醫(yī)學(xué)免疫學(xué)課件：雙特異性T細(xì)胞銜接器抗體

1、雙特異性T細(xì)胞銜接器抗體 Bispecific T-cell Engager (BiTE) Antibody1. 背景介紹1.1 腫瘤免疫治療的熱點(diǎn)1.2 雙特異性抗體的結(jié)構(gòu)1.3 無(wú)Fc段雙特異性抗體的發(fā)展1.4 雙特異性抗體的作用1.5 Fc段的作用1.6 雙特異性抗體的臨床應(yīng)用2. 作用機(jī)理2.1 BiTE的作用機(jī)理2.2 BiTE的抗原表位擴(kuò)展效應(yīng)3. 臨床研究3.1 Blinatumomab簡(jiǎn)介3.2 Blinatumomab的臨床試驗(yàn)結(jié)果4. 前景展望4.1 BiTE的優(yōu)點(diǎn)和缺點(diǎn)4.2 BiTE的改進(jìn)4.3 用于BiTE的腫瘤相關(guān)抗原4.4 雙特異性NK細(xì)胞銜接器(BiKE)4.5

2、 BiTE與免疫檢查點(diǎn)抑制劑的聯(lián)合4.6 溶瘤病毒表達(dá)BiTE4.7 自體T細(xì)胞表達(dá)BiTE腫瘤免疫治療的熱點(diǎn)單克隆抗體免疫檢驗(yàn)點(diǎn)單抗PD1/PDL1，CTLA4，CD47雙特異性抗體(BsAb)Triomab，BiTE，TandAb，DART過(guò)繼細(xì)胞療法腫瘤浸潤(rùn)淋巴細(xì)胞療法(TIL)T細(xì)胞受體療法(TCR)嵌合抗原受體修飾的T細(xì)胞療法(CAR)腫瘤疫苗樹(shù)突狀細(xì)胞(DC)疫苗蛋白/多肽/DNA疫苗定義：通過(guò)調(diào)動(dòng)宿主的免疫防御機(jī)制或給予某些生物活性物質(zhì)以取得或者增強(qiáng)腫瘤免疫效應(yīng)的治療方法。IgG抗體結(jié)構(gòu)雙特異性抗體的結(jié)構(gòu)Spiess C, Zhai Q, Carter P J. Alternat

3、ive molecular formats and therapeutic applications for bispecific antibodiesJ. Molecular immunology, 2015.目前已報(bào)道的雙特異性抗體結(jié)構(gòu)雙特異性抗體的結(jié)構(gòu)目前已進(jìn)入臨床試驗(yàn)的雙特異性抗體Kontermann R E, Brinkmann U. Bispecific antibodiesJ. Drug discovery today, 2015.無(wú)Fc段雙特異性抗體的發(fā)展Spiess C, Zhai Q, Carter P J. Alternative molecular formats an

4、d therapeutic applications for bispecific antibodiesJ. Molecular immunology, 2015.Geering B, Fussenegger M. Synthetic immunology: Modulating the human immune systemJ. Trends Biotechnol, 2014.無(wú)Fc段雙特異性抗體的發(fā)展（*代表已進(jìn)入臨床試驗(yàn)）已進(jìn)入臨床試驗(yàn)的無(wú)Fc段雙特異性抗體Diabody：VHa-VLb和VHb-VLascDiabody：VHa-VLb-VHb-VLaDART: VHa-VLb和VHb-

5、VLa 二硫鍵(VHa=VHb)TandAb: VHa-VLb-VHb-VLa 二聚化BiTE: VLa-VHa-VLb-VHbNanobody: VH雙特異性抗體的作用Kontermann R E, Brinkmann U. Bispecific antibodiesJ. Drug discovery today, 2015.重定向T細(xì)胞或NK細(xì)胞殺傷腫瘤細(xì)胞結(jié)合兩種細(xì)胞受體或配體，抑制信號(hào)轉(zhuǎn)導(dǎo)催化兩種蛋白相互作用雙特異性抗體的三種作用機(jī)制Fc段的作用Grandjenette C, Dicato M, Diederich M. Bispecific Antibodies: An Innova

