新发传染病相关的药品技术战略储备思考

目的： 对新发传染病相关的药品技术战略储备进行探讨,旨在为战胜此次新冠肺炎疫情,更为下一次可能的疫情做好准备。方法： 梳理了世界卫生组织、欧盟、美国对与公共健康威胁相关的药品技术储备情况,主要阐述了新发传染病相关的疫苗技术储备信息,给出WHO建议的新型冠状病毒肺炎（COVID-19）药品研发信息,EMA在公共健康威胁（例如大流行性流感）相关药品方面的技术储备情况,并总结了EMA对大流行防备疫苗的技术要求。结果与结论： 建议在我国建立由国家主导并投资的与公共健康威胁相关的预防用疫苗或治疗用药物的技术和产品战略储备,例如为新发传染病的疫苗开发制定清晰的计划、法规和程序。药品技术和产品的战略储备可包括：1）政府部门制定与公共健康威胁相关疾病的清单,例如可能引起重大流行病的突发疾病用药优先研发清单;2）审评审批方面可考虑制定大流行性疾病疫苗（包括大流行防备疫苗）、大流行性疾病治疗药物研发的技术要求,涉及质量、非临床、临床、申报程序等内容;3）通过国家科技立项等方式投资预防用疫苗及治疗用药物的研制;4）引导生产企业进行相关的药品战略储备。     

过热蒸汽灭菌松花粉的安全性毒理学评价研究

目的 对过热蒸汽灭菌松花粉进行安全性毒理学评价,为其食用安全性提供科学依据。方法 根据国家标准要求和相关规定,对其进行急性经口毒性试验、细菌回复突变试验、哺乳动物红细胞微核试验、小鼠精子畸形试验和28天经口毒性试验等毒理学评价试验。结果 受试物最大给予量达10 g/(kg·BW)时,对小鼠未见毒性作用;细菌回复突变试验、哺乳动物红细胞微核试验和小鼠精子畸形试验结果均为阴性,未见该样品有致突变作用;28天经口毒性试验各项指标均未见明显毒性反应。结论 过热蒸汽灭菌松花粉的急性毒性分级属于实际无毒级,无遗传毒性,在该试验研究剂量和条件下,未见明显毒性作用,具有较高的安全性。     

基于产品生命周期的疫苗质量风险管理研究

目的：明确产品生命周期各阶段质量风险管理重点工作及控制措施,为产品全生命周期质量风险管理提供有力保障。方法：以GMP、PIC/s GMP附件20、ICH Q8、ICH Q9、ICH Q10、ICH Q12、WHO质量风险管理指南为基础,运用归纳总结及分析评估方法,对疫苗产品生命周期各阶段的质量风险管理进行研究。结果：通过质量风险管理研究,明确了生命周期不同阶段影响疫苗质量的关键因素,以满足科学和法规符合性为基础,提出具体控制措施,有效防控质量风险。结论：质量风险管理伴随疫苗产品全生命周期,任何一个阶段的质量风险管理工作都需重视并认真执行,采取科学合规的控制措施,可达到有效防控质量风险的目的。     

mRNA脂质纳米粒载药系统的构建及体外评价

目的 构建脂质纳米粒DLin-LNP,以EGFP-mRNA为模型药,考察DLin-LNP对于mRNA的体外递送能力。方法 采用薄膜水化法制备DLin-LNP, 并进一步制备 DLin@mRNA,对纳米粒进行表征,使用激光扫描共聚焦显微镜观察脂质纳米粒胞内的分布情况,以RM-1细胞为模型考察胞内转染情况。结果 成功制备了脂质纳米粒DLin-LNP,其粒径为（151.1±2.1） nm,空载电位为（23.7±0.5） mV。DLin-LNP在RM-1细胞中转染mRNA效率较高,其毒性远低于市售脂质体Lipo8000_,且 DLin-LNP脂质纳米粒稳定性好。结论 DLin-LNP具有高转染效率和安全性,且稳定性好,可作为mRNA递送载体,为后续脂质纳米粒肿瘤治疗中的应用提供依据。     

