An Open-Label Study of Glutathione in Children With Autism Spectrum Disorder
Phase: PHASE4
Conditions: Autism Spectrum Disorder
Sponsor: University of Chicago

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An Open Label Study of Glutathione in Children with Autism Spectrum Disorder 
Karam Radwan, M.D. 
Kamilah C. Banks-Word, M.D., M.A. 
University of Chicago Medical Center 
Chicago, IL 
 
IRB Code#: 19-0017 
NCT# NCT05954052 
 
Protocol Version 
April 2019 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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An Open Label Study of Glutathione in Children with Autism Spectrum Disorder 
 
This project will take place at the University of Chicago, Department of Child and Adolescent 
Psychiatry. Dr. Karam Radwan, M.D., is the Principal Investigator and can be contacted at 773-
834-6686, email at kradwan@yoda.bsd.uchicago.edu or by mail at 5841 South Maryland Ave (MC 
3077), Chicago, IL 60637. 
 
This clinical trial will be conducted in the spirit of Good Clinical Practice (GCP) and in accordance 
with an IRB approved protocol. No deviation from the protocol will be implemented without the 
prior review and approval of the IRB except where it may be necessary to eliminate an immediate 
hazard to a research subject. In such a case, the deviation will be reported to the IRB as soon as 
possible. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Population  
The population to be studied for this trial are children and adolescents, both male and female, ages 
4-17 that have a current diagnosis of Autism Spectrum Disorder (ASD). 
 
Diagnostic Criteria (1) 
The following diagnostic criteria for ASD will be utilized. Criteria are defined by the Diagnostic 
and Statistical Manual for Mental Disorders (DSM-5) for a diagnosis of Autism Spectrum Disorder 
(1): 
A. Persistent deficits in social communication and social interaction across multiple 
contexts, as manifested by the following, currently or by history (examples are illustrative, not 
exhaustive, see text): 
1.       Deficits in social-emotional reciprocity, ranging, for example, from abnormal social approach 
and failure of normal back-and-forth conversation; to reduced sharing of interests, emotions, or 
affect; to failure to initiate or respond to social interactions. 
2.       Deficits in nonverbal communicative behaviors used for social interaction, ranging, for 
example, from poorly integrated verbal and nonverbal communication; to abnormalities in eye 
contact and body language or deficits in understanding and use of gestures; to a total lack of facial 
expressions and nonverbal communication. 
3.       Deficits in developing, maintaining, and understanding relationships, ranging, for example, 
from difficulties adjusting behavior to suit various social contexts; to difficulties in  sharing 
imaginative play or in making friends; to absence of interest in peers. 
Specify current severity: Severity is based on social communication impairments and restricted 
repetitive patterns of behavior.  
B. Restricted, repetitive patterns of behavior, interests, or activities, as manifested by at 
least two of the following, currently or by history (examples are illustrative, not exhaustive; see 
text): 
1.       Stereotyped or repetitive motor movements, use of objects, or speech (e.g., simple motor 
stereotypies, lining up toys or flipping objects, echolalia, idiosyncratic phrases). 
2.       Insistence on sameness, inflexible adherence to routines, or ritualized patterns or verbal 
nonverbal behavior (e.g., extreme distress at small changes, difficulties with transitions, rigid 
thinking patterns, greeting rituals, need to take same route or eat food every day). 
3.       Highly restricted, fixated interests that are abnormal in intensity or focus (e.g., strong 
attachment to or preoccupation with unusual objects, excessively circumscribed or perseverative 
interest). 
4.       Hyper- or hyporeactivity to sensory input or unusual interests in sensory aspects of the 
environment (e.g., apparent indifference to pain/temperature, adverse response to specific sounds 
or textures, excessive smelling or touching of objects, visual fascination with lights or movement). 
Specify current severity: Severity is based on social communication impairments and restricted, 
repetitive patterns of behavior.  
C. Symptoms must be present in the early developmental period (but may not become fully 
manifest until social demands exceed limited capacities or may be masked by learned strategies in 

