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RNA Virus InfectionsACTEarlyNCT07254052

Effectiveness of Ammonium Chloride in Reducing Viral Load

An observational study of RNA Virus Infections and Viral Infection COVID-19, run by National and Kapodistrian University of Athens at 2 sites in Greece.

Completed

Finished. The study has ended and is not enrolling. Results may be posted on the official record.

Official record
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Trial summary

This is a prospective, double-blind, randomized, comparative effectiveness pilot study evaluating the effect of a sustained-release ammonium chloride formulation (ACF) on viral load dynamics in adult patients with mild or moderate COVID-19 or seasonal influenza infection. Read more about this study

Who is it for?
Ages 18 years and older
Type of treatment
Pill or oral medicine
How many people
32 participants
Where
2 sitesGreece
Study dates
1 Sep 2024to 30 May 2025
Phase
Observational
Are there results?
None posted in the registry

Key requirements

Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.

  • Ages 18 years and older
  • All sexes
  • No healthy volunteers
  • Adult, Older adult

Study population: Adult patients (≥18 years old) with mild or moderate COVID-19 or seasonal influenza infection were recruited at the Outpatients Clinic and the Emergency Department of Sotiria General Hospital for Chest Diseases in Athens, Greece and at "En Ygeia" outpatient clinic. Both inpatients and outpatients were eligible. All participants had a laboratory-confirmed diagnosis by rapid antigen testing and were enrolled within 3-5 days of symptom onset. Patients with severe disease, organ transplantation, pregnancy, or frailty score ≥5 were excluded. A total of 32 participants completed the study procedures.

Who may be able to join

  • Adults ≥ 18 years.
  • Positive rapid test for Sars-CoV-2 19 or influenza (performed at the Outpatients Clinic or the Emergency Department).
  • Informed consent.

Who cannot join

  • Age < 18 years.
  • Pregnancy, and/or lactation.
  • Denial of Informed Consent.
  • Known allergy to ammonium chloride or to any of the ACF, or VDF excipients.
  • Organ transplantation.
  • Frailty score ≥ 5.

Trial locations and contacts

2 sites in 1 country.

Greece2 sites

  • En Ygeia Clinic

    Athens, 11524

  • Sotiria General Hospital

    Athens

Other trials in Greece

About this study

Official title: Effectiveness of a Sustained-release Ammonium Chloride Formulation in Reducing the Viral Load of Patients With COVID-19 or Influenza

This is a prospective, double-blind, randomized, comparative effectiveness pilot study evaluating the effect of a sustained-release ammonium chloride formulation (ACF) on viral load dynamics in adult patients with mild or moderate COVID-19 or seasonal influenza infection.

Eligible participants are randomly assigned in a 1:1 ratio to receive either ACF (ammonium chloride with vitamin D) or a control formulation (vitamin D only) twice daily for 10 consecutive days. The primary objective is to assess the reduction of viral load measured by RT-PCR cycle threshold (Ct) values at baseline, Day 3-5, and Day 10-12. Secondary objectives include the duration and severity of symptoms, incidence of hospitalization, oxygen supplementation, ICU admission, and mortality.

The study is conducted in both inpatient and outpatient settings at Sotiria General Hospital for Chest Diseases, Athens, Greece as well as at the outpatient clinic "En Ygeia". All viral load analyses are performed at the Department of Pharmacy, National and Kapodistrian University of Athens. A total of 32 participants were enrolled. This study aims to generate early clinical evidence regarding the antiviral effectiveness of ACF as an adjunct strategy in respiratory RNA virus infections.

Read the detailed description

This is a prospective, double-blind, randomized, comparative effectiveness pilot clinical study designed to evaluate the antiviral effect of a sustained-release ammonium chloride formulation (ACF) in adult patients with mild or moderate SARS-CoV-2 or seasonal influenza infection. Respiratory viral infections remain a major global health burden, and safe, accessible adjunct therapies are needed to complement existing public health measures and vaccination strategies.

Eligible participants are adults (≥18 years) with a positive rapid test for SARS-CoV-2 or influenza A/B, recruited at the Outpatients Clinic or Emergency Department of Sotiria General Hospital for Chest Diseases and at "En Ygeia" outpatient clinic. Participants are randomized in a 1:1 ratio to receive either:

  1. ACF (nutritional supplement): sustained-release ammonium chloride formulation with vitamin D, one capsule every 12 hours for 10 days
  2. VDF (control): matched sustained-release placebo formulation containing only vitamin D, one capsule every 12 hours for 10 days.

