Immune-Derived Moieties Reactive Against Bioactive Lipids, and Methods of Making and Using Same
Compositions and methods for producing monoclonal antibodies and their derivatives reactive against bioactive lipid targets are described. These compositions include derivatized lipids, each of which comprises a bioactive lipid that having a polar head group and at least one hydrocarbon chain (e.g., a lysolipid such as lysophosphatidic acid or sphingosine-1-phosphate) in which a carbon atom has been derivatized with a pendant reactive group; immunogens made by linking a derivatized lipid to a carrier moiety (e.g., a carrier protein, polyethylene glycol, colloidal gold, alginate, or a silicone bead); monoclonal antibodies and derivatives produced by immunizing an animal with such an immunogen; and therapeutic and diagnostic compositions containing such antibodies and antibody derivatives. Methods for making such derivatized lipids, immunogens, and monoclonal antibodies and derivatives, methods for detecting such antibodies once generated, and therapeutic and diagnostic methods for using such antibodies and derivatives, are also described.
1 . An isolated immune-derived moiety reactive against a bioactive lipid, wherein the moiety is derived from an antibody produced by immunizing an immune competent mammalian host with an immunogen comprising a derivatized bioactive lipid covalently linked to a carrier, wherein the derivatized bioactive lipid comprises a polar head group and at least one hydrocarbon chain, wherein the derivatized bioactive lipid is selected from the group consisting of:
(a) a derivatized lipid wherein a carbon atom within the hydrocarbon chain of the bioactive lipid is derivatized with an optionally protected pendant sulfhydryl group, wherein the optionally protected pendant sulfhydryl group is optionally appended at a position selected from the group consisting of an SN-1 position and an SN-2 position of the glycerol backbone of the bioactive lipid; and
(b) a derivatized lipid wherein the bioactive lipid is a lysolipid and a carbon atom within the hydrocarbon chain is derivatized with an optionally protected pendant reactive group.
2 . An isolated immune-derived moiety according to claim 1 is selected from the group consisting of an antibody, an antibody fragment, an antibody variant, and an antibody derivative.
3 . An isolated immune-derived moiety according to claim 1 , wherein the bioactive lipid is selected from the group consisting of a lysophosphatidic acid, a lysophosphatidic acid precursor, and a lysophosphatidic acid metabolite.
4 . An isolated immune-derived moiety according to claim 1 that is an antibody selected from the group consisting of a chimeric monoclonal antibody, a human monoclonal antibody, and a humanized monoclonal antibody.
5 . A composition comprising an isolated immune-derived moiety according to claim 1 in a carrier, optionally a pharmaceutically acceptable carrier.
6 . A composition according to claim 1 , wherein the immune-derived moiety is bispecific.
7 . A composition according to claim 6 , wherein the immune-derived moiety is reactive against both a lysophosphatidic acid and a sphingolipid.
8 . A method of decreasing the effective concentration of a bioactive lipid in a subject, comprising administering to the subject an immune-derived moiety according to claim 1 that is reactive against a bioactive lipid in an amount effective to decrease the effective concentration of said bioactive lipid, wherein the bioactive lipid is a non-sphingoid bioactive lipid.
9 . A method according to claim 16 , wherein the subject is a mammal, optionally a human.
10 . A method of treatment, comprising administering to a subject in need of therapeutic or prophylactic treatment an amount of an immune-derived moiety according to claim 1 effective to accomplish such treatment.
11 . A method according to claim 10 , wherein the immune-derived moiety is a monoclonal antibody reactive against a lysolipid, optionally selected from the group consisting of a lysophosphatidic acid, a lysophosphatidic acid precursor, and a lysophosphatidic acid metabolite.
12 . A method according to claim 10 , wherein the subject is a mammal, optionally a human.
