Efficacy and safety of moxidectin and albendazole compared to ivermectin and albendazole co-administration in adolescents infected with Trichuris trichiura: a randomized controlled trial protocol

Research objectives

We designed a randomized controlled trial to show non-inferiority of co-administered moxidectin/albendazole compared to co-administered ivermectin/albendazole in participants aged 12–19 years on Pemba Island, Tanzania. From evidence of previous studies conducted by our research group, we expect similar efficacies from the moxidectin/albendazole combination compared to ivermectin/albendazole^8,9. Nevertheless, moxidectin might be advantageous in terms of the drug’s longer half-life and as an alternative in areas with possible emerging ivermectin resistance^16,18. As recommended for new combination therapies, we simultaneously assess superiority of the drug combinations compared to monotherapies.

The primary objective is to demonstrate non-inferiority of co-administered moxidectin (8 mg)/albendazole (400 mg) compared to combined ivermectin (200 µg/kg)/albendazole (400 mg) in terms of egg reduction rates (ERRs) against T. trichiura infections assessed by Kato-Katz at 14–21 days post-treatment in adolescents aged 12–19 years with a non-inferiority margin of 2 percentage-points and 90 power at the 95% confidence interval.

The secondary objectives of the trial are:

This article is reported in line with the Standard Protocol Items: Recommendations for Interventional Trials (SPIRIT) guidelines¹⁹.

Trial design

A phase 3 randomized controlled open-label non-inferiority trial with blinded outcome assessors will be conducted among adolescents aged 12–19 years with T. trichiura infection. The trial includes one baseline and three follow-up assessments at 14–21 days, 5–6 weeks and 3 months post-treatment (Figure 1). The study is designed as a five-arm trial including two arms with combined treatment through co-administration of separate tablets (arm A; moxidectin/albendazole, arm B; ivermectin/albendazole) and three arms with single drug administration (arm C; albendazole, arm D; ivermectin, arm E; moxidectin).

The efficacy of the treatments will be determined by collecting two stool samples before and at every post-treatment time-point. Each sample will be microscopically examined for T. trichiura eggs using duplicate Kato-Katz thick smears. Participants will be eligible if they are positive (≥ 48 eggs per gram of stool (EPG)) for T. trichiura eggs at baseline and will be considered cured at the different follow-up assessments if no T. trichiura eggs are found in the stool samples.

Figure 1. Design and timeline of the randomized controlled trial to be implemented on Pemba Island (Pemba Island, Tanzania).

The study is designed as a five-arm trial including two arms with combined treatment through co-administration of separate tablets (arm A; moxidectin/albendazole, arm B; ivermectin/albendazole) and three arms with single drug administration (arm C; albendazole, arm D; ivermectin, arm E; moxidectin). Abbreviations: ALB, albendazole; MOX, moxidectin; IVM, ivermectin; PK, pharmacokinetics; AE, adverse events.

Outcomes

Primary outcome. The primary outcome is the ERR of T. trichiura egg counts assessed by Kato-Katz microscopy pre-treatment and 14-21 days post-treatment.

Secondary outcomes. The secondary outcomes include CR, defined as conversion from being egg positive pre-treatment to egg negative post-treatment, of T. trichiura as well as CRs and ERRs for A. lumbricoides and hookworm assessed by Kato-Katz at 14–21 days post-treatment. In addition, tolerability of treatment (type, number and severity of adverse events (AEs)), infection status and intensity derived by Kato-Katz at 5–6 weeks and 3 months post-treatment and PK/PD parameters of the study drugs will be assessed.

Study area and participants

This trial will be implemented as a school-based study on Pemba Island (Zanzibar, on Pemba Island, Tanzania). Secondary schools in areas with moderate to high T. trichiura prevalence will be selected based on experience from earlier studies and/or knowledge of the local collaborating teams. These will be Kilindi, Kwale and Ndagoni located in Chake Chake district as well as Kiuyu in Wete district. In each selected school, adolescents aged 12–19 years will be invited for study participation. Entering school over-age is a common occurrence in Zanzibar, thus ages of secondary level pupils may well range from 12 to 19 years of age. Adolescents are within the main target group of helminth control programs and are listed among potential receivers of moxidectin that is, so far, only approved from 12 years of age onwards²⁰.

