Methods
Study design and dosing
The study protocol was approved by the relevant health authority in each country and by an institutional review board or an independent ethics committee at each site. Signed informed consent was obtained from each patient. The study was carried out in accordance with the principles of the International Council for Harmonisation Guidelines for Good Clinical Practice, the Declaration of Helsinki and all applicable national and local laws. This study is registered on ClinicalTrials.gov (NCT02472795).
This multicentre, double-blind, randomised, placebo-controlled 12-week study was conducted at 18 centres across Belarus, Bulgaria, Georgia, Russia, Ukraine and the USA. The study had two parts, part A and part B, which had the same study design: a 30-day screening period followed by a 12-week treatment period, a 6-week follow-up visit, and two telephone calls at 11 and 16 weeks after treatment discontinuation.
In part A, eligible patients were randomly assigned (1:1:1:1) to once daily oral administration of cenerimod 0.5, 1, 2 mg or placebo. After all patients had completed 4 weeks of treatment during part A, an Independent Data Monitoring Committee reviewed non-blinded data in an interim analysis to evaluate the safety profile of cenerimod and recommend whether the study could proceed to part B as planned (study design: online supplementary file 1). In part B, additional patients were randomised (3:1) to once daily oral administration of cenerimod 4 mg or placebo.
Randomisation was done using an interactive response technology system. The investigator, study site personnel, patients and sponsor personnel involved in the conduct of the study remained blinded to both the treatment allocation and to the interim analysis results until study closure.
Patients
Patients were eligible for inclusion in the study if they met the following criteria: were aged 18–65 years; fulfilled at least four of the American College of Rheumatology revised diagnostic criteria for SLE;21 were diagnosed at least 6 months before screening; had an SLE Disease Activity Index-2000 (SLEDAI-2K) score of at least two points for musculoskeletal or mucocutaneous manifestations; a history of, or positive serum test at screening for, antinuclear antibodies or anti-double-stranded DNA (dsDNA) antibodies; and were receiving background SLE medication (non-steroidal anti-inflammatory drugs, corticosteroids, antimalarials, mycophenolate mofetil, azathioprine or methotrexate) at stable doses for at least 30 days before randomisation. Patients were ineligible if they had severe lupus disease activity (SLEDAI-2K score >12 points), active lupus nephritis, central nervous system lupus or lupus vasculitis within 90 days prior to randomisation. Additional inclusion and exclusion criteria are described in online supplementary file 2.
Procedures
During the study, assessments were conducted at seven scheduled visits (baseline; day 1; weeks 2, 4 and 8 of treatment; at end of treatment (EOT; week 12) and at end of study (EOS; 6 weeks after EOT)). Additionally, patients were contacted via telephone 11 and 16 weeks after EOT to collect safety information, including serious adverse events (SAEs) and pregnancy status.
Assessments at each visit included blood sampling for haematology, clinical chemistry and biomarker analyses; safety and tolerability assessments (monitoring adverse events (AEs; coded using the Medical Dictionary for Regulatory Activities, version 19.0), vital signs, 12-lead electrocardiograms (ECGs), ophthalmic examinations (apart from day 1) and spirometry (apart from day 1) for forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC)); and disease activity assessments (apart from week 2) using the modified SLEDAI-2K (mSLEDAI-2K) to exclude leucopenia because of the mode of action of cenerimod.
During the treatment period, blood samples were collected before dosing at weeks 2, 4, 8 and 12, and at EOS. Trough plasma concentrations (Ctrough) of cenerimod were determined using a validated liquid chromatography coupled to tandem mass spectrometry assay with a lower limit of quantification of 0.1 ng/mL.
