The invention provides methods and compositions for modulating hepatocyte growth factor activator function.

Description of the Invention

The invention provides methods, compositions, kits and articles of manufacture for modulating hepatocyte growth factor activator (HGFA) function, thereby modulating physiological effects of HGFA activity. Modulation of HGFA function can be effected by the use of antibodies as described herein.

The invention provides modulator molecules capable of use for modulating HGFA function. In one embodiment, HGFA function is modulated through inhibition of HGFA activity (e.g., proteolytic activity). Generally, the modulator molecules comprise an antibody as described herein. The modulator molecules are capable of effecting modulation either directly (e.g., by binding to HGFA and interefering with HGFA proteolytic activity) or indirectly (e.g., by targeting/directing an active agent to HGFA in a tissue or cell, wherein the active agent is capable of interfering with HGFA proteolytic activity). In one embodiment, the invention provides an antagonist molecule comprising an antibody that binds to HGFA. In one embodiment, binding of the antagonist to HGFA interferes with HGFA proteolytic activity. In one embodiment, binding of the antagonist to HGFA interferes with activation of HGF by HGFA. In one embodiment, the antibody binds to the active site of HGFA. In one embodiment, the antibody binds to HGFA at a position other than the HGFA active site (e.g., an exosite). In one embodiment, binding of the antibody to HGFA at a position other than the HGFA active site inhibits interaction of HGFA with its substrate molecule. In one embodiment, binding of the antibody to HGFA at a position other than the HGFA active site inhibits HGFA proteolytic activity.

In one aspect, the invention provides antagonists that disrupt the HGF/c-met signaling pathway. For example, the invention provides a molecule that inhibits HGFA cleavage of proHGF (e.g., cleavage at the R494-V495 position). The molecule can exert its inhibitory function in any number of ways, including but not limited to binding to HGFA at its active site and/or at a site other than the active site (e.g., an exosite) such that HGFA cleavage of proHGF is inhibited. The molecule can bind to HGFA in complexed or uncomplexed form. The molecule can also exert its inhibitory function by interfering with one or more aspects of the HGF activation process. For example, in one embodiment, an antagonist molecule of the invention binds to HGFA-proHGF complex such that cleavage of proHGF is inhibited. In one embodiment, binding of the molecule to proHGF or HGFA (singly or in complex) inhibits release of HGF subsequent to cleavage by HGFA. In one embodiment, an antagonist molecule of the invention does not inhibit HGF binding to c-met. For example, in one embodiment, an antagonist molecule of the invention is not an antibody or fragment thereof having similar inhibitory and/or binding ability as the antibody produced by hybridoma cell line deposited under American Type Culture Collection Accession Number ATCC HB-11894 (hybridoma 1A3.3.13) or HB-11895 (hybridoma 5D5.11.6). In one embodiment, an antagonist molecule of the invention inhibits biological activities associated with HGF/c-met activation.

In one aspect, the invention provides an antibody comprising a CDR-H1 region comprising the sequence of SEQ ID NO:3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39 or 42. In one aspect, the invention provides an antibody comprising a CDR-H2 region comprising the sequence of SEQ ID NO:4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40 or 43. In one aspect, the invention provides an antibody comprising a CDR-H3 region comprising the sequence of SEQ ID NO:5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41 or 44. In one embodiment, the invention provides an antibody comprising a CDR-H1 region comprising the sequence of SEQ ID NO:3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39 or 42, and a CDR-H2 region comprising the sequence of SEQ ID NO:4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40 or 43. In one embodiment, the invention provides an antibody comprising a CDR-H1 region comprising the sequence of SEQ ID NO:3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39 or 42, and a CDR-H3 region comprising the sequence of SEQ ID NO:5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41 or 44. In one embodiment, the invention provides an antibody comprising a CDR-H2 region comprising the sequence of SEQ ID NO:4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40 or 43, and a CDR-H3 region comprising the sequence of SEQ ID NO:5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41 or 44. In one embodiment, the invention provides an antibody comprising a CDR-H1 region comprising the sequence of SEQ ID NO:3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39 or 42, a CDR-H2 region comprising the sequence of SEQ ID NO:4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40 or 43, and a CDR-H3 region comprising the sequence of SEQ ID NO:5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41 or 44.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:3;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:4;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:5.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:6;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:7;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:8.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:9;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:10;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:11.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:12;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:13;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:14.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:15;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:16;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:17.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:18;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:19;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:20.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:21;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:22;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:23.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:24;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:25;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:26.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:27;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:28;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:29.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:30;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:31;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:32.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:33;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:34;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:35.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:36;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:37;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:38.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:39;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:40;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:41.

