impact the study outcome. 
It is the responsibility of the investigator to designate independent dispensers or designated 
personnel to ensure that the current disposition of the IP is maintained at each study site where IP 
is inventoried and dispensed. When investigational product shipment is received at the study site, 
independent dispensers or designated personnel must acknowledge receipt of IP (An attempt 
would be made to store the IP at the required temperature as early as possible). 
Accountability and Retentions of Investigational Product 
Independent dispensers or designated study personnel will maintain accurate records in an IP 
accountability log of receipt of all IP, inventory at the site, dispensing of IP, disposition of used 
products, and return of unused products to the sponsor. The study IP (used/unused) will be 
disposed of at the site with proper investigational product accountability maintained. The IP 
disposition report will be generated separately at the close out of the site.  
Dosage and Administration of IPs 
30g sachet content will be mixed in water and consumed twice a day before meals. 
Treatment Compliance 
Treatment compliance of each participant concerning the administration of the IP will be recorded 
in the respective section of the CRF. The site will maintain the accountability of all used and 
unused IP. The cut-off for non-compliance will be a maximum of up to 10% per month. If any 
participant misses doses for more than permissible, it would be considered a dropout.  
9.7. Clinical Procedures and Assessments 
The following are the clinical procedures and assessments during the study period.  
Screening Assessments (Day -7 to Day 0) 
Screening procedures will be performed within 7 days before randomization. All aspects of the 
study will be explained to the participants at screening. The investigator must ensure that the 
participant meets all of the inclusion and none of the exclusion criteria. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 23 of 46 
The following activities will be performed before randomization to screen the participants: 
• 
Presentation of participant information and obtaining participant’s written informed 
consent 
• 
Recording of demographic data (age, gender) 
• 
Concomitant diseases/ concomitant medication 
• 
Inclusion/Exclusion criteria review 
• 
Vital parameters: 
- 
blood pressure, heart rate, body temperature, and respiratory rate 
• 
Physical examination 
• 
Clinical examination 
• 
Assessment of 6-minute walk distance (6MWD) (meters) 
• 
Assessment of 1RM (kg) with the leg press 
• 
Assessment of Muscle volume 
• 
Assessment of Fluid Composite score and assessment of executive function, 
attention and processing speed, working memory 
• 
Assessment of plasma p-tau181 
• 
Assessment of CD4/CD8 ratio, CD45, CD3, CD4, CD8, T cells, B cells, NK cells, 
lymphocyte/neutrophil ratio 
• 
Assessment of serum hs-CRP, TNF-alpha, IL-6 
• 
Assessment of plasma MDA and SOD, NAD+ from whole blood 
• 
Assessment of Body Weight (kg), BMI (Kg/m2) and hip, waist, chest 
circumferences (cm) 
• 
Assessment of complete blood count (CBC), Renal Function Test (RFT). Liver 
Function Test (LFT), Thyroid profile, lipid profile, electrolytes. 
 
Visit 1: Baseline/Randomization (Day 1) 
• 
Clinical examination 
• 
Physical examination 
• 
Randomization 
• 
Dispensing of IP 
• 
Vital Parameters  
- 
blood pressure, heart rate, body temperature, and respiratory rate. 
• 
Recording and assessment of adverse events  
• 
Assessment of rescue medications 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 24 of 46 
Visit 2: Day 30±5 days 
• 
Clinical examination 
• 
Physical examination 
• 
Vital Parameters  
- 
blood pressure, heart rate, body temperature and respiratory rate 
• 
Dispensing of IP 
• 
Assessment of tolerability of investigational product 
• 
Assessment of compliance 
• 
Recording and assessment of adverse events  
• 
Assessment of rescue medications 
 
Visit 3: Day 60±5 days (End of the study) 
• 
Clinical examination 
• 
Physical examination 
• 
Vital Parameters  
- 
blood pressure, heart rate, body temperature and respiratory rate 
• 
Assessment of tolerability of investigational product 
• 
Assessment of compliance 
• 
Recording and assessment of adverse events  
• 
Assessment of rescue medications 
• 
Assessment of 6-minute walk distance (6MWD) (meters) 
• 
Assessment of 1RM (kg) with the leg press 
• 
Assessment of Muscle volume 
• 
Assessment of Fluid Composite score and assessment of executive function, 
attention and processing speed, working memory 
• 
Assessment of plasma p-tau181 
• 
Assessment of CD4/CD8 ratio, CD45, CD3, CD4, CD8, T cells, B cells, NK cells, 
lymphocyte/neutrophil ratio 
• 
Assessment of serum hs-CRP, TNF-alpha, IL-6 
• 
Assessment of plasma MDA and SOD, NAD+ from whole blood 
• 
Assessment of Body Weight (kg), BMI (Kg/m2) and hip, waist, chest 
circumferences (cm) 
• 
Assessment of complete blood count (CBC), Renal Function Test (RFT). Liver 
Function Test (LFT), Thyroid profile, lipid profile, electrolytes. 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 25 of 46 
9.8. Prohibited Concomitant Therapy 
Participants will be instructed to refrain from the use of products that may confound the 
interpretation of efficacy, biomarker, or safety outcomes during the study period. The following 
therapies are prohibited for the duration of participation unless otherwise approved by the 
Investigator: 
• Nutraceuticals, dietary supplements, herbal preparations, or Ayurvedic formulations 
(including multivitamins, antioxidants, adaptogens, anti-inflammatory extracts, amino acids, 
or energy boosters). 
• Investigational medicinal products or participation in another clinical trial within the last 6 
months. 
• Anabolic agents or muscle-enhancing therapies, including testosterone boosters, or growth 
hormone derivatives. 
• Cognitive enhancement agents such as prescription nootropics, cholinesterase inhibitors, 
stimulants, or memory enhancement blends. 
• Immunomodulatory agents, including biologics, corticosteroids, or disease-modifying anti-
rheumatic drugs (DMARDs), except when medically necessary and stable for ≥12 weeks 
prior to screening. 
• High-dose vitamin or mineral supplementation (e.g., vitamin D >2000 IU/day, Vitamin B12 
injection, NAD+ infusions, IV antioxidant cocktails, etc.). 
• Special diets or extreme caloric restriction protocols (ketogenic, intermittent fasting ≥18 
hours/day, very low-calorie diets, medical detox regimens). 
 
