A Pilot Study to Compare the Efficacy of Pilocarpine Microneedles With Iontophoresis Method for Sweat Induction in Healthy Human Subjects
Phase: NA
Conditions: Cystic Fibrosis
Sponsor: Emory University

 
 
 
NCT04732195 
A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
Date: 11.01.2021 

Page 1 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
 
 
PROTOCOL TITLE: 
A pilot study to compare the efficacy of Pilocarpine Microneedles with iontophoresis 
method for sweat induction in healthy human subjects 
 
PRINCIPAL INVESTIGATOR: 
Lokesh Guglani, MD FAAP 
Associate Professor, 
Division of Pulmonology, Allergy/Immunology, Cystic Fibrosis and Sleep (PACS) 
Department of Pediatrics 
Emory University School of Medicine 
(404) 712-2324 
lokesh.guglani@Emory.Edu 
CO-INVESTIGATOR: 
Mark Prausnitz PhD 
Regents’ Professor and J. Erskine Love Jr. Chair in Chemical & Biomolecular 
Engineering 
School of Chemical and Biomolecular Engineering 
Georgia Institute of Technology 
311 Ferst Drive 
Atlanta, GA 30332-0100 USA 
Phone: +1 (404) 894-5135 
Fax: +1 (404) 894-2291 
Email: prausnitz@gatech.edu 
 
 
VERSION: 
1.3 
FUNDING SOURCE: 
Center for Cystic Fibrosis and Airway Inflammation Research (CF-AIR) Pilot Grant 

Page 2 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
Table of Contents 
1. 
Study Summary .................................................................................................................. 3 
2. 
Objectives ............................................................................................................................ 4 
3. 
Background ......................................................................................................................... 4 
4. 
Study Design ..................................................................................................................... 12 
5. 
Study Endpoints ................................................................................................................ 13 
6. 
Study Intervention/Investigational Agent ....................................................................... 13 
7. 
Procedures Involved ........................................................................................................ 16 
8. 
Data and Specimen Banking........................................................................................... 17 
9. 
Sharing of Results with Participants ............................................................................... 18 
10. 
Study Timelines ............................................................................................................ 18 
11. 
Inclusion and Exclusion Criteria .................................................................................. 18 
12. 
Local Number of Participants ...................................................................................... 18 
13. 
Recruitment Methods ................................................................................................... 18 
14. 
Withdrawal of Participants ........................................................................................... 19 
15. 
Risks to Participants ..................................................................................................... 19 
16. 
Potential Benefits to Participants ................................................................................ 20 
17. 
Compensation to Participants ..................................................................................... 20 
18. 
Data Management and Confidentiality ....................................................................... 20 
19. 
Economic Burden to Participants ............................................................................... 20 
20. 
Consent Process .......................................................................................................... 21 
21. 
Setting ............................................................................................................................ 21 
22. 
Pilocarpine Microneedle Patch Composition ............................................................ 21 
23. 
Pilocarpine Microneedle Patch Manufacturing and Characterization .................... 21 
24. 
Pilocarpine Microneedle Patch Measurements and Disposal after use ................ 22 
25. 
Appendices .................................................................................................................... 22 
26. 
References .................................................................................................................... 23 

Page 3 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
                 Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
1. Study Summary 
 
Study Title 
A pilot study to compare the efficacy of Pilocarpine 
Microneedles with iontophoresis method for sweat induction 
in healthy human subjects
Study Design 
Phase 1 Clinical Pilot study – cross sectional 
Primary Objective 
To compare the sweat volume from application of 
Pilocarpine Microneedle patch to the forearm of healthy 
adult subjects with that of standard of care method using 
pilocarpine iontophoresis
Secondary Objective(s) 
To measure the sweat chloride concentrations from the 
sweat samples obtained from the application of Pilocarpine 
Microneedle patch to the forearm of healthy adult subjects 
and compare them with measurements from standard of 
care method using pilocarpine iontophoresis 
Research 
Intervention(s)/Interactions 
To perform sweat testing on forearms with standard 
pilocarpine iontophoresis method and microneedle patch 
method
Study Population 
Healthy adults >18 years of age 
Sample Size 
50 subjects 
Study Duration for 
individual participants 
1 day 
Study Specific 
Abbreviations/ Definitions 
AE – Adverse Event/Adverse Experience 
CF – Cystic Fibrosis 
CFR – Code of Federal Regulations 
CHOA – Children’s Healthcare of Atlanta 
CTACE – Common Terminology Criteria for Adverse Events 
FDA – Food and Drug Administration 
GCP – Good Clinical Practice 
HIPAA – Health Insurance Portability and Accountability Act 
IB – Investigator’s Brochure 
ICF – Informed Consent Form 
IRB – Institutional Review Board 
N – Number (typically refers to subjects) 
PHI – Protected Health Information 
PI – Principal Investigator 
REDCap – Research Electronic Data Capture 
SAE – Serious Adverse Event 
SOP – Standard Operating Procedure 
Funding Source (if any) 
Center for Cystic Fibrosis and Airway Inflammation 
Research (CF-AIR)

Page 4 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
 
2. Objectives 
 
The main objective of this study is to demonstrate the safety and feasibility of using 
pilocarpine microneedles for sweat induction during the process of sweat testing in 
healthy human adult subjects (>18 years of age) and comparing it to the standard 
method of sweat induction (Pilocarpine iontophoresis) in terms of sweat volume 
collected and sweat chloride concentrations. 
3. Background 
 
