Effect of Cilostazol on Endothelial Progenitor Cells and Endothelial Function in Coronary Artery Disease

Phase 4

• Conditions: Coronary Artery Disease
• Interventions: Drug: Dummy Placebo; Drug: Cilostazol

• Registration Number: NCT02174939
• Lead Sponsor: National Cheng-Kung University Hospital

Brief Summary

1. The number and function of circulating endothelial progenitor cells (EPCs) are inversely associated with coronary risk factors and atherosclerotic diseases such as coronary artery disease (CAD) and cardiovascular high risk.

2. This double-blind, randomized, placebo-controlled trial to evaluate the effects of cilostazol on human early EPCs and endothelial function as well as the potential mechanisms of action in patients with CAD and cardiovascular high risk.

Detailed Description

1. titration of drugs

1. run-in period: eligible subjects are screened and baseline blood samples are obtained

2. study period: 12 weeks

* subjects with cilostazol and subjects with dummy placebo

* On the first day after the end of the study period, the follow-up data are obtained by the same procedure

3. blood sampling and measurement of serum biomarkers

* obtained from peripheral veins in all study subjects at the run-in period and the end of the treatment period of the study

* sent for isolation, cell culture, and assays of human EPCs

* also stored for enzyme-linked immunosorbent assay (Stromal cell derived factor-alfa1, adiponectin, soluble thrombomodulin, vascular endothelial growth factor)

2. assays of human EPCs

1. colony formation by EPCs

2. quantification of EPCs and apoptotic endothelial cells

3. chemotactic motility, proliferation/viability and apoptosis assays

3. measurement of flow-mediated dilatation (FMD) of left brachial artery by sonography

4. assessment of long-term cardiovascular outcomes

Study Timeline

• Study Start: 2014-02
• Study First Submitted: 2014-06-22
• Study First Submitted QC: 2014-06-24
• Study First Posted: 2014-06-26
• Status Verified: 2015-10
• Last Update Submitted: 2015-10-11
• Last Update Posted: 2015-10-14
• Primary Completion: 2017-10
• Study Completion: 2017-12

Recruitment & Eligibility

• Status: UNKNOWN
• Sex: All
• Target Recruitment: 300

Inclusion Criteria

• stable CAD documented by stress test, computed tomography angiography or coronary angiography or
• old myocardial infarction (>6 months)
• history and evidence of CAD
• history and evidence of cerebrovascular accident
• history and evidence of peripheral artery disease
• diabetes mellitus
• metabolic syndrome
• stage 3 to 5 chronic kidney disease
• at least 2 of the followings: male ≥45 years old or female ≥55 years old; hypertension; current or past 3-year tobacco smoking; hyperlipidemia; family history of premature CAD (male <55 years old or female <65 years old)

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Exclusion Criteria

• unstable CAD
• have plan to do percutaneous intervention or bypass surgery for CAD or peripheral artery disease within recent 3 months
• severe liver dysfunction (transaminases >10 times of upper normal limit, history of liver cirrhosis, or hepatoma)
• left ventricular ejection fraction (<50% by echocardiography)
• documented active malignancy
• chronic inflammatory disease
• known drug allergy history for cilostazol
• current use of cilostazol or any other cAMP-elevator
• premenopausal women

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Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Arm && Interventions

Group	Intervention	Description
Dummy Placebo	Dummy Placebo	One tablet twice per day for 12 weeks
Cilostazol	Cilostazol	One tablet (100 mg) twice per day for 12 weeks

Primary Outcome Measures

Name	Time	Method
Circulating EPCs Number	3 months	Peripheral blood mononuclear cells (one million cells in each) are suspended in 100 µL phosphate-buffered saline and incubated for 30 min with monoclonal antibodies against human peridinin chlorophyll protein-conjugated cluster of differentiation antigen-45, phycoerythrin-conjugated anti-human cluster of differentiation antigen-34 antibody and anti-human kinase insert domain receptor (KDR) antibody conjugated with Alexa Flour 647. Cells are washed and analyzed on a FACSCalibur flow cytometer with 100,000 events in the lymphocyte gate. EPCs, which are defined as negative for cluster of differentiation antigen-45 and positive for cluster of differentiation antigen-34 and KDR. Based on the peripheral blood mononuclear cell counts, the absolute number of circulating EPCs/µL is calculated.

Secondary Outcome Measures

Name	Time	Method
Composite Major Adverse Cardiovascular Events (MACE)	at least 1 year	This composite endpoint includes cardiovascular death, nonfatal myocardial infarction, nonfatal stroke, congestive heart failure hospitalization, and target vessel revascularization.
Viability (Proliferation) of EPCs	3 months	250,000 cells are seeded in each well of a 96-well plate and the cells are added with 200 μl of the culture medium and incubated at 37°C. Medium change is performed 3 days later. On 7th day, the plate is then re-incubated with 100 μl fresh medium and additional 50 μl of 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2 Hydrogen-Tetrazolium-5-Carboxanilide reagent, and further incubated in dark at 37°C for 4 h. After incubation, the orange colored complex formed is read at 450 nm using a microplate reader with a 450 nm reference filter.
composite major coronary events	at least 1 year	This composite endpoint includes fatal or nonfatal myocardial infarction, recurrent angina pectoris, and target vessel revascularization.

Trial Locations

Locations (1)

National Cheng Kung University Hospital; 🇨🇳 Tainan, Taiwan