AU2019308501B2 - Compositions and methods for treating autism - Google Patents

Abstract

The invention provides compositions and methods of treating autism. Specifically, the invention relates to treating the core symptoms of autism by administering alpha-methyl-DL-tyrosine.

Description

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COMPOSITIONS AND METHODS FOR TREATING AUTISM CROSS-REFERENCE TO RELATED APPLICATIONS

[001] This application claims priority to and the benefit of United States Patent Application

16/040,405, filed July 19, 2018, which is incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

[002] The invention relates to compositions and methods of treating autism. Specifically, the

invention relates to treating the core symptoms of autism by administering a-methyl-DL-tyrosine. -methyl-DL-tyrosine.

BACKGROUND OF THE INVENTION

[003] Autism spectrum disorder (ASD) or autism is defined in the Diagnostic and Statistics

Manual of Mental Disorders V (DSM-5) by "difficulties in social communication and social

interaction, and restricted and repetitive behavior, interests or activities". Childhood autism is

more prevalent than childhood cancer, juvenile diabetes, and pediatric acquired immunodeficiency

syndrome (AIDS) combined, with an estimated prevalence of 1.5 million in the United States (US),

3 million children in Europe and tens of millions throughout the rest of the world. Autism also

represents a substantial economic burden in both children and adults. More disturbing is that, for

no explicable reason, childhood autism appears to be increasing at a rate of 10-17% per year.

Lastly, there are currently no approved medications that address the core symptoms of autism.

Autism is a serious disease that represents an area of significant economic burden and unmet

medical need.

[004] Autism was first described in 1943 by Dr. Leo Kanner who described his 5 year old patient

as, as, "" '...happiest whenleft happiest when left alone, alone, almost almostnever cried never to go cried to with his mother, go with did notdid his mother, seemnot to seem noticeto notice

his father's homecomings, and was indifferent to visiting relatives wandered about smiling,

making stereotyped movements with his fingers spun with great pleasure anything he could seize

upon to spin Words to him had a specifically literal, inflexible meaning When taken When taken into ainto a

room, he completely disregarded the people and instantly went for objects", as cited by Lai, M.C.,

et al. Autism. Lancet, 2014. 383(9920): p. 896-910.

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[005] Originally autism was thought to be a form of childhood schizophrenia. However, in the

mid-1980s, mid-1980s, it it was was found found that that autism autism is is aa heritable heritable disorder disorder and and believed believed to to have have aa genetic genetic etiology. etiology.

Currently, autism is defined diagnostically in the Diagnostic and Statistical Manual of Mental

Disorders, 5th Edition 5 Edition (DSM-5) (DSM-5) (2013) (2013) byby "difficulties "difficulties inin social social communication communication and and social social

interaction, and restricted and repetitive behavior, interests or activities." The diagnostic criteria

in DSM-5 for the features "difficulties in social communication and social interaction" of ASD

include (1) deficits in social-emotional reciprocity, ranging, for example, from abnormal social

approach and failure of normal back-and-forth conversation; to reduced sharing of interests,

emotions, or affect; to failure to initiate or respond to social interactions; (2) Deficits in nonverbal

communicative behaviors used for social interaction, ranging, for example, from poorly integrated

verbal and nonverbal communication; to abnormalities in eye contact and body language or deficits

in understanding and use of gestures; to a total lack of facial expressions and nonverbal

communication; and (3) deficits in developing, maintaining, and understanding relationships,

ranging, for example, from difficulties adjusting behavior to suit various social contexts; to

difficulties in sharing imaginative play or in making friends; to absence of interest in peers.

[006] Typically displayed in early childhood, autism is associated with many co-morbidities that

include epilepsy, Fragile-X syndrome, Retts syndrome, attention deficit/hyperactivity disorder

(ADHD), abnormal sensory or motor responses, disturbed sleep, reduced cognitive functionality,

anxiety and aggression.

[007] In 2010, the CDC reported the rate of autism in the US to be 1 in 68 children, with boys

being 5 times more susceptible than girls. This means that 1 in 42 boys are diagnosed with autism.

There was an increase of about 30% since the assessment of autism prevalence conducted in 2014,

and more than double the rate that was reported only 12 years and the problem is growing rapidly.

In New Jersey the observed rate of autism was 1 in 46 children, which means that 1 in 29 boys

born in New Jersey are likely to be autistic.

[008] The financial cost of raising an autistic child was estimated in 2014 to be $3.2M more than

the cost of raising a non-autistic child, and this does not take into account the societal costs of

maintaining this population as adults once their families are no longer able to do SO. so. The societal

costs of autism are broad and deep, and many have never been explored. For example, it was only

in mid-2017 that information was developed on the rate of healthcare utilization by autistics and it was found that their need for psychiatric care as well as care for the high incidence of autism associated comorbidities was far beyond that of the general population or other elements of the psychiatric patient population. Similarly, it was not until September of 2017 that the rate of school suspension and expulsion was dramatically higher in the autistic population, and growing as the autistic population grew in numbers.

[009] Recently, attention has been brought to bear on autism associated mortality rates. Although

autism is not typically considered to be a fatal disease, a number of investigators have reported a

significantly increased mortality in the autistic population with the major cause of death being

suicide. A matched case cohort study based upon the Swedish National Patient Registry and the

Cause of Death Registry looked at deaths between 1987 and 2009, and found a 256% greater death

rate in autistic patients compared to the general population. The mean age at the time of death was

70.2 years for the general population and 58.39 for patients with autism, with suicide associated

with better performing patients. A review of 1,706 children and adolescents reported an 18%

increased risk of suicidal ideation or attempts in autism. 35% of patients with Asperger's

syndrome were reported in a Canadian study to have attempted suicide. Similarly, in Japan,

Australia, England, and Belgium. In a French review of the PubMed literature, it was found that

overall 21.3% of autism patients reported suicidal ideation or had attempted suicide, with the

noteworthy noteworthyobservation thatthat observation " ... " the the methods methodsused areare used often violent". often violent".

[010] Enormous resources are being spent on research into the causes of autism. Genetic, dietary,

developmental, pharmacologic, environmental, and behavioral elements have all been implicated

as potential causes of this syndrome.

[011] A purified, single enantiomer presentation of molecule - a-methyl-L-tyrosine, currently -methyl-L-tyrosine, currently

marketed as Demser®, is known to inhibit tyrosine hydroxylase, which is the rate limiting enzyme

in the biosynthesis of catecholamine's consequent to enzyme-mediated conversion of L-tyrosine

to DOPA (dihydroxy-phenylalanine). This first step in catecholamine synthesis is highly

stereospecific for L-tyrosine. L-AMPT, a methylated L-tyrosine, acts as a competitive inhibitor

for tyrosine hydroxylase (Demser (Demser®Prescribing PrescribingInformation, Information,2015; 2015;FDA FDASummary SummaryBasis Basisof of

Approval [SBA]for Approval [SBA] for Demser1979). Demser®, 1979).Demser® Demserwas was initially initially approved approved byFDA by the theinFDA inand 1979 1979 and

is indicated for use in the treatment of patients with pheochromocytoma for (1) the preoperative

preparation of patients for surgery, (2) management of patients when surgery is contraindicated,

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and (3) chronic treatment of patients with malignant pheochromocytoma. The recommended

initial dosage of Demser Demser®for foradults adultsand andchildren children12 12years yearsof ofage ageand andolder olderis is250 250mg mgorally orallyfour four

times daily. This dosage may be increased by 250 mg to 500 mg every day to a maximum of 4.0

g/day in 4 divided doses. Although Demser Demser®is iscommercially commerciallyavailable, available,those thoseordinarily ordinarilyskilled skilled

in the art have not developed methods for treating autism or its core symptoms by administering a

racemic mixture of a-methyl-DL-tyrosine, i.e., DL-AMPT. -methyl-DL-tyrosine, i.e., DL-AMPT.

[012] Given the widespread concern over (a) dramatic increases in prevalence of autism, (b)

devastating impact of the disorder on children and their families, and (c) substantial social and

economic burden that autism place on communities, educational, medical and mental health

systems within our society, autism represents a public health emergency. Accordingly, there exists

a vital need for therapeutically effective methods for treating autism, in particular, the core

symptoms of this disorder.

SUMMARY OF THE INVENTION

[013] In one aspect, the invention provides a method for treating an autism in a subject in need

thereof, the method comprising administering to said subject a composition comprising a

therapeutically effective amount of a-methyl-DL-tyrosine andaapharmaceutically -methyl-DL-tyrosine and pharmaceuticallyacceptable acceptable

carrier. In an exemplary embodiment, the composition comprises a-methyl-DL-tyrosine in an -methyl-DL-tyrosine in an

amount ranging from about 50 mg (w/w) to about 500 mg (w/w).

[014] In another aspect, the invention provides a method for providing a plasma concentration of

a therapeutic drug for a long term for treating an autism in a subject in need thereof, the method

comprising administering to said subject said therapeutic drug at a concentration ranging from

about 50 mg (w/w) to about 500 mg (w/w) three times a day, wherein said therapeutic drug is a- -

methyl-DL-tyrosine, wherein said plasma concentration ranges from about 500 ng/ml to 5000

ng/ml, and wherein said term is at least 1 week.

[015] In another aspect, the invention provides a method for treating an autism associated clinical

trait in a subject in need thereof, the method comprising administering to said subject a

composition comprising a therapeutically effective amount of a-methyl-DL-tyrosine, thereby -methyl-DL-tyrosine, thereby

treating said autism associated clinical trait in said subject. In an exemplary embodiment, the

clinical trait is a deficit in social communication, a deficit in social interaction, a deficit in social

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motivation, lethargy and social withdrawal, inappropriate speech, hyperactivity, stereotypic

behavior, irritability and agitation, restrictive behavior, repetitive behavior, ritualistic behavior,

sameness behavior, compulsive behavior, self-injurious behavior or a combination thereof.

[016] Other features and advantages of the present invention will become apparent from the

following detailed description examples and figures. It should be understood, however, that the

detailed description and the specific examples while indicating preferred embodiments of the

invention are given by way of illustration only, since various changes and modifications within

the spirit and scope of the invention will become apparent to those skilled in the art from this

detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

[017] Figure 1 depicts Conners Parent Rating Scale Data results.

[018] Figure 2 illustrates patient enrollment and disposition.

[019] Figure 3 shows an example of largest change from baseline in vital signs noted in a patient

in Study HT 02-121 (Example 6) and demonstrates that these changes were transient, falling far

short of orthostatic criteria.

[020] Figure 4 shows random L1-79 plasma concentrations (except Day 1*) by week, overall

mean and by patient from open-label 100 mg and 200 mg TID in Study HT 02-121. *1 On Day 1,

L-79 plasma concentrations were evaluated at 1-hour post-dose.

[021] Figure 5 shows random L1-79 plasma concentrations by week (excluding Day 1), overall

mean and by patient from open-label 100 mg TID group in Study HT 02-121.

[022] Figure 6 shows random L1-79 plasma concentrations by week (excluding Day 1), overall

Mean and by patient from open-label 200 mg TID group in Study HT 02-121.

[023] Figure 7 graphically displays shows the CGI - Overall Severity (CGI-S) for L1-79

compared to placebo over time.

[024] Figure 8 graphically displays the CGI - -Overall Improvement (CGI-I) Overall Improvement (CGI-I) for for L1-79 L1-79 compared compared

to to placebo placeboover time. over time.

[025] Figure 9 shows the change from baseline in CGI-S at week 4 (ITT Population, Draft).

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[026] Figure 10 shows the change in CGI-S from Baseline to Week 4 by Patient (ITT Population,

Draft).

[027] Figure 11 shows the responder analysis for CGI-S at Week 4/LOCF (ITT Population,

Draft).

[028] Figure 12 shows the change from Baseline in VABS II Standardized Socialization Score,

Week 0 - Week 8 (ITT Population, Draft).

[029] Figure 13 shows the change from Baseline in VABS II Standardized Communication

Score, Week 0 - Week 8 (ITT Population, Draft).

[030] Figure 14 shows the change from Screening in ADOS-2 Total score, Week 0 - Week 4

(ITT Population, Draft).

[031] Figure 15 shows the change from Screening in ADOS-2 Restrictive and Repetitive

Behavior Total score, Week 0 - Week 4 (ITT Population, Draft).

[032] Figure 16 shows the change from Screening in ADOS-2 Social Affect Total Score, Week

0 - Week 4 (ITT Population, Draft).

[033] Figure 17 shows the change in ADOS-2 Total Score from Screening to Week 4 by Patient

(ITT (ITT Population, Population,Draft). Draft).

[034] Figure 18 shows the change from Baseline in SRS-2 Total T-score, Week 0 - Week 8 (ITT

Population, Draft).

[035] Figure 19 shows the change from Baseline in SRS-2 DSM-5 Social Communication and

Interaction T-score, Week 0 - Week 8 (ITT Population, Draft).

[036] Figure 20 shows the change from Baseline in SRS-2 Social Communication T-score, Week

0 - Week 8 (ITT Population, Draft).

[037] Figure 21 shows the change from Baseline in SRS-2 Social Motivation T-score, Week 0 -

Week 8 (ITT Population, Draft).

[038] Figure 22 shows the change from Baseline in SRS-2 DSM-5 Restrictive and Repetitive

Behavior, Week 0 - Week 8 (ITT Population, Draft).

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[039] Figure 23 shows the change in SRS-2 Total T-score from Baseline to Week 4 by Patient

(ITT (ITT Population, Population,Draft). Draft).

[040] Figure 24 shows the change in SRS-2 DSM-5 Social Communication and Interaction T-

score from Baseline to Week 4 by Patient (ITT Population, Draft).

[041] Figure 25 shows the change in SRS-2 Social Communication T-score from Baseline to

Week 4 by Patient (ITT Population, Draft).

[042] Figure 26 shows the change in SRS-2 Social Motivation T-score from Baseline to Week

4 by Patient (ITT Population, Draft).

[043] Figure 27 shows the responder analysis for SRS-2 Total T-score at Week 4/LOCF (ITT

Population, Draft).

[044] Figure 28 shows the responder analysis for SRS-2 DSM-5 Social Communication and

Interaction T-score at Week 4/LOCF (ITT Population, Draft).

[045] Figure 29 shows the responder analysis for SRS-2 Social Communication T-score at Week

4/LOCF (ITT Population, Draft).

[046] Figure 30 shows the responder analysis for SRS-2 Social Motivation T-score at Week

4/LOCF (ITT Population, Draft).

[047] Figure 31 shows the change from Baseline in ABC-C Lethargy and Social Withdrawal

Domain, Week 0 - Week 8 (ITT Population, Draft).

[048] Figure 32 shows the change from Baseline in ABC-C Inappropriate Speech Domain, Week

0 - Week 8 (ITT Population, Draft).

[049] Figure 33 shows the change from Baseline in ABC-C Hyperactivity and Noncompliance

Domain, Week 0 - Week 8 (ITT Population, Draft).

[050] Figure 34 shows the change from Baseline in ABC-C Stereotypic Behavior Domain, Week

0 - Week 8 (ITT Population, Draft).

[051] Figure 35 shows the change from Baseline in ABC-C Irritability and Agitation Domain,

Week Week 00 -- Week Week 88 (ITT (ITT Population, Population, Draft). Draft).

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[052] Figure 36 shows the change in ABC-C Lethargy and Social Withdrawal Domain from

Baseline to Week 4 by Patient (ITT Population, Draft).

[053] Figure 37 shows the change in ABC-C Inappropriate Speech Domain from Baseline to

Week 4 by Patient (ITT Population, Draft).

[054] Figure 38 shows the responder analysis for ABC-C Lethargy and Social Withdrawal

Domain at Week 4/LOCF (ITT Population, Draft).

