Anticoagulant antidotes

Abstract

The present invention relates to antibody molecules against anticoagulants, in particular dabigatran, and their use as antidotes of such anticoagulants.

What we claim: 
     
         1 . An antibody molecule capable of neutralizing the activity of an anticoagulant. 
     
     
         2 . The antibody molecule of  claim 1  wherein said antibody molecule has binding specificity for the anticoagulant. 
     
     
         3 . The antibody molecule of  claim 1  wherein the anticoagulant is a direct thrombin inhibitor, a Factor Xa inhibitor, or a vitamin K antagonist. 
     
     
         4 . The antibody molecule of  claim 3 , wherein the anticoagulant is dabigatran, argatroban, melagatran, ximelagatran, hirudin, bivalirudin, lepirudin, desirudin, apixaban, otamixaban, rivaroxaban, defibrotide, ramatroban, antithrombin III, or drotrecogin alpha. 
     
     
         5 . The antibody molecule of  claim 2 , wherein the anticoagulant is a disubstituted bicyclic heterocycle of general formula
   R a -A-Het-B-Ar-E,  (I)
   wherein   A denotes a carbonyl or sulphonyl group linked to the benzo, pyrido or thieno moiety of the group Het,   B denotes an ethylene group in which the methylene group linked to the group Ar may be replaced by an oxygen or sulphur atom or by an —NR 1 — group, wherein
 R 1  denotes a hydrogen atom or a C 1-4 -alkyl group, 
   E denotes an R b NH—C(═NH)— group wherein
 R b  denotes a hydrogen atom, a hydroxy, C 1-9 -alkoxycarbonyl, cyclohexyloxycarbonyl, phenyl-C 1-3 -alkoxycarbonyl, benzoyl, p-C 1-3 -alkyl-benzoyl or pyridinoyl group, whilst the ethoxy moiety in the 2-position of the abovementioned C 1-3 -alkoxycarbonyl group may additionally be substituted by a C 1-3 -alkylsulphonyl or 2-(C 1-3 -alkoxy)-ethyl group, 
   Ar denotes a 1,4-phenylene group optionally substituted by a chlorine atom or by a methyl, ethyl or methoxy group or it denotes a 2,5-thienylene group,   Het denotes a 1-(C 1-3 -alkyl)-2,5-benzimidazolylene, 1-cyclopropyl-2,5-benzimidazolylene, 2,5-benzothiazolylene, 1-(C 1-3 -alkyl)-2,5-indolylene, 1-(C 1-3 -alkyl)-2,5-imidazo[4,5-b]pyridinylene, 3-(C 1-3 -alkyl)-2,7-imidazo[1,2-a]pyridinylene or 1-(C 1-3 -alkyl)-2,5-thieno[2,3-d]imidazolylene group and   R a  denotes an R 2 NR 3 — group wherein
 R 2  is a C 1-4 -alkyl group which may be substituted by a carboxy, C 1-6 -alkyloxycarbonyl, benzyloxycarbonyl, C 1-3 -alkylsulphonylaminocarbonyl or 1H-tetrazol-5-yl group, 
 a C 2-4 -alkyl group substituted by a hydroxy, benzyloxy, carboxy-C 1-3 -alkylamino, C 1-3 -alkoxycarbonyl-C 1-3 -alkylamino, N—(C 1-3 -alkyl)-carboxy-C 1-3 -alkylamino or N—(C 1-3 -alkyl)-C 1-3 -alkoxycarbonyl-C 1-3 -alkylamino group, whilst in the abovementioned groups the carbon atom in the α-position to the adjacent nitrogen atom may not be substituted, 
 R 3  denotes a C 3-7 -cycloalkyl group, a propargyl group, wherein the unsaturated part may not be linked directly to the nitrogen atom of the R 2 NR 3  group, a phenyl group optionally substituted by a fluorine or chlorine atom, or by a methyl or methoxy group, a pyrazolyl, pyridazolyl or pyridinyl group optionally substituted by a methyl group or 
 R 2  and R 3  together with the nitrogen atom between them denote a 5- to 7-membered cycloalkyleneimino group, optionally substituted by a carboxy or C 1-4 -alkoxycarbonyl group, to which a phenyl ring may additionally be fused, 
   the tautomers, the stereoisomers and the salts thereof.   
     
