Methods and compositions for treating cancer

Abstract

The present invention provides compositions and methods for treating cancer in a subject by administering to the subject an antibody or fragment thereof that binds oxidized LDL. In some embodiments, the compositions and methods can reduce the size of the tumor, reduce macrophage infiltration, and / or inhibit metastasis.

Description

Technical Field

[0001] This invention generally relates to compositions and methods for treating cancer by inhibiting the biological activity of oxidized low-density lipoprotein. Background of the Invention

[0003] Obesity is believed to increase the risk of developing diabetes and several types of cancer. Obesity is associated with a variety of harmful physiological changes, including, for example, adipose tissue dysfunction, low-grade and chronic inflammation, and alterations in lipid metabolism and circulating hormone levels. Oxidized low-density lipoprotein (oxLDL) is a pro-inflammatory mediator formed due to the oxidative modification of LDL in the arterial wall. OxLDL is most commonly associated with the development of atherosclerosis, which can have detrimental cardiovascular (CV) outcomes, including myocardial infarction, stroke, and death. OxLDL signals through various cellular scavenger receptors, including scavenger receptor type A (SR-A), differentiation cluster 36 (CD36), CD68, mucin and lectin-like oxLDL receptor-1 (LOX-1).

[0004] LOX-1 is expressed on endothelial cells, but is also found in macrophages, smooth muscle cells, fibroblasts, and platelets. Furthermore, LOX-1 is secreted in a soluble form. Several studies have shown that oxLDL signaling via LOX-1 plays a major role in the development and progression of atherosclerosis. This invention provides methods and compositions for treating cancer using an oxLDL-dependent mechanism. Summary of the Invention

[0005] In one aspect, the present invention provides a method for inhibiting macrophage infiltration into a tumor in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody or a fragment thereof that binds to oxidized low-density lipoprotein (oxLDL) (i.e., an anti-oxLDL antibody or a fragment thereof).

[0006] In another aspect, the present invention provides a method for inhibiting tumor metastasis in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody or fragment thereof that binds to oxidized low-density lipoprotein (oxLDL) (i.e., an anti-oxLDL antibody or fragment thereof).

[0007] In some implementations, the subject is diagnosed with a tumor.

[0008] In some embodiments, the tumor is a LOX-1 positive tumor. In other embodiments, the tumor is positive for one or more of LOX-1, SR-A, CD36, CD38, and mucin.

[0009] In some embodiments, the tumor is selected from the group consisting of: ovarian cancer, bladder urothelial carcinoma, renal clear cell carcinoma, rectal adenocarcinoma, colonic adenocarcinoma, prostate adenocarcinoma, breast epithelial cell tumor, glioblastoma, pancreatic cancer, and esophageal cancer.

[0010] In some embodiments, the method reduces the growth rate of the tumor, macrophage infiltration of the tumor, and / or the metastatic potential of the tumor.

[0011] In another aspect, the present invention provides a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody or a fragment thereof that binds to oxidized low-density lipoprotein (oxLDL).

[0012] In another aspect, the present invention provides a method for treating cancer in a subject, the method comprising administering to the subject:

[0013] (a) The initial anticancer therapy selected from a group consisting of chemotherapy, radiotherapy, and immunotherapy, and the therapeutically effective dose.

[0014] (b) Therapeutic effective amounts of antibodies or fragments thereof that bind to oxidized low-density lipoprotein (oxLDL).

[0015] In some embodiments, the cancer is LOX-1 positive. In other embodiments, the cancer is positive for one or more of LOX-1, SR-A, CD36, CD38, and mucin.

[0016] In some implementations, the cancer is selected from the group consisting of: ovarian cancer, bladder urothelial carcinoma, renal clear cell carcinoma, rectal adenocarcinoma, colonic adenocarcinoma, prostate adenocarcinoma, breast epithelial cell tumor, glioblastoma, pancreatic cancer, and esophageal cancer.

[0017] In some implementations, the cancer is a blood cancer, including, for example, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute monocytic leukemia, non-Hodgkin's lymphoma, and multiple myeloma.

[0018] In some implementations, the antibody or a fragment thereof is administered as an adjuvant to the initial anticancer therapy.

[0019] In some implementations, the subject is diagnosed with hyperlipidemia, type 2 diabetes, or metabolic syndrome.

[0020] In some embodiments of any of the foregoing aspects, the anti-oxLDL antibody or a fragment thereof inhibits the binding of oxLDL to LOX-1. In other embodiments, the antibody or a fragment thereof inhibits the binding of oxLDL to one or more of SR-A, CD36, CD38, and mucin.

[0021] In some embodiments of any of the foregoing aspects, the antibody or a fragment thereof binds to oxLDL with an affinity at least 10, 50, 100, or 1,000 times greater than that for native (unoxidized) LDL.

[0022] In some implementations, the antibody is a human antibody, a humanized antibody, a mouse antibody or a rabbit antibody, or fragments thereof.

[0023] In some embodiments, the antibody or fragment thereof includes at least one light chain complementarity-determining region (LCDR) substantially identical to an LCDR selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:10.

[0024] In some embodiments, the antibody or fragment thereof includes at least one heavy chain complementarity-determining region (HCDR) substantially identical to the LCDR selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.

[0025] In some embodiments, the antibody or fragment thereof contains a variable heavy region (V) that is substantially the same as that in SEQ ID NO:11. H ), and the variable light region (V) that is substantially the same as SEQ ID NO:12 L (or both)

[0026] In some embodiments, the antibody or fragment thereof comprises with

[0027] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVSSISVGGHRTYYADSVKGRSTISRDNSKNTLYLQMNSLRAEDTAVYYCARIRVGPSGGAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:3) substantially identical heavy chain, and

[0028] QSVLTQPPSASGTPGQRVTISCSGSNTNIGKNYVSWYQQLPGTAPKLLIYANSNRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDASLNGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:4) substantially identical light chain, or both.

[0029] The heavy chain may be composed of

[0030]

[0031] (SEQ ID NO:2) encodes essentially the same polynucleotide sequence.

[0032] In some embodiments, the antibody is orticumab. In other embodiments, the antibody fragment is a fragment of orticumab.

[0033] In some embodiments, the antibody is administered intravenously at an initial dose of at least 5 mg / kg, followed by multiple subsequent doses of at least 2 mg / kg / week, at least 2.5 mg / kg / two weeks, or at least 6 mg / kg / month. In other embodiments, the antibody is administered subcutaneously at a dose of approximately 330 mg / month for at least three months.

[0034] As used herein, "adjuvant therapy" refers to a second therapy administered to treat a primary disease (i.e., the clinical indication for which the initial therapy is targeted), a secondary disease requiring treatment to increase / maximize the effectiveness of the initial therapy, to reduce side effects of treatment or the primary disease, and / or to prevent disease recurrence. In some embodiments, the antibody or antibody fragment is administered as an adjuvant to treat a primary disease (e.g., cancer or tumor) or to reduce plasma concentrations of oxLDL.

[0035] As used herein, “administering” and / or “administer” refers to any route of delivery of a pharmaceutical composition to a patient. Routes of delivery may include noninvasive oral (oral), topical (dermal), transmucosal (nasal, buccal / sublingual, vaginal, ocular, and rectal) and inhalation routes, as well as parenteral routes and other methods known in the art. Parenteral routes refer to delivery routes typically associated with injection, including intraorbital, infusion, intra-arterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intravertebral, intrasternal, intrasheath, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal routes. Through parenteral routes, the composition may be in the form of a solution or suspension for infusion or injection, or as a lyophilized powder.

[0036] As used herein, the term "effective amount" refers to the amount of a pharmaceutical composition used to relieve at least one or more symptoms of a disease or condition, and involves an adequate amount of the pharmaceutical composition to provide the desired effect. As used herein, the phrase "therapeutic effective amount" means an adequate amount of the composition that provides a reasonable benefit / risk ratio for treating the condition, applicable to any medical treatment.

