An Open-Label Study to Evaluate DPCP Ointment for the Treatment of Alopecia Areata
Phase: PHASE3
Conditions: Alopecia Areata
Sponsor: University of Minnesota

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Title:   
An Open-Label Study to Evaluate DPCP Ointment for the 
Treatment of Alopecia Areata 
 
 
Study Drug: 
Diphenylcyclopropenone Ointment 
 
Provided by: 
 
Hapten Pharmaceuticals LLC 
 
151 Merriweather Road 
 
Grosse Pointe Farms, MI 48236 
 
313-300-8454 
 
michaelburnsus@aol.com 
 
 
Sponsor:  
National Alopecia Areata Foundation 
 
65 Mitchell Boulevard, Suite 200-B 
 
San Rafael, CA 94903 
 
Phone: 415-472-3780 
 
Fax: 415-480-1800 
 
E-mail: info@naaf.org 
 
 
Principal Investigator: 
Maria Hordinsky, M.D. 
 
612-625-8625 
 
Hordi001@umn.edu 
 
Co-Investigator: 
Ronda Farah, M.D. 
 
 
 
 
Trial Site: 
Department of Dermatology 
 
University of Minnesota 
 
420 Delaware St. SE, MMC 98 
 
Minneapolis, MN 55455 
 
Version Date: 
Version 8 
 
February 29, 2024 
 
 
CONFIDENTIAL 
This is a University of Minnesota clinical research trial document that contains confidential 
information.  It is intended solely for the recipient clinical investigator(s) and must not be 
disclosed to any other party.  The material may be used only for evaluating or conducting clinical 
investigations; any other proposed use requires written consent from the University of 
Minnesota.  

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Table of Contents 
Study Summary 
       vii 
1. INTRODUCTION 
               1 
2. OBJECTIVES 
               5 
3. STUDY DESIGN 
               5 
Screening Visit 
           9 
Sensitization and Baseline Sample Collection Visit, Day -16 
           10 
Dose Determination Visit, Day -2 (10 to 14 days post-sensitization) 
            10 
Challenge Visit, Day 0 
11 
Post-First Treatment Sample Collection Visit and Second Treatment, Day 3 
11 
Treatment Visits, Twice per Week for Weeks 1-18 
12 
Investigator Assessment Visits (Weeks 4, 8, 12, and 16) 
12 
Regrowth Sample Collection Visit 
12 
Follow-up Visit, Week 18 
12 
Follow-up Visit, Week 20 
13 
Early Termination Visit 
13 
4. RATIONALE FOR STUDY DESIGN 
13 
4.1 Trial Treatment Descriptions 
14 
5. SELECTION OF STUDY POPULATION 
15 
5.1 Inclusion Criteria 
15 
5.2 Exclusion Criteria 
16 
6. SUBJECT DISPOSITION AND QUALITY ASSURANCE CRITERIA 
17 
6.1 Subject Withdrawal Criteria 
17 
6.2 Quality Assurance 
17 
7. TREATMENT OF SUBJECTS 
18 
7.1 Enrollment 
18 
7.2 Formulation 
18 
7.3 Dosage, Administration and Storage 
19 
7.4 Treatment Schedule 
19 
Pretreatment Period 
19 
Treatment Periods 
19 

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Follow-up Period 
19 
7.5 Concurrent Medications / Therapies 
19 
7.6 Prohibited Concomitant Medication and Treatment 
20 
8. EFFICACY ANALYSIS 
20 
8.1 Primary Efficacy Endpoints 
        20 
8.2 Exploratory Endpoints 
20 
9. ASSESSMENT OF SAFETY 
21 
9.1 Adverse Events (AE) 
21 
9.2 Baseline Medical/Preexisting Conditions 
22 
9.3 Procedures for Eliciting, Recording, and Reporting Adverse Events 
22 
9.4 Special Reporting Situations 
23 
9.5 Post Treatment Follow-Up 
23 
9.6 Post Adverse Event Follow-Up 
23 
9.7 Laboratory Tests 
23 
10. STATISTICAL METHODS 
24 
10.1 Data to be Analyzed 
24 
10.2 Determination of Sample Size 
24 
 Exploratory Endpoints 
25 
10.3 Blinding 
25 
10.4 Subject Disposition 
25 
10.5 Handling of Dropouts and Missing Data 
26 
11. DIRECT ACCESS TO SOURCE DATA/DOCUMENTS 
26 
11.1 Overview 
26 
11.2 Reporting and Recording of Data 
26 
12. ETHICAL CONSIDERATIONS 
26 
12.1 Informed Consent 
26 
12.2 Institutional Review 
27 
12.3 Ethical Considerations 
27 
15. FINANCING AND INSURANCE 
27 
16. REFERENCES 
27 
17. APPENDICES 
31 
APPENDIX A: Flowchart of Study Procedures 
32 
APPENDIX B: Instructions for Severity OF Alopecia Tool Assessments 
34 
Part I: The Severity of Alopecia Tool (SALT) 
35 
APPENDIX C: Adverse Event Report Log 
36 

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APPENDIX D: Serious Adverse Event Report 
37 
APPENDIX E: Demographics & Baseline Assessment 
40 
APPENDIX F: Treatment History Form 
41 
APPENDIX G: Physician Assessment of Alopecia 
42 
APPENDIX H: Subject Hair Loss Assessment 
43 
APPENDIX I: Sensitization Assessment 
44 
APPENDIX J: Study Completion Form 
46 
 
 
 

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List of Abbreviations 
AA 
Alopecia Areata 
AE 
Adverse Event 
BCC 
Basal Cell Carcinoma 
CFR 
Code of Federal Regulations 
CHS 
Contact Hypersensitivity 
CRF 
Case Report Form 
CTCAE 
Common Terminology Criteria for Adverse Events 
DNCB 
Dinitrochlorobenzene 
DPCP 
Diphencyprone, Diphenylcyclopropenone 
DTH 
Delayed Type Hypersensitivity 
FDA 
Food and Drug Administration 
HgB 
Hemoglobin 
HIPPA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
ICH 
International Conference on Harmonization 
IDS  
Investigational Drug Services Pharmacy 
IRB 
Institutional Review Board 
MCRU 
Masonic Clinical Research Unit 
MSD 
Meso Scale Discovery 
PAA 
Physician Assessment of Alopecia 
PBMCs 
Peripheral Blood Mononuclear Cells 

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PUVA 
Psoralen Plus Ultraviolet A 
SADBE 
Squaric Acid Dibutyl Ester 
SALT 
Severity of Alopecia Tool 
SCC 
Squamous Cell Carcinoma 
SHLA 
Subject Hair Loss Assessment 
SST 
Serum Separator Tube 
TSH 
Thyroid Stimulating Hormone 
UPIRTSO 
Unanticipated Problems Involving Risk to Subjects or Others 
USP-NF 
The United States Pharmacopeia and The National Formulary 
UVB 
Ultraviolet B 
 
 
 
 

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Study Summary 
 
Protocol Title 
An Open Label Study to Evaluate DPCP Ointment for the Treatment of 
Alopecia Areata 
Study Drug 
Diphenylcyclopropenone (DPCP) Ointment 
Indication 
Alopecia Areata 
Objectives 
Primary Objective: Determine the efficacy of DPCP ointment for the 
treatment of alopecia areata. 
Secondary Objectives:  
• Assess the safety of DPCP ointment in alopecia areata subjects 
• Determine the best starting dose range for future studies 
Exploratory Aims: To evaluate the association between cytokine and 
keratin profiles, Cytomegalovirus UL16 Binding Protein (ULBP) 
expression and hair regrowth. 
Study Design 
This is an open labeled study to determine the response and 
characteristics, safety and efficacy, of the proprietary DPCP ointment 
composition as a topical immunotherapeutic agent for the treatment of 
extensive alopecia areata.   
Up to 10 subjects with 76% to 99% scalp hair loss that, in the opinion of 
the PI, are eligible for treatment with DPCP will be enrolled. Patients 
will be recruited from the new and current patient population seen for 
alopecia areata through the University of Minnesota Medical Center 
Dermatology Clinic as well as through partnership with the National 
Alopecia Areata Foundation Clinical Trials Network, and various media 
outlets. 
 
The products to be evaluated are as follows:  
• 0.05 mL of 0.4% DPCP (sensitization dose) applied topically to 
the inner aspect of the upper right arm. (“Sensitization and 
Baseline Sample Collection Visit”) If sensitization is not attained 
after 10 to 14 days, the procedure will be repeated once. 
• 0.05 mL four dose concentrations of DPCP ointment (0.04, 0.08, 
0.12, 0.2%) prepared through dilutions of 0.4% DPCP Ointment 
with the 0.04% DPCP Ointment (applied to the inner aspect of the 
left upper thigh) and these will be applied sequentially starting 
with the lowest concentration and, if no sensitization is attained, 
sensitization to the next lowest dose will be attempted. This will 
be repeated until either the participant has a reaction and that 
concentration will be their dose for the duration of the study or 
until all concentrations have been unsuccessful in causing a 
reaction. If this is the case, the participant will be discontinued 

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from the study. (“Dose Determination Visit”; 10-14 days after the 
Sensitization Visit) 
 
The subject will return 72 hours after the application of each dose (0.04, 
0.08, 0.12, 0.2%) for an assessment by the PI or co-PI. As described 
above, the lowest dose will be applied first and assessed. If no reaction, 
the next lowest dose will be applied, etc, until adequate sensitization is 
attained. The weakest strength concentration that caused a minimal 
reaction will be used throughout the study. Once adequate sensitization 
has been obtained, 0.75-1 g of the treatment drug in that dose will be 
applied in a thin film that covers the entire scalp. 
The treatment drug will be applied by the PI, a trained study coordinator, 
or a staff member of the Clinical Research Unit at the University of 
Minnesota. Eligible subjects may begin receiving the study drug 
immediately after enrollment and screening.  
The estimated duration of the study is 22-24 weeks. See Appendix A for 
the schedule of visits. There will be a total of 42 scheduled visits, 
beginning with a screening visit followed by a single sensitizing dose of 
study drug at baseline (Day -16) and an initial dose determination 
application of 0.04% DPCP at Day -2. Additional visits could include 
another sensitization attempt as well as dose determination visits for the 
.04, 08, 0.12, and 0.2% DPCP.  
Subjects will undergo twice weekly (+/- 2 days) topical applications of 
an ointment formulation of DPCP during Weeks 1-18. Application will 
be done twice per week. The drug application will be performed by a 
board certified dermatologist for the first two treatment visits (Days 0 
and 3; Visits 4 and 5), after which a trained member of the Clinical 
Research Unit staff or a trained study coordinator will apply the drug 
twice each week (Weeks 2-18; Visits 6-39). The investigator will attend 
one treatment visit in the Clinical Research Unit each month to evaluate 
the extent of hair loss and hair growth (Weeks 4, 8, 12, and 16).  
Scalp biopsy specimen collection will be performed at baseline (Day -16, 
Visit 2), 72-96 hours after the first treatment (i.e., challenge) application 
(Day 2, Visit 5), 72-96 hours after the final application (Week 18 +3d, 
Visit 39) or during treatment when the study subject is determined to 
have achieved >=50% hair regrowth. 
Three scalp biopsy samples will be collected by a board certified 
dermatologist. When possible, each sample will be taken from a balding 
area near an area with hair preferably in a non-androgen dependent site 
of the scalp. One of the three samples will be frozen in OCT for 
histologic examination and immunohistochemical studies. A second 
biopsy sample from an adjacent area will be placed in 10% buffered 

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formalin for histologic examination and assessment of inflammation and 
hair follicle differentiation. The third sample from each area will 
immediately be placed in RNAlater (Qiagen, Valencia, CA) for cytokine 
expression analyses. 
Peripheral blood collection for the study will include one tube of blood in 
a serum separator tube (red top SST tube) and one tube of blood in a 
PAXgene tube for RNA isolation. All blood samples will be transported 
to the Dermatology laboratory at the University of Minnesota for 
immediate processing and storage for additional biomarker studies. 
 
Research coordinators will be involved in patient recruitment, contact, 
and scheduling. The PI and research coordinators will all be involved in 
the collection, analysis, and reporting of collected data. 
Concomitant 
Medications 
Medications with no proven effect on the immune system will be 
acceptable. 
Study Population 
This study includes subjects ages 18 and above. Patients of the 
University of Minnesota Dermatology Clinic who meet the 
inclusion/exclusion criteria will be given the opportunity to participate in 
this study, as will select participants in the National Alopecia Areata 
Foundation Clinical Trials Network. Dr. Hordinsky has several Hair 
Disease clinics and appropriate patients will be offered the opportunity to 
participate in this study. 
Sample Size 
Up to 10 subjects 
Study Sites 
University of Minnesota 
Duration of Subject 
Participation 
4-5 month treatment period consisting of a sensitization visit, dose 
determination visit (at least one, up to 4 total), 36 treatment applications 
at twice weekly (+/- 2 days) intervals, a  post-treatment safety evaluation 
and biopsy collection procedure 72-96 hours after the last treatment visit, 
and a visit 7-14 days after the final biopsy collection for suture removal. 
Outcome Measures 
Primary Efficacy Endpoints 
In this study, the Severity of Alopecia Tool (SALT) will be used. The 
SALT score is a global AA severity score based on scalp hair loss. To 
determine the SALT score, 4 main areas are assessed with these 
representing 18%, 18%, 40%, and 24% of the total scalp surface area. 
50% scalp hair regrowth based on SALT score can be expressed as 
SALT50.  
The primary endpoints ranked below in order of importance are as 
follows: 

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1. The proportion of subjects that has ≥ 75% decrease in their SALT 
score (SALT75), as compared to baseline at the completion of the 
study or at any time during the study. 
2. The proportion of subjects that has ≥ 50% decrease in their SALT 
score (SALT50), as compared to baseline at the completion of the 
study or at any time during the study.   
Exploratory Endpoints 
1. To identify peripheral blood and scalp skin biomarker profiles 
that characterize a response to DPCP treatment, enabling the 
future potential for predicting individual responses to treatment 
and the likelihood of a sustained response. 
2. Subject assessment of their hair loss using the subject hair loss 
assessment scale.  
Estimated Start 
03/15/2023 
Estimated Finish 
12/31/2024 
Safety 
All regulations stated in 21 CFR Parts 50, 56, and 312 and outlined in the 
ICH Guidelines for Good Clinical Practice will be adhered to throughout 
this trial. Concomitant medications and adverse events will be monitored 
and tracked.  
Adverse events (AEs) may be observed or elicited by the Investigator 
and/or volunteered by the subjects. AEs will be graded on a four-point 
scale (mild, moderate, severe, or life-threatening). AEs will be recorded 
on the Adverse Event Report (Appendix C) and documented in the 
subjects’ files. Serious adverse events (SAEs) will be documented on the 
Adverse Event Report (Appendix C) and the Serious Adverse Event 
Report Form (Appendix D). SAEs will be reported to study drug 
manufacturer, University of Minnesota IRB, and the FDA. 
Statistical 
Considerations 
Statistical Considerations: The proportion of subjects with ≥75% and 
≥50% decrease in the SALT score (at any time during treatment; 
endpoints 1 and 2) will be described using the fraction achieving the 
desired decrease and the associated confidence interval, calculated by the 
Agresti-Coull exact method. To describe change in SALT score, we will 
use the average change and associated confidence interval.   
Statistical power: For a total sample size of 10 we anticipate 6 or 8 
participants providing data. For changes in binary outcomes, for 6 
participants, the Agresti-Coull 95% confidence intervals for the 
proportion having an effect range in width from 0.44 (0 to 0.44 if 0 of 6 
have an effect or 0.56 to 1 if 6 of 6 have an effect) to 0.62 (i.e., 0.19 to 
0.81 if 3 of 6 have an effect).  For 8 participants, the Agresti-Coull 95% 
confidence intervals for the proportion having an effect range in width 
from 0.37 to 0.56.   

