Influence of traditional Chinese medicines on the in vivo metabolism of lopinavir/ritonavir based on UHPLC-MS/MS analysis / 药物分析学报

A fast,reliable,and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019,namely,Lianhua Qingwen granules,Huoxiang Zhengqi capsules,Jinhua Qinggan granules,Shufeng Jiedu capsules,and Angong Niuhuang pills,on the pharmacokinetics of lopinavir/ritonavir in rats.Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard(IS).Separation was performed on an Agilent ZORBAX eclipse plus C18(2.1 mm x 50 mm,1.8 μm)column using acetonitrile and water containing 0.1％formic acid as the mobile phase for gradient elution.The flow rate was 0.4 mL/min and the injection volume was 2 μL Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir,m/z 721.3→296.1 for rito-navir,and m/z 705.4→168.1 for the IS.The method showed good linearity in the concentration range of 25-2500 ng/mL(r=0.9981)for lopinavir and 5-500 ng/mL(r=0.9984)for ritonavir.The intra-day and inter-day precision and accuracy were both within±15％.Items,such as dilution reliability and residual effect,were also within the acceptable limits.The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats.The pharmaco-kinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approx-imately 1.5-to 2-fold relative to that in the control group.Similarly,the pharmacokinetic parameters of lopinavir were altered.Overall,the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.

Toxicodermia as an adverse event in a covid-19 patient: case report

ABSTRACT: Introduction: SARS-COV2 infection, which was initially associated with respiratory manifestations only, can also cause gastrointestinal, kidney, neurological and cardiovascular symptoms according to some reports. Case presentation: A 36-year-old female patient attended the emergency department due to dyspnea, asthenia, adynamia, mild odynophagia. and headache. The patient's medical history included obesity, smoking, and working as a health care worker. Considering the symptoms, antimalarial and antiretroviral treatment was indicated to treat COVID-19, a diagnosis that was confirmed three days after admission. However, on the fourth day of treatment, the patient presented with polydipsia and macular, pruritic, and generalized rash. Due to suspicion of toxicoderma, the treatment was suspended, and the skin condition improved. After 8 days of hospitalization, the patient was discharged with biosecurity recommendations and mandatory isolation for 28 days. Conclusion: The described case is a report of toxicoderma in a patient with COVID-19 under treatment with antiretroviral and antimalarial drugs. Based on the findings, a thorough examination of skin and mucosa of patients with suspected or confirmed COVID-19 will undoubtedly contribute to the correct characterization of this new disease.

RESUMEN Introducción. La infección por SARS-COV2, que en principio se pensó solo causaba manifestaciones respiratorias, también puede ocasionar síntomas gastrointestinales, renales, neurológicos, cardiovasculares e incluso cutáneos según algunos reportes. Presentación del caso. Paciente femenina de 36 años quien asistió al servicio de urgencias por cuadró clínico consistente en disnea, astenia, adinamia, odinofagia leve y cefalea. Como antecedentes de relevancia se registró obesidad, tabaquismo y ocupación como trabajadora de la salud. Dados los síntomas, se indicó tratamiento antimalárico y antirretroviral para tratar COVID-19, diagnóstico que fue confirmado a los tres días de ingreso, pero al cuarto día de instaurado este manejo la mujer presentó polidipsia y rash macular, pruriginoso y generalizado. Por sospecha de toxicodermia, el tratamiento fue suspendido y con esto el cuadro cutáneo mejoró. Luego de 8 días de hospitalización, la paciente recibió el alta, junto con recomendaciones de bioseguridad y confinamiento durante 28 días. Conclusiones. El caso descrito corresponde a un evento de toxicodermia en una paciente con COVID-19 en manejo con antirretroviral y antimalárico. A partir de los hallazgos, se establece que la exploración minuciosa de piel y mucosas en los pacientes con sospecha o diagnóstico confirmado de COVID-19 puede ser de gran ayuda para la correcta caracterización de esta nueva enfermedad.

Smartphone electrocardiogram for QT interval monitoring in Coronavirus Disease 2019 (COVID-19) patients treated with Hydroxychloroquine

