CN1169829C - 非t细胞结合肽及其用途 - Google Patents
非t细胞结合肽及其用途 Download PDFInfo
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- CN1169829C CN1169829C CNB021234124A CN02123412A CN1169829C CN 1169829 C CN1169829 C CN 1169829C CN B021234124 A CNB021234124 A CN B021234124A CN 02123412 A CN02123412 A CN 02123412A CN 1169829 C CN1169829 C CN 1169829C
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Abstract
本发明提供一种非T细胞结合肽及其类似物用于类风湿关节炎的治疗。这种多肽治疗剂可特异性抑制类风湿关节炎的异常免疫反应，针对发病的起始环节，从根本上控制本病的发展。
Description
本发明涉非T细胞结合肽的制备及其作为治疗类风湿关节炎和其它自身免疫病的药物的应用。
一、背景技术
类风湿关节炎(RA)是一种常见的以关节病变为主的慢性致残性自身免疫病。在中国的患病率为0.34％，全国有近500万患者，致残率高达93％。该病的发病过程是一种受抗原驱动的“激发—连锁免疫反应”的过程。感染因子及自身免疫反应介导的免疫损伤是类风湿关节炎发病及病情演变的基础。抗原多肽通过体内的抗原提呈细胞表达，并激活T淋巴细胞，导致细胞因子的释放、免疫球蛋白、趋化因子及自由基等炎性介质产生增多，进而引起血管炎，滑膜增生，软骨及骨破坏等类风湿关节炎的特征性病理变化。
目前，尚无从根本上控制类风湿关节炎的药物。临床上主要通过非甾体抗炎药如布洛芬及双氯芬酸等消炎止痛药暂时缓解症状。而慢作用抗风湿药如甲氨喋呤及来氟米特等则是通过抑制DNA合成广泛抑制免疫反应，进而抑制关节炎症。因此，这些药物在类风湿关节炎治疗中并非作用于发病的最初环节，难以针对性的控制病变，致使全身及关节病变不断进展，最终致残。而且，由于这些药物的广泛免疫抑制作用，骨髓抑制，肝功能异常等不良反应较多，致使很多患者不能接受这些药物治疗。
目前急需能够针对类风湿关节炎发病原因，通过抑制免疫反应的起始环节治疗本病的药物。为此，本发明人经过多年研究发现了可以用以治疗类风湿关节炎的非T细胞结合肽。研究证明，本申请中的多肽能够抑制HLA-DRβ1分子以及T细胞对抗原的识别和由此而致的自身免疫反应，从类风湿关节炎发病的中心环节遏止本病的发生和发展。
二、发明概述
本发明的目的之一是提供能够有效治疗类风湿关节炎的非T细胞结合肽。
本发明的另一目的是提供所述非T细胞结合肽在治疗类风湿关节炎中的用途。
最后，本发明提供了含有所述非T细胞结合肽的药物组合物。
本发明所用术语“非T细胞结合肽”指对SEQ ID NOS：1-7多肽中任意一个或多个氨基酸进行修饰后仍能够与HLA-DRβ1结合的多肽。用于修饰氨基酸的方法是本领域技术人员熟知的。
本发明主要是用丙氨酸(A)或甘氨酸(G)替换CII原形肽中一个或多个可与T细胞受体结合，并刺激T细胞增殖的氨基酸，保留与HLA-DRβ1结合的氨基酸，由此形成一类新的仅与HLA-DRβ1分子结合，而不被T细胞受体识别的CII多肽分子，即附件中所列的七种非T细胞结合肽(267A、268A、269A、270A、Mut269-270、Mut268-270、Mut267-270)，其序列及名称见序列表。
HLA-DRβ1、抗原多肽及T细胞受体三分子之间的相互识别和结合是类风湿关节炎异常免疫反应的中心环节。因此，本发明从导致类风湿关节炎的关键点——阻断T细胞对抗原的结合入手，利用非T细胞结合肽竞争性抑制致病抗原的作用及T细胞对抗原的识别，遏止类风湿关节炎患者体内的异常免疫反应，从而控制发病及病变的进展，达到从根本上治疗类风湿关节炎的目的。
已经证明，HLA-DRβ1中的第70-74位氨基酸含有一段共同序列QK/RRAA(即Gln-Lys/Arg-Arg-Ala-Ala，谷氨酰胺-赖氨酸/精氨酸-精氨酸-丙氨酸-丙氨酸)(1)，该段共同序列与HLA-DRβ1抗原结合槽的形成有关，是其结合抗原的功能氨基酸。本发明人和Wucherfennig等的大量研究证明该序列中带正电荷的第71位氨基酸(Lys或Arg)是抗原结合的关键部位(2-8)。