Study duration

Screening for the baseline is scheduled to start 3 months prior to treatment. Follow-up screenings will take place between 14–21 days, 5–6 weeks and 3 months post-treatment and will last approximately two weeks, respectively. Thus, the maximum time for subject participation will be 6 months. The schedule of visits are summarized in Table 1.

Recruitment

School teachers and caregivers of potential participants and adolescents aged ≥18 years will be invited to participate in an information session. The research team will explain the purpose and procedures of the study, as well as potential benefits and risks of participation. Attendees will be encouraged to ask questions which will be discussed in an open setting. Caregivers interested in having their child/children of 12–17 years of age participate in the study or adolescents aged 18–19 years willing to participate will be invited to complete the process of informed consent by signing the informed consent form (ICF)¹⁹. In addition, written assent will be obtained from underage participants. Participants having a signed ICF will be assessed for eligibility.

Laboratory procedures

At baseline, all participants will be asked to provide two stool samples (within a maximum of seven days). From each stool specimen, duplicate Kato-Katz thick smears (41.7 mg each)²¹ will be prepared and examined under a microscope for eggs of T. trichiura, A. lumbricoides and hookworm by skilled technicians.

For quality control of T. trichiura and A. lumbricoides egg counts, 10% of slides will be re-read by another laboratory technician. To ensure quality of hookworm diagnosis, 10% of the stool samples will be divided into two sub-samples; one of the containers will keep its original participant ID, whereas the second container will be labeled with a new ID (assigned by the co-PI). An additional Kato-Katz slide will be made from the second container and the findings compared to the ones from the original sample. For hookworm, results are considered correct if no difference in presence/absence of helminth eggs is found. For T. trichiura and A. lumbricoides the following tolerance margin should not be exceeded: (i) no difference in presence/absence of T. trichiura and A. lumbricoides (ii) egg counts are +/-10 eggs for counts ≤100 eggs or +/-20% for counts >100 eggs (for each species separately). In case discrepancies above the tolerance margin are noted, the respective slides are examined a third time. The new results are discussed to reach consensus. The same sampling procedure, diagnostic approach and quality control assessment will be applied at 14–21 days, 5–6 weeks and 3 months post-treatment.

Eligibility criteria

Participants meeting all inclusion criteria and none of the exclusion criteria (Table 2) will be invited for treatment.

Clinical assessment

A clinical examination of the study participants assessing general health, anthropometric parameters including height and weight as well as forehead temperature using a Braun No touch + forehead NTF3000 (Braun GmbH, Kronberg, Germany) thermometer will precede the treatment. Each participant will be asked to provide a finger-prick blood sample for hemoglobin (Hb) levels, which will be measured using a HemoCue analyzer (Hb 301 system, Angelholm, Sweden). To avoid accidental treatment of pregnant girls/women all female participants will be asked to provide a urine sample for a pregnancy rapid diagnostic test at baseline and at the end of the study (3 months after treatment). Girls/women will be individually counselled that they should not become pregnant during the entire study period. All trial participants will further be asked about chronic diseases and existing clinical symptoms the day of exam, which will be evaluated on relevance with regard to exclusion criteria (Table 2).

Criteria for discontinuation of trial

A subject can be discontinued from the study for the following reasons:

Discontinued subjects will not be replaced. If, for any reason, a subject is discontinued from the study after treatment but before the end of treatment evaluations, the safety assessment will still be conducted to ensure the discontinued participant’s well-being. Data obtained prior to the withdrawal will be included in the analysis to ensure the validity of the trial.

Randomization, concealment and masking

Study participants eligible for treatment will be randomly assigned to one of the five treatment arms using a computer-generated stratified randomization code. The random allocation sequence will be generated by using an algorithm which minimizes deviations from the anticipated arm sizes stratified by 2 levels of baseline infection intensity (light: 1-999 EPG, and moderate/heavy plus heavy: ≥ 1000 EPG T. trichiura infections), which will be provided by the trial statistician not involved in enrolment, treatment and data collection. This ensures that all treatment arms will have a similar proportion of participants with light infection intensity. The number of light versus moderate/heavy infections, however, are not expected to be equal in each arm, depending on the distribution of infection intensity in the recruited cohort. Team members conducting the treatment will not know the allocation order. Concealment will be warranted by masking the randomization sequence using envelopes containing the respective treatment arm labels. The study is defined as open-label yet masking is assured since the primary outcome assessors, i.e., the microscopists determining the egg counts for the efficacy assessment, will have no knowledge of the participants’ assignment to treatment arms.

