<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT02194686</url>
  </required_header>
  <id_info>
    <org_study_id>B-BR-101-121</org_study_id>
    <secondary_id>NCKUH-10203022</secondary_id>
    <nct_id>NCT02194686</nct_id>
  </id_info>
  <brief_title>Effect of Cilostazol on Endothelial Progenitor Cells and Endothelial Function in High Risk for Cardiovascular Disease</brief_title>
  <official_title>Cilostazol Enhances the Number and Functions of Circulating Endothelial Progenitor Cells Mediated Through Multiple Mechanisms in Patients With High Risk for Cardiovascular Disease</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>National Cheng-Kung University Hospital</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Department of Health, Executive Yuan, R.O.C. (Taiwan)</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>National Cheng-Kung University Hospital</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      1. The number and function of circulating endothelial progenitor cells (EPCs) are inversely&#xD;
           associated with coronary risk factors and atherosclerotic diseases.&#xD;
&#xD;
        2. This double-blind, randomized, placebo-controlled trial to evaluate the effects of&#xD;
           cilostazol on human early EPCs and endothelial function as well as the potential&#xD;
           mechanisms of action in patients with high risk for cardiovascular disease.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      1. titration of drugs&#xD;
&#xD;
             1. run-in period: eligible subjects are screened and baseline blood samples are&#xD;
                obtained&#xD;
&#xD;
             2. study period: 12 weeks&#xD;
&#xD;
                  -  subjects with cilostazol and subjects with dummy placebo&#xD;
&#xD;
                  -  On the first day after the end of the study period, the follow-up data are&#xD;
                     obtained by the same procedure&#xD;
&#xD;
             3. blood sampling and measurement of serum biomarkers&#xD;
&#xD;
                  -  obtained from peripheral veins in all study subjects at the run-in period and&#xD;
                     the end of the treatment period of the study&#xD;
&#xD;
                  -  sent for isolation, cell culture, and assays of human EPCs&#xD;
&#xD;
                  -  also stored for enzyme-linked immunosorbent assay (stromal cell derived&#xD;
                     factor-alfa1, adiponectin, soluble thrombomodulin, vascular endothelial growth&#xD;
                     factor)&#xD;
&#xD;
        2. assays of human EPCs&#xD;
&#xD;
             1. colony formation by EPCs&#xD;
&#xD;
             2. quantification of EPCs and apoptotic endothelial cells&#xD;
&#xD;
             3. chemotactic motility, proliferation/viability and apoptosis assays&#xD;
&#xD;
        3. measurement of flow-mediated dilatation (FMD) of left brachial artery by sonography&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date>January 2013</start_date>
  <completion_date type="Actual">August 2014</completion_date>
  <primary_completion_date type="Actual">March 2014</primary_completion_date>
  <phase>Phase 4</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Randomized</allocation>
    <intervention_model>Parallel Assignment</intervention_model>
    <primary_purpose>Treatment</primary_purpose>
    <masking>Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Circulating EPCs Number</measure>
    <time_frame>3 months</time_frame>
    <description>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.</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Viability (Proliferation) of EPCs</measure>
    <time_frame>3 months</time_frame>
    <description>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.</description>
  </secondary_outcome>
  <other_outcome>
    <measure>FMD of the Brachial Artery</measure>
    <time_frame>3 months</time_frame>
    <description>FMD in response to reactive hyperemia is measured in the left brachial artery in a quiet, temperature-controlled room after 10 min of bed rest. A high-resolution ultrasound machine equipped with a 7.5 mega Hertz linear array probe is used for the study. Arterial diameters are measured at the baseline and during reactive hyperemia. FMD is calculated as the percentage change in diameter compared with the baseline.</description>
  </other_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">71</enrollment>
  <condition>Cardiovascular Diseases</condition>
  <arm_group>
    <arm_group_label>Cilostazol</arm_group_label>
    <arm_group_type>Active Comparator</arm_group_type>
    <description>One tablet (100 mg) twice per day for 12 weeks</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Dummy Placebo</arm_group_label>
    <arm_group_type>Placebo Comparator</arm_group_type>
    <description>One tablet twice per day for 12 weeks</description>
  </arm_group>
  <intervention>
    <intervention_type>Drug</intervention_type>
    <intervention_name>Cilostazol</intervention_name>
    <description>One tablet (100 mg) twice per day for 12 weeks</description>
    <arm_group_label>Cilostazol</arm_group_label>
