作者:巫嘉美 薛超然美工:何国红 罗真真排版:马超01 引言多肽药物以其特异性高、免疫原性低、毒副作用小等显著优势,在药物研发领域展现出巨大潜力,可有效作用于胞内、胞外、膜表面等多样化靶点。三优生物凭借对天然多肽序列的深度分析,融合具有自主知识产权的标签蛋白,并辅以尖端AI设计,运用噬菌体展示技术,结合真核表达与高通量筛选,隆重推出超万亿多肽分子发现平台,致力于为全球医药研发提供高效、前沿的环状多肽分子产生系统解决方案。02 文库背景根据弗若斯特沙利文(Frost & Sullivan)等机构的数据,全球多肽药物市场近年来持续增长:2020年全球多肽药物市场规模达到628亿美元。预计到2025年,全球多肽药物市场规模将增至960亿美元。长期来看,预计到2030年,全球多肽药物市场规模可能达到1418亿美元甚至2108亿美元(不同来源预测略有差异,但都显示出强劲的增长趋势)。这些数据表明,多肽药物市场具有巨大的增长空间。中国多肽药物市场也呈现逐年递增趋势,增速远超全球平均水平。多肽类药物之所以备受瞩目,在于其显著优势:它们不仅特异性高、免疫原性较低,而且毒副作用小,尤其适用于胞内靶点,为诸多疾病治疗提供了新的可能。然而,多肽药物研发的核心挑战之一在于构建高通量、多样化的多肽文库。对于大多数药物研发公司而言,这一过程门槛高,花费大,周期长。为攻克这一瓶颈,三优生物凭借前瞻性布局和持续性研发投入,于2022年成功构建并验证了千亿级多肽文库,并在此基础上,于2023年进一步完成了万亿级多肽文库的构建与验证。这一里程碑式的突破,标志着三优生物在多肽药物发现领域迈入了全新阶段,为全球医药研发提供了高效、前沿的环状多肽分子产生系统解决方案。03 文库组成三优生物倾力打造的环状多肽库,深度遵循自然规律,辅以尖端AI设计,巧妙融合独有标签蛋白与噬菌体展示技术,并匹配全面的理化及生化分析验证,构建出包含17个子库、多肽长度覆盖6-17个氨基酸、总库容高达3.05×1012的超万亿级分子发现平台。该平台特别优选标签蛋白与天然环肽融合,通过筛选可以获得中位数100+的序列独特先导分子。不仅适用于代谢、肿瘤等疾病靶点的新型多肽药物研发,亦可广泛应用于医学影像、分子诊断等前沿领域,并可进一步形成PDC、RDC、抗体融合蛋白、多环肽等多种新型药物。▼ 表1. 万亿级环状多肽库相关统计04 文库特性01序列遵循自然规律,多样性高三优生物的多肽库设计,融合了自然规律、人工智能(AI)和前沿生物技术。收集并深入分析海量天然多肽序列,从中提取氨基酸分布规律。在此基础上,通过AI算法进行理性设计与优化,极大地拓展了多肽分子的多样性和成药性潜力。图1和图2显示三优生物NL17和NB17肽氨基酸分布频率,符合自然规律,序列多样性高。▲ 图1. NL17 AA 频率▲ 图2. NB17 AA 频率02融合自主知识产权的标签,高效表达纯化三优生物凭借自主知识产权的特殊融合标签蛋白,有效解决了多肽类分子的高效表达与纯化难题。该标签不仅能显著提升多肽在宿主系统中的可溶性表达量和展示效率,还通过其特异性结合能力,实现了下游批量纯化,极大简化了多肽药物研发的复杂流程,加速了高品质先导分子的获取。▲ 图3. 多肽-融合蛋白噬菌体展示示意图03噬菌体展示效率高噬菌体展示技术是三优生物高效筛选多肽库的关键。通过对上千个多肽库克隆进行严格检测,结果如图4所示,证实其噬菌体展示效率高达98%。高展示效率不仅能够减少文库中的无效分子的占比,提高文库的品质,也在一定程度上表明了所设计多肽可以高效表达,成药性好。▲ 图4. 多肽库展示效率04二硫键成环,结构刚性稳定三优生物环状多肽库采用关键的二硫键成环策略,相较于线性多肽,二硫键形成的环状结构能显著限制构象自由度,大幅提升多肽的结构刚性和稳定性,从而增强其对蛋白酶降解的抵抗力,延长体内半衰期并提高生物利用度。与其他化学成环方式(如酰胺键、醚键等)相比,二硫键成环不仅因其在天然蛋白质中的广泛存在而具有优异的生物兼容性,更可精确锁定多肽的活性构象,赋予其更高的靶点亲和力与选择性,为药物发现提供更具潜力的分子骨架。筛选多肽进行半胱氨酸突变,结果如图5所示,突变前多肽成环,有结合活性;突变后环肽线性化,无结合活性。▲ 图5. 多肽环化验证05多样筛选策略匹配,高通量筛选平台三优生物采用一站式高通量自动化筛选,极大的缩短了筛选时间,仅需6-8周即可获得特异性好、亲和力高、序列明确并经真核表达验证的多肽,针对不同抗原可制定Fc/His标签交叉、固液交叉、细胞蛋白交叉竞争及阻断筛选等多种筛选策略,采用固、液相高通量快速获得先导分子。将先导分子进行穿梭构建,通过CHO/293真核表达系统进行表达,进行生化理化分析以及细胞水平的功能验证,获得真实有效的成药性数据。▲ 图6. 多肽筛选流程05 代表案例01PD-L1靶点背景基本信息:PD-L1,具有免疫球蛋白结构的I型单次跨膜糖蛋白。PD-L1与PD-1结合后传递抑制信号,减少淋巴结中抗原特异性T细胞的增殖和调节性T细胞的凋亡。PD-L1在多种实体瘤细胞表面高表达。竞争格局:全球以PD-L1为靶点的抗体药物已有多款上市,国外基因泰克的阿替利珠单抗,默克的阿维单抗,国内基石药业的舒格利单抗,康宁杰瑞的恩沃利单抗。药物MOA:靶向PD-L1的抗体能够抑制PD-L1与PD-1的结合,激活T细胞对肿瘤的杀伤效应,抑制PD-L1和PD-1的结合可能导致T细胞凋亡、无能、衰竭和IL-10表达。02抗PD-L1抗体关键结果◆ 亲和活性分析将万亿多肽库筛选到的多肽分子,经过亲和力成熟后,用huPD-L1抗原蛋白进行ELISA亲和活性分析,结果如图所示,多肽库筛选到的多肽分子特异结合huPD-L1抗原,经过亲和力成熟后,抗原结合EC50约为0.04 nM,与对照抗体亲和力相当。在huPD-L1-CHO-K细胞上进行FACS亲和活性分析,结果如图所示,万亿多肽库筛选到的多肽分子,经过亲和力成熟后,结合huPD-L1-CHO-K细胞的EC50为1-10 nM。▲ 图7. 多肽分子ELISA亲和活性检测▲ 图8. 多肽分子FACS亲和活性检测◆ 阻断活性分析通过万亿多肽库筛选到的多肽分子,经过亲和力成熟后,在huPD-L1-CHO-K细胞上进行FACS亲和活性分析,结果如图所示,万亿多肽库筛选到的多肽分子,经过亲和力成熟后,阻断huPD1与huPD-L1-CHO-K细胞的EC50为5.7 nM。▲ 图9. 多肽分子FACS阻断活性06 总结展望当前,全球多肽药物市场正经历爆发式增长,仅2023年司美格鲁肽的全球销售额便接近200亿美元,预示着该领域巨大的增长空间和未被满足的临床需求。多肽文库的建立是多肽药物研发的核心,亦是公认的技术高点。三优生物凭借在创新生物药领域多年的深厚积累,并依托成功搭建的STAL超万亿抗体库平台的经验,已攻克多肽库构建的各项技术瓶颈,成功完成了超万亿多肽分子发现平台的构建和验证。