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Exploratory Endpoints 
To identify scalp skin biomarker profiles that characterize a response to 
DPCP treatment, enabling future potential for predicting individual 
responses to treatment and the likelihood of a sustained response. The 
primary outcomes for this analysis are levels of T-cell and DC markers, 
which will be analyzed using paired t-tests. To estimate power, 
preliminary data were obtained from a pilot study (Fuentes-Duculan et al. 
Abstract, ISDS, November, 2014). Assuming the standard deviations are 
as reported in that abstract, for comparing pre- vs. post-treatment, for 6 
participants we have 80% power to detect a 14-fold change (using two-
tailed alpha = 0.05);  for 8 participants the detectable difference is 8-fold. 
For the analysis of various biopsy parameters, Dr. Krueger will be 
supported by a statistician in his group. 
For the change in subject VAS score, the confidence interval width will 
be 2.1 standard deviations for 6 participants and 1.7 standard deviations 
for 8 participants; we lack information needed to supply a standard 
deviation describing variation between persons in this change.   

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1. INTRODUCTION 
Alopecia Areata (AA): AA is an autoimmune disease characterized by immune targeting 
of anagen hair follicles (growing stage of the hair cycle) and subsequent disruption of the hair 
cycle.1 The first National Health and Nutrition Examination Survey conducted from 1971 
through 1974 found the prevalence of AA in the United States to be 158 per 100,000 persons, 
which is roughly 0.1 to 0.2% of the population.2 AA presents as patchy AA or extensive, 
involving all of the scalp (alopecia totalis) or scalp and body hair (alopecia universalis). It is 
estimated that AA accounts for approximately 4% of visits to dermatologists. AA has a 
considerable impact on quality of life and can be a devastating disease.3–7 Support for a 
multifactorial inheritance pattern comes from many sources.8–11 Transplantation studies in 
severe-combined immunodeficient mice show T-cell immunity is directly involved in the 
disease.12 A hallmark of active AA is the presence of peribulbar lymphocytes around anagen 
follicles and the presence of killer CD8 T cells attracted by NKG2DL.13 Immune privilege may 
be lost in AA.14,15 A genome-wide association study showed the involvement of both innate and 
acquired immunity.11 More recently investigators have also demonstrated in scalp biopsies from 
AA patients with mild to extensive disease that lesional AA samples had an “inflammatory” AA 
subset characterized by higher expression of T-cell (CD4, CD2, CD3D, CD3G, and CD28) and 
dendritic cell markers (CD11c, CE1b, CD83 and CD86). More extensive scalp involvement was 
also found to be associated with higher genomic dysregulation and multiple lines of evidence 
suggest that there are shared genetic risk factors between AA and other autoimmune diseases.11 
Treatments: Many therapies are available off-label and current treatment choices are 
frequently based on disease extent, duration, and age of the patient.16–19 In June 2022, the FDA 
approved an oral janus kinase inhibitor for the treatment of AA, however, this medication is 
associated with immunosuppression and potential risk. (Ref 20-21) Assessing efficacy of 
treatments for AA is complicated because there are few published randomized controlled trials. 
However, there are several uncontrolled trials and reports with non-ideal criteria to evaluate 
treatment progression and long-term follow-up is not included in most of the published works. In 
1999 and 2004 the AA Investigational Assessment Guidelines were published and have been 
subsequently applied in up to 30% of new published investigations, providing some 
standardization to the description of disease extent and percent improvement in response to 
therapy.20,21 
 
Sensitizers  
Three different topical sensitizers have been used to treat alopecia areata with some 
evidence of efficacy. These three include dinitrochlorobenzene (DNCB), diphencyprone (DPCP), 
and squaric acid dibutyl ester (SADBE). The theory of immunomodulation was proposed by 
Happle et al in 1986 when they showed that skin treated with topical sensitizers showed changes 
in the peribulbar CD4/CD8 lymphocyte ratio.22  In 2006 Herbst et al reinforced and 
complemented the theory providing evidence that long term treatment with a contact sensitizer 
allows for the recovery of the hair follicle by driving autoreactive T cells into activation-induced 
cell death.23 
DNCB was first used for alopecia areata in 1976,24 but due to concerns over the 
mutagenic properties of the drug in Salmonella enteritides serotype Typhimurium and its 
absorption through the skin with ultimate excretion in urine, its use has in large part been 
discontinued.25 Because SADBE is stated to be relatively unstable in acetone solution , DPCP is 

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the most commonly used sensitizer in clinical practice.26,27 Happle et al enthusiastically 
introduced the use of DPCP for the treatment of alopecia areata when they reported that 67% of 
treated patients had a satisfactory response to this treatment.26 
However, in a larger study of 139 patients completed by the same investigator, a response 
rate of 50.4% was subsequently reported.28 Some years later a review of 17 reported cases series 
concluded that though 50-60% of patients may achieve a response to DPCP, the range of 
reported responses was very wide, between 9 to 87%.29  In a follow-up study, Gordon et al 
included a total of 32 patients and demonstrated that only 9 patients maintained regrowth without 
further DPCP treatment for an average of 19.8 months, while 9 had poor regrowth with 
continued use of the drug and 5 discontinued the treatment.30  
A review of AA treatments in 2003 presented the evaluation of the 17 most significant 
controlled trials from 1983 to 2001, using DPCP and SABDE studies to test the efficacy of 
treating AA.31 The response rate of treatments represented by cosmetically acceptable hair 
regrowth varied from 29% to 78%. The authors suggested that these differences could be 
explained by the different extent and durations of alopecia areata present in the patient 
populations prior to the treatment in each study and by the different treatment methodologies.31 
In yet another retrospective chart review of patients from 1996 to 1999, 33% were found to attain 
cosmetically acceptable or total regrowth with topically applied SADBE.32 
Additional studies have been performed more recently around the globe, and have 
continued to support the efficacy of contact sensitizers for some patients with AA. An Indian 
study in 2006 involved 70 patients treated with SADBE for 4 months. The overall success rate 
was 43%. For subjects with < 50% of scalp involvement the rate was 68%, however for those 
with > 50% involvement the success rate was 29%. Additionally 81% of the responders (21 in 
26) had relapse occurrence.33 In a study completed in Greece 64 patients with alopecia areata 
were sensitized with 2% DPCP and treated with graduated concentrations adjusted to maintain 
erythema and pruritus on the scalp for 48 hours. The follow up was 2 years. A total of 83.3% of 
the subjects responded to therapy, although a significant difference in responses was noted and 
varied between regrowth of vellus hair, sparse pigmented terminal hairs, terminal hairs in 
patches of alopecia as well as regrowth of terminal hair on the entire scalp. Relapses were noted 
to occur in approximately 70% of the patients.34 A Chinese study published in 2012 investigated 
the efficacy of DPCP in treating 31 steroid non-responders with extensive AA (defined as >30 
percent hair loss) after 6 months of weekly treatment. Similar to the Greek study, participants 
were sensitized with 2% DPCP and treated with graduated concentrations adjusted until an 
erythematic reaction was maintained for 48 hours. The authors reported 37.9% of participants 
exhibited complete (n = 4) or partial (n = 7) remission of AA, with a 69.2% relapse rate in the 
subsequent 18 months (n = 9/13).35 
The side effects of topical immunotherapy include the desired mild dermatitis inherent to 
the treatment and in 2-5% of patients vesicular and bullous reactions at the initiation of treatment 
before appropriate individual concentrations are determined, dissemination of allergic contact 
dermatitis, urticaria or erythema multiforme-like reactions and pigmentary disturbances such as 
postinflamatory hyperpigmentation. Notably, no long term side effects have been reported after 
18 years use of DCPC and 21 years of SADBE treatments worldwide.36  
 
Cytokine Profiling 
Studies designed at investigating the role of cytokine expression during active disease 
have resulted in an increased comprehension of the molecular basis of disease, as well as, the 

Page 3 
 
potential of more targeted therapies. A study by Czarnowicki et al. explored the extent of T-cell 
activation in atopic dermatitis (AD) and psoriasis and found that patients with AD had higher 
and more persistent activation of central and effector memory T cells which may explain the 
systemic and frequent disease in these patients. This type of study allows for identification of 
cytokine profiles that characterize a disease.37 
A previous clinical study that investigated the role of cytokines, specifically IL-17A, 
found that this cytokine plays a role in the pathogenesis of psoriasis. Krueger et al. found that 
Th17 cell cytokine interleukin-17 (IL-17) is likely causing inflammation and epidermal 
hyperplasia in psoriasis. This study combined an experimental therapeutic drug trial with 
cytokine profiling. Skin biopsies were obtained from participants undergoing subcutaneous 
injections of ixekizumab. A combination of histology, RT-PCR, and a gene array were used to 
analyze cytokine concentrations in order to better understand their role and contribution to 
disease. IL-17 was identified as the major “driver” cytokine for the disease phenotype in 
psoriasis. This study provides support for the investigation of the role of cytokines as an 
indicator and biomarker of disease in psoriasis and other related diseases including alopecia 
areata.38 
Recent studies have found differences in the transcriptomes between lesional and non-
lesional areas of disease. Specifically, a greater immune and keratin dysregulation corresponding 
to greater scalp involvement (>25%) was found. Activation of cytokines, TH2, TH1, IL-23, and 
IL-9/TH9 were identified to be a signature for alopecia areata.39 A study also performed at the 
Krueger Laboratory identified dysregulation of T cell expansion in a subtype of psoriasis specific 
to the Asian population. IL-17A and IL-17- were found to be highly expressed specifically in 
Asian small plaque psoriasis, compared to Western large plaque psoriasis. Similar differences 
worldwide in alopecia areata remain to be reported. In summary, cytokine profiling studies 
allows for a deeper understanding of the main mechanism of disease development.40  
 
DPCP Non-clinical Studies 
Although it has not received FDA or EMA approval to date, several investigator 
sponsored trials and research programs have generated data to support the safe use of DPCP for 
the treatment of several indications, including warts, alopecia areata and cutaneous metastases of 
melanoma. 
Toxicity and Safety Studies 
Genetic Toxicity 
 
 
Two standard in vitro genetic toxicity studies, a bacterial mutagenicity assay (Ames test) 
and a chromosomal aberration assay, were conducted with DPCP obtained from the API 
supplier, Carbosynth.  Both tests were negative under the conditions of the assays, attesting to 
the absence of a risk for genetic toxicity.  In addition, an in vivo micronucleus assay was 
performed to evaluate DPCP for in vivo clastogenic activity and/or disruption of the mitotic 
apparatus by detecting micronuclei in polychromatic erythrocyte cells (mnPCEs) in mouse bone 
marrow.  DPCP administered by intraperitoneal injection did not induce a significant increase in 
the incidence of mnPCEs in bone marrow at any dose tested.  DPCP was concluded to be 
negative in an in vivo micronucleus test designed to test for in vivo clastogenic activity. 
DPCP Ointment has been evaluated in studies carried out by Hapten Pharmaceuticals 
under BLA 10,223 and IND 77,043. DPCP Ointment is also being evaluated in the treatment of 

Page 4 
 
cutaneous metastases of melanoma in Investigator Sponsored individual patient INDs.  The 
status and results to date of these studies are summarized below. 
Diagnostic Use as a Measure of Immune Competence 
A clinical study was conducted by Pharmaderm Associates, LLC (now Hapten 
Pharmaceuticals) to determine if DPCP can be used as a diagnostic for assessing 
immunocompetency in HIV-positive subjects.41 Forty HIV-positive subjects were enrolled and 
divided into four groups to be treated with one of the following DPCP concentrations (0.4, 0.2, 
0.1, or 0.05%). The DPCP was formulated in 50/50 polysorbate 80/isopropyl myristate and 
applied on a one square inch latex-free adhesive pad to the medial surface of the right arm and 
left in place for 48 hours.  Skin reactivity was assessed weekly on a scale of 1 to 4 with ≥ 2 
considered positive for immunocompetence.  The skin reactivity scores correlated with CD4 T 
cell counts, suggesting DPCP could be used as a simple test for immunocompetency. This study 
also clearly demonstrated that 0.4% DPCP can successfully elicit a DTH response in most 
patients with CD4 T cell counts in the normal range.  In these subjects there were no skin 
reactivity scores of greater than 2 (erythema and induration) even at the highest dose of DPCP of 
0.4%. 
Healthy Volunteer Biomarker Study 
In a study by Gulati et al., 11 healthy volunteers were sensitized with sensitizing DPCP 
ointment (0.4%) followed by challenge applications of treatment DPCP ointment (0.04%) as well 
as topical placebo applications. The treated sites were biopsied at different time points for 
comprehensive cellular and molecular profiling.  All volunteers tolerated treatment well, with the 
only adverse events being the expected inflammatory skin reactions to DPCP ointment. No 
serious adverse events, unexpected adverse events, or other types of adverse events occurred.42 
The analysis of biopsy samples included gene array approaches, and ~7,500 mRNA 
transcripts were modulated during the peak inflammatory responses induced by DPCP.  These 
responses were characterized by high expression of cytokines that define all major effector T cell 
subsets. The study suggests the presence of active regulatory mechanisms in the resolution of 
inflammation induced by DPCP, proving the utility of using DPCP to study mechanisms of 
immune regulation in human skin. 
Cutaneous Metastasis of Melanoma 
There have been two investigator initiated individual patient INDs in which DPCP 
ointment was evaluated for treatment of cutaneous metastases of melanoma. Gulati et al. (2016a) 
demonstrated dramatic reduction of cutaneous metastases in a patient who received three months 
of combined pembrolizumab (an immune checkpoint inhibitor) and DPCP therapy (alternating 
0.4% and 0.04% concentrations applied twice weekly). Treatment using either agent alone 
produced mixed results. The patient also developed vitiligo on the contralateral leg not treated 
with DPCP, which suggests a synergistic immune response against melanocyte lineage cells.43 
 
Gulati (2016b) characterized the immune response to DPCP in a cohort of six patients 
with cutaneous melanoma metastases. Immune reactions induced by DPCP were characterized 
using both immunohistochemical (IHC) and gene expression approaches. The authors reported 
that five patients achieved partial or complete regression of their cutaneous metastases after 
minimum 14 weeks (range: 14-28 weeks) of 0.4% DPCP treatment, applied twice weekly, as 
indicated by reduced IHC staining of the melanocyte marker MLANA. Further evidence of 

Page 5 
 
melanoma regression was demonstrated through gene expression microarray analysis. This 
analysis showed downregulation of hallmark melanoma and melanocyte gene expression post-
DPCP treatment compared to pre-DPCP treatment. The genes most significantly downregulated 
included five “melanoma signature” genes PRAME, TYR, OCA2, DCT, and MLANA, which were 
downregulated 12- to 22-fold. Notably, one patient left the trial before sensitization could be 
induced, and was not included in these results.44  
Common Warts 
A study was carried out to evaluate DPCP ointment for the treatment of common warts 
(verruca vulgaris).  This Phase 2a trial, sponsored by PharmaDerm Associates, LLC (later 
Hapten Pharmaceuticals), was a randomized, double blind, placebo controlled study to determine 
the safety and efficacy of DPCP ointment as a topical immunotherapy for the treatment of 
common warts.  The study enrolled adults with at least one 5-20 mm non-genital wart.  The 
treatment consisted of a sensitization dose applied to the inner arm and one target wart with 
Sensitizing DPCP ointment (0.4%) or placebo (ointment with no DPCP) on Day 0, followed by 
seven (7) weekly administrations of treatment DPCP ointment (0.04%) or placebo applied to 
each target wart (up to 4 per subject) beginning on Day 15.  A final follow-up examination was 
carried out two weeks after the last treatment dose. The response to sensitization was graded at 
the Day 15 visit on a scale of 0-4.  Adverse reactions, immunotherapeutic response and efficacy 
using IGAS and Lesion Surface Area scores were assessed during each visit throughout the 
treatment and follow-up period. 
The percent clearance by wart was calculated for the ‘Per Protocol’ population 
(completing all treatments and assessment visits).  At least 50% clearance was reported in 47.6% 
of the DPCP ointment treated wart lesions compared to 6.7% of the wart lesions treated with 
placebo ointment. This result is statistically significant (p = 0.011) indicating a clinical benefit in 
the clearance of warts with DPCP ointment. 
Drug-related adverse events reported in the study were local reactions due to the 
sensitization and challenge responses in the skin. The most commonly reported adverse events 
(AEs) were rash, pruritus, and erythema.  The highest rate of reported AEs occurred after the 
sensitization step (reported at the Day 15 visit). 
2. OBJECTIVES 
Primary Objective: To determine the efficacy of DPCP ointment for the treatment of alopecia 
areata. 
 