Treatment

All eligible T. trichiura-infected participants will be treated with the respective single or combination treatment regimen at day zero. All regimens are administered orally as a single dose. 400 mg albendazole will be the product of Glaxo Smith Kline (Zentel®) and a single tablet administered. 3 mg tablets of ivermectin will be obtained from Merck (Stromectol®). The weight will be recorded for each participant and the correct dose (i.e., 200 µg/kg) evaluated and administered accordingly. Moxidectin 2 mg tablets will be obtained from Medicines Development for Global Health and 4 tablets administered to each participant. All drugs will be given in the presence of the investigator(s), and ingestion confirmed. This will be recorded with the time and date of administration. Subjects will be asked not to take any drugs other than those prescribed by the study medical team. After ingestion of the medication, the subjects will be observed for 3 hours to ensure retention of the drug. Vomiting and spitting within 1-hour post-dosing will require re-dosing. The subjects will not be allowed more than one repeated dose. No re-administration will be needed for subjects vomiting after one hour. The PI and/or Co-PI is responsible for drug accountability at the study site. Maintaining drug accountability includes careful and systematic study drug storage, handling, dispensing and documentation of administration.

At the end of the study all participants remaining positive for any STH infection will be treated with the currently best recommended treatment (i.e., ivermectin/albendazole against T. trichiura and hookworm and albendazole against A. lumbricoides).

Pharmacokinetic studies

The PK study will be performed in a maximum of 15 participants in the combination chemotherapy treatment arms (i.e., arms A and B) and 10 participants in the monotherapy treatment arms (i.e., arms C-E), amounting to a subsample of 60 participants overall. All study participants, regardless of participation in PK, will receive a local high-fat breakfast before treatment¹⁷. Since population PK parameters of all three study drugs are available^23–25, a sparse sampling approach will be applied to describe the population-based PK profiles of the individual drugs upon mono- or co-administration. Additionally, potential interference between moxidectin or ivermectin and albendazole will be assessed. For this, capillary blood (≤60 µL) will be collected by puncture with a finger prick at four time points (approx. 6h, 21h, 27h, 45h post treatment). Two microsamples (duplicates) will be taken at each time point. Each time, the drop of blood will be directly transferred onto Mitra® sticks (Neoteryx, Toronto CA) (10 µL or 30 µL) and/or on Whatman® protein saver cards 903 filter paper (Merck, Darmstadt DE) (30 µL). Mitra® sticks will be utilized for participants having received albendazole and/or moxidectin, whereas filter paper will be used for ivermectin-treated participants. The dried Mitra® sticks and filter paper will be transported to Swiss TPH, Basel, and stored at room temperature until analysis within one month after blood collection. The quantification of the study drugs will be performed using validated liquid chromatography tandem mass spectrometry (LC-MS/MS) methods as described elsewhere^23–25. Drug concentrations will be calculated by interpolation from a calibration curve with a lower limit of quantification of 1–5 ng/ml. 7% of the sample duplicates will be analyzed for quality control, and the measured concentrations will be used to determine between-run and overall precision and accuracy of the analysis.

Safety assessments

Few adverse events (AEs) have been reported following albendazole, ivermectin or moxidectin single and co-administration in STH-infected individuals. The most common AEs were abdominal cramps, headache, itching, fatigue, nausea, diarrhea, fever and vertigo^{7–9,16,26–28}.

