Antithrombin III(Grifols SA)

LACHEN, Switzerland, June 6, 2023 /PRNewswire/ -- Octapharma to present new clinical and scientific findings for wilate®, Nuwiq®, octaplex® and atenativ® at the upcoming ISTH 2023 Congress New data and clinical study developments will be presented in nine posters and during two Supported Symposia The most recent developments from Octapharma's Haematology and Critical Care portfolios will be presented at the 31st Congress of the International Society on Thrombosis and Haemostasis (ISTH) on June 24–28, 2023, in Montréal, Canada. The data will be featured in nine poster presentations and two Supported Symposia during this key international meeting. Octapharma is a proud Silver Supporter of the ISTH 2023 Congress. There will also be a promotional booth focusing on modern approaches to bleeding and coagulation management. People with inherited bleeding disorders, such as haemophilia A and von Willebrand disease, are at risk of recurrent and prolonged bleeds placing a heavy burden on patients' lives. In addition to the risk of potentially life-threatening bleeds, increased bleeding is associated with long-term consequences that severely impair patients' quality of life. Patients with a deficiency of the prothrombin complex of coagulation factors are also at increased risk of bleeding. Such deficiencies may be inherited but can also be acquired and may be induced by the use of vitamin K antagonists (VKA) for anticoagulation therapy. During urgent surgery, rapid correction of the deficiency is required and can be achieved through the use of four-factor prothrombin complex concentrates (4F-PCC). 4F-PCC can also be used to control major bleeding associated with the use of direct oral anticoagulant (DOAC) therapy in patients with factor Xa inhibitors. The presentations at ISTH 2023 reflect Octapharma's long-term commitment to improving patient care and addressing unmet clinical needs. "We are proud to showcase our new data and provide updates of studies from across Octapharma's haematology and critical care portfolios at ISTH 2023," said Olaf Walter, Board Member at Octapharma. "Despite advances in the field, people with bleeding and coagulation disorders still face considerable challenges surrounding bleed management. At Octapharma, we strive to address such challenges head-on with the goal to improve the day-to-day lives of patients." Poster Presentations The WIL-31 study investigated the efficacy and safety of prophylaxis with wilate® in patients with von Willebrand disease (VWD) of any gender with VWD type 1, type 2 (except 2N) or type 3. The results of the study show that wilate® prophylaxis is highly effective at reducing the number of bleeds in children and adults, and provides compelling evidence to support the use of regular prophylaxis in patients with severe VWD. Further data on the efficacy and safety of wilate® prophylaxis is being collected in the WIL-33 study in children under six years of age. The goals and design of this study will be presented. Prophylaxis is already the standard of care for