通过X线衍射技术对HLA-DRβ1-抗原二聚体结晶结构的研究发现，与类风湿关节炎相关的HLA-DRβ1(DR4/DR1)分子相结合的多种抗原肽在构型上均极其相似，其中包括变性II型胶原(CII)及热休克蛋白等(9-12)。从这些肽的立体结构(图1)可见，Phe263(P1)、Glu266(P4)和Gly271(P9)的侧链伸向左侧的HLA-DRβ1分子，全部或部分嵌入抗原结合槽内。而其它氨基酸的侧链则伸向一侧或与HLA-DRβ1相反的(T细胞受体)方向，刺激T细胞活化。由图2可见CII多肽的P1、P4、P9侧链嵌入HLA-DRβ1结合抗原的“口袋”中。带负电荷的P4(Glu)与HLA-DRβ1上带正电荷的第71位氨基酸(Lys71)相邻，形成高亲和力的极性结合。因此，Glu266可能是CII多肽结合DRβ1的关键氨基酸(13，14)。之后的研究进一步证明，CII主要靠Gln267、Gly268、Pro269和Lys270与T细胞受体结合，导致T细胞激活(12-14)。由此可见，CII多肽中主要与HLA-DRβ1结合的氨基酸为Phe263，Gly265，Glu266及Gly271、Glu272，而主要与T细胞受体结合的氨基酸为Gln267，Gly268，Pro269，Lys270(表1)。
表1.CII中与T细胞受体(TCR)和HLA-DRβ1结合的氨基酸
DRβ1 DRβ1 DRβ1 DRβ1 DRβ1
↑ ↑ ↑ ↑ ↑
F K G E Q G P K G E
263 264 265 266 267 268 269 270 271 272
↓ ↓ ↓ ↓
TCR TCR TCR TCR
*F＝Phe(苯丙氨酸)，K＝Lys(赖氨酸)，G＝Gly(甘氨酸)，E＝Glu(谷氨酸)，
Q＝Gln(谷氨酰胺)，P＝Pro(脯氨酸)，R＝Arg(精氨酸)，A＝Ala(丙氨酸)
基于以上的研究基础，本发明人将CII肽中与T细胞受体结合、刺激T细胞激活的氨基酸用丙氨酸(A)或甘氨酸(G)替换，保留与HLA-DRβ1结合的氨基酸，形成一种仅与DRβ1分子结合，而不被T细胞受体识别的非T细胞结合肽。这种非T细胞结合多肽可特异性结合HLA-DRβ1，竞争性抑制自身抗原与HLA-DRβ1的结合。从而抑制由HLA-DRβ1介导的自身免疫反应及由此引起的炎症介质释放等病理过程。
因此，在本发明的实施方案中，提供了非T细胞结合肽及其类似物，优选FKGEAGPKGE(SEQ ID NO：1)、FKGEQAPKGE(SEQ ID NO：2)、FKGEQGAKGE(SEQID NO：3)、FKGEQGPAGE(SEQ ID NO：4)、FKGEQGAAGE(SEQ ID NO：5)、FKGEQAGAGE(SEQ ID NO：6)及FKGEGAGAGE(SEQ ID NO：7)多肽。这些多肽可以与类风湿关节炎发病相关的HLA-DRβ1的特异序列QK/RRAA(即Gln-Lys/Arg-Arg-Ala-Ala)结合，从而使T细胞的激活受到抑制，达到治疗类风湿关节炎及其它T细胞介导的自身免疫病的目的。
在本发明的实施方案中还提供了含所述非T细胞结合肽或其类似物与可药用载体或佐剂的组合物。
这些可药用载体或佐剂包括乳糖、蔗糖、山梨醇、甘露醇、马铃薯淀粉、玉米淀粉或支链淀粉、纤维素衍生物、明胶、硬脂酸镁及硬脂酸钙等。可将所述组合物制成片剂、丸剂、胶囊剂、糖浆剂、粉剂、粒剂或溶液剂等。例如将所述药物组合用于口服给药时，该活性化合物可以与上述可药用载体或佐剂混合，然后压制成片剂。如果需要亦可包衣。
液体制剂还可以包括本领域技术人员熟知的其它赋形剂。该药物组合物中还可能包含药剂学允许的一些载体，如水、混悬剂和乳化剂等其它成分。
在本发明组合物中，非T细胞结合多肽或其类似物的含量通常为50-200μg/片，最佳为100μg/片。本领域技术人员根据患者的疾病情况、体重、年龄等可具体确定所施用的本发明非T细胞结合多肽的量。
本发明的非T细胞结合肽或其类似物可以通过多种途径(例如：口服、注射等)对关节炎患者给药。
附图说明
图1.已发表的与HLA-DRβ1结合的五个多肽(CII，HSP70，Clip，HA，HB)三维结构的相似性。5个多肽分别标以不同颜色重叠在一起，左右两个图从不同角度显示这些肽的结构。由图可见这些多肽的结构极其相似。左侧的第1(P1)，4(P2)，9(P9)位氨基酸侧链与HLA-DRβ1结合，全部或部分嵌入抗原结合槽内。而其它氨基酸的侧链多伸向一侧或与HLA-DRβ1相反的TCR方向、刺激T细胞的活化(Immunity 7：473-81，1997)。