[055] Figure 39 shows the responder Analysis for ABC-C Inappropriate Speech Domain at

Week 4/LOCF (ITT Population, Draft).

[056] Figure 40 shows the change from Baseline in RBS-R Total Score, Week 0 - Week 8 (ITT

Population, Draft).

[057] Figure 41 shows the change from Baseline in RBS-R Restrictive Behavior, Week 0 - Week

8 (ITT Population, Draft). There were mean improvements in RBS-R Total Score as noted by a

decrease in score for L1-79 200 mg compared to placebo.

[058] Figure 42 shows the change from Baseline in RBS-R Ritualistic Behavior, Week 0 -Week

8 (ITT Population, Draft).

[059] Figure 43 shows the change from Baseline in RBS-R Sameness Behavior, Week 0 - Week

8 (ITT Population, Draft).

[060] Figure 44 shows the change from Baseline in RBS-R Compulsive Behavior, Week 0 -

Week 8 (ITT Population, Draft).

[061] Figure 45 shows the change from Baseline in RBS-R Stereotypic Behavior, Week 0 -

Week 8 (ITT Population, Draft).

[062] Figure 46 shows the change from Baseline in RBS-R Self-injurious Behavior, Week 0 -

Week 8 (ITT Population, Draft).

[063] Figure 47 shows the change in RBS-R Total Score from Baseline to Week 4 by Patient

(ITT Population, Draft)

[064] Figure 48 shows the responder analyses (defined as an improvement or no worsening) for

RBS-R Total Score at Week 4 or LOCF.

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DETAILED DESCRIPTION OF THE INVENTION

[065] The present subject matter may be understood more readily by reference to the following

detailed description which forms a part of this disclosure. It is to be understood that this invention

is not limited to the specific products, methods, conditions or parameters described and/or shown

herein, and that the terminology used herein is for the purpose of describing particular

embodiments by way of example only and is not intended to be limiting of the claimed invention.

[066] Unless otherwise defined herein, scientific and technical terms used in connection with the

present application shall have the meanings that are commonly understood by those of ordinary

skill in the art. Further, unless otherwise required by context, singular terms shall include

pluralities and plural terms shall include the singular.

[067] As employed above and throughout the disclosure, the following terms and abbreviations,

unless otherwise indicated, shall be understood to have the following meanings.

[068] In the present disclosure the singular forms "a," "an," and "the" include the plural reference,

and reference to a particular numerical value includes at least that particular value, unless the

context clearly indicates otherwise. Thus, for example, a reference to "a compound" is a reference

to one or more of such compounds and equivalents thereof known to those skilled in the art, and

SO so forth. The term "plurality", as used herein, means more than one. When a range of values is

expressed, another embodiment includes from the one particular and/or to the other particular

value. Similarly, when values are expressed as approximations, by use of the antecedent "about,"

it is understood that the particular value forms another embodiment. All ranges are inclusive and

combinable.

[069] As used herein, the terms "component," "composition," "composition of compounds,"

"compound," "drug," "pharmacologically active agent," "active agent," "therapeutic," "therapy,"

"treatment," or "medicament" are used interchangeably herein to refer to a compound or

compounds or composition of matter which, when administered to a subject (human or animal)

induces a desired pharmacological and/or physiologic effect by local and/or systemic action.

[070] As used herein, the ensuing terms are defined, as follows:

Applied Behavioral Analysis ABA Aberrant Behavior Checklist - Community ABC-C

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Attention Deficit Hyperactivity Disorder ADHD Autistic Diagnosis Interview Review ADIR Absorption, Distribution, Metabolism and Elimination ADME ADOS-2 Autism Diagnosis Observation Schedule, Second Edition

Adverse Events AE

AIDS Acquired immunodeficiency syndrome

Autism spectrum disorder ASD ASD Aspartate Aminotransferase AST Alanine Aminotransferase ALT Area under the concentration time curve AUC AUC AUC0-8 Area under the concentration time curve from 0 to 8 hours AUC- AUC0-12 Area under the concentration time curve from 0 to 12 hours AUC- AUC0-00 Area under the concentration time curve from 0 to infinity AUC- AUCO-tz Area under the concentration time curve from 0 to the last quantifiable AUC- concentration

Code of Federal Regulations CFR CGI-I Clinical Global Impression - Overall Improvement

CGI-S Clinical Global Impression - Severity of Illness

Cmax Maximum Concentration

Central Nervous System CNS Racemic (D and L isomer) a-methyl-para-tyrosine -methyl-para-tyrosine DL-AMPT Dihydroxy-phenylalanine DOPA DSM-5 Diagnostic and Statistics Manual of Mental Disorders V

EI Early Intervention

EOP2 End of Phase 2 EOP2 Food and Drug Administration FDA FDASIA Food and Drug Administration Safety and Innovation Act FDASIA

FOBs Functional Observation Battery Assessments

Good Manufacturing Practice GMP Hepatitis B Virus HBV HBV Hemorrhagic Cerebrovascular Accident HCVA Human Equivalent Doses HED HIV Human Immunodeficiency Virus

IEP IEP Individualized Educational Program

L isomer of a-methyl-para-tyrosine -methyl-para-tyrosine L-AMPT

LC-MS/MS Liquid Chromatographic Mass Spectrometric

Last Observation Carried Forward LOCF

LQTS Long QT Syndrome

Maximum Tolerated Dose MTD New Drug Application NDA No Observed Adverse Effect Level NOAEL PD Pharmacodynamic

Pharmacokinetic PK PK Post Natal Day PND PSP PSP Pediatric Study Plan

RBS-R Repetitive Behavior Score - Revised

SBA Summary Basis of Approval

SRS-2 Social Responsiveness Scale, Second Edition

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t1/2 Half Life t/

TdP Torsade's de Pointes

TID Three times daily

Toxicokinetic TK TQT Thorough QT

Upper Limit of Normal ULN United States Adopted Name USAN VABS II Vineland Adaptive Behavior Scales, Second Edition

[071] With respect to autism, for example, the negative effect or symptoms can include any of

those those that that are are the the subject subject of of the the diagnostic diagnostic criteria criteria specified specified for for autism autism spectrum spectrum disorder disorder in in the the

American Psychiatric Association's Diagnostic and Statistical Manual, Fifth Edition (DSM-5,

DSM-V), the contents of which are incorporated by reference herein, e.g., deficits in social-

emotional reciprocity (ranging, for example, from abnormal social approach and failure of normal

back-and-forth conversation; to reduced sharing of interests, emotions, or affect; to failure to

initiate initiate or or respond respond to to social social interactions) interactions) deficits deficits in in nonverbal nonverbal communicative communicative behaviors behaviors used used for for

social interaction (ranging, for example, from poorly integrated verbal and nonverbal

communication; to abnormalities in eye contact and body language or deficits in understanding

and use of gestures; to a total lack of facial expressions and nonverbal communication); deficits in

developing, developing, maintaining, maintaining, and and understanding understanding relationships relationships (ranging, (ranging, for for example, example, from from difficulties difficulties

adjusting behavior to suit various social contexts; to difficulties in sharing imaginative play or in

making friends; to absence of interest in peers); stereotyped or repetitive motor movements, use

of of objects, objects, or or speech speech (e.g., (e.g., simple simple motor motor stereotypes, stereotypes, lining lining up up toys toys or or flipping flipping objects, objects, echolalia, echolalia,

idiosyncratic phrases); insistence on sameness, inflexible adherence to routines, or ritualized

patterns of verbal or nonverbal behavior (e.g., extreme distress at small changes, difficulties with

transitions, rigid thinking patterns, greeting rituals, need to take same route or eat same food every

day); highly restricted, fixated interests that are abnormal in intensity or focus (e.g., strong

attachment to or preoccupation with unusual objects, excessively circumscribed or perseverative

interests); and hyper- or hyporeactivity to sensory input or unusual interest in sensory aspects of

the environment (e.g. apparent indifference to pain/temperature, adverse response to specific

PCT/US2019/040581

sounds or textures, excessive smelling or touching of objects, visual fascination with lights or

movement).

[072] Some subpopulations of patients on the autism spectrum, include those patients diagnosed

with Asperger's Disorder (i.e., Asperger Syndrome) or Social Communication Disorder, exhibit

symptoms of Attention Deficit Hyperactivity Disorder (ADHD) (e.g., inattention, hyperactivity,

and impulsivity) and/or tics (motor tics or vocal tics). See, e.g., DSM-5.

[073] Assessment of autism symptoms, or any of the symptoms of the present disclosure, can be

performed using methods known in the art. For example, one method of assessing autism

symptoms is the Clinical Global Impressions (CGI) scale based upon changes from baseline in

various psychometric tests. These tests can include the Aberrant Behavior Checklist-Community

(ABC-C), Conners Parent Rating Scale and the Autism Diagnostic Observation Schedule (ADOS).

Autism symptoms can also be assessed from the clinician's personal, clinical observations, from

videographic videographic information information taken taken at at regularly regularly scheduled scheduled clinic clinic visits, visits, and and from from information information provided provided

by the subject's caregivers over time. Compositions and methods of the disclosure result in a

reduction of at least 1 point in at least one dimension of the Conners Parent Rating Scale

assessment score.

[074] As employed above and throughout the disclosure the term "effective amount" refers to an

amount effective, at dosages, and for periods of time necessary, to achieve the desired result with

respect to the treatment of the relevant disorder, condition, or side effect. It will be appreciated

that the effective amount of components of the present invention will vary from patient to patient

not only with respect to the particular compound, component or composition selected, the route of

administration, and the ability of the components to elicit a desired result in the individual, but

also with respect to factors such as the disease state or severity of the condition to be alleviated,

hormone levels, age, sex, weight of the individual, the state of being of the patient, and the severity

of the pathological condition being treated, concurrent medication or special diets then being

followed by the particular patient, and other factors which those skilled in the art will recognize,

with the appropriate dosage being at the discretion of the attending physician. Dosage regimes may

be adjusted to provide improved therapeutic response. An effective amount is also one in which

any toxic or detrimental effects of the components are outweighed by the therapeutically beneficial

effects.

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[075] The invention also provides a pharmaceutical composition comprising compounds of the

invention and one or more pharmaceutically acceptable carriers. "Pharmaceutically acceptable

carriers" include any excipient which is nontoxic to the cell or mammal being exposed thereto at

the dosages and concentrations employed. The pharmaceutical composition may include one or

additional therapeutic agents.

[076] "Pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or

dosage forms which are, within the scope of sound medical judgment, suitable for contact with the

tissues of human beings and animals without excessive toxicity, irritation, allergic response, or

other problem complications commensurate with a reasonable benefit/risk ratio.

[077] Examples of a pharmaceutically acceptable carrier include, for example, but not limited to,

solvents, dispersion media, buffers, coatings, antibacterial and antifungal agents, wetting agents,

preservatives, buggers, chelating agents, antioxidants, isotonic agents and absorption delaying

agents.

[078] Examples of a pharmaceutically acceptable carrier also include, for example, but not

limited to, water; saline; phosphate buffered saline; dextrose; glycerol; alcohols such as ethanol

and isopropanol; phosphate, citrate and other organic acids; ascorbic acid; low molecular weight

(less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or

immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as

glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other

carbohydrates including glucose, mannose, or dextrins; EDTA; salt forming counterions such as

sodium; and/or nonionic surfactants such as TWEEN, polyethylene glycol (PEG), and

PLURONICS; isotonic agents such as sugars, polyalcohols such as mannitol and sorbitol, and

sodium chloride; as well as combinations thereof.

[079] Within the present invention, the disclosed compounds may be prepared in the form of

pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" refer to derivatives of the

disclosed compounds wherein the parent compound is modified by making acid or base salts

thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or

organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such

as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional

non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example,

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from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include

those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic,

phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic,

succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,

phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic,

methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like. These physiologically

acceptable salts are prepared by methods known in the art, e.g., by dissolving the free amine bases

with an excess of the acid in aqueous alcohol, or neutralizing a free carboxylic acid with an alkali

metal base such as a hydroxide, or with an amine.

[080] Compounds described herein can be prepared in alternate forms. For example, many

amino-containing compounds can be used or prepared as an acid addition salt. Often such salts

improve isolation and handling properties of the compound. For example, depending on the

reagents, reaction conditions and the like, compounds as described herein can be used or prepared,

for example, as their hydrochloride or tosylate salts. Isomorphic crystalline forms, all chiral and

racemic forms, N-oxide, hydrates, solvates, and acid salt hydrates, are also contemplated to be

within the scope of the present invention.

[081] Certain acidic or basic compounds of the present invention may exist as zwitterions. All

forms of the compounds, including free acid, free base and zwitterions, are contemplated to be

within the scope of the present invention. It is well known in the art that compounds containing

both amino and carboxy groups often exist in equilibrium with their zwitterionic forms. Thus, any

of the compounds described herein that contain, for example, both amino and carboxy groups, also

include reference to their corresponding zwitterions.

[082] The term "stereoisomers" refers to compounds that have identical chemical constitution,

but differ as regards the arrangement of the atoms or groups in space. The term "enantiomers"

refers to stereoisomers that are mirror images of each other that are non-superimposable.

[083] The term "administering" means either directly administering a compound or composition

of the present invention, or administering a prodrug, derivative or analog which will form an

equivalent amount of the active compound or substance within the body.

[084] The term "inhibitor" as used herein includes compounds that inhibit the expression or

activity of a protein, polypeptide or enzyme and does not necessarily mean complete inhibition of expression and/or activity. Rather, the inhibition includes inhibition of the expression and/or activity of a protein, polypeptide or enzyme to an extent, and for a time, sufficient to produce the desired effect.

[085] While not intending to be bound by any particular mechanism of operation, it is believed

that the tyrosine hydroxylase inhibitors according to the present invention function by decreasing

the amount of adrenaline secreted into the bloodstream.

[086] The tyrosine hydroxylase inhibitor is well known in the art and fully described in, for

example, U.S. Patent Application Publications US 2015/0290279, US 2015/0216827, US

2015/0111937, US 2015/0111878, US 2013/0184214, and US 20130183263; U.S. Patents US

8,481,498, US 9,308,188, and US 9,326,962; and PCT Patent Application Publication

WO2015061328, which are incorporated by reference herein in their entirety. Any suitable

tyrosine hydroxylase inhibitor, known to one of skilled in the art, can be used.

[087] In certain embodiments, the tyrosine hydroxylase inhibitor is a tyrosine derivative. The

tyrosine derivative can be capable of existing in different isomeric forms, including stereoisomers

and enantiomers. The tyrosine derivative can, for example, exist in both L-form or D-form. The

tyrosine derivative can, for example, also exist in a racemic form.

