<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/NCT04450212</url>
  </required_header>
  <id_info>
    <org_study_id>STUDY00001593</org_study_id>
    <secondary_id>P01GM116691-02S1</secondary_id>
    <nct_id>NCT04450212</nct_id>
  </id_info>
  <brief_title>Vitamin K Supplementation Study in Healthy Volunteers (Aim 2B)</brief_title>
  <official_title>Vitamin K Supplementation Study in Healthy Volunteers (Aim 2B)</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>University of Washington</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>National Institute of General Medical Sciences (NIGMS)</agency>
      <agency_class>NIH</agency_class>
    </collaborator>
  </sponsors>
  <source>University of Washington</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
    <is_fda_regulated_drug>No</is_fda_regulated_drug>
    <is_fda_regulated_device>No</is_fda_regulated_device>
    <is_us_export>No</is_us_export>
  </oversight_info>
  <brief_summary>
    <textblock>
      The overall purpose of this study is to determine how variation in the CYP4F2 gene modulates&#xD;
      the synthesis of vitamin K-dependent clotting factors. We propose that the CYP4F2*3 gene&#xD;
      variant increases short- and long-term vitamin K concentrations in the liver by reducing the&#xD;
      efficiency of vitamin K metabolism. The investigators will study the effect of vitamin K&#xD;
      supplementation on two biomarkers of hepatic vitamin K concentration in groups with defined&#xD;
      CYP4F2*3 genotype. Specifically, the investigators will test for an association between our&#xD;
      novel biomarkers of long-term (plasma Factor II proteoforms) and short-term (urinary K-Acid&#xD;
      catabolites) hepatic vitamin K concentration and CYP4F2*3 following a 10-day period of&#xD;
      vitamin K supplementation in healthy volunteers.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      The investigators will study the effect of vitamin K supplementation on two biomarkers of&#xD;
      hepatic vitamin K concentration in groups with defined CYP4F2*3 genotype. Specifically, the&#xD;
      investigators will test for an association between our novel biomarkers of long-term (plasma&#xD;
      Factor II proteoforms) and short-term (urinary K-Acid catabolites) hepatic vitamin K&#xD;
      concentration and CYP4F2*3 following a 10-day period of vitamin K supplementation in healthy&#xD;
      volunteers.&#xD;
&#xD;
      The investigators will recruit, by posted advertisements at UWMC, male and female healthy&#xD;
      volunteers. A two-step selection process will be employed. Subjects will self-select by&#xD;
      responding to flyers, and contact the research coordinator. The research coordinator will&#xD;
      screen them for eligibility over the phone and if eligible, make an appointment for the first&#xD;
      study visit. For the first phase, the Research Coordinator will collect a buccal swab of DNA&#xD;
      from ~ 200 eligible candidates; the DNA will be tested for absence or presence of the&#xD;
      CYP4F2*3 variant allele. The PI and Research Coordinator will review the genotyping results&#xD;
      and then select 14 individuals with either a homozygous CYP4F2*3 genotype or a heterozygous&#xD;
      CYP4F2*1/*3 genotype, and a demographically matched group of 14 with a homozygous CYP4F2*1&#xD;
      genotype at the diagnostic locus for the supplementation phase of the study. The study&#xD;
      coordinator will then contact these subjects by phone for Phase II participation.&#xD;
&#xD;
      Procedures - Phase I, Buccal Swab Collection for DNA Isolation.&#xD;
&#xD;
      Demographic Questionnaire: Self-reported heritage, age, and sex will be collected through a&#xD;
      brief demographic questionnaire.&#xD;
&#xD;
      Buccal Swab: We will collect cheek cells with a cotton swab and isolate DNA and test for the&#xD;
      CYP4F2*1 and CYP4F2*3 alleles.&#xD;
&#xD;
      Genotyping: CYP4F2 genotype will be determined by a validated TaqMan assay (ABI/ThermoFisher&#xD;
      Scientific), using commercially available DNA hybridization probes to test for the absence or&#xD;
      presence of the CYP4F2*3 allele. Subjects who are either a homozygous CYP4F2*3 genotype or a&#xD;
      heterozygous CYP4F2*1/*3 genotype, and CYP4F2*3 alleles will be eligible for Phase II.&#xD;
&#xD;
      Procedures - Phase II, Vitamin K Supplementation.&#xD;
&#xD;
      Vitamin K Supplementation: Research participants selected for the supplementation study based&#xD;
      on CYP4F2 genotype will be given 1-mg/day phylloquinone (Vitacost; Natures Life K-1&#xD;
      Phylloquinone) for 10 consecutive days. Each dose will be taken in the morning (~ 8 am), with&#xD;
      one half pint of 2% milk to facilitate absorption.&#xD;
