(12) Unlted States Patent
`(10) Patent No.:
`US 8,101,659 B2
`
`Ksander et al.
`(45) Date of Patent:
`Jan. 24, 2012
`
`USOO8101659B2
`
`(54) METHODS OF TREATMENT AND
`PHARMACEUTICAL COMPOSITION
`
`(75)
`
`Inventors: Gary M Ksander, Amherst, NH (US);
`Randy L Webb, Flemington, NJ (US)
`
`(73) Assignee: NOVaI‘tiS AG, Basel (CH)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 15403) by 0 days.
`
`(21) Appl. NO': 12/147’570
`.
`Flled:
`
`Jun. 27, 2008
`
`(22)
`
`(65)
`
`Pri0r Publication Data
`
`US 2008/0262059 A1
`
`Oct. 23, 2008
`
`Related US, Application Data
`
`(62) Division of application No. 10/341,868, filed on Jan.
`14, 2003, now Pat. No. 7,468,390.
`.
`.
`.
`.
`Prov1s1onal applrcatron No. 60/386,792, filed on Jun.
`7, 2002, provisional application No. 60/349,660, filed
`on Jan. 175 2002.
`
`(60)
`
`(51)
`
`Int Cl
`(2006.01)
`A61K 31/235
`(2006.01)
`A61K 31/41
`(2006.01)
`A61K 31/195
`(52) us. Cl.
`......... 514/533, 514/381; 514/561; 514/563
`(58) Field of Classification Search .................. 514/533,
`514/381, 563
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`9/1986
`2/1988
`4/1988
`6/1988
`5/1990
`6/1993
`6/1993
`12/1993
`3/1994
`3/1995
`5/1996
`
`......... 549/452
`Berger .................
`260/402.5
`Delaney et al.
`
`..... 514/13
`Olins
`.. 514/19
`
`.
`Haslanger et
`..
`514/506
`Haslanger et a1.
`..
`
`Ksander ..........
`514/533
`.. 514/339
` De Lombaert .......
`Delaney et al.
`.
`514/381
`Erion et a1.
`......... 514/381
`514/381
`Buhlmayer et a1.
`Rathore et al.
`.......
`......... 417/322
`
`>>>>>>>>>>>
`
`4,610,816
`4,722,810
`4,740,499
`4,749,688
`4,929,641
`5,217,996
`5,223,516
`5,273,990
`5,294,632
`5,399,578
`5,520,522
`
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`GB
`W0
`W0
`W0
`W0
`
`FOREIGN PATENT DOCUMENTS
`0 342 850
`11/1989
`0 343 911
`11/1989
`0 361 365
`4/1990
`0 443 983
`8/1991
`0 498 361
`8/1992
`555175
`1/1993
`0 636 621
`2/1995
`0726072
`2/1996
`0 726 072
`8/1996
`0636621
`3/1997
`2 218 983
`11/1989
`90/09374
`8/1990
`92/14706
`9/1992
`93/09101
`5/1993
`93/10773
`6/1993
`
`B1
`
`$8
`WO
`WC
`WO
`
`3411/3223
`1(7);£33411
`02/06253
`“2002
`02/092622
`11/2002
`03/066606
`8/2003
`OTHER PUBLICATIONS
`
`Almeida et al., “Clearance Function of Type C Receptors of Atrial
`Natriuretic Factor in Rats”, Am J Physiol, vol. 256, pp. R469-R475
`(1989).
`EMU, Emlanfiméwebb, “Teéemettric M0?it0fiing if Cfldiofiicgla;
`arame eres 1n
`onscious
`pon aneous y
`ype ensrve
`s ,
`Cardiovasc Pharmacol, vol. 22, pp. 897-905 (1993).
`Consensus Trial Study Group, “Effects of Enalapril on Mortality in
`Severe Congestive Heart Failure”, NEng JMed, vol. 316, No. 23, pp.
`1429-1435(1987)
`Stephenson et al., The hydrolysis of a-human atrial natriuretic
`peptide by pig kindney microvillar membranes is initiated by
`endopeptidase-24.11 Biochem J, vol. 243, pp. 183-187(1987).
`Erdos, “Angiotensin 1 Converting Enzyme and the Changes in Our
`Concepts Through the Years”iLeWis K. Dahl Memorial Lecture,
`Hypertension, vol. 16, No. 4, pp. 363-370 (1990).
`Intengan, Park and Schiffrin, “Blood Pressure and Small Arteries in
`DOCA-Salt-Treated Genetically AVP-Deficient Rats”, Hyperten—
`sion, vol. 34, No. 4, Part 2, pp. 907-913 (1999).
`Intengan, Thibault, Li and Schiffrin, “Resistance Artery Mechanics,
`Structure,
`and Extracellular Components
`in Spontaneously
`Hypertensive Rats”, Circulation, vol, 100, No. 22, pp. 2267—2275
`(1999).
