Integrating molecular dynamics simulation with small‐ and wide‐angle X‐ray scattering to unravel the flexibility, antigen‐blocking, and protease‐restoring functions in a hindrance‐based pro‐antibody

Article

作者: Jeng, U‐Ser ; Liao, Jun Min ; Ho, Kai‐Wen ; Shih, Orion ; Hu, I‐Chen ; Lyu, Ping‐Chiang ; Chang, Chin‐Yuan ; Toh, Shu‐Ing ; Cheng, Tian‐Lu ; Wang, Yeng‐Tseng ; Huang, Ming‐Yii ; Hong, Shih‐Ting ; Cheng, Yi‐An

AbstractSpatial hindrance‐based pro‐antibodies (pro‐Abs) are engineered antibodies to reduce monoclonal antibodies' (mAbs) on‐target toxicity using universal designed blocking segments that mask mAb antigen‐binding sites through spatial hindrance. By linking through protease substrates and linkers, these blocking segments can be removed site‐specifically. Although many types of blocking segments have been developed, such as coiled‐coil and hinge‐based Ab locks, the molecular structure of the pro‐Ab, particularly the region showing how the blocking fragment blocks the mAb, has not been elucidated by X‐ray crystallography or cryo‐EM. To achieve maximal effect, a pro‐Ab must have high antigen‐blocking and protease‐restoring efficiencies, but the unclear structure limits its further optimization. Here, we utilized molecular dynamics (MD) simulations to study the dynamic structures of a hinge‐based Ab lock pro‐Ab, pro‐Nivolumab, and validated the simulated structures with small‐ and wide‐angle X‐ray scattering (SWAXS). The MD results were closely consistent with SWAXS data (χ2best‐fit = 1.845, χ2allMD = 3.080). The further analysis shows a pronounced flexibility of the Ab lock (root‐mean‐square deviation = 10.90 Å), yet it still masks the important antigen‐binding residues by 57.3%–88.4%, explaining its 250‐folded antigen‐blocking efficiency. The introduced protease accessible surface area method affirmed better protease efficiency for light chain (33.03 Å2) over heavy chain (5.06 Å2), which aligns with the experiments. Overall, we developed MD‐SWAXS validation method to study the dynamics of flexible blocking segments and introduced methodologies to estimate their antigen‐blocking and protease‐restoring efficiencies, which would potentially be advancing the clinical applications of any spatial hindrance‐based pro‐Ab.

2024-08-01·Inflammatory Bowel Diseases

Monoclonal Antibody Against Mature Interleukin-18 Ameliorates Colitis in Mice and Improves Epithelial Barrier Function

Article

作者: Sawada, Tsunaki ; Urano, Takeshi ; Muto, Hisanori ; Ohno, Eizaburo ; Ishikawa, Eri ; Furukawa, Kazuhiro ; Maeda, Keiko ; Iida, Tadashi ; Yamamoto, Kenta ; Oishi, Akina ; Mizutani, Yasuyuki ; Ikegami, Shuji ; Ishikawa, Takuya ; Honda, Takashi ; Mu, Jingxi ; Yamamura, Takeshi ; Kakushima, Naomi ; Nakamura, Masanao ; Kawashima, Hiroki ; Ishigami, Masatoshi

AbstractBackgroundAntitumor necrosis factor (TNF)-α antibodies have improved the outcome of inflammatory bowel disease (IBD); but half of patients remain unresponsive to treatment. Interleukin-18 (IL-18) gene polymorphism is associated with resistance to anti-TNF-α antibodies, but therapies targeting IL-18 have not been clinically applied. Only the mature protein is biologically active, and we aimed to investigate whether specific inhibition of mature IL-18 using a monoclonal antibody (mAb) against a neoepitope of caspase-cleaved mature IL-18 could be an innovative treatment for IBD.MethodsThe expression of precursor and mature IL-18 in patients with UC was examined. Colitis was induced in C57/BL6 mice by administering dextran sulfate sodium (DSS), followed by injection with anti-IL-18 neoepitope mAb. Colon tissues were collected and subjected to histological analysis, immunohistochemistry, immunoblotting, and quantitative polymerase chain reaction. Colon epithelial permeability and microbiota composition were analyzed.ResultsMature IL-18 expression was elevated in colon tissues of patients with active ulcerative colitis. Administration of anti-IL-18 neoepitope mAb ameliorated acute and chronic DSS-induced colitis; reduced interferon-γ, TNF-α, and chemokine (CXC motif) ligand-2 production and epithelial cell permeability; promoted goblet cell function; and altered the intestinal microbiome composition. The suppressive effect of anti-IL-18 neoepitope mAb was superior to that of anti-whole IL-18 mAb. Furthermore, combination therapy with anti-TNF-α Ab suppressed acute and chronic colitis additively by suppressing cytokine expressions and reducing cell permeability by upregulating claudin1 and occludin expression.ConclusionsAnti-IL-18 neoepitope mAb ameliorates acute and chronic colitis, suggesting that this mAb will be an innovative therapeutic option for IBD.

