Effects of Methylene Blue in Healthy Aging, Mild Cognitive Impairment and Alzheimer's Disease (MB2)

• ClinicalTrials.gov Identifier: NCT02380573
• Recruitment Status: Suspended
• First Posted: March 5, 2015
• Last Update Posted: May 11, 2020
• Sponsor: The University of Texas Health Science Center at San Antonio
• Collaborator: Texas Alzheimer's Research and Care Consortium (TARCC)
• Information provided by (Responsible Party): The University of Texas Health Science Center at San Antonio

Tracking Information

• First Submitted Date: February 24, 2015
• First Posted Date: March 5, 2015
• Last Update Posted Date: May 11, 2020
• Study Start Date: July 2015
• Estimated Primary Completion Date: July 2022 (Final data collection date for primary outcome measure)

Current Primary Outcome Measures (submitted: May 6, 2019)

• Working memory task [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  fMRI measurement of task blocked activation
• Working memory task response [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  Working memory task behavioral measures (ie. correct number of responses)
• Episodic memory task [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  fMRI measurement of task blocked activation
• Episodic memory task response [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  Face-Name Task behavioral measures (ie. correct recalls)
• Sustained attention task [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  fMRI measurement of task blocked activation
• Sustained attention task reaction time [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  Psychomotor vigilance task (PVT) behavioral measures (ie. reaction time)
• Neuropsychological battery composite score [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  TARCC designed neurocognitive tests

Original Primary Outcome Measures (submitted: February 27, 2015)

• Working memory task fMRI [ Time Frame: baseline, 2 weeks ± 3 days ]
  fMRI task blocked activation
• Working memory task response [ Time Frame: baseline, 2 weeks ± 3 days ]
  Working memory task behavioral measures (ie. correct number of responses)
• Episodic memory task fMRI [ Time Frame: baseline, 2 weeks ± 3 days ]
  fMRI task blocked activation
• Episodic memory task response [ Time Frame: baseline, 2 weeks ± 3 days ]
  Face-Name Task behavioral measures (ie. correct recalls)
• Sustained attention task fMRI [ Time Frame: baseline, 2 weeks ± 3 days ]
  fMRI task blocked activation
• Sustained attention task reaction time [ Time Frame: baseline, 2 weeks ± 3 days ]
  Psychomotor vigilance task (PVT) behavioral measures (ie. reaction time)
• Neuropsychological battery composite score [ Time Frame: baseline, 2 weeks ± 3 days ]
  TARCC designed neurocognitive tests

Current Secondary Outcome Measures (submitted: May 6, 2019)

Cerebral blood flow measures [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]

Resting measurements will be used to assess response and CBF using fMRI

Original Secondary Outcome Measures (submitted: February 27, 2015)

Cerebral blood flow fMRI measures [ Time Frame: baseline, 2 weeks ± 3 days ]

Resting measurements will be used to assess response and CBF

Current Other Pre-specified Outcome Measures (submitted: May 6, 2019)

• Functional Connectivity measures [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  fMRI measurements will be obtained while the subject rests in the scanner
• CO2 challenge [ Time Frame: baseline, change from baseline at 2 weeks ± 3 days, change from baseline at 12 weeks ± 3 days ]
  Cerebral blood flow measurements will be acquired during a brief (3-5 minutes) inhalation of medical-grade 5% CO2 in air.

Original Other Pre-specified Outcome Measures (submitted: February 27, 2015)

• Functional Connectivity fMRI measures [ Time Frame: baseline, 2 weeks ± 3 days ]
  fMRI measurements will be obtained while the subject rests in the scanner
• CO2 challenge [ Time Frame: Baseline ]
  Cerebral blood flow measurements will be acquired during a brief (3-5 minutes) inhalation of medical-grade 5% CO2 in air only at baseline for fMRI signal calibration.

Descriptive Information

• Brief Title: Effects of Methylene Blue in Healthy Aging, Mild Cognitive Impairment and Alzheimer's Disease
• Official Title: Cognitive and Functional Connectivity Effects of Methylene Blue in Healthy Aging, Mild Cognitive Impairment and Alzheimer's Disease
• Brief Summary: A double-blind, placebo-controlled study that aims to investigate the effect of 2-week and 12-week administration of USP methylene blue (MB) on cerebral blood flow, functional connectivity, memory and attention cognitive abilities using fMRI and behavioral measures in healthy aging, mild cognitive impairment (MCI) and mild Alzheimer's disease (AD) subjects.

