抗流感病毒新药oseltamivir的合成改进

目的 :优化抗流感病毒新药oseltamivir的手性全合成路线。方法 :以 (- )奎尼酸为起始物 ,经 12步立体选择性反应形成产物。结果 :得到的样品经FT IR ,1H NMR和MS波谱分析 ,证明是oseltamivir磷酸盐。结论 :本方法反应条件温和 ,提高了产率且降低了成本。

Simultaneous quantification of prodrug oseltamivir and its metabolite oseltamivir carboxylate in human plasma by LC-MS/MS to support a bioequivalence study

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.

抗流感病毒药物——Oseltamivir研究进展

流感是由流感病毒引起的一种急性呼吸道传染病,严重危害人类的健康和生命.根据流感病毒核蛋白(NP)和膜蛋白(MP)的不同,可将流感病毒分为A、B、C三型,A型常以流行的形式出现,B型常造成局部暴发,C型则多散发.

A nonsynonymous SNP in human cytosolic sialidase in a small Asian population results in reduced enzyme activity: potential link with severe adverse reactions to oseltamivir

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.

The study of bioavailability and bioequivalence of oseltamivir in Chinese health volunteers

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.

In silico modification of oseltamivir as neuraminidase inhibitor of influenza A virus subtype H1N1

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.

Analysis of Oseltamivir Resistance Substitutions in Influenza Virus Glycoprotein Neuraminidase using a Lentivirus-Based Surrogate Assay System

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.

Traditional Chinese medicine as monotherapy or combined with oseltamivir in the treatment of H1N1 influenza:a systematic review and meta-analysis

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.

口服oseltamivir治疗小儿流行性感冒

背景：口服oseltamivir是成人流行性感冒的有效治疗方案，本研究是为了确定该药治疗小儿流行性感冒的有效性、安全性和耐受程度。方法：在此项随机、双盲、安慰剂对照研究中，患有发热[≥100°F(38℃）]、咳嗽、鼻卡他症状，发热48小时以内的1～12岁儿童接受oseltamivir 2mg/kg/次，每日两次，或安慰剂，疗程5日。主要的判断有效终点是：解除疾病的时间即无或只有轻度咳嗽，无或只有轻度鼻卡他，并恢复了正常活动。结果：695例登记者中有452例（65%）证实为流行性感冒（安慰剂235例，oseltamivir217例）。在受感染患儿中接受oseltamivir治疗者与对照组相比其病程平均期限减少36小时（265）（101小时；95%可信区间，89～118vs.137小时；95%可信区间，125～150;P<0.0001=oseltamivir治疗也减轻了咳嗽、鼻炎和发热时期，中耳炎的新诊断减少44%（12%vs21%），治疗组的抗生素处方（68/217;30%）明显少于对照组（97/235，40%；P=0.03）。患儿接受oseltamivir治疗后哎吐发生虽较多（5.8%），但一般对此药耐受性较好。因副作用而停用治疗者在两组均很少（oseltamivir1.8%，安慰剂1.1%）。Oseltamivir对流感特异性抗体反应无影响。结论：流感患儿在发病48小时之内口服oseltamivir是一个有效且有良好耐受性的治疗方案。

生产oseltamivir为禽流感大流行作准备

在一次新闻发布会上，Roche公司宣布将与政府及制药公司代表协商，商讨生产oseltamivir（Ⅰ）以应付禽流感紧急流行时的需求。

Antibody administration in experimental influenza increases survival and enhances the effect of oseltamivir

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.

Oseltamivir可降低甲乙型流感并发症

流行性感冒对人类健康和整个社会而言都是一种严重的威胁；此外，流感还给病人造成严重的后果。据报道，在1997年美国的主要死因中，流感居第6位；仅美国每年约有36155例患者的死亡与流感相关的并发症有关。处于流感并发症高危的病人群体包括65岁以上的老年人与具有心血管或肺疾病史、免疫抑制或其他慢性疾病的患者。

The Effect of Xu’s Influenza Decoction Combined with Oseltamivir on Influenza A:A Propensity Score Matching Study

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.

