Compositions and methods for targeting and killing alpha-vbeta-3 (alphavbeta3) positive cancer stem cells (csc) and treating drug resistant and metastatic cancers

Abstract

The present invention provides compositions and methods for treating or ameliorating advanced cancers, such as drug-resistant or metastatic cancers, or killing cancer stem cells expressing an alpha v beta 3 polypeptide on its cell surface by using a human antibody or a humanized antibody, and the human antibody or humanized antibody is capable of specifically binding to an [alpha] v [beta] 3 (avb3) polypeptide expressed on the cell surface of effector cells such as macrophages, neutrophils and dendritic cells, the Fc region of which has selective affinity for human Fc [gamma] RI (CD64) but does not have selective affinity for other Fc [gamma] R. By administering these antibodies to an individual in need thereof, these human antibodies or humanized antibodies can treat, ameliorate, or slow the progression of advanced cancers or drug resistant cancers or cancers caused, initiated, or maintained by advanced cancers or drug resistant cancer cells or cancer stem cells.

Description

[0001] This application is a divisional application of an invention patent application that entered the Chinese national phase on January 12, 2022, with application number 202080050743.3, filing date July 31, 2020, and invention name “Compositions and methods for targeting and killing α-vβ-3 (αvβ3) positive cancer stem cells (CSCs) and treating drug-resistant and metastatic cancers”.

[0002] Related Applications

[0003] This Patent Cooperation Treaty (PCT) international application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62 / 882,296, filed on August 2, 2019. The foregoing application is expressly incorporated herein by reference in its entirety for all purposes. All publications, patents, and patent applications cited herein are hereby expressly incorporated herein by reference for all purposes. Technical Field

[0004] The present invention relates generally to immunology and oncology. In alternative embodiments, compositions and methods for treating or improving advanced cancers such as drug-resistant or metastatic cancers or killing cancer stem cells (CSCs) are provided by using human antibodies or humanized antibodies, wherein the advanced cancers, drug-resistant or metastatic cancers, cancer stem cells express αvβ3 polypeptides on their cell surfaces, and the human antibodies or humanized antibodies are capable of specifically binding to αvβ3 (avb3) polypeptides expressed on the cell surfaces of effector cells such as macrophages, neutrophils, and dendritic cells, and their Fc regions have selective affinity for human FcγRI (CD64), but do not have selective affinity for other FcγRs. By administering these antibodies to individuals in need, these human antibodies or humanized antibodies are capable of treating, improving, or slowing down the development of advanced cancers or drug-resistant cancers or cancers caused or initiated or maintained by advanced cancers or drug-resistant cancer cells or cancer stem cells (CSCs). In alternative embodiments, the administered human or humanized antibody induces an antibody-dependent cell-mediated cytotoxicity (ADCC) response against advanced cancer or drug-resistant cancer cells or CSCs. Background Art

[0005] Antibodies utilize effector cells such as macrophages, natural killer cells, dendritic cells, and neutrophils to induce antibody-dependent cell-mediated cytotoxicity (ADCC) against target cells. In order to utilize these effector cells, the Fc of the antibody needs to have affinity for Fcγ receptors (FcγR) on the effector cells. Summary of the invention

[0006] In alternative embodiments, methods are provided for:

[0007] - Treat or ameliorate cancer, optionally advanced cancer or drug-resistant cancer, or

[0008] - Kill cancer stem cells (CSC),

[0009] wherein the cancer, advanced cancer, drug-resistant cancer or CSC expresses the αvβ3 polypeptide on its cell surface,

[0010] The method comprises administering to an individual in need thereof a human antibody or a humanized antibody, which can specifically bind to a human FcγRI (CD64) receptor in its Fc region but does not bind or substantially bind to other human FcγRs, and can specifically bind to an αvβ3 (avb3) polypeptide expressed on a cell surface,

[0011] wherein optionally human FcγRI (CD64) receptor is expressed on the surface of human macrophages, neutrophils and / or dendritic cells,

[0012] Thereby, an antibody-dependent cell-mediated cytotoxicity (ADCC) response or reaction is induced against advanced cancer or drug-resistant cancer cells or CSCs.

[0013] In alternative embodiments of the methods provided herein:

[0014] - the human antibody or humanized antibody comprises monoclonal antibody (mAb) LM609 (MedImmune) or the monoclonal antibody with ATCC accession number HB9537 or the monoclonal antibody as described in U.S. Patent Serial No. (USSN) 5,753,230;

[0015] - The human antibody or humanized antibody includes VITAXIN TM (MedImmune) or MEDI-523;

[0016] - The human antibody or humanized antibody includes etaracizumab or etaratuzumab or MEDI-522 or ABEGRIN TM (MedImmune);

[0017] - the method further comprises administering to the individual in need thereof an additional cancer therapeutic agent or therapy, wherein optionally the additional cancer therapeutic agent comprises paclitaxel;

[0018] - administering the human antibody or humanized antibody to a subject in need thereof at a dose of about 1 mg / kg to about 8 mg / kg or about 0.5 mg / kg to about 12 mg / kg;

[0019] - the human antibody or humanized antibody is administered intravenously (IV), intrathecally, sublingually, rectally, intravaginally, subcutaneously, or intramuscularly (IM), or injected or placed in situ near, adjacent to, or within a cancer or tumor (e.g., a solid tumor) or an advanced cancer or a drug-resistant cancer or a CSC, or administered by in situ placement or insertion of an implant comprising the human antibody or humanized antibody;