`Needleman et al., “The Biochemical Pharmacology of Atrial
`Peptides”, Annu Rev Pharm Tax, vol. 29, pp. 23-54 (1989).
`Stephenson and Kenny, “Metabolism ofNeuropeptides”, Biochem J,
`“’1' 241’ pp’ 237447 (1987)
`(Continued)
`.
`.
`Primary Exammer * Jenmfer M Km
`(74) Attorney, Agent. or Firm * Stephen E- Johnson
`
`(57)
`
`ABSTRACT
`
`The invention relates a pharmaceutical composition compris-
`ing a combination of:
`(i)
`the AT l-antagonist valsartan or a pharmaceutically
`acceptable salt thereof; and
`(ii) a NEP inhibitor or a pharmaceutically acceptable salt
`thereof and optionally a pharmaceutically acceptable car-
`rier and to a method for the treatment or prevention of a
`condition or disease
`selected from the group consisting of hypertension, heart
`failure, such as (acute and chronic) congestive heart failure,
`left ventricular dysfunction and hypertrophic cardiomyopa-
`thy, diabetic cardiac myopathy, supraventricular and ven-
`tricular arrhythmias, atrial fibrillation, atrial flutter, detrimen-
`tal vascular
`remodeling, myocardial
`infarction and its
`sequelae, atherosclerosis, angina (whether unstable or
`stable), renal insufficiency (diabetic and non-diabetic), heart
`failure, angina pectoris, diabetes, secondary aldosteronism,
`primary and secondary pulmonary hypertension, renal failure
`conditions, such as diabetic nephropathy, glomerulonephri-
`tis, scleroderma, glomerular sclerosis, proteinuria of primary
`renal disease, and also renal vascular hypertension, diabetic
`retinopathy, the management of other vascular disorders,
`such as migraine, peripheral vascular disease, Raynaud’ s dis-
`ease,
`luminal hyperplasia, cognitive dysfunction, such as
`Alzheimer’s, glaucoma and stroke, comprising administering
`a therapeutically effective amount of the pharmaceutical
`composition to a mammal in need thereof.
`
`4 Claims, N0 Drawings
`
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`

`

`US 8,101,659 B2
`Page 2
`
`OTHER PUBLICATIONS
`
`Sybertz et a1., “SCH 39370, a Neutral Metalloendopeptidase Inhibi-
`tor, Potentiates Biological Responses to Atrial Natriuretic Factor and
`Lowers Blood Pressure in Desoxycorticosterone Acetate-Sodium
`Hypertensive Rats”, JPharmacol Exp Ther, vol. 250, No. 2, pp.
`624-631 (1989).
`and
`“Atrial Natriuretic Factor-Potentiating
`Sybertz
`et
`a1.,
`Antihypertensive Activity of SCH 34826”, Hypertension, vol. 15,
`No. 2, pp. 152-161 (1990).
`Williford, Sharma, Korth and Sheu, “Spatial Heterogeneity of
`Intracellular Ca2+ Concentration in Nonbeating Guinea Pig
`Ventricular Myocytes”, Circ Res, vol. 66, No. 1, pp. 241-249 (1990).
`Zannad, “The Emerging Role of ACE Inhibitors in the Treatment of
`Cardiovascular Disease”, J Cardiovasc Pharmacol, vol. 15, Suppl. 2,
`pp. Sl, S5 (1990).
`CAPLUS Abstract AN 1986:5730427Taub et a1., f ZA8400670,
`Sep. 25, 1985.
`CAPLUS Abstract AN 1995: 9312307Sugano et a1., JP 07157459,
`Jun. 20, 1995.
`CAPLUS Abstract AN 1995:4126607Yamada et a1., JP 06234754,
`Aug. 23, 1994.
`Matsumoto et al. “Blockade of renin-angiotensin system and
`enhancement of atrial natriuretic peptide with neutral endopedtidase
`inhibition cause natriuresis in congestive heart failure and renal dys-
`function in conscious dogs”. JASN, Hemodynamics and Vascular
`Regulation, pp. 517, (1993).
`Lajemi et a1., “Genetics of the renin-angiotensin-aldosterone system
`and risk of arterial disease”; ACE Inhibitors, Milestones in Drug
`Therapy, pp. 11-27; P. D’Orleans-Juste, G.E. Plante (2001).
`
`Matsumoto et a1., “Blockade of renin-angiotensin system and
`enhancement of atrial natriuretic peptide with neutral endopeptidase
`inhibition cause natriuresis in congestive heart failure and renal dys-
`function in conscious dogs”, ASN Program and Abstracts, vol. 4, No.
`3, pp. 517( Sep. 1993).
`Robi et a1., “Neutral endopeptidase inhibitors and combined inhibi-
`tors of neutral endopeptidase and angiotensin converting enzyme”,
`Antihypersensitive Drugs,pp. 113-212 , (1997).
