HCQ on PAR-4 Levels in Patients With Resectable Solid Tumors: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for HCQ on PAR-4 Levels in Patients With Resectable Solid Tumors (NCT02232243), a non-randomized phase 1 study. The primary outcome is Patients With Elevated Par-4 Levels.

2. Clinical Question

This is a biological-response question: it concerns a measured change in a plasma marker during a short pre-surgical course of HCQ. It is not, as described in the registry, a comparison of clinical outcomes between HCQ and no HCQ. Nor does the primary endpoint ask whether tumors shrank, whether surgery was more successful, or whether patients lived longer.

The record’s background rationale is that normal cells generally secrete PAR-4 at levels considered inadequate to cause substantial apoptosis in cancer cells, and that HCQ’s apoptosis-sensitizing features may enhance effects of cancer therapies. These are the investigators’ rationale and hypothesis, not findings established by this trial. The posted registry record does not report how many participants met the endpoint, the measured PAR-4 values, or an estimate of the change. Accordingly, the clinical question can be described from the design, but its answer cannot be determined from the endpoint results in the registry record.

3. Trial Design

The registry lists this as a phase 1 pilot study with a parallel model, non-randomized allocation and no masking. It reports enrollment of 10 participants and a completed status. Participants had resectable solid tumors and received HCQ before surgery. The course lasted 14 days in each described group.

The sequence began with three participants taking HCQ 200 mg twice daily, described as 400 mg/day total. Based on data analysis from these initial patients, the secondary group received HCQ 400 mg twice daily, described as 800 mg/day total. Based on data from the primary and secondary groups, the tertiary group received HCQ 200 mg twice daily, again described as 400 mg/day total. These are sequential cohorts, not randomized dose arms. The dose used in the secondary group differed from the dose used in the initial and tertiary groups, and the timing of enrollment was tied to prior group data.

That structure matters when interpreting any apparent differences between groups. Without random assignment, group membership may be connected to the order in which participants entered the study, as well as to dose. The registry does not describe a concurrent untreated or standard-care control group, and it cautions that the arms are not a like-for-like control comparison. The primary endpoint instead refers to comparing each participant’s baseline and day 14 PAR-4 levels.

The registry record does not report the rationale for the group sizes, the criteria used to move between groups, or the details of the analysis used to inform those decisions. Further protocol information about eligibility, recruitment and study conduct is not reported in the registry record. The stated time point for the primary endpoint is baseline and day 14; the record does not explain how measurements were collected or handled if a participant did not reach that time point.

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: 10 patients enrolled in this dose escalation study. In Period 1, three patients were assigned to receive 200mg HCQ twice daily. Based on toxicity and PAR-4 levels, in Period 2 the next four patients received 400mg HCQ twice daily. Following additional analysis, in Period 3 the final three patients received 200mg HCQ twice daily.

First Dose (200mg HCQ Twice Daily)

Second Dose (400mg HCQ Twice Daily)

Reasons for not completing this period:

Third Dose(200mg HCQ Twice Daily)

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 Patients With Elevated Par-4 Levels, measured over Baseline and day 14.

The primary endpoint is the number of patients with a 2-fold change in PAR-4 levels from baseline to day 14. The registry gives the unit as participants and describes the timeframe as baseline and day 14. The brief summary expresses the study hypothesis as an increase of at least 2-fold in systemic, plasma PAR-4 levels compared with pretreatment plasma levels.

This endpoint turns a continuous biomarker measurement into a participant count: each patient either meets the stated change criterion or does not. A count can be easy to interpret as the number of participants reaching a prespecified biological threshold. It also discards information about the size and direction of changes beyond that threshold. For example, the count alone would not describe the underlying PAR-4 values or how far participants’ changes were from the cutoff.

In general, a biomarker endpoint can be used to examine whether an intervention is associated with a biological response. It is not automatically evidence of a clinical benefit. For this trial, the registry does not report the number meeting the endpoint, individual or group-level biomarker values, or a statistical estimate of change. The endpoint’s observed result is therefore not reported in the registry record. The registry record also does not give details of the second outcome measure beyond listing that the study has two outcome measures.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

What kind of outcome is the primary endpoint?

The endpoint is binary at the participant level: the registry counts patients with a 2-fold change in PAR-4 levels from baseline to day 14. A binary endpoint records whether a specified condition was met, rather than the full numerical value of a measurement. The reported unit is participants, so the intended summary is a count of people meeting the criterion. The registry does not report that count or the underlying measurements.