图2.CII多肽与HLA-DRβ1结合的晶体结构。CII多肽的P1、P4和P9的侧链嵌入HLA-DRβ1的抗原结合“口袋”中。P4(Glu，带负电荷)与HLA-DRβ1上的Lys71(带正电荷)形成高亲和力的极性结合点(Immunity 7：473-81，1997)。
图3.非T细胞结合肽与CII原型多肽在T细胞激活作用的比较。与CII原形肽相比，Mut267-270，Mut268-270，Mut269-270，269A及270A多肽均无明显T细胞激活作用。
图4.非T细胞结合肽对T细胞激活的抑制作用。将非T细胞结合肽及原形肽与T细胞激活系统共同孵育时，非T细胞结合肽可明显抑制原形DRβ1结合肽的T细胞激活作用，而且，这种抑制作用随非T细胞结合肽浓度的增加而增强。
图5.CIA关节炎模型的滑膜病理特点。HE染色可见滑膜增厚和淋巴细胞浸润，部分关节可见血管翳形成；病理变化与关节肿胀率呈正相关。
图6.非T细胞结合肽(267A)对胶原性关节炎(CIA)的治疗作用。3个治疗组第8、10、12、14日的右足肿胀率和关节炎评分作均显著低于对照组(p＜0.05或0.01)；治疗后第8天和第16天，对照组CIA缓解率显著低于3个治疗组(p＜0.05)。
图7.非T细胞结合肽(267A)对胶原性关节炎(CIA)TNFα产生的抑制作用。100μg/ml剂量组外周血TNFα浓度显著低于对照组(p＜0.05)；其他两个治疗组与对照组相比无显著性差异。
以下通过实施例进一步说明本发明，但下列实施例不以任何方式限制本发明的范围。
实施例
实施例1：多肽的设计及合成
前述的研究工作(4，5，9-11)已经证明，CII多肽中267-270位氨基酸主要与T细胞受体结合，并激活T细胞。本发明的试验中首先利用Flechsler等(14)报道的固相法合成了一组七条含单一或数个氨基酸(CII267-270)替换的非T细胞结合肽(表2)。为提高这些多肽的吸收率、利用度及增强疗效，合成中将每条肽的氨基端与十四烷酸基因相连，以利多肽向细胞内转运。本研究中所用十肽均含有4个以上亲水残基，如赖氨酸(K)及谷氨酸(E)，易于溶解和吸收。这些肽不含易降解的甲硫氨酸(M)、色氨酸(W)、或易脱氨、脱羧基的天门冬氨酸(N)等。因此，肽的稳定性好，并且易于进入细胞内发挥其干扰T细胞识别的作用。
表2.非T细胞结合肽的设计
研究表明，上述7条非T细胞结合肽中与T细胞结合的氨基酸(267-270)分别由丙氨酸(A)或/和甘氨酸(G)替换，均具有抑制T细胞激活的作用，其中267A及Mut267-270多肽的显著效果已通过细胞系试验及关节炎动物模型证明(见后述)。
实施例2：本发明非T细胞结合肽对T细胞激活的抑制作用
抗原多肽通过抗原呈递细胞表达并激活T细胞，引起体内炎性介质的产生，进而引起关节炎的特征性病理变化。在实验中，通过测定T细胞增殖及白细胞介素(IL)-2水平检测非T细胞结合肽对T细胞激活的抑制作用，本研究中采用以下两套国际上认可的抗原呈递及T细胞激活系统：
抗原提呈细胞：L57.23细胞：识别HLA-DR1抗原(DR1转基因)
Priess细胞：识别HLA-DR1/4(EBV转染)
T细胞： 3.19细胞：HLA-DR1特异性T细胞株
3838细胞：HLA-DR4特异性T细胞株
其中L57.23细胞与3.19细胞为一套抗原提呈及T细胞激活系统，Priess细胞和3838细胞为另一套系统。通过比较上述七种非T细胞结合肽对抗原提呈及T细胞激活作用影响的差异，筛选出对T细胞激活有明显抑制作用的非T细胞结合肽。
在反应体系中首先加入抗原呈递细胞和不同的非T细胞结合肽(10μg/ml)及相应的T细胞，37℃培养48小时，取其上清，加入生长状态良好的IL-2依赖细胞(CTLL)中，应用四唑盐法(MTT)检测CTLL细胞增殖情况，从而了解非T细胞结合肽对T细胞激活的抑制效应。
本研究的结果显示，原型DRβ1结合肽在400、200、80、40、20μg/ml的浓度下均可刺激T细胞的增殖，上清中可检测到较高的IL-2浓度，而7条非T细胞结合肽对T细胞激活的刺激作用明显较弱，其上清中IL-2浓度显著低于原形肽(p＜0.01)(图3)。将非T细胞结合肽与原形肽及T细胞激活系统共同孵育时，非T细胞结合肽可明显抑制原形DRβ1结合肽的T细胞激活作用，而且，这种抑制作用随非T细胞结合肽浓度的增加而增强(图4)。进一步的试验证明，应用替换第267位氨基酸的非T细胞结合多肽治疗胶原诱导的关节炎大鼠，可以显著减轻关节肿胀和炎性反应。以上结果显示，原型CII263-272多肽含有II型胶原的抗原表位，替换该多肽中与T细胞结合的氨基酸，然后形成的非T细胞结合多肽可阻断T细胞受体对该多肽的识别，并可抑制T细胞的激活。这一系列的结果均提示非T细胞结合肽有可能成为干扰HLA-DR1/DR4介导的自身免疫反应的新方法。