    <other_name>Pletaal (brand name)</other_name>
  </intervention>
  <intervention>
    <intervention_type>Drug</intervention_type>
    <intervention_name>Dummy Placebo</intervention_name>
    <description>One tablet twice per day for 12 weeks</description>
    <arm_group_label>Dummy Placebo</arm_group_label>
    <other_name>Placebo</other_name>
    <other_name>Control</other_name>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria: high-risk patients who have at least one of the following situations&#xD;
        without pre-existing cardiovascular disease including peripheral artery disease or coronary&#xD;
        artery disease:&#xD;
&#xD;
          -  type 2 diabetes mellitus&#xD;
&#xD;
          -  metabolic syndrome&#xD;
&#xD;
          -  stage 3 (or more advanced) chronic kidney disease&#xD;
&#xD;
          -  2 or more coronary risk factors (male &gt; 45 years or female &gt; 55 years, hypertension,&#xD;
             tobacco smoking, hyperlipidemia, family history of cardiovascular disease)&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  ankle-brachial index less than 0.9 or more than 1.3 in one or both legs&#xD;
&#xD;
          -  significant stenosis (more than 50% as compared to reference vessel) in peripheral&#xD;
             artery on image study&#xD;
&#xD;
          -  symptoms suggesting peripheral artery disease in at least one leg&#xD;
&#xD;
          -  clinical or electrocardiographic evidence of coronary artery disease&#xD;
&#xD;
          -  clinical evidence of cerebrovascular disease&#xD;
&#xD;
          -  severe liver dysfunction (transaminases &gt;10 times of upper normal limit, history of&#xD;
             liver cirrhosis, or hepatoma)&#xD;
&#xD;
          -  left ventricular ejection fraction (&lt;50% by echocardiography)&#xD;
&#xD;
          -  documented active malignancy&#xD;
&#xD;
          -  chronic inflammatory disease&#xD;
&#xD;
          -  known drug allergy history for cilostazol&#xD;
&#xD;
          -  current use of cilostazol or any other cAMP-elevator&#xD;
&#xD;
          -  premenopausal women&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>20 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Ting-Hsing Chao, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>National Cheng-Kung University Hospital</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>National Cheng Kung University Hospital</name>
      <address>
        <city>Tainan</city>
        <zip>704</zip>
        <country>Taiwan</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Taiwan</country>
  </location_countries>
  <reference>
    <citation>Chao TH, Tseng SY, Li YH, Liu PY, Cho CL, Shi GY, Wu HL, Chen JH. A novel vasculo-angiogenic effect of cilostazol mediated by cross-talk between multiple signalling pathways including the ERK/p38 MAPK signalling transduction cascade. Clin Sci (Lond). 2012 Aug 1;123(3):147-59. doi: 10.1042/CS20110432.</citation>
    <PMID>22339730</PMID>
  </reference>
  <reference>
    <citation>Biscetti F, Pecorini G, Straface G, Arena V, Stigliano E, Rutella S, Locatelli F, Angelini F, Ghirlanda G, Flex A. Cilostazol promotes angiogenesis after peripheral ischemia through a VEGF-dependent mechanism. Int J Cardiol. 2013 Aug 10;167(3):910-6. doi: 10.1016/j.ijcard.2012.03.103. Epub 2012 Apr 2.</citation>
    <PMID>22473072</PMID>
  </reference>
  <reference>
    <citation>Chao TH, Tseng SY, Chen IC, Tsai YS, Huang YY, Liu PY, Ou HY, Li YH, Wu HL, Cho CL, Tsai LM, Chen JH. Cilostazol enhances mobilization and proliferation of endothelial progenitor cells and collateral formation by modifying vasculo-angiogenic biomarkers in peripheral arterial disease. Int J Cardiol. 2014 Mar 15;172(2):e371-4. doi: 10.1016/j.ijcard.2013.12.295. Epub 2014 Jan 11. No abstract available.</citation>
    <PMID>24439864</PMID>
  </reference>
  <verification_date>October 2015</verification_date>
  <study_first_submitted>July 16, 2014</study_first_submitted>
  <study_first_submitted_qc>July 16, 2014</study_first_submitted_qc>
  <study_first_posted type="Estimate">July 18, 2014</study_first_posted>
  <last_update_submitted>October 11, 2015</last_update_submitted>
  <last_update_submitted_qc>October 11, 2015</last_update_submitted_qc>
  <last_update_posted type="Estimate">October 14, 2015</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>cilostazol</keyword>
  <keyword>progenitor cells</keyword>
  <keyword>cardiovascular disease</keyword>
  <keyword>risk factors</keyword>
  <keyword>angiogenesis</keyword>
  <keyword>biological markers</keyword>
  <keyword>endothelial function</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Cardiovascular Diseases</mesh_term>
  </condition_browse>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Cilostazol</mesh_term>
  </intervention_browse>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT02194686
org study id: B-BR-101-121
secondary id: NCKUH-10203022
nct id: NCT02194686
lead sponsor: 
collaborator: 
has dmc: No
1. The number and function of circulating endothelial progenitor cells (EPCs) are inversely
           associated with coronary risk factors and atherosclerotic diseases.

