CN106957359A - Fgf1突变体及其医药应用 - Google Patents
Fgf1突变体及其医药应用 Download PDFInfo
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Abstract
本发明提供了FGF1突变体，其氨基酸序列在SEQ ID NO：1的基础上具有Lys127、Lys128和/或Lys133突变，其减少FGF1促细胞分裂和增殖副作用，但是具有降糖等活性。
Description
技术领域
本发明属于蛋白质技术领域，具体而言，本发明涉及FGF1突变体，其减少FGF1促细胞分裂和增殖副作用，但是具有降糖等活性。
背景技术
成纤维细胞生长因子（Fibroblast Growth Factor，FGF）中共有18个家族成员，被分为5个旁分泌和1个内分泌亚家族。FGF作用的发挥则是通过与细胞表面具有酪氨酸激酶活性的FGF受体（FGF Receptor， FGFR）相互作用进而引起受体的二聚化并进一步激活下游级联信号来实现的。
其中，FGF1可以对db/db, ob/ob等2型糖尿病鼠模型产生重要的降糖和提高胰岛素敏感的功能，而且即使大剂量也不会造成低血糖的产生（可参见Nature Review DrugDiscovery, 2016; Nature Medicine, 2016; Proc Natl Acad Sci U S A, 2016;Diabetes, 2016; Nature, 2015，等）。
然而，FGF1作为一个旁分泌蛋白，它通过和肝素以及FGFR相互作用形成紧密的三元复合物进而启动一系列下游复杂的信号通路，其中就包括促细胞分裂和增殖的效应，因此，频繁的注射FGF1无疑会在体内存在诱导增生甚至肿瘤产生的隐患。
本发明人为此经过长期研究，令人意外地发现了一种FGF1肝素结合位点突变体（简称为FGF1△HS），其无需敲除整个肝素结合位点，而仅仅需要引入有限的突变，即可使得FGF1△HS仅能与FGFR形成较弱的二元复合物即可启动代谢调控的信号通路，而基本丧失了促细胞分裂和增殖的功能，同时，FGF1△HS又保持了降糖等药理作用。
发明内容
本发明要解决的技术问题在于提供新的FGF1突变体，其基本消除了野生型FGF1的促细胞分裂和增殖的能力，但是仍旧保留了FGF1的降血糖等功能，从而可以长期安全地给药。另外，本发明还提供了该突变体的编码基因、表达载体和宿主细胞以及医药用途等。
具体而言，在第一方面，本发明提供了FGF1突变体，其氨基酸序列在SEQ ID NO：1的基础上（仅）具有Lys127、Lys128和/或Lys133突变。本发明第一方面的突变体减少（优选消除）野生型FGF1（其氨基酸序列如SEQ ID NO：1所示）的促细胞分裂和增殖的能力，但是仍旧保留了降血糖等功能。
在本文中，突变的表示采用本领域技术人员所熟知的方式。例如，Lys127表示对第127位上的Lys进行突变；而Lys128Gln表示对第128位上的Lys进行突变，突变为Gln。突变可以是添加、缺失和/或取代。本领域技术人员知晓，通过改变已知多肽的编码基因序列并将其导入表达载体，可以制备出取代、添加和/或缺失了氨基酸残基的多肽或蛋白质，这些方法广泛记载于《分子克隆实验指南》等本领域公知的文献中。在本发明的具体实施方式中，突变是取代。
优选本发明第一方面的突变体（仅）具有Lys127Asp、Lys128Gln和/或Lys133Val突变。在本发明的具体实施方式中，该突变体（仅）具有Lys127Asp、Lys128Gln和Lys133Val突变。
在第二方面，本发明提供了编码本发明第一方面的突变体的多核苷酸。在本文中，多核苷酸可以是DNA形式，也可以是RNA形式，优选DNA形式。DNA形式包括天然cDNA和人工合成的cDNA，DNA可以是编码链或模板链。通过常规技术，如PCR方法、重组法或人工合成的方法，本领域技术人员可以很容易获得编码本发明的突变体的核酸分子多核苷酸或其片段。这些序列一旦获得，就可以将其克隆入载体，再转化或转染入相应的细胞，然后通过常规的宿主细胞进行增殖，从中分离得到大量的核酸分子。