展望未来,三优生物将持续发挥其平台优势,致力于攻克多肽药物研发中的关键难点,包括引入非天然氨基酸以提升多肽的口服生物利用度,开发高效穿膜多肽以靶向胞内靶点,以及运用从头设计(de novo design)策略优化环状多肽的成药性。通过这些创新举措,三优生物将显著降低多肽药物的开发难度,极大加速创新多肽类药物的研发进程,为全球患者带来更多高效、安全的治疗选择。Sanyou Bio AI-Powered Super Trillion Molecule Discovery Platform: Cyclic Peptide Molecule Discovery Solution (One of Sanyou’s Six Molecule Libraries)01 IntroductionPeptide therapeutics demonstrate significant potential in drug development due to their high specificity, low immunogenicity, and favorable safety profiles. These molecules effectively engage diverse targets, including intracellular, extracellular, and membrane-bound proteins. Sanyou Bio just launched its trillion-scale peptide discovery platform, which integrates deep analysis of natural peptide sequences,proprietary fusion tag technology,AI-driven molecular design, phage display coupled with eukaryotic expression and high-throughput screening. This cutting-edge solution delivers efficient cyclic peptides for global pharmaceutical R&D.02 Library BackgroundGlobal peptide therapeutics market continues robust growth, with revenues reaching $62.8 billion in 2020 (Frost & Sullivan data). Projections indicate expansion to $96 billion by 2025, potentially reaching $141.8-$210.8 billion by 2030, reflecting strong upward trajectories across forecasts.The sector's prominence stems from peptide advantages: high target specificity,low immunogenicity,favorable safety profiles as well as particular effectiveness against intracellular targets.Nevertheless,core industry challenge is that the development of diverse and high-throughput peptide libraries has been high-cost and time-intensive. Sanyou Bio validated 100-billion-scale peptide library in 2022 and further achieved trillion-scale library construction & validation in 2023. This milestone establishes an efficient cyclic peptide solution for global drug discovery.03 Library CompositionSanyou Bio's trillion-scale cyclic peptide discovery platform integrates natural peptide principles with AI-driven design, combining proprietary fusion tags and phage display technology validated through comprehensive physicochemical and biochemical analyses. This engineered system comprises 17 sublibraries spanning 6-17 amino acid lengths, delivering a total diversity of 3.05 trillion molecules.This platform employs specifically engineered fusion tags for natural cyclic peptides, enabling the identification of lead molecules with a median yield of >100 structurally distinct candidates per screen. These molecules are applicable to novel peptide drug development for metabolic disorders, oncology, and other therapeutic targets, as well as medical imaging and molecular diagnostics, with further potential for generating PDC (peptide-drug conjugates), RDC (radiopharmaceutical conjugates), antibody fusion proteins, and multicyclic peptides as advanced therapeutic modalities.