The sweat test remains the gold standard test for the diagnosis of Cystic Fibrosis (CF). 
This test is performed in clinical labs and hospitals around the world by clinical personnel 
for collecting sweat from the forearms to measure the sweat chloride concentration. The 
current technique for sweat testing has remained unchanged since it was first 
standardized in the 1960s. Performing this test in newborn infants who have a positive 
initial newborn screen for CF is clinically very important, as early diagnosis and treatment 
has helped to improve long-term growth and respiratory outcomes1. However, only 
accredited CF centers generally have the ability to carry out sweat tests. In addition, a 
common clinical problem encountered in all CF centers is the collection of inadequate 
volumes of sweat during the testing of young children, which necessitates repeat testing. 
This can lead to delays in diagnosis and treatment, and also causes significant anxiety2 
for the parents of the newborn who are anxiously waiting to find out if their child has a 
chronic life-limiting condition3. 
This failure in collecting adequate sweat is especially common in infants less than 3 
months of age, and has been shown to be related to several factors such as gestational 
age4, ethnicity5 and skin factors6. While newborns are capable of producing sweat, the 
failure of sweat collection is mostly due to inadequate stimulation of sweat production by 
iontophoretic delivery of pilocarpine using electricity. Pilocarpine is a cholinergic agonist 
medication that has been in clinical use since 1874 and it stimulates the sweat glands to 
produce more sweat. The current sweat testing method (Figure 1) uses a gel disc 
containing Pilocarpine that drives the medication into the skin with a small electric current 
(iontophoresis) followed a 30-minute period of sweat collection. Given that it is such an 
important clinical test, there is an unmet medical need to improve the current testing 
method and develop more reliable alternatives to induce sweating to ensure adequate 
sample collection that all CF centers are able to carry out. 
 
In collaboration with Dr. Prausnitz from Georgia Tech, we have developed microneedle 
(MN) patches to administer pilocarpine to induce sweating in a simple, reliable way 
without electrical current that does not need special training and does not require 
sophisticated iontophoretic equipment. As shown in Figure 2, each MN patch contains an 
array of solid, water-soluble, micron-scale needles that encapsulate the medication7. 
Upon application to skin, the MNs penetrate into the skin’s upper layers and dissolve in 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
the interstitial fluid to release the loaded drugs. MN patches are painless and can be 
administered with little or no training8. 
 
MN patches have been extensively studied at Georgia Tech and elsewhere, mostly for 
vaccination and systemic drug delivery, including successful clinical trials9. Pilocarpine 
delivery has been studied using metal microneedles coated with pilocarpine for 
administration to the eye for its cholinergic effects10 and as a drug-free pretreatment of 
the skin to create micropores to increase efficiency of pilocarpine delivery by conventional 
iontophoresis11. We have developed MN patches that deliver pilocarpine to skin to 
simplify reliable sweat induction for CF sweat testing without the use of electrical current. 
Therefore, the impact of our work is to translate a cutting-edge technology (MN patches) 
to address an unmet medical need (reliable sweat testing) to increase test reliability, 
enable faster medical treatment, decrease costs and reduce parental anxiety. 
 
 
Figure 1: Current Standard of Care Sweat Testing Procedure. (A) Pilocarpine gel discs 
(arrows) placed on positive and negative electrodes. (B) Pilocarpine iontophoresis using 
1.5mA current to drive pilocarpine into the skin at the red electrode. (C) Collection of 
sweat from the stimulated area of the skin on the forearm in a newborn. 
 
 
 
 
Figure 2. Microneedles patch. (A) A patch with a square array of microneedles in 
the center. (B) a microneedle patch being applied to the skin. (C) A magnified 
view showing a patch with an array of 100 microneedles. 
 
Because the pilocarpine microneedle patch is a drug-device combination, it is important 
to understand the salient characteristics of pilocarpine. The next section provides a 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
detailed overview of pilocarpine – its pharmacologic properties, doses, indications for 
use and adverse effects that have been reported in humans. 
 
Pilocarpine 
 
Figure 3: Chemical structure of Pilocarpine 
Source of Pilocarpine 
 
Pilocarpine is an alkaloid derived from the plant belonging to genus Pilocarpus 
microphyllus that grows in South America. Although the natives of Brazil had long 
known that the chewing of leaves of Pilocarpus plants caused salivation, the active 
compound, pilocarpine, was isolated only in 1875 and shown to affect the pupil and 
sweat and salivary glands. 
Pilocarpine is extracted from the powdered leaf material in a multi-step process. First 
the material is treated with ethanol acidified with hydrochloric acid, and the solvents 
removed under reduced pressure. The resultant aqueous residue is neutralized with 
ammonia and put aside until the resin has completely settled. It is then filtered and 
concentrated by sugar solution to a small volume, made alkaline with ammonia, and 
finally extracted with chloroform. The solvent is removed under reduced pressure to 
create pilocarpine powder which can be used to make pills or eye drops. 
 
Physicochemical Properties of Pilocarpine 
Pilocarpine is soluble in water, alcohol and chloroform. It is sparingly soluble in ether, 
benzene. Pilocarpine is hygroscopic in nature and sensitive to light. It is commercially 
available in powder and liquid suspension form. 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
Molecular Weight: 271.273 g/mol 
Melting Point: 173.5-174 degree Celsius 
Log P: 1.1 
pKa: 1.6, 1.7 
Mechanism of action 
 
Pilocarpine has a dominant muscarinic action but is a partial rather than full agonist; the 
sweat glands are particularly sensitive to pilocarpine. 
Systemically, pilocarpine increases the effect of parasympathetic nervous system and 
decreases the heart rate. As a muscarinic receptor agonist, pilocarpine produces 
vascular relaxation and conjunctival vasodilation. 
In the eye, Pilocarpine directly stimulates cholinergic receptors causing miosis (by 
contraction of the iris sphincter), loss of accommodation (by constriction of ciliary 
muscle), and lowering of intraocular pressure (with decreased resistance to aqueous 
humor outflow). Therefore, it used for reduction of elevated intraocular pressure (IOP) in 
open-angle glaucoma or ocular hypertension; management of acute angle-closure 
glaucoma; induction of miosis (FDA approved in pediatric patients and adults) and 
prevention of postoperative elevated IOP associated with laser surgery (FDA approved 
in adults) 
When taken in the pill form, it binds to muscarinic (cholinergic) receptors, causing an 
increase in secretion of exocrine glands (such as salivary and sweat glands) and 
increase tone of smooth muscle in gastrointestinal and urinary tracts. Its main clinical 
use in the pill form is for the treatment of xerostomia (dry mouth) caused by radiation 
therapy in patients with head and neck cancer and from Sjögren's syndrome. This is 
because Pilocarpine stimulates the salivary gland and stimulates the salivary flow. 
 