In one aspect, the invention provides an antibody comprising at least one, at least two, or all three of the following:

(i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:42;

(ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:43;

(iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:44.

The amino acid sequences of SEQ ID NOs:3-44 are numbered with respect to individual CDR (i.e., H1, H2 or H3) as indicated in FIG. 1 (see Original Patent), the numbering being consistent with the Kabat numbering system as described below.

In one embodiment, an antibody of the invention comprises a heavy chain variable domain CDR sequence(s) comprising the sequence of at least one, at least two, or all three of the H1 (SEQ ID NO: 71-84), H2 (SEQ ID NO: 85-98) and/or H3 (SEQ ID NO: 99-112) sequences for each clone depicted in FIGS. 1B, 1C and 1D (see Original Patent).

In one aspect, the invention provides antibodies comprising heavy chain CDR sequences as depicted in FIGS. 1A, B, C and D. In some embodiment, these antibodies further comprise a light chain variable domain of humanized 4D5 antibody (huMAb4D5-8) (HERCEPTIN.RTM., Genentech, Inc., South San Francisco, Calif., USA) (also referred to in U.S. Pat. No. 6,407,213 and Lee et al., J. Mol. Biol. (2004), 340(5):1073-93) as depicted in SEQ ID NO:45 below -- see Original Patent.

In one embodiment, the huMAb4D5-8 light chain variable domain sequence is modified at one or more of positions 30, 66 and 91 (Asn, Arg and His as indicated in bold/italics above, respectively). In one embodiment, the modified huMAb4D5-8 sequence comprises Ser in position 30, Gly in position 66 and/or Ser in position 91. Accordingly, in one embodiment, an antibody of the invention comprises a light chain variable domain comprising the sequence depicted in SEQ ID NO: 54 below -- see Original Patent.

Substituted residues with respect to huMAb4D5-8 are indicated in bold/italics above.

Antibodies of the invention can further comprise any suitable framework and/or light chain variable domain sequences, provided HGFA binding activity is substantially retained. For example, in some embodiments, these antibodies further comprise a human subgroup III heavy chain framework consensus sequence. In one embodiment of these antibodies, the framework consensus sequence comprises substitution at position 71, 73 and/or 78. In some embodiments of these antibodies, position 71 is A, 73 is T and/or 78 is A. In one embodiment, these antibodies comprise heavy chain variable domain framework sequences of humanized 4D5 antibody (huMAb 4D5-8) (HERCEPTIN.RTM., Genentech, Inc., South San Francisco, Calif., USA) (also referred to in U.S. Pat. No. 6,407,213 and Lee et al., J. Mol. Biol. (2004), 340(5):1073-93). In one embodiment, the humanized 4D5-8 antibody is as described in U.S. Pat. No. 6,407,213. In one embodiment, these antibodies further comprise a human .kappa.I light chain framework consensus sequence. In one embodiment, these antibodies comprise light chain variable domain sequences of humanized 4D5 antibody (huMAb 4D5-8) (SEQ ID NO:45) (HERCEPTIN.RTM., Genentech, Inc., South San Francisco, Calif., USA) (also referred to in U.S. Pat. No. 6,407,213 and Lee et al., J. Mol. Biol. (2004), 340(5):1073-93), or the modified variant thereof as depicted in SEQ ID NO: 54.

In one embodiment, an antibody of the invention comprises a heavy chain variable domain, wherein the framework sequence comprises the sequences of SEQ ID NO: 46, 47, 48 and 49 (FR1, 2, 3, and 4, respectively), and CDR H1, H2 and H3 sequences as depicted in FIGS. 1A, B, C, and/or D. In one embodiment, an antibody of the invention comprises a light chain variable domain, wherein the framework sequence comprises the sequence of SEQ ID NO: 50, 51, 52 and 53 (FR1, 2, 3, and 4, respectively), and CDR L1, L2 and L3 sequences as depicted in SEQ ID NO: 54.

In one embodiment, an antibody of the invention comprises a heavy chain variable domain, wherein the framework sequence comprises the sequence of SEQ ID NO: 59, 60, 61 and 62 (FR1, 2, 3 and 4, respectively) (FIG. 1E (see Original Patent)), and CDR H1, H2 and H3 sequences as depicted in FIG. 1. In one embodiment, an antibody of the invention comprises a light chain variable domain, wherein the framework sequence comprises the sequence of SEQ ID NO: 55, 56, 57, and 58 (FR 1, 2, 3 and 4, respectively) (FIG. 1E), and CDR L1, L2 and L3 sequences as depicted in SEQ ID NO: 54.

