Clinical trial NCT05429073
Conditions: Healthy Volunteers
Keywords: RGLS8429 Autosomal Dominant Polycystic Kidney Disease ADPKD
Sponsor: Regulus Therapeutics Inc.
Start date: 2022-06-10; Completion date: 2022-09-08
Enrollment: 32 participants; Allocation: RANDOMIZED; Intervention model: SEQUENTIAL

Brief summary:
Primary Objective

• To assess the safety and tolerability of single ascending doses of RGLS8429

Secondary Objectives

* To identify dose-limiting toxicity (DLT) and to determine the maximum tolerated dose (MTD) of a single SC dose of RGLS8429
* To characterize the pharmacokinetic (PK) properties of RGLS8429

Detailed description:
In this randomized, double-blind, placebo-controlled Phase 1 study, a single ascending dose of RGLS8429 or placebo will be administered via subcutaneous (SC) injection to healthy volunteers to evaluate the safety, tolerability, and PK of RGLS8429.

Arms: arm 1: Eligible participants will receive subcutaneous injection of the first dose level RGLS8429 or placebo arm 2: Eligible participants will receive subcutaneous injection of the second dose level RGLS8429 or placebo arm 3: Eligible participants will receive subcutaneous injection of the third dose level RGLS8429 or placebo arm 4: Eligible participants will receive subcutaneous injection of the fourth dose level RGLS8429 or placebo
Interventions: intervention 1: RGLS8429 intervention 2: Placebo volume-matching RGLS8429 dose
Intervention descriptions: intervention 1: solution for subcutaneous injection intervention 2: solution for subcutaneous injection

Protocol documents:
Prot_000.pdf: 
RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
1 
6 June 2022 
TITLE PAGE 
A Phase 1, Double-Blind, Placebo-Controlled, Single 
Ascending Dose Study in Healthy Volunteers to Evaluate the 
Safety, Tolerability, and Pharmacokinetics of RGLS8429 
 
 
 
 
 
INVESTIGATIONAL PRODUCT (IP) 
 
RGLS8429 
PROTOCOL NUMBER: 
 
RGLS8429-01 v3.0 
PROTOCOL DATE: 
 
6 June 2022 
IND NUMBER: 
 
158708 
SPONSOR NAME / ADDRESS: 
 
 
 
Regulus Therapeutics Inc. 
4224 Campus Point Court 
Suite 210 
San Diego, CA 92121 
United States 
 
 
 
 
CONFIDENTIAL 
This protocol is provided to you as an Investigator, potential Investigator, or consultant for 
review by you, your staff, and ethics committee/institutional review board. The information 
contained in this document is regarded as confidential and, except to the extent necessary 
to obtain informed consent, may not be disclosed to another party unless such disclosure is 
required by law or regulations. Persons to whom the information is disclosed must be 
informed that the information is confidential and may not be further disclosed by them. 
 
 


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
2 
6 June 2022 
DOCUMENT HISTORY 
Version 
Version Date 
Summary of Changes and Rationale 
Version 1.0 
9 March 2022 
Not Applicable 
Version 2.0 
09 May 2022 
The protocol is being amended to: 
1. NCI CTCAE Version 5.0 scale to 
rate adverse event severity in this 
healthy volunteer study, is replaced 
with the Guidance for Industry: 
Toxicity Grading Scale for Healthy 
Adult and Adolescent Volunteers 
Enrolled in Preventive Vaccine 
Clinical Trials (September 2007) for 
all the parameters except for serum 
creatinine.  The KDIGO Clinical 
Practice Guideline for Acute Kidney 
Injury (https://kdigo.org/wp-
content/uploads/2016/10/KDIGO-
2012-AKI-Guideline-English.pdf) 
will be used to rate serum creatinine 
adverse event severity. Add AKI 
criteria to Section 3.2, Dose 
Escalation and Stopping Rules.  
2. Increase the sampling window for 
the collection of post-dose 
pharmacokinetic plasma samples 
when multiple procedures are being 
performed at the same nominal 
timepoints.   
3. Clarify that the NuvaRing is an 
acceptable depot contraceptive. 
4. Allow the pre-dose ECG tracing to 
be obtained and the SARA test to be 
performed, prior to the start of 
Holter monitoring at the 60 minute 
pre-dose timepoint on Day 1. 
Version 3.0 
6 June 2022 
The protocol is being amended to: 
1. Change the RGLS8429 dose for 
FRKRUWIURP 5.0 mg/kg to 
4.0 mg/kg. The dose of 4mg/kg has 
been selected for cohort 4 and is 
within the dose range supported by 
non-clinical toxicology and 


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
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6 June 2022 
Version 
Version Date 
Summary of Changes and Rationale 
pharmacokinetic studies, where 
RGLS8429 demonstrated dose-
proportional exposure, and low 
variability. Hence, an interim PK 
analysis is not needed for dose level 
selection for cohort 4.  
2. Correct minor typographical errors 
and clarify the description of the 
collection of potential renal 
biomarkers in urine 
 


RGLS8429  
Protocol RGLS8429-01 v3.0 
Regulus Therapeutics Inc.
Confidential and Proprietary 
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SPONSOR SIGNATURE PAGE 
 
 
 
 
 
Signature of Sponsor Lead Physician 
 
DATE 
 
Rekha Garg, MD, MS  
Printed Name of Sponsor Lead Physician 
By my signature, I indicate I have reviewed this protocol and find its content to be acceptable.  
 
 
 
 
 
Signature of Sponsor VP Clinical Development 
 
DATE 
 
Karl Cremer, PharmD  
Printed Name of Sponsor VP Clinical Development 
By my signature, I indicate I have reviewed this protocol and find its content to be acceptable.  




RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
5 
6 June 2022 
SITE PRINCIPAL INVESTIGATOR SIGNATURE PAGE 
 
 
 
 
Signature of Site Principal Investigator 
 
DATE 
 
 
Printed Name of Site Principal Investigator 
Institution Name: _______________________________________ 
By my signature, I agree to personally supervise the conduct of this study at my study site and to 
ensure its conduct is in compliance with the protocol, informed consent, Institutional Review Board 
(IRB)/Ethics Committee (EC) procedures, instructions from Regulus representatives, the 
Declaration of Helsinki, International Conference on Harmonisation (ICH) Good Clinical Practices 
Guidelines, and local regulations governing the conduct of clinical studies. 
 


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
6 
6 June 2022 
PROTOCOL SYNOPSIS 
Study Title:  
A Phase 1, Double-Blind, Placebo-Controlled, Single Ascending Dose Study in Healthy 
Volunteers to Evaluate the Safety, Tolerability, and Pharmacokinetics of RGLS8429 
Protocol Number: RGLS8429-01 (IND 158708) 
Indication: Treatment of Autosomal Dominant Polycystic Kidney Disease (ADPKD) 
Study Type: Phase 1, dose range-ranging study in healthy volunteers 
Study Population: Approximately 32 healthy volunteers 
Study Sites: Phase 1 Clinical Research Unit in the USA 
Sponsor: Regulus Therapeutics Inc. 
Study Drugs:  
RGLS8429 injection will be provided in 2 mL clear glass vials containing sufficient volume to 
extract the labeled volume of 1 mL of 150mg/mL of RGLS8429 in 0.3% saline.  
Placebo injection will be provided in 2 mL clear glass vials containing sufficient volume to extract 
the labeled volume of 1 mL of 1.5 μg/mL of riboflavin in 0.9% sodium chloride. 
Both RGLS8429 and placebo solutions are clear and colorless to pale yellow. 
Study Rationale:  
ADPKD is a monogenic systemic disease characterized by slowly progressive, bilateral 
enlargement of fluid-filled polycystic kidneys where increase in total kidney volume occurs long 
before detectable decline of renal function. Fifty percent of ADPKD patients develop end-stage 
renal disease (ESRD) by the age of 60, accounting for 5% of all ESRD patients in the United States 
(US) requiring renal replacement therapy such as dialysis or transplant.  
RGLS8429 is a potent and selective anti-miR-17 oligonucleotide that dose-dependently inhibits the 
function of miR-17 in kidney cells in vitro and in animal models of polycystic kidney disease 
(PKD) in vivo. The expression of the miR-17 family of microRNAs has been shown to be 
increased in kidney samples from both mouse ADPKD models and patients with ADPKD. Kidney-
specific knockdown of the miR-17~92 cluster attenuates disease progression and increases overall 
survival in multiple transgenic mouse models of PKD and subcutaneous (SC) dosing with 
RGLS8429 resulted in improvement in two different mouse models of PKD. In the Pkd1F/RC mouse 
model, RGLS8429 significantly reduced kidney weight-to-body weight ratio, serum blood urea 
nitrogen (BUN), kidney cyst index, and kidney cyst proliferation. In the Pcy/DBA mouse model, 
RGLS8429 significantly reduced kidney weight-to-body weight ratio, Taken together, these studies 
demonstrate that miR-17 is a promising therapeutic target for the treatment of ADPKD. 
Study Objectives: 
Primary Objective 
x 
To assess the safety and tolerability of single ascending doses of RGLS8429  


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
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Secondary Objectives 
x 
To identify dose-limiting toxicity (DLT) and to determine the maximum tolerated dose 
(MTD) of a single SC dose of RGLS8429 
x 
To characterize the pharmacokinetic (PK) properties of RGLS8429 
Exploratory Objective 
x 
To characterize multiple potential renal biomarkers  
Study Endpoints: 
Primary Endpoint 
x 
Incidence of adverse events (AEs) and change in safety laboratory test results, vital signs, 
Scale for the Assessment and Rating of Ataxia (SARA), and electrocardiogram (ECG) 
parameters and Holter monitoring over time  
Secondary Endpoints 
x 
Incidence of DLT and determination of MTD 
x 
PK parameters of RGLS8429 including: 
o Maximum observed concentration (Cmax)  
o Time to maximum observed concentration (Tmax)  
o Area under the concentration–time curve up to 24 hr post dose (AUC0-24) 
o Area under the concentration–time curve up to the last quantifiable concentration 
(AUC0-t) 
o Area under the concentration–time curve extrapolated to infinity (AUCinf)  
o Half-life (t½)  
o Apparent clearance (CL/F)  
o Volume of distribution (Vz/F) 
o Fraction of unchanged RGLS8429 excreted in the urine (fe) 
o Total amount of unchanged RGLS8429 excreted in the urine (Ae) 
Exploratory Endpoints 
x 
Characterize multiple potential renal biomarkers in urine (e.g., polycystin 1 [PC1], 
polycystin 2 [PC2], neutrophil gelatinase-associated lipocalin [NGAL], kidney injury 
molecule-1 [KIM-1], monocyte chemoattractant protein-1 [MCP-1], beta-2 microglobulin 
[B2M], and clusterin [CLU]) and in serum (e.g., Cystatin-C, Insulin-like Growth Factor 
Binding Protein Acid Labile Subunit [IGFALS], albumin [Alb], copeptin [CT-proAVP], 
N-acetyl-1-methylhistidine, and others) 
Study Design and Methodology: 
In this randomized, double-blind, placebo-controlled Phase 1 study, a single ascending dose of 
RGLS8429 or placebo will be administered via SC injection to healthy volunteers to evaluate the 
safety, tolerability, and PK of RGLS8429.  


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
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6 June 2022 
Eight subjects will be randomized 3:1 RGLS8429:Placebo into each of 4 sequential cohorts (32 
subjects in total).  
Cohort 1 
0.5 mg/kg RGLS8429 or placebo 
Cohort 2 
1.0 mg/kg RGLS8429 or placebo 
Cohort 3 
2.0 mg/kg RGLS8429 or placebo 
Cohort 4 
4.0 mg/kg RGLS8429 or placebo 
In each cohort, 6 subjects will be randomized to receive RGLS8429 and 2 will be randomized to 
receive placebo. Enrollment into subsequent cohorts will be initiated after review of safety (AEs 
and safety laboratory tests) from the prior cohort. 
For Cohort 1, the first two subjects will receive RGLS8429 or placebo (randomized such that one 
subject receives RGLS8429 and one receives placebo) and this sentinel group will be monitored for 
48 hr prior to dosing the remaining subjects in the cohort. 
An unblinded pharmacist will be responsible for preparing RGLS8429 or placebo and providing 
the study drug to blinded study personnel for administration according to the randomization 
procedures.  
Subjects will be admitted to the study center on Day -1, one day prior to the day of study drug 
administration (Day 1), and will be discharged on Day 2, no less than 6 hr after the collection of 
the 24-hr PK sample. Subjects will be followed for a total of 4 weeks after dosing and will return to 
the site for follow-up visits weekly from Weeks 1 through 4. The sentinel subjects will remain in 
the clinic for 48 hr after dosing. 
Dose Escalation and Stopping Rules: 
Prior to dose escalation, the Sponsor and the Investigator will perform a blinded review of the 
safety data (e.g., AE and safety laboratory tests through Day 15) for all subjects at the current dose 
level to make a dose escalation decision.  
If 2 or PRUHVXEMHFWVDWWKHVDPHGRVHOHYHOH[SHULHQFHWKHVDPHRUVLPLODU*UDGH$(V
LQFOXGLQJFOLQLFDOO\VLJQLILFDQWODERUDWRU\$(VRULIVXEMHFWH[SHULHQFHVD*UDGH$(RUD
serious adverse event (SAE) judged to be related to study drug, the Sponsor’s Lead Physician will 
be unblinded to the treatment. Also, if 2 subjects develop stage 1 AKI or 1 subject develops stage 2 
AKI the Sponsor’s Lead Physician will be unblinded to the treatment. 
The Sponsor’s Lead Physician will determine whether the nature, severity, or number of event(s) 
warrants: 
1. Continuing with the study as planned 
2. Continuing with the study, but add additional safety evaluation(s) 
3. Suspend dose escalation and enroll additional subjects at the same dose level 
4. Add an intermediate-dose cohort or adjust the next dose level 
5. Suspend or terminate the study 
Study Population:  
Approximately 32 healthy volunteers 


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
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6 June 2022 
Inclusion Criteria  
1. Male or female, 18 to 55 years of age  
2. Continuous nonsmoker who has not used nicotine-containing products for at least 3 months 
prior to dosing 
3. Body mass index (BMI) 18 to 35 kg/m2 
4. Medically healthy, with no clinically significant medical history in the opinion of the 
Investigator 
5. Estimated glomerular filtration rate (eGFR)  90 mL/min/1.73 m2 (see Section 2.3.3) 
6. Platelet count within the normal limits 
7. Hemoglobin concentration within the normal limits 
8. Liver function tests (alanine aminotransferase [ALT], aspartate aminotransferase [AST], 
alkaline phosSKDWDVH>$/3@DQGELOLUXELQ>WRWDODQGGLUHFW@PXVWEH the upper limit of 
normal (ULN)  
9. Female subjects of childbearing potential must not be lactating and must have no plans to 
become pregnant during the course of the study through 28 days after the administration of 
study drug. Female subjects of childbearing potential who are heterosexual must agree to use 
one of the following methods of contraception considered to be highly effective (i.e., results in 
<1% failure rate when used consistently and correctly) from Screening through 28 days after 
the administration of study drug 
a. Intrauterine device (IUD) or intrauterine system (IUS) in place for at least 3 months prior 
to study drug administration 
b. Partner has had a vasectomy. Vasectomy in the partner is considered to be highly effective 
only if the partner is the sole sexual partner of the female subject of childbearing potential 
and had his vasectomy performed 6 months or more prior to randomization. 
c. Stable hormonal contraception associated with inhibition of ovulation (with approved oral, 
transdermal, or depot [e.g., NuvaRing, depot injection, etc.] regimen) for at least 3 months 
prior to study drug administration 
10. A female subject of non-childbearing potential must have undergone one of the following 
sterilization procedures at least 6 months prior to the administration of study drug: 
a. Hysterectomy  
b. Bilateral oophorectomy  
c. Bilateral tubal occlusion 
d. Bilateral salpingectomy 
or be postmenopausal with no menstrual periods for at least 1 year prior to the 
administration of study drug. 
11. Non-vasectomized heterosexual male subjects must agree to use a condom with spermicide or 
abstain from sexual intercourse from Day 1 until 28 days after the administration of study drug. 
(No restrictions are required for a vasectomized heterosexual male provided his vasectomy was 
performed 6 months or more prior to study start. A heterosexual male who has been 


RGLS8429  
Protocol RGLS8429-01 v3.0 
 
Regulus Therapeutics Inc. 
Confidential and Proprietary 
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6 June 2022 
vasectomized less than 6 months prior to study start must follow the same restrictions as a non-
vasectomized heterosexual male.) 
12. Male and female subjects must agree not to donate sperm or preserve eggs (ova), respectively, 
from Day 1 until 28 days after the administration of study drug 
13. Must agree not to donate blood in the 28 days prior to randomization or plasma in the 7 days 
prior to randomization through the end of study (EOS) visit 
14. Seated blood pressure (systolic/diastolic) is  90/40 mmHg and  140/90 mmHg 
15. Seated heart rate is  40 bpm and 99 bpm 
16. Fridericia-corrected QT (QTcF) interval is < 460 msec (males) or < 480 msec (females) and 
without any ECG findings deemed clinically significant by the Investigator 
17. Must understand and consent to the study procedures explained in the informed consent form 
(ICF) and be willing and able to comply with the protocol 
Exclusion Criteria 
1. Subject is mentally incapacitated or has significant emotional problems 
2. Any medical condition or social circumstance that, in the opinion of the Investigator, may 
make the subject unlikely to complete the study or comply with study procedures and 
requirements, or may pose a risk to the subject’s safety 
3. History of malignancy, except for successfully treated squamous or basal cell carcinoma skin 
cancer  
4. History or presence of opportunistic infection, in the opinion of the Investigator 
5. History or presence of any clinically significant local infection (e.g., cellulitis, abscess) or 
systemic infection (e.g., septicemia) within 3 months prior to Screening 
6. Fever (body temperature  100.4°F) or symptomatic viral or bacterial infection within 14 days 
prior to Screening, at Screening, at Day -1 check-in, or at Day 1 prior to dosing 
7. History of any clinically significant reaction to previous injection of an oligonucleotide product 
in the opinion of the Investigator 
8. History or presence of alcoholism or drug abuse within the past 2 years prior to Screening 
9. Positive urine drug or alcohol results  
10. +DVSURWHLQRQXULQHGLSVWLFN 
11. Positive results at Screening for human immunodeficiency virus (HIV), hepatitis B surface 
antigen (HBsAg), or hepatitis C virus (HCV) antibody 
12. Positive urine cotinine  
13. Unable to refrain from, or anticipates the use of, any drug, including prescription and 
non-prescription medications and herbal remedies, from 14 days prior to dosing until 
completion of the EOS visit, except as approved by the Investigator  
14. History of any major surgical procedure within 3 months prior to dosing (excluding minor 
cosmetic surgery or minor dental procedures) 


RGLS8429  
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Confidential and Proprietary 
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6 June 2022 
15. Subjects with tattoo(s) or scarring at or near the site of SC injection or any other condition that 
may interfere with injection site examination(s), in the opinion of the Investigator 
16. Participation in another clinical trial and/or exposure to any investigational drug or approved 
therapy for investigational use within 28 days or 5 half-lives of the investigational drug’s 
dosing, whichever is longer, prior to dosing. The 28-day and 5-half-life windows will be 
calculated from the date of the last dosing in the previous study to Day 1 of the current study. 
Statistical Methods: 
Tabular summaries and analysis results will be generated by dose level for RGLS8429 and placebo 
(pooled across all cohorts). Descriptive statistics by dose level and treatment group will be 
tabulated by visit. 
Sample Size  
Formal sample size calculations were not performed for this study. The dose cohort size is 8 
subjects randomized 3:1 to RGLS8429 versus placebo. The size of each dose cohort was chosen to 
provide sufficient information to allow assessment of the safety and tolerability of RGLS8429.  
Safety Analysis  
Safety data (e.g., frequencies of treatment-emergent adverse events [TEAEs] and SAEs, changes 
from baseline in SARA test scores, safety laboratory test results, vital signs, ECG parameters and 
Holter monitoring) will be summarized descriptively by dose level and treatment group, as 
appropriate. No formal statistical tests will be conducted to assess the safety or tolerability of 
RGLS8429.  
Pharmacokinetic Analysis 
PK samples will be analyzed for RGLS8429 concentration using a validated, sensitive, specific 
bioanalytical method. The laboratory analyzing the PK samples will be unblinded, so PK analysis 
is performed only on subjects receiving RGLS8429. Placebo samples will be analyzed if necessary. 
Using non-compartmental methods, the plasma and urine concentration versus time data will be 
used to derive the following PK parameters: Cmax, Tmax, AUC0-24, AUC0-t, AUCinf, t½, CL/F, Vz/F, 
fe, and Ae.  
Dose proportionality will be explored using the power model for Cmax, AUC0-24, AUC0-t, and 
AUCinf s data permit. 
Biomarker Analysis 
Descriptive statistics will be used to characterize urine (e.g., PC1, PC2, NGAL, KIM-1, MCP-1, 
B2M, and CLU) and serum (e.g., Cystatin-C, IGFALS, Alb, CT-proAVP, N-acetyl-1-
methylhistidine, and others) biomarkers in all subjects.  
 


