Therapeutic combination and methods of treatment with a DLL4 antagonist and an anti-hypertensive agent

1. A method of ameliorating hypertension in a subject receiving treatment with a delta-like ligand 4 (DLL4) antagonist, the method comprising:

administering to the subject a therapeutically effective amount of one or more anti-hypertensive agents to control hypertension caused by the antagonist,

wherein the DLL4 antagonist comprises a monoclonal antibody that specifically binds human DLL4.

2. The method of claim 1 , wherein the monoclonal antibody comprises an intact monoclonal antibody.

3. The method of claim 1 , wherein the monoclonal antibody is encoded by the plasmid having ATCC deposit no. PTA-8425.

4. The method of claim 1 , wherein the monoclonal antibody competes for specific binding to human DLL4 with an antibody encoded by the plasmid deposited with ATCC, having deposit no. PTA-8425.

5. The method of claim 1 , wherein the monoclonal antibody comprises: (i) a heavy chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO:1); CDR2 (SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4); and CDR3 (SEQ ID NO:5), and (ii) a light chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO:7); CDR2 (SEQ ID NO:8); and CDR3 (SEQ ID NO:9).

6. The method of claim 5 , wherein the heavy chain variable region comprises CDR amino acid sequences CDR1 (SEQ ID NO:1), CDR2 (SEQ ID NO:3), and CDR3 (SEQ ID NO:5), and the light chain variable region comprises CDR amino acid sequences CDR1 (SEQ ID NO:7); CDR2 (SEQ ID NO:8); and CDR3 (SEQ ID NO:9).

7. The method of claim 1 , wherein the monoclonal antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6.

8. The method of claim 7 , wherein the monoclonal antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:10.

9. The method of claim 1 , wherein the monoclonal antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:10.

10. The method of claim 1 , wherein the monoclonal antibody is a humanized, chimeric, or human antibody.

11. The method of claim 1 , wherein the monoclonal antibody comprises an antibody fragment.

12. The method of claim 11 , wherein the monoclonal antibody is a multispecific antibody.

13. The method of claim 1 , wherein the monoclonal antibody is a fusion protein that comprises an immunoglobulin molecule that specifically binds human DLL4 through at least one antigen recognition site within the variable region of the immunoglobulin molecule.

14. The method of claim 1 , wherein the anti-hypertensive agent is selected from the group consisting of a diuretic, an adrenergic receptor antagonist, an adrenergic receptor agonist, a calcium channel blocker, an ACE inhibitor, an angiotensin II receptor antagonist, an aldosterone antagonist, a vasodilator, a renin inhibitor, and combinations thereof.

15. The method of claim 14 , wherein the anti-hypertensive agent is a diuretic selected from the group, consisting of: furosemide, bumetanide, ethacrynic acid, torsemide, epitizide, hydrochlorothiazide, hydroflumethiazide, chlorothiazide bendroflumethiazide, polythiazide, trichlormethiazide cyclopenthiazide, methyclothiazide, cyclothiazide, mebutizide, indapamide, chlortalidone, metolazone, quinethazone, clopamide, mefruside, clofenamide, meticrane, xipamide, clorexolone, fenquizone, amiloride, triamterene, eplerenone, benzamil, potassium canrenoate, canrenone, spironolactone mannitol, glucose, urea, conivaptan, relcovaptan, nelivaptan, lixivaptan, mozavaptan, satavaptan, tolvaptan, demeclocycline, mersalyl acid, meralluride, mercaptomerin, mercurophyllme, merethoxyllme procaine, calomel, caffeine, theobromine, paraxanthine, theophylline, acetazolamide, methazolamide, dorzolamide, sulfonamide, topiramate, amanozine, arbutin, chlorazanil etozolin, hydracarbazine, isosorbide, metochalcone, muzohmme, perhexiline, and ticrynafen.