在第三方面，本发明提供了一种载体，其含有本发明第二个方面所述的多核苷酸。在本文中，载体包括表达载体和克隆载体，是指本领域中常用的细菌质粒、粘粒、噬菌粒、酵母质粒、植物细胞病毒、动物病毒及其它各种病毒载体。本发明中适用的载体包括但不限于：在细菌中表达用的载体（原核表达载体）、在酵母中表达用的载体（如毕赤酵母载体、汉逊酵母载体等）、在昆虫细胞中表达的杆状病毒载体、在哺乳动物细胞中表达用的载体（痘苗病毒载体、逆转录病毒载体、腺病毒载体、腺伴病毒载体等）、在植物中表达用的植物病毒载体以及在哺乳动物乳腺中表达用的各种载体。总之，只要能在宿主细胞中稳定复制，任何质粒和载体都可使用。优选表达载体包含选择标记基因，如细菌的氨苄青霉素抗性基因、四环素抗性基因、卡那霉素抗性基因、链霉素抗性基因、氯霉素抗性基因；酵母菌的新霉素抗性基因、Zeocin抗性基因，酵母菌的缺陷选择标志，如His, Leu, Trp等；真核细胞的新霉素抗性基因、Zeocin抗性基因、二氢叶酸还原酶基因及荧光蛋白标记基因等。在本发明的具体实施方式中，使用的是已经商品化的pET质粒。
在第四方面，本发明提供了一种细胞，如宿主细胞，其含有本发明第二个方面所述的多核苷酸。本发明的细胞可以含有本发明第三方面的载体，也可以用本发明第二方面的多核苷酸转化或转染。宿主细胞可以是原核细胞，也可以是真核细胞，如，细菌细胞、酵母细胞、植物细胞、昆虫细胞、哺乳动物细胞等。宿主细胞在转化或转染含本发明所述编码融合蛋白的基因序列后，即构成工程化细胞或细胞株，可用于生产所需融合蛋白。本领域技术人员能够恰当地选择适当的载体、宿主细胞，并熟知如何将载体高效地转化或转染入宿主细胞中，所用方法包括但不限于：氯化钙法、电穿孔法用于细菌细胞，电穿孔法和原生质体融合法用于酵母细胞，脂质体包裹、磷酸钙共沉淀、电融合法以及显微注射法用于哺乳动物细胞等真核细胞。优选本发明的宿主细胞是大肠杆菌BL21（DE3）。
在第五方面，本发明提供了药物组合物，其包含本发明第一方面的突变体以及药学上可接受的载体。本发明的药物组合物能治疗糖尿病、降血糖、治疗或预防肥胖症、抗炎或者用于降低炎性细胞因子水平，并且其减少（甚至消除）FGF1的促细胞增殖的副作用。因而，本发明的药物组合物能够长期安全地给药。在本文中，药学上可接受的载体指无毒的填充剂、稳定剂、稀释剂、佐剂或其他制剂辅料。根据本领域的公知技术，可以根据治疗目的、给药途径的需要将药物组合物制成各种剂型，优选该组合物为单位剂量形式，如冻干剂、片剂、胶囊、粉剂、乳液剂、注射剂或喷雾剂，更优选该药物组合物为注射剂型，如冻干粉针剂。也优选该药物组合物是液体制剂，如可以包含缓冲液（如磷酸盐缓冲液、Tris-HCl缓冲液）。
在第六方面，本发明提供了本发明第一方面的突变体在制备减少FGF1促细胞分裂和增殖副作用的药物中的应用。野生型FGF1具有促细胞分裂和增殖的作用，这容易导致增生，乃至癌变。而本发明的突变体及其药物能够显著降低（甚至基本消除）这样的副作用，从而能够长期安全地给药。
本发明的突变体及其药物基本保留了FGF1的降糖的功能。因此，优选在本发明第六方面的应用中，药物用于降血糖。也优选在本发明第六方面的应用中，药物用于治疗糖尿病，尤其是2型糖尿病。
本发明的突变体及其药物基本保留了FGF1的降脂的功能。因此，优选在本发明第六方面的应用中，药物用于降血脂（如，甘油三酯）。也优选在本发明第六方面的应用中，药物用于治疗或预防肥胖症。
本发明的突变体及其药物还具有抗炎的作用，能够用于降低一些炎性细胞因子的水平。因此，优选在本发明第六方面的应用中，药物用于抗炎。也优选在本发明第六方面的应用中，药物用于降低炎性细胞因子（如，IL-6和/或TNF-α）水平。
本发明的有益效果在于：本发明的突变体基本丧失了促细胞分裂和增殖的功能，同时又保持了降糖等药理作用，所以比已经成药的FGF1更具安全用药的前景，基本保留FGF1的整体结构（并没有缺失整个HS结合区），所以可以尽可能地利用现有FGF1的基因工程生产体系（包括载体、细胞和表达条件），方便产业化。
本发明引用了公开文献，这些文献是为了更清楚地描述本发明，它们的全文内容均纳入本文进行参考，就好像它们的全文已经在本文中重复叙述过一样。
