Clinical trial NCT03835117
Conditions: Autism Spectrum Disorder
Keywords: Mitochondrial Dysfunction
Sponsor: Rossignol Medical Center
Start date: 2020-02-01; Completion date: 2022-07-31
Enrollment: 16 participants; Allocation: RANDOMIZED; Intervention model: CROSSOVER

Brief summary:
The objective of this study is to evaluate the metabolic effects of a comprehensive wide-spectrum supplement for children with ASD to determine whether it physiologically targets mitochondrial pathways known to be abnormal in children with ASD.The intervention is a commonly used wide-spectrum nutritional supplement, which is theoretically designed to normalize mitochondrial function. The investigators aim to determine if the supplement does have the hypothesized effect on physiology in individuals with ASD. The investigator will enroll up to 50 children, aged 4 to 14 years of age with confirmed ASD and mitochondrial dysfunction, and participation will last 26 weeks.

Detailed description:
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder often with life-long consequences that affects young children during critical developmental periods. The Centers for Disease Control estimates that ASD affects as many as 17 per 1000 children (1 in 59) in the United States suggesting that the prevalence is higher than previous estimates.Despite the dramatic rise in the detected prevalence of ASD over the past two decades, there is no effective medical treatment for core ASD symptoms (social communication and repetitive behavior), the closely associated problem of language impairment, or the underlying pathophysiology of ASD. Currently, the only accepted treatment for core ASD symptoms is behavior therapy, which may entail intensive one-on-one treatment over several years.

The primary aims of this study are to evaluate the effect of a wide-spectrum nutritional supplement on mitochondrial function in individuals with ASD. Participants entered into the trial will have abnormalities in mitochondrial function that are known to be associated with ASD (approximately 50+% of children with ASD) but are not diagnostic of mitochondrial disease. The investigators hypothesize that nutritional supplements designed for children with ASD have a physiological action of normalizing mitochondrial function and cellular physiology throughout the body.

To test whether the targeted nutritional supplement is superior to placebo, the investigators will study 50 children, between the ages of 4 years to 14 years, with confirmed ASD and known abnormal variations in mitochondrial at baseline. Participants will be randomly assigned to receive active treatment or placebo for 12-weeks under double-blind conditions and at the end of the 12 weeks switch to the opposite condition after a 2-week wash out period. Mitochondrial function will be measured at baseline and after each treatment arm in order to determine if the supplement positively influences cellular biochemistry. The investigator will also evaluate the effectiveness of the supplement on core and associated ASD symptoms using several behaviors assessments.

Arms: arm 1: The Wide-spectrum nutritional supplement used will be a combination of NeuroNeeds: SpectrumNeeds and QNeeds. Weight based dosing will be used. The daily serving size will be divided into two oral daily doses in the form of a powder which can be mixed into liquid or food. Together, there are 34 different dietary supplements in the products. Except for ubiquinol, all of these nutrients are provided in a powder form in SpectrumNeeds. Ubiquinol is provided separately in QNeeds gel capsules. These capsules can be swallowed whole, or cut with scissors and the contents squeezed out and added to SpectrumNeeds just before ingestion. arm 2: Participants randomized to receive placebo will take placebo in an oral form divided into powder and a gel capsule in the same manner as treatment. For the second phase of the cross over, participants will be part of the opposite group they were assigned to in Phase I (Placebo or Treatment). Quantities for placebo or treatment will match across phases for each subject, utilizing the same weight based dosing.
Interventions: intervention 1: Wide-spectrum nutritional supplement intervention 2: Placebo
Intervention descriptions: intervention 1: Comprehensive powder with 33 dietary supplements and 1 dietary supplement via gel capsule. intervention 2: Inactive placebo comparator

Protocol documents:
Prot_SAP_000.pdf: 
Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
1 
 
 
Synopsis 
Protocol Title 
Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in 
Children with Autism Spectrum Disorder (ASD) 
IRB 
WCG Institutional Review Board 
Protocol Ver 
Wednesday, August 3, 2022 
Sponsor 
Richard E Frye, Rossignol Medical Center, Phoenix AZ 
IND # / FDA 
IND 142751; FDA Division of Psychiatry; Protocol Version 1.4 
NCT ID 
NCT03835117 
Principal 
Investigator 
Richard E. Frye, MD, PhD, Rossignol Medical Center, 4045 E Union Hills Drive,  
Suite 116, Phoenix, Arizona 85050 
Site- Investigator 
Christopher Smith, PhD, Southwestern Autism Resource and Research Center 
(SARRC), Phoenix AZ 
Co-Investigators 
1. Richard Boles, MD, Director of CNNH NeuroGenomics Program, Center for 
Neurological and Neurodevelopment Health, Voorhees, NJ.    
2. Dr. Richard Tayrien D.O. (SARRC), Phoenix AZ 
Clinical Sites 
1. SARRC, Phoenix, AZ 
Laboratory Sites 
1. Laboratory of Dr. Edward Quadros, State University of New York 
Downstate—450 Clarkson Ave, Brooklyn, NY 
2. Religen Inc, Plymouth Meeting, PA 
3. Laboratory of Dr Arora, Icahn School of Medicine at Mount Sinai, NY NY 
4. Laboratory of Dr. Madeleine Cunningham at the University of Oklahoma 
Health Sciences Center in Oklahoma City, OK 
5. 3billion Laboratory in Seoul, South Korea 
6. Iliad Inc, Plymouth Meeting, PA 
Study Design 
Prospective Randomized 24-week Double-Blind Placebo-Controlled Cross-over 
Objectives 
To evaluate the metabolic effects of a comprehensive wide-spectrum supplement 
for children with ASD to determine whether it physiologically targets mitochondrial 
pathways known to be abnormal in children with ASD.  
Participants  
Up to 50 children, aged 2 years 6 months to 17 years 3 months with confirmed 
ASD and mitochondrial dysfunction. Participants will have moderate disease 
severity (clinic global impression of 4 or above) and stable behavioral, education 
and medical treatment. Those with severe intellectual impairment, disruptive 
behavior or medical conditions such as epilepsy will be excluded.    
Investigational 
Product 
Weight adjusted wide-spectrum oral nutritional supplement powder and gel 
capsules that targets mitochondrial support produced by NeuroNeeds (Old Lyme, 
CT) taken in two divided doses vs oral placebos. 
Safety 
Physical examination, safety labs, Adverse Event Reporting, safety monitoring 
Statistical 
Methods 
The primary analysis will be intent-to-treat, defined as all randomized subjects. 
Mixed-model regression will determine whether blinded treatment produces a 
greater normalization in mitochondrial function over the 12-week period as 
compared to placebo treatment over the same time period. Similar analysis will 
test secondary outcome measures. Assuming a medium effect size of 0.5, the 
study provides an 80% power assuming a 0.01 alpha (allowing correction for 
multiple primary end-points). Secondary outcome measures will include the 
Improvement item of the Clinical Global Impression Scale and behavioral scales 
such as the Aberrant Behavior Checklist.  Multivariable logistic regression will be 
used to select biomarkers that can significantly predict treatment response. 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
2 
 
Funding  
Turnabout for Autism (Private Foundation), Pleasanton CA 
 
Contents 
Synopsis................................................................................................................................................. 1 
1. ABSTRACT ........................................................................................................................................ 3 
2. BACKGROUND................................................................................................................................. 3 
3. An Integrated Approach to Understanding Autism Spectrum Disorder .......................... 4 
4. SPECIFIC AIMS ................................................................................................................................. 6 
5. RESEARCH PLAN ........................................................................................................................... 7 
6. DATA MANGEMENT AND STATISTICS ................................................................................... 13 
7. HUMAN SUBJECTS CONCERNS............................................................................................... 14 
8. REFERENCES................................................................................................................................. 19 
 
 
 
 
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
3 
 
1. ABSTRACT 
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder often with life-long 
consequences that affects young children during critical developmental periods. The Centers for 
Disease Control estimates that ASD affects as many as 17 per 1000 children (1 in 59) in the United 
States suggesting that the prevalence is higher than previous estimates.1 Despite the dramatic rise in 
the detected prevalence of ASD over the past two decades, there is no effective medical treatment for 
core ASD symptoms (social communication and repetitive behavior), the closely associated problem 
of language impairment, or the underlying pathophysiology of ASD. Currently, the only accepted 
treatment for core ASD symptoms is behavior therapy, which may entail intensive one-on-one 
treatment over several years. Such therapy is inconsistently covered by medical insurance and may 
not be available in special education settings, thereby limiting its application. The de facto standard-
of-care for treating ASD is a combination of educational and behavioral interventions.  
 