@#Introduction: The global pandemic of Corona Virus Disease 2019 (COVID-19) has led to the re-purposing of medications, such as hydroxychloroquine and lopinavir-ritonavir in the treatment of the earlier phase of COVID-19 before the recognized benefit of steroids and antiviral. We aim to explore the corrected QT (QTc) interval and ‘torsadogenic’ potential of hydroxychloroquine and lopinavir-ritonavir utilising a combination of smartphone electrocardiogram and 12-lead electrocardiogram monitoring. Materials and Methods: Between 16-April-2020 to 30-April2020, patients with suspected or confirmed for COVID-19 indicated for in-patient treatment with hydroxychloroquine with or without lopinavir-ritonavir to the Sarawak General Hospital were monitored with KardiaMobile smartphone electrocardiogram (AliveCor®, Mountain View, CA) or standard 12-lead electrocardiogram. The baseline and serial QTc intervals were monitored till the last dose of medications or until the normalization of the QTc interval. Results: Thirty patients were treated with hydroxychloroquine, and 20 (66.7%) patients received a combination of hydroxychloroquine and lopinavir-ritonavir therapy. The maximum QTc interval was significantly prolonged compared to baseline (434.6±28.2msec vs. 458.6±47.1msec, p=0.001). The maximum QTc interval (456.1±45.7msec vs. 464.6±45.2msec, p=0.635) and the delta QTc (32.6±38.5msec vs. 26.3±35.8msec, p=0.658) were not significantly different between patients on hydroxychloroquine or a combination of hydroxychloroquine and lopinavir-ritonavir. Five (16.7%) patients had QTc of 500msec or more. Four (13.3%) patients required discontinuation of hydroxychloroquine and 3 (10.0%) patients required discontinuation of lopinavirritonavir due to QTc prolongation. However, no torsade de pointes was observed. Conclusions: QTc monitoring using smartphone electrocardiogram was feasible in COVID-19 patients treated with hydroxychloroquine with or without lopinavir-ritonavir. The usage of hydroxychloroquine and lopinavir-ritonavir resulted in QTc prolongation, but no torsade de pointes or arrhythmogenic death was observed.

Lopinavir/ritonavir: is early administration better in Covid-19?

Background: To share the data of coronavirus 2019 (Covid-19) patients started on lopinavir-ritonavir (lopi/r) in relation to time period from the onset of symptoms.Method: Observational descriptive study of 23 Covid-19 patients admitted in a tertiary care center in India from March 2020 to May 2020. Patients categorized into 2 groups based on the timing of initiation of lopi/r from the onset of symptoms. Group 1 were given the drug early (≤7 days) and group 2 late (>7 days). The clinical events (oxygen requirement days and ICU stay) and outcomes of hospital stay between the two groups were evaluated.Results: Patients were started on lopi/r for a period of 14 days on admission, out of which 12 patients were in group 1 and 11 patients in group 2. Underlying co-morbidities were present in 15 patients (65.21%). The mean duration from onset of symptoms to lopi/r initiation was 4 days and 11.1 days in Group 1 and 2 respectively. Requirement for oxygen support (2.16 versus 6.54 days), mean duration of hospitalization (8.58 versus 11.54 days) and mean duration of obtaining first Covid-19 negative report from the onset of symptoms (10.5 versus 19.57 days) were all significantly lesser in group 1 (p<0.05). All patients belonging to Group 1 and eight patients of group 2 recovered completely and were discharged whereas 3 patients of group 2 expired. Diarrhea was the most commonly observed adverse effect of lopi/r in our patients.Conclusion: With no approved weapon to tackle the Covid-19 pandemic, we should keep lopi/r in our armamentarium of drugs and use it at the earliest. More clinical trials are needed in future to ascertain if lopi/r can reduce hospital stay, prompt faster recovery and result in better clinical outcome.

Efectos adversos de lopinavir/ritonavir en enfermedad grave por coronavirus (COVID-19) / Adverse effects of lopinavir/ritonavir in critically ill patients with COVID-19

Resumen Durante el transcurso de la pandemia causada por el virus SARS-CoV-2 se han utilizado diferentes fármacos como potenciales tratamientos específicos con el objetivo de lograr mejoría clínica y/o disminuir la mortalidad de los afectados, pero al tratarse de una enfermedad hasta ahora desconocida, la evidencia acerca de su seguridad y eficacia se va construyendo a medida que se los prescribe. La farmacovigilancia intensiva en este contexto permite detectar eventos adversos y mediante su reporte y análisis inferir el perfil de seguridad en cada indicación. Se realizó un estudio observacional, retrospectivo, en un único centro, en el cual se relevaron los eventos adversos en 23 pacientes adultos en estado crítico, de los cuales 18 recibieron lopinavir/ ritonavir como tratamiento empírico, entre el 15 de marzo y el 15 de junio de 2020, durante su internación en una Unidad de Cuidados Intensivos. Se describe el tipo de eventos adversos, su gravedad y si fueron motivo de suspensión del tratamiento. Los resultados del presente análisis muestran una alta tasa de eventos adversos (10/23, 43%) entre los que recibieron lopinavir/ritonavir, llevando en la mayoría de los casos a la decisión de suspender el mismo antes de completar el tratamiento. Aun con las limitaciones propias del reducido número de casos, la divulgación de dichos resultados aporta evidencia para definir el perfil de seguridad de la combinación lopinavir / ritonavir usado en enfermedad grave por SARS-CoV-2.

Abstract During the SARS-CoV-2 pandemic many drugs have been used as potential treatments in order to improve the clinical outcome and reduce the mortality. But since it is a currently unknown disease, the evidence about efficacy and safety is built as the drugs are prescribed. In this context, intensive pharmacovigilance allows early detection of adverse events, and thereby infer the safety profile of the indication. We conducted an observational, retrospective, single-center study involving adult patients with severe SARS-CoV-2 infection. All adverse events detected in 23 patients in the Intensive Care Unit between March 15 and June 15, 2020 were registered. We describe type and severity of the adverse events and if treatment suspension was needed. The results show a high rate of adverse events (10/23, 43%) in treatment with lopinavir/ritonavir. In most cases early treatment suspension was required. Even though the limitations of our study derived from the small sample size, these results could help in building evidence about the safety of using lopinavir/ritonavir for severe SARS-CoV-2 infection.

Lopinavir/ritonavir for COVID-19: a living systematic review / Lopinavir/ritonavir para COVID-19: una revisión sistemática viva

Objective: Provide a timely, rigorous, and continuously updated summary of the evidence on the role of lopinavir/ritonavir in the treatment of patients with COVID-19. Methods: We conducted searches in the special L·OVE (Living OVerview of Evidence) platform for COVID-19, a system that performs regular searches in PubMed, Embase, CENTRAL, and other 33 sources. We searched for randomized trials and non-randomized studies evaluating the effect of lopinavir/ritonavir versus placebo or no treatment in patients with COVID-19. Two reviewers independently evaluated potentially eligible studies, according to predefined selection criteria, and extracted data using a predesigned standardized form. We performed meta-analyses using random-effect models and assessed overall certainty in evidence using the GRADE approach. A living, web-based version of this review will be openly available during the COVID-19 pandemic. Results: Our search strategy yielded 862 references. Finally, we identified 12 studies, including two randomized trials, evaluating lopinavir/ritonavir, in addition to standard care versus standard care alone in 250 adult inpatients with COVID-19. The evidence from randomized trials shows lopinavir/ritonavir may reduce mortality (relative risk: 0.77; 95% confidence interval: 0.45 to 1.3; low certainty evidence), but the anticipated magnitude of the absolute reduction in mortality, varies across different risk groups. Lopinavir/ritonavir also had a slight reduction in the risk of requiring invasive mechanical ventilation, developing respiratory failure, or acute respiratory distress syndrome. However, it did not lead to any difference in the duration of hospitalization and may lead to an increase in the number of total adverse effects. The overall certainty of the evidence was low or very low. Conclusions: For severe and critical patients with COVID-19, lopinavir/ritonavir might play a role in improving outcomes, but the available evidence is still limited. A substantial number of ongoing studies should provide valuable evidence to inform researchers and decision-makers soon.

Objetivo: Esta revisión sistemática viva tiene como objetivo entregar un resumen oportuno, riguroso y constantemente actualizado de la evidencia disponible sobre los efectos de lopinavir/ritonavir en pacientes con COVID-19. Métodos: Se realizó una búsqueda en la plataforma L·OVE COVID-19 (Living OVerview of Evidence), un sistema que mantiene búsquedas regulares en PubMed/Medline, Embase, Cochrane Central Register of Controlled Trials (CENTRAL) y otras 33 fuentes. Se buscaron ensayos aleatorios y estudios no aleatorios que evaluaran el uso de lopinavir/ritonavir versus placebo o ningún tratamiento en pacientes con COVID-19. Dos revisores evaluaron de forma independiente los artículos potencialmente elegibles, de acuerdo con criterios de selección predefinidos, y extrajeron los datos mediante un formulario estandarizado. Los resultados fueron combinados mediante un metanálisis utilizando modelos de efectos aleatorios y evaluamos la certeza de la evidencia utilizando el método GRADE. Una versión viva de esta revisión estará disponible durante la pandemia de COVID-19. Resultados: La búsqueda inicial arrojó 862 referencias. Finalmente, identificamos 12 estudios incluyendo 2 ensayos aleatorios, que evaluaban lopinavir/ritonavir adicionado al tratamiento estándar versus tratamiento estándar en 250 pacientes adultos hospitalizados con COVID-19. Los resultados provenientes de los ensayos aleatorios muestran que el uso de lopinavir/ritonavir puede reducir la mortalidad (riesgo relativo: 0,77; intervalo de confianza 95%: 0,45 a 1,3; certeza de evidencia baja), pero la magnitud de la reducción absoluta de la mortalidad varía según los diferentes grupos de riesgo. El uso de lopinavir/ritonavir mostró además una ligera reducción en el riesgo de requerir ventilación mecánica invasiva, desarrollar insuficiencia respiratoria o síndrome de dificultad respiratoria aguda. No se observó diferencias en la duración de la hospitalización y su uso puede producir un aumento en el número de efectos adversos totales. La certeza global de la evidencia fue baja o muy baja. Conclusiones: Para pacientes graves y críticos con COVID-19, el uso de lopinavir/ritonavir podría desempeñar un papel en la mejora de los resultados, pero la evidencia disponible aún es limitada. La gran cantidad de estudios en curso deberían proporcionar evidencia valiosa para informar a los investigadores y los tomadores de decisiones en el futuro cercano.

