Recombinant Human Interferon Alpha-2b Research Articles

Production of Recombinant Human Interferon alpha 2b (rhIFN?-2b) in genetically engineered strain of Bacillus subtilis

Interferon alpha 2b (IFN?-2b) is a critical therapeutic protein with immunomodulatory properties, widely used in the treatment of various diseases, including viral infections and cancer. This study presents a comprehensive methodology for the cloning and characterization of the IFN?-2b gene, including the identification of point mutations, followed by expression in Bacillus subtilis. The research begins with gene isolation, primer design, RNA extraction, cDNA synthesis, and PCR amplification. Cloning into a T/A vector and subsequent sequencing reveal point mutations within the gene. The transformed construct is expressed in Bacillus subtilis, and the resulting protein is analyzed using SDS-PAGE. The identification of point mutations and successful protein expression in Bacillus subtilis. This study provides valuable insights into the IFN?-2b gene and its potential applications.

Intralesional interferon alpha-2b as a novel treatment for periocular squamous cell carcinoma in horses.

To determine the safety and efficacy of perilesional human recombinant interferon alpha-2b (IFNα2b) for treatment of periocular squamous cell carcinoma (PSCC) in horses. Eleven horses (12 eyes) with PSCC were enrolled in this prospective clinical study with owner consent. Systemically healthy horses were included in the study following confirmation of PSCC via biopsy. Every two weeks for a maximum of six treatments, horses were sedated and perilesional injection of IFNα2b (10 million IU) was performed. Tumors were measured prior to each injection and at one, three, and 12 months after treatment completion. A greater than 50% reduction in tumor size was considered positive response to treatment (i.e., partial or complete response). Development of anti-IFNα2b antibodies was assessed using serum samples obtained after treatment initiation and compared with treatment responses. Antibody concentrations were analyzed using a mixed model. Statistical significance was considered p < 0.05. Each horse received four to six perilesional injections of IFNα2b. Five of 12 eyes (4/11 horses) responded to treatment. Two of five eyes showed complete resolution of gross PSCC. No systemic adverse effects were seen. Local swelling occurred during treatment protocol in 6/11 horses but resolved without intervention. All horses developed serum anti-IFNα2b antibodies. There was no evidence of statistical difference in antibody concentration between responders and non-responders. Perilesional administration of IFNα2b was found to be well-tolerated in horses with PSCC, and induced tumor regression in 42% of treated eyes. Treatment failure appears unrelated to the development of IFNα2b antibodies.

Engineering a New IFN-ApoA-I Fusion Protein with Low Toxicity and Prolonged Action.

Recombinant human interferon alpha-2b (rIFN) is widely used in antiviral and anticancer immunotherapy. However, the high efficiency of interferon therapy is accompanied by a number of side effects; this problem requires the design of a new class of interferon molecules with reduced cytotoxicity. In this work, IFN was modified via genetic engineering methods by merging it with the blood plasma protein apolipoprotein A-I in order to reduce acute toxicity and improve the pharmacokinetics of IFN. The chimeric protein was obtained via biosynthesis in the yeast P. pastoris. The yield of ryIFN-ApoA-I protein when cultivated on a shaker in flasks was 30 mg/L; protein purification was carried out using reverse-phase chromatography to a purity of 95-97%. The chimeric protein demonstrated complete preservation of the biological activity of IFN in the model of vesicular stomatitis virus and SARS-CoV-2. In addition, the chimeric form had reduced cytotoxicity towards Vero cells and increased cell viability under viral load conditions compared with commercial IFN-a2b preparations. Analysis of the pharmacokinetic profile of ryIFN-ApoA-I after a single subcutaneous injection in mice showed a 1.8-fold increased half-life of the chimeric protein compared with ryIFN.