Although interventions such as speech and occupational therapy may help modify neural pathways, 
they may not address the pathophysiology that underlies ASD. Despite these interventions, most 
children with ASD have incomplete recovery and need for long-term supportive care. Thus, a safe, 
effective and well-tolerated medical treatment that improves core and closely associated ASD 
symptoms, addresses the underlying physiology associated with ASD, and augments ongoing 
behavioral and educational interventions would have disease-modifying potential. To better 
understand if treatments commonly used in children with ASD are indeed normalizing physiological 
disturbances associated with ASD we aim to determine if a commonly used wide-spectrum nutritional 
supplement theoretically designed to normalize mitochondrial function does have the hypothesized 
effect of physiology in individuals with ASD. 
 
The primary aims of this study are to evaluate the effect of a wide-spectrum nutritional supplement on 
mitochondrial function in individuals with ASD. Participants entered into the trial will have 
abnormalities in mitochondrial function that are associated with ASD (such abnormalities are found in 
approximately 50% of children with ASD) but are not diagnostic of mitochondrial disease. Specifically, 
children with mitochondrial disease (approximately 5% of the ASD population), which is a very severe 
disorder, will be excluded from the study so they can pursue more intensive appropriate treatment. As 
we believe 50% of children with ASD have some form of atypical variations in mitochondrial function, 
we will target these children to improve the function of their mitochondria. We hypothesizes that a 
nutritional supplement designed for children with ASD will have a physiological action of normalizing 
mitochondrial function and cellular physiology throughout the body. To test whether the targeted 
nutritional supplement is superior to placebo, we will study 50 children, between the ages of 2 years 6 
months and 17 years 3 months, with confirmed ASD and known abnormal variations in mitochondrial 
at baseline. Participants will be randomly assigned to receive active treatment or placebo for 12-
weeks under double-blind conditions and at the end of the 12 weeks switch to the opposite treatment 
arm. Mitochondrial function will be measured at baseline and after each treatment arm in order to 
determine if the supplement positively influences cellular biochemistry. 
2. BACKGROUND 
ASD is a heterogeneous neurodevelopmental disorder often with life-long consequences that affects 
young children during critical times in their development.2 The latest estimates from the Center for 
Disease Control estimates an ASD affects as many as 17 per 1000 children (1 in 59) in the United 
States with recently estimation demonstrating that the provenience continues to rise.1 ASD is defined 
by impairments in social communication as well as the presence of restrictive interests and repetitive 
behaviors2. ASD is frequently associated with co-occurring problems  such as gastrointestinal 
abnormalities3, seizures4, attention deficits5, anxiety6 and allergies7 -  to name a few. 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
4 
 
Despite the fact that the prevalence of ASD has dramatically increased over the past two decades, 
there currently is no effective medical treatment for children with ASD. A recent multicenter study 
showed that optimal outcomes can be achieved in a subset of ASD children.8 Moreover, a study that 
analyzed the annual change in ASD symptoms in 6,975 children showed that some children with ASD 
can substantially improve.9 However, these studies did not address whether particular therapies were 
associated with better outcomes. In addition, the findings suggest that only a minority of children with 
ASD show substantial improvement over time, underscoring the need for more effective treatments. 
 
Currently the only well-accepted treatment for core ASD symptoms is behavior therapy such as 
Applied Behavioral Analysis (ABA) and Early Intensive Behavioral Intervention. However, such 
therapy requires near full-time engagement with a one-on-one therapist over several years.  The 
evidence for early intervention rests primarily on single-subject design studies with few randomized 
controlled studies and these interventions rarely result in complete recovery.12 Moreover, access to 
such intensive treatments is often limited and may only be available in highly specialized centers.13 
Behavior therapy is inconsistently covered by medical insurance and not uniformly available in the 
education system.10,11 Thus, the de facto standard-of-care for treating ASD is a combination of 
educational and allied health therapies (speech and occupational therapies) in various settings. 
Although these interventions may promote development in children with ASD, they may not address 
the underlying pathophysiological abnormalities in ASD.  Most individuals with ASD require life-long 
supportive care and do not achieve independent living in adulthood.12 
 
Evidence-based medical treatments for ASD are limited. There is no United States (US) Food and 
Drug Administration (FDA) approved medical therapy that addresses core ASD symptoms or the 
pathophysiological processes that underlie ASD.13 The two FDA approved drugs, both atypical 
antipsychotics, are indicated for serious behavioral problems in children with ASD. Although these 
drugs, risperidone and aripiprazole, may improve social interactions as a secondary benefit,18 the 
detrimental effects on lipid and glucose metabolism and body weight are well established and may 
develop quickly (within 12 weeks).14-16 These metabolic changes increase the risk of cardiovascular 
disease and type-2 diabetes. 17 17 17 Although the incidence is low, tardive dyskinesia, a potentially 
permanent movement disorder, is an additional risk.18 Thus, there is a pressing need for safe 
medications that affect target pathophysiological processes and treat core ASD symptoms. 
 
Several medications are used off-label to treat associated, but not core, ASD symptoms. These 
medications do not address underlying pathophysiological abnormalities. A systematic review of 
attention-deficit hyperactivity disorder (ADHD) treatments for ASD highlights the complicated nature of 
medication treatment due to the diverse nature of children with ASD and associated adverse effects 
(AEs).19 A recent Cochrane review found no evidence for efficacy of selective serotonin reuptake 
inhibitors in ASD.23 Although studies show positive effects of methylphenidate and extended-release 
guanfacine on hyperactivity, these medications did not improve core features of ASD and the AE 
burden was higher than found in children with ADHD.24,25 Pharmaceutical companies have developed 
medications targeting excitatory-inhibitory neurotransmission imbalances identified in animal models 
of ASD20, but the first of these compounds  failed to demonstrate efficacy in a double-blind placebo-
controlled (DBPC) trial, leading to termination of the research program.21  
3. An Integrated Approach to Understanding Autism Spectrum Disorder 
Our novel approach to treating ASD is to address the underlying physiological abnormalities 
associated with ASD as we hypothesize that these physiological abnormalities are driving abnormal 
behavior and cognition. Of note, ASD is associated with physiological disturbances including 
abnormal redox and mitochondrial metabolism. In fact, between 5%-80% of children with ASD 
demonstrate evidence of mitochondrial dysfunction and 5% of children with ASD appear to have 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
5 
 
mitochondrial disease.22-24 This is in comparison to the general population where mitochondrial 
disease is believed to affect less than 0.1% of the population.25   
 