`Wohlfart, et a1., “Crosstalk between ACE Inhibitors, B2 kinin recep-
`tor and nitric oxide in endothelial cells”, ACE Inhibitors, Milestones
`in Drug Therapy, P. D’Orleans-Juste, G.E. Plante (2001).
`Trippodo et a1., Repression of Angiotensin II and Potentiation of
`Bradykinin Contribute to the Synergistic Effects of Dual Metal-
`loprotease Inhibition in Heart Failure:, J. Pharmacol. Exp. Ther., vol.
`272, pp. 619-627, (1995).
`Howes et a1., “Angiotensin receptor antagonists and ACE inhibitors”,
`Australian Family Physician, vol. 27, pp. 914-921, (1998).
`Criscione et a1., “Pharmacological profile of valsartan: a potent,
`orally active, nonpeptide antagonists of the angiotensin II AT-recep-
`tor sybtype”, Br. J. Pharmacol., vol. 110, pp. 761-771, (1993).
`Abstract of J. Pharmacol. Exp., Ther., vol. 265, pp. 1339-1347,
`(1993).
`Search dated May 29, 2008 (26 pages) cited by opponent
`Mundipharma GMBH on Jun. 26, 2008, submitted in corresponding
`EP application 03704413.8.
`Search dated May 29, 2008 (3 pages) cited by opponent
`Mundipharma GMBH on Jun. 26, 2008, submitted in corresponding
`EP application 03704413.8.
`
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`

`

`US 8,101,659 B2
`
`1
`METHODS OF TREATMENT AND
`PHARMACEUTICAL COMPOSITION
`
`This application is a continuation application of US.
`patent application Ser. No. 10/341,868 filed on Jan. 14, 2003
`and claims benefit of US. Provisional Pat. Appl. No. 60/386,
`792, filed Jun. 7, 2002 and US. Provisional Pat. Appl. No.
`60/349,660, filed Jan. 17, 2002, the entire disclosures of
`which are hereby incorporated by reference.
`
`BACKGROUND OF THE INVENTION
`
`The renin angiotensin system is a complex hormonal sys-
`tem comprised of a large molecular weight precursor, angio-
`tensinogen, two processing enzymes, renin and angiotensin
`converting enzyme (ACE), and the vasoactive mediator
`angiotensin II (Ang 11). See J Cardiovasc. Pharmacol., Vol.
`15, Suppl. B, pp. 81-85 (1990). The enzyme renin catalyzes
`the cleavage of angiotensinogen into the decapeptide angio-
`tensin l, which has minimal biological activity on its own and
`is converted into the active octapeptide Ang II by ACE. Ang II
`has multiple biological actions on the cardiovascular system,
`including vasoconstriction, activation ofthe sympathetic ner-
`vous system, stimulation of aldosterone production, anti-
`natriuresis, stimulation of vascular growth and stimulation of
`cardiac growth. Ang 11 functions as a pressor hormone and is
`involved the pathophysiology of several forms of hyperten-
`s10n.
`
`The vasoconstrictive effects of angiotensin II are produced
`by its action on the non-striated smooth muscle cells, the
`stimulation of the formation of the adrenergenic hormones
`epinephrine and norepinephrine, as well as the increase ofthe
`activity of the sympathetic nervous system as a result of the
`formation of norepinephrine. Ang II also has an influence on
`electrolyte balance, produces, e.g., anti-natriuretic and anti-
`diuretic effects in the kidney and thereby promotes the release
`of, on the one hand, the vasopres sin peptide from the pituitary
`gland and, on the other hand, of aldosterone from the adrenal
`glomerulo sa. All these influences play an important part in the
`regulation of blood pressure, in increasing both circulating
`volume and peripheral resistance. Ang II is also involved in
`cell growth and migration and in extracellular matrix forma-
`tion.
`
`Ang II interacts with specific receptors on the surface ofthe
`target cell. It has been possible to identify receptor subtypes
`that are termed, e.g., AT 1- andAT 2-receptors. In recent times
`great efforts have been made to identify substances that bind
`to the AT 1-receptor. Such active ingredients are often termed
`Ang II antagonists. Because of the inhibition of the AT 1-re-
`ceptor such antagonists can be used, e.g., as anti-hyperten-
`sive’s or for the treatment of congestive heart failure, among
`other indications.Ang H antagonists are therefore understood
`to be those active ingredients which bind to the AT 1-receptor
`subtype.
`Inhibitors of the renin angiotensin system are well-known
`drugs that lowerblood pressure and exert beneficial actions in
`hypertension and in congestive heart failure as described.
`See, e.g, N. Eng. J. Med., Vol. 316, No. 23, pp. 1429-1435
`(1987). A large number of peptide and non-peptide inhibitors
`of the renin angiotensin system are known, the most widely
`studied being the ACE inhibitors, which includes the drugs
`captopril, enalapril,
`lisinopril, benazepril and spirapril.
