Sabin-inactivated poliovirus vaccine(Takeda)

点击上方图片链接可报名随着社会进步和人类健康需求的提高，人用疫苗研发及生产技术发展迅速，我国在疫苗研发技术和生产工艺的改进取得了长足的发展。分别阐述了灭活病毒性疫苗、多糖蛋白结合疫苗、基因工程疫苗规模化生产工艺和技术的国内外现状和未来发展趋势。✦✦疫苗在保障国民健康和国家生物安全方面发挥了巨大作用。我国自实施计划免疫以来，传染病的发病率和死亡率得到有效控制，成功消灭了天花、脊髓灰质炎和孕妇新生儿破伤风，常见儿童传染病如麻疹、腮腺炎、风疹、百日咳、白喉、破伤风、乙型性脑炎、流行性脑脊髓膜炎等疾病的发病率和死亡率下降幅度在95% 以上，乙肝感染降到1% 以下，具有巨大的社会效益和经济效益。我国疫苗产业虽已取得了长足的进展，但疫苗产业的自主创新能力、新产品转化、关键核心技术应用、规模化生产工艺和技术水平，同发达国家相比仍有一定差距。而且随着全球经济一体化和社会的迅速发展，抗生素滥用、大气污染等多种因素影响，传染病的多发和高发态势仍然没有得到根本改变，生物恐怖的危害也不断加剧，一些重大疾病及新发、突发传染病，如艾滋病、流感、登革热、手足口病、埃博拉等仍严重威胁着人类生命健康。Part1国内外灭活病毒性疫苗的生产工艺与技术灭活病毒性疫苗为通过化学、热处理或辐射处理经培养纯化的病毒性疫苗，其主要优点是：与活病毒疫苗相比更稳定和安全，不会发生因疫苗接种而导致的感染；被灭活但仍可被机体的免疫系统识别，并产生有效的免疫应答；可用于免疫系统相对较弱的个体；稳定性较好，无需冷冻，方便运输。其缺点主要为免疫原性相对较弱，一般需进行多次接种。目前已经上市的灭活病毒性疫苗主要有脊髓灰质炎灭活疫苗、甲型肝炎灭活疫苗、流感病毒灭活疫苗、人用狂犬灭活疫苗、乙型脑炎灭活疫苗、双价肾出血热灭活疫苗（表1）[1]。表1　已上市灭活病毒性疫苗1.1细胞基质和培养方式目前国内外灭活病毒性疫苗的细胞基质主要为人二倍体细胞、原代细胞、鸡胚细胞和Vero细胞。其中人二倍体细胞主要有2BS 株、KMB-17 株、WI-38 株和MRC-5 株。各种细胞的特性不一样，培养方式也不一样。人二倍体细胞传代比例和传代代次比较受限，不利于大规模细胞培养，且对培养基和牛血清质量要求较高。原代细胞主要有地鼠肾细胞、沙鼠肾细胞，操作繁琐，存在外源因子污染风险。人二倍体细胞和原代细胞的培养方式主要为转瓶、细胞工厂，这两种培养方式较为传统，每批培养体积较小，较为费时费力，其生产规模仅仅是转瓶或细胞工厂数量上的增加，难以规模化放大。Vero细胞是第一个可连续传代的哺乳动物细胞，来源于非洲绿猴肾细胞，目前广泛用于疫苗的生产，为灭活病毒性疫苗常用的细胞基质。国内外主要采用微载体生物反应器进行Vero细胞培养。与细胞工厂培养方式相比，微载体生物反应器培养具有明显的优点：可进行高密度培养；可保证细胞在规定细胞传代限定代次内进行分级放大，易于进行规模化放大。国外已有企业建立了在8 周内从1mL 安瓶到6000L 生物反应器的Vero细胞放大生产工艺，在整个规模化放大过程中Vero细胞的产量和活性并不降低[2]。表2 显示了不同反应器规模细胞培养的生长状态。从中可以看出，在反应器内进行15 ～ 6000L 规模化放大培养时，Vero细胞密度体现较好的一致性[2]。表2　不同规格生物反应器培养Vero细胞正是由于Vero细胞在病毒性疫苗生产中的应用，使得现代病毒性疫苗的生产进入工业化、规模化时代，生产过程更加可控，批间一致性更好。目前国内外主流的以Vero细胞为基质的疫苗生产均采用微载体生物反应器培养工艺。以Vero细胞为基质生产的灭活病毒性疫苗主要有脊髓灰质炎灭活疫苗、狂犬病灭活疫苗、日本乙型脑炎灭活疫苗，目前Vero细胞还被用于小儿轮状病毒活疫苗Rotateq1（Merck）和Rotarix1（葛兰素史克）[3]、H5N1大流感疫苗Preflucel1（Baxter）[4]、日本脑炎疫苗Ixiaro1（Intercell）的生产[2]。此外，还被批准用于天花活疫苗ACAM20001（Sanofi Pasteur）的生产[2]。Vero细胞等传代细胞培养均采用含血清培养，但含血清培养对于病毒性疫苗的下游纯化也有较高的要求，需要严格控制血清中BSA的含量、纯化的回收率等一些关键的指标，所以无血清细胞培养技术成为国内外细胞培养研究热点之一。Vero细胞的研究的另一个热点是进行细胞悬浮化高密度培养。与使用微载体不同，细胞进行悬浮化培养后，细胞的贴壁特性发生了改变，细胞不需要消化可直接传代，并且可以提高细胞的发酵密度。1.2生产工艺国内外的灭活病毒性疫苗均需要经过收获、澄清、浓缩、纯化、灭活等生产工艺。其中纯化主要采用柱层析或蔗糖密度梯度离心法。这两种方式各有优缺点：层析法便于线性放大，批间一致性较好，但是试剂成本较高；蔗糖密度梯度离心法纯化的抗原成分单一，可有效区分空心和实心病毒颗粒，但每批处理量较小，后期需要除糖处理。国内外上市的灭活病毒性疫苗多采用层析纯化工艺。目前病毒灭活主要采用β-丙内酯和甲醛两种灭活剂。β-丙内酯常温下是无色黏稠状液体，主要通过作用于病毒RNA从而达到灭活病毒的效果。目前上市的疫苗中采用β-丙内酯灭活的主要有狂犬疫苗和双价肾出血热灭活疫苗。在疫苗研究初期人们就已使用甲醛进行细菌和病毒性疫苗的灭活。甲醛对于病毒核酸和蛋白质都具有破坏作用。甲醛对单链核酸的破坏最为有效，因此常被用于RNA 病毒的灭活。脊髓灰质炎灭活疫苗、甲型肝炎灭活疫苗、流感病毒灭活疫苗、乙型脑炎灭活疫苗等均采用甲醛灭活工艺。1.3质量控制灭活病毒性疫苗的主要控制指标为病毒抗原活性成分、纯度以及宿主细胞DNA、宿主细胞蛋白（HCP）、灭活剂等的残留成分控制。纯度的检测方法主要有HPLC、SDS-PAGE 或比活等指标。其中不同疫苗对Vero细胞残余DNA 含量要求不太统一，WHO生物学标准化专业委员会评估残留DNA引起的转化事件的风险后认为注射产品的DNA含量每剂小于10ng均可以接受[5]。目前，国内的狂犬疫苗等中的Vero细胞残余DNA含量标准为每剂不高于100pg。但美国上市的IPV（IPOL®，Sanofi Pasteur）Vero细胞残余DNA含量每剂低于10pg。国内上市的sIPV DNA含量为每剂不高于50pg。Vero细胞HCP含量一般为每剂不高于50ng。Part2多糖蛋白结合疫苗规模化生产工艺与技术2.1国外行业现状2.1.1　肺炎球菌结合疫苗目前，全球已上市的肺炎球菌结合疫苗有3 种，分别是惠氏（Wyeth）公司生产的7 价肺炎球菌结合疫苗Prevnar7（2000 年上市）和13 价肺炎球菌结合疫苗Prevnar13（2010 年上市），以及葛兰素史克公司生产的11 价肺炎球菌结合疫苗Synfl orix（2009 年上市）[6-8]。（1）PrevnarPrevnar7 配方中包括了4、6B、9V、14、18C、19F、23F 共7 个血清型的结合物，Prevnar13 包括了1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、23F 共13 个血清型的结合物，比Prevnar7 多6 个型别的结合物（1、3、5、6A、7F、19A），两者均以CRM197 为载体蛋白，均为磷酸铝佐剂吸附剂型。惠氏公司结合物制备采用的工艺路线为：将天然多糖进行酸水解降低分子量至适宜范围（有些型多糖在氧化反应中自行降解，不需要预先进行水解），用高碘酸钠氧化多糖邻羟基使多糖重复单位产生醛基，醛基与CRM197 分子上的氨基，在还原剂（如NaBH4）存在的条件下发生还原胺化反应生成酰胺键，该方法被称为还原胺化法。这种方法的优点是对多糖重复单位上的邻羟基进行可控的修饰，不破坏多糖分子的其他结构，易于质控。缺点是还原胺化反应需要的时间较长，有的型结合反应需要5d。在0.5mL Prevnar13 中，有12 种血清型多糖各2μg，4μg 6B型多糖，0.125mg 磷酸铝佐剂。Prevnar13的剂型为不含胶乳的单剂预充式注射器灌装注射液[9-10]。（2）Synfl orixSynfl orix 配方包括1、4、5、6B、7F、9V、14、18C、19F、23F 共10 个血清型结合物。该产品使用了3 种不同的载体蛋白，其中1、4、5、6B、7F、9V、14、23F 载体蛋白为不可分型流感嗜血杆菌表面蛋白D（PD），19F 载体蛋白为白喉类毒素（DT），18C 载体蛋白为破伤风类毒素（TT）。剂型同样为磷酸铝佐剂吸附剂型（0.5mg 磷酸铝佐剂/ 剂）。每剂疫苗含1、5、6B、7F、9V、14、23F 这7 个血清型多糖各1μg，4、18C、19F 各3μg。所采用的技术路线为对除5、6B、23F 这3 个型外的其他天然多糖分子进行微流化（microfl uidization）处理以降低分子量，在不同条件下用1- 氰基-4-（二甲氨基）吡啶四氟硼酸盐［1-cyano-4-（dimethylamino）pyridinium tetrafl uo-roborate，CDAP］将多糖的羟基活化为氰酸酯，活化后的多糖的氰酸酯通过与载体蛋白或己二酸二酰肼（adipicdihydazide，ADH）衍生的载体蛋白的氨基或肼基形成异脲衍生物而实现结合。其原理与传统的溴化氰活化法相同，但活化条件更加温和，活化物副反应发生率低，对多糖的结构改变较少。与还原胺化法相比，耗时少。此方法的缺点是活化时间仅几十秒，要在这几十秒内对溶液迅速变化的pH 进行控制并不容易，由于多糖活化和结合是连续完成的，难以对多糖活化进行有效质控，反应条件偏碱性，对多糖上碱性条件敏感的基团难免会造成破坏，结合载体蛋白利用率不高[11]。2.1.2　脑膜炎球菌多价结合疫苗当前国际上脑膜炎球菌结合疫苗的发展趋势为包含A、C、Y、W135 等4 个血清群的多价结合疫苗，目前全球已上市的有3 家产品，分别为赛诺菲巴斯德的Menactra、诺华的Menveo（2014 年被葛兰素史克收购）、葛兰素史克的Nimenrix。此外，还有一种联合疫苗，葛兰素史克的C、Y 群脑膜炎球菌与b 型流感嗜血杆菌结合疫苗，其产品名为Menhibrix。（1）Menactra液体剂型，2005年获得批准上市，规格为0.5mL，其中含A、C、Y、W135群脑膜炎球菌多糖各4μg，含载体蛋白白喉类毒素DT约48μg。其基本工艺路线为：①用弱酸和双氧水水解多糖，制备多糖水解物；②各群多糖水解物分别按照不同工艺进行衍生（A 群衍生物为多糖溶液中加入EDAC 和ADH，在A 群多糖的磷酸基团上连接肼基。C 群、Y 群、W135 群衍生物为利用氰基硼氢化钠将ADH 与多糖的末端醛基结合，衍生出肼基。然后多糖衍生物溶液加入EDAC，直接与DT 结合）。（2）Menveo2010 年获得批准上市。A群为冻干剂型，C、Y、W135 群为液体剂型，规格为0.5mL。每剂含A群多糖10μg，C、Y、W135 群多糖各5μg，载体蛋白为CRM197 蛋白。其基本工艺路线为：首先多糖用弱酸水解为分子量约为12 ～ 16 个重复单位的片段；在还原剂作用下，将多糖的末端醛基还原成氨基；然后经酯化产生末端活性酯基团；最后与CRM197 结合[12]。（3）Nimenrix2012 年获得批准上市，为冻干剂型，蔗糖作为赋形剂。每剂含A、C、Y、W135 群多糖各5μg，载体蛋白为破伤风类毒素。其基本工艺路线如下。①将荚膜多糖进行降解。②采用氰基活化法活化多糖。③ A 群和C 群结合：多糖用CDAP 活化后与ADH 进行衍生，然后与TT结合；④ Y 群和W135 群结合：用CDAP 活化后直接与TT 结合。（4）Menhibrix2012 年获得批准上市。为冻干剂型，赋形剂为蔗糖。每剂含b 型流感嗜血杆菌荚膜多糖2.5μg，C、Y 群多糖各5μg。载体蛋白为破伤风类毒素。每剂含12.6mg 蔗糖。2.2国内行业现状2.2.1　13 价肺炎球菌结合疫苗国内企业尚无上市的产品，当前进度领先的为玉溪沃森，已经完成Ⅲ期临床疫苗的接种工作，兰州生物正在进行Ⅱ期临床研究，民海生物也已进入了临床研究。