专利名称：预防或治疗肿瘤恶性变或肿瘤转移的组合物及方法
技术领域：
本发明涉及一种预防或治疗肿瘤恶性变或肿瘤转移的药物组合物及其方法，尤其是OPG在这种预防或治疗肿瘤恶性变或肿瘤转移的药物中的应用。
肿瘤转移是指恶性肿瘤细胞借助血道、淋巴道等途径，在非肿瘤原发部位的器官内形成继发瘤。目前发现的抗肿瘤转移剂，如维甲酰胺，能显著抑制大鼠前列腺癌细胞的生长和转移；常用的组合物(紫杉醇、顺铂、他莫昔等)对肿瘤细胞具有基本的作用，事实上对正常细胞具有很大的副作用。
恶性变和转移是恶性肿瘤的重要特征。癌转移使局部病变扩散成全身多灶性分布的疾病。在临床诊断出原发肿瘤时，一半以上的患者已经产生远隔部位的癌转移，以至现有的技术、放疗和化疗手段常常在治疗癌症时，无能为力，导致患者的死亡。因此防止肿瘤的恶性变以及肿瘤的转移，是延长肿瘤患者的生命，提高手术成功率的重要途径。
OPG是一种分泌型糖蛋白和缺乏跨膜结构域的TNF受体(TNFR)家族成员，OPG mRNA具有广泛的组织分布，在肝、心、肺、肾、胃、小肠、皮肤、脑、脊髓和骨骼中的表达水平较高。OPG是一种诱饵受体，其作用是结合或中和可溶性细胞核因子-κB受体活化因子配体[(RANKL)，也称破骨细胞分化因子、OPG配体(OPGL)和TNF相关活化诱导细胞因子(TRANCE)]及与基质细胞/成骨细胞结合的RANKL，从而阻断RANKL与破骨细胞系细胞表面的细胞核因子-κB受体活化因子[(RANK)，也称破骨细胞分化和活化受体]结合。RANKL配体一旦与RANK受体结合就会刺激前体破骨细胞分化，及活化成熟的破骨细胞，引起骨吸收。体外实验证实，OPG抑制前体破骨细胞分化和成熟破骨细胞形成骨吸收陷窝，并诱导破骨细胞凋亡。此外，OPG还可抵抗促骨吸收因子(PTH、PGE2、1，25(OH)2D3、糖皮质激素和RANKL等)引起的骨吸收。在正常生理条件下，OPG控制骨骼的骨质疏松和骨硬化两种极端表型。目前为止，所有关于OPG的功能性和病理性研究都集中在对骨的研究上，对于OPG在直接预防和治疗除了骨以外其他部位的肿瘤恶性变及肿瘤转移的应用尚未见到任何相关报道。
本发明的机理在于，应用受体与配体的理论，寻找可以直接抑制具有RANK受体的肿瘤的发生恶变及转移的多肽药物。
多肽生长因子与细胞因子是分泌因子，它们通过与不同的表面结合受体进行特异性结合，从而表明细胞生长、分化、代谢等变化的不同情况。作为一类蛋白，受体RANK在其结构和信号传导模式方面有所不同。它们的特征在于具有与配体RANKL结合有关的细胞外结构域和传递适当细胞内信号的胞质结构域。受体表达方式确定那种细胞会应答一种特定的配体，而特定受体的结构确定由配体结合并诱导的细胞应答。
正常细胞表面不表达特异性受体RANK，只有发生非典型性增生时，细胞表面产生少量的受体RANK。如果继续增殖，形成肿瘤细胞，这时产生大量的受体RANK。这种肿瘤细胞表面的受体RANK一旦与相应的配体RANKL结合后，随即发生肿瘤恶性变。OPG恰好能与该配体RANKL(此时可称为OPGL)优先结合，及时阻断肿瘤细胞的受体与配体的结合，阻止了该肿瘤细胞恶性变的发生。但是人体自身的OPG数量有限，只有当我们给患者注入一定剂量的OPG复合药物，才可以阻止大量肿瘤细胞恶性变的发生。
恶性变的肿瘤细胞沿着血管和淋巴道，进入在非肿瘤原发部位的器官组织内，当与该组织内大量存在的RANKL配体结合时，继续恶性变，这时产生肿瘤转移现象。我们如果预先给患者注入大量的OPG，就会抢先与配体RANKL结合，从而阻断了肿瘤细胞在该器官内的转移。
这就是OPG在预防和/或治疗肿瘤恶性变和肿瘤转移的药物中的应用。本发明提供的含有OPG及其OPG多肽的药物组合物对于淋巴结、骨、肝，肾、肾上腺、脑等肿瘤易转移的部位，都可以进行预防和治疗。
“OPG”或“OPG多肽”指序列表中的氨基酸序列的多肽及相关多肽。相关多肽包括等位基因变异体，剪接变异体，片段，衍生物，替代、缺失和插入的变异体，融合多肽和非人类同源体。
本发明还提供包括OPG融合蛋白和其变异体、片段和衍生物的药物组合物。本药物组合物包括不同的缓冲体系、PH和离子强度的稀释剂，还有一些常用的添加剂和填充剂，如抗氧化剂。该药物组合物可以采用本领域人员熟知的缓冲剂，也可以采用固体片剂的形式。总之，制剂中要包含OPG融合蛋白或其变异体、片段或衍生物，以及惰性成分。该药物组合物可以注射施用，也可以口服，或其他通用形式服用。
本发明优选液态药物进行皮下或肌肉注射的方式。
施用一定剂量的OPG融合多肽来预防或治疗肿瘤恶性变或肿瘤转移，有效量从0.1mg/kg到10mg/kg，优选从1mg/kg到10mg/kg，尤其优选3mg/kg到8mg/kg。
本发明药物对上皮细胞肿瘤恶变和腺癌具有显著疗效。尤其对腺癌的预防和治疗具有极好的疗效。
应用OPG及其相关多肽的药物预防和治疗肿瘤恶性变或肿瘤转移，不仅可以克服以往化学疗法的副作用，减轻病人的痛苦，而且提高了病人术后的治愈率。服用一定剂量OPG药物的病人，其肿瘤生长速度减慢，并且与没有服用该药的病人相比，用药病人癌细胞出现可测量生长量的时间比用药病人长2.5倍(5个月对2个月)。
本发明以下的实施例包含于但并不限制本发明的内容及范围。实施例1受体RANK在细胞增生不同阶段的数量检测1、第一组，取成年未孕的雌性正常小鼠(北京大学医学部提供)的乳腺细胞，检测细胞表面受体RANK含量，用RANK受体单克隆抗体免疫组化的方法进行DBA显色，无色(0)，表明不存在RANK受体。
2、第二组，取怀孕2周雌性小鼠的乳腺细胞，用同上1的方法DBA显色，阳性，淡黄色(+/-)，表明在乳腺细胞上有少量RANK受体。
3、将第二组怀孕小鼠在乳腺部位皮下注射乳腺癌细胞，3周后取小鼠的乳腺细胞用同上1的方法检测，强阳性，棕黄色(++)，表明此时有多量RANK受体在乳腺细胞上生成。
4、5周后取第二组小鼠的乳腺附近明显肿大的淋巴结，镜检有转移的肿瘤细胞出现，然后用同上1的方法检测，强阳性，深棕色(++++)，表明在该发生肿瘤转移的该部位已经生成大量的RANK受体。实施例2在小鼠模型中OPG对预防乳腺癌的作用取6-8周小鼠20只，将1*106个MDA-MB-435乳腺癌细胞静脉注射到小鼠的全身循环中，肿瘤接种后立即给小鼠静脉注射PBS缓冲液，然后给其中的10只小鼠注射含有有效剂量3mg/kg的OPG融合多肽药物混合物，每周3次，连续4周。通过对比，发现没有注射OPG药物的小鼠全部出现乳腺癌的症状，肿块直径1.5cm，而注射OPG药物的10只小鼠中只有4只有些微的肿瘤肿块，直径0.5cm，其余6只都没有明显肿瘤现象。实施例3在小鼠模型中OPG对治疗乳腺癌和防止肿瘤转移的作用对实施例2中的没有注射OPG的10只小鼠继续下列的实验。当注射乳腺癌细胞的4周后，10只小鼠的肿瘤已经有明显征兆，直径1.5cm，这时给其中的5只小鼠静脉注射含有有效剂量6mg/kg的OPG融合多肽药物混合物，每周4次，连续5周。通过对比，发现没有注射OPG药物的小鼠肿瘤明显继续扩大，而且在皮下有肿瘤转移发生，其中4只小鼠已经死去。而注射OPG药物的5只小鼠中，除了1只死去，其余4只小鼠肿瘤的生长有部分缓解，其中2只小鼠的肿块的生长完全缓解，并且没有肿瘤转移的发生。实施例4在小鼠模型中OPG对治疗前列腺癌中的作用按照实施例2的方法，取20只4-6周雄性裸鼠，皮下注射5*106个DU-145前列腺癌细胞诱导异体移植。在注射癌细胞的2周后，给其中10只小鼠静脉注射含有有效剂量3mg/kg的OPG多肽药物混合物，每周3次，连续5周。利用RT-PCR技术对血液中微转移癌细胞的检测，对两组小鼠的肿瘤细胞数量进行对比，发现注射OPG药物的一组小鼠中，肿瘤细胞的数量低于没有药物注射的一组。实施例5在小鼠模型中OPG对治疗肝癌的作用取20只4-6周小鼠，皮下注射1*106个人的SMMC-7721肝癌细胞(来自于中国科学院细胞库)，注射两周后，同样分为两组，每组10只，作为对照。一组小鼠注射含有有效剂量3mg/kg的OPG多肽药物混合物，每周3次，连续4周。利用RT-PCR技术对血液中微转移癌细胞的检测，对两组小鼠的肿瘤细胞数量进行对比，发现注射OPG药物的一组小鼠中，肿瘤细胞的数量低于没有药物注射的一组。实施例6在小鼠模型中OPG对治疗淋巴结转移的作用取成年雌性小鼠20只，将1*106个MDA-MB-435乳腺癌细胞静脉注射到小鼠的全身循环中，4天后，给其中一组小鼠注射含有有效剂量3mg/kg的OPG多肽药物混合物，每周3次，连续3周。另外一组不作任何处理。全部处死及自然死亡动物均经详细解剖学检查，测量肿瘤体积及侵袭和转移情况，观察乳腺附近的淋巴结有无转移。肿瘤组织及淋巴结经10％中性福尔马林固定、石蜡切片、HE染色。部分切片同时进行CEA(ABC法)免疫组织化学染色、光学显微镜观察。实验发现，没有注射OPG药物的小鼠，少数肿瘤细胞团的转移见于边缘窦，明显时侵入淋巴结实质，呈片块状。严重时几乎破坏全部淋巴结结构，发展成转移瘤结节。注射OPG药物的小鼠只有部分呈现轻微的淋巴结转移现象，症状并不明显。实施例7在小鼠模型中OPG对治疗多种肿瘤疾病的作用按照实施例2和实施例3的方法及步骤，给小鼠在相关的部位注射相关的肿瘤细胞，大肠癌，甲状腺癌，骨癌，肝癌，脑癌、肾上腺癌、肾癌、胃癌、大肠癌、肺癌等细胞，以及转移到淋巴结、肝、肾、骨、脑等部位的肿瘤细胞，然后采用同样的方法分组检测，发现注射OPG药物的一组小鼠中，肿瘤细胞的数量低于没有药物注射的一组，而且可以预防和治疗相关的肿瘤细胞转移。