[0037] A significant therapeutic or preventative reduction in symptoms is defined as a measurement of, for example, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 125%, at least about 150%, or more, compared to the condition of a control or untreated subject or the subject prior to administration of the peptides described herein. Measurable or detectable parameters include clinically detectable markers of the disease, such as elevated or decreased levels of biomarkers, and parameters associated with clinically accepted scales for the symptoms or markers of atherosclerosis. However, it should be understood that the total daily dosage of the compositions and formulations disclosed herein will be determined by the attending physician within the bounds of reasonable medical judgment. The exact amount required will vary depending on factors such as the type of disease being treated, the subject's sex, age, and weight.

[0038] "Subject," "individual," "animal," "patient," or "mammal" refers to any subject requiring diagnosis, prognosis, or treatment, particularly a mammalian subject. Mammal subjects include, but are not limited to, humans, domestic animals, farm animals, zoo animals, sporting animals, pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, and dairy cows; primates such as apes, monkeys, orangutans, and chimpanzees; canines such as dogs and wolves; felines such as cats, lions, and tigers; equines such as horses, donkeys, and zebras; food animals such as cattle, pigs, and sheep; ungulates such as deer and giraffes; rodents such as mice, rats, hamsters, and guinea pigs; etc. In some implementations, the mammal is a human subject.

[0039] "Substantially identical" means a nucleic acid or amino acid sequence that shares at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with a second nucleic acid or amino acid sequence when optimally aligned, for example, using the methods described below. "Substantially identical" can be used to refer to sequences of various types and lengths, such as full-length sequences, epitopes or immunogenic peptides, functional domains, coding and / or regulatory sequences, exons, introns, promoters, and genomic sequences. The percentage of identity between two polypeptide or nucleic acid sequences is determined in various ways within the scope of the art, for example, using publicly available computer software such as: Smith-Waterman alignment (Smith, TF and MS-Waterman (1981) J Mol Biol 147:195-7); “BestFit” (Smith and Waterman, Advances in Applied Mathematics, 482-489 (1981)), as incorporated in GeneMatcher Plus. TMSchwarz and Dayhof (1979) Atlas of Protein Sequence and Structure, Dayhof, MO, Ed, pp. 353-358; BLAST program (basic local alignment search tool); (Altschul, SF, W. Gish, et al. (1990) J Mol Biol215:403-10), BLAST-2, BLAST-P, BLAST-N, BLAST-X, WU-BLAST-2, ALIGN, ALIGN-2, CLUSTAL, or Megalign (DNASTAR) software. Furthermore, those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms required to achieve maximum alignment in the length of the compared sequences. Generally, for proteins, the length of the compared sequences will be at least 10 amino acids, preferably 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, 300, or 350 amino acids. Or 400 amino acids or more. For nucleic acids, the length of the compared sequences will typically be at least 25, 50, 100, 125, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 800, 900, 1000, 1100, or 1200 or more. It should be understood that when comparing DNA and RNA sequences, thymine nucleotides are equivalent to uracil nucleotides to determine sequence identity. Conserved substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.

[0040] Attached Figure Description

[0041] The invention can be better understood by referring to the following accompanying drawings. The components in the drawings are not necessarily drawn to scale; the focus is on illustrating the principles of this disclosure. In the drawings, the same reference numerals indicate corresponding parts in all different views.

[0042] Figure 1 This is a line graph comparing the binding of otesulamab to natural (unoxidized) LDL and MDA-LDL.

[0043] Figure 2A This is a bar graph showing the release of MCP-1 from cells treated with otesulamab and control antibodies. Figure 2B It is a bar graph showing the cumulative release of MCP-1 over time.

[0044] Figure 3These are micrographs of protein blots stained with IκBα.

[0045] Figures 4A-4B This illustrates macrophage infiltration in a mouse model of atherosclerosis after treatment with otetumumab or a control antibody. Figure 4A ) and total plaque load ( Figure 4B (The image is missing.) Detailed Implementation

[0046] It is well known that a high-fat diet and lipid-related diseases increase the incidence of certain types of cancer, including breast cancer, prostate cancer, colon cancer, and liver cancer. Plasma lipids have long been associated with several types of cancer. In particular, significantly higher levels of total cholesterol (TC), triglycerides (TG), and LDL have been found in some breast cancer patients.

[0047] A high-fat diet can induce a shift to a favorable microenvironment that supports cancer cell development and growth. This shift is caused by changes in the hormonal environment or cell membrane properties (which are due to alterations in lipid composition) or by modulating the immune response to tumor cells. Furthermore, as with other lipid-related metabolic diseases (e.g., obesity and NASH), oxidative stress leads to the oxidation of LDL to oxLDL, resulting in significant mutagenic and carcinogenic effects. Various clinical studies have highlighted the role of oxLDL in carcinogenesis, and a positive correlation between elevated serum oxLDL concentrations and cancer risk has been reported in pancreatic, colon, breast, and esophageal cancers. In addition, in clinical studies of colorectal cancer (CRC) patients, lipid oxidation products (including oxLDL) induced by oxidative stress have been investigated as potential biomarkers for CRC development. Although this study did not observe a significant difference in serum oxLDL levels between CRC patients and healthy controls, oxLDL levels were significantly higher in patients with early-stage primary tumors compared to those with advanced primary tumor progression (Diakowska et al., Gastroenterol. Res. Pract. 2015:146819(2015)).

[0048] OxLDL-mediated signaling via LOX-1 leads to the upregulation of adhesion proteins, other pro-inflammatory mediators, and pro-angiogenic factors, all of which are pathogenic in cancer. Among these pathogenic signaling molecules are monocyte chemokine-1 (MCP-1) and nuclear factor-κB (NFκB). MCP-1 is a chemokine that drives macrophage recruitment to inflammatory areas, and its expression is correlated with the degree of tumor-associated macrophage (TAM) infiltration. NFκB is a transcription factor that acts as a major regulator of pro-inflammatory gene expression. NFκB activity is upregulated in inflammatory diseases, including cancer, where it contributes to the transformation of normal cells into tumor cells, tumor cell survival, and the persistent inflammatory cycle in cancer. Importantly, oxLDL-Lox-1-mediated signaling has been associated with both MCP-1 upregulation and NFκB activity.

[0049] This invention is partly based on the characterization of the anticancer properties of anti-oxLDL antibodies. Direct interference with oxLDL can inhibit LOX-1-mediated tumorigenesis and / or progression. This invention is also based on the finding that otesulamab specifically binds to the oxidized form of LDL (i.e., oxLDL) relative to natural / unoxidized LDL. Ootesulamab is a fully human recombinant monoclonal IgG1 antibody with an affinity of approximately 8 ± 6 nM for oxLDL.

[0050] Anti-oxLDL antibody

[0051] As discussed herein, the present invention incorporates an anti-oxLDL antibody. A particularly useful anti-oxLDL antibody is otetumab. The synthesis and characterization of otetumab are described in WO2009 / 08205 and referred to therein as antibody 2D03. WO2009 / 08205 is hereby incorporated herein by reference in its entirety.

[0052] Figure 2 of WO 2009 / 08205 illustrates the amino acid sequences of the 2D03 heavy chain and the 2D03 light chain, with the complementarity-determining region (CDR) underlined.

[0053] WO 2007 / 025781 (which is hereby incorporated herein by reference in its entirety) describes the invention as also including an antibody that selectively binds to an oxidized LDL epitope selectively bound by antibody 2D03 (i.e., otesulamb), and also including an antibody comprising at least one, two, three, four, five or all six amino acid sequences having the corresponding CDR of antibody 2D03. (See, for example, pages 16, 29, and 30.) Furthermore, antibodies having three or four CDRs having sequences corresponding to the 2D03 antibody CDRs preferably have all three heavy chain CDRs or all three light chain CDRs having sequences of the corresponding CDRs of antibody 2D03; thus, this aspect of the invention includes antibodies comprising three light chain CDRs having sequences of the corresponding three light chain CDRs of antibody 2D03 or three heavy chain CDRs having sequences of the corresponding three heavy chain CDRs of antibody 2D03; more preferably, the antibody comprises three light chain CDRs and three heavy chain CDRs having sequences of the corresponding CDRs of antibody 2D03; if the antibody does not contain all of the sequences of the corresponding CDRs of antibody 2D03... The six CDRs, preferably some or all of 1, 2, 3, 4 or 5 “different” CDRs, contain variants of the sequence of the corresponding CDR of antibody 2D03 (with regard to “variant”, WO2007 / 025781 means that the variant has at least 50% sequence identity with the corresponding CDR sequence, more preferably at least 70%, even more preferably at least 80%, at least 90%, or at least 95%; most preferably, the variant has 96%, 97%, 98%, or 99% sequence identity with the corresponding CDR sequence of antibody 2D03; generally, the “variant” CDR sequence differs from the corresponding CDR sequence of antibody 2D03 by 5, 4, 3, 2 or only 1 amino acid residue); and this aspect of the invention includes antibody 2D03.