GLP体系下对实验动物供应商审核的要求

目的：规范实验动物供应商管理。方法：依据《药物非临床研究质量管理规范》（GLP）等相关法规要求,对实验动物供应商审核相关流程和要求进行了论述。结果与结论：实验动物在药物安全性评价中占有极其重要的地位。通过规范实验动物供应商审核流程,加强动物来源控制,可以有效地提高药物非临床原始资料的有效性和真实性,充分发挥GLP体系下实验动物供应商审核的优越性,保证非临床研究水平。     

以部分疫苗为例分析我国预防用生物制品的改良情况

目的：以百日咳疫苗、人用狂犬病疫苗的改良情况为例,分析监管政策对预防用生物制品技术改良的影响。方法：通过数据分析和文献研究,对我国已批准疫苗的技术改良情况进行分析。结果：技术改良是促进疫苗产业发展的重要手段。结论：监管机构需要通过政策的引导和鼓励促进疫苗的技术改良。     

纳米氧化锌安全性评价及化妆品法规管理现状

目的：了解国内外纳米氧化锌安全性评价及化妆品法规管理现状,为我国对纳米原料与纳米化妆品监管提供参考。方法：通过文献研究,总结目前国内外纳米氧化锌的毒理学研究进展及世界主要国家及地区对纳米氧化锌的化妆品法规管理现状。结果：透皮吸收试验发现防晒霜中的纳米氧化锌主要聚集在人体皮肤角质层和毛囊皮脂腺开口处,微量锌可经皮肤吸收进入血液,但不能确定是以氧化锌还是锌离子的形式吸收。体外彗星试验结果显示纳米氧化锌引起人表皮细胞、人鼻粘膜细胞DNA损伤。亚急性经口毒性试验发现纳米氧化锌通过氧化应激介导小鼠肝脏细胞DNA损伤和凋亡。纳米氧化锌对小鼠具有生殖发育毒性。急性吸入毒性试验显示纳米氧化锌导致大鼠肺脏、肝脏组织损伤。亚慢性毒性试验显示纳米氧化锌高剂量组（536.8 mg·kg^-1）SD大鼠出现贫血及轻度到中度胰腺炎,纳米氧化锌的未观察到有害作用水平（NOAEL）为268.4 mg·kg^-1。欧盟、美国、中国台湾已出台了一系列化妆品纳米材料的法规或文件,指导化妆品行业开展纳米材料的安全性评价。结论：化妆品中纳米氧化锌对人体具有潜在安全风险,我国应尽快制定化妆品中纳米原料的相关管理政策。     

组分百日咳疫苗安全性检测体外替代方法的应用和初步评价

目的：用一种组分百日咳疫苗残留毒性的体外替代检测方法，更稳定和客观地评价疫苗成品的生产一致性。方法：验证直接定性法和间接定量法2种中华仓鼠卵巢细胞簇集试验方法，分别对6个国内外厂家的10种组分百日咳疫苗产品毒性进行定量和定性检测，并使用百日咳毒素参考品将体外法与小鼠组胺致敏试验进行桥接和初步评价。结果：体外定量试验的灵敏度为0.0026±0.0003 IU·mL^-1，几何变异系数为13%，体外定性试验的灵敏度为0.0067±0.0016 IU·mL^-1，几何变异系数为24%，组胺致敏试验的灵敏度为3.3 IU·mL^-1；所有样品的体外定性结果均为阴性，小鼠组胺致敏试验结果均合格，体外定量结果与小鼠组胺致敏结果具有良好的相关性（r=0.737，P＜0.05）。结论：本研究在国内首次使用 CHO细胞簇集试验方法检测百日咳疫苗成品，并进行了初步评价。该方法灵敏度高于小鼠组胺致敏试验，可应用于百日咳疫苗毒性及毒性逆转检测。该方法需要更广泛的推广应用和数据积累，以达到完全替代动物实验的目标。     