2024-03-15·International Journal of Cancer

α‐FAtE: A new predictive score of response to atezolizumab plus bevacizumab for unresectable hepatocellular carcinoma

Article

作者: Himmelsbach, Vera ; Atsukawa, Masanori ; Nishida, Naoshi ; Vitiello, Francesco ; Tajiri, Kazuto ; Fukunishi, Shinya ; Burgio, Valentina ; Aoki, Tomoko ; Niizeki, Takashi ; Itokawa, Norio ; Scartozzi, Mario ; Sakamoto, Naoya ; Hiraoka, Atsushi ; Ochi, Hironori ; Presa, José ; Persano, Mara ; Soldà, Caterina ; Tsuji, Kunihiko ; Tsutsui, Akemi ; Montes, Margarida ; Ohama, Hideko ; Nouso, Kazuhiro ; Yasuda, Satoshi ; Kudo, Masatoshi ; Sho, Takuya ; Kaibori, Masaki ; Koizumi, Yohei ; Ishikawa, Toru ; Tani, Joji ; Cabibbo, Giuseppe ; Hirooka, Masashi ; Iijima, Hiroko ; Imai, Michitaka ; Masi, Gianluca ; Nakamura, Shinichiro ; Tada, Toshifumi ; Itobayashi, Ei ; Casadei-Gardini, Andrea ; Kumada, Takashi ; Foschi, Francesco Giuseppe ; Iwamoto, Hideki ; Iavarone, Massimo ; Naganuma, Atsushi ; Vivaldi, Caterina ; Kariyama, Kazuya ; Okubo, Tomomi ; Hatanaka, Takeshi ; Chon, Hong Jae ; Finkelmeier, Fabian ; Rimini, Margherita ; Suda, Goki ; Morishita, Asahiro ; Kakizaki, Satoru ; Cheon, Jaekyung ; Nishimura, Takashi ; Hiasa, Yoichi ; Lim, Ho Yeong ; Ogawa, Chikara ; Cascinu, Stefano ; Steup, Christoph ; Bergamo, Francesca ; Nagano, Takuya ; Toyoda, Hidenori ; Shimada, Noritomo ; Kosaka, Hisashi ; Amadeo, Elisabeth ; Yoo, Changhoon ; Shimose, Shigeo ; Takaguchi, Koichi ; Kawata, Kazuhito ; Piscaglia, Fabio ; Rossari, Federico ; Tada, Fujimasa

AbstractAtezolizumab plus bevacizumab (AB) and lenvatinib can be alternatively used as first‐line systemic treatment of unresectable hepatocellular carcinoma (HCC). However, no direct comparison of the two regimens has been performed in randomized clinical trials, making the identification of baseline differential predictors of response of major relevance to tailor the best therapeutic option to each patient. Baseline clinical and laboratory characteristics of real‐world AB‐treated HCC patients were analyzed in uni‐ and multivariate analyses to find potential prognostic factors of overall survival (OS). Significant variables were incorporated in a composite score (α‐FAtE) and it was tested for specificity and sensitivity in receiver operating characteristic (ROC) curve and in multivariate analysis for OS. The score was applied in uni‐ and multivariate analyses for OS of a comparable lenvatinib‐treated HCC population. Finally, comparison between treatments was performed in patients with low and high α‐FAtE scores and predictivity estimated by interaction analysis. Time‐to‐progression (TTP) was a secondary endpoint. OS of AB‐treated HCC patients was statistically longer in those with α‐fetoprotein <400 ng/mL (HR 0.62, p = .0407), alkaline phosphatase (ALP) <125 IU/L (HR 0.52, p = .0189) and eosinophil count ≥70/μL (HR 0.46, p = .0013). The α‐FAtE score was generated by the sum of single points attributed to each variable among the above reported. In ROC curve analysis, superior sensitivity and specificity were achieved by the score compared to individual variables (AUC 0.794, p < .02). Patients with high score had longer OS (HR 0.44, p = .0009) and TTP (HR 0.34, p < .0001) compared to low score if treated with AB, but not with lenvatinib. Overall, AB was superior to lenvatinib in high score patients (HR 0.55, p = .0043) and inferior in low score ones (HR 1.75, p = .0227). At interaction test, low α‐FAtE score resulted as negative predictive factor of response to AB (p = .0004). In conclusion, α‐FAtE is a novel prognostic and predictive score of response to first‐line AB for HCC patients that, if validated in prospective studies, could drive therapeutic choice between lenvatinib and AB.

