Prefatory Study to Explore Changes in Nasal Mucociliary Clearance and to Standardize Nasal Scraping Procedure

• Cigarette (Other): After enrollment, and between Visit 2 and Visit 3, the subjects will abstain from smoking for a period of 8 hours. At Visit 3, after the 8-hour smoking abstinence period, the subjects will smoke 1 single cigarette.

Primary outcomes

• Saccharin Transit Time at t0, Start of Product Use [Baseline]: Before performing the test, each subject will spend approximately 15 minutes acclimatizing to the environment of the clinic. A 1-2 mm particle of saccharin will be placed under direct vision, approximately 1 cm behind the anterior end of the inferior turbinate. The position of the subject's head should be flexed approximately 10° by visual assessment. The saccharin transit time will be determined as the elapsed time in minutes and seconds from placement of the saccharin until the time the subject perceives a sweet taste.
• Saccharin Transit Time 4 Hours After Product Use [Measured at 4 hours after product use.]: Before performing the test, each subject will spend approximately 15 minutes acclimatizing to the environment of the clinic. A 1-2 mm particle of saccharin will be placed under direct vision, approximately 1 cm behind the anterior end of the inferior turbinate. The position of the subject's head should be flexed approximately 10° by visual assessment. The saccharin transit time will be determined as the elapsed time in minutes and seconds from placement of the saccharin until the time the subject perceives a sweet taste.
• Saccharin Transit Time 8 Hours After Product Use [Measured at 8 hours after product use.]: Before performing the test, each subject will spend approximately 15 minutes acclimatizing to the environment of the clinic. A 1-2 mm particle of saccharin will be placed under direct vision, approximately 1 cm behind the anterior end of the inferior turbinate. The position of the subject's head should be flexed approximately 10° by visual assessment. The saccharin transit time will be determined as the elapsed time in minutes and seconds from placement of the saccharin until the time the subject perceives a sweet taste.
• Saccharin Transit Time 12 Hours After Product Use [Measured at 12 hours after product use.]: Before performing the test, each subject will spend approximately 15 minutes acclimatizing to the environment of the clinic. A 1-2 mm particle of saccharin will be placed under direct vision, approximately 1 cm behind the anterior end of the inferior turbinate. The position of the subject's head should be flexed approximately 10° by visual assessment. The saccharin transit time will be determined as the elapsed time in minutes and seconds from placement of the saccharin until the time the subject perceives a sweet taste.
• Concentration of Plasma Nicotine at t0, Start of Product Use [Baseline]: Nicotine plasma concentrations will be measured using validated LC/MS/MS bioanalytical method.
• Concentration of Plasma Nicotine 4 Hours After Product Use [Measured at 4 hours after product use.]: Nicotine plasma concentrations will be measured using validated LC/MS/MS bioanalytical method.
• Concentration of Plasma Nicotine 8 Hours After Product Use [Measured at 8 hours after product use.]: Nicotine plasma concentrations will be measured using validated LC/MS/MS bioanalytical method.
• Concentration of Plasma Nicotine 12 Hours After Product Use [Measured at 12 hours after product use.]: The Nicotine plasma concentrations will be measured using validated LC/MS/MS bioanalytical method.

Secondary outcomes

• Ribonucleic Acid Quantity (Right Nostril) [Nasal scraping was performed at the final clinic visit, on Day 2, before checkout from the study.]: Ribonucleic Acid quantity: concentration measured in the right nostril (RNA protect buffer) using two nasal scraping methods
• Ribonucleic Acid Quantity (Left Nostril) [Nasal scraping was performed at the final clinic visit, on Day 2, before checkout from the study.]: Ribonucleic Acid quantity: concentration measured in the left nostril (Qiazol buffer) using two nasal scraping methods
• Ribonucleic Acid Quality (Right Nostril) [Nasal scraping was performed at the final clinic visit, on Day 2, before checkout from the study.]: Ribonucleic Acid quality assessed using the RNA integrity number: measured in the right nostril (RNA protect buffer) using two nasal scraping methods. RNA integrity Number is an algorithm for assessing RNA integrity and based on a numbering system from 1 to 10, with 1 being the most degraded profile and 10 being the most intact.
• Ribonucleic Acid Quality (Left Nostril) [Nasal scraping was performed at the final clinic visit, on Day 2, before checkout from the study.]: Ribonucleic Acid quality assessed using the RNA integrity number: measured in the left nostril (Qiazol buffer) using two nasal scraping methods. RNA integrity Number is an algorithm for assessing RNA integrity and based on a numbering system from 1 to 10, with 1 being the most degraded profile and 10 being the most intact.

