Study Evaluating ECG Effects, Safety, Tolerability and Pharmacokinetics of Single Doses of Modufolin (Arfolitixorin) in Healthy Volunteers Tetrahydrofolate in Healthy Male Volunteers: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for Study Evaluating ECG Effects, Safety, Tolerability and Pharmacokinetics of Single Doses of Modufolin (Arfolitixorin) in Healthy Volunteers Tetrahydrofolate in Healthy Male Volunteers (NCT03203564), a randomized phase 1 study. The primary outcome is Change-from-baseline QTcF (ΔQTcF).

2. Clinical Question

This is a question about ECG measurements after dosing in healthy volunteers, rather than a question about clinical benefit in a patient population. QTcF is a heart-rate-corrected measure of the QT interval. Tracking change from baseline across scheduled post-dose assessments can characterize the timing and magnitude of ECG changes within a study. The record also lists safety, tolerability and pharmacokinetics as study aims, with secondary outcomes covering other ECG measurements and categorical ECG findings.

The registry describes randomization and triple masking, and the official title calls the study double-blind, single-center and placebo-controlled. These design descriptions are not fully consistent about masking. The registry record does not report details sufficient to resolve that discrepancy. Nor does it provide the actual QTcF estimates, their standard errors, confidence intervals or test results. Consequently, the stated question can be identified, but its answer cannot be determined from the outcome information reported in the registry record.

3. Trial Design

The official title describes an adaptive, randomized, double-blind, single-center, placebo-controlled Phase I study of single ascending doses of Modufolin in healthy male volunteers. The registry fields classify the trial as Phase I, randomized and triple-masked, while its model field says single-group. The listed arms, however, include three Modufolin dose cohorts and placebo. These differences in the record should be kept in mind rather than reconciled by assumption.

The enrollment is reported as 33. The safety figures list 8 participants at risk in each of the 200 mg/m2, 350 mg/m2 and 500 mg/m2 Modufolin groups, and 9 in the placebo group. The study dates are reported as starting on 2017-07-04, with primary completion on 2017-08-25, and the overall status is completed. The sponsor is Isofol Medical AB.

The principal assessment was change from baseline QTcF at prespecified times before and after dosing. The measurement description says that up to 10 ECG replicates were extracted at each nominal time point using TQT Plus methods from continuous Holter recordings. Participants were supinely resting for at least 10 min before ECG recording. Baseline was defined as the average of ECG intervals at the pre-dose time points 45, 30 and 15 min before dosing. The schedule also includes pre-dose assessments at -2 h and -45 min, as well as post-dose assessments from 0 through 24h.

The arm descriptions contain a dose discrepancy: the group titles identify 200 mg/m2, 350 mg/m2 and 500 mg/m2, while the description for the Modufolin arm refers to randomization to 100 mg. The record does not explain this wording. It also does not report the adaptive design rules, dose-escalation decision criteria, allocation procedure details, or the exact masking procedures. These elements therefore cannot be reconstructed from the registry record.

4. Participant Flow and Accounting

The registry reports how participants moved through each stage of the study. All counts below are exactly as posted; missing counts are not inferred from other stages.

Before assignment: Subjects were screened for eligibility according to study-specific inclusion/exclusion criteria within four weeks prior to start of study treatment (Visit 1; Screening Visit).

Overall Study

Analysis population for the primary outcome, as defined in the registry: The QT/QTc Analysis Set includes all subjects in the Safety Analysis Set with measurements at baseline as well as on-treatment with at least one post-dose time point with a valid ΔQTcF (change from baseline QTcF) value.

5. Baseline Characteristics

The table reproduces the baseline characteristics posted in the registry, which describe the participants analysed at baseline and help show whether the randomized groups were comparable.

6. Primary Endpoint

The registry lists the primary outcome as Change-from-baseline QTcF (ΔQTcF), measured over Pre-dose at following timepoints -2 h, -45 min, -30 min, -15 min. Then at following timepoints; 0, 5 min, 15 min, 30 min, 1 h, 2h, 3h, 4h, 5h, 6h, 8h, 12h and 24h post-dose..

The primary endpoint is change from baseline QTcF, denoted ΔQTcF, measured in msec. QTcF refers to a QT interval corrected for heart rate using the Fridericia correction. As a general statistical and ECG concept, correction is used because the QT interval varies with heart rate; the correction aims to make comparisons across measurements more interpretable when heart rate differs. Change from baseline expresses a post-dose measurement relative to a participant’s pre-dose reference.