Secondary Objectives: 1) To assess the safety of DPCP ointment in alopecia areata subjects. 
2) To determine the best starting dose range for future studies 
 
Exploratory Aims: 1) To evaluate the association between cytokine and keratin profiles, 
Cytomegalovirus UL16 Binding Protein (ULBP) expression and hair regrowth. 
 
3. STUDY DESIGN 
• The products to be evaluated are 0.4% DPCP and four strengths of DPCP ointment (0.04, 
0.08, 0.12, 0.2%) (Add preparation)  Four dose concentrations of DPCP ointment (0.04, 
0.08, 0.12, 0.2%) will be prepared through dilutions of 0.4% DPCP Ointment (applied to 

Page 6 
 
the inner aspect of the left upper thigh) and these will be applied sequentially starting 
with the lowest concentration and, if no sensitization is attained, sensitization to the next 
lowest dose will be attempted. This will be repeated until either the participant has a 
reaction and that concentration will be their dose for the duration of the study or until all 
concentrations have been unsuccessful in causing a reaction. If this is the case, the 
participant will be discontinued from the study. (“Dose Determination Visit”; 10-14 days 
after the Sensitization Visit) 
  
  For sensitization, 0.05 mL of the 0.4% concentration will be applied topically to the right upper 
inner arm. If sensitization is not attained after 10 to 14 days, the procedure will be repeated once. 
After sensitization is established, the lowest treatment dose will be applied to the inner aspect of 
the upper left thigh. If sufficient sensitization is not attained, the next lowest treatment dose will 
be applied, etc, until sensitization is attained. If no sensitization is attained to any of the 4 
concentrations, the participant will be discontinued from the study. The treatment dose will be 
selected based on lowest concentration of DPCP that causes minimal reaction. This treatment 
dose will be used throughout the study. The treatment phase of the study will consist of twice-
weekly applications of 0.75-1 g DPCP ointment to the entirety of the scalp. Every four weeks 
during treatment, the investigator will assess hair loss as described below: 
 
1) Efficacy will be assessed as follows: 
a. Severity of Alopecia Tool (SALT) Score: Percentage of hair loss on the scalp will 
be determined using the SALT score, a global alopecia areata severity score based 
on scalp hair loss.20,21 The SALT score is calculated by visually examining four 
discrete areas of the scalp, measuring the percentage of terminal hair loss in each 
area, and then totaling the results (See Appendix B). 
b. Scalp Photography. 
c. Hair Pull Tests (positive or negative). 
 
2) Safety will be assessed by monitoring side effects at each visit and recording adverse 
events for each subject on the Adverse Event Report (Appendix C). Adverse events will 
be tracked for each subject from the time of the first sensitization does application of the 
drug until the subject completes the study. Adverse events will be assessed for severity 
by the investigator and graded on a four-point scale (mild, moderate, severe, or life-
threatening). Serious adverse events will also be recorded on the Severe Adverse Event 
Report (Appendix D), and the study drug manufacturer and the U of MN IRB, and the 
FDA will be notified. Skin reaction scores will be recorded during the sensitization 
assessment (Appendix I) to monitor reactions, and may constitute an adverse event based 
on severity of reaction.  
 
3) Molecular and immunological characterization of the Alopecia Areata disease process 
and how it changes throughout the course of treatment with DPCP will be achieved 
through mRNA expression profiling in blood and scalp biopsy specimens, and 
histopathology analyses. 
 
4) Molecular predictors of positive and negative response to DPCP treatment will be 
identified through expression profiling targeted to mRNA transcripts for cytokines, T cell 

Page 7 
 
subsets, and hair keratins in scalp biopsy specimens. Peripheral blood mononuclear cells 
will be frozen and also serum frozen for future studies looking downstream for possible 
changes with DPCP in T-cell activation markers or circulating cytokines such as gamma-
interferon, IL-13, IL-17, and/or an increase in Tregs (CD25+/CD127- population or 
Foxp3+ population). This will not need to happen in blood if all the changes however are 
found to be happening in skin/scalp. 
 
3.1 Study Design  
This is an open labeled study to determine the response and characteristics, safety and efficacy, of 
the proprietary DPCP ointment composition as a topical immunotherapeutic agent for the 
treatment of extensive Alopecia Areata. The products to be evaluated are 0.4% DPCP and four 
strengths of DPCP ointment (0.04, 0.08, 0.12, 0.2%), with each dose determined for every 
participant individually. 
  
Up to 10 (minimum of 6) subjects with diagnosed extensive alopecia areata (76%-99% scalp hair 
loss as determined by the SALT score; see Appendix B, Part I) that are eligible for DPCP 
treatments will be consented and enrolled in the study. Patients with alopecia areata will be 
recruited from the new and current patient population seen in the University of Minnesota 
Medical Center Dermatology Clinics as well as through partnership with the National Alopecia 
Areata Foundation Clinical Trials Network. Potential subjects will also be recruited using flyers 
posted at the University of Minnesota and in surrounding communities. The study may be 
advertised using various media outlets, including newsletters, campus wide mailings, campus 
email announcements, the Dermatology department website, and listservs in the Twin Cities area 
(including the listserv for the Minnesota Alopecia Areata Support Group). All materials used in 
recruitment will be approved by the Institutional Review Board (IRB) prior to implementation. 
 
There will be a total of 42 scheduled visits, beginning with a screening visit. Subjects will 
undergo 38 scheduled topical applications of a an ointment formulation of DPCP, including a 
sensitizing dose of study drug at Day -16, an initial dose determining application at Day -2, 
followed by a challenge at Day 0, and twice weekly (+/- 2 days) treatment doses from Weeks 1-
18. Additional applications of DPCP could include another sensitization visit or additional dose 
determination visits for the higher concentrations, if necessary. There will be a scalp biopsy 
collection at baseline (Visit 2, Day -16), and visits for scalp biopsy collection 72-96 hours after 
the first treatment, and 72-96 hours after >50% hair regrowth or 72-96 hours after application of 
the final treatment dose. Suture removal will take place during treatment visits or scheduled 7-14 
days after each biopsy visit. Several questionnaires and forms will be completed throughout the 
duration of the study. 
 
Research coordinators will be involved in patient recruitment, contact, and scheduling. The PI and 
research coordinators will all be involved in the collection, analysis, and reporting of collected 
data. 
 
All visits will take place either in the M Health Clinical Research Unit (CRU), part of the 
Clinical and Translational Science Award at the University of Minnesota, or in the LCRU. There 
are no additional collaborating sites for this study.  
 

Page 8 
 
Schedule of Visits 
 
Visit 
Procedure 
1a 2b 3c 4d 5 
6-40e 
41 42f 
Informed consent 
X  
 
 
 
 
 
 
Screening for eligibility 
X  
 
 
 
 
 
 
Blood tests 
X  
 
 
 
X 
(1 time at week 
12) 
X 
 
Pregnancy test 
X  
 
 
 
X 
(1 time a month) 
X 
 
Vital signs 
X X  
 
X X (1 time a mo.) 
X 
 
Adverse events 
 
X X X X X 
 
X 
Assessment of hair loss 
X  
X  
 
X 
(1 time a month) 
X 
 
Photographs 
 
X X  
 
X  
(1 time a month) 
X 
 
Scalp biopsy procedure 
 
X  
 
X X  
(if 
50% 
regrowth) 
X 
 
Suture removal 
 
 
 
X  
X  
(as needed) 
 
X 
Blood draw 
 
X  
 
X X  
(if 
50% 
regrowth) 
X 
 
DPCP ointment - sensitizing 
 
X  
 
 
 
 
 
DPCP 
ointment 
– 
dose 
determination 
for 
each 
concentration: 0.04, 0.08, 0.12, 
0.20  
 
 
X  
 
 
 
 
DPCP ointment - challenge 
 
 
 
X  
 
 
 
DPCP ointment - treatment 
 
 
 
X X X  
(twice per week 
For 17 weeks, 
visit 40 will occur 
during week 18) 
 
 
Study completion 
 
 
 
 
 
 
 
X 
a. Screening visit 
b. Sensitization and baseline sample collection – visit can be repeated if participant not a responder to DPCP the first time. 
Blood draw and scalp biopsy procedure will only take place if sensitization has occurred and the participant is a responder 
to DPCP.  
c. Dose determination visit – see following schedule of events for more details.  
d. Challenge visit 
e. Participants will return twice per week for application of the previously selected dose of DPCP. Every four weeks during 
treatment, the investigator will attend the treatment visit to perform assessments of hair loss. 
f. 
Suture removal and study completion 
 
Dose Determination visit schedule:  

Page 9 
 
 
 
Visit 
Procedure 
31 32 33 34 
Adverse events 
X X X X 
Assessment of hair loss 
X  
 
 
Photographs 
X X X X 
DPCP ointment – 0.04% 
X  
 
 
DPCP ointment – 0.08% 
 
X  
 
DPCP ointment – 0.12% 
 
 
X  
DPCP ointment – 0.2% 
 
 
 
X 
 
1: DPCP ointment 0.04% concentration is the starting concentration for all participants  
2: DPCP ointment 0.08% will only be applied if participant does not have a strong enough response [defined as skin 
reaction score 1+] to the 0.04% concentration of DPCP. Thus, this visit will begin with an assessment of the sensitization 
reaction to the 0.04% dose, If there is not a strong enough response, 0.08% concentration DPCP will be applied to a 
different area of the inner upper left thigh in the same manner as the 0.005% dose. If the participant is a responder to the 
0.04% DPCP, patient will proceed to the challenge visit (visit 4) as described above.  
3: DPCP ointment 0.12% will only be applied if participant does not have a strong enough response [defined as skin 
reaction score 1+] to the 0.08% concentration of DPCP. Thus, this visit will begin with an assessment of the sensitization 
reaction to the 0.08% dose, If there is not a strong enough response, 0.12% concentration DPCP will be applied to a 
different area of the inner upper left thigh in the same manner as the 0.08% dose. If the participant is a responder to the 
0.08% DPCP, patient will proceed to the challenge visit (visit 4) as described above.  
4: DPCP ointment 0.2% will only be applied if participant does not have a strong enough response [defined as skin 
reaction score 1+] to the 0.12% concentration of DPCP. Thus, this visit will begin with an assessment of the sensitization 
reaction to the 0.12% dose, If there is not a strong enough response, 0.2% concentration DPCP will be applied to a 
different area of the inner upper left thigh in the same manner as the 0.12% dose. If the participant is a responder to the 
0.2% DPCP, patient will proceed to the challenge visit (visit 4) as described above.  
 
 
Screening Visit 
At the screening visit the PI will obtain signed, informed consent and HIPAA approval. 
Demographic information, weight and vital signs, medical history including concomitant 
medications will be recorded on the Demographics and Baseline Assessment form (Appendix 
D). Research personnel will review eligibility with the subject. Lifetime treatment history will 
also be recorded (Appendix E). A dermatologic exam will be performed to confirm that the 
extent of alopecia areata is 76-99% hair loss according to the SALT score (Appendix B, Part I). 
Laboratory tests will be drawn to measure TSH, Hemoglobin (HgB), and Vitamin D level, and 
thus to reduce the likelihood a common underlying medical problem that impacts hair growth 
exists prior to study participation. Additional laboratory tests will be draw as part of safety 
monitoring as described in section 9.7. Pregnancy test will be administered to all women of 
childbearing potential, and must be negative to participate. Safety laboratory results will be 
documented in the subjects’ medical record. 
 
An appointment for the sensitization visit will be scheduled with research personnel. If a 
washout period is required prior to treatment, the subject will return for the sensitization visit at 
the appropriate time. Otherwise, the research subject may immediately proceed to the 
sensitization part of the study or may be scheduled at a determined date in the future for the 
sensitization and baseline sample collection visit described below. 
 

Page 10 
 
Sensitization and Baseline Sample Collection Visit, Day -16 
Inclusion and exclusion criteria will be reviewed again, especially if there was a washout period 
between the screening and sensitization visits. Interim medical history, concomitant medications, 
and vital signs will be recorded on a Case Report Form (CRF). Assessments of hair loss, 
including the SALT score, the Physician Assessment of Alopecia (PAA; Appendix F), and the 
Subject Hair Loss Assessment (SHLA; Appendix G) will be completed by the physician or 
subject. Pregnancy test will be administered to all women of childbearing potential, and must be 
negative to participate. AEs (Appendix C) will be monitored and recorded. 
 
Three 4-mm scalp biopsy samples will be collected from perilesional scalp by a board certified 
dermatologist knowing that in some patients this site may be by just a few tufts of fibers. One of 
the three samples will be frozen in OCT for histologic examination and immunohistochemical 
studies. Those samples will be stored in a -80 degree freezer and batch shipped to Dr. Krueger 
and the Laboratory of Investigative Dermatology at Rockefeller University in New York, NY on 
dry ice every one to two months. A second biopsy sample from each area will be bisected and 
placed in 10% buffered formalin and Zamboni’s fixative for histologic examination and 
assessment of inflammation and hair follicle differentiation in the Dermatology Laboratory at the 
University of Minnesota. The third sample from each area will immediately be placed in 
RNAlater (Qiagen, Valencia, CA) and shipped to the Krueger Laboratory in the Laboratory of 
Investigative Dermatology at Rockefeller University in New York, NY for mRNA-level cytokine 
analyses. 
 