In one embodiment, an antibody of the invention comprises a heavy chain variable domain, wherein the framework sequence comprises the sequence of SEQ ID NO: 67, 68, 69 and 70 (FR 1, 2, 3 and 4, respectively) (FIG. 1F (see Original Patent)), and CDR H1, H2 and H3 sequences as depicted in FIGS. 1A, B, C and/or D (see Original Patent). In one embodiment, an antibody of the invention comprises a light chain variable domain, wherein the framework sequence comprises the sequence of SEQ ID NO: 63, 64, 65, and 66 (FR 1, 2, 3 and 4, respectively) (FIG. 1F), and CDR L1, L2 and L3 sequences as depicted in SEQ ID NO: 54.

In one aspect, the invention provides an antibody that competes with any of the above-mentioned antibodies for binding to HGFA. In one aspect, the invention provides an antibody that binds to the same epitope on HGFA as any of the above-mentioned antibodies. In one embodiment, an antibody of the invention is affinity matured, humanized, chimeric, or human. In one embodiment, an antibody of the invention is an antibody fragment (as described herein), or a substantially full length antibody. In one embodiment, an antibody of the invention comprises a wild type Fc region, or a variant thereof. In one embodiment, an antibody of the invention is an IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgM, IgE or IgD.

In one aspect, an antagonist molecule of the invention is linked to a toxin such as a cytotoxic agent. These molecules/substances can be formulated or administered in combination with an additive/enhancing agent, such as a radiation and/or chemotherapeutic agent.

The invention also provides methods and compositions useful for modulating disease states associated with dysregulation of the HGF/c-met signaling axis. Thus, in one aspect, the invention provides a method of modulating c-met activation in a subject, said method comprising administering to the subject a modulator molecule of the invention that inhibits HGFA cleavage of proHGF, whereby c-met activation is modulated. In one aspect, the invention provides a method of treating a pathological condition associated with activation of c-met in a subject, said method comprising administering to the subject a modulator molecule of the invention that inhibits HGFA cleavage of proHGF, whereby c-met activation is inhibited. In one embodiment, the modulator molecule of the invention is an antibody that binds to HGFA.

The HGF/c-met signaling pathway is involved in multiple biological and physiological functions, including, e.g., cell growth stimulation (e.g. cell proliferation, cell survival, cell migration, cell morphogenesis) and angiogenesis. Thus, in another aspect, the invention provides a method of inhibiting c-met activated cell growth (e.g. proliferation and/or survival), said method comprising contacting a cell or tissue with an antagonist of the invention, whereby cell proliferation associated with c-met activation is inhibited. In yet another aspect, the invention provides a method of inhibiting angiogenesis, said method comprising administering to a cell, tissue, and/or subject with a condition associated with abnormal angiogenesis an antagonist of the invention, whereby angiogenesis is inhibited.

In one aspect, the invention provides use of a modulator molecule of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder.

In one aspect, the invention provides use of a nucleic acid of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder.

In one aspect, the invention provides use of an expression vector of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder.

In one aspect, the invention provides use of a host cell of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder.

In one aspect, the invention provides use of an article of manufacture of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder.

In one aspect, the invention provides use of a kit of the invention in the preparation of a medicament for the therapeutic and/or prophylactic treatment of a disease, such as a cancer, a tumor, a cell proliferative disorder, an immune (such as autoimmune) disorder and/or an angiogenesis-related disorder

In one aspect, the invention provides a method of inhibiting c-met activated cell proliferation, said method comprising contacting a cell or tissue with an effective amount of a modulator molecule of the invention, whereby cell proliferation associated with c-met activation is inhibited.

In one aspect, the invention provides a method of treating a pathological condition associated with dysregulation of c-met activation in a subject, said method comprising administering to the subject an effective amount of a modulator molecule of the invention, whereby said condition is treated.

In one aspect, the invention provides a method of inhibiting the growth of a cell that expresses c-met or hepatocyte growth factor, or both, said method comprising contacting said cell with a modulator molecule of the invention thereby causing an inhibition of growth of said cell. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

In one aspect, the invention provides a method of therapeutically treating a mammal having a cancerous tumor comprising a cell that expresses c-met or hepatocyte growth factor, or both, said method comprising administering to said mammal an effective amount of an a modulator molecule of the invention, thereby effectively treating said mammal. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

In one aspect, the invention provides a method for treating or preventing a cell proliferative disorder associated with increased expression or activity of HGFA, said method comprising administering to a subject in need of such treatment an effective amount of an a modulator molecule of the invention, thereby effectively treating or preventing said cell proliferative disorder. In one embodiment, said proliferative disorder is cancer.

