Monoclonal antibody

• US 20090017040A1
• Filed: 06/12/2008
• Published: 01/15/2009
• Est. Priority Date: 06/12/2007
• Status: Active Grant

• Current Assignee: AC Immune SA, Genentech, Inc. (Roche Holding AG)
• Original Assignee: AC Immune SA, Genentech, Inc. (Roche Holding AG)
• Inventors: Pihlgren, Maria, Muhs, Andreas, Watts, Ryan, Pfeifer, Andrea
• Granted Patent: US 8,613,923 B2
• Time in Patent Office: Days
• US Class Current: 424/152.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 31/5513 1,4-Benzodiazepines, e.g. diazepam or clozapine
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: G01N 2333/4709 Amyloid plaque core protein
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention is related to methods and compositions for the therapeutic and diagnostic use in the treatment of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of disorders and abnormalities associated with amyloid protein such as Alzheimer'"'"'s disease. The present invention provides novel methods and compositions comprising highly specific and highly effective antibodies having the ability to specifically recognize and bind to specific epitopes from a range of β-amyloid proteins. The antibodies enabled by the teaching of the present invention are particularly useful for the treatment of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis including, but not limited to, neurological disorders such as Alzheimer'"'"'s Disease (AD)

Claims

1-92. -92. (canceled)

93. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody recognizes and binds to a conformational epitope and binds to polymeric soluble amyloid, amyloid fibrils, amyloid fibers, polymeric soluble Aβ

peptides comprising a plurality of Aβ

_1-42 monomeric peptides and/or Aβ

fibrils or fibers incorporating a plurality of said polymeric peptides, respectively.

94. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 93, which antibody binds to Aβ

monomeric peptide 1-42, wherein the antibody has essentially no binding to Aβ

monomeric peptide 17-40 and substantially weaker binding to Aβ

monomeric peptide 1-28.

95. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 93, which antibody, upon co-incubation with an Aβ

peptide comprising a number of amino acid residues selected from at least 30, at least 35, at least 38, at least 40 amino acid residues, and 42 amino acid residues in a monomeric and/or an oligomeric form inhibits the aggregation of the Aβ

monomers and/or oligomers to high molecular weight polymeric fibrils.

96. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 93, wherein the antibody binds to Aβ

1-42 monomeric peptide and also to polymeric soluble Aβ

peptides comprising a plurality of Aβ

1-42 monomeric peptides and Aβ

fibrils or fibers incorporating a plurality of said polymeric peptides, but shows a substantially weaker binding to Aβ

monomeric peptide 1-28 and/or essentially no binding to Aβ

monomeric peptide 17-40 and, upon co-incubation with preformed high molecular weight polymeric amyloid fibrils or filaments formed by the aggregation of Aβ

_1-42 monomeric and/or oligomeric peptides is capable of disaggregating the preformed polymeric fibrils or filaments, by at least 10%, by at least 20%, particularly by at least 30%, more particularly by at least 40%, even more particularly by at least 50%, but especially by at least 60%, by at least 70%, by at least 80% or more.

97. A monoclonal antibody according to claim 93 including any functionally equivalent antibody or functional parts thereof, which antibody exhibits high specificity to Aβ

_1-42 monomeric peptides and to polymeric soluble Aβ

peptides comprising a plurality of Aβ

1-42 monomeric peptides and Aβ

fibrils or fibers incorporating a plurality of said polymeric peptides, but shows essentially no or moderate cross-reactivity to Aβ

_1-28 and/or Aβ

_17-40 monomeric peptides.

98. A monoclonal antibody according to claim 93 including any functionally equivalent antibody or functional parts thereof which antibody is capable of decreasing the total amount of soluble Aβ

in the brain of a subject suffering from a disease or condition associated with increased concentrations of soluble Aβ

in the brain.

99. A monoclonal antibody according to claim 93 including any functionally equivalent antibody or functional parts thereof which antibody is capable of solubilizing and/or disrupting plaques thus decreasing the plaque load in the brain of an animal, particularly a mammal, but especially a human suffering from a disease or condition associated with an increased plaque load in the brain.