▼ Table 1. Statistics related to trillion level cyclic peptide libraries04 Library Characteristics01Natural sequence principles with high diversitySanyou Bio's peptide library design integrates natural principles, artificial intelligence, and cutting-edge biotechnology. By analyzing extensive natural peptide sequences to extract amino acid distribution patterns, followed by AI-driven rational design and optimization, the platform significantly enhances molecular diversity and drug developability. Figures 1-2 demonstrate that NL17/NB17 peptide amino acid frequencies align with natural distribution while maintaining high sequence diversity.▲ Fig 1. NL17 AA frequency▲ Fig 2. NB17 AA frequency02Proprietary fusion tags enabling high-yield expression and streamlined purificationBy leveraging proprietary fusion tag technology, Sanyou Bio effectively addresses critical challenges in peptide expression and purification. This engineered tag significantly enhances soluble expression yield and display efficiency in host systems while enabling batch purification based on affinity, thereby streamlining peptide drug development.▲ Fig 3. Schematic diagram of peptide fusion protein phage display03High-efficiency phage displayOptimized phage display technology underpins Sanyou Bio's efficient peptide library screening. Thorough evaluation of thousands of clones (Fig. 4) validates >98% display efficiency, which minimizes the presence of non-functional molecules and enhances library quality.▲ Fig 4. Peptide library display efficiency04Disulfide cyclization for structural rigidity and stabilityThe cyclic peptide library employs disulfide bond cyclization to significantly restrict conformational freedom compared to linear peptides. This structural rigidity enhances protease resistance, prolongs in vivo half-life, and improves bioavailability through stabilized tertiary structures.Disulfide cyclization outperforms non-disulfide methods (e.g., amide/ether bonds) with superior biocompatibility derived from natural protein prevalence. It precisely locks bioactive conformations, conferring higher target affinity/selectivity for discovery-phase scaffolds. Cysteine mutation studies (Fig. 5) confirm functional loss upon linearization.▲ Fig 5. Peptide cyclization validation05Multiplexed screening strategies coupled with high-throughput platformSanyou Bio integrated high-throughput platform to accelerate screening to 6-8 weeks, delivering high-specificity, high-affinity peptides with validated sequences. Customizable strategies include: Fc/His tag cross-screening, solid/liquid-phase cross-screening, cell/protein cross-screening and cellular blocking functional screening. Lead molecules undergo CHO/293 expression with biochemical and cell-based functional validation for robust developability data.