Repeated studies have suggested that genetic defects are found only in a minority of children with 
ASD, including a minority of children with ASD with mitochondrial disease22 and many case reports 
and series have described only moderate, rather than severe, deficiencies in the electron transport 
chain (ETC) activity in those with ASD and mitochondrial disease.24,26 Perhaps more striking is the 
fact that ETC activity in muscle,26,27 skin,24 buccal epithelium28-30 and brain 31 has been documented to 
be significantly increased, rather than decreased, in some individuals with ASD. This is consistent 
with our in vitro data showing elevated mitochondrial activity in lymphoblastoid cell lines (LCLs) 
derived from children with ASD.32-38 We recently demonstrated that this alteration in mitochondrial 
function is associated with more severe repetitive behaviors.35 Thus, the current data points to 
possible non-genetic defects or changes causing abnormal mitochondrial function as well as unique 
types of mitochondrial dysfunction in individuals with ASD.24,27  
 
Mitochondrial dysfunction has profound significance for health. Mitochondria are essential for a wide 
range of functions in almost every cell in our body. Best known for their role in the production of 
adenosine triphosphate (ATP), mitochondria are also intimately involved in other essential cellular 
functions such as calcium buffering, redox regulation, apoptosis and inflammation and have a role in 
non-energy producing metabolic pathways, such as the urea cycle and porphyrin pathway. Additionally, 
ATP produced by the mitochondria is essential for many cellular systems. Thus, abnormal mitochondrial 
function can adversely affect a wide range of cellular systems. Mitochondrial dysfunction is recognized 
to contribute to the pathophysiology of many common diseases, especially those thought to have a 
substantial environmental component, including psychiatric diseases,39-42 neurodegenerative 
disorders,43 persistent systemic inflammation,44 cardiac disease,45 cancer46 and diabetes.47 
Abnormalities in mitochondrial function are closely associated with neurodevelopmental disorders, 
including genetic syndromes closely associated with ASD including phosphatase and tensin homolog 
(PTEN) and WDR45 mutations,48,49 septo-optic dysplasia50 and Rett51-53, Phelan-McDermid,54 15q11-
q13 duplication,55,56 Angelman57 and Down58,59 syndrome.  
 
Most significantly, there is evidence that nutritional supplement used in children with ASD target the 
mitochondrial. Several studies have demonstrated that nutritional supplements, such as carnitine,  
nicotinamide adenine dinucleotide (NAD), ubiquinol and ubiquinone designed to treat individuals with 
mitochondrial disease may be helpful in children with ASD.21 Two DBPC studies demonstrated 
improvement in ASD symptoms with carnitine supplementation60 with some improvements directly 
related to the change in blood carnitine levels.61  In another study, NAD and ribose appeared to 
improve metabolic biomarkers in children with ASD and symptoms of mitochondrial dysfunction.62 
Two studies have reported behavioral improvements in children with ASD using ubiquinol63 and 
ubiquinone.64 While these aforementioned studies did not specifically target individuals with 
mitochondrial dysfunction, one recent study examined the behavioral effects of a customized 
mitochondrial supplement in ASD children with mitochondrial dysfunction. Legido et al65 in an open-
label study treated 11 children with ASD and abnormal Complex 1 and/or Complex 4 activity, as 
measured by the buccal swab method, with a mitochondrial cocktail containing carnitine, coenzyme 
Q10 and alpha-lipoic acid. Three months of treatment improved the index of mitochondrial function 
and improved several behavior scales including lethargy and inappropriate speech subscales from the 
Aberrant Behavior Checklist (ABC). Three months after withdrawal of the treatment the lethargy and 
inappropriate speech subscales significantly worsened. 
 
Thus, we aim to build on this previous evidence in order to demonstrate that a mitochondrial targeted 
nutritional supplement commonly given to children with ASD indeed positively affect mitochondrial 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
6 
 
function. Previous studies have been low quality because they were either open-label or did not 
measure physiological response of the treatment. We wish to perform a rigorous study that targets the 
subset of children with ASD with abnormal variations in mitochondrial function associated with ASD in 
order to increase the scientific evidence to understand how the physiological mechanisms which 
nutritional supplements may affect children with ASD. 
4. SPECIFIC AIMS 
Primary Aim: To determine if a wide-spectrum nutritional supplement can influence 
mitochondrial activity in 50 individuals with ASD and signs and symptoms of abnormal 
mitochondrial function. The trial design will be a 24-week randomized, DBPC cross-over study. 
We hypothesize that the supplement will result in a greater ability to normalize mitochondrial 
activity as compared to placebo. To test this hypothesis we will:   
a. Test mitochondrial activity at baseline and after the placebo and supplement arms of the 
study and determine if the nutritional supplement normalizes mitochondrial activity 
significantly more than placebo.  
b. Test redox metabolism at baseline and after the placebo and supplement arms of the study 
and determine if the nutritional supplement improves redox metabolism significantly more 
than placebo. 
c. To determine if specific biomarkers (Folate Receptor Autoantibody, Single Nucleotide 
Polymorphism) can predict response to treatment.  
Secondary Aim: To evaluate the effectiveness of a wide-spectrum metabolic supplement on core 
and associated ASD symptoms. We will determine is symptoms associated with ASD will be 
effected by a nutritional supplement which targets normalizing mitochondrial function. We will 
evaluate:    
a. Core ASD symptoms as measured by the Childhood Autism Rating Scale (CARS).  
b. Clinical Global Impression Severity and Improvement 
c.  Behavior, including repetitive behavior Children Yale Brown Obsessive Compulsive Scales 
Modified for ASD (CYBOCS-ASD), Irritability, Social Withdrawal, and  Hyperactivity subscales 
of the Aberrant Behavior Checklist (ABC), Parent-rated Anxiety Scale-ASD (PRAS-ASD), 
Caregiver Strain Questionnaire (CGSQ), Parent Targeted Problems and adaptive behavior 
(Vineland III). 
d. Somatic manifestations of pain, fatigue and gastrointestinal symptoms 
Tertiary Aim: To collect samples which will allow the evaluation of the effect of a wide-
spectrum metabolic supplement on changes in cellular regulation and to determine if 
specific gene abnormalities or cellular regulatory factors or autoantibodies results in 
abnormal mitochondrial function. In the future we wish to determine the changes in cellular 
regulation as a result of the treatment by examining several aspects of cellular regulatory 
elements. We also wish to determine if certain genetic or epigenetic changes are associated with 
mitochondrial dysfunction in ASD. The current funding does not allow these advanced analysis to 
be performed at this time but we will collect samples so these studies can be performed in the 
future. In the future we will examine: 
a. Gene expression by measuring RNA expression 
b. Cellular regulation by measuring microRNA expression 
c. Concentrations of key regulatory proteins 
d. Methylation of targeted genes 
e. Abnormal gene sequences (mutations, rare polymorphisms) 
f. Autoantibodies to Brain tissue and the folate receptor alpha  


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
7 
 
5. RESEARCH PLAN 
 
Trial Design  
50 children with ASD and signs and symptoms of abnormal mitochondrial activity will be randomized 
in 1:1 ratio to receive a wide-spectrum nutritional supplement that targets mitochondrial function or 
placebo for 12 weeks under DBPC conditions. Each participant will serve as their own control in the 
cross-over design. Using a phone screen, we will identify children (age > 2:6 and < 17:3) with a 
diagnosis of ASD and evidence of mitochondrial function which is significantly higher or lower than 
average (here forth referred to as mitochondrial dysfunction). Children with suspicion of mitochondrial 
dysfunction will be invited to participate in a screening visit to make sure they meet including and 
exclusion criteria. If the screening visit is longer than 6 weeks from the prescreening, we will repeat 
the baseline metabolic testing to ensure an accurate baseline mitochondrial measurement. If they 
meet criteria, the baseline visit will be completed within 2 week but we will attempt to complete it on 
the same day (Table 1). Upon successful completion of the baseline visit, the participant will enter 
Phase 1 of the trial and will be randomized to receive active treatment or placebo for 12 weeks under 
DBPC conditions. At the end of the 12 weeks (end of Phase 1), the participant will return for 
evaluations similar to the ones conducted at baseline. Upon successful completion of the post-Phase 
1 visit, the participant will start Phase 2 where they will receive active treatment or placebo, depending 
on what they received during Phase 1 – they will be crossed over to the opposite treatment regimen. 
Potential AEs, compliance and concomitant treatments will be monitored every four weeks. 
 