`Although a major mode of action ofACE inhibitors involves
`prevention offormation ofthe vasoconstrictor peptide Ang II,
`it has been reported in Hypertension, Vol. 16, No. 4, pp.
`363-370 (1990), that ACE cleaves a variety of peptide sub-
`strates, including the vasoactive peptides bradykinin and sub-
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`2
`
`stance P. Prevention ofthe degradation ofbradykinin by ACE
`inhibitors has been demonstrated, and the activity of the ACE
`inhibitors in some conditions has been reported in Circ. Res.,
`Vol. 66, No. 1, pp. 242-248 (1990), to be mediated by eleva-
`tion of bradykinin levels rather than inhibition of Ang II
`formation. Consequently,
`it cannot be presumed that the
`effect of an ACE inhibitor is due solely to prevention of
`angiotensin formation and subsequent inhibition of the renin
`angiotensin system.
`Neutral endopeptidase (EC 3.4.24.11; enkephalinase;
`atriopeptidase; NEP) is a zinc-containing metalloprotease
`that cleaves a variety of peptide substrates on the amino
`terminal side of aromatic amino acids. See Biochem. J., Vol.
`241, pp. 237-247 (1987). Substrates for this enzyme include,
`but are not limited to, atrial natriuretic factors (ANFs), also
`known as ANPs, brain natriuretic peptide (BNP), met and leu
`enkephalin, bradykinin, neurokinin A and substance P.
`ANPs are a family of vasodilator, diuretic and anti-hyper-
`tensive peptides which have been the subject of many recent
`reports inthe literature. See, e.g.,Annu. Rev. Pharm. Tox.,Vol.
`29, pp. 23-54 (1989). One form, ANF 99-126, is a circulating
`peptide hormone which is released from the heart during
`conditions of cardiac distension. The function of ANF is to
`
`maintain salt and water homeostasis, as well as to regulate
`blood pressure. ANF is rapidly inactivated in the circulation
`by at least two processes: a receptor-mediated clearance
`reported in Am. J Physiol., Vol. 256, pp. R469-R475 (1989),
`and an enzymatic inactivation via NEP reported in Biochem.
`J., Vol. 243, pp. 183-187 (1987). It has been previously dem-
`onstrated that inhibitors of NEP potentiate the hypotensive,
`diuretic, natriuretic and plasma ANF responses to pharmaco-
`logical injection ofANF in experimental animals. The poten-
`tiation of ANF by two specific NEP inhibitors is reported by
`Sybertz et al., J. Pharmacol. Exp. Ther., Vol. 250, No. 2, pp.
`624-631 (1989), and in Hypertension, Vol. 15, No. 2, pp.
`152-161 (1990), while the potentiation of ANF by NEP in
`general was disclosed in US. Pat. No. 4,749,688. In US. Pat.
`No. 4,740,499, Olins disclosed the use of thiorphan and kela-
`torphan to potentiate atrial peptides. Moreover, NEP inhibi-
`tors lower blood pressure and exert ANF-like effects, such as
`diuresis and increased cyclic guanosine 3',5'-monophosphate
`(cGMP) excretion in some forms of experimental hyperten-
`sion. The anti-hypertensive action of NEP inhibitors is medi-
`ated through ANF because antibodies to ANF will neutralize
`the reduction in blood pressure.
`Darrow et al. in European Patent Application No. 498361
`disclose treating hypertension or congestive heart failure with
`a combination of certain Ang II antagonists or certain renin
`inhibitors with certain NEP inhibitors.
`
`Powell et al. in European Patent Application No. 726072
`disclose treating hypertension or congestive heart failure with
`a combination of the Ang II antagonist 2-butyl-6,7,8,9-tet-
`rahydro-3-[[2'-(1H-tetrazol-5 -yl) [1 , 1 '-biphenyl] -4 -yl]me-
`thyl]-1,3-diazaspiro[4.4]nonan-4-one with a NEP inhibitor
`or a dual acting vasopeptidase inhibitor (single molecular
`entity with both ACE and NEP inhibitory activities). Pro-
`longed and uncontrolled hypertensive vascular disease ulti-
`mately leads to a variety of pathological changes in target
`organs, such as the heart and kidney. Sustained hypertension
`can lead as well to an increased occurrence of stroke. There-
`
`fore, there is a strong need to evaluate the efficacy of anti-
`hypertensive therapy, an examination ofadditional cardiovas-
`cular endpoints, beyond those of blood pressure lowering, to
`get further insight into the benefits of combined treatment.
`The nature of hypertensive vascular diseases is multifac-
`torial. Under certain circumstances, drugs with different
`mechanisms of action have been combined. However, just
`
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`

`US 8,101,659 B2
`
`3
`considering any combination of drugs having different mode
`of action does not necessarily lead to combinations with
`advantageous effects. Accordingly, there is a need for more
`efficacious combination therapy which has less deleterious
`side effects.