Repurposing antivirals as potential treatments for SARS-CoV-2: From SARS to COVID-19

The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the agent of the current pandemic of therespiratory disease known as coronavirus disease 2019 (COVID-19). The government and health authorities aroundthe world have advocated social distancing, containment measures, and effective diagnosis as the first measuresto slow down the spread of the disease, but, still, treatment options are urgent, especially for patients evolving tosevere pneumonia. Several pharmaceuticals with antiviral effects were identified and tested, to some extent, duringthe previous SARS-CoV and Middle East respiratory syndrome coronavirus outbreaks. Type I interferons (IFNs),ribavirin, lopinavir/ritonavir, chloroquine/hydroxychloroquine, and remdesivir emerge as the primary options forin-hospital treatment of patients with COVID-19, focused on reducing the viral load. Although more experimental andclinical evidence is required, the accumulated in vitro and clinical knowledge discussed here supports those drugs asfeasible alternatives to face the SARS-CoV infection in the short term, whereas more effective measures arise fromthe world scientific community

Posibles estrategias de tratamiento para COVID-19 / Possible treatment strategies for COVID-19

No hay tratamiento antiviral específico para el COVID-19. Sin embargo, conocimientos adquiridos durante los brotes del SARS y el MERS, en conjunto con la información obtenida con COVID-19, han permitido detectar varios objetivos terapéuticos en el ciclo de replicación del virus, y en su patogénesis. Se incluye la evidencia actual con respecto a los principales tratamientos propuestos para COVID-19, reutilizados o experimentales, mediante una revisión de la literatura científica a la fecha. Debido a la falta de ensayos controlados aleatorios, se incluyeron: informes de casos, series de casos y artículos de revisión. Globalmente se están llevando a cabo múltiples estudios con el fin de identificar agentes que sean efectivos ante COVID-19, en los siguientes objetivos estratégicos: inhibición de la entrada/fusión del virus (anticuerpos neutralizantes, inhibidores de proteasa de serina transmembrana 2, cloroquina, hidroxicloroquina y umifenovir); interrupción de la replicación viral (remdesivir, favipiravir, lopinavir/ritonavir e ivermectina) y supresión de la respuesta inflamatoria excesiva (corticosteroides, tocilizumab, e inmunoglobulina). Aún no existe un tratamiento efectivo y seguro contra COVID-19; los fármacos descritos en esta revisión se administran como uso compasivo de drogas, o bien, como parte de un ensayo clínico. La terapia de soporte continúa siendo el pilar del manejo de COVID-19.(AU)

There is no specific antiviral treatment for COVID-19. However, knowledge acquired during the SARS and MERS outbreaks, together with the information obtained with COVID-19, have allowed the detection of various therapeutic targets in the virus replication cycle, and in its pathogenesis. The current evidence regarding the leading treatments proposed for COVID-19, reused or experimental, is included through a review of the scientific literature to date. Due to the lack of randomized controlled trials, the following were involved: case reports, case series and review articles. Globally, multiple studies are being carried out in order to identify agents that are effective against COVID-19, upon the following strategic objectives: inhibition of viral entry/fusion (neutralizing antibodies, transmembrane serine protease 2 inhibitors, chloroquine, hydroxychloroquine, and umifenovir); interruption of viral replication (remdesivir, favipiravir, lopinavir/ritonavir and ivermectin), and suppression of excessive inflammatory response (corticosteroids, tocilizumab, and immunoglobulin). There is still no effective and safe treatment against COVID-19; the medications described in this review are given as compassionate drug use, or as part of a clinical trial. Support therapy continues to be COVID-19 management cornerstone.(AU)

Intervenciones farmacológicas para el tratamiento de la Enfermedad por Coronavirus (COVID-19) / Pharmacological interventions for the treatment of Coronavirus Disease (COVID-19)

RESUMEN Varios fármacos han sido propuestos como alternativas terapéuticas para COVID-19. Se efectuó una búsqueda sistematica en MEDLINE (vía PubMed) hasta el 20 de marzo de 2020, con el fin de identificar la evidencia disponible sobre intervenciones farmacológicas para tratamiento específico de COVID-19. 947 publicaciones fueron identificadas y 15 estudios seleccionados: 3 ensayos clínicos, 5 series de casos y 7 reportes de casos. La calidad de la evidencia procedente de ensayos clínicos fue evaluada según la metodología GRADE. La evidencia existente para hidroxicloroquina, favipiravir y lopinavir/ritonavir procede de ensayos clínicos que reportan resultados favorables para los dos primeros fármacos en tanto que no se observó ningún beneficio al adicionar lopinavir/ritonavir al tratamiento estándar. Sin embargo, debido a las limitaciones metodológicas, la evidencia es de muy baja certeza para hidroxicloroquina y de baja certeza para favipiravir y lopinavir/ritonavir. Respecto al uso de arbidol interferón, o el uso combinado de estos con lopinavir/ritonavir, la evidencia es limitada ya que deriva de serie de casos o reporte de casos con resultados no determinantes. No se identificaron estudios que permitan determinar la eficacia y seguridad de intervenciones farmacológicas frente a COVID-19.

