A simple, precise and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the simultaneous determination of oseltamivir and oseltamivir carboxylate, a neuramin...A simple, precise and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the simultaneous determination of oseltamivir and oseltamivir carboxylate, a neuraminidase inhibitor, using their deuterated analogs as internal standards （ISs）. The method involved solid phase extraction of the analytes and ISs from 200 μL human plasma with no reconstitution and drying steps. The chromatographic separation was achieved on a Symmetry C18 （100 mm × 4.6 mm, 5 μm） column using 10 mM ammonium formate and acetonitrile （30：70, v/v） as the mobile phase in a run time of 2.0 min. Quantitation of analytes and ISs were done by multiple reaction monitoring on a triple quadrupole mass spectrometer in the positive ionization mode. The linearity of the method was established in the concentration range of 0.5-200 ng/mL and 2.0-800 ng/mL for oseltamivir and oseltamivir carboxylate respectively. The mean extraction recovery for oseltamivir （94.4%） and oseltamivir carboxylate （92.7%） from spiked plasma samples was consistent and reproducible. The application of this method was demonstratedby a bioequivalence study in 42 healthy Indian subjects with 75 mg oseltamivir phosphate capsules. The assay reproducibility was established by reanalysis of 151 incurred subject samples.展开更多
The use of oseltamivir, widely stockpiled as one of the drugs for use in a possible avian influenza pandemic, has been reported to be associated with neuropsychiatric disorders and severe skin reactions, primarily in ...The use of oseltamivir, widely stockpiled as one of the drugs for use in a possible avian influenza pandemic, has been reported to be associated with neuropsychiatric disorders and severe skin reactions, primarily in Japan. Here we identified a nonsynonymous SNP （single nucleotide polymorphism） in dbSNP database, R41Q, near the enzymatic active site of human cytosolic sialidase, a homologue of virus neuraminidase that is the target of oseltamivir. This SNP occurred in 9.29% of Asian population and none of European and African American population. Our structural analyses and Ki measurements using in vitro sialidase assays indicated that this SNP could increase the unintended binding affinity of human sialidase to oseltamivir carboxylate, the active form of oseltamivir, thus reducing sialidase activity. In addition, this SNP itself results in an enzyme with an intrinsically lower sialidase activity, as shown by its increased Km and decreased Vmax values. Theoretically administration of oseltamivir to people with this SNP might further reduce their sialidase activity. We note the similarity between the reported neuropsychiatric side effects ofoseltamivir and the known symptoms of human sialidase-related disorders. We propose that this Asian-enriched sialidase variation caused by the SNP, likely in homozygous form, may be associated with certain severe adverse reactions to oseltamivir.展开更多
Objective To evaluate the bioavailability and bioequivalence of oseltamivir capsule in Chinese health male volunteers.Methods A randomized,two period,two treatment,two sequence crossover bioequivalence trial was desig...Objective To evaluate the bioavailability and bioequivalence of oseltamivir capsule in Chinese health male volunteers.Methods A randomized,two period,two treatment,two sequence crossover bioequivalence trial was designed,24 Chinese health volunteers were randomly divided into two groups,each group was orally given single dose oseltamivir phosphate(tamifla)or AMMS 607 capsule.The active metabolite oseltamivir carboxylate of oseltamivir in the plasma were determined by liquid chromatographic-tandem mass spectrometric(LC-MS/MS)method.The pharmacokinetics parameters and relative bioavailability were calculated to evaluate the bioequivalence of AMMS 607 and tamifla.Results Cmax of the AMMS 607 and tamifla were 602.07±153.27 ng·mL-1 and 620.09±132.39 ng·mL-1 respectively;tmax were 4.2±1.1 h and 4.8±1.0 h;t1/2β were 6.60±0.87 h and 6.61±0.83 h;MRT were 10.00±1.77 h and 10.40±1.62 h;AUC0-24 were 6285.88±1083.66 ng·h·mL-1 and 6546.01±1199.32 ng·h·mL-1;Compared with the reference of tamifla capsule,the bioavailability F0-tn of AMMS 607 capsule was 99.5±27.7%.The main pharmacokinetics parameters of AUC0-24,Cmax and Tmax showed no statistically significant difference between the two capsules.Conclusions The AMMS 607 capsule and tamifla capsule are bioequivalent.展开更多
