TWI834254B - α-烯醇酶拮抗劑治療纖維化疾病之用途 - Google Patents
α-烯醇酶拮抗劑治療纖維化疾病之用途 Download PDFInfo
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Abstract
本揭示內容係有關一有效量之α-烯醇酶(烯醇酶-1,ENO-1)拮抗劑在製造用於治療纖維化疾病之藥劑的用途。ENO-1拮抗劑顯著減弱體重下降及肺重量增加以及肺纖維化病變與肺膠原沉積。同時,ENO-1拮抗劑顯著降低細胞遷移以及初代小鼠肺肌纖維母細胞中膠原與TGF-β的分泌。
Description
本揭示內容為纖維化疾病及結締組織疾病的領域。更具體而言,本揭示內容係有關α-烯醇酶(烯醇酶-1,ENO-1)拮抗劑用於治療及/或預防纖維化疾病,特別是特發性肺纖維化的用途。
纖維化疾病涉及在修復性或反應性過程中於器官或組織中形成過量的纖維結締組織。此類纖維化疾病包括特發性肺纖維化(idiopathic pulmonary fibrosis,IPF)、肺高血壓、肺纖維化、氣腫、非酒精性脂肪性肝炎、胰纖維化、腸纖維化、心臟纖維化、骨髓纖維化、關節纖維化、間質性肺病、非特異性間質性肺炎(non-specific interstitial pneumonia,NSIP)、尋常性間質性肺炎(usual interstitial pneumonia,UIP)、心肌內膜纖維化、縱隔纖維化、腹膜後纖維化、進行性大塊型纖維化(煤工塵肺症併發症)、腎因性全身性纖維化、克隆氏症(Crohn's disease)、陳舊性心肌梗塞、硬皮病/全身性硬化症、神經纖維瘤病、哈布二氏症候群(Hermansky-Pudlak syndrome)、糖尿病腎病變、腎纖維化、肥厚性心肌症(hypertrophic cardiomyopathy,HCM)、高血壓相關腎病、局部節段型腎絲球硬化症(focal segmental glomerulosclerosis,FSGS)、輻射誘導性纖維化、子宮肌瘤(纖維樣)、酒精性肝病、肝脂肪變性、肝纖維化、肝硬化、C型肝炎(HCV)感染、慢性器官移植排斥反應、皮膚纖維化狀況、瘢痕疙瘩(keloid scarring)、杜普宜特朗氏攣縮(Dupuytren contracture)、艾登二氏症候群(Ehlers-Danlos syndrome)、失養性表皮分解性水皰病(epidermolysis bullosa dystrophica)、口腔黏膜下纖維化,以及纖維增殖性疾病。
ENO-1為一種多功能蛋白,首先被發現作為醣解作用途徑的關鍵酵素。在正常情況下,ENO-1表達在胞質液中。然而,ENO-1亦被發現表達在許多癌細胞的細胞表面上(以作為纖維蛋白溶酶原受體)及活化型血液細胞(例如,嗜中性球、淋巴球及單核球)的細胞表面上。已知上調纖維蛋白溶酶原受體蛋白可誘導尿激酶纖維蛋白溶酶原活化系統的瀑流反應並造成細胞外基質降解。
本申請案係申請補充美國暫時專利申請號63/235,486於2021年8月20日提申之內容,出於所有目的,在此併入本案以作為參考資料。
本揭示內容係有關一種靶向ENO-1之α-烯醇酶(烯醇酶-1,ENO-1)拮抗劑及其用途,其中ENO-1拮抗劑具有與ENO-1的結合能力,例如人類ENO-1抗體,以作為抗原結合結構域,以便中和ENO-1的生物作用。ENO-1拮抗劑可結合至游離的ENO-1蛋白及細胞表面上的ENO-1蛋白,且在纖維化疾病的治療中具有重要應用前景。
根據本揭示內容之某些具體實施例,纖維化疾病或失調可為任何由ENO-1蛋白之異常活化或表達引起的情況。此類疾病之實例包括肝纖維化、腸纖維化、腎纖維化、皮膚纖維化、表皮纖維化、內皮纖維化、肌肉纖維化、肌腱纖維化、軟骨纖維化、心臟纖維化、胰纖維化、肺纖維化、子宮纖維化、神經系統纖維化、睾丸纖維化、陰莖纖維化、卵巢纖維化、腎上腺纖維化、動脈纖維化、靜脈纖維化、結腸纖維化、腸(例如,小腸)纖維化、膽道纖維化、軟組織(例如,縱隔或腹膜後)纖維化、骨髓纖維化、關節纖維化、眼纖維化、胃纖維化或其組合。
根據本揭示內容之某些具體實施例,纖維化疾病或失調可包括特發性肺纖維化(IPF)、肺高血壓、肺纖維化、氣腫、非酒精性脂肪性肝炎、胰纖維化、腸纖維化、心臟纖維化、骨髓纖維化、關節纖維化、間質性肺病、非特異性間質性肺炎(NSIP)、尋常性間質性肺炎(UIP)、心肌內膜纖維化、縱隔纖維化、腹膜後纖維化、進行性大塊型纖維化(煤工塵肺症併發症)、腎因性全身性纖維化、克隆氏症、陳舊性心肌梗塞、硬皮病/全身性硬化症、神經纖維瘤病、哈布二氏症候群(Hermansky-Pudlak syndrome)、糖尿病腎病變、腎纖維化、肥厚性心肌症(HCM)、高血壓相關腎病、局部節段型腎絲球硬化症(FSGS)、輻射誘導性纖維化、子宮肌瘤(纖維樣)、酒精性肝病、肝脂肪變性、肝纖維化、肝硬化、C型肝炎(HCV)感染、慢性器官移植排斥反應、皮膚纖維化狀況、瘢痕疙瘩(keloid scarring)、杜普宜特朗氏攣縮(Dupuytren contracture)、艾登二氏症候群(Ehlers-Danlos syndrome)、失養性表皮分解性水皰病(epidermolysis bullosa dystrophica)、口腔黏膜下纖維化,以及纖維增殖性疾病。在一較佳具體實施例中,纖維化疾病或失調可包括特發性肺纖維化、肺高血壓、氣腫、非酒精性脂肪性肝炎、胰纖維化、腎纖維化、腸纖維化、心臟纖維化、骨髓纖維化、關節纖維化或全身性硬化症。