为了便于理解，以下将通过具体的实施例对本发明进行详细地描述。需要特别指出的是，这些描述仅仅是示例性的描述，并不构成对本发明范围的限制。依据本说明书的论述，本发明的许多变化、改变对所属领域技术人员来说都是显而易见了。
附图说明
图1显示了FGF1WT和FGF1△HS各种体外刺激的实验结果，其中，(A) 为FGF1WT、FGF1△HS和野生型FGF21 (FGF21WT)在3T3-L1脂肪细胞中对FGFR, FGFR底物2（FRS2）和MAPK途径(ERK1/2) 的活化的免疫印迹图； (B) 为 FGF1WT、FGF1△HS和野生型 FGF19 (FGF19WT) 在大鼠肝细胞瘤细胞系H4 E中对FGFR, FGFR底物2（FRS2）和MAPK途径(ERK1/2) 的活化的免疫印迹图； (C) 为FGF1WT、FGF1△HS和FGF21WT刺激3T3-L1脂肪细胞所产生的细胞糖摄入情况；(D) 为FGF1WT、FGF1△HS和FGF19WT刺激大鼠肝细胞瘤细胞系H4E产生的细胞糖摄入情况；(E) 为SPR检测图谱；(F) 为HPLC-MALS检测图谱 (G) 显示了FGF1WT 和FGF1△HS为对NIH 3T3细胞的增殖效应，*p<0.05 vs PBS缓冲液（对照）；(H) 为FGF1WT和FGF1△HS在NIH 3T3细胞中对FGFR、FRS2和 MAPK途径 (ERK1/2) 的活化的免疫印迹图；(I) 为100 ng/ml FGF1WT和FGF1△HS在NIH 3T3细胞中对FGFR、FRS2和 MAPK途径 (ERK1/2) 的活化的免疫印迹图。
图2显示了FGF1WT和FGF1△HS对体内肝组织的促有丝分裂活性，其中，(A) 为用增殖细胞核抗原(PCNA) 和Ki67染色的鼠肝组织切片镜检图；(B) 为用FGF1WT和FGF1△HS长期给药的小鼠的肝组织蛋白表达的免疫印迹分析，*p<0.05 vs 对照 (用PBS给药)， #p<0.05 vsFGF1WT。
图3显示了FGF1WT和FGF1△HS对 db/db小鼠中血糖和胰岛素敏感性的影响，其中，(A) 为单次注射FGF1WT和FGF1△HS对 db/db小鼠中24小时血糖水平的影响，*p<0.05 vs db/ m; #p<0.05 vs db/db. (B) 显示了FGF1WT和FGF1△HS对 db/db小鼠中血糖降低的剂量依赖性影响； (C) 为单次注射FGF1WT和FGF1△HS后6小时进行的GTT的血糖水平，(D) 为单次注射FGF1WT和FGF1△HS后6小时进行的GTT的AUC，*p<0.05 vs db/m; #p<0.05 vs db/db；(E) 为单次注射FGF1WT和FGF1△HS后6小时进行的ITT的血糖水平，(F) 为单次注射FGF1WT和FGF1△HS后6小时进行的ITT的AUC，*p<0.05 vs db/m; #p<0.05 vs db/db；(G) 为用FGF1WT和FGF1△HS经28天给药的db/db小鼠的血糖水平，*p<0.05 vs db/m; #p<0.05 vs db/db。
图4显示了FGF1WT和FGF1△HS对db/db小鼠中肝脏脂和糖代谢的长期影响，其中，(A)为用H&E、Oil Red O和PAS染色的鼠肝组织切片镜检图；(B) 为用FGF1WT和FGF1△HS经28天给药的db/db小鼠的肝组织中的甘油三酯含量， (C) 为用FGF1WT和FGF1△HS经28天给药的db/ db小鼠的肝组织中的糖原含量， *p<0.05 vs db/m; #p<0.05 vs db/db；(D-G)显示了肝脏各蛋白质表达的免疫印迹分析，(H-J)显示了肝脏各mRNA表达的实时定量PCR分析，*p<0.05vs db/m; #p<0.05 vs db/db。
图5显示了FGF1WT和FGF1△HS对db/db小鼠中脂肪组织重塑和脂代谢的长期影响，其中， (A) 为用H&E染色的鼠白色脂肪组织(WAT)切片镜检图；(B) 为用H&E染色的鼠棕色脂肪组织(BAT)切片镜检图； (C-G) 显示了WAT各mRNA表达的实时定量PCR分析，*p<0.05 vsdb/m; #p<0.05 vs db/db。
