CN103041387B - 恶性b细胞淋巴瘤抗体药物的新用途 - Google Patents
恶性b细胞淋巴瘤抗体药物的新用途 Download PDFInfo
- Publication number
- CN103041387B CN103041387B CN201110306828.6A CN201110306828A CN103041387B CN 103041387 B CN103041387 B CN 103041387B CN 201110306828 A CN201110306828 A CN 201110306828A CN 103041387 B CN103041387 B CN 103041387B
- Authority
- CN
- China
- Prior art keywords
- antibody
- lymphoma
- leukemia
- hla
- light chain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000003814 drug Substances 0.000 title claims abstract description 15
- 230000003211 malignant effect Effects 0.000 title abstract description 8
- 208000003950 B-cell lymphoma Diseases 0.000 title abstract description 4
- 206010025323 Lymphomas Diseases 0.000 claims abstract description 36
- 208000032839 leukemia Diseases 0.000 claims abstract description 23
- 238000011282 treatment Methods 0.000 claims abstract description 23
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 4
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 claims description 3
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 claims description 2
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 claims description 2
- 208000031261 Acute myeloid leukaemia Diseases 0.000 claims description 2
- 208000036762 Acute promyelocytic leukaemia Diseases 0.000 claims description 2
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 claims description 2
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 claims description 2
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 claims description 2
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 23
- 206010028980 Neoplasm Diseases 0.000 description 12
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 12
- 230000000694 effects Effects 0.000 description 10
- 239000000427 antigen Substances 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 210000004881 tumor cell Anatomy 0.000 description 7
- 238000002512 chemotherapy Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- RTQWWZBSTRGEAV-PKHIMPSTSA-N 2-[[(2s)-2-[bis(carboxymethyl)amino]-3-[4-(methylcarbamoylamino)phenyl]propyl]-[2-[bis(carboxymethyl)amino]propyl]amino]acetic acid Chemical compound CNC(=O)NC1=CC=C(C[C@@H](CN(CC(C)N(CC(O)=O)CC(O)=O)CC(O)=O)N(CC(O)=O)CC(O)=O)C=C1 RTQWWZBSTRGEAV-PKHIMPSTSA-N 0.000 description 5
- 210000001185 bone marrow Anatomy 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 229960001001 ibritumomab tiuxetan Drugs 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 229960005267 tositumomab Drugs 0.000 description 5