Interviews will be conducted to determine the emergence of clinical symptoms directly before treatment within the scope of baseline assessment. Participants will be kept for 3 hours after treatment administration to observe any possible acute AEs and reassessment will be done at 24 hours post-treatment. The local study physician will perform a full clinical examination if moderate to severe and/or unexpected AEs occur, and findings will be recorded. An emergency kit will be available on site to treat any medical conditions that warrant urgent medical intervention. At 3 and 24 hours after treatment and retrospectively at days 14–21 as well as 5–6 weeks and 3 months post-treatment, participants will again be interviewed for the assessment of AEs. Information on all AEs (incidence, intensity, seriousness and causality) will be entered immediately in the appropriate AE module of the case report form (CRF). For all AEs, sufficient information will be pursued and/or obtained so as to permit i) an adequate determination of the outcome of the event (i.e., whether the event should be classified as a serious adverse event (SAE)); and; ii) an assessment of the causal relationship between the AE and the study treatments. Intensity of AE will be judged by the study physician (active assessment) or a trained team member (retrospective assessment), following guidelines by the European Medicine Agency (ICH E2A Clinical safety data management: definitions and standards for expedited reporting)²⁹. Serious adverse events that are still ongoing at the end of the study period will be followed up to determine the final outcome. Any study-related unanticipated problem posing risk of harm to subjects or others (including all unexpected adverse drug reactions), and any type of SAE will be immediately (within a maximum of 24 hours after becoming aware of the event) notified to the study Sponsor-Investigator and co-PIs. Symptoms arising within the time span of 24 hours after treatment and the respective follow-up time points will be monitored passively by teachers or local health workers who will report incidences to the study team. All pregnancies will be reported to the Sponsor-Investigator promptly after becoming aware of the pregnancy. A study physician recruited from a local health facility/hospital will serve as medical contact between the study team and the treating physician or take up the role of treating physician directly. The treating physician will follow-up on the study participant until the end of the pregnancy (either by birth or resolved otherwise). The outcome of the pregnancy will be reported to the Sponsor-Investigator.

Data management and data quality control

Investigators of Swiss TPH and Public Health Laboratory - Ivo de Carneri (PHL-IdC) have agreed on the protocol, performance of study procedures (SOPs from previous studies available on site), CRF completion, specimen collection and diagnostic methods prior to the initiation of the study.

CRF data will be double-entered and compared using Beyond Compare 4 (Scooter Software Inc., Madison, Wisconsin). Any discrepancies will be reviewed against the hard copies of the CRF and corrected accordingly. Electronic data files will be stored on secured network drives with restricted access to study personnel only. Data analysis will be conducted with pseudonymized data and reporting of findings will be fully anonymized.

Source data. Source data are comprised of clinical findings and observations as well as laboratory data maintained and compiled at the study site. Source data are contained in source documents and are allowed to be accessed by local authorities. Source data will be directly entered in the following documents:

Data collection and documentation. Data collected and produced within this trial will fall into one of the following categories:

Data for categories a) to c) will be recorded both paper-based and directly into tablets using CommCare (Dimagi, Inc., Cambridge, MA) or computers, whereas data in category d) will be captured by software only. Data compiled using the software will be directly saved on the personal, password-protected laptop of one of the Co-PIs and uploaded to a server hosted at Swiss TPH, Basel. In paper-based data collection, all missing data must be explained. If an item on the CRF is left blank because the procedure was not done or the question was not asked “N/D” will be entered. If the item is not applicable to the individual case “N/A” will be written. All entries will be printed in black ink. All corrections must be noted with the initials of the respective team member and dated. Data in categories a) and b) will be merged into a masterfile and saved in .xlsx, .mdb and/or .csv. Paper-based data will serve as a physical backup and the source data. Data in categories c) and d) will be saved as .mdb, .csv, .xlsx, .txt and/or .pdf files.

Data storage and preservation. All samples will be destroyed after completion of the study. Paper-based and/or electronic source data and related material will be preserved for a minimum of 15 years to enable understanding of the study procedures, which allows the work to be assessed retrospectively and repeated if necessary. The study site will retain a copy of the documents to ensure that local collaborators can provide access to the source documents to a monitor, auditor, or regulatory agency. Electronic source documents will be stored on a flash drive and kept at the study site (IdC PHL, Pemba Island, Tanzania). The primary data storage and backup will be in the Swiss TPH shared server and secondary data storage will be on personal, password-protected laptops. Electronic data files and archiving conditions will be made strictly confidential by password protection.

Ethical, legal and confidentiality issues. Information about study subjects will be kept confidential and managed accordingly. Screened participants will be listed in a confidential “subject screening log” and attributed a unique study ID. In case of enrolment, participants will be listed in a confidential “subject enrolment log”; this document will constitute the only source to decode the pseudonymized data and will only be accessible to the investigators. Personal data will be coded for data analysis. No names will be published at any time, and published reports will not allow for identification of single subjects. Confidentiality will be ensured throughout the entire research project. All databases will be password secured. None of the investigators declare to have any conflicts of interest.

Statistics

Sample size calculation. For the primary analysis the trial is designed as two arm parallel group randomized controlled trial. We test the primary hypothesis that the treatment combination moxidectin and albendazole is not inferior compared to ivermectin and albendazole. To determine the required sample size, we run a series of simulations using artificial data which behaved roughly in the same way as found by Barda et al.⁸. Assuming true ERR of 98% in both arms, we estimate that 160 participants are required in each group to be at least 90% sure that the limits of a two-sided 95% confidence interval (CI) will exclude a difference in favor of the standard group of more than 2 percentage points. To account for a potential loss to follow-up of 10% and including a safety margin of 20% to account for uncertainty in our assumptions underlying the simulations, we anticipate enrolling 210 participants in each combination treatment arm (arm A and B). The secondary hypothesis anticipates superiority of combination therapies against monotherapies. Assuming CRs below 25% for albendazole as well as for ivermectin and 40% for moxidectin monotherapy, we need to enroll 20, 20 and 80 adolescents, respectively, to identify a statistical significant difference with 85% to 90% power (arm C, D, E)^{1,6,9,30–32}. We thus aim to recruit 210 + 210 + 20 + 20 + 80 = 540 participants in total.

The suggested sample size of a maximum of 4 PK time points from 60 willing participants (10–15 per study arm) is sufficiently high to determine the population PK parameters and investigate potential drug-drug interactions with a sparse sampling scheme, considering that PK variability is moderate. A moderate PK variability is a reasonable assumption when dealing with adolescents.

Description of statistical methods. In non-inferiority trials, non-inferiority has to be demonstrated in the intention-to-treat and in the per protocol population. The primary analysis will be performed according to the intention-to-treat principles using the available case population, which includes all participants with any primary end point data. Subsequently, a per-protocol analysis will be performed. Eggs per gram of stool will be assessed by calculating the mean egg count from the quadruplicate Kato-Katz thick smears and multiplying this number by a factor of 24. The geometric mean (GM) ERR will be calculated as:

GM egg counts will be calculated for the different treatment arms before and at 14–21 days after treatment to assess the corresponding ERRs. Bootstrap resampling method with 5,000 replicates will be used to calculate 95% CIs for ERRs and the difference between the ERRs.

CRs will be calculated as the percentage of egg-positive adolescents at baseline who become egg-negative after treatment. Differences among CRs will be assessed by using unadjusted logistic regressions. In a subsequent analysis an adjusted logistic regression (adjustment for baseline infection intensity, age, sex, weight) will be performed. ERRs and CRs as well as their respective confidence intervals will be calculated separately for each follow-up. Statistical analysis will be done using R version 4.0.3 (R Foundation, Vienna, Austria).

AEs will be summarized descriptively in tables and figures providing information on clinical relevance, timing, frequency, type, severity and causality by treatment arm.

A nonlinear mixed-effects (NLME) modelling will be used to determine PK parameters including absorption rate (k_a), volume of distribution (V), and clearance (CL). Concentrations are measured with a validated LC-MS/MS method^23–25. Using non linear mixed effects, the key population PK parameters will be calculated based on which an effect on the drug-drug interaction might be determined:

C_max and t_max will be observed values derived from the plasma concentration-time profile. Total drug exposure (AUC) and t_1/2 will be calculated with the NLME modeling software Monolix 2018R2 (Lixoft, Antony, France) using compartmental analysis. The elimination half-life will be estimated by the equation: t_1/2 = ln2/λ, where λ (the elimination rate constant) will be determined by performing a regression of the natural logarithm of the concentration values during the elimination period.