     
         6 . The antibody molecule of  claim 5 , wherein the anticoagulant is a compound selected from
 (a) 2-[N-(4-amidinophenyl)-aminomethyl]-benzthiazole-5-carboxylic acid-N-phenyl-N-(2-carboxyethyl)-amide,   (b) 2-[N-(4-amidinophenyl)-N-methyl-aminomethyl]-benzthiazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (c) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (d) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(3-hydroxycarbonylpropyl)-amide,   (e) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(hydroxycarbonylmethyl)-amide,   (f) 1-Methyl-2-[2-(2-amidinothiophen-5-yl)ethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (g) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (h) 1-Methyl-2-[2-(4-amidinophenyl)ethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (i) 1-Methyl-2-[2-(4-amidinophenyl)ethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (j) 1-Methyl-2-[2-(4-amidinophenyl)ethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-[2-(1H-tetrazol-5-yl)ethyl]-amide,   (k) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-[2-(1H-tetrazol-5-yl)ethyl]-amide,   (l) 1-Methyl-2-[N-(4-amidinophenyl)-N-methyl-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (m) 1-Methyl-2-[N-(4-amidinophenyl)-N-methyl-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(3-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (n) 1-Methyl-2-[N-(4-amidinophenyl)-N-methyl-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (o) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-[(N-hydroxycarbonylethyl-N-methyl)-2-aminoethyl]-amide,   (p) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(3-fluorophenyl)-N-(2-hydroxycarbonylethyl)-amide,   (q) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(4-fluorophenyl)-N-(2-hydroxycarbonylethyl)-amide,   (r) 1-Methyl-2-[N-(4-amidino-2-methoxy-phenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (s) 1-Methyl-2-[N-(4-amidino-2-methoxy-phenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (t) 1-Methyl-2-[N-(4-amidinophenyl)aminomethyl]-indol-5-yl-carboxylic acid-N-phenyl-N-(2-methoxycarbonylethyl)-amide and   (u) 1-Methyl-2-[N-(4-amidinophenyl)aminomethyl]-thieno[2.3-d]imidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (v) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-phenyl-N-(2-hydroxycarbonylethyl)-amide,   (w) 1-Methyl-2-[N-(4-amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (x) 1-Methyl-2-[N-(4-amidino-2-methoxy-phenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-hydroxycarbonylethyl)-amide,   (y) 1-Methyl-2-[N-[4-(N-n-hexyloxycarbonylamidino)phenyl]-aminomethyl]-benzimidazol-5-yl-carboxylic acid-N-(2-pyridyl)-N-(2-ethoxycarbonylethyl)-amide,   the tautomers, stereoisomers and the salts thereof.   
     
     
         7 . The antibody molecule of  claim 2  wherein the anticoagulant is dabigatran. 
     
     
         8 . The antibody molecule of  claim 7  which is capable of neutralizing the activity of dabigatran and 1-O-acylglucuronide of dabigatran. 
     
     
         9 . The antibody molecule of  claim 5  which comprises a heavy chain variable domain with a CDR1 selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, a CDR2 selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, and a CDR3 selected from the group consisting of SEQ ID NO: 9 and SEQ ID NO: 10, and a light chain variable domain with a CDR1 selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, a CDR2 of SEQ ID NO: 14, and a CDR3 of SEQ ID NO: 15. 
     
     
         10 . The antibody molecule of  claim 9  which comprises a heavy chain variable domain with a CDR1 of SEQ ID NO: 1, a CDR2 selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, a CDR3 of SEQ ID NO: 10, and a light chain variable domain with a CDR1 of SEQ ID NO: 13, a CDR2 of SEQ ID NO: 14, and a CDR3 of SEQ ID NO: 15. 
     
     
         11 . The antibody molecule of  claim 9  which comprises a heavy chain variable domain selected from the group consisting of SEQ ID NOs: 16, 18, 20, 22, 24, and 26, and a light chain variable domain selected from the group consisting of SEQ ID Nos: 17, 19, 21, 23, 25, and 27. 
     
     
         12 . The antibody molecule of  claim 11  which comprises a heavy chain variable domain of SEQ ID NO: 16, and a light chain variable domain of SEQ ID No: 17, or a heavy chain variable domain of SEQ ID NO: 18, and a light chain variable domain of SEQ ID No: 19, or a heavy chain variable domain of SEQ ID NO: 20, and a light chain variable domain of SEQ ID No: 21, or a heavy chain variable domain of SEQ ID NO: 22, and a light chain variable domain of SEQ ID No: 23, or a heavy chain variable domain of SEQ ID NO: 24, and a light chain variable domain of SEQ ID No: 25, or a heavy chain variable domain of SEQ ID NO: 24, and a light chain variable domain of SEQ ID No: 27, or a heavy chain variable domain of SEQ ID NO: 26, and a light chain variable domain of SEQ ID No: 27. 
     
     
         13 . The antibody molecule of  claim 5  which is a polyclonal antibody, a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, a fragment of an antibody, in particular a Fab, Fab′, or F(ab′) 2  fragment, a single chain antibody, in particular a single chain variable fragment (scFv), a Small Modular Immunopharmaceutical (SMIP), a domain antibody, a nanobody, a diabody, or a Designed Ankyrin Repeat Protein (DARPin). 
     
     
         14 . The antibody molecule of  claim 13  which is a scFv, wherein the heavy chain variable domain and the light chain variable domain are linked to each other through a linker peptide selected from the group consisting of SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 31. 
     