感冒清热片防治甲型H1N1流感病毒感染的药效学研究

目的探讨感冒清热片防治甲型H1N1流感病毒感染的药效学情况。方法108例甲型H1N1流感病毒感染患者,随机分为观察组和对照组,每组54例。对照组使用磷酸奥司他韦治疗,观察组在对照组治疗基础上使用感冒清热片治疗。比较两组患者临床疗效、症状改善指标(起效时间、退热时间、止咳时间、痊愈时间)、以及治疗前后的中医证候(发热、咽痛、头痛、鼻塞流涕、咳嗽、乏力)积分、血清炎性因子[C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)]。结果观察组治疗总有效率96.30%高于对照组的77.78%(P<0.05)。治疗后,观察组患者发热、咽痛、头痛、鼻塞流涕、咳嗽、乏力评分分别为(0.83±0.24)、(1.12±0.30)、(1.07±0.29)、(1.03±0.32)、(1.05±0.33)、(1.09±0.28)分,均低于对照组的(1.46±0.37)、(2.31±0.58)、(2.14±0.52)、(2.25±0.63)、(2.36±0.61)、(2.42±0.65)分(P<0.05)。观察组起效时间(1.03±0.34)d、退热时间(1.56±0.40)d、止咳时间(4.35±0.68)d、痊愈时间(6.63±1.26)d均短于对照组的(2.32±0.57)、(2.63±0.74)、(7.24±1.16)、(9.38±1.72)d(P<0.05)。治疗后,观察组CRP(2.11±0.56)mg/L、TNF-α(19.34±2.20)μg/L、IL-6(4.02±0.54)ng/L均低于对照组的(4.83±0.69)mg/L、(25.89±2.73)μg/L、(6.13±0.62)ng/L(P<0.05)。结论感冒清热片防治甲型H1N1流感病毒感染的疗效显著,能有效缓解证候,加快症状缓解速度,提高抗炎作用。

磷酸奥司他韦联合丙种球蛋白治疗儿童ITP及对血小板相关指标、体液免疫、血栓相关指标的影响

目的研究磷酸奥司他韦联合丙种球蛋白治疗儿童免疫性血小板减少症(ITP)及对血小板相关指标、体液免疫、血栓相关指标的影响。方法前瞻性选择2022年6月至2023年6月于沧州市中心医院儿科治疗的ITP患儿60例作为研究对象。按照治疗方法不同将所有ITP患儿分为实验组(n=30)、对照组(n=30)。对照组给予丙种球蛋白治疗,实验组给予磷酸奥司他韦联合丙种球蛋白治疗,两组均治疗14 d。观察两组治疗前及治疗14 d后血小板相关指标[血小板计数(PLT)、血小板平均体积(MPV)、血小板膜糖蛋白CD42b(CD42b)、血小板膜糖蛋白PAC-1(PAC-1)]、血清体液免疫指标(IgA、IgG及IgM)、血清血栓相关指标[纤溶酶原激活抑制物-1(PAI-1)、凝血酶调节蛋白(TM)]和不良反应。采用Logistic回归分析影响磷酸奥司他韦联合丙种球蛋白治疗儿童ITP疗效的因素。结果实验组总有效率为90.00%,高于对照组(60.00%),差异有统计学意义(P<0.05)。治疗14 d后,实验组PLT水平及CD42b、PAC-1表达水平分别为(316.96±34.38)×10^(9)、94.09±9.72、5.59±0.58,均高于对照组[(252.07±28.16)×10^(9)、83.08±8.61、4.49±0.47],MPV水平为(9.95±0.98)fe,低于对照组[(10.94±1.27)fe],差异均有统计学意义(P<0.05)。治疗14 d后,实验组血清IgG水平为(9.18±0.94)g/L,低于对照组[(11.57±1.36)g/L],差异有统计学意义(P<0.05);两组血清IgA及IgM水平比较,差异均无统计学意义(P>0.05)。治疗14 d后,实验组血清PAI-1、TM水平分别为(32.97±3.46)μg/L、(88.06±9.13)ng/mL,均低于对照组[(51.98±5.32)μg/L、(102.95±12.27)ng/mL],差异均有统计学意义(P<0.05)。两组不良反应发生率比较,差异无统计学意义(P>0.05)。不同疗效ITP患儿的性别、出血情况、血小板计数、诱因间比较,差异均无统计学意义(P>0.05);年龄、病程间差异有统计学意义(P<0.05)。经Logistic多因素回归分析,年龄、病程不是影响磷酸奥司他韦联合丙种球蛋白治疗儿童ITP疗效的因素(P<0.05)。结论磷酸奥司他韦联合丙种球蛋白治疗ITP患儿可有效保护PLT,纠正异常体液免疫,改善血栓相关指标水平,具有较高的安全性,且其疗效不受性别、年龄、病程、出血情况、血小板计数、诱因等因素的影响。

基于FAERS的儿童奥司他韦不良事件信号挖掘与分析

目的 为临床安全使用奥司他韦提供参考。方法 检索美国食品和药物管理局不良事件报告系统(FAERS)数据库中2004年第1季度至2023年第1季度以奥司他韦为首要怀疑药物、发生于0~17岁患儿的药品不良事件(ADE)报告,采用报告比值比(ROR)法、比例报告比值比(PRR)法进行信号挖掘,采用《国际医学用语词典》中的首选语(PT)及系统器官分类(SOC)进行编码和归类。结果 共挖掘到奥司他韦相关ADE报告3 100份,检测到阳性信号76个,主要集中在精神病类(34个)、各类神经系统疾病(13个)、胃肠系统疾病(7个)、全身性疾病及给药部位各种反应(6个)、免疫系统疾病(3个)等12个SOC。发生频次排前20位的ADE有幻觉(455次)、异常行为(428次)、呕吐(410次)、谵妄(120次)、意识模糊状态(111次)等,信号强度排前20位的ADE仍以精神病类、各类神经系统疾病、胃肠系统疾病为主。发生频次和信号强度均排前20位的ADE有尖叫、恐惧、睡惊症,药品说明书中均未提及。结论 患儿临床应用奥司他韦时,应关注精神疾病、神经系统和胃肠道系统,以及皮肤和眼部的ADE。

