Compositions and methods for targeting and killing alpha-v beta-3-positive cancer stem cells (CSCS) and treating drug resistant and metastatic cancers

Abstract

To provide compositions and methods for treating and killing alpha-v beta-3-positive cancer stem cells (CSCs) and treating drug resistant and metastatic cancers.SOLUTION: Provided are compositions and methods for treating or ameliorating an advanced cancer such as a drug resistant or metastatic cancer which express αvβ3 polypeptides on their cell surfaces, or for killing cancer stem cells (CSCs) which express αvβ3 polypeptides on their cell surfaces, by using human or humanized antibodies capable of specifically binding cell surface-expressed αvβ3 polypeptides, wherein an Fc region has a selective affinity to human FcγRI (CD64), but not to other FcγRs on the effector cells such as macrophages, neutrophils and dendritic cells.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This Patent Cooperation Treaty (PCT) international application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 62 / 882,296, filed August 2, 2019. The foregoing application is expressly incorporated by reference herein in its entirety for all purposes. All publications, patents, and patent applications cited herein are expressly incorporated by reference herein for all purposes.

[0002] Technical Field The present invention generally relates to immunology and oncology. In an alternative embodiment, compositions and methods are provided for treating or ameliorating advanced cancers (e.g., drug-resistant or metastatic cancers) that express αvβ3 polypeptides on their cell surface or killing cancer stem cells (CSCs) that express αvβ3 polypeptides on their cell surface by using human or humanized antibodies whose Fc region has selective affinity for human FcγRI (CD64) on effector cells (e.g., macrophages, neutrophils, and dendritic cells) and is capable of specifically binding to αvβ3 (avb3) polypeptides expressed on their cell surface. By administering these antibodies to individuals in need, these human or humanized antibodies can treat, ameliorate, or delay the development of advanced or drug-resistant cancers, or cancers caused, initiated, or sustained by cells of advanced or drug-resistant cancers, or cancer stem cells (CSCs). In an alternative embodiment, the administered human or humanized antibody induces an antibody-dependent cell-mediated cytotoxicity (ADCC) response against cells of advanced or drug-resistant cancer, or CSCs. [Background technology]

[0003] background Antibodies induce antibody-dependent cell-mediated cytotoxicity (ADCC) against target cells by utilizing effector cells (e.g., macrophages, natural killer cells, dendritic cells, and neutrophils). To utilize these effector cells, the Fc of the antibody must have affinity for Fcγ receptors (FcγRs) on the effector cells. Summary of the Invention [Means for solving the problem]

[0004] Abstract In an alternative embodiment, - Treating or ameliorating cancer, where appropriate, advanced or drug-resistant cancer; or - Killing cancer stem cells (CSCs), wherein the cancer, the advanced cancer, the drug-resistant cancer, or the CSC expresses an αvβ3 polypeptide on their cell surface; The method comprises administering to an individual in need thereof a human or humanized antibody capable of Fc region-specific binding to the human FcγRI (CD64) receptor but no or substantially no binding to other human FcγRs, and capable of specific binding to cell surface-expressed αvβ3 (avb3) polypeptide; wherein optionally the human FcγRI (CD64) receptor is expressed on the surface of human macrophages, neutrophils and / or dendritic cells; Thereby, a method is provided for inducing an antibody-dependent cell-mediated cytotoxicity (ADCC) response or reaction against cells of said advanced or drug-resistant cancer, or CSCs.

[0005] In an alternative embodiment of the method as provided herein: - the human or humanized antibody comprises the monoclonal antibody (mAb) LM609 (MedImmune), or the mAb having ATCC Accession No. HB9537, or the mAb as described in U.S. Pat. No. 5,753,230;

[0006] - The human or humanized antibody is VITAXIN TM (MedImmune) or MEDI-523;

[0007] - the human or humanized antibody is etaracizumab (or etaratuzumab), or MEDI-522, or ABEGRIN TM (MedImmune) included;

[0008] - the method further comprises administering to the individual in need thereof an additional cancer therapeutic agent or treatment, wherein optionally the additional cancer therapeutic agent comprises paclitaxel;

[0009] The human or humanized antibody is administered to the individual in need thereof at a dose of between about 1 and about 8 mg / kg, or between about 0.5 and about 12 mg / kg.

[0010] The human or humanized antibody is administered intravenously (IV), intrathecally, sublingually, rectally, intravaginally, subcutaneously, or intramuscularly (IM), or is injected or placed near, proximal to, or into a cancer or tumor (e.g., a solid tumor), or an advanced or drug-resistant cancer, or a CSC in situ, or is administered by in situ placement or insertion of an implant comprising the human or humanized antibody.