Cholecalciferol as a means of nonspecific immunoprophylaxis against COVID-19

The current direction of scientific research in recent years has been the study of the immunobiological properties of vitamin D. The purpose of this work was to analyze the results of oral administration of cholecalciferol in order to prevent infection with the SARS-CoV-2 virus in the first wave of the COVID-19 pandemic. The study was performed in the period from October 07 to December 29, 2020, when there were no immunobiological drugs for specific prevention of COVID-19. The total number of respondents was 73 people; all had been ill with coronavirus only once. The etiological diagnosis of the disease included molecular genetic testing of samples of two localizations obtained by the conventional method (nasopharynx, oropharynx). The concentration of antibodies to the virus was determined on average 2 months after the disease using a set of reagents SARS-CoV-2-IgG quantitative-ELISA-Best (JSC Vector-Best, Russia). An approximate assessment of IgM concentration was carried out using a set of SARS-CoV-2-IgM-ELISA-Best from the same manufacturer. Among the study participants were those who used immunobiological drugs for the prevention of infection (riamilovir, umifenovir hydrochloride monohydrate, human recombinant interferon alpha-2b, zinc acetate, vitamin C). In particular, 28 people (38.4%) took cholecalciferol (group No. 1) and 45 people (61.6%) did not use this (group No. 2). Statistical processing of the obtained data was performed using the statistical package STATISTICA v.12.5.192.5 (StatSoft, Inc., USA). We applied the analysis of basic statistics, Linear Discriminant Analysis, Kolmogorov–Smirnov test, Chi-Square test, Wald–Wolfowitz Runs Test, Kruskal– Wallis test. Differences in the incidence of respiratory distress syndrome of the two studied groups were revealed: in patients taking cholecalciferol, the syndrome did not develop at all; in group No. 2, it was registered in 20.0% of cases (Chi-Square = 5.242, p = 0.02). In addition, in patients of group No. 1, the concentration of IgG 2 months after the disease was 3.8 times higher than the values in group No. 2 (Chi-Square = 9.268, p = 0.003). Similar differences were found for the IgM level (Wilks' Lambda: 0.659 approx. F (7.32) = 2.367 p &lt; 0.045). It was known that in both groups there were respondents who used other immuno-active substances for preventive purposes. In the first group there were 18 people (24.7% of all); in the second, there were 13 people (17.8% of all). It was found that those who used other immuno-active substances and did not take vitamin D suffered the disease more easily than everyone else. The respondents who did not use any immunoprophylactic agents were the next in terms of the severity of the infection. The respondents who took cholecalciferol mainly assessed the severity of the infection as average. The study participants who took both vitamin D and used other means of prevention suffered the most from COVID-19. Respondents who took cholecalciferol more often than others reported long-term fatigue, exacerbation of chronic and the appearance of new diseases (hypertension, cardialgia, bronchial asthma, allergies, decreased visual acuity), muscle, joint and vertebral pains that appeared for the first time. The phenomenon of arthralgia and other lesions of large joints in COVID-19 was described by us earlier. Studies by other authors also report frequent complaints of increased fatigue and joint pain. At the same time, the role of vitamin D is considered exclusively from the standpoint of vitamin deficiency in a new coronavirus infection and its potential role in inhibiting hyperinflammatory reactions, as well as accelerating the healing process of affected areas, especially in lung tissue. It was found that vitamin D intake did not affect the incidence of fever, the incidence of pneumonia, the volume of lung tissue damage (based on computed tomography data), the duration of hospitalization and the disease as a whole, and also did not prevent the development of anosmia and dysgeusia. The use of vitamin D as a protective agent to prevent infection with the SARS-CoV-2 virus has had an impact on reducing the frequency/ prevention of cases of respiratory distress syndrome during the disease. Also, those who took vitamin D recorded an increase in the formation of IgG to the SARS-CoV-2 virus 2 months after infection 3.8 times higher than the values recorded in respondents who did not take cholecalciferol. The participants who took cholecalciferol suffered the infection more severely, especially if they used any other protective substances. Also, with the preventive intake of vitamin D after COVID-19, increased fatigue persisted longer, the appearance of new and activation of chronic diseases and muscle, joint and vertebral pains that appeared for the first time were reported more often, which correlates with the data we received earlier.

