Sulforaphane-rich Broccoli Sprout Extract for Autism

Behavioral improvements occur transiently during febrile illnesses in autism, and include
decreased repetitive behaviors and improved speech. These changes have been recorded in 38%
of autistic children in a clinical survey and 83% in an observational study, respectively.
The cellular basis for this "fever effect" is unknown but is likely to involve heat shock
proteins (HSP) and cellular stress responses (CSR) that lead to changes in synaptic function
and network connectivity.

Sulforaphane (1-isothiocyanato-4R- (methylsulfinyl)butane) is an isothiocyanate that is
delivered by lyophilized extracts of 3-day-old broccoli sprouts. Broccoli sprouts are widely
consumed as a food item all over the world by very large numbers of individuals, without any
reports of adverse effects. Our preliminary work in vitro shows that sulforaphane stimulates
HSP and mitochondrial biogenesis in several genetic disorders.

This study of sulforaphane-rich Broccoli Sprout Extract in autism is a randomized,
double-blinded, placebo-controlled, phase II single site trial, designed to ensure safety and
obtain efficacy data, with a focus on changes in social responsiveness, a core feature of
autism. Its hybrid design, incorporating double masking, placebo control, and randomization,
enhances the robustness of early outcome data. The study duration will be 2 years.
Recruitment of subjects will be 50% by 8 months and complete by 14 months. All subjects in
the study will be followed for 22 weeks. Treatment (18 weeks) will be started as patients are
entered into the study and receive baseline testing. All treatment will be completed by 20
months and data analysis and presentation of results by 24 months.

Forty-five male adolescents (13-18 years) and adults (19-30 years) with autism will be
randomly assigned to receive either sulforaphane-rich Broccoli Sprout Extract (n = 30) or
placebo (n = 15). The 2:1 randomization schedule will be produced by the study statistician
using permuted random blocks and stratification by history of positive behavioral effects of
fever. Treatment assignments will be performed by the research pharmacy at MGH. Females will
be excluded for homogeneity of the sample and because males have higher incidence of autism
than females (4:1). We will seek to enroll up to 50% of the subjects having a history of
positive behavioral effects of fever, which will be recorded from caregivers' recall of
incidents and graded on the CGI-Improvement (CGI-I) 7-point scale.

There will be in total 7 study visits for each subject: the screening visit, enrollment
visit, a blood draw visit at 24 hours after the first dose of study medication (for
mitochondrial/heat shock protein analysis), 4 week (follow-up) visit, 10 week (follow-up)
visit, 18 week visit (last treatment visit), and the final closeout visit one month after the
study drug stops (22 weeks). Even though the treatment will stop at 18 weeks, we will follow
subjects for additional 4 weeks after study medication stops (the 22 week visit) to ensure
safety after study drug stops. Additional visits may be conducted in case any side effects
are reported at any stage of the study.

Inclusion Criteria:

- Autism diagnosis. Quantitative autism traits and severity for diagnosis of autism will
be assessed using the ADOS-G (Modules 1-4 and Severity), Social Responsiveness Scale
(SRS; child and adult forms), Clinical Global Impression-Severity (CGI-S) and Aberrant
Behavior Checklist-Withdrawal subscale (ABC-W).

Exclusion Criteria:

- Absence of a parent or legal guardian and consent

- Unavailability for all visits and adherence to study regimen

- Seizure within 2 years of screening

- Impaired renal function (serum creatinine > 1.2 mg/dl), impaired hepatic function
(AST/ALT > 2x upper limit of normal), impaired thyroid function (TSH outside normal
limits)

- Current infection or treatment with antibiotics; AND

- Chronic medical disorder (e.g., cardiovascular disease, stroke or diabetes) or major
surgery within 3 months prior to enrollment.

- A diagnosis of autism spectrum disorder other than autism, for example, Asperger's,
PDD-NOS etc.