2.2.2　AC 群脑膜炎球菌结合疫苗我国在20 世纪80 年代已开始研发A 群脑膜炎球菌结合疫苗（TT 为载体），上海生物制品研究所采用的先将蛋白质衍生后与多糖偶联的工艺，动物实验表明该法可增强结合物中多糖的免疫原性[13]，但研究未能进一步持续下去。后来国内多使用先将多糖衍生，再与蛋白质载体偶联的方法研制脑膜炎球菌结合疫苗。2006 年以来我国先后有4 家企业的AC 群脑膜炎球菌结合疫苗获准生产（表3）。这些产品都以TT 为载体，每剂每群多糖含量为10μg，以乳糖为保护剂；均采用氰基活化法进行多糖的衍生；结合物都采用柱层析纯化工艺。表3　我国已上市的AC 群脑膜炎球菌结合疫苗2.2.3　ACYW135 群脑膜炎球菌结合疫苗ACYW135 群脑膜炎球菌结合疫苗在国内尚未形成上市产品，据统计，国内已有6 家企业获得临床研究批件并进入临床阶段。进度最快的已经完成Ⅲ期临床研究，正在进行数据统计。Part3基因工程疫苗规模化生产工艺和技术基因工程疫苗是指使用DNA 重组技术，改造病原微生物的基因组，以降低其致病性，提高其免疫原性；或者将病原微生物基因组中的一个或多个保护性抗原编码的基因片段克隆到表达载体，通过原核或真核表达系统诱导表达外源蛋白，经纯化等工艺步骤制成疫苗，接种动物产生保护性抗体，达到防治传染病的目的。近年来，随着分子生物学、反相遗传学和结构生物学等理论研究和技术方法的不断突破，以及专业仪器的不断升级，基因工程疫苗的进展也日新月异。目前利用基因工程技术已经使用和正在研制开发的新型疫苗主要有基因工程亚单位疫苗、基因工程活载体疫苗、核酸疫苗、合成肽疫苗、转基因植物疫苗等，这些疫苗统称为基因工程疫苗。与传统疫苗相比，基因工程疫苗具有如下诸多优势：①抗原成分明确，表达通路清楚可控，安全性高；②生产成本低，工艺步骤稳定，易于规模化放大；③质量控制的方法和标准具有一定通用性，从而提高安全性；④通过载体或抗原蛋白的上游设计，可构建能够对多种疾病产生保护的多价疫苗。现在基因工程疫苗已成为生物制品产业发展的一种趋势，在各种疾病的防控方面发挥重要作用。基因工程疫苗的生产技术流程包括四个步骤：上游保护性抗原/ 表位和表达载体设计，基因工程菌/ 细胞构建，大规模发酵培养，目的蛋白纯化。基因工程蛋白的研发和规模化生产进入门槛高，尤其是建立工业规模生产平台，需要技术、人才和资本的多年积累。其中，高表达工程菌株/ 细胞株的构建和下游生产纯化工艺是关键技术[14]。以提高单位制品产量、细胞高密度培养及表达[15]、简化生产工艺、降低生产成本、保证大规模生产制品质量为目的的核心工艺开发已成为各大生物公司竞相开展的前沿课题。3.1国外基因工程疫苗规模化生产工艺与技术国际上一些知名疫苗制造企业在基因工程疫苗的制备方面已开始逐渐走向工艺规模化、自动化和一次性化制备阶段。工艺的规模化情况在一定程度上反映了制造工艺的先进性和稳定性。在大幅降低了疫苗产品成本的同时也降低了疫苗批间质量差异带来的风险。规模化培养是常规疫苗生产领域的主要技术，目前国际主流的培养规模为3000 ～8000L，多采用批式或批式加量培养，进行计算机二级控制，实现完全自动化[16]。近几年，针对不同宿主细胞特点和疫苗制品生产工艺，用于疫苗生产的个性化细胞培养基的需求不断增加，市场供求已趋于正规化。自动化是规模化生产的目标和趋势，即采用自动化的分离纯化体系，包括细胞破碎、微滤、超滤和层析等技术，同时，整体柱、移动模拟床色谱、膜色谱等新兴技术也逐渐应用于规模化生产中。自动化的优点在于生产过程中减少人力成本，提高效率，降低人为操作误差及污染风险。一次性产品包括规模化生产过程中接触制品的装置、器具和耗材，其应用可使制品质量可控、便于验证。一次性细胞培养反应器已发展为生物医药（包括疫苗）产品生产的主要形式，其发展对于降低疫苗生产成本、提高经济效益、确保制品的安全具有重要意义。国外疫苗生产企业多采用“个性化”服务的方式与某些企业合作建立一种“协议服务”的商业关系。通过模块化、平台化操作研制最适合自身产品特点的个性化技术应用于生产实践，厂家为客户提供最优化的技术服务以及相关的技术支持，例如定制培养基、定制层析介质等，帮助其提高产能，实现生产效率的大幅度提升[16]。规模性自动化、多参数系统化的疫苗生产技术平台为疫苗生产效率提高、生产工艺的一致性以及疫苗质量的稳定提供了保障。3.2国内基因工程疫苗规模化生产工艺与技术中国生物医药企业起步于20 世纪80 年代初期，经过30 年的努力，生物医药产业取得一定发展，生物产业已初具规模，在发展中国家属于较高水准。但我国疫苗的规模化生产工艺研究起步较晚，大多处于半自动化阶段，其根本原因在于，研发人员在“研究和生产的关联意识”和“质量源于设计”等理念上认识的差距，致使初期的研发设计与规模化生产制备脱节。临床前期的研究很少考虑到工艺的可放大性和终产品质量的可控性。制备工艺往往仅限于实验室规模，不适合规模化生产。如纯化工艺中，实验室研究阶段广泛的采用超速离心和亲和层析法，均为间断性操作且费用较高，很难应用于生产。随着国外疫苗研发和生产企业在国内广泛设厂投产及国内疫苗质量控制监察体系的不断完善，这方面差距在逐渐缩小。此外，疫苗生产用表达系统和宿主细胞方面，国外的研发机构通常拥有专利技术保护，具有相对成熟的技术储备；规模化生产的仪器设备方面，无论是高密度发酵的生物反应器还是规模化分离纯化的层析介质、膜分离技术[17]，几乎所有的关键仪器设备都依赖进口，这极大地限制了我国疫苗规模化发展的进程。基因工程疫苗领域重点在于上游设计，在疫苗研发初期综合考虑工艺放大可行性、质量可控性甚至厂房和设备需求，可迅速实现规模化生产占领市场，同时降低生产成本，提高生产效率，确保安全。因此，相比传统疫苗，我国的基因工程疫苗更容易在产业化能力方面与国际接轨。Part4未来发展趋势与展望4.1灭活病毒性疫苗技术和工艺的发展趋势灭活病毒性疫苗技术和工艺的发展趋势主要体现在以下几点。产业化与规模化程度进一步提高，微载体生物反应器规模更大、高密度发酵技术参数的优化等；新型细胞基质研究，进一步提高细胞基质的安全性和培养浓度；细胞和病毒培养基的优化，包括无动物来源的培养基配方研究等；纯化技术的进一步提高，最大限度提高目标抗原纯度和比活；利用反向遗传学等技术进行疫苗生产用毒株的改造。4.2多糖蛋白结合疫苗技术和工艺的发展趋势使用符合欧盟标准的原料多糖, 并在此基础上把原料多糖的C 多糖、残余蛋白和核酸含量控制在1% 以内。载体蛋白质使用精制后进一步纯化的TT，TT 单体含量达到90%，并使用基质辅助激光解吸- 飞行时间（MALDI-TOF）质谱技术检测蛋白质准确的分子量和单体含量。通过这些技术研发，使结合效率提高至30% ～ 40%，建立新的过程质量控制方法和临床效果评价方法，与国际接轨。4.3基因工程疫苗技术和工艺的发展趋势目前以基因工程制品为主导的基因工程应用产业已成为全球发展最快的产业之一，发展前景广阔。基因工程制品主要包括细胞因子、抗体、疫苗、激素和寡核苷酸药物等。它们对预防人类的肿瘤、心血管疾病、遗传病、糖尿病、包括艾滋病在内的各种传染病、类风湿疾病等有重要作用。在很多领域特别是疑难病症上，基因工程制品起到了传统化学药物难以达到的作用。中国生物医药市场将成为仅次于美国的全球第二大生物医药市场，预计在10 年创新发展的基础上，基因工程疫苗行业将迎来最佳时期。同时，基因工程疫苗的产业化也面临不小的挑战。由于在规模化方面中国长期以来以仿制为主，没有形成具有技术优势的领域，不能发挥技术的累积效应，制约了具有自主知识产权的产品的研发。因此， 面对国际新药研发的过程细化精准， 拥有自主知识产权，并依托研发产品的创新性平台技术是基因工程疫苗规模化生产的重中之重。平台技术的延续性和创新性将为更多基因工程疫苗的开发提供更广阔的发展空间。参考文献［1］ 普洛特金. 疫苗学[M]. 梁晓峰，罗凤基，封多佳，译. 北京：人民卫生出版社，2011.［2］ BARRETT P N，MUNDT W，KISTNER O，et al. 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BEIJING--(BUSINESS WIRE)-- Sinovac Biotech Ltd. (NASDAQ: SVA) (“SINOVAC” or the “Company”), a leading provider of biopharmaceutical products in China, has filed its 2022 annual report on Form 20-F with the U.S. Securities and Exchange Commission for the year ended December 31, 2022. The Company also reported its unaudited financial for the second half and audited financial for full year ended December 31, 2022. Second Half and Full Year 2022 Financial Summary Sales for the six months ended December 31, 2022 were $280.5 million, compared to $8.4 billion in the prior year period. Sales in 2022 were $1.5 billion, compared to $19.4 billion in the prior year. The Company posted $373.7 million of net loss attributable to common shareholders, or a loss of $3.76 per basic and diluted share, in the six months ended December 31, 2022, compared to net income attributable to common shareholders of $3.4 billion, or $33.79 per basic and $29.46 per diluted share, in the prior year period. The Company posted $107.9 million of net income attributable to common shareholders, or $1.08 per basic and $1.00 per diluted share in 2022, compared to net income attributable to common shareholders of $8.5 billion, or $85.20 per basic and $74.27 per diluted share, in the prior year period. Mr. Weidong Yin, Chairman, President and CEO of SINOVAC, commented “In 2022, we significantly increased our R&D investment and expanded our global market reach. Our R&D investment nearly tripled, and we are developing various vaccines to prevent over 20 life-threatening diseases. Growth was maintained in