        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 high risk for cardiovascular disease.
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
allocation: Randomized
intervention model: Parallel Assignment
primary purpose: Treatment
masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
measure: Circulating EPCs Number
time frame: 3 months
description: 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.
measure: Viability (Proliferation) of EPCs
time frame: 3 months
description: 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.
measure: FMD of the Brachial Artery
time frame: 3 months
description: FMD in response to reactive hyperemia is measured in the left brachial artery in a quiet, temperature-controlled room after 10 min of bed rest. A high-resolution ultrasound machine equipped with a 7.5 mega Hertz linear array probe is used for the study. Arterial diameters are measured at the baseline and during reactive hyperemia. FMD is calculated as the percentage change in diameter compared with the baseline.
arm group label: Cilostazol
arm group type: Active Comparator
description: One tablet (100 mg) twice per day for 12 weeks
arm group label: Dummy Placebo
arm group type: Placebo Comparator
description: One tablet twice per day for 12 weeks
intervention type: Drug
intervention name: Cilostazol
description: One tablet (100 mg) twice per day for 12 weeks
arm group label: Cilostazol
other name: Pletaal (brand name)
intervention type: Drug
intervention name: Dummy Placebo
description: One tablet twice per day for 12 weeks
arm group label: Dummy Placebo
other name: Placebo
other name: Control
criteria: 
gender: All
minimum age: 20 Years
maximum age: N/A
healthy volunteers: No
last name: Ting-Hsing Chao, MD
role: Principal Investigator
affiliation: National Cheng-Kung University Hospital
facility: 
country: Taiwan
citation: Chao TH, Tseng SY, Li YH, Liu PY, Cho CL, Shi GY, Wu HL, Chen JH. A novel vasculo-angiogenic effect of cilostazol mediated by cross-talk between multiple signalling pathways including the ERK/p38 MAPK signalling transduction cascade. Clin Sci (Lond). 2012 Aug 1;123(3):147-59. doi: 10.1042/CS20110432.
PMID: 22339730
citation: Biscetti F, Pecorini G, Straface G, Arena V, Stigliano E, Rutella S, Locatelli F, Angelini F, Ghirlanda G, Flex A. Cilostazol promotes angiogenesis after peripheral ischemia through a VEGF-dependent mechanism. Int J Cardiol. 2013 Aug 10;167(3):910-6. doi: 10.1016/j.ijcard.2012.03.103. Epub 2012 Apr 2.
PMID: 22473072
citation: Chao TH, Tseng SY, Chen IC, Tsai YS, Huang YY, Liu PY, Ou HY, Li YH, Wu HL, Cho CL, Tsai LM, Chen JH. Cilostazol enhances mobilization and proliferation of endothelial progenitor cells and collateral formation by modifying vasculo-angiogenic biomarkers in peripheral arterial disease. Int J Cardiol. 2014 Mar 15;172(2):e371-4. doi: 10.1016/j.ijcard.2013.12.295. Epub 2014 Jan 11. No abstract available.
PMID: 24439864
responsible party type: Sponsor
mesh term: Cardiovascular Diseases
mesh term: Cilostazol