This research focused on the modification of the functional groups of oseltamivir as neuraminidase inhibitor against influenza A virus subtype H1N1.Interactions of three of the best ligands were evaluated in the hydra...This research focused on the modification of the functional groups of oseltamivir as neuraminidase inhibitor against influenza A virus subtype H1N1.Interactions of three of the best ligands were evaluated in the hydrated state using molecular dynamics simulation at two different temperatures.The docking result showed that AD3BF2 D ligand（N-[（1S,6R）-5-amino-5-{[（2R,3S,4S）-3,4-dihydroxy-4-（hydroxymethyl） tetrahydrofuran-2-yl]oxy}-4-formylcyclohex-3-en-1-yl]acetamide-3-（1-ethylpropoxy）-1-cyclohexene-1-carboxylate） had better binding energy values than standard oseltamivir.AD3BF2 D had several interactions,including hydrogen bonds,with the residues in the catalytic site of neuraminidase as identified by molecular dynamics simulation.The results showed that AD3BF2 D ligand can be used as a good candidate for neuraminidase inhibitor to cope with influenza A virus subtype H1N1.展开更多
Influenza A virus poses a great threat to global health, and oseltamivir (trade marked as Tamiflu), which targets influenza surface glycoprotein neuraminidase (NA), is used clinically as a major anti-influenza treatme...Influenza A virus poses a great threat to global health, and oseltamivir (trade marked as Tamiflu), which targets influenza surface glycoprotein neuraminidase (NA), is used clinically as a major anti-influenza treatment. However, certain substitutions in NA can render an influenza virus resistant to this drug. In this study, using a lentiviral pseudotyping system, which alleviates the safety concerns of studying highly pathogenic influenza viruses such as avian influenza H5N1, that utilizes influenza surface glycoproteins (hemagglutinin or HA, and NA) and an HIV-core combined with a luciferase reporter gene as a surrogate assay, we first assessed the functionality of NA by measuring pseudovirion release in the absence or presence of oseltamivir. We demonstrated that oseltamivir displays a dose-dependent inhibition on NA activity. In contrast, a mutant NA (H274Y) is more resistant to oseltamivir treatment. In addition, the effects of several previously reported substitution NA mutants were examined as well. Our results demonstrate that this lentivirus-based pseudotyping system provides a quick, safe, and effective way to assess resistance to neuraminidase inhibitors. And we believe that as new mutations appear in influenza isolates, their impact on the effectiveness of current and future anti-NA can be quickly and reliably evaluated by this assay.展开更多
Objective This study aimed to assess the efficacy and safety of traditional Chinese medicine,alone or in combination with oseltamivir,in patients with H1N1 Influenza.Methods In the present study,we searched the Cochra...Objective This study aimed to assess the efficacy and safety of traditional Chinese medicine,alone or in combination with oseltamivir,in patients with H1N1 Influenza.Methods In the present study,we searched the Cochrane Central Register of Controlled Trials,PUBMED,EMBASE,Chinese Biomedical Literature Database,China Science and Technology Journal Database,China National Knowledge Infrastructure Database,and WanFang Data for studies published in or before February 8,2022.Data were extracted and checked by two investigators.Review Manager 5.4 and STATA statistical software 16.0 were used for the data analysis.Results We identified 22 individual studies reporting data from 2292 individuals with H1N1 influenza.Compared with oseltamivir,the fever clearance duration[MD=-3.99,95%CI(-6.89,-1.09)]and sore throat relief time[MD=-5.39,95%CI(-10.19,-0.59)]in the intervention group of traditional Chinese medicine monotherapy or combined with oseltamivir were shorter.Maxingshigan was the primary component of Lianhuaqingwen.The subgroup analyses indicated that Maxingshigan shortened fever clearance time[MD=-3.23,95%CI(-5.60,-0.85)],and also had certain advantages in relieving sore throat[RR=-4.55,95%CI(-10.04,0.95)].However,as for the effective rate,fever duration,cough disappearance time,hospital length of stay,clinical symptoms time as well as viral shedding duration,there were no significant differences between the two groups.Besides,no serious adverse effects were reported in the included studies.Conclusion Although we couldn’t get a definitive conclusion due to the small sample sizes and high risk of bias in the included studies,most traditional Chinese medicine showed similar effects to oseltamivir in treating H1N1 influenza.Some were showed to have a statistically significant shorter time of fever clearance and sore throat remission when they were used alone or in combination with oseltamivir and were well-tolerated treatment,such as Maxingshigan.展开更多
Anti-viral chemotherapy plays an important part in treating and preventing influenza illness. However, its effectiveness in severe infections can be debated and a reoccurring problem is the emergence of resistant viru...Anti-viral chemotherapy plays an important part in treating and preventing influenza illness. However, its effectiveness in severe infections can be debated and a reoccurring problem is the emergence of resistant virus. Passive immunisation has for a long time been and is still used for prophylaxis and treatment of a number of infectious diseases. In this experimental study anti-influenza antibodies were passively administrated to mice, subsequently they were infected with influenza virus and treated with oseltamivir. The aim was to investigate, if anti-influenza antibodies influenced the out come of oseltamivir treatment and development of resistance towards oseltamivir. We show, that oseltamivir alone was not able to effectively prevent a fatal outcome, but that oseltamivir administered together with a limited amount of antibodies, resulted in improvement of the clinical condition of the mice. The results also showed that a higher dosage of antibodies alone were able to protect the mice from a lethal dose of virus. These findings suggest that the effectiveness of oseltamivir depends on the host’s immune response to the influenza virus, and that that passive immunization is an option that should be considered in the in control of influenza.展开更多
Background:The resurgence of seasonal influenza virus circulation has been seen in 2021-2022 after the tempo-rary suppression in 2020-2021.Neuraminidase inhibitors(NAIs)are widely applied in the clinical treatment of ...Background:The resurgence of seasonal influenza virus circulation has been seen in 2021-2022 after the tempo-rary suppression in 2020-2021.Neuraminidase inhibitors(NAIs)are widely applied in the clinical treatment of influenza A despite several limitations.Objective:To access the efficacy of Xu’s influenza decoction(XID)in combination therapy with oseltamivir for the treatment of influenza A.Methods:In this retrospective cohort study,the eligible participants were diagnosed with influenza A between June 1,2018,and May 30,2022,in the First Affiliated Hospital of Zhejiang Chinese Medical University.According to whether Xu’s influenza decoction was applied,patients were divided into two groups:treated with or without XID.Propensity score matching(PSM)was used to further adjust the covariates between groups.The primary outcome was to compare time to defervescence via K-M curves,Breslow tests,and Cox regression analysis.In Cox proportional hazards model,a univariate analysis was performed to obtain preliminary results,while a further multivariate analysis was conducted to study the independent factors that influence defervescence.Subgroup analysis was conducted according to body temperature and time from onset to admission.The secondary outcome consisted of routine blood and C-reactive protein(CRP),length of stay,and medical costs.Results:A total of 336 patients with influenza A were enrolled in this study(i.e.,163 patients in the XID+oseltamivir group;173 patients in the oseltamivir group).After 1:1 matching via PSM,230 patients meeting the criteria were included in the analysis,with 115 in each arm.The XID+oseltamivir group had shorter time to defervescence(36 h vs 44 h,P=0.011),shorter length of stay(3 days vs 4 days,P=0.018),and higher defervescence possibility(HR=1.384,95%CI:1.054-1.818).Subgroup analysis indicated that for patients during non-window period(≥48 h)with medium-grade fever(38.1℃-39℃),the XID+oseltamivir combination therapy reduced time to defervescence(P=0.04995/0.004)with a higher defervescence possibility(HR=1.524/1.683).Meanwhile,there’s no statistical significance but observable trends of the XID+oseltamivir group in the lower medical costs(3068.07 yuan vs 3120.68 yuan),the lower neutrophils%(48.53%vs 51.00%)and the higher lymphocyte%(39.83%vs 37.72%).Conclusion:The combination of XID and oseltamivir can shorten the time to defervescence and length of stay in influenza A.Its antipyretic effect is mainly reflected in the medium-grade and non-window periods.展开更多
