Sputum Matrix Metalloproteinases (MMP) mRNA and Montelukast

This study has been completed.
Sponsor:
Collaborator:
Clinical Research and Trials Unit (Norfolk & Norwich University Hospital, UK)
Information provided by:
University of East Anglia
ClinicalTrials.gov Identifier:
NCT00947453
First received: July 24, 2009
Last updated: August 22, 2012
Last verified: August 2012
  Purpose

Matrix metalloproteinases (MMPs) are a group of 24 zinc containing enzymes in man. These enzymes were originally described as cleaving extracellular matrix (ECM) substrates with a predominant role in ECM homeostasis, but it is now clear that they have much wider functionality. An imbalance between MMP activity and that of their inhibitors (tissue inhibitors of metalloproteinases, TIMPs) is considered to play a critical role in the synthesis or degradation of the extracellular matrix of the airway architecture which results in fixed airflow obstruction in both asthma and chronic obstructive pulmonary disease (COPD). Using quantitative real time polymerase chain reaction (RT-PCR) the investigators have identified a difference between the level of steady state mRNA for MMP-9, MMP-14 and MMP-2 in 2 patients with asthma compared to 4 healthy controls using our method. However the investigators require further refinement of the process in order to optimise RNA quality and to evaluate the effect of montelukast across the entire family of MMPs and their inhibitors (TIMPs).


Condition Intervention Phase
Asthma
Drug: montelukast
Phase 2

Study Type: Interventional
Study Design: Endpoint Classification: Safety/Efficacy Study
Intervention Model: Single Group Assignment
Masking: Open Label
Primary Purpose: Treatment
Official Title: The Effect of Montelukast Therapy on mRNA Profile of Matrix Metalloproteinases and Their Inhibitors in the Sputum of Patients With Asthma

Resource links provided by NLM:


Further study details as provided by University of East Anglia:

Primary Outcome Measures:
  • Primary endpoint is the MMP and TIMP mRNA profile relative to a housekeeping gene [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]

Secondary Outcome Measures:
  • The difference between treatment with montelukast for 8 weeks and placebo for mRNA for MMP and TIMP [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
  • The difference between treatment with montelukast for 8 weeks and placebo for Spirometry - FEV1, FVC, FEV1/FVC ratio [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
  • The difference between treatment with montelukast for 8 weeks and placebo for Induced sputum differential cell count [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]

Enrollment: 15
Study Start Date: July 2009
Study Completion Date: December 2011
Primary Completion Date: December 2011 (Final data collection date for primary outcome measure)
Arms Assigned Interventions
Experimental: montelukast group
Identified patients with asthma to recieve Montelukast 10 mg (Merck Sharp & Dohme Ltd, Herts, UK) at 0800 am once daily for 8 weeks.
Drug: montelukast
montelukast 10 mg once daily for 8 weeks

Detailed Description:

The following measurements will be performed at screening:

  • Informed consent
  • Clinical examination
  • Spirometry
  • Induced sputum

The following will be performed after 8 weeks of study medication:

  • Clinical examination
  • Spirometry
  • Induced sputum
  • Diary Card

Spirometry:

This will be performed with a Microlab spirometer (Micro Medical Ltd, Rochester, Kent, UK). The procedure will be according to American Thoracic Society specifications(13).

Diary Card Data:

Patients will record their symptoms on a daily basis in the morning according to "cough", "breathlessness" and "wheeze" on a 4 point scale with 0=no symptoms and 3=maximal symptoms. A total symptom score will be calculated out of 12. Patients will also measure their peak expiratory flow on a daily basis in the morning and record the highest of three measurements. They will record that they have taken their study medication.