泉州清新方联合奥司他韦治疗甲型和乙型流感病毒性肺炎的临床疗效

目的观察泉州清新方联合奥司他韦治疗甲型和乙型流感病毒性肺炎的疗效。方法选取2022年3月1日—2023年12月31日泉州市中医院收治的甲型和乙型流感病毒性肺炎患者146例,应用简单随机抽样法分为中西药联合组及奥司他韦组,每组73例。奥司他韦组应用磷酸奥司他韦治疗,中西药联合组在奥司他韦组治疗基础上应用泉州清新方治疗,2组均治疗5 d。比较2组临床疗效、临床症状缓解时间,治疗前后中医证候积分及不良反应。结果中西药联合组总有效率为91.78%,高于奥司他韦组的79.45%(χ^(2)=4.505,P=0.034);中西药联合组退热时间、咳嗽消失时间、咽痛消失时间及啰音消失时间均短于奥司他韦组(P<0.05);治疗5 d后,2组中医证候积分低于治疗前,且中西药联合组低于奥司他韦组(P<0.01);中西药联合组与奥司他韦组不良反应总发生率比较差异无统计学意义(12.33%vs.8.22%,χ^(2)=0.669,P=0.414)。结论泉州清新方联合奥司他韦可有效改善甲型和乙型流感病毒性肺炎的临床症状,缩短退热、咳嗽、咽痛及啰音消失时间,降低中医证候积分,未见明显不良反应。

俯卧位通气联合奥司他韦治疗甲型流感的效果

目的:分析俯卧位通气联合奥司他韦治疗甲型流感的效果。方法:选取2021年10月—2023年10月清华大学玉泉医院收治的甲型流感患者66例为研究对象,采取随机分组的方式分为对照组和观察组,各33例。对照组接受奥司他韦抗病毒治疗,观察组接受俯卧位通气联合奥司他韦抗病毒治疗。比较两组炎性反应指标、免疫功能指标、不良反应发生率、治疗效果、恢复情况。结果:治疗后,两组白细胞介素-10、C反应蛋白、白细胞计数水平低于治疗前,观察组低于对照组,差异有统计学意义(P<0.05);治疗后,两组CD3^(+)、CD4^(+)水平低于治疗前,CD8^(+)水平高于治疗前,观察组CD3^(+)、CD4^(+)水平低于对照组,CD8^(+)水平高于对照组,差异有统计学意义(P<0.05);两组不良反应发生率比较,差异无统计学意义(P>0.05);观察组治疗总有效率高于对照组,差异有统计学意义(P=0.031);观察组体温复常时间、呼吸道症状消失时间、住院时间短于对照组,差异有统计学意义(P<0.001)。结论:俯卧位通气联合奥司他韦治疗甲型流感的效果较好,可减轻炎性反应,提高免疫功能,促进症状恢复,缩短住院时间。

喉咽清口服液联合奥司他韦治疗儿童流行性感冒的效果及对HO-1、MIF和CysLTs水平的影响

目的探讨喉咽清口服液联合奥司他韦治疗儿童流行性感冒的效果及对血红素氧合酶-1(HO-1)、巨噬细胞迁移抑制因子(MIF)和半胱氨酰白三烯(CysLTs)水平的影响。方法选取2022年2月至2023年2月我院收治的100例流行性感冒患儿,以随机数字表法将其分为对照组(50例,奥司他韦)和观察组(50例,喉咽清口服液联合奥司他韦)。比较两组的治疗效果。结果观察组的治疗总有效率高于对照组,差异具有统计学意义(P<0.05)。观察组的发热消失时间、咳嗽消失时间、咽痛消失时间、流涕消失时间、乏力消失时间均短于对照组,差异具有统计学意义(P<0.05)。治疗后,观察组的HO-1水平高于对照组,MIF、CysLTs水平低于对照组,差异具有统计学意义(P<0.05)。治疗后,观察组的免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)水平均高于对照组,差异具有统计学意义(P<0.05)。结论喉咽清口服液联合奥司他韦治疗儿童流行性感冒的效果较好,可有效改善临床症状,减轻机体炎症反应,调节免疫功能,值得推广。