序列表本发明中OPG的序列表采用AMGEN INC.在PCT/US00/22806申请进入中国时的文本。<210>1<211>232<212>蛋白质<213>人<400>1Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala1 5 10 15Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro20 25 30LyS Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val35 40 45Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val50 55 60Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln65 70 75 80Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln85 90 95Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala100 105 110Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro115 120 125Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr130 135 140Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser145 150 155 160Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr165 170 175Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr180 185 190Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe195 200 205Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys210 215 220Set Leu Set Leu Ser Pro Gly Lys225 230<210>2<211>401<212>蛋白质<213>人<400>2Met Asn Lys Trp Leu Cys Cys Ala Leu Leu Val Leu Leu Asp Ile Ile1 5 10 15Glu Trp Thr Thr Gln Glu Thr Leu Pro Pro Lys Tyr Leu His Tyr Asp20 25 30Pro Glu Thr Gly His Gln Leu Leu Cys Asp Lys Cys Ala Pro Gly Thr35 40 45Tyr Leu Lys Gln His Cys Thr Val Arg Arg Lys Thr Leu Cys Val Pro50 55 60Cys Pro Asp His Ser Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys65 70 75 80Val Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Ser Val LyS Gln Glu85 90 95Cys Asn Arg Thr His Asn Arg Val Cys Glu Cys Glu Glu Gly Arg Tyr100 105 110Leu Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Ser115 120 125Gly Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Lys130 135 140Cys Pro Asp Gly Phe Phe Ser Gly Glu Thr Ser Ser Lys Ala Pro Cys145 150 155 160Ile Lys His Thr Asn Cys Ser Thr Phe Gly Leu Leu Leu Ile Gln Lys165 170 175Gly Asn Ala Thr His Asp Asn Val Cys Ser Gly Asn Arg Glu Ala Thr180 185 190Gln Lys Cys Gly Ile Asp Ual Thr Leu Cys Glu Glu Ala Phe Phe Arg195 200 205Phe Ala Val Pro Thr Lys Ile Ile Pro Asn Trp Leu Ser Val Leu Val210 215 220Asp Ser Leu Pro Gly Thr Lys Val Asn Ala Glu Ser Val Glu Arg Ile225 230 235 240Lys Arg Arg His Ser Ser Gln Glu Gln Thr Phe Gln Leu Leu Lys Leu245 250 255Trp Lys His Gln Asn Arg Asp Gln Glu Met Val Lys Lys Ile Ile Gln260 265 270Asp Ile Asp Leu Cys Glu Ser Ser Val Gln Arg His Leu Gly His Ser275 280 285Asn Leu Thr Thr Glu Gln Leu Leu Ala Leu Met Glu Ser Leu Pro Gly290 295 300Lys Lys Ile Ser Pro Glu Glu Ile Glu Arg Thr Arg Lys Thr Cys Lys305 310 315 320Ser Ser Glu Gln Leu Leu Lys Leu Leu Ser Leu Trp Arg Ile Lys Asn325 330 335Gly Asp Gln Asp Thr Leu Lys Gly Leu Met Tyr Ala Leu Lys His Leu340 345 350Lys Thr Ser His Phe Pro Lys Thr Val Thr His Ser Leu Arg Lys Thr355 360 365Met Arg Phe Leu His Ser Phe Thr Met Tyr Arg Leu Tyr Gln Lys Leu370 375 380Phe Leu GIu Met Ile Gly Ash Gln Val Gln Ser Val Lys Ile Ser Cys385 390 395 400Leu<210>3<211>407<212>蛋白质<213>人<400>3Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His1 5 10 15Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His20 25 30Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr35 