Inclusion criteria  
• 
Boys and girls ≥ 2 years 6 months of age to < 17 years 3 months of age; 
• 
Weight ≥ 15 kg and ≤ 100kg; 
• 
DSM-5 diagnosis of Autism Spectrum Disorder as established by formal clinical assessment which 
includes a gold-standard tool such as the Autism Diagnostic Observational Schedule. 
• 
Current Childhood Autism Rating Scale, 2nd Edition (CARS-2) score meeting criteria for Autism 
Spectrum Disorder (Standard Form, cutoff ≥ 30 for children < 13 years of age; ≥ 28 for children ≥ 
13 years of age; High Functioning Form, cutoff ≥ 28) 
• 
IQ at least 30 as measured by the Differential Abilities Scale (DAS)  
• 
Stable educational and therapy plan (one month) with no planned changes in the intensity of 
treatment for 12 weeks.  
• 
English spoken in the home and at least one parent is able to read, write and speak English.  
• 
Stable medication (no changes in past 6 weeks) and no planned changes for the study duration. 
• 
Signs and symptoms of mitochondrial dysfunction (See Figure Below) 
 
 
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
8 
 
 
 
Exclusion criteria 
• 
IQ below 30 as measured by the Differential Abilities Scale (DAS) 
• 
Presence of serious behavioral problems (tantrums, aggression, self-injury) for which another 
treatment is warranted.  
• 
Significant medical condition by history or by physical examination or lab tests that would be 
incompatible with the treatment.  
• 
Children taking anticonvulsant medication for seizures or active epilepsy.   
• 
Diagnosis of Mitochondrial Disease 
• 
Current use of SpectrumNeeds or QNeeds 
 
Procedures 
Randomization:  Randomization will be carried out within site using permuted blocks of 4 subjects 
without additional stratification to nutritional supplement or placebo in a 1:1 ratio. Statisticians will 
prepare the permuted blocks for each site and will be sent to the Investigational Pharmacist at each 
site. Investigators and study staff will be blind to the allocation sequence.  
 
Compensation Participants will be compensated $50 for each of the four physical visits.  
 
Measures: 
Parent Ratings 
1. Vineland Adaptive Behavior Social Subscale (VABS) III: The Vineland Adaptive Behavior Scale 
(VABS) III is a widely used standardized, well-validated assessment tool for children with 
developmental delays that measures functional abilities within several domains 66. It is particularly 
useful for children with intellectual disability which commonly co-occurs with ASD 67 and has valid 
measures of social impairments in children with ASD 68. The VABS relies on an informant (caretaker) 
to complete the survey about the child’s adaptive behavior – i.e., what the child actually does in the 
course of daily living. We will use the parent checklist version and examine the social domain. 
2. Aberrant Behavior Checklist (ABC): The ABC is a validated questionnaire that rates symptoms of 
hyperactivity, irritability, social withdrawal, and stereotypic behavior in individuals with developmental 
disabilities.69 It has convergent and divergent validity 70 and is used in multiple ASD clinical trials. 
3. Parent Rated Anxiety Scale-ASD (PRAS-ASD): Twenty five-item scale designed to measure 
anxiety in children with ASD. It has demonstrated reliability and validity (L. Scahill unpublished data). 
4. Caregiver Strain Questionnaire (CGSQ)-Short Form: This version of the CGSQ is a 7-item, parent 
self-report designed to measure the burdens associated with raising a child with special needs.   
5. Sensory Profile 2: This assessment provides information about sensory sensitivity and 
interpretation in children with developmental disorders. 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
9 
 
6. Parent Rated Autism Symptomatic Change Scale (PRASC): The PRASC is a 12-item questionnaire 
that asks parent to rate changes in symptoms from “much better” to “much worse” on a 7 point Likert 
scale from the beginning of the intervention. This instrument was developed by Frye and Rossignol 
(2010) and produces a reliable measure of changes The parent will be asked to complete this scale at 
each remote and onsite visit following randomization to monitor changes in ASD symptoms in the 
study. 
 
Table 1. Schedule of Measures 
 
 
Double blind Phase 
Measure 
Pre-Screening 
Screening 
Baseline 
*1 
wk 
*2 
wk 
*4 
wk 
*8 
wk 
12 
wk 
*16 
wk 
*20 
wk 
24 
wk 
Phase 1 
 
 
 
 
 
 
 
 
 
 
 
Phase 2 
 
 
 
 
 
 
 
 
 
 
 
Parent Ratings 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
Vineland III Caregiver 
Form [1] 
 
 
X 
 
 
 
 
X 
 
 
X 
ABC [2] 
X 
 
 
 
X 
 
 
X 
PRAS-ASD [3] 
 
 
X 
 
 
 
 
X 
 
 
X 
CGSQ-Short form [4] 
 
 
X 
 
 
 
 
X 
 
 
X 
Sensory Profile 2 [5] 
 
 
X 
 
 
 
 
 
 
 
 
PRASC [6] 
 
 
 
 
 
X 
X 
X 
X 
X 
X 
Laboratory 
Assessments 
 
 
 
 
 
 
 
 
 
 
 
Safety Laboratory [7] 
X 
 
 
 
 
 
 
X 
 
 
X 
Biomarker Laboratories 
[8] 
X 
 
 
 
 
 
 
X 
 
 
X 
Pregnancy (B-HCG)  
X 
 
 
 
 
 
 
 
 
 
X 
Vital Signs 
X 
 
 
 
 
 
 
 
 
 
 
Height and Weight 
X 
 
X 
 
 
 
 
X 
 
 
X 
Head Circumference 
X 
 
 
 
 
 
 
 
 
 
 
Medical Assessments  
 
 
 
 
 
 
 
 
 
 
 
Medical/Psych History [9] 
X 
 
 
 
 
 
 
 
 
 
Physical Exam  
 
X 
 
 
 
 
 
 
 
 
 
Adverse Effects Review  
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Concomitant Treatments 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Suicidality Assessment 
[10] 
 
X 
X 
 
 
X 
X 
X 
X 
X 
X 
Compliance 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
Developmental 
Assessments 
 
 
 
 
 
 
 
 
 
 
 
CARS [11] 
 
X 
 
 
 
 
 
X 
 
 
X 
DAS [12] 
 
X 
 
 
 
 
 
X 
 
 
X 
OACIS [13] 
 
 
X 
 
 
 
 
X 
 
 
X 
CGI-Severity [14] 
 
X 
 
 
 
 
X 
 
 
X 
CGI-S-ASD  
 
 
X 
 
 
 
 
X 
 
 
X 
CGI-Improvement [15] 
 
 
 
 
 
 
 
X 
 
 
X 
CGI-I_ASD  
 
 
 
 
 
 
 
X 
 
 
X 
Parent Target Problems 
[16] 
 
X 
 
 
 
