USOO7468390B2
`
`(12) United States Patent
`KSander et al.
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 7.468,390 B2
`Dec. 23, 2008
`
`(54) METHODS OF TREATMENT AND
`PHARMACEUTICAL COMPOSITION
`
`G Michael Ksander. Amherst, NH
`t
`(75) I
`nventors: Urary VIIcnaei Ksander,
`erSl,
`(US); Randy Lee Webb, Flemington, NJ
`(US)
`
`WO
`WO
`
`WO
`WO
`
`WOO1,74348 A2 10, 2001
`WOO2.06253
`1, 2002
`
`WO O2/O92622 A2 11/2002
`WO 03/066606
`8, 2003
`
`(73) Assignee: Novartis AG, Basel (CH)
`
`OTHER PUBLICATIONS
`
`(*) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 317 days.
`(21) Appl. No.: 10/341,868
`
`(22) Filed:
`
`Jan. 14, 2003
`
`(65)
`
`Prior Publication Data
`
`US 2003/O144215A1
`Jul. 31, 2003
`Related U.S. Application Data
`(60) Provisional application No. 60/386,792, filed on Jun.
`7, 2002, provisional application No. 60/349,660, filed
`on Jan. 17, 2002.
`
`(51) Int. Cl.
`(2006.01)
`A6 IK 3L/235
`(2006.01)
`A6 IK3I/41
`(2006.01)
`A 6LX 3L/95
`(52) U.S. Cl. ....................... 514/533; 514/381: 514/561;
`514/563
`(58) Field of Classification Search ................. 514/533,
`514/381,561,563
`See application file for complete search history.
`References Cited
`
`(56)
`
`Almeida et al., "Clearance Function of Type C Receptors of Atrial
`Natriuretic Factor in Rats'. Am J Physiol, vol. 256, pp. R469-R475
`(1989).
`Bazil, Krulan and Webb, "Telemetric Monitoring of Cardiovascular
`Parameteres in Conscious Spontaneously Hypertensive Rats', J
`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
`eptide by pig kindnev microvillar membranes is initiated b
`Sail" E. J. vol. 243, pp. 183-187(1987).
`y
`Erdös, "Angiotensin I Converting Enzyme and the Changes in Our
`Concepts Through the Years' Lewis K. Dahl Memorial Lecture,
`Hypertension, vol. 16, No. 4, pp. 363-370 (1990).
`
`(Continued)
`Primary Examiner Jennifer Kim
`(74) Attorney, Agent, or Firm Gregory D. Ferraro
`
`(57)
`
`ABSTRACT
`
`The invention relates a pharmaceutical composition compris
`ing a combination of
`(i) the AT 1-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 fail
`
`omyopathy, diabetic cardiac myopathy, Supraventricular
`and ventricular arrhythmias, atrial fibrillation, atrial flutter,
`detrimental 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 aldoster
`onism, primary and secondary pulmonary hypertension,
`renal failure conditions, such as diabetic nephropathy,
`glomerulonephritis, Scleroderma, glomerular Sclerosis,
`proteinuria of primary renal disease, and also renal vascu
`lar hypertension, diabetic retinopathy, the management of
`other vascular disorders, such as migraine, peripheral vas
`cular disease, Raynaud's disease, luminal hyperplasia,
`cognitive dysfunction, such as Alzheimers, glaucoma and
`stroke, comprising administering a therapeutically effec
`tive amount of the pharmaceutical composition to a mam
`mal in need thereof.
`
`3 Claims, No Drawings
`
`E. E. O.E. ure, left venticular dysfunction and hypertrophic cardi
`
`U.S. PATENT DOCUMENTS
`4,610,816 A
`9/1986 Berger ........................ 549,452
`4,722,810 A
`2/1988 Delaney et al. .
`... 260/402.5
`4,740,499 A
`4, 1988 Olins .............
`... 514f13
`4,749,688 A
`6/1988 Haslanger et al. ............. 514/19
`4,929,641 A
`5/1990 Haslanger et al. ........... 514,506
`
`E. R.
`
`5,273.990 A 12/1993 De Lombaert .............. 514,381
`5,294.633 A
`3/1994 Erion et al. ................. 514,381
`5,399.578 A * 3/1995 Buhlmayer et al. ......... 514,381
`5,520,522 A
`5/1996 Rathore et al. .............. 417.322
`
`
`
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`GB
`W
`WO
`WO
`WO
`
`FOREIGN PATENT DOCUMENTS
`O 342 850
`11, 1989
`O 343 911
`11, 1989
`O 361 365
`4f1990
`O 443 983
`8, 1991
`O 498 361 A2
`8, 1992
`O 636 621
`2, 1995
`O 726 OT2 A2
`8, 1996
`O636621 B1
`3/1997
`2218983
`11, 1989
`W E.
`1.
`WO93,09101
`5, 1993
`WO 93/10773
`6, 1993
`WO94, 15908
`T 1994
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1014, p. 001
`
`