根據本揭示內容之某些具體實施例,ENO-1拮抗劑可為一抗ENO-1抗體或其結合片段。根據本揭示內容之某些具體實施例,ENO-1拮抗劑可為一抗ENO-1嵌合抗原受體(CAR),其包含一抗原結合結構域、一鉸鏈區、一跨膜結構域及一傳訊結構域;其中抗原結合結構域至少為抗ENO-1抗體之一部分。
根據本揭示內容之某些具體實施例,抗原結合結構域可包含SEQ ID NO: 1(GYTFTSCVMN)、SEQ ID NO: 2(YINPYNDGTKYNEKFKG)、SEQ ID NO: 3(EGFYYGNFDN)、SEQ ID NO: 4(RASENIYSYLT)、SEQ ID NO: 5(NAKTLPE)及SEQ ID NO: 6(QHHYGTPYT)中所示之胺基酸序列。較佳地,抗體可包含一具有三個互補區的重鏈可變結構域,其包括HCDR1(GYTFTSCVMN;SEQ ID NO: 1)、HCDR2(YINPYNDGTKYNEKFKG;SEQ ID NO: 2)及HCDR3(EGFYYGNFDN;SEQ ID NO: 3);以及一具有三個互補區的輕鏈可變結構域,其包括LCDR1(RASENIYSYLT;SEQ ID NO: 4)、LCDR2(NAKTLPE;SEQ ID NO: 5)及LCDR3(QHHYGTPYT;SEQ ID NO: 6)。
根據本揭示內容之其他具體實施例,抗原結合結構域可包含SEQ ID NO: 7(GYTFTSXVMN,其中X為除了半胱胺酸以外的任何胺基酸)、SEQ ID NO: 2(YINPYNDGTKYNEKFKG)、SEQ ID NO: 3(EGFYYGNFDN)、SEQ ID NO: 4(RASENIYSYLT)、SEQ ID NO: 5(NAKTLPE)及SEQ ID NO: 6(QHHYGTPYT)中所示之胺基酸序列。較佳地,抗體可包含一具有三個互補區的重鏈可變結構域,其包括HCDR1(GYTFTSXVMN,其中X為除了半胱胺酸以外的任何胺基酸;SEQ ID NO: 7)、HCDR2(YINPYNDGTKYNEKFKG;SEQ ID NO: 2)及HCDR3(EGFYYGNFDN;SEQ ID NO: 3);以及一具有三個互補區的輕鏈可變結構域,其包括LCDR1(RASENIYSYLT;SEQ ID NO: 4)、LCDR2(NAKTLPE;SEQ ID NO: 5)及LCDR3(QHHYGTPYT;SEQ ID NO: 6)。
根據本揭示內容之某些具體實施例,鉸鏈區可包含一CD8 α 鉸鏈區(TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHT RGLDFACD;SEQ ID NO: 8);可選地,跨膜結構域可包含一CD8 α跨膜區(IYIWAPLAGTCGVLLLSLVIT;SEQ ID NO:9)及/或一CD28跨膜區(FWVLVVVGGVLACYSLLVTVAFIIFWV;SEQ ID NO: 10);可選地,傳訊結構域可包含CD3 ζ(RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR;SEQ ID NO:11);可選地,傳訊結構域可進一步包含4-1BB、CD28之細胞內區、DAP10、OX 40或其組合;可選地,抗ENO-1 CAR可進一步包含一訊息胜肽;可選地,訊息胜肽可具有一IgG κ輕鏈訊息胜肽、一CD8 α訊息胜肽、一GM-CSF訊息胜肽、一HSA訊息胜肽、一IgG重鏈訊息胜肽、一IgG輕鏈訊息胜肽、一CD33訊息胜肽、一IL-2訊息胜肽或一胰島素訊息胜肽。
根據本揭示內容之某些具體實施例,抗ENO-1 CAR可具有一訊息胜肽、一抗ENO-1抗體、一CD8 α鉸鏈區、一CD8 α跨膜區、4-1BB及CD3 ζ。
根據本揭示內容之某些具體實施例,ENO-1拮抗劑可為一特異性地結合至ENO-1的免疫結合體,其包含:通式Ab-(L-D)m (I),其中Ab為一抗ENO-1抗體或其結合片段,L為一連接子或一直接的鍵,D為一治療劑或一標記,且m為一從1至12的整數。較佳地,抗體可為一單株抗體。較佳地,抗體可為一小鼠抗體、一人類抗體、一嵌合抗體、一人源化抗體或其抗體片段。