The Investigator may discontinue a participant if prohibited therapies are initiated during the trial 
or if continued use could compromise safety, study integrity, or endpoint assessment. 
9.9. Permitted Concomitant and Rescue Therapy 
Stable and medically necessary therapies that are not expected to interfere with study endpoints 
may be continued during the study. These include: 
• Chronic medications for pre-existing stable conditions (e.g., hypertension, diabetes, 
dyslipidemia) provided that the drug and dose have remained unchanged for ≥12 weeks 
prior to screening. 
• Occasional use of standard analgesics (e.g., acetaminophen/paracetamol up to 2 g/day) for 
mild to moderate pain not related to study intervention. 
• Routine dietary intake, including normal balanced meals, caffeine intake, and hydration 
practices. 
• Short-term symptomatic treatment, such as local antiseptics or mild topical medications 
for non-study-related conditions. 
• Rescue therapy, at Investigator discretion, for clinically significant deterioration, 
intolerable symptoms, or medical emergencies. In such cases, the intervention type, 
dosage, and duration will be documented in the source data and CRF. 
All permitted concomitant medications, rescue therapies, and dose changes must be recorded, 
including start/stop dates and medical indications. The Principal Investigator will assess whether 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 26 of 46 
continuation or withdrawal from the study is appropriate if any therapy is expected to confound 
efficacy or safety outcomes. 
9.10. Protocol Deviations and Violations 
The investigator shall strictly adhere to the protocol approved by the regulatory authority and IRB. 
All protocol ‘violations’ (for inclusion/exclusion) and protocol ‘deviations’ (related to study 
procedures) must be reported to the study monitor and a protocol deviation/violation log should 
be filled for every such case. 
9.11. Study Duration 
Each participant will participate in this study for approximately 60 days. 
 
10. SAFETY CONSIDERATIONS 
Safety and tolerability of study medication will be assessed by reporting and analyzing the adverse 
events (AEs) that occurred, vital signs, etc. The investigator asks the participant generally about 
adverse events without indicating special symptoms.  
10.1 Risks and Benefits to Study Participants 
In this planned clinical study, the participants will be informed about the expected risks and the 
inconveniences associated with the clinical trial. In addition, in compliance with ICH-GCP, 
participants are free to leave the clinical trial at any time they wish and are treated according to 
standard supportive care. To follow the risks adequately the investigator will see the participants 
frequently according to the clinical trial scheme provided in the protocol. 
The clinical study protocol includes discontinuation criteria on participants and a clinical trial basis 
to reduce individual risks. 
10.2 Definitions Related to Safety 
10.2.1 Adverse Event 
An AE (or adverse experience) is any untoward medical occurrence in a participant or clinical 
investigation participant administered a pharmaceutical product and which does not necessarily 
have to have a causal relationship with this treatment. It can therefore be any unfavorable and 
unintended sign (including an abnormal laboratory finding, for example), symptom, or disease 
temporally associated with the use of a medicinal product, whether or not considered related to the 
medicinal product. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 27 of 46 
Also, abnormal results of diagnostic procedures are considered to be AE, if the abnormality results 
in participant withdrawal, is associated with SAE, leads to additional treatment or to further 
diagnostic tests, or is considered to be of clinical significance by the investigator.  
Any “untoward medical occurrence” occurring before the administration of the study medication, 
does not represent an adverse event, but is reported as a “baseline complaint”. 
 