In general, converting a continuous biomarker into a threshold-based outcome can make a result straightforward to state, but it loses detail. The count cannot show the distribution of changes, how close participants were to the cutoff, or whether values above the cutoff were only just above it or substantially greater. Reporting the biomarker measurements alongside the threshold count would usually provide complementary information, but those measurements are not reported in the registry record.

Why does the baseline-to-day-14 structure matter?

The endpoint refers to two time points for participants: baseline and day 14. In general, measurements from the same participant are paired, because the two observations share the participant’s characteristics. A statistical analysis of change should account for that pairing rather than treating the two measurements as if they came from unrelated people. The record does not describe the statistical method used for the endpoint, and it reports no analysis results.

The threshold also needs consistent operational definition to support interpretation. The registry describes a 2-fold change and the brief summary specifies a hypothesized increase of at least 2-fold, but does not give further details about data processing, assay procedures, or how measurements at or near the threshold were handled. Those details are not reported in the registry record.

What do allocation and masking imply?

Allocation was non-randomized, and masking was listed as none. In general, randomization helps balance participant characteristics across comparison groups, on average, while masking can reduce some forms of bias in assessment or care. Neither feature is described for this trial. The three groups were sequential, with dose changes informed by data analysis from earlier participants. Consequently, group contrasts could reflect dose, enrollment order or other differences, and cannot be interpreted as randomized comparisons.

The registry does not describe a concurrent control group. Its primary question is framed around change from baseline to day 14, which is a before-and-after comparison within participants. Such a comparison can document change over time, but, in general, it cannot by itself distinguish an intervention-associated change from other influences occurring over the same period.

What can be said about uncertainty and testing?

A confidence interval is a statistical range expressing the uncertainty around an estimate under a specified method. In general, wider intervals indicate less precision, while narrower intervals indicate greater precision. The registry does not report a confidence interval or another uncertainty estimate for the primary endpoint. It also does not report a p-value or the statistical test used. Without an estimate and its uncertainty, the size and precision of the biomarker response cannot be evaluated from this record.

The record lists zero analyses. This does not establish that no analysis was ever conducted; it means statistical analysis results are not reported in the registry record. The registry also does not give a statistical analysis plan, a sample-size justification, or details of any interim analyses. Those matters should not be inferred from the fact that group doses were changed after earlier patients.

How should planning calculations be read?

General sample-size planning estimates describe how many participants might be needed under stated assumptions to detect an effect with a specified design. The planning illustration associated with this study uses a two-sided significance level of 0.05, equal allocation between two arms and a normal approximation, with the effect framed as a fraction of the observed effect. These are planning assumptions, not reported features of the trial’s actual non-randomized sequence.

Any such sample-size illustration is not observed power and does not show what the trial achieved. It also does not establish that the original study was designed or justified using those assumptions. The registry does not report its own sample-size justification, and numerical planning results are not reported in the registry record. In general, power is a design concept based on assumptions about an effect and variability; it is not a retrospective measure of whether a study result is reliable.

Over-dispersion is a general concern when count data vary more than a simple statistical model expects. It can make standard errors too small if the model does not allow for extra variability. The primary endpoint is a participant count meeting a binary criterion, but the registry does not describe a model for that endpoint, so whether over-dispersion was assessed is not reported in the registry record.

10. Exact Estimates for Each Arm

No two-arm comparison is shown for this trial, but each arm's result can still be summarized with exact interval estimates computed from the posted counts. The Clopper-Pearson interval is exact and conservative; the Wilson interval is a common closer-to-nominal alternative; the Bayesian credible interval uses a uniform Beta(1, 1) prior. For small groups the three can differ noticeably.