`Other objects, features, advantages and aspects of the
`present invention will become apparent to those of skill from
`the following description. It should be understood, however,
`that the following description and the specific examples,
`while indicating preferred embodiments of the invention, are
`given by way of illustration only. Various changes and modi-
`fications within the spirit and scope ofthe disclosed invention
`will become readily apparent to those skilled in the art from
`reading the following description and from reading the other
`parts of the present disclosure.
`
`DETAILED DESCRIPTION OF THE PREFERRED
`EMBODIMENTS
`
`In one aspect, the present invention relates to pharmaceu-
`tical combinations comprising valsartan or pharmaceutically
`acceptable salts thereof and a NEP inhibitor or a pharmaceu-
`tically effective salts thereof, optionally in the presence of a
`pharmaceutically acceptable carrier and pharmaceutical
`compositions comprising them.
`In another embodiment, the present invention relates to
`methods of treating cardiac and renal related conditions by
`administration of the pharmaceutical composition compris-
`ing valsartan plus a NEP inhibitor.
`Valsartan is the AT 1-receptor antagonist (S)iN-(1-car-
`boxy-2 -methyl-prop- 1 -yl) -N-pentanoyl-N— [2;
`(1H-tetrazol-
`5-yl)biphenyl-4-yl-methyl] amine of formula (I)
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`4
`
`R3 is hydrogen, alkyl of 1 to 7 carbons, phenyl, substituted
`phenyl, 7(CH2)1 to 4-phenyl, or 7(CH2)1 to 4-substi-
`tuted phenyl;
`R1 is hydroxy, alkoxy of 1 to 7 carbons, or NH2;
`11 is an integer from 1 to 15; and
`the term substituted phenyl refers to a substituent selected
`from lower alkyl of 1 to 4 carbons, lower alkoxy of 1 to 4
`carbons, lower alkylthio of 1 to 4 carbons, hydroxy, Cl, Br or
`F.
`
`Preferred selective NEP inhibitors of formula (II) include
`compounds, wherein
`R2 is benzyl;
`R3 is hydrogen;
`n is an integer from 1 to 9; and
`R1 is hydroxy.
`Even more preferred selective NEP inhibitors of formula
`(II) are reported in the literature as SQ 28,603 which is the
`compound of formula (II),
`wherein
`
`R2 is benzyl;
`R3 is hydrogen;
`n is one; and
`R1 is hydroxy.
`The preparation of the selective NEP inhibitors of formula
`(II), wherein R2 is other than trifluoromethyl are disclosed by
`Delaney et al. in US. Pat. No. 4,722,810. The preparation of
`the selective NEP inhibitors of formula (II), wherein R2 is
`trifluoromethyl are disclosed by Delaney et al. in US. Pat.
`No. 5,223,516.
`NEP inhibitors within the scope of the present invention
`include compounds disclosed in US. Pat. No. 4,610,816,
`herein incorporated by reference,
`including in particular
`Ni[N-[1(S)-carboxyl-3 -phenylproplyl] -(S)-phenylalanyl] -
`(S)-isoserine and Ni[N-[((1S)-carboxy-2-phenyl)ethyl]-
`(S)-phenylalanyl]-[3-alanine; compounds disclosed in US.
`Pat. No. 4,929,641, in particular, N—[2(S)-mercaptomethyl-3-
`(2-methylphenyl)-propionyl]methionine; SQ 28603 (N-[2-
`(mercaptomethyl)-1-oxo-3-phenylpropyl]-[3-alanine),
`dis-
`closed in South African Patent Application No. 84/0670; UK
`69578 (cis-4-[[[1-[2-carboxy-3-(2-methoxyethoxy)propyl]-
`cyclopentyl]carbonyl]amino]-cyclohexanecarboxylic
`acid)
`and its active enantiomer(s); thiorphan and its enantiomers;
`retro-thiorphan; phosphoramidon; and SQ 29072 (7-[[2-
`(mercaptomethyl) -1 -oxo -3 -phenylpropyl] amino] -heptanoic
`acid). Also suitable for use are any pro-drug forms of the
`above-listed NEP inhibitors, e.g., compounds in which one or
`more carboxylic acid groups are esterified.