X 
 
 
X 
CYBOCS-ASD [17]  
 
 
X 
 
 
 
 
X 
 
 
X 
Other Assessments 
 
 
 
 
 
 
 
 
 
 
 
Eye tracking task [18] 
 
 
X 
 
 
 
 
X 
 
 
X 
 
 
 
 
 
 
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
10 
 
Laboratory Assessments 
7. Safety laboratories: Safety labs will be measured at the screening visit. These will include a 
complete blood count and a comprehensive metabolic panel.  
8. Biomarker: Most biomarkers will be collected at baseline and after each treatment arm to determine 
if the treatment results in a change in these biomarkers. Baseline biomarkers will help understand 
whether treatment response can be predicted from biomarker tests. 
a. Buccal swabs will be collected to assess mitochondrial function.  
b. Plasma, Peripheral Blood Mononuclear Cells (PBMCs), DNA, and RNA will be collected and stored 
locally for future analysis, such as oxidative and methylation metabolism, bioenergetics, immune 
biomarkers, gene expression, epigenetics, and gene sequencing. 3billion Laboratory in Seoul, 
South Korea will perform genetic sequencing to look for causative mutations if good quality DNA is 
obtained and can be isolated. Samples will be stored until they are used up or destroyed, when 
possible. The PI, Dr. Richard Frye, will retain control over the blood sample unless it is requested 
that the sample be destroyed and then it will not be used for any future studies. If the participant 
wishes to have the sample removed from storage and destroyed at any time, they may contact a 
study coordinator or PI. IRB permission will be obtained prior to sharing samples for future studies. 
These samples will be de-identified and no protected health information will be shared with 
collaborators. If samples are shared, a log of those who withdraw samples will be documented 
thoroughly.  
c. Deciduous teeth and hair samples will also be collected and sent to Dr Arora’s laboratory. Recent 
research suggests prenatal nutritional metal concentrations measured in deciduous teeth correlate 
with long-term language development and may predict treatment response and other recent studies 
support the measurement of unique metabolic markers using hair samples. 
d. Folate receptor α autoantibody (FRAA) titers will be sent to the laboratory of Dr. Edward Quadros, 
Ph.D. at the State University of New York, Downstate (Brooklyn, NY) or Iliad Neurosciences. We 
have demonstrated in one study that FRAA are related to methyl-B12 metabolism.71  
e. Saliva will be collected from the participant and biological parents to investigate whether single 
nucleotide polymorphism (SNPs). Parental SNPs are important to understanding whether the 
SNPs in the children are significant or not – for example, if parents have the same SNP profile as 
the child, the SNP is unlikely to have a significant effect of pathophysiology.  
f. Autoantibodies to neuronal tissue may be assessed by the Laboratory of Dr. Madeleine 
Cunningham at the University of Oklahoma Health Sciences Center in Oklahoma City, OK if 
enough sample is available for the assay. 
 
Medical Assessments 
9. Medical and Psych History: This structured questionnaire form assists with the elicitation and 
assessment of health complaints, appetite, sleep, activity level and general health at baseline. The 
companion measure, the Adverse Effects Review, is used in follow up visits to monitor and document 
new events during the trial. 
10. Suicidality Assessment: Although we do not anticipate any symptoms consistent with suicidality, 
an assessment of suicidality is required in FDA trials involving psychiatric disorders including ASD 
and is a standard assessment in FDA regulated ASD trials. An assessment similar to other studies 
examining ASD children will be used. The Columbia-Suicide Severity Rating Scale (C-SSRS) fulfills 
this requirement.  
 
Developmental Assessments 
11. Childhood Autism Rating Scale (CARS): This 15-item scale can be used to both diagnose autism 
and ASD, and to assess the overall severity of symptoms. 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
11 
 
12. Differential Abilities Scale (DAS): This scale is a scale that assesses cognitive abilities and early 
learning and has been validated against other cognitive ability scales (Wechsler, Bayley, etc.). This 
scale is designed to identify clinical risk and disability. 
13. Ohio Autism Clinical Impression Scale (OACIS): A clinical evaluation by the study physician of 
initial severity of 10 ASD-related symptoms at baseline on a 7-point scale (Part 1), and a clinical 
evaluation of change of those symptoms using a different 7-point scale (very much improved to very 
much worse).  The 10 items include social interaction, aberrant/abnormal behavior, repetitive/ritualistic 
behavior, verbal communication, non-verbal communication, hyperactivity/inattention, anxiety/fears, 
sensory sensitivities, restricted/narrow interests, and autism. 
14. Clinical Global Impression - Severity (CGI-S): This is a 7-item scale ranging from a score of 1 for 
“Normal” to 7 for “Extreme” 72. All aspects of the child’s condition need to be integrated to assign the 
CGI-S score. Only participants of CGI-S of 4 (“moderate”) or greater severity will be included in the 
trial. By definition, a child with ASD and language impairment (which is established at screening) has 
moderate severity ASD. Thus, the CGI-S will be completed at baseline since it requires all available 
information about the child. CGI-S will be rated by an assessor who is blind to treatment assignment. 
Both the standard CGI-S and the newly developed CGI-S-ASD will be completed, with the CGI-S 
completed before the CGI-S-ASD. 
15. Clinical Global Impression for Improvement (CGI-I): The CGI-I is a 7-point measure of overall 
symptomatic change compared to baseline 72 that will be used as a key primary outcome measure. 
Scores range from 1 (Very Much Improved) through 4 (Unchanged) to 7 (Very Much Worse). All aspects 
of the child’s condition need to be integrated to assign the CGI-I score. The CGI-I will be rated by an 
assessor who is blind to treatment assignment, AEs, and medication dose. Both the standard CGI-I and 
the newly developed CGI-I-ASD will be completed, with the CGI-I completed before the CGI-I-ASD. 
Ratings of Much Improved or Very Much Improved on the CGI-I will be used to define positive response. 
16. Parent Target Problems (PTP): At baseline, parents will be asked to nominate the two most 
important ASD problems for the child. Through brief discussion, the frequency (for episodic behaviors 
such as insists on daily routines) or constancy (for problems such as language delay or rigidity 
reflecting more enduring patterns), intensity and impact of the behavior on the family are established. 
Responses from this systematic inquiry are documented in a brief narrative. The narrative will be 
reviewed monthly and a new narrative documented. This review will assist with the scoring of the CGI-
I in real time. 73 At the end of the trial, the narratives will be reviewed by a panel blind to treatment 
assignment. This method has been shown to be reliable and valid in placebo-controlled studies.73,74 
17. Children’s Yale-Brown Obsessive-Compulsive Scales-ASD (CYBOCS-ASD): The CYBOCS-ASD 
is a modified version of the CYBOCS developed for children with Obsessive-Compulsive Disorder.75 
The modified version is a semi-structured clinician-rated scale designed to rate the current severity of 
repetitive behavior in children with ASD (i.e., Time Spent, Interference, Distress, Resistance, and 
Control).  Each item is scored from 0 (least symptomatic) to 4 (most symptomatic), yielding a Total 
score from 0 to 20. It has established reliability and validity 76 and is sensitive to change.77   
 
Other Assessments 
18. Eye Tracking Task: This 15-item scale can be used to both diagnose autism and ASD, and to 
assess the overall severity of symptoms. The foremost criterion for the diagnosis of autism spectrum 
disorder (ASD) is a pervasive deficit in social communication and interaction, including abnormal 
speech, impaired reciprocity in relationships and conversation, and poor use and integration of 
nonverbal communication behaviors. A key behavior indicative of impaired social function in ASD is 
inappropriate or absent eye contact. For example, when gazing at faces, individuals with autism tend 
to direct gaze toward the mouth rather than the eyes. Changes in social communication are typically 
measured using multiple clinical assessments and caregiver-reported questionnaires normed for the 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
12 
 
ASD population. We wish strengthen measures of abnormal attention to social stimuli. We believe that 
objective changes in attentional gaze will be reflected in clinical presentation and overall clinician and 
caregiver reports of symptom severity. We may use the Tobii eye tracking software and Noldus Face 
Reader software to collect data about the location and duration of eye gaze while participants watch 
an age-appropriate video. The video will include both social and nonsocial stimuli. In each age-
appropriate video, we will present a variety of simulated social interactions as well as nonsocial 
behaviors. We will also present speech sounds and non-speech sounds, as well as background 
distractors. Each of these elements will be presented across the four quadrants of the screen. We will 
measure the percentage of time the participant spends gazing at social stimuli and nonsocial stimuli, 
as well as time spent away from the screen. We will also analyze the duration of fixation for each shift 
in gaze. This will be performed if equipment is available and operational.  
 