ABSTRACT Several drugs have been proposed as therapeutic alternatives for COVID-19. An electronic systematic search of MEDLINE (via PubMed) was carried out until March 20th 2020, in order to identify the available evidence on pharmacological interventions for specific treatment of COVID-19. The quality of the evidence from clinical trials was evaluated according to the GRADE methodology. 947 publications were identified and 15 studies were selected: 3 clinical trials, 5 case series and 7 case reports. The existing evidence for hydroxychloroquine, favipiravir and lopinavir/ritonavir comes from clinical trials reporting favorable results for the first two drugs, while no benefit was observed when lopinavir/ritonavir was added to standard treatment. However, due to methodological issues, the evidence for hydroxychloroquine is very low. For favipiravir and lopinavir/ritonavir the evidence is low. Regarding the use of arbidol interferon, or the combined use of these with lopinavir/ritonavir, the evidence is limited since it derives from case series or case report with non-determining results. No studies were identified that reliably demonstrate the efficacy and safety of any pharmacological intervention against COVID-19.

COVID-19: En este momento podría ser útil el tratamiento antiviral en casos leves / COVID-19: Antiviral treatment in mild cases could be useful at this time

RESUMEN La letalidad por COVID-19 así como su rápida diseminación son responsables de la actual crisis mundial; por ello, desde un inicio se buscaron medicamentos con acción antiviral frente a este agente. Es imposible, por ahora, saber qué persona en estado leve desarrollará una elevada carga viral o tiene una predisposición a desarrollar una respuesta extremada del sistema inmune, pero un tratamiento temprano en los casos leves no sólo garantizaría una mayor eficacia, sino que se evitarían los casos severos. En la actualidad no se dispone de ensayos clínicos, doble ciego, aleatroizados, ni metaanálisis para tomar decisiones seguras; mientras tanto, la pandemia avanza en nuestro país, generando dolor y muerte. En este contexto, en el Perú, las sociedades científicas han manifestado la necesidad del uso de medicamentos antivirales, aún con poca evidencia, basado en la probabilidad de éxito previo a un tratamiento, por sus efectos in vitro y por sus efectos clínicos tempranos, tales como: cloroquina, hidroxicloroquina, azitromicina, lopinavir, ritonavir. De esta manera, en el país desde el 29 de marzo de 2020, se dispone de una norma técnica del Ministerio de Salud, que facilita el uso de estos a nivel nacional para los casos moderados y severos. Consideramos que bajo un sistema estructurado, los centros de primer nivel de atención podrían tratar los casos leves de COVID-19. Nuestro país, a través de las estrategias de prevención y control de TB y de ITS/VIH/SIDA, tiene experiencia en el manejo de programas de tratamiento en dicho nivel. Los costos serán menores que los requeridos en el fortalecimiento del tercer nivel de atención.

ABSTRACT COVID-19's lethality as well as its rapid spread are responsible for the current world crisis; therefore, from the beginning, drugs with antiviral action against this agent were sought. It is impossible, for now, to know which person in a mild state will develop a high viral load or have a predisposition to develop an extreme response from the immune system, but early treatment in mild cases would not only guarantee greater efficacy, but would avoid severe cases. Currently, there are no double blind, randomized clinical trials or meta-analyzes available to make safe decisions; meanwhile, the pandemic is advancing in our country, generating pain and death. In this context, in Peru, scientific societies have expressed the need for the use of antiviral drugs, even with little evidence, based on the probability of success prior to treatment, for their in vitro effects and for their early clinical effects, such as: chloroquine, hydroxychloroquine, azithromycin, lopinavir, ritonavir. Thus, in the country since march 29, 2020, there is a technical standard from the Ministry of Health, which facilitates the use of these at the national level for moderate and severe cases. We believe that under a structured system, first-level care centers could treat mild cases of COVID-19. Our country, through strategies for the prevention and control of TB and ITS/HIV/AIDS, has experience in managing treatment programs at this level. The costs will be less than those required to strengthen the third level of care.