`NEP inhibitors within the scope of the present invention
`also include the compounds disclosed in US. Pat. No. 5,217,
`996, particularly, N—(3-carboxy-1-oxopropyl)-(4S)-p-phe-
`nylphenylmethyl)-4-amino-2R-methylbutanoic acid ethyl
`ester; the compounds disclosed in EP 00342850, particularly,
`(S)-cis-4-[1-[2-(5-indanyloxycarbonyl)-3-(2-methoxy-
`ethoxy)propyl]—1-cyclopentanecarboxamido]-1 -cyclohexan-
`ecarboxylic acid; the compounds disclosed in GB 02218983,
`particularly, 3-(1-[6-endo-hydroxymethylbicyclo[2,2,1]hep-
`tane-2-exo-carbamoyl]cyclopentyl)-2-(2-methoxyethyl)pro-
`panoic acid; the compounds disclosed in W0 92/ 14706, par-
`ticularly, N—(1-(3-(N—t-butoxycarbonyl-(S)-prolylamino)-2
`(S)-t-butoxy-carbonylpropyl)cyclopentanecarbonyl)-O-
`benzyl-(S)-serine methyl ester; the compounds disclosed in
`EP 00343911; the compounds disclosed in JP 06234754; the
`compounds disclosed in EP 00361365, particularly, 4-[[2-
`(mercaptomethyl) -1 -oxo -3 -phenylpropyl] amino]benzoic
`acid; the compounds disclosed in WO 90/09374, particularly,
`3 -[(1-(cis-4-carboxycarbonyl-cis-3 -butylcyclohexyl-r-1-car-
`boamoyl)cyclopentyl]-2S-(2-methoxyethoxymethyl)pro-
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`CH
`CH3\ / 2\
`CH2
`
`CH3
`
`CH3\
`(1 Hi/
`C
`C
`CH2/ \N/HYOH
`
`(1)
`
`35
`
`0
`
`\N
`HN
`
`\ /N=N
`
`and is disclosed in EP 0443983 A and US. Pat. No. 5,399,
`578, the disclosures of which are incorporated herein in their
`entirety as if set forth herein.
`A NEP inhibitor useful in said combination is a compound
`of the formula (II)
`
`R3
`0
`R2
`II
`I
`II
`I
`Hs—CHz—CH—C—NH—CH—(CH2),.—C—Rl
`
`0
`
`(H)
`
`and pharmaceutically acceptable salts thereof,
`wherein
`
`R2 is alkyl of 1 to 7 carbons, trifluoromethyl, phenyl, sub-
`stituted phenyl, 7(CH2)1 to 4-phenyl, or 7(CH2)1 to 4-
`substituted phenyl;
`
`BIOCON PHARMA LTD (IPR2020-01263) EX. 1001, p. 004
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`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1001, p. 004
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`

`US 8,101,659 B2
`
`5
`panoic acid; the compounds disclosed in JP 07157459, par-
`ticularly,
`N—((2S)-2-(4-biphenylmethyl)-4-carboxy-5-
`phenoxyvaleryl)glycine; the compounds disclosed in WO
`94/15908, particularly, N—(1-(N—hydroxycarbamoylmethyl)-
`1-cyclopentanecarbonyl)-L-phenylalanine;
`the compounds
`disclosed in U.S. Pat. No. 5,273,990, particularly, (S)-(2-
`biphenyl-4-yl)-1 -(1H-tetrazol-5-yl)ethylamino)
`meth-
`ylphosphonic acid; the compounds disclosed in U.S. Pat. No.
`5,294,632,
`particularly,
`(S)-5-(N—(2-(phosphonomethy-
`lamino)-3-(4-biphenyl)propionyl)-2-aminoethyl)tetrazole;
`the compounds disclosed in US. Pat. No. 5,250,522, particu-
`larly, B-Alanine, 3-[1,1'-biphenyl]-4-yl-N-[diphenoxyphos-
`phinyl)methyl]—L-alanyl;
`the compounds disclosed in EP
`00636621,
`particularly, N—(2-carboxy-4-thienyl)-3-mer-
`capto-2-benzylpropanamide;
`the compounds disclosed in
`WO 93/09101, particularly, 2-(2-mercaptomethyl-3-phenyl-
`propionamido)thiazol-4-ylcarboxylic acid; the compounds
`disclosed in EP 00590442, particularly, ((L)-(1-((2,2-dim-
`ethyl-1 ,3 -dioxolan—4-yl) -methoxy)carbonyl)-2-phenyl-
`ethyl)-L-phenylalanyl)-[3-alanine, Ni[N-[(L)-[1-[(2,2-dim-
`ethyl-1,3 -dioxolan—4-yl) -methoxy]carbonyl] -2-
`phenylethyl] -L-phenylalanyl] -(R)-alanine, Ni[N-[(L)-1-
`carboxy-2-phenylethyl]-L-phenylalanyl]-(R)-alanine, N—[2-
`acetylthiomethyl-3-(2-methyl-phenyl)propionyl]-
`methionine
`ethyl
`ester,
`N—[2-mercaptomethyl-3-(2-
`methylphenyl)propioyl] -methionine,
`N—[2(S)-
`mercaptomethyl-3 - (2 -methylphenyl)propanoyl] -(S) -
`isoserine,
`N7(S)-[3-mercapto-2-(2-methylphenyl)
`propionyl] -(S)-2 -methoxy- (R) -alanine,
`N-[1-[[1(S)-
`benzyloxycarbonyl-3-phenylpropyl]amino]
`cyclopentylcarbonyl](S)-isoserine, N—[1-[[1(S)-carbonyl-3-
`phenylpropy] amino] -cyclopentylcarbonyl] -(S)-isoserine,
`1,1'-[dithiobis-[2(S)-(2-methylbenzyl)-1-oxo-3,1-pro-
`panediyl]]-bis-(S)-isoserine,
`1,1'-[dithiobis-[2(S)-(2-meth-
`ylbenzyl)-1-oxo-3,1-propanediyl]]-bis-(S)-methionine,
`N—(3-phenyl-2-(mercaptomethyl)-propionyl)-(S)-4-(meth-
`ylmercapto)methionine, N—[2-acetylthiomethyl-3-phenyl-
`propionyl]-3-aminobenzoic acid, N—[2-mercaptomethyl-3-
`phenyl-propionyl]-3-aminobenzoic acid, N—[1-(2-carboxy-
`4-phenylbutyl)-cyclopentanecarbonyl]-(S)-isoserine, N—[1-
`(acetylthiomethyl)cyclopentane-carbonyl] -(S) -methionine
`ethyl ester, 3(S)-[2-(acetylthiomethyl)-3-phenyl-propionyl]
`amino-e-caprolactam; and the compounds disclosed in WO
`93/10773, particularly, N—(2-acetylthiomethyl-3-(2-meth-
`ylphenyl)propionyl)-methionine ethyl ester.