[0054] Heavy chain complementarity-determining regions (HCDRs) 1 (HCDR1), 2 (HCDR2), and 3 (HCDR3) are listed in SEQ ID Nos: 5, 6, and 7, respectively; and light chain complementarity-determining regions (LCDRs) 1 (LCDR1), 2 (LCDR2), and 3 (LCDR3) are listed in SEQ ID Nos: 8, 9, and 10, respectively. Otetuzumab contains the variable heavy region (VH) amino acid sequence of SEQ ID No: 11 and the variable light region (VL) amino acid sequence of SEQ ID No: 12. Otetuzumab contains the heavy chain amino acid sequence of SEQ ID No: 3 and the light chain amino acid sequence of SEQ ID No: 4.

[0055] HCDR1 is FSNAWMSWVRQAPG (SEQ ID NO:5).

[0056] HCDR2 is SSISVGGHRTYYADSVKGR (SEQ ID NO:6).

[0057] HCDR3 is ARIRVGPSGGAFDY (SEQ ID NO:7).

[0058] LCDR1 is CSGSNTNIGKNYVS (SEQ ID NO:8).

[0059] LCDR2 is ANSNRPS (SEQ ID NO:9).

[0060] LCDR3 is CASWDASLNGWV (SEQ ID NO:10).

[0061] Variable repetition zone (V H )yes:

[0062] EVQLLESGGG LVQPGGSLRL SCAASGFTFS NAWMSWVR QA PGKGLEWVSS ISVGGHRTYYADSVKGRSTI SRDNSKNTLY LQMNSLRAED TAVYYCARIR VGPSGGAFDY WGQGTLVTVS (SEQ ID NO: 11).

[0063] Variable light region (V L )yes:

[0064] QSVLTQPPSA SGTPGQRVTI SCSGSNTNIG KNYVSWYQQL PGTAPKLLIY ANSNRPSGVPDRFSGSKSGT SASLAISGLR SED EADYYCA SWDASLNGWV FGGGTKLTVL (SEQ ID NO: 12).

[0065] In some embodiments, the antibody or antibody fragment comprises, or is substantially composed of, the sequences listed in SEQ ID NO:1 and / or 2.

[0066] In some embodiments, the present invention provides an antibody or antibody fragment that binds to oxLDL and one or more of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 as listed in SEQ ID No.:5-10.

[0067] In some embodiments, the present invention provides an antibody comprising at least one CDR having the amino acid sequence of the corresponding CDR of otelizumab. More preferably, the antibody has two, three, four, or five CDRs having the sequence of the corresponding CDR of otelizumab. If the antibody has three or four CDRs having the sequence of the corresponding CDR of otelizumab, then preferably the antibody has all three heavy chain CDRs or all three light chain CDRs having the sequence of the corresponding CDR of otelizumab. Thus, this aspect of the method includes an antibody comprising three light chain CDRs having the sequence of the corresponding three light chain CDRs of otelizumab or three heavy chain CDRs having the sequence of the corresponding three heavy chain CDRs of otelizumab. Even more preferably, the antibody comprises three light chain CDRs and three heavy chain CDRs having the sequence of the corresponding CDR of otelizumab.

[0068] If the antibody does not contain all six CDRs having the sequence of the corresponding CDR of otelizumab, then variants containing some or all of one, two, three, four, or five “different” CDRs are preferred. “Variant” means that the variant has at least 50%, more preferably at least 70%, even more preferably at least 80%, at least 90%, or at least 95% sequence identity with the corresponding CDR. Most preferably, the variant has 96%, 97%, 98%, or 99% sequence identity with the corresponding CDR of otelizumab. Typically, the “variant” CDR sequence differs from the corresponding CDR of otelizumab by five, four, three, two, or only one amino acid residue.

[0069] Specifically, the present invention provides antibodies containing one or more of "HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3", covering embodiments in which the antibody contains one, any two, any three, any four, any five, or all six of the CDRs (i.e., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3). For example, one aspect of the embodiments provides an antibody containing HCDR1 as listed in SEQ ID NO:5. Another aspect provides an antibody containing HCDR2 as listed in SEQ ID NO:6. Another aspect provides an antibody containing HCDR3 as listed in SEQ ID NO:7. Another aspect provides an antibody containing LCDR1 as listed in SEQ ID NO:8. Another aspect provides an antibody containing LCDR2 as listed in SEQ ID NO:9. Another aspect provides an antibody containing LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains HCDR1 as listed in SEQ ID NO:5 and HCDR2 as listed in SEQ ID NO:6. Another aspect provides that the antibody contains HCDR1 as listed in SEQ ID NO:5 and HCDR3 as listed in SEQ ID NO:7. Another aspect provides that the antibody contains HCDR1 as listed in SEQ ID NO:5 and LCDR1 as listed in SEQ ID NO:8. Another aspect provides that the antibody contains HCDR1 as listed in SEQ ID NO:5 and LCDR2 as listed in SEQ ID NO:9. Another aspect provides that the antibody contains HCDR1 as listed in SEQ ID NO:5 and LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains HCDR2 as listed in SEQ ID NO:6 and HCDR3 as listed in SEQ ID NO:7. Another aspect provides that the antibody contains HCDR2 as listed in SEQ ID NO:6 and LCDR1 as listed in SEQ ID NO:8. Another aspect provides that the antibody contains HCDR2 as listed in SEQ ID NO:6 and LCDR2 as listed in SEQ ID NO:9. Another aspect provides that the antibody contains HCDR2 as listed in SEQ ID NO:6 and LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains HCDR3 as listed in SEQ ID NO:7 and LCDR1 as listed in SEQ ID NO:8.Another aspect provides that the antibody contains HCDR3 as listed in SEQ ID NO:7 and LCDR2 as listed in SEQ ID NO:9. Another aspect provides that the antibody contains HCDR3 as listed in SEQ ID NO:7 and LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains LCDR1 as listed in SEQ ID NO:8 and LCDR2 as listed in SEQ ID NO:9. Another aspect provides that the antibody contains LCDR1 as listed in SEQ ID NO:8 and LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains LCDR2 as listed in SEQ ID NO:9 and LCDR3 as listed in SEQ ID NO:10. Another aspect provides that the antibody contains HCDR1, HCDR2, and HCDR3 as listed in SEQ ID NO:5-7 respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, and LCDR1 as listed in SEQ ID NO:5, 6, and 8 respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, and LCDR2 as listed in SEQ ID NO.:5, 6, and 9, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, and LCDR3 as listed in SEQ ID NO.:5, 6, and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, and LCDR1 as listed in SEQ ID NO.:5, 7, and 8, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, and LCDR2 as listed in SEQ ID NO.:5, 7, and 9, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, and LCDR3 as listed in SEQ ID NO.:5, 7, and 10, respectively. Another aspect provides that the antibody contains HCDR1, LCDR1, and LCDR2 as listed in SEQ ID NO.:5, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR1, LCDR1, and LCDR3 as listed in SEQ ID NO.:5, 8, and 10, respectively. Another aspect provides that the antibody contains HCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, and LCDR1 as listed in SEQ ID NO.:6, 7, and 8, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, and LCDR2 as listed in SEQ ID NO.:6, 7, and 9, respectively.Another aspect provides that the antibody contains HCDR2, HCDR3, and LCDR3 as listed in SEQ ID NO.:6, 7, and 10, respectively. Another aspect provides that the antibody contains HCDR2, LCDR1, and LCDR2 as listed in SEQ ID NO.:6, 8, and 10, respectively. Another aspect provides that the antibody contains HCDR2, LCDR1, and LCDR3 as listed in SEQ ID NO.:6, 8, and 10, respectively. Another aspect provides that the antibody contains HCDR2, LCDR2, and LCDR3 as listed in SEQ ID NO.:6, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR3, LCDR1, and LCDR2 as listed in SEQ ID NO.:7, 8, and 9, respectively. Another aspect provides that the antibody contains HCDR3, LCDR1, and LCDR3 as listed in SEQ ID NO.:7, 8, and 10, respectively. Another aspect provides that the antibody contains HCDR3, LCDR2, and LCDR3 as listed in SEQ ID NO.:7, 9, and 10, respectively. Another aspect provides that the antibody contains LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:8-10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, and LCDR1 as listed in SEQ ID NO.:5-8, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, and LCDR2 as listed in SEQ ID NO.:5-7 and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, LCDR1, and LCDR2 as listed in SEQ ID NO.:5-7 and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, LCDR1, and LCDR2 as listed in SEQ ID NO.:5, 6, 8, and 9, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, LCDR1, and LCDR3 as listed in SEQ ID NO.:5, 6, 8, and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 6, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, LCDR1, and LCDR2 as listed in SEQ ID NO.:5, 7, 8, and 9, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, LCDR1, and LCDR3 as listed in SEQ ID NO.:5, 7, 8, and 10, respectively.Another aspect provides that the antibody contains HCDR1, HCDR3, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 7, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR1, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 8, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, LCDR1, and LCDR2 as listed in SEQ ID NO.:6-9, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, LCDR1, and LCDR3 as listed in SEQ ID NO.:6-8 and 10, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, LCDR2, and LCDR3 as listed in SEQ ID NO.:6, 7, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR2, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:6, 8, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR3, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:7-10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, LCDR1, and LCDR2 as listed in SEQ ID NO.:5-9, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, LCDR1, and LCDR3 as listed in SEQ ID NO.:5-8 and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, LCDR2, and LCDR3 as listed in SEQ ID NO.:5-7, 9, and 10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 6, 8-10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR3, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:5, 7-10, respectively. Another aspect provides that the antibody contains HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:6-10, respectively. Another aspect provides that the antibody contains HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 as listed in SEQ ID NO.:5-10, respectively.

