<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/NCT01694381</url>
  </required_header>
  <id_info>
    <org_study_id>TS01-01</org_study_id>
    <secondary_id>2012-002225-30</secondary_id>
    <nct_id>NCT01694381</nct_id>
  </id_info>
  <brief_title>Research Into the Effect of a Clot-dissolving Agent and Its Inhibitor</brief_title>
  <official_title>Phase 1 Trial of Mutant proUK, M5, and Its Inhibitor, C1-inhibitor</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>TSI, LLC</agency>
      <agency_class>Industry</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>TSI, LLC</source>
  <oversight_info>
    <has_dmc>Yes</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Single-chain urokinase-type plasminogen activator (pro-urokinase) is a highly effective&#xD;
      thrombolytic drug. At pharmacologic concentrations however, pro-urokinase is converted to&#xD;
      urokinase - a non specific thrombolytic, limiting its therapeutic use. Mutant pro-urokinase&#xD;
      (M5) is more stable and its conversion to urokinase is inhibited by C1-inhibitor.&#xD;
&#xD;
      The primary objectives of the study are:&#xD;
&#xD;
        -  To assess the overall safety and tolerability related to systemic plasminogen activation&#xD;
           of single doses of M5 over a wide dose range (study part I).&#xD;
&#xD;
        -  To assess the effect of single doses of C1-inhibitor on the overall safety and&#xD;
           tolerability of single doses of M5 and its effect on M5-induced coagulation changes&#xD;
           (study part II).&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Mutant proUK, M5, was specifically designed to improve the plasma stability of single-chain&#xD;
      proUK by reducing its intrinsic catalytic activity and allowing it to retain its proenzyme&#xD;
      form until it encounters a thrombus. As an additional consequence of the mutation, its&#xD;
      two-chain enzymatic form (tcM5) is sensitive to inhibition by C1-inhibitor (C1INH) a&#xD;
      relatively abundant plasma inhibitor.&#xD;
&#xD;
      While it has a negligible effect on urokinase (UK), C1INH inhibits tcM5 irreversibly,&#xD;
      preventing non-specific plasminogen activation, responsible for bleeding complications. The&#xD;
      effect of endogenous C1INH can be augmented by the addition of exogenous C1INH. In vitro&#xD;
      studies in rats and dogs indicated that adding C1INH to plasma prior to clot lysis by M5&#xD;
      prevented bleeding, fibrinogenolysis and plasminogen depletion but did not affect the rate of&#xD;
      fibrinolysis. In a recent pilot rat stroke study, at similarly effective doses, M5, preceded&#xD;
      by C1INH adjunctive therapy, was equivalent to tPA alone but caused significantly less ICH,&#xD;
      was much more effective than tPA preceded by adjunctive C1INH, and was the only group with a&#xD;
      significant functional improvement at 24h.&#xD;
&#xD;
      Dose restrictions which limit efficacy of tPA-based thrombolysis are expected to be&#xD;
      circumvented by M5 preceded by adjunctive C1INH. C1INH is a commercially available plasma&#xD;
      derived product with a well-established safety and efficacy profile and is currently&#xD;
      indicated and available for routine prophylaxis of hereditary angioedema (HAE).&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Terminated</overall_status>
  <start_date type="Actual">September 2012</start_date>
  <completion_date type="Actual">October 2012</completion_date>
  <primary_completion_date type="Actual">October 2012</primary_completion_date>
  <phase>Early Phase 1</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>
    <intervention_model_description>The trial consists of two study parts in which healthy male volunteers will randomly and in a double-blinded manner receive a single dose of M5 or M5-placebo intravenously, with (part II) or without (part I) a preceding single dose of C1-inhibitor.</intervention_model_description>
    <primary_purpose>Treatment</primary_purpose>
    <masking>Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)</masking>
    <masking_description>Double-blinded study.</masking_description>
  </study_design_info>
  <primary_outcome>
    <measure>Changes to vital signs, routine safety laboratory results, or ECG-findings</measure>
    <time_frame>-42d, -14h, -15', 15', 30', 45',60', 90', 10h, 24h, 48h, 7d</time_frame>
  </primary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">8</enrollment>
  <condition>Acute Ischemic Stroke</condition>
  <arm_group>
    <arm_group_label>mutant pro-urokinase (M5) alone</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>In the first study part subjects in cohorts of 4 will receive ascending doses of either M5 (3 subjects) or M5-placebo (1 subject) without C1-inhibitor.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>In the second study part subjects in cohorts of 5 will receive ascending doses of either M5 or M5-placebo, preceded by a single intravenous dose of C1-inhibitor or C1-inhibitor-placebo. Each subject will randomly be allocated to one of the following treatment arms within one cohort:&#xD;