Pre-existing medical conditions or symptoms occurring before the initiation of the study will not 
be reported as AEs. A worsening of a pre-existing medical condition or symptom will be reported 
as an AE. 
10.2.2 Adverse Drug Reaction 
In the pre-approval clinical experience with a new medicinal product or its new usages, 
[particularly as the therapeutic dose(s) may not be established], all noxious and unintended 
responses to a medicinal product related to any dose will be considered adverse drug reactions. 
The phrase "responses to medicinal products" means that a causal relationship between a medicinal 
product and an AE is at least a reasonable possibility, i.e., the relationship cannot be ruled out. 
Regarding marketed medicinal products, an adverse drug reaction in the post-marketing setting is 
a response to a drug that is noxious and unintended and which occurs at doses normally used in 
humans for prophylaxis, diagnosis, or therapy of disease or modification of physiological function. 
10.2.3 Unexpected Adverse Drug Reaction 
An adverse event or suspected adverse reaction is considered “unexpected” if it is not listed in the 
investigator brochure or is not listed at the specificity or severity that has been observed; or, if an 
investigator brochure is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current application, as amended. 
10.2.4 Change of Laboratory Parameters 
If, after administration of study medication, changes in pathological laboratory values occur, 
which were not present before the treatment started, further clinical or laboratory tests must be 
carried out until the values return to the normal range or until a plausible explanation is found (e.g. 
concomitant disease) for the change of the laboratory values. 
A change in laboratory parameters during the study represents an adverse event: 
- 
if the change is clinically relevant. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 28 of 46 
Pathological values still present at end of the study, which were not present before the start of the 
study, have to be followed up by further appropriate investigations until the values have returned 
to normal range or until an explanation (e.g. concomitant disease) has been found. 
10.2.5 Serious Adverse Event or Adverse Drug Reaction 
A SAE is any untoward medical occurrence that at any dose: 
▪ results in death; 
▪ permanent disability; 
▪ is life-threatening [The term "life-threatening" in the definition of "serious" refers to an event 
in which the participant was at risk of death at the time of the event; it does not refer to an 
event which hypothetically might have caused death if it were more severe; 
▪ requires inpatient hospitalization or prolongation of existing hospitalization; 
▪ results in persistent or significant disability/incapacity, or; 
▪ is a congenital anomaly/birth defect.  
Medical and scientific judgment will be exercised in deciding whether expedited reporting is 
appropriate in other situations, such as important medical events that may not be immediately life-
threatening or result in death or prolongation of hospitalization but may jeopardize the participant 
or may require intervention to prevent one of the other outcomes listed in the definition above. 
These will also usually be considered “serious” as well. 
10.3 Adverse Event Documentation and Reporting 
The documentation of AEs starts with randomization for each participant and ends with the 
discharge of the participant from the hospital. If an AE occurs after the termination of the clinical 
trial of the individual participant and if the investigator judges the AE to be at least possibly related 
to the clinical trial, the investigator has to inform the sponsor. 
All AE related, signs or symptoms that are reported will be recorded in the AE form as part of 
CRF. In particular, the information will include details of the administration of the investigational 
product and details of AE with its date, onset time, causality, frequency, severity, outcome, and if 
any treatment or diagnostic steps were taken in relation to it. All AEs will be followed until 
resolution. All the AEs will be evaluated based on severity, causality, and outcome as given in the 
below table. 
      Table 5: AE evaluation based on severity, causality, and outcome. 
 Parameter 
Description 
Severity 
Grade 1 
Mild; asymptomatic or mild symptoms; clinical or diagnostic 
observations only; intervention not indicated. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 29 of 46 
 Parameter 
Description 
Grade 2 
Moderate; minimal, local, or noninvasive intervention indicated; 
limiting age-appropriate instrumental activities of daily living (ADL)*. 
Grade 3 
Severe or medically significant but not immediately life-threatening; 
hospitalization or prolongation of hospitalization indicated; disabling; 
limiting self-care ADL**. 
Grade 4 
Life-threatening consequences; urgent intervention indicated. 
Grade 5 
Death related to AE. 
Causality  
Certain 
▪ Event or laboratory test abnormality, with plausible time 
relationship to drug intake. 
▪ Cannot be explained by disease or other drugs. 
▪ Response 
to 
withdrawal 
plausible 
(pharmacologically, 
pathologically). 
▪ Event definitive pharmacologically or phenomenologically (i.e. an 
objective and specific medical disorder or a recognized 
pharmacological phenomenon). 
▪ Rechallenge satisfactory, if necessary. 
Probable /  
Likely 
▪ Event or laboratory test abnormality, with reasonable time 
relationship to drug intake. 
▪ Unlikely to be attributed to disease or other drugs. 
▪ Response to withdrawal is clinically reasonable. 
▪ Rechallenge is not required. 
Possible 
▪ Event or laboratory test abnormality, with reasonable time 
relationship to drug intake. 
▪ Could also be explained by disease or other drugs. 
▪ Information on drug withdrawal may be lacking or unclear. 
Unlikely 
▪ Event or laboratory test abnormality, with a time to drug intake that 
makes a relationship improbable (but not impossible). 
▪ Disease or other drugs provide plausible explanations. 
Conditional/ 
Unclassified 
▪ Event or laboratory test abnormality. 
▪ More data for proper assessment is needed, or; 
▪ Additional data under examination. 
Unassessable/ 
Unclassifiable 
▪ A report suggesting an adverse reaction. 
▪ Cannot be judged because information is insufficient or 
contradictory. 
▪ Data cannot be supplemented or verified. 
Outcome 
Resolved / Resolved with sequel / Ongoing / Stable chronic condition / Continuing at 
death / Death 
*Instrumental ADL refers to preparing meals, shopping for groceries or clothes, using the 
telephone, managing money, etc. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 30 of 46 
**Self-care ADL refers to bathing, dressing, undressing, feeding self, using the toilet, taking 
medications, and not being bedridden. 
Any AE regardless of casualty relationship will be recorded and reported to the IRB. 
AEs will be coded as per the MedDRA current version. 
10.4 Reporting of Serious Adverse Event 
10.4.1 Central Licensing Authority Requirements for Expedited Reporting of SAE 
▪ SAE will be reported by the clinical investigator to the sponsor, chairperson of EC, safety 
review board chairman, and central licensing authority within 24 hours of the knowledge of its 
occurrence. 
▪ The clinical investigator and sponsor will submit a detailed report of SAE to the chairperson of 
EC, the head of the clinical trial site, and the central licensing authority within 14 days of the 
knowledge of its occurrence. 
▪ IEC will send its review report to the central licensing authority for SAE within 30 days of its 
occurrence.   
▪ In case the clinical investigator fails to report any SAE within the stipulated period, he shall 
furnish the reason for the delay to the satisfaction of the licensing authority along with the report 
of SAE. 
▪ If applicable, the central licensing authority and safety review board recommended 
compensation by order will be paid by the sponsor within 30 days of receipt of such order from 
the central licensing authority for the SAE that occurred. 
All SAEs will be followed until a satisfactory resolution or until the investigator deems the event 
to be chronic or the participant to be stable. Participants /LAR will inform the investigator 
regarding the SAEs (whether related or unrelated) within 24 hours of their occurrence. 
The details of the sponsor’s contact personnel for SAE reporting are given below: 
Dr. Shridhar Pandya 
gplifehealthcare@gmail.com 
 
10.4.2 Reporting of Pregnancy 
Pregnancy is not expected due to age range of participants (50–80 years). However, if pregnancy 
is identified at any time during the study or within 30 days after last administration of the 
investigational product, it will be reported to the Sponsor and the Ethics Committee as a safety 
event. The participant will be withdrawn from the study, and appropriate medical follow-up will 
be arranged. Pregnancy outcomes and fetal/neonatal health will be documented in accordance with 
institutional and regulatory requirements. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 31 of 46 
11. STATISTICAL CONSIDERATIONS AND EFFICACY EVALUATION 
11.1 Sample Size Consideration 
Base sample size (evaluable subjects) 
From G*Power: 
• 
Effect size (Cohen’s d) = 0.70 
• 
Power (1–β) = 90% 
• 
α = 0.05 
• 
Design: Paired t-test (pre–post change) 
• 
➤ Required = 23 evaluable subjects 
Adjust for 20% dropout 
 
Approximate Formula for Sample Size in Paired t-Test 
 
 
Simplified version using Cohen’s d: 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 32 of 46 
 
Approximately 22–23 participants are required. 
 
This single-arm pilot study aims to evaluate the effect of the investigational nutraceutical on muscle 
strength, measured as changes in one-repetition maximum (1RM) in elderly male participants. A relevant 
reference is the randomized trial conducted by Kim et al. (2023), which assessed resistance training 
intensity in elderly adults and reported statistically significant improvements in lower-limb strength and 
functional performance after 12 weeks. In that study, differences in 1RM gains across intervention arms 
corresponded to standardized effect sizes ranging from d = 0.60 to 0.75, depending on training intensity 
and muscle group. 
Based on this precedent, and in consideration of our study’s nutraceutical-based intervention without formal 
resistance training, a conservative within-subject effect size of d = 0.70 is considered appropriate for this 
pilot. Using G*Power 3.1.9.7 software for a paired-sample t-test with two-tailed α = 0.05, 90% power, 
and Cohen’s d = 0.70, the estimated sample size is 23 evaluable participants. 
To accommodate an expected 20% dropout rate, the study will enrol 29 participants. 
 