6、tive Arsenal to Hunt, Grab and Destroy Cancer CellsJ. Current Pharmaceutical Biotechnology, 2015, 16(8): 670-683.具有Fc段的雙抗(如Triomab)發(fā)揮作用無(wú)Fc段的雙抗(如BiTE)發(fā)揮作用特點(diǎn)：容易提純，穩(wěn)定性高，半衰期長(zhǎng)（FcRn介導(dǎo)再回收）功能：結(jié)合輔佐細(xì)胞（NK，DC，M、粒），抗體依賴(lài)的細(xì)胞毒性（ADCC），補(bǔ)體依賴(lài)的細(xì)胞毒性（CDC）特點(diǎn)：穿透能力強(qiáng)，半衰期短（降解、腎清除）分子靶點(diǎn)結(jié)構(gòu)研發(fā)公司適應(yīng)癥臨床試驗(yàn)狀態(tài)注冊(cè)號(hào)Catumaxomab(Removab)EpCAM

7、CD3TrioMabFresenius Biotech(Trion)EpCAM陽(yáng)性腫瘤的惡性腹水2009EUEpCAM陽(yáng)性腫瘤的惡性腹水II/III完成NCT00836654胃癌、胃腺癌、食管胃連接部腺癌、腹膜轉(zhuǎn)移癌、卵巢癌、上皮卵巢癌、惡性腹水、輸卵管腫瘤、腹膜腫瘤II/III12項(xiàng)試驗(yàn)Ertumaxomab(Rexomun)Her2CD3TrioMabFresenius Biotech(Trion)Her2/Neu陽(yáng)性晚期固體瘤I/IINCT01569412Blinatumomab(Blincyto, MT103, AMG103)CD19CD3BiTEAmgen(Micromet)復(fù)發(fā)/難

8、治性費(fèi)城染色體陰性前體B細(xì)胞急性淋巴細(xì)胞白血病2014USA急性淋巴細(xì)胞白血病IINCT01466179非霍奇金淋巴瘤、急性淋巴細(xì)胞白血病、彌漫大B細(xì)胞淋巴瘤I/II/III16項(xiàng)試驗(yàn)MT110(AMG110)EpCAMCD3BiTEAmgen(Micromet)固體瘤I完成NCT00635596MT111(MEDI-565)CEACD3BiTEAmgen(Micromet)胃腸道腺癌I完成NCT01284231MT112(BAY2010112)PSMACD3BiTEBayer(Micromet)前列腺腫瘤INCT01723475AMG330CD33CD3BiTEAmgen(Micromet)

9、復(fù)發(fā)/難治性急性髓細(xì)胞白血病INCT02520427JNJ-64052781CD19CD3DARTB細(xì)胞慢性淋巴細(xì)胞白血病、彌漫大B細(xì)胞淋巴瘤、濾泡性淋巴瘤、套細(xì)胞淋巴瘤、急性淋巴細(xì)胞白血病INCT02454270MGD006CD123CD3DARTMacroGenicsand Sevier急性髓細(xì)胞白血病INCT02152956MGD007gpA33CD3DARTMacroGenicsand Sevier結(jié)直腸癌INCT02248805MGD009B7-H3CD3DARTMacroGenicsand Sevier間皮瘤、膀胱癌、黑色素瘤、頭頸部鱗狀細(xì)胞癌、卵巢癌、甲狀腺癌、乳腺癌、胰腺癌、前

10、列腺癌、結(jié)腸癌、軟組織肉瘤INCT02628535AFM13CD30CD16aTandAbAffimed霍奇金淋巴瘤I完成NCT01221571霍奇金淋巴瘤IINCT02321592AFM11CD19CD3TandAbAffimed復(fù)發(fā)/難治性B細(xì)胞非霍奇金淋巴瘤INCT02106091rM28HMW-MAACD28Tandem scFvUniversity HospitalTubingen惡性黑色素瘤I/II完成NCT00204594From by December 25, 2015目前已進(jìn)入臨床試驗(yàn)的用于連接效應(yīng)細(xì)胞與腫瘤細(xì)胞的雙特異性抗體BiTE的作用機(jī)理Oak E, Bartlett

11、N L. Blinatumomab for the treatment of B-cell lymphomaJ. Expert opinion on investigational drugs, 2015, 24(5): 715-724.Zimmerman Z, Maniar T, Nagorsen D. Unleashing the clinical power of T cells: CD19/CD3 bi-specific T cell engager (BiTE) antibody construct blinatumomab as a potential therapyJ. Inte