[0011] - the further cancer medication or treatment is abagovomab, adecatumumab, afutuzumab, alemtuzumab, altumomab, amatuximab, anatumomab, arcitumomab, bavituximab, bectumomab, bevacizumab, bivatuzumab vatuzumab, blinatumomab, brentuximab, cantuzumab, catumaxomab, cetuximab, sitatuzumab, cixutumumab, clivatuzumab, conatumumab, daratumumab, drozitumab, durigotumab tumab, dusigitumab, detumomab, dacetuzumab, dalotuzumab, ecromeximab, elotuzumab, ensituximab, ertumaxomab, farletuzumab, ficlatuzumab, figitumumab, flan votumab), futuximab, ganitumab, gemtuzumab, girentuximab, glembatumumab, ibritumomab, igovomab, imagatuzumab, indatuximab, inotuzumab, intetumumab, ipilimumab,Iratumumab, labetuzumab, lexatumumab, lintuzumab, lorvotuzumab, lucatumumab, mapatumumab, matuzumab, milatuzumab, minretumomab, mitumomab, moxetumomab, narutuzumab natumab, naptumomab, necitumumab, nimotuzumab, nofetumomab, ocaratuzumab, ofatumumab, olaratumab, onartuzumab, oportuzumab, oregovomab, panitumumab, parsatu- zumab mab, patritumumab, pemtumomab, pertuzumab, pintumomab, pritumumab, racotumomab, radretumab, rilotumumab, rituximab, lobatumumab, satumomab, sibrotuzumab, siltuximab ximab, simtuzumab, solitomab, tacatuzumab, taplitumomab, tenatumomab, teprotumumab, tigatuzumab, tositumomab, trastuzumab, tucotuzumab, ublituximab, veltuzumab,The antibody comprises or is an antibody selected from the group consisting of vorsetuzumab, votumumab, zalutumumab, and / or any combination thereof.

[0012] - the additional cancer therapeutic agent or cancer treatment comprises a growth factor inhibitor, optionally wherein the growth factor inhibitor is a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM , mitotic toxin, paclitaxel, taxol, abraxane TM , erlotinib, TARCEVA TM , lapatinib, TYKERB TM , cetuximab, ERBITUX TM , a PD-1 inhibitor, a PD-L1 inhibitor and / or an insulin growth factor inhibitor; and / or

[0013] a plurality of said human or humanized antibodies are pre-incubated ex vivo with human macrophages, neutrophils, monocytes and / or dendritic cells prior to administration to said individual in need thereof, wherein optionally said human macrophages, neutrophils, monocytes and / or dendritic cells are activated human macrophages, neutrophils, monocytes and / or dendritic cells, and optionally said dendritic cells or monocytes are activated as set forth in U.S. Pat. No. 10,023,841.

[0014] In an alternative embodiment, - Treating or ameliorating cancer or tumors, or advanced or drug-resistant cancers, or - Killing cancer stem cells (CSCs), 1. Use of a human or humanized antibody capable of Fc region-specific binding to human FcγRI (CD64) receptor but not or substantially not to other human FcγRs, and capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide, for Here, the cancer or tumor, or the advanced cancer, drug-resistant cancer, or CSC expresses αvβ3 polypeptide on their cell surface, and there is provided a use.

[0015] In an alternative embodiment, - Treating or ameliorating cancer or tumors, or advanced or drug-resistant cancers, or - Killing cancer stem cells (CSCs), A human or humanized antibody capable of Fc region-specific binding to human FcγRI (CD64) receptor but not or substantially not to other human FcγRs, and capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide, for use in Here, the cancer or tumor, or the advanced cancer, drug-resistant cancer, or CSC expresses αvβ3 polypeptide on their cell surface, and a human or humanized antibody is provided.

[0016] The details of one or more exemplary embodiments of the invention are set forth in the description that follows. Other features, objects, and advantages of the invention will become apparent from the detailed description and claims.

[0017] All publications, patents, and patent applications cited herein are expressly incorporated by reference for all purposes.

[0018] The drawings presented herein are illustrative of the exemplary embodiments provided herein and are not meant to limit the scope of the invention as encompassed by the claims. [Brief explanation of the drawings]

[0019] [Figure 1A] Figures 1A-B show the nucleotide and amino acid sequences of the variable regions of antibody VITAX IN™: Figure 1A shows the nucleotide and amino acid sequences of the heavy chain variable region (SEQ ID NO: 1 and SEQ ID NO: 2, respectively), and Figure 1B shows the nucleotide and amino acid sequences of the light chain variable region (SEQ ID NO: 3 and SEQ ID NO: 4, respectively). [Figure 1B] Same as above.

[0020] [Figure 2A] Figures 2A-B show the nucleotide and amino acid sequences of the variable region of monoclonal antibody LM609; Figure 2A shows the nucleotide and amino acid sequences of the heavy chain variable region of LM609 (SEQ ID NO:5 and SEQ ID NO:6, respectively), the variable region spanning amino acids Glu1 to Ala117; Figure 2B shows the nucleotide and amino acid sequences of the light chain variable region of LM609 (SEQ ID NO:7 and SEQ ID NO:8, respectively). [Figure 2B] Same as above.

[0021] [Figure 3] FIG. 3 shows a light chain polypeptide (for pairing with the LM609 heavy chain polypeptide variable region amino acid sequence as shown in FIG. 1A) comprising the variable region amino acid sequence having the nucleotide and amino acid sequences set forth in SEQ ID NO:9 and SEQ ID NO:10, respectively, or a functional fragment thereof. DETAILED DESCRIPTION OF THE INVENTION

[0022] Like reference symbols in the various drawings indicate like elements.

