NGFB

CAMBRIDGE, Mass., July 30, 2024 /PRNewswire/ -- NeuroBo Pharmaceuticals, Inc. (Nasdaq: NRBO), a clinical-stage biotechnology company focused on the transformation of cardiometabolic diseases, today announced the signing of an exclusive license agreement, providing MThera Pharma Co., Ltd. (MTHERA) with the rights to NB-01 for the treatment of painful diabetic neuropathy. The agreement allows MTHERA to conduct research and clinical trials, including, but not limited to, a potential Phase 3 clinical trial in the United States and South Korea, for the future commercialization of NB-01. Financial terms of the agreement were not disclosed. "Finalizing this agreement for NB-01 is an important milestone for NeuroBo, reflecting our stated commitment to out-license our legacy assets in order to focus on the continued clinical development of DA-1726 and DA-1241," stated Hyung Heon Kim, President and Chief Executive Officer of NeuroBo. "MTHERA has extensive experience in manufacturing, quality control and clinical development of natural medicines and botanical drugs, making the company an ideal partner to continue the development of NB-01." "Based on the positive Phase 2 efficacy results for NB-01 for diabetic neuropathy, we intend to develop it as a potential treatment for peripheral diabetes," stated Dr. Mi Won Sohn, Chief Executive Officer of MTHERA. "Utilizing our SyMthomics platform technology, we can clearly identify the mechanism of action and active ingredients of NB-01 to assist in predicting its clinical efficacy as an innovative treatment, with the intent of advancing NB-01 into the next phase of clinical development. We are grateful to NeuroBo for the opportunity to further advance NB-01, and to potentially bring this promising therapy to patients in need." About MThera Pharma MThera Pharma Co., Ltd., headquartered in Seoul, South Korea, is a biopharmaceutical company developing first-in-class botanical drug products using novel, multi-component/multi-target-driven disease-modifying therapies to address diverse etiology and treat chronic incurable diseases such as Parkinson's disease, dementia and inflammatory bowel disease. MThera's platform technology, SyMthomics, consists of its MThera-CODA system (AI based in silico system), a cutting-edge multi-omics integration technology, as well as systems biology and bioinformatics. MThera's standardization technology and advanced CMC technology for raw materials, drug substances and drug products are designed to meet the U.S. Food and Drug Administration requirements for therapeutic consistency. For more information, please visit . About