key: cord-304457-8g36h1bz authors: Idelsis, E.-M.; Jesus, P.-E.; Yaquelin, D.-R.; Dania, V.-B.; Monica, B.-R.; Lisandra, B.-R.; Jesus, C.-R.; Lisbeth, C. C.; Ernesto, P.-C.; Saily, T.-P.; Claudia, M.-S.; Ivan, C.-L.; Julio Raul, F.-M.; Hamlet, C.-R.; Marisol, D.-G.; Adriana, S.-M.; Maura, G.-S.; Sara Maria, M.-M.; Marel, A.-V.; Francisco, H.-B.; Hugo, N.-C.; Dianela, B.-G.; Abrahan, B.-C.; Mary Tania, V.-C.; Gerardo, G.-N.; Verena, M.-G.; Iraldo, B.-R. title: Effect and safety of combination of interferon alpha-2b and gamma or interferon alpha-2b for negativization of SARS-CoV-2 viral RNA. Preliminary results of a randomized controlled clinical trial. date: 2020-08-01 journal: nan DOI: 10.1101/2020.07.29.20164251 sha: doc_id: 304457 cord_uid: 8g36h1bz Abstract Objectives: IFN-alpha2b and IFN-gamma combination has demonstrated favorable pharmacodynamics for genes underlying antiviral activity which might be involved in the defense of the organism from a SARS-CoV-2 infection. Considering this we conducted a randomized controlled clinical trial for efficacy and safety evaluation of subcutaneous IFN-alpha2b and IFN-gamma administration in patients positive to SARS-CoV-2. Methods: We enrolled 19-82 years-old inpatients at the Military Central Hospital Luis Diaz Soto, Havana, Cuba. They were hospitalized after confirmed diagnosis for SARS-CoV-2 RNA by real-time reverse transcription polymerase chain reaction. Patients were randomly assigned in a 1:1 ratio to receive either, subcutaneous treatment with a co-lyophilized combination of 3.0 MIU IFN-alpha2b and 0.5 MIU IFN-gamma (HeberFERON, CIGB, Havana, Cuba), twice a week for two weeks, or thrice a week intramuscular injection of 3.0 MIU IFN-alpha2b (Heberon Alpha R, CIGB, Havana, Cuba). Additionally, all patients received lopinavir-ritonavir 200/50 mg every 12 h and chloroquine 250 mg every 12 h (standard of care). The primary endpoints were the time to negativization of viral RNA and the time to progression to severe COVID-19, from the start of treatment. The protocol was approved by the Ethics Committee on Clinical Investigation from the Hospital and the Center for the State Control of Medicines, Equipment and Medical Devices in Cuba. Informed consent was obtained from each participant. Results: A total of 79 patients with laboratory-confirmed SARS-CoV-2 infection, including symptomatic or asymptomatic conditions, fulfilled the inclusion criteria and underwent randomization. Thirty-three subjects were assigned to the HeberFERON group, and 33 to the Heberon Alpha R group. Sixty-three patients were analyzed for viral negativization, of them 78.6% in the HeberFERON group negativized the virus after 4 days of treatment versus 40.6% of patients in the Heberon Alpha R groups (p=0.004). Time to reach the negativization of the SARS-CoV-2 measured by RT-PCR in real time was of 3.0 and 5.0 days for the HeberFERON and Heberon Alpha R groups, respectively. A significant improvement in the reduction of time for negativization was attributable to HeberFERON (p=0.0027, Log-rank test) with a Hazard Ratio of 3.2 and 95% CI of 1.529 to 6.948, as compared to Heberon Alpha R treated group. Worsening of respiratory symptoms was detected in two (6.6%) and one (3.3%) patients in HeberFERON and IFN-alpha2b groups, respectively. None of the subjects transit to severe COVID-19 during the study or the epidemiological follow-up for 21 more days. RT-PCR on day 14 after the start of the treatment was negative to SARS-CoV-2 in 100% and 91% of patients of the combination of IFNs and IFN-alpha2b, respectively. Negativization for HeberFERON treated patients was related to a significant increase in lymphocytes counts and an also significant reduction in CRP as early as 7 days after commencing the therapeutic schedule. All the patients in both cohorts recover by day 14 and were in asymptomatic condition and laboratory parameters return to normal values by day 14 after treatment initiation. Adverse events were identified in 31.5% of patients, 28.5% in the control group, and 34.4% in the HeberFERON group, and the most frequent were headaches (17.4%). Conclusions: In a cohort of 63 hospitalized patients between 19 to 82 years-old with positive SARS-CoV-2, HeberFERON significantly negativized the virus on day 4 of treatment when comparing with IFN-alpha2b. Heberon Alpha R also showed efficacy for the treatment of the viral infection. Both treatments were safe and positively impact on the resolution of the symptoms. None of the patients developed severe COVID-19. Key words: COVID-19, treatment, drug, virus negativization, antiviral, interferon combination, SARS CoV-2. properties of these molecules 4 . In fact, severity of COVID-19 disease correlates with the failure to implement an IFN response to SARS-CoV-2 infection 5 . Taking these into consideration and the fact that therapeutics that target the coronavirus alone, might not be able to reverse highly pathogenic