US 2006/0039901 A1 · Antibodies for inhibiting blood coagulation and methods of use thereof

A humanized antibody that binds specifically to human tissue factor (TF) to form a complex, wherein factor X or factor IX binding to the complex and the FX or FIX activation by TF:VIIa are inhibited.

A humanized antibody comprising at least one murine complementarity determining region (CDR), wherein the antibody binds specifically to human tissue factor (TF) to form a complex, and further wherein factor X or factor IX binding to TF or TF:FVIIa and activation by TF:FVIIa thereto is inhibited, the antibody comprising on the heavy chain:

a) a first CDR (CDR1) which is at least 95% identical to CDR1 amino acid sequence shown in FIG. 13B (SEQ ID NO:
8),
b) a second CDR (CDR2) which is at least 95% identical to the CDR2 amino acid sequence shown in FIG. 13C (SEQ ID NOS:
9 or 101),
c) a third CDR (CDR3) which is at least 95% identical to the CDR3 amino acid sequence shown in FIG. 13D (SEQ ID NO:
10),
d) a first framework (FR1) which is at least 95% identical to the FR1 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
e) a second framework (FR2) which is at least 95% identical to the FR2 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
f) a third framework (FR3) which is at least 95% identical to the FR3 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79), and
g) a fourth framework (FR4) which is at least 95% identical to the FR4 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79).

11 more elements

A method of inhibiting blood coagulation in a mammal, the method comprising administering to the mammal, an effective amount of a humanized antibody or fragment thereof wherein the antibody binds specifically to human tissue factor (TF) to form a complex, and further wherein factor X or factor IX binding to TF or TF:FVIIa and activation by TF:FVIIa thereto is inhibited, the antibody or fragment comprising on the heavy chain:

a) a first CDR (CDR1) which is at least 95% identical to CDR1 amino acid sequence shown in FIG. 13B (SEQ ID NO:
8),
b) a second CDR (CDR2) which is at least 95% identical to the CDR2 amino acid sequence shown in FIG. 13C (SEQ ID NOS:
9 or 101),
c) a third CDR (CDR3) which is at least 95% identical to the CDR3 amino acid sequence shown in FIG. 13D (SEQ ID NO:
10),
d) a first framework (FR1) which is at least 95% identical to the FR1 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
e) a second framework (FR2) which is at least 95% identical to the FR2 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
f) a third framework (FR3) which is at least 95% identical to the FR3 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
g) a fourth framework (FR4) which is at least 95% identical to the FR4 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91);
and on the light chain,
h) a first CDR (CDR1) which is at least 95% identical to CDR1 amino acid sequence shown in FIG. 12B (SEQ ID NO:
2),
i) a second CDR (CDR2) which is at least 95% identical to the CDR2 amino acid sequence shown in FIG. 12C (SEQ ID NO:
6),
j) a third CDR (CDR3) which is at least 95% identical to the CDR3 amino acid sequence shown in FIG. 12D (SEQ ID NO:
7),
k) a first framework (FR1) which is at least 95% identical to the FR1 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79,
l) a second framework (FR2) which is at least 95% identical to the FR2 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
m) a third framework (FR3) which is at least 95% identical to the FR3 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
n) a fourth framework (FR4) which is at least 95% identical to the FR4 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
o) a light chain constant region which is at least 95% identical to the amino acid sequence shown in FIG. 14A (SEQ ID NO:
97) or FIG. 15A (SEQ ID NO:
99), and
p) a heavy chain constant region which is at least 95% identical to the amino acid sequence shown in FIG. 14B (SEQ ID NO:
98) or FIG. 15B (SEQ ID NO:
100).

32 more elements

A method of inhibiting blood coagulation in a mammal, the method comprising administering to the mammal, an effective amount of a humanized antibody or fragment thereof wherein the antibody binds specifically to human tissue factor (TF) to form a complex, and further wherein factor X or factor IX binding to TF or TF:FVIIa and activation by TF:FVIIa thereto is inhibited, the antibody or fragment comprising on the heavy chain:

a) a first CDR (CDR1) identical to CDR1 amino acid sequence shown in FIG. 13B (SEQ ID NO:
8),
b) a second CDR (CDR2) identical to the CDR2 amino acid sequence shown in FIG. 13C (SEQ ID NOS:
9 or 101),
c) a third CDR (CDR3) identical to the CDR3 amino acid sequence shown in FIG. 13D (SEQ ID NO:
10),
d) a first framework (FR1) identical to the FR1 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
e) a second framework (FR2) identical to the FR2 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
f) a third framework (FR3) identical to the FR3 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91),
g) a fourth framework (FR4) identical to the FR4 amino acid sequence shown in FIG. 13A (SEQ ID NO:
91);
and on the light chain:
h) a first CDR (CDR1) identical to CDR1 amino acid sequence shown in FIG. 12B (SEQ ID NO:
2),
i) a second CDR (CDR2) identical to the CDR2 amino acid sequence shown in FIG. 12C (SEQ ID NO:
6),
j) a third CDR (CDR3) identical to the CDR3 amino acid sequence shown in FIG. 12D (SEQ ID NO:
7),
k) a first framework (FR1) identical to the FR1 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
l) a second framework (FR2) identical to the FR2 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
m) a third framework (FR3) identical to the FR3 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
n) a fourth framework (FR4) identical to the FR4 amino acid sequence shown in FIG. 12A (SEQ ID NO:
79),
o) a light chain constant region which is identical to the amino acid sequence shown in FIG. 14A (SEQ ID NO:
97) or FIG. 15A (SEQ ID NO:
99), and p) a heavy chain constant region which is identical to the amino acid sequence shown in FIG. 14B (SEQ ID NO:
98) or FIG. 15B (SEQ ID NO:
100).

31 more elements

A method for producing a humanized antibody, wherein the method comprises:

a) comparing the amino acid sequence of a light chain framework from a rodent antibody against a collection of corresponding human antibody framework sequences,
b) selecting a human framework sequence from the collection having the greatest amino acid sequence identity to the corresponding rodent light chain framework,
c) mutagenizing a DNA segment encoding the rodent light chain framework to encode a humanized light chain framework having an amino acid sequence that is substantially identical to the human framework sequence selected in step b),
d) repeating steps a) thru c) for each individual framework of the rodent light chain to produce a plurality of DNA sequences in which each sequence encodes a humanized light chain framework, wherein each of the corresponding human framework sequences selected in step b) are from the same or different human antibody,
e) assembling into a first vector encoding at least the light chain variable region of the rodent antibody, the DNA sequences encoding the humanized framework sequences produced in step d);
and
f) introducing the assembled vector into a host under conditions sufficient to produce the humanized antibody.

4 more elements

Antibodies for inhibiting blood coagulation and methods of use thereof

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Antibodies for inhibiting blood coagulation and methods of use thereof

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Abstract

Claims72