When applied to the skin, Pilocarpine stimulates the muscarinic receptor (M1) of 
myoepithelial cells that surrounds the secretory coils of sweat gland and cause 
contraction of myoepithelial cells and facilitates secretion of sweat. 
Pharmacokinetics 
Onset of action: 20 minutes; Maximum effect: 1 hour 
Duration: 3 to 5 hours 
Half-life elimination: 0.76 to 1.35 hours; mild to moderate hepatic impairment: 2.1 hours 
Time to peak, serum: 0.85 to 1.25 hours (increased to 1.47 hours with a high-fat meal) 
Excretion: Urine 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
In the eye, 
Onset of action: Miosis: 10 to 30 minutes; Intraocular pressure reduction: 1 hour. 
Duration: Miosis: 4 to 8 hours; Intraocular pressure reduction: 4 to 14 hours. 
Pharmacodynamics 
Pilocarpine is metabolized by the liver and its dose needs to be adjusted in individuals 
with liver disease. It is eliminated by the kidneys and its excretion can be accelerated by 
acidification of the urine to trap the cationic form in the urine. The characteristics of oral 
pilocarpine are fast onset, short duration of action and variable therapeutic dose 
required by patients. More than half of the oral dose is excreted in urine as unchanged 
drug. 
 
Clinically used doses of Pilocarpine (Source: Lexicomp) 
A. Pill Form 
Xerostomia: Oral: 
Associated with head and neck cancer: Initial: 5 mg 3 times daily; may 
titrate dose based on response and tolerability; usual dosage range: 15 to 30 
mg/day; maximum: 10 mg/dose 
Sjögren syndrome: 5 mg 4 times daily 
B. Eye Drops 
Dosing: Pediatric 
Glaucoma, open-angle; elevated intraocular pressure: 
Infants and Children <2 years: Ophthalmic: 1% Solution: Instill 1 drop into 
the affected eye(s) 3 times daily 
Children ≥2 years and Adolescents: Note: Strength of solution and 
frequency of instillation dependent on degree of pressure elevation and patient 
miotic response; individualize therapy. Ophthalmic: 1%, 2%, or 4% solution: Instill 
one drop into the affected eye(s) up to 4 times daily; for pilocarpine-naive 
patients, initiate therapy with the 1% concentration 
Glaucoma, acute angle closure: 
Infants and Children <2 years: Ophthalmic: 1% solution: Instill 1 drop into 
the affected eye(s) 3 times daily 
Children ≥2 years and Adolescents: Ophthalmic: 1% or 2% solution: Initial: 
Instill 1 drop into the affected eye(s) up to 3 times over a 30-minute period; 
pretreatment with secretory suppressant and hyperosmotic agent may be 
required to lower IOP below 50 mm Hg and relieve iris ischemia. If laser 
iridoplasty or iridomy required to break the attack, instill 1 drop of 4% solution 

Page 9 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
prior to the procedure; following laser iridoplasty, instill 1 drop of 1% solution 4 
times daily until an iridotomy can be performed. 
Miosis induction; prior to goiniotomy or trabeculectomy: 
Infants and Children: Ophthalmic: 1% or 2% solution: Instill 1 drop into the 
eye(s) 15 to 60 minutes prior to surgery 
Adolescents: Ophthalmic: 1%, 2%, or 4% solution: Instill 1 or 2 drops (5 
minutes apart) into the affected eye(s) 
Dosing: Adult 
Elevated intraocular pressure: Ophthalmic: 
Open-angle glaucoma or ocular hypertension: Instill 1 drop of 1%, 2%, or 
4% solution into the affected eye(s) up to 4 times daily; initiate pilocarpine-naive 
patients on the 1% concentration. Note: Strength of solution and frequency of 
instillation dependent on degree of pressure elevation and patient miotic 
response. 
Angle-closure glaucoma, acute: Instill 1 drop of 2% solution into the 
affected eye as part of a 4-drug regimen; may repeat in 30 to 60 minutes if 
intraocular pressure remains elevated (eg, >40 mm Hg). Note: Reserve medical 
management for emergency situations when an assessment by an 
ophthalmologist will be delayed by ≥1 hour (Pokhrel 2007). 
Miosis: Ophthalmic: Instill 1 drop (or 2 drops 5 minutes apart) of 1%, 2%, 
or 4% solution into the affected eye(s). 
Prevention of postoperative elevated intraocular pressure: 
Ophthalmic: Instill 1 drop (or 2 drops 5 minutes apart) of 1%, 2%, or 4% solution 
into the affected eye(s) 15 to 60 minutes prior to surgery. 
 
Commercially Available Pilocarpine Brand Names and Products 
Eye Drops: 
Isopto Carpine: 1% (15 mL); 2% (15 mL); 4% (15 mL) 
Generic: 1% (15 mL); 2% (15 mL); 4% (15 mL) 
Pills:  
 
Salagen: 5 mg 
Salagen: 7.5 mg [contains fd&c blue #2 aluminum lake] 
Generic: 5 mg, 7.5 mg 
 
Side effects of Pilocarpine therapy 
Eye Drops: 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
• Tearing 
• Burning 
• Blurred vision 
• Decreased night vision 
 