Reference: 
Kim S, et al. Effects of Resistance Training Intensity on Muscle Strength and Functional Performance in 
Elderly 
Adults: 
A 
Randomized 
Trial. 
Appl 
Sci. 
2023;15(2):757. 
https://www.mdpi.com/2076-3417/15/2/757 
 
11.2 Statistical analysis plan 
General Principles 
All statistical analyses will be performed using IBM SPSS Statistics (Version 30) or an equivalent 
validated statistical software package. All statistical tests will be two-sided with a significance 
level (α) of 0.05. Given the exploratory, proof-of-concept nature of this single-arm pilot study, the 
emphasis will be on estimation of effect sizes and confidence intervals, with hypothesis testing 
considered supportive and descriptive. 
Continuous variables will be summarized using descriptive statistics: number of observations (n), 
mean, standard deviation (SD), median, minimum, and maximum. Categorical variables will be 
summarized using frequency counts and percentages. 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 33 of 46 
Analysis Populations 
• 
Enrolled Population 
• 
All participants who sign informed consent and are enrolled into the study. 
• 
Safety Population (mITT) 
• 
All enrolled participants who receive at least one dose of the investigational product. This 
population will be used for all safety and tolerability analyses, and for compliance 
assessments. 
• 
Per-Protocol (PP) Population 
• 
All participants in the Safety Population who: 
o complete the Day 60 visit (within the allowed window), 
o have primary endpoint data at both baseline and Day 60, and 
o have no major protocol deviations that, in the opinion of the investigator and/or 
statistician, may significantly impact primary efficacy outcomes, and 
o demonstrate treatment compliance of ≥90% (see compliance definition in Section 
X.7). 
The primary efficacy analyses will be based on the PP Population, with sensitivity analyses 
performed on the Safety (mITT) Population where applicable. Safety analyses will be based on 
the Safety Population. 
Participants who are enrolled but do not receive any dose of the investigational product will be 
listed separately under the category “Treatment not given” and will not be included in efficacy or 
safety summaries. 
 
Definition of Endpoints 
Primary Efficacy Endpoint 
• 
Change in 6-minute walk distance (6MWD, meters) from baseline (Screening) to Day 
60. 
Key Secondary Efficacy Endpoints 
Changes from baseline to Day 60 in: 
1. Muscle Function 
o 1-RM knee extension strength: peak torque / power and rate of torque development 
o DXA-based muscle volume (pre-specified muscle group/region of interest) 
2. Cognitive Performance 
o NIH Toolbox Fluid Composite Score (executive function, attention/processing 
speed, working memory) 
o Cognitive biomarker: plasma p-tau181 
3. Immune and Inflammatory Function 
o Immunological markers: CD4/CD8 ratio, CD45, CD3, CD4, CD8, T cells, B cells, 
NK cells, lymphocyte/neutrophil ratio 
o Inflammatory markers: hs-CRP, TNF-α, IL-6 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 34 of 46 
4. Longevity / Energy / Oxidative Stress Markers 
o MDA, SOD, NAD⁺ 
5. Anthropometric and Body Composition Parameters 
o Body weight (kg), BMI (kg/m²), waist, hip, chest circumference (cm) 
6. Clinical Laboratory Parameters 
o Complete blood count (CBC) 
o Renal function tests (RFT) 
o Liver function tests (LFT) 
o Thyroid profile 
o Lipid profile 
o Electrolytes 
Safety and Tolerability Endpoints 
• 
Incidence, frequency, and severity of adverse events (AEs) and serious adverse events 
(SAEs). 
• 
Relationship of AEs/SAEs to study product (related / possibly related / not related). 
• 
Overall tolerability based on investigator and participant assessment (e.g., well tolerated / 
moderately tolerated / poorly tolerated). 
Compliance Endpoint 
• 
Compliance (%) will be calculated as: 
 
Descriptive statistics (mean, SD, median, range) will be provided for compliance. The number and 
percentage of participants with Compliance ≥90% will be reported. Only participants with 
Compliance ≥90% will be included in the PP efficacy analyses. 
 
Baseline Characteristics 
Baseline demographic and clinical characteristics (e.g., age, sex, BMI, comorbidities, baseline 
6MWD, SPPB, MoCA, intrinsic capacity score) will be summarized for: 
• 
Enrolled Population 
• 
Safety (mITT) Population 
• 
PP Population 
No formal statistical testing of baseline differences will be performed, as this is a single-arm study; 
summaries are descriptive and used to characterize the sample. 
 
Primary Efficacy Analysis 
The primary endpoint is the change in 6MWD (meters) from baseline to Day 60. 
1. Derivation of Change Score 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 35 of 46 
o For each participant, change in 6MWD = 6MWD at Day 60 − 6MWD at baseline 
(Screening). 
2. Descriptive Analysis 
o The change in 6MWD will be summarized using n, mean, SD, median, minimum, 
maximum, and 95% confidence interval (CI) for the mean change. 
3. Inferential Analysis 
o Normality of the change scores will be assessed using the Shapiro–Wilk test. 
o If approximately normally distributed, a paired Student’s t-test will be used to 
evaluate whether the mean change from baseline to Day 60 differs from zero. 
o If normality assumptions are not met, the Wilcoxon signed-rank test will be used 
instead. 
4. Responder Analysis (Exploratory Within Primary Endpoint) 
o A responder may be defined as a participant with an improvement in 6MWD of 
≥30 meters from baseline to Day 60. 
o The proportion of responders will be summarized descriptively (n, %) with 95% 
CIs (e.g., Clopper–Pearson) and may be used for exploratory interpretation of 
clinically meaningful benefit. 
The primary analysis set for this endpoint will be the PP Population. 
 
Secondary and Exploratory Efficacy Analyses 
All secondary and exploratory endpoints will be analyzed in a similar manner, focusing on within-
subject change from baseline to Day 60. 
Continuous Secondary Endpoints 
For each continuous secondary endpoint (e.g., 1-RM strength, DXA based muscle volume, NIH 
Fluid Composite, biomarkers, anthropometrics, laboratory measures): 
1. Change Score 
o Change = value at Day 60 − value at baseline. 
2. Descriptive Summary 
o n, mean, SD, median, minimum, maximum, and 95% CI for the mean change. 
3. Inferential Testing 
o Normality assessed via Shapiro–Wilk test. 
o If normal: paired t-test for change from baseline. 
o If non-normal: Wilcoxon signed-rank test. 
4. Data Transformation (if needed) 
o Skewed biomarkers (e.g., hs-CRP, IL-6, TNF-α, MDA) may be log-transformed 
prior to analysis. In such cases, analyses will be based on transformed values, and 
descriptive statistics may be presented for both raw and transformed data as 
appropriate. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 36 of 46 
No multiplicity adjustment will be applied; these analyses are considered supportive and 
exploratory. 
Correlation and Exploratory Relationship Analyses 
Exploratory analyses may be performed to evaluate associations such as: 
• 
Relationship between change in 6MWD and changes in: 
o NIH Fluid Composite score 
o muscle strength and muscle volume 
o immune and inflammatory markers 
o oxidative stress and NAD⁺ levels 
Depending on data distribution: 
• 
Pearson correlation for approximately normal variables 
• 
Spearman rank correlation for non-normal variables 
Simple linear regression models may be used exploratorily to assess whether changes in selected 
biomarkers or strength measures predict change in 6MWD. These analyses will be considered 
exploratory and hypothesis-generating only. 
 