our non-covid business in 2022. We keep executing our business strategy of expanding our regular business into international market. More than 10 countries granted licenses to our non-COVID vaccines. In addition, our varicella vaccine and sIPV were successfully passed WHO prequalification assessment in 2022. “Looking ahead into 2023, we remain confident in our comprehensive product portfolio and growing business and scientific partnerships. Our mission to ‘supply vaccines to eliminate human diseases’ remains at the forefront, and we are committed to achieving this goal by investing in cutting-edge technology, expanding our global partnerships, and increasing accessibility to our life-saving vaccines,” Mr.Yin added. Business Updates COVID-19 Vaccine – CoronaVac®, the inactivated COVID-19 vaccine developed by SINOVAC, has been approved for use in more than 60 countries and regions worldwide. At this time, over 2.9 billion doses of CoronaVac® have been delivered globally, making SINOVAC the largest China-based COVID-19 vaccine provider to the international market. CoronaVac® has been validated by the World Health Organization ("WHO") for extended use in children as young as three years old under the Emergency Use Listing ("EUL") Procedure, in November 2022. This is the youngest age that the WHO has validated for EUL of COVID-19 vaccines in the world so far. CoronaVac® has also been fully registered in Hong Kong under the Pharmacy and Poisons Ordinance Cap 138 in December 2022, as one of the first COVID-19 vaccines approved for such official registration in Hong Kong. Since then, the vaccine can be supplied to registered medical practitioners and institutions for both private and public markets in Hong Kong. In 2022, the results of sufficient real-world studies and clinical trials in various regions and countries confirmed the safety and effectiveness of CoronaVac® for pediatric and adolescent populations, as well as demonstrated that the vaccine was effective in preventing multiple Omicron waves, particularly for individuals over the age of 60 who received three doses of the vaccine. Influenza Vaccines – Quadrivalent Influenza Vaccine, the latest development of SINOVAC's influenza vaccine family, entered the global market in 2022 by gaining overseas commercialization approvals. In 2022, the influenza vaccines became SINOVAC’s highest revenue generating non-COVID product for the first time. Recently, a new and state-of-the-art influenza vaccine production facility of SINOVAC started operations in Beijing. The plant, which complies with Chinese Good Manufacturing Practice (GMP) guidelines and utilizes green production processes, enables automated production at scale that expands SINOVAC capacity to meet the growing global demand for high-quality influenza vaccines. Varicella Vaccine – the live attenuated varicella vaccine was prequalified by the WHO in November 2022, marking the first WHO prequalified Chinese varicella vaccine, and received its first overseas order in 2022. Hepatitis A Vaccine – Healive®, the first and only WHO prequalified hepatitis A vaccine from China, has been registered in 24 countries and organizations worldwide thus far. Inactivated Enterovirus Type 71 (EV71) Vaccine – Inlive®, has been authorized for children between the ages of six months to three-years-old by BPOM, the food and drug agency of Indonesia, in November 2022, which is the first vaccine approved in Indonesia to protect from hand, foot and mouth diseases (HFMD). This is also Inlive®'s first overseas authorization. Sabin Inactivated Polio vaccine (“sIPV”) – the Poliomyelitis Vaccine (Vero Cell), Inactivated, Sabin Strains, was prequalified by WHO in June 2022, and sIPV is available for United Nations (UN) agencies to purchase to support the global polio eradication strategy. Quality Control – Two quality control laboratories in SINOVAC have been accredited by the China National Accreditation Service for Conformity Assessment (CNAS) in 2022. Unaudited Financial Results for the Second Half of 2022 Sales for the second half of 2022 were $280.5 million, compared to $8.4 billion in the prior year period. The decrease was mainly due to decreased sales of CoronaVac®. Selling, general and administrative expenses in the second half of 2022 were $667.7 million, compared to $428.5 million in the prior year period. R&D expenses in the second half of 2022 were $257.7 million, compared to $101.1 million in the prior year period. Net loss in the second half of 2022 was $702.3 million, compared to net income of $5.9 billion in the prior year period. Net loss attributable to common shareholders was $373.7 million, or a loss of $3.76 per basic and diluted share, in the second half of 2022, compared to a net income attributable to common shareholders of $3.4 billion, or $33.79 per basic and $29.46 per diluted share, in the prior year period. As the Company announced on February 22, 2019, the Company’s Board of Directors determined that certain shareholders became acquiring persons, as defined in the