PTI-125 for Mild-to-moderate Alzheimer's Disease Patients

• Sponsor: Cassava Sciences, Inc. (Industry)
• Overall Status: Completed
• CT.gov ID: NCT04079803
• Collaborator: National Institute on Aging (NIA) (NIH)
• Enrollment: 64
• Locations: 9
• Arms: 3
• Actual Duration (Months): 6.7
• Patients Per Site: 7.1
• Patients Per Site Per Month: 1.1

Study Details

Study Description

Brief Summary

This is a Phase 2b, Randomized, Double-blind, Placebo-controlled, multiple dose study of PTI-125 in mild-to-moderate Alzheimer's disease patients.

Condition or Disease	Intervention/Treatment	Phase
Alzheimer Disease	Drug: Placebo oral tablet; Drug: Simufilam 100 mg tablet; Drug: Simufilam 50 mg oral tablet	Phase 2

Detailed Description

This is a Phase 2b, Randomized, Double-blind, Placebo-controlled, multiple dose study of PTI-125 in mild-to-moderate Alzheimer's disease patients. A total of sixty (60) patients will be enrolled in the study. Patients will receive Placebo, 50 mg or 100 mg b.i.d. of PTI-125. The objective of this study are to investigate the safety, and biomarkers of PTI-125 following 28-day repeat oral administration.

Study Design

• Study Type: Interventional
• Actual Enrollment: 64 participants
• Allocation: Randomized
• Intervention Model: Parallel Assignment
• Intervention Model Description: Approximately sixty (60) patients will be enrolled into the study and randomized to one of three cohorts. Cohorts will receive placebo or PTI-125 at 50 or 100 mg b.i.d. (n=20 per group)Approximately sixty (60) patients will be enrolled into the study and randomized to one of three cohorts. Cohorts will receive placebo or PTI-125 at 50 or 100 mg b.i.d. (n=20 per group)
• Masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
• Masking Description: The sponsor, participant, care provider, investigator including sub-investigators and outcomes assessors will be blinded to throughout the study which includes using an Integrated Web Response System (IWRS) and electronic data capture (EDC) to ensure blinding during the study.
• Primary Purpose: Treatment
• Official Title: A Phase 2b, Randomized, Double-blind, Placebo-controlled, Multiple Dose, Biomarker and Safety Study of PTI-125 in Mild-to-moderate Alzheimer's Disease Patients
• Actual Study Start Date: Sep 9, 2019
• Actual Primary Completion Date: Mar 31, 2020
• Actual Study Completion Date: Mar 31, 2020

Arms and Interventions

Arm	Intervention/Treatment
Placebo Comparator: Placebo Cohort Subjects administered placebo oral tablets twice daily (BID)	Drug: Placebo oral tablet; Oral placebo tablet
Experimental: Simufilam (PTI-125) 100 mg tablets Cohort Subjects administered simufilam (PTI-125) 100 mg oral tablets twice daily (BID)	Drug: Simufilam 50 mg oral tablet; Simufilam 50 mg oral tablet; Other Names: PTI-125
Experimental: Simufilam (PTI-125) 50 mg tablets Cohort Subjects administered simufilam (PTI-125) 50 mg oral tablets twice daily (BID)	Drug: Simufilam 100 mg tablet; Simufilam 100 mg oral tablet; Other Names: PTI-125

Outcome Measures

Primary Outcome Measures

1. Change From Baseline in CSF Abeta42 [Screening to Day 28]

   Change from Baseline (screening sample) to Day 28 in cerebrospinal fluid levels of Amyloid beta42

2. Change From Baseline in CSF Total Tau. [Screening to Day 28]

   Change from Baseline (screening sample) to Day 28 in cerebrospinal fluid total tau.

3. Change From Baseline in CSF P-tau181 [Screening to Day 28]

   Change from Baseline (screening) to Day 28 in cerebrospinal fluid P-tau181

4. Change From Baseline in CSF Neurogranin [Screening to Day 28]

   Change from Baseline (screening) to Day 28 in cerebrospinal fluid neurogranin

5. Change From Baseline in CSF Neurofilament Light Chain [Screening to Day 28]

   Change from Baseline (screening) to Day 28 in cerebrospinal fluid neurofilament light chain

6. Change From Baseline in CSF YKL-40 [Screening to Day 28]

   Change from Baseline (screening) in cerebrospinal fluid YKL-40

Secondary Outcome Measures

1. Paired Associates Learning Test [Day 1 to Day 28]

   Cognitive test assessing episodic memory. Boxes are displayed on the screen and are "opened" in a randomized order. One or more of them contains a pattern. The patterns are then displayed in the middle of the screen, one at a time and the participant must select the box in which the pattern was originally located. If the participant makes an error, the boxes are opened in sequence again to remind the participant of the locations of the patterns. The number of boxes increases progressively to a total of 8.

2. Spatial Working Memory Test [Day 1 to Day 28]

   Cognitive assessment of spatial working memory: A number of colored squares (boxes) are shown on the screen. By selecting the boxes and using a process of elimination, the subject should find one yellow 'token' in each of a number of boxes and use them to fill up an empty column on the right-hand side of the screen. The number of boxes is gradually increased to a total of 8 for the subjects to search. The colors and positions of the boxes are changed from trial to trial to discourage stereotyped search strategies.

3. CSF IL-6, sTREM2, HMGB1, Albumin, IgG [Screening to Day 28]

   Change from Baseline (screening sample) to Day 28 in secondary CSF biomarkers of neuroinflammation and blood-brain barrier integrity

Other Outcome Measures

1. Target Engagement Assays: Change From Baseline in Filamin A (FLNA) Linkages to alpha7 Nicotinic Acetylcholine Receptor (alpha7nAChR) and Toll-like Receptor 4 (TLR4) in Subject Lymphocytes [Day 1 to Day 28]

   FLNA linkages to these two receptors were assessed by densitometric quantitation of immunoblot bands of each receptor (detected by a specific antibody) in anti-FLNA precipitates. The measure is noted as a ratio to total FLNA.

2. Plasma P-tau181 [Day 1 to Day 28]

   Percent change in plasma P-tau181

3. Percent Change From Baseline in SavaDx, a Novel Plasma Biomarker [Day 1 to Day 28]

   SavaDx is a novel plasma biomarker

Eligibility Criteria

Criteria

• Ages Eligible for Study: 50 Years to 85 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No

Inclusion Criteria:

• Ages >= 50 and <= 85 years

• Informed consent form (ICF) signed by the subject or legally acceptable representative.

• Clinical diagnosis of dementia due to possible or probable Alzheimer's disease

• Mini-Mental State Examination score >= 16 and <= 26 at screening

• If female, postmenopausal for at least 1 year

• Patient living at home, senior residential setting, or an institutional setting without the need for continuous (i.e. 24-h) nursing care

• General health status acceptable for participation in the study

• Fluency (oral and written) in English or Spanish

• If receiving memantine, rivastigmine, galantamine or an AChEI, receiving a stable dose for at least 3 months. If receiving donepezil, any dose lower than 23 mg once daily.

• The patient is a non-smoker for at least 3 years.

• The patient or legal representative must agree to comply with the drawing of blood samples and with a lumbar puncture and the drawing of cerebrospinal fluid samples.

• The patient has a ratio of total tau/Aβ42 in cerebrospinal fluid >= 0.28.

• Patient has a caregiver or legal representative responsible for administering the drug and recording the time.

Exclusion Criteria:

• Exposure to an experimental drug, experimental biologic or experimental medical device within the longer of 5 half-lives or 3 months before screening

• Enrollment in the previous PTI-125 trial

• A medical condition that would interfere with a lumbar puncture

• Residence in a skilled nursing facility and requiring 24 h care.

• Clinically significant laboratory test results

• Clinically significant untreated hypothyroidism

• Insufficiently controlled diabetes mellitus

• Renal insufficiency (serum creatinine > ULN)

• Malignant tumor within 3 years before screening (except squamous and basal cell carcinoma or cervical carcinoma in situ or localized prostate cancer or localized stage 1 bladder cancer)

• History of ischemic colitis or ischemic enterocolitis

• Unstable medical condition that is clinically significant in the judgment of the investigator

• Alanine transaminase (ALT) or aspartate transaminase (AST) > ULN or total bilirubin > ULN.

• History of myocardial infarction or unstable angina within 6 months before screening

• History of more than 1 myocardial infarction within 5 years before screening

• Clinically significant cardiac arrhythmia (including atrial fibrillation), cardiomyopathy, or cardiac conduction defect (patients with a pacemaker are acceptable)

• Symptomatic hypotension, or uncontrolled hypertension

• Clinically significant abnormality on screening electrocardiogram (ECG), including but not necessarily limited to a confirmed corrected QT interval value >= 450 msec for males or >= 470 msec for females.

• Stroke within 18 months before screening, or history of a stroke concomitant with onset of dementia

• History of brain tumor or other clinically significant space-occupying lesion on CT or MRI

• Head trauma with clinically significant loss of consciousness within 12 months before screening or concurrent with the onset of dementia

• Onset of dementia secondary to cardiac arrest, surgery with general anesthesia, or resuscitation

• Specific degenerative Central Nervous System disease diagnosis other than Alzheimer's disease (eg, Huntington's disease, Creutzfeld-Jacob disease, Down's syndrome, Frontotemporal Dementia, Parkinson's disease)

• Wernicke's encephalopathy

• Active acute or chronic Central Nervous System infection

• Donepezil 23 mg quaque die currently or within 3 months prior to randomization

• Discontinued AChEI < 30 days prior to randomization

• Antipsychotics; low doses are allowed only if the subject has received a stable dose for at least 3 months before randomization

• Tricyclic antidepressants and monoamine oxidase inhibitors

• Anxiolytics or sedative-hypnotics, including barbiturates (unless given in low doses for benign tremor); low doses of benzodiazepines and zolpidem are allowed

• Immunosuppressants, including systemic corticosteroids, if taken in clinically immunosuppressive doses (Steroid use for allergy or other inflammation is permitted.)

• Antiepileptic medications if taken for control of seizures

• Chronic intake of opioid-containing analgesics

• Sedating H1 antihistamines

• Nicotine therapy (all dosage forms including a patch), varenicline (Chantix), or similar therapeutic agent within 30 days before screening

• Clinically significant illness within 30 days of enrollment

• History of significant neurological, hepatic, renal, endocrine, cardiovascular, gastrointestinal, pulmonary, or metabolic disease

• Positive serum hepatitis B surface antigen (HBsAg) or positive hepatitis C virus HCV antibody test at screening

• Positive HIV test at screening

• Positive urine drug test at screening

• Loss of a significant volume of blood (> 450 mL) within 4 weeks prior to the study

• Suicidality on C-SSRS at screening

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Cognitive Clinical Trials	Gilbert	Arizona	United States	85296
2	Cognitive Clinical Trials	Surprise	Arizona	United States	85374
3	Optimus U	Miami	Florida	United States	33125
4	IMIC, Inc.	Palmetto Bay	Florida	United States	33157
5	Cognitive Clinical Trials	Bellevue	Nebraska	United States	68005
6	Cognitive Clinical Trials	Omaha	Nebraska	United States	68116
7	Advanced Memory Research Institute	Toms River	New Jersey	United States	08755
8	Centex Studies, Inc.	Houston	Texas	United States	77058
9	Centex Studies, Inc.	McAllen	Texas	United States	78504

Sponsors and Collaborators

• Cassava Sciences, Inc.
• National Institute on Aging (NIA)

Investigators

• Study Director: Lindsay Burns, PhD, Cassava Sciences, Inc.

Study Documents (Full-Text)

• Study Protocol - Jun 28, 2019
• Statistical Analysis Plan - Feb 12, 2020
• Informed Consent Form - Jul 10, 2019

More Information

Additional Information:

• Altered filamin A enables amyloid beta-induced tau hyperphosphorylation and neuroinflammation in Alzheimer's disease
• PTI-125 and Alzheimer's Publications

Publications

None provided.

• Responsible Party: Cassava Sciences, Inc.
• ClinicalTrials.gov Identifier: NCT04079803
• Other Study ID Numbers: PTI-125-02; R44AG060878
• First Posted: Sep 6, 2019
• Last Update Posted: Sep 29, 2021
• Last Verified: Sep 1, 2021

• Individual Participant Data (IPD) Sharing Statement: No
• Plan to Share IPD: No
• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Additional relevant MeSH terms: Alzheimer Disease

Study Results

Participant Flow

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Subjects administered placebo oral tablets twice daily (BID)	Subjects administered simufilam 100 mg oral tablets twice daily (BID)	Subjects administered simufilam 50 mg oral tablets twice daily (BID)
Period Title: Overall Study
STARTED	22	21	21
COMPLETED	22	21	20
NOT COMPLETED	0	0	1

Baseline Characteristics

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort	Total
Arm/Group Description	Subjects administered matching placebo tablets twice daily for 28 days.	Subjects administered 100 mg simufilam tablets twice daily for 28 days.	Subjects administered 50 mg simufilam tablets twice daily for 28 days.	Total of all reporting groups
Overall Participants	22	21	21	64
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%	0 0%
Between 18 and 65 years	4 18.2%	9 42.9%	6 28.6%	19 29.7%
>=65 years	18 81.8%	12 57.1%	15 71.4%	45 70.3%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	71.3 (6.68)	69.3 (5.47)	67.1 (8.76)	69.2 (6.97)
Sex: Female, Male (Count of Participants)
Female	11 50%	12 57.1%	12 57.1%	35 54.7%
Male	11 50%	9 42.9%	9 42.9%	29 45.3%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	9 40.9%	11 52.4%	11 52.4%	31 48.4%
Not Hispanic or Latino	13 59.1%	10 47.6%	10 47.6%	33 51.6%
Unknown or Not Reported	0 0%	0 0%	0 0%	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%	0 0%	0 0%	0 0%
Asian	1 4.5%	0 0%	0 0%	1 1.6%
Native Hawaiian or Other Pacific Islander	0 0%	0 0%	0 0%	0 0%
Black or African American	2 9.1%	2 9.5%	4 19%	8 12.5%
White	19 86.4%	19 90.5%	17 81%	55 85.9%
More than one race	0 0%	0 0%	0 0%	0 0%
Unknown or Not Reported	0 0%	0 0%	0 0%	0 0%
CSF total tau/Aβ42 ratio (ratio) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [ratio]	1.20 (0.55)	1.08 (0.50)	1.17 (0.58)	1.15 (0.54)
Mini-Mental State Exam (MMSE) (units on a scale) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [units on a scale]	23.1 (2.78)	23.0 (2.66)	22.7 (2.67)	22.9 (2.70)
Taking cholinesterase inhibitor or memantine (Count of Participants)
Count of Participants [Participants]	8 36.4%	7 33.3%	5 23.8%	20 31.3%
Heterozygous APOE4 (Count of Participants)
Count of Participants [Participants]	12 54.5%	14 66.7%	9 42.9%	35 54.7%
Homozygous APOE4 (Count of Participants)
Count of Participants [Participants]	1 4.5%	1 4.8%	3 14.3%	5 7.8%
Paired Associates Learning total errors (errors) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [errors]	35.5 (19.65)	31.0 (20.74)	36.1 (18.76)	34.2 (19.7)
Spatial Working Memory total errors (errors) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [errors]	19.0 (7.49)	22.1 (5.88)	22.3 (6.64)	21.1 (6.67)
CSF Aβ42 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	125 (152)	117 (51.4)	108 (54.8)	111 (86.1)
CSF total tau (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	104 (32)	106 (27.9)	101 (17.6)	103.7 (25.8)
CSF P-tau181 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	28.5 (0.73)	29.7 (1.5)	29.0 (1.0)	29.1 (1.3)
CSF neurogranin (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	1200 (365)	1551 (751)	1352 (614)	1368 (577)
CSF Neurofilament Light Chain (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	161 (42.8)	219 (95.3)	181 (64.4)	187 (67.5)
CSF YKL-40 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	206 (29.5)	203 (22.7)	194 (26.0)	201 (26.1)
CSF IL-6 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	32.5 (1.2)	33.6 (1.8)	33.6 (1.7)	33.2 (1.6)
CSF sTREM2 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	878 (435)	861 (421)	882 (476)	874 (444)
CSF HMGB1 (pg/mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [pg/mL]	424 (48.0)	446 (67.3)	454 (70.6)	441 (62.0)
CSF/plasma albumin ratio (ratio) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [ratio]	0.24 (0.03)	0.25 (0.08)	0.25 (0.05)	0.25 (0.05)
CSF/plasma IgG ratio (ratio) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [ratio]	0.200 (0.07)	0.217 (0.11)	0.227 (0.07)	0.215 (0.08)
Lymphocyte filamin A - α7nAChR, Ratio to total filamin A (ratio) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [ratio]	0.59 (0.10)	0.69 (0.11)	0.66 (0.12)	0.65 (0.11)
Lymphocyte filamin A - TLR4, Ratio to total filamin A (ratio) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [ratio]	0.55 (0.10)	0.60 (0.07)	0.58 (0.11)	0.58 (0.09)

Outcome Measures

1. Primary Outcome

Title	Change From Baseline in CSF Abeta42
Description	Change from Baseline (screening sample) to Day 28 in cerebrospinal fluid levels of Amyloid beta42
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo tablets BID for 28 days	simufilam 100 mg tablets BID for 28 days	simufilam 50 mg tablets BID for 28 days
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	4.8 (30.9)	12.5 (11.9)	16.2 (21.1)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.087
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P=0.0088 and CFB was 13.4. In percent change from baseline with these two subjects removed, P=0.004.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.01
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.0006 and CFB was 16.9. In percent change from baseline with this subject removed, P=0.0004.

2. Primary Outcome

Title	Change From Baseline in CSF Total Tau.
Description	Change from Baseline (screening sample) to Day 28 in cerebrospinal fluid total tau.
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125) 100 mg Tablets Cohort	Simufilam (PTI-125) 50 mg Tablets Cohort
Arm/Group Description	Placebo oral tablets administered twice daily (BID)	Simufilam (PTI-125) 100 mg oral tablets administered twice daily (BID)	SImufilam (PTI-125) 50 mg oral tablets administered twice daily (BID)
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	-3.2 (14.8)	-18.7 (10.4)	-14.6 (9.6)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	<0.0001
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P<0.0001 and CFB was -19.8.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0012
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.001 and CFB was -14.9.

3. Primary Outcome

Title	Change From Baseline in CSF P-tau181
Description	Change from Baseline (screening) to Day 28 in cerebrospinal fluid P-tau181
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo tablets BID for 28 days	simufilam 100 mg tablets BID for 28 days	simufilam 50 mg tablets BID for 28 days
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	-0.63 (1.8)	-3.1 (1.7)	-2.4 (1.6)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.005
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P=0.003 and CFB was -3.2.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.002
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.003 and CFB was -2.5.

4. Primary Outcome

Title	Change From Baseline in CSF Neurogranin
Description	Change from Baseline (screening) to Day 28 in cerebrospinal fluid neurogranin
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo tablets BID for 28 days	simufilam 100 mg tablets BID for 28 days	simufilam 50 mg tablets BID for 28 days
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	-50.5 (434)	-648 (491)	-527 (361)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0002
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P=0.0002 and CFB was -681.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0005
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.0005 and CFB was -527.

5. Primary Outcome

Title	Change From Baseline in CSF Neurofilament Light Chain
Description	Change from Baseline (screening) to Day 28 in cerebrospinal fluid neurofilament light chain
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo tablets BID for 28 days	simufilam 100 mg tablets BID for 28 days	simufilam 50 mg tablets BID for 28 days
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	-10.0 (45)	-76.3 (50.6)	-49.7 (35.5)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0003
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P=0.0002 and CFB was -78.5.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0058
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.0008 and CFB was -51.0.

6. Primary Outcome

Title	Change From Baseline in CSF YKL-40
Description	Change from Baseline (screening) in cerebrospinal fluid YKL-40
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample. This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo tablets BID for 28 days	simufilam 100 mg tablets BID for 28 days	simufilam 50 mg tablets BID for 28 days
Measure Participants	22	21	19
Mean (Standard Deviation) [pg/mL]	-0.96 (24.2)	-22.3 (11.7)	-20.4 (17.4)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This primary analysis includes two 100 mg subjects who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0001
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing two subjects with no detectable simufilam in plasma, P=0.0001 and CFB was -23.7.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This primary analysis includes one 50 mg subject who showed no detectable simufilam in plasma at return visits (their exclusion was not prespecified in the Statistical Analysis Plan).
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0001
	Comments	P value adjusted for multiplicity of 6 co-primary endpoints (p<0.008 required for significance).
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.
Other Statistical Analysis		In a secondary analysis removing one subject with no detectable simufilam in plasma, P=0.0002 and CFB was -20.9.

7. Secondary Outcome

Title	Paired Associates Learning Test
Description	Cognitive test assessing episodic memory. Boxes are displayed on the screen and are "opened" in a randomized order. One or more of them contains a pattern. The patterns are then displayed in the middle of the screen, one at a time and the participant must select the box in which the pattern was originally located. If the participant makes an error, the boxes are opened in sequence again to remind the participant of the locations of the patterns. The number of boxes increases progressively to a total of 8.
Time Frame	Day 1 to Day 28