文摘A simple, precise and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the simultaneous determination of oseltamivir and oseltamivir carboxylate, a neuraminidase inhibitor, using their deuterated analogs as internal standards （ISs）. The method involved solid phase extraction of the analytes and ISs from 200 μL human plasma with no reconstitution and drying steps. The chromatographic separation was achieved on a Symmetry C18 （100 mm × 4.6 mm, 5 μm） column using 10 mM ammonium formate and acetonitrile （30：70, v/v） as the mobile phase in a run time of 2.0 min. Quantitation of analytes and ISs were done by multiple reaction monitoring on a triple quadrupole mass spectrometer in the positive ionization mode. The linearity of the method was established in the concentration range of 0.5-200 ng/mL and 2.0-800 ng/mL for oseltamivir and oseltamivir carboxylate respectively. The mean extraction recovery for oseltamivir （94.4%） and oseltamivir carboxylate （92.7%） from spiked plasma samples was consistent and reproducible. The application of this method was demonstratedby a bioequivalence study in 42 healthy Indian subjects with 75 mg oseltamivir phosphate capsules. The assay reproducibility was established by reanalysis of 151 incurred subject samples.
文摘The use of oseltamivir, widely stockpiled as one of the drugs for use in a possible avian influenza pandemic, has been reported to be associated with neuropsychiatric disorders and severe skin reactions, primarily in Japan. Here we identified a nonsynonymous SNP （single nucleotide polymorphism） in dbSNP database, R41Q, near the enzymatic active site of human cytosolic sialidase, a homologue of virus neuraminidase that is the target of oseltamivir. This SNP occurred in 9.29% of Asian population and none of European and African American population. Our structural analyses and Ki measurements using in vitro sialidase assays indicated that this SNP could increase the unintended binding affinity of human sialidase to oseltamivir carboxylate, the active form of oseltamivir, thus reducing sialidase activity. In addition, this SNP itself results in an enzyme with an intrinsically lower sialidase activity, as shown by its increased Km and decreased Vmax values. Theoretically administration of oseltamivir to people with this SNP might further reduce their sialidase activity. We note the similarity between the reported neuropsychiatric side effects ofoseltamivir and the known symptoms of human sialidase-related disorders. We propose that this Asian-enriched sialidase variation caused by the SNP, likely in homozygous form, may be associated with certain severe adverse reactions to oseltamivir.
文摘Objective To evaluate the bioavailability and bioequivalence of oseltamivir capsule in Chinese health male volunteers.Methods A randomized,two period,two treatment,two sequence crossover bioequivalence trial was designed,24 Chinese health volunteers were randomly divided into two groups,each group was orally given single dose oseltamivir phosphate(tamifla)or AMMS 607 capsule.The active metabolite oseltamivir carboxylate of oseltamivir in the plasma were determined by liquid chromatographic-tandem mass spectrometric(LC-MS/MS)method.The pharmacokinetics parameters and relative bioavailability were calculated to evaluate the bioequivalence of AMMS 607 and tamifla.Results Cmax of the AMMS 607 and tamifla were 602.07±153.27 ng·mL-1 and 620.09±132.39 ng·mL-1 respectively;tmax were 4.2±1.1 h and 4.8±1.0 h;t1/2β were 6.60±0.87 h and 6.61±0.83 h;MRT were 10.00±1.77 h and 10.40±1.62 h;AUC0-24 were 6285.88±1083.66 ng·h·mL-1 and 6546.01±1199.32 ng·h·mL-1;Compared with the reference of tamifla capsule,the bioavailability F0-tn of AMMS 607 capsule was 99.5±27.7%.The main pharmacokinetics parameters of AUC0-24,Cmax and Tmax showed no statistically significant difference between the two capsules.Conclusions The AMMS 607 capsule and tamifla capsule are bioequivalent.