Peripheral blood samples will be collected by a trained staff member and will include 1 serum 
separator tube (SST) and 1 PAXgene tube for RNA isolation. 
 
A sensitizing dose of 0.4% DPCP ointment will be applied directly to the inner aspect of the 
upper right arm. The site where the dose is applied will be marked and photographed as will the 
extent of scalp disease.  
 
Dose Determination Visit, Day -2 (10 to 14 days post-sensitization) 
Assessments of hair loss, including the SALT score and PAA will be completed. AEs (Appendix 
C) will be monitored and recorded. The sensitization reaction will be recorded by the physician 
on the Sensitization Assessment form using accepted methods (Appendix I, Part I).42 Subjects 
are considered DTH Responders with skin reaction scores ≥ 2+ at the inner arm sensitization site. 
Subjects having skin reaction scores of 0 or 1+ are considered non-responders. In the event the 
subject has a response that is 4+, this will be considered an AE and recorded as such. Reactions 
scoring less than 4+ can still be considered an AE at the investigator’s discretion.  
 
Non-responders (i.e., subjects who do not have a sensitization response) will receive another 
sensitization cycle in the manner previously described. If a subject is found to be a non-
responder through two cycles of sensitization, they will be discontinued from the study. 
 
A concentration of DPCP 0.04 will be applied by the PI or co-PI in the amount of 50 ul per 
concentration to the inner aspect of the upper left thigh. The site where the dose is applied with 
be marked and photographed. The site will be covered with a clear, plastic, waterproof dressing 
such as Tegaderm. The sensitizing test site is to be discontinued 6-8 hours after application by 

Page 11 
 
the subject removing the Tegaderm and cleansing the test site. The subject will report to the 
study site in 72 hours. If the patient has a sufficient reaction (DTH skin reaction score of 1+), this 
will be their treatment dose for the duration of the study and the participant will proceed directly 
to the challenge visit listed below.  
 
- 
If no reaction to the DPCP 0.04% preparation, an additional test sensitizing to an 
alternate site in the inner aspect of the upper left thigh with DPCP 0.08% prepared in 
the same dilutions as previous dosage. The site where the dose is applied with be 
marked and photographed. The sensitizing test is to be discontinued 6-8 hours after 
application by the subject removing the Tegadem and cleansing the test site. The 
subject will report to the study site in 72 hours. If the patient has a sufficient reaction 
[defined as skin reaction score 1+], this will be their treatment dose for the duration of 
the study and the participant will proceed directly to the challenge visit listed below.   
- 
If no reaction to the DPCP 0.08% preparation, an additional test sensitizing to an 
alternate site of the inner aspect of the upper left thigh with DPCP 0.12% prepared in 
the same dilutions as previous dosage. The site where the dose is applied with be 
marked and photographed. The sensitizing test is to be discontinued 6-8 hours after 
application by the subject removing the Tegaderm and cleansing the test site. The 
subject will report to the study site in 72 hours. If the patient has a sufficient reaction 
[defined as skin reaction score 1+], this will be their treatment dose for the duration of 
the study and the participant will proceed directly to the challenge visit listed below.  
- 
If no reaction to the DPCP 0.12% preparation, an additional test sensitizing to an 
alternate site in the inner aspect of the upper left thigh with DPCP 0.2% prepared in 
the same dilutions as previous dosage. The site where the dose is applied with be 
marked and photographed. The sensitizing test is to be discontinued 6-8 hours after 
application by the subject removing the Tegaderm and cleansing the test site. The 
subjects will return to the study site in 72 hours after test applications. If the patient 
has a sufficient reaction [defined as skin reaction score 1+], this will be their 
treatment dose for the duration of the study and the participant will proceed directly 
to the challenge visit listed below.  
- 
If the subject does not have a reaction to the DPCP 0.2%, they will be discontinued 
from the study. If a subject is discontinued from DPCP treatment or study 
participation, they should complete an Early Termination Visit one week after the last 
drug application. 
 
Challenge Visit, Day 0  
 AEs (Appendix C) will be monitored and recorded. 
 
This first treatment of the drug of the determined dose will be applied to the entire scalp (0.75-1 
g of treatment drug) by the PI or co-PI, both board certified dermatologists. Photographs will be 
taken prior to the application of the treatment drug to document extent of hair loss.  
 
Post-First Treatment Sample Collection Visit and Second Treatment, Day 3  
A visit will occur a minimum of 72-96 hours after the first application of drug in which scalp 
biopsy and blood specimens will be collected. Vital signs and AEs (Appendix C) will be 
monitored and recorded. Handling of biopsy and blood specimens will follow the same protocol 

Page 12 
 
as was used at the Sensitization and Baseline Sample Collection Visit, Day -16.  
 
The biopsies taken 3-4 days after application of the first challenge application of DPCP of the 
determined dose will be approximately 2 inches from the first set of biopsies. These biopsies are 
being done to define the initial DTH response to DPCP in the scalp. The expectation is that the 
initial response will differ from a chronic response that becomes more Th1 centered with chronic 
treatment.  
 
The second treatment with study drug will again be applied by the PI or co-PI. Photographs will 
be taken to document extent of hair loss. 
 
Treatment Visits, Twice per Week for Weeks 1-18 
Subjects will return to Dermatology Clinical Research twice per week for application of the 
previously selected dose of DPCP to the whole scalp. Week 18 will only have one treatment 
application visit. Treatment visits must be at least 24 hours apart. The drug will be applied by 
trained a member of the MCRU and/or Dermatology clinical research staff.  AEs will be 
recorded each visit (Appendix C) and will be monitored.  
 
Investigator Assessment Visits (Weeks 4, 8, 12, and 16)  
Every four weeks during treatment, the investigator will attend one of the treatment visits at the 
CRU or Dermatology Clinical Research to perform assessments of hair loss, including the SALT 
score and PAA, in addition to the regular treatments and data collection performed at every 
treatment visit. Photography will also be completed. Vital signs will be recorded and monitored 
at these visits. Pregnancy test will be administered to all women of childbearing potential once 
monthly at these visits. Laboratory tests related to safety monitoring will be drawn at week 12 of 
treatment. 
 
Regrowth Sample Collection Visit 
A visit will occur a minimum of 72-96 hours after a treatment visit in which the physician 
determines the subject has achieved a >50% decrease in hair loss. Scalp biopsy and blood 
specimens will be collected. Vital signs and AEs (Appendix C) will be monitored and recorded. 
Handling of biopsy and blood specimens will follow the same protocol as was used at the 
Sensitization and Baseline Sample Collection Visit, Week 0. These samples will be obtained 
from an area of hair regrowth and ideally, in the same scalp region where the initial biopsy 
samples were obtained. 
 
Follow-up Visit, Week 18  
The follow-up visit will occur a minimum of 72-96 hours after the final application of the drug. 
The post-treatment scalp biopsy and blood specimens will be collected at this time. Vital signs 
and AEs (Appendix C) will be monitored and recorded. Laboratory test related to safety 
monitoring will be drawn at this time. Pregnancy test will be administered to all women of 
childbearing potential. No treatment will be provided. Handling of biopsy and blood specimens 
will follow the same protocol as was used at Week 0. Photography will be completed and 
assessments of hair loss, including the SALT score, the Physician Assessment of Alopecia 
(PAA; Appendix G), and the Subject Hair Loss Assessment (SHLA; Appendix H) will be done 
by the physician or subject. 

Page 13 
 
 
Follow-up Visit, Week 20 
Subjects will return to the CRU or Dermatology Clinical Research 10-14 days after the final 
biopsy collection procedure for suture removal. AEs (Appendix C) will be monitored and 
recorded. A Study Completion Form (Appendix J) will be completed by the investigator or study 
coordinator. 
Early Termination Visit 
If a participant withdraws from the study at any time between the Baseline/Sensitization Visit 
and the final week of treatment, the early termination visit should take place at the time of 
subject withdrawal. AEs (Appendix C) will be monitored and recorded. No treatment will be 
provided. The investigator will perform assessments of hair loss, including the SALT score and 
PAA. The subject will complete the final SHLA (Appendix H). Photography will also be 
completed. A Study Completion Form (Appendix J) will be completed by the investigator or 
study coordinator. 
4. RATIONALE FOR STUDY DESIGN 
The active ingredient used in this study is Diphenylcyclopropenone (DPCP), also known as 
Diphencyprone. The synthesis of DPCP is well established and is readily available from 
commercial sources. DPCP is well accepted in the scientific community as a Delayed type 
Hypersensitivity (DTH) or Contact Hypersensitivity (CHS) Hapten. DPCP has been used for 
decades throughout the world for the treatment of Alopecia Areata (AA), as described in the 
Background section. 
 
Diphenylcyclopropenone is a potent contact sensitizer that has distinct advantages over other 
alternatives. DPCP is not mutagenic in the Ames test and is shelf stable at room temperature.  
DPCP however can break down in the presence of Ultraviolet light and therefore should be 
stored in amber bottles or some other light-blocking container while not in use.  
 
DPCP is not mutagenic in the Ames test, and teratogenicity and organ toxicity could not be 
detected in the hen’s egg test or in the mouse teratogenicity assay. Moreover, DPCP is presumed 
to be not absorbed after topical administration because it is not detected in serum or urine.45 
 
Despite the positive results for treating AA, DPCP has not been FDA approved for the treatment 
because the drug had previously been made in individual pharmacies or clinics and no 
standardized product was available for study. Furthermore, because there was no standardized 
product, many hospitals and clinics are unable to offer this treatment to patients. Hapten 
Pharmaceuticals recently created a standardized formulation of the drug (DPCP Ointment) with 
an IND and studies are needed to demonstrate the effectiveness of this drug in treating AA.  
 
This study includes subjects who have been diagnosed with severe alopecia areata (76-99% hair 
loss) and, in the opinion of the Principal Investigator, are eligible for treatment with DPCP 
Ointment. Subjects with extensive disease are particularly suited to this study as to demonstrate 
the effects of treatment with DPCP Ointment. 

Page 14 
 
4.1 Trial Treatment Descriptions 
Diphencyprone, Diphenylcyclopropenone 
Brand Name: 
Diphenylcyclopropenone Ointment 
Manufacturer:  
Ferndale Laboratories, Inc. 
780 West 8-Mile Road 
Ferndale, MI 48220 
for  
Hapten Pharmaceuticals, LLC 
151 Merriweather Road 
Grosse Pointe, MI 48236 
(313) 300-8454 
Dosage Form 
(1) 0.4% and 0.04% DPCP Ointment 
formulations provided by Hapten 
Pharmaceuticals 
(2) 3 additional strengths of DPCP 0.08%, 
0.12%, 0.2% will be compounded at the U of 
MN Investigational Drug Services Pharmacy 
using the 0.4% and 0.04% DPCP Ointments 
(provided by Hapten Pharmaceuticals); 
treatment dosage will be selected based on 
weakest strength concentration that caused 
minimal reaction  
Color 
Clear 
Route of Administration 
Topical 
Pregnancy Category 
B 
Preservative 
Methyl Paraben 
Propyl Paraben 
Package size 
5 gram tubes of 0.4% DPCP and 0.04% 
ointments (provided for dilution purposes) 
Storage 
Store at room temp 
Avoid excessive heat or exposure to UV light 
Drug Interaction 
None listed 

Page 15 
 
Reported Adverse Events 
Contact dermatitis 
Mild eczema  
Lymph node swelling  
Hyperpigmentation 
Local blistering and swelling 
Pompholyxlike eruptions 
Passive transfer of the sensitizer causing 
eczema distant from the application site and 
regional lymphadenopathy. 
Erythema multiformelike reactions 
Fever 
Palpitations 
Flu-like symptoms 
Headache 
Contraindications 
Allergy or reaction to any component of the 
vehicle formulation. 
Warnings/Precautions 
Diphenylcyclopropenone (DPCP) is a potent 
contact sensitizer. Sensitization of pharmacy, 
medical and nursing staff involved in its 
preparation and use is a potential hazard. 
Gloves should be worn when handling DPCP 
and all used containers, applicators, and 
gloves should be disposed of in a safe manner 
to avoid unintended sensitization to DPCP. 
Potential for hyper- or hypopigmentation, 
especially in patients with dark skin tones. 
 
All medications will be maintained by the Investigational Drug Services (IDS) pharmacy and 
administered within the Clinical Research Unit or Clinical Research Division of the Department 
of Dermatology. Administration will be verified in the numerical identification study file for 
each patient. 
 
Patients will receive all study-related treatments free of charge during the course of the study.   
 
5. SELECTION OF STUDY POPULATION 
This study includes subjects ages 18 and above. Patients of the University of Minnesota 
Dermatology Clinic or affiliated faculty and patients recruited through partnership with the 
National Alopecia Areata Foundation Clinical Trials Network who meet the inclusion/exclusion 
criteria will be given the opportunity to participate in this study. 
5.1 Inclusion Criteria 
Subjects may be included in the study if they meet all of the following criteria: 

Page 16 
 
 
1.  
Subject has clinical diagnosis of extensive alopecia areata (76%-99% involvement 
as determined by SALT score, Appendix B, Part I). 
 
 
2.   
Written informed consent and HIPAA authorization have been obtained.  
 
3.   
Subject is > 18 to years of age.   
 
4. 
Female subjects of childbearing potential have a negative pregnancy test and 
agree to use an acceptable, highly effective method of birth control (i.e., failure 
rate of less than 1% per year) to prevent pregnancy. 
 
5. 
Subject agrees to comply with protocol requirements and attend all required study 
visits and is considered to be a good study subject. 
 
6. 
Subject meets concomitant medication washout requirements. 
 
5.2 Exclusion Criteria 
Subjects will be excluded from the study if they meet any of the following criteria: 
 
1. Subject has <76 or greater than 99% hair loss. 
 
2. Subject is pregnant or lactating. 
 
3. Subject has current controlled or uncontrolled bacterial, viral (with the 
exception of herpes simplex), fungal, atypical, or opportunistic infection(s). 
 
4. Subject has a history of substance abuse within the past five years. 
 
5. Immunosuppression (history of transplantation, chemotherapy, splenectomy, 
HIV). 
 
6. Administration of systemic treatment (e.g., Imuran, biologics) that have an 
immunomodulatory mechanism of action in the preceding 3 months. 
 
7. Previous treatment with DPCP.  
 
8. Application of topical immunomodulating agent in the preceding 6 weeks. 
 
9. Application of topical or intralesional corticosteroids on the scalp within the 
past 6 weeks. 
 
10. Systemic (oral, inhaled, or intravenous) administration of corticosteroid or other 
systemic treatment (i.e., prednisone) with an immunosuppressive mechanism of 
action within the past 3 months. 

Page 17 
 
 
11. Use of light treatments (e.g., PUVA, narrow band UVB) in the preceding 6 
weeks. 
 
12. Use of Anthralin in preceding 6 weeks. 
 
13. Use of minoxidil, topical or oral, in the preceding 4 weeks. 
 
14. Subject is currently or has undergone systemic therapy for malignancy within 
the past five years except for adequately treated Squamous Cell Carcinoma 
(SCC) or Basal Cell Carcinoma (BCC) of the skin. 
 