[0023] Detailed Description In an alternative embodiment, compositions and methods are provided for treating or ameliorating cancers or tumors, such as advanced cancers (e.g., drug-resistant cancers) that express αvβ3 polypeptides on their cell surface, or for killing cancer stem cells (CSCs) that express αvβ3 polypeptides on their cell surface, by using (administering) human or humanized antibodies capable of specifically binding to αvβ3 (avb3) polypeptides expressed on the cell surface, whose Fc region has selective affinity for human FcγRI (CD64) on effector cells (e.g., macrophages, neutrophils, and dendritic cells), but not or substantially not for other human FcγRs. By administering these antibodies to an individual in need or performing the methods provided herein, these human or humanized antibodies can treat, ameliorate, or delay the development of the cancer or tumor, or the advanced or drug-resistant cancer, or a cancer caused, initiated, or sustained by cells of the advanced or drug-resistant cancer, or cancer stem cells (CSCs). In an alternative embodiment, the administered human or humanized antibody induces an antibody-dependent cell-mediated cytotoxicity (ADCC) response against cells of advanced or drug-resistant cancer, or CSCs.

[0024] Pharmaceutical Compositions and Formulations In alternative embodiments, provided are pharmaceutical compositions and formulations, for example, comprising human or humanized antibodies capable of specifically binding to αvβ3 (avb3) polypeptide expressed on the cell surface, wherein the Fc region has selective affinity for human FcγRI (CD64) on effector cells (e.g., macrophages, neutrophils, and dendritic cells) but no or substantially no affinity for other human FcγRs, and methods for treating or ameliorating advanced cancers (e.g., drug-resistant cancers that express αvβ3 polypeptide on their cell surface) or killing cancer stem cells (CSCs) that express αvβ3 polypeptide on their cell surface. In alternative embodiments, the pharmaceutical compositions and formulations further comprise an additional therapeutic agent or further comprise immune cells (e.g., macrophages, neutrophils, monocytes and / or dendritic cells or activated forms thereof, optionally including macrophages, neutrophils, monocytes and / or dendritic cells previously incubated ex vivo with the above-described human or humanized antibodies capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide).

[0025] In alternative embodiments, the compositions provided herein and compositions used to practice the methods provided herein are formulated with a pharmaceutically acceptable carrier. In alternative embodiments, the pharmaceutical compositions used to practice the methods provided herein can be administered parenterally, topically, orally, intrathecally, sublingually, rectally, intravaginally, subcutaneously, or intramuscularly (IM), or by any form of local administration (e.g., by aerosol) or transdermally. The pharmaceutical compositions can be formulated in any manner and can be administered in various unit dosage forms, depending on the condition or disease and extent of the illness, the overall medical condition of each patient, the preferred method of administration obtained, and the like. Details regarding techniques for formulation and administration are fully described in the scientific and patent literature. See, for example, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0026] For example, therapeutic agents as provided herein, including anti-αvβ3 (avb3) antibodies, and antibodies used to practice the methods provided herein, can be administered alone or as a component of a pharmaceutical formulation (composition), or simultaneously with, prior to, and / or following administration of another active agent, e.g., a growth factor inhibitor, optionally including a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM , mitotic toxin, paclitaxel, taxol, abraxane TM , erlotinib, TARCEVA TM , lapatinib, TYKERB TM , cetuximab, ERBITUX TM , a PD-1 inhibitor, a PD-L1 inhibitor, or an insulin growth factor inhibitor.

[0027] For example, pharmaceutical compositions and formulations including anti-αvβ3 (avb3) antibodies may be formulated for administration in any convenient way for use in human or veterinary medicine.

[0028] Wetting agents, emulsifiers and lubricants (e.g., sodium lauryl sulfate and magnesium stearate), as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives, buffers and / or antioxidants can also be present in the composition.

[0029] Formulations of the compositions as provided herein used to practice the methods provided herein include those suitable for oral, nasal, topical, parenteral, rectal, subcutaneous, sublingual, intraocular, intramuscular, intrathecal and / or intravaginal administration.

[0030] The above preparations can be conveniently presented in unit dosage form and can be prepared by any method known in the pharmaceutical field.The amount of active ingredient that can be combined with carrier material to produce a single dosage form varies depending on the host being treated and the specific mode of administration.The amount of active ingredient that can be combined with carrier material to produce a single dosage form is generally the amount of compound that produces a therapeutic effect.

[0031] The pharmaceutical preparations provided herein used for carrying out the methods provided herein can be prepared according to any method known in the art for the manufacture of pharmaceuticals.Such drugs can contain sweeteners, flavoring agents, coloring agents and preservatives.The preparations can be mixed with non-toxic pharmaceutically acceptable excipients suitable for manufacturing.The preparations can contain one or more diluents, emulsifiers, preservatives, buffering agents, excipients, etc., and can be provided in the form of liquid, powder, emulsion, lyophilized powder, spray, cream, lotion, controlled release formulation, tablet, pill, gel, on a patch, implant, etc.

