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CN-122005781-A - Coagulation Factor XI (FXI) binding proteins

CN122005781ACN 122005781 ACN122005781 ACN 122005781ACN-122005781-A

Abstract

The invention relates to the field of medical biology, and discloses a single domain antibody aiming at a blood coagulation Factor XI (FXI) and a derivative protein thereof. In particular, the invention discloses a Factor XI (FXI) binding protein derived from a single domain antibody against Factor XI (FXI) and uses thereof.

Inventors

  • XU TING
  • WANG POPO
  • JIN YUHAO
  • GUO KANGPING
  • WANG LING
  • PANG MINJIE
  • WANG BILIN

Assignees

  • 苏州康宁杰瑞生物科技有限公司

Dates

Publication Date
20260512
Application Date
20210702
Priority Date
20200703

Claims (10)

  1. 1. Use of a composition comprising a Factor XI (FXI) binding protein and a pharmaceutically acceptable carrier for the manufacture of a medicament for the treatment and/or prevention of thromboembolism, inhibition of activation of FXI, or inhibition of blood coagulation and related thrombosis without compromising hemostasis in a subject in need thereof, wherein said FXI binding protein comprises at least one immunoglobulin single variable domain capable of specifically binding FXI comprising CDR1, CDR2, CDR3 as set forth in CDR1, CDR2 and CDR3 in a VHH as set forth in any one of SEQ ID NO 14, 4 and 10, Wherein the CDR is a Kabat CDR, an AbM CDR, a Chothia CDR, or an IMGT CDR.
  2. 2. The use according to claim 1, wherein CDR1, CDR2 and CDR3 of the VHH shown in SEQ ID NO. 14 are selected from any one of the group consisting of the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 180-182 respectively when the CDR is a Kabat CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 183-185 respectively when the CDR is an AbM CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 186-188 respectively when the CDR is a Chothia CDR, and the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 189-191 respectively when the CDR is an IMGT CDR.
  3. 3. The use according to claim 2, wherein the amino acid sequence of said at least one immunoglobulin single variable domain consists of the amino acid sequence shown in one of SEQ ID NOs 14, 318-323.
  4. 4. The use according to claim 1, wherein CDR1, CDR2 and CDR3 of the VHH shown in SEQ ID NO. 4 are selected from any one of the group consisting of the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 60-62 when the CDR is a Kabat CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 63-65 when the CDR is an AbM CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 66-68 when the CDR is a Chothia CDR, and the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO. 69-71 when the CDR is an IMGT CDR.
  5. 5. The use according to claim 4, wherein the amino acid sequence of said at least one immunoglobulin single variable domain consists of the amino acid sequence shown in one of SEQ ID NOs 4, 306-311.
  6. 6. The use according to claim 1, wherein CDR1, CDR2 and CDR3 of the VHH shown in SEQ ID NO 10 are selected from any one of the group consisting of the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO 132-134 respectively when the CDR is a Kabat CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO 135-137 respectively when the CDR is an AbM CDR, the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO 138-140 respectively when the CDR is a Chothia CDR, and the sequences of CDR1, CDR2 and CDR3 shown in SEQ ID NO 141-143 respectively when the CDR is an IMGT CDR.
  7. 7. The use according to claim 6, wherein the amino acid sequence of said at least one immunoglobulin single variable domain consists of the amino acid sequence shown in one of SEQ ID NOs 10, 312-317.
  8. 8. The use of any one of claims 1-7, wherein the FXI binding protein binds to the Apple2 domain of FXI.
  9. 9. The use of any one of claims 1-7, wherein the FXI binding protein further comprises an immunoglobulin Fc region.
  10. 10. The use of claim 9, wherein the immunoglobulin Fc region is a human immunoglobulin Fc region.