Investigational Product 
Table 2. Daily Serving Size 
  
6.6g 
8.8g 
11g 
13.2 g  
Tolerable 
Upper 
Intake 
Levels 
Patient Body Weight                         Units 
 
15kg-
20kg 
21kg-
40kg 
41kg 
- 
60kg 
61kg 
- 
100kg  
Age 4-8 
years 
NeuroNeeds 
 
 
 
 
 
 
Vitamin C (ascorbic acid) 
Mg 
450 
600 
750 
900 
ND 
Vitamin D (cholecalciferol) 
Mcg 
11.25 
15 
18.75 
22.5 
75 
Vitamin E (d-α-tocopheryl acetate 
and mixed tocopherols) 
Mg 
37.5 
50 
62.5 
75 
300 
Thiamin (as thiamin HCl) 
Mg 
18.75 
25 
31.25 
37.5 
ND 
Riboflavin  
Mg 
18.75 
25 
31.25 
37.5 
ND 
Niacinamide 
Mg 
18.75 
25 
31.25 
37.5 
151 
Vitamin B6 (50% as pyridoxine HCl 
and 50% as pyridoxal-5-phosphate) 
Mg 
15 
20 
25 
30 
40 
    (6S)-5-methyltetrahydrofolic acid 
mcg  
1250 
1667 
2084 
2500 
ND 
    Calcium folinate 
mcg 
1250 
1667 
2084 
2500 
4002 
Vitamin B12 (as methylcobalamin) 
Mcg 
375 
500 
625 
750 
ND 
Biotin 
Mcg 
750 
1000 
1250 
1500 
ND 
Pantothenic acid (as D-Ca 
pantothenate) 
Mg 
225 
300 
375 
450 
ND 
Calcium 
Mg 
45 
60 
75 
90 
2,500 
Iodine (as K iodide) 
Mcg 
75 
100 
125 
150 
300 
Magnesium (Mg citrate and Mg 
malate) 
Mg 
150 
200 
250 
300 
110 
Zinc (Zn gluconate) 
Mg 
9 
12 
15 
18 
12 
Selenium (as selenomethionine) 
Mcg 
37.5 
50 
62.5 
75 
150 
Manganese (as manganese amino 
acid chelate) 
Mg 
0.75 
1 
1.25 
1.5 
3 
Chromium (Cr picolinate) 
mcg 
37.5 
50 
62.5 
75 
ND 
Molybdenum (Na molybdate) 
mcg 
75 
100 
125 
150 
600 
Potassium (K citrate) 
mg 
37.5 
50 
62.5 
75 
 
Creatine monohydrate 
mg 
937.5 
1250 
1562.5 
1875 
 
L-Arginine 
mg 
750 
1000 
1250 
1500 
 
Carnitine Blend (acetyl L-carnitine 
HCl and L-carnitine) 
mg 
375 
500 
625 
750 
 
Alpha ketoglutaric acid 
mg 
225 
300 
375 
450 
 
Ubiquinone 
mg 
187.5 
250 
312.5 
375 
 
Choline bitartrate 
mg 
112.5 
150 
187.5 
225 
1000 
Inositol 
mg 
75 
100 
125 
150 
 
Alpha lipoic acid 
mg 
75 
100 
125 
150 
 
D-Beta, D-Gamma, D-Delta 
tocopherols 
mg 
3.3 
4.4 
5.5 
6.6 
300 
QNeeds 
 
 
 
 
 
Ubiquinol (QNeeds)4 
mg 
200 
300 
400 
400 
 
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
13 
 
The nutritional supplement used will be a combination of SpectrumNeeds and QNeeds, both 
produced by NeuroNeeds (Old Lyme, CT; See Table 1). Weight based dosing will be use. The daily 
serving size will divided into two daily doses in the form of a powder which can be mixed into liquid of 
food.  Together, there are 34 different dietary supplements in the products. Thirty-two of the 34 
supplements do not exceed the Tolerable Upper Intake Limits set by the United States Institute of 
Medicine. The exceptions are:  
1. The niacin dosing is about the upper limit. Niacin in the form of nicotinic acid is well known for 
causing benign flushing. Niacinamide (also known as nicotinamide) is the form of niacin in 
SpectrumNeeds, and this form does not cause flushing. 
2. Folate exceeds the upper limit. It should be noted that half of the total folate provided is in the form 
of Quatrefolic™ (6S)-5-Methyltetrahydrofolic acid Glucosamine salt (55% FOLATE), which is 
microbial sourced, not synthetic. There is no known upper tolerable limit for non-synthetic folate, 
only for synthetic folate. The other half is in the form of calcium folinate. Dosage of folate used in 
this supplement are about one-half that commonly recommended during pregnancy to women with 
a prior child with a neural tube defect who may get pregnant. Dosages of folinate are one half to 
one-twenty-fifth the dosing commonly recommended in children with ASD. These dosages are 
known to be generally well tolerated in ASD children. 
 
Except for ubiquinol, all of these nutrients are provided in a powder form in SpectrumNeeds. The 
physical stability and bioavailability of ubiquinol is insufficient in powder form, and thus ubiquinol is 
provided separately in QNeeds gel capsules. These capsules can be swallowed whole, or cut with 
scissors and the contents squeezed out and added to SpectrumNeeds just before ingestion. 
 
Placebos will have the same look, feel and taste as the active treatments. For example, the 
SpectrumNeeds is a silica powder flavored by monk fruit extract, so the inert silica powder with the 
same favoring will be used as a control. Similarly, in QNeeds, the ubiquinol is carried by purified water 
and glycerin colored by caramel liquid, so for the placebo the same carrier and color will be used in 
the gel capsule without ubiquinol. 
 
Ongoing Examiner Reliability and Fidelity Assessments: A psychologist will train assessors at each site 
on the administration of the confrontational assessment. The psychologist will review the first three 
administrations of the language test to confirm reliable administration and conduct quarterly conference 
calls to review cases and to promote a common approach to conducting assessments across sites.  
 
Data Management: Study data will be collected and managed using the REDCap software specifically 
developed for research and clinical trial data collection and management. REDCap is a secure web 
application and fully compliant with privacy guidelines and communication security as defined in the 
HIPAA, Federal Information Security Management Act (FISMA), and the U.S. Food and Drug CFR for 
electronic records. 
Food and Drug Administration (FDA) Approval  
The FDA has approved our Investigational New Drug (IND) application.  
 