项与 Pro Ab (GlycoNex) 相关的新闻（医药）

2024-08-19

·PRNewswire

GlycoNex Successfully Completes Phase 1 Study of Denosumab Biosimilar, SPD8

GlycoNex plans to initiate Phase 3 clinical program of SPD8 in osteoporosis, targeting a multibillion USD global market opportunity NEW TAIPEI CITY, Taiwan, Aug. 19, 2024 /PRNewswire/ -- GlycoNex (4168, hereinafter referred to as GNX), a clinical stage biotechnology company focused on the development of glycan-directed cancer therapies, today announced the successful completion of a Phase 1 clinical trial of its denosumab biosimilar, SPD8, developed in collaboration with Mitsubishi Gas Chemical Company, Inc. (MGC). The results enable the company to advance to Phase 3 clinical trials with the aim of offering a cost-effective alternative to the currently available denosumab treatment for osteoporosis. The Phase 1 study, conducted in healthy postmenopausal women, demonstrated that GlycoNex's denosumab biosimilar met the primary endpoint of clinical pharmacokinetic equivalence. The trial confirmed the biosimilar's pharmacokinetic profile, safety, and pharmacodynamics were comparable to the reference biologic, denosumab. With these positive results, GlycoNex is in the final stages of planning for its Phase 3 clinical program for SPD8, which is expected to commence in Q4 2024. This program will further assess the efficacy, safety, and immunogenicity of SPD8 in a larger patient population with osteoporosis. GlycoNex is working closely with regulatory authorities to ensure the trial design meets all necessary requirements for a comprehensive evaluation of the biosimilar's clinical performance. "The successful completion of the SPD8 Phase 1 study is an important milestone in the ongoing development of a denosumab biosimilar for the treatment of osteoporosis and GlycoNex's overarching mission to make advanced biologic therapies more accessible and affordable," said Mei-Chun Yang, CEO of GlycoNex. "As we move forward to Phase 3, we remain committed to delivering a safe and effective, high-quality denosumab biosimilar that offers the potential to improve patient outcomes, while reducing the financial burden on osteoporosis patients and healthcare systems around the world." Global sales of two denosumab-containing drugs amounted to 4 billion USD and 2.1 billion USD respectively in 2023. With the increasing elderly population and growing awareness of osteoporosis risks, the market for denosumab continues to expand. In addition to the Japanese market, GlycoNex is actively pursuing global development plans, aiming to provide improved treatment options to people with osteoporosis worldwide. GlycoNex, in collaboration with Mitsubishi Gas Chemical (MGC), is advancing the development and production of SPD8. Recently, MGC and GlycoNex successfully completed the Phase 1 clinical trial of SPD8 in Japan, marking a significant milestone in its development. Ms. Yang concluded: "We are energized by the opportunity to advance SPD8 and potentially add an additional revenue stream to GlycoNex's business, while showcasing our expertise in engineering monoclonal antibodies (mAbs) for a host of disease indications. This includes the continued development of our GNX102 antibody-drug conjugate (ADC) product, which combines a proprietary mAb and cytotoxic drug therapies for the treatment of multiple, high-incidence cancers." About GlycoNex Inc. GlycoNex Inc. is a clinical-stage biotechnology company focused on the development of glycan-directed cancer therapies that effectively inhibit tumor growth while minimizing side effects. GlycoNex possesses a robust pipeline led by GNX102, a humanized monoclonal antibody (mAb) designed to target abnormal sugar molecules in cancer cells. GNX102 has successfully completed Phase 1 clinical trials with data demonstrating excellent safety and promising efficacy. GlycoNex is also advancing a portfolio of antibody-drug conjugates (ADCs) that precisely attack cancer cells while sparing healthy tissue. GlycoNex is headquartered in New Taipei City, Taiwan. For more information, visit . Contact s for GlycoNex, Inc. Investor Relations Tiberend Strategic Advisors, Inc.   David Irish [email protected] Media Relations  Eric Reiss [email protected]   SOURCE GlycoNex, Inc.