[0070] When preparing and using variants of any polypeptide sequence (e.g., CDR) provided herein, it should be understood that a given amino acid may be substituted with residues having similar physiological and chemical characteristics, for example, replacing one aliphatic residue with another (such as replacing one with Ile, Val, Leu, or Ala), or replacing one polar residue with another (such as between Lys and Arg; Glu and Asp; or Gln and Asn). Other such conserved substitutions, such as substitution of an entire region having similar hydrophobic characteristics or substitution of residues having similar side chain volumes, are well known. Isolated antibodies containing conserved amino acid substitutions may be tested in any of the assays described herein to confirm the desired activity, as determined by assays described elsewhere herein.

[0071] Amino acids can be grouped according to the similarity of the properties of their side chains (in Allehninger, in Biochemistry, 2nd edition, pp. 73-75, Worth Publishers, New York (1975)): (1) Nonpolar: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) Nonpolar: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) Acidic: Asp (D), Glu (E); (4) Basic: Lys (K), Arg (R), His (H). Alternatively, naturally occurring residues can be grouped based on common side-chain properties into several groups: (1) hydrophobic: ortholeucine, Met, Ala, Val, Leu, Ile, Phe, Trp; (2) neutral-hydrophilic: Cys, Ser, Thr, Asn, Gln, Ala, Tyr, His, Pro, Gly; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues affecting chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr, Phe, Pro, His, or hydroxyproline. Non-conservative substitution would require replacing members of one of these categories with members of another.

[0072] Particularly preferred conservative substitutions for the variants described herein are as follows: Ala substituted with Gly or Ser; Arg substituted with Lys; Asn substituted with Gln or His; Asp substituted with Glu or Asn; Cys substituted with Ser; Gln substituted with Asn; Glu substituted with Asp; Gly substituted with Ala or Pro; His substituted with Asn or Gln; Ile substituted with Leu or Val; Leu substituted with Ile or Val; Lys substituted with Arg, Gln, or Glu; Met substituted with Leu, Tyr, or Ile; Phe substituted with Met, Leu, or Tyr; Ser substituted with Thr; Thr substituted with Ser; Trp substituted with Tyr or Phe; Tyr substituted with Phe or Trp; and / or Phe substituted with Val, Tyr, Ile, or Leu. Generally, conservative substitutions encompass residue exchanges with residues having similar physicochemical properties (i.e., replacing another hydrophobic amino acid with a hydrophobic residue).

[0073] Any cysteine ​​residues that do not participate in maintaining the proper conformation of the isolated peptide as described herein may be substituted (usually with serine) to improve the oxidative stability of the molecule and prevent undesirable cross-linking. Conversely, cysteine ​​bonds may be added to the isolated peptide as described herein to improve its stability or promote polymerization.

[0074] In some embodiments, the antibody as described herein may comprise naturally occurring amino acids commonly found in polypeptides and / or proteins produced by living organisms, such as Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M), Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q), Asp (D), Glu (E), Lys (K), Arg (R), and His (H). In some embodiments, the antibody may comprise substituted amino acids. Non-limiting examples of alternative amino acids include D-amino acids, β-amino acids, homocysteine, phosphoserine, phosphothreonine, phosphotyrosine, hydroxyproline, γ-carboxyglutamic acid, hippuric acid, octahydroindole-2-carboxylic acid, pepsinine, 1,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid, penicillamine (3-mercapto-D-valine), ornithine, citrulline, α-methyl-alanine, p-benzoylphenylalanine, p-aminophenylalanine, p-fluorophenylalanine, phenylglycine, propargylglycine, sarcosine, and tert-butylglycine, diaminobutyric acid, 7-hydroxy-tetrahydroisoquinoline carboxylic acid, naphthylalanine, and biphenylalanine. Amino acids, cyclohexylalanine, amino-isobutyric acid, valine, leucine, tert-leucine, tetrahydroisoquinoline carboxylic acid, pipercoic acid, phenylglycine, homophenylalanine, cyclohexylglycine, dehydroleucine, 2,2-diethylglycine, 1-amino-1-cyclopentane carboxylic acid, 1-amino-1-cyclohexane carboxylic acid, amino-benzoic acid, amino-naphthoic acid, γ-aminobutyric acid, difluorophenylalanine, piperidinecarboxylic acid, α-aminobutyric acid, thienyl-alanine, tert-butylglycine, trifluorovaline, hexafluoroleucine, fluorinated analogs, azide-modified amino acids, alkyne-modified amino acids, cyano-modified amino acids, and their derivatives.

[0075] In some embodiments, the antibody may be modified, for example, by adding a portion to one or more amino acids. In some embodiments, the antibody may comprise one or more moiety molecules, such as one or more moiety molecules per peptide, two or more moiety molecules per peptide, five or more moiety molecules per peptide, ten or more moiety molecules per antibody, or more moiety molecules per antibody. In some embodiments, the antibody as described herein may comprise one or more types of modifications and / or moieties, such as one type of modification, two types of modifications, three types of modifications, or more types of modifications. Non-limiting examples of modifications and / or moieties include PEGylation; glycosylation; HES-ization; ELP-ization; esterification; acetylation; amidation; capping modification; cyanoation; phosphorylation; and cyclization. In some embodiments, capping modification may include N-terminal acetylation, N-terminal acylation, and N-terminal formylation. In some embodiments, capping modification may include C-terminal amidation, introduction of C-terminal alcohol, aldehyde, ester, and thioester moieties.

[0076] Treatment

[0077] This invention also provides a method for treating or preventing cancer by administering to a subject a therapeutically effective amount of an antibody or antibody fragment that specifically binds to oxLDL. This invention also provides a method for preventing metastasis in a subject diagnosed with cancer by administering to a subject a therapeutically effective amount of an antibody or antibody fragment that specifically binds to oxLDL. In some embodiments, the binding of the antibody or antibody fragment inhibits or blocks at least one biological function of oxLDL. In some embodiments, the cancer is pancreatic cancer, breast cancer, colorectal cancer including rectal adenocarcinoma and colonic adenocarcinoma, ovarian cancer, bladder urothelial carcinoma, clear cell renal cell carcinoma, prostate cancer (prostate adenocarcinoma), and in some embodiments, the cancer cells express and / or are identified as expressing LOX-1. In some embodiments, the antibody or antibody fragment inhibits, reduces, or blocks the binding of oxLDL to LOX-1. In some embodiments, the antibody or antibody fragment inhibits, reduces, or blocks the binding of oxLDL to SR-A, CD36, CD38, and / or mucin. In some embodiments, the antibody or antibody fragment is oteltuzumab, a fragment of oteltuzumab, a derivative of oteltuzumab, or any anti-oxLDL antibody or antibody fragment described herein.