Safety, Tolerability, and Compliance Analyses 
Adverse Events 
All AEs and SAEs reported by participants or observed by investigators from first dose until end 
of study will be coded using a standard terminology (e.g., MedDRA, if applicable). 
AEs/SAEs will be summarized by: 
• 
number of events 
• 
number and percentage of participants experiencing at least one event 
• 
severity (mild, moderate, severe) 
• 
seriousness (serious / non-serious) 
• 
relationship to investigational product (related / possibly related / not related) 
Summaries will be presented for the Safety (mITT) Population. No formal hypothesis testing is 
planned; however, descriptive comparisons of proportions (e.g., between tolerability categories) 
may be conducted using Chi-square test or Fisher’s exact test, where appropriate. 
 
Clinical Laboratory Parameters, Vital Signs, and Other Safety Parameters 
Continuous safety-related variables (e.g., hematology, biochemistry, thyroid, lipid profile, 
electrolytes) will be summarized at baseline and Day 60, and change from baseline will be 
described using descriptive statistics. Potentially clinically significant abnormalities will be 
flagged and listed. 
 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 37 of 46 
Compliance 
Descriptive statistics (mean, SD, median, range) will be provided for compliance. The number and 
percentage of participants with Compliance ≥90% will be reported. Only participants with 
Compliance ≥90% will be included in the PP efficacy analyses. 
 
Handling of Missing Data 
No formal imputation of missing data is planned for primary analysis. Analyses will generally be 
based on observed cases: 
• 
Participants with missing baseline or Day 60 values for the primary endpoint will not 
contribute to the respective change analysis in the PP Population. 
• 
For sensitivity purposes, where appropriate, Last Observation Carried Forward (LOCF) 
may be used for participants who discontinue after at least one post-baseline assessment; 
such analyses will be clearly labeled as sensitivity analyses. 
The extent and pattern of missing data will be summarized descriptively and may be explored to 
assess whether missingness appears random or associated with baseline characteristics. 
 
Interim Analysis 
A single interim review may be conducted after a subset of participants completes the Day 30 visit. 
The purpose of this interim review will be to evaluate: 
• 
feasibility and recruitment 
• 
protocol adherence and data quality 
• 
safety signals and tolerability 
• 
variability of primary and secondary endpoints endpoint  
During the interim analysis, the primary and secondary endpoints may be analyzed using the same 
descriptive and within-subject change approach (paired t-test or Wilcoxon) to assess trends. These analyses 
will be exploratory only, without inferential conclusions or decision-making. 
 
Statistical Analysis Plan Finalization 
A more detailed internal Statistical Analysis Plan (SAP), if required, may elaborate on: 
• 
data handling conventions 
• 
outlier handling 
• 
definitions of protocol deviations and PP inclusion 
• 
derivation rules for composite or derived variables 
This SAP will be finalized prior to database lock and prior to performing the final statistical 
analyses, and will be archived with the study documentation. 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 38 of 46 
12. DOCUMENTATION 
12.1 Quality Control and Quality Assurance 
Sponsor through the project management organization has a plan in place for the quality of the 
research being conducted, which describes: 1) How data will be evaluated for compliance with the 
protocol and for accuracy in relation to source documents; 2) The documents to be reviewed (e.g. 
CRFs, source documents, product accountability, final study report), who is responsible, and 3) 
The frequency for reviews will be identified, either in a formal quality management plan or in-
house SOPs. Methods of staff training will be specified. The final report will contain a statement 
for quality assurance (QA) duly signed by the head or designated person of the quality assurance 
department.  
12.2 Data Confidentiality and Audits 
Participant confidentiality along with the information disclosed/provided/produced during study 
will be deidentified before submitting to sponsor and other regulatory authorities. 
The investigator will permit study-related monitoring which is planned, audits, and inspections by 
the IRB, sponsor or its designated study personnel, licensing authority, and QA groups of all study-
related documents (e.g. source documents, regulatory documents, data collection instruments, 
study data, etc.).  
13. ETHICAL CONSIDERATIONS 
13.1 Ethical Conduction and Regulatory Compliance 
The present study will be conducted in accordance with the Declaration of Helsinki (Ethical 
Principles for Medical Research Involving Human Participants, 64th  World Medical Association 
– General Assembly, Fortaleza, Brazil, October 2013), ICH-GCP, New Drugs and Clinical Trials 
Rules, 14 October 2022, Ministry of Health and Family Welfare, Government of India, Indian 
Council of Medical Research (Ethical Guidelines for Biomedical Research on Human Participants, 
2017) and other applicable regulatory requirements. 
13.2 Ethics Committee Approval 
The study protocol and related study documents [including English and vernacular informed 
consent forms (ICFs), translation and back-translation certificates, and drug-related literature] will 
be submitted to, and approved by an IRB in writing. The study will commence only after obtaining 
the written approval of the study protocol and its related documents by the IRB, with or without 
modifications.  

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 39 of 46 
13.3 Clinical Trial Registry 
After receiving at least one ethics committee approval for the study protocol then the proposed 
study will be registered with the clinical trial registry of India (www.ctri.nic.in) before enrolment 
of the first study participant. 
13.4 Protocol Amendments 
All protocol amendments, interfering with the participant’s health interests and involving changes 
in the design of the study or its scientific significance will be implemented only after written 
approval of the sponsor and IRB. All such changes will be documented in the amended version of 
the protocol and a list of changes with reference to the previous version will be generated. Protocol 
amendments only for logistical or administrative changes may be implemented immediately, and 
the amendments will be informed/ notified to IRB. 
13.5 Informed Consent Process 
At screening, all the participants participating in this study will receive study information, verbal 
and written, in their vernacular/understandable language about the purpose and nature of the study 
and its procedures as well as potential risks and benefits associated with study drugs as per the 
participant information sheet (PIS), prior to participation in the study. Participants will be provided 
enough time and opportunity to read the PIS-ICF. Participants will be encouraged to ask questions 
and clarify their doubts before signing or agreeing for signing the PIS-ICF in the presence of an 
investigator or qualified medical personnel.  
The legally acceptable representative (LAR) must sign the informed consent form wherever 
applicable.  
Written informed consent will be obtained from each participant according to good clinical 
practice ICH-GCP, the ethical principles that have their origin in the Declaration of Helsinki, and 
the regulatory and legal requirements of India. 
A copy of the signed and dated written PIS-ICF will be provided to the participant. 
13.6 Participant Participation Compensation 
▪ In case of an injury occurring to the participant during the clinical trial, free medical 
management shall be given as long as required or till such time it is established that the injury 
is not related to the clinical trial, whichever is earlier.  
▪ The participants will be covered under clinical trial liability insurance for medical 
management. 
▪ In the event of a trial-related injury or death, the sponsor or its representatives or the 
investigator or center, as the case may be, in accordance with the “New Drugs and Clinical 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 40 of 46 
Trials Rules, 2022, Department of Health and Family Welfare” issued by the central licensing 
authority, shall provide financial compensation for the injury or death. 
 