Systemic: 
The following adverse drug reactions and incidences are derived from product labeling 
unless otherwise specified. 
>10%: 
Cardiovascular: Flushing (8% to 13%) 
Central nervous system: Chills (3% to 15%), dizziness (5% to 12%), headache (11%) 
Gastrointestinal: Nausea (6% to 15%) 
Genitourinary: Urinary frequency (9% to 12%) 
Neuromuscular & skeletal: Weakness (2% to 12%) 
Respiratory: Rhinitis (5% to 14%) 
Miscellaneous: Diaphoresis (29% to 68%) 
1% to 10%: 
Cardiovascular: Edema (<1% to 5%), facial edema, hypertension (3%), palpitation, 
tachycardia 
Central nervous system: Pain (4%), fever, somnolence 
Dermatologic: Pruritus, rash 
Gastrointestinal: Diarrhea (4% to 7%), dyspepsia (7%), vomiting (3% to 4%), 
constipation, flatulence, glossitis, salivation increased, stomatitis, taste perversion 
Genitourinary: Vaginitis, urinary incontinence 
Neuromuscular & skeletal: Myalgias, tremor 
Ocular: Lacrimation (6%), amblyopia (4%), abnormal vision, blurred vision, conjunctivitis 
Otic: Tinnitus 
Respiratory: Cough increased, dysphagia, epistaxis, sinusitis 
Miscellaneous: Allergic reaction, voice alteration 
<1%: Abnormal dreams, abnormal thinking, alopecia, angina pectoris, anorexia, anxiety, 
aphasia, appetite increased, arrhythmia, arthralgia, arthritis, bilirubinemia, body odor, 
bone disorder, bradycardia, breast pain, bronchitis, cataract, cholelithiasis, colitis, 
confusion, contact dermatitis, cyst, deafness, depression, dry eyes, dry mouth, dry skin, 
dyspnea, dysuria, ear pain, ECG abnormality, eczema, emotional lability, eructation, 
erythema nodosum, esophagitis, exfoliative dermatitis, eye hemorrhage, eye pain, 
gastritis, gastroenteritis, gastrointestinal disorder, gingivitis, glaucoma, hematuria, 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
hepatitis, herpes simplex, hiccup, hyperkinesias, hypoesthesia, hypoglycemia, 
hypotension, hypothermia, insomnia, intracranial hemorrhage, laryngismus, laryngitis, 
leg cramps, leukopenia, liver function test abnormal, lymphadenopathy, mastitis, 
melena, menorrhagia, metrorrhagia, migraine, moniliasis, myasthenia, MI, neck pain, 
photosensitivity reaction, nervousness, ovarian disorder, pancreatitis, paresthesia, 
parotid gland enlargement, peripheral edema, platelet abnormality, pneumonia, pyuria, 
salivary gland enlargement, salpingitis, seborrhea, skin ulcer, speech disorder, sputum 
increased, stridor, syncope, taste loss, tendon disorder, tenosynovitis, 
thrombocythemia, thrombocytopenia, thrombosis, tongue disorder, twitching, urethral 
pain, urinary impairment, urinary urgency, vaginal hemorrhage, vaginal moniliasis, 
vesiculobullous rash, WBC abnormality, yawning 
Drug Interactions 
Acetylcholinesterase Inhibitors: May enhance the adverse/toxic effect of Cholinergic 
Agonists. Specifically, cholinergic effects may be enhanced or increased. Risk C: 
Monitor therapy 
Beta-Blockers: May enhance the adverse/toxic effect of Cholinergic Agonists. Of 
particular concern are the potential for cardiac conduction abnormalities and 
bronchoconstriction. Risk C: Monitor therapy 
Cimetropium: Cholinergic Agonists may diminish the anticholinergic effect of 
Cimetropium. Risk C: Monitor therapy 
Sincalide: Drugs that Affect Gallbladder Function may diminish the therapeutic effect of 
Sincalide. Management: Consider discontinuing drugs that may affect gallbladder 
motility prior to the use of sincalide to stimulate gallbladder contraction. Risk D: 
Consider therapy modification 
Food Interactions 
Fat decreases the rate of absorption, maximum concentration and increases the time it 
takes to reach maximum concentration. Management: Avoid administering with a high- 
fat meal. 
Pregnancy Considerations 
Adverse events have been observed in some animal reproduction studies. 
Breast Feeding Considerations 
It is not known if pilocarpine is excreted in breast milk. Due to the potential for serious 
adverse reactions in the nursing infant, the manufacturer recommends a decision be 
made to discontinue nursing or to discontinue the drug, taking into account the 
importance of treatment to the mother. 
FDA Approval Date 
Pilocarpine Hydrochloride: March 22, 1994 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
4. Study Design 
 
This is a cross sectional pilot study which will be conducted in a phased manner (Figure 
4). The first phase of the study will include the enrollment of 10 healthy adult subjects 
who will undergo testing to assess the safety and efficacy of pilocarpine microneedle 
patch. The safety data from the first 10 subjects will be submitted to the Data and Safety 
Monitoring Board (DSMB) that will review the reported adverse events during testing. 
The DSMB will independently assess this safety data and provide their 
recommendations on whether it is safe to continue the study or not. These 
recommendations will be submitted to the Emory IRB. If there are no safety concerns 
reported by DSMB, then the second phase of the study will begin where the study team 
will continue enrollment of an additional 40 subjects to complete testing. After the 
enrollment and testing of 50 subjects, the study will be completed and another report of 
safety data will be submitted to the DSMB at the end of the study. The DSMB will review 
this final report and their final recommendations will be shared with Emory IRB and 
study team. 
Figure 4: Study Design Flowchart 

Page 13 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
If there are any safety concerns reported by the DSMB, then these will be addressed by 
the study team with an appropriate modification of the study design or intervention. This 
will be followed by submission of an amended study protocol to the IRB to reflect the 
changes made to the study protocol. After approval of the amended study protocol, the 
study team will conduct the revised testing procedure on 10 subjects and submit safety 
data to the DSMB again for their review and approval. 
5. Study Endpoints 
 
The study endpoint will be when the primary and secondary outcome measures are 
obtained for all the enrolled participants and data analysis has been completed. 
 