[0020] - the additional cancer therapeutic agent or therapy comprises an antibody selected from the group consisting of abavolumab, adelimumab, afutuzumab, alemtuzumab, atumomab, amatumomab, maanatumomab, acitumomab, baviximab, betumomab, bevacizumab, bivacizumab, blinatumomab, brentuximab, cantuzumab, catumaxomab, cetuximab, situzumab, citumumab, clituzumab, canatumumab , daratumumab, trazitumab, dulitumab, dustuzumab, demostatumab, dasitumumab, dalozumab, emeticizumab, elotuzumab, entuximab, ertuinomab, fatuinomab, felatuzumab, fentolimumab, falotuzumab, vortuximab, ganituzumab, gemtuzumab, gemtuximab, gebatumumab, ibritumomab tiuxetan, igovotumab, imaclizumab, intuximab, intuzumab, intuzumab, ipilimumab, Itolumab, labetuzumab, lexatumumab, lintuzumab, lovotuzumab, rucamumab, mapatumumab, matuzumab, miratumumab, mitumumab, mitumumab, moxitumomab, nanatumab, naptuzumab, nexitumomab, nimotuzumab, novatumumab, okatumumab, ofatumumab, olaratumab, onatumumab, moxotuzumab, ogavuzumab, panitumumab, pasatuzumab, pertuzumab, pemuzumab , pertuzumab, pintumomab, pligumab, rantumomab, rantrastuzumab, rituximab, rituximab, rotuximab, satumomab, sirolimus, silotuzumab, sintuzumab, solitotumab, talizumab, talizumab, talitumomab, tetuzumab, tetuzumab, tegazumab, tositumomab, trastuzumab, tukavicuzumab, utiluxamab, veltuzumab, worse tuzumab, vortumomab, zalutumumab, and / or any combination thereof;

[0021] - the additional cancer therapeutic agent or therapy comprises a growth factor inhibitor, wherein optionally the growth factor inhibitor comprises a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM , mitotic poisons, paclitaxel, taxol, ABRAXANE TM , erlotinib, TARCEVA TM, Lapatinib, TYKERB TM , Cetuximab, ERBITUX TM , PD-1 inhibitors, PD-L1 inhibitors and / or insulin growth factor inhibitors; and / or

[0022] - Pre-incubating the plurality of human or humanized antibodies with human macrophages, neutrophils, monocytes and / or dendritic cells in vitro prior to administration to an individual in need thereof, wherein optionally the human macrophages, neutrophils, monocytes and / or dendritic cells are activated human macrophages, neutrophils, monocytes and / or dendritic cells, and optionally the dendritic cells or monocytes are activated as described in USPN 10,023,841.

[0023] In an alternative embodiment, a use of a human antibody or a humanized antibody is provided, wherein the human antibody or the humanized antibody can specifically bind to the human FcγRI (CD64) receptor in the Fc region but does not bind or substantially bind to other human FcγRs, and can specifically bind to the αvβ3 (avb3) polypeptide expressed on the surface of a cell, and the use is used for

[0024] - Treat or ameliorate cancer or tumors, or advanced cancer or drug-resistant cancer, or

[0025] - Kill cancer stem cells (CSC),

[0026] Wherein the cancer or tumor, or advanced cancer, drug-resistant cancer or CSC expresses the αvβ3 polypeptide on its cell surface.

[0027] In an alternative embodiment, a human antibody or a humanized antibody is provided, which can specifically bind to the human FcγRI (CD64) receptor in the Fc region but does not bind or substantially binds to other human FcγRs, and can specifically bind to the αvβ3 (avb3) polypeptide expressed on the surface of a cell, and the human antibody or the humanized antibody is used for:

[0028] - Treat or ameliorate cancer or tumors, or advanced cancer or drug-resistant cancer, or

[0029] - Kill cancer stem cells (CSC),

[0030] Wherein the cancer or tumor, or advanced cancer, drug-resistant cancer or CSC expresses the αvβ3 polypeptide on its cell surface.

[0031] The details of one or more exemplary embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and claims.

[0032] All publications, patents, and patent applications cited herein are hereby expressly incorporated by reference for all purposes. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The drawings presented herein are only for purposes of illustrating exemplary embodiments provided herein and are not meant to limit the scope of the present invention as encompassed by the claims.

[0034] Figure 1A-1B The antibody VITAXIN was shown TM The nucleotide and amino acid sequences of the variable regions are: Figure 1A The nucleotide sequence and amino acid sequence of the heavy chain variable region are shown (SEQ ID NO: 1 and SEQ ID NO: 2, respectively) and Figure 1B The nucleotide and amino acid sequences of the light chain variable region are shown (SEQ ID NO: 3 and SEQ ID NO: 4, respectively).

[0035] Figure 2A-2B The nucleotide and amino acid sequences of the variable regions of monoclonal antibody LM609 are shown; Figure 2A shows the nucleotide sequence and amino acid sequence of the heavy chain variable region of LM609 (SEQ ID NO: 5 and SEQ ID NO: 6, respectively), which variable region extends from amino acid Glu1 to Ala117; and Figure 2B The nucleotide sequence and amino acid sequence of the LM609 light chain variable region are shown (SEQ ID NO: 7 and SEQ ID NO: 8, respectively).