13.7 Compensation to The Participants in Clinical Trial Injury 
▪ Compensation for clinical trial–related injury or death will be provided in accordance with the 
New Drugs and Clinical Trials (NDCT) Rules, 2019. If a participant suffers any physical 
injury, permanent disability, or death that is determined to be causally related to the study 
intervention, procedures, or protocol violations, the Sponsor will provide free medical 
management and financial compensation as per the prescribed regulatory formula and 
timelines. The decision regarding causality, eligibility, and compensation will be reviewed by 
the Ethics Committee and the Sponsor, and reported to the Licensing Authority as required 
under the NDCT framework. 
13.8 Insurance Policy and Finance 
The study will be covered by an appropriate insurance contract. Provision will be made for 
insurance to cover the costs of treatment in the event of trial-related injuries in accordance with 
the Indian regulatory requirements.  
The sponsor should provide all study-related finance as per the approved budget. 
14. RECORDING OF DATA, REPORT PREPARATION, AND ARCHIVAL 
14.1  Record Keeping 
The preparation of the clinical trial protocol is in compliance with ICH-GCP, SPIRIT guidelines. 
All other EC dossier documents such as ICD, CRF, PIS, IPIB, study logs, and participant files will 
be prepared as per ICH-GCP guidelines. Data collected on CRFs during the study will be 
documented and identified by the participant number, the participant’s initials, and the study code. 
The investigator will abide to keep the identity by the participant confidentiality agreement, except 
if this information is to be disclosed to assess the safety of the participant or for regulatory 
purposes. The completed CRFs will be checked by the investigator or designee for completeness, 
accuracy, and legibility, for conformance to the source documents. 
14.2 Final Study Report 
The final study report will be prepared as per the ICH-GCP guidelines and will be submitted in 
electronic format. 
 
14.3 Quality Assurance 
All the data related to the study will be checked by the QA before archiving.  
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 41 of 46 
14.4 Record Retention and Archiving 
All the raw and final data generated in connection with this study will be retained in accordance 
with national legislation or as per the sponsor's requirement, whichever is later. All study-related 
documents will be archived, which include the following either at sites and/or sponsor as 
applicable): 
 