The main outcome measures for the study are: 
5.1 To compare the sweat volume from application of Pilocarpine Microneedle 
patch to the forearm of healthy adult subjects with that of standard of care 
method using pilocarpine iontophoresis 
5.2 To measure the sweat chloride concentrations from the sweat samples 
obtained from the application of Pilocarpine Microneedle patch to the 
forearm of healthy adult subjects and compare them with measurements 
from standard of care method using pilocarpine iontophoresis 
 
6. Study Intervention/Investigational Agent 
 
The study involves the use of pilocarpine for its approved indication for sweat induction 
during the clinical procedure for sweat testing in human subjects. The pilocarpine will be 
administered in the form of an agar gel disc using iontophoresis (Standard Procedure) 
on one arm of the subject, while the other arm will receive pilocarpine microneedles 
(Experimental Procedure) without any iontophoresis. This will allow a direct comparison 
of the two methods in terms of the efficiency of induction of sweat production in human 
subjects and whether the sweat chloride concentrations in the sweat samples obtained 
by the two methods are comparable or not. In addition, a control microneedle patch 
without any Pilocarpine will be placed on the left forearm to assess the effect of the 
microneedle patch use. The subsequent steps for the collection of sweat after 
completing the induction will be done with the same method for both arms using a 
macroduct collector at each site (Standard Procedure). There will be no sweat collection 
performed at the site the control microneedle patch. Pictures of the skin sites will be 
taken before and after the interventions at each site. 
 
The study team will plan to enroll study subjects of all ethnicities or racial backgrounds, 
so that the impact of this intervention can be studied in individuals with different skin 
characteristics. 

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IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
Because this study involves an approved drug (Pilocarpine) that has been in clinical use 
for application to the skin for the purpose of sweat testing, we think that this project will 
not require an Investigational New Drug (IND) application. This has been discussed at 
length with several regulatory experts and FDA and the conclusion of these discussions 
was that the drug component is exempt from an IND under 21 CFR 312.2(b), and the 
device component is exempt from an IDE under 21 CFR 812.2(b), since all the related 
conditions in those two provisions are met by our device and study design. 
 
We have reviewed the guidelines from the Food and Drug Administration (FDA)12 for 
determining whether certain human research studies can be conducted without an IND 
(From FDA website: https://www.federalregister.gov/documents/2015/10/30/2015- 
27729/investigational-new-drug-applications-determining-whether-human-research-   
studies-can-be-conducted ). 
 
This document provides the criteria that FDA uses to determine if any human subjects 
research study involving a drug needs an IND or not. We have extracted the 6 main 
criteria from this FDA advisory document and provided an explanation of how our study 
meets those criteria set forth by the FDA. 
 
1) The drug product is lawfully marketed in the United States. 
 
For our proposed study, we are planning to use Pilocarpine, which is available for use in 
United States and is approved by the FDA for standardized induction of sweat 
production during sweat testing. The Sweat Test is the gold standard clinical test for the 
diagnosis of Cystic Fibrosis, the most common lethal genetic disease in Caucasians. 
This test involves collection of a small amount of sweat from the forearms after the 
application of an agar gel disc containing pilocarpine with iontophoresis using a small 
electric current (1.5mA) for 5 minutes. 
 
Pilocarpine is a cholinergic agonist agent with the ability to induce sweat production 
from eccrine human sweat glands based on its ability to stimulate muscarinic receptors 
in the sweat gland ducts. After the stimulation phase with pilocarpine, the sweat 
produced by the skin on the forearm is collected and analyzed for its chloride 
concentration. Improving upon the current technique of sweat testing will reduce 
inaccurate or inadequate results and improved diagnostic certainty in newborn babies 
that are screened for Cystic Fibrosis at birth across United States and most of Europe 
as well. 
 
2) The investigation is not intended to be reported to FDA as a well-controlled study 
in support of a new indication and there is no intent to use it to support any other 
significant change in the labeling of the drug. 
This study will not be reported to the FDA as a case-control study as we are planning to 
study the feasibility of pilocarpine delivery to the skin using microneedle patches in 

Page 15 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
healthy subjects only. The results of this study will compare the effectiveness of the 
microneedle method of sweat induction with the standard of care pilocarpine 
iontophoresis method in human subjects and will not change the labeling of the drug. 
The sweat collection phase and analysis of chloride concentration will remain in the 
same and we will simply compare the volume of sweat collected between the standard 
method of sweat induction (using iontophoresis) with our proposed microneedle-based 
approach. 
3) In the case of a prescription drug, the investigation is not intended to support a 
significant change in the advertising for the drug. 
 
This study and the data being collected from it will not be used in any advertising 
materials for the drug. The same drug (pilocarpine) is being used for sweat stimulation 
in this study and we are simply comparing the effectiveness of another mode of delivery 
of this medication. If found effective, it will not change the use of this drug for the 
purpose of sweat testing. 
 
4) The investigation does not involve a route of administration, dose, patient 
population, or other factor that significantly increases the risk (or decreases the 
acceptability of the risk) associated with the use of the drug product (21 CFR 
312.2(b)(1)(iii)). 
 
The delivery of pilocarpine for sweat induction using the microneedle approach does not 
change the route of administration (transcutaneous) for the purpose of sweat testing. 
The dose used in microneedles is significantly lower and does not increase the risk for 
the subjects being tested. The patient population that this test is applicable to remains 
the same – all subjects that are being evaluated for Cystic Fibrosis. There is no change 
in the risk profile, as the drug is being applied to the skin for the stimulation of the sweat 
glands on the forearm. Since our proposed microneedle method does not involve the 
use of electric current, there will actually be a lower risk of tingling sensations and rarely 
skin burns that sometimes occur with the iontophoresis procedure. The standard 
pilocarpine iontophoresis method of sweat induction has been rarely reported to cause 
small burns on the skin surface related to the use of electric current. Our proposed 
method eliminates this risk of burns completely. 
In particular, the FDA advisory document (page 5)12 provides the following explanation 
regarding the risk of the modifications made in the proposed study: 
 
In responding to comments asking FDA to clarify to what extent a sponsor could 
change the marketed drug product or conditions of use and still be exempt from 
the IND regulations, FDA stated that: 
The exemption was not intended to require an investigator to use the drug in 
exactly the same dosage form, dosage levels, and patient populations described 
in the marketed labeling for the product, but rather to permit changes to the 

Page 16 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
lawfully marketed drug product that do not increase the risks . . . over the risk 
presented by use of the product in conformance with its marketed labeling 
 
Since our proposed study using microneedle-based delivery of pilocarpine reduces the 
risk associated with standard of care iontophoresis method, it satisfies this criterion as 
well. 
 
