Abatacept

蛋白质的糖基化是指寡糖链通过共价键与肽链结合，是蛋白质重要的翻译后加工修饰之一。对蛋白质的结构稳定性、血清半衰期、生物活性及溶解度等关键属性都可能具有重要影响，通常是重组蛋白/抗体类生物制品的关键质量属性（CQA）。 酸性单糖唾液酸通常位于修饰蛋白的寡糖链末端，唾液酸化对重组蛋白类药物的药代动力学具有重要影响，可降低蛋白质在体内被清除的速率，还参与细胞识别、信号传导等生物过程。 今天为大家介绍一种增强CHO细胞培养物中Fc融合蛋白的唾液酸化的有效方法。 人类蛋白质糖链的唾液酸通常为α2,6-唾液酸和α2,3-唾液酸。由于编码α2,6- ST的st6gal1基因启动子被甲基化失活，做为重组蛋白类药物生产重要宿主的CHO细胞无法产生α2,6-唾液酸，因此仅能对其表达的蛋白质进行α2,3-唾液酸化修饰。 对CHO细胞来源的重组蛋白唾液酸连接进行双重处理可使其更接近人类的蛋白质，从而减少或避免治疗性蛋白引发的机体的免疫反应。 目前许多研究聚焦于通过上调唾液酸转移酶活性和下调唾液酸酶活性来最大化唾液酸含量。其中最直接的方法是通过调整培养基组分来实现唾液酸水平的提升。 糖皮质激素（GCs）一类类固醇激素。因与糖皮质激素受体（GRs）可特异性结合并激活其功能，常被用来调控细胞代谢、增殖、分泌以及糖蛋白糖基化等生理过程。 今天挑选常见的六种GCs（图1），分别是氢化可的松（Hyd）、皮质酮（Ccs）、地塞米松（Dex）、 倍他米松（Bet）、 泼尼松龙（Prl） 和 泼尼松（Prd），评估GCs作为培养基添加剂在提升CHO细胞培养中重组蛋白质量方面的应用价值。 图1：用作CHO细胞培养物添加剂的GCs的化学结构 1 材料与方法 细胞：CHO DUKX-B11，在ST6Gal1中过表达并表达α2,6-ST的CTLA4-Ig 测试添加剂：Hyd、Ccs、Dex、Bet、Prl和Prd 浓度范围：0、0.5、1、10 和 20 mg/L 培养模式：CHO细胞以3×106个细胞/mL的密度接种至工作体积为20 mL的125 mL锥形瓶中，置于在 37 ℃、100 rpm 、 5% CO2 摇床中培养。从第1天到第8天，每24小时取样一次。监测活细胞密度（VCD）和活率。 2 结果 GCs对CHO细胞培养性能的影响 图2：补充GC培养物的活细胞密度（VCD）分布曲线 除20 mg/L浓度外，添加GC对VCD的影响与对照组相比无显著差异。 图3：补充GC培养物的细胞活力曲线 在所有测试的GC浓度下，CHO细胞培养物的活率均未受影响。 图4：GC对CHO细胞培养中（A） IVCD和（B）滴度的影响 通过ELISA定量检测CTLA4-Ig水平发现，GCs对CTLA4-Ig的产生具有浓度相关性的负面影响（图4B）。20 mg/L Ccs下CTLA4-Ig的产量降低了47%，但10 mg/L Ccs培养的CTLA4-Ig滴度则与对照相当。 这些数据表明，糖皮质激素的使用会导致CTLA4-Ig的生成减少，但在CHO细胞培养体系中，糖皮质激素的使用浓度可达到10 mg/L。 结合此前研究报道指出，糖皮质激素会抑制哺乳动物细胞的葡萄糖转运和谷氨酰胺摄取功能。CHO细胞培养物中添加GC后，细胞生长和重组蛋白产量降低与葡萄糖和谷氨酰胺摄取受抑制有关。 盐酸水解α2,3-和α2,6-唾液酸，而唾液酸酶S仅能水解α2,3-唾液酸。如图5所示，添加10 mg/L浓度的Ccs可显著提升总唾液酸含量。与对照组相比，含量变化幅度为1.16倍。实验数据表明，在CHO细胞培养体系中添加Ccs等某些糖基化辅酶， 能有效提升唾液酸化水平。 CTLA4-Ig中唾液酸的相对丰度   图5：α2,3-唾液酸含量（A）与总唾液酸含量（B）的分析结果 细胞内唾液酸转移酶在将CMP-唾液酸递送至糖链残基的过程中起着关键作用。因此，提高该酶的活性可有效调控重组蛋白的唾液酸化水平。如图6所示，与对照组相比，添加10 mg/L Ccs后，细胞内唾液酸转移酶活性显著增加。 细胞内唾液酸转移酶活性 图6：第6天的相对细胞内唾液酸转移酶活性 CHO 细胞的内源性唾液酸酶，可在细胞培养过程中在培养上清液中积累，导致分泌至培养上清中糖蛋白糖链末端唾液酸被移除。因此，唾液酸酶的下调可提高糖蛋白的唾液酸化程度。如图7所示，在培养第6天，补充有10 mg/L Ccs 的细胞外唾液酸酶活性比对照组降低了25%。 细胞外唾液酸酶活性 图7：第6天细胞外唾液酸酶活性分析 3 结论 本文描述了在表达α2,3和α2,6-ST的CHO细胞培养液中，添加不同种类的GCs对细胞培养性能和Fc-融合蛋白唾液酸化的影响。尽管额外添加GCs可对生产效率产生负面影响，但可提升重组蛋白的唾液酸总量。其中，Ccs对提升Fc融合蛋白总唾液酸化水平最为显著，包括具有类人唾液酸化特征的α2,6-唾液酸。 唾液酸含量的增加主要归因于两个因素：细胞内唾液酸转移酶活性上调与细胞外唾液酸酶活性下调。唾液酸转移酶活性增强后，能更有效地将唾液酸转移到末端半乳糖残基上。此外，在Ccs处理条件下，由于唾液酸酶活性降低，Fc融合蛋白上的唾液酸裂解过程受到抑制。 本文介绍了一种提高CHO细胞培养物中重组蛋白质量的有效策略，通过将Ccs作为细胞培养添加剂，可有效增强细胞内唾液酸转移酶活性，并抑制细胞外唾液酸酶活性，从而提高唾液酸化水平，最终实现提升CHO细胞重组蛋白质量的目标。 参考文献： Utilization of Glucocorticoids as Additives for Enhanced Sialylation of Fc-fusion Protein in CHO Cell Cultures[J]. Biotechnology and Bioprocess Engineering 26: 286-293

HOUSTON, Aug. 12, 2025 /PRNewswire/ -- Coya Therapeutics, Inc. (NASDAQ: COYA) ("Coya" or the "Company"), a clinical-stage biotechnology company focused on developing biologics that enhance regulatory T cell (Treg) function in patients with neurodegenerative disorders, provides a corporate update and announces its financial results for the quarter ended June 30, 2025. Recent Corporate Highlights Submitted to FDA on June 30, 2025, additional nonclinical data to support the start of the COYA 302 Phase 2 trial in patients with ALS Published results of scientific research linking inflammation and oxidative stress to the progression of Parkinson's Disease (PD). Study demonstrated correlation between peripheral pro-inflammatory mechanisms, particularly monocytes and oxidative stress, in the progression and severity PD. It was published in the scientific journal Frontiers of Immunology Announced U.S. patent (US 12,312,389 B2) for a stable ready-to-use liquid IL-2 formulation; Coya holds exclusive