C1-inhibitor followed by M5 (3 subjects);&#xD;
C1-inhibitor followed by M5-placebo (1 subject);&#xD;
C1-inhibitor-placebo followed by M5-placebo (1 subject).&#xD;
Dose levels of both M5 and C1-inhibitor within each cohort will be chosen based on the available safety, pharmacokinetic and pharmacodynamic data of the preceding cohorts.</description>
  </arm_group>
  <intervention>
    <intervention_type>Drug</intervention_type>
    <intervention_name>M5</intervention_name>
    <description>single point mutant of serine protease prourokinase</description>
    <arm_group_label>Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor</arm_group_label>
    <arm_group_label>mutant pro-urokinase (M5) alone</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Drug</intervention_type>
    <intervention_name>C1-inhibitor</intervention_name>
    <description>a protease inhibitor belonging to the serpin superfamily.</description>
    <arm_group_label>Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Be male, aged between 18 and 35 years inclusive, and with a body weight of at least 60&#xD;
             kg and a body mass index (BMI) between 18.5 and 25 kg/m2 inclusive.&#xD;
&#xD;
          -  Be without clinical significant abnormalities according to the investigator's&#xD;
             judgment, based on a detailed medical history, a complete physical examination&#xD;
             (including vital signs), a standard 12-lead electrocardiogram, urinalysis, and routine&#xD;
             clinical laboratory tests.&#xD;
&#xD;
          -  Have endogenous C1-inhibitor, α2-antiplasmin, fibrinogen, and plasminogen levels&#xD;
             within the laboratory's reference range.&#xD;
&#xD;
          -  Have a negative serology for HIV, HBsAg, and HCV.&#xD;
&#xD;
          -  Have a negative test for alcohol and drugs of abuse at screening and on study day -1.&#xD;
&#xD;
          -  Be capable of understanding and willing to comply with the conditions and restrictions&#xD;
             of the protocol.&#xD;
&#xD;
          -  Have read, understood and provided written informed consent.&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Has a known or suspected inherited, congenital, or acquired disease or condition that&#xD;
             affects the haemostatic or coagulation pathways or that is associated with an&#xD;
             increased bleeding tendency.&#xD;
&#xD;
          -  Has a reasonable chance of developing a clinically significant bleeding event or a&#xD;
             bleeding event that may go undetected for a considerable amount of time during the&#xD;
             study, for example:&#xD;
&#xD;
          -  Has undergone major (internal) surgery or trauma within the last three months of the&#xD;
             anticipated dosing day;&#xD;
&#xD;
          -  Has an intestinal or cerebral vascular malformation;&#xD;
&#xD;
          -  Has participated in high impact contact sports, such as kick-boxing, within two weeks&#xD;
             of the anticipated dosing day.&#xD;
&#xD;
          -  Has received any systemically absorbed drug or substance (including prescription,&#xD;
             over-the-counter, or alternative remedies) that is not permitted by this protocol&#xD;
             prior to dosing without undergoing a wash-out period of at least seven times the&#xD;
             elimination half-life of the product. For aspirin or other products inhibiting&#xD;
             thrombocyte-aggregation the wash-out period must not be less than 28 days.&#xD;
&#xD;
          -  Has smoked tobacco in any form within three months of dosing, or has ever smoked more&#xD;
             than five cigarettes per day (or equivalent) on average.&#xD;
&#xD;
          -  Has received blood or plasma derivatives in the year preceding the administration day.&#xD;
&#xD;
          -  Has lost blood or plasma outside the limits of the local blood donation service (i.c.&#xD;
             Sanquin) three months prior to dosing.&#xD;
&#xD;
          -  Has a known hypersensitivity to any of the investigational material or related&#xD;
             compounds.&#xD;
&#xD;
          -  Has a history of severe hypersensitivity or of an allergy with severe reactions.&#xD;
&#xD;
          -  Has a history of substance abuse, including caffeine, tobacco, and alcohol.&#xD;
&#xD;
          -  Has a condition or demonstrates an attitude that in the opinion of the investigator&#xD;
             might jeopardise the subject's health or well-being, or the scientific integrity of&#xD;
             the study results.&#xD;
&#xD;