[0078] In various embodiments, the composition to be administered in the disclosed methods is formulated for delivery via any route of administration. For example, the methods include administration via aerosol, nasal, oral, mucosal, percutaneous, parenteral, or enteral routes. "Parenteral" refers to routes of administration generally associated with injection, including intraoral, infusion, intra-arterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrasheath, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal injection. via the parenteral route, the composition may be in the form of a solution or suspension for infusion or injection, or as a lyophilized powder. via the parenteral route, the composition may be in the form of a solution or suspension for infusion or injection. via the enteral route, the pharmaceutical composition may be in the form of tablets, gel capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres, or lipid vesicles or polymer vesicles that allow controlled release. Typically, the composition is administered by injection.

[0079] Typically, the methods disclosed herein produce an effective amount of antioxLDL in subjects to produce a plasma concentration of at least 4 μg / mL, most preferably at least 12 μg / mL.

[0080] The method of the present invention may include subcutaneous administration of the disclosed antibody or antibody fragment at about 330 mg / month to the subject for about 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months or longer, and the subject is an adult. Other embodiments provide administration of the antibody or antibody fragment at not less than 2 mg / kg / week (166 mg for a patient with an average weight of 83 kg); most preferably, about 4 mg / kg / week (332 mg for a patient with an average weight of 83 kg) weekly. In another aspect, the anti-oxLDL antibody composition is administered every two weeks at >2.5 mg / kg / two weeks (e.g., 208 mg for a patient with an average weight of 83 kg). In yet another aspect, the anti-oxLDL antibody composition is administered monthly at about 6 mg / kg / month (e.g., about 498 mg for a patient with an average weight of 83 kg). For example, monthly dosing may be performed for 12 months or 3 months. Other embodiments provide administration of the antibody or antibody fragment to the subject at an initial dose of 800-900 mg, 900-1000 mg, 1000-1100 mg, 1100-1200 mg, 1200-1300 mg, 1300-1400 mg, 1400-1500 mg, or 1500-1600 mg. In some aspects, the effective amount described herein comprises an initial dose of approximately 1000-1500 mg of otesulamab, followed by subsequent doses of the antibody administered weekly at 700-900 mg for 2, 3, 4, or 5 weeks and / or even monthly for 1, 2, or 3 months.

[0081] Other embodiments provide administration of an anti-oxLDL antibody or fragment in progressively increasing doses. In this embodiment, an exemplary (starting) dose of a single-dose administration of an antibody against oxLDL (e.g., otelizumab) is between 0.005 and 0.01 mg / kg (e.g., intravenously); and other exemplary dose levels to be administered in a single-dose administration are between 0.01 and 0.15 mg / kg, between 0.15 and 0.75 mg / kg, between 0.75 and 2.5 mg / kg, between 2.5 and 7.5 mg / kg, and between 7.5 and 30 mg / kg (e.g., intravenously). For example, the starting dose of a single intravenous administration of otelizumab is 0.007 mg / kg; and other exemplary doses in subsequent single intravenous administrations may be 0.05, 0.25, 1.25, 5.0, or 15.0 mg / kg. In another embodiment, a single subcutaneous dose of the antibody is administered between 0.5 and 5 mg / kg, and multiple subcutaneous doses are also administered between 0.5 and 5 mg / kg. For example, a subcutaneous dose of 1.25 mg / kg of antibody is administered. In various embodiments, the dose is administered within a specific hourly range per day in each administration, and each dose in multiple-dose treatments (e.g., 4 doses, 3 doses, 5 doses, or 6 doses) is administered weekly at a time window of ±1 day. In another example, an antibody (e.g., otelasumab) is administered to a human subject at a concentration between 300 mg and 450 mg (e.g., 360 mg), optionally followed by another dose between 300 mg and 450 mg (e.g., 360 mg), wherein the second dose is administered at least 70 days (up to 91 days) after the first dose. The antibody can be formulated at a concentration of 100-170 mg / mL (e.g., 150 mg / mL) for subcutaneous administration without further dilution or dilution to a large volume for intravenous infusion.

[0082] Other implementation schemes include administering the drug to subjects at doses of approximately 10-50 μg / cycle, 50-100 μg / cycle, 100-150 μg / cycle, 150-200 μg / cycle, 100-200 μg / cycle, 200-300 μg / cycle, 300-400 μg / cycle, 400-500 μg / cycle, 500-600 μg / cycle, 600-700 μg / cycle, 700-800 μg / cycle, 800-900 μg / cycle, 900-1000 μg / cycle, 1000-1100 μg / cycle, 1100-1200 μg / cycle, 1200-1300 μg / cycle, 1300-1400 μg / cycle, 140 μg / cycle, etc. Effective antibody doses within the ranges of 0-1500 μg / cycle, 1500-1600 μg / cycle, 1600-1700 μg / cycle, 1700-1800 μg / cycle, 1800-1900 μg / cycle, 1900-2000 μg / cycle, 2000-2100 μg / cycle, 2100-2200 μg / cycle, 2200-2300 μg / cycle, 2300-2400 μg / cycle, 2400-2500 μg / cycle, 2500-2600 μg / cycle, 2600-2700 μg / cycle, 2700-2800 μg / cycle, 2800-2900 μg / cycle, or 2900-3000 μg / cycle. Cycles can be one day, one week, one month, or other time lengths. One aspect is that the antibody (e.g., otesulamab) is administered at a frequency of weekly, bi-weekly, or monthly per cycle, for any of the aforementioned doses.

[0083] In some embodiments, the method includes administering an anti-oxLDL antibody (e.g., otelasumab) to a subject for 1-5 days, 1-5 weeks, 1-5 months, or 1-5 years. For example, the antibody may be administered to the subject in 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 doses, with each dose spaced at least 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, or combinations thereof. In other embodiments, a second dose may be administered approximately 2-3 weeks or approximately 3 weeks after the first dose, and a third dose may be administered approximately 5-6 weeks or approximately 6 weeks after the first dose. In yet another embodiment, a second dose may be administered approximately 2-3 months, approximately 2 months, approximately 3 months, or approximately 4 months after the first dose, and a third dose may be administered approximately 4-6 months, approximately 5-6 months, approximately 5 months, or approximately 6 months after the first dose.

[0084] Pharmaceutical compositions and drugs

[0085] In various embodiments, the present invention provides pharmaceutical compositions for use in said methods. The pharmaceutical composition comprises an anti-oxLDL antibody or a fragment thereof and a pharmaceutically acceptable carrier. As used herein, a “pharmaceutically acceptable carrier” refers to a pharmaceutically acceptable material, composition, or medium that participates in carrying or transporting a target compound from one tissue, organ, or part of the body to another. For example, a carrier can be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof. Examples of excipients include, but are not limited to, starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, wetting agents, emulsifiers, colorants, stripping agents, coating agents, sweeteners, flavoring agents, aroma agents, preservatives, antioxidants, plasticizers, gelling agents, thickeners, hardening agents, setting agents, suspending agents, surfactants, humectants, carriers, stabilizers, and combinations thereof. Generally, the components of the carrier must be “pharmaceutically acceptable” because it must be compatible with the other components of the formulation. It must also be suitable for contact with any tissue or organ it may come into contact with, which means it must not carry the risk of toxicity, irritation, allergic reactions, immunogenicity, or any other complications that outweigh its therapeutic benefits.

[0086] According to the present invention, a pharmaceutical composition can be delivered in a therapeutically effective amount. The precise therapeutically effective amount is the amount of composition that will produce the most effective result in terms of therapeutic efficacy in a given subject. This amount will vary depending on a variety of factors, including but not limited to the characteristics of the therapeutic compound (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, disease type and stage, general physical condition, responsiveness to a given dose, and type of drug), the nature of one or more pharmaceutically acceptable carriers in the formulation, and the route of administration. Those skilled in the art of clinical and pharmacology will be able to determine the therapeutically effective amount through routine experiments, such as by monitoring the subject's response to the administered compound and adjusting the dose accordingly. For further guidance, see Remington: The Science and Practice of Pharmacy (Gennaro, ed., 20th edition, Williams & Wilkins PA, USA) (2000).