[0032] Pharmaceutical preparations for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in suitable dosage amounts. Such carriers allow the medicament to be formulated in unit dosage forms such as tablets, geltabs, pills, powders, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc., suitable for patient ingestion. Pharmaceutical preparations for oral use can also be formulated as solid excipients, and after adding suitable additional compounds, the resulting mixture can be milled as needed, and the mixture of granules can be processed to obtain tablets or dragee cores, if necessary, and formulated. Suitable solid excipients include carbohydrate or protein fillers, such as sugars (including lactose, sucrose, mannitol, or sorbitol); starches derived from corn, wheat, rice, potato, or other plants; celluloses (such as methylcellulose, hydroxypropylmethylcellulose, or sodium carboxymethylcellulose); and gums (including gum arabic and gum tragacanth); and proteins (such as gelatin and collagen). Disintegrants or solubilizers may also be added (e.g., cross-linked polyvinylpyrrolidone, agar, alginic acid, or a salt thereof (e.g., sodium alginate)).

[0033] The dragee cores are provided with a suitable coating (e.g., concentrated sugar solution), which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solution, and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to the tablet or dragee coating for product identification or to characterize the amount of active compound (i.e., dosage). The pharmaceutical preparations provided herein used to carry out the methods provided herein can also be used orally, for example, using push-fit capsules made from gelatin, and soft, sealed capsules made from gelatin and a coating (e.g., glycerol or sorbitol). Push-fit capsules can contain the active agent mixed with a filler or binder (e.g., lactose or starch), a lubricant (e.g., talc or magnesium stearate), and, if necessary, a stabilizer. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol with or without stabilizers.

[0034] Aqueous suspensions may contain an active agent as provided herein (e.g., a human or humanized antibody capable of Fc region-specific binding to the human FcγRI (CD64) receptor but not or substantially not to other human FcγRs, and capable of specifically binding to a cell surface-expressed αvβ3 (avb3) polypeptide, optionally including immune cells) in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents (e.g., sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum arabic), and dispersing or wetting agents (e.g., naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide with long-chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols). Examples of suitable aqueous suspensions include condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides (e.g., polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives (e.g., ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents, and one or more sweetening agents (e.g., sucrose, aspartame, or saccharin). The formulations may be adjusted for osmolality.

[0035] Oil-based medicaments are particularly useful for administering hydrophobic active agents (e.g., anti-αvβ3 (avb3) antibodies) used in carrying out the methods provided herein. Oil-based suspensions can be formulated by suspending the active agent in vegetable oils (e.g., peanut oil, olive oil, sesame oil, or coconut oil), or mineral oils (e.g., liquid paraffin); or mixtures thereof. See, for example, U.S. Pat. No. 5,716,928, which describes the use of essential oils or essential oil components to increase bioavailability and reduce inter- and intra-individual variability of orally administered hydrophobic pharmaceutical compounds (see also U.S. Pat. No. 5,858,401). The oil suspensions can contain thickeners (e.g., beeswax, hard paraffin, or cetyl alcohol). Sweeteners can be added (e.g., glycerol, sorbitol, or sucrose) to provide a palatable oral preparation. These formulations can be preserved by adding antioxidants (e.g., ascorbic acid). As an example of an injectable oil vehicle, see Minto (1997) J. See Pharmacol. Exp. Ther. 281:93-102. The pharmaceutical formulations provided herein may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil or a mineral oil (described above), or a mixture thereof. Suitable emulsifying agents include naturally occurring gums (e.g., acacia gum and tragacanth gum), naturally occurring phosphatides (e.g., soybean lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan monooleate), and condensation products of these partial esters with ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). Emulsions may also contain sweeteners and flavoring agents, as in the formulation of syrups or elixirs. Such formulations may also contain demulcents, preservatives, or coloring agents.

[0036] In practicing the embodiments provided herein, the pharmaceutical compounds may also be administered via intranasal, intravenous (IV), intramuscular, sublingual, intraocular, and intravaginal routes, including suppositories, inhalations, powders, and aerosol formulations (for examples of steroid inhalants, see Rohatagi (1995) J. Clin. Pharmacol. 35:1187-1193; Tjwa (1995) Ann. Allergy Asthma Immunol. 75:107-111). Suppository formulations may be prepared by mixing a drug with a suitable non-irritating excipient that is solid at normal temperatures but liquid at body temperature, thereby melting and releasing the drug in the body. Such materials are cocoa butter and polyethylene glycol.

[0037] In practicing the embodiments provided herein, the pharmaceutical compounds may be delivered transdermally, by topical routes, and may be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.

[0038] In carrying out the embodiments provided herein, the pharmaceutical compound can also be delivered as microspheres for slow release in the body.For example, microspheres can be administered via intradermal injection of drugs that slowly release subcutaneously; see Rao (1995) J. Biomater Sci. Polym. Ed. 7:623-645; as biodegradable and injectable gel formulations (see, for example, Gao (1995) Pharm. Res. 12:857-863 (1995)); or as microspheres for oral administration (see, for example, Eyles (1997) J. Pharm. Pharmacol. 49:669-674).

