ANTAGONISTS OF PCSK9
Antagonists of human proprotein convertase subtilisin-kexin type 9 (“PCSK9”) are disclosed. The disclosed antagonists are effective in the inhibition of PCSK9 function and, accordingly, present desirable antagonists for the use in the treatment of conditions associated with PCSK9 activity. The present invention also discloses nucleic acid encoding said antagonists, vectors, host cells, and compositions comprising the antagonists. Methods of making PCSK9-specific antagonists as well as methods of using the antagonists for inhibiting or antagonizing PCSK9 function are also disclosed and form important additional aspects of the present disclosure.
1 . An isolated PCSK9-specific antagonist that antagonizes PCSK9's inhibition of cellular LDL uptake and comprises:
(a) a heavy chain variable region comprising a CDR3 domain comprising SEQ ID NO: 67 or an equivalent thereof characterized as having one or more conservative amino acid substitutions in the CDR3 domain; and/or
(b) a light chain variable region comprising a CDR3 domain comprising SEQ ID NO: 57 or an equivalent thereof characterized as having one or more conservative amino acid substitutions in the CDR3 domain.
2 . The PCSK9-specific antagonist of claim 1 that binds to human PCSK9 with an equilibrium dissociation constant (KD) of less than 1200 nM.
3 . The PCSK9-specific antagonist of claim 1 that binds to human PCSK9 with a KD of less than 500 nM.
4 . The PCSK9-specific antagonist of claim 1 that binds to human PCSK9 with a KD of less than 100 nM.
5 . The PCSK9-specific antagonist of claim 1 that binds to human PCSK9 with a KD of less than 5 nM.
6 . The PCSK9-specific antagonist of claim 1 that antagonizes PCSK9's inhibition of cellular LDL uptake at an IC50 of less than 500 nM.
7 . The PCSK9-specific antagonist of claim 1 that antagonizes PCSK9's inhibition of cellular LDL uptake at an IC50 of less than 200 nM.
8 . The PCSK9-specific antagonist of claim 1 that antagonizes PCSK9's inhibition of cellular LDL uptake at an IC50 of less than 100 nM.
9 . The PCSK9-specific antagonist of claim 1 which is an antibody molecule.
10 . The PCSK9-specific antagonist of claim 1 which comprises:
(a) a heavy chain variable CDR1 sequence comprising SEQ ID NO: 63;
(b) a heavy chain variable CDR2 sequence comprising SEQ ID NO: 65;
(c) a light chain variable CDR1 sequence comprising SEQ ID NO:55; and/or
(d) a light chain variable CDR2 sequence comprising SEQ ID NO: 39.
11 . The PCSK9-specific antagonist of claim 1 which comprises a heavy chain variable region comprising SEQ ID NO: 61 and/or a light chain variable region comprising SEQ ID NO: 53.
12 . The PCSK9-specific antagonist of claim 9 which comprises a heavy chain comprising constant sequence comprising: SEQ ID NO: 87.
13 . A composition comprising the PCSK9-specific antagonist of claim 1 and a pharmaceutically acceptable carrier.
14 . A method for antagonizing PCSK9 function which comprises employing a PCSK9-specific antagonist of claim 1 .
15 . (canceled)
16 . Isolated nucleic acid encoding a PCSK9-specific antagonist of claim 1 .
17 . Isolated nucleic acid which encodes a PCSK9-specific antagonist of claim 1 which comprises:
(a) a heavy chain variable region wherein the CDR3 domain is encoded by nucleic acid sequence comprising SEQ ID NO: 68; and/or
(b) a light chain variable region wherein the CDR3 domain is encoded by nucleic acid sequence comprising SEQ ID NO: 58.
18 . The isolated nucleic acid of claim 17 which encodes an antibody molecule which comprises:
(a) a heavy chain variable region; said heavy chain variable region which comprises CDR1 and/or CDR2 domains, respectively, encoded by nucleic acid sequence comprising at least one nucleic acid sequence selected from the group consisting of: SEQ ID NO: 64 and SEQ ID NO: 66; and/or
(b) a light chain variable region; said light chain variable region which comprises CDR1 and/or CDR2 domains, respectively, encoded by nucleic acid sequence comprising at least one nucleic acid sequence selected from the group consisting of: SEQ ID NO: 56 and SEQ ID NO: 40.
19 . The isolated nucleic acid of claim 17 which encodes an antibody molecule which comprises:
(a) a heavy chain variable region wherein the heavy chain variable region is encoded by nucleic acid sequence comprising SEQ ID NO: 62; and/or
(b) a light chain variable region wherein the light chain variable region is encoded by nucleic acid sequence comprising SEQ ID NO: 54.
20 . A vector comprising nucleic acid of claim 16 .
21 . An isolated host cell or population of host cells in vitro or in situ comprising nucleic acid of claim 16 .
22 . A method for producing a PCSK9-specific antagonist which comprises:
(a) culturing the cell(s) of claim 21 under conditions appropriate for production of the PCSK9-specific antagonist; and
(b) isolating the PCSK9-specific antagonist produced.
23 . An isolated host cell or population of host cells in vitro or in situ comprising a PCSK9-specific antagonist of claim 1 .