116. An isolated polynucleotide encoding the antibody or functional part thereof of claim 93.

119. A therapeutic composition comprising the monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 93 in a therapeutically effective amount, and optionally further comprising a pharmaceutically acceptable carrier, a diluent and/or an excipient.

120. A method for treating diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, comprising administration of the antibody or functional part thereof of claim 93.

121. The composition of claim 119, further comprising a further biologically active substance selected from one or more compounds used in the medication of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, compounds against oxidative stress, anti-apoptotic compounds, metal chelators, inhibitors of DNA repair such as pirenzepin and metabolites, 3-amino-1-propanesulfonic acid (3APS), 1,3-propanedisulfonate (1,3PDS), secretase activators, β

- and γ

-secretase inhibitors, tau proteins, neurotransmitter, β

-sheet breakers, anti-inflammatory molecules, or cholinesterase inhibitors (ChEIs) such as tacrine, rivastigmine, donepezil, galantamine and/or nutritive supplements.

122. A method for producing an antibody including any functionally equivalent antibody or functional parts thereof according to claim 93, which method comprises raising in a suitable host organism antibodies against a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of a β

-amyloid peptide selected from Aβ

_22-35 and Aβ

_29-40, respectively, modified with one or more hydrophilic moieties, or modified with a hydrophilic polyethylene glycol (PEG) moiety, wherein said hydrophilic moiety is covalently bound to each terminus through an amino acid selected from lysine, glutamic acid, cysteine, and any other suitable amino acid or amino acid analogue capable of serving as a linker molecule; and

isolating the antibody.

123. A method of treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, neurological disorders such as Alzheimer'"'"'s Disease (AD and diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), Lewy body dementia, Down'"'"'s syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type);

the Guam Parkinson-Dementia complex; and

other diseases which are based on or associated with amyloid-like proteins such as progressive supranuclear palsy, multiple sclerosis;

Creutzfeld Jacob disease, Parkinson'"'"'s disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), inclusion-body myositis (IBM), Adult Onset Diabetes;

senile cardiac amyloidosis;

endocrine tumors, and macular degeneration in a subject, comprising administration of a monoclonal antibody or functional part thereof according to claim 93 to the subject such that the effects of the disease or disorder are treated and/or alleviated.

124. A method for treating or alleviating the effects of diseases and disorders which are caused by or associated with amyloid or amyloid-like proteins including amyloidosis, a group of diseases and disorders associated with amyloid plaque formation including secondary amyloidosis and age-related amyloidosis such as diseases including, but not limited to, neurological disorders such as Alzheimer'"'"'s Disease (AD) and diseases or conditions characterized by a loss of cognitive memory capacity such as, for example, mild cognitive impairment (MCI), Lewy body dementia, Down'"'"'s syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type);

the Guam Parkinson-Dementia complex;

as well as other diseases which are based on or associated with amyloid-like proteins such as progressive supranuclear palsy, multiple sclerosis;

Creutzfeld Jacob disease, Parkinson'"'"'s disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), Adult Onset Diabetes;

senile cardiac amyloidosis;

endocrine tumors, and others, including macular degeneration, comprising administration of the composition of claim 119.

126. A method for reducing the plaque load in the brain of a subject suffering from a disease or condition associated with an increased plaque load in the brain comprising administering to the subject a therapeutically effective amount of a monoclonal antibody according to claim 93.

127. A method for reducing the amount of plaques in the brain of a subject suffering from a disease or condition associated with an increased plaque load in the brain comprising administering to the subject a therapeutically effective amount of a monoclonal antibody according to claim 93.

128. A method for decreasing the total amount of soluble Aβ

in the brain of a subject suffering from a disease or condition associated with increased concentrations of soluble Aβ

in the brain comprising administering to the subject a therapeutically effective amount of a monoclonal antibody according to claim 93.

129. A method for retaining or increasing cognitive memory capacity in a subject exhibiting an amyloid-associated disease or condition comprising administering to the subject a therapeutically effective amount of a monoclonal antibody of claim 93.