16. The method of claim 14 , wherein the anti-hypertensive agent is an adrenergic receptor antagonist selected from the group consisting of: atenolol, metoprolol, nadolol oxprenolol, pindolol, propranolol, timolol, acebutolol, bisoprolol, esmolol, labetalol, carvedilol, bucindolol, nebivolol, amosulalol, arotinolol, befunolol, betaxolol, bevantolot, bopindolol, bucumolol, bufetolol, bufuralol, bumtrolol, bupranolol, butidrine hydrochloride, butofilolol, carazolol, carteolol, celiprolol, cetamolol, cloranololdilevalol, epanolol, indenolol, levobunolol, mepindolol, metipranolol, moprolol, nadoxolol, nipradilol, penbutolol, practolol, pronethalol, sotalol, sulfmalol, talinolol, tertatolol, tilisolol, toliprolol, xibenolol, phenoxybenzamine, prazosin, doxazosin, terazosin, trimazosin, phentolamine, amosulalol, arotinolol, dapiprazole, fenspinde, indoramin, labetalol, naftopidil, nicergolme, tamsulosin, tolazolme, moxonidine, reserpine, and yohimbine.

17. The method of claim 14 , wherein the anti-hypertensive agent is an adrenergic receptor agonist selected from the group consisting of: clonidine, methyldopa, guanfacine, methoxamine, methylnorepinephrine, oxymetazoline, phenylephrine, guanabenz, guanoxabenz, guanethidine, xylazine, and tizanidme.

18. The method of claim 14 , wherein the anti-hypertensive agent is a calcium channel blocker selected from the group consisting of: amlodipine, felodipine, nicardipine, nifedipine, nimodipine, isradipine, nitrendipine, aranidipine, barnidipine, benidipine, cilnidipine, efonidipine, elgodipine, lacidipine, lercanidipine, manidipine, nilvadipine, nisoldipine, diltiazem, verapamil, bepridil, clentiazem, fendiline, gallopamil, mibefradil, prenylamine, semotiadil, terodiline, cinnarizine, flunarizine, lidoflazine, lomerizine, bencyclane, etafenone, and perhexiline.

19. The method of claim 14 , wherein the anti-hypertensive agent is an ACE inhibitor.

20. The method of claim 19 , wherein the ACE inhibitor is selected from the group consisting of: captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lismopril, benazepril, fosinopril, ceronapril, casokinins, lactokinins, teprotide, alacepril, cilazapril, delapril, imidapnl, moexipnl, rentiapril, spirapril, temocapril, moveltipnl and trandolapril.

21. The method of claim 14 , wherein the anti-hypertensive agent is an angiotensin II receptor antagonist selected from the group consisting of: candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan.

22. The method of claim 14 , wherein the anti-hypertensive agent is an aldosterone antagonist selected from the group consisting of: eplerenone, canrenone, and spironolactone.

23. The method of claim 14 , wherein the anti-hypertensive agent is a vasodilator 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, viquidil, amotriphene, bendazol, benfurodil hemisuccinate, benziodarone, chloracizme, chromonar, clobenfural, clonitrate, cloricromen, dilazep, dipyridamole, droprenilamine, efloxate, erythrityl tetranitrate, etafenone, fendiline, floredil, ganglefene, hexestrol bis(P-diethylaminoethyl) ether, hexobendine, itramin tosylate, khellin, lidoflazine, mannitol hexanitrate, medibazine, nitroglycerin, pentaerythritol tetranitrate, pentrinitrol, perhexiline, pimefylline, prenylamine, propatyl nitrate, trapidil, tricromyl, trimetazidine, trolnitrate phosphate, sildenafil, tadalafil, vardenafil, sodium nitroprusside, isosorbide mononitrate, isosorbide dinitrate, pentaerythritol, tetranitrate, theobromine, visnadine, aluminum nicotinate, bamethan, bencyclane, betahistine, brandykinin, 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, pentifyllme, pentoxifylline, piribedil, prostaglandin E1, suloctidil, tolazoline, and xanthinol niacinate.

24. The method of claim 14 , wherein the anti-hypertensive agent is a renin inhibitor selected from the group consisting of: aliskiren and remikiren.

25. The method of claim 1 , wherein the monoclonal antibody is an immunoglobulin molecule that specifically binds human DLL4 through at least one antigen recognition site within the variable region of the immunoglobulin molecule.