图6显示了FGF1WT和FGF1△HS对db/db小鼠全身和肝脏炎症的长期影响，其中，(A)显示了用FGF1WT和FGF1△HS经28天给药的db/db小鼠血浆中的IL-6浓度， (B) 显示了用FGF1WT和FGF1△HS经28天给药的db/db小鼠血浆中的TNF-α浓度， *p<0.05 vs db/m; #p<0.05 vs db/db；(C) 为对CD68表达染色的肝组织镜检图； (D-G) 显示了肝脏各炎性因子的蛋白质表达的免疫印迹分析，(H-J) 显示了肝脏各炎性因子的mRNA表达的实时定量PCR分析，*p<0.05 vs db/m; #p<0.05 vs db/db。
具体实施方式
以下通过实施例进一步说明本发明的内容。如未特别指明，实施例中所用的技术手段为本领域技术人员所熟知的常规手段和市售的常用仪器、试剂，可参见《分子克隆实验指南（第3版）》（科学出版社）和CFDA的相关试验指引以及相应仪器和试剂的厂商说明书等参考。
实施例1 本发明的FGF1△HS及其体外活性研究
通过常规手段克隆、表达并纯化FGF1△HS。简而言之，将编码全长人野生型FGF1(简称为FGF1WT, 第1-155位)的cDNA片段克隆入pET30a表达载体中，然后用QuikChange XL定点突变试剂盒(Stratagene, La Jolla, CA)导入Lys127Asp、Lys128Gln和Lys133Val这三个突变。将带有突变体的表达载体转化入Escherichia coli BL21(DE3)中，于37 °C培养，当A600达到0.5时，加入1 mM IPTG继续培养4小时。菌体裂解后用依次阳离子交换层析柱和凝胶排阻层析柱(GE Healthcare, Piscataway, NJ)纯化，获得纯度>98%的FGF1△HS。其他对照蛋白也可以用相似方法制备。
通过SPR生物感应芯片(BIAcore 2000 system (GE Healthcare, Piscataway,NJ))检测，相比于FGF1WT，FGF1△HS的肝素结合亲和性得到了显著的丧失(参见图1E)。
通过HPLC-MALS分析，其中HPLC (Waters 1500 pump with 2498 UV detectorand 2707 autosampler) 偶联MALS (Wyatt miniDawn-Treos and Optilab rEX) ，来比较在肝素十糖的存在下， FGF1WT和FGF1△HS分别对FGFR1c二聚化的诱导能力。结果如图1F所示，肝素十糖导致了FGF1WT-FGFR1c二聚化复合物，其分子量通过MALS测得为89.54 kDa (理论二聚体值为89.48 kDa)；而FGF1△HS-FGFR1c复合物的保留时间明显延长，分子量仅为57.64 kDa。
在培养3T3L1脂肪细胞和大鼠肝细胞瘤细胞H4E时加入不同浓度的FGF1WT和FGF1△HS，发现相对于FGF1WT，FGF1△HS对FGFR的A-loop酪氨酸的磷酸化能力弱了两个数量级，而对FRS2α-RAS-MAPK途径的活化更接近于FGF19 和FGF21(图1A和1B)。尽管如此不同，但是在3T3L1和H4IIE细胞中，FGF1△HS仍旧保留了类似于FGF1WT的刺激葡萄糖摄取的效应(图1C和1D)
用NIH 3T3 成纤维细胞比较FGF1WT和FGF1△HS的促有丝分裂能力，结果如图1G所示，相比于FGF1WT，FGF1△HS的促有丝分裂能力得到了显著的丧失；另外如图1H和1I所示，FGF1△HS对FGFR的A-loop酪氨酸的磷酸化能力弱了至少1个数量级，并且减弱了诱导FRS2磷酸化和ERK活化的能力。
实施例2 本发明的FGF1△HS的体内活性研究
（1）正常小鼠
每天对正常的C57BL/6J小鼠给药FGF1WT和 FGF1△HS (0.5 mg/kg体重)，持续3个月，分别用PCNA和Ki67 (图2A) 免疫组化染色和Western blotting (图2B)对小鼠的肝脏分析增生情况。FGF1WT被观察到能导致小鼠增生，而FGF1△HS相对于PBS的对照组，没有出现增生。
（2）FGF1WT和FGF1△HS对db/db小鼠的血糖水平和胰岛素敏感性的影响