Description

Coagulation Factor XI (FXI) binding proteins The present application is a divisional application of patent application of the application named blood coagulation Factor XI (FXI) binding protein with the application number 202180047622.8 and the application number 2021, 7 and 2. Technical Field The invention relates to the field of medical biology, and discloses a single domain antibody aiming at a blood coagulation Factor XI (FXI) and a derivative protein thereof. In particular, the invention discloses a Factor XI (FXI) binding protein derived from a single domain antibody against Factor XI (FXI) and uses thereof. Background Coagulation factors are various protein components involved in the blood coagulation process. Its physiological role is to be activated when the blood vessel bleeds, adhere to the platelets and fill the leak in the blood vessel. This process is known as coagulation. They are produced in part by the liver. Can be inhibited by coumarin. For unified naming, the world health organization is numbered with roman numerals in the order in which it was found, blood clotting factors I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, etc. Factor XI (FXI) is a dimer composed of identical 80 KDa subunits, and each subunit consists of four Apple domains (A1, A2, A3 and A4) and a catalytic domain starting from the N-terminus. FXI is a zymogen that circulates in complex with high molecular weight kininogen (HK). HK binds to the A2 domain in FXI and is a physiological cofactor for FXIIa activation of FXI to FXIa. The remaining apple domains in FXI also mediate important physiological functions. For example, FIX binding ectosite is located in A3, whereas FXIIa binding site is in A4. Residues critical for FXI dimerization are also located in A4. Studies have shown that FXI plays a key role in the pathological process of thrombosis, contributes relatively little to hemostasis, and is therefore a promising target for thrombosis. In Ionis Pharmaceuticals inc. FXI antisense oligonucleotide (ASO) phase II trial (Buller et al, N Engl J Med 2015, 372:232-240), FXI ASO resulted in significant reduction of Venous Thromboembolism (VTE) with less tendency to bleed compared to enoxaparin in patients undergoing total knee arthroplasty. Human genetics and epidemics studies (Duga et al, semin Thromb Hemost 2013; chen et al, drug Discov Today 2014; key, hematology Am Soc Hematol Educ Program 2014, 2014:66-70) indicate that severe FXI deficiency (hemophilia C) reduces the risk of ischemic stroke and deep vein embolism, whereas increased FXI levels are associated with higher risk of VTE and ischemic stroke. Furthermore, many preclinical studies have shown that FXIa inhibition or loss of function mediates thromboprotection without compromising hemostasis (Chen et al Drug Discov Today 2014). Notably, monoclonal antibodies 14E11 and 1A6 produced significant thrombus reduction in the baboon AV branch thrombus formation model (U.S. Pat. No. 8,388,959; tucker et al, blood 2009, 113:936-944; cheng et al, blood 2010, 116:3981-3989). In addition, 14E11 (in that it cross-reacts with mouse FXI) provides protection in experimental models of mouse acute ischemic stroke (Leung et al Transl Stroke Res 2012, 3:381-389). Additional FXI-targeting mAb studies in preclinical models have also been reported, which confirm FXI as an antithrombotic target with minimal bleeding risk (van Montfoort et al, thromb Haemost 2013, 110; takahashi et al, , Thromb Res 2010, 125:464-470; van Montfoort, Ph.D. Thesis, University of Amsterdam, Amsterdam, Netherlands, 2014, 11, 14). Inhibition of FXI is thus a promising strategy for novel antithrombotic therapies with improved benefit-risk ratio compared to current standard anticoagulants. Brief Description of Drawings FIG. 1 shows FXI blocking activity of FXI single domain antibody Fc fusion protein (ATPP assays). FIG. 2 shows the blocking activity of the humanized FXI single domain antibody Fc fusion protein on FXI (ATPP assays). FIG. 3 shows the inhibitory effect of huFE bispecific antibody Fc fusion proteins on human FXI activity. FIG. 4 shows the inhibitory activity of huFE bispecific antibody Fc fusion proteins on APTT in human whole blood plasma. Fig. 5 shows the inhibitory activity of huFE bispecific antibody Fc fusion proteins on APTT of monkey whole blood plasma. FIG. 6 shows the inhibitory activity of huFE bispecific antibody Fc fusion proteins on APTT in rabbit whole blood plasma. FIG. 7 shows the effect of FXI single domain antibody Fc fusion protein and bispecific antibody on rabbit venous thrombosis. Detailed Description Definition of the definition Unless otherwise indicated or defined, all terms used have the usual meaning in the art, which will be understood by those skilled in the art. Reference is made, for example, to standard manuals such as Sambrook et al, "Molecular Cloning: A Laboratory Manual" (2 nd edition), volumes 1-3 , Cold Spring Harbor Laboratory Press (1989);Lewin,"Genes IV",Ox