130. A cell line characterized in that it produces a monoclonal antibody of claim 93.

133. A method of diagnosis of or of diagnosing a predisposition to an amyloid-associated disease or condition in a patient comprising detecting the immunospecific binding of a monoclonal antibody or an active fragment thereof to an epitope of the amyloid protein in a sample or in situ which includes the steps of

(a) bringing the sample or a specific body part or body area suspected to contain the amyloid protein into contact with a monoclonal antibody according to claim 93, which antibody binds an conformational epitope of the amyloid protein;

(b) allowing the antibody to bind to the amyloid protein to form an immunological complex;

(c) detecting the formation of the immunological complex; and

(d) correlating the presence or absence of the immunological complex with the presence or absence of amyloid protein in the sample or specific body part or area.

134. A method of determining the extent of amyloidogenic plaque burden in a tissue comprising

(a) obtaining a sample representative of the tissue under investigation;

(b) testing said sample for the presence of amyloid protein with a monoclonal antibody according to claim 93;

(c) determining the amount of antibody bound to the antigen; and

(d) calculating the plaque burden in the tissue.

135. A method for monitoring minimal residual disease in a patient following treatment with at least one monoclonal antibody or functional part thereof of claim 93, wherein said method comprises:

(a) bringing the sample or a specific body part or body area suspected to contain the amyloid protein into contact with a monoclonal antibody according to claim 93, which antibody binds an conformational epitope of the amyloid protein;

(b) allowing the antibody to bind to the amyloid protein to form an immunological complex;

(c) detecting the formation of the immunological complex;

(d) correlating the presence or absence of the immunological complex with the presence or absence of amyloid protein in the sample or specific body part or area; and

(e) comparing the amount of said immunological complex to a normal control value,

wherein an increase in the amount of said aggregate compared to a normal control value indicates that said patient still suffers from a minimal residual disease.

136. A method for predicting responsiveness of a patient being treated with at least one monoclonal antibody or functional part thereof of claim 193, comprising

(a) bringing the sample or a specific body part or body area suspected to contain the amyloid protein into contact with a monoclonal antibody according to claim 93, which antibody binds an conformational epitope of the amyloid protein;

(b) allowing the antibody to bind to the amyloid protein to form an immunological complex;

(c) detecting the formation of the immunological complex;

(d) correlating the presence or absence of the immunological complex with the presence or absence of amyloid protein in the sample or specific body part or area; and

(e) comparing the amount of said immunological complex before and after onset of the treatment,

wherein a decrease in the amount of said aggregate indicates that said patient has a high potential of being responsive to the treatment.

137. A test kit comprising at least one antibody or functional part thereof of claim 93, and optionally further comprising instructions for using the antibodies for the purpose of binding to amyloid protein to form an immunological complex and detecting the formation of the immunological complex such that presence or absence of the immunological complex correlates with presence or absence of amyloid protein.

141. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 93, which antibody binds to Aβ

monomeric peptides having at least 30, particularly at least 35, more particularly at least 38, even more particularly at least 40 amino acid residues but shows essentially no binding to Aβ

monomeric peptide 17-40.

142. A polynucleotide which hybridizes under stringent conditions to the polynucleotide sequence of claim 116.

100. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma cell line FG1F9E4, deposited on May 25, 2007 and given deposit number DSM ACC2845.

101. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FG1F9E4, deposited on May 25, 2007 and given deposit number DSM ACC2845.

102. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma cell line FK2A6A6, deposited on May 25, 2007 and given deposit number DSM ACC2846.

103. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof produced by hybridoma cell line FK2A6A6, deposited on May 25, 2007 and given deposit number DSM ACC2846.

104. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody has been raised against a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of the β

-amyloid peptide, Aβ

_22-35 and Aβ

_29-40, respectively, modified with a hydrophilic polyethylene glycol (PEG) moiety, wherein said hydrophilic moiety is covalently bound to each terminus through an amino acid selected from lysine, glutamic acid, cysteine, and any other suitable amino acid or amino acid analogue capable of serving as a linker molecule.

125. A therapeutic composition comprising the monoclonal antibody including any functionally equivalent antibody or functional parts thereof according to claim 104 in a therapeutically effective amount, and optionally further comprising a pharmaceutically acceptable carrier, a diluent, and/or an excipient.

105. A light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a sequence selected from SEQ ID NO:

7 and SEQ ID NO;

9, or a functional part thereof comprising at least one, at least two, or at least 3 of the light chain CDR having a polypeptide sequence selected from one or more of SEQ ID NOs;

11-13.