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Protocol RGLS8429-01 v3.0 
 
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Table 1: 
Schedule of Assessments and Procedures 
 
Screen 1 
Treatment 
Follow-Up 
Week 
 
 
 
1 
2 
3 
4/EOS 
Days 
-28 to -2 -1 
1 
2 
8 15 22 
29 
Hour 
 
 
0 2 0.5 
1 
2 
3 
4 
5 
6 
8 
10 12 16 24 
 
 
 
 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical History 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Physical Examination 3 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
X X 
X 
X 
Weight and Height 4 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
X 
Vital Signs 5 
X 
X 
X 
 
X X 
 
X 
 
 
X 
 
 
 
X 
X X 
X 
X 
ECGs 6 
X 
 
X 
 
 
 
 
X 
 
X 
 
 
X 
 
X 
 
 
 
 
Hematology 7 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X X 
X 
X 
Chemistry 8 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X X 
X 
X 
Coagulation 9 
 
X 
 
 
 
 
 
X 
 
 
 
 
 
 
X 
 
 
 
X 
Urinalysis 10 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X X 
X 
X 
Lipids 11 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
X 
Pregnancy Test 12 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
Urine Drug and Alcohol Screen 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
HBsAg, HCV, HIV Screen 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/Exclusion Review 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Randomization 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
SARA Assessment 13 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
X 
Adverse Events 
 
 
X 
X X 
X 
X 
Concomitant Medications 
X 
X 
X 
X X 
X 
X 
Study Drug Administration 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
PK Plasma Collection 14, 19 
 
 
X 
 
 
X 
 
X 
 
X X 
 
X 
 
X 
 
X 
 
 
24-Hr Urine Collection 15, 19 
 
 
X 
 
 
 
 
Serum biomarkers 16, 19 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine biomarkers 17, 19 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Holter monitoring 18 
 
 
X 
 
 
 
 
Admission to Clinic 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Discharge from Clinic 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
Clinic Visit 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X X 
X 
X 


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Abbreviations: ECG, electrocardiogram; EOS, end of study or early termination visit; HBsAg, hepatitis B surface antigen; HCV, hepatitis C virus; HIV, human 
immunodeficiency virus; PK, pharmacokinetics; SARA=Scale for the Assessment and Rating of Ataxia. 
1 
Screening Period within 28 days prior to study drug administration (Baseline = Day -1 or Day 1 when test/procedure performed). 
2 
Ensure that all Day 1 procedures scheduled to be completed prior to dosing are completed prior to dosing. 
3 
Complete physical examination at Screening and Week 4/EOS Visit. Limited physical examination at other visits. The limited physical examination will 
be focused on general appearance, the respiratory, neurological, and cardiovascular systems, and subject-reported symptoms  
4 
Measure weight at all visits indicated. Height at Screening only. Calculate BMI at Screening Visit. 
5 
Systolic (SBP) and diastolic (DBP) blood pressure measurements, heart rate (HR), body temperature, and respiratory rate (RR). 
6 
Obtain a 12-lead ECG during Screening and print and review ECG tracings from the Holter monitor pre-dose and 4 hr, 6 hr, 12 hr, and 24 hr post-dose. 
Record new or worsening clinically significant abnormalities as an AE. 
7 
Full hematology panel including hemoglobin (Hb), hematocrit (Hct), red blood cell (RBC) count, red cell indices, white blood cell (WBC) count including 
absolute cell counts, with differential, and platelet count. 
8 
Fasting chemistry panel including total protein, glucose, uric acid, creatine kinase (CK), lactate dehydrogenase (LDH), sodium, potassium, chloride, 
bicarbonate, calcium, phosphate, BUN, creatinine (Scr), and hepatic function panel (albumin, total bilirubin, direct bilirubin, ALT, AST, ALP, and gamma-
glutamyl transferase [GGT]). 
9 
Prothrombin time (PT), activated partial thromboplastin time (aPTT), international normalized ratio (INR), and fibrinogen will be collected on Day -1; 
Day 1, 4 hr post-dose; and Day 2, 24 hr post-dose; and at Week 4/EOS.  
10 Urinalysis panel including specific gravity, pH, protein, glucose, ketones, blood (Hb), leukocyte esterase nitrite, bilirubin, and urobilinogen. If protein, 
blood, or leukocyte esterase nitrate parameters are abnormal a microscopic examination will be completed.  
11 Fasting lipid panel including total cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL). 
12 Only female subjects of childbearing potential. A serum pregnancy test will be performed at Screening and urine pregnancy tests will be performed at the 
remaining specified visits. 
13 Perform the SARA assessment pre-dose on Day 1 and 4-6 hr post-dose on Day 1; on Day 2, 1 hr after the 24-hr PK sample collection; and at Week 4/EOS 
Visit. 
14 Pre-dose plasma PK sample on Day 1 should be obtained within 60 minutes prior to dosing. Post-dose plasma samples should be obtained within the 
following time margins: collections at 2, 4, 6, and 8 hr ±15 minutes, 12 hr ±30 minutes, and 24 hr ±60 minutes.  
15 24-hr urine collection for PK analysis will begin immediately after study drug dosing on Day 1 (0-24 hr). The subject should void immediately before 
dosing (before the 24-hr urine collection is started). Last void will be at 24 hours post dose.  
16 Serum sample for exploratory biomarkers (e.g., Cystatin-C, IGFALS, Alb, CT-pro-AVP, N-acetyl-1-methylhistidine, and others).  
17 Collect at least 150 mL (more if possible) of mid-stream free-flow urine for analysis of exploratory biomarkers (e.g., PC1, PC2, NGAL, KIM-1, MCP-1, 
B2M, and CLU during Screening and on Day -1. These samples must NOT be the first urination of the day (to avoid too much debris in the sample). 
18 Start Holter monitoring 1 hr before study drug administration on Day 1 and continue for 30 minutes after the 24-hr PK sample is obtained on Day 2. 
19 Unused backup plasma, serum, and urine PK/biomarker samples may also be used for additional exploratory biomarkers analysis. 
 
 


RGLS8429 
Protocol RGLS8429-01 v3.0 
 
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TABLE OF CONTENTS 
TITLE PAGE ...................................................................................................................................1 
SPONSOR SIGNATURE PAGE ....................................................................................................4 
SITE PRINCIPAL INVESTIGATOR SIGNATURE PAGE ..........................................................5 
PROTOCOL SYNOPSIS.................................................................................................................6 
1. 
INTRODUCTION ......................................................................................................18 
1.1. 
Disease Background ...................................................................................................18 
1.2. 
miR-17 and ADPKD ...................................................................................................18 
1.3. 
Nonclinical Pharmacokinetics, Pharmacology, and Toxicology ................................19 
1.4. 
Dose Rationale ............................................................................................................20 
2. 
STUDY OBJECTIVES, ENDPOINTS, AND ASSESSMENTS ...............................21 
2.1. 
Study Objectives .........................................................................................................21 
2.2. 
Study Endpoints ..........................................................................................................21 
2.3. 
Study Assessments and Procedures ............................................................................22 
2.3.1. 
Demographics, Medical History and Concomitant Medications ................................22 
2.3.2. 
Physical Examinations ................................................................................................22 
2.3.3. 
eGFR Calculation .......................................................................................................22 
2.3.4. 
Height and Weight Measurements ..............................................................................22 
2.3.5. 
Vital Signs ..................................................................................................................23 
2.3.6. 
ECGs and Holter Monitoring ......................................................................................23 
2.3.7. 
Safety Laboratory Tests ..............................................................................................23 
2.3.8. 
Pregnancy Test ............................................................................................................24 
2.3.9. 
Urine Drug and Alcohol Screening Test ....................................................................25 
2.3.10. 
HBsAg, HCV Antibody, and HIV-1/2 Antibodies Screening Tests ..........................25 
2.3.11. 
Scale for the Assessment and Rating of Ataxia (SARA) ...........................................25 
2.3.12. 
Pharmacokinetic Samples ...........................................................................................25 
2.3.13. 
Serum Biomarkers ......................................................................................................25 
2.3.14. 
Urine Biomarkers ........................................................................................................26 
2.3.15. 
Exploratory Biomarkers ..............................................................................................26 
3. 
STUDY DESIGN .......................................................................................................27 
3.1. 
Overview of Study Design ..........................................................................................27 
3.2. 
Dose Escalation and Stopping Rules ..........................................................................27 


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3.3. 
Study Duration for Subjects ........................................................................................28 
3.4. 
End of Trial .................................................................................................................28 
4. 
STUDY POPULATION .............................................................................................29 
4.1. 
Number of Subjects ....................................................................................................29 
4.2. 
Inclusion Criteria ........................................................................................................29 
4.3. 
Exclusion Criteria .......................................................................................................30 
5. 
STUDY VISITS ..........................................................................................................32 
5.1. 
Screening Period (Day -28 to Day -1) ........................................................................32 
5.2. 
Treatment Period (Day 1 and Day 2) ..........................................................................33 
5.3. 
Follow-Up Period (Day 3 to Day 29) .........................................................................33 
5.3.1. 
Follow-Up Visit (Day 8) .............................................................................................33 
5.3.2. 
Follow-Up Visit (Day 15) ...........................................................................................34 
5.3.3. 
Follow-Up Visit (Day 22) ...........................................................................................34 
5.3.4. 
Follow-Up / End of Study [EOS] Visit (Day 29) .......................................................35 
5.4. 
Unscheduled Visits .....................................................................................................35 
6. 
STUDY DRUG ...........................................................................................................36 
6.1. 
RGLS8429 ..................................................................................................................36 
6.2. 
Placebo ........................................................................................................................36 
6.3. 
Storage of Study Drug ................................................................................................36 
6.4. 
Subject Identification and Randomization ..................................................................36 
6.5. 
Preparation and Dispensing ........................................................................................37 
7. 
CONCOMITANT MEDICATIONS AND PROCEDURES ......................................38 
7.1. 
Permitted Concomitant Medications and Procedures .................................................38 
7.2. 
Prohibited Concomitant Medications and Procedures ................................................38 
8. 
DATA ANALYSIS / STATISTICAL METHODS ....................................................39 
8.1. 
General Considerations ...............................................................................................39 
8.2. 
Sample Size Determination ........................................................................................39 
8.3. 
Analysis Populations, Subgroups, and Covariates .....................................................39 
8.3.1. 
Analysis Populations ..................................................................................................39 
8.4. 
Final Analysis .............................................................................................................39 
8.4.1. 
Disposition ..................................................................................................................40 
8.4.2. 
Demographic and Baseline Characteristics ................................................................40 
8.4.4. 
Pharmacokinetic Analysis ..........................................................................................41 


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8.4.5. 
Biomarker Analysis ....................................................................................................41 
9. 
ADVERSE EVENTS ..................................................................................................42 
9.1. 
Monitoring, Recording and Reporting of Adverse Events .........................................42 
9.2. 
Evaluation of Adverse Events ....................................................................................42 
9.2.1. 
Seriousness .................................................................................................................42 
9.2.2. 
Severity/Intensity ........................................................................................................43 
9.2.3. 
Causality .....................................................................................................................44 
9.2.4. 
Duration ......................................................................................................................44 
9.2.5. 
Action Taken ..............................................................................................................44 
9.2.6. 
Outcome ......................................................................................................................44 
9.3. 
Abnormal Laboratory Values .....................................................................................45 
9.4. 
Pregnancy ...................................................................................................................45 
9.4.1. 
Male Subjects ..............................................................................................................46 
9.5. 
Reporting of Serious Adverse Events .........................................................................46 
9.6. 
Safety Queries .............................................................................................................46 
9.7. 
Expedited Reporting of Adverse Events .....................................................................46 
9.8. 
Study Pharmacovigilance Group Contact Information ...............................................47 
10. 
DISCONTINUATIONS .............................................................................................48 
11. 
EMERGENCY PROCEDURES ................................................................................49 
11.1. 
Emergency Contact .....................................................................................................49 
12. 
REGULATORY CONSIDERATIONS ......................................................................50 
12.1. 
Good Clinical Practice ................................................................................................50 
12.2. 
Investigator Responsibilities .......................................................................................50 
12.3. 
Subject Information and Informed Consent ...............................................................50 
12.4. 
Confidentiality ............................................................................................................51 
12.5. 
Protocol Amendments ................................................................................................51 
12.6. 
Institutional Review Board/Independent Ethics Committee Review and Approval ..51 
12.7. 
Ongoing Information for Institutional Review Board/ Ethics Committee .................52 
12.8. 
Termination of the Study ............................................................................................52 
13. 
DATA HANDLING AND RECORDKEEPING .......................................................53 
13.1. 
Data/Documents .........................................................................................................53 
13.2. 
Data Management .......................................................................................................53 
13.3. 
Record Retention ........................................................................................................53 


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14. 
QUALITY CONTROL AND QUALITY ASSURANCE .........................................55 
14.1. 
Study Monitoring and Source Data Verification ........................................................55 
14.2. 
Audits and Inspections ................................................................................................55 
15. 
PUBLICATIONS .......................................................................................................56 
16. 
REFERENCES ...........................................................................................................57 
APPENDIX A. TABLE OF ABBREVIATIONS .......................................................................58 
APPENDIX B. SCALE FOR THE ASSESSMENT AND RATING OF ATAXIA (SARA) ....61 
 
LIST OF TABLES 
Table 1: 
Schedule of Assessments and Procedures ..................................................................12 
Table 2: 
Safety Laboratory Tests ..............................................................................................24 
 


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1. 
INTRODUCTION  
1.1. 
Disease Background  
ADPKD, the most common form of polycystic kidney disease (PKD), is a monogenic systemic disease 
driven mostly by mutations in either PKD1 or PKD2 genes, leading to reduced expression and/or 
function of their encoded-proteins polycystin-1 (PC1) and/or polycystin-2 (PC2), excessive proliferation 
of the renal tubular epithelium and formation of multiple cysts (Torres, 2009; Chapman, 2015). Indeed, 
PC1 and PC2 levels in patients’ urinary exosomes were shown to correlate inversely with the height-
adjusted total kidney volume (Hogan, 2015). ADPKD is characterized by slowly progressive, bilateral 
enlargement of the fluid-filled polycystic kidneys, with kidney volume increasing long before detectable 
decline of kidney function. In addition to renal cysts, extrarenal cysts can also develop in the liver, 
pancreas, seminal vesicles, and arachnoid. Typical symptoms include flank pain, hematuria, proteinuria, 
renal colic, urinary tract infection, hypertension, and intracranial aneurysms. Fifty percent of ADPKD 
patients develop end-stage renal disease by the age of 60, accounting for 5% of all end-stage renal 
disease patients in the United States (US) requiring renal replacement therapy such as dialysis or 
transplant (Chebib, 2016). 
1.2. 
miR-17 and ADPKD 
PKD1 and PKD2 genes, which are central to ADPKD pathogenesis, are both direct target genes of miR-
17 (Patel, 2013). Increased miR-17 expression has been detected in kidney samples from both mouse 
ADPKD models and human ADPKD patients (Hajarnis, 2017). Kidney-specific overexpression of miR-
17~92 repressed Pkd1 and Pkd2 expression and produced kidney cysts in mice (Patel, 2013), whereas 
genetic knockdown of miR-17~92 attenuated disease progression in multiple mouse models of PKD 
(Patel, 2013 and Hajarnis, 2017). Importantly, inhibition of miR-17 has been shown to increase PC1 and 
PC2 levels and reduce cyst growth in primary human ADPKD cyst cultures (Hajarnis, 2017 and Lee, 
2019), whereas re-expression of PC1 and PC2 has been shown to rapidly reverse total kidney volume, 
cell proliferation, fibrosis, and inflammation in mouse models of ADPKD (Dong, 2021). Therefore, 
preferential targeting of the miR-17 family of miRNAs in the kidney is an attractive therapeutic 
approach to treat ADPKD. 
Nonclinical pharmacology studies using anti-miR-17 oligonucleotides including the first-generation 
investigational product, RGLS4326, demonstrated that miR-17 inhibition confers therapeutic efficacy in 
multiple mouse models of PKD in vivo and human ADPKD donor primary cyst culture samples in vitro 
(Hajarnis, 2017 and Lee, 2019). Moreover, miR-17 inhibition has been shown to improve expression of 
pathogenic gene networks and pathways in PKD, including de-repression of messenger RNA (mRNA) 
of Pkd1 and Pkd2 genes in mouse models of ADPKD. The coordinated alteration of the polycystic 
kidney transcriptome towards a normal kidney profile following miR-17 inhibition exemplifies the 
potential of an anti-miR-17 oligonucleotide as a disease-modifying treatment for ADPKD (Lee, 2019). 
Importantly, recent data from the first cohort of a Phase 1b clinical trial (NCT04536688) demonstrated 
clinical evidence that treatment with RGLS4326 increased urinary PC1 and PC2 level (Lee, 2021) in 


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patients with ADPKD, likely through inhibition of miR-17 in the kidney. These results imply ongoing 
cis-repression of PC1 and PC2 by the overexpressed miR-17 in ADPKD patients prior to treatment with 
anti-miR-17, further validating miR-17 as a therapeutic target for ADPKD. 
1.3. 
Nonclinical Pharmacokinetics, Pharmacology, and Toxicology 
The following is a brief summary of the Nonclinical information. Please see the RGLS8429 
Investigator’s Brochure for a more detailed presentation of this information.  
In nonclinical ADPKD efficacy studies, RGLS8429 treatment reduced disease progression in both 
Pkd1F/RC and Pcy/DBA mouse models. In the Pkd1F/RC mouse model, RGLS8429 caused a significant 
reduction in kidney weight-to-body weight ratio, serum blood urea nitrogen (BUN), kidney cyst index, 
and kidney cyst proliferation. In the Pcy/DBA mouse model, RGLS8429 reduced kidney weight-to-body 
weight ratio significantly. In addition, the combination of RGLS8429 and tolvaptan demonstrated a 
greater reduction in disease progression than either drug alone in the Pcy/DBA mouse model. 
Based on in silico bioinformatics analysis and in vitro receptor binding studies, RGLS8429 appears to 
have low potential for off-target effects towards ribonucleic acid (RNA) and receptor targets. 
RGLS8429 had no effects on cardiovascular, respiratory, and central nervous system function at 
supraclinical concentrations and dose levels in animals. RGLS8429 had no meaningful effect on the 
human ether-à-go-go-related gene (hERG) potassium channels in vitro. 
In nonclinical PK and toxicokinetic studies, SC administration of RGLS8429 demonstrated rapid 
absorption (time to maximum concentration [Tmax] 1-2 hr) followed by relatively rapid plasma clearance.  
The plasma t1/2 in mice (~ 2 hr) whereas t1/2 ranged between 6 -7 hr in monkeys. 
Plasma exposure increased in a generally dose-proportional manner across all dose levels, and no 
meaningful accumulation was observed in mice or monkeys. RGLS8429 is highly bound to plasma 
protein in mice, monkeys and humans, monkeys, and mice (79.1% ,86.6%, and 81.6 %, respectively). 
RGLS8429 distributes rapidly to the kidney and liver, is metabolically stable with only trace amounts of 
metabolites measured, and is excreted rapidly in the urine, primarily in the first 24 hours after dosing. Of 
note, the decreased renal function present in patients with ADPKD is expected to decrease the renal 
clearance of RGLS8429, and therefore, will be monitored closely in clinical trials. 
Based upon in vitro studies, RGLS8429 has little potential for PK interactions with other drugs via 
either cytochrome P450 enzymes or transporters. In Pcy/DBA mouse model, co-administration of 
RGLS8429 and tolvaptan had no significant effect (defined as <2-fold change) on tolvaptan PK in 
plasma and RGLS8429 PK in kidney and liver. 
Toxicity studies of RGLS8429 completed to date demonstrate that RGLS8429 is not acutely toxic at up 
to 2000 mg/kg in mice.  
In a 13-week Good Laboratory Practice (GLP) toxicity study in CD-1 mice evaluating 60, 300 and 600 
mg/kg weekly of RGLS8429, the no observed adverse effects level (NOAEL) was determined to be 60 
mg/kg, with dose-limiting hepatotoxicity at higher dose levels. Minimal single-cell hepatocellular 
necrosis with 2 to 3-fold elevations in ALT and AST were observed at 600 mg/kg; in addition, one 


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mouse at 300 mg/kg had single-cell hepatocellular necrosis in the absence of elevations in ALT or AST 
at that dose level. Other findings at 300 mg/kg included: decreased thymic lymphocytes with 
corresponding decreased thymus size and weight, and mild decreased red cell mass with compensatory 
hematopoiesis in the spleen and bone marrow, both of which were considered to be anti-miR-17 effects. 
Additional changes at 300 mg/kg included transient nose/muzzle swelling post-dose, decreased serum 
cholesterol, karyomegaly and increased mitotic figures of hepatocytes in the liver (correlating with 
increased liver weight), and increased foamy macrophage infiltration in the injection site(s), all of which 
were considered non-adverse. 
In a 13-week GLP toxicity study in cynomolgus monkeys evaluating 15, 75 and 150 mg/kg weekly of 
RGLS8429, highest dose of RGLS8429 (150 mg/kg) was well tolerated and was determined to be the 
NOAEL. The following findings, all of which were considered to be non-adverse, include: minimal 
increases in fibrinogen, minimal but transient increases in activated thromboplastin time (APTT), dose-
related dark red discoloration at the injection site with microscopic findings of fibrosis, macrophage 
infiltration, perivascular inflammation, and hemorrhage, vacuolated macrophages in the lymph nodes, 
and basophilic granules within Kupffer cells in the liver.  
SC administration of the clinical formulation (150 mg/kg in water for injection) was well tolerated in 
rabbits. RGLS8429 has shown no potential for genotoxicity in two in vitro studies (Ames test and 
micronucleus assay).  
RGLS8429 did not cause overt cytotoxicity and showed no evidence of mitochondrial toxicity. Although 
certain oligonucleotides have exhibited the propensity to cause thrombocytopenia in animals and/or 
humans, in vitro studies suggest that RGLS8429 has minimal potential for adverse effects on platelets, 
which is consistent with the absence of adverse effects in mice and monkeys. 
In addition, RGLS8429 did not induce genes involved in inflammation, consistent with the low pro-
inflammatory effects observed. 
1.4. 
Dose Rationale 
The starting dose and dose levels selected for first-in-human dosing are based on the NOAELs and 
toxicity profiles in the GLP 13-week toxicity studies of RGLS8429. The NOAELs in mice and monkeys 
were 60 mg/kg and 150 mg/kg, respectively, for human equivalent doses (HED) of 5 and 48, 
respectively. Based on the HED in the more sensitive species (mouse) and using a safety factor of 
approximately 10 (in accordance with FDA Guidance for Industry, Estimating the Maximum Safe 
Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers [July 2005]), the 
starting dose in this study is 0.5 mg/kg and maximum dose will not exceed 5 mg/kg. 