[0036] Figure 3 Light chain polypeptides (for use with Figure 1A The light chain polypeptide comprises a variable region amino acid sequence having a nucleotide sequence and an amino acid sequence as shown in SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

[0037] Like reference numbers in the various drawings represent the same elements. DETAILED DESCRIPTION

[0038] In alternative embodiments, compositions and methods for treating or improving cancer or tumors, for example, advanced cancers such as drug-resistant or metastatic cancers, or killing cancer stem cells (CSCs) by using (by administering) human antibodies or humanized antibodies are provided, wherein the cancer or tumor, advanced cancer, drug-resistant or metastatic cancer, cancer stem cells express αvβ3 polypeptides on their cell surfaces, and the human antibodies or humanized antibodies are capable of specifically binding to αvβ3 (avb3) polypeptides expressed on the cell surfaces of effector cells such as macrophages, neutrophils, and dendritic cells, and their Fc regions have selective affinity for human FcγRI (CD64), but do not have or substantially do not have selective affinity for other FcγRs. By administering these antibodies to individuals in need or practicing the methods provided by the present invention, these human antibodies or humanized antibodies can treat, improve cancer or tumors, advanced cancers or drug-resistant cancers, or cancers caused or initiated or maintained by advanced cancer or drug-resistant cancer cells or cancer stem cells (CSCs), or slow down their development. In alternative embodiments, the administered human or humanized antibody induces an antibody-dependent cell-mediated cytotoxicity (ADCC) response against advanced cancer or drug-resistant cancer cells or CSCs.

[0039] Pharmaceutical compositions and preparations

[0040] In alternative embodiments, pharmaceutical compositions and preparations for treating or ameliorating advanced cancers such as drug-resistant cancers or killing cancer stem cells (CSCs) are provided, for example, pharmaceutical compositions and preparations comprising human antibodies or humanized antibodies capable of specifically binding to αvβ3 (avb3) polypeptides expressed on the cell surface of effector cells such as macrophages, neutrophils and dendritic cells, whose Fc region has selective affinity for human FcγRI (CD64) but has no or substantially no selective affinity for other FcγRs, and methods, wherein advanced cancers, drug-resistant cancers, cancer stem cells express the 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, such as macrophages, neutrophils, monocytes and / or dendritic cells or activated forms thereof, optionally comprising macrophages, neutrophils, monocytes and / or dendritic cells that have been pre-incubated in vitro with human antibodies or humanized antibodies that are capable of specifically binding to αvβ3 (avb3) polypeptides expressed on the surface of cells.

[0041] In alternative embodiments, compositions provided herein and compositions for implementing methods provided herein are formulated using pharmaceutically acceptable carriers. In alternative embodiments, pharmaceutical compositions for implementing methods provided herein can be administered parenterally, topically, orally, intrathecally, sublingually, rectal, intravaginally, subcutaneously or intramuscularly (IM) or by any form of topical administration, such as by aerosol or transdermal administration. Pharmaceutical compositions can be formulated in any manner and can be administered in various unit dosage forms, depending on the condition or disease and degree of disease, the basic medical condition of each patient, the final preferred method of administration, etc. Details about preparations and administration techniques are described in detail in scientific and patent literature, see, for example, Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's") latest edition.

[0042] The therapeutic agents provided herein, such as therapeutic agents including anti-αvβ3 (avb3) antibodies, and antibodies for practicing the methods provided herein, can be administered alone or as a component of a pharmaceutical formulation (composition), or administered simultaneously with, before and / or after another active agent, such as a growth factor inhibitor, wherein optionally, the growth factor inhibitor includes a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM , mitotic poisons, paclitaxel, taxol, ABRAXANE TM , erlotinib, TARCEVA TM , Lapatinib, TYKERB TM , Cetuximab, ERBITUX TM , PD-1 inhibitors, PD-L1 inhibitors and / or insulin growth factor inhibitors.

[0043] Pharmaceutical compositions and formulations, for example those comprising anti-αvβ3 (avb3) antibodies, can be formulated for human or veterinary administration in any convenient manner.

[0044] Wetting agents, emulsifiers and lubricants, such as 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 compositions.

[0045] Formulations of the compositions provided herein and for practicing the methods provided herein include those suitable for oral, nasal, topical, parenteral, rectal, subcutaneous, sublingual, intraocular, intramuscular, intrathecal and / or intravaginal administration.

[0046] These preparations can be conveniently presented in unit dosage form and can be prepared by any method well known in the pharmaceutical art. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally that amount of the compound that produces a therapeutic effect.

[0047] The pharmaceutical preparations provided herein and the pharmaceutical preparations for implementing the methods provided herein can be prepared according to any method known in the pharmaceutical field. Such medicines can contain sweeteners, flavoring agents, coloring agents and preservatives. The preparations can be mixed with nontoxic pharmaceutically acceptable excipients suitable for manufacture. The preparations can include one or more diluents, emulsifiers, preservatives, buffers, excipients, etc., and can be provided in forms such as liquids, powders, emulsions, freeze-dried powders, sprays, creams, lotions, controlled release preparations, tablets, pills, gels, patches, implants.

[0048] Pharmaceutical preparations for oral administration can be formulated with appropriate and suitable dosages using pharmaceutically acceptable carriers well known in the art. Such carriers enable the drug to be formulated into unit doses of tablets, gel tablets, pills, powders, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc. suitable for patient intake. Oral pharmaceutical preparations can be formulated into solid excipients, optionally grinding the resulting mixture, and if necessary, processing the granular mixture after adding a suitable additional compound to obtain tablets or dragee cores. Suitable solid excipients are carbohydrate fillers or protein fillers, including, for example, sugars, including lactose, sucrose, mannitol or sorbitol; starch from corn, wheat, rice, potatoes or other plants; cellulose, such as methylcellulose, hydroxypropyl methylcellulose or sodium carboxymethylcellulose; and gums, including gum arabic and tragacanth; and proteins, such as gelatin and collagen. Disintegrants or solubilizers such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or its salts (such as sodium alginate) can be added.

[0049] The sugar-coated pill core has a suitable coating, such as a concentrated sugar solution, which can also contain gum arabic, talcum, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol and / or titanium dioxide, lacquer solution and a suitable organic solvent or solvent mixture. Dye or pigment can be added to tablets or sugar-coated pill coatings for identifying products or characterizing the amount (i.e. dosage) of active compounds. The pharmaceutical preparations provided herein and the pharmaceutical preparations for implementing the methods provided herein can also be used orally, for example, using push-in capsules made of gelatin and soft sealed capsules made of gelatin and coatings such as glycerol or sorbitol. Push-in capsules can contain an activating agent mixed with a filler or adhesive (such as lactose or starch), a lubricant (such as talcum or magnesium stearate) and an optional stabilizer. In soft capsules, these activating agents can be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin or liquid polyethylene glycol containing or not containing a stabilizer.

[0050] The aqueous suspension may contain an active agent as provided herein, such as a human antibody or humanized antibody that is capable of specifically binding to the human FcγRI (CD64) receptor at its Fc region but does not bind or substantially binds to other human FcγRs and is capable of specifically binding to the αvβ3 (avb3) polypeptide expressed on the surface of cells (optionally including immune cells), in admixture with excipients suitable for preparing aqueous suspensions. Such excipients include suspending agents (such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth and gum arabic) and dispersants or wetting agents, such as naturally occurring phosphatides (such as lecithin), condensation products of alkylene oxide and fatty acids (such as polyoxyethylene stearate), condensation products of ethylene oxide and long-chain fatty alcohols (such as heptadecaethyleneoxy hexadecyl alcohol), condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol (such as polyoxyethylene sorbitol monooleate) or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol anhydrides (such as polyoxyethylene sorbitan monooleate). The aqueous suspension can also contain one or more preservatives (such as ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavors and one or more sweeteners (such as sucrose, aspartame or saccharin). The osmotic pressure of the preparation can be adjusted.

[0051] Oil-based drugs are particularly suitable for administering hydrophobic active agents (e.g., anti-αvβ3 (avb3) antibodies) for implementing the methods provided herein. Oil-based suspensions can be prepared by suspending the active agent in a vegetable oil (such as peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (such as liquid paraffin) or a mixture of these oils. See, for example, U.S. Pat. No. 5,716,928 (see also U.S. Pat. No. 5,858,401), which describes the use of essential oils or essential oil components to improve the bioavailability of orally administered hydrophobic drug compounds and reduce the variability of the compound between and within individuals. The oil suspension may contain a thickener, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners may be added to provide a palatable oral preparation, such as glycerol, sorbitol, or sucrose. These preparations may be preserved by the addition of antioxidants (such as ascorbic acid). As an example of injectable oily solvents, see Minto (1997), "Journal of Pharmacology and Experimental Therapeutics" (J.Pharmacol.Exp.Ther.), Vol. 281, pp. 93-102. The pharmaceutical preparations provided herein can also be in the form of oil-in-water emulsions. The oil phase can be a vegetable oil or mineral oil as described above or a mixture of these oils. Suitable emulsifiers include naturally occurring gums (such as gum arabic and tragacanth), naturally occurring phospholipids (such as soybean lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (such as sorbitan monooleate) and condensation products of these partial esters with ethylene oxide (such as polyoxyethylene sorbitan monooleate). The emulsifier can also contain sweeteners and flavors, such as sweeteners and flavors in syrup and elixir preparations. Such preparations can also contain a demulcent, a preservative or a coloring agent.

[0052] In practicing the embodiments provided herein, the drug compound can also be administered by intranasal, intravenous (IV), intramuscular, sublingual, intraocular and intravaginal routes, including suppositories, insufflations, powders and aerosol formulations (for steroid inhaler examples, see Rohatagi (1995), Journal of Clinical Pharmacology (J. Clin. Pharmacol.), Vol. 35, pp. 1187-1193; Tjwa (1995), Ann. Allergy Asthma Immunol., Vol. 75, pp. 107-111). Suppository formulations can be prepared by mixing the drug with a suitable non-irritating excipient, which is solid at room temperature but liquid at body temperature and will therefore melt in the body to release the drug. Such materials are cocoa butter and polyethylene glycol.

[0053] In practicing the embodiments provided herein, pharmaceutical compounds formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols can be delivered transdermally via a topical route.

[0054] In the implementation of the embodiments provided herein, the drug compound can also be delivered as a microsphere to slowly release in vivo. For example, slow subcutaneous release microspheres can be administered by intradermal injection; see Rao (1995), "Journal of Biomaterials Science: Polymer Edition" (J.Biomater Sci.Polym.Ed.), Vol. 7, pp. 623-645; as a biodegradable and injectable gel formulation, see, for example, Gao (1995), "Pharmaceutical Research" (Pharm.Res.) Vol. 12, pp. 857-863, 1995; or as a microsphere for oral administration, see, for example, Eyles (1997), "Journal of Pharmacy and Pharmacology" (J.Pharm.Pharmacol.), Vol. 49, pp. 669-674.

[0055] In the implementation of the embodiments provided herein, the drug compound can be administered parenterally, such as by intravenous (IV) administration or by administration into a body cavity or organ lumen. These preparations can include an active agent solution dissolved in a pharmaceutically acceptable carrier. Acceptable solvents and solvents that can be used are water and Ringer's solution (a kind of isotonic sodium chloride). In addition, sterile fixed oils can be used as solvents or suspension media. For this reason, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can also be used to prepare injectable preparations. These solutions are sterile and generally do not contain undesirable substances. These preparations can be sterilized by conventional, well-known sterilization techniques. These preparations can contain pharmaceutically acceptable auxiliary substances required for close physiological conditions, such as pH regulators and pH buffers and toxicity regulators, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc. The concentration of the active agent in these preparations can vary widely, and will be mainly based on fluid volume, viscosity, body weight, etc., according to the selected specific mode of administration and the needs of the patient. For IV administration, the preparation can be a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. The suspension can be prepared using those suitable dispersants or wetting agents and suspending agents. The sterile injectable preparation can also be a suspension in a non-toxic parenteral acceptable diluent or solvent, such as 1,3-butanediol solution. Administration can be by push injection or continuous infusion (e.g., introduced into a blood vessel substantially uninterruptedly over a specified period of time).

[0056] The pharmaceutical compounds and preparations provided herein and the pharmaceutical compounds and preparations for implementing the methods provided herein can be lyophilized. Also provided are stable lyophilized preparations including compositions provided herein, which can be prepared by lyophilizing a solution including a medicine provided herein and a filler (e.g., mannitol, trehalose, raffinose and sucrose or a mixture thereof). A method for preparing a stable lyophilized preparation can include lyophilizing a solution of a sodium citrate buffer solution of about 2.5 mg / mL protein, about 15 mg / mL sucrose, about 19 mg / mL NaCl and pH greater than 5.5 but less than 6.5. See, for example, U.S. Patent Application No. 20040028670.

[0057] Compositions and preparations provided herein and compositions and preparations for implementing the methods provided herein can be delivered by using liposomes. By using liposomes, especially when the liposome surface carries a ligand specific to the target cell or preferentially targets a specific organ, people can focus on delivering the active agent to the target cell in the body. See, for example, U.S. Patent No. 6,063,400; U.S. Patent No. 6,007,839; Al-Muhammed (1996), Journal of Microencapsulation (J. Microencapsul.), Vol. 13, pp. 293-306; Chonn (1995), Contemporary Biotechnology Review (Curr. Opin. Biotechnol.), Vol. 6, pp. 698-708; Ostro (1989), Am. J. Hosp. Pharm., Vol. 46, pp. 1576-1587.

[0058] The formulations provided herein and the formulations for implementing the methods provided herein can be used for preventive treatment administration and / or therapeutic treatment administration. In therapeutic applications, the composition is administered to a subject already suffering from a condition, infection or disease in an amount sufficient to cure, alleviate or partially prevent the clinical manifestations of the condition, infection or disease and its complications ("therapeutically effective amount"). For example, in alternative embodiments, the pharmaceutical compositions provided herein are administered in an amount sufficient to treat or improve advanced cancers (such as drug-resistant cancers expressing αvβ3 polypeptides on their cell surfaces) or to kill cancer stem cells (CSCs) expressing αvβ3 polypeptides on their cell surfaces. The amount of the pharmaceutical composition sufficient to achieve this is defined as a "therapeutically effective dose". The dosage regimen and amount (i.e., "dosage regimen") effective for this use will depend on a variety of factors, including the stage of the disease or condition, the severity of the disease or condition, the general health status of the patient, the patient's physical condition, age, etc. When calculating the patient's dosage regimen, the mode of administration is also taken into account.

[0059] The dosage regimen also takes into account pharmacokinetic parameters well known in the art, i.e., absorption rate, bioavailability, metabolism rate, clearance rate, etc. of the active agent (see, e.g., Hidalgo-Aragones (1996), J. Steroid Biochemistry and Molecular Biology). (1996), Pharmazie, Vol. 51, pp. 337-341; Fotherby (1996), Contraception, Vol. 54, pp. 59-69; Johnson (1995), J. Pharm. Sci., Vol. 84, pp. 1144-1146; Rohatagi (1995), Pharmazie, Vol. 50, pp. 610-613; Brophy (1983), Eur. J. Clin. Pharmacol., Vol. 24, pp. 103-108; the latest by Remington, supra). The prior art allows the clinician to determine the dosage regimen for each individual patient, active agent, and disease or condition being treated. The guidance provided for similar compositions used as pharmaceuticals can be used to determine whether the dosing regimen (ie, dosage schedule and dosage level) for practicing the methods provided herein is correct and appropriate.

[0060] According to the dosage and frequency required and tolerated by the patient, single or multiple administrations of the preparation can be given. The preparation should provide a sufficient amount of active agent to effectively treat, prevent or improve the condition, disease or symptom as described herein. For example, an exemplary pharmaceutical preparation for oral administration of a composition provided herein or a composition for implementing the method provided herein can be administered at a daily dose of about 0.1 μg to 0.5 μg, to about 20 μg, about 50 μg, about 100 μg or about 1000 μg or more μg per kg body weight per day. In an alternative embodiment, the dosage used is about 1 mg to about 4 mg per kg body weight per day per patient. Compared with oral administration, a lower dose can be used to enter the bloodstream, enter the body cavity or enter the lumen of an organ. Substantially higher doses can be used in topical or oral administration or by powder, spray or inhalation administration. The actual method of preparing parenteral or non-parenteral administration preparations is known or obvious to those skilled in the art, and is described in more detail in the above-mentioned publications such as Remington.

[0061] The methods provided herein may also include co-administration with other drugs or medicines, for example, compositions for treating cancer, septic shock, infection, fever, pain and related symptoms or conditions. For example, the methods and / or compositions and preparations provided herein may be co-formulated and / or co-administered with antibiotics (e.g., antibacterial or antibacterial peptides or proteins), particularly antibiotics effective against gram-negative bacteria, fluids, cytokines, immunomodulators, anti-inflammatory agents, complement activators (such as, peptides or proteins including collagen-like domains or fibrinogen-like domains (e.g., fibrinolysin) or carbohydrate-binding domains), etc. and combinations thereof.

[0062] Antibodies and antigen-binding polypeptides as pharmaceutical compositions

[0063] In alternative embodiments, compositions and methods comprising the antibodies provided herein are also provided, including antibodies for practicing the methods provided herein. In alternative embodiments, compositions for administering these antibodies and polypeptides are provided.

[0064] In alternative embodiments, the method comprises using any polypeptide capable of specifically binding to an αvβ3 (avb3) polypeptide expressed on the cell surface of effector cells such as macrophages, neutrophils and dendritic cells, wherein the Fc region of the polypeptide has selective affinity for human FcγRI (CD64) but has no or substantially no selective affinity for other FcγRs.

[0065] Alternative embodiments can use "humanized" antibodies, including the form of non-human (e.g., mouse) antibodies, which are chimeric antibodies including a minimal sequence (e.g., antigen binding fragment) derived from a non-human immunoglobulin. In alternative embodiments, humanized antibodies are human immunoglobulins, wherein residues from receptor hypervariable regions (HVRs) (e.g., human antibody sequences) are replaced by residues from hypervariable regions (HVRs) of non-human species (donor antibodies), wherein non-human species such as mice, rats, rabbits, or non-human primates, the hypervariable regions of these species have desired specificity, affinity, and ability. In alternative embodiments, the framework region (FR) residues of human immunoglobulins are replaced by corresponding non-human residues to improve antigen binding affinity.

[0066] In alternative embodiments, humanized antibodies may include residues not found in the recipient antibody or the donor antibody. These modifications may be performed to improve antibody affinity or functional activity. In alternative embodiments, humanized antibodies may include substantially all of at least one, usually two variable domains, wherein all or substantially all of the hypervariable regions correspond to the hypervariable regions of non-human immunoglobulins, and all or substantially all of the antibody framework regions are antibody framework regions of human immunoglobulin sequences.

[0067] In alternative embodiments, humanized antibodies used to practice the embodiments provided herein can include at least a portion of an immunoglobulin constant region (Fc), typically an Fc region of a human immunoglobulin or an Fc region derived from a human immunoglobulin.

[0068] However, in alternative embodiments, fully human antibodies can also be used to implement the embodiments provided herein, and fully human antibodies include human antibodies, and the human antibodies include an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a person. This definition of human antibodies specifically excludes humanized antibodies including non-human antigen-binding residues.

[0069] In alternative embodiments, the method comprises using a humanized antibody that is capable of specifically binding to the αvβ3 (avb3) integrin polypeptide, including humanized VITAXIN TM (MedImmune) or MEDI-523, edalizumab, or MEDI-522 or ABEGRIN TM (MedImmune).

[0070] In an alternative embodiment, the method comprises using a humanized antibody, such as VITAXIN TM , as described in U.S. Pat. No. 6,590,079, U.S. Pat. No. 7,422,744 and U.S. Pat. No. 7,422,745. In alternative embodiments, antibodies for implementing the embodiments provided herein include antibodies to α v β 3 An antibody exhibiting selective binding affinity and comprising a heavy chain polypeptide and a light chain polypeptide, the heavy chain polypeptide comprising Figure 1A The variable region amino acid sequence (SEQ ID NO: 1) or a functional region fragment thereof, the light chain polypeptide comprises Figure 1B The variable region amino acid sequence shown (SEQ ID NO: 2) or a functional region fragment thereof.

[0071] In alternative embodiments, the antibody for implementing the embodiments provided herein includes " affinity maturation " antibody, for example, in one or more hypervariable regions, include one or more changed antibodies, compared with the parental antibody without these changes, these changes cause the antibody to improve the affinity of antigen (for example histone methyl and / or acetyltransferase). In alternative embodiments, the antibody for implementing the embodiments provided herein is a mature antibody, which has nanomolar or even picomolar affinity to the target antigen (for example, histone methyl and / or acetyltransferase). The antibody of affinity maturation can be produced by methods known in the art.

[0072] In alternative embodiments, the antibody used to implement the methods provided herein is: monoclonal antibody LM609; monoclonal antibody LM609 is described in the following literature, for example, in Cheresh et al., Journal of Biological Chemistry (J Biol Chem.), 1987, Vol. 262, No. 36, pp. 17703-17711, and U.S. Pat. No. 5,753,230, and U.S. Pat. No. 6,590,079. LM609 is a mouse monoclonal antibody specific for integrin αvβ3, see, for example, Cheresh, DA, Proc. Natl. Acad. Sci. USA, Vol. 84, pp. 6471-6475 (1987), and Cheresh et al., Journal of Biological Chemistry (J. Biol. Chem.), Vol. 262, pp. 17703-17711 (1987). LM609 was generated against and reacts with the M21 cell adhesion receptor (now called integrin αvβ3).

[0073] LM609 inhibits M21 cell ligation to αvβ3 ligands such as vitronectin, fibrinogen, and von Willebrand factor (Cheresh and Spiro, supra), and is also an inhibitor of αvβ3-mediated pathologies such as tumor-induced angiogenesis (Brooks et al., Cell, Vol. 79, pp. 1157-1164 (1994)), granulation tissue development in skin wounds (Clark et al., Am. J. Pat., 2003, pp. 1157-1164 (1994)), and αvβ3-mediated inflammatory responses (Brooks et al., Cell, Vol. 79, pp. 1157-1164 (1994)). 148, pp. 1407-1421 (1996)) and smooth muscle cell migration, such as occurs during restenosis (Choi et al., J. Vascular Surg., Vol. 19, pp. 125-134 (1994); Jones et al., Proc. Natl. Acad. Sci., Vol. 93, pp. 2482-2487 (1996)).

[0074] In alternative embodiments, antibodies useful in practicing the methods provided herein include antibodies to α v β 3 A grafted LM609-grafted antibody exhibiting selective binding affinity, comprising a heavy chain polypeptide and a light chain polypeptide, the heavy chain polypeptide comprising Figure 1A The variable region amino acid sequence (SEQ ID NO: 2) or a functional fragment thereof, the light chain polypeptide basically comprises the variable region amino acid sequence (SEQ ID NO: 9) and the variable region amino acid sequence (SEQ ID NO: 10) respectively (see Figure 3 ) or a functional fragment thereof.

[0075] Manufacturing products and kits

[0076] Provided are articles of manufacture and kits for practicing the methods provided herein, comprising a human antibody or a humanized antibody capable of specifically binding to an αvβ3 (avb3) polypeptide expressed on the surface of a cell, the Fc region of which has a selective affinity for human FcγRI (CD64) but has no or substantially no selective affinity for other FcγRs; and / or also comprising macrophages, neutrophils and dendritic cells; and optionally also comprising instructions for practicing the methods provided herein.

[0077] Any of the above aspects and embodiments may be combined with any other aspect or embodiment disclosed herein as in the "Summary" and / or "Detailed Description" sections.

[0078] As used in this specification and the claims, the singular form "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0079] Unless otherwise stated or obvious from the context, as used herein, the term "or" should be construed as inclusive and encompasses both "or" and "and."

[0080] Unless otherwise specified or obvious from the context, as used herein, the term "about" should be understood as being within the normal tolerance range of the field, such as within 2 standard deviations of the mean value. About (the use of the term "about") can be understood as being 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 value. Unless the context clearly indicates otherwise, all numerical values ​​provided herein are modified by the term "about".

[0081] Unless otherwise specified or obvious from the context, as used herein, the terms "substantially all," "substantially a majority," "substantially all," or "majority" encompass at least about 90%, 95%, 97%, 98%, 99%, or 99.5% or more of a reference amount of a composition.

[0082] The entire contents of each patent, patent application, publication and document cited herein are incorporated herein by reference. The citation of the above patents, patent applications, publications and documents does not admit that any of the above contents are relevant prior art, nor does it constitute any admission of the contents or dates of these publications or documents. The separate citation of these documents should not be interpreted as an assertion or admission that any part of the contents of any document is considered to be necessary material to meet the statutory disclosure requirements of patent applications in any country or region. Nevertheless, the right to rely on any such document to provide materials deemed by the examining body or court to be essential to the subject matter claimed is reserved where appropriate.

[0083] Without departing from the basic aspects of the present invention, the foregoing may be modified. Although the present invention has been described in full detail with reference to one or more specific embodiments, it will be appreciated by those of ordinary skill in the art that the embodiments specifically disclosed in this application may be changed, and these modifications and improvements are still within the scope and spirit of the present invention. In the absence of any element not specifically disclosed herein, the present invention exemplarily described herein may be appropriately implemented. Therefore, for example, in each case herein, any one of the terms "comprising", "consisting essentially of" and "consisting of" may be replaced by any one of the other two terms. Therefore, the terms and expressions employed are used as descriptive rather than restrictive terms, do not exclude equivalents or portions thereof of the features shown and described, and recognize that various modifications are possible within the scope of the present invention.

[0084] Several embodiments of the present invention have been described. However, it will be appreciated that various modifications may be made without departing from the spirit and scope of the present invention. Accordingly, other embodiments are within the scope of the following claims.

Claims

1. Use of a human antibody or a humanized antibody in the preparation of a medicament for treating or ameliorating cancer, wherein the αvβ3 polypeptide is expressed on the cell surface of the cancer, wherein the human antibody or humanized antibody can specifically bind to the human FcγRI (CD64) receptor expressed on human macrophages but not to other human FcγRs in the Fc region, and can specifically bind to the αvβ3 (avb3) polypeptide expressed on the cell surface, Thereby inducing an antibody-dependent cell-mediated cytotoxicity (ADCC) response against the cancer.

2. The use according to claim 1, wherein the cancer is a drug-resistant cancer.

3. The use according to claim 1 or 2, wherein the medicament is formulated for administration together with another cancer therapeutic agent or therapy, wherein optionally, the other cancer therapeutic agent comprises paclitaxel.

4. The use according to claim 1 or 2, wherein the medicament is formulated for administration at a dose comprising 1 mg / kg to 8 mg / kg or 0.5 mg / kg to 12 mg / kg of the human antibody or humanized antibody.

5. The method according to claim 1 or 2, wherein the drug is administered intravenously (IV), intrathecally, sublingually, rectally, intravaginally, subcutaneously, orally or intramuscularly (IM), or injected or placed in situ near, adjacent to or inside the cancer, or administered by placing or inserting an implant comprising the human antibody or humanized antibody in situ.

6. The use of claim 3, wherein the additional cancer therapeutic agent or therapy comprises an antibody selected from the group consisting of abavotumab, adelimumab, afutuzumab, alemtuzumab, atumomab, amatumomab, maanatumomab, acitumomab, baviximab, betumomab, bevacizumab, bivacizumab, blinatumomab, brentuximab, cantuzumab, catumaxomab, cetuximab, situximab, citumomab , clituzumab, canagliflozin, daratumumab, trazitumab, dulitumab, dustuzumab, demostatumab, dasitumumab, dalozumab, emeticizumab, elotuzumab, entuximab, ertuinomab, fatuinomab, felatuzumab, fentolimumab, falotuzumab, vortuximab, ganituzumab, gemtuzumab, gemtuximab, gebatumumab, ibritumomab tiuxetan, igovotumab, imaclizumab, intuximab, ointuzumab, intuximab monoclonal antibody, ipilimumab, itumumab, labetuzumab, lexatumumab, lintuzumab, lovotuzumab, rucamumab, mapatumumab, matuzumab, miratumumab, mitumumab, mitumumab, moxitumomab, nanatuzumab, naptuzumab, nexitumomab, nimotuzumab, novatumumab, okatuzumab, ofatumumab, olaratumab, onatuzumab, mooxetuzumab, ogavuzumab, panitumumab, pasatuzumab, paltratumumab , pemuzumab, pertuzumab, pintumomab, privitatumomab, rantumomab, rantrastuzumab, rituximab, rituximab, rotuximab, satumomab, sirolimus, silotuximab, cintuzumab, solitotumab, talizumab, talizumab, talitumomab, tetuzumab, tetuzumab, tetuzumab, tegazumab, tositumomab, trastuzumab, tukavicuzumab, utoximab, veltuzumab, worse thanumomab, vortumomab, zalutumumab and / or any combination thereof.

7. The use according to claim 3, wherein the additional cancer therapeutic agent or therapy comprises a growth factor inhibitor, wherein optionally the growth factor inhibitor comprises a receptor tyrosine kinase (RTK) inhibitor, a Src inhibitor, an antimetabolite inhibitor, gemcitabine, GEMZAR TM , mitotic poisons, paclitaxel, taxol, ABRAXANE TM , erlotinib, TARCEVA TM , Lapatinib, TYKERB TM , Cetuximab, ERBITUX TM , PD-1 inhibitors, PD-L1 inhibitors and / or insulin growth factor inhibitors.

8. The use according to claim 1 or 2, wherein before administration to an individual in need thereof, a plurality of the drugs are pre-incubated in vitro with the human macrophages, neutrophils, monocytes and / or dendritic cells, wherein optionally the human macrophages, neutrophils, monocytes and / or dendritic cells are activated human macrophages, neutrophils, monocytes and / or dendritic cells.

9. Use of a human antibody or a humanized antibody in the preparation of a drug for treating or ameliorating cancer, wherein the human antibody or the humanized antibody is capable of specifically binding to the human FcγRI (CD64) receptor expressed on the surface of human macrophages in the Fc region but not binding to other human FcγRs, and is capable of specifically binding to the αvβ3 (avb3) polypeptide expressed on the cell surface, wherein the αvβ3 polypeptide is expressed on the cell surface of the cancer.

10. A human antibody or a humanized antibody, wherein the Fc region of the human antibody or the humanized antibody is capable of specifically binding to the human FcγRI (CD64) receptor expressed on the surface of human macrophages but not to other human FcγRs, and is capable of specifically binding to the αvβ3 (avb3) polypeptide expressed on the cell surface, wherein the human antibody or the humanized antibody is used to treat or improve cancer, wherein the αvβ3 polypeptide is expressed on the cell surface of the cancer.

Images