in-vivo rights across multiple indications Upcoming Expected Catalysts for 2025 Decision from the US FDA on the IND for COYA 302 in ALS, expected by or before August 29, 2025 Upon IND acceptance and first patient dosing of COYA 302 in ALS, Coya will receive milestone payments of $8.4 million from strategic partner, Dr. Reddy's Laboratories (DRL) ALS Biomarker data. Publication of longitudinal data on Neurofilament Light Chain (NfL) and oxidative stress markers in tracking progression and survival in patients with ALS Report new proteomics data from the completed investigator-initiated, 21-week, double-blind, placebo-controlled, exploratory Phase 2 study of low-dose interleukin-2 (LD IL-2) in patients with Alzheimer's disease (AD) Top-line clinical data release for an investigator-initiated trial combining LD IL-2 + CTLA4-Ig in patients with Frontotemporal Dementia (FTD) Top-line data for COYA 303 in an in-vivo inflammatory animal model Filing of IND for the COYA-302 Phase 2 trial in patients with FTD* (*Clinical trial will be initiated after FDA IND approval for FTD) Coya's Chief Executive Officer Arun Swaminathan, Ph.D. commented, "As we look ahead, we are focused on several key catalysts, including a decision from the FDA on our IND application for COYA 302 in ALS, anticipated by the end of August. We are encouraged by the growing body of data supporting our regulatory T cell enhancement approach, including the recent findings from the investigator-initiated trial in FTD and promising pre-clinical results with COYA 303. These advances strengthen our confidence in the potential of COYA 302 in neurodegenerative diseases like ALS and FTD." Coya's Chief Medical Officer Dr. Fred Grossman commented, "We look forward to the FDA's decision on our IND submission for COYA 302 in ALS, which represents a significant milestone for our lead program. Pending clearance, we are prepared to initiate our controlled Phase 2 clinical trial in ALS. We also plan to submit an IND for FTD by the end of the year." Financial Results As of June 30, 2025, Coya had cash and cash equivalents of $29.8 million. Collaboration revenues were $0.2 million for the three months ended June 30, 2025, compared to $3.4 million for the three months ended June 30, 2024, a change of approximately $3.3 million. The decrease was primarily due to the immediate recognition of license revenue upon executing the First Amendment to the DRL Development Agreement during the three months ended June 30, 2024. Research and development expenses were $3.7 million for the three months ended June 30, 2025, compared to $4.6 million for the three months ended June 30, 2024, a decrease of approximately $0.9 million. The decrease was primarily due to a $1.2 million decrease in our preclinical expenses primarily due to our preclinical work on COYA 302 in ALS, partially offset by a $0.2 million increase in internal research and development expenses, and a $0.1 million increase in sponsored research. General and administrative expenses were $2.9 million for the three months ended June 30, 2025, and $2.1 million for the three months ended June 30, 2024, an increase e of approximately $0.8 million. The increase was primarily due to a $0.3 million increase in stock-based compensation, a $0.4 million increase in professional services, and a $0.1 million increase in investor relation related expenses. Net loss was $6.1 million for the three months ended