Vitamin D Levels in Subjects With Vitamin D Deficiency Following Topical Application of Three Doses of Vitamin D Ointment - A Proof of Concept Study
Phase: PHASE1
Conditions: Vitamin D Deficiency
Sponsor: University of Minnesota

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
 
 
 
VITAMIN D LEVELS IN SUBJECTS WITH VITAMIN D DEFICIENCY 
FOLLOWING TOPICAL APPLICATION OF THREE DOSES OF 
VITAMIN D GEL – A PROOF OF CONCEPT STUDY 
 
Protocol No: VITD-001 
Version: 2.0. 
Date: September 16, 1, 2016 
 
 
 
SPONSOR-INVESTIGATOR CONTACT 
 
 
Kevin A. Peterson, MD, MPH 
Title: Professor and Director,  
Center of Excellence in Primary Care 
University of Minnesota Medical 
School. 
Phone: +1-651-357-2522 
peter223@umn.edu 
 
 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
1. 
SYNOPSIS 
Study Title: 
Vitamin D Levels in Subjects with Vitamin D Deficiency 
Following Topical Application of Vitamin D Gel – A Proof of 
Concept Study 
Protocol Number: 
VITD-001 
Development Phase: 
Phase 1  
Indication: 
Subjects with known vitamin D deficiency  
(<20 μg/L Total 25-OH Vitamin D)  
Study Drugs: 
(Including test, 
comparator, dosage 
form, dosing regimen 
and route) 
A novel vitamin D3 (cholecalciferol) is provided as a topical gel using 
ingredients that are identified in the USP/NF and/or designated in the 
list of Inactive Ingredients in FDA Approved Drugs.  The ingredients in 
the topical gel are well known in the industry and have references in the 
pharmaceutical literature and some have available FDA drug master 
file information.  A single applicator contains 0.75g of gel, containing 
150,000 international units of vitamin D3 (cholecalciferol) to be 
applied to the upper arm.  One dose consists of two applicators, one 
applied to each upper arm, for a total of 300,000 international units per 
dose. 
Subjects with known vitamin D deficiency of ≤20 ng/ml total 25-OH 
Vitamin D (50 nmol/liter) will be consented and enrolled. They will 
have a blood draw prior to dosing on Day 1 (to obtain a baseline 
vitamin D, calcium, albumin, and parathyroid hormone level) and will 
receive 300,000 units of topical vitamin D3.  On Day 7, subjects will 
return to the clinic for a blood draw to assess Vitamin D level and to 
check for any signs of skin rash and/or reaction.  If a subject has a skin 
rash and/or reaction prior to Day 7, he/she will be advised to return to 
the clinic for assessment as soon as possible. Any rash and/or skin 
reaction may be photographed and diagnosed. A second dose of 
300,000 units of topical Vitamin D will be applied by the nurse at the 
day 7 visit. Subjects will return on Day 14 for a third dose and a third 
blood draw.  A final blood draw to assess for Vitamin D level will be 
done on day 21.  
A study nurse or physician will apply each dose of topical Vitamin D. 
Prior to application of the gel, 2 strips of paper tape will be applied to 
the subject’s upper arms. One strip will be applied approximately 2 
inches below the subject’s shoulder and the second strip will be applied 
approximately 2 inches above the subject’s elbow.  The entire contents 
of one syringe (0.25 mL of gel) will be applied onto the subjects arm in 
a straight line, from approximately 1 inch below the first strip of paper 
tape to approximately 1 inch above the second strip of paper tape.  The 
subject will be instructed to gently rub the gel into their upper arm with 
their fingertips, while staying between the two pieces of paper tape, for 
approximately 2 minutes or until the gel has been absorbed.  After 
completing the application process on both arms, the subject will wash 
his/her hands thoroughly. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
No. Subjects: 
20 subjects will participate.  If the change from baseline in 25-OH 
vitamin D is consistently less than 10 µg/L in the first 10 subjects then 
the study will be discontinued.  
No. Centers: 
The study will be conducted at the University of Minnesota (single 
center) 
Study Duration: 
The maximum duration of treatment is 14 days (study Days 0, 7 and 
14). Subjects will be followed for change in vitamin D blood level and 
for safety until Study Day 21.  It is anticipated that the study will last 4 
months to enroll and complete dosing and assessment of 20 patients. 
 
Objectives of the 
Study: 
The primary objective of the study is to determine the change in 
vitamin D blood levels following 3 doses of topical vitamin D3 
treatment.  The secondary objective is to assess the safety (based on 
skin rash and/or other reaction, as well as calcium and parathyroid 
hormone levels) following the application of 3 doses of vitamin D3 gel. 
Study Endpoints: 
(Primary and 
Secondary) 
The primary endpoint is to establish the average change in vitamin D 
blood levels following 3 doses of treatment with vitamin D gel. The 
secondary study endpoint is the number of subjects who have any skin 
rash and/or other skin reaction following treatment with the vitamin D 
gel. 
Study Design: 
The study is a pre vs. post open-label trial to assess the change in 
vitamin D blood levels following treatment with 3 doses of topical 
vitamin D gel.  Subjects with known vitamin D deficiency will be 
consented and enrolled. They will have a blood draw prior to dosing on 
Day 0 (to obtain a baseline vitamin D level) and will receive topical 
vitamin D3 on Days 0, 7 and 14. On Days 7, 14, and 21 subjects will 
return to the clinic for a post-dose blood draw and to be assessed for 
any signs of skin rash and/or reaction.  If a subject has a skin rash 
and/or reaction at any time during the study, he/she should return to the 
clinic for assessment as soon as possible. Any rash and/or skin reaction 
will be photographed and diagnosed.  
Eligibility Criteria 
(Inclusion and 
Exclusion): 
Inclusion criteria: 
1. Known vitamin D deficiency based on blood work 
obtained prior to study consent defined as serum 25-
hydroxyvitamin D less than or equal to 20 ng/mL within 
30 days prior to study entry. (Normal ≥ to 30 μg/L total 25-
OH Vitamin D) 
2. Adult, age 18 to 85 years, male or female 
3. If on oral vitamin D therapy, have been on a stable dose 
for the previous 90 days. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
Eligibility Criteria 
(continued): 
Exclusion criteria: 
1. History of chronic liver disease with elevated liver 
function tests, chronic kidney disease (stage 3 or greater, 
eGFR <60 mL/min), uncontrolled thyroid disease 
(elevated thyroid function tests), primary or secondary 
hyperparathyroidism, hypercalcemia, or multiple 
endocrine neoplasia. 
2. Hypercalcemia defined as either elevated corrected serum 
calcium >10.2 mg/dL) measured within 3 months prior to 
study. 
3. eGFR  <60 mL/min within 3 months prior to study. 
4. Active cancers 
5. Women who are pregnant or breastfeeding. 
6. Individuals who are unable to give informed consent 
7. Individuals with psoriasis, active eczema or other skin 
disease, or who are currently receiving treatments or 
medications for skin disease. 
8. Individuals who do not agree to refrain from using 
tanning beds for the duration of the study. 
9. Individuals who do not agree to avoid submerging the gel 
site in water for 8 hours after gel application. 
10. Family members of the study faculty or staff. 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
Study Procedures: 
(Including 
pharmacokinetic (PK) 
sampling times) 
The following procedures will be performed during this study: 
 Chart review to assess for potential eligibility 
 Day 0 
o Subject informed consent 
o Measurement of height and weight and examination of 
skin for rash/appearance to confirm eligibility 
o Record Fitzpatrick skin test score. 
o Blood draw for baseline vitamin D levels (total vitamin 
D 25-OH and vitamin D3)  
o Blood draw for serum creatinine (for GFR calculation), 
albumin (for corrected calcium) and parathyroid 
hormone 
o Application of Vitamin D gel by study staff to inside of 
each arm 
o Instruct subject on assessment of skin. If any rash or 
other skin reaction appears, contact the clinic as soon 
as possible for an assessment. 
 
Day 7 
o Assessment of skin for rash or other reaction 
o Draw blood for Ca, vitamin D level. 
o Application of Vitamin D gel by study staff to inside of 
each arm.  
 
Day 14 
o Assessment of skin for rash or other reaction 
o Draw blood for Ca, vitamin D level. 
o Application of Vitamin D gel by study staff to inside of 
each arm.  
 
