肿瘤新生抗原多肽具有高度的免疫原性和肿瘤特异性,是肿瘤精准化治疗的理想策略之一. 然而,受其个性化的特征限制,目前临床研究肿瘤新生抗原多肽疫苗的生产中面临合成难度大、制备周期长和纯化复杂等挑战,限制了其广泛应用. 针对目前面临的问题,本研究提出了一种全新的利用片段肽树脂储存的方式,将合成周期由2个月缩短至1个月;并对合成方法进行优化,大幅提高粗肽纯度,降低纯化难度. 利用该方法,可以在23个工作日内高效生产12条38个氨基酸长度的肿瘤新生抗原多肽,以满足临床需求（纯度大于95%,质量大于20 mg,转成醋酸盐且三氟乙酸根残留小于1%,内毒素残留小于10 EU/mg）. 本研究在新生抗原多肽制备流程的设计与优化方面取得了显著进展,不仅提高了合成肿瘤新生抗原多肽的质量,还显著缩短了制备周期,为肿瘤新生抗原多肽临床应用奠定了坚实基础.
Liu
,
Qiang
,
Liu
,
Xinyao
,
Zu
,
Yuan
,
Hu
,
Hongguo
,
Yao
,
Wenbing
. 肿瘤新生抗原多肽疫苗制备方法研究[J]. 化学学报, 0
: 250613
.
DOI: 10.6023/A25040126
Tumor neoantigen peptides, characterized by tumor-specific mutations and aberrant gene expression profiles, demonstrate superior immunogenicity and minimal off-target effects. Neoantigen peptide vaccines are able to activate the immune system, break immune tolerance, and induce tumor cell cytotoxicity through enhanced antigen-specific T-cell responses, thereby emerging as promising candidates for personalized cancer immunotherapy. However, due to their personalized characteristics, the current commercial production of tumor neoantigen peptide vaccines face challenges in difficult synthesis, long preparation cycles and complex purification processes, which limit their widespread clinical applications. To address these issues, this study proposes a novel method utilizing fragmented peptide resin storage, in which the shared Link-NitraTh peptide sequence is first synthesized, and then it is used as the starting material for the subsequent preparation of full-length neoantigen peptides. This approach reduces the overall synthesis cycle from two months to one month, significantly shortening the production time for synthetic peptides and increasing the accessibility of neoantigen peptide vaccines for clinical patients; Additionally, optimization of the shared Link-NitraTh synthesis method greatly improves the purity of crude neoantigen peptides and simplifies the purification process, enabling robust production of neoantigen peptide vaccines with stringent quality control and acceptable manufacturing time. Using this method, twelve tumor neoantigen peptides, each comprising thirty-eight amino acids, can be efficiently produced within twenty-three working days to meet clinical demands. The quality specifications for neoantigen peptides include: purity >95% by PHLC, conversion to the acetate salt form and trifluoroacetate residue <1%, mass exceeding 20 mg, and endotoxin level below 10 EU/mg. This work has achieved significant progress in the design and optimization of the tumor neoantigen peptide preparation process, and established a standardized operational framework including parameter optimization for peptide solid-phase synthesis and liquid-phase purification protocols. Our optimized methodologies not only improve the synthesis efficiency and enhance batch-to-batch consistency of tumor neoantigen peptides, but also reduce production timelines. These advancements represent a pivotal step toward realizing the therapeutic potential of patient-specific neoantigen peptide vaccine platforms and accelerating their clinical translation within precision medicine frameworks for oncological and immunological indications.
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