6. DATA MANGEMENT AND STATISTICS 
 
Storage and Security  
Study data will be managed using a 21 CFR Part 11 compliant web-based data entry system. Thus, 
data collection templates will be modified accordingly for data entry.  
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
14 
 
Statistical Analysis 
All data analysis for the primary study aims will be conducted by the principal investigator.  
The primary analysis will be intent-to-treat, defined as all randomized subjects. Exploratory “per 
protocol” analysis will be conducted on participants who adhered to protocol. Analyses will be 
performed with R 3.02 (http://www.r-project.org) or SAS 9.3 (SAS Institute, Cary, NC).  
 
Sample Size Justification and Power Analysis: Assuming a medium effect size of 0.5, the study 
provides an 80% power assuming a 0.01 alpha (allowing correction for multiple primary end-points)..   
 
Primary Hypothesis:  After 12 weeks of taking the nutritional supplement will improve mitochondrial 
function more than taking the placebo for 12 weeks. To test this hypothesis, we will use a mixed 
model using baseline and all post-randomization measures of mitochondrial activity. To deal with 
missing post-treatment data, the model will be conditioned on all baseline values.72 This conditional 
joint response model is more tolerant of missing data than ANCOVA and is less biased than carrying 
baseline data forward to endpoint.   
 
Safety analyses:  Adverse events (AE) will be systematically monitored using a 34-item form. It 
includes specific queries about major body systems, activity level, sleep, appetite, and general health. 
New events, whether presumed related to treatment or not, are classified as AEs and rated as Mild, 
Moderate or Severe (see details below). The occurrence of an AE will be counted once at the highest 
level of severity (e.g., a report of mild nausea followed by a report of moderate nausea in the same 
child would counted as an occurrence of moderate nausea). The frequency of AEs by severity will be 
evaluated using a Fisher’s exact test. We will pursue exploratory analyses such as survival analyses 
using time to AE as the response variable.  
 
Validation of the CGI-ASD: In addition to the standard CGI-S and CGI-I we will use new a structured 
CGI rating scale called the CGI-S-ASD and CGI-I-ASD respectively. We will compare the performance 
of the standard and new CGI rating scales. Since the CGI-ASD provide guidance for completing the 
ratings we will have the rater complete the standard scale before the new scale. The correspondence 
between each scale and other outcome measures will be studied as well as the performance of the 
scales as outcome measures.  
 
 
Extra Sample Collection: As an optional portion to the research, deciduous teeth and hair samples can 
also be collected and sent to Dr Arora’s laboratory. Recent research suggests prenatal nutritional metal 
concentrations measured in deciduous teeth correlate with long-term language development and may 
predict response to various treatments and other recent studies support the measurement of unique 
metabolic markers using hair samples. 
 
7. HUMAN SUBJECTS CONCERNS 
 
Sources of Materials 
The sources of the research material will be data collected in the course of the clinical trial to 
determine the efficacy and also to ensure the safety of study participants. The principal types of data 
will be clinical rating scales and standardized tests (e.g., IQ test and the CELF-4). Routine laboratory 
measures (blood chemistry, blood cell counts) and biomarkers will be obtained.  
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
15 
 
Future Studies 
Although the long-term use of the SpectrumNeeds and QNeeds multivitamin is believed to be safe, 
long-term tolerability and therapeutic effect has not been evaluated. Furthermore, the optimal length of 
treatment is not known. Further studies will identify the optimal treatment for this therapy and long-term 
safety.  
 
The biomarkers to be collected for future research include DNA, and RNA. These samples will be 
collected and sent to Dr Frye’s laboratory for storage for future analysis such as gene expression, 
epigenetics, gene sequencing, proteomics or metabolomics. 
 
Potential Risks  
The risks associated with these procedures include exposure to the study supplement or placebo, and 
the risk associated with venipuncture.  The blood draws poses a possible risk of bruising at the site of 
the draw, light-headedness and fainting along with a remote chance of infection.  
 
Although highly unlikely, there are potential privacy risks associated with participating in such a study.  
There is minimal risk associated with the completion of rating scales and assessment measures.  
Blood draws are common procedures and will be performed by a certified phlebotomist. We will offer 
a numbing cream prior to the blood drawing procedure. We will systematically review adverse at 
follow up phone calls and clinic visits (see below). Parents will be given phone numbers of the 
principal investigator and study physician – who will be available after hours. The PI and study 
physician have access to the pharmacy after hours – if it is necessary to break the blind. 
 
The nutritional supplement and supplements similar to it have been on the market and used widely in 
the autism community. There have been few reported side effects of this supplement. They include 
nausea and hyperactivity, both of which resolve on their own. One similar supplement produced by Dr. 
Adams at Arizona State University has been studied in three clinical trials without reported significant 
adverse effects. In addition, the majority of the ingredients in the nutritional supplement are well within 
the proposed upper tolerable limits recommended by the institute of medicine and the two vitamins 
that exceed these limits are known to be well tolerated at doses well above the recommend upper 
limits. Increased consumption of B vitamins may also temporarily change urine color.  
 
Confidentiality will be maintained throughout the project period. With the exception of tests that 
require date of birth, research records will not include names or contact information. Subjects will be 
identified only by their code numbers. A file with personal health information (e.g., names and 
addresses) will be kept separately in a locked file cabinet.   
 
Recruitment   
The research team will use targeted recruitment, such as posting to the local website, contacting 
clinical populations, using social media and contacting local organizations, physicians and schools. 
All recruitment materials such as flyers, letters and social media posts will be IRB approved. There 
must be a suspicion of mitochondrial dysfunction for the participants to qualify for the study.  
  
Compensation  
Subjects will be compensated $50 for the following visits: screening, baseline, end of Phase 1 and 
end of Phase 2.  
 
Consent Process  


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
16 
 
Interested families will be screened on the telephone by the study coordinator. Presumably eligible 
and willing families will be invited to the site for a formal screening visit.  Parental consent will be 
obtained at the screening visit prior to the collection of any study data. The consent procedure will 
include the following: (a) provide the consent forms to parents and guardians together with a verbal 
summary of the contents of all paragraphs, and (b) a description of alternatives to participating in the 
study (not to take part, use of other treatments, and obtaining similar treatment through other clinical 
services). Parents (guardians) will also be told that declining participation will not prejudice their right 
to pursue other treatments at the local site or any of its affiliates.  
 
Parental consent will be obtained by trained investigators as documented on each sites delegation of 
authority log. Children whose parents indicate notable barriers to meeting the demands of the study or 
whose parents lack capacity to understand the purpose and expectations of the study will not be 
randomized in the study. These judgments will be made by the investigator conducting the consent 
discussion in consultation with the research team as needed.  Parents will be given the opportunity to 
ask questions. Once all questions have been answered, parent(s) will be asked to sign and date the 
consent form. Parents will be provided with a copy of the singed consent form.  
 
Child Assent: The children in this study will be diagnosed with ASD. Most will also be cognitively 
delayed. The site investigator or designated Co-investigator conducting the consent procedure will 
evaluate the child’s capacity to provide written or verbal assent. The investigator’s judgment will be 
documented on the assent form.    
 
Consent for Video Recording Consent for video recording for research purposes will be obtained from 
parents. If parents are willing, video recordings will be used for verifying behavioral scoring but 
additional observers, monitoring of the testing for ensuring high quality evaluations and training 
evaluators. Parents can indicate agreement or disagreement with this additional use of the videos 
directly on the video consent form. Agreement to use the videos for these additional training purposes 
is not required for study participation.  
 