PIC/S GMP先进治疗产品附录与我国细胞治疗产品生产质量管理指南的对比分析

目的：为我国申请加入药品检查合作计划(PIC/S),建立与国际统一的检查标准,提出细胞治疗产品GMP检查亟需关注的差异点。方法：对我国细胞治疗产品指南与PIC/S先进治疗产品GMP附录进行整体梳理和对比分析,并就一些关键问题进行了讨论。结果：我国与PIC/S GMP附录原则及考虑要素基本是一致的,但从法规体系、框架结构、适用范围、具体要求均有所区别。结论：分别从质量风险管理、起始生物材料的检查、与国际检查标准的统一三个方面提出了对此类产品GMP检查的启示。     

生物制品早期研发阶段常见的研发质量管理问题及对策

目的：为规范生物制品早期研发工作,提高早期研发质量管理水平提供建议,提升生物制品的研发质量与效率。方法：汇总我国现行与生物制品研发质量管理相关的法规,分析法规要求与生物制品研发现状的契合度,根据生物制品早期研发阶段的特点,概括早期研发过程中存在的研发质量管理问题,提出解决问题的对策与建议。结果：现阶段我国已颁布的与生物制品研发质量管理相关的法规主要涉及研发中后期,对于早期阶段的质量管理,缺少明确、细致、可执行的法规文件。生物制品早期研发过程中在组织架构/人员设置、职能划分、管理制度、制度执行与培训、研究资料、物料和供应商管理以及样品管理7个方面存在问题。结论：加强生物制品早期研发过程的研发质量管理应注意以下几点内容：（1）建立权责明确的生物制品研发质量管理部门;（2）建立层次分明的生物制品研发质量管理文件体系;（3）采取“纵横交错、贴合实际”的原则来制定研发质量管理文件;（4）加强质量管理工具的应用;（5）采用项目管理模式开展生物制品早期研发质量管理;（6）建立与健全研发项目代码编号管理制度,加强早期研发阶段的样品管理;（7）建立与完善研发培训体系。     

预防传染病mRNA疫苗原材料、原液质控要点分析

目的：梳理归纳预防传染病mRNA疫苗原材料和原液质控要点,为我国此类产品质量控制提供参考。方法：通过梳理世界卫生组织及相关标准和审评机构关于预防传染病mRNA疫苗原材料、原液质控指导文件,以及相关监管指南和文献,总结归纳预防传染病mRNA疫苗原材料和原液关键质量控制属性及相关要求。结果与结论：作为新型生物制品,mRNA疫苗核酸结构和脂质体包裹制剂的特性决定了该类疫苗质控特点,序列和完整性、含量及纯度、加帽率和包封率是mRNA疫苗特有的、决定有效性和安全性的关键质量参数。从原材料和原液制备阶段就建立多个与mRNA特性相关的质控要点及其检测方法,将有助于保障预防传染病mRNA疫苗安全有效、质量可控。     

Prediction of lipid nanoparticles for mRNA vaccines by the machine learning algorithm

Lipid nanoparticle (LNP) is commonly used to deliver mRNA vaccines. Currently, LNP optimization primarily relies on screening ionizable lipids by traditional experiments which consumes intensive cost and time. Current study attempts to apply computational methods to accelerate the LNP development for mRNA vaccines. Firstly, 325 data samples of mRNA vaccine LNP formulations with IgG titer were collected. The machine learning algorithm, lightGBM, was used to build a prediction model with good performance (R² > 0.87). More importantly, the critical substructures of ionizable lipids in LNPs were identified by the algorithm, which well agreed with published results. The animal experimental results showed that LNP using DLin-MC3-DMA (MC3) as ionizable lipid with an N/P ratio at 6:1 induced higher efficiency in mice than LNP with SM-102, which was consistent with the model prediction. Molecular dynamic modeling further investigated the molecular mechanism of LNPs used in the experiment. The result showed that the lipid molecules aggregated to form LNPs, and mRNA molecules twined around the LNPs. In summary, the machine learning predictive model for LNP-based mRNA vaccines was first developed, validated by experiments, and further integrated with molecular modeling. The prediction model can be used for virtual screening of LNP formulations in the future.     