根據本揭示內容之某些具體實施例,治療劑可具有一抗纖維化劑、免疫調節劑、放射性同位素及毒素。較佳地,ENO-1拮抗劑可與至少一具有已知抗纖維化活性之治療上活性劑組合使用,該活性劑選自於吡非尼酮(pirfenidone)或受體酪胺酸激酶抑制劑(RTKI),例如尼達尼布(Nintedanib)、索拉非尼(Sorafenib)及其他RTKI,或血管收縮素II型(AT1)受體阻斷劑,或CTGF抑制劑,或任何易於干擾TGF-β與BMP活化途徑的抗纖維化化合物,其包括潛伏TGF-β複合物的活化劑,例如MMP2、MMP9、THBS1或細胞表面整合素,TGF3受體I型(TGFBRI)或II型(TGFBRII)及其等之配體,例如TGF3、活化素、抑制素、Nodal、抗穆勒氏荷爾蒙(anti-Mullerian hormone)、GDF或BMP,輔助共受體(亦稱為III型受體),或SMAD依賴性經典途徑的成分,其包括調節性或抑制性SMAD蛋白,或SMAD非依賴性或非經典途徑的成員,其包括MAPK傳訊的各種分支,TAK1傳訊途徑、Rho樣GTPase傳訊途徑、磷脂酸肌醇-3激酶/AKT途徑、TGF-β誘導的EMT過程或經典與非經典刺蝟蛋白(Hedgehog)傳訊途徑,其包括Hh配體或標的基因,或任何WNT的成員,或Notch途徑,其易於影響TGF-β傳訊。
根據本揭示內容之某些具體實施例,ENO-1拮抗劑可透過口服、腸胃外、口頰、陰道、直腸、吸入、吹入、舌下、肌肉內、皮下、局部、鼻內、腹腔內、胸腔內、靜脈內、硬膜外、鞘內或腦室內途徑投予,或透過注射至關節中。
根據本揭示內容之某些具體實施例,受試者為一哺乳動物。在一較佳之具體實施例中,受試者為人類。
根據本揭示內容之某些具體實施例,ENO-1拮抗劑可為一核酸,例如DNA或RNA,其被設計成遞輸至細胞中並表達為細胞內之胜肽蛋白。舉例而言,ENO-1拮抗劑可為能轉錄及轉譯為例如抗ENO-1抗體或其結合片段的核酸。此外,核酸可具有或不具有一分泌訊息胜肽,使得由核酸轉錄及轉譯的蛋白或胜肽可為分泌型、非分泌型或其組合。根據本揭示內容之某些具體實施例,核酸可經由任何已知方法或媒介遞輸至受試者,例如病毒載體、聚合物或微脂體。根據本揭示內容之某些具體實施例,核酸可以已知經修飾之核苷酸取代,例如偽UTP、1-Me偽UTP、5-甲氧基UTP、N1-乙基偽UTP、5-甲基CTP或N4-乙醯基-CTP,以提高表達效率。
相較於先前技術,本發明具有下列有利的效果。根據本揭示內容,體內與體外實驗證實ENO-1拮抗劑(例如,ENO-1 mAb HuL217)的抗纖維化功效及抗發炎功效。亦即,ENO-1拮抗劑為纖維化疾病(例如,IPF)的潛在療法。
所屬領域中具有通常知識者將理解,藥學上有效量取決於許多因素,例如患者狀況、年齡、疾病狀態、投予途徑等,且此類有效量可在常規實踐中基於彼等因素確定,而無需進行過多的實驗。
本發明之其他態樣將由於下列描述而變得顯而易見。
一般定義
除非另有指明,否則本發明之實踐將採用分子生物學、微生物學、重組DNA及免疫學的常規技術,彼等技術在本領域之技術範圍內。此類技術在文獻中充分解釋。參見,例如,Molecular Cloning A Laboratory Manual,第 2 版,由Sambrook、Fritsch及Maniatis編輯(Cold Spring Harbor Laboratory Press,1989);DNA Cloning,第I卷與第II卷(D. N. Glover編輯,1985);Culture Of Animal Cells(R. I. Freshney,Alan R. Liss, Inc.,1987);Immobilized Cells And Enzymes(IRL Press,1986);B. Perbal,A Practical Guide To Molecular Cloning(1984);論文,Methods In Enzymology(Academic Press, Inc.,N.Y.);Gene Transfer Vectors For Mammalian Cells(J. H. Miller與M. P. Calos編輯,1987,Cold Spring Harbor Laboratory);Methods In Enzymology,第154卷與第155卷(Wu等人編輯),Immunochemical Methods In Cell And Molecular Biology(Mayer與Walker編輯,Academic Press,London,1987);Antibodies: A Laboratory Manual,由Harlow and Lane所著(Cold Spring Harbor Laboratory Press,1988);以及Handbook Of Experimental Immunology,第I-IV卷(D. M. Weir與C. C. Blackwell編輯,1986)。
術語「抗體」與「免疫球蛋白」在最廣義上可互換使用,包括單株抗體(例如,全長或完整單株抗體)、多株抗體、單價、多價抗體、多特異性抗體(例如,雙特異性抗體,只要其等表現出所需的生物活性),且亦可包括某些抗體片段(如本文更詳細之描述)。抗體可為嵌合的、人類的、人源化的及/或親和力成熟的。