12、rnational immunology, 2014: dxu089.BiTE的結(jié)構(gòu)BiTE的作用機(jī)理BiTE與CD3和TAA結(jié)合，TCR-CD3復(fù)合物激活T細(xì)胞T細(xì)胞增殖，分泌細(xì)胞因子，CD25、CD69表達(dá)上調(diào)溶細(xì)胞突觸形成，釋放穿孔素和顆粒酶B穿孔素依賴(lài)Ca2+聚合形成跨膜通道，顆粒酶B進(jìn)入腫瘤細(xì)胞顆粒酶B激活Caspase通路，誘導(dǎo)腫瘤細(xì)胞凋亡ESK1-BiTEESK1：模擬TCR單克隆抗體，識(shí)別通過(guò)HLA-A*02:01呈遞的胞內(nèi)腫瘤蛋白WT1Dao T, Pankov D, Scott A, et al. Therapeutic bispecific T-cell engager

13、antibody targeting the intracellular oncoprotein WT1J. Nature biotechnology, 2015, 33(10): 1079-1086.BiTE的抗原表位擴(kuò)展效應(yīng)WT1-RMF和HER2/Neu陽(yáng)性卵巢癌患者的外周血單個(gè)核細(xì)胞(PMBC)給予ESK1-BiTE并接受自體腫瘤細(xì)胞刺激和再刺激后抗原表位特異性應(yīng)答B(yǎng)linatumomab簡(jiǎn)介商標(biāo)名Blincyto有效成分Blinatumomab生產(chǎn)商Amgen Inc.FDA批準(zhǔn)日期2014年12月適應(yīng)癥復(fù)發(fā)/難治性費(fèi)城染色體隱性前體B細(xì)胞急性淋巴細(xì)胞白血?。≧/R Ph- B-AL

14、L）半衰期2.1小時(shí)毒副作用細(xì)胞因子釋放綜合癥（CRS），神經(jīng)毒性等單抗種類(lèi)鼠源化BiTE結(jié)構(gòu)VLCD19-VHCD19-VHCD3-VLCD3靶點(diǎn)CD19CD3給藥方式靜脈費(fèi)用兩個(gè)療程共17.8萬(wàn)美元Przepiorka D, Ko C W, Deisseroth A, et al. FDA approval: blinatumomabJ. Clinical Cancer Research, 2015, 21(18): 4035-4039.Rogala B, Freyer C W, Ontiveros E P, et al. Blinatumomab: enlisting serial ki

15、ller T-cells in the war against hematologic malignanciesJ. Expert opinion on biological therapy, 2015, 15(6): 895-908.Blinatumomab的作用機(jī)理Blinatumomab的臨床試驗(yàn)結(jié)果注冊(cè)號(hào)研究人群樣本數(shù)平均年齡CR/CRhMRD陰性率無(wú)復(fù)發(fā)生存期總生存期NCT01466179II: R/R B-ALL18939(18-79)43%(36-51)82%(72-90)5.9 months(4.8-8.3)6.1 months(4.2-7.5)NCT00560794II: M

16、RD B-ALL2050(20-77)80%(56-94)NCT01207388II: MRD B-ALL11645(18-76)78%(69-85)NCT01209286II: R/R B-ALL3640(18-79)70%(52-84)69%(52-84)7.7 months(5.8-9.6)9.9 months(8.5-15.0)I: 兒童R/R B-ALL411832%77%8.3 months(3.0-16.0)5.7 months(3.3-9.7)I/II: 兒童R/R B-ALL399(2-16)31%(17%-48%)42%5.6 months(2.6-12.1)4.3 mon

17、ths(3.6-8.1)首次復(fù)發(fā)完全緩解(CR)：30-45%總生存期(OS)：5-9月原發(fā)難治/首次緩解小于12月/造血干細(xì)胞移植(HSCT)后復(fù)發(fā)/多線治療無(wú)效完全緩解(CR)：20-30%總生存期(OS)：3-6月Topp M S, Gkbuget N, Stein A S, et al. Safety and activity of blinatumomab for adult patients with relapsed or refractory B-precursor acute lymphoblastic leukaemia: a multicentre, single-arm