文摘This research focused on the modification of the functional groups of oseltamivir as neuraminidase inhibitor against influenza A virus subtype H1N1.Interactions of three of the best ligands were evaluated in the hydrated state using molecular dynamics simulation at two different temperatures.The docking result showed that AD3BF2 D ligand（N-[（1S,6R）-5-amino-5-{[（2R,3S,4S）-3,4-dihydroxy-4-（hydroxymethyl） tetrahydrofuran-2-yl]oxy}-4-formylcyclohex-3-en-1-yl]acetamide-3-（1-ethylpropoxy）-1-cyclohexene-1-carboxylate） had better binding energy values than standard oseltamivir.AD3BF2 D had several interactions,including hydrogen bonds,with the residues in the catalytic site of neuraminidase as identified by molecular dynamics simulation.The results showed that AD3BF2 D ligand can be used as a good candidate for neuraminidase inhibitor to cope with influenza A virus subtype H1N1.
文摘Influenza A virus poses a great threat to global health, and oseltamivir (trade marked as Tamiflu), which targets influenza surface glycoprotein neuraminidase (NA), is used clinically as a major anti-influenza treatment. However, certain substitutions in NA can render an influenza virus resistant to this drug. In this study, using a lentiviral pseudotyping system, which alleviates the safety concerns of studying highly pathogenic influenza viruses such as avian influenza H5N1, that utilizes influenza surface glycoproteins (hemagglutinin or HA, and NA) and an HIV-core combined with a luciferase reporter gene as a surrogate assay, we first assessed the functionality of NA by measuring pseudovirion release in the absence or presence of oseltamivir. We demonstrated that oseltamivir displays a dose-dependent inhibition on NA activity. In contrast, a mutant NA (H274Y) is more resistant to oseltamivir treatment. In addition, the effects of several previously reported substitution NA mutants were examined as well. Our results demonstrate that this lentivirus-based pseudotyping system provides a quick, safe, and effective way to assess resistance to neuraminidase inhibitors. And we believe that as new mutations appear in influenza isolates, their impact on the effectiveness of current and future anti-NA can be quickly and reliably evaluated by this assay.
基金This work was supported by the National Administration of Traditional Chinese Medicine Project(2019XZZX-LG04)Chinese Academy of Traditional Chinese Medicine Project(ZZ13-035-02)to S.L.
文摘Objective This study aimed to assess the efficacy and safety of traditional Chinese medicine,alone or in combination with oseltamivir,in patients with H1N1 Influenza.Methods In the present study,we searched the Cochrane Central Register of Controlled Trials,PUBMED,EMBASE,Chinese Biomedical Literature Database,China Science and Technology Journal Database,China National Knowledge Infrastructure Database,and WanFang Data for studies published in or before February 8,2022.Data were extracted and checked by two investigators.Review Manager 5.4 and STATA statistical software 16.0 were used for the data analysis.Results We identified 22 individual studies reporting data from 2292 individuals with H1N1 influenza.Compared with oseltamivir,the fever clearance duration[MD=-3.99,95%CI(-6.89,-1.09)]and sore throat relief time[MD=-5.39,95%CI(-10.19,-0.59)]in the intervention group of traditional Chinese medicine monotherapy or combined with oseltamivir were shorter.Maxingshigan was the primary component of Lianhuaqingwen.The subgroup analyses indicated that Maxingshigan shortened fever clearance time[MD=-3.23,95%CI(-5.60,-0.85)],and also had certain advantages in relieving sore throat[RR=-4.55,95%CI(-10.04,0.95)].However,as for the effective rate,fever duration,cough disappearance time,hospital length of stay,clinical symptoms time as well as viral shedding duration,there were no significant differences between the two groups.Besides,no serious adverse effects were reported in the included studies.Conclusion Although we couldn’t get a definitive conclusion due to the small sample sizes and high risk of bias in the included studies,most traditional Chinese medicine showed similar effects to oseltamivir in treating H1N1 influenza.Some were showed to have a statistically significant shorter time of fever clearance and sore throat remission when they were used alone or in combination with oseltamivir and were well-tolerated treatment,such as Maxingshigan.