Predefined day 1 safety assessments included heart rate monitoring and changes in 12-lead ECG variables (including heart rate and PR, QRS and QT intervals). These assessments were performed prior to the first dose and then hourly for 6 hours. Day 1 heart rate discharge criteria were the following: ECG-derived resting heart rate more than 45 beats per minute (bpm), and if heart rate was less than 50 bpm it could not be the lowest value post dose; systolic blood pressure (BP) more than 90 mm Hg; QT interval corrected by Fredericia's formula <500 ms; no persistent ECG abnormality (eg, atrioventricular block second degree or higher) or ongoing AE requiring continued hospitalisation. In addition, 24-hour Holter ECG values were assessed. Safety areas of interest, known through clinical experience to be a class effect of S1P receptor modulators, included: cardiovascular effects including heart rate (on day 1), PR interval, systolic and diastolic BP, pulmonary function, immunomodulation including malignancies and infections, macular oedema, liver function test elevation and teratogenicity. The predefined stopping criteria for safety areas of interest are summarised in online supplementary file 3.
Outcome measures
The primary PD endpoint was change in total lymphocyte count from baseline to EOT. Other study endpoints were changes in total lymphocyte count from baseline to each on-treatment assessment and at EOS; treatment-emergent AEs (TEAEs); SAEs; AEs of special interest (AESIs; (defined as per safety area of interest) to include the anticipated risks of treatment with cenerimod, known class effects, or the events related to SLE comorbidities (eg, cardiovascular AEs)); AEs representing a clinical manifestation of an SLE flare (in the investigator’s opinion); and AEs leading to treatment discontinuation (list of AESIs and additional safety endpoints: online supplementary file 4).
Exploratory evaluations of disease activity included changes from baseline to each post-baseline asse ssment in the mSLEDAI-2K score and in the mucocutaneous and/or musculoskeletal SLEDAI-2K subscore. Exploratory biomarker endpoints included changes in anti-dsDNA and blood lymphocyte subsets from baseline to EOT and EOS. Additional exploratory endpoints are described in online supplementary file 5.
Statistical analysis
Based on assumptions from phase I study results,20 a sample size of 64 patients (12 in each cenerimod dose group and 16 in the placebo group) was deemed adequate to provide an average power of at least 90% to show a significant dose–response relationship on lymphocyte count at a one-sided alpha significance level of 5%. This assumed no lymphocyte count reduction from baseline for placebo, and a maximum 70% reduction for any cenerimod dose. For statistical analysis, all four cenerimod treatment groups were compared with combined data from the placebo groups in parts A and B.
To assess any change in total lymphocyte count, an optimised contrast test according to the Multiple Comparison Procedure and Modelling (MCP-Mod) approach22 for each considered dose–response model was used. Five models were prespecified for consideration in MCP-Mod analyses: maximum effect (Emax) curves with 50% of the effective dose (ED50) at 0.2, 0.4 and 1 mg, quadratic curve with Emax at 3 mg and sigmoid-Emax curve with ED50 at 0.4 mg and ED95 at 2 mg. Multiplicity-adjusted p-values were calculated using the Dunnett’s test. PD effects were analysed based on pairwise comparisons of reduction in total lymphocyte count from baseline for each cenerimod dose level with placebo, using an analysis of covariance (ANCOVA) model, adjusted for baseline total lymphocyte count. Statistical testing was based on a two-sided significance level of 5%.
Exploratory analyses of the change from baseline in mSLEDAI-2K score and SLEDAI-2K mucocutaneous and/or musculoskeletal score were performed using an ANCOVA, with treatment group and baseline score as factors. Mean treatment differences for each cenerimod dose compared with placebo and their corresponding two-sided 95% CIs were provided.
In general, analyses were conducted on the full analysis set (FAS) of all 67 randomised patients. Primary PD analyses were initially performed using the PD set, which included all patients who had received study treatment for at least 21 days and had valid baseline and post-baseline total lymphocyte count data. Consequently, the PD set excluded three patients from the FAS: two from the cenerimod 1 mg group and one from the placebo group. Furthermore, Ctrough levels were discovered to be low, or below the lower limit of quantification (BLQ), in four patients randomised to the cenerimod 4 mg group, a finding incompatible with compliance with study treatment. These patients were excluded from the PD set to form a post hoc modified PD (mPD) set. Exploratory analyses on disease activity and biomarkers used the mPD set in addition to the FAS. All 67 randomised patients (ie, the FAS) reported receiving at least one dose of study treatment and were included in the safety set.