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2. 
STUDY OBJECTIVES, ENDPOINTS, AND ASSESSMENTS 
2.1. 
Study Objectives 
Primary Objective 
x To assess the safety and tolerability of single ascending SC doses of RGLS8429  
Secondary Objectives 
x To identify DLT and to determine the MTD of a single SC dose of RGLS8429 
x To characterize the PK properties of RGLS8429 
Exploratory Objective 
x To characterize multiple potential renal biomarkers 
2.2. 
Study Endpoints 
Primary Endpoint 
x Incidence of adverse events (AEs) and change in safety laboratory test results, vital signs, 
SARA, and ECG parameters and Holter monitoring over time 
Secondary Endpoints 
x Incidence of DLT and determination of MTD 
x PK parameters of RGLS8429 including: 
o Maximum observed concentration (Cmax)  
o Time to maximum observed concentration (Tmax) 
o Area under the concentration–time curve up to 24 hr post dose (AUC0-24) 
o Area under the concentration–time curve up to the last quantifiable concentration 
(AUC0-t) 
o Area under the concentration–time curve extrapolated to infinity (AUCinf)  
o Half-life (t½)  
o Apparent clearance (CL/F)  
o Volume of distribution (Vz/F) 
o Fraction of unchanged RGLS8429 excreted in the urine (fe) 
o Total amount of unchanged RGLS8429 excreted in the urine (Ae) 
Exploratory Endpoints 
x Characterize multiple potential renal biomarkers in urine (e.g., PC2, NGAL, KIM-1, MCP-1, 
B2M, and CLU) and in serum (e.g., Cystatin-C, IGFALS, Alb, CT-proAVP, N-acetyl-1-
methylhistidine, and others) 


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2.3. 
Study Assessments and Procedures 
Informed consent will be obtained prior to performing any study procedures.  
2.3.1. 
Demographics, Medical History and Concomitant Medications 
Subject demographic data collection, a review of each subject’s medical history, and prior/current 
concomitant medications and medical procedures will be conducted according to the Schedule of 
Assessments and Procedures (see Table 1) to ensure all study entry criteria are met. 
2.3.2. 
Physical Examinations 
Physical examinations may be conducted by a physician or another medically qualified individual such 
as a physician's assistant or a nurse practitioner according to the Schedule of Assessments and 
Procedures (see Table 1). All physical examinations must include a complete neurologic exam. A full 
physical examination will include general appearance, head, ears, eyes, nose, mouth, skin, heart, lung, 
lymph nodes, and the gastrointestinal, musculoskeletal, respiratory, and neurological systems. The 
limited physical examination will be focused on general appearance, the respiratory, the neurological, 
and cardiovascular systems, and subject-reported symptoms.  
2.3.3. 
eGFR Calculation 
The Investigator will calculate the estimated glomerular filtration rate (eGFR) using the 2021 (chronic 
kidney disease epidemiology collaboration equation) CKD-EPI Creatinine, Age, Sex Equation (Inker, 
2021) during Screening: 
142 x min(Scr/k,1)Į x max(Scr/k,1)-1.200 0.9938age x 1.012 [if female]  
Scr is serum creatinine 
k is 0.7 for females and 0.9 males 
ĮLV-0.241 for females and -0.302 for males 
min indicates the minimum of Scr/k or 1 
max indicates the maximum of Scr/k or 1 
2.3.4. 
Height and Weight Measurements 
Height will be recorded only during Screening. 
Weight will be measured according to the Schedule of Assessments and Procedures (see Table1). For 
measuring weight, a scale with appropriate range and resolution should be used and must be placed on a 
stable, flat surface. Subjects must remove shoes, bulky layers of clothing, and jackets so that only light 
clothing remains. Subjects must also remove the contents of their pockets and should remain still during 
measurement of weight.  


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2.3.5. 
Vital Signs 
Body temperature, seated systolic and diastolic blood pressure (BP), heart rate (HR), and respiratory rate 
(RR) will be measured at times specified in the Schedule of Assessments and Procedures (see Table 1).  
Body temperature may be measured orally, via the external auditory canal, or via the temporal artery 
(forehead). Temperature should be measured in a consistent manner for each subject throughout the 
entire study. 
The use of an automated device for measuring BP and HR is acceptable. For BPs, a properly sized and 
calibrated BP cuff should be used for each measurement. Subjects should be seated for approximately 5 
minutes before the BP is measured. The BP should be measured with the subject’s feet flat on the floor, 
back supported, and arm supported at the level of the heart. The BP should be recorded to the nearest
millimeters of mercury (mmHg).  
When the timing of these measurements coincides with a blood sample collection, obtain the vital signs, 
and then collect the blood sample.  
2.3.6. 
ECGs and Holter Monitoring 
ECGs and Holter monitoring should be obtained at times specified in the Schedule of Assessments and 
Procedures (see Table 1). 
The Investigator or a medically qualified designee should review all ECGs obtained during the study. 
Consultation with a cardiologist should be obtained if necessary. Record new or worsening clinically 
significant abnormalities as an AE. Post-treatment ECG results should be compared with the pre-dose 
ECG assessments. 
Holter monitoring should be performed by the Investigator or a medically qualified designee at times 
specified in the Schedule of Assessments and Procedures (see Table 1). The Holter data will be analyzed 
centrally, and the independent central reader will remain blinded to treatment group. Throughout the 
duration of Holter monitoring, other procedures and SARA assessment may occur. At the specified 
timepoints, the procedures will be performed in this order: ECG, vital signs, PK and/or other blood 
collection, and then followed by the SARA assessment. Details and procedures will be provided in the 
Holter Monitoring Manual. 
2.3.7. 
Safety Laboratory Tests 
The Safety Laboratory Tests listed in Table 2 should be obtained at the times specified in the Schedule 
of Assessments and Procedures (see Table 1). Detailed collection, processing, storage, and shipment 
instructions will be provided in a Laboratory Manual. 


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Table 2: 
Safety Laboratory Tests 
Hematology – Complete Blood Count 
x Hemoglobin (Hb) 
x Hematocrit (Hct) 
x Red Blood Cell (RBC) Count 
x Mean Corpuscular Volume (MCV) 
x Mean Corpuscular Hemoglobin (MCH) 
x Mean Corpuscular Hemoglobin Concentration 
(MCHC) 
x Platelet Count 
x White Blood Cell (WBC) Count and 
Differential 
x Absolute Neutrophil Count 
x Absolute Lymphocyte Count 
x Absolute Monocyte Count 
x Absolute Eosinophil Count 
x Absolute Basophil Count 
Chemistry – Metabolic Panel 
x Glucose 
x Calcium 
x Albumin (Alb) 
x Total Protein  
x Sodium 
x Potassium 
x Bicarbonate (Total CO2) 
x Chloride 
x Blood Urea Nitrogen (BUN) 
x Uric Acid 
x Creatine kinase (CK) 
x Phosphate 
x Lactate Dehydrogenase (LDH) 
x Blood Urea Nitrogen (BUN) 
x Creatinine (Scr) 
x Alkaline Phosphatase (ALP) 
x Alanine Aminotransferase (ALT) 
x Aspartate Aminotransferase (AST) 
x Gamma-Glutamyl Transferase (GGT) 
x Total Bilirubin  
x Direct Bilirubin 
Urinalysis a 
x Specific Gravity 
x pH 
x Protein  
x Glucose 
x Ketones 
x Blood (Hemoglobin) 
x Leukocyte Esterase Nitrite 
x Bilirubin 
x Urobilinogen 
x Microscopic Examination b 
Coagulation 
x International Normalized Ratio (INR) 
x Prothrombin Time (PT) 
x Activated Partial Thromboplastin Time (aPTT) 
x Fibrinogen 
Lipids 
x Total cholesterol 
x Triglycerides 
x High-density Lipoprotein (HDL) 
x Low-density Lipoprotein (LDL) 
Unscheduled safety laboratory assessments may be obtained at any time during the study to assess potential safety concerns. 
a 
Urinalysis testing will be conducted using standard commercial test strips (urine dipstick).  
b 
Only if urine dipstick is positive for blood, protein, or leukocyte esterase nitrate. 
2.3.8. 
Pregnancy Test 
For female subjects of childbearing potential, a serum pregnancy test will be performed at Screening. 
Urine pregnancy tests will be performed at the remaining specified visits according to the Schedule of 
Assessments and Procedures (see Table 1). A positive urine pregnancy test will be confirmed with a 
serum pregnancy test. 


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2.3.9. 
Urine Drug and Alcohol Screening Test 
The drug and alcohol screen will include measurement of opiates, cocaine, heroin, phencyclidine, 
amphetamines (including ecstasy), barbiturates, benzodiazepines, cannabinoids, and alcohol according 
to the Schedule of Assessments and Procedures (see Table 1). 
2.3.10. 
HBsAg, HCV Antibody, and HIV-1/2 Antibodies Screening Tests 
All subjects will be tested for HBsAg, HCV antibody, and HIV-1/2 antibodies at Screening. 
2.3.11. 
Scale for the Assessment and Rating of Ataxia (SARA) 
The SARA test is used in this study to detect possible central nervous system impairment by the study 
drug and should be performed according to the Schedule of Assessments and Procedures (see Table 1). 
When timepoints to perform the SARA assessment are near timepoints for other procedures, the 
procedures will be performed in this order: ECG, vital signs, PK and/or other blood collection, and then 
followed by the SARA assessment. The test should be performed by an Investigator who has been 
trained to conduct neurologic evaluations and should ideally be performed by the same individual during 
the study to minimize the variability in how the test is performed and scored. The scale is presented in 
Appendix B. 
2.3.12. 
Pharmacokinetic Samples 
Plasma PK samples will be collected at the times specified in the Schedule of Assessments and 
Procedures (see Table 1).  
All efforts should be made to obtain PK samples at the specified nominal time relative to dosing. Pre-
dose plasma PK sample on Day 1 should be obtained within 60 minutes prior to dosing. Post-dose 
plasma samples should be obtained within the following time margins: collections at 2, 4, 6, and 8 hr 
±15 minutes, 12 hr ±30 minutes, and 24 hr±60 minutes. The exact date and time of collection for each 
sample should be noted on the source document and in the case report form (CRF). Detailed collection, 
processing, storage, and shipment instructions for PK samples will be provided in the Laboratory 
Manual. 
24-hr urine collection for PK analysis will begin immediately after study drug dosing on Day 1 (0-24 
hr). The subject should void immediately before dosing (before the 24-hr urine collection is started) and 
the last void will be timed for 24 hours post-dose. Detailed collection, processing, storage, and shipment 
instructions for PK samples will be provided in the Laboratory Manual. 
2.3.13. 
Serum Biomarkers 
Collect serum samples during Screening and on Day -1 according to the Schedule of Assessments and 
Procedures (see Table 1) for exploratory biomarker analysis (e.g., Cystatin-C, IGFALS, Alb, CT-
proAVP, N-acetyl-1-methylhistidine, and others). Detailed collection, processing, storage, and shipment 
instructions for serum biomarker samples will be provided in the Laboratory Manual. 


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2.3.14. 
Urine Biomarkers 
Collect at least 150 mL (more if possible) of mid-stream free-flow urine for analysis of exploratory 
biomarkers (PC1, and PC2, NGAL, KIM-1, MCP-1, B2M, CLU) during Screening and on Day -1 
according to the Schedule of Assessments and Procedures (see Table 1). These samples must NOT be 
the first urination of the day (to avoid too much debris in the sample). Detailed collection, processing, 
storage, and shipment instructions for urine biomarker samples will be provided in the Laboratory 
Manual. 
2.3.15. 
Exploratory Biomarkers 
Residual plasma, serum, and urine samples (e.g., unused or backup) not utilized in required testing of 
PK/biomarkers will be retained and used to measure the concentrations of multiple potential exploratory 
renal function biomarkers in healthy volunteers. Subjects will provide consent for the use of these 
residual samples for exploratory tests.  


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3. 
STUDY DESIGN 
3.1. 
Overview of Study Design 
In this randomized, double-blind, placebo-controlled Phase 1 study, a single ascending dose of 
RGLS8429 or placebo will be administered via SC injection to healthy volunteers to evaluate the safety, 
tolerability, and PK of RGLS8429. Eight subjects will be randomized 3:1 to receive RGLS8429 or 
placebo in each cohort.  
Cohort 1 
0.5 mg/kg RGLS8429 or placebo 
Cohort 2 
1.0 mg/kg RGLS8429 or placebo 
Cohort 3 
2.0 mg/kg RGLS8429 or placebo 
Cohort 4 
4.0 mg/kg RGLS8429 or placebo 
In each cohort, 6 subjects will be randomized to receive RGLS8429 and 2 will be randomized to receive 
placebo. Enrollment into subsequent cohorts will be initiated after review of safety (AEs and safety 
laboratory tests) from the prior cohort.  
For Cohort 1, the first two subjects will receive RGLS8429 or placebo (randomized such that one 
subject receives RGLS8429 and one receives placebo) and this sentinel group will be monitored for 48 
hr prior to dosing the remaining subjects in the cohort. 
An unblinded pharmacist will be responsible for preparing RGLS8429 or placebo and providing the 
study drug to blinded study personnel for administration according to the randomization procedures.  
Subjects will be admitted to the study center on Day -1, one day prior to the day of study drug 
administration (Day 1), and will be discharged on Day 2, no less than 6 hr after the collection of the 24-
hr PK sample. Subjects will be followed for a total of 4 weeks after dosing and will return to the site for 
follow-up visits weekly from Weeks 1 through 4. The sentinel subjects will remain in the clinic for 48 hr 
after dosing. 
3.2. 
Dose Escalation and Stopping Rules 
Prior to dose escalation, the Sponsor and the Investigator will perform a blinded review of the safety 
data (e.g., AE and safety laboratory tests through Day 15) on all subjects at the current dose level to 
make a dose escalation decision.  
If 2 or more subjects at the same dose level experience the saPHRUVLPLODU*UDGH$(VLQFOXGLQJ
FOLQLFDOO\VLJQLILFDQWODERUDWRU\$(VRULIVXEMHFWH[SHULHQFHVD*UDGH$(RUD6$(MXGJHGWREH
related to study drug, the Sponsor’s Lead Physician will be unblinded to the treatment. Also, if 2 
subjects develop stage 1 AKI or 1 subject develops stage 2 AKI the Sponsor’s Lead Physician will be 
unblinded to the treatment. 
The Sponsor’s Lead Physician will determine whether the nature, severity, or number of event(s) 
warrants: 


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1. Continuing with the study as planned 
2. Continuing with the study, but add additional safety evaluation(s) 
3. Suspend dose escalation and enroll additional subjects at the same dose level 
4. Add an intermediate-dose cohort or adjust the next dose level 
5. Suspend or terminate the study 
3.3. 
Study Duration for Subjects  
Subjects will participate in the study for up to 57 days (Screening up to 28 days, Treatment 1 day, 
Follow-up 28 days). 
3.4. 
End of Trial 
The End of Trial is defined as either the date of the last visit for the last subject to complete the post-
treatment follow-up, or the date of receipt of the last data point from a subject, whichever is the later 
date. 


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4. 
STUDY POPULATION 
4.1. 
Number of Subjects  
Approximately 32 healthy volunteers will be enrolled. Subjects may be replaced at the discretion of the 
Sponsor (e.g., subject noncompliance with the protocol, early withdrawal, missing data, etc.). 
4.2. 
Inclusion Criteria  
Subjects must meet the following criteria to be eligible for study participation: 
1. Male or female, 18 to 55 years of age  
2. Continuous nonsmoker who has not used nicotine-containing products for at least 3 months prior 
to dosing 
3. Body mass index (BMI) 18 to 35 kg/m2 
4. Medically healthy, with no clinically significant medical history in the opinion of the 
Investigator. 
5. Estimated glomerular filtration rate (eGFR) 90 mL/min/1.73 m2 (see Section 2.3.3) 
6. Platelet count within the normal limit 
7. Hemoglobin concentration within the normal limits 
8. Liver function tests (ALT, $67$/3DQGELOLUXELQ>WRWDODQGGLUHFW@PXVWEH the ULN 
9. Female subjects of childbearing potential must not be lactating and must have no plans to 
become pregnant during the course of the study through 28 days after the administration of study 
drug. Female subjects of childbearing potential who are heterosexual must agree to use one of 
the following methods of contraception considered to be highly effective (i.e., results in <1% 
failure rate when used consistently and correctly) from Screening through 28 days after the 
administration of study drug 
a. Intrauterine device (IUD) or intrauterine system (IUS) in place for at least 3 months 
prior to study drug administration  
b. Partner has had a vasectomy. Vasectomy in the partner is considered to be highly 
effective only if the partner is the sole sexual partner of the female subject of 
childbearing potential and had his vasectomy performed 6 months or more prior to 
randomization. 
c. Stable hormonal contraception associated with inhibition of ovulation (with approved 
oral, transdermal, or depot [e.g., NuvaRing, depot injection, etc.] regimen) for at least 
3 months prior to study drug administration 
10. A female subject of non-childbearing potential must have undergone one of the following 
sterilization procedures at least 6 months prior to administration of study drug: 
a. Hysterectomy  
b. Bilateral oophorectomy  
c. Bilateral tubal occlusion 


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d. Bilateral salpingectomy 
or be postmenopausal with no menstrual periods for at least 1 year prior to administration of study 
drug. 
11. Non-vasectomized heterosexual male subjects must agree to use a condom with spermicide or 
abstain from sexual intercourse from Day 1 until 28 days after the administration of study drug. 
(No restrictions are required for a vasectomized heterosexual male provided his vasectomy was 
performed 6 months or more prior to study start. A heterosexual male who has been 
vasectomized less than 6 months prior to study start must follow the same restrictions as a non-
vasectomized heterosexual male.) 
12. Male and female subjects must agree not to donate sperm or preserve eggs (ova), respectively, 
from Day 1 until 28 days after the administration of study drug 
13. Must agree not to donate blood in the 28 days prior to randomization or plasma in the 7 days 
prior to randomization through the EOS visit 
14. Seated blood pressure (systolic/diastolic) is  mmHg and  140/90 mmHg 
15. Seated heart rate is  40 bpm and  99 bpm 
16. Fridericia-corrected QT (QTcF) interval is < 460 msec (males) or < 480 msec (females) and 
without any ECG findings deemed clinically significant by the Investigator 
17. Must understand and consent to the study procedures explained in the informed consent form 
(ICF) and be willing and able to comply with the protocol 
4.3. 
Exclusion Criteria 
Subjects with any of the following will be excluded from participation in the study: 
1. Subject is mentally incapacitated or has significant emotional problems 
2. Any medical condition or social circumstance that, in the opinion of the Investigator, may make 
the subject unlikely to complete the study or comply with study procedures and requirements, or 
may pose a risk to the subject’s safety 
3. History of malignancy, except for successfully treated squamous or basal cell carcinoma skin 
cancer  
4. History or presence of opportunistic infection, in the opinion of the Investigator 
5. History or presence of any clinically significant local infection (e.g., cellulitis, abscess) or 
systemic infection (e.g., septicemia) within 3 months prior to Screening 
6. Fever (body temperature  100.4°F) or symptomatic viral or bacterial infection within 14 days 
prior to Screening, at Screening, at Day -1 check-in, or at Day 1 prior to dosing 
7. History of any clinically significant reaction to previous injection of an oligonucleotide product 
in the opinion of the Investigator 
8. History or presence of alcoholism or drug abuse within the past 2 years prior to Screening 
9. Positive urine drug or alcohol results  
10. +DVSrotein on urine dipstick  


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11. Positive results at Screening for human immunodeficiency virus (HIV), hepatitis B surface 
antigen (HBsAg), or hepatitis C virus (HCV) antibody 
12. Positive urine cotinine  
13. Unable to refrain from, or anticipates the use of, any drug, including prescription and 
non-prescription medications and herbal remedies, from 14 days prior to dosing until completion 
of the EOS visit, except as approved by the Investigator  
14. History of any major surgical procedure within 3 months prior to dosing (excluding minor 
cosmetic surgery or minor dental procedures) 
15. Subjects with tattoo(s) or scarring at or near the site of SC injection or any other condition that 
may interfere with injection site examination(s), in the opinion of the Investigator 
16. Participation in another clinical trial and/or exposure to any investigational drug or approved 
therapy for investigational use within 28 days or 5 half-lives of the investigational drug’s dosing, 
whichever is longer, prior to dosing. The 28-day and 5-half-life windows will be calculated from 
the date of the last dosing in the previous study to Day 1 of the current study. 