R code and output

arm    <- c("Initial: Hydroxychloroquine 400mg HCQ", "Secondary: Hydroxychloroquine 800mg HCQ", "Tertiary: Hydroxychloroquine 400mg HCQ")
n      <- c(3, 3, 3)
events <- c(3, 2, 1)
for (i in seq_along(n)) {
  ci <- binom.test(events[i], n[i])$conf.int          # Clopper-Pearson exact interval
  cr <- qbeta(c(0.025, 0.975), 1 + events[i], 1 + n[i] - events[i])   # Bayesian, uniform Beta(1,1) prior
  wi <- (function(x, n) { p <- x / n; z <- qnorm(0.975); d <- 1 + z^2 / n
    c((p + z^2 / (2 * n) - z * sqrt(p * (1 - p) / n + z^2 / (4 * n^2))) / d, (p + z^2 / (2 * n) + z * sqrt(p * (1 - p) / n + z^2 / (4 * n^2))) / d) })(events[i], n[i])
  cat(sprintf("ARM|%s|%d|%d|%.1f|%.1f|%.1f|%.1f|%.1f|%.1f|%.1f\n", arm[i], n[i], events[i], 100 * events[i] / n[i],
              100 * ci[1], 100 * ci[2], 100 * wi[1], 100 * wi[2], 100 * cr[1], 100 * cr[2]))
}

ARM  Initial: Hydroxychloroquine 400mg HCQ  3  3  100.0  29.2  100.0  43.9  100.0  39.8  99.4
ARM  Secondary: Hydroxychloroquine 800mg HCQ  3  2  66.7  9.4  99.2  20.8  93.9  19.4  93.2
ARM  Tertiary: Hydroxychloroquine 400mg HCQ  3  1  33.3  0.8  90.6  6.1  79.2  6.8  80.6

11. Multiplicity, Missing Data and Analysis Populations

The registry lists two outcome measures, but gives detailed information in this record only for the primary measure: the number of patients with a 2-fold change in PAR-4 levels from baseline to day 14. The details and results for the other outcome measure are not reported in the registry record. It also lists zero analyses, and does not report a statistical analysis plan, the tests used, or analysis results.

Multiplicity arises in general when a study makes several statistical comparisons or tests several outcomes. Testing more hypotheses can increase the chance of finding at least one apparently noteworthy result by chance, so a study’s analysis plan may specify how such testing is handled. The registry does not report whether multiplicity adjustments were planned or used, nor does it describe a hierarchy among outcomes beyond identifying the primary endpoint. It is therefore not possible to assess the handling of multiple outcomes or comparisons from the record.

The registry does not report missing-data counts or describe how missing baseline or day 14 measurements were handled. It does not state whether participants without a day 14 result were excluded, counted as not meeting the threshold, or handled by another approach. Those choices can affect an endpoint based on change between two time points. No imputation approach or sensitivity analysis is reported in the registry record.

The record also does not define analysis populations, such as whether analyses included all enrolled participants or only participants with both measurements. Although the registry reports enrollment of 10, it does not give a participant flow or explain how many contributed to each endpoint analysis. The resulting uncertainty about missingness, analysis populations and multiplicity further limits interpretation of any unreported primary result.

12. 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 registry lists serious adverse event counts by group. It reports 0 serious adverse events among 3 participants in the initial group, 0 among 4 participants in the secondary group, and 0 among 3 participants in the tertiary group. No serious adverse event terms are listed. These are descriptive counts for the groups as recorded; they do not establish that serious events could not occur or that HCQ caused no serious events.

Other adverse events are listed separately: 1 among 3 participants in the initial group, 2 among 4 in the secondary group, and 1 among 3 in the tertiary group. The registry also lists deaths of 1, 0 and 1, respectively, for those groups. The record does not report the event descriptions, timing, causes of death, or whether any event was considered related to HCQ. The death counts should therefore be read as registry entries, not as evidence of treatment causation.

Because groups were sequential and not randomized, these counts do not support a direct comparative safety conclusion between doses. In general, event counts in small groups are descriptive and may be imprecise. The registry does not report a statistical comparison or additional context that would clarify the events.

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

The study addresses a focused biological question: whether PAR-4 levels in plasma changed from baseline to day 14 during a pre-surgical course of HCQ, with a stated threshold of 2-fold. It also records adverse events and deaths across three sequential dose groups. The registry describes a phase 1 pilot with enrollment of 10 participants, but it does not report the primary endpoint result.

That missing result is central. Without the count of participants meeting the endpoint, measured PAR-4 values, or an uncertainty estimate, the record cannot establish whether the hypothesized biomarker change was observed or how precisely it was estimated. The investigators’ stated rationale and hypothesis explain why the question was asked, but do not answer it.

Even if a biomarker change were reported, it would not alone demonstrate a clinical benefit. PAR-4 is the biological measure identified in the primary endpoint; the endpoint is not a measure of tumor response or another clinical outcome. The registry record does not report such outcomes or establish that any biomarker change would translate into a patient benefit.