&#xD;
      Sample Collection: For the supplementation study, a venous blood sample (10 mL EDTA tube) and&#xD;
      a spot urine sample will be collected after an overnight fast from d1 through d5 and on d8&#xD;
      and d10 of Vitamin K supplementation for 10 days. Plasma will be isolated and both samples&#xD;
      will be stored at -70°C until analysis. Women of child-bearing potential will have a urine&#xD;
      pregnancy test of d1 and if positive, will be withdrawn from the study, and all samples and&#xD;
      data will be destroyed.&#xD;
&#xD;
      Measurement of Plasma Factor II Proteoforms: Plasma concentration of the 11 individual&#xD;
      proteoforms of Factor II will be measured byLC-MS/MS. This data will be used to calculate&#xD;
      three metrics of long-term vitamin K status, unFII, ucFII:iFII and inFII:aFII. These&#xD;
      represent different mathematical measures of the degree of Factor II undercarboxylation.&#xD;
      Prospective analysis of plasma vitamin K1, PIVKA II and ucOC is not planned, but will be held&#xD;
      in reserve should the need for that data arise.&#xD;
&#xD;
      Measurement of Urinary K-Acid I and II: Quantitation of K-Acid I and II, and creatinine (Cr)&#xD;
      in spot urine samples will be performed, using a validated LC-MS/MS assay that the&#xD;
      investigators have recently developed. The total and individual urinary K-Acid/Cr ratio will&#xD;
      be computed.&#xD;
&#xD;
      Statistical Analysis Plan: The investigators will conduct an unpaired t-test to compare the&#xD;
      mean response to vitamin K supplementation in the two CYP4F2 genotype groups. Response is&#xD;
      defined as the area under the vitamin K status value x time curve, corrected for baseline,&#xD;
      for the 10-day intervention period. A secondary metric will be the absolute peak change in&#xD;
      the vitamin K status ratio. Both short (urinary K-Acid/Cr ratio) and long-term (ucFII:iFII&#xD;
      and inFII:aFII) biomarkers will tested separately. Based on pilot data, a sample size of 14&#xD;
      in each genotype group will provide a power of at least 80% to detect a significant effect at&#xD;
      the significance 0.05 level, when there is a two-fold difference in the response to&#xD;
      supplementation.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Recruiting</overall_status>
  <start_date type="Actual">August 25, 2017</start_date>
  <completion_date type="Anticipated">July 31, 2023</completion_date>
  <primary_completion_date type="Anticipated">July 31, 2023</primary_completion_date>
  <phase>N/A</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Non-Randomized</allocation>
    <intervention_model>Parallel Assignment</intervention_model>
    <intervention_model_description>This is a two-phase study. Healthy adults are screened by genetic analysis in phase I and two sub-groups with specific CYP4F2 genotypes (*1/*1 and *1/*3 or *3/*3) are selected to receive vitamin K supplementation in phase II.</intervention_model_description>
    <primary_purpose>Basic Science</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Short-term Hepatic Vitamin K Status-1</measure>
    <time_frame>10-days</time_frame>
    <description>urinary vitamin K-acid/creatinine ratio</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Long-term Hepatic Vitamin K Status</measure>
    <time_frame>10-days</time_frame>
    <description>plasma factor II carboxylation state</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Short-term Hepatic Vitamin K Status-2</measure>
    <time_frame>10-days</time_frame>
    <description>Plasma vitamin K acid concentration</description>
  </secondary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Anticipated">200</enrollment>
  <condition>Vitamin K Status</condition>
  <arm_group>
    <arm_group_label>Phase I, Buccal Swab Collection for DNA Isolation</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>Approximately 200 healthy volunteers recruited. They complete a brief demographic survey and a undergo one-time buccal swab for collection of cheek cells for DNA analysis.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Phase II, Vitamin K (Vitacost) Supplementation</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>Subjects from Phase I with a homozygous CYP4F2*1 (n=14) or CYP4F2*3 carriers (n=14) are selected to receive daily vitamin K supplementation, for 10-days. Blood and urine samples are collected sequentially, at baseline, and during the supplementation period.</description>
  </arm_group>
  <intervention>
    <intervention_type>Genetic</intervention_type>
    <intervention_name>Phase I, Buccal Swab Collection for DNA Isolation</intervention_name>