The effectiveness of Interferon-Ophthalmo, an antiviral and immunotropic drug, in the treatment of adenoviral ophthalmic infections

Adenoviral conjunctivitis (AVC) is treated with pathogenetically determined immunotropic drugs, the most commonly used of which are combined eye drops that include recombinant human interferon alpha-2b and diphenhydramine hydrochloride. In 2020, Russia registered the first domestic generic of the original, called Interferon-Ophthalmo eye drops. Purpose: to compare the effectiveness and safety of InterferonOphthalmo and the original interferon alpha-2b + diphenhydramine drug (Ophthalmoferon) in AVC patients. Material and methods. The study included 30 patients (60 eyes) aged 18–75 years with a clinically confirmed AVC, divided into the main group and the control group, each having 15 patients. The main group received Interferon-Ophthalmo, while the control group received Ophthamoferon. The treatment procedures were identical, with the follow-up period of 15 days. Clinical manifestations of AVC were assessed according to multiple parameters: complaints of eyelid edema, eye redness, lacrimation, itching, foreign body sensation, severity of conjunctival edema and hyperemia, follicular reaction in the lower conjunctival fornix, and hemorrhages. Results. The comparative assessment of the proportion of cases of clinical recovery, performed on the 7th and the 15th days of AVC therapy, confirms the same effectiveness of the two drugs. Conclusion. Interferon-Ophthalmo has a high clinical efficacy comparable with that of the original drug Interferon alpha-2b + Diphenhydramine and can be recommended for the treatment of patients with adenoviral ophthalmic infections.

REBACIN® inhibits E6/E7 oncogenes in clearance of human papillomavirus infection.

http://www.chictr.org.cn/historyversionpuben.aspx?regno=ChiCTR2100045911, identifier ChiCTR2100045911.

Effects of Tenofovir Combined with Recombinant Human Interferon α-2b on Negative Conversion Rate, Liver Function, Immune Status, and Drug Safety in Patients with Chronic Hepatitis B: A Systematic Review and Meta-Analysis.

Objective To systematically evaluate the clinical value of tenofovir combined with recombinant human interferon α-2b in the treatment of chronic hepatitis B and to provide evidence-based medicine for its popularization and use. Methods The randomized controlled trials (RCTs) of tenofovir combined with recombinant human interferon α-2b in the online database of PubMed, EMBASE, ScienceDirect, Cochrane Library, China knowledge Network (CNKI), China VIP database, Wanfang database, and China Biomedical Literature Database (CBM) were searched. The data included in this study were extracted by two independent researchers. After extracting the data of the study, the Cochrane manual 5.1.0 standard was used to evaluate the bias risk of all the literature included in this study. RevMan5.4 statistical software was used to analyze the collected data by meta. Results Entecavir combined with recombinant human interferon α-2b can inhibit the activity of HBV polymerase and improve the inflammatory response of the liver. Recombinant human interferon α-2b can regulate immune function by inducing T cell differentiation and maturation and enhancing the production of cytokines. The systematic evaluation showed that entecavir combined with recombinant human interferon α-2b had higher serum HBeAg negative conversion rate, higher drug safety compared with entecavir alone, and improved liver function and immune status. Conclusion Tenofovir combined with recombinant human interferon alpha-2b has a high serum HBeAg negative rate and safety profile for the treatment of chronic hepatitis B. The combination treatment can improve liver function and immune status in patients, but more studies with higher methodological quality and longer duration of intervention are needed for further validation.

&lt;i&gt;In vitro&lt;/i&gt; antiviral activity of VIFERON® rectal suppositories against SARS-CoV-2

In 2020–2021, the world was engulfed by the pandemic of a new coronavirus infection (COVID-19) caused by the SARS-CoV-2 virus. The low population coverage with vaccination against COVID-19 and the lack of herd immunity result in the need to find an effective and safe etiotropic treatment. Medicinal agents for treatment of COVID-19, approved while preparing this publication, have several limitations related to the conditions of their use and/or population category. In this situation, interferon-containing drugs widely used in Russia and the CIS for prevention and treatment of viral infectious diseases, i.e. ARVI and influenza, may hold promise. This study aims to confirm in vitro antiviral activity against SARS-CoV-2 for the preparation VIFERON® containing recombinant human interferon alpha-2b (IFNα-2b). Materials and methods. Vero CCL-81 cells were infected with hCoV-19/StPetersburg-RII3524VR4/2020 strain of SARS-CoV-2 at doses of 10 TCID50 or 100 TCID50 per well. The suppressive effect of IFN-2b, extracted from VIFERON® in dosage form of rectal suppositories, was evaluated by qRT-PCR at 24 h and 48 h after the infection of cells in two schemes, simulating preventive (24 h before infection) and therapeutic (2 h after infection) use of drugs. Results. IFNα-2b at concentrations of 800, 400, 200, 100 and 50 IU/ml, extracted from rectal suppositories of VIFERON®, showed high biological activity, displayed as inhibition of SARS-CoV-2 strain replication in both infectious doses evaluated either at 24 h or at 48 h after cell infection. The “preventive” vs. “therapeutic” scheme was found to be more effective. In the “preventive” scheme the virus titre decreased by more than 3 lg TCID50 at 24 hours post-infection and by 5–6 lg TCID50 at 48 hours post-infection after administration of 800 IU/ml IFNα-2b. Conclusion. The study results evidence that VIFERON® in dosage form of rectal suppositories may be promising for prevention and treatment of new coronavirus infection in clinical practice.