临床1期抗体药物偶联物临床结果临床3期

2024-02-04

·药精通Bio

mRNA免疫疗法在恶性脑肿瘤中的应用

IVC2024第四届核酸疫苗与创新型疫苗论坛议程曝光点击图片查看，报名即将截止：（扫码立即领取限量免费参会入场券）免疫疗法改变了癌症治疗的前景，免疫检查点抑制剂 (ICIs)和过继性细胞疗法 (ACT) 在治愈多种癌症类型，特别是在黑色素瘤和血液恶性肿瘤应用方面。然而，实体瘤如胶质母细胞瘤（GBM）和髓母细胞瘤（MB ）疗效仍然有限。当前基于 mRNA 的免疫治疗已成为一个有前途的研究领域，特别是mRNA疫苗与ACT细胞疗法相结合以及负载 mRNA 的树突状细胞 (DC) 疫苗在脑肿瘤临床和临床前研究中的相关性。肿瘤异质性是 GBM 和 MB 的一个标志，导致免疫抵抗和逃逸。在最近的一项研究中显示，经过CAR-T 细胞治疗后 GBM 中肿瘤免疫编辑和抗原靶标丢失。为了克服这些限制，必须研究全谱肿瘤抗原的利用，其中包括新抗原和肿瘤相关抗原（TAA）。疫苗接种策略可以引入多种肿瘤抗原并系统性激活肿瘤抗原特异性T细胞，以增强肿瘤部位的免疫反应。在这方面mRNA疫苗比其他方式表现出独特的优势。首先，mRNA 可以同时递送多种肿瘤抗原，使它们更能抵抗免疫编辑和抗原丢失。其次，mRNA 可以轻松编码全长肿瘤抗原，从而允许APC同时呈递和交叉呈递多个抗原表位。此外，mRNA有潜力诱导更广泛的T细胞反应，而不受HLA类型的限制，多肽的疫苗通常受到这种限制。2024年1月25日，佛罗里达大学研究人员在《Genome Medicine》刊登了一篇“mRNA-based precision targeting of neoantigens and tumor-associated antigens in malignant brain tumors” 研究文章，将预测的肿瘤新抗原与相关抗原的mRNA免疫疗法应用到了脑部肿瘤中，并通过实验验证了该疗法在胶质母细胞瘤（GBM）模型中的效果。抗原预测和肿瘤抗原特异性TOFU mRNA产生a，肿瘤抗原特异性 TOFU mRNA疫苗开发示意图。b，使用开放阅读框架抗原网络( ORAN ) 流程预测KR158-Luc (Kluc)、GL261、NSC 和 Ptch 肿瘤的新抗原或肿瘤相关抗原 (TAAs) 数量。c – f，与Kluc ( c )、GL261 ( d )、NSC ( e ) 和 Ptch ( f ) 肿瘤的总肿瘤 RNA (ttRNA) 相比，TOFU mRNA 文库中肿瘤抗原的富集情况。g – j，在 Kluc ( g )、GL261 ( h )、NSC ( i ) 和 Ptch ( j ) 肿瘤中富集后，TOFU抗原表达的倍数变化增加。K，显示来自人类 GBM 患者肿瘤 RNA 样本的肿瘤特异性和相关基因的富集效率，TOFU 基因表示以蓝色显示，其他基因以橙色显示。首先，研究人员使用了一种名为ORAN（Open Reading Frame Antigen Network）的免疫基因组学流程来预测肿瘤特异性抗原。他们通过对小鼠GBM肿瘤和人类GBM肿瘤的基因表达谱进行分析，预测出了一系列新抗原和肿瘤相关抗原（TAAs），这些抗原可以开发为TOFU mRNA疫苗。TOFU mRNA 疫苗联合 ICI 的治疗效果a ，TOFU 疫苗加抗PD-1 Ab 组合方法的治疗给药时间表；b，在肿瘤植入后第20 天第三次疫苗治疗后 48 小时使用体内发光成像进行 Kluc 肿瘤生长测量。