術語「可變」意指可變結構域的某些部分在抗體之序列中有很大差異,並用於每一特定抗體對其特定抗原的結合及特異性。然而,可變性並非均勻地分佈在抗體的可變結構域中。其集中在三個部分,稱為輕鏈與重鏈可變結構域之互補決定區(CDR)或高度可變區。可變結構域的更高度保留部分稱為框架(FR)。原始重鏈與輕鏈的可變結構域各包含四個FR區,主要採用β摺疊構形,由三個CDR連接,其形成連接β摺疊結構的圈環,且在一些情況下形成β摺疊結構的一部分。每一鏈中之CDR由FR區緊密相連,並與來自另一鏈的CDR一起協助形成抗體的抗原結合位點(參見Kabat等人,Sequences of Proteins of Immunological Interest,第五版,National Institute of Health,Bethesda,Md.(1991))。恆定結構域不直接參與抗體與抗原的結合,但表現出各種效應子功能,例如以抗體依賴性細胞毒性的抗體參與。
抗體可為全長抗體或可包含具有抗原結合部分之抗體的一個片段(或多個片段),包括但不侷限於,Fab、F(ab')
2、Fab'、F(ab)'、Fv、單鏈Fv (scFv)、二價scFv (bi-scFv)、三價scFv (tri-scFv)、Fd、dAb片段(例如,Ward等人,Nature,341:544-546(1989))、CDR、雙抗體、三抗體、四抗體、線性抗體、單鏈抗體分子,以及由抗體片段形成的多特異性抗體。本發明亦包括透過使用重組方法或合成連接子連接抗體片段而產生的單鏈抗體。Bird等人,Science,1988,242:423-426。Huston等人,Proc. Natl. Acad. Sci. USA,1988,85:5879-5883。
如本文所用,「治療」意指試圖改變欲治療之個體或細胞之自然過程的臨床介入,並可用於預防或在臨床病理學過程中進行。治療的所需效果包括預防疾病的發生或復發、減輕症狀、減少疾病的任何直接或間接病理後果、預防或減少發炎及/或組織/器官損傷、降低疾病進展速度、改善或緩解疾病狀態,以及緩解或改善預後。在一些具體實施例中,本揭示內容之抗體用於延遲疾病或失調的發展。
「個體」或「受試者」為一脊椎動物。在某些具體實施例中,脊椎動物為一哺乳動物。哺乳動物包括但不侷限於,農場動物(例如,牛)、運動動物、寵物(例如,貓、狗及馬)、靈長類動物、小鼠及大鼠。在某些具體實施例中,脊椎動物為一人類。
「有效量」意指在必要的劑量與時間段內有效達到所需治療或預防結果的量。然而,本揭示內容之物質/分子的「有效量」可根據例如個體的疾病狀態、年齡、性別及體重以及物質/分子的能力等因素而變,以引發個體所需的反應。有效量亦為物質/分子之任何毒性或有害作用透過治療上有益效果所抵消的量。在一具體實施例中,抗ENO-1抗體之有效量範圍為1-1000 mg/kg,較佳為5-100 mg/kg,更佳為10-50 mg/kg,例如5、10、15、20、30、40、50、60、70、80、90或100 mg/kg。
本文中使用之術語「治療劑」意指抑制或防止細胞功能及/或導致細胞破壞的物質。該術語旨在包括抗纖維化劑、放射性同位素(例如,
211At、
131I、
125I、
90Y、
186Re、
188Re、
153Sm、
212Bi、
32P、
60C,以及鎦-177、鍶-89及釤(
153Sm)的放射性同位素)、免疫調節劑,以及毒素,例如細菌、真菌、植物或動物源的小分子毒素或酵素上活性毒素,包括其合成類似物及衍生物。
「纖維化病症」、「纖維增殖性病症」、「纖維化疾病」、「纖維增殖性疾病」、「纖維化失調」以及「纖維增殖性失調」可互換使用以意指病症、疾病或失調,其特徵為纖維母細胞之增殖失調或活性失調,及/或膠原組織之病理性累積或過度累積。典型上,任何此類疾病、失調或病症皆可透過投予具有抗纖維化作用之化合物進行治療。纖維化疾病包括但不侷限於,特發性肺纖維化(IPF)、肺高血壓、肺纖維化、氣腫、非酒精性脂肪性肝炎、胰纖維化、腸纖維化、心臟纖維化、骨髓纖維化、關節纖維化、間質性肺病、非特異性間質性肺炎(NSIP)、尋常性間質性肺炎(UIP)、心肌內膜纖維化、縱隔纖維化、腹膜後纖維化、進行性大塊型纖維化(煤工塵肺症併發症)、腎因性全身性纖維化、克隆氏症、陳舊性心肌梗塞、硬皮病/全身性硬化症、神經纖維瘤病、哈布二氏症候群(Hermansky-Pudlak syndrome)、糖尿病腎病變、腎纖維化、肥厚性心肌症(HCM)、高血壓相關腎病、局部節段型腎絲球硬化症(FSGS)、輻射誘導性纖維化、子宮肌瘤(纖維樣)、酒精性肝病、肝脂肪變性、肝纖維化、肝硬化、C型肝炎(HCV)感染、慢性器官移植排斥反應、皮膚纖維化狀況、瘢痕疙瘩(keloid scarring)、杜普宜特朗氏攣縮(Dupuytren contracture)、艾登二氏症候群(Ehlers-Danlos syndrome)、失養性表皮分解性水皰病(epidermolysis bullosa dystrophica)、口腔黏膜下纖維化,或纖維增殖性疾病。