NeuroBo Pharmaceuticals NeuroBo Pharmaceuticals, Inc. is a clinical-stage biotechnology company focused on transforming cardiometabolic diseases. The company is currently developing DA-1726 for the treatment of obesity, and is developing DA-1241 for the treatment of Metabolic Dysfunction-Associated Steatohepatitis (MASH). DA-1726 is a novel oxyntomodulin (OXM) analogue that functions as a glucagon-like peptide-1 receptor (GLP1R) and glucagon receptor (GCGR) dual agonist. OXM is a naturally-occurring gut hormone that activates GLP1R and GCGR, thereby decreasing food intake while increasing energy expenditure, thus potentially resulting in superior body weight loss compared to selective GLP1R agonists. DA-1241 is a novel G-protein-coupled receptor 119 (GPR119) agonist that promotes the release of key gut peptides GLP-1, GIP, and PYY. In pre-clinical studies, DA-1241 demonstrated a positive effect on liver inflammation, lipid metabolism, weight loss, and glucose metabolism, reducing hepatic steatosis, hepatic inflammation, and liver fibrosis, while also improving glucose control. For more information, please visit . Forward Looking Statements Certain statements in this press release may be considered forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995. Words such as "believes", "expects", "anticipates", "may", "will", "should", "seeks", "approximately", "intends", "projects", "plans", "estimates" or the negative of these words or other comparable terminology (as well as other words or expressions referencing future events, conditions or circumstances) are intended to identify forward-looking statements. Forward-looking statements are predictions, projections and other statements about future events that are based on current expectations and assumptions and, as a result, are subject to risks and uncertainties. Many factors could cause actual future events to differ materially from the forward-looking statements in this press release, including, without limitation, those risks associated with NeuroBo's ability to execute on its commercial strategy; the timeline for regulatory submissions; the ability to obtain regulatory approval through the development steps of NeuroBo's current and future product candidates, the ability to realize the benefits of the license agreement with Dong-A ST Co. Ltd., including the impact on future financial and operating results of NeuroBo; the cooperation of NeuroBo's contract manufacturers, clinical study partners and others involved in the development of NeuroBo's current and future product candidates; potential negative interactions between NeuroBo's