- 150000001413 amino acids Chemical group 0.000 description 4
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 210000003719 b-lymphocyte Anatomy 0.000 description 3
- 230000027455 binding Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000004393 prognosis Methods 0.000 description 3
- 238000001959 radiotherapy Methods 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 2
- 206010000830 Acute leukaemia Diseases 0.000 description 2
- 108010058597 HLA-DR Antigens Proteins 0.000 description 2
- 102000006354 HLA-DR Antigens Human genes 0.000 description 2
- 108010036449 HLA-DR10 antigen Proteins 0.000 description 2
- 241000053227 Themus Species 0.000 description 2
- 238000010322 bone marrow transplantation Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000002607 hemopoietic effect Effects 0.000 description 2
- 208000021760 high fever Diseases 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 206010025482 malaise Diseases 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011363 radioimmunotherapy Methods 0.000 description 2
- 201000005404 rubella Diseases 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 241000321096 Adenoides Species 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102100038080 B-cell receptor CD22 Human genes 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 206010051779 Bone marrow toxicity Diseases 0.000 description 1
- 108010065524 CD52 Antigen Proteins 0.000 description 1
- 102000013135 CD52 Antigen Human genes 0.000 description 1
- 241000272190 Falco peregrinus Species 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 description 1
- 206010020880 Hypertrophy Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 1
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 210000002534 adenoid Anatomy 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 229950003145 apolizumab Drugs 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 231100000366 bone marrow toxicity Toxicity 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- VDQQXEISLMTGAB-UHFFFAOYSA-N chloramine T Chemical compound [Na+].CC1=CC=C(S(=O)(=O)[N-]Cl)C=C1 VDQQXEISLMTGAB-UHFFFAOYSA-N 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 208000024207 chronic leukemia Diseases 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 201000001421 hyperglycemia Diseases 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 231100001022 leukopenia Toxicity 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 239000012516 mab select resin Substances 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- FWZLYKYJQSQEPN-SKLAJPBESA-N peregrine Chemical compound OC1[C@H]2[C@@H]3C4([C@@H]5C6OC(C)=O)C(OC)CC[C@@]5(C)CN(CC)[C@H]4C6[C@@]2(OC)C[C@H](OC)[C@H]1C3 FWZLYKYJQSQEPN-SKLAJPBESA-N 0.000 description 1