Challenges of general safety evaluations of biologics compared to small molecule pharmaceuticals in animal models

Importance of the field: The prediction of human toxicity by employing animal models for nonclinical safety evaluation of pharmaceuticals poses numerous challenges. Each type, biologics, vaccines and small molecules, has unique features, which may impact the ability to effectively assess safety.

Areas covered in this review: The importance of taking a case-by-case approach is highlighted in this review of the challenges encountered in general safety evaluations for biologics and vaccines compared to small molecules.

What the reader will gain: The reader will gain insights in specific issues related to building a successful predictive nonclinical safety program for biologics.

Take home message: While there is fair concordance for small molecules, animal models used for the safety evaluation of biologics may have limitations with regard to human relevance. For small molecules, this is commonly because of differences in metabolism profiles or off-target effects. For biologics, which are highly targeted molecules, it may be because of differences in physiological processes or biologic pathways that limit pharmacologic relevance. For vaccines or immunomodulatory biologics, it may be related to the complexities of modeling the human immune system in a nonhuman species. While international guidances are available to govern the nonclinical safety assessment process for human pharmaceuticals (such as ICH M3), in many instances a case-by-case approach is employed for novel agents.     

Considerations for the nonclinical safety evaluation of antibody drug conjugates for oncology

Antibody drug conjugates (ADCs) include monoclonal antibodies that are linked to cytotoxic small molecules. A number of these agents are currently being developed as anti-cancer agents designed to improve the therapeutic index of the cytotoxin (i.e., cytotoxic small molecule or cytotoxic agent) by specifically delivering it to tumor cells. This paper presents primary considerations for the nonclinical safety evaluation of ADCs and includes strategies for the evaluation of the entire ADC or the various individual components (i.e., antibody, linker or the cytotoxin). Considerations are presented on how to design a nonclinical safety assessment program to identify the on- and off-target toxicities to enable first-in-human (FIH) studies. Specific discussions are also included that provide details as to the need and how to conduct the studies for evaluating ADCs in genetic toxicology, tissue cross-reactivity, safety pharmacology, carcinogenicity, developmental and reproductive toxicology, biotransformation, toxicokinetic monitoring, bioanalytical assays, immunogenicity testing, test article stability and the selection of the FIH dose. Given the complexity of these molecules and our evolving understanding of their properties, there is no single all-encompassing nonclinical strategy. Instead, each ADC should be evaluated on a case-by-case scientifically-based approach that is consistent with ICH and animal research guidelines.     

Lipid-based nanoparticle formulations for small molecules and RNA drugs

ABSTRACT

Introduction: Liposomes and lipid-based nanoparticles (LNPs) effectively deliver cargo molecules to specific tissues, cells, and cellular compartments. Patients benefit from these nanoparticle formulations by altered pharmacokinetic properties, higher efficacy, or reduced side effects. While liposomes are an established delivery option for small molecules, Onpattro® (Sanofi Genzyme, Cambridge, MA) is the first commercially available LNP formulation of a small interfering ribonucleic acid (siRNA).

Areas covered: This review article summarizes key features of liposomal formulations for small molecule drugs and LNP formulations for RNA therapeutics. We describe liposomal formulations that are commercially available or in late-stage clinical development and the most promising LNP formulations for ASOs, siRNAs, saRNA, and mRNA therapeutics.

Expert opinion: Similar to liposomes, LNPs for RNA therapeutics have matured but still possess a niche application status. RNA therapeutics, however, bear an immense hope for difficult to treat diseases and fuel the imagination for further applications of RNA drugs. LNPs face similar challenges as liposomes including limitations in biodistribution, the risk to provoke immune responses, and other toxicities. However, since properties of RNA molecules within the same group are very similar, the entire class of therapeutic molecules would benefit from improvements in a few key parameters of the delivery technology.     