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5. 
STUDY VISITS 
Study visits and procedures will be performed as outlined in Table 1. Additional details on the study 
assessments and procedures are given in Section 2.3. The study will consist of Screening, Treatment, 
and Follow-up periods. Additional visits may be scheduled to evaluate an abnormal laboratory value or 
reported AE. For each study subject, written informed consent will be obtained prior to any protocol-
related activities. After signing of the ICF, completion of all Screening assessments, and confirmation of 
eligibility, subjects will be randomized for the study. 
5.1. 
Screening Period (Day -28 to Day -1) 
Subjects will be screened during the 28 days prior to administration of study drug (Day 1) to confirm 
that they meet the entry criteria for the study. Some of the procedures must occur on Day -1 after the 
subject is admitted to the clinic. Subjects who fail Screening (i.e., do not meet entry criteria) may be 
rescreened with the approval of the Sponsor’s Lead Physician.  
The following procedures are to be performed during Screening and/or Day -1: 
x Informed consent 
x Demographics (date of birth [ddmmmyyyy], sex, race, and ethnicity) 
x Medical history 
x Complete physical examination; must include a complete neurologic examination 
x Weight (Screening and Day -1) and height (Screening); calculate BMI using Screening 
values; Day -1 weight will be used for dosing 
x Vital signs (Screening and Day -1) 
x 12 -lead ECG (Screening); Print and review 
x Safety laboratory tests (Screening and Day -1): 
o Hematology 
o Chemistry 
o Urinalysis 
o Lipids 
x Coagulation tests (Day -1) 
x Pregnancy test, if applicable (serum at Screening; urine at and Day -1) 
x Urine drug and alcohol screen (Screening and Day -1) 
x HBsAg, HCV, HIV-1/2 (Screening) 
x Serum biomarker sample collection (Screening and Day -1) 
x Urine biomarker sample collection (Screening and Day -1) 
x Inclusion/exclusion criteria review (Screening and Day -1) 
x Randomization after enrollment confirmed (Day -1) (see Section 6.4) 


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x Concomitant medications and procedures (Screening and Day -1) 
x Admission to clinic (Day -1) 
5.2. 
Treatment Period (Day 1 and Day 2) 
The following procedures are to be performed on Day 1 and/or Day 2: 
x Limited physical examination (pre-dose Day 1 and 24 hr post-dose Day 2) 
x Vital signs (Day 1 pre-dose and 1, 2, 4, and 8 hr post-dose; Day 2, 24 hr post-dose) 
x Holter monitoring should be initiated 1 hr pre-dose and continue for approximately 30 
minutes after the 24-hr PK sample is obtained (Day 1 and Day 2).  
x Print and review ECG tracings pre-dose and 4, 6, 12, and 24 hr post-dose. 
x SARA assessment (Day 1 pre-dose and 4-6 hr post-dose; Day 2, 24 hr post-dose) 
Note: The pre-dose ECG tracing and SARA procedures may be performed prior to the 
start of Holter monitoring at the 1 hr pre-dose timepoint on Day 1. 
x Safety laboratory tests (Day 2, 24 hr post-dose): 
o Hematology 
o Chemistry 
o Urinalysis 
o Lipids 
x Coagulation test (Day 1, 4 hr post-dose and Day 2, 24 hr post-dose) 
x AE monitoring (Day 1 through Day 2) 
x Concomitant medications and procedures (Day 1 through Day 2) 
x Study drug administration (Day 1 at Time Zero) 
x PK plasma samples (Day 1 pre-dose and 2, 4, 6, 8, and 12 hr post-dose; Day 2, 24 hr post-
dose. Pre-dose plasma PK sample on Day 1 should be obtained within 60 minutes prior to 
dosing. Post-dose plasma samples should be obtained within the following time margins: 
collections at 2, 4, 6, and 8 hr±15 minutes, 12 hr±30 minutes, and 24 hr±60 minutes. 
x 24-hr urine collection on Day 1 and Day 2 for PK analysis will begin immediately after study 
drug dosing on Day 1 (0-24 hr). The subject should void immediately before dosing and at 
the end of the 24-hr collection period. 
x Discharge from clinic (Day 2) no less than 6 hr after the 24-hr PK sample collection 
5.3. 
Follow-Up Period (Day 3 to Day 29) 
5.3.1. 
Follow-Up Visit (Day 8) 
The following procedures are to be performed on Day 8: 
x Limited physical examination; must include a complete neurologic examination 


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x Weight 
x Vital signs 
x Safety laboratory tests: 
o Hematology 
o Chemistry 
o Urinalysis 
x AEs since the last visit 
x Concomitant medications and procedures since last visit 
5.3.2. 
Follow-Up Visit (Day 15) 
The following procedures are to be performed on Day 15: 
x Limited physical examination; must include a complete neurologic examination 
x Vital signs 
x Safety laboratory tests: 
o Hematology 
o Chemistry 
o Urinalysis 
o Lipids 
x AEs since the last visit 
x Concomitant medications and procedures since last visit 
x Plasma PK sample 
5.3.3. 
Follow-Up Visit (Day 22) 
The following procedures are to be performed on Day 22: 
x Limited physical examination; must include a complete neurologic examination 
x Vital signs 
x Safety laboratory tests: 
o Hematology 
o Chemistry 
o Urinalysis 
x AEs since the last visit 
x Concomitant medications and procedures since last visit 


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5.3.4. 
Follow-Up / End of Study [EOS] Visit (Day 29) 
The following procedures are to be performed on Day 29; or if the subject withdraws from the study 
early, these procedures should be performed: 
x Complete physical examination; must include a complete neurologic examination 
x Weight 
x Vital signs 
x Safety laboratory tests 
o Hematology 
o Chemistry 
o Coagulation 
o Urinalysis 
o Lipids 
x Urine pregnancy test, if applicable 
x Urine drug and alcohol screen 
x SARA Assessment 
x AEs since the last visit 
x Concomitant medications and procedures since the last visit 
5.4. 
Unscheduled Visits 
Unscheduled visits may be conducted at the Investigator’s discretion and/or when repeat of laboratory 
assessments is needed. In addition, the following procedures will occur at any unscheduled visits:  
x Adverse event evaluation  
x Concomitant medications and procedures since last visit will be reviewed 
x Additional assessments may be performed, as determined by the Investigator 
 


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6. 
STUDY DRUG 
6.1. 
RGLS8429 
RGLS8429 injection will be provided in 2 mL clear glass vials containing sufficient volume to extract 
the labeled volume of 1 mL of 150 mg/mL of RGLS8429 in 0.3% saline. The solution is clear and 
colorless to pale yellow. 
6.2. 
Placebo 
Placebo injection will be provided in 2 mL clear glass vials containing sufficient volume to extract the 
labeled volume of 1 mL of 1.5 μg/mL of riboflavin in 0.9% sodium chloride. The solution is clear and 
colorless to pale yellow. 
6.3. 
Storage of Study Drug 
Site staff must ensure that all study drug is stored in a secured area with controlled access under required 
storage conditions and in accordance with applicable regulatory requirements. To ensure that the 
investigative staff are not unblinded, only unblinded pharmacy staff and an unblinded clinical research 
associate should have access to the unblinded study drug. The clinic and non-pharmacy staff should be 
provided only with blinded syringes for injection to subjects. 
Study drug vials should be stored in their original containers and in accordance with the labels. Study 
drug must be stored in a freezer at -20° ± 5°C. The study drug should be stored in its carton to protect it 
from light until it is removed from the carton to prepare for administration.  
The site must be capable of measuring and documenting the daily minimum and maximum temperatures 
for all freezers used to store study drug. Temperature monitoring information should be captured from 
the time of study drug receipt throughout the duration of the study. A site procedure that ensures active 
evaluation for temperature excursions should be in place and appropriate documentation of temperature 
monitoring must be available. The minimum and maximum temperature should be checked each 
business day to confirm that no excursion occurred, and the site should have the capability to view the 
minimum/maximum temperature for all nonworking days upon return to normal operations. The 
temperature monitoring device and freezer used to store study drug should be inspected regularly to 
ensure that they are maintained in working order.  
Any excursions from the labeled storage conditions should be reported to Regulus or designee upon 
discovery. In the event of an excursion, the site should ensure that the product is returned to the storage 
conditions described in the labeling as soon as possible. Deviations from the storage requirements, 
including any actions taken, must be documented, and reported to Regulus or designee.  
Receipt of study drug, opening and closing the freezer, and other routine handling operations where the 
study drug is briefly out of the temperature range described in the labeling will not be considered 
excursions.  
6.4. 
Subject Identification and Randomization 
The Investigator or a designee will assign sequential Screening identification numbers to the subjects as 
they are screened for the study. Subjects will then be assigned different sequential subject identification 
numbers as they are enrolled in the study. Details regarding subject identification procedures will be 
provided in the Study Reference Manual. 


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Regulus or designee will provide a randomization schedule and eligible subjects will be randomized 
sequentially to receive the study drug assigned to the corresponding randomization number. Subjects 
may be randomized once all Screening and Day -1 procedures have been completed and eligibility has 
been confirmed by the Investigator. Details regarding randomization procedures will be provided in the 
Study Reference Manual. 
6.5. 
Preparation and Dispensing 
Study drug will be prepared by an unblinded pharmacist or other qualified and trained personnel at the 
site as delegated on the delegation log. The blinded study drug will be provided to the blinded study 
personnel for administration according to the randomization procedures.  
The study drug vials are for single use only. The Study Reference Manual will provide detailed 
instructions on how to prepare the study drug for administration.  
6.6. 
Administration 
Study drug will be administered via SC injection as a bolus in the anterior abdominal wall in accordance 
with the standard-of-care procedures at the site. Study drug should be administered by the Investigator, 
or another qualified and trained site staff member as delegated on the delegation log. Because the 
volume of study drug will vary considerably depending on the dose level, the following guideline must 
be followed: the maximum volume for each injection must not exceed 2 mL (e.g., a 6-mL dose would 
require three 2 mL injections in the abdomen). The Study Reference Manual will provide detailed 
instructions on how to prepare and administer the study drug.  
6.7. 
Study Drug Accountability 
The site must maintain adequate records documenting the receipt, preparation, use, loss, or other 
disposition of the study drug. All study drug will be accounted for using a drug accountability 
form/record.  
6.8. 
Destruction of Study Drug Supplies 
Regulus or designee will provide guidance on the destruction of unused study drug at the site. The 
Investigator must ensure that the materials are destroyed in compliance with applicable environmental 
regulations, institutional policy, and any special instructions provided by Regulus. All study drug 
destruction must be documented adequately. 


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7. 
CONCOMITANT MEDICATIONS AND PROCEDURES 
7.1. 
Permitted Concomitant Medications and Procedures 
A concomitant medication is any drug or substance other than study drug, including over-the-counter 
medications, herbal medications, and vitamin supplements, administered during the subject’s 
participation in this study. A concomitant procedure is any therapeutic intervention (e.g., surgery/biopsy, 
physical therapy) or diagnostic assessment (e.g., blood gas measurement, bacterial cultures) performed 
during the subject’s participation in this study.  
Concomitant medications (and dosing adjustments) and procedures to treat new or existing medical 
conditions or AEs will be permitted. 
All medications and procedures used in the 3 months before Randomization and the subsequent 
concomitant medications and procedures received/performed during the study must be recorded.  
7.2. 
Prohibited Concomitant Medications and Procedures 
Not applicable as long as guidelines for permitted concomitant medications and procedures are 
followed. 


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8. 
DATA ANALYSIS / STATISTICAL METHODS 
8.1. 
General Considerations 
The primary objective of this study is to assess the safety and tolerability of RGLS8429. Secondary 
objectives include characterizing the PK of RGLS8429 and to characterize several exploratory urine and 
serum biomarkers.  
Tabular summaries and analysis results will be generated by dose level for RGLS8429 and placebo 
(pooled across all cohorts). Descriptive statistics by dose level and treatment group will be tabulated by 
visit. For continuous endpoints, the descriptive statistics will include the number of observations, mean, 
median, standard deviation, coefficient of variation (%CV), first and third quartiles, minimum and 
maximum. Geometric mean and geometric %CV will be presented where appropriate. For categorical 
endpoints, the number and percentage of subjects will be summarized by dose level and treatment group. 
Details of all statistical methods will be provided in the Statistical Analysis Plan. 
8.2. 
Sample Size Determination 
Formal sample size calculations were not performed for this study. The dose cohort size is 8 subjects 
randomized 3:1 to RGLS8429 versus placebo. The size of each dose cohort was chosen to provide 
sufficient information to allow assessment of the safety and tolerability of RGLS8429. 
8.3. 
Analysis Populations, Subgroups, and Covariates 
8.3.1. 
Analysis Populations 
Safety Population 
The Safety Population consists of all subjects who received a dose of study drug.  
Pharmacokinetic Full Population 
The PK Full Population consists of all subjects who received a dose of RGLS8429 and have at least 1 
concentration data point. Subjects will be analyzed based on dose level received. 
Pharmacokinetic Evaluable Population 
The PK Evaluable Population consists of all subjects who received a dose of RGLS8429 and have at 
least one PK parameter that can be reliably estimated. Subjects will be analyzed based on dose level 
received. 
Biomarkers Population 
The PD Population consists of all randomized subjects who have at least one Screening and/or pre-dose 
value for the biomarker parameters.  
8.4. 
Final Analysis 
The final analysis of the study will be performed when all subjects have received study drug and been 
followed for 4 weeks.  


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8.4.1. 
Disposition 
The number and percent of subjects enrolled, receiving assigned treatment, completed assigned 
treatment, and completing study will be tabulated by treatment cohort. Subjects who withdraw from 
study drug and/or study prematurely will be summarized by treatment cohort with the reason for early 
discontinuation.  
8.4.2. 
Demographic and Baseline Characteristics 
Subject demographics (age, sex, race, ethnicity) and baseline characteristics will be summarized 
descriptively by treatment cohort and for all subjects based on the Safety Population. 
8.4.3. 
Safety Analysis 
The Safety Population will be used for all safety analyses. No formal statistical tests will be conducted 
to assess the safety or tolerability of RGLS8429.  
Subject incidence of all TEAEs, all TEAEs related to study drug, and all SAEs will be tabulated by 
system organ class and preferred term. All fatal adverse events, SAEs, and TEAEs leading to withdrawal 
from study drug will also be listed. 
The incidence of DLTs will be summarized by dose level and treatment group. 
The analyses of safety laboratory tests, vital signs, and ECGs will include summary statistics over time 
tabulated by cohort. Similarly, changes from baseline will be summarized. Shifts based on normal 
ranges between the baseline and the worst on-study value will be tabulated by dose level and treatment 
group. 
The results from the central reader assessment of ECGs extracted from the Holter data will be used in all 
statistical analysis. The following intervals will be summarized by treatment group: PR, QRS, RR, HR, 
uncorrected QT, QTcB, and QTcF. Details of additional analyses will be provided in the Statistical 
Analysis Plan. 
The total dose of study drug and the proportion of subjects receiving each dose level will be summarized 
using descriptive statistics. 
Changes from baseline in the SARA test scores will be summarized descriptively over time and by 
treatment cohort for each category and the total score. The SARA is a tool for assessing ataxia. It has 
eight categories with accumulative score ranging from 0 (no ataxia) to 40 (most severe ataxia). When 
completing the outcome measure, each category is assessed and scored accordingly. Scores for the eight 
items range as follows:  
1. Gait (0-8 points),  
2. Stance (0-6 points),  
3. Sitting (0-4 points),  
4. Speech disturbance (0-6 points),  
5. Finger chase (0-4 points),  
6. Nose-finger test (0-4 points),  
7. Fast alternating hand movement (0-4 points),  


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8. Heel-shin slide (0-4 points).  
Once each of the 8 categories has been assessed, the total is calculated to determine the severity of 
ataxia. 
For motor activities of the four extremities (Items 5-8), assessments are performed bilaterally, and the 
mean values are used to obtain the total score. 
8.4.4. 
Pharmacokinetic Analysis 
PK samples will be analyzed using a validated, sensitive, specific method. The laboratory analyzing the 
PK samples will be unblinded to ensure that PK analysis is performed only on subjects receiving 
RGLS8429. Placebo samples will be analyzed if necessary. Descriptive statistics for plasma 
concentrations by time point and by cohort will be tabulated. Descriptive statistics for PK parameters 
will be summarized by cohort.  
Concentration data will be presented and summarized for the PK Full population. 
Using non-compartmental methods, the plasma and urine concentration data will be used to derive the 
following PK parameters: Cmax, Tmax, AUC0-24, AUC0-t, AUCinf (where calculable), t½, CL/F, Vz/F, fe, 
and Ae. Additional PK parameters may be calculated as appropriate. The PK Evaluable Population will 
be used for these analyses. 
Dose proportionality will be explored using the power model for Cmax, AUC0-24, AUC0-t, and AUCinf as 
data permit. 
8.4.5. 
Biomarker Analysis 
Descriptive statistics will be used to characterize urine (e.g., PC1, PC2, NGAL, KIM-1, MCP-1, B2M, 
and CLU) and serum (e.g., Cystatin-C, IGFALS, Alb, CT-proAVP, N-acetyl-1-methylhistidine, and 
others) biomarkers in all subjects. The analysis of urine and serum biomarkers is exploratory. The PD 
Population will be used for these analyses. 