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later life). 
D. Symptoms cause clinically significant impairment in social, occupational, or other 
important areas of current functioning. 
E. These disturbances are not better explained by intellectual disability (intellectual 
developmental disorder) or global developmental delay. Intellectual disability and autism 
spectrum disorder frequently co-occur; to make comorbid diagnoses of autism spectrum disorder 
and intellectual disability, social communication should be below that expected for general  
developmental level. 
Note: Individuals with a well-established DSM-IV diagnosis of autistic disorder, 
Asperger’s disorder, or pervasive developmental disorder not otherwise specified should be given 
the diagnosis of autism spectrum disorder. Individuals who have marked deficits in social 
communication, but whose symptoms do not otherwise meet criteria for autism spectrum disorder, 
should be evaluated for social (pragmatic) communication disorder. 
Specify if: 
- With or without accompanying intellectual impairment 
- With or without accompanying language impairment 
- Associated with a known medical or genetic condition or environmental factor 
- Associated with another neurodevelopmental, mental, or behavioral disorder 
- With catatonia  
 
 

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Project Goals  
The goal of the proposed study is to evaluate the safety and feasibility of oral Glutathione in 
children and adolescents who have Autism Spectrum Disorder. Twenty-four subjects with ASD 
will receive 12 weeks of oral Glutathione reduced. The hypothesis to be tested is that: Oral 
Glutathione will be effective in increasing the blood level of Glutathione, which may help to 
decrease some problem behaviors and irritability in this particular ASD population. The second 
aim of this study is to evaluate the tolerability of oral Glutathione. The proposed study may provide 
needed data for future studies aimed at the treatment of aggressive behaviors that can be seen in 
this patient population.  
 
Background and Significance  
Autism Spectrum Disorder is a complex neurodevelopmental disorder, which can involve impaired 
social interaction, impaired language and communication, limited interest, repetitive/ritualistic 
behaviors and sometimes aggressive behavior (3). Although Autism Spectrum Disorder has been 
described in medical literature for years, it remains poorly understood and the exact etiology and 
neurobiology are not quite clear. However, an emerging topic in research is the imbalance between 
bodily oxidative systems and anti-oxidative stress in Autism Spectrum cases (i.e. redox systems). 
Glutathione is involved in neuro-protection against oxidative stress and play a major role in our 
anti-oxidative stress systems, which may mean that decreasing oxidative stress over time could be 
a possible treatment for some problem behaviors that are seen in Autism Spectrum Disorder (3).  
 
Glutathione is a vital antioxidant agent, necessary for neutralizing reactive oxygen species 
(i.e. free radicals) and it is needed for optimal detoxification or removal of toxic substances from 
the human body (5, 7). There is evidence that indicates that children with Autism Spectrum 
Disorder have lower levels of plasma reduced Glutathione than in typically developing children 
(5). This finding has been observed across several neuropsychiatric conditions.  The lower levels 
of glutathione that are seen in this population appear to be due to abnormalities found in the 
transsulfuration pathway, a pathway that is key for Glutathione production (5). The decreased or 
diminished levels of Glutathione can potentially lead to accumulations of reactive oxidative 
species which may cause functional changes in DNA, RNA, protein, lipid and carbohydrate 
systems, leading to cellular dysfunction (4). This redox variance appears to be involved in the 
pathogenesis of ASD and has been proposed by other studies (4).  
According to the Centers for Disease Control, the prevalence of Autism Spectrum Disorder 
in 2014 was about 1 in 59 children (aged 8) in 11 communities across the US and has been 
increasing (2). The disorder occurs in all racial, ethnic and socioeconomic groups; and it is about 
4 times more common among males than females (2). The concern over the last several years has 
been how to effectively treat patients with ASD and especially those with significant social 
impairments and behavioral dysregulation. Some children who have ASD can present with 
irritability, which refers to vocal and motoric outbursts demonstrating anger, frustrations and/or 
distress, often referred to “temper tantrums,” “rages,” or “meltdowns” (8). Along with irritability, 
some children may also present with problem behaviors, which refers to physical acts of aggression 
that have a high potential to result in harm to themselves, other people or property (8). The severity 
of the irritability or problem behaviors can have a significant impact on how the child is perceived 