people with haemophilia A. The results of an indirect comparison show that fewer patients experienced bleeds when receiving personalised prophylaxis with Nuwiq® rather than prophylaxis with a comparator factor VIII (FVIII) product. New data exploring differences in the binding of different recombinant FVIII products to platelets in vitro will be shared in a separate poster. Although haemophilia A mainly affects males, female carriers may also have reduced levels of FVIII and be at increased risk of bleeding, especially during traumatic events such as surgery. Data from a retrospective case series of female haemophilia A carriers who were prescribed Nuwiq® during surgery at the Bonn Haemophilia Centre will provide an insight into the effective treatment of females. MOTIVATE is an ongoing, international investigator-initiated study made possible through funding from Octapharma AG, evaluating the management of haemophilia A patients with inhibitors. The Coordinating investigators, Drs Carmen Escuriola Ettingshausen and Robert F. Sidonio Jr., will provide an update on the study status as a poster at the Congress. The LEX-210 study will evaluate the haemostatic efficacy of octaplex® in patients with acute major bleeding receiving factor Xa inhibitors. As anti-factor Xa (anti-Xa) activity levels will be measured in these patients, the correlation between heparin-calibrated and specific factor Xa inhibitor assays has been analysed and will be presented. The results from the first head-to-head, phase 3, randomised, controlled trial (LEX-209) comparing two 4F-PCCs in patients on VKA anticoagulation who need urgent surgery will be presented, with a focus on dosing and haemostatic efficacy results. The results show that octaplex® was non-inferior to control 4F-PCC for rapid vitamin K antagonist reversal in patients undergoing urgent surgeries with significant bleeding risk. The posters will be presented at the following times: Sunday June 25, 18:30–19:30 ET PB0163. Differential binding of recombinant FVIII products to platelets, and impact of FVIII on VWF expression and platelet contractility in vitro. Presenting author: Viola Vogel. PB0235. Phase 3 study of the efficacy, pharmacokinetics, immunogenicity and safety of von Willebrand factor/factor VIII concentrate in patients with severe von Willebrand disease under 6 years of age. Presenting author: Akshat Jain. PB0238. Prospective, phase III study of the efficacy, safety, and pharmacokinetics of a human antithrombin III concentrate in congenital antithrombin deficiency during surgery or childbirth. Presenting author: Craig Kessler. Monday June 26, 18:30–19:30 ET PB0840. Efficacy of prophylaxis with a plasma-derived von Willebrand factor/factor VIII concentrate in patients with von Willebrand disease: Subgroup analysis by age and VWD type. Presenting Author: Robert F. Sidonio Jr. PB0681. German experience with simoctocog alfa during surgery in female haemophilia A carriers. Presenting author: Natascha Marquart. PB0584. Using heparin-calibrated assays to assess anti-factor Xa activity of factor