文摘Anti-viral chemotherapy plays an important part in treating and preventing influenza illness. However, its effectiveness in severe infections can be debated and a reoccurring problem is the emergence of resistant virus. Passive immunisation has for a long time been and is still used for prophylaxis and treatment of a number of infectious diseases. In this experimental study anti-influenza antibodies were passively administrated to mice, subsequently they were infected with influenza virus and treated with oseltamivir. The aim was to investigate, if anti-influenza antibodies influenced the out come of oseltamivir treatment and development of resistance towards oseltamivir. We show, that oseltamivir alone was not able to effectively prevent a fatal outcome, but that oseltamivir administered together with a limited amount of antibodies, resulted in improvement of the clinical condition of the mice. The results also showed that a higher dosage of antibodies alone were able to protect the mice from a lethal dose of virus. These findings suggest that the effectiveness of oseltamivir depends on the host’s immune response to the influenza virus, and that that passive immunization is an option that should be considered in the in control of influenza.
基金This work was financially supported by the General Project of Zhe-jiang Provincial Department of Education[Number:Y202248699]the Science and Technology Project of Zhejiang Provincial Administra-tion of Traditional Chinese Medicine[Number:2023ZL049].
文摘Background:The resurgence of seasonal influenza virus circulation has been seen in 2021-2022 after the tempo-rary suppression in 2020-2021.Neuraminidase inhibitors(NAIs)are widely applied in the clinical treatment of influenza A despite several limitations.Objective:To access the efficacy of Xu’s influenza decoction(XID)in combination therapy with oseltamivir for the treatment of influenza A.Methods:In this retrospective cohort study,the eligible participants were diagnosed with influenza A between June 1,2018,and May 30,2022,in the First Affiliated Hospital of Zhejiang Chinese Medical University.According to whether Xu’s influenza decoction was applied,patients were divided into two groups:treated with or without XID.Propensity score matching(PSM)was used to further adjust the covariates between groups.The primary outcome was to compare time to defervescence via K-M curves,Breslow tests,and Cox regression analysis.In Cox proportional hazards model,a univariate analysis was performed to obtain preliminary results,while a further multivariate analysis was conducted to study the independent factors that influence defervescence.Subgroup analysis was conducted according to body temperature and time from onset to admission.The secondary outcome consisted of routine blood and C-reactive protein(CRP),length of stay,and medical costs.Results:A total of 336 patients with influenza A were enrolled in this study(i.e.,163 patients in the XID+oseltamivir group;173 patients in the oseltamivir group).After 1:1 matching via PSM,230 patients meeting the criteria were included in the analysis,with 115 in each arm.The XID+oseltamivir group had shorter time to defervescence(36 h vs 44 h,P=0.011),shorter length of stay(3 days vs 4 days,P=0.018),and higher defervescence possibility(HR=1.384,95%CI:1.054-1.818).Subgroup analysis indicated that for patients during non-window period(≥48 h)with medium-grade fever(38.1℃-39℃),the XID+oseltamivir combination therapy reduced time to defervescence(P=0.04995/0.004)with a higher defervescence possibility(HR=1.524/1.683).Meanwhile,there’s no statistical significance but observable trends of the XID+oseltamivir group in the lower medical costs(3068.07 yuan vs 3120.68 yuan),the lower neutrophils%(48.53%vs 51.00%)and the higher lymphocyte%(39.83%vs 37.72%).Conclusion:The combination of XID and oseltamivir can shorten the time to defervescence and length of stay in influenza A.Its antipyretic effect is mainly reflected in the medium-grade and non-window periods.