▪ IRB approval for the study protocol and its amendments, if any; 
▪ CRFs; 
▪ Signed Informed Consent forms (deidentified); 
▪ Study Protocol and Study Report; 
▪ Study Master File; 
▪ Any other relevant study documents. 
15. TERMINATION OF THE STUDY 
The investigator, sponsor or IRB reserve the right to terminate the study at any time on a safety 
issue. If the investigator terminates or suspends the study without the prior agreement of the 
sponsor, the investigator will promptly inform the sponsor and IRB and provide a detailed written 
explanation of the termination or suspension.  
If the sponsor prematurely terminates or suspends the study, or one or more investigative sites 
participating in the study, the investigator must promptly inform the IRB. The justification for the 
termination will be reported to the IRB within 24 hours and to the central licensing authority within 
30 working days of such termination. The reason for study termination will also be provided to the 
participants. A summary report of the terminated study will be submitted to the central licensing 
authority within 3 months. 
16. PUBLICATION 
The sponsor will hold the right to publish the results of the present study at any time. Any 
information published will not reveal the identity of the participant and their confidentiality will 
be maintained. Investigators may publish or present the results of this study, in either written or 
oral format, only after obtaining the written permission of the sponsor. 
17. IPR VALUE 
The sponsor will hold the right to share IPR rights (if applicable) as per their policy. 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 42 of 46 
18. REFERENCES 
1. United Nations Department of Economic and Social Affairs. World Population Ageing 2019. 
New York: United Nations; 2020. (No DOI—UN reports do not assign DOI) 
2. López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 
2013;153(6):1194–1217. doi: 10.1016/j.cell.2013.05.039 
3. Short KR, Bigelow ML, Kahl J, Singh R, Coenen-Schimke J, Raghavakaimal S, Nair KS. 
Decline in skeletal muscle mitochondrial function with aging in humans. Proc Natl Acad Sci 
U S A. 2005;102(15):5618–23. doi: 10.1073/pnas.0501559102 
4. Cruz-Jentoft AJ, Bahat G, Bauer J, Boirie Y, Bruyère O, Cederholm T, et al. Sarcopenia: 
revised European consensus on definition and diagnosis. Age Ageing. 2019;48(1):16–31. 
doi: 10.1093/ageing/afy169 
5. McCarthy C, Tinsley GM, Bosy-Westphal A, Müller MJ, Shepherd J, et al. Total and regional 
appendicular skeletal muscle mass prediction from dual-energy X-ray absorptiometry body 
composition models. Sci Rep. 2023;13(1):2590. doi: 10.1038/s41598-023-29827-y. 
6. Larsson L, Degens H, Li M, Salviati L, Lee Y, Thompson W, et al. Sarcopenia: Aging-related 
loss 
of 
muscle 
mass 
and 
function. 
Physiol 
Rev. 
2019;99(1):427–511. 
doi: 10.1152/physrev.00061.2017 
7. Studenski S, Perera S, Patel K, Rosano C, Faulkner K, Inzitari M, et al. Gait speed and survival 
in 
older 
adults. 
J 
Gerontol 
A 
Biol 
Sci 
Med 
Sci. 
2011;66(1):203–9. 
doi: 10.1093/gerona/glq163 
8. Harada CN, Natelson Love MC, Triebel KL. Normal cognitive aging. Dialogues Clin 
Neurosci. 2013;15(1):11–27. doi: 10.31887/DCNS.2013.15.1/charada (article DOI resolved 
by publisher) 
9. Fulop T, Larbi A, Dupuis G, Le Page A, Frost EH, Cohen AA, et al. Immunosenescence and 
inflamm-aging. Aging Clin Exp Res. 2017;29(2):215–28.doi: 10.1007/s40520-016-0701-9 
10. Calder PC. Nutrition, immunity and COVID-19. BMJ Nutr Prev Health. 2020;3(1):74–92. 
doi: 10.1136/bmjnph-2020-000085 
11. Kennedy BK, Berger SL, Brunet A, Campisi J, Cuervo AM, Epel ES, et al. Geroscience: 
Linking aging to chronic disease. Cell. 2014;159(4):709–13. doi: 10.1016/j.cell.2014.10.039. 
12. Tipton KD, Wolfe RR. Protein and amino acids for athletes. J Sports Sci. 2004;22(1):65–79. 
doi:10.1080/0264041031000140554 
13. Phillips SM. Dietary protein for athletes. Can J Appl Physiol. 2004;29(3):282–90. 
doi:10.1139/h04-019 
14. Candow DG et al. Creatine supplementation in older adults. Aging Clin Exp Res. 
2014;26(4):475–82. doi:10.1007/s40520-014-0210-4 
15. Kreider RB et al. ISSN position: Creatine safety & efficacy. J Int Soc Sports Nutr. 2017;14:18. 
doi:10.1186/s12970-017-0173-z 
16. Zdzieblik D et al. Collagen peptides improve muscle mass. Br J Nutr. 2015;114(8):1237–45. 
doi:10.1017/S0007114515002810 
17. Shaw G et al. Collagen supports connective tissue recovery. Nutrients. 2017;9(2):67. 
doi:10.3390/nu9020067 
18. Zhu JS et al. Cordyceps pharmacology. J Altern Complement Med. 1998;4(3):289–303. 
doi:10.1089/acm.1998.4.289 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 43 of 46 
19. Chen S et al. Cordyceps immunostimulation. Phytother Res. 2010;24(7):1024–31. 
doi:10.1002/ptr.3060 
20. Holick 
MF. 
Vitamin 
D 
physiology. 
N 
Engl 
J 
Med. 
2007;357:266–81. 
doi:10.1056/NEJMra070553 
21. O’Leary F, Samman S. Vitamin B12 and cognition. Nutrients. 2010;2(3):299–316. 
doi:10.3390/nu2030299 
22. Montgomery SA et al. ALCAR in cognitive decline. Int Clin Psychopharmacol. 2003;18(1):9–
14. doi:10.1097/00004850-200301000-00002 
23. Malaguarnera M. Carnitine neuroprotection. Curr Pharm Des. 2012;18(27):4021–6. 
doi:10.2174/138161212802002608 
24. Glade MJ, Smith K. Phosphatidylserine cognition. Nutr Neurosci. 2015;18(2):79–90. 
doi:10.1179/1476830514Y.0000000116 
25. Jäger R et al. Phosphatidylserine supplementation. J Int Soc Sports Nutr. 2007;4:5. 
doi:10.1186/1550-2783-4-5 
26. Sengupta K et al. Boswellia clinical trial. Int J Med Sci. 2008;5(6):366–72. 
doi:10.7150/ijms.5.366 
27. Abdel-Tawab M. Boswellia pharmacology review. Planta Med. 2020;86(3):224–40. 
doi:10.1055/a-1062-4050 
28. Stough C et al. Bacopa cognitive trial. Psychopharmacology. 2001;156(4):481–4. 
doi:10.1007/s002130100815 
29. Pase MP et al. Bacopa systematic review. J Altern Complement Med. 2012;18(7):647–66. 
doi:10.1089/acm.2011.0367 
30. Tan MS et al. Ginkgo meta-analysis. J Alzheimers Dis. 2015;43:589–603. doi:10.3233/JAD-
141446 
31. Maclennan KM et al. Bioactivity of Ginkgo. Biochim Biophys Acta. 2002;1582(1–3):165–
72. doi:10.1016/S0167-4889(02)00245-0 
32. Choudhary D et al. Ashwagandha stress/cognition. J Clin Psychopharmacol. 2017;37(5):520–
5. doi:10.1097/JCP.0000000000000730 
33. Wankhede S et al. Ashwagandha strength trial. J Int Soc Sports Nutr. 2015;12:43. 
doi:10.1186/s12970-015-0104-9 
34. Fitton 
JH. 
Fucoidan 
biological 
activity. 
Mar 
Drugs. 
2011;9(10):1731–60. 
doi:10.3390/md9101731 
35. Li B et al. Seaweed polysaccharides. J Appl Phycol. 2008;20:449–54. doi:10.1007/s10811-
007-9272-5 
36. Hewlings 
SJ, 
Kalman 
DS. 
Curcumin 
review. 
Foods. 
2017;6(10):92. 
doi:10.3390/foods6100092 
37. Aggarwal BB et al. Curcumin molecular targets. Biochem Pharmacol. 2009;76:1590–612. 
doi:10.1016/j.bcp.2008.08.031 
38. Masibo M, He Q. Mango bioactives: Mangiferin & antioxidant capacity. Food Res Int. 
2008;41(10):928–35. doi:10.1016/j.foodres.2008.04.006 
39. Guha S et al. Mangiferin reduces oxidative biomarkers. J Agric Food Chem. 
2010;58(7):4104–12. doi:10.1021/jf903138p 
40. Gröber U, Schmidt J, Kisters K. Magnesium in human health. Nutrients. 2015;7(9):8199–226. 
doi:10.3390/nu7095388 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 44 of 46 
41. Volpe 
SL. 
Magnesium 
& 
performance. 
Magnes 
Res. 
2015;28(4):127–32. 
doi:10.1684/mrh.2015.0407 
42. Baur JA et al. Resveratrol improves metabolic performance. Nature. 2006;444:337–42. 
doi:10.1038/nature05354 
43. Smoliga JM et al. Human safety & dose of resveratrol. Mol Nutr Food Res. 2011;55(8):1129–
41. doi:10.1002/mnfr.201100143 
44. Li Y et al. Quercetin review. Nutrients. 2016;8(3):167. doi:10.3390/nu8030167 
45. Boots AW et al. Molecular mechanisms of quercetin. Eur J Pharmacol. 2008;585:325–37. 
doi:10.1016/j.ejphar.2008.03.008 
46. Candow DG, Chilibeck PD, Forbes SC. Creatine supplementation and aging muscle. Aging 
Clin Exp Res. 2014;26(4):475–82. doi:10.1007/s40520-014-0210-4 
47. Devries MC, Phillips SM. Creatine supplementation and resistance exercise in older adults. J 
Cachexia Sarcopenia Muscle. 2014;5(2):97–110. doi:10.1007/s13539-014-0133-5 
48. Morton RW, McGlory C, Phillips SM. Nutritional protein and skeletal muscle anabolism. 
Nutrients. 2015;7(3):1859–82. doi:10.3390/nu7031859 
49. Chilibeck PD, Kaviani M, Candow DG, Zello GA. Incorporating protein with creatine: aging 
and performance outcomes. Nutrients. 2017;9(11):1307. doi:10.3390/nu9111307 
50. Pase MP, Kean J, Sarris J, et al. Bacopa monnieri and cognitive performance: systematic 
review. J Altern Complement Med. 2012;18(7):647–66. doi:10.1089/acm.2011.0367 
51. Tan MS, Yu JT, Tan CC, et al. Efficacy of Ginkgo biloba in neurocognition: meta-analysis. J 
Alzheimers Dis. 2015;43(2):589–603. doi:10.3233/JAD-141446 
52. Montgomery SA, Thal LJ, Amrein R. Acetyl-L-Carnitine in elderly cognitive disorders: 
review. Int Clin Psychopharmacol. 2003;18(1):9–14. doi:10.1097/00004850-200301000-
00002 
53. Chen S, Li Z, Krohn J, et al. Cordyceps immunomodulation. Phytother Res. 2010;24(7):1024–
31. doi:10.1002/ptr.3060 
54. Choudhary D, Bhatt K, Pandey A. Withania somnifera in stress and immunity. J Clin 
Psychopharmacol. 2017;37(5):520–5. doi:10.1097/JCP.0000000000000730 
55. Li Y, Yao J, Han C, et al. Quercetin: bioactivities and anti-inflammatory potential. Nutrients. 
2016;8(3):167. doi:10.3390/nu8030167 
56. Smoliga JM, Baur JA, Hausenblas HA. Resveratrol human evidence and dosing. Mol Nutr 
Food Res. 2011;55(8):1129–41. doi:10.1002/mnfr.201100143 
57. Kreider RB, Kalman DS, Antonio J, et al. International Society of Sports Nutrition position 
stand: Safety and efficacy of creatine supplementation. J Int Soc Sports Nutr. 2017;14:18. 
doi:10.1186/s12970-017-0173-z 
58. Choudhary D, Bhatt K, Pandey A. Efficacy and safety of Ashwagandha root extract in 
reducing stress and anxiety in adults: A double-blind randomized controlled trial. J Clin 
Psychopharmacol. 2017;37(5):520–5. doi:10.1097/JCP.0000000000000730 
 