[088] Representative tyrosine derivatives include, for example, one or more of methyl (2R)-2-

amino-3-(2-chloro-4 hydroxyphenyl) propanoate, D-tyrosine ethyl ester hydrochloride, methyl

(2R)-2- amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoate H-D-tyrosine(tBu)-allyl ester

(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, hydrochloride, methyl 1(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate,methyl methyl(2R)- (2R)-

2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl (2R)-2-amino-3-(4-[(2-

chloro-6-fluorophenyl) methoxy] phenyl) propanoate, methyl (2R)-2- amino-3-(2-chloro-3,4-

dimethoxyphenyl) propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)

propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy] benzyl malonate,

methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl) propanoate, methyl (2R)-2-amino-3-(3-

chloro-4-hydroxy-5-methoxyphenyl) propanoate, methyl (2R)-2-amino-3-(2,6- dichloro-3-

(2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) hydroxy-4-methoxyphenyl) propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxypheryl)

propanoate, H-DL-tyrosine methyl ester hydrochloride, H-3,5-diiodo-tyrosine methyl ester

hydrochloride, H-D-3,5-diiodo-tyrosine methyl ester hydrochloride, H-D-tyrosine methyl ester

hydrochloride, D-tyrosine methyl ester hydrochloride, D-tyrosine-methyl ester hydrochloride, wo 2020/018292 WO PCT/US2019/040581 methyl D-tyrosinate hydrochloride, H-D-tyrosine methyl ester hydrochloride, D-tyrosine methyl ester ester hydrochloride,H-D-tyrosine hydrochloride, H-D-tyrosinemethyl methylester-hydrochloride, ester-hydrochloride,(2R)-2-amino-3-(4- (2R)-2-amino-3-(4- hydroxyphenyl) propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl ester hydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoate hydrochloride, methyl (2R)-2-azanyl-3-(4- hydroxyphenyl) propanoate hydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine, 3-nitro-L- tyrosine ethyl ester hydrochloride, DL-m-tyrosine, DL-o-tyrosine, Boc-tyrosine (3,5-I2)-OSu, (3,5-12)-OSu,

Fmoc-tyrosine(3-N02)-OH, a-methyl-L-tyrosine, a-methyl-D-tyrosine, -methyl-L-tyrosine, -methyl-D-tyrosine, and and a-methyl-DL- -methyl-DL-

tyrosine. In certain embodiments of the invention, the tyrosine derivative is a-methyl-L-tyrosine -methyl-L-tyrosine

as shown below:

O OH NH2 HO NH

[089] In other embodiments, the tyrosine derivative is a-methyl-D-tyrosine. Inother -methyl-D-tyrosine. In other

embodiments, the tyrosine derivative is a-methyl-DL-tyrosine in aa racemic -methyl-DL-tyrosine in racemic form form as as shown shown below: below:

O OH H3C NH2 HC NH HO

[090] In a particular embodiment, the tyrosine derivative is a structural variant of a-methyl-L- -methyl-L-

tyrosine or a-methyl-DL-tyrosine. Thestructural -methyl-DL-tyrosine. The structuralvariants variantsof of-methyl-L-tyrosine a-methyl-L-tyrosine oror a-methyl- -methyl-

DL-tyrosine are well known in the art and fully described in, for example, U.S. Patent 4, 160,835,

which is incorporated by reference herein in its entirety.

WO wo 2020/018292 PCT/US2019/040581

[091] In one embodiment, the tyrosine derivative of the invention is an arylalanine compound

having the formula:

R1 R 0 R3-CH2 c c R CH o R2 N O R H R4

wherein whereinR1R is is hydrogen, hydrogen,methyl or ethyl methyl esterester or ethyl group,group, or alkyl or of from of alkyl 1 tofrom 4 carbon 1 to atoms; 4 carbon atoms;

R2 is hydrogen, R is hydrogen, lower lower alkyl, alkyl, lower lower alkene, alkene, succinimide, succinimide, or or alkyl alkyl of of from from 11 to to 44 carbon carbon atoms; atoms; RR3

is a substituted benzene ring of the following general formula

Y1 Y Y2 Y3 Y Y

wherein Y1, is located Y, is located at at the the para para position position and and is is hydrogen, hydrogen, hydroxy, hydroxy, methyl methyl ether, ether, dimethyl dimethyl

ether, ether, trimethyl trimethylether, or an ether, or unsubstituted or halogen-substituted an unsubstituted benzyl; Y2, or halogen-substituted and Y3 Y, benzyl; are and the Y same are the same

or or different differentand wherein and one one wherein or both Y2, and or both Y, Y3 located and at either Y located meta position at either or ortho position, meta position or ortho position,

and and wherein whereinY2, Y,and andY3Y are arehydrogen, hydroxy, hydrogen, halogen, hydroxy, methylmethyl halogen, ether, ether, or nitro; or and R4 is and nitro; hydrogen, R4 is hydrogen,

acetyl, tert-butyloxycarbonyl or fluorenylmethyloxycarbonyl.

[092] In some embodiments, Y1 and Y2 are the same or different and are selected from hydrogen,

cyanoamino, carboxyl, cyano, thiocarbamoyl, aminomethyl, guanidino, hydroxy, methanesulfonamido, nitro, amino, methanesulfonyloxy, carboxymethoxy, formyl, methoxy and

a substituted or unsubstituted 5- or 6-membered heterocyclic ring containing carbon and one or

more nitrogen, sulfur or oxygen atoms, specific examples of such heterocyclic rings being pyrrol-

1-yl, 2-carboxypyrrol-1-yl, imidazol-2-ylamino, indol-1-yl, carbazol-9-yl, 4,5-dihydro-4-

hydroxy-4-trifluoromethylthiazol-3-yl, 4-trifluoromethylthiazol-2-yl, imidazol-2-yl and 4,5-

dihydroimidazol-2-yl, such that (a) Y1 and Y2 cannot both be hydroxy, (b) Y1 and Y2 cannot both

be hydrogen and (c) when one of Y1 and Y2 is hydrogen, the other cannot be hydroxyl.

R3is

[093] In one example, R isaasubstituted substitutedor orunsubstituted unsubstitutedbenzoheterocyclic benzoheterocyclicring ringhaving havingthe the

formula:

het I

in which the benzoheterocyclic ring is selected from the group consisting of indolin-5-yl,

1-(N-benzoylcarbamimidoyl)-indolin-5-yl, 1-carbamimidoylindolin-5-yl, 1 H-2-oxindol-5-yl,

indol-5-yl, -mercaptobenzimidazol-5(6)-yl, 2-mercaptobenzimidazol-5(6)-yl, 2-aminobenzimidazol-5(6)-yl, 2-

methanesulfonamido-benzimidazol-5(6)-yl 11 H-benzoxazol-2-on-6-yl, methanesulfonamido-benzimidazol-5(6)-yl, H-benzoxazol-2-on-6-yl, 2-aminobenzothiazol-6- 2-aminobenzothiazol-6-

yl, 2-amino-4-mercaptobenzothiazol-6-yl, 2,1,3-benzothiadiazol-5-yl, 1,3-dihydro-2,2-dioxo-

1,3-dihydro-1,3-dimethyl-2,2-dixo-2,1,3-benzothiadiazol-5-yl, 4- 2,1,3-benzothiadiazol-5-yl, 1,3-dihydro-1,3-dimethyl-2,2-dioxo-2,1,3-benzothiadiazol-5-yl, 4-

methyl-2(1H)-oxoquinolin-6-yl,quinoxalin-6-yl,2-hydroxquinoxalin-6-yl,2-hydroxyquinoxalin- methyl-2(1H)-oxoquinolin-6-yl, quinoxalin-6-yl_2-hydroxquinoxalin-6-yl,2-hydroxyquinoxalin-

7-yl, 2,3-dihydroxyquinoxalin-6-yl and 2,3-dihydro-3(4H)-oxo-1,4-benzoxazin-7-yl. 2,3-dihydro-3(4 H)-oxo-1,4-benzoxazin-7-yl.

[094] In another example, R3 isaasubstituted R is substitutedor orunsubstituted unsubstitutedheterocyclic heterocyclicring ringhaving havingthe the

formula:

het het

in which the heterocyclic ring is selected from the group consisting of 5-hydroxy-4 ] H-H-

pyran-4-on-2-yl, 2-hydroxypyrid-4-yl, 2-aminopyrid-4-yl, 2-carboxypyrid-4-yl, or tetrazolo[1,5-

a]pyrid-7-yl.

[095] In one particular embodiment, the tyrosine hydroxylase inhibitor is aquayamycin. In one

example, aquayamycin is a compound of the formula set forth below.

wo 2020/018292 WO PCT/US2019/040581

HO HO OH , OH 0 HO

O OH

0 HO" HO" OH 1

[096] In another particular embodiment, the tyrosine hydroxylase inhibitor is oudenone. In one

example, oudenone is a compound of the formula set forth below.

[097] Other suitable tyrosine hydroxylase inhibitor, known to one of skilled in the art, can also

be used. Example of other tyrosine hydroxylase inhibitor include, for example, but not limited to,

cycloheximide, anisomycin, 3-iodo-L-tyrosine, pyratrione, phenyl carbonyl derivatives having

catechol or triphenolic ring systems, for example, phenethylamine and gallic acid derivatives, 4-

isopropyltropolone, -thiazolyl)benzimidazole, 8-hydroxyquinoline, 2-(4 -thiazolyl)benzimidazole, o-phenantroline, 8-hydroxyquinoline, 5-iodo- o-phenantroline, 5-iodo-

a-a'-dipyridil, 8-hydroxyquinoline, bilirubin, 2,9-dimethyl-1, 10- phenantroline, -'-dipyridil, dibenzo dibenzo

[f,h]quinoxaline, 2,4,6-tripyridil-s-triazine, ethyl 3-amino4H-pyrrolo-isoxazole-5(6H)-

carboxylate, a-nitroso-B-naphthol, sodium diethyldithiocarbamate, -nitroso-ß-naphthol, sodium diethyldithiocarbamate, ethylenediamineteraacetic ethylenediamineteraacetic acid acid

(See R Hochster, Metabolic Inhibitors V4: A Comprehensive Treatise 52 Elsevier (2012)).

[098] In a particular embodiment, the tyrosine hydroxylase inhibitor is L1:79, which refers to a- -

methyl-DL-tyrosine in methyl-DL-tyrosine in aa racemic racemic form form or or D,L D,L -methyl-para-tyrosine a-methyl-para-tyrosine(abbreviated (abbreviatedasasDL-AMPT). DL-AMPT).

WO wo 2020/018292 PCT/US2019/040581

[099] a-methyl-DL-tyrosine inhibits the -methyl-DL-tyrosine inhibits the activity activity of of tyrosine tyrosine hydroxylase hydroxylase (TH), (TH), which which catalyzes catalyzes

the first transformation in catecholamine biosynthesis, i.e., the conversion of tyrosine to

dihydroxyphenylalanine (DOPA), which is the rate limiting step in catecholamine synthesis. a- -

methyl-DL-tyrosine is a tyrosine analog that competes competitively for TH and is excreted mostly

unchanged in the urine.

[100] In preliminary testing, it was observed that Demser (i.e., a-methyl-L-tyrosine) hadaa -methyl-L-tyrosine) had

beneficial effect on the symptoms of leaky gut, but that this effect was short lived. The racemic

mixture (i.e., a-methyl-DL-tyrosine) wasobserved -methyl-DL-tyrosine) was observedto tohave haveaagreater greaterphysiologic physiologiceffect effectand andhalf- half-

life. This may be due to the failure of the D amino acid isomer to be transported by the different

L amino acid transport systems that exist within the body, or by completion for renal excretion

that allows the L isomer to persist for a longer duration.

[101] In another aspect, the invention provides a pharmaceutical composition comprising a

therapeutically effective amount or dose of a tyrosine hydroxylase inhibitor (e.g., a-methyl-DL- -methyl-DL-

tyrosine) and a pharmaceutically acceptable carrier.

[102] Effective doses of the compositions of the present invention, for treatment of autism as

described herein vary depending upon many different factors, including means of administration,

target site, physiological state of the patient, whether the patient is an adult or a child, other

medications administered, and whether treatment is prophylactic or therapeutic. Usually, the

patient is a human but non-human mammals including transgenic mammals can also be treated.

Treatment dosages may be titrated using routine methods known to those of skill in the art to

optimize safety and efficacy.

[103] The pharmaceutical compositions of the invention may include a "therapeutically effective

amount." A "therapeutically effective amount" refers to an amount effective, at dosages and for

periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective

amount of a molecule may vary according to factors such as the disease state, age, sex, and weight

of the individual, and the ability of the molecule to elicit a desired response in the individual. A

therapeutically effective amount is also one in which any toxic or detrimental effects of the

molecule are outweighed by the therapeutically beneficial effects.

[104] As used herein, the terms "treat" and "treatment" refer to therapeutic treatment, including

prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an

WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

undesired physiological change associated with a disease or condition. Beneficial or desired

clinical results include, but are not limited to, alleviation of symptoms, diminishment of the extent

of a disease or condition, stabilization of a disease or condition (i.e., where the disease or condition

does not worsen), delay or slowing of the progression of a disease or condition, amelioration or

palliation of the disease or condition, and remission (whether partial or total) of the disease or

condition, whether detectable or undetectable. Those in need of treatment include those already

with the disease or condition as well as those prone to having the disease or condition or those in

which the disease or condition is to be prevented.

[105] Dosage regimens may be adjusted to provide the optimum desired response (e.g., a

therapeutic or prophylactic response).

[106] In one example, a single bolus may be administered. In another example, several divided

doses may be administered over time. In yet another example, a dose may be proportionally

reduced or increased as indicated by the exigencies of the therapeutic situation. Dosage unit form,

as used herein, refers to physically discrete units suited as unitary dosages for treating mammalian

subjects. Each unit may contain a predetermined quantity of active compound calculated to

produce a desired therapeutic effect. In some embodiments, the dosage unit forms of the invention

are dictated by and directly dependent on the unique characteristics of the active compound and

the particular therapeutic or prophylactic effect to be achieved.

[107] The composition of the invention may be administered only once, or it may be administered

multiple times. For multiple dosages, the composition may be, for example, administered three

times a day, twice a day, once a day, once every two days, twice a week, weekly, once every two

weeks, or monthly.

[108] It is to be noted that dosage values may vary with the type and severity of the condition to

be alleviated. It is to be further understood that for any particular subject, specific dosage regimens

should be adjusted over time according to the individual need and the professional judgment of

the person administering or supervising the administration of the compositions, and that dosage

ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the

claimed composition.

[109] "Administration" to a subject is not limited to any particular delivery system and may

include, without limitation, oral (for example, in capsules, suspensions or tablets), parenteral

WO wo 2020/018292 PCT/US2019/040581

(including subcutaneous, intravenous, intramedullary, intraarticular, intramuscular, or

intraperitoneal injection), topical, or transdermal. Administration to a host may occur in a single

dose or in repeat administrations, and in any of a variety of physiologically acceptable salt forms,

and/or with an acceptable pharmaceutical carrier and/or additive as part of a pharmaceutical

composition (described earlier). Once again, physiologically acceptable salt forms and standard

pharmaceutical formulation techniques are well known to persons skilled in the art (see, for

example, Remington's Pharmaceutical Sciences, Mack Publishing Co.).

[110] In one aspect, the dosage of tyrosine hydroxylase inhibitor may range from about 1 mg to

about 4g. In a particular embodiment, the dosage of tyrosine hydroxylase inhibitor may range

from about 10 mg to about 1500 mg. In some suitable embodiments of the invention, the

composition comprises a tyrosine hydroxylase inhibitor (i.e., a-methyl-DL-tyrosine) in an -methyl-DL-tyrosine) in an amount amount

ranging from about 50 mg (w/w) to about 1500 mg (w/w); from about 50 mg (w/w) to about 500

mg (w/w); from about 75 mg (w/w) to about 350 mg (w/w); from about 90 mg (w/w) to about 350

mg (w/w); from about 100 mg (w/w) to about 300 mg (w/w); or from about 100 mg (w/w) to about

200 mg (w/w). In one embodiment, the composition comprises a tyrosine hydroxylase inhibitor

(i.e., a-methyl-DL-tyrosine) in an -methyl-DL-tyrosine) in an amount amount of of about about 90, 90, 100, 100, 125, 125, 150, 150, 175, 175, 200, 200, 250, 250, 300, 300, 350, 350,

400, 500, 750, 1000, or 1500 mg (w/w/). As used herein, a "composition" refers to any

composition that contains a pharmaceutically effective amount of one or more active ingredients

(e.g., a tyrosine hydroxylase inhibitor, another acne treating agent, or a combination thereof).

[111] In some embodiments, a plurality of compositions having different dosages are

administered concurrently or sequentially. For instance, in one embodiment, a first composition

comprising a-methyl-DL-tyrosine in an -methyl-DL-tyrosine in an amount amount of of about about 200 200 mg mg (w/w) (w/w) and and aa second second composition composition

comprising a-methyl-DL-tyrosine in an -methyl-DL-tyrosine in an amount amount of of about about 100 100 mg mg (w/w) (w/w) are are administered administered

concurrently. In another embodiment, a first composition comprising a-methyl-DL-tyrosine in an -methyl-DL-tyrosine in an

amount of about 200 mg (w/w) and a second composition comprising a-methyl-DL-tyrosine inan -methyl-DL-tyrosine in an

amount of about 100 mg (w/w) are administered sequentially.

[112] In one aspect, the composition is administered for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,

11, or 12 weeks. In another aspect, the composition is administered for a duration of 1, 2, 3, 4, 5,

6, 7, 8, 9, 10, 11, or 12 week dosing period. In yet another aspect, the composition is administered

for a duration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month dosing period.

[113] In another aspect, the invention provides a method for obtaining a plasma concentration of

a a therapeutic therapeutictyrosine hydroxylase tyrosine inhibitor hydroxylase drug (e.g., inhibitor drug a-methyl-DL-tyrosine) for a long for (e.g., -methyl-DL-tyrosine) terma for long term for

treating an autism in a subject in need thereof, the method comprising administering to said subject

said therapeutic drug at a concentration ranging from about 50 mg (w/w) to about 500 mg (w/w)

three times a day, wherein said plasma concentration ranges from about 500 ng/ml to 5000 ng/ml,

and wherein said term is at least 1 week. In one embodiment, to obtain a desired plasma

concentration, the drug is administered for at least 1 day, 2 day, 3 day, 4 day, 5 day, 6 day, 1 week,

2 week, 3 week, or 4 week. In a particular embodiment, to obtain a desired plasma concentration,

the drug is administered for a duration ranging from about 1 day to about 4 weeks; from about 2

days to about 4 weeks; or from about 1 week to about 4 weeks. In some embodiments, to obtain

a desired plasma concentration, the drug is administered for a duration of therapeutic regimens in

excess of 6 months.

[114] The administration of composition of the invention may result in a plasma concentration

ranging from about 500 ng/ml to 5000 ng/ml; from about 800 ng/ml to 2500 ng/ml, or from about

1400 ng/ml to 1800 ng/ml.

[115] The pharmaceutical compositions of the invention may be formulated in a variety of ways,

including for example, solid, semi-solid, and liquid dosage forms, such as capsules, tablets, pills,

powders, liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions,

liposomes and suppositories. In some embodiments, the compositions are in the form of injectable

or infusible solutions. The composition is in a form suitable for oral, topical, intravenous,

intraarterial, intramuscular, subcutaneous, parenteral, transmucosal, or transdermal administration.

In a particular embodiment, the composition is in the form of a capsule. In another particular

embodiment, the embodiment, composition the is in composition isthe in form the of a tablet. form of a tablet.

[116] Administration of the pharmaceutical composition can be through various routes, including

orally, nasally, subcutaneously, intravenously, intramuscularly, transdermally, vaginally, rectally

or in any combination thereof. Transdermal administration can be effected using, for example,

oleic acid, 1-methyl-2-pyrrolidone, dodecylnonaoxyethylene.

[117] In one aspect, the invention provides administering to a subject a therapeutically effective

amount of a first tyrosine hydroxylase inhibitor, for example, a-methyl-DL-tyrosine in -methyl-DL-tyrosine in

combination with a therapeutically effective amount of a second tyrosine hydroxylase inhibitor,

WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

for example, a-methyl-L-tyrosine. Inanother -methyl-L-tyrosine. In anotheraspect, aspect,the theinvention inventionprovides providesadministering administeringto toaa

subject a therapeutically effective amount of one or more tyrosine hydroxylase inhibitors, for

example, a-methyl-DL-tyrosine and/or-methyl-L-tyrosine -methyl-DL-tyrosine and/or a-methyl-L-tyrosine inin combination combination with with a a therapeutically therapeutically

effective amount of another agent useful in the treatment of autism.

[118]

[118] In Insome someofof these aspects, these y-aminobutyric aspects, acid acid -aminobutyric (GABA)(GABA) can be can administered with the with the be administered

tyrosine hydroxylase inhibitor (e.g., a-methyl-DL-tyrosine). The GABA -methyl-DL-tyrosine). The GABA can can be be administered administered

simultaneously with the tyrosine hydroxylase inhibitor. In other aspects, the GABA can be

administered separately from the tyrosine hydroxylase inhibitor, e.g., at another time during the

day. In some aspects the GABA is administered at bedtime. Typically, dosages of the GABA are

from about 5 mg to about 30 mg, for example, 5, 10, 15, 20, 25, or about 30 mg of GABA., with

15 mg of GABA being particularly preferred.

[119] In other of these aspects, a p450 3A4 promoter is administered in addition to the tyrosine

hydroxylase inhibitor (e.g., a-methyl-DL-tyrosine) and the -methyl-DL-tyrosine) and the optional optional GABA. GABA. Preferred Preferred p450 p450 3A4 3A4

promoters include 5,5-diphenylhydantoin, valproic acid, and carbamazepine.

[120] Those subjects on the autism spectrum, including those diagnosed with Asperger's Disorder

(Asperger Syndrome) or Social Communication Disorder, who also have symptoms of ADHD

and/or tics can be treated using methods of the disclosure. In these aspects, the subject can be

administered an effective amount of a tyrosine hydroxylase inhibitor and an effective amount of a

beta adrenergic agonist (also referred to as beta agonists). The tyrosine hydroxylase inhibitor can

be any of the tyrosine hydroxylase inhibitors described herein, with a-methyl-DL-tyrosine being -methyl-DL-tyrosine being

particularly preferred. Beta adrenergic agonists are known in the art and include, for example,

albuterol, levalbuterol, fenoterol, formoterol, isoproterenol, metaproterenol, salmeterol,

terbutaline, clenbuterol, isoetarine, pirbuterol, procaterol, ritodrine, epinephrine, and combinations

thereof. Albuterol is a particularly preferred beta adrenergic agonist.

[121] In certain of these aspects, the tyrosine hydroxylase inhibitor and the beta adrenergic

inhibitor are administered simultaneously. In other aspects, the beta adrenergic inhibitor is

administered separately from the a-methyl-DL-tyrosine, e.g., at -methyl-DL-tyrosine, e.g., at another another time time during during the the day. day.

[122] According to the disclosure, the described methods for treating a disease or disorder can

be used in combination with treatment methods that are also known to be effective in treating the

same disease or disorder. For example, autism behaviors and symptoms can be treated with

25

WO wo 2020/018292 PCT/US2019/040581

compounds that affect autonomic neurotransmission (e.g. amphetamine, methylphenidate, and the

like), psychotopic drugs (e.g., risperidone), neutotransmitter reuptake inhibitors (e.g., fluoxetine),

compounds that stimulate glutaminergic transmission (e.g., LY2140023), and/or compounds that

affect cholinergic neurotransmission (e.g., galantamine). As such, the disclosure is also directed to

methods of treating autism in a patient by administering an effective amount of a tyrosine

hydroxylase inhibitor (e.g., a-methyl-DL-tyrosine) and an -methyl-DL-tyrosine) and an effective effective amount amount of of aa compound compound that that

affects autonomic neurotransmission, a psychotopic drug, a neutotransmitter reuptake inhibitor, a

compound that stimulates glutaminergic transmission, and/or a compound that affects cholinergic

neurotransmission. In one embodiment, the invention provides methods of treating autism in a

patient by administering a therapeutically effective amount of a tyrosine hydroxylase inhibitor

(e.g., a-methyl-DL-tyrosine) and aa therapeutically -methyl-DL-tyrosine) and therapeutically effective effective amount amount of of aa central central nervous nervous system system

(CNS) agent.

[123] The diseases or disorders treated by the composition of the invention include, for example,

autism or its associated disease or disorder.

[124] The autonomic nervous system has been implicated in symptoms that resemble those seen

in autism. Autism spectrum disorder (ASD) has been associated with abnormal findings in

autonomic related structures including the insula and the amygdala. Autonomic related changes

such as increases in basal heart rate and diminished heart rate due to psychosocial challenges are

seen in autism. The autism-autonomic linkage is exemplified by the consequences of respiratory

sinus arrhythmia (RSA) that includes difficulties with socialization, language difficulties, and

delays in cognitive development.

[125] It has been hypothesized a chronically over activated autonomic system is a correlate of

autism based upon the exaggerated levels of anxiety that attend autism, physiologic hyperarousal,

and other correlates. Anxiety, perhaps the greatest co-morbidity associated with autism and which

may drive other features of the disease, has been associated with central nervous system structures

that are linked to autonomic function. Phenotypically autism and anxiety both present with

stereotyped repetitive behaviors and limited interests, avoidance behaviors, and speech problems.

The relationship between anxiety and reported autonomic symptoms of elevated heart rate,

perspiration, and other sequelae of the "fight or flight" reaction reveal a role for the peripheral

nervous system function in autism. However, this may be secondary to central autonomic

WO wo 2020/018292 PCT/US2019/040581

activation. Central functions may manifest as elevated emotional responsiveness and exaggerated

threat perception or diminished inhibition of fear responses, which are associated with the central

structures mentioned above in which autonomic responsiveness and emotional responsiveness

overlap.

[126] There is a considerable body of evidence that associates autism with cholinergic function

in in the the central centralnervous system, nervous specifically system, with various specifically a-subtype-subtype with various nicotinicnicotinic receptors,receptors, notably in notably in

the cerebellum. However, as autonomic function is classically considered to be a balance of

cholinergic and catecholaminergic systems, perceived increases or decreases in cholinergic

function may be manifestations of change in the dynamic balance of these systems with

catecholaminergic tone. It is believed that it may be possible to effect therapeutic change in ASD

through manipulation of either acetylcholine based manipulations or the counterbalancing of

dopamine, norepinephrine, or epinephrine mediated mechanisms by administration of a-methyl- -methyl-

DL-tyrosine.

[127] The present invention also relates to the treatment of the core symptoms of ASD by

administration of a-methyl-DL-tyrosine to subjects -methyl-DL-tyrosine to subjects in in need need thereof. thereof.

[128] While not intending to be bound by any particular mechanism of operation, it is believed

that that the thetyrosine tyrosinehydroxylase inhibitor hydroxylase (e.g.,(e.g., inhibitor a-methyl-DL-tyrosine) administered -methyl-DL-tyrosine) according to administered the according to the

present invention modulates the catecholaminergic pathways implicated in autism, more

specifically such pathways involved in the core symptoms of ASD, including the

catecholaminergic functions in the CNS and in the gastrointestinal tract. Therefore, the known

effects of catecholamines on the endocrine and neuroendocrine systems are regulated by

-methyl-DL-tyrosine), including administration of the tyrosine hydroxylase inhibitor (e.g., a-methyl-DL-tyrosine), including but but

not limited to decreasing the amount of adrenaline secreted into the bloodstream, which may lessen

the intensity of irritability and agitation, and other core symptoms of ASD.

[129] In one aspect, the invention provides a method for treating core symptoms of Autism

Spectrum Disorder (ASD) in a subject in need thereof, the method comprising administering a

therapeutically effective amount of a racemic mixture of a-methyl-DL-tyrosine. -methyl-DL-tyrosine.

[130] In another aspect, the invention provides a method for treating an autism associated clinical

trait in a subject in need thereof, the method comprising administering to said subject a

WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

composition comprising a therapeutically effective amount of a tyrosine hydroxylase inhibitor

(e.g., a-methyl-DL-tyrosine), thereby treating -methyl-DL-tyrosine), thereby treating said said autism autism associated associated clinical clinical trait trait in in said said subject. subject.

[131] Examples of an autism associated clinical trait include, for example, but not limited to, a

lack of social communication, a lack of social interaction, a lack of social motivation, lethargy and

social withdrawal, inappropriate speech, hyperactivity, stereotypic behavior, irritability and

agitation, restrictive behavior, repetitive behavior, ritualistic behavior, sameness behavior,

compulsive behavior, self-injurious behavior or a combination thereof.

[132] In one embodiment, the clinical trait is assessed based upon a change from a baseline in

one or more psychometric tests. Examples of a psychometric test include, for example, but not

limited to, clinical global impression (CGI) rating scale, Vineland adoptive behavior scale

(VABS), autism diagnostic observation schedule (ADOS), social responsiveness scale (SRS),

aberrant behavior checklist - community (ABC-C), repetitive behavior scale (RBS), Conners

parent rating scale (CPRS), or a combination thereof. In a particular embodiment, the clinical trait

meets the requirements of Diagnostic and Statistical Manual of Mental Disorders - V (DSM-V)

criteria.

[133] The terms "subject," "individual," and "patient" are used interchangeably herein, and refer

to an animal, for example a human, to whom treatment, including prophylactic treatment, with the

pharmaceutical composition according to the present invention, is provided. The term "subject" as

used herein refers to human and non-human animals. The terms "non-human animals" and "non-

human mammals" are used interchangeably herein and include all vertebrates, e.g., mammals, such

as non-human primates, (particularly higher primates), sheep, dog, rodent, (e.g. mouse or rat),

guinea pig, goat, pig, cat, rabbits, cows, horses and non-mammals such as reptiles, amphibians,

chickens, and turkeys.

[134] In one embodiment, the subject is a human patient between 3 years of age and 21 years of

age; 5 years of age and 21 years of age; or 6 years of age and 17 years of age. In another

embodiment, the subject is an adult human patient.

[135] All patents and literature references cited in the present specification are hereby

incorporated by reference in their entirety.

WO wo 2020/018292 PCT/US2019/040581

[136] Also provided herein are kits comprising one or more molecules or compositions described

herein. The following examples are provided to supplement the prior disclosure and to provide a

better understanding of the subject matter described herein. These examples should not be

considered to limit the described subject matter. It is understood that the examples and

embodiments described herein are for illustrative purposes only and that various modifications or

changes in light thereof will be apparent to persons skilled in the art and are to be included within,

and can be made without departing from, the true scope of the invention.

EXAMPLES EXAMPLE 1

[137] Two-hundred patients were initially screened. Thirty subjects meeting the study criteria

consented. Nine (9) subjects had high blood glucose levels (hyperglycemia) prior to consenting to

the study.

[138] A high blood glucose level (hyperglycemia) is defined as a fasting plasma blood glucose

level of 126 mg/dl or greater on two separate occasions. The average patient age was sixty- two

years old and the median patient age was sixty years old. Six of the patients were female and three

of the patients were male. Five of the patients were fifty to sixty years old and four of the patients

were over the age of sixty.

[139] The patients in the study were administered a treatment regimen that included a tyrosine

hydroxylase inhibitor (i.e., a-methyl-DL tyrosine),aamelanin -methyl-DL tyrosine), melaninpromoter promoter(i.e., (i.e.,melanotan melanotanII), II),aap450 p450

3A4 promoter (i.e., 5, 5-diphenylhydantoin), and a leucine aminopeptidase inhibitor (i.e., N-

[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine) These

[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryI]-L-leucine). These compounds compounds were were administered administered

on each of five days per week for a period of six weeks, with one or two days off between weekly

cycles. Blood glucose level was monitored for all subjects biweekly. Blood glucose levels were

determined by daily blood glucose tests followed-up with laboratory blood glucose tests every two

weeks.

[140] After approximately two to four weeks, all nine of the subjects had normal blood glucose

levels defined as a fasting plasma blood glucose level of 125 mg/dl or lower on two separate

occasions.

WO wo 2020/018292 PCT/US2019/040581

[141] Overall, the above-noted treatment was well tolerated by the subjects, with no adverse

events related to the treatment, and responses have been documented to the treatment 100%.

EXAMPLE 2

[142] Patients are screened and the extent to which they meet the DSM-V criteria for autism

spectrum disorder is assessed. A subgroup of those satisfying the criteria are administered a

treatment regimen that includes a tyrosine hydroxylase inhibitor (i.e., a-methyl-DL tyrosine) at -methyl-DL tyrosine) at

dose of 50-100 mg three times daily. Another subgroup is administered a treatment regimen that

further includes a p450 3A4 promoter (i.e., 5, 5-diphenylhydantoin) in one daily dose of 30 mg.

Gamma- aminobutyric acid is optionally administered to both subgroups at bedtime at a dose of

15 mg to aid sleeping and to quiet ticks and repetitive behaviors like teeth grinding. Vasopressin

is administered as needed to assist brain governance. Following each administration of the

treatment regimen, changes in the extent to which the subjects satisfy the DSM-5 criteria is again

assessed.

EXAMPLE 3

[143] A Blinded, Randomized, Placebo Controlled Phase 2 Study for the use of AMPT in

the Treatment of Autism

[144] Study Design: Blinded, randomized, 2 arm, 8-week treatment period followed by

additional follow-up visits off treatment over the next 18 weeks for a total of 26 weeks' study

participation. Treatment participation. armsarms Treatment consist of LI of consist -79LI-79 alone alone or placebo. or placebo.

[145] Sample Size:L1-79 N=30, placebo N=10

[146] Study Population: Autistic patients over 12 years of age that meet the entry criteria and

who are high performing in the opinion of the investigator.

[147] Major Inclusion Criteria: Signed informed consent, normal clinical laboratory values,

DSM-5 compliant diagnosis of autism, Qualifying ADOS score, sufficiently high functioning to

complete the protocol, no psychotropic drugs for at least 2 weeks, ABC-C score >12.

[148] Major Exclusion Criteria: Fragile-X syndrome, epilepsy, use of complimentary alternative

medications, any co-morbidities, other psychiatric disorder, out of range lab values.

WO wo 2020/018292 PCT/US2019/040581

[149] Experimental Treatment: LI -79 is a racemic form of the drug Demser ItIt Demser®. will bebe will given given

daily X x 8 weeks, and then followed on weeks 10, 13 and 26.

[150] Non-Experimental Treatment: Placebo.

[151] Dosage and Administration: LI -79 or placebo, as randomized, will be administered orally

at a dose of 90 mg TID.

[152] Evaluation Schedule: Patients will receive 8 weeks of the schedule above with weekly

treatment evaluations for weeks 1-8 then post-treatment follow up visits at weeks 10, 13 & 26.

[153] Safety Measures: Regularly scheduled complete history and physical examination, vital

signs, CBC, differential, platelet counts, urine analysis, serum enzymes including: total protein,

albumin, glucose, BUN, creatinine, direct and total bilirubin, alkaline phosphatase, phosphorous,

calcium, aspartate aminotransferase ("AST"), alanine aminotransferase ("ALT"), sodium,

potassium, chloride, bicarbonate, T4, TSH, and adverse events assessments. An independent DMC

will oversee the conduct of this trial to assure patient safety.

[154] Study Duration: A maximum of 50 weeks (12 weeks' enrollment, 38 weeks' treatment and

follow-up).

[155] Study Endpoints: The primary end point will be the assessment of the attending physician

as reflected in the Clinical Global Impressions (CGI) scale based upon changes from baseline in

various psychometric tests, including the Aberrant Behavior Checklist-Community (ABC-C),

Conners Parent Rating Scale and the Autism Diagnostic Observation Schedule (ADOS), as well

as from their personal observations in the clinic, and from videographic information taken at

regularly scheduled clinic visits (per this protocol) and provided by caregivers over the course of

this study.

EXAMPLE 4

[156] a-methyl-DL tyrosine(AMPT) -methyl-DL tyrosine (AMPT)was wasadministered administeredto to33patients. patients.This Thisgroup groupof ofpatients patientswas was

qualified under the DSM-5 definition of autism and treated. See Table 1.

[157] Table 1

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Patient Age Sex Dosage Adverse number Events Events AMPT AMPT 01-001 3 Male Male 90 TID None 01-002 15 Male 90 TID None 01-003 11 Male Male 90 TID None

The Aberrant Behavior Checklist - Community (ABC-C), Autism Diagnostic Observation

Schedule (ADOS), Conners Parent Rating Scale (CPRS), and General Clinical

Impressions scale were used to assess the disease. Videos were taken at each visit.

[158]

[158] Data Data for for two two of of these these patients patients over over the the first first 33 weeks weeks of of this this observation observation are are presented presented in in the the

tables below. Conners Patent Rating Scale Data is depicted in Figure 1. These clinical

improvements appear to begin quickly and are durable. Continued improvement over weeks and

months has been observed. No adverse events have been reported other than mild tiredness on the

first day of dosing in two patients.

Table 2

PT LR Date of Assessment initial Day Test Dimension Day 7 Day 14 Day 21 21 Day 28

ABC-C 114 71 55 53 53 ABC-C TOTAL irritability 21 16 16 16 16 lethargy 31 12 8 7 7 7 stereotypy 13 5 0 0 0 hyperactivity 34 31 24 23 23 speech 5 " 7 7 7 7

DSM-V 42 27 21 18 18 TOTAL social 9 5 3 2 2 communication 10 4 3 2 2 relationships 11 11 4 3 2 2 behavior 12 12 12 12 12

WO wo 2020/018292 PCT/US2019/040581

Table 3

PT RS Date of Assessment Test initial Day 21 Dimension Day 7 Day 14 Day 28 53 28 20 20 20 ABC TOTAL irritability I1 II II 3 3 lethargy 11 I- I 1 6 3

stereotypy 16 11 9 9 9 hyperactivity 20 8 6 6 6 speech 8 3 3 3 3

21 15 8 6.5 5.5 DSM V TOTAL social 5 5 3 1.5 I- ]1

communication 5 4 1.5 1.5 1.5

relationships I 0 0 0 0 or in is behavior 10 8 5 4 3

Table 4

PT WC Date of Assessment PTWC Test Dimension initial Day 7 Day 14 Day 21 Day 28 Day28 70 30 18 18 17 17 17 ABC-C TOTAL irritability 16 in 16 4 2 2 2 lethargy 21 9 3 2 2 stereotypy 12 7 6 6 6 6 hyperactivity 20 20 10 7 7 7 II speech 0 0 0 0 0

26 17 14 12 11 DSM-V TOTAL social 6 6 4 2 2 communication 3 3 0 0 0 0 relationships 6 2 2 2 2 2 behavior 11 11 8 X 8 7 9

EXAMPLE 5 Evaluation of L1-79 Administration in Patients with Autism

[159] L1-79 was used anecdotally in two patients followed by a more structured evaluation in 8

additional patients with autism. All patients were administered a starting dose of L1-79 of 90 mg

TID for a minimum of 3 months. During the evaluation doses as high as 400 mg TID were used.

A summary of data available on these 10 patients was provided in the Summary of Clinical

Efficacy, Use of L1-79 to treat autism, submitted to IND 128673, sequence number 0005 0005.While While

the sample size was small and involved open-label administration of L1-79, the results were

WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

encouraging with consistent improvements in the ABC-C domains and an average reduction in the

ADOS-2 of 30%, and one child manifesting an ADOS-2 decrease of 60% which resulted in the

loss of his autism diagnosis. In addition, longer treatment resulted in a greater magnitude of

therapeutic benefits on the core symptoms of autism. Moreover, when the study drug, L1-79, was

discontinued not all of the benefits regressed. These data suggest that L1-79 has the potential to

improve the core symptoms of autism.

EXAMPLE 6 A Randomized, Double-Blind Placebo-Controlled 4-week Study in Male Patients Diagnosed with Autism

[160] The purpose of this clinical study (referred to herein as "Study HT 02-121"; i.e., Example

6) was to determine whether L1-79 was a well-tolerated and clinically useful agent for the

treatment of ASD, and to assess the PK and pharmacodynamics (PD) of four weeks of TID dosing

with L1-79. While the preliminary study of Example 5 involved open-label treatment with L1-79

for at least three months, the present study was only 4-weeks based upon limitations in the

available toxicology data. Based on the shorter duration, the response to L1-79 was expected to

be less in Study HT 02-121 compared to the preliminary study of longer duration.

[161] This clinical study was a randomized double-blind, placebo-controlled two-cohort, 4-week

dose-escalation study that incorporated two open-label treatment groups to assess the safety and

efficacy of L1-79 100 mg and 200 mg TID in male patients between the ages of 12 - 21 years of

age diagnosed with autism. The first cohort of 20 patients was comprised of three groups of

patients: The first group of five patients received open-label L1-79 100 mg TID and underwent

PK and EKG assessments. The remaining two groups in this cohort consisted of 15 patients

randomized on a 2:1 basis to receive L1-79 100 mg TID or placebo. The PK and safety data from

the open-label L1-79 100 mg TID group was submitted to FDA for review and acceptance before

the second cohort was enrolled. The second cohort was procedurally identical to the first. The

second cohort of 20 patients was comprised of the same three groups of patients but a dose of 200

mg TID was used instead of 100 mg TID. The key inclusion criteria were as follows: (1) males

between 13 and 21 years of age, (2) DSM-5 compliant diagnosis of autism spectrum disorder,

confirmed by the Autistic Diagnosis Interview Review (ADIR), and by an ADOS-2 score consistent with a diagnosis of autism, (3) must have been stable on no more than one concomitant medication and no planned changes in psychosocial interventions during the study.

[162] The key exclusion criteria were as follows: (1) sexually active males, (2) a history of

Fragile-X syndrome, Rett syndrome or any other co-morbidity including but not limited to cancer,

genetic diseases, or any disease or syndrome that required drug therapy, (3) DSM-5 diagnosis of

schizophrenia, schizoaffective disorder, alcohol use disorder or attention deficit hyperactivity

disorder (ADHD), (4) had any active medical problem(s), including epilepsy and asthma, (5)

uncontrolled intercurrent illness including, but not limited to, ongoing or active infection,

symptomatic cardio-vascular disease, hepatic disease, renal disease, skeleto-muscular disease,

human immunodeficiency virus (HIV), hemorrhagic cerebrovascular accident (HCVA), hepatitis

B virus (HBV), or psychiatric illness/social situations that would limit compliance with study

requirements, (6) any disease that required chronic treatment, (7) any disease that required

treatment with an immunosuppressive drug, and/or (8) current or lifetime diagnosis of severe

psychiatric disorder.

Open-label Patients

[163] The first five patients in each cohort were assigned to receive active medication in an open-

label fashion. The patients were treated identically to the randomized patients with the following

exceptions: (1) blood samples were drawn at baseline and 1 hour after dosing, and at Week 1, 2, 3

and 4 treatment visits 1 hour after dosing to determine the PK of L1-79, (2) EKGs were assessed

at baseline and within 3-days prior to the Week 1, 2, 3 and 4 treatment visits as well as the 1-week

and 4-week post-dosing follow-up visits, (3) vital signs, physical exams, and ASD assessments

were performed at 1-week postdosing visit, (4) blood and urine samples were drawn at baseline

and at Week 1, 2, 3 and 4 treatment visits 1 hour after dosing as well as the 1-week and 4-week

post-dosing follow-up visits for safety analyses.

Randomized Patients

[164] The randomized patients were treated identically to the open-label patients with the

following three exceptions: (1) no blood samples for PK were drawn, (2) no ECG was performed,

and (3) these patients did not have a 1-week follow-up visit, only a 4-week follow-up visit. A time

and events schedule for the study is provided below in Table 5.

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Safety Endpoints

[165] Safety was the primary endpoint of the study. The following safety endpoints were

assessed: ECGs, physical exams, laboratory evaluations (hematology, chemistry and urinalysis),

vital signs, including orthostatic blood pressure, adverse events (AEs) and concomitant

medications.

Efficacy Endpoints

[166] The primary efficacy endpoint was the determination of the clinical improvement by the

investigator as documented by the Clinical Global Impression (CGI) rating scales. Additional

efficacy endpoints included: (1) changes from baseline in the Socialization and Communication

Domains of the Vineland Adaptive Behavior Scales, Second Edition (VABS II) parent/caregiver

rating form, (2) changes from baseline in the ADOS-2 Total Score and subscores, (3) changes in

the Social Responsiveness Scale, Second Edition (SRS-2) Total Score and subscales, (4) changes

in the ABC-C domains, (5) changes in the Repetitive Behavior Scale - Revised (RBS-R) Total

Score and subscales. Given the exploratory design of this study only descriptive statistics were

planned.

Results

Study Population

[167] Patient enrollment and disposition are summarized in Figure 2. A total of 42 patients were

randomized and received at least one dose of study drug. One patient was randomized to open-

label L1-79 200 mg TID but the parent requested voluntary withdrawal from the study at Week

0/Baseline after receiving a single dose of study medication in clinic. Thirty-nine patients

completed the study. One patient treated with double-blind L1-79 200 mg TID withdrew from the

study due to an AE (see Table 8) and one patient treated with placebo voluntarily withdrew from

the study. Demographic characteristics are summarized in Table 6.

Characteristics Demographic 6 Table Characteristics Demographic 6 Table L1-79 L1-79:

L1-79100 L1-79 200

100mg 200 mg Placebo

mgTID mg TID TID

TID Placebo

Double

Open Open

Double-blind Doubleblind

Double-blind blind

Open label Open label label

label N=5'

N=5 N=5' N=102

N=11

N=5 N=11

N=10²

Age WO 2020/018292

Age (years) (years)

Mean 16.4(2.7)

Mean (SD) 16.2(2.9) 15.8(2.7)

16.4(2.7) 15.8(2.7)

16.4(1.1)

(SO) (2.2)

16.2(2)

Range Range(min, (min,max) max) 12,20

13,20 15,18

Sex, (%) 5(100) 5(100)

10(100) 10(100)

11(100)

Race, (%)

(%) Caucasion 4 (80) (100) 7(70)

9(82)

10(100)

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1 (20) 1(10)

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Safety

Concomitant Medications and Physical Exams

[168] The majority of patients were not on CNS medications during the study. The following

CNS medications were used during the study: clonidine (n=2), Strattera@(n=1), Strattera®(n=1), Depakote Depakote®(n=3), (n=3),

lorazepam (n=1), Prozac Prozac®(n=1), (n=1),and andAbilify (n=1). Abilify® There (n=1). were There nono were clinical significant clinical physical significant physical

exam findings reported during the study.

Adverse Events

[169] The incidence of AEs by primary system organ class and preferred term is presented in

Table 7. and a listing of the AEs reported during the study is presented in Table 8. All AEs were

mild to moderate in intensity and self-limited.

1(10.0)(1)(16.7) 1(10.0)(1)(16.7) 1(10.0)(1)(16.7) 1(10.0)(2)(33.3) 1(10.0](1](16.7) 1(10.0)(2)(33.3) 3(30.0)(6)(100) Allacadio Placebo

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WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

[170] Of the patients who experienced an AE (34.1%, 14 of 41 patients), the majority experienced

AEs of mild (24.4%, 10 of 41 patients) to moderate (14.6%, 6 of 41 patients) intensity. Only three

AEs were reported by more than one patient. Intentional self-injury was reported as a treatment

emergent AE by two patients in the open-label 100 mg group and one patient in the double-blind

placebo group. Irritability was reported as a treatment emergent AE by two patients in the open-

label 100 mg group. For one of the patients reporting intentional self-injury and irritability (02-

002), these symptoms emerged within a few days of withdrawal from active treatment (i.e., after

completion of Week 4). For the other patient (02-001), symptoms began while on active drug

treatment. Urine cystine crystal present was reported as a non-treatment emergent AE at the 4-

week follow-up visit by two patients in the double-blind 200 mg group.

[171] There were no serious adverse events (SAEs) or deaths reporting during the study. A total

of 2 patients were withdrawn from the study due to adverse events: 1 patient in the double-blind

200 mg group experienced a treatment emergent AE of grand mal seizure and 1 patient in the

double-blind placebo group experienced treatment emergent AEs of intentional self-injury and

agitation.

Vital Signs

[172] Orthostatic vital signs were taken at each visit as described in Study HT 02-121. There

were no changes observed that met criteria for orthostatic hypotension (i.e., drop in systolic blood

pressure of 20 mm Hg, drop in diastolic blood pressure of 10mm hg, or increases in heart rate of

30 beats per minute). A small number of subjects demonstrated asymptomatic drop in systolic

and/or diastolic blood pressures, along with increase in heart rate at Week 1 or Week 2 of

treatment, but these resolved by Week 4 and were mild in nature (e.g., a drop of 10 mm Hg in

systolic blood pressure, 5 mm Hg in diastolic pressure and increase of 15 - 24 beats per minute in

heart rate). Figure 3 provides an example of the largest change from baseline that was noted, and

demonstrates that these changes were transient, falling far short of orthostatic criteria.

ECGS and Laboratory Evaluations

[173] No clinically significant changes from baseline EKGs were noted in any of the patients.

Urinalysis revealed no findings related to clinical symptoms. Of note was the observation that half

(50%) of the patients on L1-79 developed crystalluria compared to 30% of placebo treated patients.

Of those that developed crystalluria during the study, approximately half (50%) had concentrated

WO wo 2020/018292 PCT/US2019/040581 PCT/US2019/040581

urine (specific gravity > 1.025). Most of the crystalluria consisted of calcium oxalate crystals.

There were no symptoms associated with the crystalluria.

[174] One patient developed asymptomatic transient mild elevation of amylase at Week 4 (146

U/L, upper limit of normal [ULN] is 125 U/L), which resolved by the 4-week follow up visit.

[175] There were no significant elevations in AST or ALT or any other chemistry parameters

noted in the safety population.

[176] Assessment of hematology parameters revealed no significant deviations in the safety

population.

Pharmacokinetics

[177] Pharmacokinetics were assessed in the open-label 100 mg and 200 mg TID patients. On

Day 1, L-79 concentrations were assessed one-hour post dose. Subsequently, a random L1-79

plasma concentration was assessed at Week 1, Week 2, Week 3 and Week 4. At 1-hour post-dose

on Day 1 plasma concentrations with 100 mg and 200 mg TID ranged from 0 (<2.5) to 736 ng/mL

and 23 to 1680 ng/mL, respectively. A summary of individual patient and combined overall mean

L1-79 random plasma concentrations are shown in Figure 4. As shown in Figure 5 and Figure

6, overall, random plasma L1-79 concentrations were relatively stable over Week 1 to Week 4.

Efficacy

[178] At the time this study was initiated, there was a lack of long-term, juvenile and reproductive

toxicology data. Applicants embarked on a short study of 28 days in duration, with a limitation in

the number, gender, and age of the patients allowed into the study. As a result, this study was not

designed or powered to demonstrate statistical significance on any of the efficacy endpoints. Thus,

as expected none of the outcomes measures achieved statistical significance. What was

anticipated, and what was observed, were positive trends in a variety of instruments consistent

with an improvement in the core symptoms of autism. In fact, there were multiple efficacy

measures demonstrating similar indicators of improvement in the treatment of target core social

domains affected by ASD. In addition, efficacy data are only displayed for patients that received

blinded treatment.

WO wo 2020/018292 PCT/US2019/040581

Outcome Measures

[179] There were some differences in how questionnaires were administered during Cohort 1 and

Cohort 2 of the study. In the preliminary study, all forms were completed at the study site in the

presence of the investigator. In Cohort 1 of the present study, the VABS-II, SRS-2, ABC-C and

RBS-R were filled out at home by the patient/patient's families prior to treatment visits and

returned at treatment visits in order to expedite the execution of the study. The assessment

procedures for VABS II, SRS-2, ABC-C and RBS-R during Cohort 1 were not done at the

treatment visits and as a result, families had difficulties filling out the forms properly. In order

improve the quality and completeness of the questionnaires during Cohort 2, all questionnaires

(with the exception of the ADOS-2 [administered by external certified ADOS-2 test administrator

for both cohorts] and CGI [completed by investigator in both cohorts]) were completed by the

principal investigator with the assistance of the patient/patient's families at the pre-specified

treatment visits. As the Cohort 2 data was more robust, the discussion of results focus on the

comparison of L1-79 200 mg to placebo, with the exception of the ADOS-2 and CGI, where both

L1-79 100 mg and 200 mg are compared to placebo.

[180] After the conclusion of the study, it was discovered that the basal anchors for the VABS-

II were not always appropriately established. In addition, only two subdomains (communication

and socialization) were completed for the patients, making it impossible to obtain domain or total

scores. However, the results for the VABS II communication and socialization domains are

presented below.

[181] A tabular summary of the efficacy measures is presented below (Table 9). For those

measures demonstrating positive trends in favor of L1-79 three sets of figures are presented for

each efficacy measure. The figures consist of the following: a) a line graph showing the

comparative change between the 3 treatment groups from screening/baseline to 4 weeks (followed

by a 4-week post treatment timepoint in some cases), b) an individual patient response plot, and c)

a responder analysis plot. For those efficacy measures not showing a positive trend in favor of L1-

79, only a line graph showing the comparative change between the 3 treatment groups from

screening/baseline to week 4 (followed by a 4-week post treatment timepoint in some cases) is

presented.

WO wo 2020/018292 PCT/US2019/040581

Responder Definition:

[182] In addition to analyzing the data by observing the magnitude of change in each efficacy

endpoint for patients exposed to active drug compared to placebo, it is important to define a

'responder' population. In general, responder analyses are intended to focus on the number of

patients demonstrating any benefit, rather than the overall change in the studied population.

Typically, responders are defined as patients whose target symptoms demonstrate a pre-specified

improvement in the efficacy endpoint. This definition is consistent with accepted standards for

judging mid and long term outcomes for experimental treatments studied in nearly every disorder,

including autism. However, it does not take into account the need for a refined definition in studies

of short duration for disorders, like autism, that have oscillations in symptom severity. In ASD,

children often go through periods of mild to moderate improvement or worsening in their

behavioral, social and emotional symptom regulation related to a variety of identifiable and as yet

unidentified factors (Boso, 2010; Jyonouchi, 2011). Thus, in a study of short duration, a 'baseline'

measure may in fact be at the apex, midpoint or nadir of one of these oscillations. If they are at an

apex or mid-point when the target symptoms under study are measured at baseline, then a short

duration study may only have the opportunity to demonstrate initial efficacy by preventing the

target symptom from returning back to its oscillatory nadir. To account for this probable

occurrence among some study patients, the responder definition includes patients demonstrating

either short term stability or improvement in the target symptoms. Thus, for each efficacy endpoint

demonstrating a positive trend, responder analyses were conducted as defined herein.

[183] A tabular summary of the change from screening/baseline for efficacy measures is

provided in Table 9.

Measures Efficacy for Placebo and L1-79 Between Screening/Baseline from Changes Mean of Comparisons Summary 9 Table Measures Efficacy for Placebo and L1-79 Between Screening/Baseline from Changes Mean of Comparisons Summary 9 Table LOCF. or 4 Week at LOCF. or 4 Week at L1-79 Double-blind L1-79 Double-blind L1-79 Double-blind L1-79 Double-blind 200

100 200mg

100 mg Placebo Placebo

mgTID

mg TID TID

TID N=9

N=9 N=11 N=9

N=9

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9 3

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effect treatment for P-value effect treatment for P-value 0.16

1.00 0.16 N/A N/A

1.00

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in :- -1.1

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-0.3 FIL 0.5)

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0.27 0.40

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7

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(SD) Baseline from Change Mean 0.8 (1.5)

4.2(12.1) (1.5)

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46 6.8 (42. 17.8)

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0.18

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7

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effect treatment for P-value effect treatment for P-value 0.70 0.37 N/A N/A

0.37

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9 8

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-3,-1,0 -4,-2,-1

& 0,0,4 3-10 -1

CI) (95% placebo versus effect Treatment 7.1) (-2.0, 2.5 CI) (95% placebo versus effect Treatment 2.9) (-5.1, -1.1 N/A N/A

25 (20, 7.1) -8.1(-5.1.2.9)

effect treatment for P-value effect treatment for P-value 0.26 0.57

0.26 N/A

0.57 N/A

(N) Score Total RBS-R (N) Score Total RBS-R 11

to 9 8

(SD) Baseline from Change Mean (SD) Baseline from Change Mean -32 -76.

-3.2 -16.1

(7.8) (24.1) -11.0(11.0)

(7.8) -11.0 (11.0)

(24.1)

max) (min, Range max) (min, Range -14, 12 -78. -24.

-14.12 -78, 1 -24, 6

75") median, (25th) Quartiles 75*) median, (25) Quartiles -21.12.2

-6,4,-1 -28, 2 0

28.20 -12, 21, &

S.A.

CI) (95% placebo versus effect Treatment 5) 17.5) (-2.0, 7.8 12.5) (-22.7, -5.1 CI) (95% placebo versus effect Treatment N/A

422.7. 12.5)

.8(-2.0, 17.5)

effect treatment for P-value effect treatment for P-value 0.11 0.55

0.11 N/A N/A

0.55

(N) Behavior Restricted RBS-R (N) Behavior Restricted RBS-R 11 2

S 7

$ (SD) Baseline from Change Mean (SD) Baseline from Change Mean 1.3/2.6) -1.6(1.6)

1.3 (2.6) -1.6 (1.6)

-2.5 (2.8) 25(2.8)

max) (mirs, Range max) (min, Range .1. S -8. 0

-S. 4.0

1,8 4. ©

75m) median, (25th) Quartiles 75*) median, (25, Quartiles o. 1. 1 -3,-2,0

-5,-1,0

0,1,1 5.00 3.20

CI) (95% placebo vensus effect Treatment 2.9

CI) (95% placebo versus effect Treatment -1.0

2.9(0.5 -1.0634 N/A

(0.5, N/A

5.41 (341.5)

54) 1.5)

effect treatment for R-value effect treatment for P-value 0.41 N/A

0.41

0.023 N/A

0.023

(N) Behavior Ritualistic RBS-R (N) Behavior Ritualistic RBS-R 11 7

30 11

9

(SD) Baseline from Change Mean (SD) Baseline from Change Mean 0.1 -3.7

0.1(2.8) -3.1((4.1) -3.7(3.5)

(2.8) (3.5)

-3.1 (4.1)

max) (min, Range max) (min, Range -13,0 0

-13,0 -9.0 -9,0 0

A4, *4

75mg median, (25mg) Quartiles 75m) median, (25, Quartiles -7.

-$ -7,-3,0 -3. o

-5,-1,0 0

5.00 PCT/US2019/040581

a 2 -1,0,2

CI) (95%) placebo versus effect Treatment CI) (95% placebo versus effect Treatment 7.2) (0.5, 3.8 4.6) (-3.4, 0.6 N/A

0.6 634 4.6)

3.8 (O.S. 7.2)

effect treatment for E-value effect treatment for P-value 0.74 N/A N/A

0.028 N/A

0.028 0.24

Measures Efficacy for Placebo and L1-79 Between Screening/Baseline from Changes Mean of Comparisons Summary 9 Table Measures Efficacy for Placebo and L1-79 Between Screening/Baseline from Changes Mean of Comparisons Summary 9 Table LOCF. or 4 Week at LOCF. or 4 Week at L1-79 Double-blind L1-79 Double-blind L1-79 Double-blind L1-79 Double-blind wo 2020/018292

100 mg TID 200 mg TID Placebo

200 mg TID

100 mg TID N=9

N=9 N=9

N=11 (N) Behavior Sameness RBS-R (N) Behavior Sameness RBS-R 7

11

9 11 (SD) Baseline from Change Mean (SD) Baseline from Change Mean -0.7 -0.7 (2.0) (2.0) -2.9 (2.2)

-4.2 (7.1) -2.9(2.2)

4.2 (7.1)

Range Range (min, -22, 1

4.2 -6, 0

22 1 -6.0

(min, max) 4, 2

max) 75th) median, (25th, Quartiles 75*) median, (25*, Quartiles -5,-2,-1 -1

-9,0

-1.-1.0 -1,-1,0 5.2

0 -9,0,0 CI) (95% placebo versus effect Treatment CI) (95% placebo versus effect Treatment 2.2(-0.1.4.4) -1.3(-6.3,3.6) -1.3 (-6.3, 3.6)

2.2 (-0.1, 1, 4.4) effect treatment for P-value effect treatment for P-value 0.57

0.056 N/A N/A

0.056 (N) Behavior Compulsive RBS-R (N) Behavior Compulsive RBS-R 11

9 7

(SD) Baseline from Change Mean (SD) Baseline from Change Mean -1.2 (3.6) -1.6 (1.7)

-2.7(4.9)

-1.2(3.6) -1.6(1.7)

-27 (4.9)

Range Range (min, -16,1 0 -16,0

8.3 4.0

(min, max) -4,0

max)

49 $ 75th) median, (25th Quartiles 75*) median, (25th, Quartiles 2.0 3.0.0 -3,-1,0 3-1,0

1 -3,0,0

-2,0,1 CI) (95% placebo versus effect Treatment CI) (95% placebo versus effect Treatment 0.3 -1.2(-4.6,2.3)

0.3 (-2.8,3.5) (-2.8, 3.5) -1.2 (-4.6, 2.3)

effect treatment for P-value effect treatment for P-value 0.49 N/A N/A

0.49

0.82 (N) Behavior Stereotypic RBS-R (N) Behavior Stereotypic RBS-R 11 7

9 11

(SD) Baseline from Change Mean (SD) Baseline from Change Mean -0.6 -0.6 (1.7) -0.4 (4.2)

(1.7) -0.4 (4.2)

-1.8 (2.8) -1.8(2.8)

Range Range (min, -7,0 -7,6

(min, max) max) -7,0

-5, who in 75th) median, (25th, Quartiles 75*) median, (25th, Quartiles 0.0.0 3,0,3

0,0,0 -5,0,0 -5,0,0 -3,0,3

CI) (95% placebo versus effect Treatment CI) (95% placebo versus effect Treatment -0.1 1.4(-4.8,2.1)

-0.1 (-4.1,3.8) -1.4 (-4.8, 2.1)

(-4.1, 3.8)

effect treatment for P-value effect treatment for P-value 0.41

0.94 N/A N/A

0.94 0.41

(N) Behavior Injurious Self RBS-R (N) Behavior Injurious Self RBS-R your

11

9 7

(SD) Baseline from Change Mean (SD) Baseline from Change Mean -1.7 -1.7 (3.9) (3.9)

-2.2 (5.3)

-2.2(5.3) -2.1(3.8) -21 (3.8)

Range -16,

Range (min, -16, 11 -12, 1 -8, 2

(min, max) -8,2

-12.

max) 75th) median, (25th, Quartiles 75*) median, (25th, Quartiles -7,0,0

200 -1,0,0

-2,0,0 -1,0,0

CI) (95% placebo versus effect Treatment CI) (95% placebo versus effect Treatment -0.1 44-(3.5,4.4)

-0.1 (-5.2,5.0) 0.4 (-3.5, 4.4)

(-5.2, 5.0)

effect treatment for P-value effect treatment for P-value 0.97 0.97 0.83 N/A N/A

0.83 PCT/US2019/040581

forward carried observation LOCF=last forward carried observation LOCF=last

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Clinical Global Impression (CGI) Rating Scales

[184] The CGI - Overall Severity (CGI-S) and CGI - Overall Improvement (CGI-I) for L1-79

compared to placebo over time are displayed graphically in Figure 7 and Figure 8. The mean

CGI-S change from baseline at Week 4 or last observation carried forward (LOCF) is displayed in

Figure 9, and demonstrates a mean 0.5 point improvement on the overall CGIS for patients in the

200 mg group, compared to those in the placebo and 100 mg groups.

[185] A summary of the change in CGI-S from Baseline to Week 4 is displayed by patient in

Figure 10. In addition, responder analyses (defined as improvement) are presented for CGIS at

Week 4 or LOCF in Figure 11.

[186] Responder definitions for CGI-S were defined as improvement only (instead of

improvement or no change) since, unlike the other measures which were performed weekly, the

CGI-S requires the clinician to give an overall assessment of the patients' clinical symptom

severity based on all of the outcome measures and their movement over the entirety of the study.

[187] The responder analysis for CGI-S at Week 4 demonstrates a clear dose response trend of

improvement for the L1-79 100 mg and 200 mg groups compared to placebo.

Vineland Adaptive Behavior Scales, Second Edition (VABS II)

[188] The change from Baseline in VABS II Standardized Socialization Score and the VABS II

Standardized Communication Score for L1-79 compared to placebo over time are displayed

graphically in Figure 12 and Figure 13, respectively.

Autism Diagnostic Observation Schedule, Second Edition (ADOS-2)

[189] The change from Screening in ADOS-2 Total Score, Restrictive and Repetitive Behavior

Total Score and Social Affect Total Score for L1-79 compared to placebo over time are displayed

graphically in Figure 14, Figure 15 and Figure 16. While not typically used as an outcome

measure, the change in ADOS-2 over a short period of time is consistent with open-label

preliminary study of longer duration previously presented.

[190] The consistency of this effect across multiple patients is suggested by the summary of the

change in ADOS-2 Total Score from Screening to Week 4 displayed in Figure 17.

Social Responsiveness Scale, Second Edition (SRS-2)

[191] The change from Baseline in SRS-2 Total T-score, SRS-2 DSM-5 Social, Communication

and Interaction T-score, SRS-2 Social Communication T-score, SRS-2 Social-Motivation T-score

and SRS-2 DSM-5 Restrictive and Repetitive Behavior T-score for L1-79 compared to placebo

over time are displayed graphically in Figure 18, Figure 19, Figure 20, Figure 21 and Figure 22,

respectively.

[192] The SRS-2 Total, SRS-2 DSM-5 Social Communication and Interaction, SRS-2 Social

Communication and SRS-2 Social Motivation T-scores improved by close to 8 points or more on

average in the L1-79 200 mg treated group (Figure 18, Figure 19, Figure 20 and Figure 21). The

Social Communication and Interaction scale are comprised of the DSM-5 criteria that make up the

social communication and social interaction deficits required for the diagnosis of ASD. Of

considerable interest is the finding that patients with baseline scores between 60 and 83 who

dropped by 8 points or more were likely to demonstrate categorical changes in clinical severity,

since the classifications of "Within Normal Limits" (below 60 T-score), "Mild Range" (60 to 65 T-

score), "Moderate Range" (66 to 75 T-score) and "Severe Range" (76 T-score or greater) of

symptom severity are defined within those T-score ranges.

[193] A summary of the change in SRS-2 Total T-score, SRS-2 DSM-5 Social, Communication

and Interaction T-score, SRS-2 Social Communication T-score, and SRS-2 Social-Motivation T-

score from baseline to Week 4 for each patient are displayed in Figure 23, Figure 24, Figure 25.

and Figure 26, respectively.

[194] For SRS-2 Total, SRS-2 DSM-5 Social Communication and Interaction, SRS-2 Social

Communication, and SRS-2 Social Motivation T-scores, both the magnitude and number of

responders observed were greater with L1-79 200 mg, compared to placebo (Figure 27, Figure

28, Figure 29 and Figure 30). For the SRS-2 Total T-Score, 3 patients had sufficient improvement

to change categories of severity in the L1-79 200 mg group, compared to 2 in both the L1-79 100

mg and placebo groups. For the SRS-2 Social Motivation T-score, 6 patients demonstrated

categorical changes in clinical severity in the L1-79 200 mg group, compared to 4 and 3 patients

in the 100mg and placebo groups, respectively. For the SRS-2 DSM-5 Social, Communication and

Interaction T-score, 4 patients had changes in severity categorization in the 200 mg group,

compared to 2 and 3 patients in the 100 mg and placebo groups, respectively. Similarly, for the

PCT/US2019/040581

SRS-2 Social Communication T-score, 4 patients demonstrated categorical improvement in the

200 mg group, compared to 3 in both the placebo and 100 mg groups.

[195] Responder analyses (defined as an improvement or no worsening) are presented for SRS-

2 Total T-score, SRS-2 DSM-5 Social, Communication and Interaction T-score, SRS-2 Social

Communication T-score, and SRS-2 Social-Motivation T-score at Week 4 or LOCF in Figure 27,

Figure 28, Figure 29, and Figure 30, respectively.

[196] Figures 27-30 demonstrate that the percent of responders (as defined by improvement or

no worsening were greater with L1-79 200 mg, compared to placebo; see Responder Definition

(section above).

Aberrant Behavior Checklist - Community (ABC-C)

[197] The change from Baseline in ABC-C Lethargy and Social Withdrawal Domain, ABC-C

Inappropriate Speech Domain, ABC-C Hyperactivity and Noncompliance Domain, ABC-C

Stereotypic Behavior Domain and ABC-C Irritability and Agitation Domain for L1-79 compared

to placebo over time are displayed graphically in Figure 31, Figure 32, Figure 33, Figure 34 and

Figure Figure 35, 35,respectively. respectively.

[198] A summary of the change in ABC-C Lethargy and Social Withdrawal Domain and ABC-

C Inappropriate Speech Domain from Baseline to Week 4 are displayed by patient in Figure 36

and Figure 37, respectively. While there was no separation between the 200 mg and placebo

groups in mean change from baseline scores, responder analyses did demonstrate a greater

tendency for patients to respond with improvement or no worsening of behaviors in the 200 mg

group compared to placebo, as shown in Figure 38 and Figure 39, respectively.

[199] As with the SRS-2, both the number of responders and magnitude of response for those

patients given L1-79 200 mg demonstrated a trend toward separation for the social and speech

domains of the ABC-C. Equally notable was the observation that 90 to 100% of patients with L1-

79 200 mg demonstrated stability or improvement of these domains, which are known to vary

significantly in intensity over shorter periods of time.

Repetitive Behavior Scale - Revised (RBS-R)

[200] The change from Baseline in RBS-R Total Score, RBS-R Restrictive Behavior, RBS-R

Ritualistic Behavior, RBS-R Sameness Behavior, RBS-R Compulsive Behavior, RBS-R

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Stereotypic Behavior, and RBS-R Self-injurious Behavior for L1-79 compared to placebo over

time is displayed graphically in Figure 40, Figure 41, Figure 42, Figure 43, Figure 44, Figure

45, and Figure 46.

[201] There were mean improvements in RBS-R Total Score as noted by a decrease in score for

L1-79 200 mg compared to placebo. (Figure 40).

[202] A summary of the change in RBS-R Total Score from Baseline to Week 4 is displayed by

patient in Figure 47. In addition, responder analyses (defined as an improvement or no worsening)

are presented for RBS-R Total Score at Week 4 or LOCF in Figure 48.

[203] Consistent with the ABC-C Lethargy and Social Withdrawal and ABC-C Inappropriate

Speech Domains, the RBS-R Total Score demonstrated a clear trend toward a greater magnitude

of reduction in restricted and repetitive behaviors with L1-79 200 mg compared to placebo. This

is further demonstrated by the responder analysis, which shows that 91% of patients treated with

L1-79 200 mg had improvement or stabilization of symptoms compared to 78% with placebo.

Overall Conclusions

[204] Currently available therapies for children and adults with ASD only target collateral

symptoms associated with the disorder (irritability, agitation, impulsivity, hyperactivity) and have

side effects that require monitoring metabolic parameters through blood tests. While phlebotomy

is unpleasant for most neurotypical children and adults, it is quite traumatic for those with extreme

sensory sensitivities who are unable to adequately communicate their fears. Indeed, it is a catch-

22 for these individuals, in that many of the aberrant behaviors they exhibit are likely due to an

inability to interact, communicate and connect with others. The potential to provide them with a

therapy that not only improves upon the core symptoms responsible for those behaviors, but to do

so without the need for invasive monitoring will go a long way to improve the quality of life for SO

these children.

[205] Results from Study HT 02-121 provide proof of concept that L1-79 appears to provide

benefit in treating the core symptoms of ASD. ASD is defined primarily as persistent deficits in

social communication and social interaction as well as restricted and repetitive behavior patterns,

interests or activities. Preliminary evidence from multiple independent assessments specifically

used to measure both the social and behavioral intensity of these core symptoms demonstrated

WO wo 2020/018292 PCT/US2019/040581

consistent trends that were repeatable across multiple instruments. While the limited size and short

duration of the study precluded any expectation or ability to demonstrate statistically significant

improvements in the outcome measures used, the agreement between the multiple measures

utilized in this study is very encouraging. In less than one month of treatment, the blinded

assessment by the clinicians involved in the study demonstrated a nearly one-point change in the

CGI-S compared to baseline. Similarly, patients receiving L1-79 improved by nearly one point

from baseline in ADOS-2 scores within the same time period.

[206] Previous experience (see Example 5) with open-label administration of L1-79 in 10

patients with autism clearly demonstrated that L1-79 has the potential to improve the core

symptoms of autism. In Study HT 02-121, multiple independent efficacy measures including the

CGI-S, ADOS-2, SRS-2, ABC-C and RBS-R demonstrated consistent improvements in the target

score social domains affected by ASD despite the short treatment period and small number of

patients. Over the 28-day treatment period, L1-79 was safe and well tolerated. At the conclusion

of the study, many parents wanted their children to continue receiving treatment with L1-79. As

a result, a roundtable videos were filmed at both clinical sites with the intent of allowing these

parents to speak directly with FDA about the impact of L1-79 on their children with autism.

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EXAMPLE 7 A Randomized, Double-Blind, Placebo-Controlled Adaptive Trial of L1-79 for the

Treatment of the Core Deficits in Social-Communication Function and Adolescents and

Adults with Autism Spectrum Disorder

[207] The completed Study HT 02-121 (Example 6) was a Phase II safety study of L1-79 for the

treatment of autism. Study HT 02-121 was a randomized double-blind, placebo-controlled two-

cohort, 4-week dose-escalation study that incorporated 2 open-label treatment groups to assess the

safety and efficacy of L1-79 100 mg and 200 mg TID in male patients between the ages of 13 and

21 years of age with autism. Results from Study HT 02-121 provide proof of concept that L1-79

appears to provide benefit in treating the core symptoms of ASD. Preliminary evidence from

multiple independent assessments specifically used to measure both the social and behavioral

intensity of these core symptoms demonstrated consistent trends that were repeatable across

multiple instruments. While the limited size and short duration of the study precluded any

expectation or ability to demonstrate statistically significant improvements in the outcome

measures used, the agreement between the multiple measures utilized in this study is very

encouraging. Following a review of the results from Study HT 02-121 the FDA allowed L1-79 to

begin registration trials leading toward a marketing approval and awarded the L1-79 IND 128673

a Fast Track designation.

[208] As a result, the inventors are proposing an adaptive trial approach in a Phase III setting that

will allow an expedient yet thorough approach to evaluating L1-79 as a therapy for treating the

defining core deficits in social communication and interaction. Based on the results from Study

HT 02-121 (Example 6), proposed are two Phase III randomized, double-blind, parallel group,

placebo controlled, clinical studies (Study 301 and Study 302) utilizing adaptive designs in order

to evaluate optimal inclusion criteria, sample size and outcome measures during the first segment

of Study 301 to quantify the safety and efficacy of L1-79 administered TID at doses of 200 or 300

mg to subjects with a diagnosis of autism based upon their Autism Diagnostic Observation

Schedule-2 (ADOS-2) results in a prospectively randomized and double-blind manner.

[209] Most of the clinical experience with L1-79 for the treatment of autism is with doses of 100-

200 mg TID. The study of Example 8 (also referred to herein as "Study 301") seeks to quantify

the safety and efficacy of L1-79 administered TID at doses of 200 or 300 mg to subjects with a

WO wo 2020/018292 PCT/US2019/040581

diagnosis of autism based upon their Autism Diagnostic Observation Schedule-2 (ADOS-2) results

in a prospectively randomized and double-blind manner.

[210] The dosage form is a capsule containing 100 mg of DL-a-methyl-para-tyrosine capsules DL--methyl-para-tyrosine capsules

(hereafter referred to as "L1-79"). The intended dosing regimen of L1-79 for Phase III clinical

studies (Study 301 and Study 302) is 2 or 3 capsules administered orally three times daily (TID).

Rationale for Study Design:

[211] ASD is a disorder marked by deficits in social interaction and the presence of restricted,

repetitive patterns of behavior, interests, or activities during childhood development (Swedo, S.

E., Baird, G., Cook, E. H., Happe', F. G., Harris, J. C., Kaufmann, W. E.,Wright, W.E., Wright,Harry, Harry,H. H.

(Eds.). (2013). Neurodevelopmental Disorders. In American Psychiatric Association. Diagnostic

and statistical manual of mental disorders (5th ed.). American Psychiatric Association.). Recent

literature has implicated peripheral and autonomic nervous system involvement in children and

adults with ASD (Baker, 2017; Fenning, 2017). As a result, electrodermal skin testing may serve

as an important biomarker for the population of patients more likely to respond to treatments

targeting the sympathetic nervous system.

Primary Objective(s):

[212] The primary objectives of the study are to evaluate the efficacy, safety, and tolerability of

L1-79 compared to placebo for the treatment of the core deficits in social communication and

interaction in adolescents and adults with autism spectrum disorder (ASD).

Secondary Objective(s):

[213] Secondary objectives of the study include the following:

[214] 1. 1.

[214] to to evaluate evaluate optimal optimal inclusion inclusion criteria, criteria, sample sample size, size, andand outcome outcome measures measures during during thethe first first

segment of this adaptive trial;

[215] 2. to evaluate the effect of L1-79 compared to placebo on reducing repetitive and restrictive

behaviors in ASD;

[216] 3. to evaluate the effect of L1-79 compared to placebo on reducing aberrant behaviors in

ASD, including hyperactivity, agitation, and irritability; and

56

WO wo 2020/018292 PCT/US2019/040581

[217] 4. to perform additional modeling of pharmacokinetics (PK) and pharmacodynamics (PD)

for L1-79 in a subset of subjects through timed intermittent sparse sampling.

Study Design:

[218] Eligible participants will be adolescents and adults between the ages of 12 and 21 years

who meet the Diagnostic and Statistical Manual of Mental Disorders-5th edition (DSM-5) criteria

for ASD, based upon clinician interview and assessment of ASD symptoms on the ADOS-2, and

a Clinical Global Impression of Severity (CGI-S) rating of 4 or greater (moderate or higher).

[219] Subjects will be randomized to placebo, L1-79 200 mg, or L1-79 300 mg three times daily

(TID) groups in a 1:2:2 ratio.

[220] Subjects in the 200 and 300 mg group will be started on 200 mg TID. Subjects in the 300

mg group mg groupwill willtitrate up to titrate up 300 mg TID to 300 mg after 7 days.7 days. TID after

[221] Screening assessments will include the ADOS-2, a review of ASD criteria from the DSM-

5 and the CGI-S.

[222] An adaptive design will be used with a proposed interim analysis being used to make

decisions on the following criteria; potential endpoints for the primary outcome, modification of

the defined study population, and estimated sample size. The data monitoring committee and

independent statistical group will implement the interim analysis and make recommendations to

the sponsor regarding proposed changes in the study based on predefined criteria. The sample size

recalculation will depend on the primary outcome and potential study populations that will be

determined at the interim analysis. The sample size recalculation will be based on blinded data.

[223] A schedule of events is provided in Table 30.

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Table 30 Schedule of Events

Treatment TreatmentPersing Annia Follow-up Fellow-up Servening Screening Visit 3 Flate y Visit : Visit 3 Visit 4 Visit 45 Weir Valt S $ Not 2 4 The $ Evaluation Days will to .3 -3 Day 34 14 is = $$ Day Day 84*** Day 84 * 33 Day 120 * = 3 Day a & Day 28a 2& 33 60 * 3 Day to $

Informad Informed Consent X Contexia X

laeE Demographies Demographics Medical Mistory

Physical History 35 X X X % X" XP XM X² X Visa Signed X X X X X x X % X X x Cline UnitedDrug DrugSavena Screen* X 3 M Pregmancy Prequancy Tear Text X X (PK) (PK)w 3y is is exper past done dose X PX Randeen PK Random xxxxxx: (known issue after doss) after done) X" In an X X Laboratory Tears X K X X Randomoxation X 12-Leas ECG 12-Lead ECC X X X X Drug Administration X X X X X Drug Accountability X X X X Address Event (AB) Advene Exect (AE) Assessment Assessment X X R X X X X Athalization Prior/Canomitant Midication Assessment X X % % X X X X X X ADOS-2" X X 30 CGI-S CGL-S X X X X X x CGF-C X X CSRC X X 3 X X VABS-30 VABS-39 X X X X X SRS-3 SRS-2 X X X X X X

Table 30 Schedule of Events Treatment Period Follow-up Fallom-up Servening Screening Visit 1 ) Visit Flow $$ View S$ Visit Flow 43 Visit $ Visit &# Flor UW $ Evaluation Days 30 is to .3 -1 Day 9 $ Day Day 14 H ** 33 Day No 3 Day Sil SW **33 Day $4 * 3 # 8. Day Day 120 120* *3 $ Day 3 ABC-C X X X X X KBS-R RBS-R X X X X X N PSI X X X SSP SSP X X WASTE WASHI X DSM-5 for DSMA for ASD ASD X X Skin Receivity ReactivityTesting Testing(SRT) (SRT) X X M X

ABC-C = Aberrant Behavior Checklist-Community: Checklist-Community; ADOS-2 = Autism Diagnostic Observation Scale-2; ASD = autism spectrum disorder; CGI-C = Clinical Global Impression of Change; CGI-S = Clinical Global Impression of Severity; DSM-5 = Diagnostic and Statistical Manual of Mental Disorders-5th edition; PSI = Parenting Stress Index; RBS-R = Repetitive Behavior Scale-Revised; SAS = Spence Anxiety Scale; SSP = Spence Anxiety Scale; SRS-2 = Social Responsiveness Scale-2; VABS-3 = Vineland Adaptive Behavior Scales - 3rd Edition; WASI-2 = Weschler Abbreviated Scales of Intelligence 1 1 Includes Includes aa review review of of previous/ongoing previous/ongoing medications medications 2 Complete examination, including assessments of the skin, head, eyes, ears, nose, throat, neck, thyroid, lungs, heart, abdomen, lymph nodes, extremities, and body weight 3 Height will be measured at screening only. 4 Partial examination, to update findings from the examination performed at screening

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5 Includes respiratory rate, oral temperature, sitting and standing orthostatic blood pressure and pulse 6 6 Includes amphetamines, barbiturates, cocaine metabolites, opiates, benzodiazepines, cannabinoids, and cotinine 7 Only for females who are post-menarche 8 8 Subject will will only only have have random random PK PK done done for for one one of of Visits Visits 2, 2, 3, 3, or or 4. 4. Subject 9 Includes serum chemistry, hematology (including coagulation), and urinalysis

ADOS-2 Will Will be be videotaped videotaped for for potential potential use use as as an an additional additional review review for for Reciprocal Reciprocal Social Social Interaction Interaction ADOS-2 by Blinded reviewer 11 11 Adaptive Behavior Adaptive BehaviorComposite domains Composite only only domains

[224] Inclusion Criteria:

1. Subjects must be male or female adolescents or adults up to age 21

2. Subjects must be between the ages of 13 and 21 years of age

3. Post menarche females must be on birth control if appropriate.

4. Diagnosis of ASD based upon an assessment tool that utilizes the DSM-5 criteria [e.g., Autism

Symptom Rating Scale (ASRS), Childhood Autism Rating Scale-2 (CARS2), or Autism

Diagnostic Interview-Revised (ADI-R)], and confirmed with the ADOS-2, with a CGI-S score of

4 or greater.

5. Subject must be stable on no more than one concomitant medication, and no planned changes

in psychosocial interventions during the trial

6. Subjects and caregiver must be willing and able to participate in the testing procedures sufficient

to obtain valid scores on the tests used herein.

7. Subjects must have a caregiver who has known the them for over a year, spends at least 10 hours

per week with them, and is willing to accompany them to each appointment.

8. Subjects must, in the opinion of the Investigator, be sufficiently tolerant and capable of

complying with the requirements of this trial. For example, patients who will not tolerate blood

draws or ECG are not qualified candidates for this study.

9. Subjects must be able to swallow capsules

10. Subjects and their care givers must be willing to sign informed consent or to have informed

consent provided by their legal guardians or proxies. All subjects <18 years old or those unable

to care for themselves must have the caregiver's consent.

WO wo 2020/018292 PCT/US2019/040581

[225] Exclusion Criteria:

1. Sexually active males and females

2. Uncontrolled intercurrent illness including, but not limited to, ongoing or active infection,

symptomatic cardio-vascular disease, hepatic disease, renal disease, skeletomuscular disease, HIV,

HCVA, HBV, or psychiatric illness/social situations that would limit compliance with study

requirements.

3. Any disease that requires treatment with immunosuppressive drugs

4. A diagnosis of Fragile-X syndrome, Rett syndrome, or other neurological disorder that could be

the basis for the subjects autistic symptoms (e.g., congenital or acquired brain injury, brain

malformations, stroke, neurogenetic or metabolic disorder).

5. A DSM-5 diagnosis of schizophrenia, schizoaffective disorder, alcohol use disorder or ADHD,

Current or lifetime diagnosis of severe psychiatric disorder (e.g., bipolar disorder, etc.);

6. The Presence of any active chronic medical problem including, but not limited to uncontrolled

seizure disorder, heart disease, cancer, asthma, genetic disease, or any disease or syndrome that

requires continuous drug therapy.

7. Subjects requiring more than 1 medication for the treatment of autism, or who have not been

weaned weaned to totheir theirlowest tolerable lowest dose dose tolerable of medication. of medication.

8. Subjects with any disease that requires treatment with immunosuppressive drugs.

9. The presences of out of range hepatic or renal function tests or other unexplained abnormal

laboratory value that is deemed clinically significant by the Investigator.

10. Any subject or caregiver who is unwilling or unable to give informed consent.

Study Population:

[226] 350 patients are planned.

Test Product, Dose, and Mode of Administration:

[227] D-L alpha-methyl-tyrosine (L1-79) encapsulated as 100 and 200 mg. Two dose arms will

be tested as 200 mg TID (one 200 mg dose combined with placebo) and 300 mg TID (one 200 mg

capsule and one 100 mg capsule)

WO wo 2020/018292 PCT/US2019/040581

Duration of Treatment:

[228] 12 weeks (84 days)

Efficacy Assessments:

[229] Baseline assessments not included in the measurement of efficacy include the Weschler

Abbreviated Scales of Intelligence (WASI-2) and the Spence Anxiety Scale (SAS). Electrodermal

testing will be performed at baseline to determine if greater autonomic nervous system variability

correlates with response to treatment. (See Study Rationale). The primary outcome measure will

be the change from baseline at Week 12 on a 5-factor composite measure comprising the

Socialization (SOC), Communication (COM), and Daily Living Skills (DLS) domains from the

Adaptive Behavior Composite (ABC) of the Vineland Adaptive Behavior Scales - 3rd Edition

(VABS-3) and the Socialization, Communication and Interaction (SCI) and Restricted Interests

and Repetitive Behavior (RRB) subscales of the Social Responsiveness Scale-2 (SRS-2). The

primary secondary outcome measure will be based on the change from baseline CGI-S at Week

12.

[230] Additional secondary outcome measures will be assessed in a gated fashion (to be

determined during the interim analysis) for the change from baseline scores at Week 12 for:

1. Aberrant Behavior Checklist-Community (ABC-C) Social Withdrawal/Lethargy and

Inappropriate Speech Domains

2. ABC-C, Irritability, Hyperactivity, and Stereotypy Domains

3. Repetitive Behavior Scale-Revised (RBS-R)

4. Parenting Stress Index (PSI) Short Form

5. Sensory Profile - Short Form (SSP)

6. 6. Spence Anxiety Scale (SAS)

7. ADOS-2 Total Score* (ADOS-2 will be videotaped at screening and at end of study for

potential use in blinded review of reciprocal interaction.)

Safety Assessments:

[231] Safety assessments include physical examination, orthostatic measurements of blood

pressure and pulse, standard hematology and clinical chemistry assessments, concomitant

medication use, urinalysis, ECGs, and spontaneously reported adverse events.

WO wo 2020/018292 PCT/US2019/040581

PK Assessments:

[232] In order to gain a greater understanding of the PK and PD of L1-79 in patients with ASD,

sparse sampling will be utilized to assist in modeling PK parameters and comparing the same to

data obtained from more thorough studies done in young healthy adults. Sparse sampling will limit

the number of phlebotomy procedures, an important consideration in children with ASD who are

typically much more traumatized by blood drawing procedures than peers without ASD.

Statistical Methods:

[233] The adaptive design will assess three components of the study; the outcome variable, a

potential modification of the inclusion criteria, and a sample size recalculation based on the

decisions made at the time of the interim analysis. The modification of the inclusion criteria will

be based on an evaluation of potential endophenotypes for ASD of baseline severity of the

following characteristics known to directly influence treatment and intervention strategies for

people with ASD: 1) Severity of anxiety based on assessment by the Spence Anxiety Scale, IQ as

assessed by the Weschler Abbreviated Scale of Intelligence and disruptive behavior symptom

severity, based on the ABC-C irritability and hyperactivity subscales. In addition, skin reactivity

testing to determine autonomic sensitivity will be used as an exploratory biomarker for responder

phenotype (See Study Rationale for justification for these subgroups). The choice of primary

outcome(s) will be based on a conditional power analysis of the 5- factor composite measure

comprising the Socialization (SOC), Communication (COM), and Daily Living Skills (DLS)

domains from the Adaptive Behavior Composite (ABC) of the Vineland Adaptive Behavior Scales

- 3rd Edition (VABS-3) and the Socialization, Communication and Interaction (SCI) and

Restricted Interests and Repetitive Behavior (RRB) subscales of the Social Responsiveness Scale-

2 (SRS-2). After selection of the outcomes and inclusion criteria, the sample size will be

recalculated based on blinded data.

[234] It will be appreciated by those skilled in the art that changes could be made to the

embodiments described above without departing from the broad inventive concept thereof. It is is

understood, therefore, that this invention is not limited to the particular embodiments disclosed,

but it is intended to cover modifications that are within the spirit and scope of the invention, as

defined by the appended claims.

27304350.1:DCC-7/8/2025

[235] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[236] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or 2019308501

admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

62a

Claims (5)

27304350.1:DCC-7/8/2025 WHAT IS CLAIMED IS:

1. A method for providing a plasma concentration of a therapeutic drug for a long term for treating an autism in a subject in need thereof, the method comprising administering to said subject said therapeutic drug at a concentration ranging from about 50 mg (w/w) to about 1500 mg (w/w) daily, wherein said therapeutic drug is α-methyl-DL-tyrosine, wherein said plasma concentration ranges from about 500 ng/ml to 5000 ng/ml, and wherein said term is at least 1 week. 2019308501

2. The method of claim 1, wherein the concentration of said therapeutic drug is about 90 mg (w/w/).

3. The method of claim 1, wherein the concentration of said therapeutic drug is about 100 mg (w/w/).

4. The method of claim 1, wherein the concentration of said therapeutic drug is about 200 mg (w/w/).

5. The method of claim 1, wherein the concentration of said therapeutic drug is about 250 mg (w/w/).

6. The method of claim 1, wherein the concentration of said therapeutic drug is about 300 mg (w/w/).

7. The method of claim 1, wherein the concentration of said therapeutic drug is about 350 mg (w/w/).

8. The method of any one of claims 1 to 7, wherein said therapeutic drug is administered in a plurality of divided doses.

9. The method of any one of claims 1 to 8, wherein said therapeutic drug is administered for a duration ranging from about 1 week to about 4 weeks or more.

10. The method of any one of claims 1 to 8, wherein said plasma concentration ranges from about 800 ng/ml to 2500 ng/ml

11. The method of any one of claims 1 to 8, wherein said plasma concentration ranges from about 1400 ng/ml to 1800 ng/ml

27304350.1:DCC-7/8/2025

12. A method for treating an autism associated clinical trait in a subject in need thereof, the method comprising administering to said subject a composition comprising a therapeutically effective amount of α-methyl-DL-tyrosine, thereby treating said autism associated clinical trait in said subject.

13. The method of claim 12, wherein said clinical trait is a deficit in social communication, a deficit in social interaction, a deficit in social motivation, lethargy and social withdrawal, 2019308501

inappropriate speech, hyperactivity, stereotypic behavior, irritability and agitation, restrictive behavior, repetitive behavior, ritualistic behavior, sameness behavior, compulsive behavior, self- injurious behavior or a combination thereof.

14. The method of claim 12, wherein said clinical trait meets the requirements of Diagnostic and Statistical Manual of Mental Disorders – V (DSM-V) criteria.

15. The method of claim 12, wherein said clinical trait is assessed based upon a change from a baseline in one or more psychometric tests.

16. The method of claim 15, wherein said psychometric test is based on clinical global impression (CGI) rating scale, Vineland adoptive behavior scale (VABS), autism diagnostic observation schedule (ADOS), social responsiveness scale (SRS), aberrant behavior checklist – community (ABC-C), repetitive behavior scale (RBS), Conners parent rating scale (CPRS), or a combination thereof.

17. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount ranging from about 50 mg (w/w) to about 1000 mg (w/w).

18. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about 90 mg (w/w/).

19. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about100 mg (w/w/).

20. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about 200 mg (w/w/).

21. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about 250 mg (w/w/).

27304350.1:DCC-7/8/2025

22. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about 300 mg (w/w/).

23. The method of any one of claims 12 to 16, wherein the composition comprises α-methyl- DL-tyrosine in an amount of about 350 mg (w/w/).

24. The method of any one of claims 12 to 23, wherein the composition is administered in a plurality of divided doses. 2019308501

25. The method of any one of claims 12 to 24, wherein the composition is administered three times per day.

26. The method of any one of claims 12 to 25, wherein the composition is administered for at least 1 week.

27. The method of any one of claims 12 to 25, wherein the composition is administered for a duration ranging from about 1 week to about 4 weeks.

28. A method for providing a plasma concentration of a therapeutic drug for a long term for treating an autism in a subject in need thereof, the method comprising administering to said subject said therapeutic drug at a concentration ranging from about 50 mg (w/w) to about 500 mg (w/w) daily, wherein said therapeutic drug is α-methyl-DL-tyrosine, wherein said plasma concentration ranges from about 500 ng/ml to 5000 ng/ml, and wherein said term is at least 1 week.

CPRS All Pts

45 42

40 39

35 35 33

30 29

25 25 20 20 18

15

10

5 5

0 LR RS WC 11/1/2015

11/29/2015

Figure 1

SUBSTITUTE SHEET (RULE 26)