Day 21 
o Assessment of skin for rash or other reaction 
o Blood draw for post dose vitamin D, calcium, and 
parathyroid hormone level. 
 
Safety 
All subjects will be monitored for skin rash or other reactions. Any rash 
and/or reaction will be photographed by study staff and 
assessed/diagnosed by a physician. 
Patient will have serum calcium followed each week during the study. 
Although it is unlikely that calcium will change with an increase in 
topical Vitamin D, hypercalcemia presents a potential theoretic risk if 
absorption is high or unexpected variability in vitamin D absorption 
exists.   
Laboratory 
Parameters/Analysis: 
Laboratory analyses will consist of 25-hydroxyvitamin D blood levels 
(D2 and D3) and parathyroid hormone levels.  For safety, an albumen 
adjusted calcium determination will be measured on day 0, 7, 14, and 
21.   
Total Blood Volume: 
Approximately 5 mL will be required at day 0, 7, 14, and 21 to assess 
for vitamin D parameters and safety. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
Sample Size 
Determination: 
(If applicable) 
The study will have 20 subjects who are evaluable for changes in 
vitamin D blood levels.   
Statistical Analyses: 
(Brief Description) 
Two sample paired t test of pre and post total vitamin D and 
parathyroid hormone with an alpha of .05.  We do not expect any 
significant change in adjusted calcium.  

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
2. 
STUDY SCHEDULE 
Table 1: 
Schedule of Events 
 
 
Testing Visits 
Clinical/Study Assessment 
Prescreening 
Day 0 
Day 7 
Day 14 
Day 21 
Chart review for potential 
eligibility 
 
X 
 
 
 
 
Physical Exam of skin* 
∙Skin integrity assessment 
∙Fitzpatrick skin score 
 
 
X** 
 
X** 
 
X** 
 
X** 
Blood Draw: 
∙ Serum 25-OH vitamin D 
∙ Serum calcium 
 
 
 
X 
 
X 
 
X 
 
X 
∙ Serum creatinine (eGFR) 
∙ Serum albumin  
 
 
X 
 
 
 
∙ Serum parathyroid hormone 
 
X 
 
 
X 
Topical Treatment Application 
 
X 
X 
X 
 
* Subjects will be instructed to contact the site as soon as possible if he/she notices a skin rash or any other reaction. 
**A digital photograph may be taken to record any signs of irritation or reaction.  

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
3. 
TABLE OF CONTENTS 
1. 
TITLE PAGE ........................................................... Error! Bookmark not defined. 
2. 
SYNOPSIS ..............................................................................................................2 
3. 
STUDY SCHEDULE .............................................................................................7 
4. 
TABLE OF CONTENTS ......................................................................................8 
5. 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ..................11 
6. 
INTRODUCTION................................................................................................12 
6.1 
Vitamin D Deficiency ...............................................................................12 
6.2 
Study Rationale and Selection of Dose ...................................................14 
6.2.1 
Rationale for Study ...................................................................14 
6.2.2 
Rationale for Selected Dose ......................................................14 
7. 
STUDY OBJECTIVES ........................................................................................16 
8. 
INVESTIGATIONAL PLAN .............................................................................17 
8.1 
Overall Study Design ...............................................................................17 
8.2 
Study Duration .........................................................................................17 
9. 
STUDY POPULATION ......................................................................................18 
9.1 
Inclusion Criteria .....................................................................................18 
9.2 
Exclusion Criteria ....................................................................................18 
9.3 
Criteria for Withdrawal ..........................................................................18 
9.4 
Replacement of Subjects..........................................................................19 
10. 
TREATMENT OF SUBJECTS ..........................................................................20 
10.1 
Treatment Assignment ............................................................................20 
10.2 
Treatment Administration ......................................................................20 
11. 
STUDY DRUG MATERIALS AND MANAGEMENT ...................................21 
11.1 
Description of Study Drug ......................................................................21 
11.2 
Packaging and Labeling ..........................................................................21 
11.3 
Study Drug Administration ....................................................................23 
12. 
STUDY ASSESSMENTS AND PROCEDURES ..............................................24 
12.1 
Baseline Assessments ...............................................................................24 
12.1.1 
Study Day 0 ...............................................................................24 
12.1.2 
Study Day 7 and Study Day 14 .................................................24 
12.1.3 
Study Day 21 .............................................................................24 
13. 
EFFICACY ASSESSMENTS .............................................................................25 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
13.1 
Vitamin D Levels ......................................................................................25 
14. 
ASSESSMENT OF SAFETY ..............................................................................26 
14.1 
Safety Assessments ...................................................................................26 
14.1.1 
Skin Rash or Other Reaction .....................................................26 
14.2 
Reporting of Safety Information ............................................................27 
14.2.1 
Definition of an Adverse Event (AE)........................................27 
14.2.2 
Definition of a Serious Adverse Event (SAE) ..........................27 
14.2.3 
Recording of AEs and SAEs .....................................................28 
14.2.4 
Reporting of SAEs ....................................................................30 
15. 
DATA ANALYSIS AND STATISTICAL CONSIDERATIONS ....................31 
15.1 
Analysis Populations ................................................................................31 
15.2 
Study Endpoints .......................................................................................31 
15.3 
Data Analysis ............................................................................................32 
15.3.1 
Subject Disposition ...................................................................32 
15.3.2 
Demography and Baseline Characteristics................................32 
15.3.3 
Treatment Exposure ..................................................................32 
15.3.4 
Safety Analyses .........................................................................32 
15.3.5 
Analysis of Efficacy ..................................................................32 
15.4 
Determination of Sample Size .................................................................32 
16. 
ETHICS ................................................................................................................33 
16.1 
Ethical Considerations.............................................................................33 
16.2 
Ethical Review Committee ......................................................................33 
16.3 
Informed Consent ....................................................................................33 
17. 
STUDY CONDUCT .............................................................................................34 
17.1 
Regulatory Authorities ............................................................................34 
17.2 
Monitoring Procedures and Quality Assurance....................................34 
17.3 
Curriculum Vitae .....................................................................................34 
17.4 
Investigator Responsibilities ...................................................................34 
17.5 
Emergency Contact with Investigators ..................................................34 
17.6 
Notification of Primary Care Physician .................................................34 
17.7 
Protocol Compliance ................................................................................35 
17.8 
Termination of the Study ........................................................................35 
18. 
ADMINISTRATIVE CONSIDERATIONS ......................................................36 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
18.1 
Protocol Amendments .............................................................................36 
18.2 
Archiving of Records ...............................................................................36 
18.3 
Study Report.............................................................................................36 
19. 
REFERENCES .....................................................................................................37 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
4. 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
Abbreviation 
Definition 
µg 
Microgram(s) 
AE 
Adverse event 
CRF 
Case report form 
g 
Gram(s) 
GCP 
Good Clinical Practice 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IND 
Investigational New Drug 
IRB 
Institutional Review Board 
kg 
Kilogram(s) 
L 
Liter 
MedDRA 
Medical Dictionary for Regulatory Activities 
mg 
Milligram(s) 
mL 
Milliliter 
SAE 
Serious adverse event 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
5. 
INTRODUCTION 
5.1 
Vitamin D Deficiency 
Vitamin D insufficiency is an emerging and compelling public health issue.  Recent studies 
suggest that 19% of the U.S. population is at risk for vitamin D inadequacy 1, although this 
may underestimate the number of people at risk from suboptimal vitamin D levels2. More 
than 40% of U.S. and European elderly men and women living in the community are 
deficient in Vitamin D3, and more than 50% of postmenopausal women taking medication for 
osteoporosis4. Diverse population subgroups are deficient in Vitamin D including Arab 
women and lactose intolerant individuals who consume less vitamin D from fortified 
products5.  In a Boston hospital, 25% of healthy students, physicians, and residents were 
found to be vitamin D–deficient, despite drinking a glass of milk and taking a multivitamin 
daily and eating salmon at least once a week6. By the end of the winter, it has been estimated 
that 42% of 15- to 49-year-old black females throughout the United States have 25-
hydroxyvitamin D levels below 20 ng per milliliter. The implications of this are far-reaching.  
In addition to well-known impacts on bone health, vitamin D status is thought to potentially 
influence a number of common diseases, including cancers, autoimmune disorders, and 
cardiovascular disease. Vitamin D deficiency may increase the risk of injury from falls as 
people age7, while early in life Vitamin D deficiency has been associated with increased risk 
of Type 1 diabetes8,9.  Other patient populations at increased risk for vitamin D insufficiency 
include individuals with fat malabsorption, chronic kidney disease, or obesity10,11,12 .  
  