In one aspect, the invention provides a method for treating or preventing a cell proliferative disorder associated with increased expression or activity of c-met or hepatocyte growth factor, or both, said method comprising administering to a subject in need of such treatment an effective amount of a modulator molecule of the invention, thereby effectively treating or preventing said cell proliferative disorder. In one embodiment, said proliferative disorder is cancer.

In one aspect, the invention provides a method for inhibiting the growth of a cell, wherein growth of said cell is at least in part dependent upon a growth potentiating effect of HGFA, said method comprising contacting said cell with an effective amount of a modulator molecule of the invention, thereby inhibiting the growth of said cell. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

In one aspect, the invention provides a method for inhibiting the growth of a cell, wherein growth of said cell is at least in part dependent upon a growth potentiating effect of c-met or hepatocyte growth factor, or both, said method comprising contacting said cell with an effective amount of a modulator molecule of the invention, thereby inhibiting the growth of said cell. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

In one aspect, the invention provides a method of therapeutically treating a tumor in a mammal, wherein the growth of said tumor is at least in part dependent upon a growth potentiating effect of HGFA, said method comprising contacting said cell with an effective amount of a modulator molecule of the invention, thereby effectively treating said tumor. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

In one aspect, the invention provides a method of therapeutically treating a tumor in a mammal, wherein the growth of said tumor is at least in part dependent upon a growth potentiating effect of c-met or hepatocyte growth factor, or both, said method comprising contacting said cell with an effective amount of a modulator molecule of the invention, thereby effectively treating said tumor. In one embodiment, the cell is contacted by HGF expressed by a different cell (e.g., through a paracrine effect).

Methods of the invention can be used to affect any suitable pathological state, for example, cells and/or tissues associated with dysregulation of the HGF/c-met signaling pathway, e.g. through increased HGF activity associated with HGFA activation of HGF. In one embodiment, a cell that is targeted in a method of the invention is a cancer cell. For example, a cancer cell can be one selected from the group consisting of a breast cancer cell, a colorectal cancer cell, a lung cancer cell, a papillary carcinoma cell (e.g., of the thyroid gland), a colon cancer cell, a pancreatic cancer cell, an ovarian cancer cell, a cervical cancer cell, a central nervous system cancer cell, an osteogenic sarcoma cell, a renal carcinoma cell, a hepatocellular carcinoma cell, a bladder cancer cell, a prostate cancer cell, a gastric carcinoma cell, a head and neck squamous carcinoma cell, a melanoma cell and a leukemia cell. In one embodiment, a cell that is targeted in a method of the invention is a hyperproliferative and/or hyperplastic cell. In one embodiment, a cell that is targeted in a method of the invention is a dysplastic cell. In yet another embodiment, a cell that is targeted in a method of the invention is a metastatic cell.

Methods of the invention can further comprise additional treatment steps. For example, in one embodiment, a method further comprises a step wherein a targeted cell and/or tissue (e.g., a cancer cell) is exposed to radiation treatment or a chemotherapeutic agent.

As described herein, HGF/c-met activation is an important biological process the dysregulation of which leads to numerous pathological conditions. Accordingly, in one embodiment of methods of the invention, a cell that is targeted (e.g., a cancer cell) is one in which activation of HGF/c-met is enhanced as compared to a normal cell of the same tissue origin. In one embodiment, a method of the invention causes the death of a targeted cell. For example, contact with a modulator molecule of the invention may result in a cell's inability to signal through the c-met pathway, which results in cell death.

Dysregulation of c-met activation (and thus signaling) can result from a number of cellular changes, including, for example, overexpression of HGF (c-met's cognate ligand) and/or HGFA, and/or increased activation of HGF by HGFA. Accordingly, in some embodiments, a method of the invention comprises targeting a tissue wherein one or more of HGFA, c-met and hepatoctye growth factor, is more abundantly expressed and/or present (e.g., a cancer) as compared to a normal tissue of the same origin. An HGF or c-met-expressing cell can be regulated by HGFA from a variety of sources, i.e. in an autocrine or paracrine manner. For example, in one embodiment of methods of the invention, a targeted cell is contacted/bound by hepatocyte growth factor activated by HGFA expressed in a different cell (e.g., via a paracrine effect). Said different cell can be of the same or of a different tissue origin. In one embodiment, a targeted cell is contacted/bound by HGF activated by HGFA expressed by the targeted cell itself (e.g., via an autocrine effect/loop).