Xa inhibitors (FXaI): a literature correlation analysis. Presenting author: Ravindra Sarode. PB1032. Four-factor prothrombin complex concentrates for vitamin K antagonist reversal before urgent surgery: dosing and hemostatic efficacy in a double-blind randomized study. Presenting author: Ravindra Sarode. Tuesday June 27, 18:30–19:30 ET PB1261. Matching adjusted indirect comparisons between personalized prophylaxis with simoctocog alfa versus standard prophylaxis with turoctocog alfa in adults with severe haemophilia A. Presenting author: Fernando F. Corrales-Medina. PB1288. Immune tolerance induction (ITI) strategies in haemophilia A patients with inhibitors – data from the MOTIVATE study. Presenting author: Carmen Escuriola Ettingshausen. Wednesday June 28, 13:00–14:00 ET PO11. Impact of simoctocog alfa prophylaxis on bleeding and joint health in Spanish patients with severe haemophilia A. Presenting author: María Teresa Alvarez-Román. Supported Symposia New clinical and scientific data, as well as updates on ongoing and new studies, will be shared during two Supported Symposia during the Congress. Final Results of the WIL-31 Study – A New Treatment Standard for von Willebrand Disease Patients with Severe Bleeding? Monday June 26, 13:15–14:30 ET, Room 511 Chair: Alok Srivastava, Christian Medical College, Vellore, India The symposium will focus on new data regarding the use of wilate® prophylaxis in children and adults to improve the lives of people with VWD. Topics presented will include the importance of prophylaxis, including current guidelines on the management of VWD and how to identify patients who may benefit from prophylactic treatment. Key learnings from the WIL-31 VWD prophylaxis study will be shared and case presentations of select WIL-31 patient cases from across the globe will be discussed. Bringing Together Two Worlds – Scientific and Clinical Insights for All-Round Bleed Protection in Haemophilia A Tuesday June 27, 13:15–14:30 ET, Room 511 Chair: Davide Matino, McMaster University, Hamilton, Ontario, Canada Octapharma's symposium on haemophilia A this year is dedicated to both, scientists and clinicians. The speakers will present data from the NuPreviq trial and the matching adjusted indirect comparisons (MAIC) analyses of Nuwiq® vs extended half-life (EHL) products as well as introduce the PROVE study. Topics discussed during this symposium will also include the role of platelets in FVIII binding, impact of prophylaxis on microRNA expression (biomarkers of joint and bone health) and how to utilise the power of RNASeq to understand inhibitor development. Both symposia will be open to congress participants in Montréal and will be streamed live for participants not attending the Congress. The opportunity to ask questions will be available at both symposia. Larisa Belyanskaya, Head of IBU Haematology at Octapharma commented: "We are delighted to showcase new data from WIL-31, which supports the importance of prophylaxis in people with von Willebrand disease, and to discuss scientific and clinical questions that need