    <description>Buccal swab will be taken from study participants for DNA isolation.</description>
    <arm_group_label>Phase I, Buccal Swab Collection for DNA Isolation</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Phase II, Vitamin K (Vitacost) Supplementation</intervention_name>
    <description>1-mg/day phylloquinone (Vitacost; Natures Life K-1 Phylloquinone) for 10 consecutive days. Each dose will be taken in the morning (~ 8 am), with one half pint of 2% milk to facilitate absorption. Blood and urine samples collection before and during the vitamin K supplementation period.</description>
    <arm_group_label>Phase II, Vitamin K (Vitacost) Supplementation</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          1. Participants will be males or females 18 years and older.&#xD;
&#xD;
          2. Participants must read and understand English.&#xD;
&#xD;
          3. Participants must be able to provide informed consent.&#xD;
&#xD;
          4. Women not currently pregnant or lactating.&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          1. Participants less than 18 years of age.&#xD;
&#xD;
          2. Participants unable to read and understand English.&#xD;
&#xD;
          3. Participants unable to provide informed consent.&#xD;
&#xD;
          4. Women who are pregnant or lactating.&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Kenneth E Thummel, PhD</last_name>
    <role>Principal Investigator</role>
    <affiliation>University of Washington</affiliation>
  </overall_official>
  <overall_contact>
    <last_name>Kenneth E Thummel, PhD</last_name>
    <phone>206-543-0819</phone>
    <email>thummel@uw.edu</email>
  </overall_contact>
  <location>
    <facility>
      <name>University of Washington</name>
      <address>
        <city>Seattle</city>
        <state>Washington</state>
        <zip>98195</zip>
        <country>United States</country>
      </address>
    </facility>
    <status>Recruiting</status>
    <contact>
      <last_name>Kenneth E. Thummel, PhD</last_name>
      <phone>206-543-0819</phone>
      <email>thummel@uw.edu</email>
    </contact>
  </location>
  <location_countries>
    <country>United States</country>
  </location_countries>
  <verification_date>June 2022</verification_date>
  <study_first_submitted>June 22, 2020</study_first_submitted>
  <study_first_submitted_qc>June 25, 2020</study_first_submitted_qc>
  <study_first_posted type="Actual">June 29, 2020</study_first_posted>
  <last_update_submitted>June 9, 2022</last_update_submitted>
  <last_update_submitted_qc>June 9, 2022</last_update_submitted_qc>
  <last_update_posted type="Actual">June 13, 2022</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>University of Washington</investigator_affiliation>
    <investigator_full_name>Kenneth Thummel</investigator_full_name>
    <investigator_title>Professor: Pharmaceutics</investigator_title>
  </responsible_party>
  <keyword>vitamin K</keyword>
  <keyword>pharmacogenetics</keyword>
  <keyword>blood clotting factors</keyword>
  <keyword>CYP4F2</keyword>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Vitamin K</mesh_term>
  </intervention_browse>
  <patient_data>
    <sharing_ipd>No</sharing_ipd>
  </patient_data>
  <!-- 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/NCT04450212
org study id: STUDY00001593
secondary id: P01GM116691-02S1
nct id: NCT04450212
lead sponsor: 
collaborator: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
is us export: No
The overall purpose of this study is to determine how variation in the CYP4F2 gene modulates
      the synthesis of vitamin K-dependent clotting factors. We propose that the CYP4F2*3 gene
      variant increases short- and long-term vitamin K concentrations in the liver by reducing the
      efficiency of vitamin K metabolism. The investigators will study the effect of vitamin K
      supplementation on two biomarkers of hepatic vitamin K concentration in groups with defined
      CYP4F2*3 genotype. Specifically, the investigators will test for an association between our
      novel biomarkers of long-term (plasma Factor II proteoforms) and short-term (urinary K-Acid
      catabolites) hepatic vitamin K concentration and CYP4F2*3 following a 10-day period of
      vitamin K supplementation in healthy volunteers.
The investigators will study the effect of vitamin K supplementation on two biomarkers of
      hepatic vitamin K concentration in groups with defined CYP4F2*3 genotype. Specifically, the
      investigators will test for an association between our novel biomarkers of long-term (plasma
      Factor II proteoforms) and short-term (urinary K-Acid catabolites) hepatic vitamin K
      concentration and CYP4F2*3 following a 10-day period of vitamin K supplementation in healthy
      volunteers.