Detailed Description

Healthy aging and aging human subjects with mild cognitive impairment and mild Alzheimer's disease from the TARCC cohort and South Texas will be studied using a double-blind, placebo-controlled design. After informed consent and familiarity with the tasks and the MRI environment, the subject will enter an MRI scanner and perform the following 6 tasks.

fMRI and behavioral data will be collected simultaneously while inside the scanner.

Delayed match-to-sample task: The subject views a pattern for a few seconds and then is prompted to recall the memorized pattern using a response system (approx. 10 mins).

Face-name task: The subject is shown blocks of stimuli where a novel or familiar face is paired with a name. In a later run, the subjects are asked whether the correct name is matched with the correct face. (approx. 10 mins).

Psychomotor vigilance task: The subject receives a visual cue that alerts them to press a button as fast as possible. (approx. 10 mins).

Cerebral Blood Flow and Resting State fMRI: Subject scanned with eyes closed and told to not think about a particular topic, each lasting about 10 minutes.

fMRI data acquisition: fMRI and neuropsychological battery measurements will be made before the intervention. These measurements will then be repeated after 2 weeks and 12 weeks.

fMRI will image changes in regional brain activity associated with these tasks. The MRI pulse sequences include diffusion tensor imaging, standard and non-invasive anatomical and quantitative MRI for coregistration and blood-oxygen-level dependent (BOLD) fMRI.

CO2 challenge: Cerebral blood flow measurements will be obtained while the subject rests in the scanner after administration of medical-grade 5% CO2 in air for 3-5 minutes. This will be repeated on weeks 2 and 12.

Data analysis: Standard fMRI analysis will be analyzed using established fMRI software. Statistical parametric analysis will be performed to generate activation maps. fMRI data will be corrected for multiple comparisons using a false discovery rate (q < 0.05) and threshold for cluster values to conservatively control for type I error. Behavioral data will be analyzed with paired t-test and ANOVA calculations used for group comparison with p < 0.05 (with Bonferroni correction) considered statistically significant.

Expected results: The investigators predict that, compared to placebo, MB will: i) improve working memory retention in a delayed match-to-sample task by memory performance and enhanced fMRI responses in the prefrontal cortex and parietal lobes, ii) improve episodic memory as determined by fMRI activation in the hippocampus, medial temporal lobes and prefrontal cortex iii) reduce reaction time in a psychomotor vigilance test and enhance fMRI responses within a cortical sustained attention network iv) improve CBF and v) improve fMRI connectivity in default mode and visuospatial and memory networks/subnetworks. The fMRI and behavioral performance effects on memory will be greater in the MCI and mild AD groups than in the healthy aging group. The effects will be greater in the MCI and AD groups than in the control groups.

Power analysis: Sample sizes were calculated using an fMRI power tool based on pilot data from the current study for a power of 80%, alpha = 0.05, False Discovery Rate < 0.05, to detect statistical difference between MB and placebo23. The investigators estimate they will need 20-25 subjects per arm of group (complete studies) and thus will recruit 200-240 subjects to account for potential failed studies.

• Study Type: Interventional
• Study Phase: Phase 2
• Study Design: Allocation: Randomized; Intervention Model: Single Group Assignment; Masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor); Primary Purpose: Treatment

Condition

• Mild Cognitive Impairment
• MCI
• Aging
• Alzheimer's Disease
• AD

Intervention

• Drug: Methylene Blue
  Other Name: Phenothiazin-5-ium, 3, 7-bis (dimethylamino)-chloride, trihydrate
• Drug: FD&C Blue # 2
  Other Name: Placebo
• Drug: Phenazopyridine hydrochloride
  Other Name: Azo

