Oosting Linette T, Franke Katka, Martin Michael V, Kloosterman Wigard P, Jamieson Jennifer A, Glenn Laura A, de Jager Miranda W, van Zanten Jacoba, Allersma Derk P, Gareb Bahez
Department of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
CureVac Netherlands B.V., Matrix Building VII, Science Park 106, 1098 XG Amsterdam, The Netherlands.
Pharmaceutics. 2022 Jul 21;14(7):1515. doi: 10.3390/pharmaceutics14071515.
Stage III-IV non-small cell lung cancer (NSCLC) is a devastating disease characterized by a poor prognosis. NSCLC tumors carry genetic mutations, which can lead to the expression of altered protein sequences. Peptides originating from mutated proteins and bound to MHC molecules on the tumor cell surface are referred to as neoantigens, as they are tumor-specific and not expressed in normal cells. Due to their tumor specificity, neoantigens have a strong potential to induce an anti-tumor immune response and have been investigated for development of personalized therapeutic cancer vaccines. The current study describes the development of a clinical grade neoantigen vaccine formulation (FRAME-001) intended as immunotherapy in advanced NSCLC in combination with the immune checkpoint inhibitor pembrolizumab. The detection of aberrant tumor-specific transcripts as well as an algorithm to select immunogenic neoantigen peptides are described. Subsequently, selected neoantigen peptides were synthesized with a high throughput synthesis platform and aseptically formulated under good manufacturing practice (GMP) conditions into four aqueous peptides mixtures that each contained six neoantigen peptides. A validated stability-indicating analytical method was developed in which we considered the personalized nature of the formulation. An extensive stability study performed either at -25 °C or -80 °C showed that the formulation was stable for up to 32 weeks. The formulation was mixed with the vaccine adjuvant Montanide ISA 51 VG, which yielded the final vaccine emulsion. The stability of the vaccine emulsion was demonstrated using microscopic examination, differential light scattering, and the water-drop test. The presented data show that FRAME-001 is a feasible personalized vaccine formulation for the treatment of stage III-IV NSCLC. The presented data may give guidance in the development of novel personalized therapeutic vaccines since this formulation strategy could be used for any cancer indication.
III-IV期非小细胞肺癌（NSCLC）是一种预后较差的毁灭性疾病。NSCLC肿瘤携带基因突变，这可能导致蛋白质序列改变的表达。源自突变蛋白并与肿瘤细胞表面的MHC分子结合的肽被称为新抗原，因为它们是肿瘤特异性的，在正常细胞中不表达。由于其肿瘤特异性，新抗原有很强的诱导抗肿瘤免疫反应的潜力，并已被研究用于开发个性化治疗性癌症疫苗。本研究描述了一种临床级新抗原疫苗制剂（FRAME-001）的开发，该制剂旨在与免疫检查点抑制剂帕博利珠单抗联合用于晚期NSCLC的免疫治疗。文中描述了异常肿瘤特异性转录本的检测以及选择免疫原性新抗原肽的算法。随后，使用高通量合成平台合成选定的新抗原肽，并在良好生产规范（GMP）条件下无菌配制为四种水性肽混合物，每种混合物包含六种新抗原肽。我们开发了一种经过验证的稳定性指示分析方法，其中考虑了制剂的个性化性质。在-25°C或-80°C下进行的广泛稳定性研究表明，该制剂在长达32周内稳定。该制剂与疫苗佐剂Montanide ISA 51 VG混合，制成最终的疫苗乳剂。通过显微镜检查、差示光散射和水滴试验证明了疫苗乳剂的稳定性。所呈现的数据表明，FRAME-001是一种用于治疗III-IV期NSCLC的可行的个性化疫苗制剂。所呈现的数据可能为新型个性化治疗性疫苗的开发提供指导，因为这种制剂策略可用于任何癌症适应症。