The Endocrine Society defines vitamin D deficiency as a 25(OH)D of 20 ng/ml (50 
nmol/liter) or below, and vitamin D insufficiency as a 25(OH)D of 21–29 ng/ml (525–725 
nmol/liter)13.  They recommend measuring vitamin D using the serum circulating 25-
hydroxyvitamin D (25(OH)D).  The Institute of Medicine reports that persons are potentially 
at risk for inadequacy at serum 25OHD levels between 12 and 20 ng/mL (30 and 50 
nmol/L)14. In sun-rich environments, normal levels have been estimated to be 54-90 ng/ml15. 
Although it is not known what the safe upper value for 25(OH)D is for avoiding 
hypercalcemia, most studies in children and adults have suggested that the blood levels need 
to be above 150 ng/ml before any concern arises16. 
 
Of all of the forms of vitamin D, cholecalciferol (Vitamin D3) has been viewed as having the 
best overall combination of safety and efficacy, assuming normal kidney and liver function.  
In the US, cholecalciferol has primarily been available as an oral supplement at fairly low 
doses (most commonly 1000 – 2000 IU).   In otherwise healthy patients, a common US 
standard of care for treatment of low serum vitamin D  is to give 50,000 IU Vitamin D orally 
per week for four weeks (total dose 200,000 IU) until levels are brought up into the adequate 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
range, followed by daily 
supplements of 2,000 IU.  This is 
often provided as commercially 
available Vitamin D2, however 
supplementation with Vitamin D3 
appears to be more potent and may 
provide other benefits17.     
 
Despite the widespread availability 
of oral supplementation, the 
frequency of vitamin D deficiency 
remains high.  Some individuals 
have difficulty taking pills, while 
others, especially the elderly, may be 
overwhelmed by their number of oral 
medications.  A need exists for a 
more convenient delivery 
mechanism that enhance access to 
effective long term treatment of vitamin D deficiency. The proposed study aims to 
investigate both safety and efficacy of a topically applied vitamin D that is likely to be more 
effective, convenient, or acceptable for many individuals. 
 
A wide variety of dosing schedules for Vitamin D have been documented18. Recently Vieth’s 
team conducted a study with oral cholecalciferol in 48 elderly women (81 ± 8 years)19.  The 
study tested oral cholecalciferol (D3) up to 45,000 IU once every 28 days.  The study showed 
that a single oral dose of 45,000 IU dissolved in alcohol generally produced a rise in serum 
vitamin D of about 10 ng/ml.  A second oral dose 28 days later further increased the serum 
vitamin D level by about another 10 ng/ml.  Thus, the total increase after two oral doses 
(90,000 IU) was about 20 ng/ml.   Although following oral dosing, rises in serum Vit D can 
be seen up to one month following administration, Vieth’s study demonstrated that most of 
the rise (>60%) in Vitamin D occurs within 7 days. (Figure 1). No one receiving oral 
monthly dosing of 45,000 IU Vitamin D3 developed hypercalcemia.  
 
Vitamin D Product Development 
Innovative Pharmacy Services, Inc. has developed a vitamin D3 topical gel using ingredients 
that are identified in the USP/NF and/or designated in the list of Inactive Ingredients in FDA 
Approved Drugs.  The ingredients in the topical gel are well known in the industry and have 
references in the pharmaceutical literature and some have available FDA drug master file 
information.  A single applicator contains 0.75 g of gel, containing 150,000 international 
units of vitamin D3 (cholecalciferol).  One dose will consist of two applicators.  One 
applicator will be applied to each of the subject’s upper arm for a total dose of 300,000 units.   
 
Figure 1. Absorption kinetics of oral Vitamin D3. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
5.2 
Study Rationale and Selection of Dose 
5.2.1 
Rationale for Study 
The purpose of this investigation is to demonstrate that a novel topical vitamin D3 
(cholecalciferol) gel is bioavailable to the systemic circulation and in sufficient quantities to 
treat hypovitaminosis D. In Phase Ia, healthy volunteers will be dosed with a topical gel to 
determine increases of serum 25-OH vitamin D that can be expected from three doses.  The 
application will take place in the winter months in Minnesota.  Although ambient levels of 
light are low at that time, subjects will be asked to avoid attending tanning booths, and to 
wear a comfortable shirt that covers their upper arms at least to the elbow if they choose to 
go outside during daylight hours.  
5.2.2 
Rationale for Selected Dose 
A wide variation in Vitamin D dosing has been used in clinical practice12.  If we assume that 
the topical gel vitamin D dosing has no greater bioavailability than intramuscularly injected 
dosing, then the maximal effect of 100,000 IU of Vitamin D3 applied topically would be 
expected to reflect that of a single 100,000 IU injected dose19.  The supplementation protocol 
most often used in France to correct vitamin D levels of  ≥ 10 ng/ml and <20 ng/ml is three 
doses of 100,000 IU of Vitamin D3 given intramuscularly one week apart20. For values ≤ 
10ng/ml four doses of 100,000 IU are administered.  An observational study of this common 
French protocol by Rouillon evaluated 204 patients receiving 3-4 doses of 100,000 IU 
Vitamin D3 weekly for vitamin deficiency (Groups 1 and 2)21.  Rouillon found the protocol 
to be safe with no episodes of hypercalcemia. Two patients from the total of 257 studied 
developed Vitamin D levels over 90 ng/ml (one 92 mg/ml and one 94 ng/ml), with both 
resolving within one month.   .  
     
The proposed study will use a topical application of Vitamin D which is unlikely to be as 
completely absorbed as intramuscular injection.  Furthermore, topical application of Vitamin 
D provides important additional physiologic mechanisms that reduce the potential for 
possible hypervitaminosis. Upon exposure to UVB radiation, previtamin D3 is synthesized 
from 7-DHC in the stratum basale and stratum spinosum layers of the epidermis. Previtamin 
D3 thermoisomerizes to vitamin D3 with heat or sunlight exposure at the ultraviolet region of 
the solar spectrum around 316-330 nm 22.  Vitamin D3 is then available for binding with a 
vitamin D binding protein (DBP) and transported into the circulation23. In the skin, the DBP 
acts as a regulatory mechanism preventing over absorption of Vitamin D into the circulation 
by providing a threshold when the binding protein has been fully occupied.  Upon exposure 
to sunlight, vitamin D3 itself is rapidly degraded to other photoproducts, including 5,6-trans-
vitamin D3, suprasterol I, and suprasterol II 24.  Although winter sunlight is not strong 
enough to produce vitamin D3 in the skin, it is strong enough to promote the 
photodestruction of existing vitamin D 24.  Neither binding with DBP nor other serum 
components prevent photodestruction.).  
 
In the process of normal physiologic production, previtamin D3 can isomerize in a reversible 
reaction to form the photoproducts, tachysterol-3 or lumisterol-3.  These isomers do not 
enhance intestinal calcium absorption and do not significantly enter the circulation.  The 
inactive metabolic products are sloughed off when skin cells are naturally shed. Removal of 
previtamin D3 provides a mechanism to prevent vitamin D toxicity from prolonged sun 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
exposure25, and may further increase the potential safety of topical Vitamin D3 
administration.  The Mayo Clinic reports that Vitamin D is ‘likely’ safe when applied to the 
skin12.   
 
The effectiveness of topical absorption for the proposed product is unknown, however topical 
absorption is substantially less effective than the intramuscular injection of Vitamin D3.  In 
the first three candidates, 300,000 units applied over three weeks resulted in an insignificant 
average change in Vitamin D level, as follows.      
Baseline 
Week 1 
Week 2 
Week 3 
Av 
change 
10.2 
9.4 
10.4 
12.1 
1.2 
 
 
Oral administration of 50,000 IU should produce a 10 ng/ml increase in total serum Vitamin 
D 13.   The topical application os 300,000 IU, therefore, was substantially less effective than 
one 50,000 IU oral dose.  .  The study will use three doses of 300,000 IU applied one week 
apart with safety and outcome measures after each.  This is hoped to provide a greater 
difference in vitamin D level than that seen in one oral 50,000IU supplement, but should 
produce less change than the typical US replacement dosing of three oral supplements,  If the 
change in total serum Vitamin D level after the topical application of 900,000 IU in the first 
10 subjects is consistently less than 10 mg/ml then the study will be discontinued.   
 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
6. 
STUDY OBJECTIVES 
The primary purpose of the study is to assess the change in serum 25-OH vitamin D 
concentrations following 3 doses of 300,000 IU of a novel vitamin D3 gel.  The secondary 
objective is to assess the safety (based on any skin rash or other skin reactions, as well as 
calcium and parathyroid hormone levels) following treatment. 
. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
7. 
INVESTIGATIONAL PLAN 
7.1 
Overall Study Design 
20 healthy adults with known vitamin D deficiency (defined as serum 25-OH vitamin D 
concentration < 20 ng/ml) will receive 3 doses of 300,000 IU of topical vitamin D once per 
week for three weeks.  Each dose will be made up of two applications of 150,000 IU 
cholecalciferol in .75 ml of gel applied consecutively to the upper arms (one 150,000 IU 
application per arm).  
 