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by the outside world and can hinder their socialization with others even further. Other very 
important concerns are how the behaviors can jeopardize educational placements and can possibly 
lead to inpatient psychiatric admissions or long-term residential placements (8). 
Although a number of medications have been used to treat comorbidities associated with 
ASD such as neuroleptic agents, antidepressants, stimulants and anxiolytics, there is limited 
evidence about direct pharmacologic treatment of ASD alone. However, complementary and 
alternative medicine has been showing some potential promise in treatment options for this patient 
population. There have been some placebo-controlled trials of N-acetylcysteine (NAC) in ASD 
patients directed toward the treatment of irritability, aggression, self-injurious behavior and 
tantrums (11). Harden et al. (2012), found a significant reduction in irritability symptoms that were 
measured by the Aberrant Behavior Checklist in a placebo-controlled pilot trial of NAC in children 
with ASD (4). Another 12-week randomized, double-blind, placebo-controlled trial of oral NAC 
in children with ASD; found NAC to be safe; however, it was not noted to have a significant impact 
on core social impairment seen in ASD, but the authors did indicate that larger scaled trials are 
needed in order to predict a good treatment response (11). 
N-acetylcysteine is an antioxidant agent and glutamatergic modulator, it is also a precursor 
to Glutathione via cysteine production. Glutathione is found naturally in the human body and 
produced by the liver; it is a 3-amino acid (tripeptide) that is composed of cysteine linked to 
glutamate and followed by glycine attachment (Figure 1). When N-acetylcysteine is de-acetylated, 
cysteine is provided to feed into the transsulfuration pathway, leading to the production of 
Glutathione (9). Glutathione is also found in plants, all mammalian tissue, fungi and some bacteria; 
and it is considered a key determinant of redox signaling, vital in detoxification of xenobiotics, 
regulates cell proliferation, apoptosis and immune function (6, 7). As mentioned, there is some 
evidence that antioxidant agents may be beneficial in reducing irritability or aggression in children 
with ASD. Therefore, we believe that by testing the feasibility of using oral Glutathione directly, 
and examining the anticipated changes in Glutathione blood level, as we are bypassing the 
transsulfuration pathway and providing Glutathione supplementation to the body, may be 
beneficial for ASD patients. Given that Glutathione may have various advantages over other 
pharmacological agents in treating children with ASD, and it is an all-natural supplement, it can 
be a promising treatment for irritability and problem behaviors seen in this population and may 
play a role in correcting  the lower levels of plasma reduced Glutathione often observed in children 
with ASD.  
 
 
 
 
 
 
Figure 1: Image NAC de-acetylation and formation of Glutathione (9)  

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Aims  
1) To determine whether Glutathione given over a 12-week period increases plasma 
Glutathione level in a sample of 24 children aged 4-17 years old with a history of Autism 
Spectrum Disorder. 
2) To determine the feasibility of using oral Glutathione in this patient population 
3) To test whether oral administration of Glutathione supplement is well tolerated as a medical 
food in children with ASD.  
 
Overall, we will examine the effects of Glutathione in children with Autism Spectrum Disorder. 
Evidence suggests that Glutathione may reduce reactive oxygen species that can accumulate in 
patients with ASD and thereby may help to decrease significant problem behaviors seen in this 
population. We hypothesize that oral Glutathione will be safe and well tolerated in patients with 
Autism Spectrum Disorder. 
 
METHODS 
The study will consist of 12 weeks of open label trial with treatment of oral Glutathione in 24 
subjects with ASD; the subjects will be recruited at the University of Chicago. Subjects will be 
seen every two weeks for the first month and then every four weeks for the remaining two months 
with one final visit at the end of the study for a total of 5 visits.  
 
 
 
 
 
Study Location  
This is a single-site trial in the United States, with a total of 24 subjects aged 4-17. The Principal 

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Investigator for this study is: 
 
Karam Radwan, M.D.  
Professor of Child and Adolescent Psychiatry  
University of Chicago  
5841 South Maryland Avenue, Chicago, IL, 60637 
 
All subjects will be seen in the Department of Child and Adolescent Psychiatry at the University 
of Chicago Medical Center at 5841 South Maryland Avenue. The offices are connected to the 
Department of Psychiatry and provide confidential interview rooms and facilities.  
Investigational Agent  
Glutathione (GSH) Reduced (Oral Capsules - Jarrow Brand)  
– for patients 40kg or less: 500 mg by mouth twice a day for two weeks, then 1000 mg by mouth 
twice a day for ten weeks. 
– for patients greater than 40 kg: 500 mg by mouth twice a day for two weeks, then 1000 mg by 
mouth twice a day for two weeks, then 1000 mg by mouth each morning and 2000 mg by mouth 
every afternoon for eight weeks.  
– Dosages will be increased based on tolerability and the monitoring of any potential GI side 
effects as determined by the investigator. 
 
