1. Theoharides C. Theoharides,
Novel Treatment Approaches to Autism: How genes and environment contribute to brain inflammation and symptoms
Theoharides C. Theoharides,
MS, PhD, MD, FAAAAI
Professor of Pharmacology, Internal Medicine and Biochemistry; Associate Professor of Psychiatry
Director, Molecular Immunopharmacology and Drug Discovery Laboratory
Tufts University School of Medicine-Tufts Medical Center
Boston, MA, USA
Clinical Pharmacologist, Massachusetts Drug Formulary Commission ( )

2. Most Common Problems in Children with ASD
Brain inflammation (high IL-6 and TNF)
Blood-brain-barrier disruption
Brain fog
Food allergies/intolerance/GI complaints
Inability to socialize
Lack of eye contact and attention
Loss of speech
Mitochondrial dysfunction
Reduced sociability
Skin eruptions

4. Allergic-Like Symptoms
Allergies
Angioneurotic edema
Atopy
Atopic dermatitis
Eczema
Food allergy
Food intolerance
Idiopathic urticaria
Idiopathic mast activation disorder
Mastocytosis
Mast cell activation syndrome
Non-IgE food allergy
Urticaria pigmentosa
Edvard Munch, The Scream (Skrik, 1893)
Copyright Dr. T.C. Theoharides

6. Copyright Dr. T.C. Theoharides
Mast Cells Are Tissue Immune Cells Involved in All the Pathological Processes Suspected in Autism
Play a role in the allergic response
Upon stimulation release many mediators (ex. Histamine)
Picture:
Mast cells can be activated by many environmental, infectious and stress triggers leading to the release of mediators with potent vasodilatory, nociceptive, and inflammatory properties
Copyright Dr. T.C. Theoharides

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11. The Mast Cell Is the “Canary” of the Body
Mast cell activation by environmental triggers
Bacterial and viral antigens, environmental toxins such as polychlorinated biphenyl (PCB) and mercury, preservatives, toxins.

20. Mercury chloride induces significant VEGF release and augments SP-induced VEGF release
4/22/2017
1a: from 500 to 750 & almost 850, similar to SP.
When combined, to more than 1000.

21. Neurotensin Is a Mast Cell Trigger Present Both in the Brain and the Gut
NT is a peptide originally isolated from the brain.
Most NT receptors in the brain are in the Broca area responsible for language, often lost in ASD children.
NT is neurotoxic and can induce seizures.
NT is present also in the GI tract, where it can induce intestinal inflammation.
NT can activate immune cells, and act together with corticotropin-releasing hormone (CRH), secreted under stress, to stimulate mast cells

22. Donelan et al. PNAS USA 103:7759-7764, 2006.

23. Copyright Dr. T.C. Theoharides
4/22/2017
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24. Copyright Dr. T.C. Theoharides
Funded by Safe Minds and the Autism Collaborative
Angelidou A et al. J Neuroinflammation Aug 23;7:48.
Copyright Dr. T.C. Theoharides

31. Serum mitochondrial DNA and anti-mitochondrial antibodies are present in young children with autism
Funded by the Autism Research Collaborative and Theta Biomedical, Inc.
Zhang et al. J Neuroinflammation. 2010, In press.

33. How environmental and stress factors influence gene activation
Role of Epigenetics
How environmental and stress factors influence gene activation

34. Twin pair study indicates that >60% of influence is not genetic.
72, October 20, 2011
Many gene mutations discovered, but only explain about 5% of ASD cases.
Twin pair study indicates that >60% of influence is not genetic.

36. Cell size & proliferation
Mast cell & Microglia triggers
PI3K
AKT
mTORC1
PTEN
TSC1
TSC2
S6K
4E-BP1
cell number
ASD
cell activation
Rapamycin
Luteolin
Calcium influx via receptor
Cell size & proliferation

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4/22/2017
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4/22/2017
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4/22/2017
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4/22/2017
Copyright Dr. T.C. Theoharides

46. Structures of quercetin and luteolin
Flavonoids are lipophilic (do not dissolve in water)
Less than 5% are absorbed orally
Are heavily metabolized (broken down in the liver)
Need to formulate with oil in liposomes
Need to use in proper combination to permit some action in GI and some in brain
4/22/2017
Copyright Dr. T.C. Theoharides

47. Beneficial Effect of Flavonoids in Brain Inflammation
Middleton E Jr, Kandaswami C, Theoharides TC.
Pharmacol Rev (4): Review.
Flavonoids are potent:
Anti-oxidant and
Anti-inflammatory
Mast cell inhibitors
Metal chelators
Inhibitory Effect of Luteolin on Pro-Inflammatory
Mediator Release from Human Mast Cells
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Dec. 20, 2009
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Theoharides et al. Expert Opin. Pharmacotherapy (2009) 10 (13):
Copyright Dr. T.C. Theoharides

55. Cochrane Database Syst Rev.
2010 Aug 4;(8):CD

56. In a recent study of spontaneous reporting of adverse drug - drug interaction in Italy, the incidence was 9.8% for two drugs but increased to 88.3% for 8 drugs or more there is less information on drug-supplement interaction. However, it was reported that 33.4% of patients using antipsychotics also took traditional Chinese medicine concurrently; 7.2% of these patients had worse outcomes as compared to 4.4% of those using antipsychotic alone.