Systematic review on current antiviral therapy in COVID-19 pandemic

@#Introduction: Currently, there are several attempts to find an effective antiviral drugs against the COVID-19. Although majority of the COVID-19 patients have mild to moderate clinical events, up to 5-10% may have severe, life threatening events that urgently require effective drugs. The purpose of this systematic review is to evaluate the effectiveness of antiviral therapies in the treatment of COVID-19. Methods: An extensive search was performed in PubMed, EMBASE, Cochrane Library for randomised controlled trials (RCTs), prospective case series studies that evaluated therapies COVID-19. The outcomes searched for were mortality, recovery rate, length of hospital stay and clinical improvement from January to May 15, 2020. Independent reviewers searched, identified, screened, and related studies were included. Results: Total of five RCTs on 439 patients and seventeen case series involving 1656 patients were found in the specified review period that reported the use of Lopinavir, Ritonavir, Remdesivir. Oseltamivir, Ribavirin in patients with COVID-19; but none of which showed efficacy of antiviral therapy. Such current findings impede researchers from recommending an appropriate and effective antiviral therapy against COVID-19, making it a serious concern for the global community. Discussion: In the present pandemic and any future epidemics, all the related authorities should pursue many more RCTs, cohort and case series for a prospective outcome in the management and treatment guidelines.

Clinical characteristics and treatment strategies of 79 patients with COVID-19 / 中国药理学通报

Aim To analyze the clinical characteristics of patients of corona virus disease 2019 (COVID-19), and summarize the treatment experience, aiming to provide diagnostic and treatment reference for the front-line clinicians. Methods Seventy-nine patients with COVID-19 admitted to the Infectious Hospital of Anhui Provincial Hospital from January 22 to February 18, 2020 were selected as the research subjects. There were 55 cases in general group and 24 cases in severe and critical group. The clinical data of the two groups were collected and compared, including general conditions, clinical symptoms, signs, laboratory tests, computed tomography imaging of the lungs and complications. Results The average age of 79 COVID-19 patients was 45. 1 ± 16. 6 years, and forty-five of them are males. The severe and critical group was older than the general group. Besides, there were more males and comorbidities. In terms of laboratory tests, the lymphocyte (LYM) count and albumin (A L B) decreased more significantly in the severe and critical groups. Moreover, the percentage of neutrophils (N E U), c-reactive protein (CRP), D-dimer, lactate dehydrogenase (L D H), troponin I (cTnl) and urea nitrogen (BUN) increased significantly. Among all the patients, the types of antiviral drugs in severe and critical group were significantly more than those in general group; moreover, the glucocorticoids, gamma globulin and oxygen inhalation by nasal catheter were used more frequently in severe and critical patients. By the time of February 18, one patient died of acute large area cerebral infarction, 34 patients were discharged from hospital, and the rest were still receiving treatment. Conclusions The elderly patients with multiple cardiovascular diseases are more likely to get severe and critical COVID-19. They usually combine multiple organ or system abnormalities. Clinicians should make timely judgment and adjust the treatment plan according to the clinical symptoms, signs and laboratory examination results.

Safety of protease inhibitors and Arbidol for SARS-CoV-2 pneumonia in Zhejiang Province, China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The aim of this study was to evaluate the safety of an antiviral regimen of protease inhibitors combined with Arbidol (umifenovir) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pneumonia patients. The genomic sequence of SARS-CoV-2 is highly homologous to that of SARS-CoV (Zhou et al., 2020). Previously published basic and clinical research on anti-SARS-CoV treatment found that lopinavir/ritonavir (LPV/r) could improve the prognosis of SARS patients (Chan et al., 2003; Chu et al., 2004). Darunavir (DRV) is another protease inhibitor that blocks the binding of SARS-CoV-2 to human angiotensin-converting enzyme 2 (Omotuyi et al., 2020). The broad-spectrum antiviral drug Arbidol (umifenovir) also shows in vitro anti-SARS-CoV activity (Khamitov et al., 2008).

Efficacies of lopinavir/ritonavir and abidol in the treatment of novel coronavirus pneumonia / 中华传染病杂志

Objective@#To evaluate the efficacies of lopinavir/ritonavir and abidol in the treatment of NCP.@*Methods@#The clinical data of 134 patients with NCP receiving treatment at Shanghai Public Health Clinical Center during Jan 20 to Feb 6, 2020 were retrospectively collected. All the patients received interferon-α2b spray and symptomatic supportive treatment, and 52 cases received oral lopinavir/ritonavir treatment, 34 cases received oral abidol treatment, the remaining 48 cases did not take any antiviral drugs. The efficacies at median day 7 among the three groups were compared by using Kruskal-Wallis or chi-square tests.@*Results@#The 134 patients included 69 males (51.5%) and 65 females, aged 35-62 years with the average of 48 years. The median time to temperature normalization in patients receiving abidol or lopinavir/ritonavir treatment was both 6 days after admission, and that was 4 days in the control group, with no significant difference (χ2=2.37, P=0.31). The median time to PCR negative in the respiratory specimens in the three groups was all 7 days after admission, and the PCR negative rates at day 7 after admission in lopinavir/ritonavir, abidol and control groups were 71.8% (28/39), 82.6% (19/23) and 77.1% (27/35), respectively, which were not significantly different (χ2=0.46, P=0.79). Radiological worsening at day 7 was observed in comparable proportions of patients in the three groups, which were 42.3% (n=22), 35.3% (n=12) and 52.1% (n=25), respectively (χ2=2.38, P=0.30) . Adverse reactions occurred in 17.3% (n=9), 8.8% (n=3) and 8.3% (n=4) patients, respectively in the three groups (χ2=2.33, P=0.33).@*Conclusions@#This study did not find any effects of lopinavir/ritonavir and abidol on relieving symptoms or accelerating virus clearance. The efficacies of these two drugs in NCP treatment need further investigation.