40 45Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro50 55 60Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His65 70 75 80Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyt Leu Glu Ile Glu Phe85 90 95Cys Leu Lys His Arg Ser Cys Pro Pro GIy Phe Gly Val Val Gln Ala100 105 110Gly Tyr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe115 120 125Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn130 135 140Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His145 150 155 160Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ala Ala Ala165 170 175Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala180 185 190Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro195 200 205Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val210 215 220Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val225 230 235 240Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln245 250 255Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln260 265 270Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala275 280 285Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro290 295 300Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr305 310 315 320Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser325 330 335Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr340 345 350Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr355 360 365Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe370 375 380Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys385 390 395 400Ser Leu Ser Leu Ser Pro Gly405<210>4<211>403<212>蛋白质<213>人<400>4Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His1 5 10 15Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His20 25 30Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr35 40 45Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro50 55 60Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His65 70 75 80Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe85 90 95Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala100 105 110Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe115 120 125Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn130 135 140Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His145 150 155 160Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Gly Ile165 170 175Asp Val Thr Ala Ala Ala Glu Pro Lys Ser Cys Asp Lys Thr His Thr180 185 190Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe195 200 205Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro210 215 220Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val225 230 235 240Lys Phe Ash Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr245 250 255Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val260 265 270Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys275 280 285Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser290 295 300Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro305 310 315 320Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val325 330 335Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly340 345 350Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp355 360 365Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thy Val Asp Lys Ser Arg Trp370 375 380Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His G1u Ala Leu His385 390 395 400Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly405 410<210>5<211>400<212>蛋白质<213>人<400>5<400>5Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His1 5 10 15Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His20 25 30Cys Thr Ala Lys Trp LyS Thr