The study’s design also does not support a clean randomized dose comparison. The groups were enrolled sequentially, and the secondary group received a different dose from the initial and tertiary groups. The registry notes that the arms are not a like-for-like control comparison. Accordingly, any observed difference across groups would need cautious interpretation and could not be attributed to dose alone on the basis of this design.

The adverse-event counts offer a limited description of events recorded in the groups, but the absence of serious adverse events in the registry entries does not prove absence of risk. The record does not provide enough detail to assess causality, event timing, or comparative safety. Overall, the registry outlines the study question, exposure schedule and some safety counts, while leaving the main biomarker result and important analysis details unreported.

14. Why This Trial Matters Statistically

This trial is statistically notable because its main endpoint is a threshold-based biomarker outcome measured over time within participants, while the dose groups were introduced sequentially rather than randomly. Those features shape what the design can answer. A baseline-to-day-14 comparison can describe within-participant change, but without a concurrent control it cannot separate the intervention’s contribution from other changes over the same period.

The sequence of dose groups adds another interpretive constraint. The initial and tertiary groups received the same dose, while the secondary group received a higher dose, and each later group was informed by data from earlier participants. This may be relevant to a pilot study’s exploratory purpose, but it does not create a randomized dose comparison. The registry cautions against treating the arms as like-for-like controls.

The endpoint’s binary threshold is another important statistical choice. Counting participants who cross a stated cutoff can produce an accessible summary, but it omits information about the magnitude and distribution of the underlying biomarker changes. The registry does not report the endpoint count or measurements, so neither the threshold summary nor the fuller biomarker pattern can be assessed.

Finally, the case illustrates why an enrollment figure and a completed status do not substitute for analysis results. The registry reports enrollment of 10 and a completed status, yet does not post the primary endpoint result, uncertainty estimates or analysis methods. Without those details, the study’s precision and findings cannot be evaluated. The design and endpoint can be described, but the central statistical question remains unanswered in the registry record.

15. 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 record reports a small pilot enrollment of 10 participants and provides no primary endpoint result. This leaves the principal biological question unresolved in the posted registry information. No PAR-4 values, count meeting the 2-fold threshold, confidence interval, p-value or other analysis result is reported.

The non-randomized, sequential design is a specific limitation for interpreting differences by dose. The groups entered at different stages, and changes in dose followed data analysis from earlier groups. There is no like-for-like randomized comparison described. The record does not report a concurrent control group, so a baseline-to-day-14 change would not by itself establish what would have happened over the same period without HCQ.

The endpoint reports a threshold count rather than the full biomarker distribution. The registry does not describe assay methods, data handling, or the exact operational treatment of measurements around the threshold. It also does not give details for the second listed outcome measure. The statistical analysis plan, sample-size justification, interim-analysis procedures, and analysis populations are not reported in the registry record.

Missing-data information is also absent: the record does not state whether all enrolled participants had both measurements or how unavailable measurements were handled. Safety counts are listed, but event terms, timing, causes, attribution and comparisons are not given. The registry’s serious-event counts and deaths therefore provide limited context and cannot support conclusions about causality. Together, these omissions restrict assessment of effect size, precision, safety and the robustness of any possible result.

16. Related Tutorials

17. Related Statistical Calculators

18. Sources

• ClinicalTrials.gov: HCQ on PAR-4 Levels in Patients With Resectable Solid Tumors — NCT02232243.
• Registry record last updated 2021-06-28; status last verified 2019-05.
• Results first posted 2019-07-23.
• 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.

19. Record Summary

The registry describes a completed phase 1 pilot study of HCQ in patients with resectable solid tumors, with enrollment of 10. The primary question concerns the number of patients with a 2-fold change in PAR-4 levels from baseline to day 14. Participants received HCQ twice daily for 14 days before surgery in three sequential groups: the initial and tertiary groups received 200 mg twice daily, and the secondary group received 400 mg twice daily.

Allocation was non-randomized and masking was absent. The registry cautions that the groups are not a like-for-like control comparison. It lists serious and other adverse-event counts and deaths by group, but does not report event descriptions or attribution. The primary endpoint result, statistical analyses, uncertainty estimates, missing-data handling and analysis populations are not reported in the registry record. Thus, the record describes the study question and design but does not provide the result needed to assess its main biomarker endpoint.