RESEARCH ON CUBAN INTERFERON ALFA-2b

Cuban interferon alpha-2b. Thirty years as an effective and safe drug. Nodarse Cuní H, López Saura PA. Biotecnol Apl. 2017;34(1):1211–7. (aquí para versión en español)
Since 1986, the Center for Genetic Engineering and Biotechnology of Havana, Cuba, has produced the recombinant human interferon alpha-2b, marketed as Heberon® Alfa R. The therapeutic use of this product has accumulated a high number of researches carried out in the country’s public health network. For the analysis of its safety profile, 28 years of reports of adverse events were reviewed in 5806 individuals, both children and adults, coming from 147 clinical trials or healthy assistances using the product. This review also contains a safety comparison between lyophilized and liquid formulations. In addition, an analysis of the connection between the occurrence of adverse events and the demographic characteristics of the patients, an analysis of immunogenicity and another on the variation of the thyroid function associated to the use of Heberon® Alfa R were included. Finally, a general analysis of the product’s efficacy based on the number of treated patients and the clinical results obtained are presented. Adverse events were reported in 4864 subjects (84 %). The main adverse events were those corresponding to the flu-like syndrome, with higher frequency in male white patients. Hypothyroidism and immunogenicity behaved lower than similar products in the international pharmaceutical market. Approximately 60 % of the treated patients obtained a relevant therapeutic response and a liquid formulation offered a better benefit/ risk ratio. The extensive clinical information evaluated recognizes a Heberon® Alfa R as a safe and effective drug, 30 years after its first production.

Clinical immunogenic profile of Heberon® alpha. Nodarse-Cuní H, Bermúdez-Badel CH, García García I, Bello Rivero I, Pedro López-Saura. Rev Cubana Farm. 2017 Jul;51(3). Spanish.
The analysis of the immunogenicity profile of recombinant human interferon alfa-2b was performed in 952 patients from 23 clinical studies, performed with the lyophilized formulation with albumin. The experimental design was started with a “sandwich” type enzyme-linked immunosorbent assay (ELISA) for detecting the presence of the antibody and its confirmation of specificity (IFN alpha-2b/sample/conjugate protein A-peroxidase), and then we investigated, through a biological assay in cells, the ability of the antibody to neutralize the antiviral activity of interferon alpha.  The development of antibodies with neutralizing capacity for the antiviral action occurred in 22 patients, representing 2.3% of those evaluated. This percentage of immunogenicity of recombinant human interferon alfa-2b, produced by the Center for Genetic Engineering and Biotechnology, is below the reported 2.7% for recombinant human interferon alfa-2b and 25.7% for interferon alfa-2a in the international market.  Recombinant human interferon alfa-2b produced at the Center for Genetic Engineering and Biotechnology can be used as a safe drug for the treatment of all diseases included in its therapeutic indications.

Interferon alpha-2b and ribavirin as combined therapy for chronic hepatitis C in Cuba: National Program.  Nodarse-Cuní H, Arus-oler E, Rivera-Reimón LL, Pérez-Lorenzo M, Samada-Suárez M, García-Ferrera WO, et al. Biotecnol Apl. 2012;29(3):184–8. Spanish.
The treatment of chronic hepatitis C with interferon alpha (IFN-α) is widely used. However, the relapse rate is high, and sustained response is only in 10-20%. A combined treatment based on the synergic antiviral effects described for IFN-α and ribavirin was used as a National Program in Cuba. The study enrolled 357 patients treated during 48 weeks with an injection of IFN-α, 3 times weekly, combined with oral ribavirin in daily doses, the doses adjusted to body weight. Sustained virological response was the efficacy end point, supported by biochemical and histological changes. Normalization in transaminase levels occurs in 60.5% of patients after the first 4 weeks, 71.4% at 26 weeks and 60.2% at the end of treatment. In similar moments, the viral load was undetectable in 42.9%, 42.6% and 37.0% respectively. The implementation of this National Program led to 49.0% and 29.7% of biochemical and virological sustained response respectively. A histological improvement was observed in 53.5% of evaluated patients. The treatment was well tolerated and almost all adverse reactions were attributable to IFN-α. The main adverse reports were: anemia, leucopenia, asthenia, fever, headache, arthralgias, anorexia and myalgia. Anti-interferon antibodies were developed in 38 patients, in 3 of them as neutralizing of antiviral activity. These results confirm the efficacy and security profile of both drugs as combined therapy for the chronic hepatitis C and represent the first clinical data generated from its extensive use in the Cuban general population. The virological response was in agreement with international reports for populations with similar characteristics.

Interferon alpha-2b as adjuvant treatment of recurrent respiratory papillomatosis in Cuba: National Programme (1994–1999 report). Nodarse-Cuní H, Iznaga-Marín N, Viera-Álvarez D, Rodríguez-Gómez H, Fernández-Fernández H, Blanco-López Y, et al. J Laryngol Otol. 2004 Sep;118(9):681–7.
Respiratory papillomatosis is a life-spoiling disease due to its high recurrence rate. Interferon (IFN) alpha-2b treatment, adjuvant to surgery, was assessed for its contribution to disease control and patient quality of life improvement. One hundred and sixty-nine patients (85 children and 84 adults) were included after surgical removal of the lesions followed by intramuscular IFN alpha-2b (Heberon alfa R, Heber Biotec), starting with 105 IU/Kg weight in children or 6 × 106 IU in adults, three times per week. The dose was reduced monthly, if no relapses occurred, until a monthly maintenance with 5 × 104 IU/Kg of weight in children or 3 × 106 IU in adults up to two years. In case of relapse, it was surgically removed and the patient returned to the higher dose level. The relapse frequency decreased significantly in 77 per cent (69/90) of the recurrent patients both in children (34/46, 74 per cent) and adults (35/44, 79 per cent). Among patients included after their first papilloma, 67 per cent (44/66) had complete (no relapses) or partial (only one relapse) responses (children: 15/33, 45 per cent; adults 29/33, 88 per cent). One hundred and eighteen patients (73 per cent) concluded the treatment without lesions (children: 58 per cent; adults 82 per cent), while the rest showed a significant reduction in the number and size of lesions. IFN was well tolerated. Sixty-two patients (38 per cent) did not have adverse events. The main adverse reactions were fever (59 per cent), chills (24 per cent), arthralgias and myalgias (14 per cent) and headache (10 per cent). One patient developed anti-IFN alpha neutralizing antibodies and became resistant to treatment with recombinant IFN alpha-2b; he responded to natural leucocyte IFN alpha. Treatment with IFN alpha-2b, as an adjuvant to surgery represents a favourable and safe therapeutic alternative for patients with recurrent respiratory papillomatosis.

Molecular Characterization of Recombinant Human Interferon Alpha-2b Produced in Cuba. Santana H, Martínez E, Sánchez JC, Moya G, Sosa R, Hardy E, et al. Biotecnol Apl. 1999;16(3)154–9.
The recombinant human interferon alpha 2b (IFN-a 2b) produced by Cuban technology is obtained from Escherichia coli. This is the active principle of the product registered in Cuba as Heberon alfa R® trademark (IFN-alpha 2b, CIGB, Havana), which has been successfully used worldwide for the therapy of several viral diseases and neoplasms. Here we describe the purity and identity tests used for its molecular characterization. The data show a product with a well-established identity, a high purity and a specific activity higher than 1.4 x 108 IU/mg of proteins. We also compared the final preparation with other IFN-alpha 2 products available in the international market. It behaved very similar to Intron A® and Roferon A®, and showed a higher homogeneity when compared with Bioferon® and Interimmun®.

RESEARCH ON INTERFERONS FOR CORONAVIRUS INFECTIONS

COVID19 Treatment: Close to a Cure? A Rapid Review of Pharmacotherapies for the Novel Coronavirus. Song Y, Zhang M, Yin L, Wang K, Zhou Y, Zhou M, et al. Preprints. 2020 Mar 26. DOI: 10.20944/preprints202003.0378.v1.
Currently, there is no specific treatment for COVID-19 proven by clinical trials. WHO and CDC guidelines therefore endorse supportive care only. However, frontline clinicians have been applying several virus-based and host-based therapeutics in order to combat SARS-CoV-2. Medications from COVID-19 case reports, observational studies and the COVID-19 Treatment Guideline issued by the China’s National Health Commission (7^th edition published March 3^rd, 2020. Edited translation attached) are evaluated in this review. Key evidence from relevant in vitro researches, animal models and clinical studies in SARS-CoV-2, SARS-CoV and MERS-CoV are examined. Antiviral therapies remdesivir, lopinavir/ritonavir and umifenovir, if considered, could be initiated before the peak of viral replication for optimal outcomes. Ribavirin may be beneficial as an add-on therapy and is ineffective as a monotherapy. Corticosteroids use should be limited without indicating comorbidities. IVIG is not recommended due to lack of data in COVID-19. Xuebijing may benefit patients with complications of bacterial pneumonia or sepsis. The efficacy of interferon is unclear due to conflicting outcomes in SARS and MERS studies. Chloroquine and hydroxychloroquine have shown in vitro inhibition of SARS-CoV-2 and may be beneficial as both prophylactic and treatment therapy. For patients who developed cytokine release syndrome, interleukin-6 inhibitors may be beneficial. Given the rapid disease spread and increasing mortality, active treatment with readily available medications may be considered timely prior to disease progression.

Discovering drugs to treat coronavirus disease 2019 (COVID-19). Dong L, Hu S, Gao J. Drug Discov Therap. 2020 Feb 20;14(1):58–60.
The SARS-CoV-2 virus emerged in December 2019 and then spread rapidly worldwide, particularly to China, Japan, and South Korea. Scientists are endeavoring to find antivirals specific to the virus. Several drugs such as chloroquine, arbidol, remdesivir, and favipiravir are currently undergoing clinical studies to test their efficacy and safety in the treatment of coronavirus disease 2019 (COVID-19) in China; some promising results have been achieved thus far. This article summarizes agents with potential efficacy against SARS-CoV-2.

Antiviral drugs specific for coronaviruses in preclinical development. Odedeji AO, Sarafianos SG. Curr Opinion Virol. 2014 Oct;8:45–53.
Coronaviruses are positive stranded RNA viruses that cause respiratory, enteric and central nervous system diseases in many species, including humans. Until recently, the relatively low burden of disease in humans caused by few of these viruses impeded the development of coronavirus specific therapeutics. However, the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV), and more recently, Middle East respiratory syndrome coronavirus (MERS-CoV), has impelled the development of such drugs. This review focuses on some newly identified SARS-CoV inhibitors, with known mechanisms of action and their potential to inhibit the novel MERS-CoV. The clinical development of optimized versions of such compounds could be beneficial for the treatment and control of SARS-CoV, the current MERS-CoV and other future SARS-like epidemics.

Development of the cDNA chip for SARS virus and a primary study on the possible molecular mechanism of interferon alpha 2b inhibiting the SARS virus replication. Shu YL, Duan ZJ, Wang Z, Sun MS, Zhang J, Zhang LL, et al. Chinese J Exp Clin Virol. 2003 Aug;17(3):209–12.
To study the molecular mechanism of interferon alpha 2b (IFN alpha 2b) inhibiting the SARS virus replication. SARS-associated coronavirus (SARS virus) cDNA chip was developed and applied to detect the virus RNA transcription levels in the interferon-treated and untreated cell cultures, and the mechanism of anti-SARS virus activity of interferon alpha 2b in cell culture system was explored. SARS virus cDNA chip was prepared by comparing the published SARS virus genome sequence, and the cDNA chip was used to study the interferon alpha2b function during SARS virus replication. SARS virus cDNA chip was successfully prepared by using PCR method. The results showed that the cDNA chip could be used to detect the viral RNA transcription level. Interferon alpha 2b could inhibit almost all the SARS virus gene transcription. An unknown gene at the position 28130-28426 bp, named as U gene, may play an important role during the viral replication. A SARS virus whole genome cDNA chip was established. It could be used to study the virus molecular biology and antiviral drug screening. The results also showed that interferon alpha 2b could inhibit almost the whole virus gene transcription by using the cDNA chip.

Inhibition of novel β coronavirus replication by a combination of interferon-α2b and ribavirin. Falzarano D, de Wit E, Martellaro C, Callison J, Munster VJ, FeldmannH. Sci Rep. 2013 Apr 18;3:1686. DOI: 10.1038/srep01686.
The identification of a novel β coronavirus, nCoV, as the causative agent of severe respiratory illness in humans originating in Saudi Arabia, Qatar and Jordan has raised concerns about the possibility of a coronavirus pandemic similar to that of SARS-CoV. As a definitive treatment regimen has never been thoroughly evaluated for coronavirus infections, there is an urgent need to rapidly identify potential therapeutics to address future cases of nCoV. To determine an intervention strategy, the effect of interferon-α2b and ribavirin on nCoV isolate hCoV-EMC/2012 replication in Vero and LLC-MK2 cells was evaluated. hCoV-EMC/2012 was sensitive to both interferon-α2b and ribavirin alone in Vero and LLC-MK2 cells, but only at relatively high concentrations; however, when combined, lower concentrations of interferon-α2b and ribavirin achieved comparable endpoints. Thus, a combination of interferon-α2b and ribavirin, which are already commonly used in the clinic, may be useful for patient management in the event of future nCoV infections.

Inhibition of SARS Coronavirus Infection in Vitro with Clinically Approved Antiviral Drugs. Tan E, Ooi EE, Lin CY, Tan HC. Emerg Infect Dis. 2004 May;10(4):581–6.
Severe acute respiratory syndrome (SARS) is an infectious disease caused by a newly identified human coronavirus (SARS-CoV). Currently, no effective drug exists to treat SARS-CoV infection. In this study, we investigated whether a panel of commercially available antiviral drugs exhibit in vitro anti–SARS-CoV activity. A drug-screening assay that scores for virus-induced cytopathic effects on cultured cells was used. Tested were 19 clinically approved compounds from several major antiviral pharmacologic classes: nucleoside analogs, interferons, protease inhibitors, reverse transcriptase inhibitors, and neuraminidase inhibitors. Complete inhibition of cytopathic effects of SARS-CoV in culture was observed for interferon subtypes, β-1b, α-n1, α-n3, and human leukocyte interferon α. These findings support clinical testing of approved interferons for the treatment of SARS.

Interferon- β 1a and SARS Coronavirus Replication. Hensley LE, Fritz EA, Jahrling PB, Karp CL, Huggins JW, et al. Emerg Infect Dis. 2004 Feb;10(2):317–9.
A global outbreak of severe acute respiratory syndrome (SARS) caused by a novel coronavirus began in March 2003. The rapid emergence of SARS and the substantial illness and death it caused have made it a critical public health issue. Because no effective treatments are available, an intensive effort is under way to identify and test promising antiviral drugs. Here, we report that recombinant human interferon (IFN)-β 1a potently inhibits SARS coronavirus replication in vitro.

Repurposing of Clinically Developed Drugs for Treatment of Middle East Respiratory Syndrome Coronavirus Infection. Dyall J, Coleman CM, Hart BJ, Venkataraman T, Holbrook MR, Kindrachuk J, et al. Antimicrob Agents Chemother. 2014 Aug;58(8)4885–93.
Outbreaks of emerging infections present health professionals with the unique challenge of trying to select appropriate pharmacologic treatments in the clinic with little time available for drug testing and development. Typically, clinicians are left with general supportive care and often untested convalescent-phase plasma as available treatment options. Repurposing of approved pharmaceutical drugs for new indications presents an attractive alternative to clinicians, researchers, public health agencies, drug developers, and funding agencies. Given the development times and manufacturing requirements for new products, repurposing of existing drugs is likely the only solution for outbreaks due to emerging viruses. In the studies described here, a library of 290 compounds was screened for antiviral activity against Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus (SARS-CoV). Selection of compounds for inclusion in the library was dependent on current or previous FDA approval or advanced clinical development. Some drugs that had a well-defined cellular pathway as target were included. In total, 27 compounds with activity against both MERS-CoV and SARS-CoV were identified. The compounds belong to 13 different classes of pharmaceuticals, including inhibitors of estrogen receptors used for cancer treatment and inhibitors of dopamine receptor used as antipsychotics. The drugs identified in these screens provide new targets for in vivo studies as well as incorporation into ongoing clinical studies.

Ribavirin and interferon-α2b as primary and preventive treatment for Middle East respiratory syndrome coronavirus: a preliminary report of two cases. Khalid M, Al Rabiah F, Mobeireek A, Butt TS, Mutairy EA. Antiviral Ther. 2014 May;20(1). DOI: 10.3851/IMP2792.
Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized transmissible viral infection with high virulence and case fatality rates for which there is no currently defined primary treatment or prophylaxis. Saudi Arabia has the largest reported number of cases so far. Like severe acute respiratory syndrome (SARS), MERS is caused by a coronavirus. Combination therapy with interferon-α2b and ribavirin has been used successfully as primary treatment and prophylaxis in SARS. Because of similarities between the two coronaviruses, treatment with ribavarin and interferon-α2b has been suggested as a potential therapy for MERS-CoV. Studies in animal models of MERS-CoV have shown the combination of ribavirin and interferon-α2b to be effective both as primary treatment and prophylaxis. In this report, we describe for the first time use of this combination as a primary treatment for a patient with MERS-CoV infection and as prophylaxis for his spouse and discuss its possible role.

Treatment with interferon α-2b and ribavirin improves outcomes in MERS-CoV-infected rhesus macaques. Falzarano D, de Wit E, Rasmussen AL, Feldmann F, Okumura A, Scott DP, et al. Nat Med. 2013 Oct;19(10):1313–7.  The emergence of Middle East respiratory syndrome coronavirus (MERS-CoV) is of global concern: the virus has caused severe respiratory illness, with 111 confirmed cases and 52 deaths at the time of this article’s publication. Therapeutic interventions have not been evaluated in vivo; thus, patient management relies exclusively on supportive care, which, given the high case-fatality rate, is not highly effective. The rhesus macaque is the only known model organism for MERS-CoV infection, developing an acute localized to widespread pneumonia with transient clinical disease that recapitulates mild to moderate human MERS-CoV cases. The combination of interferon-α2b and ribavirin was effective in reducing MERS-CoV replication in vitro; therefore, we initiated this treatment 8 h after inoculation of rhesus macaques. In contrast to untreated, infected macaques, treated animals did not develop breathing abnormalities and showed no or very mild radiographic evidence of pneumonia. Moreover, treated animals showed lower levels of systemic (serum) and local (lung) proinflammatory markers, in addition to fewer viral genome copies, distinct gene expression and less severe histopathological changes in the lungs. Taken together, these data suggest that treatment of MERS-CoV infected rhesus macaques with IFN-α2b and ribavirin reduces virus replication, moderates the host response and improves clinical outcome. As these two drugs are already used in combination in the clinic for other infections, IFN-α2b and ribavirin should be considered for the management of MERS-CoV cases.