Blood will be drawn at baseline prior to the first dose (Day 0) and on Days 7, 14, and 21.    
Total serum 25-OH vitamin D will be assayed at each draw. Parathyroid hormone level will 
be assayed at baseline and on day 21.     
7.2 
Study Duration 
The maximum duration of treatment is 14 days (study Days 0, 7 and 14). Subjects will be 
followed for change in vitamin D blood level and for safety until Study Day 21.  It is 
anticipated that the study will last 4 months to enroll and complete dosing and assessment of 
20 patients. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
8. 
STUDY POPULATION 
8.1 
Inclusion Criteria 
A subject must meet all of the following inclusion criteria to be eligible for enrollment in this 
study: 
1. Known vitamin D deficiency based on blood work obtained prior to study consent 
defined as serum 25-hydroxyvitamin D less than or equal to 20 ng/mL within 30 days 
prior to study entry. 
2. Adult, age 18 to 85 years, male or female 
3. If on oral vitamin D therapy, have been on a stable dose for the previous 90 days. 
8.2 
Exclusion Criteria 
Subjects are excluded from this study if any of the following conditions are met: 
1. History of acute or chronic liver disease with elevated liver function tests, chronic 
kidney disease (stage 3 or greater, eGFR <60), uncontrolled thyroid disease (elevated 
thyroid function tests), primary or secondary hyperparathyroidism, hypercalcemia, or 
multiple endocrine neoplasia. 
 
2. Hypercalcemia defined as elevated corrected serum calcium (>10.2 mg/dl) measured 
within 3 months prior to study. 
 
3. eGFR  <60 mL/min within 3 months prior to study. 
4. Active cancers 
5. Women who are pregnant or breastfeeding  
6. Individuals who are unable to give informed consent 
7. Individuals with psoriasis, active eczema or other active skin disease, or who are 
currently receiving treatments or medications to control skin disease  
8. Individuals who do not agree to refrain from using tanning beds for the duration of 
the study. 
9. Individuals who do not agree to avoid submerging the gel site in water for 8 hours 
after gel application. 
10. 10. Family members of the study faculty or staff.  
8.3 
Criteria for Withdrawal 
A subject has the right to withdraw fully or partially from the study at any time and for any 
reason without prejudice to his or her future medical care by the physician or at the 
institution.  
Withdrawal of full consent for a study means that the subject does not wish to receive further 
investigational treatment and does not wish to or is unable to continue further in study 
participation.  Subject data up to withdrawal of consent will be included in the analysis of the 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
study.  The Investigator will discuss with the subject appropriate procedures for withdrawal 
from the study.  Withdrawal of partial consent from a study means that the subject does not 
wish to take investigational product or other protocol-required therapy any longer but is still 
willing to collaborate in providing further data by continuing on study.  Should a subject 
request or decide to withdraw from the study, all efforts will be made to complete and report 
the observations as thoroughly as possible up to the date of withdrawal.  All information 
should be reported on the applicable case report form (CRF).  Reasons for removal from 
protocol-required treatment or observation might include: 
 partial/full withdrawal of consent 
 administrative decision by the Sponsor-Investigator 
 pregnancy 
 ineligibility 
 significant protocol deviation 
 subject noncompliance 
 adverse event, including rash or other skin reaction 
 Death 
8.4 
Replacement of Subjects 
Subjects who withdraw or are removed from the study prior having their Day 7 blood draw 
for a reason other than a drug-related adverse event will be replaced.   

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
9. 
TREATMENT OF SUBJECTS 
9.1 
Treatment Assignment 
This is an open-label, nonrandomized study in which all subjects will receive topical vitamin 
D gel.  
9.2 
Treatment Administration 
Subjects will be provided with written and verbal instructions about Vitamin D deficiency 
during their baseline/Day 1 visit.  A study nurse or physician will supervise each treatment 
application to ensure that the gel is applied correctly. Prior to application of the gel, 2 strips 
of paper tape will be applied to the subject’s upper arms. One strip will be applied 
approximately 2 inches below the subject’s shoulder and the second strip will be applied 
approximately 2 inches above the subject’s elbow.  The entire contents of one syringe (0.75 
mL of gel) will be applied onto their arm in a straight line, from approximately 1 inch below 
the first strip of paper tape to approximately 1 inch above the second strip of paper tape.  The 
subject will be instructed to gently rub the gel into their upper arm with their fingertips, while 
staying between the two pieces of paper tape, for approximately 2 minutes or until the gel has 
been absorbed.  After completing the application process on both arms, the subject will wash 
his/her hands thoroughly. 
A complete description of the administration process can be found in Appendix 1. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
10. 
STUDY DRUG MATERIALS AND MANAGEMENT 
10.1 
Description of Study Drug 
A novel topical vitamin D3 (cholecalciferol) gel has been developed and is being provided by 
Innovative Pharmacy Solutions, Inc. The ingredients in the gel are well-known in the 
industry and have references in the pharmaceutical literature and some have available FDA 
drug master file information. The formulation for the topical vitamin D gel is: 
 
Gel Formula 
Vitamin D3 1,000,000 IU/gm      
 
 
 
 
20.0 gm 
Phosol 53 MCT (lipoid)   
 
 
 
 
 
20 gm 
Cholesterol USP-NF   
 
 
 
 
 
 
0.8 gm 
Isopropyl myristate USP-NF  
 
 
 
 
 
5 gm 
Poloxamer 407 NF  
 
 
 
 
 
 
 
7.5 gm 
Purified Water USP 
 
 
 
 
 
 
 
46.5 gm 
Methylparaben USP-NF  
 
 
 
 
 
0.18 gm 
Propylparaben  USP-NF  
 
 
 
 
 
0.02 gm 
The previous anhydrous formula was changed to a hydrous base to provide a less greasy 
product.  The new formula is a simple wash off base that provides phospholipids that are able 
to self-associate into micelles and liposomes.  
 
10.2 
Packaging and Labeling 
The topical vitamin D gel is provided in a syringe as a single application of 0.75g of gel, 
containing 150,000 IU of vitamin D3 (cholecalciferol) that will be applied to each of the 
subject’s upper arms, totaling 300,000 IU per dose. 
Labeling of study drug will comply with regulatory requirements as follows: 
 
 
 
 
 
II.G. Labeling 
II.G. 1. Anhydrous Base 
 
 
 
 
 
 
 
 
 
 
Hydrous Base 
  
Date Made 04-10-2014 
Exp 07-10-14  
Compounded Product 
Lot # 20140410  
IND# 110937  
KEEP REFRIGERATED 
Caution New Drug – 
Limited by Federal Law to 
Investigational Use 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
 
 
 
 
 
II.G. 2. Drug Product 
1. The label for the syringe with the final product will include the product name, strength 
and quantity, mixing date, beyond use date, name of the pharmacy compounding the 
product (Innovative Pharmacy Solutions), IND number, RPh name that compounded and 
dispensed and auxiliary labels “Keep Refrigerated” and “CAUTION New Drug – 
Limited by Federal law to Investigational Use.” 
 
 
 
 
 
 
 
 
 
 
 
 
2. The ziplock bag label will contain the compounding pharmacy name (Innovative 
Pharmacy Solutions), address, phone number and after hours phone number, IND 
number, “Keep Refrigerated” and “CAUTION New Drug – Limited by Federal law to 
Investigational Use.” 
 
 
 
 
 
 
 
 
 
 
 
3. The 1 mL syringe labels will contain the product name and “For Left Arm 
administration” or “For Right Arm administration”, IND number, and “CAUTION New 
Drug – Limited by Federal law to Investigational Use.” 
 