[0087] Antibody preparation

[0088] This invention is based in part on the use of anti-oxLDL antibodies, fragments, and binding proteins. Modern recombinant library technology is used to prepare therapeutic antibodies against oxLDL. Although mouse hybridoma cells produce large quantities of the same antibodies, these non-human antibodies are recognized as foreign by the human body, and therefore, in addition to triggering allergic reactions, their efficacy and plasma half-life are reduced. To address this problem, one approach is to prepare chimeric antibodies, in which the mouse variable domain of the antibody is transferred to the human constant region, thereby producing antibodies that are predominantly human. A further improvement to this approach is the development of humanized antibodies, in which the mouse antibody region in contact with the antigen, namely the complementarity-determining region (CDR), is transferred to the human antibody framework, thereby producing humanized antibodies. Another approach is to use recombinant technology to produce fully human antibodies, a technology that does not rely on animal immunization to produce specific antibodies. Instead, recombinant libraries contain a large number of pre-prepared antibody variants, and the library is likely to contain at least one antibody specific to any antigen. Phage display systems can be used, in which antibody fragments are expressed and displayed as fusions with phage coat proteins on the surface of filamentous phage particles, while the phage display system simultaneously carries genetic information encoding the displayed molecule. Phage fragments exhibiting antibody specificity for a particular antigen can be selected by binding to the antigen in question. The isolated phage can then be amplified, and optionally, the gene encoding the variable domain of the selected antibody can be transferred to other antibody forms, such as full-length immunoglobulins, and expressed in large quantities using appropriate vectors and host cells well known in the art. The antibody specificity displayed on the phage particle may vary. The most common forms are Fab and single-chain (scFv), both containing the variable antigen-binding domain of the antibody. The single-chain form consists of a variable heavy domain (VH) connected to a variable light domain (VL) via a flexible linker. The displayed antibody specificity is converted to a soluble form, such as Fab or scFv, before being used as an analytical reagent or therapeutic agent, and thus analyzed. In subsequent steps, antibody fragments identified as having the desired characteristics can be transferred to other forms, such as full-length antibodies.

[0089] Antibody production from hybridoma

[0090] Cell fusion is performed using standard procedures well known to those skilled in the art of immunology. Fusion partner cell lines and methods used for fusion and selection of hybridomas, as well as for screening mAbs, are well known in the art. See, for example, Ausubel, Harlow, and Colligan, the contents of which are incorporated herein by reference in their entirety.

[0091] Anti-oxLDL antibodies can be produced in large quantities by injecting antibody-secreting hybridoma or transfected tumor cells into the peritoneal cavity of mice, harvesting ascites containing high titers of mAb after an appropriate time, and isolating the mAb from the ascites. For this in vivo production using mAbs from non-mouse hybridomas (e.g., rats or humans), the hybridoma cells are preferably grown in irradiated or thymus-agnostic nude mice. Alternatively, antibodies can be produced by culturing hybridoma or transfected tumor cells in vitro and isolating the secreted mAb from the cell culture medium or by recombinant synthesis in eukaryotic or prokaryotic cells.

[0092] Recombinant expression of antibodies

[0093] Recombinant mouse or chimeric mouse-human or human-human antibodies that inhibit oxidized LDL can be provided using known techniques based on the teachings provided herein, according to the present invention. See, for example, Ausubel et al., eds., Current Protocols in Molecular Biology, Wiley Interscience, NY (1987, 1992, 1993); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989).

[0094] The DNA encoding anti-oxLDL antibodies can be genomic DNA or cDNA encoding at least one of the heavy chain constant region (Hc), heavy chain variable region (Hc), light chain variable region (Lv), and light chain constant region (Lc). A convenient alternative to using chromosomal gene fragments as the DNA source encoding the mouse V region antigen-binding segment is to construct chimeric immunoglobulin genes using cDNA, for example, as reported by Liu et al. (Proc. Natl. Acad. Sci., USA 84:3439 (1987) and J. Immunology 139:3521 (1987)). Using cDNA requires combining gene expression elements suitable for the host cell with the gene to achieve the desired protein synthesis. Using cDNA sequences is advantageous over genomic sequences (which contain introns) because cDNA sequences can be expressed in bacteria or other hosts lacking a suitable RNA splicing system.

[0095] Example

[0096] The following embodiments are not intended to limit the scope of the claims to the invention, but are intended to be examples of certain implementations. Any modifications of the exemplified methods that may occur to those skilled in the art are intended to fall within the scope of the invention.

[0097] Example 1: Otetrazol specifically binds to oxLDL

[0098] The binding specificity of osteosulinab to various forms of LDL was investigated. In this experiment, the binding of osteosulinab to native (unoxidized) LDL and malondialdehyde-modified LDL (MDA-LDL) was measured across different concentrations, and the dissociation constant (K0) was calculated. d MDA-LDL was chosen for this experiment because it is an endogenous LDL species and is believed to reflect the naturally occurring levels of oxLDL.

[0099] The binding of otesulamb to malondialdehyde (MDA)-modified and native human LDL prepared from donor serum was tested in an ELISA assay. LDL samples were immobilized on plates and antibody concentrations were titrated. 50 μl of MDA-LDL or native LDL was coated onto each well of a multi-well plate and diluted to 2 μg / mL with PBS + 1 mM EDTA. The plates were incubated overnight at 4°C. The purified antibody was diluted, titrated in ELISA blocking buffer (0.2% skim milk powder), and then applied. The plates were incubated at room temperature for 1 hour. Binding antibodies were detected using horseradish peroxidase (HRP)-conjugated rabbit anti-human IgG antibody (P0214 DAKO).

[0100] Figure 1 Exemplary binding curves of otesulamab (0.01–50 μg / ml) to native LDL and MDA-LDL are provided. Ootesulamab lacks any significant affinity for native LDL but exhibits robust and specific binding to MDA-LDL. d It is approximately 8±6 nM.

[0101] Example 2: Otetrazol blocks oxLDL-induced release of MCP-1 from macrophages

[0102] The effect of otesulamab on oxLDL-induced macrophage release of MCP-1 was measured to assess its potential anti-inflammatory effects. MCP-1 is a macrophage chemical inducer secreted by activated macrophages in inflammatory areas and used to enhance the inflammatory response. MCP-1 expression is associated with tumor-associated macrophage infiltration.

[0103] Freshly isolated CD14+ macrophages were pre-activated with 0.1 ng / mL lipopolysaccharide (LPS) to generate pro-inflammatory M1 macrophages. After 20 hours of pre-activation, otesulamb, FITC-8, or an equal volume of medium was added to a final concentration of 40 nM, and the cells were cultured for another 24 hours. Culture supernatant was collected, and MCP-1 levels were analyzed using a cell counting bead array (CBA; BD Bioscience, Franklin Lakes, NJ, USA). Figure 2AThe mean (+ / - SD) or pooled and normalized data are shown from two different donors in two different experiments with treatment in triplicate. Mean MCP-1 values ​​for samples not treated with the antibody were normalized. Statistical analysis was performed by ANOVA, followed by Tukey-Kramer multiple comparison tests using GraphPadInstat 3 software (***p<0.001). (Stored data; Report BI 209-68) The results indicate that otesulamb treatment significantly reduced MCP-1 secretion in ox-LDL-stimulated macrophages compared to treatment with the control antibody (FITC-8) and the mediator control (“Ab-free”).

[0104] In another experiment, macrophages derived from human monocytes were grown for 14 days in the presence of human serum containing oxLDL. These cells were then treated with either control IgG (FITC-8) or otelasuma antibody for specified time periods. The supernatant was collected daily and MCP-1 levels were analyzed. Figure 2B This indicates that in the presence of otesulamb, MCP-1 levels did not increase over time, as observed in cultures treated with the control antibody, thus demonstrating that otesulamb effectively blocked MCP-1 secretion. For example, after 4 days (96 hours), otesulamb blocked the increase of MCP-1, resulting in a reduction of up to 60% in macrophage MCP-1 release compared to cells treated with control IgG.