Anti-Lethal Therapy of COVID-19 for Home Health Outpatient Therapy

The article presents the rationale for a new therapeutic strategy for treatment of patients with COVID-19, based on the combined use of biologic response modifiers and drugs targeting etiology and pathophysiology of the novel coronavirus infection, excluding disadvantages of polypharmacy, and providing a high clinical effect. This approach has been predominantly used in home health outpatient treatment of 324 patients with COVID-19 of variable severity using the biologic response modifiers “double drug cocktail” made of recombinant human interleukin-2 and recombinant human interferon alpha-2b in combination with alimemazine, nimesulide, rivaroxaban and antibiotic therapy (co-amoxiclav, or ceftriaxone) if secondary bacterial pneumonia was diagnosed. The results obtained indicate a dramatic improvement in the condition of patients, even with a severe COVID-19, which made possible to avoid artificial ventilation and prevent deaths.

Novel drug for COVID-19: Virafin a therapeutic breakthrough

Background: Pegylated Interferon Alpha-2b is synthesized via the use of PEG (polyethylene glycol). It is formulated via attaching a 12kDa mono methoxy polyethylene glycol moiety to the recombinant human Interferon Alpha-2b protein. Pegylated Interferon Alpha-2b acts via binding to the (JAK/STAT) Janus kinase signal transducer and activator of the transcription interferon receptor. The Pegylated Interferon Alpha-2b /Virafin binds to the (IFNAR1/2) alpha interferon receptor 1 and 2. Pegylated Interferon Alpha-2b better known as Virafin has been approved by the Drugs Controller General of India. This therapeutic agent is currently under a multicentric phase 3 trial with very promising results being reported. A single subcutaneous dose of Virafin has been shown to decrease the need for oxygen therapy in patients. This reduction in the need for oxygen therapy is a vital factor needed to provide relief to the Indian medical system in light of the recent oxygen shortages faced due to India’s worst wave of COVID-19 cases since the onset of the global pandemic. Pegylated Interferon Alpha-2b/ Virafin confers enhanced viral clearance and bolsters the immune response to induce a quicker recovery in patients with mild to moderate symptoms.&#x0D; Conclusion: It is of paramount importance that further research on Virafin is undertaken as it can hinder the progression of COVID-19, reduce pressure on the inundated health systems, and save countless lives.

Применение препарата интерферона альфа-2b в комплексном лечении пациентов с COVID-19

Objective. To evaluate the efficacy and safety of human recombinant interferon alpha-2b (VIFERON®) as a part of comprehensive treatment for coronavirus infection COVID-19. Patients and methods. This prospective, comparative, controlled study included 140 patients with COVID-19 randomized in two groups. The experimental group (EG) included 70 patients who received standard therapy plus VIFERON® (one rectal suppository 3,000,000 IU three times a day and gel 36,000 IU/g 5 times a day applied to the nasal mucosa and palatine tonsils for 14 days); the control group (CG) comprised 70 patients who received standard therapy alone. Results. Patients in the EG demonstrated more rapid resolution of main symptoms, such as intoxication, bronchopulmonary and catarrhal manifestations. Normalization of the total score in the EG was observed 7 days earlier than in the CG. In the EG, the proportions of patients who had their D-dimer and CRP levels normalized were 42.7% and 18.7% higher than those in the CG, respectively (р &lt; 0.05). Follow-up computed tomography demonstrated that the proportion of patients with positive dynamics in the EG was 8.1% higher than that among controls, whereas advanced disease with 51%–75% of lung tissue affected was 11.9% less common in participants from the EG than in controls (р = 0.019). We observed no adverse events associated with interferon alpha-2b (VIFERON®) or other medicines included in the treatment scheme. Conclusion. Our findings suggest high efficacy and safety of recombinant interferon alpha-2b (VIFERON®) used in combination with symptomatic agents, antibiotics, anticoagulants, which allows us to recommend this drug for inclusion into standard treatment schemes for COVID-19. Key words: COVID-19, coronavirus infection, human recombinant interferon alpha-2b