- FWZLYKYJQSQEPN-UHFFFAOYSA-N peregrine Natural products OC1C2C3C4(C5C6OC(C)=O)C(OC)CCC5(C)CN(CC)C4C6C2(OC)CC(OC)C1C3 FWZLYKYJQSQEPN-UHFFFAOYSA-N 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 210000004180 plasmocyte Anatomy 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 229960001479 tosylchloramide sodium Drugs 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明涉及恶性B细胞淋巴瘤抗体的新用途，公开了一种抗HLA-DR10的淋巴瘤特异性人源化抗体用于制备治疗白血病药物的用途，所述抗HLA-DR10的淋巴瘤特异性人源化抗体包括重链及轻链，其中，重链含恒定区，重链可变区氨基酸序列为SEQIDNO:1，轻链含恒定区，轻链可变区氨基酸序列为SEQIDNO:2，重链及轻链通过二硫键相连。本发明进一步拓展了抗HLA-DR10的淋巴瘤特异性人源化抗体在治疗白血病上的新用途，为白血病患者带来了福音。
Description
技术领域
本发明涉及已知恶性B细胞淋巴瘤抗体的新用途。
背景技术
恶性淋巴瘤(ML)是源于淋巴结和淋巴结外淋巴组织的恶性肿瘤，ML可分为何杰金氏淋巴瘤(HD)和非何杰金氏淋巴瘤(NHL)两大类。在我国，恶性淋巴瘤新发病例中，NHL患者占85～90％。在NHL中，预后较好的低度恶性淋巴瘤在我国的发病率较低，约占NHL的5％；而预后较差的高度恶性淋巴瘤发病率较高，占NHL的30％左右。调查显示，随着时间的推移和社会经济的发展，恶性淋巴瘤呈现增长的趋势，其中HD相对稳定，而NHL的发病率和死亡率均在上升。非何杰金氏淋巴瘤的病因与发病机制尚未确定，目前认为与性别、年龄、免疫功能缺失、细菌或病毒感染、环境等多种因素有关。
临床上治疗NHL的方法有化学治疗、放射治疗、手术治疗、骨髓移植等。目前对NHL的治疗以化疗为主，但化疗的短期副作用很明显，而且对复发病人作用不肯定，一般化疗联合方案效果不佳；放疗存在的问题是应用范围较小，而手术对NHL仅起诊断作用，极易复发，需要结合化疗；骨髓移植方法则由于配型困难、排异反应、价格昂贵等原因难以开展。
随着细胞生物学、分子生物学及生物工程技术的发展，免疫疗法在最近几年逐渐成为癌症治疗的主要方法之一。它直接或间接利用人体免疫系统内的免疫细胞及抗体来对抗癌症或减少因癌症或其治疗所引起的副作用，而肿瘤特异性的单克隆抗体类药物是目前免疫治疗的主导方向。
目前在国际上研究的用以治疗NHL的单抗类药物主要是靶向于淋巴细胞表面的抗原，已发表的研究结果中有针对CD20、CD22、CD52、CD19、CD30及HLA-DR等几种抗原的抗体产品。
由National Cancer Insititute研制的人源化IgG1单抗Apolizumab(MoAb Hu1D10)可作用在正常及恶性B淋巴细胞表面的HLA-DR抗原β链上的多个位点上产生抗体依赖性的细胞毒杀作用，在针对复发性或弥漫型CLL或ALL的I期临床试验中，有11％的病人治疗后达到PR，67％的病人为SD，22％的病人为PD。所有的病人都出现了1-2度输注反应(58％乏力、50％恶心、50％高血糖、42％发热、42％风疹和33％头痛)。该药现正处在II期临床试验研究阶段。
由Peregrine Pharmaceuticals公司研制的鼠源型单抗Lym-1(Oncolym)是一种能够特异性识别HLA-DR10抗原的鼠源性IgG2a单抗，该抗原在80％恶性淋巴瘤B细胞表面都有表达。它与淋巴瘤细胞的结合能力明显高于与正常B淋巴细胞的结合能力。在对125I-Lym-1的放射性显像研究中证实，Lym-1对已知的淋巴瘤组织部位有高度的选择性结合，而在正常的组织上结合较少。
在对Lym-1进行的在I期临床试验中，10例病人中有3例出现缓解。IgM HAMA反应均呈阴性。主要的毒性反应表现为：有8例病人出现了在注射近终点时或注射完成后的发热，其中4例最高体温保持在38℃以下，另4例最高体温保持在38～40℃之间，该发热很容易用药控制，而且通常只持续2～4小时；有4例病人在注射后出现了风疹；在研究过程中，没有发现心脏、肝肾或肺部毒性反应；而且在血液学指数方面也没有明显的改变；大多数病人的外周血淋巴细胞没有降低，免疫球蛋白的水平也未发现有明显改变。
放射免疫治疗(Radioimmunotherapy，RIT)则是一种免疫疗法和放射疗法相结合的治疗手段，它是以能与肿瘤抗原结合的物质作为靶向载体，偶联放射性核素作为“弹头”(治疗剂)的肿瘤治疗药物注入体内与肿瘤细胞相关抗原特异结合所致的肿瘤局部比其他正常组织剂量高，达到对肿瘤杀伤而对正常组织损伤小的一种治疗方法，而放免疗法中最常用的靶向载体是肿瘤细胞相关抗原的单克隆抗体。