[0039] In practicing the embodiments provided herein, the pharmaceutical compounds may be administered parenterally, for example, by intravenous (IV) administration or administration into a body cavity or lumen of an organ. These formulations may comprise a solution of the active agent dissolved in a pharmaceutically acceptable carrier. Acceptable vehicles and solvents that may be used include water and Ringer's solution, isotonic sodium chloride. Additionally, sterile, fixed oils may be used as solvents or suspending media. For this purpose, any mild, fixed oil may be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids (e.g., oleic acid) may also be used in the preparation of injectables. These solutions are sterile and generally free of undesirable substances. These formulations may be sterilized by conventional, well-known sterilization techniques. The formulations may contain pharmaceutically acceptable auxiliary substances as needed to maintain appropriate physiological conditions (e.g., pH adjusting and buffering agents, toxicity control agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc.). The concentration of the active agent in these preparations can vary widely and is selected mainly based on the volume, viscosity, body weight, etc. of the fluid, depending on the specific administration mode selected and the patient's needs. For IV administration, the preparation can be a sterile injectable preparation (for example, a sterile injectable aqueous or oily suspension). This suspension can be formulated using appropriate dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a suspension in a non-toxic parenterally acceptable diluent or solvent (for example, a solution of 1,3-butanediol). Administration can be by bolus or continuous infusion (for example, substantially continuously introduced into the blood vessel over a certain period of time).

[0040] The pharmaceutical compounds and formulations provided herein used for carrying out the methods provided herein can be lyophilized.Stable lyophilized formulations comprising the compositions provided herein are also provided, which can be produced by lyophilizing a solution comprising the pharmaceutical agent provided herein and a bulking agent (e.g., mannitol, trehalose, raffinose, and sucrose or a mixture thereof).The process for preparing a stable lyophilized formulation can include lyophilizing a solution of about 2.5 mg / mL protein, about 15 mg / mL sucrose, about 19 mg / mL NaCl, and a sodium citrate buffer solution with a pH greater than 5.5 but less than 6.5.See, for example, US Patent Application No. 20040028670.

[0041] The compositions and preparations provided herein that are used for carrying out the methods provided herein can be delivered by using liposome.By using liposome, the delivery of active agent to target cells in vivo can be focused, especially when liposome surface has specific ligand for target cells or otherwise preferentially directed to specific organs.See for example, U.S. Patent No. 6,063,400; U.S. Patent No. 6,007,839;Al-Muhammed (1996) J. Microencapsul.13:293-306;Chonn (1995) Curr.Opin.Biotechnol.6:698-708;Ostro (1989) Am.J.Hosp.Pharm.46:1576-1587.

[0042] The formulations provided herein used to practice the methods provided herein can be administered for prophylactic and / or therapeutic treatment. In therapeutic applications, the compositions are administered to a subject already suffering from a condition, infection, or disease in an amount sufficient to cure, alleviate, or partially halt the clinical manifestations of the condition, infection, or disease and its complications (a "therapeutically effective amount"). For example, in an alternative embodiment, the pharmaceutical compositions provided herein are administered in an amount sufficient to treat or ameliorate advanced cancer (e.g., a drug-resistant cancer expressing αvβ3 polypeptide on its cell surface) or to kill cancer stem cells (CSCs) expressing αvβ3 polypeptide on their cell surface. The amount of pharmaceutical composition adequate to achieve this is defined as a "therapeutically effective dose." Effective administration schedules and amounts for this use, i.e., "dosing regimens," depend on various factors, including the stage of the disease or condition, the severity of the disease or condition, the patient's general health, physical condition, age, etc. In calculating a patient's dosing regimen, the mode of administration is also taken into consideration.

[0043] The dosing regimen also takes into account pharmacokinetic parameters well known in the art, i.e., rate of absorption, bioavailability, metabolism, clearance, etc., of the active agent (see, e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; see the latest Remington's (supra). The state of the art allows the clinician to determine the dosage regimen for each individual patient, active agent, and disease or condition being treated. Guidelines provided for similar compositions used as pharmaceuticals can be used as guidelines to determine the dosage regimen, i.e., the dosage schedule and dosage levels administered in practicing the methods provided herein are correct and appropriate.

[0044] Single or multiple administrations of the formulations are given, depending on the dosage and frequency, as needed and tolerated by the patient. The formulation should provide a sufficient amount of the active agent to effectively treat, prevent, or ameliorate the condition, disease, or symptom as described herein. For example, exemplary pharmaceutical formulations used to practice the methods provided herein or for oral administration of the compositions provided herein may be in a daily dose of between about 0.1 and 0.5 to about 20, 50, 100, or 1000 μg or more per kilogram of body weight per day. In alternative embodiments, dosages of about 1 mg to about 4 mg per kilogram of patient body weight per day are used. Lower dosages may be used into the bloodstream, a body cavity, or the lumen of an organ, as opposed to oral administration. Substantially higher dosages may be used for topical or oral administration, or administration by powder, spray, or inhalation. Actual methods for preparing parenterally or orally administrable formulations are known or apparent to those skilled in the art and are described in more detail in publications such as Remington's (supra).