[0105] Example 3: Otetazidine inhibits NFκB signaling by increasing IκBα expression.

[0106] NFκB is a well-known transcriptional regulator of genes that respond to stress stimuli in most cell types and typically upregulate immune responses and are involved in inflammation. In unstimulated cells, NFκB is primarily located in the cytoplasm and is chelated by inhibitors, including IκB, which mask its nuclear localization signals. Upon cellular stimulation, IκB is rapidly phosphorylated by IκB kinase (“IKK”) and subsequently ubiquitinated and degraded. Once freed from IκB inhibition, NFκB rapidly translocates to the nucleus.

[0107] The effect of otetumab on oxLDL signaling in the NFκB pathway was investigated. Macrophages were stimulated with lipopolysaccharide (LPS) in the presence of oxLDL and treated with otetumab or a mutant otetumab-like antibody lacking significant affinity for oxLDL and / or native LDL. Monocytes were isolated from healthy subjects and incubated in the absence or presence of 0.3 ng / mL LPS. Cells were simultaneously treated with a control antibody, otetumab, or a mutant otetumab. Cells were then harvested and immunoblotted to obtain the total IκBα or control protein (actin). Whole-cell lysates from primary human monocytes were extracted with RIPA buffer (150 mM NaCl, 1.0% IGEPA CA-630, 0.5% v / v sodium deoxycholate, 0.1% w / v SDS, and 50 mM Tris-chloride, pH 8.0). After centrifugation to remove cell debris, aliquots of 20,000 g of supernatant were subjected to 10% SDS / PAGE, followed by protein transfer to a Hybond-C additional nitrocellulose filter (Amersham Biosciences, Piscataway, NJ). The filter was incubated with the primary antibody at room temperature. The bound antibody was visualized by chemiluminescence (SuperSignal Substrate; Thermo Fisher Scientific, Waltham, MA) using a 1:5000 dilution of horseradish peroxidase-conjugated donkey anti-rabbit IgG or donkey anti-mouse IgG (Jackson Immuno Research, West Grove, PA). The filter was exposed to Kodak X-Omat BlueXB-1 film at room temperature for 1–60 seconds.

[0108] like Figure 3 As shown, otetumumab treatment revealed IκBα expression in LPS-stimulated macrophages and control macrophages. These results indicate that otetumumab possesses anti-inflammatory activity in oxLDL-stimulated macrophages by inhibiting NFκB through increased IκBα expression.

[0109] Example 4: Otetazidine inhibits macrophage infiltration

[0110] Based on the aforementioned biochemical evidence, the ultimate effect of otesulamab on oxLDL-induced macrophage infiltration was investigated. A well-characterized atherosclerotic mouse model was used. Apobec-1- / - / LDLR- / - mice with a C57BL / 6 background from Jackson Laboratories (BarHarbor, Maine) (these mice expressed full-length apoB-100 in their LDL particles and had 3-fold higher plasma apoB-100 levels compared to LDLR- / - mice) were fed a randomly provided high-cholesterol diet (15% cholesterol, 21% fat; Lactamin AB, Kimstad Sweden) from 4 weeks of age. One week before the first treatment (24 weeks of age), the diet was changed to a normal diet. One week later (25 weeks of age), a group of mice was sacrificed as a baseline control (baseline 25 weeks). The remaining animals were either untreated (control, 29 weeks) or administered 1 mg (0.5 mL) of control IgG antibody (fluorescein isothiocyanate-8 (FITC-8) or otesulamab via intraperitoneal (IP) injection. The injections were repeated weekly for a total of 3 doses, and the mice were sacrificed 2 weeks (29 weeks old) after the last injection.

[0111] The efficacy of otesulam in limiting macrophage infiltration was evaluated using two endpoints. Figure 4A This indicates that 4 weeks of otesulamb treatment after plaque formation significantly reduced the observed number of infiltrating macrophages compared to untreated or control subjects treated with control IgG. This inhibition led to beneficial physiological changes in treated subjects. Macrophage infiltration in the brachiocephalic artery was assessed by MOMA2 monoclonal antibody staining (*p<0.05 compared to FITC-8). Figure 4B This indicates that otesulamab treatment significantly reduced plaque burden compared to pretreatment conditions and various controls. Plaque burden in the descending aorta was assessed by Oil Red O staining, and the percentage of total plaque area per unit area of ​​the descending aorta was calculated. P-values ​​were calculated relative to fluorescein isothiocyanate-8 (FITC-8) (***p<0.001 compared to FITC-8).

[0112] All references cited herein are incorporated herein by reference in their entirety as if fully explained. Unless otherwise defined, the technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention pertains. Allen et al., Remington: The Science and Practice of Pharmacy, 22nd ed., Pharmaceutical Press (September 15, 2012); Hornyak et al., Introduction to Nanoscience and Nanotechnology, CRC Press (2008); Singleton and Sainsbury, Dictionary of Microbiology and Molecular Biology, 3rd ed., revised edition, J. Wiley & Sons (New York, NY 2006); Smith, March's Advanced Organic Chemistry Reactions, Mechanisms and Structure, 7th ed., J. Wiley & Sons (New York, NY 2013); Singleton, Dictionary of DNA and Genome Technology, 3rd ed., Wiley-Blackwell (November 28, 2012); and Green and Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor Laboratory Press). (Harbor, NY 2012) provides general guidance to those skilled in the art regarding many of the terms used in this application. For references on how to prepare antibodies, see Greenfield, Antibodies: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press (Cold Spring Harbor NY, 2013); And Milstein, Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion, Eur. J. Immunol. July 1976 6(7):511-9; Queen and Selick, Humanized immunoglobulins, U.S. Patent No. 5,585,089 (December 1996); and Riechmann et al., Reshaping human antibodies for therapy, Nature March 24, 1988, 332(6162):21-7.

[0113] The foregoing description sets forth the invention and its methods of use in several embodiments. Those skilled in the art will be able to make changes and modifications to the content described herein without departing from its spirit and scope. While the invention may have various embodiments in different forms, preferred embodiments are shown in the accompanying drawings and will be described in detail herein. It should be understood that this disclosure should be considered as an illustration of the principles of the invention and is not intended to limit the broad aspects of the invention to the illustrated embodiments. Unless otherwise stated, all features, elements, components, functions, and steps described with respect to any embodiment provided herein are intended to be freely combined and substituted with those features, elements, components, functions, and steps from any other embodiment. Therefore, it should be understood that the description is for illustrative purposes only and should not be considered as a limitation on the scope of the invention.

[0114] The invention has been described broadly and generally herein. Each species and subgenus falling within the general scope of the disclosure is also part of the method. This includes a general description of the method with accompanying conditions or negative limitations, to remove any subject from the genus regardless of whether the removed material is specifically described herein.