目前在美国获准上市销售的治疗肿瘤的单抗类放射免疫药物主要有：Zevalin和Bexxar，输液反应及骨髓毒性是其主要毒副反应。3度或4度白细胞减少和血小板减少较为常见，并常发生于治疗后5-7周，并在2-4周后恢复。约5-7％接受Zevalin和Bexxar治疗的患者因感染需要住院治疗。少部分接受Zevalin和Bexxar治疗的患者可出现骨髓增生异常综合症和急性白血病，但这些病人往往先前接受过烷化剂治疗。研究还发现，若患者的骨髓受淋巴瘤侵犯越严重，则更多的放射性同位素聚集骨髓，从而对正常骨髓的损伤更大。因此，在使用Zevalin或Bexxar治疗前，必须仔细评价患者骨髓情况。淋巴瘤骨髓侵犯超过25％或发现骨髓功能不良者不宜接受Zevalin或Bexxar治疗。
由美国Peregrine Pharmaceuticals公司研制的鼠源型单抗Lym-1(Oncolym)既可以引导放射性核素靶向到肿瘤局部杀死肿瘤细胞，还具有ADCC、CDC及诱导Raji细胞凋亡的作用杀伤肿瘤细胞。Lym-1经放射性核素131I标记后形成的131I-Lym-1与大多数恶性B细胞表面的HLA-DR10抗原结合后可以引导放射性核素131I靶向到肿瘤局部杀死肿瘤细胞。
中国专利ZL200410053872.0公开了将Lym-1的可变区基因和人IgG1的γ1和κ恒定区基因融合，得到的一个低免疫原性的人鼠嵌合单抗(chLym-1)其中人源性序列为65％，鼠源性序列占35％。研究表明chLym-1有着和Lym-1相似的亲和力，而且在摄取后的24-72小时内chLym-1在肿瘤上的滞留量与Lym-1近乎相同。此外，chLym-1同样具有诱导ADCC、CDC效应及诱导淋巴瘤细胞凋亡效应来直接杀死肿瘤细胞的能力。它诱导ADCC效应的水平是鼠源型Lym-1的两倍，而且即便在较低的抗体浓度的条件下也可诱导最大的ADCC效应。因此新一代的人鼠嵌合型单克隆抗体chLym-1相对于鼠源型Lym-1具有表达率高、性能稳定、肿瘤摄取率高、正常组织摄取率低、可增加效应器功能、不易起HAMA反应而降低免疫源性及自身杀伤肿瘤细胞能力优秀等优点，优化了放免治疗效果。但该药物目前的研究仅限于肿瘤治疗上的应用，对于其它疾病的应用还有待开发及拓展。
中国专利申请200810034400.9进一步揭示了chlym-1在治疗恶性淋巴瘤及自身免疫性疾病中的应用。
白血病是造血组织的恶性疾病，又称“血癌”。其特点是骨髓及其它造血组织中有大量无核细胞无限制地增生，并进入外周血液，将正常血细胞的内核明显吸附，该病居年轻人恶性疾病中的首位，原生性病毒可能是神经性负感组织增生，还有许多因素如食物的矿物放射性化、毒化(苯等)或药物变异、遗传素质等可能是致病的辅因子。根据白血病细胞不成熟的程度和白血病的自然病程，分为急性和慢性两大类。我国急性白血病比慢性白血病多见(约5.5∶1)，其中急粒细胞白血病最多(1.62/10万)，其次为急淋白血病(0.39/10万)、慢粒白血病(0.36/10万)，慢淋白血病少见。目前对于白血病主要治疗有下列几类化学治疗、放射治疗、标靶治疗、中药治疗。部分高危险性病人，需要进行骨髓移植。近10年来，随着分子生物学、生物遗传学的进展，使白血病预后得到极大的改观。本发明根据研制单抗的作用机理，进一步研究了chlym-1在治疗白血病上的应用。
发明内容
本发明对现有的抗HLA-DR10的淋巴瘤特异性人源化抗体chLym-1的应用进一步拓展，提供了其在治疗白血病上的用途。
本发明一方面公开了一种抗HLA-DR10的淋巴瘤特异性人源化抗体用于制备治疗白血病药物的用途，所述抗HLA-DR10的淋巴瘤特异性人源化抗体包括重链及轻链，其中，重链含恒定区，重链可变区氨基酸序列为SEQ ID NO：1，轻链含恒定区，轻链可变区氨基酸序列为SEQ ID NO：2，重链及轻链通过二硫键相连。
所述白血病可选自急性淋巴细胞白血病，早幼粒细胞白血病，急性粒细胞白血病和慢性淋巴细胞白血病。
在一优选例中，所述抗体重链恒定区为γ类型；而抗体轻链恒定区为κ类型。
在另一优选例中，所述抗体是由NS0细胞株产生。
较佳的，所述抗体的重链的恒定区为人抗体重链的恒定区，所述抗体的轻链的恒定区为人抗体轻链的恒定区。
进一步的，所述抗HLA-DR10的淋巴瘤特异性人源化抗体经放射性同位素标记。
较佳的，上述放射性同位素选自131I、188Re、90Y或67Cu，优选131I。
本发明的抗体可作为主要活性成分与常用药用辅料配伍制成药物组合物用以治疗白血病。活性成分的给药量是治疗有效量。例如每天约1微克/千克体重-约10毫克/千克体重。此外，本发明的抗体还可与其他治疗剂一起使用。
本发明进一步拓展了抗HLA-DR10的淋巴瘤特异性人源化抗体在治疗白血病上的新用途，为白血病患者带来了福音。
具体实施方式
下面结合实施例进一步阐述本发明。应理解，这些实施例仅用于说明本发明，而非限制本发明的范围。
实施例1 抗HLA-DR10的淋巴瘤特异性人源化抗体的制备
采用中国专利(专利号：200410053872.0)中记载的保藏号为CCTCC NO.200405的小鼠骨髓瘤细胞株NSO-clym，将该细胞株接种于含10％血清、不含L-谷氨酰胺的SFM培养基中置于37℃，5％CO2培养箱中培养，收集培养上清，将过滤后的无细胞上清上样到MabSelect填充的FineLINE柱，上样的浓度是30mg/ml。洗脱后，洗脱液立即用磷酸钾中和pH值到6.8-7.0。再用凝胶过滤检测，结果表明纯度在95％以上。将产品测序，结果为，抗体的重链可变区氨基酸序列为SEQ ID NO：1，轻链可变区氨基酸序列为SEQ ID NO：2，符合预期。采用中国专利200410053872.0实施例4记载的方式检测该抗体的亲和常数Ka＝3.36X108mol-1
实施例2 131I标记的抗HLA-DR10的淋巴瘤特异性人源化抗体的制备
采用氯胺-T碘化标记法，将131I标记到实施例1获得的抗HLA-DR10的淋巴瘤特异性人源化抗体上，制得产物的放射性比活度为11mCi/mg。采用实施例1的方式检测该单抗的亲和常数Ka＝3.40X108mol-1。