Both inpatients and outpatients are included. Exclusion criteria are: age <18 years, pregnancy or lactation, denial of informed consent, known allergy to study components, organ transplantation, and frailty score ≥5. Participants may discontinue the study in cases of intolerance, non-compliance, or withdrawal of consent.

The primary endpoint is the reduction of viral load, assessed through RT-PCR cycle threshold (Ct) values obtained at baseline (Day 1), Day 3-5, and Day 10-12.

Secondary endpoints include:

  • Duration and severity of symptoms for 30 days
  • Incidence of hospitalization (for outpatients)
  • Need for oxygen supplementation
  • ICU admission
  • Intubation
  • Death. All RT-PCR testing for SARS-CoV-2 and influenza A/B were performed at the Department of Pharmacy, National and Kapodistrian University of Athens. The study employs standardized sampling, nucleic acid extraction, and real-time PCR quantification with certified reference materials.

Statistical analysis compared Ct values across timepoints within and between groups. Mean percentage change in Ct values was compared using appropriate parametric or non-parametric tests, with additional analyses for secondary outcomes. The final sample size was 32 patients.

This pilot trial aims to provide early clinical evidence regarding the potential antiviral effectiveness of ammonium chloride in the context of mild respiratory viral infections, contributing to the design of larger confirmatory studies.

Keywords
ammonium chloride, influenza, SARS-CoV-2, RNA virus infections, virus clearance
Medical subject headings
RNA Virus Infections, Influenza, Human

What participants receive

What participants receive. In a randomised study each person is assigned to one group by chance.

Study groups in NCT07254052
Study groupRoleWhat they receive
ACF receiving participantsParticipants assigned to this group received the sustained-release ammonium chloride formulation (ACF), containing 500 mg ammonium chloride and 2,000 IU vitamin D per capsule. One capsule was administered orally every 12 hours for 10 consecutive days, starting on Day 1. Both inpatients and outpatients with mild or moderate COVID-19 or seasonal influenza infection were included. The intervention aimed to evaluate the effect of ammonium chloride on viral load reduction and clinical outcomes compared to the control group. Randomization and double-blinding were maintained throughout the study.
Control Group (VDF receiving participants)Participants assigned to this group received the control formulation consisting of sustained-release tablets containing 2,000 IU vitamin D and identical excipients, but no ammonium chloride. One capsule was administered orally every 12 hours for 10 consecutive days, starting on Day 1. Both inpatients and outpatients with mild or moderate COVID-19 or seasonal influenza infection were included. This control arm was used to compare changes in viral load dynamics and clinical outcomes against the intervention group. Randomization and double-blinding were maintained throughout the study.

What is being measured

The primary outcome is the main question the study is designed to answer.

  1. Change in viral load measured by RT-PCR Ct values

    Change in viral load was assessed through real-time PCR (RT-PCR) cycle threshold (Ct) values for SARS-CoV-2 or influenza A/B from nasopharyngeal swabs. Measurements were performed at three predefined time points (Day 1, Day 3-5, and Day 10-12). Viral load reduction was defined as an increase in Ct values over time. Samples were processed at the Department of Pharmacy, National and Kapodistrian University of Athens.

    Measured: Baseline (Day 1), Day 3-5, Day 10-12

Secondary outcomes (6)

  1. Duration of symptoms

    Duration of respiratory and systemic symptoms was recorded for 30 days after diagnosis. Data were collected through patient interviews and medical record review.

    Measured: Up to Day 30 after diagnosis

  2. Hospitalization rate (outpatients only)

    Number of outpatient participants requiring hospitalization within 30 days of diagnosis.

    Measured: Up to Day 30

  3. Need for oxygen administration

    Number of participants requiring oxygen therapy during follow-up.

    Measured: Up to Day 30

  4. ICU admission

    Number of participants admitted to the ICU for respiratory deterioration.

    Measured: Up to Day 30

  5. Intubation

    Number of participants requiring endotracheal intubation.

    Measured: Up to Day 30

  6. Mortality

    Number of deaths from any cause during the 30-day follow-up period.

    Measured: Up to Day 30

Timeline

From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.