Early antiviral therapy of abidor combined with lopinavir/ritonavir and re-combinant interferonα-2b in patients with novel coronavirus pneumonia in Zhejiang: A multicenter and prospective study / 中华临床感染病杂志

Objective@#Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province.@*Methods@#A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data.@*Results@#The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05).@*Conclusions@#The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.

Lopinavir and curcumin directly alters BAX/BCL2 and VEFG165b mRNA levels to suppress human squamous cervical carcinoma cell growth / Lopinavir y curcumina alteran directamente los niveles de ARNm de BAX/ BCL 2 y VEFG165 para suprimir el crecimiento de células de carcinoma cervical escamoso humano

Following the success of the highly active antiretroviral therapy, the potential of multidrug combination regimen for the management of cancer is intensely researched. The anticancer effects of curcumin on some human cell lines have been documented. Lopinavir is a FDA approved protease inhibitor with known apoptotic activities. Dysregulated apoptosis is important for the initiation of cancer while angiogenesis is required for cancer growth and development, this study therefore investigated the effects of the combination of lopinavir and curcumin on cell viability, apoptosis and the mRNA expression levels of key apoptotic and angiogenic genes; BAX, BCL2 and VEGF165b in two human cervical cell lines; human squamous cell carcinoma cells - uterine cervix (HCS-2) and transformed normal human cervical cells (NCE16IIA). The two human cervical cell lines were treated with physiologically relevant concentrations of the agents for 120 h following which BAX, BCL2 and VEGF165b mRNA expression were determined by Real Time qPCR. The Acridine Orange staining for the morphological evaluation of apoptotic cells was also performed. The combination of lopinavir and curcumin up-regulated pro-apoptotic BAX and antiangiogenic VEGF165b but down-regulated the mRNA levels of anti-apoptotic BCL2 mRNA in the human squamous cell carcinoma (HCS-2) cells only. The fold changes were statistically significant. Micrographs from Acridine Orange staining showed characteristic evidence of apoptosis in the human squamous cell carcinoma (HCS-2) cells only. The findings reported here suggest that the combination of curcumin and the FDA approved drug-lopinavir modulate the apoptotic and angiogenic pathway towards the inhibition of cervical cancer.

Tras el éxito de la terapia antirretroviral altamente activa, se investiga intensamente el potencial del régimen de combinación de múltiples fármacos para el tratamiento del cáncer. Se han documentado los efectos anticancerígenos de la curcumina en algunas líneas celulares humanas. Lopinavir es un inhibidor de proteasa aprobado por la FDA con actividades apoptóticas conocidas. La apoptosis disrregulada es importante para el inicio del cáncer, mientras que la angiogénesis es necesaria para el crecimiento y desarrollo del cáncer. Por lo tanto, este estudio investigó los efectos de la combinación de lopinavir y curcumina sobre la viabilidad celular, la apoptosis y los niveles de expresión del ARNm de genes apoptóticos y angiogénicos clave: BAX, BCL2 y VEGF165b en dos líneas celulares cervicales humanas; células de carcinoma de células escamosas humanas: cérvix uterino (HCS-2) y células cervicales humanas transformadas (NCE16IIA). Las dos líneas celulares cervicales humanas se trataron con concentraciones fisiológicamente relevantes de los agentes durante 120 horas, después de lo cual la expresión de ARNm de BAX, BCL2 y VEGF165b se determinó mediante qPCR en tiempo real. También se realizó la tinción con naranja de acridina para la evaluación morfológica de células apoptóticas. La combinación de lopinavir y curcumina reguló incrementando BAX proapoptósicos y VEGF165b antiangiogénicos, pero reguló a la baja los niveles de ARNm del BCL2 antiapoptótico en células de carcinoma de células escamosas humanas (HCS-2) únicamente. Los cambios en el pliegue fueron estadísticamente significativos. Las micrografías de la tinción con naranja de acridina mostraron evidencia característica de apoptosis solo en las células del carcinoma de células escamosas humanas (HCS-2). Los hallazgos reportados aquí sugieren que la combinación de curcumina y el fármaco aprobado por la FDA lopinavir modulan la vía apoptótica y angiogénica hacia la inhibición del cáncer cervical.