Study Arms

• Experimental: Healthy Aging MB
  Methylene Blue (USP grade, 282mg oral, daily, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: Methylene Blue
  • Drug: Phenazopyridine hydrochloride
• Placebo Comparator: Healthy Aging Placebo
  Drug: FD&C Blue # 2 (USP grade, 282 mg oral, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: FD&C Blue # 2
  • Drug: Phenazopyridine hydrochloride
• Experimental: Mild Cognitive Impairment (MCI) MB
  Methylene Blue (USP grade, 282 mg oral, daily, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: Methylene Blue
  • Drug: Phenazopyridine hydrochloride
• Placebo Comparator: Mild Cognitive Impairment (MCI) Placebo
  Drug: FD&C Blue # 2 (USP grade, 282 mg oral, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: FD&C Blue # 2
  • Drug: Phenazopyridine hydrochloride
• Experimental: Mild Alzheimer's Disease (AD) MB
  Methylene Blue (USP grade, 282 mg oral, daily, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: Methylene Blue
  • Drug: Phenazopyridine hydrochloride
• Placebo Comparator: Mild Alzheimer's Disease (AD) Placebo
  Drug: FD&C Blue # 2 (USP grade, 282 mg oral, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: FD&C Blue # 2
  • Drug: Phenazopyridine hydrochloride
• Experimental: Healthy Middle Age MB
  Methylene Blue (USP grade, 282 mg oral, daily, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: Methylene Blue
  • Drug: Phenazopyridine hydrochloride
• Placebo Comparator: Healthy Middle Age Placebo
  Drug: FD&C Blue # 2 (USP grade, 282 mg oral, 2 weeks, 12 weeks) Phenazopyridine hydrochloride (97.5 mg oral, 2 weeks, 12 weeks)
  Interventions:

  • Drug: FD&C Blue # 2
  • Drug: Phenazopyridine hydrochloride

Publications *

• Bruchey AK, Gonzalez-Lima F. Behavioral, Physiological and Biochemical Hormetic Responses to the Autoxidizable Dye Methylene Blue. Am J Pharmacol Toxicol. 2008 Jan 1;3(1):72-79.
• Rojas JC, Bruchey AK, Gonzalez-Lima F. Neurometabolic mechanisms for memory enhancement and neuroprotection of methylene blue. Prog Neurobiol. 2012 Jan;96(1):32-45. doi: 10.1016/j.pneurobio.2011.10.007. Epub 2011 Nov 3. Review.
• Telch MJ, Bruchey AK, Rosenfield D, Cobb AR, Smits J, Pahl S, Gonzalez-Lima F. Effects of post-session administration of methylene blue on fear extinction and contextual memory in adults with claustrophobia. Am J Psychiatry. 2014 Oct;171(10):1091-8. doi: 10.1176/appi.ajp.2014.13101407.
• Lin AL, Poteet E, Du F, Gourav RC, Liu R, Wen Y, Bresnen A, Huang S, Fox PT, Yang SH, Duong TQ. Methylene blue as a cerebral metabolic and hemodynamic enhancer. PLoS One. 2012;7(10):e46585. doi: 10.1371/journal.pone.0046585. Epub 2012 Oct 9.
• Huang S, Du F, Shih YY, Shen Q, Gonzalez-Lima F, Duong TQ. Methylene blue potentiates stimulus-evoked fMRI responses and cerebral oxygen consumption during normoxia and hypoxia. Neuroimage. 2013 May 15;72:237-42. doi: 10.1016/j.neuroimage.2013.01.027. Epub 2013 Jan 26.
• Talley Watts L, Long JA, Chemello J, Van Koughnet S, Fernandez A, Huang S, Shen Q, Duong TQ. Methylene blue is neuroprotective against mild traumatic brain injury. J Neurotrauma. 2014 Jun 1;31(11):1063-71. doi: 10.1089/neu.2013.3193. Epub 2014 Apr 8.
• Long JA, Watts LT, Chemello J, Huang S, Shen Q, Duong TQ. Multiparametric and longitudinal MRI characterization of mild traumatic brain injury in rats. J Neurotrauma. 2015 Apr 15;32(8):598-607. doi: 10.1089/neu.2014.3563. Epub 2015 Jan 22.
• Rodriguez P, Jiang Z, Huang S, Shen Q, Duong TQ. Methylene blue treatment delays progression of perfusion-diffusion mismatch to infarct in permanent ischemic stroke. Brain Res. 2014 Nov 7;1588:144-9. doi: 10.1016/j.brainres.2014.09.007. Epub 2014 Sep 8.
• Shen Q, Du F, Huang S, Rodriguez P, Watts LT, Duong TQ. Neuroprotective efficacy of methylene blue in ischemic stroke: an MRI study. PLoS One. 2013 Nov 21;8(11):e79833. doi: 10.1371/journal.pone.0079833. eCollection 2013.
• Peter C, Hongwan D, Küpfer A, Lauterburg BH. Pharmacokinetics and organ distribution of intravenous and oral methylene blue. Eur J Clin Pharmacol. 2000 Jun;56(3):247-50.
• Gonzalez-Lima F, Bruchey AK. Extinction memory improvement by the metabolic enhancer methylene blue. Learn Mem. 2004 Sep-Oct;11(5):633-40.
• Naylor GJ, Martin B, Hopwood SE, Watson Y. A two-year double-blind crossover trial of the prophylactic effect of methylene blue in manic-depressive psychosis. Biol Psychiatry. 1986 Aug;21(10):915-20.
• Mackworth JF. Vigilance, arousal, and habituation. Psychol Rev. 1968 Jul;75(4):308-22. Review.
• Rombouts SA, Barkhof F, Goekoop R, Stam CJ, Scheltens P. Altered resting state networks in mild cognitive impairment and mild Alzheimer's disease: an fMRI study. Hum Brain Mapp. 2005 Dec;26(4):231-9.
• Wang L, Li H, Liang Y, Zhang J, Li X, Shu N, Wang YY, Zhang Z. Amnestic mild cognitive impairment: topological reorganization of the default-mode network. Radiology. 2013 Aug;268(2):501-14. doi: 10.1148/radiol.13121573. Epub 2013 Mar 12.

