Immunological Effects From Intranasal Chitin Micro-Particles
Condition: Healthy
Study type: Interventional

Summary: 
      The purpose of the study is:

        -  To investigate whether chitin can effect healthy adults, the nasal mucosa in a direction
           that will enhance the immune response to infection

        -  To investigate whether chitin influence of the nasal mucosa is well tolerated, and that
           there is no inflammation, as it seen with exposure to endotoxin.
    

Detailed description: 
      STUDY SYNOPSIS Objectives

      Primary

      •To assess the effects of intranasal dosing of chitin micro-particles on nasal mucosa
      responses with respect to the concentration of cytokines (TNF-alfa, IFN-gamma, IL-1b, IL-4
      IL5, IL, 6, IL-8, IL-10 and IL-13) in nasal lavage fluid compared to dosing with placebo.

      Secondary

        -  To assess the effects of intranasal dosing of chitin micro-particles on nasal mucosa
           responses with respect to the concentration of cytokines (TNF-alfa, IFN-gamma, IL-1b,
           IL-4 IL5, IL, 6, IL-8, IL-10 and IL-13) in nasal lavage fluid compared to dosing with
           active control.

        -  To assess the total eosinophil counts and cysteinyl leukotriene concentration in nasal
           lavage fluid at 4 and 8 hours post-dosing compared to pre-dosing.

        -  To assess the total nasal symptom scores at 4 and 8 hours post-dosing.

        -  To assess the safety and tolerability of chitin micro-particles dosed intranasally.

      Study design

      This study is a single-dose, doubled-blind placebo and single-blind active controlled
      3-period cross-over study with a 2-week washout in between each treatment. The subjects will
      be divided into 2-groups for the study and randomised to receive a single-dose of either
      intranasal chitin micro-particles or placebo for the first two periods and a single dose of
      an active control article, lipopolysaccharide (LPS), for the third period. The treatment for
      the first two periods will be double-blinded and the treatment for the third period with LPS
      will be single-blinded.

      Each single dose application for chitin micro-particles (cmp), placebo and the active control
      article, LPS, is to comprise two 0.1 ml metered sprays into each nostril. The total spray
      volume for each application is 0.4 ml equivalent to a total single-dose for chitin of 2 mg
      (0.5 mg per 0.1 ml spray), and a total single-dose for LPS, of 2 mg of endotoxin from
      Enterobacter agglomerans (0.5 mg per 0.1 ml spray).

      The climate chamber where the dosing is to be performed will run at 22°C with 3.5-4 air
      changes per hour with clean HEPA-filtered air at a relative humidity (RH) of 35-40%

      Study population

      The study will involve 12-healthy subjects conforming to the inclusion criteria randomised to
      the treatments. The duration of each subject's participation will be c.42-days with eight
      visits including screening and safety follow-up.

      Treatment plan and methods The study is to comprise three cross-over treatment periods, where
      each subject (12 in total) will receive single-doses of cmp, placebo or LPS (active control)
      in each of the three treatment periods. The study has been designed such that LPS is to be
      dosed in the last period so as to mitigate any potential issues with carry-over.

      Screening is to take place in a period of c.2-6-weeks prior to dosing. After obtaining
      informed consent, each subject will undergo a screening visit in order to review the
      inclusion/exclusion criteria and confirm eligibility. Each subject will then be assigned to
      one of two dosing groups (6 subjects per group) for each of the three study periods in order
      to co-ordinate dosing at the Clinic. The initial screening will identify 4-8 additional
      subjects (total up to 20 screened) who will be included only if one of the originally
      randomized subjects drop out of the study. Subjects that are replaced will be randomized for
      treatment and will be required to undergo all three treatment periods.

      For period 1, on day 0, the first dosing group will be randomized to receive either cmp or
      placebo. Subjects will be admitted to the Clinic at 8:30h to acclimatize, receive pre-dosing
      clinical assessment and confirm eligibility. Dosing is to start at 10:00h with assessment
      conducted in accordance with the procedures defined below. Subjects will remain in the Clinic
      until c. 20:00h. On the following day (day 1), the same subjects will receive a telephone
      interview safety follow-up. On day 2, the second dosing group will be randomized to receive
      either cmp or placebo, again receiving a telephone interview safety follow-up on following
      day (day 3). For period 2, on day 14, the first dosing group will receive the dose not
      previously assigned, either cmp or placebo and will receive a telephone interview safety
      follow-up the following day (day 15). On day 16, the second dosing group will also receive
      the dose not previously assigned, either cmp or placebo and again will receive a telephone
      interview safety follow-up the following day (day 17). For period 3, LPS will be dosed to the
      first dosing group on day 28 and to the second dosing group on day 30; with a telephone
      interview safety follow-up on days 29 and 31 respectively.