The registry defines baseline as the average of ECG intervals measured 45, 30 and 15 min before dosing on Day 1. Its outcome description calls for assessments at the listed pre-dose and post-dose times, with ECG replicates extracted from continuous recordings. The primary outcome parameter is identified as a least-squares mean, with standard error as the dispersion measure. A least-squares mean is generally a model-based mean estimate; the registry record does not report the model specification or covariates used to obtain it.

Serial ECG measurements can describe how an endpoint changes over time after a dose, rather than relying on one measurement alone. QTcF is relevant because the study purpose specifically includes assessment of ECG effects. However, the registry record does not report the actual ΔQTcF estimates at any assessment time, their uncertainty intervals, or a stated comparison between groups. The measurement plan and endpoint definition are available, but the numerical findings needed to assess the observed pattern are not reported in the registry record.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

The primary measure is ΔQTcF, or change from baseline in QTcF. In general, an ECG interval is a measured duration, and heart-rate correction attempts to account for the relationship between heart rate and QT duration. QTcF names a Fridericia-corrected QT interval. The registry record gives the endpoint name and unit, msec, but does not give a detailed account of the correction algorithm beyond that label. Interpretation should therefore stay with the stated measure and should not imply that unreported processing choices are known.

What does change from baseline express?

In general, change from baseline compares a participant’s later measurement with a defined pre-treatment reference. This can help describe change over the assessment period while taking the starting measurement into account. Here, the registry defines baseline as the average of the measurements at 45, 30 and 15 min pre-dose on Day 1. Although the schedule includes an assessment at -2 h and one at -45 min, those are not included in the stated baseline definition. This distinction matters: a reader should use the registry’s explicit definition rather than assume that every pre-dose measurement contributes to baseline.

The endpoint description says that ECG measurements were collected at nominal pre-dose and post-dose times and that up to 10 ECG replicates were extracted at each time point. It describes extraction from continuous Holter recordings and a resting period before ECG recordings. Replicate ECG measurements can, as a general measurement principle, help characterize a participant’s value at a time point, but the record does not specify how replicates were combined for each endpoint estimate. It also does not give the number of valid replicates by participant or time point.

What is a least-squares mean?

As a general concept, a least-squares mean is a model-based mean, sometimes called an adjusted mean. It is calculated from a statistical model and can reflect the model’s treatment of groups and other included terms. It is not necessarily identical to a simple arithmetic average. The registry identifies the primary parameter as a least-squares mean and standard error as its dispersion, but does not report the model formula, included factors, covariance structure, or estimation method. Without these details and the estimate values, the degree of adjustment and precision cannot be assessed.

A standard error describes the estimated variability of an estimator across repeated samples under the model assumptions. It is not the spread of individual participant values. A confidence interval is a general way to communicate uncertainty around an estimate, using the estimate and its uncertainty under a stated procedure. The registry record does not provide standard-error values or confidence intervals for the primary endpoint. Neither the amount of uncertainty nor the range of plausible effects can be inferred from the study design description alone.

Why does the schedule include repeated times?

Repeated measurements can reveal whether a response is concentrated at particular times, changes over time, or differs across dose groups. This is a general advantage of serial measurement, but it also creates an analysis issue: conclusions can depend on which time points and contrasts are emphasized. The record lists multiple post-dose times through 24h and describes the primary endpoint across those times. It does not identify a single primary time point, state how comparisons across time were prioritized, or report the numerical estimates for the different assessments.

The registry also identifies a secondary outcome relating ΔΔQTc and plasma concentrations of Modufolin and metabolites. In general, a concentration-response analysis asks whether outcome changes vary with measured exposure. The record names the relationship but does not provide its estimates, model, data handling or inferential results. It therefore cannot support a conclusion about the shape, strength or precision of any concentration-response pattern.

What can be inferred from the design fields?

Randomization can, in general, reduce systematic differences between assigned groups, while masking can reduce some biases in treatment administration or outcome assessment. These are design principles, not guarantees that a study is free from bias. The record describes randomization, labels masking as triple, and calls the study double-blind in its official title. It also labels the model single-group despite listing placebo and multiple dose groups. The record does not explain these inconsistencies, the randomization sequence, allocation concealment or who was masked. Details of the adaptive design and any dose-escalation rules are also not reported in the registry record.

Finally, the adverse-event counts are descriptive summaries. A count of participants with an event does not convey event severity, timing, recurrence, or causal attribution unless those details are reported. The record supplies counts for serious and other adverse events and deaths, but no further event-level detail in the information. The primary endpoint analysis and the safety summaries answer different questions and should not be treated as interchangeable evidence.