[0045] The methods provided herein may further include co-administration with other drugs or pharmaceuticals, e.g., compositions for treating cancer, septic shock, infection, fever, pain, and related symptoms or conditions. For example, the methods and / or compositions and formulations provided herein may be co-formulated and / or co-administered with antibiotics (e.g., antibacterial or bacteriostatic peptides or proteins, particularly those effective against gram-negative bacteria), fluids, cytokines, immunomodulatory agents, anti-inflammatory agents, complement-activating agents (e.g., peptides or proteins containing collagen-like or fibrinogen-like domains (e.g., ficolins), carbohydrate-binding domains, etc., and combinations thereof).

[0046] Antibodies and antigen-binding polypeptides as pharmaceutical compositions In alternative embodiments, compositions and methods comprising the antibodies as provided herein (including antibodies used to practice the methods as provided herein) are also provided. In alternative embodiments, compositions for administering these antibodies and polypeptides are provided.

[0047] In an alternative embodiment, the method includes the use of any polypeptide capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide, whose Fc region has selective affinity for human FcγRI (CD64) on effector cells (e.g., macrophages, neutrophils, and dendritic cells) but no or substantially no affinity for other FcγRs.

[0048] Alternative embodiments may use "humanized" antibodies, which include forms of non-human (e.g., rodent) antibodies that are chimeric antibodies containing minimal sequence (e.g., antigen-binding fragments) derived from non-human immunoglobulin. In alternative embodiments, humanized antibodies are human immunoglobulins in which hypervariable region (HVR) residues of a recipient (e.g., human antibody sequences) are replaced by residues from the hypervariable region (HVR) of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate possessing the desired specificity, affinity, and capacity. In alternative embodiments, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues to improve antigen-binding affinity.

[0049] In alternative embodiments, humanized antibodies may contain residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to improve antibody affinity or functional activity. In alternative embodiments, the humanized antibody may comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable regions correspond to those of a non-human immunoglobulin and all or substantially all of the Ab framework regions are those of a human immunoglobulin sequence.

[0050] In an alternative embodiment, a humanized antibody used to practice the embodiments provided herein may comprise at least a portion of an immunoglobulin constant region (Fc), typically that of or derived from a human immunoglobulin.

[0051] However, in alternative embodiments, fully human antibodies, including human antibodies containing amino acid sequences that correspond to the amino acid sequences of antibodies produced by humans, can also be used to practice the embodiments provided herein. This definition of a human antibody specifically excludes humanized antibodies containing non-human antigen-binding residues.

[0052] In an alternative embodiment, the method comprises producing a humanized antibody (humanized VITAXIN) capable of specifically binding to an αvβ3 (avb3) integrin polypeptide. TM (MedImmune) or MEDI-523, etaracizumab (or etaratuzumab), or MEDI-522, or ABEGRIN TM This includes the use of (including MedImmune).

[0053] In an alternative embodiment, the method involves producing a humanized antibody, e.g., VITAXIN, as described in U.S. Patent Nos. 6,590,079, 7,422,744, and 7,422,745. TM In an alternative embodiment, the antibody used to practice the embodiments as provided herein includes the use of α v The antibody includes an antibody that exhibits selective binding affinity to β3 and comprises a heavy chain polypeptide comprising the variable region amino acid sequence shown in Figure 1A (SEQ ID NO: 1) and a light chain polypeptide comprising the variable region amino acid sequence shown in Figure 1B (SEQ ID NO: 2), or a functional fragment thereof.

[0054] In alternative embodiments, antibodies used to practice the embodiments provided herein include "affinity matured" antibodies, e.g., antibodies that contain one or more changes in one or more hypervariable regions that result in an improvement in the affinity of the antibody for an antigen (e.g., a histone methyl- and / or acetyltransferase) compared to a parent antibody that does not possess those changes. In alternative embodiments, antibodies used to practice the embodiments provided herein are matured antibodies that have nanomolar or even picomolar affinity for a target antigen (e.g., a histone methyl- and / or acetyltransferase). Affinity matured antibodies can be produced by procedures known in the art.

[0055] In an alternative embodiment, the antibody used to carry out the method provided herein is the monoclonal antibody LM609; the monoclonal antibody LM609 is described, for example, in Cheresh et al., J. Biol. Chem. 1987;262(36):17703-11; and U.S. Patent Nos. 5,753,230 and 6,590,079. LM609 is a rodent monoclonal antibody specific for integrin αβ3 (see, for example, Cheresh, DA, Proc. Natl. Acad. Sci. USA 84:6471-6475(1987) and Cheresh et al., J. Biol. Chem. 262:17703-17711(1987)). LM609 was raised against and is reactive with the M21 cell adhesion receptor, now known as integrin αvβ3.