本文中使用之術語「特發性肺纖維化」意指一種慢性、進行性且通常致命的肺部疾病,其被認為是慢性發炎過程的結果。
本文中使用之術語「抗纖維化劑」意指一已知具有抗纖維化作用的物質。本術語旨在包括吡非尼酮(pirfenidone)或受體酪胺酸激酶抑制劑(RTKI),例如尼達尼布(Nintedanib)、索拉非尼(Sorafenib)及其他RTKI,或血管收縮素II型(AT1)受體阻斷劑,或CTGF抑制劑,或任何易於干擾TGF-β與BMP活化途徑的抗纖維化化合物,其包括潛伏TGF-β複合物的活化劑,例如MMP2、MMP9、THBS1或細胞表面整合素,TGF3受體I型(TGFBRI)或II型(TGFBRII)及其等之配體,例如TGF3、活化素、抑制素、Nodal、抗穆勒氏荷爾蒙(anti-Mullerian hormone)、GDF或BMP,輔助共受體(亦稱為III型受體),或SMAD依賴性經典途徑的成分,其包括調節性或抑制性SMAD蛋白,或SMAD非依賴性或非經典途徑的成員,其包括MAPK傳訊的各種分支,TAK1傳訊途徑、Rho樣GTPase傳訊途徑、磷脂酸肌醇-3激酶/AKT途徑、TGF-β誘導的EMT過程或經典與非經典刺蝟蛋白(Hedgehog)傳訊途徑,其包括Hh配體或標的基因,或任何WNT的成員,或Notch途徑,其易於影響TGF-β傳訊。
本揭示內容之藥劑可局部或全身施加。本揭示內容之藥劑亦可以組合或與輔因子一起供應。若本揭示內容之藥劑正常存在於目標位置中,則可以足以恢復正常水平的量投予,或其等可以使目標位置之水平升至高於正常水平的量投予。
本揭示內容之藥劑可從外部來源供應至目標位置,或其等可透過目標位置中之細胞或與目標位置相同的生物體中之細胞在體內製造。
本揭示內容之藥劑可為任何生理上合適的製劑。其等可透過注射、局部、吸入、口服或任何其他有效方式投予生物體。
上述之阻斷或抑制過度纖維化形成與維持的相同藥劑及方法亦可用於阻斷或抑制不適當的纖維化形成。舉例而言,其等可治療或預防發生在肝、腎、肺、心臟與心包膜、眼睛、皮膚、口、胰腺、胃腸道、大腦、乳房、骨髓、骨骼、生殖泌尿道、腫瘤或傷口的病症。
一般而言,其等可治療或預防由包括但不侷限於類風濕性關節炎、狼瘡、致病性纖維化、纖維化疾病、纖維化病變(例如該等日本血吸蟲(Schistosoma japonicum)感染後形成的)、放射損傷、自體免疫性疾病、萊姆病、化療引起的纖維化、HIV或感染引起的局灶性硬化症、由於脊柱手術疤痕、腹部黏連術後疤痕及纖維囊性形成造成的背部手術失敗症候群(failed back syndrome)。
本揭示內容之具體實施例係有關含有ENO-1抗體之抗體-藥物結合體(antibody-drug conjugate)及其等在治療纖維化疾病之用途。ENO-1為多功能蛋白,其被發現表達在許多癌細胞的細胞表面上以作為纖維蛋白溶酶原受體以及表達在活化的造血細胞上,例如嗜中性球、淋巴球及單核球。因此,基於針對ENO-1之抗體的ADC可為適用的診斷劑及/或治療劑。
然而,治療性抗體之快速內化或缺乏ADCC活性可能造成抗體無效以及抗性。因此,需要增強以抗ENO-1為主之治療劑的療效。一方法為將有效負載與抗ENO-1抗體結合(亦即,抗體-藥物結合體)。
根據本揭示內容之具體實施例,抗ENO-1抗體或其結合片段可耦接至一藥物、診斷劑或一治療劑。因此,本文中所用之術語「抗體-藥物結合體(ADC)」可指耦接至一有效負載(其可為一藥物、一診斷劑或一治療劑)的抗體部分(其可為一全抗體或其結合片段)。示例性ADC係如WO 2021/228044 A1中所述,其內容整體併入本案以作為參考資料。
治療纖維化疾病之方法
本揭示內容提供治療纖維化疾病(例如,特發性肺纖維化(IPF))之方法。本方法一般而言涉及投予有需求之受試者一有效量之α-烯醇酶(烯醇酶-1,ENO-1)拮抗劑。
在一些具體實施例中,在本方法中使用的ENO-1拮抗劑可包括,但不侷限於:
(1) 一抗ENO-1抗體或其結合片段;
(2) 一抗ENO-1嵌合抗原受體(CAR),其包含一抗原結合結構域、一鉸鏈區、一跨膜結構域及一傳訊結構域;其中抗原結合結構域至少為抗ENO-1抗體之一部分;
(3) 一特異性地結合至ENO-1的免疫結合體,其包含:通式Ab-(L-D)m (I),其中Ab為一抗ENO-1抗體或其結合片段,L為一連接子或一直接的鍵,D為一治療劑或一標記,且m為一從1至12的整數;或
(4) 抗ENO-1抗體或其結合片段或抗ENO-1 CAR之核酸,其被遞輸至細胞中並表達為細胞內抗ENO-1抗體或其結合片段或抗ENO-1 CAR。
本揭示內容之具體實施例將以下列特定實施例說明。所屬領域中具有通常知識者將理解,彼等實施例僅用於說明,且可能進行其他修正及改變而不背離本揭示內容之範疇。
實施例
實施例
1.