      The investigators will recruit, by posted advertisements at UWMC, male and female healthy
      volunteers. A two-step selection process will be employed. Subjects will self-select by
      responding to flyers, and contact the research coordinator. The research coordinator will
      screen them for eligibility over the phone and if eligible, make an appointment for the first
      study visit. For the first phase, the Research Coordinator will collect a buccal swab of DNA
      from ~ 200 eligible candidates; the DNA will be tested for absence or presence of the
      CYP4F2*3 variant allele. The PI and Research Coordinator will review the genotyping results
      and then select 14 individuals with either a homozygous CYP4F2*3 genotype or a heterozygous
      CYP4F2*1/*3 genotype, and a demographically matched group of 14 with a homozygous CYP4F2*1
      genotype at the diagnostic locus for the supplementation phase of the study. The study
      coordinator will then contact these subjects by phone for Phase II participation.

      Procedures - Phase I, Buccal Swab Collection for DNA Isolation.

      Demographic Questionnaire: Self-reported heritage, age, and sex will be collected through a
      brief demographic questionnaire.

      Buccal Swab: We will collect cheek cells with a cotton swab and isolate DNA and test for the
      CYP4F2*1 and CYP4F2*3 alleles.

      Genotyping: CYP4F2 genotype will be determined by a validated TaqMan assay (ABI/ThermoFisher
      Scientific), using commercially available DNA hybridization probes to test for the absence or
      presence of the CYP4F2*3 allele. Subjects who are either a homozygous CYP4F2*3 genotype or a
      heterozygous CYP4F2*1/*3 genotype, and CYP4F2*3 alleles will be eligible for Phase II.

      Procedures - Phase II, Vitamin K Supplementation.

      Vitamin K Supplementation: Research participants selected for the supplementation study based
      on CYP4F2 genotype will be given 1-mg/day phylloquinone (Vitacost; Natures Life K-1
      Phylloquinone) for 10 consecutive days. Each dose will be taken in the morning (~ 8 am), with
      one half pint of 2% milk to facilitate absorption.

      Sample Collection: For the supplementation study, a venous blood sample (10 mL EDTA tube) and
      a spot urine sample will be collected after an overnight fast from d1 through d5 and on d8
      and d10 of Vitamin K supplementation for 10 days. Plasma will be isolated and both samples
      will be stored at -70°C until analysis. Women of child-bearing potential will have a urine
      pregnancy test of d1 and if positive, will be withdrawn from the study, and all samples and
      data will be destroyed.