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9. 
ADVERSE EVENTS 
9.1. 
Monitoring, Recording and Reporting of Adverse Events 
An adverse event (AE) is any noxious, unintended, or untoward medical occurrence that may appear or 
worsen in a subject during a study. It may be a new intercurrent illness, a worsening concomitant illness, 
an injury, or any concomitant impairment of the subject’s health, including laboratory test values (as 
specified by the criteria in Section 9.3), regardless of etiology. Any worsening (i.e., any clinically 
significant adverse change in the frequency or intensity of a pre-existing condition) should be considered 
an AE. A diagnosis or syndrome should be recorded on the AE page of the CRF rather than the 
individual signs or symptoms of the diagnosis or syndrome.  
Overdose (accidental or intentional), abuse, withdrawal, sensitivity, or toxicity to a study drug should be 
reported as an AE. If an overdose is associated with an AE, the overdose and AE should be reported as 
separate terms. Any sequelae of an accidental or intentional overdose of a study drug should be reported 
as an AE on the AE CRF. If the sequelae of an overdose are a SAE, then the sequelae must be reported 
on the AE CRF and marked as serious. The overdose resulting in the SAE should be identified as the 
cause of the event on the CRF but should not be reported as an SAE itself.  
In the event of overdose, the subject should be monitored as appropriate and should receive supportive 
measures, as necessary. There is no known specific antidote for RGLS8429 overdose. Actual treatment 
should depend on the severity of the clinical situation and the judgment and experience of the treating 
physician. (Please refer to the RGLS8429 Investigator’s Brochure, Summary of Data and Guidance for 
the Investigator for additional information.) 
All subjects will be monitored for AEs during the study. Assessments may include monitoring of any or 
all the following parameters: the subject’s clinical symptoms, laboratory, pathological, radiological, or 
surgical findings, physical examination findings, or findings from other tests and/or procedures. 
All AEs and SAEs will be recorded by the Investigator with the start of study drug administration until 
EOS Visit. All SAEs made known to the Investigator at any time thereafter that are suspected of being 
related to study drug will also be recorded. AEs and SAEs will be recorded on the AE page of the CRF 
and in the subject’s source documents.  
9.2. 
Evaluation of Adverse Events 
A qualified Investigator must evaluate all adverse events.  
9.2.1. 
Seriousness  
An SAE is any AE occurring at any dose that: 
x Results in death. 
x Is life-threatening (i.e., in the opinion of the Investigator, the subject is at immediate risk of 
death from the AE). 
x Requires inpatient hospitalization or prolongation of existing hospitalization (hospitalization 
is defined as an inpatient admission, regardless of length of stay). 
x Results in persistent or significant disability/incapacity (a substantial disruption of the 
subject’s ability to conduct normal life functions). 
x Results in a congenital anomaly/birth defect. 


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x Constitutes an important medical event. Important medical events are defined as those 
occurrences that may not be immediately life-threatening or result in death, hospitalization, or 
disability, but may jeopardize the subject or require medical or surgical intervention to prevent one 
of the other outcomes listed above. Medical and scientific judgment should be exercised in 
deciding whether such an AE should be considered serious. 
Events not considered to be SAEs are hospitalizations for: 
x A standard procedure for protocol therapy administration. However, hospitalization or 
prolonged hospitalization for a complication of therapy administration will be reported as an 
SAE.  
x Routine treatment or monitoring of the studied indication not associated with any 
deterioration in condition. 
x The administration of blood or platelet transfusion as routine treatment of a concurrent 
medical condition. However, hospitalization or prolonged hospitalization for a complication 
of such transfusion remains a reportable SAE. 
x A procedure for protocol/disease-related investigations (e.g., surgery, scans, endoscopy, 
sampling for laboratory tests, bone marrow sampling). However, hospitalization or prolonged 
hospitalization for a complication of such procedures remains a reportable SAE. 
x Hospitalization or prolongation of hospitalization for technical, practical, or social reasons, in 
absence of an AE. 
x A procedure that is planned (i.e., planned prior to start of treatment on study); must be 
documented in the source document and the CRF. Hospitalization or prolonged 
hospitalization for a complication remains a reportable SAE. 
x An elective treatment of or an elective procedure for a pre-existing condition, unrelated to the 
studied indication that has not worsened from baseline.  
x Emergency outpatient treatment or observation that does not result in admission, unless 
fulfilling other seriousness criteria above. 
If an AE is considered serious, the AE page/screen of the CRF must be completed and marked as 
serious. 
For each SAE, the Investigator will provide information on severity, start, and stop dates, relationship to 
the study drug, action taken regarding the study drug, and outcome. 
9.2.2. 
Severity/Intensity 
For both AEs and SAEs, the Investigator must assess the severity/intensity of the event.  
The Investigator will assess the grade of the AE according to the FDA’s Guidance for Industry: Toxicity 
Grading Scale for Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical 
Trials (September 2007) except for AEs associated with abnormal serum creatinine level. The 
Investigator will use the KDIGO Clinical Practice Guideline for Acute Kidney Injury 
(https://kdigo.org/wp-content/uploads/2016/10/KDIGO-2012-AKI-Guideline-English.pdf) to grade the 
severity of an AE associated with abnormal serum creatinine level. Toxicities that are not specified in 
these guidances will be defined as follows:  


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x Grade 1: Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; 
intervention not indicated 
x Grade 2: Moderate; minimal, local, or noninvasive intervention indicated; limiting age-
appropriate instrumental activities of daily living (ADL) 
x Grade 3: Severe or medically significant but not immediately life-threatening; hospitalization 
or prolongation of hospitalization indicated; disabling; limiting self-care ADL 
x Grade 4: Life-threatening consequences; urgent intervention indicated 
x Grade 5: Death related to AE 
The term “severe” is often used to describe the intensity of a specific event (as in mild, moderate, or 
severe myocardial infarction); however, the event itself may be of relatively minor medical significance 
(such as severe headache). This criterion is not the same as “serious”, which is based on subject/event 
outcome or action criteria associated with events that pose a threat to a subject’s life or functioning. 
Seriousness, not severity, serves as a guide for defining regulatory obligations. 
9.2.3. 
Causality 
The Investigator must determine the relationship between the study drug and the occurrence of an 
AE/SAE. This relationship is indicated by as “unrelated” or “related” in response to the question: Is 
there a reasonable possibility that the event may be caused by the study drug. In addition to a temporal 
relationship, the Investigator should consider the following when determining the relationship between 
an AE and study drug: whether an alternative etiology has been identified, mechanism of action of the 
study drug, and/or the biological plausibility of a relationship. 
9.2.4. 
Duration 
For both AEs and SAEs, the Investigator will provide a record of the start and stop dates of the event. In 
all cases, the AE must have the same verbatim term throughout. Within the duration of the SAE or AE, 
the maximum grade should be used to categorize severity. 
9.2.5. 
Action Taken 
The Investigator will report the action taken with study drug as a result of an AE or SAE, as applicable 
and report if concomitant and/or additional treatments were given for the event.  
9.2.6. 
Outcome 
Subjects will be followed until AEs have either resolved, returned to baseline status, or are deemed 
stable or commensurate with ongoing disease processes, per the Investigator’s judgment.  
One of the four outcomes listed below must be recorded:  
x Resolved – The subject has recovered fully from the event with no residual effects 
observable or returned to baseline status.  
x Resolved with sequelae – The subject has recovered from the event with some residual 
effect(s) observable. The residual effect(s) will be recorded in the CRF.  


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x Ongoing – Effects of the event are still present, regardless of whether the effect is changing 
or stable and persistent.  
x Fatal outcome (for serious adverse events only)  
9.3. 
Abnormal Laboratory Values 
An abnormal laboratory value is an AE if the abnormality: 
x Results in discontinuation from the study. 
x Requires treatment, stopping or holding study drug, or any other therapeutic intervention. 
x Is judged to be clinically significant, e.g., one that indicates a new disease process and/or 
organ toxicity; or is an exacerbation or worsening of an existing condition. 
Regardless of severity grade, only laboratory abnormalities that fulfill a seriousness criterion need to be 
documented as a serious adverse event. 
If a laboratory abnormality is one component of a diagnosis or syndrome, then only the diagnosis or 
syndrome should be recorded on the AE page/screen of the CRF. If the abnormality was not a part of a 
diagnosis or syndrome, then the laboratory abnormality should be recorded as the AE. If possible, the 
laboratory abnormality should be recorded as a medical term and not simply as an abnormal laboratory 
result (e.g., record thrombocytopenia rather than decreased platelets). 
9.4. 
Pregnancy 
All pregnancies or suspected pregnancies occurring in either a female subject or female partner of a 
male subject are immediately reportable events.  
In the event that a pregnancy or suspHFWHGSUHJQDQF\LQFOXGLQJHOHYDWHGȕK&*RUSRVLWLYHSUHJQDQF\
test in a female subject) should occur while the subject is in study, or within 90 days of the subject’s 
dose of study drug, it will be considered an immediately reportable event. The pregnancy, suspected 
pregnancy, or positive pregnancy test must be reported to the study pharmacovigilance group 
immediately by email using the Pregnancy Reporting Form. 
The female subject may be referred to an obstetrician-gynecologist or another appropriate healthcare 
professional for further evaluation. 
The Investigator will follow the female subject until completion of the pregnancy and must notify the 
study pharmacovigilance group immediately about the outcome of the pregnancy (normal or abnormal 
outcome) using the Pregnancy Follow-up Report Form. 
If the outcome of the pregnancy was abnormal (e.g., spontaneous abortion), the Investigator should 
report the abnormal outcome as an AE. If the abnormal outcome meets any of the serious criteria, it 
must be reported as an SAE to the study pharmacovigilance group within 24 hr of the Investigator’s 
knowledge of the event. 
All neonatal deaths that occur within 28 days of birth should be reported, without regard to causality, as 
SAEs. In addition, any infant death after 28 days that the Investigator suspects is related to the in-utero 
exposure to the study drug should also be reported to the study pharmacovigilance group within 24 hr of 
the Investigator’s knowledge of the event. 


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9.4.1. 
Male Subjects 
If a female partner of a male subject taking study drug becomes pregnant, the male subject taking study 
drug should notify the Investigator, and the pregnant female partner should be advised to call their 
healthcare provider immediately.  
9.5. 
Reporting of Serious Adverse Events 
All AEs that meet criterion for an SAE must be entered on the AE CRF and marked as serious in the 
clinical database. All SAEs must be reported to the study pharmacovigilance group within 24 hr of 
knowledge of the event.  
The Investigator is required to ensure that the data on these forms is accurate and consistent. This 
requirement applies to all SAEs (regardless of relationship to study drug) that occur during the study 
(from the administration of study drug until the EOS Visit) or any SAE made known to the Investigator 
at any time thereafter that are suspected of being related to study drug.  
The SAE report should provide a detailed description of the SAE and include a concise summary of 
hospital records and other relevant documents. If a subject died and an autopsy has been performed, copies 
of the autopsy report and death certificate are to be sent to the study PV mailbox as soon as these become 
available. Any follow-up data should be detailed in the AE CRF.  
Where required by local legislation, the Investigator is responsible for informing the Institutional 
Review Board/Ethics Committee (IRB/EC) of the SAE and providing them with all relevant initial and 
follow-up information about the event. The Investigator must keep copies of all SAE information on file 
including correspondence with the study pharmacovigilance group and the IRB/EC. 
9.6. 
Safety Queries 
Queries pertaining to SAEs will be communicated from the study pharmacovigilance group to the site 
via electronic mail. The response time is expected to be no more than five (5) business days. Urgent 
queries (e.g., missing causality assessment) may be handled by phone. 
9.7. 
Expedited Reporting of Adverse Events 
For the purpose of regulatory reporting, the Sponsor’s Lead Physician will determine the expectedness 
of events suspected of being related to study drug based on information in the Reference Safety 
Information section of the Investigator Brochure.  
All suspected unexpected serious adverse reactions (SUSARs) will be reported in an expedited manner 
in accordance local and FDA (21 CFR 312.32) regulations. 
The study pharmacovigilance group shall notify the Investigator of the following information: 
x Any AE suspected of being related to the use of study drug in this study or in other studies 
that is both serious and unexpected (i.e., SUSAR). 
x Any finding from tests in laboratory animals that suggests a significant risk for human 
subjects including reports of mutagenicity, teratogenicity, or carcinogenicity. 
Where required by local legislation, the Investigator shall notify his/her IRB/EC promptly of these new 
serious and unexpected AE(s) or significant risks to subjects. 


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The Investigator must keep copies of all pertinent safety information on file including correspondence 
with Regulus and the IRB/EC. (See Section 13.3 for record retention information). 
9.8. 
Study Pharmacovigilance Group Contact Information 
See Study Reference Manual 


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10. 
DISCONTINUATIONS 
The following events are considered valid reasons for discontinuing a subject from the study drug and/or 
from the study: 
x Protocol violation 
x Non-compliance 
x Adverse event 
x Withdrawal by subject 
x Subject lost to follow-up 
x Sponsor decision 
x Investigator decision 
x Pregnancy or pregnancy of partner 
x Death 
x Other  
The reason for discontinuation should be recorded in the CRF and in the source documents. 
The decision to discontinue a subject remains the responsibility of the treating physician, which will not 
be delayed or refused by the Sponsor. However, prior to discontinuing a subject, the Investigator should 
(if possible) contact the Sponsor’s Lead Physician and forward appropriate supporting documents (if 
possible) for review and discussion. 


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11. 
EMERGENCY PROCEDURES 
11.1. 
Emergency Contact 
In emergency situations, the Investigator should provide for appropriate emergency care without delay 
and contact the Sponsor’s Lead Physician or designee by telephone at the number(s) listed in the Study 
Reference Manual. 


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12. 
REGULATORY CONSIDERATIONS 
12.1. 
Good Clinical Practice 
The procedures set out in this study protocol pertaining to the conduct, evaluation, and documentation of 
this study are designed to ensure that the Sponsor, its authorized representative, and Investigator abide 
by Good Clinical Practice (GCP), as described in International Conference on Harmonisation (ICH) 
Guideline E6 (R2) and in accordance with the general ethical principles outlined in the Declaration of 
Helsinki. The study will receive approval from an IRB/EC prior to commencement. The Investigator 
will conduct all aspects of this study in accordance with applicable national, state, and local laws of the 
pertinent regulatory authorities. 
12.2. 
Investigator Responsibilities 
Investigator responsibilities are set out in the ICH Guideline for Good Clinical Practice E6 (R2) and in 
the local regulations. Regulus staff or an authorized representative will evaluate and approve all 
Investigators who in turn will select their staff. 
The Investigator should ensure that all persons assisting with the study are adequately informed about 
the protocol, amendments, study treatments, as well as study-related duties and functions, including 
obligations of confidentiality of Regulus information. The Investigator should maintain a list of Sub-
Investigators and other appropriately qualified persons to whom he or she has delegated significant 
study-related duties. 
The Investigator is responsible for keeping a record of all subjects who sign an informed consent form 
(ICF) and are screened for entry into the study. Subjects who fail Screening must have the reason(s) 
recorded in the subject’s source documents. 
The Investigator, or a designated member of the Investigator’s staff, must be available during 
monitoring visits to review data, resolve queries and allow direct access to subject records (e.g., medical 
records, office charts, hospital charts, and study-related charts) for source data verification. The 
Investigator must ensure timely and accurate completion of CRFs and queries. 
The information contained in the protocol and amendments (except for the information provided by 
Regulus on public registry websites) is considered Regulus confidential information. Only information 
that is disclosed previously by Regulus on a public registry website may be freely disclosed by the 
Investigator or its institution, or as outlined in the Clinical Trial Agreement. The Regulus protocol, 
amendments, and IB information are not to be made publicly available (for example on the 
Investigator’s or their institution’s website) without express written approval from Regulus. Information 
proposed for posting on the Investigator’s or their institution’s website must be submitted to Regulus for 
review and approval, providing at least 5 business days for review. 
At the time results of this study are made available to the public, if requested, Regulus will provide 
Investigators with a summary of the results that is written for the lay person. The Investigator is 
responsible for sharing these results with the subject and/or their caregiver as agreed by the subject. 
12.3. 
Subject Information and Informed Consent 
The Investigator must obtain informed consent of a subject and/or a subject’s legal representative prior 
to any study-related procedures. 


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Documentation that informed consent occurred prior to the study subject’s entry into the study and of 
the informed consent process should be recorded in the study subject’s source documents including the 
date. The original ICF signed and dated by the study subject and by the person consenting the study 
subject prior to the study subject’s entry into the study, must be maintained in the Investigator’s study 
files and a copy given to the study subject. In addition, if a protocol is amended and it impacts on the 
content of the informed consent, the ICF must be revised. Study subjects participating in the study when 
the amended protocol is implemented must be re-consented with the revised version of the ICF. The 
revised ICF signed and dated by the study subject and by the person consenting the study subject must 
be maintained in the Investigator’s study files and a copy given to the study subject. 
12.4. 
Confidentiality 
Regulus affirms the subject's right to protection against invasion of privacy and to follow ICH and other 
local regulations (whichever is most stringent). Regulus requires the Investigator to permit Regulus 
representatives and, when necessary, representatives from regulatory authorities, to review and/or copy 
any medical records relevant to the study in accordance with local laws. 
Should direct access to medical records require a waiver or authorization separate from the subject’s 
signed ICF, it is the responsibility of the Investigator to obtain such permission in writing from the 
appropriate individual. 
12.5. 
Protocol Amendments 
Any amendment to this protocol must be approved by the Sponsor and submitted to the IRB/EC for 
written approval. Written approval must be obtained before implementation of the amended version 
occurs. The written signed approval from the IRB/EC should specifically reference the Investigator 
name, protocol number, study title, and amendment number(s) that are applicable. Amendments that are 
administrative in nature do not require IRB/IEC approval but will be submitted to the IRB/IEC for 
information purposes. 
12.6. 
Institutional Review Board/Independent Ethics Committee Review and Approval 
Before the start of the study, the study protocol, ICF, and any other appropriate documents will be 
submitted to the IRB/EC with a cover letter or a form listing the documents submitted, their dates of 
issue, and the site (or region or area of jurisdiction, as applicable) for which approval is sought. If 
applicable, the documents will also be submitted to the authorities in accordance with local legal 
requirements. 
RGLS8429 can be supplied to an Investigator by Regulus or its authorized representative only after 
documentation on all ethical and legal requirements for starting the study has been received by Regulus 
or its authorized representative. This documentation must also include a list of the members of the 
IRB/EC and their occupation and qualifications. If the IRB/EC will not disclose the names, occupations, 
and qualifications of the committee members, it should be asked to issue a statement confirming that the 
composition of the committee is in accordance with GCP. For example, the IRB General Assurance 
Number may be accepted as a substitute for this list. Formal approval by the IRB/EC should mention the 
protocol title, number, amendment number (if applicable), study site (or region or area of jurisdiction, as 
applicable), and any other documents reviewed. It must mention the date on which the decision was 
made and must be officially signed by a committee member. Before the first subject is enrolled in the 
study, all ethical and legal requirements must be met. 


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The IRB/EC and, if applicable, the authorities, must be informed of all subsequent protocol amendments 
in accordance with local legal requirements. Amendments must be evaluated to determine whether 
formal approval must be sought and whether the ICF should also be revised. 
The Investigator must keep a record of all communication with the IRB/EC and, if applicable, between a 
Coordinating Investigator and the IRB/EC. This statement also applies to any communication between 
the Investigator (or Coordinating Investigator, if applicable) and regulatory authorities. 
Any advertisements used to recruit subjects for the study must be reviewed by Regulus and the IRB/EC 
prior to use.  
12.7. 
Ongoing Information for Institutional Review Board/ Ethics Committee 
If required by legislation or the IRB/EC, the Investigator must submit to the IRB/EC: 
x Information on serious or unexpected adverse events as soon as possible  
x Periodic reports on the progress of the study 
x Deviations from the protocol or anything that may involve added risk to subjects in 
accordance with IRB/EC guidelines 
12.8. 
Termination of the Study 
Regulus reserves the right to terminate this study at any time for reasonable medical or administrative 
reasons. Any premature discontinuation will be documented appropriately according to local 
requirements (e.g., IRB/EC, regulatory authorities, etc.). 
In addition, the Investigator or Regulus has the right to discontinue a single site at any time during the 
study for medical or administrative reasons such as: 
x Unsatisfactory enrollment 
x GCP noncompliance 
x Inaccurate or incomplete data collection 
x Falsification of records 
x Failure to adhere to the study protocol 


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13. 
DATA HANDLING AND RECORDKEEPING  
13.1. 
Data/Documents 
The Investigator must ensure that the records and documents pertaining to the conduct of the study and 
the distribution of the study drug are complete, accurate, filed, and retained. Examples of source 
documents include hospital records; clinic and office charts; laboratory notes; memoranda; subject’s 
diaries or evaluation checklists; dispensing records; recorded data from automated instruments; copies or 
transcriptions certified after verification as being accurate copies; microfiche; x-ray film and reports; 
and records kept at the pharmacy, and the laboratories, as well as copies of CRFs. 
13.2. 
Data Management 
Data will be collected via CRF, and data entry will be performed through username- and password-
protected access to a secure database. These data will be verified electronically through use of 
programmed edit checks specified by the Sponsor. Discrepancies in the data will be brought to the 
attention of the Sponsor and investigational site personnel, as necessary. Resolutions to these issues will 
be reflected in the database. An audit trail within the system will track all changes made to the data. 
13.3. 
Record Retention 
Essential documents must be retained by the Investigator according to the period outlined in the Clinical 
Trial Agreement. The Investigator must retain these documents for the time period described above or 
according to local laws or requirements, whichever is longer. Essential documents include, but are not 
limited to, the following: 
x Signed ICFs for all subjects 
x Subject identification code list, Screening log (if applicable), and enrollment log 
x Record of all communications between the Investigator and the IRB/EC 
x Composition of the IRB/EC 
x Record of all communications between the Investigator, Regulus, and their authorized 
representative(s) 
x List of Sub-Investigators and other appropriately qualified persons to whom the Investigator 
has delegated significant study-related duties, together with their roles in the study, 
curriculum vitae, and their signatures 
x RGLS8429 accountability records 
x Record of any body fluids or tissue samples retained 
x All other source documents (subject records, hospital records, laboratory records, etc.) 
x All other documents as listed in Section 8 of the ICH consolidated guideline on GCP 
(Essential Documents for the Conduct of a Clinical Trial) 
The Investigator must notify Regulus if he/she wishes to assign the essential documents to someone else, 
remove them to another location, or is unable to retain them for a specified period. The Investigator 
must obtain approval in writing from Regulus prior to destruction of any records. If the Investigator is 


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unable to meet this obligation, the Investigator must ask Regulus for permission to make alternative 
arrangements. Details of these arrangements should be documented.  
All study documents should be made available if required by relevant health authorities. Investigator or 
institution should take measures to prevent accidental or premature destruction of these documents. 