Dosing Regimen for <40kg group (BID dosing throughout study)  
Week 0 (Visit 1) – Week 2 (Visit 2):  
1000mg/day (500mg po qam and 500mg po qpm)  
Week 2 (V2) – Week 12 (V5): 
 
2000mg/day (1000mg po qam and 1000mg po qpm) 
 
Dosing Regimen for >40kg group (BID dosing throughout study)  
Week 0 (Visit 1) – Week 2 (Visit 2):  
1000mg/day (500mg po qam and 500mg po qpm)  
Week 2 (V2) – Week 4 (V3):  
 
2000mg/day (1000mg po qam and 1000mg po qpm) 
Week 4 (V3) – Week 12 (V5):  
 
3000mg/day (1000mg po qam and 2000mg po qpm) 
 
Maximum Amount of Study Drug Needed  
V1: 1000mg = 2 (500mg) pills x 14 days = 28 pills  
V2: 2000mg = 4 (500mg) pills x 14 days = 56 pills  
V3: 3000mg = 6 (500mg) pills x 28 days = 168 pills  
V4: 3000mg = 6 (500mg) pills x 28 days = 168 pills  
V5: none dispensed (end of study) total per subject: 420 pills 
TOTAL PILLS for 24 subjects = ~10,080pills maximum (based on recruiting patients >40kg) 
 
 
SUMMARY OF DATA COLLECTION AND STUDY PROCEDURES BY VISIT 
 
 
Visit 

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Screening 
1 
2 
3 
4 
5 
6 
Consent Forms: 
X 
 
 
 
 
 
 
Medical Procedures on Child 
Blood draw 
 
X 
 
 
 
 
X 
Heart rate 
 
X 
X 
X 
X 
X 
X 
Blood pressure 
 
X 
X 
X 
X 
X 
X 
Parent Reports on Child 
Aberrant Child 
Checklist 
X 
X 
X 
X 
X 
X 
X 
Social 
responsiveness 
Scale 
X 
X 
X 
X 
X 
X 
X 
Clinical Global 
Impression Scale 
X 
X 
X 
X 
X 
X 
X 
PRAS ASD 
X 
 
 
 
 
 
X 
Vineland-3 
X 
 
 
 
 
 
X 
Side effects 
checklist 
 
X 
X 
X 
X 
X 
X 
 
 

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Subject Recruitment  
All potential subjects for this study will be recruited through the Child and Adolescent Psychiatry 
Department Neurodevelopmental clinic and by Neuropsychological referral.  
 
Subjects:  
Inclusion criteria:  
1) Boys and girls ages 4-17;  
2) Current diagnosis of Autism Spectrum Disorder as determined by criteria in DSM-5 
3) Parents of Children ages 4-17 with a current diagnosis of Autism Spectrum Disorder as 
determined by criteria in DSM-5 
 
Exclusion criteria:  
1) Unstable medical illness or clinically significant abnormalities on physical examination;  
2) History of seizures;  
3) History of Hematological disorders;  
4) Myocardial infarction within 6 months;  
5) Current pregnancy or lactation, or inadequate contraception in girls of childbearing potential;  
6) Current or recent (past 3 months) DSM-5 substance abuse or dependence;  
7) Illegal substance use within 2 weeks of study initiation;  
8) Previous treatment with Glutathione;  
9) Current treatment with N-acetylcysteine, milk thistle, Vitamin C, Vitamin B, Grape Seed 
Extract, Amino Acids, or Zinc (known to interfere with Glutathione level) 
11) Current treatment with Dextromethorphan, D-cycloserine, Amantadine, Memantine, 
Lamotrigine or Riluzole (known to have glutamatergic properties) 
12) Asthma (given this is rare, but possible worsening of asthma symptoms due to Glutathione) 
 
Assessments:  
Existing Data 
Before beginning Glutathione, all subjects will have a psychiatric, medical, and family history 
evaluation completed as part of their standard clinical care.  
 