Company’s rights agreement (“Rights Agreement”), under which a trigger event occurred. As a result, the Company issued new common and preferred shares of SINOVAC. Without the effect of implementing the Rights Agreement and newly-issued common and preferred shares, basic and diluted loss per share for the second half of 2022 would be $5.17. Non-GAAP adjusted EBITDA was $1.0 billion loss in the second half of 2022, compared to $7.5 billion in the prior year period. Non-GAAP net loss was $805.8 million in the second half of 2022, compared to $5.9 billion net income in the prior year period. Non-GAAP diluted loss per share in the second half of 2022 was $3.79 compared to an earnings per share of $29.87 in the prior year period. Non-GAAP diluted loss per share in the second half of 2022, excluding the implementation of the Rights Agreement and the newly-issued common and preferred shares, would be $6.02. Reconciliations of non-GAAP measures to the nearest comparable GAAP measures are included at the end of this earnings announcement. The Company’s second half of 2022 financial statements are prepared and presented in accordance with U.S. GAAP. However, they have not been audited or reviewed by the Company’s independent registered accounting firm. Financial Results for the Twelve Months Ended December 31, 2022 Sales in 2022 were $1.5 billion, a decrease from $19.4 billion in the prior year. The decrease was due to decreased sales of CoronaVac®. Selling, general and administrative expenses in 2022 were $823.5 million, compared to $591.2 million in the prior year. R&D expenses in 2022 were $442.1 million, compared to $155.0 million in the prior year. The Company continued to invest in the advancement of pipeline vaccines. Net income in 2022 was $88.1 million, compared to $14.5 billion in the prior year. Net income decreased primarily due to decreased sales. Net income attributable to common shareholders was $107.9 million, or $1.08 per basic and $1.00 per diluted share, compared to net income attributable to common shareholders of $8.5 billion, or $85.20 per basic and $74.27 per diluted share, in the prior year. Excluding the implementation of the Rights Agreement, as described above, and the newly-issued common and preferred shares, basic and diluted earnings per share for 2022 would be $1.59. Non-GAAP adjusted EBITDA was $309.5 million loss in 2022, compared to income of $17.6 billion in the prior year. Non-GAAP net loss in 2022 was $177.0 million, compared to a net income of $14.5 billion in the prior year. Non-GAAP diluted loss per share in 2022 was $0.37, compared to an earnings of $74.67 per share in the prior year. Non-GAAP diluted loss per share in 2022, excluding the implementation of the Rights Agreement and the newly-issued common and preferred shares, would be $0.59 per share. Reconciliations of non-GAAP measures to the nearest comparable GAAP measures are included at the end of this earnings announcement. As of December 31, 2022, cash and cash equivalents and restricted cash totaled $4.3 billion, compared to $11.6 billion as of December 31, 2021. In 2022, net cash used in operating activities was $770.7 million, net cash used in investing activities was $5.8 billion, and net cash used in financing activities was $241.4 million. As of December 31, 2022, the Company had $0.3 million in bank loans due within one year. The Company expects that its current cash position will be able to support its operations for at least the next 12 months. Legal Proceedings As previously disclosed by the Company, on March 13, 2018, 1Globe Capital LLC (“1Globe”) filed a complaint against the Company in the Antigua Court. The trial of the matter took place from December 3 to 5, 2018. On December 19, 2018, the Antigua judge handed down his judgment (the “Antigua Judgment”), finding the Company fully in favor, dismissing 1Globe’s claim and declaring the Rights Agreement was validly adopted as a matter of Antigua law. On January 29, 2019, 1Globe filed a Notice of Appeal against the Antigua Judgment. On March 4, 2019, 1Globe filed an application for urgent interim relief, seeking an injunction to prevent the Company from continuing to implement its Rights Agreement until the resolution of the appeal. This application was heard on April 4, 2019, at which the Court of Appeal issued an order restraining the Company from operating the Rights Agreement in any way that affects 1Globe’s rights or shareholding or otherwise distributing the exchange shares to the Company’s shareholders who did not trigger the Rights Plan until after the determination of the appeal (the “Exchange Shares”). 1Globe’s appeal against the Antigua Judgment was heard on September 18, 2019, and the appeal decision was announced by the Eastern Caribbean Supreme Court, Court of Appeal (the “Court of Appeal”) on December 9, 2021, upholding the Antigua Judgment in each point. 