mRNA疫苗的社会价值评估

目的：mRNA疫苗新技术的出现给生物医药企业带来了新的机遇，用定量的方式评价mRNA疫苗的社会价值，以期为其产品的开发提供参考。方法：采用模糊综合评估方法，将mRNA疫苗的社会价值分成正外部性特征价值和负外部性特征影响，将正外部性价值指标分成创新价值、功能性优势、价格优势和其他无形价值，负外部性影响指标则分成对材料的影响、对生产过程的影响、使用或者废弃以后的影响以及对人文环境的影响。同时聘请专家评估社会指标，确定权重并计算社会评价系数。结果：mRNA-1273的社会价值系数为1.84,BNT162b2的社会价值系数为1.98。二者的社会价值系数均远大于1，发展mRNA疫苗总体上利大于弊。结论：mRNA疫苗在生物医药领域具有重要的社会价值，本评估结果可为开发者提供参考，帮助决策者理解其潜在价值，以推动相关技术的进一步发展和应用。     

High dose selection in general toxicity studies for drug development: A pharmaceutical industry perspective

The choice of an appropriate high dose for nonclinical toxicology studies continues to generate significant discussion and debate. Typically, use of the term “high dose” reflects a consideration of a Maximum Tolerated Dose (MTD) or a Maximum Feasible Dose (MFD), inexact terms applied to the design of nonclinical studies conducted to support human clinical trials for experimental new drugs. A pharmaceutical industry perspective on appropriate considerations for high doses in nonclinical studies is provided herein, however, the basic principles applied to the design of toxicology studies translate across the areas of Regulatory, Academic, and Industrial toxicology. Dose selection approaches for nonclinical studies of safety assessment for pharmaceuticals should consider the need to demonstrate the full range of the dose–response continuum (e.g., NOAEL through a toxic dose), however, should also take into account relevance to human therapeutic doses and incorporate clinical indication- and phase-specific considerations.     

Nucleic acid vaccines: prospects for non-viral delivery of mRNA vaccines

Introduction: Nucleic acid-based vaccines are being developed as a means to combine the positive attributes of both live-attenuated and subunit vaccines. Viral vectors and plasmid DNA vaccines have been extensively evaluated in human clinical trials and have been shown to be safe and immunogenic, although none have been licensed for human use. More recently, mRNA-based vaccine alternatives have emerged and might offer certain advantages over their DNA-based counterparts.

Areas covered: This review describes the two main categories of mRNA vaccines: conventional non-amplifying and self-amplifying mRNA. It summarizes the initial clinical proof-of-concept studies and outlines the preclinical testing of the next wave of innovations for the technology. Finally, this review highlights the versatile functionality of the mRNA molecule and introduces opportunities for future improvements in vaccine design.

Expert opinion: The prospects for mRNA vaccines are very promising. Like other types of nucleic acid vaccines, mRNA vaccines have the potential to combine the positive attributes of live attenuated vaccines while obviating many potential safety limitations. Although data from initial clinical trials appear encouraging, mRNA vaccines are far from a commercial product. These initial approaches have spurred innovations in vector design, non-viral delivery, large-scale production and purification of mRNA to quickly move the technology forward. Some improvements have already been tested in preclinical models for both prophylactic and therapeutic vaccine targets and have demonstrated their ability to elicit potent and broad immune responses, including functional antibodies, type 1 T helper cells-type T cell responses and cytotoxic T cells. Though the initial barriers for this nucleic acid vaccine approach seem to be overcome, in our opinion, the future and continued success of this approach lies in a more extensive evaluation of the many non-viral delivery systems described in the literature and gaining a better understanding of the mechanism of action to allow rational design of next generation technologies.     

超微结构病理学在药物非临床安全性评价毒理学研究中的应用现状和关注点

超微结构病理学是药物非临床毒理学研究中毒性病理学评估的重要辅助工具。简要介绍了超微结构病理学在药物非临床安全性评价毒理学研究中的应用现状和关注点,并举例说明了透射电子显微镜等超微结构病理学技术在药物非临床安全性评价中的应用。目前,超微结构病理学技术在药物非临床毒理研究中虽然使用率不高,却是非临床毒理学研究中毒性病理学评估的重要辅助工具,可用于对光学显微镜检查结果的进一步研究,以作为早期药物发现和非临床安全性评价毒理学研究的有益补充。在使用超微结构病理学技术进行药物非临床毒理学研究时,应关注其使用的必要性、良好实验室规范（GLP）依从性、专业性、科学性和使用局限性等方面。