Sulforaphane Treatment of Children With Autism Spectrum Disorder (ASD)

Background: ASD is a diverse disorder starting in early childhood and characterized by social
communication impairment as well as restricted interests and repetitive behaviors. It affects
1:68 children and is an enormous medical and economic problem for which there is no
established, mechanism-based treatment. Sulforaphane is an isothiocyanate derived from
broccoli, and has potent activity in transcriptionally up-regulating genes that control
mechanisms whereby aerobic cells protect themselves against oxidative stress, mitochondrial
dysfunction, and inflammation.

Hypothesis: Based on the observation that fever is frequently associated with behavioral
improvements in children with ASD, it is hypothesized that sulforaphane might lead to
functional improvements in ASD because it can up-regulate cell-protective responses, such as
heat shock proteins and related mechanisms that are also up-regulated during fever. These
mechanisms are central to multiple cellular processes in the central nervous system,
including synaptic transmission, and may improve long-range cerebral cortical connectivity,
which is depressed in ASD.

Specific aims: The 3 specific aims in this study are: 1) To determine if there are measurable
effects on social responsiveness and problem behaviors during treatment of children with
sulforaphane; 2) To determine if treatment with sulforaphane in children is safe and well
tolerated; and 3) To elucidate cellular biomarkers that respond to treatment with
sulforaphane.

Design: This is a randomized, double blind, single-arm crossover phase 1/2 clinical trial of
orally administered sulforaphane (as broccoli seed powder) in 50 boys and girls (3-12 years)
with ASD in 3 phases over 36 weeks.

In Phase 1, 25 children will receive active drug and 25 will receive placebo for 15 weeks; in
Phase 2, all children will receive sulforaphane from 15-30 weeks; in Phase 3, children will
receive no treatment for 6 weeks. Study visits will take place at screening, 7, 15, 22, 30
and 36 weeks, when the Ohio Autism Clinical Clinical Impressions Scale - Severity and
Improvement (OACIS-S and OACIS-I), Aberrant Behavior Checklist (ABC) and Social
Responsiveness Scale (SRS) will be recorded.

The children will be monitored with physical examinations and for toxicity with clinical
laboratory studies and examine possible biomarkers: Nrf2, oxidative stress and mitochondrial
function, mTOR pathway and cytokine expression.

In addition, prior to the main clinical trial, a pilot study will be performed in 10 children
with ASD, 6-12 years of age, who will receive sulforaphane, 2.2 micromoles/kg daily for 14
days. Blood and urine samples before and at the end of treatment will be collected, in order
to measure several parameters that are likely to demonstrate expected effects of
sulforaphane, to standardize the assays and procedures, and to determine the most effective
measures.

Inclusion Criteria:

- Autism Spectrum Disorder (ASD) diagnosis of moderate or greater severity

- Age 3 through 12 years inclusive

Exclusion Criteria:

- Absence of a parent or legal guardian and consent

- Inability to speak/understand English language

- Seizure within 1 year of screening: This exclusion is based on the theoretical concern
that cellular activation by sulforaphane might exacerbate seizures in patients with
known seizure disorders. As noted in our previous trial of sulforaphane in young adult
males, a seizure occurred in each of 2 participants: one during treatment (in a
participant with a previously undisclosed seizure), the other 3 weeks after
discontinuing sulforaphane.

- Impaired renal function (serum creatinine > 1.2 mg/dl), impaired hepatic function
(SGOT/SGPT> 2x upper limit of normal), impaired thyroid function (Thyroid Stimulating
Hormone (TSH) outside normal limits): This exclusion is based on a theoretical
possibility of activation of underlying cellular metabolic abnormalities by
sulforaphane. Current infection or treatment with antibiotics: this exclusion is to
avoid complications of inter-current illness that may occur due to the clinical trial
or obscure possible effects of sulforaphane.

- Medications that may modify the course or testing of ASD parameters (e.g.,
prednisone): This exclusion is necessary in order not to interfere with or complicate
effects of sulforaphane.

- Chronic medical disorder (e.g., cardiovascular disease, stroke or diabetes) or major
surgery within 3 months prior to enrollment: Serious medical illness in the child may
be complicated by the clinical trial and make it difficult to discern a change in ASD
associated with treatment.

- Less than 3 years or more than 13 years of age: this age range was selected to cover
the ages from usual diagnosis of ASD up to adolescence.

- A diagnosis of autism spectrum disorder of mild severity (for example, earlier
categories of Asperger disorder, Pervasive Developmental Disorder - Not Otherwise
Specified (PDD-NOS)), according to Autism Diagnostic Observation Schedule (ADOS)
criteria.

- Prisoners

- Pregnant women