`

`US 7,468.390 B2
`Page 2
`
`OTHER PUBLICATIONS
`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., “Biochemical Pharmacology of Atrial Peptides'.
`Annu Rev Pharm Tox, vol. 29, pp. 23-54 (1989).
`Stephenson and Kenny, "Metabolism of Neuropeptides'. Biochem J.
`vol. 241, pp. 237-247 (1987).
`Sybertz et al., "SCH 39370, a Neutral Metalloendopeptidase Inhibi
`tor, Potentiates Biological Responses to Atrial Natriuretic Factor and
`Lowers Blood Pressure in Desoxycorticosterone Acetate-Sodium
`Hypertensive Rats', J Pharmacol Exp. Ther, vol. 250, No. 2, pp.
`624-631 (1989).
`Sybertz et al., “Atrial Natriuretic Factor-Potentiating and
`Antihypertensive Activity of SCH 34826'. Hypertension, vol. 15.
`No. 2, pp. 152-161 (1990).
`
`Williford, Sharma, Korth and Sheu, "Spatial Heterogeneity of
`Intracellular Ca' 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. S1, S5 (1990).
`Caplus Abstract AN 1986:573042 Taub et al., f ZA8400670, Sep.
`25, 1985.
`Caplus Abstract AN 1995:931230- Sugano et al., JP 07157459, Jun.
`20, 1995.
`Caplus Abstract AN 1995:412660 Yamada et al., JP 06234754,
`Aug. 23, 1994.
`Matsumoto et al., “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'. JASN, Hemodynamics and Vascular
`Regulation, p. 517. Sep.1993).
`* cited by examiner
`
`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1014, p. 002
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`

`

`US 7,468.390 B2
`
`1.
`METHODS OF TREATMENT AND
`PHARMACEUTICAL COMPOSITION
`
`BACKGROUND OF THE INVENTION
`
`2
`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 in the 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 of ANF 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 U.S. Pat. No. 4,749,688. In U.S. 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-diazaspiro4.4 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 of additional 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
`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.
`
`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 II). See J. Cardiovasc. Pharmacol., Vol.
`15, Suppl. B. pp. S1-S5 (1990). The enzyme renin catalyzes
`the cleavage of angiotensinogen into the decapeptide angio
`tensin I, 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 of the sympathetic ner
`Vous system, stimulation of aldosterone production, anti
`natriuresis, stimulation of vascular growth and stimulation of
`cardiac growth. Ang II functions as a pressor hormone and is
`involved the pathophysiology of several forms of hyperten
`S1O.
`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 of the
`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 vasopressin peptide from the pituitary
`gland and, on the other hand, of aldosterone from the adrenal
`glomerulosa. 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 of the
`target cell. It has been possible to identify receptor subtypes
`that are termed, e.g., AT1 - and AT 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
`sives or for the treatment of congestive heart failure, among
`otherindications. Ang II 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 lower blood 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 of ACE inhibitors involves
`prevention of formation of the vasoconstrictorpeptide 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
`stance P. Prevention of the degradation of bradykinin 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
`
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`40
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`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1014, p. 003
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`