In one aspect, the invention provides compositions comprising one or more modulator molecules of the invention and a carrier. In one embodiment, the carrier is pharmaceutically acceptable.

In one aspect, the invention provides nucleic acids encoding a modulator molecule of the invention. In one embodiment, a nucleic acid of the invention encodes a modulator molecule which is or comprises an antibody or fragment thereof.

In one aspect, the invention provides vectors comprising a nucleic acid of the invention.

In one aspect, the invention provides host cells comprising a nucleic acid or a vector of the invention. A vector can be of any type, for example a recombinant vector such as an expression vector. Any of a variety of host cells can be used. In one embodiment, a host cell is a prokaryotic cell, for example, E. coli. In one embodiment, a host cell is a eukaryotic cell, for example a mammalian cell such as Chinese Hamster Ovary (CHO) cell.

In one aspect, the invention provides methods for making a modulator molecule of the invention. For example, the invention provides a method of making a modulator molecule which is or comprises an antibody (or fragment thereof), said method comprising expressing in a suitable host cell a recombinant vector of the invention encoding said antibody (or fragment thereof), and recovering said antibody.

In one aspect, the invention provides an article of manufacture comprising a container; and a composition contained within the container, wherein the composition comprises one or more modulator molecules of the invention. In one embodiment, the composition comprises a nucleic acid of the invention. In one embodiment, a composition comprising a modulator molecule further comprises a carrier, which in some embodiments is pharmaceutically acceptable. In one embodiment, an article of manufacture of the invention further comprises instructions for administering the composition (for e.g., the modulator molecule) to a subject.

In one aspect, the invention provides a kit comprising a first container comprising a composition comprising one or more modulator molecules of the invention; and a second container comprising a buffer. In one embodiment, the buffer is pharmaceutically acceptable. In one embodiment, a composition comprising a modulator molecule further comprises a carrier, which in some embodiments is pharmaceutically acceptable. In one embodiment, a kit further comprises instructions for administering the composition (for e.g., the modulator molecule) to a subject.

In one aspect the invention provides a method of diagnosing a disease comprising determining the level of HGFA in a test sample of tissue cells by contacting the sample with an antibody of the invention, whereby HGFA bound by the antibody indicates presence and/or amount of HGFA in the sample. In another aspect, the invention provides a method of determining whether an individual is at risk for a disease comprising determining the level of HGFA in a test sample of tissue cell by contacting the test sample with an antibody of the invention and thereby determining the amount of HGFA present in the sample, wherein a higher level of HGFA in the test sample, as compared to a control sample comprising normal tissue of the same cell origin as the test sample, is an indication that the individual is at risk for the disease. In one embodiment of methods of the invention, the level of HGFA is determined based on amount of HGFA polypeptide indicated by amount of HGFA bound by the antibody in the test sample. An antibody employed in the method may optionally be detectably labeled, attached to a solid support, or the like.

In one aspect, the invention provides a method of binding an antibody of the invention to HGFA present in a bodily fluid, for example blood.

In yet another aspect, the invention is directed to a method of binding an antibody of the invention to a cell that expresses and/or is responsive to HGFA, wherein the method comprises contacting said cell with said antibody under conditions which are suitable for binding of the antibody to HGFA and allowing binding therebetween. In one embodiment, binding of said antibody to HGFA on the cell inhibits an HGFA biological function. In one embodiment, said antibody does not inhibit interaction of HGFA with its substrate molecule. In one embodiment, said antibody binds to an HGFA molecule on the cell and inhibits binding of another molecule (such as pro-HGF) to the HGFA molecule.

In one aspect, the invention provides a method of targeting a therapeutic agent to an HGFA-associated tissue in a host, the method comprising administering to the host said therapeutic agent in a form that is linked to an antibody of the invention, whereby the agent is targeted to the HGFA-associated tissue in the host. In one embodiment, the antibody that binds HGFA is capable of specifically binding to HGFA located on a cell (either in vitro or in vivo), for example where HGFA is present on the surface of a cell.
 

Claim 1 of 11 Claims

1. An isolated antibody that specifically binds human hepatocyte growth factor activator, wherein the antibody comprises (i) a CDR-H1 sequence comprising the sequence of SEQ ID NO:21; (ii) a CDR-H2 sequence comprising the sequence of SEQ ID NO:22; (iii) a CDR-H3 sequence comprising the sequence of SEQ ID NO:23; and (iv) the CDR L1, L2 and L3 sequences in SEQ ID NO:54.

 

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