13 . A method selected from the group consisting of:
(a) a method of inhibiting proliferation of a cancer cell, comprising administering to a subject having a cancer responsive to lysophosphatidic acid signaling an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to inhibit proliferation of the cancer cell, wherein the cancer cell is optionally selected from the group consisting of a renal carcinoma cell, a pancreatic carcinoma cell, a melanoma cell, a lung carcinoma cell, a neuroblastoma cell, a hepatocellular carcinoma cell, a glioblastoma multiforme cell, a breast cancer cell, an ovarian cancer cell, a prostate cancer cell, a colorectal cancer cell, and a leukemia cell;
(b) a method of inhibiting migration of a cancer cell, comprising administering to a subject having a cancer responsive to lysophosphatidic acid signaling an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to inhibit migration of the cancer cell, wherein the cancer cell is optionally selected from the group consisting of a renal carcinoma cell, a pancreatic carcinoma cell, a melanoma cell, a lung carcinoma cell, a neuroblastoma cell, a hepatocellular carcinoma cell, a glioblastoma multiforme cell, a breast cancer cell, an ovarian cancer cell, a prostate cancer cell, a colorectal cancer cell, and a leukemia cell;
(c) a method of inhibiting tumor metastasis, comprising administering to a subject having a potentially metastatic cancer an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to inhibit metastasis of the tumor, wherein the tumor is optionally selected from the group consisting of renal carcinoma, pancreatic carcinoma, melanoma, lung carcinoma, neuroblastoma, hepatocellular carcinoma, glioblastoma multiforme, breast cancer, ovarian cancer, prostate cancer, colorectal cancer, and leukemia;
(d) a method of inhibiting angiogenesis in a tumor, comprising administering to a subject having a tumor an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to inhibit angiogenesis in the tumor, wherein the tumor is optionally selected from the group consisting of renal carcinoma, pancreatic carcinoma, melanoma, lung carcinoma, neuroblastoma, hepatocellular carcinoma, glioblastoma multiforme, breast cancer, ovarian cancer, prostate cancer, colorectal cancer, and leukemia;
(e) a method of increasing apoptosis of a cancer cell, comprising contacting the cancer cell with an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to increase apoptosis of the cancer cell, wherein the cancer cell is optionally selected from the group consisting of a renal carcinoma cell, a pancreatic carcinoma cell, a melanoma cell, a lung carcinoma cell, a neuroblastoma cell, a hepatocellular carcinoma cell, a glioblastoma multiforme cell, a breast cancer cell, an ovarian cancer cell, a prostate cancer cell, a colorectal cancer cell, and a leukemia cell; and
(f) a method of enhancing the apoptotic effect of a cytotoxic agent against a cancer cell, comprising contacting the cancer cell with an amount of an immune-derived moiety according to claim 3 that is reactive with a lysophosphatidic acid effective to enhance the apoptotic effect of a cytotoxic agent against the cancer cell, wherein the cancer cell is optionally selected from the group consisting of a renal carcinoma cell, a pancreatic carcinoma cell, a melanoma cell, a lung carcinoma cell, a neuroblastoma cell, a hepatocellular carcinoma cell, a glioblastoma multiforme cell, a breast cancer cell, an ovarian cancer cell, a prostate cancer cell, a colorectal cancer cell, and a leukemia cell.
14 . A method of treating cancer, comprising administering to an animal having or suspected of having cancer a therapeutically effective amount of an isolated immune-derived moiety reactive against a lysophosphatidic acid to decrease the effective concentration of lysophosphatidic acid in the animal, thereby effecting treatment, wherein the cancer is optionally selected from the group consisting of renal carcinoma, pancreatic carcinoma, melanoma, lung carcinoma, neuroblastoma, hepatocellular carcinoma, glioblastoma multiforme, breast cancer, ovarian cancer, prostate cancer, colorectal cancer, and leukemia.
15 . A method according to claim 15 further comprising administering a cytotoxic agent.
16 . A method of administration, comprising administering an immune-derived moiety according to claim 1 to a subject in need of treatment with the immune-derived moiety.
17 . A method according to claim 16 , wherein the immune-derived moiety is a monoclonal antibody reactive against a lysolipid, optionally a lysophosphatidic acid, and wherein the monoclonal antibody is optionally selected from the group consisting of a chimeric monoclonal antibody, a human monoclonal antibody, and a humanized monoclonal antibody.
18 . A method according to claim 16 , wherein the immune-derived moiety is selected from the group consisting of an antibody, an antibody fragment, an antibody variant, and an antibody derivative.
19 . A method according to claim 16 , wherein the subject is a mammal, optionally a human, and wherein administration is via a route selected from the group consisting of topical administration (optionally via a route selected from the group consisting of a transdermal route, an epidermal ophthalmic route, an intrauterine route, a vaginal route, a rectal route, a pulmonary route, an intratracheal route, and an intranasal route), oral administration, and parenteral administration (optionally via a route selected from the group consisting of intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, and intracranial administration).