product candidates and any other products with which they are combined for treatment; NeuroBo's ability to initiate and complete clinical trials on a timely basis; NeuroBo's ability to recruit subjects for its clinical trials; whether NeuroBo receives results from NeuroBo's clinical trials that are consistent with the results of pre-clinical and previous clinical trials; impact of costs related to the license agreement, known and unknown, including costs of any litigation or regulatory actions relating to the license agreement; the effects of changes in applicable laws or regulations; the effects of changes to NeuroBo's stock price on the terms of the license agreement and any future fundraising; and other risks and uncertainties described in NeuroBo's filings with the Securities and Exchange Commission, including NeuroBo's most recent Annual Report on Form 10-K. Forward-looking statements speak only as of the date when made. NeuroBo does not assume any obligation to publicly update or revise any forward-looking statements, whether as a result of new information, future events or otherwise, except as required by law. Contacts: NeuroBo Pharmaceuticals Marshall H. Woodworth Chief Financial Officer +1-857-299-1033 [email protected] Rx Communications Group Michael Miller +1 917-633-6086 [email protected] SOURCE NeuroBo Pharmaceuticals, Inc.

重组人源神经生长因子介绍神经生长因子（Nerve Growth Factor, NGF）是最早发现和最典型的神经营养因子，同时也是影响神经系统最重要的生物活性分子之一。人神经生长因子（Human Nerve Growth Factor, hNGF）是在人体中发现的一种对正常神经细胞具有营养作用，同时对损伤神经修复机能起调节作用的生物活性因子。NGF不仅可以维持交感神经和感觉神经的生存和功能，促进神经细胞的分化，决定轴突的伸展方向，还能对促进大脑的发育、神经系统的生长，以及损伤神经的再生和功能恢复具有决定性作用（图1）。图1 生长因子作用机制图示完整的hNGF分子由α、β、γ三个亚基组成，其中β亚基（β-NGF）是唯一具有生物学活性的亚基。β亚单位通过非共价链结合而构成活性二聚体，其中，6个半胱氨酸残基之间由二硫键相连，若二硫键结构不正确，NGF将丧失其生物活性。β-NGF对中枢及周围神经元的发育、分化、生长、再生和功能特性的表达，均具有重要的调控作用。目前，NGF 及其受体已成为临床镇痛药物开发的重要靶点，β-NGF已被开发成治疗外周神经损伤的药物，用于治疗神经中毒、周围神经损伤、糖尿病性周围神经病变、老年痴呆病、帕金森氏病、面神经炎，以及神经损伤（包括脊髓损伤、高位截瘫、断指再植、脑损伤等中枢及外围神经系统的损伤）等神经系统疾病。β-NGF也是目前唯一可应用于临床的神经系统蛋白质因子（图2）。图2 神经生长因子（NGF）在调节中枢神经系统疼痛信号中的作用[1]目前商品化的NGF为鼠源性神经生长因子（mNGF），且mNGF与hNGF基因序列相比高度同源（>90％），从下颌下腺分离的mNGF和从胎盘分离的hNGF已成功用于临床，主要用于治疗视神经损伤。除此之外，NGF也对毒性神经病变、外周神经病变和面部神经病变等疾病有疗效。然而，这类动物源性蛋白制品常常具有较高的免疫原性，容易引起人体内的抗原反应，甚至会危及生命。而天然hNGF分布于人体的脑、神经节、虹膜、心脏、脾和胎盘等组织，数量有限，且其中hNGF含量低，分离纯化极其困难，通常无法直接提取获得。因此，利用基因工程技术制备重组人神经生长因子，是解决上述问题的一个主要途径。重组人源神经生长因子的表达利用基因工程技术，以微生物为载体，大批量生产外源蛋白，具有广阔的应用前景。到目前为止，已有多项研究利用哺乳动物细胞、昆虫细胞和大肠杆菌等系统，成功表达出人β-NGF。在上述表达系统中，人们对大肠杆菌的遗传背景和生化特性了解得相对透彻，大肠杆菌本身具备容易操作、生长迅速、营养需求简单等优点。同时，大肠杆菌表达系统具有表达水平高、生产成本低和便于工业化生产等优势，成为制备重组人神经生长因子的首选。但是这一系统本身也存在着若干缺陷：①缺少真核生物的蛋白翻译后修饰和加工过程，如剪切、糖基化和二硫键形成等；②表达的蛋白多以包涵体形式存在，需要经过复杂的复性步骤才能恢复蛋白构象和活性；③较多的背景杂蛋白，纯化工艺复杂；④真核生物和原核生物所偏好的密码子存在差异，部分真核基因在大肠杆菌表达系统中表达量极低，甚至不表达。