1Globe applied for leave to appeal to the Judicial Committee of the Privy Council (the “Privy Council”), and the hearing of the application was held on February 24, 2022, in which the Court of Appeal granted 1Globe leave to appeal to the Privy Council on certain grounds, although not including the challenge to the validity of the Rights Agreement. On April 19, 2022, 1Globe renewed its application directly to the Privy Council for leave to appeal on its ground of appeal concerning the validity of the Rights Agreement. On July 13, 2022, 1Globe filed its Notice of Appeal on those grounds on which the Court of Appeal had granted 1Globe leave to appeal. On September 16, 2022, 1Globe filed an application to the Privy Council seeking permission to amend its existing application for permission to appeal and its existing Notice of Appeal, and to seek permission to appeal on another ground rejected by the Court of Appeal concerning the exercise of the Antigua Court’s discretion. The Company responded on October 21, 2022. On February 15, 2023, the Privy Council made a procedural decision to allow amendment of its existing application for permission to appeal, and decided to deal with procedural and substantive issues together at the Final Hearing. 1Globe has not yet taken steps to list a substantive hearing before the Privy Council. The appeal outcome is therefore pending. As previously disclosed, on March 5, 2018, the Company filed a lawsuit in the Court of Chancery of the State of Delaware, seeking a determination on whether 1Globe, the Chiang Li Family, OrbiMed Advisors, LLC and certain other shareholders of the Company had triggered the Rights Agreement. On April 12, 2018, 1Globe filed an amended answer to the Company’s complaint, counterclaims and a third-party complaint against the Company and Mr. Weidong Yin, alleging, among other allegations, that the Rights Agreement is not valid. On March 6, 2019, the Delaware Chancery Court entered a status quo order, providing that the Company not distribute any of the Exchange Shares to the Company’s shareholders who did not trigger the Rights Plan until the final disposition of the pending Delaware litigation or further order of the Court. On April 8, 2019, the Delaware Chancery Court stated that the Delaware litigation was pending the final outcome of 1Globe’s appeal of the Antigua Judgment. Separately, Heng Ren Investments LP (“Heng Ren”) filed suits against SINOVAC and Weidong Yin on May 31, 2019 in Massachusetts state court for the alleged breach of fiduciary duties and wrongful equity dilution. SINOVAC moved the matter from the state court to the United States District Court for the District of Massachusetts. Subsequently, on April 29, 2021, Heng Ren filed an amended complaint which alleged that Mr. Yin breached fiduciary duties owed to minority shareholders, that SINOVAC aided and abetted breaches of fiduciary duties and that both SINOVAC and Mr. Yin engaged in wrongful equity dilution. Heng Ren requested damages, attorney fees, and prejudgment interest. In July 2021, SINOVAC moved to dismiss Heng Ren’s amended complaint in the federal court in Massachusetts. On March 4, 2022, the court granted the motion as to the breach of fiduciary duty claims and denied the motion as to the wrongful equity dilution claim, and denied reconsideration of its decision on the motion. SINOVAC has answered the complaint. On July 2, 2018, the Company completed a private placement of its common shares with two private investors (the “PIPE Transaction”). On December 5, 2022, a purported shareholder filed a putative class action complaint in the United States District Court for the District of Massachusetts, asserting a claim under Section 204 of the Antigua and Barbuda International Business Corporations Act related to the PIPE Transaction, alleging that all shareholders were harmed in an identical manner to one another by the PIPE Transaction because the shares that were issued in the PIPE Transaction allegedly undervalued SINOVAC and all shareholders were purportedly wrongfully diluted as a result. The purported shareholder is represented by the same attorney who represents Heng Ren, and requests damages, attorneys’ fees, and prejudgment interest. On January 18, 2023, the Company filed a motion to dismiss in such purported shareholder’s matter (the “Motion to Dismiss”). On February 15, 2023, the court stayed discovery in the Heng Ren matter pending the resolution of the Motion to Dismiss. The Motion to Dismiss was fully briefed as of March 9, 2023, and is currently pending before the court. Status of Exchange Shares and Trading in the Company’s Shares As a result of the pending legal proceedings described above, the Exchange Shares are expected to remain in a trust for the benefit of the Company’s shareholders who did not trigger the Rights Plan until, at least, the conclusion of the appeal against the Antigua Judgement and the final disposition of the Delaware litigation or further order of the Delaware Chancery Court. The Exchange Shares remain issued and outstanding. The Nasdaq Stock Market LLC implemented a halt on trading of the Company’s common shares at the time the Exchange Shares were issued