New Data Collection 
At the screening visit, all subjects will receive a physical examination. For all subjects, at 
the initial visit, blood work will be obtained for a baseline Glutathione level. Glutathione level will 
also be obtained at study completion. Total amount of blood to be obtained will be 4 teaspoons. PI 
will obtain neuropsychological evaluations report if it is available.  
The following instruments will be completed at each visit throughout the study: 1) Aberrant 
Behavior Checklist [ABC], a 58-item parent report subjective scale used for assessing problem 
behavior in subjects with intellectual and developmental disabilities; 2) Social Responsiveness 
Scale [SRS], a 65-item parent report questionnaire designed to use with children ages 4-17, 
providing age- and gender-referenced ranges and assessing social deficits pertaining to autism and 

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other domains; and 3) the Clinical Global Impression scale [CGI]; 7 point scale to measure 
symptom severity. Also, readings of patients’ heart rates and blood pressures will be obtained; and 
assessment of side effects will be done at each visit. 
Note on ABC and SRS:  
The ABC is a standardized scale used for assessing problem behaviors in subjects with 
developmental and intellectual disabilities. The items are divided into five subscales: 1) 
Irritability; 2) Lethargy/Social Withdrawal; 3) Stereotypic Behavior; 4) Hyperactivity; and 
5) Inappropriate Speech. High scores are indicative of severe behavioral symptoms. The 
ABC will be used to track the subjects’ problem behaviors and difficulties over the course 
of the study.  The severity of the subjects’ problem behaviors must be assessed and 
examined its relationship with a baseline blood level of glutathione. In addition, by using 
this scale, we can observe the overall effect of oral Glutathione on problem behaviors, 
especially comparing the end results to the baseline.  
 
The SRS is being used for the same purpose, although it is being used for secondary 
outcome measures. It is a parent report questionnaire designed for use with children aged 
4-17 and provides age- and gender-referenced, as well as raw scores for the following 
domains: total score (reflecting severity of social deficits pertaining to autism); receptive, 
expressive, cognitive and motivational aspects of social behavior; and autistic 
preoccupations. High scores are also indicative of severe behavioral symptoms. 
 
Note on additional measures: 
Clinical Global Impression (CGI) (Severity) scale will be used by the clinician to assess 
overall severity of problem behaviors in Autism Spectrum Disorder subjects at baseline. 
This will be used as a secondary outcome measure over the course of the study and relates 
to Aim #1. 
 
 

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Study Population/Sample Size  
24 male and female outpatients aged 4-17 with a primary diagnosis of Autism Spectrum Disorder. 
 
-Note on sample size: The sample size is low due to this being a pilot feasibility study and 
the primary aim of the study is not to examine the effectiveness of oral glutathione as a 
treatment but rather examine the functionality of providing oral glutathione on the blood 
level, the data will likely be used for future studies on treatment or grant applications. There 
also is no external sponsor, therefore internal funding limits the number of subjects.  
Participant Exclusion:  
For this study, children under the age of 4 and adults over the age of 17 will be excluded. The 
rationale for this is that the study population of interest are pediatric subjects in the children and 
adolescent age ranges.  
 
Data Analysis:  
Primary analyses will be intent-to-treat with last observation carried forward. Changes in the blood 
level of glutathione will be the primary outcome measure. For both primary and secondary 
measures, random regression analysis will be conducted using the MIXREG program 
Demographic and baseline visit characteristics of the Glutathione treatment patients will be 
documented. Primary and secondary measures will be examined using analysis of variance 
modeling analyses (SPSS). The difference in the overall level of posttreatment values, the main 
effect for treatment, will be the test of primary interest. Analyses will be performed on all available 
data by the use of an intention-to-treat population (last observation carried forward). All patients 
who return for at least 1 follow-up visit will be included in the intention-to-treat population. Effect 
sizes will also calculated using the Cohen effect size index d. A d of 0.2 is considered a small 
effect size, 0.5 is medium, and 0.8 is large. Partial eta squared (η2), which is the proportion of the 
effect + error variance that is attributable to the effect, will be calculated. Interpretation of η2 is 
that greater than 0.2 is a large effect size, greater than 0.1 is a medium effect size, and greater than 
0.05 is a small effect size. 
The CGI severity scale will be used to assess clinically significant change after treatment. 
For someone to have a clinically significant change, her or his final CGI severity score had to be 
less than the cutoff score, and her or his change from baseline had to be greater than the reliable 
change index 
The study will end once 24 subjects have been recruited and have completed the study 
visits. At this point, the data analysis will then begin. 
 