Val Cys Ala Pro Cys Pro Asp His Tyr35 40 45Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro50 55 60Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His65 70 75 80Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe85 90 95Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala100 105 110Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe115 120 125Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn130 125 140Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His145 150 155 160Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Val Asp Lys165 170 175Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro180 185 190Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser195 200 205Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp210 215 220Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Ual His Asn225 230 235 240Ala Lys Thr Lys Pro Arg GlU Glu Gln Tyr Asn Ser Thr Tyr Arg Val245 250 255Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu260 265 270Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys275 280 285Thr Ile Ser Lys Ala hys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr290 295 300Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr305 310 315 320Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu325 330 335Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu340 345 350Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys355 360 365Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu370 375 380Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly385 390 395 400<210>6<211>406<212>蛋白质<213>人<400>6Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His1 5 10 15Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His20 25 30Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr35 40 45Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro50 55 60Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His65 70 75 80Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe85 90 95Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala100 105 110Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe115 120 125Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn130 135 140Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His145 150 155 160Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Gly Ile165 170 175Asp Val Thr Val Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro180 185 190Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys195 200 205Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val210 215 220Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp225 230 235 240Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr245 250 255Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp260 265 270Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu275 280 285Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg290 295 300Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys305 310 315 320Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp325 330 335Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys340 345 350Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser355 360 365Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser370 375 380Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser385 390 395 400Leu Ser Leu Ser Pro Gly405<210>7<211>404<212>蛋白质<213>人<400>7Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His1 5 10 15Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His20 25 30Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr35 40 45Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro50 55 60Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His65 70 75 80Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe85 90 95Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala100 105 110Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe115 120 125Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn130 135 140Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His145 150 155 160Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ser Gly Gly165 170 175Gly Gly Gly Gly Gly Gly Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu180 185 190Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr195 200 205Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val210 215 220Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val225 230 235 240Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser245 250 255Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu260 265 270Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala275 280 285Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro290 295 300Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln305 310 315 320Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala325 330 335Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr340 345 350Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu355360 365Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser370 375 380Val Met His Glu Ala Leu His Asn His Tyr Thr Gln LVs Ser Leu Ser385 390 395 400Leu Ser Pro Gly<210>8<211>401<212>蛋白质<213>人<400>8Met Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu1 5 10 15Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu20 25 30Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser35 40 45His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu50 55 60Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr65 70 75 80Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn85 90 95Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro100 105 110Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln115 120 125Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val130 135 140Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val145 150 155 160Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro165 170 175Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr180 185 190Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val195 200 205Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu210 215 220Ser Pro Gly Lys Glu Thr Phe Pro Pro Lys Tyr Leu His Tyr Asp Glu225 230 235 240Glu Thr Ser His Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr245 250 255Leu Lys Gln His Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys260 265 270Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu275 280 285Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys290 295 300Asn Arg Thr His Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu305 310 315 320Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly325 330 335Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys340 345 350Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg355 360 365Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly370 375 380Asn Ala Thr His Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln385 390 395 400Lys
权利要求
1.一种预防或治疗哺乳动物肿瘤恶性变或肿瘤转移的方法，它包含施用有效剂量的OPG多肽。
2.权利要求1的方法，其中肿瘤发生部位包括乳腺、前列腺、肾上腺、甲状腺、胃、大肠、肺等腺体、器官和组织上，以及转移到淋巴结、肝、肾、骨、脑等部位。
3.权利要求1的方法，其中有效剂量的OPG多肽为0.1mg/kg-10mg/kg。
全文摘要
本发明公开了一种预防或治疗肿瘤恶性变或肿瘤转移的组合物及其方法，尤其是OPG在这种预防或治疗肿瘤恶性变或肿瘤转移的药物中的应用。具体的说，本发明公开了施用一种有效剂量的OPG融合蛋白和其变异体、片段和衍生物的药物组合物，可以预防或治疗肿瘤恶性变和肿瘤转移。
文档编号A61P35/00GK1471967SQ0212591
公开日2004年2月4日 申请日期2002年8月2日 优先权日2002年8月2日
发明者方健秋 申请人:方健秋