糖尿病模型(db/db)小鼠(C57BLKS/J-lepr db /lepr db ) 和其对照的表型正常的小鼠(db/m) 均购自南京大学模型动物研究中心。
在db/db小鼠中，快速注射FGF1WT和FGF1△HS (0.5 mg/kg 体重)都能显著降低血糖水平，该效果都基本能持续到注射后的24小时 (图3A)，而且，该效果都是剂量依赖性的(图3B)。
在糖耐测试(GTT)中，快速注射FGF1WT和FGF1△HS的db/db小鼠都能保持较低的血糖水平 (图3C 和3D)；而在胰岛素耐受测试(ITT)中, 注射FGF1WT和FGF1△HS的db/db小鼠都显示出了显著的胰岛素敏感性的改善 (图3E和3F)。
通过隔天注射db/db小鼠0.5 mg/kg的FGF1WT和FGF1△HS，持续4周，来观察长期给药的效果。结果如图3G所示，FGF1WT和FGF1△HS都能使db/db小鼠的血糖水平在4周恢复正常。
（3）FGF1WT和FGF1△HS对db/db小鼠的肝脏脂和糖代谢的长期影响
前述长期给药的小鼠及其对照小鼠中取肝脏组织用不同染料染色，结果如图4A所示，其中未给药的db/db小鼠有着显著的肝脂肪变性，而用FGF1WT和FGF1△HS长期给药的db/db小鼠都减轻了肝脂肪变性的程度。
长期给药的db/db小鼠的肝脏的甘油三酯水平也显著降低，接近了表型正常的db/ m小鼠(图4B)。肝组织中的脂积累的减少表明脂肪生成和脂肪储存的减少，相应地，在长期给药的db/db小鼠的肝组织中大量脂肪生成基因的表达蛋白及其mRNA水平均向db/m小鼠的靠拢(图4D-4J)。
小鼠的肝糖原水平通过PAS染色，结果如图4A所示，db/db小鼠相对于db/m小鼠提高了肝糖原水平，而用FGF1WT和FGF1△HS长期给药的db/db小鼠的肝糖原水平进一步增加了。定量检测也显示了这样的结果（图4C）。表明FGF1WT和FGF1△HS的降糖和提高胰岛素敏感性是通过增加肝糖原的合成和储存来实现的。
（4）FGF1WT和FGF1△HS对db/db小鼠的脂肪重塑和脂代谢的长期影响
肥胖症和2型糖尿病均油增多的脂肪储存和白色脂肪细胞(WAT)扩充的特征，而用FGF1WT和FGF1△HS长期给药的db/db小鼠都减少了白色和棕色脂肪细胞（BAT）的大小（图6A和6B）；相应的WAT中的大量脂肪生成基因的表达也向正常水平靠拢（图6C-6G）。
（5）FGF1WT和FGF1△HS对db/db小鼠的全身性炎症的长期影响
如图6A和6B所示，用FGF1WT和FGF1△HS长期给药的db/db小鼠都能降低血浆中炎性细胞因子IL-6和TNF-α的水平。db/db小鼠的肝脏中具有高的CD68+巨噬细胞浸润水平（图6C），并且CD68和炎性细胞因子IL-6和TNF-α的蛋白和RNA水平都被上调（图6D-6J）；而用FGF1WT和FGF1△HS长期给药的db/db小鼠的肝脏中的巨噬细胞浸润、CD68和炎性细胞因子的表达都显著降低（图6C-6J），几乎达到正常db/m小鼠的水平。
SEQUENCE LISTING
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Ala Met Asp Thr Asp Gly Leu Leu Tyr Gly Ser Gln Thr Pro Asn Glu
85 90 95
Glu Cys Leu Phe Leu Glu Arg Leu Glu Glu Asn His Tyr Asn Thr Tyr
100 105 110
Ile Ser Lys Lys His Ala Glu Lys Asn Trp Phe Val Gly Leu Lys Lys
115 120 125
Asn Gly Ser Cys Lys Arg Gly Pro Arg Thr His Tyr Gly Gln Lys Ala
130 135 140
Ile Leu Phe Leu Pro Leu Pro Val Ser Ser Asp
145 150 155
Claims (10)
1.FGF1突变体，其氨基酸序列在SEQ ID NO：1的基础上具有Lys127、Lys128和/或Lys133突变。