 
 
Innovative Pharmacy Solutions 
IND# 110937 
Vitamin D3 in Hydrous Base 200,000 IU/mL 
Quantity: 0.25ml 50,000 IU VitaminD3 
Mix Date: 7.28.2015      Beyond Use Date: 7.28.2015 
RPh name:                     Toll Free Phone 1-800-844-9845 
KEEP REFRIGERATED 
  CAUTION New Drug - Limited by Federal Law to Investigational Use 
Innovative Pharmacy Solutions 
IND# 110937 
323 Jackson Avenue Elk River, MN 44330 
Minnesota Pharmacy License # 263570 
Pharmacy Phone 763-421-4788  
Steve Cell 612-270-5504 Toll Free Phone  1-800-805-9845 
KEEP REFRIGERATED 
CAUTION New Drug - Limited by Federal Law to Investigational Use 
For Left Arm Administration 
50,000 IU Vitamin D3 in Anhydrous Base 0.25 ml 
IND# 110937   
CAUTION New Drug - Limited by Federal Law to Investigational Use 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
 
 
 
 
 
 
 
 
 
 
 
 
 
10.3 
Study Drug Administration 
A study nurse or physician will supervise each treatment application to ensure that the gel is 
applied correctly.  Prior to application of the gel, 2 strips of paper tape will be applied to the 
subject’s upper arms. One strip will be applied approximately 2 inches below the subject’s 
shoulder and the second strip will be applied approximately 2 inches above the subject’s 
elbow.  The entire contents of one syringe (0.75 mL of gel) will be applied onto each arm in 
a straight line, from approximately 1 inch below the first strip of paper tape to approximately 
1 inch above the second strip of paper tape.  The subject will be instructed to gently rub the 
gel into their upper arm with their fingertips, while staying between the two pieces of paper 
tape, for approximately 2 minutes or until the gel has been absorbed.  After completing the 
application process on both arms, the subject will wash his/her hands thoroughly.  
 
Handling and Storage of Study Drug 
Specific instructions for preparing and administering the topical vitamin D gel are provided 
in Appendix 1. 
For Right Arm Administration 
50,000 IU Vitamin D3 in Anhydrous Base 0.25 ml 
IND# 110937 
CAUTION New Drug - Limited by Federal Law to Investigational Use 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
11. 
STUDY ASSESSMENTS AND PROCEDURES 
The schedule of study assessments and procedures is shown in Table 1.   
11.1 
Baseline Assessments 
A chart review of subjects with known vitamin D deficiency will be performed prior to Day 0 
in order to assess for potential study eligibility. Informed consent must be obtained before 
any study procedures are performed.   
11.1.1 
Study Day 0 
The following procedures should be completed on Day 0: 
 informed consent  
 assessment of height, weight and skin to assess for study eligibility 
o Anthropometric Assessment 
 Patients are measured for height (cm) and weight (kg) following 
standardized procedures as outlined in the Anthropometric Standardization 
Reference Manual (Lohman et al., 1991).    Body weight is measured with 
a stand-on digital scale and height is measured by a stadiometer.  Height is 
measured to the nearest 0.1 cm and weight to the nearest 0.1 kg.   
o Recording of Fitzpatrick skin-type score.  
 Blood draw for vitamin D level, parathyroid hormone level, calcium, and albumin. 
 Application of the topical vitamin D gel, including instructions for dosing at home 
11.1.2 
Study Day 7 and Study Day 14 
 Evaluation of skin site where gel applied.  If the subject has any skin rash or other 
reaction, he/she should contact study personnel as soon as possible for photographs 
and an assessment/diagnosis 
o Photographs of application sites 
 A digital photograph may be taken of the gel application site following 
application to record any signs of irritation or reaction. Identifying 
marks on or near the application site (i.e. tattoos, birthmarks, scars 
etc.) should be avoided when a photograph is taken. Digital 
photographs will be uploaded to the REDcap database and stored on 
the secure server.  
 Blood draw for vitamin D level and calcium 
 Application of the topical vitamin D gel 
11.1.3 
Study Day 21 
 Assessment of skin for rash or any other reaction 
 Blood draw for vitamin D, parathyroid hormone level, and calcium. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
12. 
EFFICACY ASSESSMENTS 
Efficacy assessments should be performed according to the Schedule of Events (Table 1).   
12.1 
Vitamin D Levels 
Assessments of serum 25-OH vitamin D levels (D2 and D3) will be done and on Day 0, 7, 
14, and 21. Venous blood draws will be performed by a trained University of Minnesota 
Physicians (UMP) technician and delivered to HealthEast laboratory for analysis.    

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
13. 
ASSESSMENT OF SAFETY 
13.1 
Safety Assessments 
13.1.1 
Skin Rash or Other Reaction 
An assessment of each subject’s skin will be performed on Days 7, 14 and 21 using the 
following scoring system for topical irritation developed per FDA guidance: 
Basic Score:  Description: 
 0  No visible reaction 
 
 0.5   Doubtful or negligible erythema reaction 
 
 1.0  
Mild or just perceptible macular erythema reaction in a speckled/follicular, 
patchy or confluent pattern (slight pinking) 
 
 2.0 
Moderate erythema reaction in a confluent pattern (definite redness) 
 
 3.0  
Strong or brisk erythema reaction that may spread beyond the test site 
 
Supplemental Scores: 
Description: Label: 
 
0.5  
 
Edema  
E 
 
0.5  
 
Papules 
P 
 
0.5  
 
Vesicles 
V 
 
0.5  
 
Bullae  
B 
Supplemental scores will be added to the basic scores if the associated symptoms are present. The 
final score will be the sum of the basic and supplemental scores.  
As part of the general safety assessment, dermal irritation will be evaluated by the 
Investigator or designated research staff using the outlined scale.  The likelihood of allergic 
reaction not manifested as a dermal irritation is extremely low.  Given that all the ingredients 
used to make the topical vitamin D gel are considered GRAS, we do not anticipate non-local 
allergic reactions.  In the event that a non-dermal allergic reaction occurs, standard medical 
procedures utilized for such situations will be followed.   
 
If a subject develops a dermal response of 1.5 up to 2.5, he or she will be given the option to 
permanently discontinue the study treatment and withdraw from the study, or to allow 
sufficient time for the rash to resolve before he or she reinitiates the phase of the study he or 
she was in at the time that the rash developed.  If a participant develops a dermal response of 
2.5 or greater, he or she will be required to permanently discontinue the study treatment and 
he or she will be withdrawn from the study.  In both cases, the study participant will be 
encouraged to seek medical care from his or her primary care physician. In addition, we will 
ask subjects who decide to withdraw from the study to return for their next scheduled blood 
draw, as per protocol.  We will also follow up with any study participant who withdraws 
from the study due to AE occurrence through telephone contact at 1 week intervals until the 
AE resolves. 
 
All other adverse events will be assessed by self-report from subjects.  
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
We will initially administer 100,000 units topically to subjects, and check serum Vitamin D 
and calcium levels one week later.  Dosing will be discontinued if a patient achieves a 
Vitamin D level of greater than normal (> 90 ng/ml) or if serum calcium rises above normal 
(> 10.2 mg/dl). Additional doses of 100,000 units will be administered on days 7 and 14. The 
outcome of interest will be the effect on total serum Vitamin D at 21 days. Although a 
change in calcium is unlikely, calcium will be monitored for safety.   
 
Vitamin D may alter glucose values and improve glucose control in individuals with 
diabetes26. Although individuals with diabetes will not be excluded, we will instruct any 
subjects with diabetes to more closely monitor glucose values during the course of the study.  
 
Parathyroid hormone will be measured at baseline and on Day 21 to determine if vitamin D 
correction changes the level of this metabolic calcium regulatory hormone.    
 