实施例3：本发明非T细胞结合肽对实验性关节炎的抑制作用
在导致关节炎的免疫反应中，抗原呈递细胞是通过抗原诱导T细胞激活，并促使T细胞的增殖，引起动物体内的细胞因子(例如TNFα、IL-1)水平增高，进而导致类风湿关节炎的特征性病理变化。所以，通过检测实验动物体内的细胞因子的水平可了解非T细胞结合肽对关节炎的抑制作用。同时，通过病理切片也能从组织学的角度对药物疗效作出全面的评判。
在本发明的动物实验方案中，以牛CII诱导Wistar大鼠的胶原性关节炎(CIA)，然后用本发明中非T细胞结合肽对关节炎大鼠进行治疗，发现267A多肽可明显抑制大鼠关节炎，使关节肿胀程度减轻，病程缩短(见结果部分)。
(1)实验性关节炎动物模型的建立：
本发明中的实验性关节炎模型采用了国际公认的胶原性关节炎(CIA)(13，14)。实验动物为封闭群美国Wistar大鼠，本系大鼠为1907年由美国Wistar研究所培育而成，是我国引进最早，使用最广泛的品种之一。本实验用大鼠均在北京大学人民医院动物实验中心(洁净程度II级)饲养，大鼠被关闭在垫有木刨花的聚苯乙烯笼中，保持12小时白天/黑夜循环，使之可自由饮水和摄取食物。用于试验的大鼠均为8-10周龄的雄性大鼠，体重180±10g。实验中涉及动物的全部程序均按我国实验动物管理条理进行。
从Sigma公司购得牛II型胶原(批号：C1188)，用无水冰醋酸溶解配成8mg/ml的溶液，再用等体积的完全福氏佐剂(Difco，底特律，美国)乳化，在每只Wistar大鼠的右足底部注射100μl乳化物，注射剂量为400μl/只。本实验成功建立了Wistar大鼠的关节炎模型。
(2)对关节炎的评估：
在用CII诱导了Wistar大鼠的关节炎后约15天，全部大鼠均出现跖趾和趾间关节肿胀。用0-16的记分等级评估关节炎，对四只脚爪的每只按0-4记分，其中0＝没有关节肿胀，1＝1个关节肿胀，2＝2个关节肿胀，3＝3个关节肿胀，4＝全部关节肿胀，即整个脚爪肿胀。总的得分数由四只脚爪分别评分后累计得到。从免疫后2天开始，每天对大鼠进行一次检查，直至试验完成。
(3)非T细胞结合肽对Wistar大鼠的关节炎的治疗：
于Wistar大鼠出现关节炎的第4-7天，将出现关节炎的大鼠随机分为4组，其中3个治疗组，1个对照组，每组10只，每隔3天于大鼠尾根部皮下注射非T细胞结合肽267A，注射前用灭菌蒸馏水溶解，3个试验组使用的浓度分别为100μg/ml、500μg/ml、1000μg/ml，每只大鼠各100μl，对照组注射溶媒。每天用排水法测量右足体积变化，并用前述的关节炎评估方法对发生关节炎的关节数目作关节炎评分。治疗18天后处死动物，留取外周血，用TNFα和IL-2 ELISA试剂盒(晶美生物技术公司，深圳)检测其血清中的相应细胞因子的浓度；同时，取右足关节，固定，脱钙，切片做HE染色。
(4)应用SPSS软件包做统计分析。
(5)结果：本发明非T细胞结合肽对实验性关节炎的抑制作用
用牛II型胶原对40只雄性Wistar大鼠进行关节炎的诱导，结果全部大鼠于第14天至第17天时均出现不同程度的跖趾和趾间关节肿胀。在出现关节炎的第4-7天，将大鼠随机的分为4组，其中1个为对照组，其它3个为治疗组，每组10只。3个试验组使用的多肽267A的浓度分别为100μg/ml、500μg/ml、1000μg/ml，并用前述的关节炎评估方法对发生关节炎的关节数目评分。
结果显示，治疗前各组间右足肿胀率关节炎评分及外周血IL-2浓度无显著差异。所有出现肿胀的踝关节和跖趾关节的HE染色可见不同程度滑膜增厚和淋巴细胞浸润，部分关节可见血管翳形成(图5)，病理变化与关节肿胀呈正相关。3个治疗组于治疗后的2周内大鼠右足肿胀逐渐减轻，而对照组的右足肿胀无明显变化。
经过本发明非T细胞结合肽267A对关节炎大鼠进行治疗，用药后第4天三个治疗组大鼠的关节炎均开始明显减轻(图6，见下列表3)。3个治疗组关节炎的缓解率显著高于对照组(p＜0.05)，第8、10、12、14日的右足肿胀率和关节炎评分均显著低于对照组(p＜0.01)；3个治疗组间无显着性差异。100μg/ml剂量组外周血TNFα浓度显着低于对照组(p＜0.05)(图7)。这些结果证明，非T细胞结合肽可减轻关节炎大鼠的自身免疫性炎症，抑制胶原性关节炎的程度及病程，该多肽可能对类风湿关节炎有治疗作用。