to the trust. The Company is currently unable to estimate when trading will resume, or if Nasdaq will take any additional action in regards to trading of the Company’s common shares. About SINOVAC Sinovac Biotech Ltd. (SINOVAC) is a China-based biopharmaceutical company that focuses on the R&D, manufacturing, and commercialization of vaccines that protect against human infectious diseases. SINOVAC’s product portfolio includes vaccines against COVID-19, enterovirus 71 (EV71) infected hand-foot-mouth disease (HFMD), hepatitis A, varicella, influenza, poliomyelitis, pneumococcal disease, mumps, etc. The COVID-19 vaccine, CoronaVac®, has been approved for use in more than 60 countries and regions worldwide. The hepatitis A vaccine, Healive®, passed WHO prequalification requirements in 2017. The EV71 vaccine, Inlive®, is an innovative vaccine under "Category 1 Preventative Biological Products" and commercialized in China in 2016. In 2022, SINOVAC’s Sabin-strain inactivated polio vaccine (sIPV) and varicella vaccine were prequalified by the WHO. SINOVAC was the first company to be granted approval for its H1N1 influenza vaccine Panflu.1®, which has supplied the Chinese government's vaccination campaign and stockpiling program. The Company is also the only supplier of the H5N1 pandemic influenza vaccine, Panflu®, to the Chinese government stockpiling program. SINOVAC continually dedicates itself to new vaccine R&D, with more combination vaccine products in its pipeline, and constantly explores global market opportunities. SINOVAC plans to conduct more extensive and in-depth trade and cooperation with additional countries, and business and industry organizations. For more information, please visit the Company’s website at . Safe Harbor Statement This press release may include certain statements that are not descriptions of historical facts, but are forward-looking statements. These statements are made under the “safe harbor” provisions of the U.S. Private Securities Litigation Reform Act of 1995. These forward-looking statements can be identified by terminology such as “will,” “expects,” “anticipates,” “future,” “intends,” “plans,” “believes,” “estimates” and similar statements. Forward-looking statements involve risks, uncertainties and other factors that could cause actual results to differ materially from those contained in any such statements. In particular, the outcome of any litigation is uncertain, and the Company cannot predict the potential results of the litigation it filed or filed against it by others. Additionally, the triggering of a shareholder rights plan is nearly unprecedented, and the Company cannot predict the impact on the Company or its stock price as a result of the trigger of the rights plan. Non-GAAP Financial Measures To supplement its consolidated financial statements, which are prepared and presented in accordance with GAAP, SINOVAC uses the following non-GAAP financial measures: non-GAAP adjusted EBITDA, non-GAAP net income (loss) and non-GAAP diluted earnings (loss) per share. For more information on these non-GAAP financial measures, please refer to the table captioned “Reconciliations of non-GAAP Measures to the Nearest Comparable GAAP Measures” in this results announcement. SINOVAC believes that non-GAAP adjusted EBITDA, non-GAAP net income (loss) and non-GAAP diluted earnings (loss) per share help identify underlying trends in its business that could otherwise be distorted by the effect of certain income or expenses that SINOVAC includes in net income and diluted earnings (loss) per share. SINOVAC believes that non-GAAP adjusted EBITDA, non-GAAP net income (loss) and non-GAAP diluted earnings (loss) per share provide useful information about its core operating results, enhance the overall understanding of its past performance and future prospects and allow for greater visibility with respect to key metrics used by our management in its financial and operational decision-making. Non-GAAP adjusted EBITDA, non-GAAP net income (loss) and non-GAAP diluted earnings (loss) per share should not be considered in isolation or construed as an alternative to income from operations, net income (loss), diluted earnings (loss) per share, or any other measure of performance or as an indicator of SINOVAC’s operating performance. These non-GAAP financial measures presented here may not be comparable to similarly titled measures presented by other companies. Other companies may calculate similarly titled measures differently, limiting their usefulness as comparative measures to our data. Non-GAAP adjusted EBITDA represents net income (loss) and excludes interest and financing expenses, interest income, net other income and income tax benefit (expenses), and certain non-cash expenses, consisting of share-based compensation expenses, amortization and depreciation that SINOVAC does not believe are reflective of the core operating performance during the periods presented. Non-GAAP net income (loss) represents net income (loss) before share-based compensation