 
13.2 
Reporting of Safety Information 
13.2.1 
Definition of an Adverse Event (AE) 
Any untoward medical occurrence in a clinical investigation subject, temporarily associated 
with the use of a medicinal product, whether or not considered related to the medicinal 
product. 
Examples of an AE include: 
 Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition 
 A new condition detected or diagnosed after investigational product administration 
even though it may have been present prior to the start of the study 
 Signs, symptoms, or the clinical sequelae of a suspected interaction 
 Signs, symptoms, or the clinical sequelae of a suspected overdose of either 
investigational product or a concurrent medication (overdose per se should not be 
reported as an AE/SAE) 
Examples of an AE do not include: 
 Medical or surgical procedure (e.g., endoscopy, appendectomy); the condition that 
leads to the procedure is an AE 
 Situations where an untoward medical occurrence did not occur (social and/or 
convenience admission to hospital, such as for PK sampling convenience) 
In this study, AEs may include pre- or post-treatment events that occur as a result of 
protocol-mandated procedures (i.e., invasive procedures, modification of a subject’s previous 
therapeutic regimen). 
13.2.2 
Definition of a Serious Adverse Event (SAE) 
An SAE is any untoward medical occurrence that, at any dose: 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
a. results in death 
b. is life-threatening 
Note: The term ‘life-threatening’ in the definition of ‘serious’ refers to an event in which 
the subject was at risk of death at the time of the event.  It does not refer to an event, 
which hypothetically might have caused death if it were more severe. 
c. requires hospitalization or prolongation of an existing hospitalization 
Note: In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for observation 
and/or treatment that would not have been appropriate in the physician’s office or out-
subject setting.  Complications that occur during hospitalization are AEs.  If a 
complication prolongs hospitalization or fulfills any other serious criteria, the event is 
serious.  When in doubt as to whether ‘hospitalization’ occurred or was necessary, the 
AE should be considered serious. 
Hospitalization for elective treatment of a pre-existing condition that did not worsen from 
baseline is not considered an AE. 
d. results in disability/incapacity 
Note: The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.  This definition is not intended to include experiences of relatively 
minor medical significance such as uncomplicated headache, nausea, vomiting, diarrhea, 
influenza, and accidental trauma (e.g., sprained ankle) which may interfere or prevent 
everyday life functions, but do not constitute a substantial disruption. 
e. is a congenital abnormality / birth defect 
f. any event deemed by the Investigator as being a significant medical event. 
Medical and scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be immediately 
life-threatening or result in death or hospitalization, but may jeopardize the subject or may 
require medical or surgical intervention to prevent one of the other outcomes listed in the 
above definition.  These should also be considered serious.  Examples of such events are 
invasive or malignant cancers, intensive treatment in an emergency room or at home for 
allergic bronchospasm, blood dyscrasias or convulsions that do not result in hospitalization, 
or development of drug dependency or abuse.   
13.2.3 
Recording of AEs and SAEs 
When an AE/SAE occurs, it is the responsibility of the Investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostic reports) relative to 
the event. The Investigator will then record all relevant information regarding an AE/SAE 
into the CRF.   
For each AE, start and stop dates, action taken, outcome, intensity (see Section 13.2.3.1) and 
relationship to study product (causality) (see Section 13.2.3.2) must be documented.  If an 
AE changes in frequency or intensity during a study, a new entry of the event must be made 
in the CRF. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
The Investigator will attempt to establish a diagnosis of the event based on signs, symptoms, 
and/or other clinical information.  In the absence of a diagnosis, the individual 
signs/symptoms should be documented. 
All details of any treatments initiated because of the AE should be recorded in the subject’s 
notes and the CRF. 
All AEs and SAEs will be followed until resolution, until the condition stabilizes, until the 
event is otherwise explained, or until the subject is lost to follow-up.  Once resolved, the 
appropriate AE/SAE CRF page(s) will be updated.  The Investigator will ensure that follow-
up includes any supplemental investigations as may be indicated to elucidate the nature 
and/or causality of the AE or SAE.  This may include additional laboratory tests or 
investigations, histopathological examinations, or consultation with other health care 
professionals.  
13.2.3.1 
Assessment of Intensity 
The Investigator will make an assessment of intensity for each AE and SAE reported during 
the study.  The assessment will be based on the Investigator’s clinical judgment.  Adverse 
events from the first dose of topical vitamin D gel will be assesed based on this general 
guideline:  
 Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; 
intervention not indicated 
 Moderate; minimal, local or non-invasive intervention indicated; limiting age-
appropriate instrumental activities of daily  
 Severe or medically significant but not immediately life-threatening; hospitalization 
or prolongation of hospitalization indicated; disabling; limiting self care activities of 
daily living 
 Life-threatening consequences; urgent intervention indicated 
 Death related to AE  
Severity should not be confused with an SAE.  Severity is a category utilized for rating the 
intensity of an event; and both AEs and SAEs can be assessed as severe.  An event is defined 
as “serious” when it meets one of the pre-defined outcomes as described in Section 13.2.2 
“Definition of an SAE.” 
13.2.3.2 
Assessment of Causality 
The Investigator is obligated to assess the relationship between investigational product and 
the occurrence of each AE/SAE.  The Investigator will use clinical judgment to determine the 
relationship.  Alternative causes, such as natural history of the underlying diseases, 
concomitant therapy, other risk factors, and the temporal relationship of the event to the 
investigational product will be considered and investigated.   
The causal relationship to the study product assessed by the Investigator (or medically 
qualified delegate) should be assessed using the following classifications: 
Not Related In the Investigator’s opinion, there is not a causal relationship between the 
study product and the AE. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
Unlikely 
The temporal association between the AE and study product is such that the 
study product is not likely to have any reasonable association with the AE. 
Possible 
The AE could have been caused by the study subject’s clinical state or the 
study product. 
Probable 
The AE follows a reasonable temporal sequence from the time of study 
product administration, abates upon discontinuation of the study product and 
cannot be reasonably explained by the known characteristics of the study 
subject’s clinical state. 
Definitely 
The AE follows a reasonable temporal sequence from the time of study 
product administration or reappears when study product is reintroduced. 
The causality assessment is one of the criteria used when determining regulatory reporting 
requirements. 
13.2.3.3 
Assessment of Expectedness 
Expected 
An adverse reaction, the nature or severity of which is consistent with the 
applicable product information (e.g., Investigators’ Brochure for an 
unapproved medicinal product). 
Unexpected An adverse reaction, the nature or severity of which is not consistent with 
information in the relevant source document (e.g., Investigators’ Brochure for 
an unapproved medicinal product). 
13.2.4 
Reporting of SAEs 
Once a Sponsor-Investigator becomes aware that an SAE has occurred in a study subject, 
he/she will assess the event and determine if it meets regulatory reporting requirements. The 
Investigator must also notify the reviewing Investigational Review Board (IRB) and site 
governance office (if required) of any SAEs according the guidelines of the IRB. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
14. 
DATA ANALYSIS AND STATISTICAL CONSIDERATIONS 
Data for this study will be entered into a REDCap database, which uses a MySQL database 
via a secure web interface with data checks used during data entry to ensure data quality. 
REDCap includes a complete suite of features to support HIPAA compliance, including a full 
audit trail, user-based privileges, and integration with the institutional LDAP server. The 
MySQL database and the web server will both be housed on secure servers operated by the 
University of Minnesota Academic Health Center’s Information Systems group (AHC-IS). 
The servers are in a physically secure location on campus and are backed up nightly, with the 
backups stored in accordance with the AHC-IS retention schedule of daily, weekly, and 
monthly tapes retained for 1 month, 3 months, and 6 months, respectively. Weekly backup 
tapes are stored offsite. The AHC-IS servers provide a stable, secure, well-maintained, and 
high-capacity data storage environment, and both REDCap and MySQL are widely-used, 
powerful, reliable, well-supported systems. Access to the study's data in REDCap will be 
restricted to the members of the study team by username and password. 
The general analytical approach for all endpoints will be descriptive in nature. No formal 
statistical hypothesis testing will be conducted. No p-values will be presented due to the 
small sample size of this study. Data analyses will be provided for all study subjects 
combined wherever appropriate.   
For continuous variables, summary statistics will include number of subjects, mean, median, 
standard deviation, minimum, and maximum.  For categorical variables, descriptive statistics 
will include number of subjects and percentages.  
In regards to handling of missing data, there will be no imputation of missing values. 
14.1 
Analysis Populations 
Safety Population:  all subjects who receive at least 1 dose of study drug and who have at 
least one post-dosing safety evaluation.  All safety endpoints, baseline characteristics, and 
demographic data will be summarized using the Safety Population. 
Efficacy Evaluable Population:  all subjects who receive all 3 doses of study drug and have a 
follow up blood draw for vitamin D levels, or discontinue prior to completing dosing due to a 
drug-related adverse event.  Vitamin D level data will be analyzed using the Efficacy 
Evaluable Population.   
14.2 
Study Endpoints 
The primary endpoint is to establish the change in vitamin D blood levels following 3 doses of 
treatment with vitamin D gel. 
Secondary study endpoints include assessment of safety, including change in parathyroid hormone 
level and the number of subjects who have any skin rash and/or other skin reaction following 
treatment with the vitamin D gel. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
14.3 
Data Analysis 
14.3.1 
Subject Disposition 
The number and percentage of subjects entering and discontinuing the clinical study will be 
presented.  The reasons for discontinuation will also be summarized. 
14.3.2 
Demography and Baseline Characteristics 
Demographic and baseline characteristics will be descriptively summarized using the Safety 
Population.  Quantitative and/or categorical summaries will be presented for demographics 
and other baseline characteristics.   
14.3.3 
Treatment Exposure 
Treatment exposure will be summarized.  Measures of extent of exposure include the total 
number of doses per subject, cumulative dose per subject, and compliance (percent of target 
dose received per cycle).  The reason(s) for dose(s) missed will also be summarized. 
14.3.4 
Safety Analyses 
Safety parameters will be listed and summarized using standard descriptive statistics.   
Adverse event terms will be coded using the Medical Dictionary for Regulatory Activities 
(MedDRA) and will be summarized by dose.  Incidence of AEs occurring during the study 
will be summarized by system organ class (SOC) and preferred term.  Adverse events will 
also be summarized by causality and grade.  Serious adverse events and AEs leading to 
discontinuation of study treatment will be listed separately.   
Additional analyses will be performed if warranted upon review of the data. 
14.3.5 
Analysis of Efficacy 
The primary outcome will be the change in total Vitamin D following the topical application 
of 100,000IU Vitamin D3 weekly for three weeks.    
14.4 
Determination of Sample Size 
A change from baseline of 10 µg/L in total 25-OH Vitamin D levels in subjects with Vitamin 
D deficiency is thought to be a clinically relevant improvement. Assuming that the standard 
deviation for pre and post measures of total 25-OH Vitamin D is 4 μg/L, then the standard 
deviation of the difference is 5.6 μg/L so 10 subjects should be adequate to show an effect 
size of 6.4 μg/L with alpha of .05 and a power of 90%.  However the standard deviation of 
the second measure is unknown since absorption may vary by individual.  Assuming the 
standard deviation of the second measure increases to 12, then 20 subjects would be needed 
to detect an effect size of 10 μg/L.  In order to be most conservative, 20 subjects will be 
treated in this proof-of-concept study. 
 