Parental Consent for Saliva Biological parents may choose to provide saliva samples. This is not 
required for child participation in the study. Parents will be asked to accept or decline by initialing 
under the "Making Your Choice" option in the future and genetic research sections. There will also be 
a separate consent page for each biological parent at the end of the consent form. If one of the 
parents is unable to consent to providing saliva at the time of initial consent, they may return to the 
clinic and be consented by the PI at a later date. This will be documented in the subject record, and 
the family will be provided a copy of the updated consent form. 
  
Study Withdrawal Parents are free to withdraw from the study at any time. This will be clearly stated 
during the consent process. Subjects may need to discontinue participation in the study before 
completion for various reasons (e.g., family move, withdrawal of consent, adverse events, need for a 
different treatment). Parents who indicate intention to withdraw the child from the study will be offered 
to have a case conference with the study team to discuss the matter.  If the discussion indicates that 
the child needs another treatment, we will assist the family to locate that treatment. Parents who drop 
out of treatment will be invited to return for scheduled assessments or an early termination visit. Data 
collected from children who drop out of treatment, but return for assessments, will be analyzed in their 
originally randomized group.  
 
CONFIDENTIALITY 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
17 
 
Case report forms (CRFs) contain the subject’s unique ID number – but no identifying information. 
The RedCap Clinical Trial Management System is a secure, password protected data base. The 
folders containing CRFs data will be kept in locked files and access to these files is only granted to 
members of the research team. Standardized tests (e.g. IQ and language tests) record will not record 
the child’s date of birth but will use age calculate normative data. No other personal health information 
will be documented on research forms. Contact information for the family is kept in a separate file that 
is in a locked cabinet.  
 
Certain measures will be recorded on video. The digital files are stored on a secure, password-
protected site. Recordings may also be stored electronically on an encrypted external hard drive that 
is kept in a locked closet or cabinet. The digital files will be marked with a Subject ID instead of names 
of children or their parents. It is possible that names may be mentioned on the recordings. Video 
recordings from this research study will be retained for up to 5 years after the study is over. 
Recordings may be used for other purposes (e.g. presentations at professional meetings) if separate 
consent is obtained from the parent (or guardian).  
 
Access to study records (case report forms) will be restricted to study staff.  Others that may review 
research records include study monitors, representatives from Food and Drug Administration (FDA). 
We may submit de-identified study data to the National Database for Autism Research (NDAR).  
 
Potential Benefits of the Proposed Research to the Subjects and Others 
Participants may directly benefit by the receipt of the supplement.  Overall, the risks to the subjects in 
this research are similar to those that exist in standard clinical practice and the proposed research 
project offers considerable promise in improving mitochondrial function in children with ASD. Indeed, 
this research may lead to a new treatment option for children with ASD. Thus, the potential benefits of 
the research outweigh the risks involved.  
 
Importance of the Knowledge To Be Gained Children with ASD are impaired and, untreated, their 
long-term prognosis is guarded. Although current treatments are helpful in some cases, significant 
gaps in our treatment options remain. There are no large, controlled clinical trials of nutritional 
supplements in children with ASD and no medication treatments for core features of ASD or language 
delay. This study may provide new information about nutritional supplements in children with ASD and 
may identify subgroups of affected children who could benefit from a nutritional supplement.  
 
Data Safety and Monitoring Plan The medical monitor will review adverse effects every three 
months. If potential safety concerns are identified, the medical monitor will review the risk-benefit ratio 
with the unblended statistician. If treatment-emergent safety concerns alter the risk-benefit ratio, the 
medical monitor will contact the PI for discussion of potential AEs. 
 
Monitoring for Adverse Events (AEs):   
At each visit, including baseline and interim contacts, the treating clinician will systematically review 
for AEs and concomitant medications using an AE Review form developed for this study. We have 
used versions of this form, in several multi-site clinical trials in children with ASD which contains a 
general inquiry including several questions about daily activities (e.g., sleep, appetite, energy level, 
bowel and bladder functions). The general inquiry includes an open-ended question about any 
problems or complaints, as well as questions regarding the need for other medications and doctor or 
health care encounters since the last study visit. The last section includes specific questions about 
daily activities. All new AEs will be documented on the AE log. The status of previously-reported AEs 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
18 
 
will be monitored as well.  
 
When the treating clinician elicits an AE, it will be documented regardless of suspected relationship to 
the supplement. The AE log requires the treating clinician to label the AE and to document the 
severity, onset, course, outcome, and attribution (study drug related or not).    
 
Attribution is classified as follows:  
•  Definite: AE is clearly related to the study drug.  
•  Probable: AE is likely to be related to the study drug.  
•  Possible: AE may be related to the study drug.  
•  Unlikely: AE is doubtfully related to the study drug.  
•  Unrelated: AE is clearly not related to the study drug.  
 
 Severity of AEs is classified as follows:  
• mild= present, but no intervention required;  
• moderate = present, may be bothersome or may require intervention;  
• severe = present, bothersome, requires intervention and may produce sequelae.  
  
Decisions on the appropriate care of the subject will be made by the treating clinician (who will remain 
blind to treatment assignment), in collaboration with the family’s primary care provider whenever 
possible.  Abnormal findings on laboratory tests that exceed established alarm values will be 
investigated to determine the nature of the adverse events.  
   
Routine reporting of AEs Adverse events will be documented on the Adverse Event Log as described 
above.  All AE data will be captured in the electronic database. Investigators will provide this 
information to local IRBs at the time of annual renewal. The investigators will decide if modifications to 
the protocol or consent form are required based on ongoing AEs. 
 
AEs resulting in treatment discontinuation:  Adverse events that are the primary reason for study 
discontinuation will be tracked separately. High rates of such events in the active supplement group 
compared to the placebo group will prompt evaluation of potential significant safety concerns in the 
trial. Rates of AEs will also be monitored during the open-label extension trial. 
 
Serious adverse events:  A serious adverse event (SAE) is defined as an event that poses a threat to 
the participant’s life or functioning. We note that “severe” is not necessarily equivalent to “serious.” 
Thus, a severe rash is not a serious adverse event, whereas a heart attack of any severity is likely to 
be a serious adverse event. A serious adverse event is defined as any event that entails one of the 
following:  
• Death;  
• Threat to the individual’s life;  
• Inpatient hospitalization or prolongation of existing hospitalization;  
• Persistent or significant disability or incapacity;  
• Intentional drug overdose;  
• Any other significant event that jeopardizes the participant.  
 
 
 


Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
19 
 
Assessment of Serious Adverse Events: The SAE will be classified as: a) serious AND anticipated; b) 
serious AND anticipated, BUT occurring with a greater frequency than expected; c) serious AND 
unanticipated.  SAEs that are unanticipated or anticipated SAEs that occur at a greater 
frequency than expected and deemed at least possibly related to the study treatment will be 
reported with seven days.  The report will be sent to study IRBs. The report will be sent to the FDA 
within 14 days.  SAEs that do not meet either of these criteria will be included in annual reports to the 
IRBs. 
 
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Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
20 
 
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Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
21 
 
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Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
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Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
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Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
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Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
24 
 
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Full Study Title: Effect of a Wide Spectrum Nutritional Supplement on Mitochondrial Function in Children with Autism 
Spectrum Disorder (ASD) 
Principal Investigator: Richard Frye; Co-Investigators: Richard Boles 
Sponsor: Dr. Richard Frye; IND 142751; ClinicalTrials.gov ID: NCT03835117; Protocol Version 02/11/2022 
Funding Source: Turnabout for Autism (Private Foundation), Pleasanton CA.  
 
25 
 
modified Children’s Yale-Brown Obsessive Compulsive Scale to detect change: results 
from two multi-site trials. , Autism: The International Journal of Research and Practice, Vol. 
19(1) 38–52, DOI: 10.1177.