▲ Fig 6. Peptide screening process05 Representative Case: PD-L101Target BackgroundProgrammed Death-Ligand 1 (PD-L1) is a Type I single-pass transmembrane glycoprotein with immunoglobulin superfamily structure. Its binding to PD-1 delivers inhibitory signals that reduce antigen-specific T-cell proliferation in lymph nodes and promote regulatory T-cell apoptosis. PD-L1 is overexpressed on multiple solid tumor surfaces.Competitive Landscape: Globally approved PD-L1 antibodies include Roche/Genentech's atezolizumab and Merck's avelumab. In China, CStone Pharmaceuticals' sugemalimab and Alphamab Oncology's envafolimab (the first subcutaneous PD-L1 inhibitor) have gained market approval.Therapeutic Mechanism: PD-L1 antibodies block PD-1/PD-L1 interactions, reactivating T-cell-mediated tumor killing. Incomplete blockade may induce T-cell apoptosis, anergy, exhaustion, and IL-10 overexpression.02Key Results of Anti-PD-L1 Peptide Program◆ Affinity AnalysisLead peptide candidates from the trillion-scale library demonstrated specific binding to huPD-L1 in ELISA. Post-affinity maturation, the EC50 reached 0.04 nM – comparable to reference antibodies. FACS binding assays on huPD-L1-CHO-K cells confirmed EC50 values of 1-10 nM.▲ Fig 7. Peptide molecule ELISA affinity activity detection▲ Fig 8. Peptide molecule FACS affinity activity detection◆ Blocking ActivityFACS-based blocking assays revealed an EC50 of 5.7 nM for inhibiting PD-1/PD-L1 interactions on huPD-L1-CHO-K cells after affinity maturation.▲ Fig 9. Peptide molecule FACS blocking activity06 Outlook and PerspectivesThe global peptide therapeutics market is experiencing exponential growth, exemplified by semaglutide (Ozempic®) achieving nearly $20 billion in 2023 sales, highlighting substantial expansion opportunities and unmet clinical needs. Peptide library construction remains the cornerstone of drug discovery and a critical technological frontier. Leveraging expertise from its STAL trillion-scale antibody platform, Sanyou Bio has overcome key technical hurdles to establish and validate an ultra-trillion peptide discovery system.Looking forward, Sanyou Bio will harness this platform to address core challenges: enhancing oral bioavailability through non-canonical amino acid incorporation,developing cell-penetrating peptides for intracellular targets, optimizing developability via de novo cyclic peptide design. These innovations will streamline peptide drug development pipelines and accelerate novel therapeutics delivery to patients worldwide.推荐阅读轻松玩转三优oneClick+线上9大云程序三优6类分子形式之mRNA分子产生系统解决方案三重好礼相送暨四种抗体制备解决方案全新上线三优生物单域抗体产生系统解决方案三优生物双抗参比品网站SY-BsAb正式上线三优ADC药物研发系统解决方案三优生物oneClick+平台再次上新三优磁阵列全人源小鼠抗体发现平台重磅发布三优超万亿全人单克隆抗体产生平台盘点三优磁阵列全人源小鼠抗体发现平台隆重上线共同轻链抗体产生之超万亿共轻库盘点三优生物73种全系列双抗参比品全新上线三优生物智能超万亿分子发现平台盘点及展望十年磨一剑之三优智能百万亿分子库发展历程三优生物2024|夯基筑本AI-STAL 1.0关于三优生物三优生物是一家以“让天下没有难做的创新生物药”为使命,以超万亿分子库和人工智能技术双驱的生物医药智能高新技术企业。公司以智能超万亿分子库为核心,打造了干湿结合、国际领先的创新生物药临床前智能化及一体化研发平台,通过“新药发现、临床前研究、智能化药物研发及前沿科学研究”等四个维度加速全球新药发现及靶标深度研究。公司总部位于中国上海,在美国、欧洲等地设有子公司,现有投产及布局的研发及GMP场地20000多平方米。公司为合作伙伴提供“差异化CRO、整合型CDO、协同型CPO、特色CRS”于一体的“创新生物药4C综合业务”。公司已建立全球营销网络,已与全球1200多家药企、生技公司等建立了良好的合作关系;已完成了1200多个新药发现及开发服务项目;已完成了50多个合作研发项目,其中9个合作项目已完成临床申报。公司已获得国家高新技术、上海市专精特新、上海市小巨人和上海“张江之星”企业认定。