57. Allergic, environmental, immune, infectious, Stress and toxic triggers
(Foods, IL-1, LPS, CRH, mercury, PCBs)
Mast cell
Blood vessel
Luteolin
Quercetin
Mast cell-derived inflammatory, neurotoxic and oxidative stress mediators
(IL-6, TNF, tryptase)
Rutin
Disruption of the
gut-blood barrier
Disruption of the blood-brain barrier
GOAL
Luteolin formulation to increase
absorption and brain levels
Stimulation
Brain Inflammation
Inhibition

58. Flavonoids are poorly absorbed
Flavonoids are lipophilic (do not dissolve in water)
Less than 5% are absorbed orally
Are heavily metabolized (broken down in the liver)
Need to formulate with olive kernel oil
Need to use in proper combination to permit actions in both GI and in brain

60. Relevant TCT Patents and Trademark:
US patents No. 6,624,148; 6,689,748; 6,984,667, and EPO , which cover methods and compositions of mast cell blockers, including flavonoids, US patents 7,906,153 and 12/861,152 (allowed) for treatment of neuro-inflammatory conditions, as well as US patent applications No.12/534,571 and No.13/009,282 for the diagnosis and treatment of autism.
Trademark NeuroProtek® US 3,225,924

61. Benefits of NeuroProtek®
Properties
Benefits
Luteolin
Anti-oxidant
Reduces brain oxidative stress
Anti-inflammatory
Reduces brain inflammation
Microglia inhibitor
Anti-cytokine
Inhibits IL-6 production
Stabilizes blood vessels
Protects against BBB disruption
Is neuroprotective
Protects neuronal damage
Reverses autistic-like behavior in mice
Has beneficial effect in vitro
Olive Kernel Oil
Reduces gut and brain inflammation
Contains hydroxytyrosol and oleocanthal
Inhibit beta-amyloid plaque formation
Reverses Alzheimer’s condition in mice

64. Approximate improvement of children
Benefits of Neuroprotek in autism*
Symptoms
Approximate improvement of children
Stool color, shape, smell
28/37
“Allergic problems”
19/37
Eye contact
15/37
Retention of learned aspects
Social interaction
14/37
Attention to directions
12/37
Speaking words/sentences
4/37
Hyperactivity/Aggression -
* (n= 37, 4-14 years old; 29 boys and 8 girls)

65. Phenolic groups of quercetin, luteolin and Pycnogenol
Some ASD children
cannot metabolize
Polyphenolic
Compounds leading to:
Hyperactivity
Irritability
Aggressiveness
Inability to sleep
Inappropriate laughter
Self-injurious behavior
Test-Administer teaspoon of acetaminophen (Tylenol)
Reduce-Administer NoFenol (Houston Enzymes)
Pycnogenol has 15, quercetin has 5
and luteolin 4 phenolic groups

66. Comparison of Luteolin and Methoxyluteolin on Inhibition of Human Mast Cell TNF Release

67. Sofia’s Progress Dx: Autistic disorder 11/2010-Non verbal
Started on NeuroProtek:
6/2011-”Sentences, singing and drawing”

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Christina’s Progress
Dx: Autistic disorder
3/2011-Nonverbal
Started on NeuroProtek:
4/2011-”Can I play with you?”
4/22/2017
Copyright Dr. T.C. Theoharides

70. Mack’s Progress Dx: Autistic disorder 11/2010-Non verbal
Started on NeuroProtek:
12/2011-”Xmas carrols, drawing first Xmas card ever,”
6/2011-”Sentences, singing
1/2012-”Full conversation, attending regular middle school.”

71. http://www.mastcellmaster.com http://www.neuroprotek.com
Research Funding:
DK62861 NIDDK AR47652 NIAMS NS NINDS NS55681 NINDS AR60951 NIAMS
NS66205 NIH
Autism Research Collaborative
Safe Minds
Autism Research Institute

72. Portion of any profits will be donated to autism research
Disclosure
Dr. Theoharides is Medical Director
and holds a major interest in Algonot, LLC
Portion of any profits will be donated to autism research