# Patent

## Title
Therapeutic combination of a VEGF antagonist and anti-hypertensive agent

## Abstract

Disclosed are compositions and methods for treating a disease or condition related to angiogenesis with a vascular endothelial growth factor (VEGF) inhibitor and one or more anti-hypertensive agent(s). The method of the invention is useful for preventing the development of hypertension and/or reducing hypertension in a subject treated with a VEGF inhibitor.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit under 35 USC § 119(e) of U.S. Provisional 60/652,394 filed 11 Feb. 2005, which application is herein specifically incorporated by reference in its entirety.


BACKGROUND
1. Field of the Invention
The field of the invention is related to therapeutic methods of treating diseases in a mammal with a vascular endothelial growth factor (VEGF) antagonist in combination with one or more anti-hypertensive agents.
2. Description of Related Art
Vascular endothelial growth factor (VEGF) has been recognized as a primary stimulus of angiogenesis in pathological conditions. Approaches to methods of blocking VEGF include soluble receptor constructs, antisense molecules, RNA aptamers, and antibodies. See, for example, PCT WO/0075319, for a description of VEGF-receptor based antagonists.
Combination therapies using an anti-VEGF antibody and chemotherapeutic agents, such as paclitaxel (TAXOLTM), are known (see, for example, U.S. Pat. No. 6,342,219).
BRIEF SUMMARY OF THE INVENTION
In one aspect, the invention features a pharmaceutical composition comprising a high affinity vascular endothelial cell growth factor (VEGF) antagonist, one or more anti-hypertensive therapeutic agent(s), and a pharmaceutically acceptable carrier.
More specifically, the VEGF antagonist a high affinity fusion protein dimer (or “trap”) comprising a fusion polypeptide having an immunoglobulin-like (Ig) domain 2 of the VEGF receptor Flt1 and Ig domain 3 of the VEGF receptor Flk1 or Flt4, and a multimerizing component. Even more specifically, the VEGF antagonist comprises a fusion polypeptide selected from the group consisting of Flt1D2.Flk1D3.FcΔC1(a) (SEQ ID NOs:1-2), VEGFR1R2-FcΔC1 (a) (SEQ ID NOs:3-4), or a functionally equivalent thereof.
In specific embodiments, the one or more anti-hypertensive therapeutic agent are selected from the group consisting of ACE inhibitors (ACCUPRILTM (Parke-Davis); ALTACETM (Monarch); CAPTOPRILTM (Mylan); ENALAPRILATETM (Baxter); LOTENSINTM (Novartis); MAVIKTM (Bristol-Myers Squibb); PRINIVILTM (Merck); UNIVASCTM (Schwarz), VASOTECTM (Merck); calcium-channel antagonists such as nifedipine, β-adrenergic receptor antagonists, such as for example, propanalol, sotalol; angiotensin II receptor antagonists; α-adrenergic receptor antagonists; direct active vasodilators; and diuretic agents used in the treatment of hypertension. In preferred embodiments, the anti-hypertensive therapeutic agent is an ACE inhibitor or a β-adrenergic receptor blocker.
In a second aspect, the invention features a pharmaceutical composition comprising a vascular endothelial cell growth factor (VEGF) antagonist, one or more anti-hypertensive therapeutic agent(s), and a pharmaceutically acceptable carrier, wherein the VEGF antagonist is a dimer composed of two fusion proteins each having an immunoglobulin-like (Ig) domain 2 of the VEGF receptor Flt1 and Ig domain 3 of the VEGF receptor Flk1 or Flt4, and a multimerizing component. In specific embodiments, the VEGF antagonist is selected from the group consisting of Flt1D2.Flk1D3.FcΔC1(a) (SEQ ID NOs:1-2), VEGFR1R2-FcΔC1(a) (SEQ ID NOs:3-4), or a functionally equivalent thereof.
In a third aspect, the invention features a method of treating a disease or condition which is ameliorated, inhibited, or reduced by a VEGF antagonist in a human, comprising administering a combination of a vascular endothelial growth factor (VEGF) antagonist and at least one anti-hypertensive agent.
The combined therapeutics of the invention achieves maximal anti-angiogenic activity while minimizing the known side effects resulting from treatment with anti-angiogenic agents, specifically, hypertension. The combination of an anti-angiogenic agent with an ACE inhibitor or angiotensin receptor blocker may also be used to prevent proteinuria in subjects at risk thereof.
Diseases and/or conditions, or recurrences thereof, which are ameliorated, inhibited, or reduced by treatment with the combination of the invention are those treated with a VEGF inhibitor such as the VEGF trap described above. For example, conditions ameliorated by treatment with a VEGF inhibitor include diseases such as cancer or diabetes. Conditions which are ameliorated, inhibited, prevented, or reduced by treatment with the combined therapeutics of the invention include vascular permeability, edema, or inflammation such as brain edema associated with injury, stroke, or tumor, edema associated with inflammatory disorders such as psoriasis or arthritis, asthma, edema associated with burns, ascites and pleural effusion associated with tumors, inflammation or trauma, chronic airway inflammation, capillary leak syndrome, sepsis kidney disease associated with increased leakage of protein, eye disorders such as eye related macular degeneration and diabetic retinopathy, abnormal angiogenesis such as polycystic ovary disease, entometriosis and endometrial carcinoma. A VEGF inhibitor may also be used to induce regression or reduction of the size of an existing tumor or metastatic cancer; diabetes, decrease tumor neovascularization, improve transplant corneal survival time, inhibit corneal transplant rejection or corneal lympangiogenesis and angiogenesis.
A subject to be treated is preferably a subject with one of the above listed conditions who suffers from hypertension, is at risk for development of hypertension or in which the prevention or inhibition of hypertension is desirable, e.g., a subject at risk for cardiovascular disease, a subject over 65 years of age, or a patient who cannot otherwise be treated with an appropriate dose of the VEGF antagonist without developing hypertension.
The VEGF inhibitor and anti-hypertensive agent may be administered simultaneously, separately or in combination, or sequentially over a relatively short period of time, e.g., within minutes, hours, or days.
In a fourth aspect, the invention features a method of preventing the development of hypertension during treatment with a vascular endothelial growth factor (VEGF) inhibitor in a patient at risk thereof, comprising administering a combination of a VEGF) antagonist and at least one anti-hypertensive agent.
In a fifth aspect, the invention features a method of treating hypertension during treatment with a vascular endothelial growth factor (VEGF) inhibitor in a patient at risk thereof, comprising administering a combination of a VEGF) antagonist and at least one anti-hypertensive agent.
Other objects and advantages will become apparent from a review of the ensuing detailed description.


DETAILED DESCRIPTION
Before the present methods and compositions are described, it is to be understood that this invention is not limited to particular methods, and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.
As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Thus for example, a reference to “a method” includes one or more methods, and/or steps of the type described herein and/or which will become apparent to those persons skilled in the art upon reading this disclosure and so forth.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to describe the methods and/or materials in connection with which the publications are cited.
General Description
In the normal mammal, blood pressure is strictly controlled by a complex system of physiological factors. This is important for survival because high blood pressure (hypertension) can lead to a number of adverse medical events and conditions, such as, for example, stroke, acute coronary syndrome, myocardial infarction, and renal failure. Studies show that VEGF transiently dilates coronary arteries in vitro (Ku et. al. (1993) Am J Physiol 265:H585-H592) and to induce hypotension (Yang et. al. (1996) J Cardiovasc Pharmacol 27:838-844). Methods for treating eclampsia and preemclampsia are known (see, for example, US patent application publication 2003/0220262, WO 98/28006, WO 00/13703) is described a method for treating hypertension comprising administering to a patient an effective amount of an angiogenic factor such as VEGF, or an agonist thereof. US patent application publication 2003/0144298 shows that administration of high levels of a VEGF receptor tyrosine kinase inhibitor leads to a sustained increase in blood pressure in rats when administered chronically.
VEGF Antagonists and VEGF-Specific Fusion Polypeptide Traps
In a preferred embodiment, the VEGF antagonist is a dimeric fusion protein capable of binding VEGF with a high affinity composed of two receptor-Fc fusion protein consisting of the principal ligand-binding portions of the human VEGFR1 and VEGFR2 receptor extracellular domains fused to the Fc portion of human IgG1 (termed a “VEGF trap”). Specifically, the VEGF trap consists of Ig domain 2 from VEGFR1, which is fused to Ig domain 3 from VEGFR2, which in turn is fused to the Fc domain of IgG1 (SEQ ID NO:2).
In a preferred embodiment, an expression plasmid encoding the VEGF trap is transfected into CHO cells, which secrete VEGF trap into the culture medium. The resulting VEGF trap is a dimeric glycoprotein with a protein molecular weight of 97 kDa and contains  ̃15% glycosylation to give a total molecular weight of 115 kDa.
Since the VEGF trap binds its ligands using the binding domains of high-affinity receptors, it has a greater affinity for VEGF than do monoclonal antibodies. The VEGF trap binds VEGF-A (KD=1.5 pM), PLGF1 (KD=1.3 nM), and PLGF2 (KD=50 pM); binding to other VEGF family members has not yet been fully characterized.
Anti-Hypertensive Therapeutic Agents
The invention may be practiced with a VEGF inhibitor, preferably a VEGF trap as described in U.S. Pat. No. 6,833,349, herein specifically incorporated by reference, and an agent which is capable of lowering blood pressure. Anti-hypertensive agents include calcium channel blockers, angiotensin converting enzyme inhibitors (ACE inhibitors), angiotensin II receptor antagonists (A-II antagonists), diuretics, β-adrenergic receptor blockers, vasodilators and α-adrenergic receptor blockers.
Calcium channel blockers include amlodipine; bepridil; clentiazem; diltiazem; fendiline; gallopamil; mibefradil; prenylamine; semotiadil; terodiline; verapamil; aranidipine; barnidipine; benidipine; cilnidipine; efonidipine; elgodipine; felodipine; isradipine; lacidipine; lercanidipine; manidipine; nicardipine; nifedipine; nilvadipine; nimodipine; nisoldipine; nitrendipine; cinnarizine; flunarizine; lidoflazine; lomerizine; bencyclane; etafenone; and perhexiline.
Angiotensin converting enzyme inhibitors (ACE-inhibitors) include alacepril; benazepril; captopril; ceronapril; delapril; enalapril; fosinopril; imidapril; lisinopril; moveltipril; perindopril; quinapril; ramipril; spirapril; temocapril; and trandolapril.
Angiotensin-II receptor antagonists include, but are not limited to: candesartan (U.S. Pat. No. 5,196,444); eprosartan; irbesartan; losartan; and valsartan.
β-blockers include, but are not limited to: acebutolol; alprenolol; amosulalol; arotinolol; atenolol; befunolol; betaxolol; bevantolot; bisoprolol; bopindolol; bucumolol; bufetolol; bufuralol; bunitrolol; bupranolol; butidrine hydrochloride; butofilolol; carazolol; carteolol; carvedilol; celiprolol; cetamolol; cloranololdilevalol; epanolol; indenolol; labetalol; levobunolol; mepindolol; metipranolol; metoprolol; moprolol; nadolol; nadoxolol; nebivalol; nipradilol; oxprenolol; penbutolol; pindolol; practolol; pronethalol; propranolol; sotalol; sulfinalol; talinolol; tertatolol; tilisolol; timolol; toliprolol; and xibenolol.
α-blockers include, but are not limited to: amosulalol; arotinolol; dapiprazole; doxazosin; fenspiride; indoramin; labetolol, naftopidil; nicergoline; prazosin; tamsulosin; tolazoline; trimazosin; and yohimbine.
Vasodilators include cerebral vasodilators, coronary vasodilators and peripheral vasodilators. Cerebral vasodilators include bencyclane; cinnarizine; citicoline; cyclandelate; ciclonicate; diisopropylamine dichloroacetate; eburnamonine; fasudil; fenoxedil; flunarizine; ibudilast; ifenprodil; lomerizine; nafronyl; nicametate; nicergoline; nimodipine; papaverine; tinofedrine; vincamine; vinpocetine; and viquidil.
Coronary vasodilators include, but are not limited to: amotriphene; bendazol; benfurodil hemisuccinate; benziodarone; chloracizine; chromonar; clobenfural; clonitrate; cloricromen; dilazep; dipyridamole; droprenilamine; efloxate; erythrityl tetranitrate; etafenone; fendiline; floredil; ganglefene; hexestrol bis(β-diethylaminoethyl) ether; hexobendine; itramin tosylate; khellin; lidoflazine; mannitol hexanitrate; medibazine; nitroglycerin; pentaerythritol tetranitrate; pentrinitrol; perhexiline; pimefylline; prenylamine; propatyl nitrate; trapidil; tricromyl; trimetazidine; trolnitrate phosphate; visnadine.
Peripheral vasodilators include, but are not limited to: aluminium nicotinate; bamethan; bencyclane; betahistine; bradykinin; brovincamine; bufeniode; buflomedil; butalamine; cetiedil; ciclonicate; cinepazide; cinnarizine; cyclandelate; diisopropylamine dichloroacetate; eledoisin; fenoxedil; flunarizine; hepronicate; ifenprodil; iloprost; inositol niacinate; isoxsuprine; kallidin; kallikrein; moxisylyte; nafronyl; nicametate; nicergoline; nicofuranose; nylidrin; pentifylline; pentoxifylline; piribedil; prostaglandin E1; suloctidil; tolazoline; and xanthinol niacinate.
Diuretic includes but is not limited to diuretic benzothiadiazine derivatives, diuretic organomercurials, diuretic purines, diuretic steroids, diuretic sulfonamide derivatives, diuretic uracils and other diuretics such as amanozine; amiloride; arbutin; chlorazanil; ethacrynic acid; etozolin; hydracarbazine; isosorbide; mannitol; metochalcone; muzolimine; perhexiline; ticrynafen; triamterene; and urea.
Treatment Population
A human subject preferably treated with the combined therapeutics described herein is a subject in which it is desirable to prevent or reduce one or more side effects resulting from treatment with an anti-angiogenic agent, such as hypertension, proteinuria. Particularly preferred subjects are those suffering from hypertension, over 65 years of age, or subjects in which reduction of or prevention of undesirable side effects allows a maximal dose of the anti-angiogenic agent to be used which otherwise could not be used without placing the subject at risk for an adverse medical event. Patients suffering from renal cell carcinoma, pancreatic carcinoma, advanced breast cancer, colorectal cancer, malignant mesothelioma, multiple myeloma, ovarian cancer, or melanoma may be treated with the combined therapeutics of the invention. Diseases and/or conditions, or recurrences thereof, which are ameliorated, inhibited, or reduced by treatment with the combined therapeutics of the invention cancer, diabetes, vascular permeability, edema, or inflammation such as brain edema associated with injury, stroke, or tumor, edema associated with inflammatory disorders such as psoriasis or arthritis, asthma, edema associated with burns, ascites and pleural effusion associated with tumors, inflammation or trauma, chronic airway inflammation, capillary leak syndrome, sepsis kidney disease associated with increased leakage of protein, eye disorders such as eye related macular degeneration and diabetic retinopathy, abnormal angiogenesis such as polycystic ovary disease, entometriosis and endometrial carcinoma. A VEGF inhibitor may also be used to induce regression or reduction of the size of an existing tumor or metastatic cancer; diabetes, decrease tumor neovascularization, improve transplant corneal survival time, inhibit corneal transplant rejection or corneal lympangiogenesis and angiogenesis.
Combination Therapies
In numerous embodiments, a VEGF antagonist may be administered in combination with one or more additional compounds or therapies, including a second VEGF antagonist molecule. Combination therapy includes administration of a single pharmaceutical dosage formulation which contains a VEGF antagonist and one or more additional agents; as well as administration of a VEGF antagonist and one or more additional agent(s) in its own separate pharmaceutical dosage formulation. For example, a VEGF antagonist and a cytotoxic agent, a chemotherapeutic agent or a growth inhibitory agent can be administered to the patient together in a single dosage composition such as a combined formulation, or each agent can be administered in a separate dosage formulation. Where separate dosage formulations are used, the VEGF-specific fusion protein of the invention and one or more additional agents can be administered concurrently, or at separately staggered times, i.e., sequentially.
The term “cytotoxic agent” as used herein refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes (e.g. I131, I125, Y90 and Re186), chemotherapeutic agents, and toxins such as enzymatically active toxins of bacterial, fungal, plant or animal origin, or fragments thereof.
A “chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2′′-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g. paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) and docetaxel (TAXOTERE®; Aventis Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine; retinoic acid; esperamicins; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above.
A “growth inhibitory agent” when used herein refers to a compound or composition which inhibits growth of a cell, especially a cancer cell either in vitro or in vivo. Examples of growth inhibitory agents include agents that block cell cycle progression (at a place other than S phase), such as agents that induce G1 arrest and M-phase arrest. Classical M-phase blockers include the vincas (vincristine and vinblastine), TAXOL®, and topo II inhibitors such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin. Those agents that arrest G1 also spill over into S-phase arrest, for example, DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C.
Methods of Administration
The invention provides compositions and methods of treatment comprising a VEGF antagonist, such as a VEGF antagonist, and an anti-hypertensive agent. Various delivery systems are known and can be used to administer the composition of the invention, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the compound, receptor-mediated endocytosis (see, e.g., Wu and Wu, 1987, J. Biol. Chem. 262:4429-4432), construction of a nucleic acid as part of a retroviral or other vector, etc. Methods of introduction can be enteral or parenteral and include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, intraocular, and oral routes. The compounds may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local. Administration can be acute or chronic (e.g. daily, weekly, monthly, etc.) or in combination with other agents. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.
In another embodiment, the active agent can be delivered in a vesicle, in particular a liposome, in a controlled release system, or in a pump. In another embodiment where the active agent of the invention is a nucleic acid encoding a protein, the nucleic acid can be administered in vivo to promote expression of its encoded protein, by constructing it as part of an appropriate nucleic acid expression vector and administering it so that it becomes intracellular, e.g., by use of a retroviral vector (see, for example, U.S. Pat. No. 4,980,286), by direct injection, or by use of microparticle bombardment, or coating with lipids or cell-surface receptors or transfecting agents, or by administering it in linkage to a homeobox-like peptide which is known to enter the nucleus.
In a specific embodiment, it may be desirable to administer the pharmaceutical compositions of the invention locally to the area in need of treatment; this may be achieved, for example, and not by way of limitation, by local infusion during surgery, topical application, e.g., by injection, by means of a catheter, or by means of an implant, the implant being of a porous, non-porous, or gelatinous material, including membranes, such as silastic membranes, fibers, or commercial skin substitutes.
A composition useful in practicing the methods of the invention may be a liquid comprising an agent of the invention in solution, in suspension, or both. The term “solution/suspension” refers to a liquid composition where a first portion of the active agent is present in solution and a second portion of the active agent is present in particulate form, in suspension in a liquid matrix. A liquid composition also includes a gel. The liquid composition may be aqueous or in the form of an ointment.
An aqueous suspension or solution/suspension useful for practicing the methods of the invention may contain one or more polymers as suspending agents. Useful polymers include water-soluble polymers such as cellulosic polymers and water-insoluble polymers such as cross-linked carboxyl-containing polymers. An aqueous suspension or solution/suspension of the present invention is preferably viscous or muco-adhesive, or even more preferably, both viscous and mucoadhesive.
Pharmaceutical Compositions
The present invention provides pharmaceutical compositions comprising a VEGF antagonist, an anti-hypertensive agent, and a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly, in humans. The term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E. W. Martin.
The composition of the invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.
The amount of the composition of the invention that will be effective for its intended therapeutic use can be determined by standard clinical techniques based on the present description. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. Generally, suitable dosage ranges for intravenous administration are generally about 20-500 micrograms of active compound per kilogram body weight. Suitable dosage ranges for intranasal administration are generally about 0.01 pg/kg body weight to 1 mg/kg body weight. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.
For systemic administration, a therapeutically effective dose can be estimated initially from in vitro assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC50 as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Initial dosages can also be estimated from in vivo data, e.g., animal models, using techniques that are well known in the art. One having ordinary skill in the art could readily optimize administration to humans based on animal data.
Dosage amount and interval may be adjusted individually to provide plasma levels of the compounds that are sufficient to maintain therapeutic effect. In cases of local administration or selective uptake, the effective local concentration of the compounds may not be related to plasma concentration. One having skill in the art will be able to optimize therapeutically effective local dosages without undue experimentation.
The amount of compound administered will, of course, be dependent on the subject being treated, on the subject's weight, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician. The therapy may be repeated intermittently while symptoms are detectable or even when they are not detectable. The therapy may be provided alone or in combination with other drugs.
Kits
The invention also provides an article of manufacturing comprising packaging material and a pharmaceutical agent contained within the packaging material, wherein the pharmaceutical agent comprises at least one VEGF antagonist and at least one anti-hypertensive agent, and wherein the packaging material comprises a label or package insert which indicates that the VEGF antagonist and anti-hypertensive agent can be used for treating cancer or reducing tumor growth.
Other features of the invention will become apparent in the course of the following descriptions of exemplary embodiments which are given for illustration of the invention and are not intended to be limiting thereof.
EXAMPLES
The following example is put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.
Example 1
Treatment of Malignant Pleural Effusion and Prevention of Hypertension
Adult patients with pathologic diagnosis of stage IIIB-IV NSCLC who are eligible for systemic chemotherapy, and also have an MPE which requires therapeutic drainage are eligible for inclusion in the study. Patients undergoing chemotherapy with another agent, or having prior chemotherapy with an inhibitor of VEGF, or active or untreated brain metastases are excluded. Treatment with VEGF antagonist (SEQ ID NO:4) is an intravenous dose of 300-5000 mg/kg and an anti-hypertensive agent such as an ACE inhibitor or a β-adrenergic receptor blocker. The anti-hypertensive therapeutic agent may be given separately or in combination with the VEGF antagonist, prior to administration of the VEGF antagonist, simultaneously, or following administration of the VEGF antagonist.
Example 2
Treatment of Solid Tumor and Prevention of Hypertension
Patients with refractory solid tumors or non-Hodgkin's lymphoma receiving no concurrent treatment for their cancer are treated with the VEGF trap (SEQ ID NO:4) as follows. The dose levels range from 100 to 5000 mg/kg given subcutaneously. Each patient receives a single initial dose of the VEGF trap followed by weekly injections at the required dose level. Blood pressure is monitored and tumor burden is assessed at the beginning and end of the weekly dosing period; patients with stable disease, partial or complete responses may continue dosing for up to an additional 6 months in a continuation study.

## Claims

1. A pharmaceutical composition comprising a high affinity vascular endothelial cell growth factor (VEGF) antagonist, one or more anti-hypertensive therapeutic agent(s), and a pharmaceutically acceptable carrier, wherein the VEGF antagonist is VEGFR1R2-FcΔC1(a) (SEQ ID NO:4).

2. The pharmaceutical composition of claim 1, wherein the one or more anti-hypertensive therapeutic agent is(are) selected from the group consisting of an angiotensin converting enzyme (ACE) inhibitor, a calcium-channel antagonist, a β-adrenergic receptor antagonist, an angiotensin II receptor antagonist, an α-adrenergic receptor antagonist, a vasodilator and a diuretic agent.

3. The pharmaceutical composition of claim 2, wherein the ACE inhibitor is selected from the group consisting of alacepril; benazepril; captopril; ceronapril; delapril; enalapril; fosinopril; imidapril; lisinopril; moveltipril; perindopril; quinapril; ramipril; spirapril; temocapril; and trandolapril.

4. The pharmaceutical composition of claim 2, wherein the calcium-channel blocker is selected from the group consisting of amlodipine, bepridil, clentiazem, diltiazem, fendiline, gallopamil, mibefradil, prenylamine, semotiadil, terodiline, verapamil, aranidipine, barnidipine, benidipine, cilnidipine, efonidipine, elgodipine, felodipine, isradipine, lacidipine, lercanidipine, manidipine, nicardipine, nifedipine, nilvadipine, nimodipine, nisoldipine, nitrendipine, cinnarizine, flunarizine, lidoflazine, lomerizine, bencyclane, etafenone, and perhexiline.

5. The pharmaceutical composition of claim 2, wherein the β-adrenergic receptor antagonist is selected from the group consisting of acebutolol, alprenolol, amosulalol, arotinolol, atenolol, befunolol, betaxolol, bevantolot, bisoprolol, bopindolol, bucumolol, bufetolol, bufuralol, bunitrolol, bupranolol, butidrine hydrochloride, butofilolol, carazolol, carteolol, carvedilol, celiprolol, cetamolol, cloranololdilevalol, epanolol, indenolol, labetalol, levobunolol, mepindolol, metipranolol, metoprolol, moprolol, nadolol, nadoxolol, nebivalol, nipradilol, oxprenolol, penbutolol, pindolol, practolol, pronethalol, propranolol, sotalol, sulfinalol, talinolol, tertatolol, tilisolol, timolol, toliprolol, and xibenolol.

6. The pharmaceutical composition of claim 2, wherein the α-adrenergic receptor antagonist is selected from the group consisting of amosulalol, arotinolol, dapiprazole, doxazosin, fenspiride, indoramin, labetolol, naftopidil, nicergoline, prazosin, tamsulosin, tolazoline, trimazosin, and yohimbine.

7. The pharmaceutical composition of claim 2, wherein the vasodilator is a cerebral vasodilator, a coronary vasodilator, and/or a peripheral vasodilator.

8. The pharmaceutical composition of claim 7, wherein the cerebral vasodilator is selected from the group consisting of bencyclane, cinnarizine, citicoline, cyclandelate, ciclonicate, diisopropylamine dichloroacetate, eburnamonine, fasudil, fenoxedil, flunarizine, ibudilast, ifenprodil, lomerizine, nafronyl, nicametate, nicergoline, nimodipine, papaverine, tinofedrine, vincamine, vinpocetine, and viquidil.

9. The pharmaceutical composition of claim 7, wherein the coronary vasodilator is selected from the group consisting of amotriphene, bendazol, benfurodil hemisuccinate, benziodarone, chloracizine, chromonar, clobenfural, clonitrate, cloricromen, dilazep, dipyridamole, droprenilamine, efloxate, erythrityl tetranitrate, etafenone, fendiline, floredil, ganglefene, hexestrol bis(β-diethylaminoethyl) ether, hexobendine, itramin tosylate, khellin, lidoflazine, mannitol hexanitrate, medibazine, nitroglycerin, pentaerythritol tetranitrate, pentrinitrol, perhexiline, pimefylline, prenylamine, propatyl nitrate, trapidil, tricromyl, trimetazidine, trolnitrate phosphate, visnadine.

10. The pharmaceutical composition of claim 7, wherein the peripheral vasodilator is selected from the group consisting of aluminium nicotinate, bamethan, bencyclane, betahistine, bradykinin, brovincamine, bufeniode, buflomedil, butalamine, cetiedil, ciclonicate, cinepazide, cinnarizine, cyclandelate, diisopropylamine dichloroacetate, eledoisin, fenoxedil, flunarizine, hepronicate, ifenprodil, iloprost, inositol niacinate, isoxsuprine, kallidin, kallikrein, moxisylyte, nafronyl, nicametate, nicergoline, nicofuranose, nylidrin, pentifylline, pentoxifylline, piribedil, prostaglandin E1, suloctidil, tolazoline, and xanthinol niacinate.

11. The pharmaceutical composition of claim 2, wherein diuretic agent is selected from the group consisting of benzothiadiazine derivatives, diuretic organomercurials, diuretic purines, diuretic steroids, diuretic sulfonamide derivatives, diuretic uracils, amanozine, amiloride, arbutin, chlorazanil, ethacrynic acid, etozolin, hydracarbazine, isosorbide, mannitol, metochalcone, muzolimine, perhexiline, ticrynafen, triamterene, and urea.