15. Clinical evidence of secondary skin infection (i.e., folliculitis). 
 
16.  Participation in other therapeutic investigational clinical trials within 4 weeks 
of enrollment.  
 
17. Evidence of anemia, thyroid disease, sarcoidosis or other medical condition that 
could be adversely affected by participating in the study. 
 
18. Subject has any medical condition that, in the judgment of the Investigator, 
would jeopardize the subject’s safety following exposure to the administered 
medications. 
 
Vulnerable Populations: 
 
Population / Group 
Identify whether any of the 
following populations will be 
focus of the research (targeted), 
included, but not necessarily the 
focus or excluded from 
participation in the study. 
Children 
Excluded 
Pregnant women 
Excluded 
Fetuses 
Excluded 
Neonates 
Excluded 
Prisoners 
Excluded. 
Adults lacking capacity to consent 
and/or adults with diminished or 
fluctuating capacity to consent 
Excluded 

Page 18 
 
Non-English speakers 
Excluded 
Those unable to read (illiterate) 
included but not the focus 
Employees of the researcher 
Excluded 
Students of the researcher 
Excluded 
Undervalued or disenfranchised 
social group 
included but not the focus. 
Active members of the military 
(service members), DoD personnel 
(including civilian employees) 
included but not the focus. 
Individual or group that is 
approached for participation in 
research during a stressful 
situation such as emergency room 
setting, childbirth (labor), etc. 
Excluded 
Individual or group that is 
disadvantaged in the distribution 
of social goods and services such 
as income, housing, or healthcare. 
included but not the focus 
Individual or group with a serious 
health condition for which there 
are no satisfactory standard 
treatments. 
Excluded 
Individual or group with a fear of 
negative consequences for not 
participating in the research (e.g. 
institutionalization, deportation, 
disclosure of stigmatizing 
behavior). 
included but not the focus 
Any other circumstance/dynamic 
that could increase vulnerability to 
coercion or exploitation that might 
influence consent to research or 
decision to continue in research. 
included but not the focus. 
 
 
 

Page 19 
 
6. SUBJECT DISPOSITION AND QUALITY ASSURANCE CRITERIA 
6.1 Subject Withdrawal Criteria   
Any subject may discontinue participation in the study for any reason at any time. Subjects who 
withdraw from the study prematurely will complete an Early Termination Visit at the time of 
withdrawal. The procedures to be performed at the Early Termination Visit are specified under 
“Early Termination Visit.” Follow-up evaluations two weeks after the last dose will be obtained, 
if possible. The reason(s) for a subject’s discontinuation from the study will be clearly 
documented in the subject’s medical records, the Case Report Form (See 11.2 Reporting and 
Recording of Data), and on the Study Completion Form (Appendix I).   
 
Patients who withdraw will not be eligible to continue DPCP therapy with the current 
formulation because the drug is only available at this institution on a research basis.  
6.2 Quality Assurance 
To ensure accurate, complete, and reliable data, we will keep records of laboratory tests, clinical 
notes, and the subject’s medical records in the subject’s files as original source documents for the 
study.   
 
The study may be audited by Human Subjects Research Compliance Program of the University 
of Minnesota its representatives and/or regulatory agencies at any time. 
7. TREATMENT OF SUBJECTS 
7.1 Enrollment 
Patients will be recruited by advertising this study to faculty affiliated with the University of 
Minnesota Department of Dermatology and from the investigators’ patients as well as through 
partnership with the National Alopecia Areata Foundation Clinical Trials Network. 
 
Once screening is completed, subject eligibility is confirmed, and written consent obtained, any 
further data, test results, or assessments will be maintained within the patient’s medical file and 
be available to the PI and co-investigators for use in patient care and data collection.  
 
The site contact is: 
Maria Hordinsky, MD 
612-625-1493 
Hordi001@umn.edu 
7.2 Formulation 
The drug substance to be used for this study is: 
 
Diphenylcyclopropenone (Diphencyprone, DPCP) Chemical name: 2,3-diphenylcycloprop2-
en-1-one  
Molecular formula: C15H10O 
Molecular weight: 206.25  

Page 20 
 
Chemical Abstracts (CAS) registry: #886-38-4 
Purity: ≥99% pure crystalline powder. Contains ≤ 0.05% dibromodibenzylketone.  
Commercial Source: Chiracon, Biotechnology Park, 14943 Luckenwalde, Germany 
Contracted by: Carbosynth Ltd, 10 Langley Business Park, Beedon, Berkshire, RG20 8RY, UK  
 
Figure 3.1 Structure of 2,3-diphenylcycloprop-2-en-1-one  
 
Historically, DPCP has been formulated at various strength solutions in acetone. Typically, an 
initial sensitizing dose at a strength of 2-4% in acetone is administered at a distal site (inner arm 
or upper thigh) followed at two weeks by a series of weekly or bi-weekly challenge doses at 
strengths ranging from 0.001 – 0.4% in acetone at the site to be treated. Despite its efficacy, this 
volatile DPCP formulation has faced challenges, mostly due to lack of standardization of 
strength and to side effects caused by over-sensitization. The volatility results in lack of control 
of the actual DPCP concentration on the skin or scalp which most likely contributes to the risk of 
adverse reactions. 
The DPCP Ointment formulation (Samcyprone) being used for the present study is manufactured 
by Ferndale Laboratories, Inc., Michigan under contract to Hapten Pharmaceuticals, LLC, 
Michigan. The drug is manufactured in compliance with FDA’s current Good Manufacturing 
Practices (cGMP) requirements and has been granted Investigational New Drug approval by the 
FDA for indications including warts, melanoma, cutaneous metastasis and neurofibromatosis. 
 
Sensitizing DPCP Ointment (0.4%) and Treatment DPCP Ointment (0.04%) are comprised of the 
excipients shown in the table below. 
 

Page 21 
 
 
 
7.3 Dosage, Administration and Storage 
All subjects will be administered a sensitization dose of 0.05 mL 0.4% DPCP ointment 
formulation topically in the inner aspect of the upper right arm at Day -16, and 0.05 mL of four 
concentrations (0.04, 0.08, 0.12, 0.2%), prepared through dilutions in the ointment vehicle, 
topically on the inner aspect of the left thigh sequentially beginning with the lowest dose 
(0.04%). If no sensitization is obtained with the 0.04% concentration of DPCP, then the subject 
will return and the process will be repeated with the 0.08% concentration and, subsequently, the 
0.12% and the 0.2% if necessary, depending on the reaction of the subject. The weakest strength 
that may cause a minimal reaction (DTH skin reaction score of 1+) after 72 hours will be chosen, 
and 0.75-1 g of that concentration will be applied to the scalp starting at Week 1 and 
administered subsequently twice a week for 18 weeks. 
 
If the research participant has not been sensitized with the first application of 0.4% DPCP a 
second try will be made on the left inner arm. If, again, the patient has not been sensitized, no 
further attempts will be made to sensitize the patient and the patient will be disqualified from the 
study.  
 
Missed Doses 
Patients missing > 2 consecutive treatment visits or 20% of treatment visits in total will not be 
included in subsequent analysis and will not be eligible to continue receiving DPCP therapy 
through our center.  
 
Storage 
The test articles will be stored at Room Temperature (15-25 oC / 59-77 oF) and should not be 
placed in the freezer.  
 

Page 22 
 
7.4 Treatment Schedule 
(See Appendix A) 
Pretreatment Period 
The Demographics and Baseline Assessment Form (Appendix E) and Treatment History Form 
(Appendix F) will be completed at the Screening Visit (and updated at Baseline/Sensitization if 
necessary) to document the preceding 6 weeks before therapy.   
Treatment Periods  
Patients will undergo a single sensitization dose of 0.4% DPCP ointment, a dose 0.05 mL each of 
four concentrations (0.2, 0.12, 0.08, 0.04%) DPCP ointment to determine the treatment dose 
starting with the lowest concentration (0.045%) and the sequentially applying the others by 
increasing concentration until sensitization has been achieved, followed by a 18-week period that 
will involve twice weekly (+/- 2 days) treatment of the scalp with the chosen treatment 
concentration of DPCP ointment.   
Follow-up Period 
Patients will follow-up 72-96 hours after their final DPCP treatment application with a final 
blood and scalp biopsy specimen collection, analysis of safety issues, and hair loss examination. 
The participant must return 10 to 14 days after the biopsy collection for suture removal. 
7.5 Concurrent Medications / Therapies 
All current medications used by the subject throughout the study and all prior medications used 
to treat alopecia areata within 6 weeks prior to the Baseline/Sensitization Visit (Day -16) will be 
recorded in the subject’s medical records, on the Demographics and Baseline Assessment Form 
(Appendix E) and the Treatment History Form (Appendix F).  
7.6 Prohibited Concomitant Medication and Treatment  
 
- Any other therapeutic investigational drugs 
 
- Systemic treatment (e.g., Imuran, or biologics) with an immunomodulatory mechanism 
- Topical immunomodulating agent 
- Oral, intravenous, intralesional (on the scalp), inhaled, or topical (on the scalp) 
corticosteroids 
 
- Systemic immunosuppressants 
- Topical Immunotherapy 
 
- PUVA 
 
- Narrow band UVB 
 
- Anthralin  
 
- Minoxidil, topical or oral 
- Systemic therapy for malignancy within the past five years (except for adequately 
treated SCC or BCC of the skin). 
 
A subject who receives a prohibited concomitant therapy will be asked to withdraw from the 
study.   
 

Page 23 
 
8. EFFICACY ANALYSIS 
8.1 Primary Efficacy Endpoints 
Our primary endpoint of efficacy will be assessed by measuring percentage hair loss at each 
visit. Percentage hair loss will be measured using the Severity of Alopecia Tool (SALT).19-20 The 
percentage of hair loss is determined through a visual examination of four regions of the scalp; 
the percent of terminal hair loss visible in each area is recorded and the percentages are 
aggregated, using SALT scores. 
 
The primary endpoints are as follows, ranked in order of importance: 
 
1. The proportion of subjects who have >75% decrease in their SALT score (SALT75), as 
compared to baseline at the completion of the study or at any time during the study. 
 
2. The proportion of subjects who have >50% decrease in their SALT score (SALT50), as 
compared to baseline at the completion of the study or at any time during the study. 
 
8.2 Exploratory Endpoints 
To identify scalp skin biomarker profiles that characterize a response to DPCP treatment, 
enabling the future potential for predicting individual responses to treatment and the likelihood 
of a sustained response with the opportunity to study blood samples in the future if needed. 
 
Subject assessment of hair loss/growth with the subject hair loss assessment  and investigator 
assessment of the total SALT score.   
 
Gene expression and histologic analysis (immunohistochemistry) on skin biopsy specimens will 
be done. mRNA will be prepared from skin samples stored frozen in RNA Later solution.   
Expression of cytokine genes that define different polar sets of T-cells will be measured: 
interferon-gamma, IL-13, IL-17A, IL-22, IL-9, IL-32; other inflammation associated genes such 
as IL-12, IL-23, IL-1B, and IL-15 will also be measured. mRNAs for a series of negative 
immune regulatory genes, e.g., Foxp3, CTLA4, PD-1, etc. will be measured by RT-PCR. Re-
initiation of hair growth will be assessed by measuring expression of mRNAs encoding hair-
associated keratins and keratin-associated proteins. A subset of patients showing good responses 
of hair re-growth will be selected for whole transcriptome analysis. Pre and post-treatment 
lesional biopsies will have global mRNA profiles established using Affymetrix U133 2.0Plus 
arrays.  From frozen samples in OCT blocks, frozen sections will be prepared that will be stained 
initially with antibodies to T-cells (CD3, CD8), dermal dendritic cells (CD11c), and the 
regulatory marker Foxp3. Once the gene expression analysis is completed, expression of 
negative immune regulatory proteins will be examined, as guided by the genes with highest up-
regulation in post-treatment samples in patients with good hair regrowth. 
 
9. ASSESSMENT OF SAFETY 
Subject safety will be monitored by evaluating all AEs that occur following the first application 
of topical DPCP Ointment and at subsequent treatment visits. 

Page 24 
 
 
If at any time, evidence of clinical and/or laboratory toxicity is noticed, the Investigator will take 
appropriate and prompt measures to elucidate and treat, if necessary, the etiology of the 
condition. The subjects will be followed until the condition resolves or becomes chronic or 
stable.    
9.1 Adverse Events (AE) 
An AE is defined as any untoward medical occurrence (e.g., sign, symptom, disease, syndrome, 
concurrent illness, abnormal laboratory finding) that emerges or worsens relative to pretreatment 
baseline during the study, regardless of the suspected cause. Untoward medical events that occur 
from the time the patient signs the informed consent form to the time the administration of the 
drug starts are not considered adverse events, and will be recorded under medical history. 
 
Adverse events will be recorded as events that are newly appeared, increased in frequency, or 
worsened in severity following application of topical DPCP Ointment. AEs will be recorded 
from the first sensitization does application of the drug until the subject completes the 
study. 
 
The incidence, severity and type of AEs for all treatment-related AEs (defined as definitely, 
probably, or possibly related to the study drug) will be summarized by body system and 
preferred term.  
 
The occurrence of an AE will be determined based on observed or volunteered signs and 
symptoms, as well as changes in the subject’s physical examination and laboratory results.  
 
Patients will be asked at each visit to report any concurrent medications, therapies, illnesses, and 
adverse effects that take place during the study.   
 
All AEs will be evaluated by the Investigator for the seriousness, severity, relationship to topical 
DPCP Ointment, and outcome. Participants will not be compensated monetarily for research-
related injuries. Medical care related to any research-related injuries will be billed to a 
participant’s insurance company.  
 
A non-serious AE is defined as a change from baseline (pre-treatment) in a subject's medical 
health that is not life-threatening, does not require hospitalization, does not prolong a current 
hospitalization, and is not disabling. All non-serious AEs must be reported in the AE form 
(Appendix C) regardless of whether or not they are considered to be related to the study drug. 
An AE occurring will be classified as serious if: 
• It resulted in death (i.e., caused or led to death). 
• It was life-threatening (i.e., placed subject at immediate risk of death; not, however, if it 
could hypothetically have caused death if it were more serious). 
• It required or prolonged inpatient hospitalization (i.e., required at least a 24-hr inpatient 
hospitalization or prolonged a hospitalization beyond expected length of stay; 
hospitalization for elective medical/surgical procedures, scheduled treatments, or routine 
check-ups are not AEs by this criteria). 

Page 25 
 
• It was disabling (i.e., resulted in substantial disruption of the subject’s ability to carry out 
normal life functions). 
• It resulted in a congenital anomaly/birth defect (i.e., adverse outcome in a child or fetus 
of a subject exposed to the molecule or study drug prior to conception or during 
pregnancy). 
• It does not meet any of the above criteria for a serious AE but may jeopardize the subject 
or may require medical or surgical intervention to prevent one of the outcomes listed 
above. 
 
Serious adverse events will be reported to the University of Minnesota IRB, to Hapten 
Pharmaceuticals within 24 hours, and to the FDA.  
9.2 Baseline Medical/Preexisting Conditions 
 
A preexisting condition is one that is present at the start of the study.  A preexisting condition 
will be recorded as an adverse event if the frequency, intensity, or the character of the condition 
worsens during the study period. 
9.3 Procedures for Eliciting, Recording, and Reporting Adverse Events 
AEs may be directly observed or elicited by the Investigator, and/or volunteered by the subjects.  
 