26. A method of detecting and ameliorating hypertension in a subject receiving treatment with a delta-like ligand 4 (DLL4) antagonist, the method comprising:

(a) measuring the blood pressure of the subject after receiving treatment with the DLL4 antagonist; and

(b) administering to the subject a therapeutically effective amount of one or more anti-hypertensive agents to control hypertension caused by the antagonist,

wherein the DLL4 antagonist comprises a monoclonal antibody that specifically binds human DLL4.

27. The method of claim 26 , wherein the monoclonal antibody comprises an intact monoclonal antibody.

28. The method of claim 26 , wherein the monoclonal antibody is encoded by the plasmid having ATCC deposit no. PTA-8425.

29. The method of claim 26 , wherein the monoclonal antibody competes for specific binding to human DLL4 with an antibody encoded by the plasmid deposited with ATCC, having deposit no. PTA-8425.

30. The method of claim 26 , wherein the monoclonal antibody comprises: (i) a heavy chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO:1); CDR2 (SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4); and CDR3 (SEQ ID NO:5), and (ii) a light chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO:7); CDR2 (SEQ ID NO:8); and CDR3 (SEQ ID NO:9).

31. The method of claim 30 , wherein the heavy chain variable region comprises CDR amino acid sequences CDR1 (SEQ ID NO:1), CDR2 (SEQ ID NO:3), and CDR3 (SEQ ID NO:5), and the light chain variable region comprises CDR amino acid sequences CDR1 (SEQ ID NO:7); CDR2 (SEQ ID NO:8); and CDR3 (SEQ ID NO:9).

32. The method of claim 26 , wherein the monoclonal antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6.

33. The method of claim 32 , wherein the monoclonal antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:10.

34. The method of claim 26 , wherein the monoclonal antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:10.

35. The method of claim 26 , wherein the monoclonal antibody is a humanized, chimeric, or human antibody.

36. The method of claim 26 , wherein the monoclonal antibody comprises an antibody fragment.

37. The method of claim 36 , wherein the monoclonal antibody is a multispecific antibody.

38. The method of claim 26 , wherein the monoclonal antibody is a fusion protein that comprises an immunoglobulin molecule that specifically binds human DLL4 through at least one antigen recognition site within the variable region of the immunoglobulin molecule.

39. The method of claim 26 , wherein the anti-hypertensive agent is selected from the group consisting of a diuretic, an adrenergic receptor antagonist, an adrenergic receptor agonist, a calcium channel blocker, an ACE inhibitor, an angiotensin II receptor antagonist, an aldosterone antagonist, a vasodilator, a renin inhibitor, and combinations thereof.

40. The method of claim 39 , wherein the anti-hypertensive agent is a diuretic selected from the group, consisting of: furosemide, bumetanide, ethacrynic acid, torsemide, epitizide, hydrochlorothiazide, hydroflumethiazide, chlorothiazide bendroflumethiazide, polythiazide, trichlormethiazide cyclopenthiazide, methyclothiazide, cyclothiazide, mebutizide, indapamide, chlortalidone, metolazone, quinethazone, clopamide, mefruside, clofenamide, meticrane, xipamide, clorexolone, fenquizone, amiloride, triamterene, eplerenone, benzamil, potassium canrenoate, canrenone, spironolactone mannitol, glucose, urea, conivaptan, relcovaptan, nelivaptan, lixivaptan, mozavaptan, satavaptan, tolvaptan, demeclocycline, mersalyl acid, meralluride, mercaptomerin, mercurophyllme, merethoxyllme procaine, calomel, caffeine, theobromine, paraxanthine, theophylline, acetazolamide, methazolamide, dorzolamide, sulfonamide, topiramate, amanozine, arbutin, chlorazanil etozolin, hydracarbazine, isosorbide, metochalcone, muzohmme, perhexiline, and ticrynafen.

41. The method of claim 39 , wherein the anti-hypertensive agent is an adrenergic receptor antagonist selected from the group consisting of: atenolol, metoprolol, nadolol oxprenolol, pindolol, propranolol, timolol, acebutolol, bisoprolol, esmolol, labetalol, carvedilol, bucindolol, nebivolol, amosulalol, arotinolol, befunolol, betaxolol, bevantolot, bopindolol, bucumolol, bufetolol, bufuralol, bumtrolol, bupranolol, butidrine hydrochloride, butofilolol, carazolol, carteolol, celiprolol, cetamolol, cloranololdilevalol, epanolol, indenolol, levobunolol, mepindolol, metipranolol, moprolol, nadoxolol, nipradilol, penbutolol, practolol, pronethalol, sotalol, sulfmalol, talinolol, tertatolol, tilisolol, toliprolol, xibenolol, phenoxybenzamine, prazosin, doxazosin, terazosin, trimazosin, phentolamine, amosulalol, arotinolol, dapiprazole, fenspinde, indoramin, labetalol, naftopidil, nicergolme, tamsulosin, tolazolme, moxonidine, reserpine, and yohimbine.

42. The method of claim 39 , wherein the anti-hypertensive agent is an adrenergic receptor agonist selected from the group consisting of: clonidine, methyldopa, guanfacine, methoxamine, methylnorepinephrine, oxymetazoline, phenylephrine, guanabenz, guanoxabenz, guanethidine, xylazine, and tizanidme.

43. The method of claim 39 , wherein the anti-hypertensive agent is a calcium channel blocker selected from the group consisting of: amlodipine, felodipine, nicardipine, nifedipine, nimodipine, isradipine, nitrendipine, aranidipine, barnidipine, benidipine, cilnidipine, efonidipine, elgodipine, lacidipine, lercanidipine, manidipine, nilvadipine, nisoldipine, diltiazem, verapamil, bepridil, clentiazem, fendiline, gallopamil, mibefradil, prenylamine, semotiadil, terodiline, cinnarizine, flunarizine, lidoflazine, lomerizine, bencyclane, etafenone, and perhexiline.

44. The method of claim 39 , wherein the anti-hypertensive agent is an ACE inhibitor.

45. The method of claim 44 , wherein the ACE inhibitor is selected from the group consisting of: captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lismopril, benazepril, fosinopril, ceronapril, casokinins, lactokinins, teprotide, alacepril, cilazapril, delapril, imidapnl, moexipnl, rentiapril, spirapril, temocapril, moveltipnl and trandolapril.

46. The method of claim 39 , wherein the anti-hypertensive agent is an angiotensin II receptor antagonist selected from the group consisting of: candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan.

47. The method of claim 39 , wherein the anti-hypertensive agent is an aldosterone antagonist selected from the group consisting of: eplerenone, canrenone, and spironolactone.

48. The method of claim 39 , wherein the anti-hypertensive agent is a vasodilator 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, viquidil, amotriphene, bendazol, benfurodil hemisuccinate, benziodarone, chloracizme, chromonar, clobenfural, clonitrate, cloricromen, dilazep, dipyridamole, droprenilamine, efloxate, erythrityl tetranitrate, etafenone, fendiline, floredil, ganglefene, hexestrol bis(P-diethylaminoethyl) ether, hexobendine, itramin tosylate, khellin, lidoflazine, mannitol hexanitrate, medibazine, nitroglycerin, pentaerythritol tetranitrate, pentrinitrol, perhexiline, pimefylline, prenylamine, propatyl nitrate, trapidil, tricromyl, trimetazidine, trolnitrate phosphate, sildenafil, tadalafil, vardenafil, sodium nitroprusside, isosorbide mononitrate, isosorbide dinitrate, pentaerythritol, tetranitrate, theobromine, visnadine, aluminum nicotinate, bamethan, bencyclane, betahistine, brandykinin, 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, pentifyllme, pentoxifylline, piribedil, prostaglandin E1, suloctidil, tolazoline, and xanthinol niacinate.

49. The method of claim 39 , wherein the anti-hypertensive agent is a renin inhibitor selected from the group consisting of: aliskiren and remikiren.

50. The method of claim 26 , wherein the subject is a human.

51. The method of claim 26 , wherein the monoclonal antibody is an immunoglobulin molecule that specifically binds human DLL4 through at least one antigen recognition site within the variable region of the immunoglobulin molecule.