Эффективность рекомбинантного интерферона альфа-2b при интраназальном применении для экстренной профилактики COVID-19 у медицинских работников

Healthcare workers are one of the main risk groups for COVID-19 infection and should be provided priority protection. Objective. To examine the development dynamics and epidemic process manifestations of COVID-19 in healthcare workers as well as to assess the efficacy of intranasal human recombinant interferon alpha-2b (Grippferon®) for emergency medicationassisted nonspecific prevention of novel coronavirus infection. Patients and methods. The incidence of COVID-19 in Perm healthcare workers was examined using official statistical data in the epidemic dynamics. The prophylactic efficacy of Grippferon® was assessed in the analytic cohort prospective study, involving 561 healthcare workers of infectious diseases hospital departments. Results. The incidence rate of COVID-19 among healthcare workers in Perm for the examined period from 09.03.2020 to 06.12.2020 was four times higher, than in the general population. Healthcare workers aged 30–59 years were found to be more likely to develop COVID-19. Physicians had the highest risk of infection compared to other medical professionals. The prophylactic efficacy of Grippferon® was shown to be high in physicians and nurses, who were in contact with COVID-19 patients. Conclusion. Recombinant interferon alpha-2b-based medication Grippferon® can be recommended for emergency prevention of novel coronavirus infection to ensure epidemiological safety of healthcare workers and patients during the COVID-19 pandemic. Key words: COVID-19, incidence rate, healthcare workers, recombinant interferon alpha-2b, Grippferon, intranasal use, prevention

Features of the course and therapy of HIV infection in children at different stages of the disease

Purpose of the study . Analysis of clinical and laboratory features of the course of HIV infection and antiviral therapy in children at different stages of the disease. Materials and methods. Under supervision in the department of motherhood and childhood there were 90 children with HIV infection, which were divided into 3 groups: 1st group – 35 people. with a diagnosis of HIV infection in the first 2 months of life, group 2 – 25 people. with a diagnosis at the age of 6-12 months and the 3rd group (30 people) with a diagnosis of HIV infection over the age of 2 years. Statistical processing of the research results was carried out using the software package Statistica for Windows (version 8.0). Research results. Early detection of HIV infection and the beginning of antiretroviral therapy (ARVT) led to the absence of clinical manifestations of the disease, low levels of HIV RNA in the blood and normal levels of CD4 lymphocytes, which persisted for 3 years of observation. The detection of HIV infection and the start of ART in the first 6–12 months of life of children was characterized by clinical symptoms up to CNS damage, high levels of HIV RNA in the blood, moderate decrease in CD4 lymphocytes, with normalization after 3–6 months of treatment and after 3 years of observation. Late diagnosis of HIV infection in children – over 2 years of age is characterized by severe forms of the disease with secondary manifestations – AIDS in 20,0% of cases, low rates of CD4 lymphocytes, damage to the central nervous system with the formation of spastic diplegia, and slow recovery of immunity. The use of the drug Viferon, rectal suppositories in the combined treatment of EBV-mononucleosis and a generalized form of CMV infection allows the early treatment of the main symptoms of herpes virus infections and normalization of laboratory parameters. Conclusion Our prospective study showed that mother-tochild transmission of HIV was realized due to the absence of the first stage of chemoprophylaxis (during pregnancy), and was also aggravated in 9 cases (10,0%) by breastfeeding. The severity of the clinical manifestations of HIV infection, the levels of CD4-lymphocytes and HIV RNA in the blood directly depend on the period of detection and the initiation of ART in children. The inclusion of human recombinant interferon alpha-2b with antioxidants (Viferon rectal suppositories) in the etiopathogenetic therapy of comorbid herpes virus diseases (EBV-mononucleosis, a generalized form of CMV infection) is effective and safe.

Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity

Specific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most appropriate conditions for in vitro determination of IFN-α product specific activity. Materials and methods: Vero, MDBK, Hep-2, and A-549 homologous and heterologous cell cultures, as well as vesicular stomatitis Indiana virus (VSV) and murine encephalomyocarditis (EMC) virus at a dose of 100 TCD50 /0.1 mL were used for determination of specific antiviral activity. The international reference standard of recombinant interferon alpha-2b activity (Interferon alpha 2b, human, rDNA, E. coli-derived, 2nd WHO International Standard, 1999, NIBSC Code No. 95/566) and human recombinant interferon alpha 2b in the form of solution (batch No. 040214, Pharmapark LLC, Russia) were used as IFN-α samples. Results: the analysis of the obtained data helped to determine: the combinations of cell lines and the indicator virus most sensitive to IFN-α; the optimal concentration of fetal serum in the medium, and the optimal time parameters; the preferred method of reporting test results. Conclusions: the following test conditions were found to be optimal: the MDBK/VSV and Hep-2/EMC combinations proved to be the most sensitive to IFN-α; the optimal period of interferon and cell culture incubation—24 hours; the optimal concentration of fetal bovine serum in the culture medium used for diluting interferon products—2–5%. The instrumental procedure is preferred for reporting the results of interferon antiviral activity determination, because it is up-to-date, reliable, accurate and time-efficient.

A prospective multicenter randomized controlled study on the efficacy and safety of pharyngeal spraying recombinant human interferon alpha 2b in the treatment of children with herpangina

Objective To evaluate the efficacy and safety of pharyngeal spraying recombinant human interfe-ron alpha 2b (rhIFNα2b) in the treatment of herpangina in children. Method s A prospective, multicenter, rando-mized, opened and controlled study was carried out in 11 hospitals in Anhui province from August 2018 to March 2019.According to the time of admission, 180 patients diagnosed as herpangina were prospectively and randomly divided into rhIFNα2b treatment group and Ribavirin control group.On the basis of giving both groups the heat-clearing, detoxifying and anti-infection treatment, the patients in treatment group received pharyngeal spraying rhIFNα2b 9 g/L saline solution[1 million IU/mL, 0.1 million IU/(0.1 mL·press)], and the patients in control group were treated by pharyngeal spraying Ribavirin (0.5 mg RBV/press, 150 press), 3 presses per time, 4 times per day, continuous administration for 5 days for both groups.Those who recovered in advance were no longer given medication.All patients were observed to fully recover.The clinical efficacy and the disappearing time of symptoms and signs between two groups were compared, and the safety of pharyngeal spraying rhIFNα2b for patients was evaluated. Result s All of the 180 patients completed the study, including 90 cases in the treatment group and 90 cases in the control group.There was no statistically significant difference in terms of gender, age, weight and course of illness before treatment between the two groups (all P>0.05), which had clinical comparability.The apparent efficiency of the treatment group [63.3% (57/90 cases)] was significantly higher than that in the control group [38.9% (35/90 cases)] and the difference was statistically significant(χ2=10.934, P=0.004); no significant difference in the total efficiency between the treatment group [96.7% (87/90 cases)]and the control group [92.2% (83/90 cases)]was observed (χ2=2.924, P=0.169). The duration of fever[(32.59±20.73) h vs.(45.72±26.96) h], hyperemia[(76.48±23.12) h vs.(92.44±24.31) h], herpes[(72.99±25.77) h vs.(85.09± 26.62) h], salivation[(45.44±24.96) h vs.(54.42±31.20) h] and anorexia[(62.70±23.99) h vs.(78.71±30.54) h] in the treatment group were significantly shorter than those in the control group, and the differences were statistically significant(all P 0.05). After treatment, the serum levels of TNF-α and IL-6 were(7.26±1.99) ng/L and (2.42±0.73) ng/L in the treatment group, which were significantly lower than those in the control group [(12.09±6.39) ng/L and (7.32±11.51) ng/L](all P 0.05). The comparison on positive rate of virus in pharyngeal swab between the treatment group [65.3% (32/49 cases) and 40.6% (13/32 cases) respectively] and the control group[66.7%(36/54 cases) and 41.0% (16/39 cases), respectively]before and after therapy showed no significant difference (all P>0.05). During the treatment, no serious adverse reactions were observed in the two groups.The incidence of adverse reactions was 1.1% (1/90 cases) in the treatment group and 5.6% (5/90 cases) in the control group.In addition, the serum hemoglobin level of children in the control group after treatment was significantly lower than that before treatment and that in the treatment group (all P<0.05). Conclusions Compared with pharyngeal spraying ribavirin, pharyngeal spraying rhIFNα2b can greatly improve the clinical efficiency, accelerate the disappearance of clinical symptoms and signs, and shorten the total course of disease, and is more safe and worthy of clinical application. Key words: Herpangina; Recombinant human interferon alpha 2b; Ribavirin; Pharyngeal spraying