          -  Is mentally or legally incapacitated to provide informed consent.&#xD;
      </textblock>
    </criteria>
    <gender>Male</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>35 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Koos Burggraaf, MD, PhD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Center for Human Drug Research</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Center for Human Drug Research</name>
      <address>
        <city>Leiden</city>
        <zip>2333</zip>
        <country>Netherlands</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Netherlands</country>
  </location_countries>
  <reference>
    <citation>Tomasi S, Sarmientos P, Giorda G, Gurewich V, Vercelli A. Mutant prourokinase with adjunctive C1-inhibitor is an effective and safer alternative to tPA in rat stroke. PLoS One. 2011;6(7):e21999. doi: 10.1371/journal.pone.0021999. Epub 2011 Jul 14.</citation>
    <PMID>21779364</PMID>
  </reference>
  <reference>
    <citation>Pannell R, Kung W, Gurewich V. C1-inhibitor prevents non-specific plasminogen activation by a prourokinase mutant without impeding fibrin-specific fibrinolysis. J Thromb Haemost. 2007 May;5(5):1047-54. doi: 10.1111/j.1538-7836.2007.02453.x.</citation>
    <PMID>17459007</PMID>
  </reference>
  <reference>
    <citation>Gurewich V, Pannell R, Simmons-Byrd A, Sarmientos P, Liu JN, Badylak SF. Thrombolysis vs. bleeding from hemostatic sites by a prourokinase mutant compared with tissue plasminogen activator. J Thromb Haemost. 2006 Jul;4(7):1559-65. doi: 10.1111/j.1538-7836.2006.01993.x.</citation>
    <PMID>16839354</PMID>
  </reference>
  <reference>
    <citation>Liu JN, Liu JX, Liu Bf BF, Sun Z, Zuo JL, Zhang Px PX, Zhang J, Chen Yh YH, Gurewich V. Prourokinase mutant that induces highly effective clot lysis without interfering with hemostasis. Circ Res. 2002 Apr 19;90(7):757-63. doi: 10.1161/01.res.0000014825.71092.bd.</citation>
    <PMID>11964367</PMID>
  </reference>
  <verification_date>January 2021</verification_date>
  <study_first_submitted>September 24, 2012</study_first_submitted>
  <study_first_submitted_qc>September 24, 2012</study_first_submitted_qc>
  <study_first_posted type="Estimate">September 27, 2012</study_first_posted>
  <last_update_submitted>January 14, 2021</last_update_submitted>
  <last_update_submitted_qc>January 14, 2021</last_update_submitted_qc>
  <last_update_posted type="Actual">January 19, 2021</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>Acute Ischemic Stroke</keyword>
  <keyword>TS01</keyword>
  <keyword>Mutant pro-urokinase</keyword>
  <keyword>M5</keyword>
  <keyword>C1 inhibitor</keyword>
  <keyword>Safety</keyword>
  <keyword>Stroke</keyword>
  <keyword>Tolerability</keyword>
  <keyword>Ischemia</keyword>
  <keyword>Cerebral Infarction</keyword>
  <keyword>Cerebrovascular Disorders</keyword>
  <keyword>Vascular Diseases</keyword>
  <keyword>Cardiovascular Diseases</keyword>
  <keyword>Brain Infarction</keyword>
  <keyword>Brain Ischemia</keyword>
  <keyword>Fibrinolytic Agents</keyword>
  <keyword>Thrombolytic Agents</keyword>
  <keyword>Molecular Mechanisms of Pharmacological Action</keyword>
  <keyword>Pharmacologic Actions</keyword>
  <keyword>Cardiovascular Agents</keyword>
  <keyword>Therapeutic Uses</keyword>
  <keyword>Hematologic Agents</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Ischemic Stroke</mesh_term>
  </condition_browse>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Complement C1 Inhibitor Protein</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/NCT01694381
org study id: TS01-01
secondary id: 2012-002225-30
nct id: NCT01694381
lead sponsor: 
has dmc: Yes
Single-chain urokinase-type plasminogen activator (pro-urokinase) is a highly effective
      thrombolytic drug. At pharmacologic concentrations however, pro-urokinase is converted to
      urokinase - a non specific thrombolytic, limiting its therapeutic use. Mutant pro-urokinase
      (M5) is more stable and its conversion to urokinase is inhibited by C1-inhibitor.

      The primary objectives of the study are:

        -  To assess the overall safety and tolerability related to systemic plasminogen activation
           of single doses of M5 over a wide dose range (study part I).

        -  To assess the effect of single doses of C1-inhibitor on the overall safety and
           tolerability of single doses of M5 and its effect on M5-induced coagulation changes
           (study part II).
Mutant proUK, M5, was specifically designed to improve the plasma stability of single-chain
      proUK by reducing its intrinsic catalytic activity and allowing it to retain its proenzyme
      form until it encounters a thrombus. As an additional consequence of the mutation, its
      two-chain enzymatic form (tcM5) is sensitive to inhibition by C1-inhibitor (C1INH) a
      relatively abundant plasma inhibitor.