June 30, 2025, compared to net loss of $2.9 million for the three months ended June 30, 2024. About COYA 302 COYA 302 is an investigational and proprietary biologic combination therapy with a dual immunomodulatory mechanism of action intended to enhance the anti-inflammatory function of regulatory T cells (Tregs) and suppress the inflammation produced by activated monocytes and macrophages. COYA 302 comprises proprietary low dose interleukin-2 (LD IL-2) and CTLA-4 Ig and is being developed for subcutaneous administration for the treatment of patients with ALS. These mechanisms may have additive or synergistic effects. COYA 302 is an investigational product not yet approved by the FDA or any other regulatory agency. About COYA 303 COYA 303 is an investigational biologic combination of COYA 301 and a glucagon-like-peptide-1 receptor agonist (GLP-1 RA) designed for subcutaneous administration. In a preclinical study, COYA 303 exhibited a dual immunomodulatory mechanism of action resulting in an additive/synergistic anti-inflammatory effect, which the Company believes was due to increased Treg function and suppressed pro-inflammatory myeloid cells and responder T cells. About Coya Therapeutics, Inc. Headquartered in Houston, TX, Coya Therapeutics, Inc. (Nasdaq: COYA) is a clinical-stage biotechnology company developing proprietary treatments focused on the biology and potential therapeutic advantages of regulatory T cells ("Tregs") to target systemic inflammation and neuroinflammation. Dysfunctional Tregs underlie numerous conditions, including neurodegenerative, metabolic, and autoimmune diseases. This cellular dysfunction may lead to sustained inflammation and oxidative stress resulting in lack of homeostasis of the immune system. Coya's investigational product candidate pipeline leverages multiple therapeutic modalities aimed at restoring the anti-inflammatory and immunomodulatory functions of Tregs. Coya's therapeutic platforms include Treg-enhancing biologics, Treg-derived exosomes, and autologous Treg cell therapy. For more information about Coya, please visit Forward-Looking Statements This press release contains "forward-looking" statements that are based on our management's beliefs and assumptions and on information currently available to management. Forward-looking statements include all statements other than statements of historical fact contained in this presentation, including information concerning our current and future financial performance, business plans and objectives, current and future clinical and preclinical development activities, timing and success of our ongoing and planned clinical trials and related data, the timing of announcements, updates and results of our clinical trials and related data, our ability to obtain and maintain regulatory approval, the potential therapeutic benefits and economic value of our product candidates, competitive position, industry environment and potential market opportunities. The words "believe," "may," "will," "estimate," "continue," "anticipate," "intend," "expect," and similar expressions are intended to identify forward-looking statements. Forward-looking statements are subject to known and unknown risks, uncertainties, assumptions and other factors including, but not limited to, those related to risks associated with the impact of COVID-19; the success, cost and timing of our product candidate development activities and ongoing