5) The investigation is conducted in compliance with the requirements for review by 
an IRB (21 CFR part 56) and with the requirements for informed consent (21 
CFR part 50). 
 
The study will be submitted for evaluation by Emory Institutional Review Board and all 
the stated rules and regulations will be followed by the principal investigator during the 
conduct of the study. An informed consent will be signed by every participant prior to 
undergoing any testing and the subjects will be informed about the voluntary nature of 
the study and that they have the right to withdraw consent for this study at any given 
time. Their data will be anonymized and stored in a HIPAA compliant manner in a 
password protected secure network computer. 
6) The investigation is conducted in compliance with the requirements of § 312.7 
(i.e., the investigation is not intended to promote or commercialize the drug 
product). 
 
The study will be conducted by researchers from Emory University and Georgia Institute 
of Technology without any intention of promoting any commercial drug products. 
 
Drug/Device Handling: The supplies for the Standard Procedure will be obtained from 
the manufacturer directly and stored according to their specifications. The pilocarpine 
microneedles that will be used for the experimental procedure will be manufactured at 
Global Center for Medical Innovation (GCMI). This is not part of Georgia Tech. It is an 
independent institution. The patches will be delivered to Emory University by a member 
of Dr. Prausnitz lab. The patches will then be stored at room temperature at Emory 
University in a secure location until it is used for a study subject. 
 
 
7. Procedures Involved 
 
Informed consent will be obtained by an IRB-approved member of the study staff. The 
process of informed consent will begin with a verbal description of the experimental 
tasks. Participants will then have an opportunity to read the informed consent form that 
explains in more detail the requirements for participation, including protections of 
privacy and confidentiality. Participants will be told of their rights to discontinue 
participation at any time and that there will be no immediate benefit to participating and 
no disadvantage to discontinuing. Participants will be given contact numbers for the PI 
and the Executive Director of the Human Research Protection Office should they have 

Page 17 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
any questions or concerns regarding their participation. Informed consent 
documentation will take place in a private setting convenient to the participant. 
 
Once the consent is obtained, the patient will undergo sweat testing. The testing will be 
performed with the subject resting comfortably on a chair and total duration of testing 
will be 45 minutes. The right forearm will be used for pilocarpine iontophoresis and the 
left forearm will be used for microneedle-based stimulation for a period of 5 minutes. A 
control microneedle patch without pilocarpine will also be placed on the left forearm to 
assess for any microneedle patch related changes. After the stimulation phase, 
collection of sweat will be performed on each forearm simultaneously for a period of 30 
minutes (excluding the control microneedle patch site). At the end of the 30-minute 
sweat collection period, the collection device will be removed and the study visit will be 
completed. All the collected sweat samples (2 for each subject) will be frozen for later 
analysis of the sweat volume and chloride concentration. 
The pilocarpine microneedles that will be used for the experimental procedure will be 
manufactured at Global Center for Medical Innovation (GCMI). This is not part of Georgia 
Tech. It is an independent institution. The pilocarpine-loaded MN patches are fabricated 
by a two-step molding process using polydimethylsiloxane (PDMS) molds with some 
modification on an established method. The first casting solution is a mixture of 10% (w/v) 
pilocarpine nitrate, 10% (w/v) poly (vinyl alcohol) and 5% (w/v) sucrose, which is prepared 
by mixing the different components in water at the desired ratio. This solution is then cast 
on PDMS molds under vacuum to facilitate filling the solution into the molds. After 20 min, 
excess solution is removed, and the filled molds are centrifuged at 5000 rpm for 20 min 
to dry the drug loaded MNs. The second casting solution containing 20% (w/w) 
polystyrene in 1, 4-dioxane is then cast on the filled PDMS molds under vacuum. The 
mold is kept under vacuum for another 3 h to dry the solution at room temperature, and 
then further dried at 40°C overnight before demolding the MN patches by adhesive tape. 
Each patch is immediately stored with desiccant in individual sealed pouches prior until 
use. 
8. Data and Specimen Banking 
Data for the study participants will be stored in the online REDCap (Research 
Electronic Data Capture) Database with only de-identified information being used 
for statistical analyses. REDCap is a secure, web-based application designed to 
support data capture for research studies, providing 1) an intuitive interface for 
validated data entry; 2) audit trails for tracking data manipulation and export 
procedures; 3) automated export procedures for seamless data downloads to 
common statistical packages; and 4) procedures for importing data from external 
sources. Access to this data will be limited to the study investigators only. Sweat 
samples may be stored frozen at Emory Children’s Center for future tests. All the 
used microneedle patches will be sent to Prausnitz lab to measure the amount of 
Pilocarpine delivered into the skin and then the patches will be destroyed as per 
the steps outlined in Appendix 8. 

Page 18 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
9. Sharing of Results with Participants 
The study team does not plan to share the results of the sweat sample collection. 
Sweat samples will be tested for chloride concentration and volume. The 
summary results will be posted on clinicaltrials.gov website. 
 
10. Study Timelines 
Participants will only need to take part in one study visit totaling 45 minutes. The 
study team anticipates enrolling all study participants over the course of 6 
months. Completion of all study related procedures, including data analysis, is 
expected to take approximately 12 months. No follow up visits will be needed for 
any of the study participants. 
 