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14. 
QUALITY CONTROL AND QUALITY ASSURANCE 
All aspects of the study will be monitored carefully by Regulus or its authorized representative for 
compliance with applicable government regulations with respect to current GCP and SOPs. 
14.1. 
Study Monitoring and Source Data Verification 
Regulus or designee ensures that appropriate monitoring procedures are performed before, during, and 
after the study. All aspects of the study are reviewed with the Investigator and the staff at a study 
initiation visit and/or at an Investigators’ Meeting. Prior to enrolling subjects into the study, a Regulus 
representative or designee will review the protocol, CRFs, procedures for obtaining informed consent, 
record keeping, and reporting of AEs/SAEs with the Investigator. Monitoring will include on-site visits 
with the Investigator and his/her staff as well as any appropriate communications by mail, email, or 
telephone. During monitoring visits, the facilities, study drug storage area, CRFs, subject’s source 
documents, and all other study documentation will be inspected/reviewed by the Regulus representative. 
Accuracy will be checked by performing source data verification that is a direct comparison of the 
entries made onto the CRFs against the appropriate source documentation. Any resulting discrepancies 
will be reviewed with the Investigator and/or his/her staff. Any necessary corrections will be made 
directly to the CRFs or via queries by the Investigator and/or his/her staff. Monitoring procedures 
require that informed consents, adherence to inclusion/exclusion criteria, and documentation of SAEs 
and their proper recording be verified. Additional monitoring activities may be outlined in a study-
specific monitoring plan. 
14.2. 
Audits and Inspections 
In addition to the routine monitoring procedures, a representative of Regulus or designee may conduct 
audits of clinical research activities to evaluate compliance with Good Clinical Practice guidelines and 
regulations. 
The Investigator is required to permit direct access to the facilities where the study took place, source 
documents, CRFs, and applicable supporting records of study subject participation for audits and 
inspections by IRB/ECs, regulatory authorities, and company-authorized representatives. The 
Investigator should make every effort to be available for the audits and/or inspections. If the Investigator 
is contacted by any regulatory authority regarding an inspection, he/she should contact Regulus or 
designee immediately. 


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15. 
PUBLICATIONS 
As described in Section 12.2, all protocol- and amendment-related information, with the exception of the 
information provided by Regulus or designee on public registry websites, is considered confidential 
information and is not to be used in any publications without prior approval. Regulus protocol-related 
information proposed for use in a publication must be submitted to Regulus for review and approval and 
should not be utilized in a publication without express written approval from Regulus, or as described in 
the Clinical Trial Agreement. 
Regulus will ensure Regulus-sponsored studies are considered for publication in the scientific literature 
in a peer-reviewed journal, irrespective of the results. At a minimum, this applies to results from clinical 
studies, and any other study results of significant medical importance. 
Study results may also be presented at one or more medical congresses and may be used for scientific 
exchange and teaching purposes. Additionally, this study and its results may be submitted for inclusion 
in all appropriate health authority study registries, as well as publication on health authority study 
registry websites, as required by local health authority regulations. 
Eligibility for external authorship, as well as selection of first authorship, will be based on several 
considerations, including, but not limited to, contribution to protocol development, study recruitment, 
data quality, participation in data analysis, participation in study steering committee (when applicable), 
and contribution to abstract, presentation, and/or publication development. 


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16. 
REFERENCES  
Chapman AB, Devuyst O, Eckardt DU, et al. Autosomal-dominant polycystic kidney disease 
(ADPKD): executive summary from a Kidney Disease: Improving Global Outcomes (KDIGO) 
Controversies Conference. Kidney Int. 2015;88(1):17-27. 
Chebib FT, Torres VE. Autosomal Dominant Polycystic Kidney Disease: Core Curriculum 2016. 
Am J Kidney Dis. 2016;67(5):792-810.  
Dong K, Zhang, C, Tian X, Coman, D, Hyder F, Ma M, Somlo, S. Renal plasticity revealed through 
reversal of polycystic kidney disease in mice. Nature Genetics. 2021;1-15. 10.1038/s41588-021-
00946-4. 
Hajarnis S, Lakhia R, Yheskel M, Williams D, Sorourian M, Liu X, Aboudehen K, Zhang S, Kersjes 
K, Galasso R, Li J, Kaimal V, Lockton S, Davis S, Flaten A, Johnson JA, Holland WL, Kusminski 
CM, Scherer PE, Harris PC, Trudel M, Wallace DP, Igarashi P, Lee EC, Androsavich JR, Patel V. 
microRNA-17 family promotes polycystic kidney disease progression through modulation of 
mitochondrial metabolism. Nat Commun. 2017 Feb 16;8:14395. 
Hogan MC, Bakeberg JL, Gainullin VG, Irazabal MV, Harmon AJ, Lieske JC, Charlesworth MC, 
Johnson KL, Madden BJ, Zenka RM, McCormick DJ, Sundsbak JL, Heyer CM, Torres VE, Harris 
PC, Ward CJ. Identification of Biomarkers for PKD1 Using Urinary Exosomes. J Am Soc Nephrol. 
2015 Jul;26(7):1661-70.  
Inker LA, Eneanya ND, Coresh J, et al. New creatinine- and cystatin C–based equations to estimate 
GFR without race. N Engl J Med 2021;385:1737-49. DOI: 10.1056/NEJMoa2102953 
Lee E,Valencia T, Allerson C, Schairer A, Flaten A, Yheskel, M, Kersjes K, Li J, Gatto, S, Takhar 
M, Lockton S, Pavlicek A, Kim M, Chu T, Soriano R, Davis S, Androsavich J, Sarwary S, Owen T, 
Patel V. Discovery and preclinical evaluation of anti-miR-17 oligonucleotide RGLS4326 for the 
treatment of polycystic kidney disease. Nature Communications. 2019;10. 1-14. 10. 
Lee E, Valencia TM, Owen T, Neben S, Cremer KF, Garg R, Drygin D, Ward CJ, Yu ASL, Patel V. 
RGLS4326 Increases Urinary PC1 and PC2 Levels in Individuals with Autosomal Dominant 
Polycystic Kidney Disease (ADPKD). American Society of Nephrology. Abstract PO1244. 2021 
Patel V, Williams D, Hajarnis S, Hunter R, Pontoglio M, Somlo S, Igarashi P. miR-17~92 miRNA 
cluster promotes kidney cyst growth in polycystic kidney disease. Proc Natl Acad Sci U S A. 2013 
Jun 25;110(26):10765-70.  
Torres VE, Harris PC. Autosomal dominant polycystic kidney disease: the last 3 years. Kidney Int. 
2009;76:149-168. 
 
 


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APPENDIX A. 
TABLE OF ABBREVIATIONS 
Abbreviation  
Explanation 
%CV 
Coefficient of variation 
ADA 
Anti-drug antibodies 
ADPKD 
Autosomal dominant polycystic kidney disease 
ADL 
Activities of daily living 
Ae 
Total amount of unchanged RGLS8429 excreted in the urine 
AE 
Adverse event 
Alb 
Albumin 
ALT 
Alanine aminotransferase  
AUC0-24 
Area under the concentration–time curve up to 24 h post dose 
AUC0-t 
Area under the concentration–time curve up to the last quantifiable concentration 
AUCinf 
Area under the concentration–time curve extrapolated to infinity 
aPTT 
Activated partial thromboplastin time 
AST 
Aspartate aminotransferase  
BMI  
Body mass index  
B2M 
beta-2 microglobulin 
BP 
Blood pressure 
BUN  
Blood urea nitrogen  
CKD-EPI 
Chronic kidney disease epidemiology collaboration equation 
CL/F 
Apparent clearance 
Cmax 
Maximum observed concentration 
CT-proAVP 
 
CK 
Creatine kinase 
CRF 
Case report form 
CRO 
Clinical Research Organization 
DLT 
Dose-limiting toxicity 
ECG  
Electrocardiogram  
EOS  
End of study  
fe 
Fraction of unchanged RGLS8429 excreted in the urine 
GCP  
Good clinical practice  


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Abbreviation  
Explanation 
GGT  
Gamma-glutamyl Transferase 
GLP 
Good Laboratory Practice GLP 
Hb 
Hemoglobin 
Hct 
Hematocrit 
HED 
Human equivalent dose 
HIV 
Human immunodeficiency virus 
HR 
Heart rate 
IGFALS 
Insulin-like Growth Factor Binding Protein Acid Labile Subunit  
ICF 
Informed consent form  
ICH  
International Conference on Harmonisation  
INR  
International normalized ratio  
IUD 
Intrauterine device 
IUS 
Intrauterine system 
KIM-1 
Kidney injury molecule 1 
LDH  
Lactate dehydrogenase  
MCH 
Mean corpuscular hemoglobin 
MCHC 
Mean corpuscular hemoglobin concentration 
MCP-1 
monocyte chemoattractant protein-1 (MCP-1) 
MCV 
Mean corpuscular volume 
miRNA 
MicroRNA 
MTD 
Maximum tolerated dose 
ncRNA 
Non-coding microRNA 
mmHg 
Millimeters of mercury  
MRI  
Magnetic resonance imaging  
NOAEL 
No-observed-adverse-effect Level 
NGAL 
Neutrophil gelatinase-associated lipocalin 
PC1 
Polycystin 1 
PC2 
Polycystin 2 
PD 
Pharmacodynamic 
PK 
Pharmacokinetic 


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Abbreviation  
Explanation 
PKD 
Polycystic kidney disease 
PT  
Prothrombin time  
Q2W 
Every other week 
RBC  
Red blood cells  
RR 
Respiratory rate 
SARA 
Scale for the Assessment and Rating of Ataxia 
SAE  
Serious adverse event  
SC 
Subcutaneous 
SOP  
Standard operating procedure  
SUSAR  
Suspected unexpected serious adverse reaction  
Tmax 
Time to maximum observed concentration 
TEAE  
Treatment emergent adverse event  
t½ 
Half-life 
ULN  
Upper limit of normal  
Vz/F 
Volume of distribution 
WBC  
White blood cells  


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APPENDIX B. 
SCALE FOR THE ASSESSMENT AND RATING OF ATAXIA 
(SARA) 
Schmitz-Hübsch T, Tezenes du Montcel S, Baliko L, et al: Scale for the assessment and rating of ataxia: 
development of a new clinical scale. Neurology 66:1717-1720, 2006. 
 


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SAP_001.pdf: 
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Protocol. No. RGLS8429-01
ICON Project No. 3877/0009 
Statistical Analysis Plan Final v1.0
19 Sep 2022 
CONFIDENTIAL
Page 1 of 32
STATISTICAL ANALYSIS PLAN
A Phase 1, Double-Blind, Placebo-Controlled, Single Ascending Dose Study in 
Healthy Volunteers to Evaluate the Safety, Tolerability, and 
Pharmacokinetics of RGLS8429
SAP Version 1.0
Final
Date: 19 Sep 2022
for
Protocol No. RGLS8429-01
Submitted to:
Regulus Therapeutics Inc.
4224 Campus Point Court
Suite 210
San Diego, CA 92121
United States
Prepared by:
ICON Clinical Research, LLC
820 West Diamond Avenue, Suite 100
Gaithersburg, MD 20878
Telephone: (301) 944-6800
Fax: (215) 699-6288
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SIGNATURES
ICON Early Phase Services, LLC Signature Page
ICON Author: 
Siva Durga Prasad Sattu, M.Sc.
Senior Statistician II, Biostatistics
Date
ICON Reviewers:
 
Shekar Sunkoju 
Senior Manager, Biostatistics
Date
Rakesh Govindaraj
Principal SAS Programmer, IEP SAS Programming
Date
Laurie Reynolds.
Sr. PK Scientist, Quantitative Pharmacology & 
Pharmacometrics
Date
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d4744496-6a55-4b4f-98f6-4abf57c4707c
Shekar Sunkoju
19 Sep 2022 03:01:15 PDT (-07:00)
REASON: I approve this document
d46665cd-bd3c-4abf-9de6-c3fbe2e057ae
Siva Durga Prasad Sattu
19 Sep 2022 21:14:35 PDT (-07:00)
REASON: I approve this document as author.
35960b85-4e6f-4d93-9975-f7e48313876f
Rakesh Govindaraj
20 Sep 2022 04:26:34 PDT (-07:00)
REASON: I approve this document
d6ad1f2c-7b30-4b99-ad94-1e44ce82e6be
Laurie Reynolds
21 Sep 2022 06:44:02 PDT (-07:00)
REASON: I approve this document


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CLIENT SIGNATURE PAGE
Client Approvals:
Rekha Garg, MD, MS
Clinical Development and Regulatory Consultant
Date
Claire S. Padgett, PhD
Clinical Consultant 
Date
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4b0a0d9e-bf5c-4fae-b106-69a797d0235a
Claire Padgett
21 Sep 2022 09:07:20 PDT (-07:00)
REASON: I approve this document
c465b3ea-142a-4c12-a06c-8863f5f6a2ed
Rekha Garg
23 Sep 2022 07:06:37 PDT (-07:00)
REASON: I approve this document


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LIST OF ABBREVIATIONS
Abbreviation Explanation
Ae
Total amount of unchanged RGLS8429 excreted in the urine
AE
Adverse event
Alb
Albumin
ALP
Alkaline phosphatase
ALT
Alanine aminotransferase
aPTT
Activated partial thromboplastin time
AST
Aspartate aminotransferase
AUC
Area under the curve
AUC0-24
Area under the concentration–time curve up to 24 hr post dose
AUC0-t
Area under the concentration–time curve up to the last quantifiable 
concentration
AUCinf
Area under the concentration–time curve extrapolated to infinity
BLQ
Below limit of quantitation
BMI
Body mass index
B2M
beta-2 microglobulin
BP
Blood pressure
BUN
Blood Urea Nitrogen
CL/F
Apparent clearance
CLR
Renal Clearance
CLU
Clusterin 
cm
centimetre
Cmax
Maximum observed concentration
CT-proAVP
Copeptin
CRA
Clinical research associate
CRF
Case report form
CRO
Clinical research organization
CS
Clinically significant
CSR
Clinical study report
CV%
Coefficient of variation
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Abbreviation Explanation
DMP
Data Management Plan
DLT
Dose-limiting toxicity
ECG
Electrocardiogram
eCRF
Electronic case report form
eGFR
Estimated glomerular filtration rate
EOS
End of study
EPQT
Early Precision QT
Fe
Fraction of unchanged RGLS8429 excreted in the urine
GCP
Good clinical practice
GGT
Gamma-glutamyl transferase
Hb
Hemoglobin
Hct
Hematocrit
HIV
Human immunodeficiency virus
hr
Hour
HR
Heart rate
ICH
International Conference on Harmonisation
IGFALS
Insulin-like Growth Factor Binding Protein Acid Labile Subunit
INR
International normalized ratio
kg
Kilogram
KIM-1
Kidney injury molecule 1
LDH
Lactate dehydrogenase
MCH
Mean corpuscular hemoglobin
MCHC
Mean corpuscular hemoglobin concentration
MCP-1
Monocyte chemoattractant protein-1 (MCP-1)
MCV
Mean corpuscular volume
MedDRA
Medical dictionary for regulatory activities
MTD
Maximum tolerated dose
NGAL
Neutrophil gelatinase-associated lipocalin
OTC
Over-the-counter
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Abbreviation Explanation
PC1
Polycystin 1
PC2 
Polycystin 2
PD
Pharmacodynamic(s)
PK
Pharmacokinetic(s)
PR (interval)
Interval measured from the beginning of the P wave to the beginning of the
QRS complex in the heart’s electrical cycle as measured by electrocardiogram
PT
Preferred Term
QRS
The interval between the Q wave and the S wave in the heart’s electrical cycle 
as measured by electrocardiogram; principal deflection in the 
electrocardiogram
QT
Measure of time between the start of the Q wave and the end of the T wave in 
the heart’s electrical cycle
QTcB
QT correction with Bazett formula
QTcF
QT interval corrected by Fridericia
Q1
First Quartile
Q3
Third Quartile
RBC 
Red blood cells
RR
Respiratory rate
SAD
Single Ascending Dose
SAE 
Serious adverse event
SAP
Statistical analysis plan
SARA
Scale for the Assessment and Rating of Ataxia
SC
Subcutaneous
SD
Standard deviation
SOC
System Organ Class
t½
Terminal half-life
TEAE
Treatment emergent adverse event
Tmax
Time to maximum observed concentration
Vz/F
Apparent Volume of distribution
WBC
White blood cells
WHO
World Health Organization
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TABLE OF CONTENTS
SIGNATURES................................................................................................................................ 2 
CLIENT SIGNATURE PAGE ....................................................................................................... 3 
LIST OF ABBREVIATIONS......................................................................................................... 4 
TABLE OF CONTENTS................................................................................................................ 7 
1. 
INTRODUCTION................................................................................................................... 9 
2. 
STUDY OBJECTIVES AND ENDPOINTS ........................................................................ 10 
2.1 
Objectives ................................................................................................................... 10 
2.1.1 
Primary Objective ................................................................................................... 10 
2.1.2 
Secondary Objectives.............................................................................................. 10 
2.1.3 
Exploratory Objective............................................................................................. 10 
2.2 
Endpoints.................................................................................................................... 10 
2.2.1 
Primary Endpoint.................................................................................................... 10 
2.2.2 
Secondary Endpoints .............................................................................................. 10 
2.2.3 
Exploratory Endpoints ............................................................................................ 11 
3 
STUDY DESIGN .................................................................................................................. 12 
3.1 
General........................................................................................................................ 12 
3.2 
Study Population......................................................................................................... 15 
3.3 
Randomization and Treatment Assignments.............................................................. 15 
3.4 
Determination of Sample Size.................................................................................... 15 
4 
CHANGES IN THE CONDUCT OF THE STUDY OR PLANNED ANALYSIS.............. 16 
5 
QUALITY CONTROL AND QUALITY ASSURANCE METHODS FOR DATA 
ANALYSIS................................................................................................................................... 17 
6 
PHARMACOKINETIC AND PHARMACODYNAMIC ASSESSMENTS ....................... 18 
6.1 
Pharmacokinetic Assessment ..................................................................................... 18 
6.1.1 
Treatment of Outliers.............................................................................................. 19 
6.1.2 
Non-Quantifiable Concentrations........................................................................... 19 
6.2 
Biomarker Assessment ............................................................................................... 19 
7 
STATISTICAL METHODS.................................................................................................. 20 
7.1 
General........................................................................................................................ 20 
7.2 
Handling of Dropouts or Missing Data ...................................................................... 20 
7.2.1 
Handling of missing/ incomplete dates for Adverse Event .................................... 21 
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7.2.2 
Handling missing or partial Prior/Concomitant Medication Dates......................... 21 
7.3 
Multiple Comparisons and Multiplicity ..................................................................... 21 
7.4 
Adjustments for covariates ......................................................................................... 21 
7.5 
Multicenter Studies..................................................................................................... 22 
7.6 
Examination of Subgroups ......................................................................................... 22 
7.7 
Coding dictionaries..................................................................................................... 22 
7.8 
Analysis Populations .................................................................................................. 22 
7.8.1 
Screened Population................................................................................................ 22 
7.8.2 
Randomized Population.......................................................................................... 22 
7.8.3 
Safety Population.................................................................................................... 22 
7.8.4 
Pharmacokinetic Full Population............................................................................ 22 
7.8.5 
Pharmacokinetic Evaluable Population .................................................................. 22 
7.8.6 
Biomarkers (Pharmacodynamic (PD)) Population ................................................. 22 
7.9 
Subject Accountability ............................................................................................... 23 
7.10 Protocol Deviations .................................................................................................... 23 
7.11 Subject Demographics and Baseline Characteristics.................................................. 23 
7.12 Medical and Surgical History..................................................................................... 23 
7.13 Prior and Concomitant Medications........................................................................... 23 
7.14 Measurements of Treatment Compliance................................................................... 23 
7.15 Pharmacokinetic Analysis .......................................................................................... 24 
7.16 Biomarker Analysis .................................................................................................... 25 
7.17 Safety Analyses .......................................................................................................... 25 
7.17.1 
Adverse Events ....................................................................................................... 25 
7.17.2 
Clinical Laboratory Assessments............................................................................ 26 
7.17.3 
Vital Signs............................................................................................................... 26 
7.17.4 
Electrocardiograms ................................................................................................. 27 
7.17.5 
SARA Assessment.................................................................................................. 27 
7.18 Planned Interim Analysis............................................................................................ 27 
8 
GENERAL CONVENTIONS FOR TABLES, LISTINGS, AND FIGURES ...................... 28 
9 
LIST OF TABLES, FIGURES, AND LISTINGS ................................................................ 30 
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1.
INTRODUCTION
This statistical analysis plan (SAP) is consistent with the statistical methods section of the final 
study protocol (Version 3.0 dated 06 June 2022) and includes additional detail of the safety, 
tolerability, and pharmacokinetics (PK) of RGLS8429 summaries to be included in the clinical 
study report (CSR).
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2.
STUDY OBJECTIVES AND ENDPOINTS
2.1 Objectives
The following are the study objectives.
2.1.1
Primary Objective
x
To assess the safety and tolerability of single ascending subcutaneous (SC) doses of 
RGLS8429
2.1.2
Secondary Objectives
x
To identify dose-limiting toxicity (DLT) and to determine the maximum tolerated 
dose (MTD) of a single SC dose of RGLS8429
x
To characterize the PK properties of RGLS8429
2.1.3
Exploratory Objective
x
To characterize multiple potential renal biomarkers
2.2 Endpoints
The following are the study endpoints.
2.2.1
Primary Endpoint
x
Incidence of adverse events (AEs) and change in safety laboratory test results, vital 
signs, scale for the assessment and rating of ataxia (SARA), and electrocardiogram
(ECG) parameters and Holter monitoring over time
2.2.2
Secondary Endpoints
x
Incidence of DLT and determination of MTD
x
PK parameters of RGLS8429 including:

Maximum observed concentration (Cmax)

Time to maximum observed concentration (Tmax)

Area under the concentration–time curve up to 24 hr post dose (AUC0-24)

Area under the concentration–time curve up to the last quantifiable 
concentration (AUC0-t)

Area under the concentration–time curve extrapolated to infinity (AUCinf)

Half-life (t½)

Apparent clearance (CL/F)
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
Apparent Volume of distribution (Vz/F)

Fraction of unchanged RGLS8429 excreted in the urine (fe)

Total amount of unchanged RGLS8429 excreted in the urine (Ae)
2.2.3
Exploratory Endpoints
x
Characterize multiple potential renal biomarkers in urine (e.g., polycystin 1 [PC1], 
polycystin 2 [PC2], neutrophil gelatinase-associated lipocalin [NGAL], kidney 
injury molecule-1 [KIM-1], monocyte chemoattractant protein-1 [MCP-1], beta-2
microglobulin [B2M], and clusterin [CLU]) and in serum (e.g., Cystatin-C, Insulin-
like Growth Factor Binding Protein Acid Labile Subunit [IGFALS], albumin [Alb], 
copeptin [CT-proAVP], N-acetyl-1-methylhistidine, and others).
x
Renal Clearance (CLR)
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3
STUDY DESIGN
3.1 General
This is a Phase 1, randomized, double-blind, placebo-controlled, single ascending dose (SAD)
study in healthy volunteers to evaluate the safety, tolerability, and PK of RGLS8429. A single 
ascending dose of RGLS8429 or placebo will be administered via SC injection to healthy 
volunteers.
Eight subjects will be randomized 3:1 to receive RGLS8429 or placebo in each cohort.
Cohort 1: 0.5 mg/kg RGLS8429 or placebo
Cohort 2: 1.0 mg/kg RGLS8429 or placebo
Cohort 3: 2.0 mg/kg RGLS8429 or placebo
Cohort 4: 4.0 mg/kg RGLS8429 or placebo
In each cohort, 6 subjects will be randomized to receive RGLS8429 and 2 will be randomized to 
receive placebo. Enrollment into subsequent cohorts will be initiated after review of safety (AEs 
and safety laboratory tests) from the prior cohort.
For Cohort 1, the first two subjects will receive RGLS8429 or placebo (randomized such that one
subject receives RGLS8429 and one receives placebo) and this sentinel group will be monitored 
for 48 hrs prior to dosing the remaining subjects in the cohort. 
Subjects will be admitted to the study center on Day -1, one day prior to the day of study drug
administration (Day 1), and will be discharged on Day 2, no less than 6 hrs after the collection of
the 24-hr PK sample. Subjects will be followed for a total of 4 weeks after dosing and will return 
to the site for follow-up visits weekly from Weeks 1 through 4. The sentinel subjects will remain 
in the clinic for 48 hrs after dosing.
Study procedures and assessments with their timing is summarized in Table 3-1.
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Table 3-1: Schedule of Assessments and Procedures 
Screen 1
Treatment
Follow-Up
Week
1
2
3
4/EOS
Days
-28 to -2
-1
1
2
8
15
22
29
Hour
0 2
0.5
1
2
3
4
5
6
8
10
12
16
24
Informed Consent
X
Demographics
X
Medical History
X
Physical Examination 3
X
X
X
X
X
X
X
Weight and Height 4
X
X
X
X
Vital Signs 5
X
X
X
X
X
X
X
X
X
X
X
X
ECGs 6
X
X
X
X
X
X
Hematology 7
X
X
X
X
X
X
X
Chemistry 8
X
X
X
X
X
X
X
Coagulation 9
X
X
X
X
Urinalysis 10
X
X
X
X
X
X
X
Lipids 11
X
X
X
X
X
Pregnancy Test 12
X
X
X
Urine Drug and Alcohol Screen
X
X
X
HBsAg, HCV, HIV Screen
X
Inclusion/Exclusion Review
X
X
Randomization
X
SARA Assessment 13
X
X
X
X
Adverse Events
X
X
X
X
X
Concomitant Medications
X
X
X
X
X
X
X
Study Drug Administration
X
PK Plasma Collection 14, 19
X
X
X
X
X
X
X
X
24-Hr Urine Collection 15, 19
X
Serum biomarkers 16, 19
X
X
Urine biomarkers 17, 19
X
X
Holter monitoring 18
X
Admission to Clinic
X
Discharge from Clinic
X
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Clinic Visit
X
X
X
X
Abbreviations: ECG, electrocardiogram; EOS, end of study or early termination visit; HBsAg, hepatitis B surface antigen; HCV, hepatitis C virus; HIV, human
immunodeficiency virus; PK, pharmacokinetics; SARA=Scale for the Assessment and Rating of Ataxia. 
1. Screening Period within 28 days prior to study drug administration (Baseline = Day -1 or Day 1 when test/procedure performed).
2. Ensure that all Day 1 procedures scheduled to be completed prior to dosing are completed prior to dosing.
3. Complete physical examination at Screening and Week 4/EOS Visit. Limited physical examination at other visits. The limited physical examination will be focused on 
general appearance, the respiratory, neurological, and cardiovascular systems, and subject-reported symptoms 
4. Measure weight at all visits indicated. Height at Screening only. Calculate BMI at Screening Visit.
5. Systolic (SBP) and diastolic (DBP) blood pressure measurements, heart rate (HR), body temperature, and respiratory rate (RR).
6. Obtain a 12-lead ECG during Screening and print and review ECG tracings from the Holter monitor pre-dose and 4 hr, 6 hr, 12 hr, and 24 hr post-dose. Record new or
worsening clinically significant abnormalities as an AE.
7. Full hematology panel including hemoglobin (Hb), hematocrit (Hct), red blood cell (RBC) count, red cell indices, white blood cell (WBC) count including absolute 
cell counts, with differential, and platelet count.
8. Fasting chemistry panel including total protein, glucose, uric acid, creatine kinase (CK), lactate dehydrogenase (LDH), sodium, potassium, chloride, bicarbonate, 
calcium, phosphate, BUN, creatinine (Scr), and hepatic function panel (albumin, total bilirubin, direct bilirubin, ALT, AST, ALP, and gammaglutamyl transferase 
[GGT]).
9. Prothrombin time (PT), activated partial thromboplastin time (aPTT), international normalized ratio (INR), and fibrinogen will be collected on Day -1; Day 1, 4 hr 
post-dose; and Day 2, 24 hr post-dose; and at Week 4/EOS. 
10. Urinalysis panel including specific gravity, pH, protein, glucose, ketones, blood (Hb), leukocyte esterase nitrite, bilirubin, and urobilinogen. If protein, blood, or 
leukocyte esterase nitrate parameters are abnormal a microscopic examination will be completed. 
11. Fasting lipid panel including total cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL).
12. Only female subjects of childbearing potential. A serum pregnancy test will be performed at Screening and urine pregnancy tests will be performed at the remaining 
specified visits.
13. Perform the SARA assessment pre-dose on Day 1 and 4-6 hr post-dose on Day 1; on Day 2, 1 hr after the 24-hr PK sample collection; and at Week 4/EOS Visit.
14. Pre-dose plasma PK sample on Day 1 should be obtained within 60 minutes prior to dosing. Post-dose plasma samples should be obtained within the following time 
margins: collections at 2, 4, 6, and 8 hr ±10 minutes, 12 hr ±30 minutes, and 24 hr ±60 minutes.  
15. 24-hr urine collection for PK analysis will begin immediately after study drug dosing on Day 1 (0-24 hr). The subject should void immediately before dosing (before 
the 24-hr urine collection is started). Last void will be at 24 hours post dose. 
16. Serum sample for exploratory biomarkers (e.g., Cystatin-C, IGFALS, Alb, CT-pro-AVP, N-acetyl-1-methylhistidine, and others). 
17. Collect at least 150 mL (more if possible) of mid-stream free-flow urine for analysis of PC1, PC2, NGAL, KIM-1, MCP-1, B2M, and CLU during Screening and on 
Day -1. These samples must NOT be the first urination of the day (to avoid too much debris in the sample).
18. Start Holter monitoring 1 hr before study drug administration on Day 1 and continue for 30 minutes after the 24-hr PK sample is obtained on Day 2.
19. Unused backup plasma, serum, and urine PK/biomarker samples may also be used for additional exploratory biomarkers analyses.
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3.2 Study Population
32 healthy male or female volunteers 18 to 55 years old and with a body mass index (BMI) 
between 18 and 35 kg/m2 will be randomized.
3.3 Randomization and Treatment Assignments
This study will be double-blind and placebo-controlled. There will be 4 SAD cohorts. Eight
subjects will be enrolled into each of the 4 dosing cohorts. Subjects in each cohort will be 
randomized at a ratio of 3:1 to receive RGLS8429 or placebo. In each cohort, 6 subjects will be 
randomized to receive RGLS8429 and 2 will be randomized to receive placebo. 
ICON will provide a randomization schedule and eligible subjects will be randomized 
sequentially to receive the study drug assigned to the corresponding randomization number. 
Subjects may be randomized once all Screening and Day -1 procedures have been completed and 
eligibility has been confirmed by the Investigator. Details regarding randomization procedures 
will be provided in the study reference manual.
Enrollment into subsequent cohorts will be initiated after review of safety (AEs and safety 
laboratory tests) from the prior cohort.  The investigator or a designee will assign sequential 
subject identification numbers to the subjects as they are screened and enrolled in the study.
An unblinded pharmacist will be responsible for preparing RGLS8429 or placebo and providing 
the study drug to blinded study personnel for administration according to the Pharmacy 
instructions within the Study Reference Manual.
3.4 Determination of Sample Size
Formal sample size calculations were not performed for this study. The dose cohort size is 8 
subjects randomized 3:1 to RGLS8429 versus placebo. The size of each dose cohort was chosen 
to provide sufficient information to allow assessment of the safety and tolerability of RGLS8429.
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4
CHANGES IN THE CONDUCT OF THE STUDY OR PLANNED ANALYSIS
All analyses specified in this SAP are consistent with the final study protocol (Version 3.0, dated 
06 June 2022). Any changes in the analyses provided or any additional analyses performed will 
be documented in the clinical study report (CSR).
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5
QUALITY CONTROL AND QUALITY ASSURANCE METHODS FOR DATA 
ANALYSIS
Case report forms will be reviewed and processed according to the ICON Study Specific 
Procedure SSP DM-38770009.01 Data Management Plan (DMP). The DMP describes eCRF 
data processing, edit checks, data query management, medical dictionary coding, SAE 
reconciliation, data transfers, and data quality review through database lock or any necessary 
reopening of the database. After database lock, the data will be retrieved from the database using 
SAS version 9.4 or higher. Independent programming will be performed for the analysis. 
Statistical review should be conducted on an ongoing basis as each output passes validation after 
lead programmer review or at one-time point only when all outputs have been validated. When 
statistical review is performed and completed, the Lead Statistician, Programming lead and 
Project lead will sign a Biostatistics and Programming Validation and Release Form (refer to 
PRG001-SOP) to verify that all applicable procedures and review steps have been performed and 
authorizing the release of TLFs.
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6
PHARMACOKINETIC AND PHARMACODYNAMIC ASSESSMENTS
6.1 Pharmacokinetic Assessment
Plasma concentrations of unchanged RGLS8429 at different time-points and urine concentrations
collected over the specified interval will be used to derive the following PK parameters using non-
compartmental methods (Phoenix™WinNonlin®, Version 8.0 or later higher, Certara, Princeton, 
NJ)
The following PK parameters will be reported:
Variable
Definition
AUC0-24
Area under the plasma concentration-time curve from time =0 to 24
estimated using the linear/log trapezoidal rule
AUC0-t
Area under the plasma concentration-time curve from time =0 to the last 
quantifiable concentration estimated using the linear/log trapezoidal rule.
AUCinf
Area under the plasma concentration-time curve extrapolated to infinite time 
calculated as:
Where Clast is the last observed quantifiable concentration, and Oz is the 
terminal rate constant for elimination.
Cmax
Maximum observed plasma concentration.
Tmax
Time to Cmax. If the maximum observed concentration value occurs at more 
than 1 time point, Tmax is defined as the first time point with this value.
t½
Terminal elimination half-life in plasma computed as:
Where ln(2) is the natural logarithm of 2.
CL/F
Apparent total clearance of the drug from plasma calculated as:
Vz/F
Apparent volume of distribution during the elimination phase calculated as:
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Variable
Definition
Ae0-24
Total amount of unchanged RGLS8429 excreted in the urine after the first 
dose of RGLS8429 (Day 1):
Where Cut1-t2 is the concentration in urine collected over the interval from 
time 0 to 24 hours and Vut1-t2 is the volume of urine collected over the 
interval from time 0 to 24 hours.
fe0-24
Fraction of the unchanged drug excreted in urine over the interval from time 
t1 to time t2 computed as:
0 24
0 24
Ae
fe
Dose