to be address to advance the management of people with haemophilia A." About Octapharma Headquartered in Lachen, Switzerland, Octapharma is one of the largest human protein manufacturers in the world, developing and producing human proteins from human plasma and human cell lines. Octapharma employs more than11,000 people worldwide to support the treatment of patients in 118 countries with products across three therapeutic areas: Immunotherapy, Haematology and Critical Care. Octapharma has seven R&D sites and five state-of-the-art manufacturing facilities in Austria, France, Germany and Sweden, and operates more than 190 plasma donation centres across Europe and the US. Octapharma has 40 years of experience in patient care. About Nuwiq® Nuwiq® (simoctocog alfa) is a 4th generation recombinant factor VIII (rFVIII) protein, produced in a human cell line without chemical modification or fusion with any other protein[1]. It is cultured without additives of human or animal origin, is devoid of antigenic non-human protein epitopes and has a high affinity for von Willebrand factor[1]. Nuwiq® treatment has been assessed in nine[1-3] completed clinical trials which included 201 previously treated patients (190 individuals)[1] and 108 previously untreated patients[2] with severe haemophilia A. Nuwiq® is available in 250 IU, 500 IU, 1,000 IU, 1,500 IU, 2,000 IU, 2,500 IU, 3,000 IU and 4,000 IU presentations[4]. Nuwiq® is approved for use in the treatment and prophylaxis of bleeding in patients with haemophilia A (congenital FVIII deficiency) across all age groups[4]. About wilate ® wilate® is a high-purity human von Willebrand factor/factor VIII (VWF/FVIII) concentrate, that undergoes two virus inactivation steps during its production[5]. No albumin is added as a stabiliser[5]. The purification processes result in a 1:1 ratio of VWF to FVIII that is similar to normal plasma[5]. wilate® contains a VWF triplet structure and content of large high molecular weight multimers similar to normal human plasma[5]. wilate® is exclusively derived from large pools of human plasma collected in approved plasma donation centres[6]. wilate® is available in 500 IU and 1,000 IU presentations. wilate® is indicated for the prevention and treatment of haemorrhage or surgical bleeding in von Willebrand disease (VWD), when desmopressin (DDAVP) alone is ineffective or contra-indicated, and for the treatment and prophylaxis of bleeding in patients with haemophilia A (congenital factor VIII deficiency)[6]. About octaplex® octaplex® is a human four factor prothrombin complex concentrate (4F-PCC) containing Factors II, VII, IX and X, as well as inhibitor protein C and its cofactor protein S. octaplex® is indicated for the treatment of bleeding and perioperative prophylaxis of bleeding in acquired deficiency of the prothrombin complex coagulation factors, such as deficiency caused by treatment with vitamin K antagonists, or in case of overdose of vitamin K antagonists, when rapid correction of the deficiency is required. It can also be used in the treatment of bleeding