表3.各组大鼠治疗右后足肿胀情况变化
| 编号 | 踝 第 第 第 第 第关 一 二 三 四 五节 趾 趾 趾 趾 趾 | 后足体积(ml) | 肿胀关节数 | 平均数 | |
| 对照组 | 1 | + + + + + + | 1.5 | 6 | 6.2 |
| 2 | - + ++ ++ ++ - | 1.4 | 7 | ||
| 3 | + ++ ++ + ++ - | 1.5 | 8 | ||
| 5 | + - ++ + + + | 1.3 | 6 | ||
| 6 | + - + + + ++ | 1.4 | 6 | ||
| 7 | + - + ++ + + | 1.3 | 6 | ||
| 8 | + ++ - + + + | 1.5 | 6 | ||
| 9 | + + + + + + | 1.4 | 7 | ||
| 10 | + + + - + - | 1.2 | 4 | ||
| 10mg组 | 1 | + - - + - - | 1.2 | 2 | 3.15* |
| 2 | - + + - + + | 1.5 | 4 | ||
| 3 | + + - - - + | 1.4 | 3 | ||
| 4 | - - - - + ++ | 1.3 | 3 | ||
| 5 | + - + - - ++ | 1.3 | 4 | ||
| 6 | + - + - - - | 1.3 | 2 | ||
| 7 | - + - - - - | 1.1 | 1 | ||
| 8 | + + ++ - + + | 1.3 | 6 | ||
| 9 | + - - - - - | 1.3 | 1 | ||
| 10 | + ++ + - - - | 1.2 | 4 | ||
| 50mg组 | 1 | + + + - + - | 1.4 | 4 | 3.25* |
| 2 | + + - ++ + + | 1.4 | 6 | ||
| 3 | + + + - + - | 1.4 | 4 | ||
| 4 | - + - + ++ ++ | 1.7 | 6 | ||
| 5 | + - - - + - | 1.3 | 2 | ||
| 6 | + - - - - - | 1.4 | 1 | ||
| 7 | + - - - - - | 1.1 | 1 | ||
| 8 | - - + - - - | 1.6 | 1 | ||
| 9 | + + - - - + | 1.6 | 3 | ||
| 10 | - - - + + + | 1.4 | 3 |
*与对照组相比，肿胀关节数明显减少(P＜0.01)
本发明人对其它本申请中非T细胞结合肽268A，269A，270A，Mut267-270及Mut267-270进行了上述T细胞激活和/或CIA模型的实验研究，均取得了与267A多肽类似的结果。
参考文献
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3.栗占国等：中国免疫学杂志2002，18：76-78。
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6.栗占国等：中华内科杂志2001，40：19-21。
7.栗占国等：中华医学杂志2001，81：111-113。
8.栗占国等：中华风湿病杂志2001，5：145-147。
9.Stern LJ，et al：Nature 368：215-221，1994。
10.Fremont DH，et al：Science 272：1001-1004，1996。
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12.Dessen A，et al：Immunity 7：473-81，1997。
13.Rosloniec EF，et al：J Exp Med.185：1113-1122，1997。
14.Anderson EC，et al：Proc.Natl.Acad.USA 95：7574-79，1998。
15.Flechsler I，etal：J Pept Scil(3)：141-200，1995。
序列表
<110>北京大学人民医院