 
CLR
Renal Clearance computed as:
ܥܮோൌܣ݁଴ିଶସ
ܣܷܥ଴ିଶସ
6.1.1
Treatment of Outliers
Individual concentration-time points, if considered anomalous, may be excluded from the 
analysis at the discretion of the pharmacokineticist following a review of the available 
documentation. Any such exclusion will be discussed with the sponsor and clearly outlined in the 
study report.
Entire individual treatment profiles for a subject may be excluded following review of the 
available documentation and discussion with the sponsor. However, results of analysis with and 
without the excluded profiles may be presented in the study report. Any such exclusion will be 
clearly listed in the study report along with justification for exclusion.
6.1.2
Non-Quantifiable Concentrations
All concentration values reported as no results (not collected or not determined) values will be 
treated as missing. For the calculation of concentration summaries, all concentrations below the 
quantifiable limit (BLQ) will be treated as 0. For the purpose of calculating PK parameters and 
plotting mean and individual concentration-time profiles, BLQ values will be treated as 0 prior to 
the first measurable concentration. After the first measurable concentration, subsequent BLQ 
values (or mean concentration values that are BLQ) are treated as missing.
6.2 Biomarker Assessment
Collect mid-stream free-flow urine for analysis of urine exploratory biomarkers (e.g., PC1, PC2, 
NGAL, KIM-1, MCP-1, B2M and CLU) during Screening and Day -1 and collect serum samples 
during Screening and Day -1 for serum exploratory biomarkers (e.g., Cystatin-C, IGFALS, Alb, 
CT-pro-AVP, N-acetyl-1-methylhistidine, and others). 
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7
STATISTICAL METHODS
7.1 General
The statistical analysis will be conducted following the principles specified in the International 
Council for Harmonisation (ICH) Topic E9 Statistical Principles for Clinical Trials 
(CPMP/ICH/363/96).
All statistical analyses will be performed using the statistical software SAS Grid/ SAS Linux -
SAS®, Version 9.4 or newer and any exceptions will be detailed in the CSR.
All results collected in the database will be presented in listings. All continuous data will be 
listed with the same precision as presented in the database. 
Unless otherwise noted, continuous variables will be summarized descriptively using the number 
of non-missing observations (n), arithmetic mean (mean), standard deviation (SD), coefficient of 
variation (%CV), first (Q1) and third quartiles (Q3), median, minimum, and maximum; 
categorical variables will be summarized using the frequency count and the percentage of 
subjects in each category.
In the data listings, study day relative to dose of study drug will be presented. Study day relative
to dose will be calculated as event date – dose date (+1 if event date ≥ dose date).
Baseline will be the latest available measurement (scheduled or unscheduled assessment) prior to 
the study drug administration in the study.
Subjects will be analyzed based on the actual treatment received.
For numerical variables, change from baseline will be calculated as (Post baseline value –
Baseline value).
For safety summaries, the unscheduled and repeat assessments will not be included in descriptive 
statistics; however, all results will be included in the data listings.
All data will be summarized by cohort and treatment group, pooled RGLS8429, and pooled 
placebo unless otherwise specified.
7.2 Handling of Dropouts or Missing Data
There will be no imputations for missing data other than that implicit by ignoring missing data 
when using the trapezoidal rule for the calculation of AUCs. For all variables, only the observed 
data from the subjects will be used in the by-visit summaries.
The following imputation dates for AE and concomitant medication will only be applied for 
determination of TEAEs and prior/concomitant medication.
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7.2.1
Handling of missing/ incomplete dates for Adverse Event
Imputation rules for missing or partial AE start date are defined below:
a) If only Day of AE start date is missing:
If the start date has month and year but day is missing, the first day of the month will be imputed.
x
If this date is earlier than the first dose date, then the first dose date will be used instead.
x
If this date is later than the stop date (possibly imputed), then the stop date will be used 
instead.
b) If Day and Month of AE start date are missing:
If the start date has year, but day and month are missing, the 1st of January will be imputed.
x
If this date is earlier than the first dose date, then the first dose date will be used instead.
x
If this date is later than the stop date (possibly imputed), then the stop date will be used 
instead.
c) If Year of AE start date is missing:
If the year of AE start is missing or AE start date is completely missing then imputation will not 
be done and this AE will be considered as TEAE.
7.2.2
Handling missing or partial Prior/Concomitant Medication Dates
Missing or partial medication start date:
a. If only Day is missing, the first day of the month will be assumed.
b. If Day and Month are both missing, the first day of the year will be assumed.
c. If Day, Month and Year are all missing, the day before the first dose date will be assumed.
Missing or partial medication stop date:
a. If only Day is missing, the last day of the month will be assumed.
b. If Day and Month are both missing, the last day of the year will be assumed.
c. If Day, Month and year are all missing, ‘continuing’ status to stop date will be assigned.
7.3 Multiple Comparisons and Multiplicity
There will be no multiple comparison adjustment for various analyses. 
No formal statistical tests will be conducted to assess the safety or tolerability of RGLS8429.
7.4 Adjustments for covariates
Not applicable.
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7.5 Multicenter Studies
This is a single-center study.
7.6 Examination of Subgroups
No subgroup analyses are planned.
7.7 Coding dictionaries
Medical history, AEs and medical procedures will be coded using the Medical Dictionary for 
Regulatory Activities (MedDRA, Version 24.0 or higher). Medications will be coded using the 
World Health Organization dictionary of medical codes (WHO Global B3 format - March 2021).
7.8 Analysis Populations
7.8.1
Screened Population
Screened population includes all subjects who signed the informed consent form 
7.8.2
Randomized Population
Randomized population includes all subjects who are randomized into the study.  
7.8.3
Safety Population
The Safety Population consists of all subjects who received a dose of study drug. Subjects will 
be analyzed based on treatment received.
7.8.4
Pharmacokinetic Full Population
The PK Full Population consists of all subjects who received a dose of RGLS8429 and have at 
least 1 concentration data point. Subjects will be analyzed based on dose received.
7.8.5
Pharmacokinetic Evaluable Population
The PK Evaluable Population consists of all subjects who received a dose of RGLS8429 and 
have at least one PK parameter that can be reliably estimated. Subjects will be analyzed based on 
dose received.
7.8.6
Biomarkers (Pharmacodynamic (PD)) Population
The Biomarker (PD) Population consists of all screened subjects who have at least one screening 
and/or pre-dose value for the biomarker parameters.
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7.9 Subject Accountability
Summaries of analysis populations and subject disposition will be presented by cohort and 
overall. It will contain the following information:
x
Number of subjects screened
x
Number of subjects randomized
x
Number and percent of subjects who completed the study
x
Number and percent of subjects who discontinued early and reason for early 
discontinuation
x
Number and percent of subjects in the Safety, PK Full, PK Evaluable, and PD
Populations
Percentage will be calculated based on number of subjects in Randomized Population except for 
number of subjects screened.
Subject disposition data, exclusions from the analysis populations, eligibility criteria satisfaction 
and consent information will be presented in listings for the Randomized Population.
7.10 Protocol Deviations
All protocol deviations will be identified prior to database lock. Major protocol deviations will 
be listed. In addition, a separate listing of all inclusion and exclusion deviations will be provided.
7.11 Subject Demographics and Baseline Characteristics
Subject demographics (age, sex, race, and ethnicity) and baseline characteristics (height, weight,
BMI, and eGFR) will be summarized descriptively based on the Randomized Population. Subject
demographics and baseline characteristics will be listed.
7.12 Medical and Surgical History
The medical and surgical history data will be coded using the MedDRA, Version 24.0 or higher.
and will be listed and summarized using the Randomized Population.
7.13 Prior and Concomitant Medications
All prior and concomitant medications will be coded using the WHO Global B3 format - March 
2021. Concomitant Medications will be listed and summarized using the Safety Population. Prior 
Medications will be listed only for Randomized Population. A separate listing will be provided 
for concomitant procedures for Safety population.
7.14 Measurements of Treatment Compliance
A listing of treatment compliance will be provided for eCRF collected exposure data e.g expected
dose, actual dose, number of injections etc. for the Randomized Population.
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7.15 Pharmacokinetic Analysis
The PK Full Population will be used for all concentration (plasma and urine) listings, summary 
tables, and graphs. The PK Evaluable Population will be used for PK parameters listing (plasma 
and urine), PK parameters summary (plasma and urine), and statistical analysis.
The individual sampling and subject concentration-time for plasma drug concentrations of 
RGLS8429 will be listed and displayed graphically on linear and semi-log scales. The drug 
concentrations will be summarized descriptively by RGLS8429 cohort and nominal time point in 
tabular and graphical formats (linear and semi-log scales). Linear and semi-logarithmic plots of 
the individual plasma concentration by actual sampling time for each RGLS8429 cohort will be 
provided by subject (one subject per page). Overlay of individual plasma concentration profiles 
over time will be provided by RGLS8429 cohort. Plots of mean (SD) plasma concentration 
profiles versus time will be presented with all cohorts superimposed on the same plot.
All PK parameters will be listed. Summary statistics will be calculated for the plasma
RGLS8429 PK parameters for each dose cohort using descriptive statistics to include, as 
appropriate, n, mean, SD, CV, Q1 and Q3, minimum, median, maximum, geometric mean and 
geometric %CV. For Tmax, summary statistics will be described by n, minimum, median, and 
maximum only.
The total amount of RGLS8429 excreted in urine will also be assessed, where feasible. Urine 
pharmacokinetic parameters (CLR., fe, Ae) will be listed for each subject by RGLS8429 cohort
and summarized using descriptive statistics. Individual urine concentrations and volumes will be 
listed. In addition, urine samples will be qualitatively screened for potential RGLS8429 
catabolites.
Additional PK parameters may be estimated as appropriate. The parameters stated above will be 
reported if they are estimable and will be discussed in the CSR. When necessary, interpolation or 
λz extrapolation may be used to estimate partial AUCs. The value of AUC%extrap will be less than 
or equal to 20% for the AUCinf to be considered well estimated. If this proportion is > 20%, then 
the values of AUCinf will be treated with caution. When AUCinf is calculated and tabulated, the 
AUC%extrap will also be tabulated. Where AUC%extrap >20%, AUCinf and all related parameters 
(i.e. CL/F and Vz/F) will be presented but excluded from the calculation of summary statistics. 
All values excluded from the summaries should be flagged in the individual listings with an 
explanation for the exclusion.
The apparent terminal phase rate-constant (Oz) will be estimated by log-linear regression of the 
concentration-time data associated with this phase. The decision as to which data points describe 
the terminal phase will be reached by inspecting the semi-logarithmic plot of the data, only 
considering concentrations at time points beyond Tmax. A minimum of three data points will be 
used for the estimation of Oz preferably covering a time span of at least 2 half-lives. Poorly 
estimated Oz (i.e. if a time span of at least 2 half-lives is not used or r2adj d 0.80), the 
corresponding t½ and other parameters derived using Oz (i.e. AUCinf, CL/F, Vz/F, and t½) will be 
flagged in the report. Summary statistics of these parameters (i.e. AUCinf, CL/F, Vz/F, and t½)
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will be reported with and without these outliers (i.e., with and without parameters where Oz was 
poorly estimated).
Dose Proportionality Assessment:
Dose proportionality will be explored using the power model for Cmax, AUCinf, AUC0-24 and 
AUC0-t as data permit for the PK evaluable population.
A power model will be fit separately to describe the relationship between pharmacokinetic 
parameter (Cmax, AUCinf, AUC0-24 and AUC0-t) and actual dose in mg using the least-squares 
linear regression model as follows:
ln(PK parameter) = α + β ln(dose) + ε.
This linear model will be used to provide a 95% confidence interval (CI) for β (slope).
From each pharmacokinetic parameter model, the intercept of the regression line α and the slope 
of the regression line β will be presented along with the 95% CI of the slope. Actual dose of drug 
and not dose level will be used in analysis. A minimum of 3 non-missing PK parameter values 
per dose cohort must be available to estimate dose proportionality with the power model.
7.16 Biomarker Analysis
The Biomarker (PD) Population will be used for all biomarker analyses. Baseline (the average of 
screening and Day -1 samples) biomarker data will be listed and summarized using descriptive 
statistics. Three summaries will be presented: Summary of Screening (Day -28 to Day-2),
Summary of Day -1 and Summary of the average of Screening and Day-1 samples (Baseline).
7.17 Safety Analyses
All Safety analysis will be presented using the Safety Population. No formal statistical tests will 
be conducted to assess the safety or tolerability of RGLS8429.
Tables will be presented by cohort and treatment group, pooled RGLS8429 and pooled placebo.
7.17.1 Adverse Events
All AE summaries will include TEAEs. TEAEs or TESAEs are those which occur or worsen 
after the administration of study drug until the Day 29 or EOS Visit whichever occurs later.
All AEs will be classified by primary system organ class (SOC) and preferred term (PT) 
according to the MedDRA Version 24.0 or higher. Data collected in CRF along with duration of 
AE will be presented in listing. Duration of AE will be calculated as stop date – start date.
The overall incidence of TEAEs (number and percentage of subjects) as well as the number of 
events will be summarized. It includes severity, relationship to study drug of TEAE and SAEs, 
TEAEs leading to study discontinuation, life-threatening SAEs, and SAEs resulting in death.
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The TEAEs will be summarized and tabulated at both the subject (n [%] of subjects) and event 
(number of events) level:
x
TEAEs by SOC and PT 
x
TEAEs by SOC, PT, and maximum reported severity
x
TEAEs by SOC, PT, and relationship to study drug
x
TESAEs by SOC and PT
For the incidence at the subject level by SOC and PT, if a subject experiences more than 1 event 
within the same SOC and PT, only 1 occurrence will be included in the incidence.
For the incidence at the subject level by SOC, PT, and severity, if a subject experiences more 
than 1 event within the same SOC and PT, only the most severe occurrence will be included in 
the incidence.
For the incidence at the subject level by SOC, PT, and relationship to study drug; if a subject
experiences more than 1 event within the same SOC and PT, only the most closely related 
occurrence will be included in the incidence.
The incidence of DLTs will be summarized by cohort and treatment group.
Any SAEs, AEs with outcome of death and AEs resulting in discontinuation of study will be 
listed separately.
7.17.2 Clinical Laboratory Assessments
Clinical laboratory data will be summarized using System International (SI) units and the Safety 
Population will be used.
Observed and change from baseline of continuous clinical laboratory values (serum chemistry,
coagulation panel, hematology, lipids, and urinalysis) will be summarized for each parameter at
each visit/timepoints using descriptive statistics.
Shift from baseline to worst post-baseline laboratory findings based on normal range criteria will 
also be summarized.
A listing of all clinical laboratory data for each subject at each visit will be presented. Laboratory 
values outside the reference range will be flagged with ‘L’ for low and ‘H’ for high in the data 
listings and a list of abnormal values will be presented.
For female subjects, serum/urine pregnancy test results during the study will be presented in the 
individual subject data listings.
7.17.3 Vital Signs
Observed and change from baseline vital signs values will be summarized at each timepoint. 
Vital signs data will be listed by subject at each timepoint.
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7.17.4 Electrocardiograms
Shift tables will be presented that summarize the overall ECG interpretation as normal, abnormal 
not clinically significant, or abnormal clinically significant from baseline to each post-baseline 
time point.
Individual ECG data will be listed.
A separate statistical analysis plan will be prepared for the Holter monitoring data following an
Early Precision QT (EPQT) analysis. This is not included within this statistical analysis plan. 
7.17.5 SARA Assessment
Scale for the Assessment and Rating of Ataxia (SARA) scores for individual item will be 
summarized descriptively. Observed and change from baseline in SARA individual and total 
scores will be summarized at each timepoint.
Total SARA score for each subject will be calculated including score for gait, stance, sitting, 
speech disturbance and mean score of finger chase, nose-finger test, fast alternating hand 
movements and heel-shin slide.
7.18 Planned Interim Analysis
There is no formal interim analysis planned.
However, ongoing assessments of the PK, safety and tolerability, will be performed for dose 
escalation meetings.
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8
GENERAL CONVENTIONS FOR TABLES, LISTINGS, AND FIGURES
Tables and listings will be presented in landscape mode with minimum of 3/4” bound edge 
margin and 3/8” other margins on 8.5” x 11” paper.
Times new roman font size of no less than 8 point will be used for tables and listings.
A source line will be included on the bottom of each page of all tables and listings. It will contain 
the SAS code program name and the run date and time.
Each variable is recorded to a specific number of decimal places. If the raw data is presented 
with varying precision, then the least precise value will be considered as the data precision.
For summary tables, unless otherwise specified, the number of decimal places provided in the 
tables and listings will be based on the accuracy of the least accurate value in the raw data as 
follows:
n
Integer
Arithmetic mean
1 decimal place more than the least accurate number in the raw data
SD
2 decimal places more than the least accurate number in the raw data
%CV
2 decimal places
Geometric mean
1 decimal place more than the least accurate number in the raw data
Geometric CV
2 decimal places
Median
1 decimal place more than the least accurate number in the raw data
Quartiles (Q1 and Q3)
1 decimal place more than the least accurate number in the raw data
Minimum
same number of decimal places as raw data
Maximum
same number of decimal places as raw data
Confidence interval
same number of decimals as the associated statistic
Percentage
1 decimal place
Geometric mean ratio
2 decimal places
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Statistical Analysis Plan Final v1.0
19 Sep 2022
CONFIDENTIAL
Page 29 of 32
The sample column layouts are given below:
RGLS8429 
0.5 mg/kg 
(N=xx) 
RGLS8429 
1.0 mg/kg 
(N=xx) 
RGLS8429 
2.0 mg/kg 
(N=xx) 
RGLS8429 
4.0 mg/kg 
(N=xx) 
 
Pooled 
RGLS8429 
(N=xx) 
Pooled 
Placebo  
(N=xx) 
x
For PK tables placebo will not be presented
The listing layouts are given below:
Cohort/ Treatment 
Subject Number/ Age/ Sex/ 
Race 
xxxx 
xxxx 
xxxx 
 
 
 
 
 
Cohort 1/ RGLS8429 0.5 mg/kg  
XXXX-XXXX/ xx/ X/ X 
 
 
 
Cohort 1/ Placebo 
XXXX-XXXX/ xx/ X/ X 
 
 
 
Cohort 2/ RGLS8429 1.00 mg/kg  
XXXX-XXXX/ xx/ X/ X 
 
 
 
Cohort 2/ Placebo 
XXXX-XXXX/ xx/ X/ X 
 
 
 
....... 
 
 
 
 
 
 
 
 
 
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Regulus Therapeutics Inc.
Protocol. No. RGLS8429-01
ICON Project No. 3877/0009 
Statistical Analysis Plan Final v1.0
19 Sep 2022
CONFIDENTIAL
Page 30 of 32
9
LIST OF TABLES, FIGURES, AND LISTINGS
Table/Figure 
Number
Table/Figure Name
Section 14.1
Demographic and Subject Characteristics Data Summaries
Table 14.1.1
Summary of Subject Disposition (Screened Population)
Table 14.1.2
Analysis Populations 
Table 14.1.3
Demographic and Baseline Characteristics (Randomized Population)
Table 14.1.4
Medical and Surgical History Summary (Randomized Population)
Section 14.2
Pharmacokinetic/Biomarker Data Summaries
Section 14.2.1
Pharmacokinetic Data Summaries
Table 14.2.1.1
Summary of RGLS8429 Plasma Concentrations (units) by Timepoint 
(PK Full Population)
Table 14.2.1.2
Summary of RGLS8429 Plasma PK Parameters (PK Evaluable 
Population)
Table 14.2.1.3
Summary of RGLS8429 Plasma PK Parameters after excluding Outliers
(PK Evaluable Population)
Note: Outliers are included (e.g., estimates of T1/2 if a time span of at 
least 2 half-lives is not used or r2adj d 0.80)
Table 14.2.1.4
Summary of RGLS8429 Urine PK Parameters (PK Evaluable 
Population)
Table 14.2.1.5
Statistical Analysis of Dose Proportionality of Pharmacokinetic 
Parameters (PK Evaluable Population)
Figure 14.2.1.1
Mean (+/-SD) of Plasma Concentration of RGLS8429 -Time Profiles by 
Treatment Group (PK Full Population) 
Note: Present both linear and semi-log
Figure 14.2.1.2
Plot of RGLS8429 Dose-Proportionality Regression (PK Evaluable 
Population)
Section 14.2.2
Biomarker Data Summaries
Table 14.2.2.1
Summary Statistics of Urine Biomarker Variables (Biomarker (PD) 
Population)
Section 14.3
Safety Data Summaries
Section 14.3.1
Displays of Adverse Events
Table 14.3.1.1
Overall Summary of Treatment-Emergent Adverse Events (Safety 
Population)
Table 14.3.1.2
Summary of Treatment-Emergent Adverse Events by System Organ 
Class and Preferred Term (Safety Population)
Table 14.3.1.3
Summary of Treatment-Emergent Adverse Events by Maximum Severity 
Grade (Safety Population)
Table 14.3.1.4
Summary of Treatment-Emergent Adverse Events Related to Study Drug 
(Safety Population)
Table 14.3.1.5
Summary of Treatment-Emergent Serious Adverse Events (Safety 
Population)
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Regulus Therapeutics Inc.
Protocol. No. RGLS8429-01
ICON Project No. 3877/0009 
Statistical Analysis Plan Final v1.0
19 Sep 2022
CONFIDENTIAL
Page 31 of 32
Table 14.3.1.6
Summary of Incidence of Dose-Limiting Toxicity (Safety Population)
Table 14.3.1.7
Concomitant Medication Summary (Safety Population)
Section 14.3.2
Listings of Deaths, Other Serious and Certain Significant Adverse 
Events
Table 14.3.2.1
Listing of Deaths (Safety Population)
Table 14.3.2.2
Listing of Serious Adverse Events (Safety Population)
Table 14.3.2.3
Listing of Adverse Events Resulting in Discontinuation of Study (Safety 
Population)
Section 14.3.4
Abnormal Laboratory Value Listing
Table 14.3.4
Listing of Abnormal Laboratory Results (Safety Population)
Section 14.3.5
Additional Safety Data Summaries
Table 14.3.5.1
Summary Statistics of Absolute and Change from Baseline for Clinical 
Laboratory Parameters (Safety Population)
Note: continuous tests only
Table 14.3.5.2
Shift Table for Clinical Laboratory Parameters (Safety Population)
Table 14.3.5.3
Summary Statistics of Absolute and Change from Baseline for Vital 
Signs (Safety Population)
Table 14.3.5.4
Shift Table for Electrocardiogram Interpretations (Safety Population)
Table 14.3.5.5
Summary Statistics of Absolute and Change from Baseline in Scale for 
the Assessment and Rating of Ataxia (SARA) (Safety Population)
Listing Number
Listing Name
Section 16.2.1
Discontinued Subjects
Listing 16.2.1.1
Screening Outcome (Screened Population)
Listing 16.2.1.2
Subject Disposition (Randomized Population)
Note: Randomization and study completion 
Section 16.2.2
Protocol Deviations
Listing 16.2.2
Protocol Deviations (Safety Population)
Section 16.2.3
Subjects Excluded from Analysis
Listing 16.2.3
Analysis Populations
Section 16.2.4
Demographic Data
Listing 16.2.4.1
Demographics and Baseline Characteristics (Randomized Population)
Note: including Screening Height, Weight, eGFR
Listing 16.2.4.2
Consent Information (Randomized Population)
Listing 16.2.4.3
Medical and Surgical History (Randomized Population)
Listing 16.2.4.4
Prior Medications (Randomized Population)
Listing 16.2.4.5
Concomitant Medications (Safety Population)
Listing 16.2.4.6
Prior and Concomitant Procedures (Safety Population)
Section 16.2.5
Compliance and/or Drug Concentration Data
Listing 16.2.5.1
Study Drug Dosing Record (Randomized Population)
Listing 16.2.5.2
Individual RGLS8429 Plasma Concentrations (PK Full Population)
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Regulus Therapeutics Inc.
Protocol. No. RGLS8429-01
ICON Project No. 3877/0009 
Statistical Analysis Plan Final v1.0
19 Sep 2022
CONFIDENTIAL
Page 32 of 32
Listing 16.2.5.3
Individual RGLS8429 Urine Concentrations (PK Full Population)
Figure 16.2.5.1
Individual RGLS8429 Plasma Concentration-Time (PK Full Population) 
Note: For each subject, present the individual concentration in linear and 
semi-log scale.
Figure 16.2.5.2
Spaghetti Plot of RGLS8429 Overlay of Individual RGLS8429 Plasma 
Concentrations-Time by Dose (PK Full Population) 
(Note: the individual concentration will be presented in the same plot for 
linear and semi-log scales.)
Section 16.2.6
Individual Pharmacokinetic/Biomarkers Data
Section 16.2.6.1
Individual Pharmacokinetic Data
Listing 16.2.6.1.1
RGLS8429 Individual Plasma Pharmacokinetic Parameters (PK Evaluable 
Population)
Listing 16.2.6.1.2
RGLS8429 Individual Urine Pharmacokinetic Parameters (PK Evaluable 
Population)
Section 16.2.6.2
Individual Biomarkers
Listing 16.2.6.2
Individual Biomarker Data (Biomarker (PD) Population)
Section 16.2.7
Adverse Event Listings
Listing 16.2.7
All Adverse Events (Safety Population)
Section 16.2.8
Individual Laboratory Measurements by Subject
Listing 16.2.8.1
Normal Ranges for Laboratory Data (Safety Population)
Listing 16.2.8.2
Clinical Laboratory Data by Category (Safety Population)
Listing 16.2.8.3
Vital Signs (Safety Population)
Listing 16.2.8.4
12-lead Electrocardiogram Results (Safety Population)
Listing 16.2.8.5
Serology Test Results (Safety Population)
Listing 16.2.8.6
Alcohol Screen Test Result (Safety Population)
Listing 16.2.8.7
Urine Drugs Screen Result (Safety Population)
Listing 16.2.8.8
Pregnancy Test Results (Safety Population)
Listing 16.2.8.9
Scale for the Assessment and Rating of Ataxia Results (Safety 
Population)
Listing 16.2.8.10
COVID-19 Impact (Safety Population)
Listing 16.2.8.11
Comments (Safety Population)
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