11. Inclusion and Exclusion Criteria 
Inclusion Criteria: 
 Age >18 years 
 Signed a written informed consent 
 Not taking any medications 
 No known medical diagnoses or chronic conditions 
 
Exclusion Criteria: 
 Age <18 years 
 Family history of Cystic Fibrosis 
 History of skin disorders (eczema, psoriasis etc.) that could prevent 
sweat testing on forearms 
 Current medication use 
 Pregnant women 
 Breastfeeding women 
12. Local Number of Participants 
We will enroll 50 healthy adults aged 18 years or more from the general 
community with the help of flyers that will be placed at various places at the 
Emory campus. There will be no exclusion of subjects based on gender or 
ethnicity. 
 
 
13. Recruitment Methods 
 
Participants will be recruited through flyers and consent will be obtained during their 
scheduled study visit. Recruited subjects will not require any follow up visits or 
additional testing. 

Page 19 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
14. Withdrawal of Participants 
Subjects will only be withdrawn from the study if they are unable to complete the 
sweat sample collection. After a subject has been withdrawn, the study team will 
discard incomplete sweat sample collection and no more data will be collected. 
 
15. Risks to Participants 
There are some risks associated with both forms of sweat testing. 
All study subjects will be warned about the potential risk of skin burns that can occur 
with the use of the pilocarpine iontophoresis step during the standard-of-care method of 
sweat testing. 
The product insert from the manufacturer detailing this side effect has been added as 
Appendix 10. This risk is estimated to occur at a rate of about 1 in 50,000 tests. The 
burns have been reported to be ranging from “tiny black pinholes in the skin” to “crater- 
like, third degree burns two to three millimeters in diameter”. As per the manufacturer’s 
product insert, individuals who had these burns had no signs of pain or discomfort 
during iontophoresis, and the burns are not noticed until after the electrodes are 
removed. The burns are reported to heal completely in 1-2 weeks with little or no 
scarring. The manufacturer of pilocarpine iontophoresis also warns about the risk of 
redness of skin at the electrode site, and in some cases, blister-like welts may form 
which can persist up to 2-3 hours. However, this standard-of-care method has been 
approved for use in humans by the FDA. 
 
With the use of pilocarpine microneedle patch, there is a risk of skin irritation or 
redness that can last for hours after the completion of testing. There is no risk of 
burns with the pilocarpine microneedle patch. There is no risk of systemic side 
effects from the exposure to Pilocarpine as the total dose of pilocarpine in each 
microneedle patch is less than 10% (one-tenth) of the standard adult dose that is 
used in clinical practice (see section on Pilocarpine doses above on page 8). Not 
all the pilocarpine that is present on a microneedle patch will be delivered into the 
skin, and only a fraction of that small dose penetrating the skin may be absorbed 
into systemic circulation. 
 
Due to the pilot nature of this study, the study will have a Data and Safety 
Monitoring Board (DSMB) comprised of 3 members that are not affiliated with this 
study who will review the safety data for the first 10 subjects enrolled. The DSMB 
charter has been reviewed and signed off on by all the members of the DSMB 
(Appendix 11). 
 
After the DSMB has reviewed the safety of the pilocarpine microneedle approach in the 
first 10 subjects, it will provide its recommendations to the study team and this 
communication will be submitted to Emory IRB as well. If the DSMB recommends 

Page 20 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
continuation of the study, then further enrollment will be continued to enroll an additional 
40 subjects with close monitoring for any unforeseen risks or side effects of the 
interventions related to this study in the remaining participants. Any unforeseen side 
effects of the pilocarpine microneedle patch will be reported to the DSMB for review. 
The DSMB will again review the safety data at the end of the enrollment of the 
additional 40 subjects to ensure that no additional concerns have arisen during the 
remainder of the study. The DSMB will provide their recommendations and summary 
regarding the safety of the pilocarpine microneedle patch being test in this study and 
this will also be shared with Emory IRB. If there are any safety concerns reported by the 
DSMB, then these will be addressed by the study team with an appropriate modification 
of the study design or intervention. This will be followed by submission of an amended 
study protocol to the IRB to reflect the changes made to the study protocol. After 
approval of the amended study protocol, the study team will conduct the revised testing 
procedure on 10 subjects and submit safety data to the DSMB again for their review 
and approval. 
 
16. Potential Benefits to Participants 
16.1 
There is no direct benefit to the subjects from participation in this 
study. 
16.2 
The study will benefit the advancement of medical knowledge about 
the usefulness of pilocarpine microneedle patches for the induction of 
sweating during the procedure of sweat testing. 
 
 
17. Compensation to Participants 
Subjects will not be compensated for participating in this study. 
18. Data Management and Confidentiality 
Descriptive analysis will be conducted for the study participants to describe their 
clinical presentation and long-term outcomes. Several steps will be taken to 
secure the data by ensuring adequate training of the study team members, 
authorization of access to the study team for the clinical data, password 
protection, encryption, and separation of identifiers and data during the storage, 
use, and transmission. Data will be stored on each users’ secure servers and not 
downloaded to external devices, including laptops. The information, even if de- 
identified, will be destroyed at the expiration of the project. 
 
19. Economic Burden to Participants 
There will be no costs to participants that enroll this study. 

Page 21 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
20. Consent Process 
 
Consent of participants will take place during their scheduled study visit. The research 
coordinator will first introduce the study to the participant and give the participant ample 
time to consider enrolling in the study and to ask questions. The subject will receive a 
copy of their signed consent form. 
 