10. Multiplicity, Missing Data and Analysis Populations

The primary endpoint is assessed repeatedly across scheduled time points, and the registry lists multiple secondary outcomes, including concentration relationships, changes in heart rate, PR and QRS intervals, categorical ECG outliers, T-wave morphology and physical examination. In general, testing or estimating many outcomes and time points raises multiplicity concerns: the chance of at least one apparently unusual result can increase when many comparisons are considered. An analysis plan may address this through a prespecified hierarchy, adjustment procedure, or emphasis on estimation rather than multiple significance tests.

The registry record does not report a multiplicity strategy, a hierarchy among outcomes, or which time point or contrast was designated as the principal test. The primary outcome is not reduced to one comparison in the posted results. No adjustment method or family of hypotheses is described, so it is not possible to determine whether any reported inferential results account for multiple outcomes or times. Numerical primary outcome results are not reported in the registry record in any case.

Missing-data methods are also not reported in the registry record. The stated QT/QTc Analysis Set includes participants in the Safety Analysis Set who have baseline measurements and at least one valid post-dose ΔQTcF value. This definition describes eligibility for the analysis set, but does not explain how missing measurements at individual times were handled, whether any values were excluded, or what assumptions were used for incomplete repeated measures. The record does not report the number of participants with endpoint data at each time point or reasons for missing data.

These gaps matter because repeated-measures analyses can be affected by the pattern and reasons for missing observations, and multiplicity decisions affect the interpretation of separate time-specific findings. No assumptions about missingness or adjustment should be attributed to the study without an explicit report. The registry record also does not describe interim analyses, sample-size justification, or a detailed statistical analysis plan. Those details are not reported in the registry record.

11. Safety Results

The registry reports serious adverse events as affected participants divided by participants at risk.

Deaths

Other (non-serious) adverse events

Participants with at least one non-serious event at or above the registry's 0% reporting threshold.

Most frequent non-serious adverse event terms

The safety figures distinguish serious adverse events, other adverse events and deaths. No serious adverse events are reported in the listed groups: 0 among 8 at risk in each Modufolin group and 0 among 9 at risk in the placebo group. Deaths are likewise reported as 0 in each group. The counts for participants affected by other adverse events are 6 in the 200 mg/m2 group, 5 in the 350 mg/m2 group, 8 in the 500 mg/m2 group and 1 in the placebo group.

These are descriptive counts, not estimates of treatment effect. They do not establish whether events were related to study treatment, how severe or long-lasting they were, or whether a participant experienced more than one event. The registry record does not report serious-event terms, and it provides no additional event-level descriptions in the listed safety figures. Zero reported serious events in this study should not be interpreted as proof that serious events cannot occur; the figures describe what was reported for these groups in this record.

The group labels and denominators should be read as recorded. The dose wording elsewhere in the record is inconsistent, and the safety figures do not resolve that discrepancy. These summaries also do not replace the ECG endpoint results, which are not reported in the registry record.

12. What the Result Does, and Does Not, Mean

The record establishes that a Phase I study was conducted to evaluate ECG effects, safety, tolerability and pharmacokinetics after single ascending doses of Modufolin in healthy male volunteers. It identifies change from baseline QTcF as the primary endpoint and describes serial ECG collection before and after dosing. It also reports safety counts by listed dose group and placebo. These registry entries describe the study’s purpose, measurement approach and selected safety summaries.

The numerical primary endpoint findings are not reported in the registry record. As a result, the direction and magnitude of QTcF changes, the differences between dose groups and placebo, and the precision of any estimates cannot be determined. No conclusion about the concentration relationship can be drawn from the listed outcome title alone. The missing results do not mean that the endpoint showed no change; they mean that the registry record does not report the numerical evidence needed to characterize it.

The safety figures report no serious adverse events or deaths in the listed groups and give counts for other adverse events. These observations are descriptive and do not identify cause, severity or comparative risk. They also do not establish the absence of uncommon or later events. The record does not present the detail needed to interpret individual event patterns.

The study’s population is healthy male volunteers, so the stated question is about ECG and safety measurements in that setting. The record does not support generalization to other populations or conclusions about clinical benefit. The description of the study as randomized and placebo-controlled provides design context, but discrepancies in masking, study model and dose wording remain unresolved. The appropriate reading is bounded: the trial evaluated a defined ECG endpoint and reports selected safety counts, while the registry record does not provide the primary outcome estimates required for a quantitative interpretation.