      Following the last dosing visit, subjects will receive an end-of-study safety assessment by
      telephone interview on day 42 (+/- 2 days), which will include adverse events checks.

      Each subject will be required to provide 10 ml of blood for haematological assessment at
      screening and at treatment periods 1, 2 and 3. A total of 40 ml of blood will be drawn for
      each subject for the entire duration of the study.

      Pre-dosing

        1. Safety assessment in terms of physical examination, including mouth, nose and lungs

        2. Symptom questionnaire

        3. Blood drawn for haematological tests

        4. Nasal lavage by the nasal olive test with the persons sitting with their head bent
           forward for 60 seconds performed

           Post-dosing

        5. Measurement of nasal symptom scores at 4 and 8-hours

        6. Acoustic rhinometry test performed at 4 and 8-hours

        7. Nasal lavage by the nasal olive test with the persons sitting with their head bent
           forward for 60 seconds performed at 4 and 8-hours

        8. Symptom questionnaire performed at 4 and 8-hours

        9. Safety monitoring: vital signs, heart rate, blood pressure performed at 2, 6-hours

       10. Follow-up safety interview performed 24-hours after dosing (following day) which
           includes adverse events recording and concomitant medication check.

      Primary End-point Changes in levels (maximum concentration) of TNF-alfa, IFN-gamma, IL-1b,
      IL-4, IL5, IL-6, IL-8, IL-10 and IL-13 in nasal lavage samples following intranasal dosing
      with cmp compared to placebo.

      Secondary End-point Changes in levels (maximum concentration) of TNF-alfa, IFN-gamma, IL-1b,
      IL-4, IL5, IL-6, IL-8 IL-10 and IL-13 in nasal lavage samples following intranasal dosing
      with cmp compared to lipopolysaccharide active control.

      Change from baseline:

        1. the total eosinophil counts and cysteinyl leukotriene concentration in nasal lavage
           fluid following intranasal dosing with cmp

        2. total nasal symptom scores at 4 and 8-hours following intranasal dosing with cmp

      Safety will be assessed using the adverse events, vital signs and nasal acoustic rhinometry
      following dosing with cmp versus placebo.

      Statistical Assessment For the cytokines and cell counts the concentrations are measured, and
      the log-transformed data analyzed with an ANOVA model including study day and exposure as
      fixed factor and participants as a random factor. These variables are described by the
      geometric mean, coefficient of variance (CV), relative difference between exposures including
      95% confidence interval (CI), and p-value.

      Because of ceiling and flooring problems in the data, heterogeneity in the (within
      participant) variation, or violations of the assumption of normally distributed data, it is
      not appropriate to analyze all the data of the outcomes like the symptom scores (change from
      baseline) with ANOVAs. Here non-parametric tests will be applied. A Wilcoxon Rank sum test
      will be used to compare the groups.

      Subjective symptoms at baseline are described by the median and the inter quartile range. The
      data for 'change in nasal scores' are described by box-plots. The upper adjacent value in the
      plots is the largest data-point smaller than 75th percentile plus 3/2 times the inter
      quartile range. The lower adjacent value is defined in the similar way.

      To compare placebo and the cmp exposure and taking into account a possible 'learning effect'
      the data will be analysed by a standard technique taking into account the design
      (cross-over). The participants will be divided into two groups due to the two possible
      exposure orders; one group consisting of participants having been exposed to placebo exposure
      before the cmp exposure; and another group of participants having been exposed to cmp before
      the placebo exposure. The hypothesis of no difference between cmp and placebo exposure will
      be tested by comparing the two groups with respect to the differences between the first and
      the second measurement. A Wilcoxon Rank sum test is used to compare the two groups. In a
      similar way we will test the hypothesis of 'no effect of learning'.
    

Intervention name(s): Chitin Micro Particle
Intervention type(s): Other
Intervention description(s): The total spray volume for each application is 0.4 ml equivalent to a total single-dose for chitin of 2 mg (0.5 mg per 0.1 ml spray).
A total single-dose for LPS, of 2 mg of endotoxin from Enterobacter agglomerans (0.5 mg per 0.1 ml spray)
Other intervention name(s): Chitin micro-particle
Intervention model: Single Group Assignment
Masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
Primary purpose: Prevention
Keywords: Intranasal; Chitin Micro-particles; Immunological Effects; Nasal mucosa; Cytokines; Immunological Effects from Intranasal Chitin Micro-Particles