and perioperative prophylaxis in congenital deficiency of the vitamin K dependent coagulation factors II and X when purified specific coagulation factor product is not available. octaplex® is presented in vials containing 500 IU and 1000 IU of human prothrombin complex. The total protein content per vial is 260–820 mg (500 IU vial)/ 520–1640 mg (1000 IU vial). The specific activity of the product is ≥ 0.6 IU/mg proteins, expressed as factor IX activity. Do not store above 25°C, do not freeze. Store in the original package in order to protect from light. The product shelf life is three years.[7] About atenativ® atenativ® is a human antithrombin III concentrate (AT III), which is presented in vials containing 500 IU and 1000 IU of human antithrombin III as lyophilised powder for infusion. After reconstitution with 10 mL water for injection (WFI), the solution provides 50 IU/mL or 100 IU/mL of human antithrombin III. Store in the refrigerator (2-8 °C) Keep the vials in the outer carton in order to protect from light. The shelf life is three years. It has been indicated for treatment of patients with congenital and acquired antithrombin deficiency including prophylaxis for the prevention of deep vein thrombosis and thromboembolism in clinical risk situations (especially in surgery, pregnancy and childbirth), in combination with heparin, if indicated, as well as prevention of the progression of deep venous thrombosis and thromboembolism associated with heparin as indicated.[8] atenativ® is currently not registered in Canada. 1. Lissitchkov T et al. Ther Adv Hematol 2019; 10:2040620719858471. 2. Liesner RJ et al. Thromb Haemost 2021; 121:1400-8. 3. Octapharma AG; Data on file. 4. Nuwiq® Summary of Product Characteristics. 5. Stadler M et al. Biologicals 2006; 34:281-8. 6. wilate® Summary of Product Characteristics. 7. octaplex® Summary of Product Characteristics. 8. atenativ® Summary of Product Characteristics. Octapharma press releases are specifically for health specialist/medical media and are not for consumer press. 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中文摘要目的    建立和验证全自动凝血分析仪检测人抗凝血酶Ⅲ（human antithrombin Ⅲ，AT-Ⅲ）活性的方法。方法    参照全自动凝血分析仪操作说明书进行AT-Ⅲ活性的检测，并对方法的线性、检测范围、精密度和准确度进行方法学验证。结果    全自动凝血分析仪检测AT-Ⅲ活性的方法稳定，仪器自动运行标准曲线的决定系数均>0.980，定标血浆标准曲线线性范围为0.12～1.23 IU/ml，定标血浆高、中、低值和定量下限稀释样品的效价回收率均在80%～120%范围内，且高、中、低值效价回收率的相对标准偏差（relative standard deviation，RSD）均<15%，定量下限稀释样品的效价回收率的RSD<20%，供试品的批内精密度RSD均<5%，批间精密度RSD均<10%，对3批AT-Ⅲ制品及其中间品进行活性检测，每批样品分别测定3次的平均RSD均＜5%。结论    全自动凝血分析仪检测AT-Ⅲ活性的方法具有较好的线性、检测范围、准确度和精密度，可用于AT-Ⅲ生产工艺中活性的检测。正文       人抗凝血酶Ⅲ（human antithrombin Ⅲ，AT-Ⅲ）是人体内一种重要的丝氨酸蛋白酶抑制剂，也是血液中最重要的抗凝血因子，主要抑制人血液中的凝血酶Ⅱa和凝血因子Ⅹa（factor Ⅹa，FⅩa）的活性[1-3]。AT-Ⅲ能与凝血酶或一些丝氨酸蛋白酶抑制剂按1∶1反应形成复合物，这种抗凝血酶-凝血酶复合物能够快速被肝脏细胞清除，在血液中的半衰期通常不超过5 min，可有效阻止血栓的形成。在肝素存在的情况下，AT-Ⅲ抑制凝血酶的活性增强1 000倍以上。AT-Ⅲ的主要生物学功能是抗凝血作用[4-7]。目前，AT-Ⅲ在国内还没有产品上市，因此中国药典2020年版尚未收录AT-Ⅲ的活性检测方法。本研究采用以AT-Ⅲ与肝素形成的复合物抑制凝血酶为基础的动力学方法，即AT-Ⅲ在适量肝素存在的情况下与FⅩa过量结合，剩余的FⅩa与FⅩa发色底物反应显色，检测波长405 nm处吸光度值（A405），建立AT-Ⅲ活性值与A450之间的回归曲线，依此检测人AT-Ⅲ生产工艺中AT-Ⅲ的活性，并对该检测方法的线性及检测范围、精密度和准确度进行方法学验证。