In recording AEs, these guidelines will be observed: 
• Whenever possible, medical terms will not be abbreviated 
• If known, record the diagnosis (disease or syndrome) rather than component signs and 
symptoms unless they are unrelated to an encountered syndrome or disease. 
• Adverse events occurring secondary to other events will be identified by primary cause.   
9.4 Special Reporting Situations 
Fatal or life-threatening events thought to be caused by topical treatment with the investigational 
drug should be immediately reported by telephone.  
 
Death: Event resulting in death should be recorded and reported on the Adverse Event Report 
(See Appendix C) and the Serious Adverse Event Report (Appendix D). 
 
Hospitalization: The illness leading to the surgical or diagnostic procedure will be recorded as 
the AE, not the procedure itself.  
9.5 Post Treatment Follow-Up 
All subjects will be followed for 2-3 weeks after the final application of DPCP Ointment. All 
protocol-defined AEs occurring in treatment periods should be fully evaluated by the 
Investigator and recorded on the appropriate Adverse Event Report (Appendix C) documented in 
the subjects’ files.  
 
Subjects who prematurely discontinue from the study will be encouraged to follow up 6 weeks 
following their last dose. 

Page 26 
 
9.6 Post Adverse Event Follow-Up 
All subjects with AEs will be followed until the event resolves or until participation in the study 
ends. Resolution of those deemed probably related to the administration of the DPCP Ointment 
will be followed until the events resolve or until the Investigator judges the event to be “chronic” 
or “stable.” 
 
9.7 Laboratory Tests 
 
Blood specimens for hematology and chemistry will be collected at the Screening Visit, week 12, 
and at the Follow-up Visit (Week 18 +3d) for all subjects. The following tests will be performed: 
 
Laboratory Tests 
Hematology 
Chemistry 
Hemoglobin 
Hematocrit 
Platelets 
WBC 
Sodium 
Potassium 
Chloride 
Bicarbonate 
BUN/Urea 
Creatinine 
Total protein 
Albumin 
Alkaline phosphatase 
AST (SGOT) 
ALT (SGPT) 
Calcium 
CRP 
 
Sample collection, handling, labeling, and shipping should be conducted following the 
instructions provided by the relevant certified laboratory and the applicable local regulations. 
 
The investigator will review the subjects’ laboratory reports in a timely manner. Screening 
laboratory abnormalities viewed as clinically significant by the investigator that do not exclude 
the subject from study participation will be documented in the subject’s medical record. The 
investigator will complete an appropriate AE form for any new or worsening abnormal test 
results that are identified as clinically significant after baseline. If a subject experiences a 
cardiovascular or hematologic event (including laboratory abnormalities), Grade 2 or higher, the 
study drug will be discontinued. 
 
AEs that may be associated with venipuncture include: 
- 
Pain 
- 
Bruising 
- 
Bleeding at the puncture site 
- 
Fainting 
- 
Inflammation of the vein 

Page 27 
 
10. STATISTICAL METHODS 
10.1 Data to be Analyzed 
Data will be analyzed for efficacy. For primary endpoint analysis, subject data will be analyzed 
up to the point in which compliance is maintained.   
 
Subjects missing > 2 consecutive treatment sessions or 20% of treatment visits in total will not 
be included in subsequent analysis and will not be eligible to continue receiving DPCP therapy 
through our center.  
10.2 Determination of Sample Size 
Statistical considerations 
The primary endpoints ranked below in order of importance are as follows: 
 
1. The proportion of subjects who have ≥ 75% or ≥ 50% decrease in their SALT score 
(SALT75  and SALT50, respectively), as compared to baseline at the completion of the 
study or at any time during the study. 
2. Subject assessment of total scalp hair regrowth or loss using a visual analog scale and 
Investigator assessment of the total SALT score.  
 
The proportion of subjects with ≥75% or ≥50% decrease in the SALT score will be described 
using the fraction achieving the desired decrease and the associated confidence interval, 
computed using the exact Agresti-Coull method. To describe change in SALT score, we will use 
the average change and associated confidence interval.   
 
For a total sample size of 10 participants we anticipate 6 or 8 providing the primary outcomes.  
For changes in the binary outcomes, i.e., proportions having ≥ 75% or ≥ 50% decrease in their 
SALT score, the 95% confidence interval for this proportion is as follows for each possible 
outcome: 
• 6 participants providing data: 0 of 6 meeting the criterion gives an interval of 0 to 0.44;  
1/6, 0 to 0.58;  2/6, 0.09 to 0.70;  3/6, 0.19 to 0.81;  4/6, 0.30 to 0.91;  5/6 0.42 to 1;  and 
6/6, 0.56 to 1. 
• 8 participants providing data:  0/8, 0 to 0.37;  1/8, 0 to 0.49;  2/8, 0.06 to 0.60;  3/8, 0.13 
to 0.70;  4/8, 0.22 to 0.78;  5/8 0.30 to 0.87;  6/8, 0.40 to 0.94;  7/8, 0.51 to 1;  and 8/8, 
0.63 to 1. 
 
 
Exploratory Endpoints 
The primary outcomes for the analysis are levels of T-cell and DC markers. We will compute 
confidence intervals and do paired t-tests for changes in these measures.  These analyses will 
most likely use the logarithms of these measures, as is commonly done; the decision to use or not 
use logarithms will be based on preliminary analyses. 
 
For change in a participant's rating of hair loss using a visual analog scale, the confidence 
interval width is 2 x [97.5th percentile of the t distribution] x the standard deviation of a single 
measurement / square-root (sample size).  For 6 and 8 participants, the 97.5th percentiles are 

Page 28 
 
2.57 and 2.36.  Thus, the respective confidence interval widths are 2.1 standard deviations and 
1.7 standard deviations.  We currently lack information needed to supply a standard deviation 
describing variation between persons in the change in this VAS rating; one of the goals of this 
pilot study is to collect this information. 
 
To estimate power, preliminary data were obtained from a pilot study (Fuentes-Duculan et al. 
Abstract, ISDS, November, 2014). Confidence interval widths are computed as above.  
Assuming the standard deviations are as reported in that abstract, and that the analyses are done 
on the common logarithm scale as in the abstract and analyses are done on the common-log 
scale, the confidence interval widths re-expressed on the original (i.e., non-logarithmically 
transformed) scale are 46-fold and 20-fold differences for 6 and 8 subjects respectively. For tests, 
again assuming the standard deviations are as reported in that aforementioned abstract, for 6 and 
8 participants with data we have 80% power to detect a 14-fold change (using two-tailed alpha = 
0.05).  
 
For the analysis of various biopsy parameters, Dr. Krueger will be supported a statistician in his 
group. 
10.3 Blinding 
No blinding will occur or be necessary. All patients will receive the same treatment plan. 
10.4 Subject Disposition 
The disposition of subjects will describe the number of subjects enrolled, the number of subjects 
treated, and the number of subjects withdrawn prematurely from the study (including the 
reasons).   
10.5 Handling of Dropouts and Missing Data 
The last available SALT scores will be used to determine percent hair regrowth for dropouts and 
subjects who were lost to follow-up.  
 
11. DIRECT ACCESS TO SOURCE DATA/DOCUMENTS 
11.1 Overview 
To decrease variability in this study any manual measurements such as SALT scoring should be 
done as consistently as possible, and be performed by the same individual performing the 
original assessments. Photographs will be taken to verify measurements, if needed. 
11.2 Reporting and Recording of Data 
Case Report Forms (CRF) will be developed and supplied by the University of Minnesota 
Department of Dermatology and will involve laboratory values and other assessments.  
 
A CRF will be completed for each subject enrolled under this protocol. Information collected 
must be verifiable in the source documents. Source documents for this trial will include hospital 
records, clinic records, and any study-specific worksheets. Subject medical records will contain 
reference to the study title and assigned subject identification number. The signed consent form 

Page 29 
 
must be filed with the subject’s medical record.  
 
CRF completion will be kept current to reflect subject status during the course of the trial and 
will be completed in a neat and legible manner to ensure accurate interpretation of data.  
 
Study subjects will be identified by subject ID number, initials, and date of birth.   
 
Results from the study will be available on clinicaltrials.gov. If a participant contacts us wanting 
additional information regarding results, we will provide them with this information.  
12. ETHICAL CONSIDERATIONS 
12.1 Informed Consent 
The informed consent document will be used to explain in simple terms, before the subject is 
entered into the study, the risks and benefits to the subject.  The informed consent document will 
contain a statement that consent is freely given, that the subject is aware of the risks and benefits 
of entering the study, and that the subject is free to withdraw from the study at any time.  
 
The final IRB-approved documents must be provided to the University of Minnesota and 
Sponsors for regulatory purposes. 
 
The Investigator is responsible for seeing that informed consent is obtained from each subject or 
legal representative and for obtaining the appropriate signatures and dates on the informed 
consent document prior to the performance of any protocol procedures and prior to the 
application of topical DPCP ointment formulation, or collection of serum samples and scalp 
biopsy specimens. 
 
As used in this protocol, the term “informed consent” includes all consent given by subjects or 
their legal representatives. 
12.2 Institutional Review 
The University of Minnesota Institutional Review Board will approve the protocol and informed 
consent document and, if appropriate, agree to monitor the conduct of the study and agree to 
review it periodically. The Investigator will provide the IRB with documentation demonstrating 
that the IRB has approved the study before the study begins. 
 
In addition, the Investigator will provide the following documentations: 
• The IRB’s annual re-approval of the protocol 
• The IRB’s approvals of any revisions to the informed consent document or 
amendments to the protocol 
12.3 Ethical Considerations 
The study will be conducted in accordance with the ethical principles stated in the most recent 
version of the Declaration of Helsinki or the applicable guidelines on Good Clinical Practice; 
whichever affords the greater protection to the individual. 

Page 30 
 
12.4 Data Integrity Monitoring 
A CTSI monitor will be used to review the study binders every 6 months while we are actively 
recruiting and enrolling participants. The objective of the monitor will be to ensure completion 
of forms including adverse event forms, reporting, and to assess for other protocol deviations.  
The monitor will be expected to write a report of major and minor protocol deviations and 
provide to the study staff.   
 
 
15. FINANCING AND INSURANCE 
 
Considerations will be provided by the University of Minnesota Department of Dermatology 
fund for Graduate Research and the National Alopecia Areata Foundation.  
 
 
16. REFERENCES 
 
1.  
Alkhalifah A, Alsantali A, Wang E, McElwee KJ, Shapiro J. Alopecia areata update. J Am 
Acad Dermatol. 2010;62(2):177-188. doi:10.1016/j.jaad.2009.10.032. 
2.  
Safavi KH, Muller SA, Suman VJ, Moshell AN, Melton LJ. Incidence of alopecia areata 
in Olmsted County, Minnesota, 1975 through 1989. Mayo Clin Proc. 1995;70(7):628-633. 
doi:10.1016/S0025-6196(11)63913-X. 
3.  
Al-Mutairi N, Eldin ON. Clinical profile and impact on quality of life: seven years 
experience with patients of alopecia areata. Indian J Dermatol Venereol Leprol. 
2011;77(4):489-493. doi:10.4103/0378-6323.82411. 
4.  
Firooz A, Firoozabadi MR, Ghazisaidi B, Dowlati Y. Concepts of patients with alopecia 
areata about their disease. BMC Dermatol. 2005;5(1):1. doi:10.1186/1471-5945-5-1. 
5.  
Williamson D, Gonzalez M, Finlay AY. The effect of hair loss on quality of life. J Eur 
Acad Dermatol Venereol. 2001;15(2):137-139. 
http://www.ncbi.nlm.nih.gov/pubmed/11495520. 
6.  
Colón EA, Popkin MK, Callies AL, Dessert NJ, Hordinsky MK. Lifetime prevalence of 
psychiatric disorders in patients with alopecia areata. Compr Psychiatry. 1991;32(3):245-
251. http://www.ncbi.nlm.nih.gov/pubmed/1884604. 
7.  
Ruiz-Doblado S, Carrizosa A, García-Hernández MJ. Alopecia areata: psychiatric 
comorbidity and adjustment to illness. Int J Dermatol. 2003;42(6):434-437. 
http://www.ncbi.nlm.nih.gov/pubmed/12786868. 
8.  
Jackow C, Puffer N, Hordinsky M, Nelson J, Tarrand J, Duvic M. Alopecia areata and 
cytomegalovirus infection in twins: genes versus environment? J Am Acad Dermatol. 
1998;38(3):418-425. http://www.ncbi.nlm.nih.gov/pubmed/9520023. 
9.  
Blaumeiser B, van der Goot I, Fimmers R, et al. Familial aggregation of alopecia areata. J 
Am Acad Dermatol. 2006;54(4):627-632. doi:10.1016/j.jaad.2005.12.007. 
10.  
Goh C, Finkel M, Christos PJ, Sinha AA. Profile of 513 patients with alopecia areata: 
associations of disease subtypes with atopy, autoimmune disease and positive family 
history. J Eur Acad Dermatol Venereol. 2006;20(9):1055-1060. doi:10.1111/j.1468-
3083.2006.01676.x. 
11.  
Petukhova L, Duvic M, Hordinsky M, et al. Genome-wide association study in alopecia 