18、, phase 2 studyJ. The Lancet Oncology, 2015, 16(1): 57-66.Blinatumomab的I/II期臨床試驗(yàn)結(jié)果From by December 25, 2015BiTE的優(yōu)點(diǎn)和缺點(diǎn)優(yōu)點(diǎn)CD3TAA，重定向CD4+和CD8+T細(xì)胞（CD8為主，記憶T為主，Treg？）不依賴(lài)于TCR和MHCI呈遞，不需要共刺激腫瘤特異性半衰期短（隨時(shí)停止應(yīng)對(duì)突發(fā)）組織穿透能力強(qiáng)，調(diào)動(dòng)固體瘤內(nèi)TIL和Treg清除微小殘留疾病（MRD）療效好，相比單抗劑量低，低E:T比不易出現(xiàn)耐藥無(wú)基因操作、細(xì)胞移植風(fēng)險(xiǎn)，易獲得缺點(diǎn)半衰期短，難維持血清水平（持續(xù)靜脈給藥，結(jié)合PE

19、G或白蛋白）Laszlo G S, Gudgeon C J, Harrington K H, et al. T-cell ligands modulate the cytolytic activity of the CD33/CD3 BiTE antibody construct, AMG 330J. Blood cancer journal, 2015, 5(8): e340.Kontermann R E. Strategies for extended serum half-life of protein therapeuticsJ. Current opinion in biotechn

20、ology, 2011, 22(6): 868-876.Koristka S, Cartellieri M, Arndt C, et al. Tregs activated by bispecific antibodies: Killers or suppressors?J. OncoImmunology, 2015, 4(3): e994441.BiTE的改進(jìn)延長(zhǎng)半衰期改變配體腫瘤相關(guān)抗原多種CD3CD8？，CD16，CD28抗原結(jié)合位點(diǎn)多個(gè)與其他腫瘤療法結(jié)合Rituximab單抗免疫檢查點(diǎn)抑制劑體內(nèi)持續(xù)穩(wěn)定表達(dá)溶瘤病毒T細(xì)胞間充質(zhì)干細(xì)胞應(yīng)用于BiTE的腫瘤相關(guān)抗原靶點(diǎn)名稱(chēng)適應(yīng)癥臨床試驗(yàn)CD

21、19Blinatumomab/MT103/AMG103/MEDI-538急性淋巴細(xì)胞白血病非霍奇金淋巴瘤I/II/IIIEpCAMMT110固體瘤ICEAMT111/MEDI-565胃腸道腺癌IPSMAMT112/BAY2010112前列腺癌ICD33急性髓細(xì)胞白血病IEGFR結(jié)腸癌臨床前Her2臨床前EphA2bscEphA2CD3固體瘤臨床前MCSPMCSP-BiTE黑色素瘤臨床前ADAM17A300E前列腺癌臨床前PSCA前列腺癌臨床前17-1A臨床前NKG2D配體固體瘤、白血病臨床前CD133膠質(zhì)瘤臨床前WT1ESK1-BiTE固體瘤、白血病臨床前臨床和臨床前用于BiTE的腫瘤抗原靶點(diǎn)

22、Huehls A M, Coupet T A, Sentman C L. Bispecific T-cell engagers for cancer immunotherapyJ. Immunology and cell biology, 2014.雙特異性NK細(xì)胞銜接器(BiKE)CD16CD33BiKE對(duì)健康供者髓系抑制性細(xì)胞(MDSC)具有顯著殺傷作用骨髓增生異常綜合征(MDS)NK細(xì)胞 CD16 髓系抑制性細(xì)胞(MDSC)CD16CD33激活NK細(xì)胞，殺傷CD33+MDSCGleason M K, Ross J A, Warlick E D, et al. CD16xCD33 bisp

23、ecific killer cell engager (BiKE) activates NK cells against primary MDS and MDSC CD33+ targetsJ. Blood, 2014, 123(19): 3016-3026.BiTE與免疫檢查點(diǎn)抑制劑的聯(lián)合Khnke T, Krupka C, Tischer J, et al. Increase of PD-L1 expressing B-precursor ALL cells in a patient resistant to the CD19/CD3-bispecific T cell engager antibody blinatumomabJ. Journal of hematology & oncology, 2015, 8(1): 1-5.Osada T, Patel S P, Hammond S A, et al. CEA/CD3-bispecific T cell-engaging (BiTE) antibody-mediated T lymphocyte cytotoxicity maximized by inhibition of