[0056] LM609 inhibits the adhesion of M21 cells to αvβ3 ligands (e.g., vitronectin, fibrinogen, and von Willebrand factor) (Cheresh and Spiro, supra), and inhibits αvβ3-mediated pathologies (e.g., tumor-induced angiogenesis (Brooks et al., 2004)). Cell 79:1157-1164(1994)), granulation tissue development in skin wounds (Clark et al., Am. J. Pathology, 148:1407-1421(1996)), and smooth muscle cell migration such as that occurring during restenosis (Choi et al., J. Vascular Surg., 19:125-134(1994); Jones et al., Proc. Natl. Acad. Sci. 93:2482-2487(1996)).

[0057] In an alternative embodiment, the antibody used to practice the methods as provided herein is α vand a grafted LM609-grafted antibody that exhibits selective binding affinity for β3 and comprises a heavy chain polypeptide variable region amino acid sequence shown in FIG. 1A (SEQ ID NO: 2) and a light chain polypeptide (see FIG. 3) substantially comprising a variable region amino acid sequence having the nucleotide sequence and amino acid sequence shown in SEQ ID NO: 9 and SEQ ID NO: 10, respectively, or a functional fragment thereof.

[0058] Manufacturing Products and Kits Articles of manufacture and kits for practicing the methods as provided herein are provided, said articles of manufacture and kits comprising a human or humanized antibody whose Fc region is capable of specifically binding to a cell surface-expressed αvβ3 (avb3) polypeptide that has selective affinity for human FcγRI (CD64) but no or substantially no affinity for other human FcγRs; and / or comprising macrophages, neutrophils, and dendritic cells; and optionally further comprising instructions for practicing the methods as provided herein.

[0059] Any of the above aspects and embodiments may be combined with any other aspect or embodiment as disclosed herein in the Summary and / or Detailed Description sections.

[0060] As used in this specification and the claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0061] Unless specifically stated or apparent from the context, as used herein, the term "or" is understood to be inclusive and to cover both "or" and "and."

[0062] Unless otherwise specified or obvious from the context, when used herein, the term "about" is understood to be within the range normally accepted in the art (for example, within 2 standard deviations of the mean). About (use of the term "about") can be understood as within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values ​​provided herein are modified by the term "about".

[0063] Unless specifically stated or obvious from the context, as used herein, the terms "substantially all," "substantially a majority of," "substantially all of," or "the majority of" encompass at least about 90%, 95%, 97%, 98%, 99%, or 99.5%, or more, of the referenced amount of the composition.

[0064] The entirety of each patent, patent application, publication, and document referred to herein is incorporated herein by reference. Citation of the above patents, patent applications, publications, and documents is not an admission that any of the foregoing is relevant prior art, nor does it constitute any admission as to the content or date of such publications or documents. The incorporation by reference of these documents alone should not be construed as an assertion or admission that any portion of the contents of any document is to be considered essential to satisfy the statutory disclosure requirements of any country or region for patent applications. Nevertheless, the right is reserved to rely on any such documents, where appropriate, to provide material deemed essential to the claimed subject matter by an examining authority or court.

[0065] Modifications may be made to the above without departing from the basic aspects of the invention. While the present invention has been described in substantial detail with reference to one or more specific embodiments, those skilled in the art will recognize that changes may be made to the embodiments specifically disclosed in this application, and that these changes and improvements will still be within the scope and spirit of the invention. The invention as properly illustrated herein may be practiced in the absence of any element not specifically disclosed herein. Thus, for example, in each instance herein, any of the terms "comprise," "consist essentially of," and "consist" may be substituted for either of the other two terms. Thus, the terms and expressions used are used in a descriptive sense, not in a limiting sense, and equivalents of the features shown and described, or portions thereof, are not excluded, and it is recognized that various modifications are possible within the scope of the invention.