Page 31 
 
areata implicates both innate and adaptive immunity. Nature. 2010;466(7302):113-117. 
doi:10.1038/nature09114. 
12.  
Gilhar A, Ullmann Y, Berkutzki T, Assy B, Kalish RS. Autoimmune hair loss (alopecia 
areata) transferred by T lymphocytes to human scalp explants on SCID mice. J Clin 
Invest. 1998;101(1):62-67. doi:10.1172/JCI551. 
13.  
Cetin ED, Şavk E, Uslu M, Eskin M, Karul A. Investigation of the Inflammatory 
Mechanisms in Alopecia Areata. Am J Dermatopathol. 2009;31(1):53-60. 
doi:10.1097/DAD.0b013e318185a66e. 
14.  
Paus R, Ito N, Takigawa M, Ito T. The hair follicle and immune privilege. J Investig 
Dermatology Symp Proc. 2003;8(2):188-194. doi:10.1046/j.1087-0024.2003.00807.x. 
15.  
Kang H, Wu W-Y, Lo BKK, et al. Hair follicles from alopecia areata patients exhibit 
alterations in immune privilege-associated gene expression in advance of hair loss. J 
Invest Dermatol. 2010;130(11):2677-2680. doi:10.1038/jid.2010.180. 
16.  
Delamere FM, Sladden MM, Dobbins HM, Leonardi-Bee J. Interventions for alopecia 
areata. Cochrane Database Syst Rev. 2008;(2):CD004413. 
doi:10.1002/14651858.CD004413.pub2. 
17.  
Alkhalifah A, Alsantali A, Wang E, McElwee KJ, Shapiro J. Alopecia areata update: part 
II. Treatment. J Am Acad Dermatol. 2010;62(2):191-202, NaN-4. 
doi:10.1016/j.jaad.2009.10.031. 
18.  
MacDonald Hull SP, Wood ML, Hutchinson PE, Sladden M, Messenger AG, British 
Association of Dermatologists. Guidelines for the management of alopecia areata. Br J 
Dermatol. 2003;149(4):692-699. http://www.ncbi.nlm.nih.gov/pubmed/14616359. 
19.  
Kennedy Crispin M, Ko JM, Craiglow BG, et al. Safety and efficacy of the JAK inhibitor 
tofacitinib citrate in patients with alopecia areata. JCI Insight. 2016;1(15). 
doi:10.1172/jci.insight.89776. 
20.  
Olsen E, Hordinsky M, McDonald-Hull S, et al. Alopecia areata investigational 
assessment guidelines. National Alopecia Areata Foundation. J Am Acad Dermatol. 
1999;40(2 Pt 1):242-246. http://www.ncbi.nlm.nih.gov/pubmed/10025752. 
21.  
Olsen EA, Hordinsky MK, Price VH, et al. Alopecia areata investigational assessment 
guidelines--Part II. National Alopecia Areata Foundation. J Am Acad Dermatol. 
2004;51(3):440-447. doi:10.1016/j.jaad.2003.09.032. 
22.  
Happle R, Klein HM, Macher E. Topical immunotherapy changes the composition of the 
peribulbar infiltrate in alopecia areata. Arch Dermatol Res. 1986;278(3):214-218. 
http://www.ncbi.nlm.nih.gov/pubmed/2873796. 
23.  
Herbst V, Zöller M, Kissling S, Wenzel E, Stutz N, Freyschmidt-Paul P. 
Diphenylcyclopropenone treatment of alopecia areata induces apoptosis of perifollicular 
lymphocytes. Eur J Dermatology. 2006;16(5):537-542. 
http://www.ncbi.nlm.nih.gov/pubmed/17101475. 
24.  
Happle R, Echternacht K. [Alopecia areata: successful half-side treatment with DNCB]. Z 
Hautkr. 1977;52(22):1129-1134. http://www.ncbi.nlm.nih.gov/pubmed/595747. 
25.  
Happle R. The potential hazards of dinitrochlorobenzene. Arch Dermatol. 
1985;121(3):330-332. http://www.ncbi.nlm.nih.gov/pubmed/3977355. 
26.  
Happle R, Hausen BM, Wiesner-Menzel L. Diphencyprone in the treatment of alopecia 
areata. Acta Derm Venereol. 1983;63(1):49-52. 
http://www.ncbi.nlm.nih.gov/pubmed/6191489. 
27.  
Fiedler VC. Alopecia Areata: A Review of Therapy, Efficacy, Safety, and Mechanism. 

Page 32 
 
Arch Dermatol. 1992;128(11):1519. doi:10.1001/archderm.1992.01680210097016. 
28.  
van der Steen PH, van Baar HM, Happle R, Boezeman JB, Perret CM. Prognostic factors 
in the treatment of alopecia areata with diphenylcyclopropenone. J Am Acad Dermatol. 
1991;24(2 Pt 1):227-230. http://www.ncbi.nlm.nih.gov/pubmed/2007667. 
29.  
Rokhsar CK, Shupack JL, Vafai JJ, Washenik K. Efficacy of topical sensitizers in the 
treatment of alopecia areata. J Am Acad Dermatol. 1998;39(5 Pt 1):751-761. 
http://www.ncbi.nlm.nih.gov/pubmed/9810892. 
30.  
Gordon PM, Aldrige RD, McVittie E, Hunter JA. Topical diphencyprone for alopecia 
areata: evaluation of 48 cases after 30 months’ follow-up. Br J Dermatol. 
1996;134(5):869-871. http://www.ncbi.nlm.nih.gov/pubmed/8736327. 
31.  
Hordinsky MK, Bertin A, Levin A. Treatment of alopecia areata. In: Maibach H, Gorouhi 
F, eds. Evidence Based Dermatology. 2nd ed. Sheldon, CT: People’s Medical Publishing 
House - USA; 2011:283-300. 
32.  
Pardasani AG, Turner E, McMichael AJ. Squaric acid dibutylester: indications for use and 
efficacy in alopecia areata. Arch Dermatol. 2001;137(7):970-972. 
http://www.ncbi.nlm.nih.gov/pubmed/11453831. 
33.  
Ajith C, Gupta S, Kanwar AJ. Efficacy and safety of the topical sensitizer squaric acid 
dibutyl ester in Alopecia areata and factors influencing the outcome. J Drugs 
Dermatology. 2006;5(3):262-266. http://www.ncbi.nlm.nih.gov/pubmed/16573260. 
34.  
Avgerinou G, Gregoriou S, Rigopoulos D, Stratigos A, Kalogeromitros D, Katsambas A. 
Alopecia areata: topical immunotherapy treatment with diphencyprone. J Eur Acad 
Dermatol Venereol. 2008;22(3):320-323. doi:10.1111/j.1468-3083.2007.02411.x. 
35.  
Luk NM, Chiu LS, Lee KC, et al. Efficacy and safety of diphenylcyclopropenone among 
Chinese patients with steroid resistant and extensive alopecia areata. J Eur Acad Dermatol 
Venereol. 2013;27(3):e400-5. doi:10.1111/jdv.12009. 
36.  
Freyschmidt-Paul P, Happle R, McElwee KJ, Hoffmann R. Alopecia areata: treatment of 
today and tomorrow. J Investig Dermatology Symp Proc. 2003;8(1):12-17. 
doi:10.1046/j.1523-1747.2003.12165.x. 
37.  
Czarnowicki T, Malajian D, Shemer A, et al. Skin-homing and systemic T-cell subsets 
show higher activation in atopic dermatitis versus psoriasis. J Allergy Cinical Immunol. 
2015;136(1):208-211. doi:10.1016/j.jaci.2015.03.032. 
38.  
Krueger JG, Fretzin S, Suárez-Fariñas M, et al. IL-17A is essential for cell activation and 
inflammatory gene circuits in subjects with psoriasis. J Allergy Cinical Immunol. 
2012;130(1):145-54.e9. doi:10.1016/j.jaci.2012.04.024. 
39.  
Suárez-Fariñas M, Ungar B, Noda S, et al. Alopecia areata profiling shows TH1, TH2, and 
IL-23 cytokine activation without parallel TH17/TH22 skewing. J Allergy Clin Immunol. 
2015;136(5):1277-1287. doi:10.1016/j.jaci.2015.06.032. 
40.  
Kim J, Oh C-H, Jeon J, et al. Molecular Phenotyping Small (Asian) versus Large 
(Western) Plaque Psoriasis Shows Common Activation of IL-17 Pathway Genes but 
Different Regulatory Gene Sets. J Invest Dermatol. 2016;136(1):161-172. 
doi:10.1038/JID.2015.378. 
41.  
Levis WR, Holzer AM, Leonard LK. Topical diphenylcyclopropenone as a measure of 
immune competence in HIV-seropositive subjects. J Drugs Dermatology. 2006;5(9):853-
858. http://www.ncbi.nlm.nih.gov/pubmed/17039650. 
42.  
Gulati N, Suárez-Fariñas M, Fuentes-Duculan J, et al. Molecular characterization of 
human skin response to diphencyprone at peak and resolution phases: therapeutic insights. 

Page 33 
 
J Invest Dermatol. 2014;134(10):2531-2540. doi:10.1038/jid.2014.196. 
43.  
Gulati N, Carvajal RD, Postow MA, Wolchok JD, Krueger JG. Definite regression of 
cutaneous melanoma metastases upon addition of topical contact sensitizer diphencyprone 
to immune checkpoint inhibitor treatment. Exp Dermatol. 2016;25(7):553-554. 
doi:10.1111/exd.13030. 
44.  
Gulati N, Garcet S, Fuentes-Duculan J, et al. Molecular Profiling of Immune Activation 
Associated with Regression of Melanoma Metastases Induced by Diphencyprone. Vol 
136.; 2016. doi:10.1016/j.jid.2016.06.611. 
45.  
Berth-Jones J, Mc Burney A, Hutchinson PE. Diphencyprone is not detectable in serum or 
urine following topical application. Acta Derm Venereol. 1994;74(4):312-313. 
http://www.ncbi.nlm.nih.gov/pubmed/7976097. 
Add reference 
 

Page 34 
 
 
17. Appendices 
 
 
An Open Label Study to Evaluate DPCP Ointment for the Treatment of Alopecia Areata 
 
Appendices: 
A. Flowchart of Study Procedures 
B. Instructions for Severity of Alopecia Tool Assessments 
C. Adverse Events Report Log 
D. Serious Adverse Event Report Form 
E. Demographics and Baseline Assessment 
F. Treatment History Form 
G. Physician Assessment of Alopecia 
H. Subject Hair Loss Assessment 
I. Sensitization Assessment 
J. Study Completion Form

Page 35 
 
APPENDIX A: Flowchart of Study Procedures 
Week(s) 
>= -2 
-1 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
17 
18 
20 
Visit(s) 
1a 
2 
3n 4 
5 6-7b 8-9 10-
11 12 13 14-
15 
16-
17 
18-
19 20 21 22-
23 24-25 26-
27 28 29 30-
31 32-33 34-
35 36 37 38-
39 40 41 42 
Study Event 
Screening 
Baseline, 
Sensitization 
Dose 
Challenge 
Treatment 
Assessment 
Treatment 
Assessment 
Treatment 
Assessment 
Treatment 
Assessment 
Treatment 
Assessment 
Suture 
Removal
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Review of Inclusion and 
Exclusion Criteria 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Document Interim Medical 
Hx/Adverse Experiences 
 
X 
X X X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Vital Signs 
X 
X 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
X 
 
SALT Score 
 
X 
X 
X 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
X 
 
Physician Assessment of 
Alopecia 
 
X 
X 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
X 
 
Subject Hair Loss 
Assessments 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Skin Biopsiesc 
 
X 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Xd 
 
Phlebotomye 
 
X 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Xd 
 
Suture Removalf 
 
 
 
X 
 
Xg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Xh 
Photography 
 
X 
X 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
X 
 
Drug Application 
 
Xi 
Xj X X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
Assessment of Sensitization 
 
 
X X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographic & Baseline 
Assessment 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Laboratory Testsk,l 
Xk,l 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Xl 
 
 
 
 
 
 
 
Xl 
 
Pregnancy Testm 
X 
X 
 
 
 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
 
X 
 
 
X 
 
 
a Screening visit can but does not have to occur concurrently with sensitization/baseline. 
b Subjects treatment visits twice per week for 18 weeks after the challenge visit. 

Page 36 
 
c Scalp skin biopsies will be performed at baseline, 3 days after the first treatment administration, at the time of hair regrowth (if >=50%) OR after 
the final treatment administration  
d 3rd biopsy and phlebotomy collection will occur at Visit 41 OR a prior date at the time of regrowth (if >=50%) 
e Whole blood, serum collection 
f Sutures will be removed 10-14 days after the biopsy procedure 
g Of Visits 6-7, suture removal will only take place on Visit 6 
h Suture removal will occur at Visit 41 OR at a prior visit 10-14 days after the time of regrowth (if >=50%) 
i Drug sensitization procedure 
j Dose determination procedure 
k Pre-treatment labs include TSH, HgB, and Vitamin D 
l Safety monitoring labs include hemoglobin, hematocrit, platelets, WBC, sodium, potassium, chloride, bicarbonate, BUN/urea, total protein, 
albumin, alkaline phosphate, AST (SGOT), ALT (SGPT), calcium, CRP 
m Women of child bearing potential only 
n See following schedule of events for dose determination visit(s): 
 
 
Visit 
Procedure 
31 32 33 34 
Adverse events 
X X X X 
Assessment of hair loss 
X  
 
 
Photographs 
X X X X 
DPCP ointment – 0.04% 
X  
 
 
DPCP ointment – 0.08% 
 
X  
 
DPCP ointment – 0.12% 
 
 
X  
DPCP ointment – 0.2% 
 
 
 
X 
 
1: DPCP ointment 0.04% concentration is the starting concentration for all participants  
2: DPCP ointment 0.08% will only be applied if participant does not have a strong enough response to the 0.005% concentration of DPCP. Thus, this visit will begin with an 
assessment of the sensitization reaction to the 0.005% dose, If there is not a strong enough response, 0.01% concentration DPCP will be applied to a different area of the inner 
upper left thigh in the same manner as the 0.005% dose. If the participant is a responder to the 0.005% DPCP, patient will proceed to the challenge visit (visit 4) as described 
above.  
3: DPCP ointment 0.12% will only be applied if participant does not have a strong enough response to the 0.01% concentration of DPCP. Thus, this visit will begin with an 
assessment of the sensitization reaction to the 0.01% dose, If there is not a strong enough response, 0.05% concentration DPCP will be applied to a different area of the inner 
upper left thigh in the same manner as the 0.01% dose. If the participant is a responder to the 0.01% DPCP, patient will proceed to the challenge visit (visit 4) as described above.  
4: DPCP ointment 0.2% will only be applied if participant does not have a strong enough response to the 0.05% concentration of DPCP. Thus, this visit will begin with an 
assessment of the sensitization reaction to the 0.05% dose, If there is not a strong enough response, 0.1% concentration DPCP will be applied to a different area of the inner upper 
left thigh in the same manner as the 0.05% dose. If the participant is a responder to the 0.05% DPCP, patient will proceed to the challenge visit (visit 4) as described above.  
 

Page 37 
 
APPENDIX B: Instructions for Severity OF Alopecia Tool Assessments 
Part I: The Severity of Alopecia Tool (SALT)  
 
 
Overview 
 
The SALT score is a global alopecia areata severity score based on scalp hair loss, as it provides 
a measure of the percentage of total scalp alopecia. The SALT score is calculated by visually 
examining four discrete areas of the scalp, measuring the percent of terminal hair loss in each 
area and then totaling the results. (See below) 
 
The SALT score will be measured to the nearest 1%, following the instructions below, at 
screening to determine study eligibility and at all following appointments as one measure of the 
extent of hair loss. 
 
Evaluator Requirements 
 
Dr. Maria Hordinsky and Dr. Ronda Farah will be the primary evaluators and will be performing 
all percentage hair loss assessments for all individual subjects throughout the trial. 
 
Determining SALT score 
 
To determine the SALT score, four main scalp areas are assessed:  the left side, the right side, the 
top and the back.  These areas correspond, respectively, to 18%, 18%, 40%, and 24% of the total 
scalp surface area (see Figure A, “Visual Aid for Determining SALT Score”).  
 
Figure A: Visual Aid for Determining SALT Score 
 
 
Reference:  Figure 1. “Alopecia Areata Investigational Assessment Guidelines –Part II” (Olsen, 
et. al. 2004). 
 
 

Page 38 
 
Determining SALT Score (cont) 
 
The subject’s SALT score will be determined as follows: 
1. For each of the four main scalp surface areas, the percentage of missing hair will be 
documented in Row 1. 
2. The percentages recorded in Row 1 will be multiplied by the percentages in Row 2 of the 
same column (Areas as % of Total Scalp Area).  The results will be entered in the 
appropriate columns of Row 3. 
3. The four results in Row 3 will be summed to get the percentage hair loss (Row 4).  
 