`The compounds to be combined can be present as pharma-
`ceutically acceptable salts. If these compounds have, for
`example, at least one basic center, they can form acid addition
`salts. Corresponding acid addition salts can also be formed
`having, if desired, an additionally present basic center. The
`compounds having at least one acid group, for example,
`COOH, can also form salts with bases. Corresponding inter-
`nal salts may furthermore be formed, if a compound com-
`prises, e.g., both a carboxy and an amino group.
`With respect to N-(3-carboxy-1-oxopropyl)-(4S)-p-phe-
`nylphenylmethyl)-4-amino-2R-methylbutanoic acid ethyl
`ester, preferred salts include the sodium salt disclosed in US.
`Pat. No. 5,217,996, the triethanolamine salt and the tris(hy-
`droxymethyl)aminomethane salt. Preparation of the trietha-
`nolamine salt and the tris(hydroxymethyl)aminomethane salt
`may be carried out as follows:
`Triethanolamine
`
`To N-(3 -carboxy-1-oxopropyl)-(4S)-p-phenylphenylm-
`ethyl)-4-amino-2R-methylbutanoic acid ethyl ester (349 mg,
`0.848 mmol) is added 5 mL of ethyl ether and 0.113 mL
`(0.848 mmol) oftriethanolamine in 1 mL ofethyl acetate. The
`solid was collected and dried melting at 69-710 C.
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`HO
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`Tris(hydroxymethyl)aminomethane
`To N-(3-carboxy-1-oxopropyl)-(4S)-p-phenylphenylm-
`ethyl)-4-amino-2R-methylbutanoic acid ethyl ester (3.2 g,
`7.78 mmol) is added 32 mL ofethyl acetate and 940 mg (7.78
`mmol) tris(hydroxymethyl)aminomethane. The suspension
`is diluted with 45 mL of ethyl acetate and refluxed overnight
`(~20 hours). The reaction is cooled to 0° C., filtered, solid
`washed with ethyl acetate and dried melting at 114-1150 C.
`It has surprisingly been found that, a combination of val-
`sartan and a NEP inhibitor achieves greater therapeutic effect
`than the administration of valsartan, ACE inhibitors or NEP
`inhibitors alone and promotes less angioedema than is seen
`with the administration of a vasopeptidase inhibitor alone.
`Greater efficacy can also be documented as a prolonged dura-
`tion of action. The duration of action can be monitored as
`
`either the time to return to baseline prior to the next dose or as
`the area under the curve (AUC) and is expressed as the prod-
`uct of the change in blood pressure in millimeters of mercury
`(change in mmHg) and the duration of the effect (minutes,
`hours or days).
`Further benefits are that lower doses ofthe individual drugs
`to be combined according to the present invention can be used
`to reduce the dosage, for example, that the dosages need not
`only often be smaller but are also applied less frequently, or
`can be used to diminish the incidence of side effects. The
`
`combined administration of valsartan or a pharrnaceutically
`acceptable salt thereof and a NEP inhibitor or a pharmaceu-
`tically acceptable salt thereof results in a significant response
`in a greater percentage of treated patients, that is, a greater
`responder rate results, regardless of the underlying etiology
`of the condition. This is in accordance with the desires and
`
`requirements of the patients to be treated.
`It can be shown that combination therapy with valsartan
`and a NEP inhibitor results in a more effective anti -hyperten-
`sive therapy (whether for malignant, essential, reno -vascular,
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1001, p. 005
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`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1001, p. 005
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`US 8,101,659 B2
`
`7
`diabetic, isolated systolic or other secondary type of hyper-
`tension) through improved efiicacy, as well as a greater
`responder rate. The combination is also useful in the treat-
`ment or prevention of heart failure, such as (acute and
`chronic) congestive heart failure, left ventricular dysfunction
`and hypertrophic cardiomyopathy, diabetic cardiac myopa-
`thy, supraventricular and ventricular arrhythmias, atrial fibril-
`lation, atrial flutter or detrimental vascular remodeling. It can
`further be shown that a valsartan and NEP inhibitor therapy
`proves to be beneficial in the treatment and prevention of
`myocardial infarction and its sequelae. A valsartan plus NEP
`inhibitor combination is also useful in treating atherosclero-
`sis, angina (whether stable or unstable), and renal insufii-
`ciency (diabetic and non-diabetic). Furthermore, combina-
`tion therapy using valsartan and a NEP inhibitor can improve
`endothelial dysfunction, thereby providing benefit in diseases
`in which normal endothelial function is disrupted, such as
`heart failure, angina pectoris and diabetes. Furthermore, the
`combination of the present invention may be used for the
`treatment or prevention of secondary aldosteronism, primary
`and secondary pulmonary hypertension, renal failure condi-
`tions, such as diabetic nephropathy, glomerulonephritis, scle-
`roderrna, glomerular sclerosis, proteinuria of primary renal
`disease and also renal vascular hypertension, diabetic retin-
`opathy, the management of other vascular disorders, such as
`migraine, peripheral vascular disease, Raynaud’s disease,
`luminal hyperplasia, cognitive dysfunction, such as Alzhe-
`imer’s; glaucoma and stroke.
`The person skilled in the pertinent art is fully enabled to
`select a relevant test model to prove the efficacy of a combi-
`nation of the present invention in the hereinbefore and here-
`inafter indicated therapeutic indications.
`Representative studies are carried out with a combination
`of valsartan and N—(3-carboxy-1-oxopropyl)-(4S)-p-phe-
`nylphenylmethyl)-4-amino-2R-methylbutanoic acid ethyl
`ester, e.g., applying the following methodology:
`Drug efficacy is assessed in various animal models includ-
`ing the deoxycorticosterone acetate-salt (DOCA-salt) rat and
`the spontaneously hypertensive rat (SHR), either maintained
`on a normal salt diet or with salt loading (4-8% salt in rat chow
`or 1% NaCl as drinking water).
`The DOCA-salt test model utilizes either an acute or
`
`chronic study protocol. An acute study procedure involves
`assessment of the effects of various test substances over a
`
`six-hour experimental period using rats with indwelling
`femoral arterial and venous catheters. The acute study proce-
`dure evaluates test substances for their ability to reduce blood
`pressure during the established phase of DOCA-salt hyper-
`tension. In contrast, the chronic study procedure assesses the
`ability of test substances to prevent or delay the rise in blood
`pressure during the development phase of DOCA-salt hyper-
`tension. Therefore, blood pressure will be monitored in the
`chronic study procedure by means of a radiotransmitter. The
`radiotransmitter is surgically implanted into the abdominal
`aorta of rats, prior to the initiation of DOCA-salt treatment
`and thus, prior to the induction of hypertension. Blood pres-
`sure is chronically monitored for periods of up to six weeks
`(approximately one week prior to DOCA-salt administration
`and for five weeks thereafter).
`Rats are anesthetized with 2-3% isoflurane in oxygen
`inhalant followed by Amytal sodium (amobarbital) 100
`mg/kg, i.p. The level of anesthesia is assessed by a steady
`rhythmic breathing pattern.
`Acute Study Procedure:
`Rats undergo a unilateral nephrectomy at the time of
`DOCA implantation. Hair is clipped on the left flank and the
`back of the neck and scrubbed with sterile alcohol swabs and
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`8
`povidone/iodine. During surgery rats are placed on a heating
`pad to maintain body temperature at 37° C.
`A 20 mm incision is made through the skin and underlying
`muscle to expose the left kidney. The kidney is freed of
`surrounding tissue, exteriorized and two ligatures (3-0 silk)
`are tied securely around the renal artery and vein proximal to
`theirj uncture with the aorta. The renal artery and vein are then
`severed and the kidney removed. The muscle and skin
`wounds are closed with 4-0 silk suture and stainless steel
`
`wound clips, respectively. At the same time, a 15 mm incision
`is made on the back of the neck and a three-week-release
`
`pellet (Innovative Research of America, Sarasota, Fla.) con-
`taining DOCA (100 mg/kg) is implanted subcutaneously
`(s.c.). The wound is then closed with stainless-steel clips and
`both wounds are treated with povidone/iodine; the rats are
`given a post-surgical
`intramuscular
`(i .m.)
`injection of
`procaine penicillin G (100,000 U) and buprenorphine (0.05-
`0.1 mg/kg) s.c. The rats are immediately placed on 1% NaCl+
`0.2% KCl drinking water; this treatment continues for at least
`3 weeks at which time the animals have become hypertensive
`and available for experimentation.
`Forty-eight hours prio