Sex: Male, ages: 25 Years to 45 Years, accepts healthy volunteers

Inclusion Criteria to be met at Visit 1 and Visit 2:

* Male smokers aged ≥25 to ≤45 years old.
* Male never smokers aged ≥25 to ≤45 years old.
* Subject's BMI is comprised between 18.0 kg/m2 to 32.0 kg/m2, inclusive.
* Subject is healthy, as judged by the Investigator.

Additional Inclusion Criteria to allocate subjects in one of the two groups:

* Non-menthol cigarette smokers:

  * A positive urine cotinine test (≥200 ng/mL).
  * Smoked at least 20 cigarettes per day for at least the past 5 years.
  * eCO levels >10 parts per million (ppm).
  * No plans to quit smoking in the next 3 months.
* Never smokers:

  * Subject who has smoked less than 100 cigarettes throughout their lifetime and no cigarettes in the past 3 years.
  * A negative urine cotinine test (<200 ng/mL).
  * eCO levels ≤ 5 ppm.

Exclusion Criteria:

* As per the Investigator's judgment, the subject cannot participate in the study for any reason (e.g., medical, psychiatric, and/or social reason).
* Inability to taste sweet within 60 minutes in the STT test.
* Any condition the Principal Investigator or designee has cause to believe would interfere with the procedures for upper or lower airway function. This could include, but is not limited to, nasal/septum deviations, or nasal polyps or nasal allergies.

Participant flow

Outcome results

• Saccharin Transit Time at t0, Start of Product Use (Primary), Mean, minutes: Cigarette Smokers 8.4 (4.81 to 11.89); Never Smokers 10.1 (4.05 to 16.21)
• Saccharin Transit Time 4 Hours After Product Use (Primary), Mean, minutes: Cigarette Smokers 8.0 (5.83 to 10.12); Never Smokers 7.1 (1.98 to 12.28)
• Saccharin Transit Time 8 Hours After Product Use (Primary), Mean, minutes: Cigarette Smokers 9.9 (3.25 to 16.57); Never Smokers 10.1 (3.03 to 17.11)
• Saccharin Transit Time 12 Hours After Product Use (Primary), Mean, minutes: Cigarette Smokers 8.6 (5.34 to 11.78); Never Smokers 9.0 (2.61 to 15.47)
• Concentration of Plasma Nicotine at t0, Start of Product Use (Primary), Mean, ng/mL: Cigarette Smokers 18.8 (13.83 to 23.80); Never Smokers 0.0 (0.00 to 0.00)
• Concentration of Plasma Nicotine 4 Hours After Product Use (Primary), Mean, ng/mL: Cigarette Smokers 4.6 (1.25 to 7.90); Never Smokers 0.2 (NA to NA)
• Concentration of Plasma Nicotine 8 Hours After Product Use (Primary), Mean, ng/mL: Cigarette Smokers 2.8 (0.35 to 5.26); Never Smokers 0.2 (NA to NA)
• Concentration of Plasma Nicotine 12 Hours After Product Use (Primary), Mean, ng/mL: Cigarette Smokers 3.9 (-2.24 to 9.96); Never Smokers 0.2 (NA to NA)
• Ribonucleic Acid Quantity (Right Nostril) (Secondary), Mean, ng/uL: Nasal Scraping Method 1 47.5 +/- 23.83; Nasal Scraping Method 2 44.0 +/- 43.77
• Ribonucleic Acid Quantity (Left Nostril) (Secondary), Mean, ng/uL: Nasal Scraping Method 1 46.2 +/- 36.64; Nasal Scraping Method 2 57.9 +/- 39.94
• Ribonucleic Acid Quality (Right Nostril) (Secondary), Mean, RNA Integrity Number: Nasal Scraping Method 1 8.6 +/- 1.13; Nasal Scraping Method 2 7.1 +/- 1.91
• Ribonucleic Acid Quality (Left Nostril) (Secondary), Mean, RNA Integrity Number: Nasal Scraping Method 1 2.4 +/- 0.54; Nasal Scraping Method 2 2.3 +/- 0.41

Adverse events

Time frame: Adverse events were collected over the entire study duration of up to 7 months, with individual subject participation of a maximum of 33 days from the signature of the Informed Consent Form (ICF) until the end of the safety follow-up period.

Other adverse events by term

• Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006 Jan 31;7:3. doi: 10.1186/1471-2199-7-3. PMID 16448564

• Study Protocol (PDF, 2017-05-22)
• Statistical Analysis Plan (PDF, 2017-02-24)

• Christelle Haziza, PhD, Study chair, Philip Morris Products S.A.
• Frank Lee, MD, Principal investigator, Inflamax Research - Neptune