expenses and foreign exchange gain or loss. Non-GAAP diluted earnings (loss) per share represents non-GAAP net income (loss) attributable to common shareholders divided by the weighted average number of shares outstanding during the periods on a diluted basis, including accounting for the effect of the assumed conversion of options. SINOVAC BIOTECH LTD. Consolidated Balance Sheets As of December 31, 2022 and 2021 (Expressed in thousands of U.S. Dollars) December 31, 2022 December 31, 2021 Current assets Cash and cash equivalents $ 4,278,124 $ 11,608,855 Restricted cash 8,253 10,905 Short-term investments 7,034,569 1,806,449 Accounts receivable - net 537,118 952,402 Inventories 180,719 375,511 Prepaid expenses and deposits 15,242 160,987 Income tax receivable 72,371 -- Total current assets 12,126,396 14,915,109 Property, plant and equipment – net 993,781 903,298 Prepaid land lease payments 69,815 39,730 Intangible assets - net 9,699 1,480 Long-term prepaid expenses 23 25 Long-term investments 661,440 655,835 Prepayments for acquisition of equipment 120,912 65,290 Deferred tax assets 71,118 52,031 Right-of-use assets 58,586 115,376 Other Non-current Assets 2,798 -- Total assets 14,114,568 16,748,174 Current liabilities Short-term bank loans and current portion of long-term bank loans 293 3,099 Loan from a non-controlling shareholder 4,358 1,582 Accounts payable and accrued liabilities 905,923 1,020,651 Income tax payable -- 1,267,504 Deferred revenue 17,955 79,941 Deferred government grants 15,120 12,559 Dividend payable 141,993 17,125 Lease liability 5,993 10,385 Total current liabilities 1,091,635 2,412,846 Deferred government grants 4,477 4,870 Long-term bank loans 11,513 12,668 Deferred tax liability 241,526 324,164 Loan from a non-controlling shareholder -- 4,708 Lease liability 52,516 112,465 Other non-current liabilities 240 444 Total long-term liabilities 310,272 459,319 Total liabilities 1,401,907 2,872,165 Commitments and contingencies Equity Preferred stock 15 15 Common stock 100 100 Additional paid-in capital 540,582 547,691 Subscriptions receivable -- (7,109) Accumulated other comprehensive income (loss) (383,276) 130,622 Statutory surplus reserves 1,538,013 1,514,297 Accumulated earnings 7,225,987 7,141,819 Total shareholders' equity 8,921,421 9,327,435 Non-controlling interests 3,791,240 4,548,574 Total equity 12,712,661 13,876,009 Total liabilities and equity $ 14,114,568 $ 16,748,174 SINOVAC BIOTECH LTD. Consolidated Statements of Comprehensive Income (Loss) For the six and twelve months ended 2022 and 2021 (Expressed in thousands of U.S. Dollars, except for numbers of shares and per share data) Six months ended December 31 Year ended December 31, 2022 2021 2022 2021 (Unaudited) (Unaudited) Sales $ 280,471 $ 8,393,848 $ 1,492,761 $ 19,374,904 Cost of sales 430,094 440,455 684,456 1,072,221 Gross profit (149,623) 7,953,393 808,305 18,302,683 Selling, general and administrative expenses 667,663 428,510 823,543 591,167 Provision for doubtful accounts 3,461 3,528 4,268 2,967 Research and development expenses 257,714 101,129 442,108 155,040 Loss on disposal of property, plant and equipment 1,927 683 5,213 977 Government grants recognized in income (746) (39) (760) (725) Total operating expenses 930,019 533,811 1,274,372 749,426 Operating income (loss) (1,079,642) 7,419,582 (466,067) 17,553,257 Interest and financing expenses (641) (1,314) (1,264) (2,836) Interest income 106,820 71,764 190,818 102,568 Other income (expense), net 72,293 (88,999) 301,751 (89,948) Income (loss) before income taxes (901,170) 7,401,033 25,238 17,563,041 Income tax benefit (expense) 198,855 (1,534,196) 62,893 (3,104,130) Net income (loss) (702,315) 5,866,837 88,131 14,458,911 Less: (income) loss attributable to non-controlling interests 331,669 (2,507,683) 25,735 (5,991,431) Net income (loss) attributable to shareholders of Sinovac (370,646) 3,359,154 113,866 8,467,480 Preferred stock dividends (3,024) (3,024) (5,982) (5,982) Net income (loss) attributable to common shareholders of Sinovac (373,670) 3,356,130 107,884 8,461,498 Net income (loss) (702,315) 5,866,837 88,131 14,458,911 Other comprehensive income (loss), net of tax of nil Foreign currency translation adjustments (279,794) 162,103 (859,045) 193,098 Comprehensive income (loss) (982,109) 6,028,940 (770,914) 14,652,009 Less: comprehensive (income) loss attributable to non-controlling interests 445,656 (2,578,813) 370,882 (6,073,832) Comprehensive income (loss) attributable to shareholders of Sinovac (536,453) 3,450,127 (400,032) 8,578,177 Earnings (loss) per share Basic net income (loss) per share (3.76) 33.79 1.08 85.20 Diluted net income (loss) per share (3.76) 29.46 1.00 74.27 Weighted average number of shares of common stock outstanding Basic 99,502,243 99,328,085 99,502,243 99,311,551 Diluted 114,172,782 114,022,517 114,172,782 114,005,983 SINOVAC BIOTECH LTD. Consolidated Statements of Cash Flows For the six and twelve months ended 2022 and 2021 (Expressed in thousands of U.S. Dollars) Six months ended December 31 Year