`

`US 7,468.390 B2
`
`3
`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 of the 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)-N-(1-car
`boxy-2-methyl-prop-1-yl)-N-pentanoyl-N-2:(1H-tetrazol
`5-yl)biphenyl-4-yl-methylamine of formula (I)
`
`O
`
`CH3
`
`CH
`1.
`Neil
`
`-CH3
`CH3N,
`IC1
`y
`C
`C
`H
`1.
`Nc6, N N 1. NY
`
`4
`R is hydroxy, alkoxy of 1 to 7 carbons, or NH;
`n 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
`R is benzyl:
`R is hydrogen;
`n is an integer from 1 to 9; and
`R 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
`R is benzyl:
`R is hydrogen;
`n is one; and
`R is hydroxy.
`The preparation of the selective NEP inhibitors of formula
`(II), wherein R is other than trifluoromethyl are disclosed by
`Delaney et al. in U.S. Pat. No. 4,722,810. The preparation of
`the selective NEP inhibitors of formula (II), wherein R is
`trifluoromethyl are disclosed by Delaney et al. in U.S. Pat.
`No. 5,223,516.
`NEP inhibitors within the scope of the present invention
`include compounds disclosed in U.S. Pat. No. 4,610,816,
`herein incorporated by reference, including in particular
`N-N-1 (S)-carboxyl-3-phenylproplyl
`(S)-phenylalanyl-(S)-isoserine and N-N-(1S)-carboxy-2-
`phenyl)ethyl-(S)-phenylalanyl-B-alanine; compounds dis
`closed in U.S. Pat. No. 4,929,641, in particular, N-2(S)-
`mercaptomethyl-3-(2-methylphenyl)-propionylmethionine;
`SQ 28.603 (N-2-(mercaptomethyl)-1-oxo-3-phenylpropyl
`B-alanine), disclosed in South African Patent Application No.
`84/0670: UK 69578 (cis-4-1-2-carboxy-3-(2-methoxy
`ethoxy)propyl-cyclopentylcarbonylaminol-cyclohexan
`ecarboxylic 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., com
`pounds 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 U.S. 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-
`methoxyethoxy)propyl-1-cyclopentanecarboxamido-1-cy
`clohexanecarboxylic acid; the compounds disclosed in GB
`022 18983, particularly 3-(1-6-endo-hydroxymethylbicyclo
`2.2.1]heptane-2-exo-carbamoylcyclopentyl)-2-(2-meth
`oxyethyl)propanoic acid; the compounds disclosed in WO
`92/14706, particularly N-(1-(3-(N-t-butoxycarbonyl-(S)-
`prolylamino)-2(S)-t-butoxy-carbonylpropyl)cyclopentan
`ecarbonyl)-O-benzyl-(S)-serine methyl ester; the compounds
`disclosed in EP 00343911; the compounds disclosed in JP
`06234754; the compounds disclosed in EP 00361365, par
`ticularly
`4-2-(Mercaptomethyl)-1-oxo-3-phenylpropyl
`aminobenzoic acid; the compounds disclosed in WO
`90/09374, particularly 3-1-(Cis-4-carboxycarbonyl-cis-3-
`butylcyclohexyl-r-1-carboarnoyl)cyclopentyl-2S-(2-meth
`oxyethoxymethyl)propanoic acid; the compounds disclosed
`
`10
`
`15
`
`25
`
`35
`
`40
`
`45
`
`(I)
`
`30
`
`O
`
`OH
`
`HN1SN
`V
`/
`NFN
`
`and is disclosed in EP 0443983 A and U.S. Pat. No. 5,399,
`578, the disclosures of which are incorporated herein in their
`entirety as if set forth herein.
`ANEP inhibitor useful in said combination is a compound
`of the formula (II)
`
`50
`
`R
`
`O
`
`R3
`
`O
`
`(II)
`
`55
`
`HS-CH-CH-C-NH-CH-(CH)-C-R
`
`and pharmaceutically acceptable salts thereof,
`wherein
`R is alkyl of 1 to 7 carbons, trifluoromethyl, phenyl, sub
`stituted phenyl, -(CH2) a-phenyl, or —(CH)
`4-Substituted phenyl,
`R is hydrogen, alkyl of 1 to 7 carbons, phenyl, Substituted
`phenyl, -(CH2)
`-phenyl, or —(CH) a-substi
`tuted phenyl:
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`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1014, p. 004
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`