Лечение урогенитальных инфекций у беременных женщин и внутриутробного инфицирования у новорожденных детей препаратами рекомбинантного интерферона альфа-2b: результаты мета-анализа

We summarized the results of 35 studies published in the last 25 years that evaluated the efficacy and safety of human recombinant interferon alpha-2b (Viferon®) in the treatment of inflammatory genitourinary infections in pregnant women and intrauterine infections in newborns. This drug has a positive effect on clinical manifestations and disease outcome; it promotes the elimination of pathogens, reduces the frequency and duration of viral/bacterial shedding and recurrent infections; it also prevents coinfection with other pathogens; reduces infection-associated morphological changes of the placenta and improves its morphometric parameters; restores normal uteroplacental blood flow, reduces the risk of placental insufficiency and intrauterine hypoxia; it significantly changes the colposcopy pattern, restore vaginal microbiocenosis, substantially reduces the incidence of infectious and inflammatory complications in mothers and incidence of intrauterine infections (including severe ones) in newborns; it decreases the length of stay in hospital and intensive care unit, duration of artificial ventilation, parenteral nutrition, antibacterial therapy in newborns with pneumonia; significantly reduces the frequency of perinatal non-infectious disorders; it improves the immune parameters, including those associated with antiviral protection, as well as the cytokine status in pregnant women with genitourinary infections and newborns with intrauterine infections; the drug caused no side effects or adverse reactions. Key words: pregnant women, intrauterine infection, newborns, recombinant interferon alpha-2b, urogenital infections

Safety of aerosol inhalation of recombinant human interferon alpha 2b injection(P.putida)and excipient hydroxyethyl starch 40 to healthy SD rats

Objective To investigate the effects of recombinant human interferon alpha 2b injection(P.putida)and hydroxyethyl starch inhalation on physiological indexes and tissues and organs of SD rats, such as lungs, trachea and bronchus, and thus to explore the safety of the drug and excipients for inhalation. Methods Rats were randomly divided into two groups(hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group).5 million IU/ml recombinant human interferon alpha 2b injection or 6% hydroxyethyl starch 40 injection were given respectively, which were diluted with 0.9% sodium chloride injection in a radio of 1: 2.Both groups were given aerosol inhalation for 30 minutes once a day for 14 days.Within 14 days after administration, the behavior of rats and the degree of toxicity were continuously observed.During the drug administration and recovery period, the clinical signs of the rats in each group were observed daily, and the body weight and food consumption were measured once a week.At the end of drug administration and the end of recovery period, hematology test, blood biochemical test, gross anatomical observation, organ weight determination, and histopathological examination were performed. Results The mass median aerodynamic diameter(MMAD)of the drug mist was 3.56 μm, and the geometric standard deviation(GSD)was 1.84 μm, which met the particle size requirements of arrival and deposition in the lower respiratory tract via atomization inhalation.No abnormality in clinical signs, body weight and food consumption, hematology and blood biochemical indexes were found in hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group.No abnormal histopathological changes were observed in oral mucosa, tongue, nasal cavity(paranasal sinus), larynx, trachea, main bronchial tube, lung, heart, liver, spleen, kidney, reproductive system and other organs or tissues examined. Conclusion Results suggest the drug particle size of recombinant human interferon alpha 2b injection(pseudomonas)can reach the lower respiratory tract and deposit in the lower respiratory tract.Continuous atomization inhalation for 14 days has a good safety, and the excipient hydroxyethyl starch 40 has a high safety. Key words: Recombinant human interferon alpha 2b; Injection; Aerosol inhalation; Hydroxyethyl starch 40; Safety

Vaginal biocenosis in pregnant women with recurrent bacterial vaginosis after treatment with recombinant human interferon alpha-2b