112. An isolated polynucleotide encoding the light chain variable region of claim 105.

106. A heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a sequence selected from SEQ ID NO:

8 and SEQ ID NO;

10, or a functional part thereof comprising at least one, at least two, or at least 3 of the heavy chain CDR having a polypeptide sequence selected from one or more of SEQ ID NOs;

14-16.

113. An isolated polynucleotide encoding the heavy chain variable region of claim 106.

107. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody comprises a light chain variable domain comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a sequence selected from SEQ ID NO:

7 and SEQ ID NO;

9, or a functional part thereof comprising at least one, at least two, or at least 3 of the light chain CDRs having a polypeptide sequence selected from one or more of SEQ ID NOs;

11-13.

114. An isolated polynucleotide encoding the antibody or functional parts thereof according to claim 107.

108. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody comprises a heavy chain variable domain comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a sequence selected from SEQ ID NO:

8 and SEQ ID NO;

10, or a functional part thereof comprising at least one, at least two, or at least 3 of the heavy chain CDRs having a polypeptide sequence selected from one or more of SEQ ID NOs;

14-16.

115. An isolated polynucleotide encoding the antibody or functional parts thereof according to claim 108.

109. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof comprising the amino acid sequences of SEQ ID NOs:

7-8, wherein the antibody has been altered by introducing at least one, at least two, or at least 3 or more conservative substitutions into the sequences of SEQ ID NOs;

7-8, wherein the antibody essentially maintains its full functionality, and wherein the amino acid in position 52 is selected from any amino acid, from the group consisting of cysteine, tyrosine, and serine, and from cysteine alone.

139. An Aβ

epitope which is specifically recognized by the monoclonal antibody or functional part thereof of claim 109.

110. A monoclonal antibody including any functionally equivalent antibody or functional parts thereof comprising the amino acid sequences of SEQ ID NOs:

9-10, wherein the antibody has been altered by introducing at least one, at least two, or at least 3 or more conservative substitutions into the sequences of SEQ ID NOs;

9-10, wherein the antibody essentially maintains its full functionality, and wherein the amino acid in position 52 is selected from any amino acid, from the group consisting of cysteine, tyrosine, and serine, and from cysteine alone.

140. An Aβ

epitope which is specifically recognized by the monoclonal antibody or functional part thereof of claim 110.

111. An isolated CDR comprising an amino acid sequence selected from SEQ ID NOs:

11-13.

117. An isolated polynucleotide comprising at least one of the nucleotide sequences set forth in SEQ ID NOs:

17-18.

143. A polynucleotide which hybridizes under stringent conditions to the polynucleotide sequence of claim 117.

118. An isolated polynucleotide comprising at least one of the nucleotide sequences set forth in SEQ ID NOs:

19-20.

144. A polynucleotide which hybridizes under stringent conditions to the polynucleotide sequence of claim 118.

131. A cell line characterized in that it produces a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma FG1F9E4, deposited on May 25, 2007 and given deposit number DSM ACC2845, or wherein the cell line is hybridoma FG1F9E4, deposited on May 25, 2007 and given deposit number DSM ACC2845.

132. A cell line characterized in that it produces a monoclonal antibody including any functionally equivalent antibody or functional parts thereof which antibody has the characteristic properties of an antibody produced by hybridoma FK2A6A6, deposited on May 25, 2007 and given deposit number DSM ACC2846, or wherein the cell line is hybridoma FK2A6A6, deposited on May 25, 2007 and given deposit number DSM ACC2846.

138. An Aβ

epitope which is specifically recognized by a monoclonal antibody including any functionally equivalent antibody or functional parts thereof, which antibody has been raised against a supramolecular antigenic construct comprising an antigenic peptide corresponding to the amino acid sequence of the β

-amyloid peptide, Aβ

_22-35 and Aβ

_29-40, respectively, modified with a hydrophilic polyethylene glycol (PEG) moiety, wherein said hydrophilic moiety is covalently bound to each terminus through an amino acid such as, for example, lysine or any other suitable amino acid or amino acid analogue capable of serving as a linker molecule.