      Measurement of Plasma Factor II Proteoforms: Plasma concentration of the 11 individual
      proteoforms of Factor II will be measured byLC-MS/MS. This data will be used to calculate
      three metrics of long-term vitamin K status, unFII, ucFII:iFII and inFII:aFII. These
      represent different mathematical measures of the degree of Factor II undercarboxylation.
      Prospective analysis of plasma vitamin K1, PIVKA II and ucOC is not planned, but will be held
      in reserve should the need for that data arise.

      Measurement of Urinary K-Acid I and II: Quantitation of K-Acid I and II, and creatinine (Cr)
      in spot urine samples will be performed, using a validated LC-MS/MS assay that the
      investigators have recently developed. The total and individual urinary K-Acid/Cr ratio will
      be computed.

      Statistical Analysis Plan: The investigators will conduct an unpaired t-test to compare the
      mean response to vitamin K supplementation in the two CYP4F2 genotype groups. Response is
      defined as the area under the vitamin K status value x time curve, corrected for baseline,
      for the 10-day intervention period. A secondary metric will be the absolute peak change in
      the vitamin K status ratio. Both short (urinary K-Acid/Cr ratio) and long-term (ucFII:iFII
      and inFII:aFII) biomarkers will tested separately. Based on pilot data, a sample size of 14
      in each genotype group will provide a power of at least 80% to detect a significant effect at
      the significance 0.05 level, when there is a two-fold difference in the response to
      supplementation.
allocation: Non-Randomized
intervention model: Parallel Assignment
intervention model description: This is a two-phase study. Healthy adults are screened by genetic analysis in phase I and two sub-groups with specific CYP4F2 genotypes (*1/*1 and *1/*3 or *3/*3) are selected to receive vitamin K supplementation in phase II.
primary purpose: Basic Science
masking: None (Open Label)
measure: Short-term Hepatic Vitamin K Status-1
time frame: 10-days
description: urinary vitamin K-acid/creatinine ratio
measure: Long-term Hepatic Vitamin K Status
time frame: 10-days
description: plasma factor II carboxylation state
measure: Short-term Hepatic Vitamin K Status-2
time frame: 10-days
description: Plasma vitamin K acid concentration
arm group label: Phase I, Buccal Swab Collection for DNA Isolation
arm group type: Experimental
description: Approximately 200 healthy volunteers recruited. They complete a brief demographic survey and a undergo one-time buccal swab for collection of cheek cells for DNA analysis.
arm group label: Phase II, Vitamin K (Vitacost) Supplementation
arm group type: Experimental
description: Subjects from Phase I with a homozygous CYP4F2*1 (n=14) or CYP4F2*3 carriers (n=14) are selected to receive daily vitamin K supplementation, for 10-days. Blood and urine samples are collected sequentially, at baseline, and during the supplementation period.
intervention type: Genetic
intervention name: Phase I, Buccal Swab Collection for DNA Isolation
description: Buccal swab will be taken from study participants for DNA isolation.
arm group label: Phase I, Buccal Swab Collection for DNA Isolation
intervention type: Dietary Supplement
intervention name: Phase II, Vitamin K (Vitacost) Supplementation
description: 1-mg/day phylloquinone (Vitacost; Natures Life K-1 Phylloquinone) for 10 consecutive days. Each dose will be taken in the morning (~ 8 am), with one half pint of 2% milk to facilitate absorption. Blood and urine samples collection before and during the vitamin K supplementation period.
arm group label: Phase II, Vitamin K (Vitacost) Supplementation
criteria: 
gender: All
minimum age: 18 Years
maximum age: N/A
healthy volunteers: Accepts Healthy Volunteers
last name: Kenneth E Thummel, PhD
role: Principal Investigator
affiliation: University of Washington
last name: Kenneth E Thummel, PhD
phone: 206-543-0819
email: thummel@uw.edu
facility: 
status: Recruiting
contact: 
country: United States
responsible party type: Principal Investigator
investigator affiliation: University of Washington
investigator full name: Kenneth Thummel
investigator title: Professor: Pharmaceutics
mesh term: Vitamin K
sharing ipd: No