 
21. Setting 
Study participants will be recruited with the help of study flyers. After participants have 
been screened based on inclusion and exclusion criteria and informed consent 
obtained, their study visit will take place in a designated research room located on the 
first floor of Emory Children’s Center. Study samples that are collected will then be 
taken to the research lab for sample processing. 
22. Pilocarpine Microneedle Patch Composition 
The pilocarpine-loaded MN patches are fabricated by a two-step molding process using 
polydimethylsiloxane (PDMS) molds with some modification on an established method. 
The first casting solution is a mixture of 10% (w/v) pilocarpine nitrate, 10% (w/v) poly (vinyl 
alcohol) and 5% (w/v) sucrose, which is prepared by mixing the different components in 
water at the desired ratio. While this is our current formulation, we will investigate other 
formulations that can increase the pilocarpine content (on a solids basis) and thereby 
increase the dose of pilocarpine per MN. This solution is then cast on PDMS molds under 
vacuum to facilitate filling the solution into the molds. 
After 20 min, excess solution is removed, and the filled molds are centrifuged at 5000 rpm 
for 20 min to dry the drug loaded MNs. The second casting solution containing 20% (w/w) 
polystyrene in 1, 4-dioxane is then cast on the filled PDMS molds under vacuum. The 
mold is kept under vacuum for another 3 h to dry the solution at room temperature, and 
then further dried at 40°C overnight before demolding the MN patches by adhesive tape. 
Each patch is immediately stored with desiccant in individual sealed pouches prior until 
use. 
 
23. Pilocarpine Microneedle Patch Manufacturing and Characterization 
The steps involved in Microneedle patch manufacturing have been outlined in Appendix 
01. 
The device master record showing all the components that went into the research article 
is included in Appendix 2. Design Trace matrix is an engineering description of each 
feature of the microneedle patch and the respective rational for that feature is provided 
in Appendix 3. 

Page 22 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
Verification Testing for the research articles was done through several methods. A 
Mechanical Test Report is included in Appendix 4. Pig skin test report is included in 
Appendix 5. Process validation report is included in Appendix 6. A visual and 
dimensional analysis of the microneedle patch is included in Appendix 7. 
The process of extraction of Pilocarpine Nitrate through HPLC method is described in 
Appendix 8. 
 
A detailed drawing of the research article with all the measurements is included in 
Appendix 9. 
 
24. Pilocarpine Microneedle Patch Measurements and Disposal after use 
The process of determining pilocarpine dose delivered per Microneedle patch will be 
completed at Prausnitz lab. All used patches will be stored in their respective containers 
and transported back to Prausnitz lab. The patches are then dissolved and the 
remaining amount of Pilocarpine in the patch will be measured as per the steps outlined 
in Appendix 8. 
 
25. Appendices 
Appendix 1: Manufacturing Procedure 
Appendix 2: Device Master Record 
Appendix 3: Design Trace Matrix 
Appendix 4: Mechanical Test Report 
Appendix 5: Pig Skin Report 
Appendix 6: Process validation report 
Appendix 7: Visual and Dimensional Measurement Report 
Appendix 8: HPLC Pilocarpine Nitrate extraction Appendix 
9: Detailed Drawing of Research Article 
Appendix 10: Risk of Skin Burns from Pilocarpine Iontophoresis – Manufacturer Product 
Insert 
Appendix 11: Data Safety Monitoring Board Charter 

Page 23 of 23 
IRB Form BIO 08102020 
Protocol Version: 2021.11.01
 
Protocol Title:  A pilot study to compare the efficacy of Pilocarpine Microneedles with 
iontophoresis method for sweat induction in healthy human subjects 
 
 
 
26. References 
1. 
Coffey MJ, Whitaker V, Gentin N, et al. Differences in Outcomes between Early 
and Late Diagnosis of Cystic Fibrosis in the Newborn Screening Era. J Pediatr. 
2017;181:137-145 e131. 
2. 
Moran J, Quirk K, Duff AJ, Brownlee KG. Newborn screening for CF in a regional 
paediatric centre: the psychosocial effects of false-positive IRT results on 
parents. J Cyst Fibros. 2007;6(3):250-254. 
3. 
Hayeems RZ, Miller FA, Barg CJ, et al. Psychosocial Response to Uncertain 
Newborn Screening Results for Cystic Fibrosis. J Pediatr. 2017;184:165-171 
e161. 
4. 
Kleyn M, Korzeniewski S, Grigorescu V, et al. Predictors of insufficient sweat 
production during confirmatory testing for cystic fibrosis. Pediatr Pulmonol. 
2011;46(1):23-30. 
5. 
Eng W, LeGrys VA, Schechter MS, Laughon MM, Barker PM. Sweat-testing in 
preterm and full-term infants less than 6 weeks of age. Pediatr Pulmonol. 
2005;40(1):64-67. 
6. 
LeGrys VA. Sweat testing for cystic fibrosis: profiles of patients with insufficient 
samples. Clin Lab Sci. 1993;6(2):73-74. 
7. 
Arya J, Henry S, Kalluri H, McAllister DV, Pewin WP, Prausnitz MR. Tolerability, 
usability and acceptability of dissolving microneedle patch administration in 
human subjects. Biomaterials. 2017;128:1-7. 
8. 
Norman JJ, Arya JM, McClain MA, Frew PM, Meltzer MI, Prausnitz MR. 
Microneedle patches: usability and acceptability for self-vaccination against 
influenza. Vaccine. 2014;32(16):1856-1862. 
9. 
Rouphael NG, Paine M, Mosley R, et al. The safety, immunogenicity, and 
acceptability of inactivated influenza vaccine delivered by microneedle patch 
(TIV-MNP 2015): a randomised, partly blinded, placebo-controlled, phase 1 trial. 
Lancet. 2017;390(10095):649-658. 
10. 
Jiang J, Gill HS, Ghate D, et al. Coated microneedles for drug delivery to the eye. 
Invest Ophthalmol Vis Sci. 2007;48(9):4038-4043. 
11. 
Wing D, Prausnitz MR, Buono MJ. Skin pretreatment with microneedles prior to 
pilocarpine iontophoresis increases sweat production. Clin Physiol Funct 
Imaging. 2013;33(6):436-440. 
12. 
Services USDoHaH, Administration FaD, (CDER) CfDEaR, (CBER) CfBEaR, 
(CFSAN) CfFSaAN. Investigational New Drug Applications (INDs) — Determining 
Whether Human Research Studies Can Be Conducted Without an IND. In: 
Administration FaD, ed2013