<120>非T细胞结合肽及其用途
<140>
<141>2002-06-22
<160>7
<170>PatentIn version 3.1
<210>1
<211>10(氨基酸)
<212>肽
<213>人工序列
<223>合成
<400>1
Phe Lys Gly Glu Ala Gly Pro Lys Gly Glu
<210>2
<211>10
<212>肽
<213>人工序列
<223>合成
<400>2
Phe Lys Gly Glu Gln Ala Pro Lys Gly Glu
<210>3
<211>10
<212>肽
<213>人工序列
<223>合成
<400>3
Phe Lys Gly Glu Gln Gly Ala Lys Gly Glu
<210>4
<211>10
<212>肽
<213>人工序列
<400>4
Phe Lys Gly Glu Gln Gly Pro Ala Gly Glu
<210>5
<211>10
<212>肽
<213>人工序列
<400>5
Phe Lys Gly Glu Gln Gly Ala Ala Gly Glu
<210>6
<211>10
<212>肽
<213>人工序列
<400>6
Phe Lys Gly Glu Gln Ala Gly Ala Gly Glu
<210>7
<211>10
<212>肽
<213>人工序列
<400>7
Phe Lys Gly Glu Gly Ala Gly Ala Gly Glu
Claims (3)
1、选自下列任一序列的非T细胞结合肽：
FKGEAGPKGE(SEQ ID NO：1)
FKGEQAPKGE(SEQ ID NO：2)
FKGEQGAKGE(SEQ ID NO：3)
FKGEQGPAGE(SEQ ID NO：4)
FKGEQGAAGE(SEQ ID NO：5)
FKGEQAGAGE(SEQ ID NO：6)
FKGEGAGAGE(SEQ ID NO：7)。
2、含有权利要求1的非T细胞结合肽及可药用载体或佐剂的药物组合物。
3、权利要求1的非T细胞结合肽在制备治疗类风湿关节炎的药物中的应用。
Priority Applications (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021234124A CN1169829C (zh) | 2002-06-27 | 2002-06-27 | 非t细胞结合肽及其用途 |
| AU2003280443A AU2003280443A1 (en) | 2002-06-27 | 2003-06-26 | Non-t cell binding peptides and their uses |
| EP03739970A EP1541583A1 (en) | 2002-06-27 | 2003-06-26 | Non-t cell binding peptides and their uses |
| JP2004516425A JP2006508901A (ja) | 2002-06-27 | 2003-06-26 | Ｔ細胞非結合性ペプチド及びその用途 |
| PCT/CN2003/000496 WO2004003007A1 (fr) | 2002-06-27 | 2003-06-26 | Peptides se liant a des cellules non lymphocytaires, et utilisations correspondantes |
| US10/519,524 US7291600B2 (en) | 2002-06-27 | 2003-06-26 | Non-T cell binding peptides and their uses |
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| CNB021234124A CN1169829C (zh) | 2002-06-27 | 2002-06-27 | 非t细胞结合肽及其用途 |
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| CN1386754A CN1386754A (zh) | 2002-12-25 |
| CN1169829C true CN1169829C (zh) | 2004-10-06 |