Potential Risks and Benefits: 
The study supplement Glutathione may help to decrease problem behaviors and irritability that can 
be seen in children with Autism Spectrum Disorder. It is also postulated that the information 
learned from this study will benefit other individuals and children and adolescents with problem 
behaviors related to Autism Spectrum Disorder in the future. 
 

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Most supplements can cause side effects, but are typically safe. Side effects are usually reversible 
when the supplement is stopped. The most common side effects associated with Glutathione 
include; upset stomach, diarrhea, dizziness, dry mouth or headaches and sometimes, but rarely a 
skin rash, skin flushing or respiratory distress can be seen. As such, patients with asthma or 
allergies that have resulted in respiratory distress in the past will not be included in this study to 
ensure safety. 
 
Safety Assessments:  
Safety assessments (sitting blood pressure, heart rate, adverse effects, and concomitant 
medications) will be documented at each visit. In terms of vital signs, those subjects with abnormal 
blood pressures will be assessed for symptoms of hypo- or hypertension. Asymptomatic subjects 
will be evaluated each visit for changes in vital signs. In the case of hypertensive emergencies (BP 
greater than 210/120), appropriate referral to the emergency room will be made. In the case of 
hypotension (BP less than 90/60), participants will be evaluated for symptoms of hypotension and 
if symptomatic, appropriate interventions will be made. Subjects who endorse suicidal thoughts at 
any time during the study will be removed from the study and appropriate clinical intervention 
(e.g. hospitalization) will be arranged.  
 
At the initial visit, blood work will be obtained for a baseline Glutathione level and an end 
point Glutathione level will also be obtained at study completion. Assessment of side effects will 
be done at each visit. Participants will be advised concerning the risk of allergic reactions to 
Glutathione: skin rash, skin flushing, drop in blood pressure, irregular heartbeat, and respiratory 
distress. Although is it extremely rare but participants will be advised to visit an emergency room 
for symptoms suggestive of anaphylactoid reactions. 
 
Adverse Event Reporting:  
Unanticipated (unexpected) problems/events, those that are not already described as potential risks 
in the consent form, or not part of an underlying disease, will be reported to the IRB within 10 
business days. The same will be done for serious problems/events and, in the opinion of the 
investigator, are possibly, probably or definitely related to the research procedures. A follow-up 
report will also be submitted to the IRB with any documentation related to a previously submitted 
adverse event. 
 
Procedure:  
Twenty-four (24) subjects will be recruited to receive oral Glutathione reduced in this open-label 
trial. All parents of the subjects will be provided with informed consent forms and verbal assent 
from subjects > 6 years of age will be obtained after an age appropriate version of the informed 
consent forms are read to them. The Child Psychiatry clinicians/nurses, patients and parents will 
be aware of the medication name and dosing schedules as indicated. The clinic nurse will obtain 
vitals at each visit.  Glutathione will be the only study medication dispensed to subjects for this 
study. Subjects will be told to take the first dose of study medication after breakfast each day and 
the second dose in the evening after dinner. Subjects will be told to monitor and report any side 
effects experienced to the study staff at each visit or immediately if they wish (via one of the 

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multiple telephone numbers for the study physicians and study staff given to them at each visit). 
Psychotherapy of any form (including cognitive-behavioral therapy) will be reported to the 
PI during the study. 
Subjects will be evaluated with the Aberrant Behavior Checklist (ABC), Social 
Responsiveness Scale (SRS) and CGI at screening and at each visit for the remainder of the study. 
Medication side effects will be evaluated at each study visit. A tablet count will be kept for each 
dose of medication taken to monitor adherence. 
 
Dosing Regimen for Glutathione  
The proposed dose range for Glutathione in this study will be 1000mg-3000mg/day based on the 
subject’s weight. A previous observational study of children with Cystic Fibrosis showed that this 
population responded well and were able to absorb Glutathione with a daily dose of 65mg/kg (10).  
As Cystic Fibrosis is a condition that typically results in malabsorption of fats and other 
products, it is our thought that subjects with normal gastrointestinal (GI) systems may also absorb 
Glutathione effectively and at a slightly lower milligram starting dose, which is proposed to be 
~32.5mg/kg. No adverse events were noted in the above mentioned study; however, to ensure 
tolerability in subjects with normal functioning GI systems, starting at a low dose and titrating up 
accordingly will be the most conservative and safe method. 
 