抗
ENO-1
抗體之製備
根據本揭示內容之具體實施例,產生抗ENO-1抗體的一般方法包括獲得產生針對ENO-1之單株抗體的融合瘤。產生單株抗體的方法為本領域已知,在此將不詳述。簡言之,以抗原(ENO-1)配合適當的佐劑挑戰小鼠。隨後,收穫免疫小鼠的脾細胞並與融合瘤融合。使用任何已知之方法,例如ELISA,可鑑定出具有ENO-1抗原結合能力的陽性殖株。在一具體實施例中,抗ENO-1抗體為HuL217。
抗體-藥物結合體(ADC)可特異性地靶向ENO-1。彼等ADC可使用任何特異性地結合至ENO-1的抗體。舉例而言,本發明之ADC可使用一小鼠或人源化抗ENO-1抗體,或其scFv或Fab片段。示例性抗ENO-1抗體,例如HuL217,可包含一具有三個互補區的重鏈可變結構域,其包括HCDR1(GYTFTSCVMN;SEQ ID NO: 1)、HCDR2(YINPYNDGTKYNEKFKG;SEQ ID NO: 2)及HCDR3(EGFYYGNFDN;SEQ ID NO: 3),以及一具有三個互補區的輕鏈可變結構域,其包括LCDR1(RASENIYSYLT;SEQ ID NO: 4)、LCDR2(NAKTLPE;SEQ ID NO: 5)及LCDR3(QHHYGTPYT;SEQ ID NO: 6)。另一示例性抗ENO-1抗體可包含一具有三個互補區的重鏈可變結構域,其包括HCDR1(GYTFTSXVMN,其中X為除了半胱胺酸以外的任何胺基酸;SEQ ID NO: 7)、HCDR2(YINPYNDGTKYNEKFKG;SEQ ID NO: 2)及HCDR3(EGFYYGNFDN;SEQ ID NO: 3),以及一具有三個互補區的輕鏈可變結構域,其包括LCDR1(RASENIYSYLT;SEQ ID NO: 4)、LCDR2(NAKTLPE;SEQ ID NO: 5)及LCDR3(QHHYGTPYT;SEQ ID NO: 6)。
根據本揭示內容之具體實施例,抗體可為小鼠抗體。或者,抗體可為嵌合抗體(例如,耦接至小鼠可變區的人類恆定區)或人源化抗體(例如,接枝在人類免疫球蛋白之框架區上的小鼠CDR)或完全地人類抗體。
單株抗體可被人源化,其係透過從融合瘤獲得CDR序列並將CDR序列選殖至人類框架序列中以產生人源化抗體。可使用本領域已知之任何用於鑑定CDR序列的常用方法。本發明之CDR區係以Kabat編號格式進行識別。首先,產生抗ENO-1之融合瘤(例如,小鼠融合瘤)。此融合瘤可以標準方案產生,以產生單株抗體。隨後,使用例如TRIzol
®試劑,分離融合瘤之總RNA。隨後,使用例如第一股cDNA合成套組(Superscript III)及寡(dT
20)引子或Ig-3’恆定區引子,從總RNA合成cDNA。
隨後,從cDNA選殖免疫球蛋白基因之重鏈與輕鏈可變區。舉例而言,透過PCR,使用小鼠Ig-5’引子組(Novagen),從小鼠融合瘤cDNA擴增抗ENO-1 mAb之VH與VL可變區。使用CloneJet
TMPCR選殖套組(Ferments),PCR產物可直接選殖至適用之載體(例如,pJET1.2載體)中。pJET1.2載體含有致死插入,且僅在所需基因被選殖至該致死區中時才能在選擇條件下存活。此促進重組菌落的篩選。最終,從重組菌落中篩選出所需的殖株,將該等殖株之DNA分離並定序。可在國際ImMunoGeneTics資訊系統(IGMT)網站上分析免疫球蛋白(IG)核苷酸序列。
抗體表達及純化
針對抗體產生,經分離之殖株可在任何適用之細胞中表達。作為一實例,F293細胞(Life technologies)以抗ENO-1 mAb表達質體轉染並培養7天。使用蛋白A親和力管柱(GE),從培養基中純化抗ENO-1抗體。使用本領域已知之程序或根據製造商之說明,以Bio-Rad蛋白試驗套組測定蛋白濃度,並以12% SDS-PAGE分析。
實施例
2.