Unravelling the convoluted biological roles of type I interferons (IFN-Is) in infection and immunity: a way forwad for the therapeutics and vaccine design. Wijesundara DK, Xi Y, Ranasinghe C. Front Immunol. 2014 Aug 29;5:412. DOI: 10.3389/fimmu.2014.00412.
It has been well-established that type I interferons (IFN-Is) have pleiotropic effects and play an early central role in the control of many acute viral infections. However, their pleiotropic effects are not always beneficial to the host and in fact several reports suggest that the induction of IFN-Is exacerbate disease outcomes against some bacterial and chronic viral infections. In this brief review, we probe into this mystery and try to develop answers based on past and recent studies evaluating the roles of IFN-Is in infection and immunity as this is vital for developing effective IFN-Is based therapeutics and vaccines. We also discuss the biological roles of an emerging IFN-I, namely IFN-ε, and discuss its potential use as a mucosal therapeutic and/or vaccine adjuvant. Overall, we anticipate the discussions generated in this review will provide new insights for better exploiting the biological functions of IFN-Is in developing efficacious therapeutics and vaccines in the future.

PubChem. Interferon alfa-2B. 2020. National Library of Medicine (US). Maryland: National Library of Medicine (US); c2020.

ClinicalTrials.gov. Maryland: National Library of Medicine (US). Maryland: National Library of Medicine (US); c2020.

MEDIA REPORTS ON CUBAN INTERFERON ALFA-2b

Cuba’s Contribution to Combating COVID-19. Yaffe H. Yale University Press Blog, 2020 Mar 12.
Cuban Interferon Alpha 2b has proven effective for viruses with characteristics similar to those of COVID-19. Cuban biotech specialist Dr. Luis Herrera Martinez explained, “its use prevents aggravation and complications in patients, reaching that stage that ultimately can result in death.” Cuba first developed and used interferons to arrest a deadly outbreak of the dengue virus in 1981, and the experience catalyzed the development of the island’s now world-leading biotech industry.

A history of how Cuba’s Anti-Viral medicine is being used in China. Elizalde RM. RESUMEN LATINOAMERICANO AND THE THIRD WORLD, 2020 Feb 20.
Cuba’s antiviral Recombinant Interferon Alpha 2B (IFNrec) is among the medicines chosen by China to treat the coronavirus, the disease that has already caused at least 1,800 deaths in that country. To date, there is still no specific vaccine.

Cuba Uses ‘Wonder Drug’ to Fight Coronavirus around World Despite U.S. Sanctions. O’Connor T. Newsweek, 2020 Mar 24.
Countries from across the globe have asked Cuba for the recombinant human Interferon Alpha 2B, marketed as Heberon® Alfa R, a drug developed in Cuba proven to be an effective against COVID-19.

Cuban Medication Interferon alpha Selected Among Drugs to Combat Coronavirus.  Periódico Granma, 2020 Feb 10.
Cuban President Miguel Díaz-Canel noted the Chinese government’s determined efforts to confront coronavirus epidemic, with positive results becoming evident, including the recovery of more than 1,500 patients.

El hecho y la noticia del antiviral cubano en China. Elizalde RM. La Jornada, 2020 Feb 20.
La noticia: el antiviral cubano Interferón alfa 2B recombinante (IFNrec) está entre los medicamentos escogidos por China para tratar el coronavirus, la enfermedad que ya ha provocado al menos 1,800 muertos en ese país y para la que todavía no existe una vacuna específica. El hecho: el interferón lleva en Cuba 39 años y el país comenzó el desarrollo de esa proteína con propiedades antivirales en el mismo momento en que se inventaba la industria biotecnológica, en 1981.

How Cuba is leading the world in the fight against coronavirus. McLeod A. MPN News, 2020 Mar 16.
As Cuba sends doctors around the world to help fight coronavirus, a Cuban antiviral drug, Interferon Alpha 2b, is helping countries like China stem the tide of the outbreak.

Promising Treatment for New Human Coronavirus. Collins F. NIH Director’s Blog, 2013 Apr 23.
Quick report that a combination of ribavirin and interferon α2b stops nCoV cells from replicating in the lab.