Timeline: 
Recruitment for the proposed study will begin as soon as the study drug is available and IRB 
approval is confirmed. It is anticipated that all subjects would be entered into the study within 
approximately 6 months, and that the study would be completed within 12 months of initiation.  
 
Ethical Considerations: 
This study will be conducted according to US and international standards of Good Clinical Practice 
(FDA regulations 21 CFR 312 for IND studies and FDA guidance E6) for all studies. Applicable 
government regulations and University of Chicago research policies and procedures will also be 
followed. 
This protocol and any amendments will be submitted to the University of Chicago 
Institutional Review Board (IRB) for formal approval to conduct the study. The decision of the 
IRB concerning the conduct of the study will be made in writing to the investigator. 
Parents of all subjects for this study will be provided a consent form describing this study 
and providing sufficient information for parents to make an informed decision about their 
participation in this study. This consent form will be submitted with the protocol for review and 
approval by the IRB. The formal consent of parents of subjects, using the IRB- approved consent 
form, will be obtained before that subject is submitted to any study procedure. This consent form 
must be signed by the parents of subjects or legally acceptable surrogate, and the investigator-
designated research professional obtaining the consent. 
 
Data Handling and Record Keeping  

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All subject data will be maintained by the study personnel under the supervision of the principal 
investigator. All subjects will be seen in the Department of Child and Adolescent Psychiatry at the 
University of Chicago Medical Center at 5841 South Maryland Avenue. Patient folders will be 
kept in a locked cabinet only accessible to research staff. 
 
Finance and Insurance  
All research will be paid for by the internal departmental funds of the Department of Child and 
Adolescent Psychiatry at the University of Chicago. 
In the event that this research activity results in an injury, treatment will be available, 
including first aid, emergency treatment and follow-up care as needed. Care for such injuries will 
be billed in the ordinary manner to the subjects’ insurance company. If the parents of subjects 
believe that they have suffered a research related injury, they will be instructed to inform the 
principal investigator immediately. 
 
Compensation 
Participation in this study will be completely voluntary and no monetary compensation will be 
offered. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
REFERENCES 

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1. American Psychiatric Association: Diagnostic and Statistical Manual of Mental Disorders: 
Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. Arlington, VA: 
American Psychiatric Association, 2013. 
2. Baio J, Wiggins L, Christensen DL, et al. Prevalence of Autism Spectrum Disorder Among 
Children Aged 8 Years — Autism and Developmental Disabilities Monitoring Network, 
11 Sites, United States, 2014. MMWR Surveill Summ 2018;67(No. SS-6):1–23 
3. Ghanizadeh A, Akhondzadeh S, Hormozi M, Makarem A, Abotorabi-Zarchi M, 
Firoozabadi A. Glutathione-Related Factors and Oxidative Stress in Autism, A Review: 
Current Medicinal Chemistry 2012;19:4000-4005   
4. Harden AY, Fung LK, Libove RA, Obukhanych TV, Nair S, Herzenberg LA, Frazier TW, 
Tirouvanziam R. A Randomized Controlled Pilot Trial of Oral N-Acetylcysteine in 
Children with Autism. Biol Psychiatry 2012;71:956-961  
5. Kern JK, Geier DA, Adams JB, Garver CR, Audhya T, Geier MR. A Clinical Trial of 
Glutathione Supplementation in Autism Spectrum Disorders. Med Sci Monet 2011; 
17(12):CR677-682 
6. Lu SC. Glutathione Synthesis. Biochem Biophys Acta 2013;1830(5):3143-3153 
7. Main PA, Angley MT, O’Doherty CE, Thomas P, Fenech M. The Potential Role of the 
Antioxidant and Detoxification Properties of Glutathione in Autism Spectrum Disorders: 
A systematic Review and Meta-Analysis 
8. Mcguire K et al. Irritability and Problem Behavior in Autism Spectrum Disorder: A 
Practice Pathway for Pediatric Primary Care. Pediatrics 2016;137 
9. Rushworth GF, Megson IL. Existing and Potential Therapeutic Uses for N-acetylcysteine: 
The need for Conversion to Intracellular Glutathione for Antioxidant Benefits. Pharmacol 
Ther 2014;141(2):150-9 
10. Visca A, Bishop CT, Hilton S, Hudson VM. Oral Reduced Glutathione Improves Growth 
in Pediatric Cystic Fibrosis Patients. Journal of Pediatric Gastroenterology and Nutrition 
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11. Wink LK, Adams R, Wang Z, Klaunig JE, Plawecki MK, Posey DJ, McDougle CJ, 
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with Autism Spectrum Disorder. Molecular Autism 2016;7:26 
 