Dual therapy with lopinavir/ritonavir plus lamivudine could be a viable alternative for antiretroviral-therapy-naive adults with HIV-1 infection regardless of HIV viral load or subgenotype in resource-limited settings: A randomised, open-label and non-inferiority study from China

Backgrounds: This randomised controlled, open-label, non-inferiority trial was conducted in antiretroviral-na飗e HIV-1-infected patients to assess the efficacy and safety of 48-week dual therapy of LPV/r plus 3TC (DT group) compared with Chinese first-line triple-therapy regimen (TT group). Methods: 198 were randomised to DT (n = 100) or TT (n = 98). Results: Ninety-two DT patients (92%) and 88 TT patients (89.8%) achieved HIV-1 RNA <50 copies/ml at week 48 (P = 0.629). Moreover, the safety profile was similar between two groups, and no secondary HIV resistance was observed. Conclusion: The results suggest that dual therapy of LPV/r plus 3TC is non-inferior to the first-line triple-therapy regimen in China.

Development of LC-MS/MS method for rapid quantification of lopinavir/ritonavir(Kaletra)in human plasma / 国际药学研究杂志

Objective To establish a rapid LC-MS/MS method for the simultaneous quantitative determination of lopinavir (LPV)and ritonavir(RTV)in human plasma. Methods Plasma samples were prepared by protein precipitation and separated by a Thermo Hypersil GOLD column(2.1 mm×100 mm,5 mm)with the mobile phase consisting of methanol and water(0.1%formic acid) at a flow rate of 0.2 ml/min. Detection of LPV,RTV and the internal standard(IS)MS 275 was performed using selected reaction moni?toring(SRM)of the transitions m/z 629.3→155.0,m/z 721.3→268.0 and m/z 377.1→359.2 in positive ion mode,respectively. Re?sults The calibration curve was linear in the range of 20-1000 ng/ml(r>0.994)for LPV and RTV. The intra and inter-day precision and accuracy values met the set acceptance criteria. Conclusion The method is rapid,sensitive and accurate for the therapeutic drug monitoring of LPV and RTV simultaneously in clinic and pharmacokinetic studies.

Development of LC-MS/MS method for rapid quantification of lopinavir/ritonavir (Kaletra) in human plasma / 国际药学研究杂志

Objective To establish a rapid LC- MS/MS method for the simultaneous quantitative determination of lopinavir (LPV) and ritonavir (RTV) in human plasma. Methods Plasma samples were prepared by protein precipitation and separated by a Thermo Hypersil GOLD column (2.1 mm×100 mm, 5 mm) with the mobile phase consisting of methanol and water (0.1% formic acid) at a flow rate of 0.2 ml/min. Detection of LPV, RTV and the internal standard (IS) MS 275 was performed using selected reaction moni-toring (SRM) of the transitions m/z 629.3→155.0, m/z 721.3→268.0 and m/z 377.1→359.2 in positive ion mode, respectively. Re-sults The calibration curve was linear in the range of 20-1000 ng/ml (r0.994) for LPV and RTV. The intra and inter-day precision and accuracy values met the set acceptance criteria. Conclusion The method is rapid, sensitive and accurate for the therapeutic drug monitoring of LPV and RTV simultaneously in clinic and pharmacokinetic studies.

Pharmacokinetic profiles of lopinavir(LPV) in Chinese HIV-infected patients / 中华内科杂志

Objective To evaluate the pharmacokinetic profiles of lopinavir(LPV) in Chinese HIV-infected patients.Methods A total of 16 patients were enrolled in the LPV pharmacokinetic study.Blood samples were collected before LPV intake and 0.5,1.0,1.5,2.0,2.5,3.0,4.0,6.0,8.0,10.0,12.0 h after administration.Serum level of LPV was determined by the developed high performance liquid chromatography (HPLC) method.The pharmacokinetic profiles were assessed by WinNonlin software.Results The non-compartment model pharmacokinetic (PK) parameters were as follows:the peak time of LPV (Tmax) (3.88 ± 0.23) h,maximum plasma concentration (Cmax) (10.36 ± 3.42) mg/L,minimum plasma concentration (Cmin) (2.18 ± 0.34) mg/L,the 24 h area under plasma-concentration-time curve (AUC0-24) (116.22 ± 15.68) mg · h · L-1,half life(T1/2) (4.5 ± 0.13) h,and clearance rate (CL/F) (3.44 ± 1.34) L/h respectively.Conclusions The pharmacokinetic profiles of LPV in Chinese HIV-1 infected patients demonstrate lower Cmin than those of reported studies,while other parameters are similar.Patients should be educated for compliance based on the narrow gap between Cmin and minimum effect concentration.