Introduction.Preventing recurrent bacterial vaginosis (BV) during pregnancy is an important issue in current obstetrics.Aim:to assess the effect of recombinant human interferon alpha-2b on the incidence of recurrent BV during pregnancy and on the bacterial, cytokine, and antimicrobial components of the nonspecific vaginal immunity.Materials and methods.The study included 80 pregnant women with recurrent BV, 40 of whom (the main group) were treated with recombinant human interferon alpha-2b in the first half of pregnancy and after 30 weeks of pregnancy in addition to standard therapy (metronidazole or clindamycin). The other 40 women (control group) did not receive this interferon drug. Patients of the both groups were examined for the rate of BV recurrence and also tested for their vaginal bacteria and levels of β-defensin-2 (HBD-2), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and interferon-γ (INF-γ) in the third trimester of pregnancy.Results.Treatment with recombinant human interferon alfa-2b led to a 3-fold reduction in the occurrence of repeated episodes of BV and an increase in the content of lactobacilli (7.24 ± 0.33 and 7.03 ± 0.40 in patients with interferon therapy and without it, respectively; p = 0.013) mainly due to Lactobacillus crispatus (7.14 ± 0.69 in women who received interferon alfa-2b, compared with 6.36 ± 0.95 in patients on standard therapy; p = 0.003) and a decrease in the number of some types of anaerobes. Also, the use of interferon alpha-2b in pregnant women with recurrent BV led to an increase in the content of IL-1β by 2.5 times, IL-6 by 3.5 times, IL-10 by 1.5 times, INF-γ by 2 times, and HBD-2 by 4 times.Conclusion.Treatment of recurrent BV during pregnancy with recombinant human interferon alfa-2b, aimed at boosting the nonspecific vaginal immunity helps reduce the incidence of recurrent BV.

High level extracellular production of recombinant human interferon alpha 2b in glycoengineered Pichia pastoris: culture medium optimization, high cell density cultivation and biological characterization.

The present study was aimed at design of experiments (DoE)- and artificial intelligence-based culture medium optimization for high level extracellular production of a novel recombinant human interferon alpha 2b (huIFNα2b) in glycoengineered Pichia pastoris and its characterization. The artificial neural network-genetic algorithm model exhibited improved huIFNα2b production and better predictability compared to response surface methodology. The optimized medium exhibited a fivefold increase in huIFNα2b titre compared to the complex medium. A maximum titre of huIFNα2b (436mgl-1 ) was achieved using the optimized medium in the bioreactor. Real-time capacitance data from dielectric spectroscopy were utilized to model the growth kinetics with unstructured models. Biological characterization by antiproliferative assay proved that the purified recombinant huIFNα2b was biologically active, exhibiting growth inhibition on breast cancer cell line. Culture medium optimization resulted in enhanced production of huIFNα2b in glycoengineered P. pastoris at both shake flask and bioreactor level. The purified huIFNα2b was found to be N-glycosylated and biologically active. DoE-based medium optimization strategy significantly improved huIFNα2b production. The antiproliferative activity of huIFNα2b substantiates its potential scope for application in cancer therapy.

Molecular and Biological Characterization of a Prepared Recombinant Human Interferon Alpha 2b Isoform.

Recombinant human interferon alpha2b (rhIFN-α2b) protein is FDA approved for treatment of many tumors and viral diseases. A rhIFN-α2b isoform has been produced and purified from the refolding reaction using high-resolution anion ion exchange chromatography. This isoform has a proper MW (19kDa) and high purity and homogeneity. The conservation of native linear and conformational epitopes in this isoform was immunologically confirmed by Western blot and ELISA. Mass spectrometry assessment of its intact mass showed average mass (19,337Da) equivalent to that of the expressed rhIFN-α2b protein without any chemical modification and without the first methionine. Peptide mapping of rhIFN-α2b through tryptic digestion of reductive/alkylated protein using urea as a denaturing agent gave the best pattern. The rhIFN-α2b had a high specific antiviral activity (2.5 × 108 ± 1.1 × 108IU/mg protein). In vivo clearance study of rhIFN-α2b in female SD rats (500μg/kg, intramuscularly) revealed rapid clearance (elimination half-life 0.54h with a maximum plasma concentration of 33,792pg/ml) compared with the commercial rhIFN-α2 (elimination half-life 0.75-0.96h). In conclusion, the prepared rhIFN-α2b isoform has high purity, homogeneity, native like chemical and structural composition, high antiviral activity, and proper biological stability, which reduce its immunogenicity and raise its therapeutic efficiency.