人類與小鼠纖維化肺中
ENO-1
之過度表達
在此實施例中,以ENO-1免疫組織化學(IHC)染色確定ENO-1是否在纖維化肺中異常表達。在圖1A中,福爾馬林固定、石蠟包埋(FFPE)的肺組織樣品係購自商業來源。三個正常人類肺FFPE組織切片係獲自BioChain Institute, Inc.(Newark,CA,USA)及US Biomax(Derwood,MD,USA)。三種人類纖維化肺FFPE切片係獲自OriGene Technologies(Rockville,MD,USA)。發現在人類纖維化肺中而非正常肺中的ENO-1表達升高。定量結果顯示於圖1B中。
博來黴素誘導的肺纖維化C57BL/6小鼠為人類纖維化疾病特發性肺纖維化最常用的實驗模型。7至9週大的雄性C57BL/6小鼠以氣管內方式給予單劑的博來黴素(3 mg/kg)。在21天後,收穫肺以進行ENO-1 IHC染色。圖1C說明相較於假實驗組,ENO-1在博來黴素組(BLM)中之小鼠肺部有過度表達。定量結果顯示於圖圖1D。
實施例
3. ENO-1
抗體用於
IPF
疾病模型
於博來黴素誘導的肺纖維化C57BL/6小鼠評估HuL217之作用。7至9週大的雄性C57BL/6小鼠以氣管內方式給予單劑的博來黴素(3 mg/kg)。將小鼠隨機分成三組,分別為假實驗組4隻小鼠、博來黴素 + 載體7隻小鼠,或博來黴素 + HuL217組7隻小鼠。將博來黴素的挑戰日定為第0天。HuL217組的小鼠在第1、7、13及19天進行靜脈注射。結果說明,相較於博來黴素組,以HuL217治療能減弱:體重下降、肺重量增加(圖2)、阿希克夫分數、發炎分數(圖3),以及肺膠原含量與支氣管肺泡灌洗液(bronchial alveolar lavage fluid,BALF)中之TGF-β的量(圖4)。此外,透過HuL217治療,減少了博來黴素誘導小鼠肺中肌纖維母細胞的累積(圖5)。為了研究HuL217的抗發炎作用,使用流式細胞術,分析了在BALF中單核球與嗜中性球募集進入肺的情形。圖6證實HuL217(在博來黴素注射前2小時進行靜脈注射)在第4天減少單核球與嗜中性球募集進入博來黴素誘導小鼠的肺中。
實施例
4. ENO-1
抗體對肺纖維母細胞之體外作用
初代小鼠肺纖維母細胞分離自雄性C57BL/6小鼠,且初代人類肺纖維母細胞購自Lonza(London,UK)。兩種細胞皆處理TGF-β以誘導遷移。圖7證實HuL217劑量依賴性地降低TGF-β處理之初代肺纖維母細胞的遷移能力。透過使用分離自正常(NHLF)或IPF患者(DHLF-IPF)的初代人類肺纖維母細胞,進一步證實HuL217的體外抗纖維化作用。HuL217可劑量依賴性地降低TGF-β刺激之NHLF(圖8)或DHLF-IPF(圖9)中膠原、TGF-β1及VEGF的分泌。
總之,在博來黴素小鼠中,HuL217顯著減弱體重下降、減弱肺重量之增加、以及減少肺纖維化病變與肺膠原沉積。升高的TGF-β與單核球的量在支氣管肺泡灌洗液中亦下降。HuL217可顯著降低初代小鼠肺肌纖維母細胞的細胞遷移及膠原與TGF-β分泌。
除非另有定義,否則本文使用的所有技術與科學術語以及任何縮寫字具有與所屬領域中具有通常知識者通常理解的相同含義。儘管與本文所述之該等相似或等效的用於溝通資訊的任何組合物、方法、套組及工具可用於實施本發明,但本文描述了用於溝通資訊的較佳組合物、方法、套組及工具。
本文引用的所有參考資料在法律允許的全部範圍內皆併入本案以作為參考資料。彼等參考資料的討論僅旨在總結該等作者的結論。未取用與先前技術相關的任何參考資料(或參考資料之一部分)。申請人保留質疑任何引用之參考資料的準確性與相關性的權利。
無
圖 1顯示人類纖維化肺及博來黴素(bleomycin)誘導的肺纖維化鼠科模型肺中ENO-1的過度表達。詳細內容描述於實施例2中。
圖 2顯示HuL217在博來黴素誘導的肺纖維化鼠科模型中減弱體重下降及肺重量增加的體內抗纖維化功效。詳細內容描述於實施例3中。
圖 3顯示ENO-1 mAb HuL217在博來黴素誘導肺纖維化鼠科模型的肺切片中減少阿希克夫分數(Ashcroft score)及發炎分數的體內抗纖維化功效。詳細內容描述於實施例3中。
圖 4顯示ENO-1 mAb HuL217在博來黴素誘導的肺纖維化鼠科模型中降低肺膠原及支氣管肺泡灌洗液中之TGF-β的體內抗纖維化功效。詳細內容描述於實施例3中。
圖 5顯示ENO-1 mAb HuL217在博來黴素誘導的肺纖維化鼠科模型中減少肺肌纖維母細胞(α-SMA陽性)的體內抗纖維化功效。詳細內容描述於實施例3中。
圖 6顯示ENO-1 mAb HuL217在博來黴素誘導的肺纖維化鼠科模型中減少肺單核球與肺嗜中性球募集的體內抗發炎功效。詳細內容描述於實施例3中。
圖 7顯示ENO-1 mAb HuL217在降低TGF-β刺激之初代小鼠與人類肺纖維母細胞遷移的體外抗纖維化功效。詳細內容描述於實施例4中。
圖 8顯示ENO-1 mAb HuL217在降低TGF-β1刺激之初代人類正常纖維母細胞中之膠原、TGF-β1及VEGF分泌的體外抗纖維化功效。詳細內容描述於實施例4中。
圖 9顯示ENO-1 mAb HuL217在降低初代人類IPF纖維母細胞中之膠原、TGF-β1及VEGF分泌的體外抗纖維化功效。詳細內容描述於實施例4中。
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TW202319066A_111131225_SEQL.xml
Claims (7)