有研究发现，人β-NGF在大肠杆菌中的表达量不高，而且生物活性极低，主要是由于β-NGF基因及其蛋白的结构特殊，导致其难以在大肠杆菌系统中有效表达。一方面，β-NGF基因中使用了较多的大肠杆菌稀有密码子，导致β-NGF基因在大肠杆菌中表达量不高。另一方面，β-NGF蛋白难以在缺乏翻译后加工修饰系统的大肠杆菌中形成正确的结构，最终导致基于大肠杆菌表达系统的β-NGF蛋白生物活性极低，甚至没有活性。基于上述问题，有研究报道了一种重组人神经生长因子β亚基基因，并提供了利用大肠杆菌表达系统制备重组人神经生长因子的方法。该序列位于人神经生长因子β亚基成熟肽基因序列上游，两序列由肠激酶切割位点连接。分子伴侣可帮助人神经生长因子β亚基（以包涵体形式表达）在体外形成正确的构象，并且能在复性后通过肠激酶的切割作用被去除，从而释放出与天然β-NGF氨基酸序列完全一致，且具有较高生物学活性的成熟肽。中国仓鼠卵巢（CHO）细胞作为使用最为广泛的哺乳动物细胞表达系统之一，常用于重组NGF的表达研究，重组NGF被表达并分泌到细胞外，以游离形式（与天然形态类似）存在于上清液中。相较于大肠杆菌系统，CHO系统不仅具有良好的翻译后修饰能力，还支持大规模生产NGF。在真核细胞中表达，更有利于得到活性较高和更接近自然状态的β-NGF蛋白，具体优点如下：①能够加工表达的蛋白质，如空间构型的折叠、糖基化和磷酸化等；②可识别和除去外源基因中的内含子，进一步剪切加工为成熟的mRNA；③转染目的基因的真核细胞可冻存，复苏后能稳定表达活性目的蛋白。目前，使用真核系统大规模表达、纯化NGF已有成熟工艺，而用原核系统表达的重组人源人神经生长因子（rhNGF）多因不能正确折叠而使生物学活性只有mNGF五十分之一，通常首选真核表达系统来克服蛋白错误折叠等问题。近年来，研究人员通过构建具有前导序列的NGF来帮助其体外复性，虽然复性率有所提高，但产物需经限制酶切后才能获得成熟NGF，后续处理难度相对增加。综合两种表达体系可知，哺乳动物表达系统具有分泌正确装配、折叠与糖基化等特点，更适合表达NGF。韩晓枫团队在COS-7细胞中对NGF进行了瞬时表达，结果表明表达产物具有生物活性，但是无法实现NGF的稳定、长期表达，建议只作为瞬时表达宿主细胞。而CHO细胞具有利于二硫键形成的细胞环境。姜静团队对比了来源于E .coli和CHO表达系统的rhβ‑NGF，发现采用原核系统表达的NGF多以不溶性包含体形式存在（可能是游离蛋白而非跨膜蛋白），这为NGF能否在细胞表面表达存在很大的不确定性本是否。相比之下，通过将NGF的核酸序列与适当的铰链区和跨膜区的核酸序列串联后导入CHO细胞，就可以在重组CHO细胞表面上稳定表达NGF，使NGF存在于细胞表面而非分泌到培养液中，同时确保NGF经过恰当的翻译后加工（包括正确折叠、糖基化等），从而保持天然构象和活性。重组人源神经生长因子的纯化来源于CHO系统的rhNGF中往往含有变异体，如细胞内分泌过程中翻译后修饰、分泌后氨基酸残基侧链发生化学反应，或肽链降解而形成的一系列蛋白。rhNGF在体内以前体形式合成，由于弗林蛋白酶或激素酶原转化酶加工不完全，会产生完全或部分前体。除前体外，由于真核细胞特性，还会产生诸如N端截断（由于翻译后加工导致N端部分氨基酸缺失的序列分子）、氧化、脱酰胺、异构、C末端截断和异常（由于翻译后加工导致结构异常、二硫键异常、异常氧化等变体的统称）等变异体。常规的层析方法能够除去许多工艺相关杂质（如宿主细胞蛋白及核酸等），但是很难除作为产品相关杂质的rhNGF变异体。这些变异体通常与成熟rhNGF一同产生，两者理化性质相近，为大规模纯化增加了难度。目前纯化rhNGF的报道主要包括：1. 史权威团队采用阳离子交换及分子筛两步方法制备了纯度大于98%的rhNGF，但该阳离子交换步骤仅用于清除宿主细胞蛋白等过程杂质；2. 专利CN106478801A采用阳离子交换及疏水层析制备了纯度大于99%的rhNGF，该阳离子交换主要用于捕获产品，目的是清除宿主细胞蛋白等工艺相关杂质；3. 专利CN1268639C利用高效阳离子交换，以线性洗脱方式分离rhNGF氧化、异构、脱酰胺等变体；4. 还有研究通过分析rhNGF及其变异体的理化性质，发现在弱阳离子交换色谱条件下，N端截断及异常变体具有更低的等电点，可采用阶段升高电导率的方法清除变体。想要从包括发酵液、裂解的细菌或哺乳动物细胞中大规模纯化NGF，制备性分离获得具有药物纯度和高产量（基本上不含变体）的rhNGF满足临床需求难度较高，需要一种有效的方法来选择性地从其变体和其它分子、以及pI高的其它多肽中分离出NGF。有研究报道了一种使用疏水作用层析（HIC）来纯化NGF的方法，可以将成分不同或错折叠形成的分子与目标蛋白分开，除去在疏水性上不符合要求的变体，包括经部分加工的前体序列、含有糖基化成熟或前体的形式以及错误折叠和部分折叠的变体。除此以外，另一种制备性阳离子交换层析纯化NGF的方法也被广泛应用，该方法可将电荷变体（如氧化、异天冬氨酸和脱酰胺形式）从成熟的NGF蛋白中分离出来，采用HIC和阳离子交换层析制备的NGF产物基本不含加工和电荷变体。小编总结综上所述，未来有关NGF的研究及应用发展可能主要在如下几个方向：首先，随着基因工程制备rhNGF技术工艺的逐步成熟，最终可实现规模化生产水平。rhNGF因其特性和较高的分子同源性，生产规模不受原料限制，且无动物来源的致病病毒污染，具有更大的发展潜力。我们将重点研究如何优化发酵和纯化条件，获得稳定高效的表达途径，最终高效实现规模化生产。其次，近期文献资料显示，国外相关研究的热点逐渐转向NGF的二肽模拟物，部分学者认为，NGF的二肽模拟物可作为治疗神经损伤的候选药物，具有更加广阔的发展和应用前景。合作意向y目前，甲贝医药针对NGF项目已建立完整的构建、表达、纯化及制剂开发流程，欢迎感兴趣的合作伙伴前来沟通，共同开发。图3 甲贝医药的一站式表达、纯化及制剂开发流程对接人：徐先生，18616852990 微信号：参考来源：[1] https://mosmedpreparaty.ru/news/14000本文不具有任何投资建议和医学意见点击“阅读原文”，填写问卷，前10名有奖~