 
 
 
 
 
 
 
 
Appendix A 

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Diagnostic Criteria for Autism Spectrum Disorder (1) 
The following diagnostic criteria for ASD will be utilized. Criteria are defined by the Diagnostic 
and Statistical Manual for Mental Disorders (DSM-5) for a diagnosis of Autism Spectrum Disorder 
(1): 
A. Persistent deficits in social communication and social interaction across multiple 
contexts, as manifested by the following, currently or by history (examples are illustrative, not 
exhaustive, see text): 
1.       Deficits in social-emotional reciprocity, ranging, for example, from abnormal social approach 
and failure of normal back-and-forth conversation; to reduced sharing of interests, emotions, or 
affect; to failure to initiate or respond to social interactions. 
2.       Deficits in nonverbal communicative behaviors used for social interaction, ranging, for 
example, from poorly integrated verbal and nonverbal communication; to abnormalities in eye 
contact and body language or deficits in understanding and use of gestures; to a total lack of facial 
expressions and nonverbal communication. 
3.       Deficits in developing, maintaining, and understanding relationships, ranging, for example, 
from difficulties adjusting behavior to suit various social contexts; to difficulties in  sharing 
imaginative play or in making friends; to absence of interest in peers. 
Specify current severity: Severity is based on social communication impairments and restricted 
repetitive patterns of behavior.  
B. Restricted, repetitive patterns of behavior, interests, or activities, as manifested by at 
least two of the following, currently or by history (examples are illustrative, not exhaustive; see 
text): 
1.       Stereotyped or repetitive motor movements, use of objects, or speech (e.g., simple motor 
stereotypies, lining up toys or flipping objects, echolalia, idiosyncratic phrases). 
2.       Insistence on sameness, inflexible adherence to routines, or ritualized patterns or verbal 
nonverbal behavior (e.g., extreme distress at small changes, difficulties with transitions, rigid 
thinking patterns, greeting rituals, need to take same route or eat food every day). 
3.       Highly restricted, fixated interests that are abnormal in intensity or focus (e.g., strong 
attachment to or preoccupation with unusual objects, excessively circumscribed or perseverative 
interest). 
4.       Hyper- or hyporeactivity to sensory input or unusual interests in sensory aspects of the 
environment (e.g., apparent indifference to pain/temperature, adverse response to specific sounds 
or textures, excessive smelling or touching of objects, visual fascination with lights or movement). 
Specify current severity: Severity is based on social communication impairments and restricted, 
repetitive patterns of behavior.  
C. Symptoms must be present in the early developmental period (but may not become fully 
manifest until social demands exceed limited capacities or may be masked by learned strategies in 
later life). 
D. Symptoms cause clinically significant impairment in social, occupational, or other 
important areas of current functioning. 
E. These disturbances are not better explained by intellectual disability (intellectual 
developmental disorder) or global developmental delay. Intellectual disability and autism 
spectrum disorder frequently co-occur; to make comorbid diagnoses of autism spectrum disorder 

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and intellectual disability, social communication should be below that expected for general  
developmental level. 
Note: Individuals with a well-established DSM-IV diagnosis of autistic disorder, 
Asperger’s disorder, or pervasive developmental disorder not otherwise specified should be given 
the diagnosis of autism spectrum disorder. Individuals who have marked deficits in social 
communication, but whose symptoms do not otherwise meet criteria for autism spectrum disorder, 
should be evaluated for social (pragmatic) communication disorder. 
Specify if: 
- With or without accompanying intellectual impairment 
- With or without accompanying language impairment 
- Associated with a known medical or genetic condition or environmental factor 
- Associated with another neurodevelopmental, mental, or behavioral disorder 
- With catatonia