  1. First posted28 Nov 2025
  2. Study start1 Sep 2024
  3. Primary completion30 Apr 2025
  4. Study completion30 May 2025

Resources

More details

For doctors and researchers: how the study is designed, who runs it, and its registry IDs.

Study design
Study type
Observational
Observational model
Case-control
Time perspective
Prospective
Enrolment
32 participants (actual)
IDs, dates and publications
ClinicalTrials.gov ID
NCT07254052
Sponsor's protocol ID
Sotiria GH 14328/17-05-2025
Acronym
ACTEarly
Status last verified
Nov 2025
First submitted
19 Nov 2025
Last updated
28 Nov 2025
Results posted
No
Data monitoring committee
No
FDA-regulated drug
No
FDA-regulated device
No
Individual data sharing
No

Publications

  • Zeichhardt H, Wetz K, Willingmann P, Habermehl KO. Entry of poliovirus type 1 and Mouse Elberfeld (ME) virus into HEp-2 cells: receptor-mediated endocytosis and endosomal or lysosomal uncoating. J Gen Virol. 1985 Mar;66 ( Pt 3):483-92. doi: 10.1099/0022-1317-66-3-483.

    PubMed 2857762

  • Yoshimura A, Ohnishi S. Uncoating of influenza virus in endosomes. J Virol. 1984 Aug;51(2):497-504. doi: 10.1128/JVI.51.2.497-504.1984.

    PubMed 6431119

  • Superti F, Seganti L, Orsi N, Divizia M, Gabrieli R, Pana A. The effect of lipophilic amines on the growth of hepatitis A virus in Frp/3 cells. Arch Virol. 1987;96(3-4):289-96. doi: 10.1007/BF01320970.

    PubMed 2821967

  • Silva PJS, Sagastizabal C, Nonato LG, Struchiner CJ, Pereira T. Optimized delay of the second COVID-19 vaccine dose reduces ICU admissions. Proc Natl Acad Sci U S A. 2021 Aug 31;118(35):e2104640118. doi: 10.1073/pnas.2104640118.

    PubMed 34408076

  • Shrestha LB, Foster C, Rawlinson W, Tedla N, Bull RA. Evolution of the SARS-CoV-2 omicron variants BA.1 to BA.5: Implications for immune escape and transmission. Rev Med Virol. 2022 Sep;32(5):e2381. doi: 10.1002/rmv.2381. Epub 2022 Jul 20.

    PubMed 35856385

  • Shi T, Denouel A, Tietjen AK, Campbell I, Moran E, Li X, Campbell H, Demont C, Nyawanda BO, Chu HY, Stoszek SK, Krishnan A, Openshaw P, Falsey AR, Nair H; RESCEU Investigators. Global Disease Burden Estimates of Respiratory Syncytial Virus-Associated Acute Respiratory Infection in Older Adults in 2015: A Systematic Review and Meta-Analysis. J Infect Dis. 2020 Oct 7;222(Suppl 7):S577-S583. doi: 10.1093/infdis/jiz059.

    PubMed 30880339

  • Shang C, Zhuang X, Zhang H, Li Y, Zhu Y, Lu J, Ge C, Cong J, Li T, Tian M, Jin N, Li X. Inhibitors of endosomal acidification suppress SARS-CoV-2 replication and relieve viral pneumonia in hACE2 transgenic mice. Virol J. 2021 Feb 27;18(1):46. doi: 10.1186/s12985-021-01515-1.

    PubMed 33639976

  • Prabhakara C, Godbole R, Sil P, Jahnavi S, Gulzar SE, van Zanten TS, Sheth D, Subhash N, Chandra A, Shivaraj A, Panikulam P, U I, Nuthakki VK, Puthiyapurayil TP, Ahmed R, Najar AH, Lingamallu SM, Das S, Mahajan B, Vemula P, Bharate SB, Singh PP, Vishwakarma R, Guha A, Sundaramurthy V, Mayor S. Strategies to target SARS-CoV-2 entry and infection using dual mechanisms of inhibition by acidification inhibitors. PLoS Pathog. 2021 Jul 12;17(7):e1009706. doi: 10.1371/journal.ppat.1009706. eCollection 2021 Jul.

    PubMed 34252168

  • Pifarre I Arolas H, Acosta E, Lopez-Casasnovas G, Lo A, Nicodemo C, Riffe T, Myrskyla M. Years of life lost to COVID-19 in 81 countries. Sci Rep. 2021 Feb 18;11(1):3504. doi: 10.1038/s41598-021-83040-3.

    PubMed 33603008

  • Nguyen NN, Houhamdi L, Hoang VT, Stoupan D, Fournier PE, Raoult D, Colson P, Gautret P. High rate of reinfection with the SARS-CoV-2 Omicron variant. J Infect. 2022 Aug;85(2):174-211. doi: 10.1016/j.jinf.2022.04.034. Epub 2022 Apr 23. No abstract available.

    PubMed 35472367

  • Mizzen L, Hilton A, Cheley S, Anderson R. Attenuation of murine coronavirus infection by ammonium chloride. Virology. 1985 Apr 30;142(2):378-88. doi: 10.1016/0042-6822(85)90345-9.

    PubMed 2997991

  • Maltezou HC, Raftopoulos V, Vorou R, Papadima K, Mellou K, Spanakis N, Kossyvakis A, Gioula G, Exindari M, Froukala E, Martinez-Gonzalez B, Panayiotakopoulos G, Papa A, Mentis A, Tsakris A. Association Between Upper Respiratory Tract Viral Load, Comorbidities, Disease Severity, and Outcome of Patients With SARS-CoV-2 Infection. J Infect Dis. 2021 Apr 8;223(7):1132-1138. doi: 10.1093/infdis/jiaa804.

    PubMed 33388780

  • Maltezou HC, Papanikolopoulou A, Vassiliu S, Theodoridou K, Nikolopoulou G, Sipsas NV. COVID-19 and Respiratory Virus Co-Infections: A Systematic Review of the Literature. Viruses. 2023 Mar 28;15(4):865. doi: 10.3390/v15040865.

    PubMed 37112844

  • Maltezou HC, Papadima K, Gkolfinopoulou K, Ferentinos G, Mouratidou E, Andreopoulou A, Pavli A, Magaziotou I, Georgakopoulou T, Mellou K, Vorou R, Antoniadou A, Stathakarou A, Chrysochoou A, Gogos C, Karaiskou A, Kotanidou A, Koutsoukou A, Marangos M, Mentis A, Metallidis S, Papa A, Pefanis A, Tsakris A, Sipsas NV. Coronavirus disease 2019 pandemic in Greece, February 26 - May 3, 2020: The first wave. Travel Med Infect Dis. 2021 May-Jun;41:102051. doi: 10.1016/j.tmaid.2021.102051. Epub 2021 Apr 2. No abstract available.

    PubMed 33819570

  • Maltezou HC, Krumbholz B, Mavrouli M, Tseroni M, Gamaletsou MN, Botsa E, Anastassopoulou C, Gikas A, Fournarakou E, Kavieri M, Koureli A, Mandilara D, Marinopoulou A, Theodorikakou A, Tsiahris P, Zarzali A, Pournaras S, Lourida A, Elefsiniotis I, Vrioni G, Sipsas NV, Tsakris A. A study of the evolution of the third COVID-19 pandemic wave in the Athens metropolitan area, Greece, through two cross-sectional seroepidemiological surveys: March, June 2021. J Med Virol. 2022 Apr;94(4):1465-1472. doi: 10.1002/jmv.27465. Epub 2021 Dec 4.

    PubMed 34812522

Questions about this trial

What is this clinical trial studying?

An observational study of RNA Virus Infections and Viral Infection COVID-19, run by National and Kapodistrian University of Athens at 2 sites in Greece.

The full registered description is in the About this study section and the official record.

Who may be eligible to take part?

The sponsor lists 3 inclusion and 6 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.

Is this clinical trial currently recruiting?

The record says: finished, as of the sponsor's last update on 28 Nov 2025. Status can change between updates, so confirm with the study contact before making plans.

Where is the study taking place?

The record lists 2 sites in Greece. Whether a site is taking part right now should be confirmed with that site. See every site.

What are the possible benefits and risks?

They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.

Are results available for this study?

The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.

How can a doctor refer a patient or contact the research team?

The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.

What does the primary outcome measure?

Change in viral load measured by RT-PCR Ct values (time frame: Baseline (Day 1), Day 3-5, Day 10-12). It is what the study was designed to measure, not a finding.

Where can I read the original study information?

On the ClinicalTrials.gov record for NCT07254052, which also lists 28 linked publications.

Does a listing mean the treatment is approved or proven to work?

No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.

How often is this information updated?

AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 28 Nov 2025.

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