Sample Worksheet for Determining SALT Score 
 
Row 
Parameter 
Left 
Right 
Top 
Back 
1 
Enter % Hair loss 
for Each Scalp 
Surface Area 
 
 
 
 
2 
Area as % of 
Total Scalp 
Surface 
X 0.18 
X 0.18 
X0.4 
X 0.24 
3 
Multiply Row 1 x 
Row 2 
 
 
 
 
4 
Total % Hair 
Loss (Sum the 
Products from 
Row 3) 
 
 
50% scalp hair regrowth based on SALT score can be expressed as SALT50 
75% scalp hair regrowth based on SALT score can be expressed as SALT25 

 Page 39 
  
APPENDIX C: Adverse Event Report Log 
ADVERSE EVENTS 
Has the Subject experienced any Adverse Events since receiving the study treatment? 
 Yes      No 
Description of Event 
Serious 
Y/N* 
DLT 
Y/N 
Related to 
Treatment 
Y/N 
Severity** 
Onset Date 
(dd/mmm/yyyy) 
Resolution 
Date or 
Ongoing 
(dd/mmm/yyyy) 
Relationship 
To Study 
Drugǂ 
Action 
Taken 
Regarding 
Study Drugǂǂ 
Action 
Taken to 
Treat 
Adverse 
Event 
^ (if 3,4 or 
5, specify) 
Outcome^^ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*If yes, complete an SAE CRF. 
**Severity 
ǂRelationship 
ǂǂAction Taken Regarding Study Drug 
^Action Taken to Treat Adverse Event 
^^Outcome 
1 Mild 
2 Moderate 
3 Severe 
4 Life-
Threatening 
1 Definite 
2 Probable 
3 Possible 
4 Not Related 
1 None – N/A 
2 Dose Discontinued 
 
1 None 
2 Concomitant Medication 
3 Blood Product 
4 Medical or Surgical Procedure 
5 Other Intervention 
1 Continuing 
2 Resolved 
3 Lost to Follow-up 
4 Unknown 
5 Resolved with sequelae 
6 Fatal 
Investigator’s Signature:___________________  Date 
/
/20
 (dd/mmm/yyyy)   

 Page 40 
  
APPENDIX D: Serious Adverse Event Report 
(Fax completed form to 888-929-9821) 
Protocol No. 
                 
Site No. 
 
Principal Investigator 
 
Date Form Completed  
 
 
□   Initial Report            □   Follow-Up Report #______       □   Final 
Report 
Reporter Name 
 
 
Title 
Telephone Number 
 
Serious Adverse Event 
 
 
 
Onset Date 
Resolution Date 
 
□  or Ongoing 
 
Subject ID  
Gender 
Race 
Age 
Height 
Weight 
 
 
 
 
 
 
 
 
 
Study Design 
              □  Open-Label                □  Double-Blind                □  Other (specify): Single Blind 
________________________________ 
Investigational Drug 
DPCP (SamcyproneTM) 
Dose Date  
Dose 
Route of Administration 
Topical 
Prescription #  
 
 
 
 
 
Criteria for Seriousness   (check all that apply) 
Action Taken For Event 
□  Death 
□  Life Threatening 
□  Inpatient Hospitalization * 
□  Prolongation of Existing Hospitalization * 
□  Persistent or Significant Disability / Incapacity 
□  Congenital Anomaly / Birth Defect 
□  Required Intervention to Prevent Permanent Damage 
□  Other (specify) __________________________________ 
□  None 
□  Investigational Drug Dose Discontinued 
□  Required Therapy (specify on page 2) 
 
* Dates of Hospitalization 
Admission Date 
 
 
Discharge Date 
 
 
 
Event Status at Time of This Report 
Grade of Event 
(CTCAE) 
Investigator Assessment of 
Relationship to Investigational 
Drug 
Was Investigational 
Drug Resumed? 
□  Recovered/Resolved  
□  Not Recovered/Resolved 
□  Recovered/Resolved with 
      Sequelae  
□  Died 
□  Unknown 
□  1 
□  2 
□  3 
□  4 
□  5 
□  Related 
□  Not Related 
□  Yes 
□  No 
□  N/A 
 

 Page 41 
  
 
 
 
 
 
 
 
Protocol No. 
 
Site No. 
 
Subject ID 
 
 
Known Medications at the Time of the Event 
1) 
Exclude those used to treat the event 
2) 
A copy of the Concomitant Medication CRF may be attached in place of completing this section   
Drug (brand name if known) 
Total Daily Dose 
Units 
Route 
Indication 
Start Date 
Stop Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medications Used to Treat the Event 
Drug (brand name if known) 
Total Daily Dose 
Units 
Route 
Indication 
Start Date 
Stop Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Relevant Tests / Laboratory Data 
1) A copy of the lab reports may be attached in place of completing this section 
Clinical Laboratory Test 
Normal Range 
Value /  Date 
Value /  Date 
Value /  Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Relevant Background Information 
1) Include medical history and demographic data, including pre-existing medical conditions 
 
 
 
 
 
 
 
 
 
 
 
 
 

 Page 42 
  
 
 
 
 
Protocol No. 
 
Site No. 
 
Subject ID 
 
 
 
Narrative 
1) 
Please include any other important information that is not included on the form. 
2) 
Supplemental information or supporting documents may be attached. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Investigator’s Signature 
 
 
Date 
 
 
Please Email copy of this completed form to Hapten 
Pharmaceuticals: michaelburnsus@aol.com 
 
 
 
 

 Page 43 
  
APPENDIX E: Demographics & Baseline Assessment  
 
(Visit 1, Date of visit: ____/____/____) 
 
1.  Subject Initials:  ___ ___ ___ 
   2. Subject study number (ID #): ___ ___ ___ 
 
3.  Date of Birth: ___/ ___/___  
   4. Gender:     Male      Female 
 
5.   Height?  __ ft. __  in. /  __m. __ cm.    6. Weight?  
______lb. / _____ kg. 
 
7. Blood Pressure:  _________ mmHg    8.  Pulse _________ bpm 
 
5. If female, urine pregnancy test: 
      Positive        Negative 
 
6. Ethnicity: 
 Hispanic/Latino 
 
 Not Hispanic/Latino 
 
7. Race: 
 Caucasian 
        Black/African American          American Indian 
 
 
 Asian 
 
 Pacific Islander                  Multi-racial 
 
 
 
 
Other: __________________ 
 
   
            Refused 
 
8. Fitzpatrick Skin Type: 
 
 I – Always burns, never tans.  
         IV – Rarely burns, tans well 
 
 II – Always burns, rarely tans 
         V – Rarely burns, always tans 
 
 III – Sometimes burns, gradually tans   
 VI – Never burns, always tans 
 
9. Significant or Recent Medical History:  
 
Condition/Illness/Surgery/Hospitalizations 
Condition Status (check if condition is 
ongoing or resolved) 
Ongoing 
Resolved 
 
 
 
 
 
 
 
 
 
 
 
 
 Subjects denies any significant medical history, hospitalizations or surgeries 
  
10. Medications Used in Past 6 weeks (Including Current):  
 
Medication 
 
Dosage 
 
Frequency 
 
Indication 
 
Date Started 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  Subject denies any medication or OTC regimen 
 

 Page 44 
  
APPENDIX F: Treatment History Form  
 
(Visit 1, Date of visit: ____/____/____) 
Of these commonly used therapies for alopecia areata, check all that you use now and have used in the past in the 
following table:   
 
Therapies: 
Current 
Treatment 
And Dose 
Past 
Therapies 
used 
Check if Hair 
Re-growth 
achieved 
Any Side effects of therapy? 
None 
_________ 
          
          
 
 
Systemic Steroids  “cortisone” 
      Oral (prednisone, medrol) 
      Intramuscular 
 
 
_________ 
_________ 
 
 
          
          
 
 
          
          
 
 
No Yes  ________________ 
No Yes  ________________ 
 
Intralesional steroids  
 
_________ 
 
          
 
          
 
No Yes  ________________ 
Topical steroids 
      Gel or mousse 
      Lotion, Cream or ointment 
 
_________ 
_______ 
 
 
          
          
 
          
          
 
No Yes  ________________ 
No Yes  ________________ 
Topical Immunotherapy 
      FK506 (protopic, tacrolimus) 
      Cyclosporine 
      Aldara  
      Elidel 
      Other ________________ 
 
_________ 
________ 
_________ 
_________ 
_________ 
 
          
          
          
          
          
 
          
          
          
          
          
 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
Oral Immunotherapy 
      FK506 (tacrolimus) 
      Cyclosporin 
      Other _______________ 
 
_________ 
_________ 
_________ 
 
          
          
          
 
          
          
          
 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
Phototherapy  
      NB UVB 
      PUVA – systemic 
      PUVA – topical 
      Excimer laser LLLT  
 
_________ 
_________ 
_________ 
_________ 
 
          
          
          
          
 
          
          
          
          
 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
Contact Sensitizer 
      DNCB Dinitrochlorobenzene 
      Diphencyprone 
      Squaric acid dibutylester 
 
_________ 
_________ 
_________ 
 
          
          
          
 
          
          
          
 
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
Retinoids  (Vitamin A) 
      Topical _______________   
      Oral   ________________ 
 
_________ 
_________ 
 
 
          
          
 
          
          
 
No Yes  ________________ 
No Yes  ________________ 
Topical Rogaine/Minoxidil 
      2% 
      5% 
 
Topical Anthraline/Dithrocream 
 
_________ 
_________ 
 
_________ 
 
 
          
          
          
          
 
 
          
          
          
          
 
 
No Yes  ________________ 
No Yes  ________________ 
 
No Yes  ________________ 
Other______________________ 
Other______________________ 
Other______________________ 
 
_________ 
_________ 
_________ 
          
          
          
          
          
          
No Yes  ________________ 
No Yes  ________________ 
No Yes  ________________ 
 
 
 

 Page 45 
  
 
APPENDIX G: Physician Assessment of Alopecia 
 
The Physician Assessment of Alopecia includes body hair assessment, nail involvement assessment, duration 
of current episode of scalp hair loss pattern of scalp hair loss, and SALT scoring assessment. 
 
The proportion of scalp involvement is determined by dividing the scalp into 4 quadrants and estimating the 
scalp surface that all the alopecic areas would occupy if placed together. The following groups will be used: 
 
 
 
S0 (no scalp hair loss)  
   S3 (51-75% hair loss) 
 
 
 
S1 (<=25% hair loss)   
   S4 (76-99% hair loss) 
 
 
 
S2 (26-50% hair loss)    S5 (100% hair loss) 
 
 
Other areas of alopecia or involvement by alopecia areata may be noted: 
 
B. Extent of Body Hair Loss on the exam today: 
 
 
B0 (no body hair loss) 
 
B2 (100% body hair loss) 
 
 
B1 (some body hair loss): 
 
 
 
  Eyelashes   Arms 
                 Eyebrows 
  Trunk 
 
 
  Beard 
  Pubic area 
 
 
  Axilla 
  Legs 
 
N. Degree of Nail Involvement: 
 
 
 
 N0 (no nail involvement)     
 
 
 
 N1 (some nail involvement) 
 
 
 
 N2 (all nails) 20 nail dystrophy 
 
 
 
 
 
 
Area:   Fingernails  Toenails 
 Both 
 
 
 
Type:   Pitting  Onycholysis  Onychomycosis  Dystrophy 
 
Global assessment: overall improvement. This takes into account extent and density of regrowth by the SALT 
scoring system. First note regrowth, then categorize by: 
  A0 = no change or further loss 
  A1 = 1-24% regrowth 
  A2 = 25-49% regrowth 
  A3 = 50-74% regrowth 
  A4 = 75-99% regrowth 
  A5 = 100% regrowth 
 
 

 Page 46 
  
APPENDIX H: Subject Hair Loss Assessment 
 
Visit Date: ___/___/___ 
Week: _______ 
 
Instructions:  To be completed by the subject. Grade your perception of the extent of your hair loss for your 
scalp and the rest of your body. Check only one box per row. 
 
 
Region 
Severe 
Moderate 
to Severe Moderate 
Mild to 
Moderate 
Mild 
Trace 
None 
Scalp 
Non-Scalp 
 
 
Subject Signature: __________________________    Date: ___________________  
 
 
 
 
 

 Page 47 
  
APPENDIX I: Sensitization Assessment 
 
Part I: Sensitizing Dose 
 
Immunotherapeutic Response Skin Reaction Scale 
Score  
Observation 
0 
 
No skin reaction 
1+  
 
Erythema only  
2+ 
 
Erythema and cutaneous induration  
3+ 
 
Erythema, papules and small vesicles  
4+ 
 
Large vesicles, bullae, and severe local reaction besides erythema 
First Attempt: (Visit ___, Date: ___/___/___) 
 
1.  Did skin sensitization occur?     Yes 
 No 
 
SKIN 
REACTION 
SCORE 
0 
1+ 
2+ 
3+ 
4+ 
 
 
If no, was another sensitization dose applied?       Yes 
 No 
 
2. Did the subject experience any adverse reactions to the sensitizing dose? 
     Yes          No   
    * If yes, note reaction and file AE form:   ______________________________________ 
____________________________________________________________________________ 
____________________________________________________________________________ 
Second Attempt: (Visit ___, Date: ___/___/___) 
 
1.  Did skin sensitization occur?     Yes 
 No 
 
SKIN 
REACTION 
SCORE 
0 
1+ 
2+ 
3+ 
4+ 
 
 
 
2. Did the subject experience any adverse reactions to the sensitizing dose? 
     Yes          No   
    * If yes, note reaction and file AE form:   ______________________________________ 
____________________________________________________________________________ 
____________________________________________________________________________ 
 
 

 Page 48 
  
Part II: Dose Determination 
 
(Visit ___, Date: ___/___/___) 
 
1.  Complete the grid below with the DTH Reaction score for each region of the dose application matrix.  
 
Concentration 
SKIN 
REACTION 
SCORE 
.2% 
 
.12% 
 
.08% 
 
.04% 
 
 
Immunotherapeutic Response Skin Reaction Scale 
Score  
Observation 
0 
 
No skin reaction 
1+  
 
Erythema only  
2+ 
 
Erythema and cutaneous induration  
3+ 
 
Erythema, papules and small vesicles  
4+ 
 
Large vesicles, bullae, and severe local reaction besides erythema 
 
 
Determined Dose:  ________% 
 
 
2. Did the subject experience any adverse reactions to the dose determination drug application? 
   
   Yes          No   
    * If yes, note reaction and file AE form:   ______________________________________ 
     ____________________________________________________________________________ 
     ____________________________________________________________________________ 
 
 
3. Was first treatment dose applied?             Yes  No 
 
 
 

 Page 49 
  
APPENDIX J: Study Completion Form 
 
Comments: 
 
Please use this space to summarize any issues not covered in the previous forms or to explain procedural deviations: 
 
 
Did subject complete the study?    
 
 
 
                         Yes    No 
If No, Please complete the following: 
Subject Dropped/Withdrawn at Visit #: _______ 
Date subject dropped/withdrawn : ___/___/___ 
 
Check reason for subject withdrawal or termination: 
 
 Sensitization failure: ______________________________________________________________ 
 Adverse Event: ___________________________________________________________________ 
 Subject non-compliance: ___________________________________________________________ 
 Subject withdrawal of Consent: ______________________________________________________ 
 Subject lost to follow-up: ___________________________________________________________ 
 Investigator or Sponsor request to withdraw subject: ____________________________________ 
 Other: __________________________________________________________________________ 
 
Investigator Signature ___________________________                           Date: 
____/_____/____