2.权利要求1所述的突变体，其具有Lys127Asp、Lys128Gln和/或Lys133Val突变。
3.编码权利要求1或2所述的突变体的多核苷酸。
4.载体或细胞，其包含权利要求3所述的多核苷酸。
5.药物组合物，其包括权利要求1或2所述的突变体以及药学上可接受的载体。
6.权利要求1或2所述的突变体在制备减少FGF1促细胞分裂和增殖副作用的药物中的应用。
7.权利要求6所述的应用，其中药物用于治疗糖尿病，尤其是2型糖尿病。
8.权利要求6所述的应用，其中药物用于降血糖。
9.权利要求6所述的应用，其中药物用于治疗或预防肥胖症。
10.权利要求6所述的应用，其中药物用于抗炎或者用于降低炎性细胞因子（如，IL-6和/或TNF-α）水平。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050227329A1 (en) * | 1985-09-12 | 2005-10-13 | Fiddes John C | Recombinant fibroblast growth factor analogs |
| CN101906162A (zh) * | 2009-06-04 | 2010-12-08 | 温州医学院 | 成纤维细胞生长因子-1及其突变体的聚乙二醇修饰产物 |
| CN103597076A (zh) * | 2011-03-01 | 2014-02-19 | 人体酶公司 | 成纤维细胞生长因子的热稳定变体 |
| CN104736557A (zh) * | 2012-06-07 | 2015-06-24 | 纽约大学 | 嵌合成纤维细胞生长因子21蛋白及其使用方法 |
| CN105828833A (zh) * | 2013-10-21 | 2016-08-03 | 萨克生物研究学院 | 突变的成纤维细胞生长因子（fgf）1及使用方法 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| GB8821795D0 (en) * | 1988-09-16 | 1988-10-19 | Erba Carlo Spa | New derivatives of human/bovine basic fibroplast growth factor |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050227329A1 (en) * | 1985-09-12 | 2005-10-13 | Fiddes John C | Recombinant fibroblast growth factor analogs |
| CN101906162A (zh) * | 2009-06-04 | 2010-12-08 | 温州医学院 | 成纤维细胞生长因子-1及其突变体的聚乙二醇修饰产物 |
| CN103597076A (zh) * | 2011-03-01 | 2014-02-19 | 人体酶公司 | 成纤维细胞生长因子的热稳定变体 |
| CN104736557A (zh) * | 2012-06-07 | 2015-06-24 | 纽约大学 | 嵌合成纤维细胞生长因子21蛋白及其使用方法 |
| CN105828833A (zh) * | 2013-10-21 | 2016-08-03 | 萨克生物研究学院 | 突变的成纤维细胞生长因子（fgf）1及使用方法 |
Non-Patent Citations (1)
| Title |
|---|
| MALGORZATA ZAKRZEWSKA等: "Increased Protein Stability of FGF1 Can Compensate for Its Reduced Affinity for Heparin", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
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