Ethical considerations

Independent ethics committee. The study has been reviewed and approved by the institutional research commission of the Swiss TPH, the ethics committee in Switzerland: ‘Ethikkomission Nordwest- und Zentralschweiz’ (AO_2020-00042; date of approval 24 November 2020), and the ‘Zanzibar Health Research Ethics Review Committee’ (reference no. ZAHREC/03/PR/OCT/2020/23; date of approval 22 October 2020). The study will be undertaken in accordance with the Declaration of Helsinki and good clinical practice. Material transfer agreements between the PHL-IdC and Swiss TPH will regulate the transfer of collected samples.

Evaluation of the risk-benefit ratio. Albendazole, ivermectin and moxidectin are well-known drugs and have little and mainly mild AEs as described to date (e.g., headache, abdominal pain)^{6,9,16,26–28,33}. Albendazole and ivermectin are widely used drugs in mass treatment programs against filariasis while only moxidectin is a relatively new drug, FDA-approved against onchocerciasis³⁴. All community members enrolled in the study will benefit from a clinical examination and a treatment against STHs. All participating subjects remaining positive for T. trichiura will be treated with ivermectin (200 μg/kg)/albendazole (400 mg), considering this combination showed higher efficacy compared to the existing standard treatment (albendazole alone) and the recent inclusion of ivermectin-albendazole as recommended treatment scheme against STH on the List of Essential Medicines⁴.

Subject information and consent. Information sessions at the respective schools will be conducted to explain to teachers, caregivers and potential participants the purpose and procedures of the study, as well as potential benefits and risks of participation. All parents or caregivers of eligible adolescents and all participants ≥18 years will be invited to sign a written informed consent sheet. In case the person is illiterate, an impartial witness that can read and write has to sign the consent and the illiterate participant has to give a thumb print. Parents or caregivers and adult participants will have sufficient time for reflection of their child’s or their own participation, respectively. Additionally, adolescents (aged 12–17 years) will be briefed verbally, and written assent will be sought in form of their name and signature written down or if illiterate by providing a thumbprint.

Parents or caregivers attending this meeting will receive a small provision to cover their costs for transportation (~US$ 2). Participation is voluntary and individuals have the right to withdraw from the study at any given point in time with no further obligations. Participation itself will not be awarded with compensation.

Quality control and quality assurance

We will work with a locally based external monitor, who will conduct site visits to the investigational facilities for the purpose of monitoring the study. Details will be described in a separate monitoring plan. The investigator will permit them access to study documentation and the clinical supplies dispensing and storage area. Monitoring observations and findings will be documented and communicated to appropriate study personnel and management. A corrective and preventative action plan will be requested and documented in response to any significant deviation. No sponsor-initiated audits are foreseen, but audits and inspections may be conducted by the local regulatory authorities or ethics committees. The investigator agrees to allow inspectors from regulatory agencies to review records and is encouraged to assist the inspectors in their duties, if requested.

In our study, no data and safety monitoring board will be established, since we work with well-known drugs in a small sample size and using a single dose treatment. However, advisors will be informed regularly and the findings discussed.

Dissemination of study results and publication

The final results of this study will be published in a scientific journal and presented at scientific conferences. The Bill & Melinda Gates Foundation will be acknowledged as study funder. All results from this investigation are considered confidential and shall not be made available to any third party by any member of the investigating team before publication. A summary of study conclusions will be shared with ZAHREC. After publication, study results will be made available to study communities.

Study status

The screening phase to identify eligible trial participants was initiated in March 2021.

Underlying data

No data are associated with this article.

Extended data

Open Science Framework: Efficacy and Safety of MOX/ALB vs. IVM/ALB co-administration. https://doi.org/10.17605/OSF.IO/A3N85¹⁹.

This project contains the following extended data:

Reporting guidelines

Open Science Framework: SPIRIT checklist for “Efficacy and safety of moxidectin and albendazole compared to ivermectin and albendazole co-administration in adolescents infected with Trichuris trichiura: a randomized controlled trial”. https://doi.org/10.17605/OSF.IO/A3N85¹⁹.

Data are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).