      While it has a negligible effect on urokinase (UK), C1INH inhibits tcM5 irreversibly,
      preventing non-specific plasminogen activation, responsible for bleeding complications. The
      effect of endogenous C1INH can be augmented by the addition of exogenous C1INH. In vitro
      studies in rats and dogs indicated that adding C1INH to plasma prior to clot lysis by M5
      prevented bleeding, fibrinogenolysis and plasminogen depletion but did not affect the rate of
      fibrinolysis. In a recent pilot rat stroke study, at similarly effective doses, M5, preceded
      by C1INH adjunctive therapy, was equivalent to tPA alone but caused significantly less ICH,
      was much more effective than tPA preceded by adjunctive C1INH, and was the only group with a
      significant functional improvement at 24h.

      Dose restrictions which limit efficacy of tPA-based thrombolysis are expected to be
      circumvented by M5 preceded by adjunctive C1INH. C1INH is a commercially available plasma
      derived product with a well-established safety and efficacy profile and is currently
      indicated and available for routine prophylaxis of hereditary angioedema (HAE).
allocation: Randomized
intervention model: Parallel Assignment
intervention model description: The trial consists of two study parts in which healthy male volunteers will randomly and in a double-blinded manner receive a single dose of M5 or M5-placebo intravenously, with (part II) or without (part I) a preceding single dose of C1-inhibitor.
primary purpose: Treatment
masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
masking description: Double-blinded study.
measure: Changes to vital signs, routine safety laboratory results, or ECG-findings
time frame: -42d, -14h, -15', 15', 30', 45',60', 90', 10h, 24h, 48h, 7d
arm group label: mutant pro-urokinase (M5) alone
arm group type: Experimental
description: In the first study part subjects in cohorts of 4 will receive ascending doses of either M5 (3 subjects) or M5-placebo (1 subject) without C1-inhibitor.
arm group label: Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor
arm group type: Experimental
description: In the second study part subjects in cohorts of 5 will receive ascending doses of either M5 or M5-placebo, preceded by a single intravenous dose of C1-inhibitor or C1-inhibitor-placebo. Each subject will randomly be allocated to one of the following treatment arms within one cohort:
C1-inhibitor followed by M5 (3 subjects);
C1-inhibitor followed by M5-placebo (1 subject);
C1-inhibitor-placebo followed by M5-placebo (1 subject).
Dose levels of both M5 and C1-inhibitor within each cohort will be chosen based on the available safety, pharmacokinetic and pharmacodynamic data of the preceding cohorts.
intervention type: Drug
intervention name: M5
description: single point mutant of serine protease prourokinase
arm group label: Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor
arm group label: mutant pro-urokinase (M5) alone
intervention type: Drug
intervention name: C1-inhibitor
description: a protease inhibitor belonging to the serpin superfamily.
arm group label: Mutant pro-urokinase (M5) and its inhibitor, C1 inhibitor
criteria: 
gender: Male
minimum age: 18 Years
maximum age: 35 Years
healthy volunteers: Accepts Healthy Volunteers
last name: Koos Burggraaf, MD, PhD
role: Principal Investigator
affiliation: Center for Human Drug Research
facility: 
country: Netherlands
citation: Tomasi S, Sarmientos P, Giorda G, Gurewich V, Vercelli A. Mutant prourokinase with adjunctive C1-inhibitor is an effective and safer alternative to tPA in rat stroke. PLoS One. 2011;6(7):e21999. doi: 10.1371/journal.pone.0021999. Epub 2011 Jul 14.
PMID: 21779364
citation: Pannell R, Kung W, Gurewich V. C1-inhibitor prevents non-specific plasminogen activation by a prourokinase mutant without impeding fibrin-specific fibrinolysis. J Thromb Haemost. 2007 May;5(5):1047-54. doi: 10.1111/j.1538-7836.2007.02453.x.
PMID: 17459007
citation: Gurewich V, Pannell R, Simmons-Byrd A, Sarmientos P, Liu JN, Badylak SF. Thrombolysis vs. bleeding from hemostatic sites by a prourokinase mutant compared with tissue plasminogen activator. J Thromb Haemost. 2006 Jul;4(7):1559-65. doi: 10.1111/j.1538-7836.2006.01993.x.
PMID: 16839354
citation: Liu JN, Liu JX, Liu Bf BF, Sun Z, Zuo JL, Zhang Px PX, Zhang J, Chen Yh YH, Gurewich V. Prourokinase mutant that induces highly effective clot lysis without interfering with hemostasis. Circ Res. 2002 Apr 19;90(7):757-63. doi: 10.1161/01.res.0000014825.71092.bd.
PMID: 11964367
responsible party type: Sponsor
mesh term: Ischemic Stroke
mesh term: Complement C1 Inhibitor Protein