ended December 31, 2022 2021 2022 2021 (Unaudited) (Unaudited) Operating activities Net income (loss) $ (702,315) $ 5,866,837 $ 88,131 $ 14,458,911 Adjustments to reconcile net income to net cash provided by (used in) operating activities: Deferred income taxes (67,172) 335,990 (83,010) 265,096 Share-based compensation -- 6,223 -- 7,735 Inventory provision 83,800 68,700 140,004 70,133 Provision for doubtful accounts 3,461 3,528 4,268 2,967 Loss on disposal of property, plant and equipment 1,927 683 5,213 977 Depreciation of property, plant and equipment 78,660 67,920 153,819 84,446 Amortization of prepaid land lease payments 1,374 831 1,749 2,203 Amortization of intangible assets 932 94 1,029 183 Government grants recognized in income (746) (39) (760) (725) Changes in: Accounts receivable 429,615 423,635 289,600 (710,355) Inventories (6,869) (32,532) 30,382 (334,062) Income tax payable (191,672) (111,080) (1,275,296) 1,216,717 Prepaid expenses and deposits (111,395) (100,859) 22,519 (143,324) Deferred revenue (13,364) (255,923) (58,485) (288,779) Accounts payable and accrued liabilities 536,063 (8,173) (86,860) 720,858 Other non-current liabilities (5,531) (431) (3,048) (440) Net cash provided by (used in) operating activities 36,768 6,265,404 (770,745) 15,352,541 Financing activities Proceeds from bank loans 1 165 151 13,511 Repayments of bank loans (48) (19,534) (2,981) (33,436) Proceeds from issuance of common stock, net of share issuance costs -- 996 -- 1,033 Dividend paid (260,702) (1,838,094) (263,171) (1,875,892) Proceeds from subsidiary's financing -- -- 11,291 -- Government grants received 9,776 3,082 14,797 4,317 Loan from a non-controlling shareholder -- 9 -- 7,119 Repayments of loan from a non-controlling shareholder (1,474) (13,307) (1,486) (13,332) Net cash used in financing activities (252,447) (1,866,683) (241,399) (1,896,680) Investing activities Purchase of investments (5,717,991) (4,446,838) (11,658,465) (8,826,611) Proceeds from redemption of investments 3,619,956 5,018,775 6,327,521 6,567,491 Proceeds from disposal of equipment 5 120 608 172 Prepaid land lease payments (26,025) (18,596) (35,683) (30,986) Acquisition of intangible assets (9,568) (154) (9,568) (154) Purchase of equity investments (42,155) (13,663) (105,435) (13,663) Purchase of property, plant and equipment (114,402) (288,338) (393,800) (719,823) Proceeds from dissolution of subsidiary 114,352 -- 114,352 -- Net cash provided by (used in) investing activities (2,175,828) 251,306 (5,760,470) (3,023,574) Effect of exchange rate changes on cash and cash equivalents and restricted cash (189,210) 113,373 (560,769) 137,269 Increase (decrease) in cash and cash equivalents and restricted cash (2,580,717) 4,763,400 (7,333,383) 10,569,556 Cash and cash equivalents and restricted cash, beginning of period 6,867,094 6,856,360 11,619,760 1,050,204 Cash and cash equivalents and restricted cash, end of period 4,286,377 11,619,760 4,286,377 11,619,760 SINOVAC BIOTECH LTD. Reconciliations of Non-GAAP measures to the nearest comparable GAAP measures For the six and twelve months ended 2022 and 2021 (Expressed in thousands of U.S. Dollars, except for numbers of shares and per share data) Six months ended December 31 Year ended December 31, 2022 2021 2022 2021 (Unaudited) (Unaudited) (Unaudited) (Unaudited) Net income (loss) $ (702,315) $ 5,866,837 $ 88,131 $ 14,458,911 Adjustments: Share-based compensation -- 6,223 -- 7,735 Depreciation and amortization 80,966 68,845 156,597 86,832 Interest and financing expenses, net of interest income (106,179) (70,450) (189,554) (99,732) Other expense (income), net (72,293) 88,999 (301,751) 89,948 Income tax expense (198,855) 1,534,196 (62,893) 3,104,130 Non-GAAP adjusted EBITDA (998,676) 7,494,650 (309,470) 17,647,824 Net income (loss) (702,315) 5,866,837 88,131 14,458,911 Add: Foreign exchange (gain) loss (103,472) 73,179 (265,091) 68,026 Add: Share-based compensation -- 6,223 -- 7,735 Non-GAAP net income (loss) (805,787) 5,946,239 (176,960) 14,534,672 Net income (loss) attributable to common shareholders of Sinovac (373,670) 3,356,130 107,884 8,461,498 Add: Preferred stock dividends 3,024 3,024 5,982 5,982 Net income (loss) attributable to common shareholders of Sinovac for computing diluted earnings per share (370,646) 3,359,154 113,866 8,467,480 Add: Non-GAAP adjustments to net income (61,239) 46,868 (156,332) 45,038 Non-GAAP net income (loss) attributable to common shareholders of Sinovac for computing non-GAAP diluted earnings per share (431,885) 3,406,022 (42,466) 8,512,518 Weighted average number of shares on a diluted basis 114,172,782 114,022,517 114,172,782 114,005,983 Diluted earnings (loss) per share (3.76) 29.46 1.00 74.27 Add: Non-GAAP adjustments to net income (loss) per share (0.54) 0.41 (1.37) 0.40 Non-GAAP diluted earnings per share (4.30) 29.87 (0.37) 74.67 View source version on businesswire.com: Contacts Sinovac Biotech Ltd. Helen Yang Tel: +86-10-8279 9720 Email: ir@sinovac.com Source: Sinovac Biotech Ltd. View this news release online at: