REVIEW
Autism Spectrum Disorders:
Potential Neuro-Psychopharmacotherapeutic
Plant-Based Drugs
Kendy Eduardo Urdaneta,1,* Marı´a Andrea Castillo,1,*
Nola Montiel,2 Neomar Semprún-Hernández,1
Nicola Antonucci,3 and Dario Siniscalco4,5
1
Research Division, Autism Immunology Unit of Maracaibo,
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
2
Maracaibo, Venezuela.
Cátedra Libre de Autismo, Vicerrectorado Académico,
Universidad del Zulia, Maracaibo, Venezuela.
3
Biomedical Centre for Autism Research and Treatment,
Bari, Italy.
4 known as herbal medicinal products.2 According to
Department of Experimental Medicine,
University of Campania, Naples, Italy.
5
Italian Group for Studying Autism—GISA, Brescia, Italy.
*Present address: Department of Biology, Experimental Faculty of
Sciences, University of Zulia, Maracaibo, Venezuela.
ABSTRACT
Over the years, scientific researches have validated the healing
benefits of many psychopharmacotherapeutic plant-based drugs
to ameliorate psychiatric disorders. In contrast, the use of chemical
procedures to isolate and purify specific compounds from plants
that have been used to treat autism spectrum disorders (ASDs) and
its clinical features may contribute to improve the quality of life
of many patients. Also, herbal pharmacological treatments could
improve the core symptoms of autism with fewer side effects. This
review will focus on the uses and actions of phytopharmaceuticals
in the behavioral conditions of ASDs. A large number of natural
compound-based plant drugs have been tested in murine models
of autism and in clinical trials with remarkable success in reversing
the core and associated behaviors with autism such as flavonoids,
cannabinoids, curcuminoids, piperine, resveratrol, and bacosides.
This plant-based drug alternative is safer given that many psy-
chiatric disorders and neurodegenerative pathologies do not often
respond well to currently prescribed medications or have signifi-
cant side effects. However, it is noteworthy to consider the need
for large clinical trials to determine safety and efficacy. Many
results are based on case reports or small size samples, and often
the studies are open label. Standardization of procedures (i.e.,
purity and concentrations) and quality controls are strictly re-
quired to ensure the absence of side effects.
DOI: 10.1089/adt.2018.848 ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018 ASSAY and Drug Development Technologies
Keywords: autism, flavonoids, cannabinoids, bacosides,
piperine
PHYTOPHARMACEUTICALS IN THE MODULATION
OF HUMAN BEHAVIOR
T
he seeking of natural compound plant based drugs
(phytopharmaceuticals) increased significantly in
recent years.1 Phytopharmaceutical drugs are also
U.S. Food and Drug Administration Botanical Drug Develop-
ment Guidance, a botanical product could be classified as a
food (including a dietary supplement), drug (including a bi-
ological drug), medical device, or cosmetic under the Federal
Food, Drug, and Cosmetic Act.3 According to European Union
legislation, the term ‘‘herbal medicinal products’’ are any
medicine that contains a herbal base, one or more herbal
preparations, or one or more of these herbal substances in
combination with one or more such herbal preparations
(DIRECTIVE 2004/24/EC).
Among the different diseases in which research on phyto-
pharmaceuticals has been focused, special emphasis is given
to the herbal extracts able to modulate the human behaviors.
For example, Salvia sp. oil extracts have proven to be efficacy
on cognitive ability, memory, and cognition.4–6 Salvinorin A
is the major component of Salvia divinorum,7 and it has an-
tinociceptive and antidepressant activities8; however, it also
has intense hallucinogenic and dissociative effects.9 It has
demonstrated the efficacy of three herbal extracts (lavender
oil, Hypericum extract, and Ginkgo biloba extract) in treating
anxiety, depression, and dementia, in elderly patients.10
Among the plant-derived phytochemicals, Sage (Salvia la-
vandulaefolia/officinalis), Lemon balm (Melissa officinalis),
and Rosemary (Rosmarinus officinalis) show the most
cognition-enhancing properties.11 Ginseng extract in more
than 4% ginsenosides per oral dose also improves attention
deficit, information processing, cognition, auditory reaction
time, social skills, and mental health.12 Lemon balm shows
mood modulation ability, as it is also able to ameliorate the
negative induced mood effects.13
433
 URDANETA ET AL.
In dementia patients, essential oils have effects on agitation
and aggression.12 It is very interesting that the essential oil of
M. officinalis (Lemon balm), as aromatherapy, is able to re-
duce the agitation in people affected by severe dementia.14
Hypericum perforatum (or St. John’s wort, from its popular
name) has shown some efficacy in treating depression; in
terms of efficacy and adverse effects, it has the same molec-
ular target of selective serotonin reuptake inhibitors in pa-
tients with chronic depressive disorder.15 Piper methysticum
(kava) has been effectively used in treating generalized anx-
iety disorders.16 Kava also shows efficacy against obsessive-
compulsive, phobic, and panic disorders, whereas Passiflora
incarnata (Passionflower), Scutellaria lateriflora (Skullcap),
and Zizyphus jujuba ( Jujube) have been used for anxiety
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
disorders.17 The green tea extracted polyphenol catechin is
able to protect against reduced cognitive functions related to
aging.18 Likely, resveratrol (RSV) and curcumin are the best
known phytochemicals used in cognitive decline.19
Yokukansan, a traditional herbal medicine in Japan, is ef-
fective in treating behavioral and psychological symptoms
associated to dementia.20 Fufangdanshen (Radix Salviae
miltiorrhizae) tablets, a traditional Chinese medicine used to
improve memory, have been positively investigated in treat-
ing cognitive symptoms in vascular dementia patients.21 In a
research of mild cognitive impairments, 95 patients were
treated with Dangguijagyag-san and showed improvements in
cognitive skills without adverse events.22 Several phyto-
pharmaceuticals have been proposed to treat brain inflam-
mation. This process is common and highly prevalent in
psychiatric disorders, such as depression and autism.23 The
anxiolytic activity of Matricaria recutita, G. biloba, Passiflora
incarnata, and S. lateriflora and antidepressant effects of
Echium amoenum, Crocus sativus, and Rhodiola rosea have
been investigated in several clinical trials, showing prelimi-
nary ameliorating effects, without the risk of harmful side
effects.24,25 A new branch or research area is establishing: the
herbomics,24 the complex, genetic, and epigenetic modes of
action of the herbal compounds.
Valerian (Valeriana officinalis) has been proposed for the
treatment of anxiety.26 Moreover, valerian root in association
with Lemon balm is effective in treating attention deficit
disorders, hyperactivity, and impulsiveness in children.27 This
mixed compound is able to improve concentration, hyperac-
tivity, and impulsiveness in primary school children.28 It has
been proposed that the main constituents of valerian, the
valerenic acid and valerenol, are able to bind a specific site on
gamma-aminobutyric acid receptors with agonist activity.29 It
has also been demonstrated that valerian was most effective in
reducing sleep problems in children with hyperactivity.30
434 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018
Sleep problems, anxiety, and hyperactivity are linked to
autism spectrum disorders (ASDs), a neurodevelopmental dis-
order characterized by stereotyped behaviors, language defi-
cits, and difficulties in communication and social interaction.31
Considering that several neurodegenerative diseases and psy-
chiatric disorders are usually poorly responsive to the current
prescribed drugs or have important side effects,32 the herbal
therapies/supplementations could enhance the therapeutic ef-
fect of drug treatments with less adverse events.33 Therefore,
this review will focus on uses of the phytopharmaceuticals and
actions in behavioral conditions of ASDs.
PHYTOPHARMACEUTICALS AND ASDs
Flavonoids
Some individuals with ASDs have been exposed to envi-
ronmental pro-oxidant factors such as heavy metals, pharma-
ceutical compounds (thalidomide, valproic acid, and retinoic
acid), air pollutants, chemical and toxins, and bacterial and viral
infections; those might be the triggering of oxidative stress in
autism.34 To decrease levels of oxidative stress, green tea ther-
apy using Camellia sinensis extract has been proposed. This
plant is an important dietary source of polyphenols, specifically
flavonoids. The main flavonoids present in C. sinensis extract
include catechins (flavan-3-ols) such as epigallocatechin-3-
gallate, epigallocatechin, epicatechin-3-gallate, and epica-
techin; it also contains gallic acid, chlorogenic acid, caffeic acid,
and flavonol derivatives such as kaempferol, myricetin, and
quercetin, which are other constituents of green tea.35,36
The chemical structure of the polyphenols allows them to
cross through the blood–brain barrier,37–47 and its intake has
been associated with neuronal protection against toxicant
agents’ impact. Green tea research shows an amelioration of
behavioral aberrations by a daily intake dose of the extract;
doses range from 75 to 300 mg/kg after postnatal day 14 up to
day 40 in animals with valproate induced autism.37 Green tea
effects have been well documented; due to upregulation
mechanism of inhibitory neurotransmitters, it shows anxio-
lytic and sedative effects; this might be possible by neuro-
modulation of dopamine and serotonin in specific brain areas,
increase in N-Methyl-D-aspartate (NMDA)-independent CA1-
LTP (cornu Ammo 1 Pyramidal Neuro- long-term potentiation),
and decrease in NMDA-dependent CA1-LPT. Besides, green tea
extract decreases tumor necrosis factor-alpha (TNF-a) and C-
reactive protein, both active cytokines in the acute-phase in-
flammatory process; this strongly suggests an anti-inflammatory
mechanism of action.48
A green tea study using mice pups performed behavioral tests
to assess nociception activity, motor coordination, exploratory
ª MARY ANN LIEBERT, INC.

activity, anxiety disorders, and cognitive skills, in which human
doses are based in body surface area calculation,49 with doses
from 6.08 mg to 24.32 mg/kg. Green tea doses might be used
as dietary supplement due to its clinical recommendation as
preventive. This study shows significant results in the im-
provement of behavioral assessments especially with
300 mg/kg of green tea extract; histological findings reveal
the presence of a distinct Purkinje layer and cells after
treatment with green tea, and this confirms that green tea at
that dose has neuroprotective effects.
The cytoprotective action that green tea exerts in neuronal
cells can be asserted and could be efficacious in the treatment of
autism through early intervention with dietary substances.50
Among the adverse effects of green tea, it has been found that
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
higher doses should not be taken with other drugs metabolized
by CYP1A2, CYP2C9, CYP3A4, CYP1A1, CYP2B6, CYP2C8, and
CYP2D6.51,52 The mechanism that explains the hepatotoxicity
of catechins of green tea remains unclear, but factors related to
the patient are becoming predominant.53
In recent years, ASD cases have been increasing from 1/20
live births in 200654 up to 1/68 live births in 201455; none-
tiological causes have been determined yet, which hampered
the development of treatments that contributes to alleviate
ASD symptoms. Nowadays, phytopharmaceutical treatments
have been tried out in clinical trials; one of the flavones that
shows anti-inflammatory, antioxidant, antiallergy, and neu-
roprotective effects is Luteolin (NeuroProtek) with a mixture
of the flavonoids Quercetin and Rutin, which are structurally
related in a liposomal formulation,56 these compounds have
potent inhibition of histamine, interleukin (IL)-6 from mi-
croglia,44 IL-8, TNF, and tryptase release from human mast
cells,57 and they also block mercury-induced cytokine release
from human mast cells.58,59
Luteolin formulation (NeuroProtek) research in children
with ASD states the improvement of ASD symptoms in those
who used the formula for at least 4 months; 75% (28/37) of
patients reported significant improvement in features such as
their stool shape, smell, form, color, and habits within a period
of 2 to 3 weeks. In most of the children (19/37) ‘‘Allergic-like’’
skin symptoms were also significantly reduced. Patients also
improved in about 50% eye contact (15/37) and attention (12/
37). Besides, 30%–50% of patients showed learned tasks (15/
37) and social interactions (14/37) as significant improvement
results. Remarkably, children started speaking words or short
sentences in 10% of patients (4/37). No improvements in
hyperactivity or aggressiveness were registered. Indeed, hy-
peractivity increased in 2/37 children and decreased after 2
weeks with lower doses.56 Based on this evidence researchers
hypothesize if autism is kind of a brain allergy?60
ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018
PHYTOPHARMACEUTICALS AND ASD
Anti-inflammatory flavonoid open-label pilot study as-
sessed a formulation that contains Luteolin, Quercetin, and
Rutin, which is a form of quercetin glycoside in concentra-
tions of 100/70/30 mg/capsule, respectively, extracted from
Chamomile and Sophora japonica leaf. This formula shows its
behavioral effects in children with autism (n = 50) enrolled for
26 weeks, assessing the effectiveness and tolerability. Sig-
nificant improvements were registered in adaptive function-
ing such as communication, daily living, and social skills
( p < 0.005). In addition to the reduction (26.6%–34.8%) in
Aberrant Behavior Checklist subscale scores in overall be-
havior, there was also a transitory increased irritability from 1
to 8 weeks in 27/50 participants.61
Another anti-inflammatory lipid molecule with activity in
acute is N-Palmitoylethanolamide (PEA), which behaves as a
signaling molecule.62 There are a few studies dealing with
phytopharmaceuticals in ASDs; another experimental re-
search asserts the association of this ultramicronized lipid
with Luteolin (co-ultraPEA-LUT) in a murine model of
autism, which is efficient in ameliorating nonsocial autistic
symptoms. It also reduces pro-inflammatory molecules such
as GFAP, Nitrotyrosine, and nuclear factor kappa B [NF-kB],
increases neuroplasticity and neurogenesis, and modulates the
apoptosis mechanism in several brain regions (cerebellum and
hippocampus).63 The results of the assess co-ultraPEA-LUT
assert this lipid combination as a potential biomedical therapy
in the treatment of autism.
It has been reported that luteolin, to exert a hepatotoxic role,
needs a connection with the cytochrome CYP3A-mediated re-
active metabolite formation, and the direct glutathione deple-
tion is an initiating event in luteolin mediated cytotoxicity in
hepatocytes in murine models.64 In addition, previous studies
have reported that luteolin can trigger the mitochondrial
pathway of apoptosis by increasing the activity of caspases.65
Cannabinoids
Recently, the endocannabinoid (EC) system has gained
great interest in ASD research.66 This endogenous system of
signaling molecules consists of arachidonic acid compounds
derived, associated enzymes, and receptors.67 The main
compounds are 2-AG and AEA, which are able to bind and
activate the G-protein-coupled cannabinoid receptors type-1
and type-2 (CB1 and CB2). Once activated, these receptors
block the adenylate cyclase enzyme68 and the following de-
crease of the second messenger molecule cAMP concentra-
tions inside the cell. The EC system shows a key role in ASDs.69
It has been demonstrated that CB2 is overexpressed on
autistic-derived peripheral blood mononuclear cells.70 This
upregulation occurs both at RNA and protein levels, and the
ASSAY and Drug Development Technologies 435
 URDANETA ET AL.
fact that monocytes orchestrate immune responses could in-
dicate that the EC system leads to immunological changes in
ASDs.69 In addition, EC system also plays an important role in
regulating other metabolic pathways involved in ASDs, such
as energy metabolism, neuroinflammation, and food in-
take.71,72 Indirect CB2 stimulation with the use of palmitoy-
lethanolamide has been proven to reduce autistic behaviors in
an animal model and in two case reports.63,73
These data also open the way for a phytopharmaceutical
control of the EC system in ASDs. However, as this system
shows a role in the central nervous system (CNS) development,
its activation might trigger long-lasting functional alter-
ations.74 The famous phytocannabinoid tetrahydrocannabi-
nol (THC), the principal psychoactive extract constituent from
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
Cannabis sativa and Cannabis indica, could trigger persistent
alterations in the still-maturing brain.75 Moreover, the non-
psychoactive phytocannabinoid Cannabidiol (CBD) could
promise therapeutic options as immunomodulation, antioxidant
defense, neuroprotection, and with no side effects.69 CBD has
low affinity for CB1 and CB2, but at different concentrations it
is able to activate a plethora of cellular processes.76 Some au-
tism specialized physicians report CBD oil use to decrease ASD
symptoms.
The use of cannabis-derived compounds for ASD treatment
has been hypothesized some years ago77 and gave much at-
tention also in general media.78 Indeed, the study to assess
efficacy and safety of cannabinoids with a 20:1 ratio of CBD
and THC in amelioration of behavioral alterations in children
and youth with ASD has been approved; other trials aim to
study the safety and efficacy of cannabidivarin (CBDV)
(weight-based dosing of 10 mg/(kg$d) of CBDV for 12 weeks)
in children with autism. CBDV, homolog to CBD, is another
nonpsychoactive compound found in Cannabis; it differs from
this by the side-chain shortened by two methylene bridges
(Clinicaltrials.gov). Before recommending the use of EC drugs
for the amelioration of core ASD symptoms, clinical trials
need to be performed to establish safety, tolerability, and
beneficial effects.69
Piperine
The World Health Organization reported that medicinal
plants are the best source of drugs (WHO, 2010).79 The major
alkaloid commonly used for seizure disorders present in Piper
longum and black pepper Piper nigrum is piperine. It has been
shown to own antioxidant,80 neuroprotective,81 anxiolytic, and
cognition enhancing effects.82 A study reports amelioration
effects of piperine on behavioral alterations and oxidative
stress markers in autism induced murine model, revealing that
piperine treatment restored the motor deficits and decreased the
436 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018
reorientation time, due to its capability to struggle with the
induced cerebellar damage by sodium valproate.
Piperine also has neuroprotective effects on glutamate at
concentration of 20 mg/kg, which induced cell viability res-
toration.83 Oxidative stress alterations might be meaningfully
reversed by treatment with piperine83; besides, a neurobio-
logical finding in ASD is the restoration of the integrity of the
cerebellum by a decrease in number of Purkinje cells, which
are connected with cerebral cortex and limbic system.
Therefore, autism could be a pharmacological condition for
biomedical treatment with piperine.82
Resveratrol
Recently, studies assert that RSV has a neuroprotective role
such as anti-inflammatory and antioxidant effects in many
diseases. RSV (3,5,4-trihydroxy-trans-stilbene) is a naturally
occurring polyphenolic compound present in grapes, pines,
peanuts, and red wine,84 such as all compounds described before
these biological activities could be interesting in the treatment
of ASDs.85 Some factors such as xenobiotics and valproic acid
might trigger or play an important role in the development of
ASD core features.86 Hence, molecular pathway research about
RSV and autism treatment could result as relevant for amelio-
rating core symptoms of autism.87 In humans, the RSV metab-
olites half-life in serum is 9.72 h88 at concentrations of 20 and
40 mg/kg89 or 5, 10, 15 mg/kg per oral.90
A study reveals in experimental murine model that RSV
improves social skills using a rate of 10:270 of RSV:VPA, the
interaction of these molecules enhanced by its physicochemical
features might have an indirect effect at cellular levels87; RSV
regulates and activates sirtuins that are members of the class-III
histone deacetylases.91 It is also known that RSV exerts neu-
roimmunomodulatory effects by regulating Th1/Th2/Th17/Treg
transcription factor signaling,89 decreasing pro-inflammatory
molecules (IL-6 and TNF-a) on dopaminergic neurons,92 inhi-
bition of the NF-kB activation,93 suppression of T cells such as
Treg,94,95 and inhibition of IL-17A.89 Another study supports that
RSV suppresses neuroinflammation in ASDs, finding that RSV
suppresses mitochondrial dysfunction, oxidative-nitrosative
stress, and TNF-a expression in induced rats with ASDs. Hence,
the efficacy of RSV could be as a potential therapeutic agent for
ameliorating neurobehavioral and biochemical alterations re-
lated with ASDs.89
Curcumin
Indian spice turmeric (Curcuma longa) is well known for its
protective effects against neurodegenerative diseases and
neuropsychiatric disorders,96 in which the major curcuminoid
is curcumin (diferuloyl methane), a nontoxic molecule, able to
ª MARY ANN LIEBERT, INC.

cross blood–brain barrier.97 Also, it is reported to have posi-
tive effects on the treatment of autism as curcumin targets
several cell signaling pathways, and its effects are as follows:
increasing intracellular levels of glutathione, reducing in-
flammatory components, mitochondrial dysfunction, oxida-
tive/nitrosative stress, and protein aggregation, counteracting
the damage caused by heavy metals, and supporting liver
detoxification.98–99
Signs of brain toxicity and delayed maturation of brain have
been studied in induced autism animal models; curcumin can
ameliorate these alterations and is able to improve abnormal
brain weight and delayed maturation.100 Pharmacokinetic as-
sessments have showed that curcumin bioavailability is high
and plasma levels of it increased to their highest levels (184 ng/
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
mL) 80 min after oral administration; amorphous solid disper-
sion of curcumin has demonstrated enhanced bioefficiency in
10-fold lower dose.101 Daily doses of 50/100/200 mg/kg of
curcumin regularly given over a period of 4 weeks were dis-
covered to restore neurological, behavioral, biochemical, and
molecular changes related to the ASD phenotype in murine in a
dose-dependent manner. As such, curcumin can be developed as
a neuro-psychopharmacotherapeutical drug due to its charac-
teristics and potential effects for ASD treatment.99 Curcumin
shows ambivalent effects of both genotoxicity and anti-
genotoxicity, endothelial growth, and development in tissue
alterations. Concentration-dependent effects, as low as 0.1 mM
of curcumin, lead to disproportionate DNA segregation, kar-
yorrhexis, and micronucleation in proliferating endothelial
cells.102–104
Bacosides
Bacosides are medicinal substances widely used by Indian
tribes and are the main bioactive compounds extracted from
Bacopa monnieri (L.) Wettst.105 This perennial creeping herb
belongs to the family Scrophulariaceae and is extensively
cultivated in Asia, including China and India. B. monnieri has
numerous synonyms such as brahmi (derived from the word
‘‘Brahma’’), Herpestis monniera, and Gratiola monniera.106 It
has been used for *3,000 years in the Ayurvedic system of
traditional Indian medicine and is classified as a medhyar-
asayana, a drug used to improve cognition and intellect107
and used as a nerve tonic.108
Two triterpenoid glycosides denoted as bacoside A109 and
bacoside B110 are the major bioactive constituents found in B.
monnieri; both of them are able to improve cognition.109,110
Pharmacological effects are attributed to the number of al-
kaloids, saponins, and sterols, which compose the extract.111
It has been reported that these bacosides modulate cholinergic
densities93 along with acetylcholine levels,112 and in presence
ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018
PHYTOPHARMACEUTICALS AND ASD
of these compounds, the central nervous system shows b-
amyloid scavenging properties113 and anxiolytic reliving
processes.114–116 Although its biological molecular mecha-
nisms have not been proven, it is well known that bacoside-A
interferes with aggregation and membrane activity in frag-
ments of the prion protein 38; induction of fibril formation
corresponding to inhibition of membrane interactions is likely
the main factor that reduces amyloid protein toxicity by
bacoside-A.117,118
Even though there are trials based on attention-deficit
hyperactivity disorder (ADHD),119 which is a comorbidity of
ASDs, little research has focused on the efficacy of bacosides
in this population. There is only one report made up to date in
murine models with induced ASDs120 where they evaluate
the effect of B. monnieri extract (300 mg/kg/p.o.) by pow-
dering the whole plant for rat consumption. Behavioral tests
such as nociception, locomotor activity, exploratory activ-
ity, anxiety, and social behavior were performed. Treatment
with B. monnieri significantly improved behavioral alter-
ations and decreased oxidative stress markers; it signifi-
cantly reduced pain threshold and normalized the locomotor
activity and anxiety. Authors inferred that protective ef-
fect of B. monnieri on locomotor activity may be due to its
antianxiety activity and that it decreases accumulated glu-
tamate. Their rats with ASDs also showed neuronal degen-
eration and chromatolysis, and after treatment they observed
its neuroprotective effect with histopathological changes in
cerebellum. A single oral administration of B. monnieri ex-
tract at doses of 30, 60, 300, 1,500, and 5,000 mg/kg given
for large time (270 days) did not produce any toxicity in
murine models.121
Other Compounds with Phytopharmaceutical Relevance
Different types of phytopharmaceuticals and extracts have
been studied briefly on the fields of ASD and ADHD (Table 1).
A study with three ASD patients evaluated a standardized ex-
tract from the ground-up leaves of G. biloba denominated EGb
761, which was administered daily for 4 weeks (2 · 100 mg).
Weekly ratings were performed using Aberrant Behavior and
Symptom Checklist (ABC)122 with clinician ratings consisting
of the Global Assessment Scale,123 Comprehensive Psychiatric
Rating Scale (CPRS),124 and Clinical Global Improvement,125
which were assessed at the beginning and then weekly up to the
end of the treatment period. This treatment improved slightly
ABC factors such as irritability, hyperactivity, inadequate eye
contact, and inappropriate speech. Although they showed a
moderately therapeutic effect of G. biloba in treatment of core
autism symptoms, their results showed no significant im-
provement on the clinician ratings.
ASSAY and Drug Development Technologies 437
 Table 1. Annex; A Summary of Psychopharmacotherapeutic Plant-Based Drug Studies Used in the Treatment
of Autism Spectrum Disorders
Group Phytopharmaceutical
Flavonoids GTE (Camellia sinensis)

Luteolin (NeuroProtek ) from Chamomile sp. and
quercetin Sophora sp. in a formulation of olive
kernel oil
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
Luteolin from Chamomile sp. and quercetin
glycoside rutin from Sophora japonica leaf in a
formulation of olive kernel oil
Palmitoylethanolamide/Luteolin (co-
ultramicronized PEA-LUT)
Standardized extract from leaves of Ginkgo biloba
(EGb 761)
Standardized G. biloba extract (Ginko T.D.)
Alkaloids Synthesized Piperine
438 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018
Participants Results Reference
Murine model, 5 groups with 12 mice each, n = 60. 300 mg/kg of GTE showed improvement in be- Banji et al.36
Group 1 = negative control. havioral assessments, neuroprotective effects, and
Group 2 = treated with GTE alone each day. lowered oxidative stress markers.
Group 3 = treated with VPA.
Group 4 = treated with VPA and received 75 mg/kg
of GTE daily.
Group 5 = treated with VPA receiving 300 mg/kg of
GTE daily.
ASD children, n = 37 treated with 1 capsule/10 kg Improvements in gastrointestinal and allergy Theoharides
made of luteolin (100 mg/capsule), quercetin symptoms (75%), eye contact and attention (50%), et al.56
(70 mg/capsule), and the quercetin glycoside rutin social interaction (25%), speech (10%).
(30 mg/capsule).
ASD children, n = 50 treated with 1 capsule/10 kg Improvement in adaptive functioning and in the Taliou et al.61
made of luteolin (100 mg/capsule), quercetin overall behavior.
(70 mg/capsule), and the quercetin glycoside rutin
(30 mg/capsule).
Murine model, 2 sets of 4 groups, for each group Reduced the expression of pro-inflammatory Bertolino
n = 30 markers, modulated apoptosis in hippocampus and et al.63
I: Group 1 = Sham + Vehicle: cerebellum, increased neurogenesis and neuro-
Group 2 = Sham + co-ultraPEA-LUT (1 mg/kg). plasticity in the hippocampus.
Group 3 = VPA + Vehicle. Improved the clinical status in an ASD child and a
Group 4 = VPA + co-ultraPEA-LUT (1 mg/kg). reduction in motor stereotypes.
II: Group 1 = Sham + Vehicle;
Group 2 = Sham + co-ultraPEA-LUT (1 mg/kg).
Group 3 = VPA + Vehicle.
Group 4 = VPA + co-ultraPEA-LUT (1 mg/kg).
ASD child, n = 1 treated with co-ultraPEA-LUT
(700 mg +70 mg for 1 year).
ASD patients, n = 3 Slight improvement in ABC factors such as Niederhofer146
Treated with EGb 761 (2 dosage · 100 mg daily). irritability, hyperactivity, inadequate eye contact,
and inappropriate speech. Modest therapeutic
effect.
No significant improvement on the clinician
ratings.
ASD outpatients, 2 groups, n = 47 Adding G. biloba to risperidone did not affect the Hasanzadeh
Group 1 = treated with risperidone + Ginko T.D ASD treatment outcome. et al.131
(80 mg/day under 30 kg and 120 mg/day above
30 kg).
Group 2 = risperidone (1–3 mg/day) + placebo.
Murine model, 5 groups with 6 mice each, n = 30. Improved behavioral alterations, lowered oxidative Pragnya
Group 1 = negative control. stress markers, and restored histoarchitecture of et al.83
Group 2 = treated with piperine (20 mg/kg) each cerebellum.
day.
Group 3 = treated with VPA (400 mg/kg).
Group 4 and 5 = a disease treated group received
VPA (400 mg/kg) + piperine (5 and 20 mg/kg).
(continued)
ª MARY ANN LIEBERT, INC.
 Table 1. Continued
Group Phytopharmaceutical Participants Results Reference

Polyphenols RSV dissolved in DMSO (Sigma-Aldrich) Murine model, 4 groups, n = 43. Prenatal administration of RSV prevented the VPA- Bambini-
Group 1 = negative control, n = 7. induced social impairments. Juniors et al.87
Group 2 = treated with RSV each day (3.6 mg/kg),
n = 7.
Group 3 = treated with VPA, n = 19.
Groups 4 and 5 = treated with VPA and RSV,
n = 10.

RSV (Sigma-Aldrich) Murine model, 2 groups. Substantial induction of Foxp3+ and reduction of Bakheet
Group 1 = negative control, received 1% DMSO in T-bet+, GATA-3+, and IL-17A+ expression in CD4+. et al.89
saline only. Upregulated expression of Foxp3 mRNA and
Group 2 = B6 + BTBR group treated with RSV (20 decreased expression levels of T-bet, GATA-3,
or 40 mg/kg). RORgt, and IL-17A in the spleen and brain tissues.
Decreased the protein expression of T-bet, GATA-3,
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.

RORg, and IL-17 and increased Foxp3 in B6 and

BTBR mice.

RSV (Sigma-Aldrich) Murine model, 5 groups with 5 mice each, n = 25.
Group 1 = negative control.
Group 2 = treated with PPA.
Groups 3–5 = ASD mice treated with 5 mg/(kg$d),
10 mg/(kg$d), and 15 mg/(kg$d) of RSV.
Restored the core and associated symptoms of Bhandari and
autistic phenotype by suppressing oxidative- Kuhad90
nitrosative stress, mitochondrial dysfunction, and
TNF-a and MMP-9 expression.

Curcumin (Sigma-Aldrich) Murine model, 8 groups with 5 mice each, n = 40. Restored the core and associated symptoms of Bhandari and
Group 1 = negative control. autistic phenotype by suppressing oxidative- Kuhad99
Group 2 = sodium acetate control (4.0 mL of 1M nitrosative stress, mitochondrial dysfunction, and
solution in PBS). TNF-a and MMP-9 expression.
Group 3 = PBS control (4.0 mL PBS).
Group 4 = propanol control (4.0 mL of 1M Propanol
in PBS).
Group5 = induction group treated with 1M PPA
(4.0 mL in PBS).
Group 6 = ASD induced animals treated with
curcumin 50 mg/(kg$d).
Group 7 = ASD induced animals (1M PPA, 4.0 mL in
PBS) treated with curcumin 100 mg/(kg$d).
Group 8 = ASD induced animals treated with
curcumin 200 mg/(kg$d).

Bacosides Bacopa monnieri (L.) Wettst extract (powdering Murine model, 3 groups, n = 18. Improved behavioral alterations, decreased oxida- Sandhya
the whole plant) Group 1 = negative control. tive stress markers, and restored histoarchitecture et al.120
Group 2 = VPA mice treated with saline. of cerebellum.
Group 3 = VPA mice treated with extract of B.
monnieri (300 mg/kg)

Ginsenosides KRG Murine model, 2 groups, n = 10. Long-term KRG treatment normalized all the Gonzales
Group 1 = treated with KRG solution (100 mg/kg). behaviors except for motor coordination ability. et al.141
Group 2 = treated with KRG solution (200 mg/kg).

Unknown St. John’s Wort (Hypericum perforatum) ASD patients, n = 3. Modest short-term efficacy in treating irritability. Niederhofer147
Treated with 20 mg/day Stereotypy and inappropriate speech factors im-
proved slightly

ASD, autism spectrum disorder; DMSO, dimethyl sulfoxide; GTE, green tea extract; KRG, Korean Red Ginseng; PBS, phosphate buffered saline; PPA, propanoic acid; RSV,
resveratrol; VPA, valproic acid.

ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018 ASSAY and Drug Development Technologies 439
 URDANETA ET AL.
Another work studied their properties as an add-on therapy
with risperidone to treat ASD symptoms.126 The measurement
tool was again the ABC. The dose of risperidone given by the
researchers for the children between 10 and 30 kg was 2 and
3 mg/day for children weighing above 30 kg, with a placebo
control group also evaluated. Their results were not so dif-
ferent to the aforementioned, having no significant impro-
vement in ABC scores.
G. biloba has been trialed in ADHD population with better
results.127 In 2010, Salehi et al.127 carried out a double-blind
randomized controlled trial with 50 outpatients, which re-
ceived a treatment using tablet of Ginko T.D. at a dose of 80–
120 mg/day depending on weight or methylphenidate at a
dose of 20–30 mg/day depending on weight for 6 weeks. They
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
had significant differences between groups, either for In-
attentive or for Hyperactive/Impulsive subscales. Significant
difference was observed on the reduction of scores of the
Parent ADHD Rating Scale at week 6 compared between
groups, but results suggest that administration of G. biloba has
no comparable efficacy in comparison with methylphenidate
in the treatment of ADHD; however, G. biloba treatment
showed less adverse effects.
This extract has been reported to affect the neurotrans-
mitter system, to enhance cerebral blood flow through ar-
teries, veins, and capillaries,128 to be used mainly for treating
dementias,129 and to have antistress, neuroprotective, anti-
inflammatory,130 and antioxidant properties131 that could
impact ASD features. It also has been demonstrated to im-
prove learning memory and synaptic plasticity supported by
some trials in treatment of cognitive impairment.132,133 One of
the major constituents of the EGb is a terpene trilactone de-
noted as ginkgolide B, which has been proven to possess
neuroprotective effects against cerebral ischemia and other
CNS damage.134 However, the cellular and molecular mech-
anisms of the neuroprotective effects of EGb remain unclear.
Some authors compare its action to piracetam,131 which has
been reported to be an effective therapy for ASDs, given as
monotherapy135 or as an add-on therapy.
Another study with effective results on ADHD treatment
was performed using Panax ginseng extract and the ADHD
Rating Scale.136 Study populations were given an oral ad-
ministration of tablets containing 250 mg P. ginseng extract,
27%–30% ginsenosides, pure for 4 weeks. During treatment,
an improvement was observed for the attention score (drop
from 22 to 18), the hyperactive/impulsive score (drop from 24
to 20), and the total score (drop from 46 to 38).
Keeping up with popular herbal extracts used in both tra-
ditional and research-based medicines, there has been reports
that Korean Red Ginseng (KRG) has antioxidant properties137
440 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018
improves cerebral blood flow,138 and has benefits in learning
and memory,139 and has also been related to the central ner-
vous system.140 In a recent study made in murine models,
authors analyzed the therapeutic potential of KRG in allevi-
ating the neurobehavioral deficits found in mice models with
induced ASDs. The methodology consisted of a daily oral
administration of KRG to the study population until all ex-
periments were complete. Mice behaviors were measured,
including social interaction, repetitive behaviors, locomotor
activity, and other behaviors related to ASDs, which were
normalized after a long-term KRG treatment.141
According to a 2009 study, other herbals such as St. John’s
Wort have been studied to alleviate ASD symptoms or modify
some behaviors. It was an open trial with three ASD individ-
uals being administered 20 mg of St. John’s Wort daily for 4
weeks. Outcome was measured by ABC, Psychiatric Rating
Scale Autism, Anger and Speech Deviance factors, Global
Assessment Scale, and Clinical Global Impressions efficacy.
St. John’s Wort was only modestly effective in the short-term
treatment of irritability in some patients with autistic disorder;
however, stereotypy and inappropriate behavior improved
slightly during treatment.142
FUTURE PERSPECTIVES IN THE DESIGN
OF PHYTOPHARMACEUTICALS IN ASDS
The three main ways of obtaining drugs are currently na-
ture, chemical synthesis, and biotechnology, which allows the
discovery of new compounds by genetic engineering tech-
niques. Currently, a high percentage of medicines used in the
allopathic medicine are extracted from the vegetable king-
dom, and there is a remarkable tendency to raise this figure,
due to the discovery of new active principles of botanical
origin. The synthesis and accumulation of the majority of the
compounds with the medical science present in the plants vary
according to the ecological and environmental factors. To
increase the quality and efficacy of herbal medicines and
adjustments, it is important to establish quality control pro-
tocols and standardization of their active ingredients that can
produce many more drugs aimed at reestablishing minimal or
no side effects. The design process helped by bioinformatics
might help to understand combinations and specific param-
eters much more directly linked to ameliorate core autistic
symptoms through its physiological, biochemical, and genetic
base. In this way, when treating the ‘‘symptoms’’ we will treat
the ‘‘causes’’ with potential recovery of the disorder.
Before claiming enthusiastic results, it is noteworthy to
consider the need for large clinical trials to determine safety
and efficacy. Many results are based on case reports or small
size samples, and often the studies are open label. Two recent
ª MARY ANN LIEBERT, INC.

review articles on effectiveness of herbal remedies showed that
data availability on controlled clinical studies is very lim-
ited and the use of different methodologies increases concerns
about effectiveness of herbal medicines.143,144 However, the
efficacy of herbal-based drugs for the treatment of ASD symp-
toms appears to be encouraging. There is also need for more trials
using ASD animal models. This is an important research variable
given that trials are often performed with small size sample;
heterogeneity of ASD symptoms and the complex etiology of
the condition are potential limitations and obstacles to develop
new drugs.145
This review considers that of all plant-based drugs for neu-
romodulation of core ASD features. The flavonoid luteolin
exhibits promising effects, as well as RSV due to its low toxicity
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
reported, interaction with other drugs, and results with statis-
tical significance. However, luteolin is the only molecule that
has been directly tested in children with ASD in whom the core
ASD behaviors have had clinical improvement. In contrast to
RSV that has not been tested in humans but murine model,
studies reported amelioration of autistic clinical features, in
addition to its immunomodulatory effect. Regarding piperine,
improvement of behavioral alterations, lowered oxidative
stress markers, and restored histoarchitecture of cerebellum
were reported. No toxicity studies have been carried out to date,
limiting the authors to their clinical usage due to the high doses
needed to achieve these effects.
DISCLOSURE STATEMENT
The authors declare no conflict of interest in this article.
REFERENCES
1. Chaves SK, Feitosa CM, da S Araújo L: Alkaloids pharmacological activities—
arpospects for the development of phytopharmaceuticals for neurodegenerative
diseases. Curr Pharm Biotechnol 2016;17:629–635.
2. Bhatt A: Phytopharmaceuticals: a new drug class regulated in India. Perspect
Clin Res 2016;7:59–61.
3. Food and Drug Administration: Botanical drug development guidance for industry.
Available at: http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatory
Information/Guidances/UCM458484.pdf (Last accessed on March 1, 2018).
4. Tildesley NT, Kennedy DO, Perry EK, et al.: Salvia lavandulaefolia (Spanish
sage) enhances memory in healthy young volunteers. Pharmacol Biochem
Behav 2003;75:669–674.
5. Tildesley NTJ, Kennedy DO, Perry EK, et al.: Positive modulation of mood and
cognitive performance following administration of acute doses of Salvia
lavandulaefolia essential oil to healthy young volunteers. Physiol Behav 2005;
83:699–709.
6. Scholey AB, Tildesley NT, Ballard CG, et al.: An extract of Salvia (sage) with
anticholinesterase properties improves memory and attention in healthy older
volunteers. Psychopharmacology 2008;198:127–139.
7. Listos J, Merska A, Fidecka S: Pharmacological activity of salvinorin A, the
major component of Salvia divinorum. Pharmacol Rep 2011;63:1305–1309.
8. Cruz A, Domingos S, Gallardo E, et al.: A unique natural selective kappa-opioid
receptor agonist, salvinorin A, and its roles in human therapeutics. Phytochemistry
2017;137:9–14.
ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018
PHYTOPHARMACEUTICALS AND ASD
9. Maqueda AE, Valle M, Addy PH, et al.: Salvinorin-A induces intense dissociative
effects, blocking external sensory perception and modulating interoception and
sense of body ownership in humans. Int J Neuropsychopharmacol 2015;18:pyv065.
10. Kasper S: Phytopharmaceutical treatment of anxiety, depression, and
dementia in the elderly: evidence from randomized, controlled clinical trials.
Wien Med Wochenschr 2015;165:217–228.
11. Kennedy DO, Scholey AB: The psychopharmacology of European herbs with
cognition-enhancing properties. Curr Pharm Des 2006;12:4613–4623.
12. Perry E, Howes MJ: Medicinal plants and dementia therapy: herbal hopes for
brain aging? CNS Neurosci Ther 2011;17:683–698.
13. Kennedy DO, Little W, Scholey AB: Attenuation of laboratory-induced stress in
humans after acute administration of Melissa officinalis (Lemon Balm).
Psychosom Med 2004;66:607–613.
14. Ballard CG, O’Brien JT, Reichelt K, et al.: Aromatherapy as a safe and effective
treatment for the management of agitation in severe dementia: the results of
a double-blind, placebo-controlled trial with Melissa. J Clin Psychiatry 2002;
63:553–558.
15. Rahimi R, Nikfar S, Abdollahi M: Efficacy and tolerability of Hypericum
perforatum in major depressive disorder in comparison with selective serotonin
reuptake inhibitors: a meta-analysis. Prog Neuropsychopharmacol Biol Psychiatry
2009;33:118–127.
16. Sarris J, Kavanagh DJ: Kava and St. John’s Wort: current evidence for use in
mood and anxiety disorders. J Altern Complement Med 2009;15:827–836.
17. Sarris J: Herbal medicines in the treatment of psychiatric disorders: a
systematic review. Phytother Res 2007;21:703–716.
18. Unno K, Hoshino :. Brain senescence and neuroprotective dietary components.
Cent Nerv Syst Agents Med Chem 2007;7:109–114.
19. Cicero AFG, Fogacci F, Banach M: Botanicals and phytochemicals active on
cognitive decline: the clinical evidence. Pharmacol Res 2018;130:204–212.
20. Soraoka H, Oniki K, Matsuda K, et al.: The effect of Yokukansan, a traditional
herbal preparation used for the behavioral and psychological symptoms of
dementia, on the drug-metabolizing enzyme activities in healthy male
volunteers. Biol Pharm Bull 2016;39:1468–1474.
21. Tian J, Shi J, Wei M, et al.: The efficacy and safety of Fufangdanshen tablets
(Radix Salviae miltiorrhizae formula tablets) for mild to moderate vascular
dementia: a study protocol for a randomized controlled trial. Trials 2016;17:281.
22. Kim KH, Go HY, Lee JA, et al.: The effect of Dangguijagyag-san on mild
cognitive impairment. J Altern Complement Med 2016;22:509–514.
23. Bjorklund G, Saad K, Chirumbolo S, et al.: Immune dysfunction and
neuroinflammation in autism spectrum disorder. Acta Neurobiol Exp (Wars)
2016;76:257–268.
24. Sarris J, Panossian A, Schweitzer I, et al.: Herbal medicine for depression,
anxiety and insomnia: a review of psychopharmacology and clinical evidence.
Eur Neuropsychopharmacol 2011;21:841–860.
25. Lakhan SE, Vieira KF: Nutritional and herbal supplements for anxiety and
anxiety-related disorders: systematic review. Nutr J 2010;9:42.
26. Larzelere MM, Campbell JS, Robertson M: Complementary and alternative
medicine usage for behavioral health indications. Prim Care 2010;37:213–236.
27. Ross SM: Valerian root and lemon balm extracts: a phytomedicine compound
improves symptoms of hyperactivity, attention deficits, and impulsivity in
children. Holist Nurs Pract 2015;29:391–395.
28. Gromball J, Beschorner F, Wantzen C: Hyperactivity, concentration difficulties and
impulsiveness improve during seven weeks’ treatment with valerian root and lemon
balm extracts in primary school children. Phytomedicine 2014;21:1098–1103.
29. Benke D, Barberis A, Kopp S, et al.: GABA (A) receptors as in vivo substrate for
the anxiolytic action of valerenic acid, a major constituent of valerian root
extracts. Neuropharmacology 2009;56:174–181.
30. Francis AJ, Dempster RJ: Effect of valerian, Valeriana edulis, on sleep
difficulties in children with intellectual deficits: randomised trial.
Phytomedicine 2002;9:273–279.
31. Hsiao EY, McBride SW, Hsien S, et al.: Microbiota modulate behavioral and
physiological abnormalities associated with neurodevelopmental disorders.
Cell 2013;155:1451–1463.
ASSAY and Drug Development Technologies 441
 URDANETA ET AL.
32. Barreto GE, Avila-Rodriguez M, Foitzick M, et al.: Advances in medicinal plants
with effects on anxiety behavior associated to mental and health conditions.
Curr Med Chem 2017;24:411–423.
33. Baek JH, Nierenberg AA, Kinrys G: Clinical applications of herbal medicines for
anxiety and insomnia; targeting patients with bipolar disorder. Aust N Z J
Psychiatry 2014;48:705–715.
34. Chauhan A, Chauhan V: Oxidative stress in autism. Pathophysiology 2006;13:
171–181.
35. Cabrera C, Artacho R, Gimenez R: Beneficial effects of green tea. J Am Coll
Nutr 2006;25:79–99.
36. Banji D, Banji OJ, Abbagoni S, et al.: Amelioration of behavioral aberrations
and oxidative markers by green tea extract in valproate induced autism in
animal. Brain Res 2011;1410:141–151.
37. Hong JT, Ryu SR, Kim HJ, et al.: Neuroprotective effect of green tea extract in
experimental ischemia—reperfusion brain injury. Brain Res Bull 2000;53:743–749.
38. Wang J, Ferruzzi MG, Ho L, et al.: Brain-targeted proanthocyanidin metabolites
for Alzheimer’s disease treatment. J Neurosci 2012;32:5144–5150.
39. Vaidyanathan JB, Walle T: Transport and metabolism of the tea flavonoid(-)
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
epicatechin by the human intestinal cell line Caco-2. Pharm Res 2001;18:
1420–1425.
40. Hong J, Lambert JD, Lee S-H, Sinko PJ, Yang CS: Involvement of multidrug
resistance-associated proteins in regulating cellular levels of (-)-
epigallocatechin-3-gallate and its methyl metabolites. Biochem Biophys Res
Commun 2003;310:222–227.
41. Wang S, Chen R, Zhong Z, Shi Z, Chen M, Wang Y: Epigallocatechin-3-gallate
potentiates the effect of curcumin in inducing growth inhibition and
apoptosis of resistant breast cancer cells. Am J Chin Med 2014;42:1279–1300.
42. Shukla S, Zaher H, Hartz A, Bauer B, Ware JA, Ambudkar SV: Curcumin inhibits
the activity of ABCG2/BCRP1, a multidrug resistance-linked ABC drug
transporter in mice. Pharm Res 2009;26:480–487.
43. Mitsunaga Y, Takanaga H, Matsuo H, et al.: Effect of bioflavonoids on vincristine
transport across blood-brain barrier. Eur J Pharmacol 2000;395:193–201.
44. Wu C-P, Calcagno AM, Hladky SB, Ambudkar SV, Barrand MA: Modulatory
effects of plant phenols on human multidrug resistance proteins 1, 4 and 5
(ABCC1, 4 and 5). FEBS J 2005;272:4725–4740.
45. Cooray HC, Janvilisri T, Van Veen HW, Hladky SB, Barrand MA: Interaction of
the breast cancer resistance protein with plant polyphenols. Biochem Biophys
Res Commun 2004;317:269–275.
46. Choi J-S, Choi B-C, Kang KW: Effect of resveratrol on the pharmacokinetics of
oral and intravenous nicardipine in rats: possible role of P-glycoprotein
inhibition by resveratrol. Die Pharmazie-An Int J Pharm Sci 2009;64:49–52.
47. Yang Y, Bai L, Li X, et al.: Transport of active flavonoids, based on cytotoxicity
and lipophilicity: an evaluation using the blood-brain barrier cell and Caco-2
cell models. Toxicol In Vitro 2014;28:388–396.
48. Lardner AL: Neurobiological effects of the green tea constituent theanine and
its potential role in the treatment of psychiatric and neurodegenerative
disorders. Nutr Neurosci 2014;17:145–155.
49. Maiti S, Acharyya N, Gosh TK, et al.: Green tea (Camellia sinensis) protects
against arsenic neurotoxicity via antioxidative mechanism and activation of
superoxide dismutase activity. Cent Nerv Syst Agents Med Chem 2017;17:
187–195.
50. Shaw SR, Nihal M, Ahmad N: Dose translation from animal to human studies
revisited. FASEB J 2008;22:659–661.
51. Satoh T, Fujisawa H, Nakamura A, Takahashi N, Watanabe K: Inhibitory effects
of eight green tea catechins on cytochrome P450 1A2, 2C9, 2D6, and 3A4
activities. J Pharm Pharm Sci 2016;19:188–197.
52. Albassam AA, Markowitz JS: An appraisal of drug-drug interactions with green
tea (Camellia sinensis). Planta Med 2017;83:496–508.
53. Mazzanti G, Di Sotto A, Vitalone A: Hepatotoxicity of green tea: an update.
Arch Toxicol 2015;89:1175–1191.
54. Gurney JG, McPheeters ML, Davis MM: Parental report of health conditions
and health care use among children with and without autism: National Survey
of Children’s Health. Arch Pediatr Adolesc Med 2006;160:825–830.
442 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018
55. Baoi J: Prevalence of autism spectrum disorder among children aged 8 years-
autism and developmental disabilities 617 monitoring network, 11 sites,
United States, 2010. MMWR Surveill Summ 2014;63:1–21.
56. Theoharides TC, Asadi S, Panagiotidou S: A case series of a luteolin
formulation (NeuroProtek) in children with autism spectrum disorders. Int J
Immunopathol Pharmacol 2012;25:317–323.
57. Jang S, Kelley KW, Johnson RW: Luteolin reduces IL-6 production in microglia
by inhibiting JNK phosphorylation and activation of AP-1. Proc Natl Acad Sci
USA 2008;105:7534–7539.
58. Kempuraj D, Madhappan B, Christodoulou S, et al.: Flavonols inhibit
proinflammatory mediator release, intracellular calcium ion levels and protein
kinase C theta phosphorylation in human mast cells. Br J Pharmacol 2005;145:
934–944.
59. Asadi S, Zhang B, Weng Z, et al.: Luteolin and thiosalicylate inhibit HgCl(2) and
thimerosal induced VEGF release from human mast cells. Int J Immunopathol
Pharmacol 2010;23:1015–1020.
60. Theoharides T: Is a subtype of autism an allergy of the brain? Clin Ther 2013;
35:584–589.
61. Taliou A, Zintzaras E, Lykouras L, et al.: An open-label pilot study of a formulation
containing the anti-inflammatory flavonoid luteolin and its effects on behavior in
children with autism spectrum disorders. Clin Ther 2013;35:592–602.
62. Skaper SD, Facci L, Fusco M, et al.: Palmitoylethanolamide, a naturally occurring
disease modifying agent in neuropathic pain. Inflammopharmacology 2014;22:
79–94.
63. Bertolino B, Crupi R, Impellizzeri D, et al.: Beneficial effects of co-
ultramicronized palmitoylethanolamide/luteolin in a mouse model of autism
and in a case report of autism. CNS Neurosci Ther 2017;23:87–98.
64. Shi F, Zhao P, Li X, Pan H, Ma S, Ding L: Cytotoxicity of luteolin in primary rat
hepatocytes: the role of CYP3A-mediated ortho-benzoquinone metabolite
formation and glutathione depletion. J Appl Toxicol 2015;35:1372–1380.
65. Michels G, Wätjen W, Niering P, et al.: Pro-apoptotic effects of the flavonoid
luteolin in rat H4IIE cells. Toxicology 2005;206:337–348.
66. Siniscalco D: Endocannabinoid system as novel therapeutic target for autism
treatment. Autism Open Access 2014;4:e122.
67. Pertwee RG: Endocannabinoids and their pharmacological actions. Handb Exp
Pharmacol 2015;231:1–37.
68. Pertwee RG, Howlett AC, Abood ME, et al.: International union of basic and
clinical pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond
CB1 and CB2. Pharmacol Rev 2010;62:588–631.
69. Brigida AL, Schultz S, Cascone M, et al.: Endocannabinod signal dysregulation
in autism spectrum disorders: a correlation link between inflammatory state
and neuro-immune alterations. Int J Mol Sci 2017;18:E1425.
70. Siniscalco D, Sapone A, Giordano C, et al.: Cannabinoid receptor type 2, but
not type 1, is up-regulated in peripheral blood mononuclear cells of children
affected by autistic disorders. J Autism Dev Disord 2013;43:2686–2695.
71. Siniscalco D, Bradstreet JJ, Cirillo A, et al.: The in vitro GcMAF effects on
endocannabinoid system transcriptionomics, receptor formation, and cell
activity of autism-derived macrophages. J Neuroinflammation 2014;11:78.
72. Habib SS, Al-Regaiey K, Bashir S, et al.: Role of endocannabinoids on
neuroinflammation in autism spectrum disorder prevention. J Clin Diagn Res
2017;11:CE01–CE03.
73. Antonucci N, Cirillo A, Siniscalco D: Beneficial effects of palmitoylethanolamide
on expressive language, cognition, and behaviors in autism: a report of two
cases. Case Rep Psychiatry 2015;2015:325061.
74. Campolongo P, Trezza V, Palmery M, et al.: Developmental exposure to
cannabinoids causes subtle and enduring neurofunctional alterations. Int Rev
Neurobiol 2009;85:117–133.
75. Jager G, Ramsey NF: Long-term consequences of adolescent cannabis exposure
on the development of cognition, brain structure and function: an overview of
animal and human research. Curr Drug Abuse Rev 2008;1:114–123.
76. Pertwee RG: The diverse CB1 and CB2 receptor pharmacology of three
plant cannabinoids: D9-tetrahydrocannabinol, cannabidiol and D9-
tetrahydrocannabivarin. Br J Pharmacol 2008;153:199–215.
ª MARY ANN LIEBERT, INC.

77. Grinspoon L: A novel approach to the symptomatic treatment of autism.
O’Shaughnessy’s. 2010. Available at: http://www.beyondthc.com/wp-content/
uploads/2013/08/GrinspoonAutism.pdf (Last accessed on March 1, 2018).
78. Lawrence Norman R: Future’s mirror: the endocannabinoid system in human
pathology. J Unconscious Psychology. Available at: https://www.haaretz.com/
science-and-health/israeli-doctors-to-study-use-of-cannabis-to-treat-autism-
1.5430382 (Last accessed on March 1, 2018).
79. World Health Organization: Medicinal plants commonly used in the newly
independent states (NIS). Geneve 2010. Available at: http://www.who.int/
medicines/areas/traditional/monograph_eng.pdf (Last accessed on January 2018).
80. Choi BM, Kim SM, Park TK, et al.: Piperine protects cisplatin-induced apoptosis
via heme oxygenase-1 induction in auditory cells. J Nutr Biochem 2007;18:
615–622.
81. Fu M, Sun ZH, Zuo HC: Neuroprotective effect of Piperine on pri-marily
cultured hippocampal neurons. Biol Pharm Bull 2010;33:598–603.
82. Wattanathorn J, Chonpathompikunlert P, Muchimapura S, et al.: Piperine, the
potential functional food for mood and cognitivedisorders. Food Chem Toxicol
2008;46:3106–3110.
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.
83. Pragnya B, Kameshwari JS, Veeresh B: Ameliorating effect of piperine on
behavioral abnormalities and oxidative markers in sodium valproate induced
autism in BALB/C mice. Behav Brain Res 2014;15;270:86–94.
84. Vang N, Ahmad CA, Baile JA, et al.: What is new for an old molecule?
Systematic review and recommendations on the use of resveratrol. PLoS One
2011;6:e19881.
85. Rossignol DA, Frye RE: Evidence linking oxidative stress, mitochondrial
dysfunction, and inflammation in the brain of individuals with autism. Front
Physiol 2014;5:150.
86. Herbert MR: Contributions of the environment and environmentally
vulnerable physiology to autism spectrum disorders. Curr Opin Neurol 2010;
23:103–110.
87. Bambini-Juniora V, Zanattaa G, Della Flora-Nunesa G, et al.: Resveratrol
prevents social deficits in animal model of autism induced by valproic acid.
Neurosc Letters 2014;176–181.
88. Walle T, Hsieh F, DeLegge M, et al.: High absorption but very low bioavailability
of oral resveratrol in humans. Drug Metab Dispos 2004;32:1377–1382.
89. Bakheet S, Zeed M, Ahmad Ansari M, et al.: Resveratrol ameliorates
dysregulation of Th1, Th2, Th17, and T regulatory cell-related transcription
factor signaling in a BTBR T + tf/J mouse model of autism. Mol Neurobiol
2017;54:5201–5212.
90. Bhandari R, Kuhad A: Resveratrol suppresses neuroinflammation in the
experimental paradigm of autism spectrum disorders. Neurochem Int 2017;
103:8–23.
91. Lakshminarasimhan M, Rauh D, Schutkowski M, et al.: Sirt1 activation by
resveratrol is substrate sequence-selective. Aging (Albany) 2013:5:151–154.
92. Zhang F, Liu J, Shi JS: Anti-inflammatory activities of resveratrol in the brain:
role of resveratrol in microglial activation. Eur J Pharmacol 2010;636:1–7.
93. Sha H, Ma Q, Jha RK, et al.: Resveratrol ameliorates hepatic injury via the
mitochondrial pathway in rats with severe acute pancreatitis. Eur J Pharmacol
2008;601:136–142.
94. Wang B, Sun J, Li X, et al.: Resveratrol prevents suppression of regulatory T-
cell production, oxidative stress, and inflammation of mice prone or resistant
to high-fat diet-induced obesity. Nutr Res 2013;33:971–981.
95. Zozulya AL, Wiendl H: The role of regulatory T cells in multiple sclerosis. Nat
Clin Pract Neurol 2008;4:384–398.
96. Mythri RB, Bharath MM: Curcumin: a potential neuroprotective agent in
Parkinson’s disease. Curr Pharm Des 2012;18:91–99.
97. Aggarwal BB, Harikumar KB: Potential therapeutic effects of curcumin, the
anti-inflammatory agent, against neurodegenerative, cardiovascular,
pulmonary, metabolic, autoimmune and neoplastic diseases. Int J Biochem
Cell Biol 2009;41:40–59.
98. Blaylock RL, Strunecka A: Immune-glutamatergic dysfunction as a central
mechanism of the autism spectrum disorders. Curr Med Chem 2009;16:
157–170.
ª MARY ANN LIEBERT, INC.  VOL. 16 NO. 8  NOVEMBER/DECEMBER 2018
PHYTOPHARMACEUTICALS AND ASD
99. Bhandari R, Kuhad A: Neuropsychopharmacotherapeutic efficacy of curcumin in
experimental paradigm of autism spectrum disorders. J Life Sci 2015;141:156–169.
100. Al-Askar M, Bath RS, Selim M, et al.: Postnatal treatment using curcumin
supplements to amend the damage in VPA-Induced rodent models of autism.
BMC Complement Altern Med 2017;17:259.
101. Chuah AM, Jacob B, Jie Z, et al.: Enhanced bioavailability and bioefficacy of an
amorphous solid dispersion of curcumin. Food Chem 2014;156:227–233.
102. Cao J, Jiang L, Liu Y, Yang G, Yao XF, Zhong LF: Curcumin-induced genotoxicity
and antigenotoxicity in HepG2 cells. Toxicon 2007;49:1219–1222.
103. Balaji S, Chempakam B: Toxicity prediction of compounds from turmeric
(Curcuma longa L.). Food Chem Toxicol 2010;48:2951–2959.
104. Jackson SJ, Murphy LL, Venema RC, Singletary KW, Young AJ: Curcumin binds
tubulin, induces mitotic catastrophe, and impedes normal endothelial cell
proliferation. Food Chem Toxicol 2013;60:431–438.
105. Malishev R, Shaham-Niv S, Nandi S, et al.: Bacoside-A, an Indian traditional-
medicine substance, inhibits b-amyloid cytotoxicity, fibrillation, and
membrane interactions. ACS Chem Neurosci 2017;8:884–891.
106. Gohil KJ, Patel JJ: A review on Bacopa monniera: current research and future
prospects. Int J Green Pharm 2010;4:1–9.
107. Russo A, Borrelli F: Bacopa monniera, a reputed nootropic plant: an overview.
Phytomedicine 2005;12:305–317.
108. Mukherjee GD, Dey CD: Clinical trial on Brahmi. J Exp Med Sci 1966;10:5–11.
109. Bacopin: Chemical Constituents. Available at: http://bacopin.com/chemical.htm
(Last accessed on March 1, 2018).
110. United Nations Office for South-South Cooperation: Examples of the
development of pharmaceutical products from medicinal plants. In: U.
Memory Enhancer: Bacopa monniera Standardized Fraction, pp. 29–44.
Lucknow, India, 2003.
111. Kean JD, Kaufman J, Lomas J, et al.: A randomized controlled trial investigating
the effects of a special extract of Bacopa monnieri (CDRI 08) on hyperactivity
and inattention in male children and adolescents: BACHI Study Protocol.
Nutrients 2015;7:9931–9945.
112. Uabundit N, Wattanathorn J, Mucimapura S, et al.: Cognitive enhancement
and neuroprotective effects of Bacopa monnieri in Alzheimer’s disease model.
J Ethnopharm 2010;127:26–31.
113. Bhattacharya SK, Kumar A, Ghosal S: Effect of Bacopa monnieri on animal
models of Alzheimer’s disease and perturbed central cholinergic markers of
cognition in rats. Res Commun Pharmacol Toxicol 1999;4:1–12.
114. Holcomb LA, Dhanasekaran M, Hitt AR: Bacopa monnieri extract reduces
amyloid levels in PSAPP mice. J Alzheimer’s Disease 2006;9:243–251.
115. Bhattacharya SK, Bhattacharya A, Kumar A, et al.: Antioxidant activity of
Bacopa monniera in rat frontal cortex, striatum and hippocampus. Phytother
Res 2000;14:174–179.
116. Kapoor R, Srivastava S, Kakkar P: Bacopa monnieri modulates antioxidant
responses in brain and kidney of diabetic rats. Environ Toxicol Pharmacol
2009;27:62–69.
117. Dhanasekaran M, Tharakan B, Holcomb LA: Neuroprotective mechanisms of
ayurvedic antidementia botanical Bacopa monniera. Phytother Res 2007;21:
965–969.
118. Malishev R, Nandi S, Kolusheva S, et al.: Bacoside-A, an anti-amyloid natural
substance, inhibits membrane disruption by the amyloidogenic determinant of
prion protein through accelerating fibril formation. Biochim Biophys Acta
2016;1858:2208–2214.
119. Jauhari N, Singh YD, Kushwaha KP, et al.: Clinical evaluation of Bacopa
monniera extract on behavioural and cognitive functions in children suffering
from attention deficit hyperactivity disorder. Central Drug Research Institute
(CDRI), Varanasi, India, 2001.
120. Sandhya T, Sowjanya J, Veeresh B: Bacopa monniera (L.) Wettst ameliorates
behavioral alterations and oxidative markers in sodium valproate induced
autism in rats. Neurochem Res 2012;37:1121–1131.
121. Sireeratawong S, Jaijoy K, Khonsung P, Lertprasertsuk N, Ingkaninan K: Acute
and chronic toxicities of Bacopa monnieri extract in Sprague-Dawley rats.
BMC Complement Altern Med 2016;16:249.
ASSAY and Drug Development Technologies 443
 URDANETA ET AL.

122. Aman MG, Singh NN, Stewart AW, et al.: Psychometric characteristics of the 145. Lacivita E, Perrone R, Margari L, Leopoldo M: Targets for drug therapy for
Aberrant Behavior Checklist. Am J Ment Def 1985;99:492–502. autism spectrum disorder: challenges and future directions. J Med Chem
123. Shaffer D, Gould MS, Brasic J, et al.: The Children’s Global Assessment Scale 2017;60:9114–9141.
(CGAS): adapted from Global Assessment Scale for Adults. Arch Gen 146. Niederhofer H: First preliminary results of an observation of Ginkgo biloba
Psychiatry 1973;40:1228–1231. treating patients with autistic disorder. Phytother Res 2009;1645–1646.
124. Overall JE, Campbell M: Behavioral assessment of psychopathology in 147. Niederhofer H: St John’s Wort treating patients with autistic disorder.
children: infantile autism. J Clin Psychol 1988;44:708–716. Phytother Res 2009;23:1521–1523.
125. National Institute of Mental Health: Special feature: rating scales and
assessment instruments for use in pediatric psychopharmacology research.
Psychopharmacol Bull 1985;21:714–1124.
126. Akhondzadeh S, Tajdar H, Mohammadi M, et al.: Double-blind controlled trial Address correspondence to:
of piracetam added to risperidone in patients with autistic disorder. Child Neomar Semprún-Hernández, MS
Psychiatry Hum Dev 2008;39:237–245. Research Division
127. Salehi B, Imani R, Mohammadi M, et al.: Ginkgo biloba for attention-deficit/
hyperactivity disorder in children and adolescents: a double blind,
Autism Immunology Unit of Maracaibo
randomized controlled trial. Prog Neuropsychopharmacol Biol Psychiatry Maracaibo 4001
2010;34:76–80. Venezuela
128. EGb 761: Ginkgo biloba extract, Ginkor. Drugs RD 2003;4:188–193.
Downloaded by 82.145.211.200 from www.liebertpub.com at 01/28/23. For personal use only.

129. Lijuan J, Su L, Cui H, et al.: Ginkgo biloba extract for dementia: a systematic E-mail: neomar.semprun@gmail.com
review. Shanghai Arch Psychiatry 2013;25:10–21.
130. Maclennan KM, Darlington CL, Smith PF: The CNS effects of Ginkgo biloba
extracts and ginkgolide B. Prog Neurobiol 2002;67:235–257.
131. Hasanzadeh E, Mohammadi MR, Ghanizadeh A, et al.: A Double-Blind Placebo Abbreviations Used
controlled trial of Ginkgo biloba added to risperidone in patients with autistic
2-AG ¼ 2-arachidonoyl glycerol
disorders. Child Psychiatry Hum Dev 2012;43:674–682.
ABC ¼ Aberrant Behavior and Symptom Checklist
132. DeFeudis FV, Drieu K: Ginkgo biloba extract (EGb 761) and CNS functions:
ADHD ¼ attention-deficit hyperactivity disorder
basic studies and clinical applications. Curr Drug Targets 2000;1:25–58.
AEA ¼ N-arachidonoylethanolamine (anandamide)
133. Akhondzadeh S: Hippocampal synaptic plasticity and cognition. J Clin Pharm
ASD ¼ autism spectrum disorder
Ther 1999;24:241–248.
CA1-LTP ¼ cornu Ammo 1 Pyramidal Neuro- long-term potentiation
134. Xia SH, Fang DC: Pharmacological action and mechanisms of ginkgolide B.
cAMP ¼ Cyclic adenosine monophosphate
Chin Med J (Engl) 2007;120: 922–928.
CB1 ¼ Cannabinoid receptor 1
135. Paczynski M: Piracetam: a novel therapy for autism? J Autism Dev Disord
CB2 ¼ Cannabinoid receptor 2
1997;27:628–630.
CBD ¼ Cannabidiol
136. Niederhofer H: Panax ginseng may improve some symptoms of attention-
CBDV ¼ cannabidivarin
deficit hyperactivity disorder. J Diet Suppl 2009;6:22–27.
CD4+ ¼ cluster of differentiation 4
137. Lee JW, Mo EJ, Choi JE, et al: Effect of Korean Red Ginseng extraction
CNS ¼ Central Nervous System
conditions on antioxidant activity, extraction yield, and ginsenoside Rg1 and
co-ultraPEA-LUT ¼ co-ultramicronized N-Palmitoylethanolamide-Luteolin
phenolic content: optimization using response surface methodology. J
CPRS ¼ Comprehensive Psychopathological Rating Scale
Ginseng Res 2016;40:229–236.
DMSO ¼ dimethyl sulfoxide
138. Keum YS, Park KK, Lee J-M, et al.: Antioxidant and anti-tumor promoting
EC ¼ endocannabinoid
activities of the methanol extract of heat-processed ginseng. Cancer Lett
Foxp3 ¼ forkhead box P3
2000;150:418.
GATA-3 ¼ Erythroid transcription factor 3
139. Kim CS, Park JB, Kim K-J, et al.: Effect of Korea red ginseng on cerebral blood
GTE ¼ green tea extract
flow and superoxide production. Acta Pharmacol Sin 2002;23:11526.
IL ¼ interleukin
140. Jin SH, Park JK, Nam KY, et al.: Korean red ginseng saponins with low ratios of
KRG ¼ Korean Red Ginseng
protopanaxadiol and protopanaxatriol saponin improve scopolamine-induced
LPT ¼ long-term potentiation
learning disability and spatial working memory in mice. J Ethnopharmacol
NF-kB ¼ nuclear factor kappa B
1999;66:1239.
NMDA ¼ N-Methyl-D-aspartate
141. Gonzales EL, Jang JH, Mabunga DF, et al.: Supplementation of Korean Red
PBS ¼ phosphate buffered saline
Ginseng improves behavior deviations in animal models of autism. Food Nutr
PEA ¼ N-Palmitoylethanolamide
Res 2016;60:29245.
PPA ¼ propanoic acid
142. Gasparotto FM, Dos Reis Lı́vero FA, Tolouei Menegati SEL, Junior AG: Herbal
RORgt ¼ RAR-related orphan receptor gamma
medicine as an alternative treatment in autism spectrum disorder: a
RSV ¼ resveratrol
systematic review. Curr Drug Metab 2018;19:454–459.
Th1/Th2/Th17/Treg ¼ Type 1 T helper/Type 2 T helper/Type 17
143. Bang M, Lee SH, Cho SH, et al.: Herbal medicine treatment for children with
T helper/regulatory T cell
autism spectrum disorder: a systematic review. Evid Based Compl Alternat
THC ¼ tetrahydrocannabinol
Med 2017;2017:12.
TNF-a ¼ tumor necrosis factor-alpha
144. Chadman KK: Animal models for autism in 2017 and the consequential
VABS ¼ Vineland Adaptive Behavior Scales
implications to drug discovery. Expert Opin Drug Discov 2017;12:1187–
VPA ¼ valproic acid
1194.

444 ASSAY and Drug Development Technologies NOVEMBER/DECEMBER 2018 ª MARY ANN LIEBERT, INC.