[0066] Although a number of embodiments of the invention have been described, it will be understood that various modifications can be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims. In certain embodiments, for example, the following are provided: (Item 1) - Treating or ameliorating cancer, where appropriate, advanced or drug-resistant cancer; or - Killing cancer stem cells (CSCs), wherein the cancer, the advanced cancer, the drug-resistant cancer, or the CSC expresses an αvβ3 polypeptide on their cell surface; The method comprises administering to an individual in need thereof a human or humanized antibody capable of Fc region-specific binding to the human FcγRI (CD64) receptor but no or substantially no binding to other human FcγRs, and capable of specific binding to cell surface-expressed αvβ3 (avb3) polypeptide; Optionally, the human FcγRI (CD64) receptor is expressed on the surface of human macrophages, neutrophils and / or dendritic cells; This method induces an antibody-dependent cell-mediated cytotoxicity (ADCC) response or reaction against the cells of the advanced or drug-resistant cancer, or CSCs. (Item 2) 2. The method of claim 1, wherein the human or humanized antibody comprises monoclonal antibody (mAb) LM609 (MedImmune), or a mAb having ATCC accession number HB9537, or a mAb as described in U.S. Pat. No. 5,753,230. (Item 3) The human or humanized antibody is VITAXIN TM (MedImmune) or MEDI-523. (Item 4) The human or humanized antibody may be etaracizumab (or etaratuzumab), MEDI-522, or ABEGRIN TM (MedImmune). (Item 5) The method of any of the preceding items, further comprising administering to the individual in need thereof an additional cancer therapeutic agent or treatment, wherein optionally the additional cancer therapeutic agent comprises paclitaxel. (Item 6) The method of any of the preceding items, wherein the human antibody or humanized antibody is administered to the individual in need thereof at a dose of between about 1 and about 8 mg / kg, or between about 0.5 and about 12 mg / kg. (Item 7) The method of any of the preceding items, wherein the human or humanized antibody is administered intravenously (IV), intrathecally, sublingually, rectally, intravaginally, subcutaneously, orally, or intramuscularly (IM), or is injected or placed near, proximal to, or into a cancer or tumor (optionally a solid tumor), or an advanced or drug-resistant cancer, or CSC in situ, or is administered by in situ placement or insertion of an implant comprising the human or humanized antibody. (Item 8) The additional cancer therapeutic agent or treatment may be abagovomab, adecatumumab, afutuzumab, alemtuzumab, altumomab, amatuximab, anatumomab, arcitumomab, bavituximab, bectumomab, bevacizumab, bivatuzumab, blinatumomab, brentuximab, cantuzumab, catumaxomab, cetuximab, sitatuzumab, cixutumumab, clivatuzumab, conatumumab, daratumumab, drozitumab, durigotumab, dusigitumab, , detumomab, dacetuzumab, dalotuzumab, ecromeximab, elotuzumab, ensituximab, ertumaxomab, farletuzumab, ficlatuzumab, figitumumab, framvotumab, futuximab, ganitumab, gemtuzumab, girentuximab, glenbatumumab, ibritumomab, igovomab, imagatuzumab, indatuximab, inotuzumab, intetumumab, ipilimumab, iratumumab, labetuzumab, lexatumumab, lintuzumab , lorvotuzumab, lucatumumab, mapatumumab, matuzumab, milatuzumab, minletumomab, mitumomab, moxetumomab, narunatumumab, naptumomab, necitumumab, nimotuzumab, nofetumomab, ocaratuzumab, ofatumumab, olaratumumab, onartuzumab, oportuzumab, oregovomab, panitumumab, palsatuzumab, patritumab, pemtumomab, pertuzumab, pintumomab, pritumumab, racotumomab, radletuzumab, rilo 6. The method of item 5, wherein the antibody comprises or is an antibody selected from the group consisting of: rituximab, lobatumumab, satumomab, sibrotuzumab, siltuximab, simtuzumab, solitomab, tacatuzumab, taplitumomab, tenatumomab, teprotumumab, tigatuzumab, tositumomab, trastuzumab, tucotuzumab, ublituximab, veltuzumab, borsetuzumab, votumumab, zalutumumab, and / or any combination thereof. (Item 9) The additional cancer therapeutic agent or cancer treatment comprises a growth factor inhibitor, wherein optionally the growth factor inhibitor is a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM, mitotic toxin, paclitaxel, taxol, abraxane TM , erlotinib, TARCEVA TM , lapatinib, TYKERB TM , cetuximab, ERBITUX TM 6. The method of item 5, comprising a PD-1 inhibitor, a PD-L1 inhibitor and / or an insulin growth factor inhibitor. (Item 10) A plurality of said human or humanized antibodies may be dehumanized prior to administration to said individual in need thereof. pre-incubated ex vivo with phage, neutrophil, monocyte and / or dendritic cell, The method of any of the preceding items, wherein optionally the human macrophages, neutrophils, monocytes and / or dendritic cells are activated human macrophages, neutrophils, monocytes and / or dendritic cells, optionally the dendritic cells are activated as set forth in U.S. Pat. No. 10,023,841, optionally the monocytes or dendritic cells are antigen-loaded monocytes or dendritic cells, optionally the monocytes or dendritic cells are antigen-loaded, activated monocytes or dendritic cells. (Item 11) - Treating or ameliorating cancer or tumors, or advanced or drug-resistant cancers, or - Killing cancer stem cells (CSCs), 1. Use of a human or humanized antibody capable of Fc region-specific binding to human FcγRI (CD64) receptor but not or substantially not to other human FcγRs, and capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide, for wherein said cancer or tumor, or said advanced cancer, drug-resistant cancer or CSC, expresses αvβ3 polypeptide on their cell surface. (Item 12) - Treating or ameliorating cancer or tumors, or advanced or drug-resistant cancers, or - Killing cancer stem cells (CSCs), A human or humanized antibody capable of Fc region-specific binding to human FcγRI (CD64) receptor but not or substantially not to other human FcγRs, and capable of specifically binding to cell surface-expressed αvβ3 (avb3) polypeptide, for use in A human or humanized antibody, wherein the cancer or tumor, or the advanced cancer, drug-resistant cancer, or CSC expresses an αvβ3 polypeptide on their cell surface.

Claims

[Claim 1] The invention described in the specification.

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