实施例3 抗HLA-DR10的淋巴瘤特异性人源化抗体治疗白血病的药效学研究
试验方法：
(一)细胞准备
本实验检测的细胞均为悬浮细胞，实验检测48小时前1∶3传代细胞，细胞培养于含10％胎牛血清(Hyclone)的RPMI1640(GIBCO)培养基中。1000rpm离心收集细胞，PBS洗涤2次。
试验细胞系：淋巴瘤细胞系Raji、白血病细胞系HL-60。
(二)封闭
封闭的目的是为了减少抗体的非特异性结合，封闭剂为1％BSA，PBS pH 7.5。封闭30分钟。
(三)抗体孵育
一抗孵育：一抗为实施例1制备的人源化抗体ChLym-1抗体，孵育浓度为1ug/ml，孵育时间60分钟。PBST洗涤3次。
二抗孵育：二抗为Jackson公司的羊抗人FC片段抗体，使用浓度1ug/ml，孵育时间30分钟。PBST洗涤3次。
免疫荧光法中的二抗是荧光抗体，因此抗体孵育的时候必须避光。此外，为保证结合质量和防止干燥，抗体孵育在湿盒中进行。
(四)流式检测
流式检测各细胞表面ChLym-1抗原的表达情况。本实验使用的流式细胞仪为Beckman FC500，检测中使用只加二抗而未加入一抗的细胞作为对照细胞，以此计算各组细胞中阳性细胞的比例。
试验结果：
淋巴瘤细胞系Raji 阳性率70％--80％ MOLT-4 阳性率40％-50％
白血病细胞系HL-60 阳性率 27％--35％。
从结果上看，淋巴瘤细胞系表达较高，白血病细胞系有少数也表达lym-1，此试验说明ChLym-1抗体有望作用与白血病细胞起到一定的治疗作用。
Claims (4)
1.一种抗HLA-DR10的淋巴瘤特异性人源化抗体用于制备治疗白血病药物的用途，所述抗HLA-DR10的淋巴瘤特异性人源化抗体包括重链及轻链，其中，重链含恒定区，重链可变区氨基酸序列为SEQ ID NO:1，轻链含恒定区，轻链可变区氨基酸序列为SEQ ID NO:2，重链及轻链通过二硫键相连；所述抗体重链恒定区为γ类型；而抗体轻链恒定区为κ类型；所述抗体的重链的恒定区为人抗体重链的恒定区，所述抗体的轻链的恒定区为人抗体轻链的恒定区；所述白血病选自急性淋巴细胞白血病，早幼粒细胞白血病,急性粒细胞白血病和慢性淋巴细胞白血病。
2.如权利要求1所述抗HLA-DR10的淋巴瘤特异性人源化抗体的用途，其特征在于，所述抗体是由NS0细胞株产生。
3.如权利要求1所述抗HLA-DR10的淋巴瘤特异性人源化抗体的用途，其特征在于，所述抗HLA-DR10的淋巴瘤特异性人源化抗体经放射性同位素标记。
4.如权利要求3所述抗HLA-DR10的淋巴瘤特异性人源化抗体的用途，其特征在于，所述放射性同位素选自131I、188Re、90Y或67Cu。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110306828.6A CN103041387B (zh) | 2011-10-11 | 2011-10-11 | 恶性b细胞淋巴瘤抗体药物的新用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110306828.6A CN103041387B (zh) | 2011-10-11 | 2011-10-11 | 恶性b细胞淋巴瘤抗体药物的新用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103041387A CN103041387A (zh) | 2013-04-17 |
| CN103041387B true CN103041387B (zh) | 2014-12-10 |
Family
ID=48054396
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110306828.6A Active CN103041387B (zh) | 2011-10-11 | 2011-10-11 | 恶性b细胞淋巴瘤抗体药物的新用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103041387B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016197064A1 (en) * | 2015-06-04 | 2016-12-08 | Epstein Alan L | Lym-1 and lym-2 targeted car cell immunotherapy |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0316463A1 (en) * | 1987-11-13 | 1989-05-24 | Northwestern University | Murine hybridoma lym-1 and diagnostic antibody produced thereby |
| CN1737011A (zh) * | 2004-08-20 | 2006-02-22 | 上海美恩生物技术有限公司 | 抗hla－dr10的淋巴瘤特异性嵌合单抗 |
| CN101525385A (zh) * | 2008-03-07 | 2009-09-09 | 苏州工业园区晨健抗体组药物开发有限公司 | 恶性淋巴瘤、自身免疫性疾病抗体药物的筛选及其制备方法和用途 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6217871B1 (en) * | 1998-10-28 | 2001-04-17 | The Regents Of The University Of California | Antigenic epitopes with LYM-1 reactivity and uses thereof |