APPENDICES 
Appendix-A: Investigator Approval for Protocol 
Appendix-B: Schedule of Study Events 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 45 of 46 
APPENDIX-A: INVESTIGATOR APPROVAL FOR PROTOCOL 
 
I, the undersigned, have read and understood this protocol, and hereby agree to conduct the 
study in accordance with current protocol, Declaration of Helsinki (Ethical Principles for 
Medical Research Involving Human Participants, 64th World Medical Association – General 
Assembly, Fortaleza, Brazil, October 2013), Integrated Addendum to ICH E6 (R1): Guideline 
for Good Clinical Practice, E6(R2), Current Step 4 version dated 9 November 2016, New 
Drugs and Clinical trials Rules, October 2022, Central Licensing Authority, Ministry of Health 
and Family Welfare, Government of India, Indian Council of Medical Research (Ethical 
Guidelines for Biomedical Research on Human Participants, 2017) and applicable regulatory 
Guidelines.        
I agree to comply with all relevant standard operating procedures required for the conduct of 
this study and to ensure that all associates participating in the conduct of this study are 
informed regarding their obligations. 
I understand that the information in this protocol is confidential and should not be disclosed, 
other than to those directly involved in the execution or the ethical review of the trial, without 
written authorization from GP Life Healthcare Pvt. Ltd. 
I understand that the trial can get suspended or prematurely terminated due to valid reasons 
and will be communicated to all stakeholders. I understand my responsibility to communicate 
immediately in written, if in case my participation in the trial needs to be discontinued. 
 
 
 
___________________________                        
Name of the Principal Investigator 
 
 
 
          
               
 
_________________ 
Signature and date 
 
 
 
 
 

 
A proof-of-concept clinical study to evaluate the impact of a combination 
nutraceutical on muscle function, cognitive performance, and immune 
resilience in older adults. 
PROTOCOL 
Version: 1.00  
dated 19th November 2025 
Study Code: 
GP/CT/25-26/001 
Sponsor: GPLIFE HEALTHCARE 
PRIVATE LIMITED. 
 
CONFIDENTIAL                                                                                                         
Page 46 of 46 
APPENDIX-B:  SCHEDULE OF STUDY EVENTS 
 
Table 6: Schedule of study events 
 
                                       Day     
Parameter 
Screening Day 1 Day 30 Day 60 
Informed Consent 
√ 
 
 
 
Demographics 
√ 
 
 
 
Medical History / Previous Medication 
√ 
 
 
 
Concomitant Diseases/ Medication  
√ 
 
 
 
Inclusion /Exclusion Criteria Review 
√ 
 
 
 
Randomization 
 
√ 
 
 
Dispensing of IP 
 
√ 
√ 
 
Clinical examination   
√ 
√ 
√ 
√ 
Physical Examination   
√ 
√ 
√ 
√ 
Vitals 
√ 
√ 
√ 
√ 
Assessment of adverse events 
 
√ 
√ 
√ 
Assessment of rescue medications 
 
√ 
√ 
√ 
Tolerability 
 
 
√ 
√ 
Compliance of IP 
 
 
√ 
√ 
Assessment of 6-minute walk distance (6MWD) 
(meters) 
√ 
 
 
√ 
 Assessment of 1RM (kg) with the leg press 
√ 
 
 
√ 
Assessment of Muscle volume 
√ 
 
 
√ 
Assessment of Fluid Composite score and assessment 
of executive function, attention and processing speed, 
working memory 
√ 
 
 
√ 
Assessment of plasma p-tau181 
√ 
 
 
√ 
Assessment of CD4/CD8 ratio, CD45, CD3, CD4, 
CD8, T cells, B cells, NK cells, 
lymphocyte/neutrophil ratio 
√ 
 
 
√ 
Assessment of serum hs-CRP, TNF-alpha, IL-6 
√ 
 
 
√ 
Assessment of plasma MDA and SOD, NAD+ from 
whole blood 
√ 
 
 
√ 
Assessment of Body Weight (kg), BMI (Kg/m2) and 
hip, waist, chest circumferences (cm) 
√ 
 
 
√ 
Assessment of complete blood count (CBC), Renal 
Function Test (RFT). Liver Function Test (LFT), 
Thyroid profile, lipid profile, electrolytes. 
√ 
 
 
√