Outcome Measure Data

Analysis Population Description

The least impaired patients (11 or fewer errors, representing a ceiling effect) and patients with 54 or more errors (very poor performance suggesting not understanding the task) were removed from the analysis. Also removed were the 3 patients with no detectable drug in plasma, 2 patients with ≥25% non-compliance by pill counts, one patient with no baseline test and one who did not understand instructions per rater notes.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo oral tablets administered twice daily (BID) for 28 days.	Simufilam 100 mg oral tablets administered twice daily (BID) for 28 days.	Simufilam 50 mg oral tablets administered twice daily (BID) for 28 days.
Measure Participants	14	13	10
Mean (Standard Deviation) [Change from Day 1 in total errors]	-1.5 (8.5)	-4.5 (17.7)	-5.7 (13.6)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This study was not powered for statistical significance on cognitive measures.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Method of Estimation	Estimation Parameter	Effect size
	Estimated Value	0.23
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Reported effect size vs. placebo was calculated with Hedge's g (for groups of 20 or fewer) but was identical when calculated by Cohen's d.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This study was not powered for statistical significance on cognitive measures.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Method of Estimation	Estimation Parameter	Effect size
	Estimated Value	0.37
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Reported effect size vs. placebo was calculated with Hedge's g (for groups of 20 or fewer) but was identical when calculated by Cohen's d.

8. Secondary Outcome

Title	Spatial Working Memory Test
Description	Cognitive assessment of spatial working memory: A number of colored squares (boxes) are shown on the screen. By selecting the boxes and using a process of elimination, the subject should find one yellow 'token' in each of a number of boxes and use them to fill up an empty column on the right-hand side of the screen. The number of boxes is gradually increased to a total of 8 for the subjects to search. The colors and positions of the boxes are changed from trial to trial to discourage stereotyped search strategies.
Time Frame	Day 1 to Day 28

Outcome Measure Data

Analysis Population Description

Removed from analysis were the 3 patients with no detectable drug in plasma, 2 patients with ≥25% non-compliance by pill counts, one patient with no baseline test and one who did not understand instructions per rater notes.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo oral tablets administered twice daily (BID)	Simufilam, 100 mg oral tablets administered twice daily (BID)	Simufilam, 50 mg oral tablets administered twice daily (BID)
Measure Participants	22	18	17
Mean (Standard Deviation) [Change from Day 1 in total errors]	-0.41 (7.54)	-2.31 (7.45)	-3.35 (4.86)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This study was not powered for statistical significance on cognitive measures.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Method of Estimation	Estimation Parameter	Effect size
	Estimated Value	0.46
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Reported effect size vs. placebo was calculated with Hedge's g (for groups of 20 or fewer) but was almost identical (0.45) when calculated by Cohen's d.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This study was not powered for statistical significance on cognitive measures.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Method of Estimation	Estimation Parameter	Effect size
	Estimated Value	0.25
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments	Reported effect size vs. placebo was calculated with Hedge's g (for groups of 20 or fewer) but was identical when calculated by Cohen's d.

9. Secondary Outcome

Title	CSF IL-6, sTREM2, HMGB1, Albumin, IgG
Description	Change from Baseline (screening sample) to Day 28 in secondary CSF biomarkers of neuroinflammation and blood-brain barrier integrity
Time Frame	Screening to Day 28

Outcome Measure Data

Analysis Population Description

Three subjects who had no detectable simufilam in plasma at return visits were removed from analyses (two in the 100 mg arm and one in the 50 mg arm). As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. One additional subject in the 50 mg arm was missing a Day 28 CSF sample.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo oral tablets administered twice daily (BID)	Simufilam, 100 mg oral tablets administered twice daily (BID)	Simufilam, 50 mg oral tablets administered twice daily (BID)
Measure Participants	22	19	18
CSF IL-6	-1.1 (2.0)	-3.7 (1.8)	-3.3 (1.9)
CSF sTREM2	-77.3 (510)	-426 (274)	-424 (386)
CSF HMGB1	19.4 (172.3)	-143 (51.3)	-152 (50.1)
CSF albumin	-240 (1620)	-2292 (1760)	-1245 (1735)
CSF IgG	-574.8 (2518)	-2350 (2517)	-2444 (2097)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This analysis is for IL-6.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0078
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This analysis is for IL-6.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.019
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 3

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This analysis is for sTREM2.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0002
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 4

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This analysis is for sTREM2.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0007
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 5

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This analysis is for HMGB1.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0001
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 6

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This analysis is for HMGB1.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0001
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 7

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This analysis is for albumin.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.0001
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 8

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This analysis is for albumin.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.046
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 9

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	This analysis is for Immunoglobulin G.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.012
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

Statistical Analysis 10

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	This analysis is for Immunoglobulin G.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.014
	Comments	Secondary CSF biomarkers were not adjusted for multiplicity.
	Method	ANCOVA
	Comments	General Linear Model for the analysis of covariance (ANCOVA) with a two-sided 95% confidence interval and baseline measurement as the covariate.

10. Other Pre-specified Outcome

Title	Target Engagement Assays: Change From Baseline in Filamin A (FLNA) Linkages to alpha7 Nicotinic Acetylcholine Receptor (alpha7nAChR) and Toll-like Receptor 4 (TLR4) in Subject Lymphocytes
Description	FLNA linkages to these two receptors were assessed by densitometric quantitation of immunoblot bands of each receptor (detected by a specific antibody) in anti-FLNA precipitates. The measure is noted as a ratio to total FLNA.
Time Frame	Day 1 to Day 28

Outcome Measure Data

Analysis Population Description

Three subjects who had no detectable simufilam in plasma at any return visit were removed from analyses (two in the 100 mg arm and one in the 50 mg arm). As noted in the Participant Flow, one subject in the 50 mg arm did not complete the study. Two additional subjects in the 50 mg arm were missing Day 1 blood samples.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Placebo oral tablets administered twice daily (BID)	Simufilam, 100 mg oral tablets administered twice daily (BID)	Simufilam, 50 mg oral tablets administered twice daily (BID)
Measure Participants	22	19	17
FLNA - alpha7nAChR linkage	-0.07 (0.19)	-0.24 (0.16)	-0.23 (0.13)
FLNA - TLR4 linkage	-0.05 (0.18)	-0.19 (0.14)	-0.19 (0.11)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	Change from baseline in FLNA linkage to alpha7nAChR in subject lymphocytes. FLNA linkage to alpha7nAChR was expressed as the ratio of densitometric units of immunoblot bands of alpha7nAChR (probed with a specific antibody) to densitometric units of total FLNA.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.005
	Comments	As secondary endpoints, lymphocyte biomarkers were not adjusted for multiplicity.
	Method	ANOVA
	Comments	ANOVA was used instead of ANCOVA due to the range in baseline values; it was deemed more appropriate to compare to each subject's own baseline value.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	Change from baseline in FLNA linkage to alpha7nAChR in subject lymphocytes. FLNA linkage to alpha7nAChR was expressed as ratios of densitometric units of immunoblot bands of alpha7nAChR (probed with a specific antibody) to densitometric units of total FLNA.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.009
	Comments	As secondary endpoints, lymphocyte biomarkers were not adjusted for multiplicity.
	Method	ANOVA
	Comments	ANOVA was used instead of ANCOVA due to the range in baseline values; it was deemed more appropriate to compare to each subject's own baseline value.

Statistical Analysis 3

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments	Change from baseline in FLNA linkage to TLR4 in subject lymphocytes. FLNA linkage to TLR4 was expressed as the ratio of densitometric units of immunoblot bands of TLR4 (probed with a specific antibody) to densitometric units of total FLNA.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.01
	Comments	As secondary endpoints, lymphocyte biomarkers were not adjusted for multiplicity.
	Method	ANOVA
	Comments	ANOVA was used instead of ANCOVA due to the range in baseline values; it was deemed more appropriate to compare to each subject's own baseline value.

Statistical Analysis 4

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments	Change from baseline in FLNA linkage to TLR4 in subject lymphocytes. FLNA linkage to TLR4 was expressed as the ratio of densitometric units of immunoblot bands of TLR4 (probed with a specific antibody) to densitometric units of total FLNA.
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.01
	Comments	As secondary endpoints, lymphocyte biomarkers were not adjusted for multiplicity.
	Method	ANOVA
	Comments	ANOVA was used instead of ANCOVA due to the range in baseline values; it was deemed more appropriate to compare to each subject's own baseline value.

11. Other Pre-specified Outcome

Title	Plasma P-tau181
Description	Percent change in plasma P-tau181
Time Frame	Day 1 to Day 28

Outcome Measure Data

Analysis Population Description

For plasma p-tau181 only: 4 subjects (1 in placebo, 2 in 50 mg and 1 in 100 mg arms) were removed because Coefficients of Variation (CVs) between duplicate measurements for either Day 1 or Day 28 samples were >15% on repeat assay (both Day 1 & Day 28 for a subject were repeated if CVs of either Day 1 or Day 28 were >11%). Two outliers were removed (1 in placebo [1.2 to 4.8 pg/ml] and 1 in 100 mg [2.1 to 5.1 pg/ml] arms) for increases >150% & > 2.5 pg/mL. Missing Day 1 blood samples: 2 in 50 mg.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125) 100 mg Tablets Cohort	Simufilam (PTI-125) 50 mg Tablets Cohort
Arm/Group Description	Subjects administered placebo oral tablets twice daily (BID) Placebo oral tablet: Oral placebo tablet	Subjects administered simufilam (PTI-125) 100 mg oral tablets twice daily (BID) Simufilam 50 mg oral tablet: Simufilam 50 mg oral tablet	Subjects administered simufilam (PTI-125) 50 mg oral tablets twice daily (BID) Simufilam 100 mg tablet: Simufilam 100 mg oral tablet
Measure Participants	20	17	15
Mean (Standard Deviation) [Percent change]	20.7 (49)	-16.5 (29)	-15.1 (36)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.01
	Comments
	Method	ANOVA
	Comments	ANOVA followed by Dunnett's multiple comparisons test.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.02
	Comments
	Method	ANOVA
	Comments	ANOVA followed by Dunnett's multiple comparisons test.

Statistical Analysis 3

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.009
	Comments
	Method	ANOVA
	Comments	This p value is for the main effect of treatment of the ANOVA.

12. Other Pre-specified Outcome

Title	Percent Change From Baseline in SavaDx, a Novel Plasma Biomarker
Description	SavaDx is a novel plasma biomarker
Time Frame	Day 1 to Day 28

Outcome Measure Data

Analysis Population Description

One subject in the 50 mg arm and two subjects in the 100 mg arm were omitted because these subjects showed no detectable plasma simufilam at any return visit. Two additional subjects in the 50 mg group were missing baseline plasma samples.

Arm/Group Title	Placebo Cohort	Simufilam (PTI-125), 100 mg Tablets Cohort	Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Subjects administered placebo oral tablets twice daily (BID)	Subjects administered simufilam 100 mg oral tablets twice daily (BID)	Subjects administered simufilam 50 mg oral tablets twice daily (BID)
Measure Participants	22	19	17
Mean (Standard Deviation) [Percent change]	-3.2 (62)	-47.8 (19)	-44.1 (35)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 100 mg Tablets Cohort
	Comments
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.003
	Comments
	Method	ANOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Placebo Cohort, Simufilam (PTI-125), 50 mg Tablets Cohort
	Comments
	Type of Statistical Test	Superiority
	Comments
Statistical Test of Hypothesis	p-Value	0.016
	Comments
	Method	ANOVA
	Comments

Adverse Events

Time Frame	28 days
Arm/Group Title	Placebo Cohort		Simufilam (PTI-125), 100 mg Tablets Cohort		Simufilam (PTI-125), 50 mg Tablets Cohort
Arm/Group Description	Subjects administered placebo tablets twice daily (BID) for 28 days.		Subjects administered simufilam 100 mg tablets twice daily (BID) for 28 days.		Subjects administered simufilam 50 mg tablets twice daily (BID) for 28 days.
All Cause Mortality
	Placebo Cohort		Simufilam (PTI-125), 100 mg Tablets Cohort		Simufilam (PTI-125), 50 mg Tablets Cohort
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/22 (0%)		0/21 (0%)		0/21 (0%)
Serious Adverse Events
	Placebo Cohort		Simufilam (PTI-125), 100 mg Tablets Cohort		Simufilam (PTI-125), 50 mg Tablets Cohort
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/22 (0%)		0/21 (0%)		0/21 (0%)
Other (Not Including Serious) Adverse Events
	Placebo Cohort		Simufilam (PTI-125), 100 mg Tablets Cohort		Simufilam (PTI-125), 50 mg Tablets Cohort
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	12/22 (54.5%)		9/21 (42.9%)		4/21 (19%)
Gastrointestinal disorders
Nausea	2/22 (9.1%)	2	1/21 (4.8%)	1	0/21 (0%)	0
General disorders
Any adverse event	12/22 (54.5%)	20	9/21 (42.9%)	15	4/21 (19%)	9
Metabolism and nutrition disorders
Fatigue	2/22 (9.1%)	2	0/21 (0%)	0	1/21 (4.8%)	1
Nervous system disorders
Headache	3/22 (13.6%)	3	2/21 (9.5%)	2	1/21 (4.8%)	1

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

The only disclosure restriction on the PI is that the sponsor can review results communications prior to public release and can embargo communications regarding trial results for a period that is less than or equal to 60 days. The sponsor cannot require changes to the communication and cannot extend the embargo.