-
2011
- 2011-10-11 CN CN201110306828.6A patent/CN103041387B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0316463A1 (en) * | 1987-11-13 | 1989-05-24 | Northwestern University | Murine hybridoma lym-1 and diagnostic antibody produced thereby |
| CN1737011A (zh) * | 2004-08-20 | 2006-02-22 | 上海美恩生物技术有限公司 | 抗hla－dr10的淋巴瘤特异性嵌合单抗 |
| CN101525385A (zh) * | 2008-03-07 | 2009-09-09 | 苏州工业园区晨健抗体组药物开发有限公司 | 恶性淋巴瘤、自身免疫性疾病抗体药物的筛选及其制备方法和用途 |
Non-Patent Citations (5)
| Title |
|---|
| Gerald L. DeNardo et al..Nanomolecular HLA-DR10 Antibody Mimics: A Potent System for Molecular Targeted Therapy and Imaging.《CANCER BIOTHERAPY & RADIOPHARMACEUTICALS》.2008,第23卷(第6期),第783-795页. * |
| Gerald L. DeNardo et al..Nanomolecular HLA-DR10 Antibody Mimics: A Potent System for Molecular Targeted Therapy and Imaging.《CANCER BIOTHERAPY & * |
| RADIOPHARMACEUTICALS》.2008,第23卷(第6期),第783-795页. * |
| Rod Balhorn et al..Selective High-Affinity Ligand Antibody Mimics for Cancer Diagnosis and Therapy: Initial Application to Lymphoma/Leukemia.《Clinical Cancer Research》.2007,第13卷(第18 suppl期),摘要. * |
| Selective High-Affinity Ligand Antibody Mimics for Cancer Diagnosis and Therapy: Initial Application to Lymphoma/Leukemia;Rod Balhorn et al.;《Clinical Cancer Research》;20070915;第13卷(第18 suppl期);摘要 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103041387A (zh) | 2013-04-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tobinai et al. | Feasibility and pharmacokinetic study of a chimeric anti-CD20 monoclonal antibody (IDEC-C2B8, rituximab) in relapsed B-cell lymphoma | |
| CN101282745B (zh) | 用cd37-特异性和cd20-特异性结合分子减少b-细胞 | |
| Dobrenkov et al. | GD2-targeted immunotherapy and radioimmunotherapy | |
| AU2010328347B2 (en) | Methods for enhancing anti-tumor antibody therapy | |
| RU2658438C2 (ru) | Химерное терапевтическое антитело нн1 против cd-37 | |
| JP7161938B2 (ja) | スーパー抗原媒介性癌免疫療法の能力を増強するための方法および組成物 | |
| JP2024023438A (ja) | 多重特異性結合タンパク質及びその改善 | |
| AU2012273182A1 (en) | Focused radiation for augmenting immune-based therapies against neoplasms | |
| WO2019094626A1 (en) | Bispecific antibody compositions and related methods for improved pretargeted radioimunotherapies | |
| CN103214578B (zh) | 一种新型的人源化抗cd22抗体 | |
| JP2020529469A5 (zh) | ||
| JP2022090047A (ja) | 血液悪性疾患を処置するための低用量抗体に基づく方法 | |
| CN110312525A (zh) | T细胞重定向多功能抗体与免疫检查点调节剂的组合及其用途 | |
| WO2010145895A1 (en) | Compositions containing antibodies for treating cd5+ hla-dr+ b or t cell related diseases | |