C，用Kluc TOFU mRNA疫苗加抗PD-1 Ab（TOFU DC + PD-1）治疗的 Kluc 荷瘤小鼠和对照治疗（Ctl DCs + IgG、Ctl DCs + PD-1 和 TOFU DCs + IgG）的生存曲线。D，用GL261 TOFU DCs + PD1 组合和对照治疗治疗的 GL261 荷瘤小鼠的生存曲线。接下来，研究人员开发了一种名为TOFU mRNA的免疫疗法。他们使用mRNA技术合成了包含多个肿瘤抗原的mRNA库，并将其注射到小鼠的脾脏细胞中。然后，他们收集这些细胞，通过DCs和PD-1的处理，使其激活T细胞（TOFU-DCs + PD-1）。最后，他们将这些激活的T细胞重新注射到小鼠的体内（TOFU-ACT）。通过实验验证，研究人员发现，接受TOFU-ACT治疗的小鼠在存活率上有显著提高。他们观察到，TOFU-ACT治疗可以增加肿瘤内淋巴细胞的存在，并改变免疫抑制性的肿瘤微环境。此外，TOFU-ACT治疗还可以增加效应/效应记忆CD4 +和CD8 + T细胞的数量，并提高细胞毒性标志物的表达。虽然TOFU-ACT治疗中的T细胞上检测到疲劳标志物的表达，但与未经治疗的小鼠相比，其表达水平较低。TOFU mRNA 疫苗联合 ACT 的治疗效果a，使用 9 Gy TBI 和 HSC 进行宿主预处理后，使用 TOFU 疫苗加 ACT 组合方法进行治疗的时间表。b，在肿瘤植入后第 32 天使用体内发光成像进行 Kluc 肿瘤生长测量。c，用 Kluc TOFU-ACT 治疗和对照治疗的 Kluc 荷瘤小鼠的生存曲线。d，用GL261 TOFU-ACT 和对照治疗治疗的 GL261 荷瘤小鼠的生存曲线。e，来自TOFU-ACT 和对照组的1000 个 CD45 - ve 单细胞的均匀流形近似和投影(UMAP)，这些细胞被分解为8个簇，各个群体中的细胞类型和细胞百分比都有标记。这些结果表明，TOFU mRNA导向的免疫疗法可以增强肿瘤浸润淋巴细胞的数量和功能，从而在治疗小鼠中产生有利的抗肿瘤效果。这种基于mRNA的免疫疗法，通过靶向多个肿瘤抗原来提高GBM患者的生存率。另外，该疗法还可以激活T细胞并改变免疫抑制性的肿瘤微环境。总之，这项研究开发一种针对中枢神经系统恶性肿瘤的mRNA的免疫治疗方法。研究使用了多种模型，包括小鼠胶质母细胞瘤和髓母细胞瘤，通过RNA测序和全外显子测序确定了肿瘤的基因表达谱，并应用了免疫基因组学方法来预测新抗原和肿瘤相关抗原。研究结果显示，通过mRNA疫苗治疗可以提高小鼠的存活率，并且能够激活和增强肿瘤浸润淋巴细胞的效应功能，改变免疫抑制的肿瘤微环境。此外，研究还发现，通过抗PD-1治疗可以减轻T细胞的疲劳状态，进一步提高治疗效果。这些都表明基于mRNA的抗原定向免疫治疗方法可以作为一种个体化和有效的治疗策略，用于治疗侵袭性脑部癌症。原文链接：https://genomemedicine.biomedcentral.com/articles/10.1186/s13073-024-01281-z#Sec31ENDIVC2024第四届核酸疫苗与创新型疫苗论坛招商热线：李欣欣 158 0045 2389戳这里，阅读原文立即领取免费参会入场券

信使RNA免疫疗法疫苗细胞疗法

100 项与 Pro Ab (GlycoNex) 相关的药物交易

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