and planned clinical trials; our plans to develop and commercialize targeted therapeutics; the progress of patient enrollment and dosing in our preclinical or clinical trials; the ability of our product candidates to achieve applicable endpoints in the clinical trials; the safety profile of our product candidates; the potential for data from our clinical trials to support a marketing application, as well as the timing of these events; our ability to obtain funding for our operations; development and commercialization of our product candidates; the timing of and our ability to obtain and maintain regulatory approvals; the rate and degree of market acceptance and clinical utility of our product candidates; the size and growth potential of the markets for our product candidates, and our ability to serve those markets; our commercialization, marketing and manufacturing capabilities and strategy; future agreements with third parties in connection with the commercialization of our product candidates; our expectations regarding our ability to obtain and maintain intellectual property protection; our dependence on third party manufacturers; the success of competing therapies or products that are or may become available; our ability to attract and retain key scientific or management personnel; our ability to identify additional product candidates with significant commercial potential consistent with our commercial objectives; and our estimates regarding expenses, future revenue, capital requirements and needs for additional financing. We have based these forward-looking statements largely on our current expectations and projections about future events and trends that we believe may affect our financial condition, results of operations, business strategy, short-term and long-term business operations and objectives, and financial needs. Moreover, we operate in a very competitive and rapidly changing environment, and new risks may emerge from time to time. It is not possible for our management to predict all risks, nor can we assess the impact of all factors on our business or the extent to which any factor, or combination of factors, may cause actual results to differ materially from those contained in any forward-looking statements we may make. In light of these risks, uncertainties and assumptions, the forward-looking events and circumstances discussed herein may not occur and actual results could differ materially and adversely from those anticipated or implied in the forward-looking statements. Although our management believes that the expectations reflected in our forward-looking statements are reasonable, we cannot guarantee that the future results, levels of activity, performance or events and circumstances described in the forward-looking statements will be achieved or occur. We undertake no obligation to publicly update any forward-looking statements, whether written or oral, that may be made from time to time, whether as a result of new information, future developments or otherwise. Investor Contact David Snyder, CFO [email protected] astr partners Matthew Beck [email protected] 917-415-1750 Media Contacts Russo Partners Olipriya Das [email protected] 646-942-5588 Matthew Purcell [email protected] 646-942-5595 SOURCE Coya Therapeutics, Inc. WANT YOUR COMPANY'S NEWS FEATURED ON PRNEWSWIRE.COM? 440k+ Newsrooms & Influencers 9k+ Digital Media Outlets 270k+ Journalists Opted In GET STARTED