| WO2022216965A1 (en) | Radioimmunotherapy directed to ccr8 for depletion of tumor infiltrating regulatory t cells | |
| Heijnen et al. | Generation of HER-2/neu-specific cytotoxic neutrophils in vivo: efficient arming of neutrophils by combined administration of granulocyte colony-stimulating factor and Fcgamma receptor I bispecific antibodies. | |
| KR20030086992A (ko) | 종양질환 치료를 위한 ｃｄ23 길항제의 용도 | |
| Coiffier | Monoclonal antibodies combined to chemotherapy for the treatment of patients with lymphoma | |
| Lindén et al. | A novel platform for radioimmunotherapy: extracorporeal depletion of biotinylated and 90Y-labeled rituximab in patients with refractory B-cell lymphoma | |
| CN103041387B (zh) | 恶性b细胞淋巴瘤抗体药物的新用途 | |
| AU686609B2 (en) | Compositons comprising IgG3 antibodies ***** Do not seal confirmation letter to come ******* | |
| KR20230028386A (ko) | 인간 질환의 치료를 위한 cd38 항체 | |
| Liu et al. | The potential for immunoconjugates in lymphoma therapy | |
| US5169775A (en) | Monoclonal antibodies against lymphoma-associated antigens, hybrid cell lines producing these antibodies | |
| CN101525385B (zh) | 恶性淋巴瘤、自身免疫性疾病抗体药物的筛选及其制备方法和用途 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190904 Address after: 518118 Unit 1505, Block A, Innovation Plaza, 2007 Pingshan Avenue, Pingshan Street, Pingshan District, Shenzhen City, Guangdong Province Patentee after: Yourui Biomedical Technology (Shenzhen) Co.,Ltd. Address before: 215125 Room 111, A3 Building, 218 Xinghu Street, Suzhou Industrial Park, Jiangsu Province Patentee before: Suzhou Industrial Park Chenjian Antibody Group Pharmaceutical Development Co.,Ltd. |
|
| PP01 | Preservation of patent right |
Effective date of registration: 20230522 Granted publication date: 20141210 |
|
| PP01 | Preservation of patent right | ||
| PD01 | Discharge of preservation of patent |
Date of cancellation: 20230815 Granted publication date: 20141210 |
|
| PD01 | Discharge of preservation of patent | ||
| CP03 | Change of name, title or address |
Address after: 518118 unit 1505, block a, innovation Plaza, No. 2007, Pingshan street, Pingshan District, Shenzhen, Guangdong Patentee after: Youjian Biopharmaceutical Technology (Shenzhen) Co.,Ltd. Country or region after: China Address before: 518118 unit 1505, block a, innovation Plaza, No. 2007, Pingshan street, Pingshan District, Shenzhen, Guangdong Patentee before: Yourui Biomedical Technology (Shenzhen) Co.,Ltd. Country or region before: China |
|
| CP03 | Change of name, title or address |