13. Why This Trial Matters Statistically

This trial is statistically notable for its repeated ECG assessment structure. The primary endpoint is measured at multiple scheduled times, with a stated baseline definition based on pre-dose ECG intervals. Serial measurement can be informative because changes may vary over time, but it makes the choice and interpretation of time-specific comparisons important. Without estimates at those times or a stated multiplicity approach, the record cannot show how the outcome evolved or how comparisons were prioritized.

The endpoint is expressed as a model-based least-squares mean with standard error. That choice signals an intended estimate-and-uncertainty framework, but the statistical model and its numerical outputs are not reported. This illustrates a general distinction between knowing an endpoint’s name and measurement plan and having enough results to assess its magnitude and precision. The latter requires estimates and uncertainty information, not design details alone.

The registry also names a concentration-response objective and several ECG secondary outcomes. Together, these indicate that the study addressed more than one statistical question. The record does not describe how those questions were ordered or how multiplicity was handled, so separate findings, if reported elsewhere, would need to be interpreted in light of that missing information.

Finally, inconsistencies in the record’s masking and model descriptions, along with the discrepancy between dose-group titles and the 100 mg reference in an arm description, limit clarity about trial conduct. These do not erase the reported design and safety details, but they make careful attribution important. The main statistical lesson is that a defined serial endpoint and a completed study are not, by themselves, enough to establish the observed ECG pattern; the actual estimates, uncertainty and analysis details are essential.

14. Limitations

• The posted summary statistics are the only data these results rest on; patient-level data, full protocols and statistical analysis plans are not available.
• Recalculated results are simple, unadjusted calculations from posted summaries. They do not replace the sponsor's prespecified analysis.

The most important limitation for interpretation is that numerical results for the primary endpoint are not reported in the registry record. Although ΔQTcF is identified as the primary outcome and its measurement schedule is described, no least-squares mean values, standard errors, confidence intervals or p-values are available in the record. The magnitude, direction and precision of the ECG findings therefore cannot be assessed.

The record does not report a detailed statistical analysis plan, including the model specification, handling of repeated measures, prespecified primary time point, contrast definitions or multiplicity strategy. It also does not report missing-data assumptions, the number with valid measurements at each time point, or reasons for missing observations. These omissions prevent reconstruction of the primary analysis and limit evaluation of how the reported analysis set was applied.

There are design-description inconsistencies. The record labels masking as triple, while the official title says double-blind. The model field says single-group even though the listed group descriptions include three Modufolin doses and placebo. The dose-group titles state 200 mg/m2, 350 mg/m2 and 500 mg/m2, while the Modufolin arm description refers to 100 mg. The registry record does not explain these discrepancies, so the precise design and dose wording cannot be resolved from the record alone.

Safety summaries are limited to counts of serious and other adverse events and deaths. The record does not include event-level descriptions, timing, severity or causal assessment in the listed safety information. The population is described as healthy male volunteers, which defines the setting but does not establish applicability to other populations. Details of sample-size justification, adaptive rules and interim analyses are not reported in the registry record.

15. Related Tutorials

16. Related Statistical Calculators

17. Sources

• ClinicalTrials.gov: Study Evaluating ECG Effects, Safety, Tolerability and Pharmacokinetics of Single Doses of Modufolin (Arfolitixorin) in Healthy Volunteers Tetrahydrofolate in Healthy Male Volunteers — NCT03203564.
• Registry record last updated 2020-09-25; status last verified 2020-09.
• Results first posted 2020-03-19.
• Data retrieved from ClinicalTrials.gov on 2026-11-09.
• Group sizes, means, event counts and sponsor-reported p-values are reproduced as posted. Effect estimates, confidence intervals and p-values not attributed to the sponsor are independently calculated from the posted summary statistics.

18. Record Summary

The registry record describes NCT03203564 as a completed Phase I study sponsored by Isofol Medical AB, with enrollment of 33. It concerns single ascending doses of Modufolin, also called arfolitixorin, in healthy male volunteers and lists placebo as the comparator. The study start is recorded as 2017-07-04 and primary completion as 2017-08-25.

The primary endpoint is change from baseline QTcF in msec, with least-squares mean and standard error identified as the parameter and dispersion. ECG assessments were scheduled before and after dosing, with baseline defined from measurements at 45, 30 and 15 min before dosing. The registry also lists secondary ECG, safety, pharmacokinetic and physical-examination outcomes.

The safety figures report no serious adverse events or deaths in any listed group and give counts for other adverse events. Numerical results for the primary QTcF endpoint and the concentration relationship are not reported in the registry record. The record also contains unresolved inconsistencies in masking, study model and dose wording.