 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
ETHICS 
14.5 
Ethical Considerations 
This study will be carried out according to the Declaration of Helsinki, the National 
Statement on Ethical Conduct in Human Research (2007) and the Notes for Guidance on 
Good Clinical Practice as adopted by the Australian Therapeutic Goods Administration 
(TGA; 2000) (CPMP/ICH/135/95) and the ICH GCP Guidelines27.  
14.6 
Ethical Review Committee 
The Protocol will be submitted for review to the University of Minnesota Investigational 
Review Board (IRB), and written approval will be obtained from both the IRB and 
Governance Office (as required), before subjects are recruited and enrolled.  The 
Investigators will receive all the documentation needed for submitting the protocol to the 
IRB and/or Governance Office.  A copy of the respective approval letters will be obtained 
before starting the study.  If approval is suspended or terminated by the IRB or Governance 
Office, the Sponsor-Investigator will have responsibility for notifying the Food and Drug 
Administration. 
It is the responsibility of the Investigator to report study progress to the IRB or Governance 
Office as required. 
The Sponsor-Investigator, or his/her nominee, will be responsible for reporting any SAEs to 
the IRB as required. 
14.7 
Informed Consent 
Before recruitment and enrollment into the study, each prospective subject will be given a 
full explanation of the nature and purposes of the study.  Once the essential study information 
has been provided, and the Investigator is assured that the volunteer understands the 
implications of participating in the study, the subjects will be asked to give consent to 
participate in the study by signing the Informed Consent Form (ICF)/Patient information 
sheet.  The consent form will be signed and dated by the appropriate parties.  A notation that 
written informed consent has been obtained will be made on the subject’s medical record and 
CRF.  The completed consent form will be retained by the Investigator.  A copy of the 
completed ICF will be provided by the Investigator to the subject. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
15. 
STUDY CONDUCT 
15.1 
Regulatory Authorities 
This study will be conducted under an Investigational New Drug (IND 110937). 
15.2 
Monitoring Procedures and Quality Assurance 
The organization, monitoring, supply of study materials and quality assurance of the present 
clinical study is the responsibility of Innovative Pharmacy Solutions or its designee. 
Monitoring may be conducted to ensure compliance with the protocol, ICH GCP and 
applicable regulatory requirements.  
In order to ensure the accuracy of data, direct access to source documents by the 
representatives of Innovative Pharmacy Solutions, Inc. and regulatory authorities is 
mandatory.   
The Investigator is required to submit to the Reviewing IRB, annual (or more frequent if 
requested) reports of the study.   
15.3 
Curriculum Vitae 
A current, signed and dated Curriculum Vitae for all study staff will be provided and 
maintained in the files at Innovative Pharmacy Solutions, Inc. (or designee).  A Staff 
Signature List will be maintained at each Investigator's site.  
15.4 
Investigator Responsibilities 
Except where the Principal Investigator’s signature is specifically required, it is understood 
that the term ‘Investigator’ as used in this Protocol and on the CRFs refers to the Principal 
Investigator or an appropriately qualified member of the staff that the Principal Investigator 
designates to perform specified duties of the Protocol.  The Principal Investigator is 
ultimately responsible for the conduct of all aspects of the study.  
Each Investigator will comply with the local regulations regarding clinical trials and the 
Investigator responsibilities outlined in the ICH GCP guidelines. 
15.5 
Emergency Contact with Investigators 
All subjects will be provided with a Participant Emergency Contact Card with contact details 
of whom to contact in the case of an emergency. 
15.6 
Notification of Primary Care Physician 
With the consent of the subject, it is the Investigator’s responsibility to notify the primary 
care physician of the subjects’ involvement in the study, provided that such a physician can 
be identified for the subject.  A letter will be sent to the physician stating the nature of the 
study, treatments, expected benefits or AEs and concomitant drugs to be avoided.  A copy 
shall be retained by the study site. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
15.7 
Protocol Compliance 
The instructions and procedures specified in this protocol require diligent attention to their 
execution.  Any subject treated in a manner that deviates from the protocol, or who is 
enrolled into the study but is not qualified according to the Protocol may be ineligible for 
analysis and thereby compromise the study. 
Only when an emergency occurs that requires a departure from the Protocol for an individual 
will there be such a departure.  The nature and reasons for the Protocol violation/deviation 
shall be recorded in the CRF.  
The Investigator and designees will comply with all applicable federal, state and local laws. 
15.8 
Termination of the Study 
The Sponsor-Investigator reserves the right to terminate the clinical study at any time.  
Reasons for termination may include, but are not limited to, the following: 
 the incidence or severity of AEs in this study 
 serious or persistent noncompliance by the Investigator with the protocol, clinical 
research agreement, or applicable regulatory guidelines in conducting the study 
 IRB decision to terminate or suspend approval for the investigation or the Investigator 
 Investigator request to withdraw from participation 
In addition the Sponsor-Investigator reserves the right to discontinue the study prior to 
inclusion of the intended number of subjects, but intends only to exercise this right for valid 
scientific or administrative reasons.  
After such a decision, the Investigator must contact all participating subjects promptly, and 
written notification must be sent to the Reviewing IRB and relevant Governance Offices (as 
required).   