- 一種一有效量之α-烯醇酶(烯醇酶-1,ENO-1)拮抗劑在製造用於治療肺纖維化疾病之藥劑的用途;其中該ENO-1拮抗劑為一抗ENO-1抗體,該抗ENO-1抗體包含:一具有三個互補區的重鏈可變結構域,其包括:如SEQ ID NO:1所示胺基酸序列的HCDR1,或如SEQ ID NO:7所示胺基酸序列的HCDR1,其中X為除了半胱胺酸以外的任何胺基酸,如SEQ ID NO:2所示胺基酸序列的HCDR2,以及如SEQ ID NO:3所示胺基酸序列的HCDR3;以及一具有三個互補區的輕鏈可變結構域,其包括:如SEQ ID NO:4所示胺基酸序列的LCDR1,如SEQ ID NO:5所示胺基酸序列的LCDR2,以及如SEQ ID NO:6所示胺基酸序列的LCDR3。
- 如請求項1之用途,其中該ENO-1拮抗劑為一特異性地結合至ENO-1的免疫結合體,其包含:通式Ab-(L-D)m (I),其中Ab為一抗ENO-1抗體或其結合片段,L為一連接子或一直接的鍵,D為一治療劑或一標記,且m為一從1至12的整數。
- 如請求項2之用途,其中該治療劑包含一抗纖維化劑、免疫調節劑、放射性同位素及毒素。
- 如請求項2之用途,其中該抗纖維化劑包含吡非尼酮(pirfenidone)或受體酪胺酸激酶抑制劑(RTKI),或血管收縮素II型(AT1)受體阻斷劑,或CTGF抑制劑,或干擾TGF-β與BMP活化途徑的抗纖維化化合物,或SMAD依賴性經典途徑的成分,或SMAD非依賴性或非經典途徑的成分。
- 如請求項2之用途,其中該標記包含一診斷劑或成像劑。
- 如請求項1之用途,其中該ENO-1拮抗劑係經由一病毒載體、一聚合物及/或微脂體遞輸。
- 如請求項1之用途,其中該肺纖維化疾病包含特發性肺纖維化(idiopathic pulmonary fibrosis,IPF)、間質性肺病、漸進性纖維化間質性肺病(progressive fibrosing interstitial lung disease,PF-ILD)、全身硬化症有關之間質性肺病(systemic sclerosis associated interstitial lung disease,SSc-ILD)、自體免疫疾病引起之間質性肺病或肺纖維化、肺高血壓、肺纖維化、肺氣腫、非特異性間質性肺炎(non-specific interstitial pneumonia,NSIP)、尋常性間質性肺炎(usual interstitial pneumonia,UIP)輻射誘導性肺纖維化、癌症引起之肺纖維化、藥物引起之肺纖維化、放射治療或化學治療引起之肺纖維化。
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CN106459181A (zh) * | 2013-12-20 | 2017-02-22 | 财团法人生物技术开发中心 | α‑烯醇化酶特异性抗体及其在免疫疾病治疗中的使用方法 |
CN106488932A (zh) * | 2013-12-20 | 2017-03-08 | 财团法人生物技术开发中心 | α‑烯醇化酶特异性抗体及其在癌症治疗中的使用方法 |
WO2018232372A1 (en) * | 2017-06-16 | 2018-12-20 | Protelica, Inc. | Fibronectin binding domain chimeric antigen receptors and methods of use thereof |
CN111093701A (zh) * | 2017-06-15 | 2020-05-01 | 财团法人生物技术开发中心 | 含有抗globo h抗体的抗体药物偶联物及其用途 |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN106459181A (zh) * | 2013-12-20 | 2017-02-22 | 财团法人生物技术开发中心 | α‑烯醇化酶特异性抗体及其在免疫疾病治疗中的使用方法 |
CN106488932A (zh) * | 2013-12-20 | 2017-03-08 | 财团法人生物技术开发中心 | α‑烯醇化酶特异性抗体及其在癌症治疗中的使用方法 |
CN111093701A (zh) * | 2017-06-15 | 2020-05-01 | 财团法人生物技术开发中心 | 含有抗globo h抗体的抗体药物偶联物及其用途 |
WO2018232372A1 (en) * | 2017-06-16 | 2018-12-20 | Protelica, Inc. | Fibronectin binding domain chimeric antigen receptors and methods of use thereof |
Non-Patent Citations (2)
Title |
---|
期刊 Jie Zhang et al.; Silencing of ENO1 inhibits the proliferation, migration and invasion of human breast cancer cells; JBUON; 25(2); 2020; 696-701 * |
期刊 Jie Zhang et al.; Silencing of ENO1 inhibits the proliferation, migration and invasion of human breast cancer cells; JBUON; 25(2); 2020; 696-701。 |
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