infections, the Cuban Protocol for Management of COVID-19 2 includes Heberon Alpha R and other antiviral treatment since the symptomatic phase. Cuban patients already showing symptoms or their near contacts are isolated in centers conditioned for that purpose and start receiving symptomatic treatments. As mentioned, this schedule incorporates Heberon Alpha R to lopinavir-ritonavir (Kaletra) and chloroquine (CQ). After confirmation of the positivity of SARS-CoV-2, they are hospitalized and continue to, or start to receive Heberon Alpha R, Kaletra, and CQ as established by Cuban Health Ministry guide-lines 2 . This has resulted in a favorable evolution of the patients in a cohort of 761 subjects confirmed for SARS-CoV-2 receiving Heberon Alpha R, where 95.4% fully recovered from COVID-19, with only 0.92% of case fatality rate 6 . Earlier studies of a combination of type I IFN and IFN-γ shown a synergistic inhibition of the SARSCoV virus replication in vitro 7, 8, 9, 10 . IFN-γ is a key moderator in linking the innate immunity to adaptive immune responses 11 , hence it is possible that a combinational therapy of IFNs and other antiviral drugs could significantly inhibit virus replication and modulates clinical variables related to the immune response with a positive outcome in terms of viral infection resolution 12, 13, 14 . In accordance to the previous comments we conducted this phase 2 randomized trial to establish whether a combination of IFN-α2b and gamma with the standard of care, can improve the viral load profile and clinical parameters in adults with COVID-19. Hospitalized adult patients with RT-PCR confirmed SARS-CoV-2 were enrolled in this openlabeled, single center, prospective, randomized and controlled clinical trial at Military Central Hospital "Luis Diaz Soto" Hospital, Havana, Cuba. Patients were randomly assigned to receive the combination of IFN-α2b and IFN-γ (HeberFERON, CIGB, Havana, Cuba) or IFN-α2b (Heberon Alpha R, CIGB, Havana, Cuba) based on a power of 80%, and a level of confidence set at 95%, while also considering a dropout rate of 5%. Patients were blocked randomized individually to one of two treatment arms by means of random computer-generated lists, with an allocation ratio of 1:1, with block sizes of six patients. Engineering and Biotechnology (CIGB), which has remained a product with proven antiviral efficacy and an adequate safety profile for 34 years 15 . HeberFERON (IFN-α2b and IFN-γ, colyophilized in the same vial) is produced at CIGB, and registered in Cuba for the treatment of basal cell carcinoma. 16 The study execution followed the ethical principles of the Declaration of Helsinki and the International Council for Harmonization-Good Clinical Practice guidelines. No compensation was provided for enrollment in the trial. Patient personal data were protected. (severe arterial hypertension, ischemic heart disease, diabetes mellitus, etc.), with a history of autoimmune diseases, presence of hyper inflammation syndrome, serious coagulation disorders, known hypersensitivity to any of the components of the formulation under evaluation, pregnancy or lactation, and obvious mental incapacity to issue consent and act accordingly with the study. The clinical trial protocol was approved by the Ethics Committee on Clinical Investigation of Military Central Hospital "Luis Diaz Soto", and the Center for the State Control of Medicines, Equipment and Medical Devices (CECMED) in Cuba. Patients were asked for written consent to participate after having been duly informed about the characteristics of the trial, objectives, benefits and possible risks. Likewise, they were informed of their rights to participate or not and to withdraw their consent at any time, without exposing themselves to limitations for their medical care or other retaliation. The study was registered on April 2020 at: registroclinico.sld.cu/en/trials/RPCEC00000307. After a preliminary exploratory analysis of the outcomes of the first 79 patients, the monitoring board considered a preliminary report and early publishing of the RT-PCR results from the available throat swabs in 63 patients with available throat swabs, due to the significant effect of HeberFERON on the reduction of the time to viral clearance. The trial finally included 134 patients that are now in the process of data collection for definitive processes and analysis. Data collection: Demographic, clinical, laboratory, treatments and outcome characteristics of patients were extracted from medical records and registered in to CRF and then were entered in duplicate (independently by two operators) for the subsequent process of automatic comparison and correction of the databases, necessary for statistical analysis with accurate information from the trial. However the blinding was not feasible, it was maintained for laboratory SARS-CoV-2 RNA detection by RT-PCR that is one of the endpoint of the study. The hospital received patients from several zones in Havana city diagnosed in reference centers for SARS-CoV-2 infection following the Cuban Ministry of All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . Health guidelines for diagnostic testing. Patients were defined to have SARS-CoV-2 if they had two consecutive positive results, including the confirmatory test by RT-PCR targeting amplifications of E and /or RdRP genes. A cycle threshold up to value 40 was defined as positive. Specimens were obtained from throat swabs of patients at the hospital following standard procedures and transported to a BSL2 certified laboratory at the CIGB for serial evaluation of SARS-CoV-2 viral nucleic acid detection by RT-PCR targeting after extraction by QIAamp® Viral RNA Mini kit (Qiagen, USA). A multiplexed detection by RT-PCR was carried out targeting E and/or RdPR genes plus EAV internal extraction control (TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany) as described before using Multiplex RNA Virus Master (Roche, USA). Hospital "Luis Diaz Soto" and included whole blood count, coagulation profile, serum biochemical tests (including renal and liver function, electrolytes, and coagulation), Creactive protein (induced by various inflammatory mediators such as IL-6 18 Statistical analysis: Quantitative variables were described with the arithmetic mean and its standard deviation and the median with its range. We used the absolute and relative frequency (%) for qualitative variables. The hypothesis test used was Fisher's exact test. The viral negativization analysis was performed using the Kaplan-Meier plot representation and the comparison of factors was done with the Mantel-Cox Log Rank tests. The evolution of All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . https://doi.org/10.1101/2020.07.29.20164251 doi: medRxiv preprint laboratory parameters while under treatment was analyzed using a paired mixed model (which cannot handle missing values appearing due to patient release from hospital). Correlations between virus negativization and laboratory parameters were studied using a two-tailed non-parametric Spearman correlation with 95% confidence interval. P<0.05 was considered statistically significant. Statistical analysis was performed using the Windows software package SPSS (version 25) and GraphPad Prism v8.0. We have screened 144 patients positive by RT-PCR to SARS-CoV-2. Fifty-seven patients did not fulfill the inclusion criteria, of them one with icterus, one with chronic decompensate renal insufficiency, two non-confirmed positive PCR for SARS-CoV-2, and fifty-three patients with 2 positive RT-PCR after more than 21 days of persistent virus shedding, were excluded. Patients with viral persistence were later treated with the HeberFERON out of the clinical trial (manuscript in preparation). Eight patients that did not consent were also excluded. Finally, seventy-night subjects met the inclusion criteria and were randomly assigned (1:1) to either the HeberFERON group (41 patients) or the control group (38 patients). Twelve patients did not start the treatment, 7 patients refused to start the treatment, although they have been signed the consent, and 5 were excluded due to loss of inclusion criteria. Seven patients withdrew by several causes: 3 due to worsening of respiratory symptoms (two of them in the HeberFERON group, with asthma as underlying diseases) that changed to other non-permitted in the study drug treatment; 3 patients from the control group with positive RT-PCR on day 14 were switched to receive HeberFERON out of the clinical trial by medical decision; and 1 with the appearance of an exclusion criterion (pregnancy) in the control group (see figure 1 , flow chart of the study). Four patients were not analyzed, three in the HeberFERON group, due to bad inclusions (were negative to viral RNA before the beginning of treatment as identified by the board of monitors), and one in the control group because refused the swabs sampling. Thirty and thirty-three patients were analyzed by intention to treat (ITT) in the HeberFERON and control group, respectively. Finally, swabs samples to test for viral negativization were obtained from sixty-three patients. In this cohort 29 were symptomatic (46.0%), with a median from the beginning of symptoms All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. (IQR: . Symptomatic patients with more than 7 days from the symptoms onset were more common in the HeberFERON arm (50.0%), however, these numerically differences were not statistically significant (see table 1 ). In the HeberFERON group 66.6% of symptomatic were females and in the control symptomatic males were more frequent 70.6% (p=0.024). The more common symptoms were fever and unproductive cough (16.4%), followed by headache (9.6%), decay (8.4%), odynophagia and nasal secretions (5.4%), diarrhea, dyspnea, chills and general malaise (4.1%), and others as sore throat and myalgia (2.7%). Fifty percent of patients had any comorbidity; the most frequent were hypertension (22%), asthma (6.3%), diabetes and glaucoma (4.7%). The vital signs at the time of hospital admission were not statistically different between groups. Some imbalances existed at enrollment between the groups, including a higher median age in the HeberFERON than in the control group, as well as more patients with higher than 7 days from onset of the symptoms in the HeberFERON group. No other major differences in symptoms, vital signs, laboratory results, disease severity, or treatments were observed between groups at baseline. All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . In asymptomatic patients a lower rate of negativization was observed for both IFNs. However, the HeberFERON showed a 70.6% of negativization in comparison to 46.7% for control group. The worsening of respiratory symptoms was detected in two (6.6%) and one (3.3%) patients in HeberFERON and control groups, respectively. None of the patients transit to severe COVID-19. The RT-PCR after treatment with IFNs on day 14 for hospital discharges was negative to SARS-CoV-2 in 100% and 91% of patients of HeberFERON and control cohorts, respectively. Nevertheless, the kinetics for this recovery differ between treatments groups. Earlier increase in lymphocytes percentage was observed only for HeberFERON treated patients (p=0.0141) with a marked trend for increment in lymphocytes concentrations. Also a significant decrease in CRP (p= 0.0444) was notice for this group parallel to a trend in the reduction in CPK ( Figure 3 ). The correlation between laboratory data evolution and SARS-CoV-2 virus clearance data was analyzed using a two-tailed non-parametric Spearman correlation with 95% confidence interval. Table 2 summarized the parameters identified with significant direct or indirect relation with the reduction in the time needed to achieve a negative PCR result. A particular assessment of the same parameters is also included for the symptomatic patients included for All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. Nighty-four percent of adverse events were mild and none severe. There were no differences between the incidence of any of the adverse events or duration between the treatment groups. No serious adverse events were reported. No patients died during the study (table 2) . Asymptomatic incubation period with or without detectable viral RNA, followed by nonsevere symptomatic step and viral presence, ending in a severe symptomatic stage with high viral load, characterizes the SARS-CoV-2 infection 19 , that has been widely spread. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . https://doi.org/10.1101/2020.07.29.20164251 doi: medRxiv preprint beta-1b 14 . Even 74% of negativization showed by Herberon Alpha-2b is superior to the reported by other authors 28 . Time to reach the negativization of the SARS-CoV-2 measured by RT-PCR in real-time was 3.0 and 5.0 days from the start of treatment with HeberFERON and Heberon alpha R, respectively, a difference statically significant. These results are in concordance with in vitro data about the greater sensitivity of SARS-CoV-2 to IFNs with respect to SARS-CoV 29, 30 and as compared to those treated with Heberon Alpha R. Viral dissemination is determinant in the establishment of severe disease 31 . Therefore, the shortening of time to virus clearance as has been demonstrated for HeberFERON will impact very favorable in the disease outcome in COVID-19 infected patients. The timing of initiation of antiviral therapy is another key factor in the treatment of viral infections. In the combat of SARS-CoV, no effect of several antiviral drugs was observed when the treatments were started 6-14 days after symptom onset 32 The combination of Kaletra with other antiviral agents, as has been done in SARS 37 , MERS-CoV, 38 and our trial, might enhance antiviral effects and improve clinical outcomes. The confirmation of this therapeutic approach remains to be determined. However, it has been recently shown the combination of IFNs with Kaletra is associated with more favorable clinical outcomes than the use of Kaletra alone in COVID-19 patients 35 . The presence of IFN-γ in the HeberFERON formulation additionally to its strong immune regulatory functions may restrict the angiotensin-converting enzyme 2 (ACE2) expression 39 , a receptor for cell entry for SARS-CoV-2 40 . It has been reported that this cytokine can directly inhibit viral entry for several viral infections (HCV and HIV) 41 by controlling the All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. In MERS-CoV infected mice delayed IFN treatment was associated with increased infiltration and activation of monocytes, macrophages, and neutrophils in the lungs; and enhanced pro-inflammatory cytokine expression 34 . Additionally, soon, after infection in human, application of antiviral therapy with rapid viral clearance can delay pro-inflammatory cell development, activation and their infiltration that will contribute to spar human life 14 . Delayed IFN response can also cause inflammation and tissue damage. The host may benefit from IFN presence early in the disease course, particularly when IFN system is antagonized by viral proteins or is of low competence in older aged patients 24 . An important difference between treatments concerns their effects on lymphocytes percentages among leukocytes. Only for HeberFERON treated patients a significant increase of lymphocytes percentages was observed by week 1 and this fact, as well as lymphocytes concentrations, correlates significantly with the reduction in time to virus clearance (fig 3 and table 2). All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. Herein we detected a significant reduction in CRP in patients after two administrations of HeberFERON (see figure 3) . CRP is correlated with the level of inflammation 46 , and is an important index for the diagnosis and assessment of severe pulmonary infectious diseases 50, 51 . In the early stage of COVID-19, CRP levels could reflect lung lesions and disease severity. The downregulation of CPR levels by IFNs in patients with COVID-19 early in the diseases could avoid acute inflammatory pathogenesis and disease severity 52 . Then the administration of the HeberFERON in patients primed with the antivirals may result in a further boosted antiviral effect that contribute to shortening the time for viral clearance and to lower the probabilities to develop a more severe conditions of the diseases, that implies at the end, a lower lethality rate. Although with significant more aged patients in the HeberFERON and cohort with 40% of symptomatic patients with median age of 50 years-old, none of these patients became severe ill during the trial and all of them were discharged. However, two patients from HeberFERON group worsened the respiratory symptoms. These were asymptomatic men at admission, both 33 years-old, with asthma as comorbidity that received 3 and 4 doses of HeberFERON. They were negative to SARS-CoV-2 RT-PCR since 48h and 96 h since the first HeberFERON administration. During the days of symptoms worsening climate conditions were favorable to exacerbate asthma symptomatology. They recovered in 48 hours after anti-inflammatory therapy. In the control group a symptomatic man of 80 years-old, All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . with hypertension as comorbidity, that received only one dose of Heberon Alpha R also worsened the respiratory symptoms and was transfer to ICU. His RT-PCR for viral RNA was negative 20 days after symptoms worsening and discharged. Altogether these results indicate that with high probability the rapid viral elimination detected for HeberFERON treated patients is translated into the reduction of systemic inflammation markers while inducing a significant increase in circulating lymphocytes concentrations that may explain the symptomatic improvement observed in the more risky patients in the HeberFERON group. These results adds to the anti-inflammatory effect described for IFNs in COVID-19 patients 45 , and are in correspondence with the finding of gene signature involved in Type I and Type II response in mild-to-moderate COVID-19 patients 53 . About 15% of the confirmed COVID-19 cases progress to the severe phase, with a higher risk for patients over 65 years-old 54 . Using this estimate, in the 63 patients included in our study approximately 9 patients were expected to develop severe disease; however no patients became severely ill. No death was recorded in these mild or moderate patients. In similar cohort of patients, 0.9% of mortality was described with the early use of IFNs 35 . In our trial several clinical parameters known to be related to COVID-19 progression were significantly improved by the treatments or showed a trend to a favorable behavior. These results confirm the validity of early intervention with the treatment of IFNs in patients with COVID-19, whereas demonstrated in the trial, the combination of type I and type II IFNs impacts strongly in the reduction of the risk for a severe disease likely through the efficient implementation of a timely controlled inflammatory antiviral response against the SARS-CoV-2 infection. Before being approved recently by the FDA and EMA for severe COVID-19 patients, remdesivir, was not successful in two randomized clinical trials 55, 56 . Its approval was sustained on the reduction of the illness duration in a few days 57 . Still the role of this antiviral in the inflammatory processes that drives the transit to severe and critical condition in COVID-19 patients has not been described. HeberFERON formulation that combines in one vial IFN-α2b and gamma results in an advantageous option for the treatment of COVID-19 patients. First, due to the demonstrated better pharmacodynamics 16 it is possible to administer less frequent and at lower doses than the other conventional IFNs, (IFN-α2b or IFN-β or IFN-λ) that need a thrice a week administration to have similar effect. IFN-β has been used at doses higher than 2 fold (of 12 MIU/mL 58 or 8 MIU/mL 14 ) with respect to HeberFERON doses. Second, the simultaneous All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . https://doi.org/10.1101/2020.07.29.20164251 doi: medRxiv preprint administration of both types of IFNs will promote a faster and stronger innate and adaptive immune response. At least these two facts could be responsible for the quick clearance of SARS-CoV-2 detected in our trial. It has been proposed that interferon is efficient only in patients who lacked comorbidities 59,60 ; however we have obtained a high rate of negativitazation, resolution of symptoms, and hospital discharges for HeberFERON in a cohort of patients with 57% of coexisting comorbidities. Moreover, it has been suggested that comorbidities like diabetes affect the response to IFN 60 . Two diabetic patients in our cohort negativized the virus on day 3 from the beginning of the treatment and the other at least before day 14. Our study had several limitations. This trial was open label, without a placebo group with unbalanced demographics (age years) between treatment arms. In addition, sampling methods were most likely suboptimal using the throat sampling, because of inability to do sampling of lower respiratory tract secretions. Previous studies have shown that throat-swab specimens have lower viral loads 43 . Irrespective of these limitations the HeberFERON showed efficacy and was safe in shortening virus shedding, eliminating symptoms, and discharge of patients with COVID-19. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . https://doi.org/10.1101/2020.07.29.20164251 doi: medRxiv preprint HeberFERON was a safe treatment, superior to Heberon Alpha R in shortening the time to SARS-CoV-2 viral RNA negativization in a cohort of symptomatic or asymptomatic patients between 19 and 82 years-old, with more than 95% of patients negative to the SARS-CoV-2 in 5 days of treatment. The rapid viral negativization contributes to implement an anti-inflammatory response that can protect the patients to enter in a more severe step of the disease. Early isolation combined with early administration of antiviral treatments as IFNs is an efficient approach that could contribute to save the life of patients in the COVID-19 pandemic. The use of HeberFERON might be a distinctive element in the preventive and therapeutic strategy for current or future SARS outbreaks. All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . Table 3 . Detection of SARS-CoV-2 in swabs by RT-PCR in the HeberFERON or control groups. Throat swabs were taken from 63 COVID-19 positive patients at 48h, 72h, 96h and 120h after their treatment with HeberFERON (30 patients) or standard of care (33 patients). Viral nucleic acid detection was carried out by RT-PCR resulting in positive or negative samples. For each analysis time, we represent the number of positive and negative patients, the percentage of negativization and the p value in a Fisher test analysis in an overall analysis (ALL) and splitting patients in Symptomatic (S) and Asymptomatic (A). *: p< 0.05 ; **: p> 0.01; ns: p> 0.05. All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. Median of negativization for the two treatments were also calculated. All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted August 1, 2020. . https://doi.org/10.1101/2020.07.29.20164251 doi: medRxiv preprint Leticia Martínez Hernández | internet@granma.cu.18 de junio de 2020 23:06:03. 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The New England journal of medicine Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial Remdesivir for 5 or 10 Days in Patients with Severe Covid-19 Remdesivir for the Treatment of Covid-19-Preliminary Report Efficacy and safety of interferon β-1a in treatment of severe COVID-19: A randomized clinical trial Ribavirin and interferon therapy in patients infected with the Middle East respiratory syndrome coronavirus: an observational study IFN-α2a or IFN-β1a in combination with ribavirin to treat Middle East respiratory syndrome coronavirus pneumonia: a retrospective study Baseline Characteritic and outcomes of 1591 patients infectd with SARS-CoV-2 admitted to ICU of the Lombardy Regii The authors were responsible for designing the trial and for collecting and analyzing the data.The authors assured the completeness and accuracy of the data collection and the adherence to the protocol. The details about the trial are provided in the protocol that has been posted in TRIALS 17 and is in processing by the editors of the journal.The primary endpoints were the time to viral RNA negativization from the start of treatment and the time to progression to severe COVID-19.