[0115] Other embodiments are defined in the following claims. Furthermore, as the features or aspects of the method are described according to the Markush group, those skilled in the art will recognize that the invention has therefore also been described according to any individual member or subgroup of the Markush group. <110> Abcentra, LLC <120> Methods and compositions for treating cancer <130> 51473-006WO2 <150> US 62 / 934,078 <151> 2019-11-12 <160> 12 <170> PatentIn version 3.5 <210> 1 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> Polynucleotide sequence encoding the heavy chain of an oxLDL-specific antibody <400> 1 gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcagt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcaagt attagtgttg gtggacatag gacatattat 180 gcagattccg tgaagggccg gtccaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatga acagcctgag agccgaggac actgccgtgt attactgtgc acggatacgg 300 gtgggtccgt ccggcggggc ctttgactac tggggccagg gtacactggt caccgtgagc 360 tcagcctcca ccaagggccc atcggtcttc cccctggcac cctcctccaa gagcacctct 420 gggggcacag cggccctggg ctgcctggtc aaggactact tccccgaacc ggtgacggtg 480 tcgtggaact caggcgccct gaccagcggc gtgcacacct tcccggctgt cctacagtcc 540 tcaggactct actccctcag cagcgtggtg accgtgccct ccagcagctt gggcacccag 600 acctacatct gcaacgtgaa tcacaagccc agcaacacca aggtggacaa gaaagttgag 660 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 720 ggaccgtcag tcttctctctt ccccccaaaaa cccaaggaca ccctcatgat ctcccggacc 780 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 840 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 900 aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 960 aaggagtca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaccatc 1020 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggat 1080 gagctgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 1140 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 1200 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1260 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1320 acgcagaaga gcctctccct gtctccgggt aaa 1353 <210> 2 <211> 648 <212> DNA <213> Artificial Sequence <220> <223> Polynucleotide sequence encoding the light chain of an oxLDL-specific antibody <400> 2 cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60 tcctgctctg gaagcaacac caacattggg aagaactatg tatcttggta tcagcagctc 120 ccaggaacgg cccccaaact cctcatctat gctaatagca atcggccctc aggggtccct 180 gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccgg 240 tccgaggatg aggctgatta ttactgtgcg tcatgggatg ccagcctgaa tggttgggta 300 ttcggcggag gaaccaagct gacggtccta ggtcagccca aggctgcccc ctcggtcact 360 ctgttcccgc cctcctctga ggagcttcaa gccaacaagg ccacactggt gtgtctcata 420 agtgacttct acccgggagc cgtgacagtg gcctggaagg cagatagcag ccccgtcaag 480 gcgggagtgg agaccaccac accctccaaa caaagcaaca acaagtacgc ggccagcagc 540 tatctgagcc tgacgcctga gcagtggaag tcccacagaa gctacagctg ccaggtcacg 600 catgaaggga gcaccgtgga gaagacagtg gcccctacag aatgttca 648 <210> 3 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> Heavy chain <400> 3 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Val Gly Gly His Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Ser Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Arg Val Gly Pro Ser Gly Gly Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 4 <211> 216 <212> PRT <213> Artificial Sequence <220> <223> Light chain <400> 4 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Asn Thr Asn Ile Gly Lys Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ala Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp Ala Ser Leu 85 90 95 Asn Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 5 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> HCDR1 <400> 5 Phe Ser Asn Ala Trp Met Ser Trp Val Arg Gln Ala Pro Gly 1 5 10 <210> 6 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> HCDR2 <400> 6 Ser Ser Ile Ser Val Gly Gly His Arg Thr Tyr Tyr Tyr Ala Asp Ser Val 1 5 10 15 Lys Gly Arg <210> 7 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> HCDR3 <400> 7 Ala Arg Ile Arg Val Gly Pro Ser Gly Gly Ala Phe Asp Tyr 1 5 10 <210> 8 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> LCDR1 <400> 8 Cys Ser Gly Ser Asn Thr Asn Ile Gly Lys Asn Tyr Val Ser 1 5 10 <210> 9 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> LCDR2 <400> 9 Ala Asn Ser Asn Arg Pro Ser 1 5 <210> 10 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> LCDR3 <400> 10 Cys Ala Ser Trp Asp Ala Ser Leu Asn Gly Trp Val 1 5 10 <210> 11 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> Variable heavy region <400> 11 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Val Gly Gly His Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Ser Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Arg Val Gly Pro Ser Gly Gly Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser 115 120 <210> 12 <211> 110 <212> PRT <213> Artificial Sequence <220> <223> Variable light region <400> 12 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Asn Thr Asn Ile Gly Lys Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ala Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp Ala Ser Leu 85 90 95 Asn Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110

Claims

1. Use of an antibody or fragment thereof in the manufacture of a medicament for inhibiting macrophage infiltration into a tumor in a subject in need thereof, wherein the antibody or fragment thereof binds to oxidized low density lipoprotein (oxLDL) and comprises a heavy chain complementarity determining region (HCDR)1 consisting of the sequence of SEQ ID NO: 5, a HCDR2 consisting of the sequence of SEQ ID NO: 6, and a HCDR3 consisting of the sequence of SEQ ID NO: 7, a light chain complementarity determining region (LCDR)1 consisting of the sequence of SEQ ID NO: 8, a LCDR2 consisting of the sequence of SEQ ID NO: 9, and a LCDR3 consisting of the sequence of SEQ ID NO:

10.

2. Use of an antibody or fragment thereof in the manufacture of a medicament for inhibiting tumor metastasis in a subject in need thereof, wherein the antibody or fragment thereof binds to oxidized low density lipoprotein (oxLDL) and comprises a heavy chain complementarity determining region (HCDR)1 consisting of the sequence of SEQ ID NO: 5, a HCDR2 consisting of the sequence of SEQ ID NO: 6, and a HCDR3 consisting of the sequence of SEQ ID NO: 7, a light chain complementarity determining region (LCDR)1 consisting of the sequence of SEQ ID NO: 8, a LCDR2 consisting of the sequence of SEQ ID NO: 9, and a LCDR3 consisting of the sequence of SEQ ID NO:

10.

3. The use of claim 1 or 2, wherein the tumor is LOX-1 positive.

4. The use of claim 1 or 2, wherein the tumor is a CD36 positive tumor.

5. The use of claim 1 or 2, wherein the tumor is selected from the group consisting of: ovarian cancer, bladder urothelial carcinoma, renal clear cell carcinoma, rectal adenocarcinoma, colon adenocarcinoma, prostate adenocarcinoma, breast epithelial cell tumor, glioblastoma, pancreatic cancer, and esophageal cancer.

6. The use of claim 1 or 2, wherein the use reduces the growth rate of the tumor.

7. Use of an antibody or fragment thereof in the manufacture of a medicament for treating cancer in a subject in need thereof, wherein the antibody or fragment thereof binds to oxidized low density lipoprotein (oxLDL) and comprises a heavy chain complementarity determining region (HCDR)1 consisting of the sequence of SEQ ID NO: 5, a HCDR2 consisting of the sequence of SEQ ID NO: 6, and a HCDR3 consisting of the sequence of SEQ ID NO: 7, a light chain complementarity determining region (LCDR)1 consisting of the sequence of SEQ ID NO: 8, a LCDR2 consisting of the sequence of SEQ ID NO: 9, and a LCDR3 consisting of the sequence of SEQ ID NO:

10.

8. The use of claim 7, wherein the antibody or fragment thereof is to be administered with a therapeutically effective amount of an initial anti-cancer therapy selected from the group consisting of chemotherapy, radiation therapy, and immunotherapy.

9. The use of claim 7, wherein the cancer is LOX-1 positive.

10. The use of claim 7, wherein the cancer is a CD36 positive tumor.

11. The use of claim 7, wherein the cancer is selected from the group consisting of: ovarian cancer, bladder urothelial carcinoma, renal clear cell carcinoma, rectal adenocarcinoma, colon adenocarcinoma, prostate adenocarcinoma, breast epithelial cell tumor, glioblastoma, pancreatic cancer, and esophageal cancer.

12. The use of claim 7, wherein the cancer is a hematological cancer.

13. The use of claim 12, wherein the hematological cancer is selected from the group consisting of: acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, acute monocytic leukemia, non-Hodgkin lymphoma, and multiple myeloma.

14. The use of claim 7, wherein the antibody or fragment thereof is an adjuvant to an initial anti-cancer therapy.

15. The use of claim 7, wherein the subject is diagnosed with hyperlipidemia, type 2 diabetes, or metabolic syndrome.

16. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof inhibits binding of oxLDL to LOX-1.

17. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof binds to oxLDL with an affinity that is at least 100 times greater than the affinity for native LDL.

18. The use of any one of claims 1, 2, or 7, wherein the antibody is a human antibody.

19. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof comprises a variable heavy region (V H ) of SEQ ID NO: 11, a variable light region (V L ) of SEQ ID NO: 12, or both.

20. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 3, a light chain of SEQ ID NO: 4, or both.

21. The use of any one of claims 1, 2, or 7, wherein the antibody is bapineuzumab.

22. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof is administered intravenously at an initial dose of at least 5 mg / kg, followed by a plurality of subsequent doses.

23. The use of claim 22, wherein the subsequent doses are each at least 2 mg / kg / week.

24. The use of claim 22, wherein the subsequent doses are each at least 2.5 mg / kg / two weeks.

25. The use of claim 22, wherein the subsequent doses are each at least 6 mg / kg / month.

26. The use of any one of claims 1, 2, or 7, wherein the antibody or fragment thereof is administered subcutaneously at a dose of 330 mg / month for at least three months.

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