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
16. 
ADMINISTRATIVE CONSIDERATIONS 
16.1 
Protocol Amendments 
Protocol modifications that impact on subject safety or the validity of the study will be 
approved by the IRB. If a Protocol amendment requires changes to the Informed Consent 
Form, the revised Informed Consent Form, prepared by the Investigator, must be approved 
by the IRB and site governance offices (as required). 
Once the final Protocol has been issued and approved by the Sponsor-Investigator and the 
authorized signatories, it shall not be informally altered.  Protocol amendments are alterations 
to a legal document and have the same legal status.  Therefore, they must pass through 
appropriate steps before being implemented.  In general, any important change that 
theoretically increases risk to subjects constitutes an amendment.   
It is the responsibility of the Investigator to submit the amendment to the IRB for their 
approval; written approval should be obtained. The original signed copy of amendments will 
be kept in the Study File with the original Protocol.  It should be noted that where an 
amendment to the Protocol substantially alters the study design or the potential risks to the 
subjects, each subject’s consent to continue participation should be obtained. 
16.2 
Archiving of Records 
All source documents, CRFs and trial documentation will be kept by the Investigator for the 
appropriate retention period as stipulated by local regulations and ICH-GCP. 
16.3 
Study Report 
The Sponsor-Investigator or designee will produce the clinical study report at the completion 
of the study. In addition, it is intended that the outcome of the study will be submitted for 
publication in appropriate peer review journals and scientific meetings, as accepted. 
 
 
 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
17. 
REFERENCES 
1 Taylor CL, Carriquiry AL, Bailey RL, Sempos CT, Yetley EA. Appropriateness of the 
probability approach with a nutrient status biomarker to assess population inadequacy: 
a study using vitamin D. Am J Clin Nutr. 2013;97(1):72–78. 
doi:10.3945/ajcn.112.046094. 
 
2 Ginde AA, Liu MC, Camargo CA. Demographic Differences and Trends of Vitamin D 
Insufficiency in the US Population, 1988-2004. Arch Int Med. 2009;169(6):626–632. 
 
3 van Schoor NM, Lips P. Worldwide vitamin D status. Best Pract Res Clin Endocrinol 
Metab [Internet]. Elsevier Ltd; 2011;25(4):671–80. Available from: 
http://linkinghub.elsevier.com/retrieve/pii/S1521690X11000716 
 
4 Holick MF, Siris ES, Binkley N, Beard MK, Khan A, Katzer JT, et al. Prevalence of 
Vitamin D Inadequacy among Postmenopausal North American Women Receiving 
Osteoporosis Therapy. J Clin Endocrinol Metab [Internet]. 2005;90(6):3215–24. 
Available from: http://press.endocrine.org/doi/abs/10.1210/jc.2004-2364 
 
5 Glerup H, Mikkelsen K, Poulsen L, Hass E, Overbeck S, Thomsen J, et al. Commonly 
recommended daily intake of vitamin D is not sufficient if sunlight exposure is 
limited. J Intern Med. 2000;247(2):260–8.  
 
6 Tangpricha V, Pearce EN, Chen TC, Holick MF. Vitamin D insufficiency among free-
living healthy young adults. Am J Med. 2002;112(8):659–62. 
 
7 Business Group on Health. 2011. Injuries from Falls: Fact Sheet. 
www.businessgrouphealth.org/pub/f3115fe4-2354-d714-5195- 544ca84d399c (May 
2011). 
 
8 Hypponen E. Micronutrients and the risk of type 1 diabetes: vitamin D, vitamin E, and 
nicotinamide. Nutr Rev. 2004;62:340-347. 
 
9 Hypponen E, Laara E, Reunanen A, Jarvelin MR, Virtanen SM. Intake of vitamin D and 
risk of type 1 diabetes: a birth-cohort study. Lancet. 2001;358:1500-1503. 
 
10 Mithal A, Wahl DA, Bonjour J-P, Burckhardt P, Dawson-Hughes B, Eisman JA, et al. 
Global vitamin D status and determinants of hypovitaminosis D. Osteoporos Int 
[Internet]. 2009;20(11):1807–20. Available from: 
http://link.springer.com/10.1007/s00198-009-0954-6 
 
11 Kidney Disease: Improving Global Outcomes (KDIGO) CKD Work Group. KDIGO 2012 
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Disease. Kidney Int Suppl. 2013;3(1):1–150. 
 
12 Wang TJ, Zhang F, Richards JB, Kestenbaum B, van Meurs JB, Berry D, et al. Common 
genetic determinants of vitamin D insufficiency: a genome-wide association study. 
                                                 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
                                                                                                                                                       
Lancet [Internet]. Elsevier Ltd; 2010;376(9736):180–8. Available from: 
http://linkinghub.elsevier.com/retrieve/pii/S0140673610605880 
 
13 Holick MF, Binkley NC, Bischoff-Ferrari HA, et al. Evaluation, Treatment, and Prevention 
of Vitamin D Deficiency: an Endocrine Society Clinical Practice Guideline. J Clin 
Endocrinol Metab. 2011;96(7):1911–1930. doi:10.1210/jc.2011-0385. 
 
14 IOM (Institute of Medicine). Dietary Reference Intakes for Calcium and Vitamin D. (Ross 
AC, Taylor CL, Yaktine AL, Del Valle HB, eds.). Washington DC: The National 
Academies Press; 2011. doi:10.1542/peds.2012-2590. 
 
15 Hollis BW. Symposium : Vitamin D Insufficiency : A Significant Risk Factor in Chronic 
Diseases and Potential Disease-Specific Biomarkers of Vitamin D Sufficiency Vitamin 
D Intake : A Global Perspective of Current Status 1. J Nutr. 2005;135:317–322. 
16 Holick MF, Binkley NC, Bischoff-Ferrari HA, et al. Evaluation, Treatment, and Prevention 
of Vitamin D Deficiency: an Endocrine Society Clinical Practice Guideline. J Clin 
Endocrinol Metab. 2011;96(7):1911–1930. doi:10.1210/jc.2011-0385. 
17 Heaney RP, Recker RR, Grote J, Horst RL, Armas LAG. Vitamin D 3 is more potent than 
vitamin D 2in humans. J Clin Endocrinol Metab. 2011;96(3):E447–52. 
 
18 The Natural Standard Research Collaboration. Mayo Clinic Drugs and Supplments 
Vitamin D [Internet]. [cited 2015 Nov 17]. Available from: 
http://www.mayoclinic.org/drugs-supplements/vitamin-d/dosing/hrb-20060400 
 
19 Ish-Shalom S, Segal E, Salganik T, Raz B, Bromberg IL, Vieth R. Comparison of Daily, 
Weekly, and Monthly Vitamin D3 in Ethanol Dosing Protocols for Two Months in 
Elderly Hip Fracture Patients. J Clin Endocrinol Metab [Internet]. 2008;93(9):3430–5. 
Available from: http://press.endocrine.org/doi/abs/10.1210/jc.2008-0241 
 
20 Souberbielle J-C, Prié D, Courbebaisse M, et al. Actualité sur les effets de la vitamine D et 
l’évaluation du statut vitaminique D. Rev Francoph des Lab. 2009;2009(414):31–39. 
doi:10.1016/S1773-035X(09)70087-5. 
 
21 Rouillon V, Dubourg G, Gauvain J-B, et al. Vitamin D insufficiency: evaluation of an oral 
standardized supplementation using 100,000 IU vials of cholecalciferol, depending on 
initial serum level of 25OH vitamin D. Joint Bone Spine. 2012;79(4):399–402. 
doi:10.1016/j.jbspin.2011.09.004 
 
22 Holick MF. Vitamin D Deficiency. N Engl J Med. 2007;357:266–281. 
 
23 Maclaughlin JA, Clark MB, Holick SA, et al. Photosynthesis of Previtamin D3 in Human 
Skin and the Physiologic Consequences. Science (80- ). 1980;210(4466):203–205. 
 
24 Webb AR, de Costa B HM. Sunlight Regulates the Cutaneous Production of Vitamin D3 
by Causing Its Photodegradation. J Clin Endocrinol Metab 68882–887. 
1989;68(5):882–887. 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency 
                                                                                                                                                       
25 Holick MF, MacLaughlin J a, Doppelt SH. Regulation of cutaneous previtamin D3 
photosynthesis in man: skin pigment is not an essential regulator. Science. 
1981;211(4482):590–593. doi:10.1126/science.6256855. 
26 Kostoglou-Athanassiou I, Athanassiou P, Gkountouvas A, Kaldrymides P. Vitamin D and 
glycemic control in diabetes mellitus type 2. Ther Adv Endocrinol Metab. 
2013;4(4):122–8. doi:10.1177/2042018813501189. 
 
27 ICH Harmonized Tripartite Guideline: Guideline for Good Clinical Practice E6(R1) (10 
June 1996). Available from: 
http://www.ich.org/products/guidelines/efficacy/article/efficacy-guidelines.html 
 

University of Minnesota 
Protocol VITD-001 
 Topical Vitamin D in Vitamin D Deficiency