1材料与方法1.1样品  AT-Ⅲ制品（批号201808004、201808005、201808006）、AT-Ⅲ原液（批号B20180608、B20180618、B20180627）、组分Ⅳ沉淀滤液（批号F20180608、F20180618、F20180627）、凝胶过滤亲和层析解离液（批号D20180608、D20180618、D20180627） 、肝素亲和层析解离液（批号H20180608、H20180618、H20180627）均由兰州生物制品研究所有限责任公司第四研究室制备。1.2主要试剂及仪器定标血浆（批号：315171，复溶后AT-Ⅲ标示效价为 0.92 IU/ml）购自美国Thermo公司，AT-Ⅲ测定试剂盒（批号：F1600679P1）购自法国HYPHEN BioMed公司，稀释液（批号：N1142958，规格：100 ml/瓶）、清洗液（批号：N0248417，规格：500 ml/瓶）、样品杯（批号：0005575100）、样品盘（批号：0006800000）均购自美国Instrumentation Laboratory公司；ACL7000全自动凝血分析仪购自美国贝克曼公司。1.3AT-Ⅲ活性检测方法的建立将稀释液放在全自动凝血分析仪样品盘的Dil位，依次将AT-Ⅲ测定试剂盒里的FⅩa及底物放在反应盘2和3位；复溶后的定标血浆放入样品盘的pool位，用于制定标准曲线；复溶后的待测AT-Ⅲ制品用稀释液稀释至1 IU/ml左右，放入样品盘的1—15位；按照AT-Ⅲ测定试剂盒说明书进行操作，记录每个样品的效价和A405。1.4方法验证由于AT-Ⅲ测定试剂盒规定的检测范围为0.10～1.50 IU/ml，因此将定标血浆分别用0.60、0.75、1.00、1.50、3.00及7.50 ml稀释液溶解，获得理论效价分别为1.53、1.23、0.92、0.61、0.31、0.12 IU/ml的稀释样品。1.4.1   线性    用1 ml注射用水充分溶解定标血浆，取0.5 ml放入样品盘的pool位，在1—12、17、18位放入14个空样品杯，将稀释液加入样品杯并置于16位，取4 ml稀释液加入4 ml样品杯并置于Dil位，运行仪器AT-Ⅲ检测程序下的CALIBRATION程序，设备自动将复溶的定标血浆再进行2倍系列稀释至3个浓度，将3个稀释度的A405和理论效价进行标准曲线的线性拟合。要求标准曲线的决定系数（R2）≥0.980。1.4.2   检测范围    将配制的6个稀释样品作为待测样品，进行样品检测程序，每个样品重复3次。将每个样品3次检测的A405均值与理论效价进行标准曲线的线性拟合，并将各个稀释样品A405均值代入该线性方程获得对应稀释样品的回算值，计算每个稀释样品的效价回收率和变异系数（coefficient of variation，CV），确认标准曲线的线性范围。效价回收率为回算值与理论效价的百分比。接受标准：标准曲线各稀释样品A405的CV≤10.0%；标准曲线的R2≥0.980，高、中、低值和正常值的效价回收率为理论效价的80%～120%，检测下限的效价回收率为理论效价的80%～120%。1.4.3 精密度1.4.3.1 批内精密度    将稀释后的3批AT-Ⅲ制品、稀释液、AT-Ⅲ活性测定试剂分别进行活性检测，每批样品重复测定9次，记录检测结果，并计算每批样品9次检测结果的相对标准偏差（relative standard deviation，RSD），接受标准：每批样品重复检测9次的RSD＜10%。1.4.3.2 批间精密度    3批AT-Ⅲ制品在3个工作日进行活性检测，每批次分别测定5次，根据每批次在3个工作日的试验结果计算RSD，接受标准：每批样品在不同时间内活性检测结果的RSD＜10%。1.4.4 准确度    将配制的高、中、低值和定量下限的定标血浆样品作为待测样品，每个样品5个复孔，计算效价回收率。接受标准：每个样品的效价回收率均为理论效价的80%～120%，高、中、低值效价回收率的RSD均＜15%，定量下限效价回收率的RSD＜20%。1.5方法的初步应用用建立的活性检测方法检测AT-Ⅲ制品和中间品各3批，每个样品重复检测3次，计算RSD，接受标准：每批样品活性检测结果的RSD＜5%。2结果2.1线性全自动凝血分析仪自动运行6次的标准曲线见表1。每个稀释度血浆浓度的CV均<10%；标准曲线的R2均> 0.980。2.2检测范围定标血浆不同稀释样品拟合曲线为一次线性方程y=1.028 5x-0.020 9，R2＝0.998，各稀释样品的回收率在95%～120%之间，标准曲线的线性范围在0.12～1.23 IU/ml之间，见表2。2.3精密度2.3.1  批内精密度    3批AT-Ⅲ制品分别重复测定9次的RSD均＜5%，见表3。2.3.2  批间精密度    3批 AT-Ⅲ制品在3个工作日进行活性检测，每批测定5次，平均RSD分别为7.35%、7.69%、9.64%，见表4。2.4准确度定标血浆高、中、低值和定量下限的效价回收率均为理论效价的80%～120%，其中高、中、低值效价回收率的RSD均＜15%，定量下限效价回收率的RSD＜20%，见表5。2.5方法的初步应用对3批AT-Ⅲ制品及其中间品进行活性检测，测定3次的平均RSD均＜5%，见表6。3讨论目前，AT-Ⅲ活性的检测方法只有欧洲药典收录的手工法，其检测原理是发色底物法，但该方法的前期准备工作繁琐，要消耗大量的时间和人力[8-11]。本研究建立了利用全自动凝血分析仪检测AT-Ⅲ活性的方法，其检测原理采用的是发色底物法，通过向样品中加入过量的FⅩa检测AT-Ⅲ的活性。在检测过程中，仪器的性能确认所用样品为定标血浆，但由于AT-Ⅲ纯化过程中涉及各种缓冲液，其中某些组分可能对检测有干扰，另外，由于目前市场上无AT-Ⅲ制剂的国际标准品，仅能用血浆标准品的标准曲线检测纯化的AT-Ⅲ样品，因此，需对两者的检测一致性进行评判。本研究检测3批AT-Ⅲ制品，每批样品重复检测9次的RSD均＜5%，3批AT-Ⅲ制品在不同日期各重复检测5次，其RSD均＜10%，表明该方法具有良好的精密度；定标血浆高、中、低值及定量下限稀释品的效价回收率均在理论效价的80%～120%范围内，且高、中、低值效价回收率的RSD均＜15%，定量下限稀释品效价回收率的RSD＜20%，表明该方法的准确性良好；全自动凝血分析仪自动运行标准曲线的R2均>0.980，标准曲线的检测范围在0.12～1.23 IU/ml之间；对3批AT-Ⅲ制品及其中间品进行活性检测，每批样品分别测定3次的平均RSD均＜5%，表明该方法可用于AT-Ⅲ生产工艺中活性的检测。本研究建立的利用全自动凝血分析仪检测AT-Ⅲ活性的方法具有操作简便、快速、准确度和精密度高等优势，且不需要过多人工操作，减少了人为操作误差，弥补了手工法的不足，具有良好的应用前景。作者付艳丽  孔涛  郭玉婷  曹扬  梁凌宇兰州生物制品研究所有限责任公司第四研究室，甘肃省疫苗工程技术研究中心，兰州 730046通信作者：梁凌宇，Email：lyliang@vacmic.com引用本文：付艳丽, 孔涛, 郭玉婷, 等. 人抗凝血酶Ⅲ生产工艺中活性检测方法的建立及初步应用 [J]. 国际生物制品学杂志, 2022, 45(4): 187-190.  DOI：10.3760/cma.j.cn311962-20211111-00073