Recruitment Information

• Recruitment Status: Suspended
• Actual Enrollment (submitted: May 4, 2019): 117
• Original Estimated Enrollment (submitted: February 27, 2015): 115
• Estimated Study Completion Date: July 2023
• Estimated Primary Completion Date: July 2022 (Final data collection date for primary outcome measure)

Eligibility Criteria

Inclusion Criteria for all subjects:

1. 45-89 years old
2. All genders
3. All minorities
4. English, Spanish, or multilingual speakers
5. Postmenopausal or surgically sterile females only.
6. Inclusion for MCI group only: participants will meet the criteria for amnestic and non-amnestic MCI such as those currently used by Texas Alzheimer's Research and Care Consortium (TARCC) consensus diagnosis
7. Inclusion for AD group only: Alzheimer's Early-stage, sporadic-type

Exclusion Criteria:

1. Pregnancy or breastfeeding
2. Contraindication for MRI (Claustrophobia and magnetic metal implants)
3. Glucose-6-phosphate deficiency, methemoglobinemia
4. Allergy to MB
5. Color-blindness
6. Craniotomy, craniectomy or endovascular neurosurgery
7. A current diagnosis of stroke, transient ischemic attack (TIA), any primary neurodegenerative disorder, or any other causes of neuropsychologic disturbances or secondary dementia (MCI or AD does not exclude subject)
8. A serious intercurrent illness likely to cause death within the next 5 years, such as terminal cancer
9. Alcohol and/or drug abuse
10. Any detection of an unknown disease process (eg. new tumor) on the study's neuroimaging at the discretion of the investigators
11. A systolic blood pressure ≥180 mmHg and/or a diastolic blood pressure ≥105 mmHg
12. Severe difficulty or an inability to perform any one of the 6 Katz Activities of Daily Living
13. Patients who are unlikely to comply with trial visit schedule or with trial medication,
14. On any psychiatric serotonergic antidepressant medication or psychotropic medication within the last 5 weeks
15. Diagnosis of epilepsy, traumatic brain injury with loss of consciousness, psychosis, panic attacks,
16. Chronic kidney disease, cirrhosis, liver or renal transplants
17. Known hypersensitivity to thiazide diuretics and phenothiazines
18. Any other condition, which in the opinion of the investigator, would put the participant at risk and warrant exclusion from the study

Sex/Gender

• Sexes Eligible for Study: All

• Ages: 45 Years to 89 Years (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes
• Contacts: Contact information is only displayed when the study is recruiting subjects
• Listed Location Countries: United States

Administrative Information

• NCT Number: NCT02380573
• Other Study ID Numbers: HSC20150410H
• Has Data Monitoring Committee: Yes
• U.S. FDA-regulated Product: Not Provided
• IPD Sharing Statement: Not Provided
• Responsible Party: The University of Texas Health Science Center at San Antonio
• Study Sponsor: The University of Texas Health Science Center at San Antonio
• Collaborators: Texas Alzheimer's Research and Care Consortium (TARCC)
• Investigators: Not Provided
• PRS Account: The University of Texas Health Science Center at San Antonio
• Verification Date: May 2020