     
         15 . The antibody molecule of  claim 14  which comprises SEQ ID NO: 32, or SEQ ID NO: 33. 
     
     
         16 . The antibody molecule of  claim 13  having a heavy chain comprising SEQ ID NO: 34 or SEQ ID NO: 40, and a light chain comprising SEQ ID NO: 35, or having a heavy chain comprising SEQ ID NO: 42, and a light chain comprising SEQ ID NO: 43. 
     
     
         17 . The antibody of  claim 13  which is a Fab molecule having a Fd fragment comprising SEQ ID NO: 36, SEQ ID NO: 38, or SEQ ID NO: 41, and a light chain comprising SEQ ID NO: 37 or SEQ ID NO: 39. 
     
     
         18 . A method for preventing or treating side effects of anticoagulant therapy, or of an overdosing event in anticoagulant therapy, comprising administering an effective amount of an antibody molecule of  claim 5  to a patient in need thereof. 
     
     
         19 . The method according to  claim 18 , wherein the side effect is a bleeding event. 
     
     
         20 . A method of manufacturing an antibody molecule of  claim 5 , comprising
 (a) providing a host cell comprising one or more nucleic acids encoding said antibody molecule in functional association with an expression control sequence,   (b) cultivating said host cell, and   (c) recovering the antibody molecule from the cell culture.   
     
     
         21 . A kit comprising an antibody of  claim 5 , or a pharmaceutical composition thereof. 
     
     
         22 . A kit comprising:
 (a) an antibody of  claim 5 , or a pharmaceutical composition thereof;   (b) a container; and   (c) a label.   
     
     
         23 . A kit comprising an antibody of  claim 7 , and dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof. 
     
     
         24 . The kit according to  claim 23 , wherein the form of dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof is in the form of a solid, liquid or gel. 
     
     
         25 . The kit according to  claim 23 , wherein the pharmaceutically acceptable salt of dabigatran etexilate is a mesylate salt. 
     
     
         26 . The kit according to  claim 23  or  25 , wherein the strength per dosage unit of the dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof is between 75 mg and 300 mg, either QD or BID. 
     
     
         27 . A kit comprising:
 (a) an antibody of  claim 7 , or a pharmaceutical composition thereof;   (b) a pharmaceutical composition of dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof;   (c) a container; and   (d) a label.   
     
     
         28 . The kit according to  claim 27 , wherein the form of dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof is in the form of a solid, liquid or gel. 
     
     
         29 . The kit according to  claim 27 , wherein the pharmaceutically acceptable salt of dabigatran etexilate is a mesylate salt. 
     
     
         30 . The kit according to  claim 27  or  29 , wherein the strength per dosage unit of the dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof is between 75 mg and 300 mg, either QD or BID. 
     
     
         31 . A kit comprising:
 (a) a first pharmaceutical composition comprising dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof;   (b) a second pharmaceutical composition comprising an antibody of  claim 7 ;   (c) instructions for separate administration of said first and second pharmaceutical compositions to a patient,   
       wherein said first and second pharmaceutical compositions are contained in separate containers and said second pharmaceutical composition is administered to a patient requiring neutralization or partial neutralization of dabigatran or 1-O-acylglucuronide of dabigatran. 
     
     
         32 . A method for neutralizing or partially neutralizing dabigatran or 1-O-acylglucuronide of dabigatran in a patient being treated with dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof, comprising administering an antibody of  claim 7 , or a pharmaceutical composition thereof. 
     
     
         33 . A method for neutralizing or partially neutralizing dabigatran or 1-O-acylglucuronide of dabigatran in a patient comprising:
 (a) confirming that a patient was being treated with dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof, and the amount that was taken by the patient;   (b) neutralizing dabigatran or 1-O-acylglucuronide with an antibody of  claim 7  prior to performing a clotting or coagulation test or assay wherein dabigatran or the 1-O-acylglucuronide of dabigatran would interfere with the accurate read out of the test or assay results;   (c) performing the clotting or coagulation test or assay on a sample taken from the patient to determine the level of clot formation without dabigatran or 1-O-acylglucuronide of dabigatran present; and   (d) adjusting an amount of dabigatran, dabigatran etexilate, a prodrug of dabigatran or a pharmaceutically acceptable salt thereof administered to the patient in order to achieve the appropriate balance between clot formation and degradation in a patient.   
     
     
         34 . The method according to  claims 32  or  33 , wherein the amount of antibody to dabigatran or 1-O-acylglucuronide of dabigatran is in the molar ratio of between 0.1 and 100. 
     
     
         35 . The method according to  claim 34 , wherein the amount of antibody to dabigatran or 1-O-acylglucuronide of dabigatran is in the molar ratio of between 0.1 and 10. 
     
     
         36 . The method according to  claim 33 , wherein the accurate read out of the test or assay result is an accurate read out of fibrinogen levels, activated protein C resistance or related tests.