Sputum Induction & Examination:

Sputum will be obtained with hypertonic saline by the method described by Pizzichini et al(14) inhaling increasing concentrations of saline (3, 4 and 5%) each for 7 minutes, through a mouthpiece. After each period of inhalation, FEV1 will be measured for safety. Subjects will be asked to cough sputum into a sterile container. Total cell count of leukocytes will be obtained in a modified Neubauer haemocytometer. The cell viability will be determined by the trypan blue exclusion method. Four hundred non squamous cells will be counted in Wright-stained slides and the results will be expressed as a percentage and absolute number of the total non squamous count. Measurement of MMP-9, 12 TIMP-1 and TGFb will be performed in sputum supernatant.

Profile of mRNA of MMP and TIMPs:

Total RNA will be extracted from the cellular content of the induced sputum plug using a combination of Trizol extraction and Qiagen RNeasy spin columns in a similar way to previously described12. Quantitative RT-PCR, using previously developed primers and probes, will be used to determine the relative quantities of mRNA of MMPs and TIMPs as described12. We remain the only centre in the world to routinely profile the entire MMP and TIMP gene family in human samples. This gives an all encompassing view of the involvement of these enzymes and inhibitors in the disease process and also sheds light on potential new biomarkers. The possibility of expanding the gene profiling without the need for additional sputum collection also adds value to the research. This might include other proteinase families with roles in ECM breakdown or in inflammation.

  Eligibility

Ages Eligible for Study:   18 Years to 60 Years
Genders Eligible for Study:   Both
Accepts Healthy Volunteers:   No
Criteria

Inclusion Criteria:

  • Male or female, aged 18 to 60 years.
  • Diagnosed with asthma, defined as episodic chest tightness, wheezing and dyspnoea, cough.
  • Non-Smoker or Ex-Smoker for at least 10 years and a smoking history of less than 5 pack years.
  • History of asthma symptoms for more than 10years.
  • Receiving as required short acting bronchodilators.
  • Post bronchodilator FEV1 50 to 100 % predicted
  • Evidence of airway calibre reversibility within the previous 12 months: reversibility to salbutamol of 12% following 400mcg inhaled salbutamol, histamine PC20 < 8mg/ml, diurnal variation in peak expiratory flow of 20%.
  • Able to produce sputum after induction with saline.

Exclusion criteria:

  • Cardiac or pulmonary disease other than asthma.
  • Respiratory infection defined as fever, nasal/sinus congestion, fatigue, cough, antibiotic use or yellow/green sputum within 4 weeks prior to study.
  • Receiving inhaled or oral corticosteroid therapy, long acting Beta2 agonist therapy or leukotriene modifying therapy for the previous 1 month.
  • Severe or uncontrolled co-morbid disease.
  • Pregnancy or breastfeeding.
  • Unable to give written informed consent
  Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.

Please refer to this study by its ClinicalTrials.gov identifier: NCT00947453

Locations
United Kingdom
University of East Anglia
Norwich, Norfolk, United Kingdom, NR47TJ
Sponsors and Collaborators
University of East Anglia
Clinical Research and Trials Unit (Norfolk & Norwich University Hospital, UK)
Investigators
Principal Investigator: Andrew M Wilson, MRCP (UK) University of East Anglia
  More Information

Publications:

Responsible Party: Dr Andrew M Wilson, University of East Anglia
ClinicalTrials.gov Identifier: NCT00947453     History of Changes
Other Study ID Numbers: 2009RESP01, EudraCT Number: 2008-008364-27
Study First Received: July 24, 2009
Last Updated: August 22, 2012
Health Authority: United Kingdom: National Health Service

Keywords provided by University of East Anglia:
asthma
montelukast

Additional relevant MeSH terms:
Asthma
Bronchial Diseases
Respiratory Tract Diseases
Lung Diseases, Obstructive
Lung Diseases
Respiratory Hypersensitivity
Hypersensitivity, Immediate
Hypersensitivity
Immune System Diseases
Montelukast
Anti-Asthmatic Agents
Respiratory System Agents
Therapeutic Uses
Pharmacologic Actions
Leukotriene Antagonists
Hormone Antagonists
Hormones, Hormone Substitutes, and Hormone Antagonists
Physiological Effects of Drugs

ClinicalTrials.gov processed this record on October 19, 2014