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| US (1) | US7291600B2 (zh) |
| EP (1) | EP1541583A1 (zh) |
| JP (1) | JP2006508901A (zh) |
| CN (1) | CN1169829C (zh) |
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| CN103163302B (zh) * | 2011-12-10 | 2015-06-03 | 河北菲尼斯生物技术有限公司 | 一种采用定向交叉偶联技术制备的短肽抗体试剂盒 |
| WO2017155233A1 (ko) * | 2016-03-09 | 2017-09-14 | 주식회사 바이오펩 | 염증성 질환의 예방 또는 치료용 펩타이드 및 이의 용도 |
| CN110041404B (zh) * | 2019-04-08 | 2020-10-09 | 北京大学人民医院(北京大学第二临床医学院) | 瓜氨酸化抗原修饰肽及其应用 |
| CN117883547A (zh) * | 2021-07-30 | 2024-04-16 | 河北菲尼斯生物技术有限公司 | Se-dr亲和肽在制备治疗风湿疾病的药物中的用途 |
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| US6423315B1 (en) | 1993-03-03 | 2002-07-23 | The University Of Tennessee Research Corp. | Synthetic peptide for treatment of autoimmune arthritis |
| JPH08333390A (ja) | 1995-04-07 | 1996-12-17 | Hoechst Japan Ltd | ペプチド及びそれからなる自己免疫疾患治療剤 |
| DE69627855T2 (de) | 1995-06-13 | 2004-03-11 | Nippon Meat Packers, Inc. | Orales mittel gegen rheumatische arthritis und funktionelles nahrungsmittel |
| SE9700301D0 (sv) | 1997-01-31 | 1997-01-31 | Astra Ab | New compound |
| KR20010071020A (ko) | 1998-07-23 | 2001-07-28 | 에프.지.엠. 헤르만스 ; 이.에이치. 리링크 | 자가 면역 질환의 면역 요법에서 사용하기 위한 신규한펩티드 |
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2002
- 2002-06-27 CN CNB021234124A patent/CN1169829C/zh not_active Expired - Lifetime
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- 2003-06-26 AU AU2003280443A patent/AU2003280443A1/en not_active Abandoned
- 2003-06-26 JP JP2004516425A patent/JP2006508901A/ja active Pending
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| US20060166890A1 (en) | 2006-07-27 |
| EP1541583A1 (en) | 2005-06-15 |
| CN1386754A (zh) | 2002-12-25 |
| JP2006508901A (ja) | 2006-03-16 |
| US7291600B2 (en) | 2007-11-06 |
| AU2003280443A1 (en) | 2004-01-19 |
| WO2004003007A1 (fr) | 2004-01-08 |
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