`

`5
`in JP 07157459, particularly N-((2S)-2-(4-biphenylmethyl)-
`4-carboxy-5-phenoxyvaleryl)glycine; the compounds dis
`closed in WO 94/15908 particularly N-(1-(N-hydroxycar
`bamoylmethyl)-1-cyclopentanecarbonyl)-L-phenylalanine;
`the compounds disclosed in U.S. Pat. No. 5.273,990 particu
`larly (S)-(2-biphenyl-4-yl)-1-(1H-tetrazol-5-yl)ethylamino)
`methylphosphonic 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 U.S. 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, N-N-I(L)-1-(2,2-dim
`ethyl-1
`3-dioxolan-4-yl)-methoxycarbonyl-2-
`phenylethyl-L-phenylalanyl-(R)-alanine, N -N-I(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,
`N-(S)-3-mercapto-2-(2-meth
`ylphenyl)propionyl-(S)-2-methoxy-(R)-alanine, N-1-1
`(S)-benzyloxycarbonyl-3-phenylpropylaminocyclopentyl
`carbonyl-(S)-isoserine,
`N-1-1 (S)-carbonyl-3-
`phenylpropylaminol-cyclopentylcarbonyl-(S)-isoserine,
`1,1-dithiobis-2(S)-(2-methylbenzyl)-1-oxo-3,1-propaned
`lyl-bis-(S)-isoserine, 1,1'-dithiobis-2(S)-(2-methylben
`Zyl)-1-oxo-3,1-propanediyl-bis-(S)-methionine, N-(3-phe
`nyl-2-(mercaptomethyl)-propionyl)-(S)-4-
`(methylmercapto)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)-cyclopentane
`carbonyl-(S)-isoserine, N-1-(acetylthiomethyl)cyclopen
`tane-carbonyl-(S)-methionine
`ethyl
`ester,
`3(S)-2-
`(acetylthiomethyl)-3-phenyl-propionylamimo-e-caprolac
`tam; and the compounds disclosed in WO 93/10773
`particularly N-(2-acetylthiomethyl-3-(2-methylphenyl)pro
`pionyl)-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.
`55
`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 U.S.
`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 ethylether and 0.113 mL (0.848
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`mmol) of triethanolamine in 1 mL of ethyl acetate. The solid
`was collected and dried melting at 69-71°C.
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`HO
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`NH2
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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 of ethyl 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 114-115° 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 of the 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 pharmaceutically
`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.
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`BIOCON PHARMA LTD (IPR2020-01263) Ex. 1014, p. 005
`
`

`

`7
`It can be shown that combination therapy with Valsartan
`and a NEP inhibitor results in a more effective anti-hyperten
`sive therapy (whetherformalignant, essential, reno-vascular,
`diabetic, isolated Systolic or other secondary type of hyper
`tension) through improved efficacy, 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 insuffi
`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
`roderma, 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 ratchow
`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.
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`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
`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
`their juncture 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 prior to experimentation, animals are
`anesthetized with isoflurane and catheters are implanted in
`the femoral artery and vein for measuring arterial pressure,
`collection of blood and administration of test compounds.
`Rats are allowed to recover for 48 hours while tethered in a
`Plexiglas home cage, which also serves as the experimental
`chamber.
`Chronic Study Procedure:
`This procedure is the same as above except that rats are
`implanted with a radiotransmitter, 7-10 days prior to the
`unilateral nephrectomy and initiation of DOCA and salt. In
`addition, rats do not undergo Surgery for placement of femo
`ral arterial and venous catheters. Radiotransmitters are
`implanted as described in Bazilet al., “Telemetric Monitoring
`of Cardiovascular Parameters in Conscious Spontaneously
`Hypertensive Rats'. J. Cardiovasc. Pharmacol., Vol. 22, pp.
`897-905 (1993).
`Protocols are then set-up on the computer for measurement
`of blood pressure, heart rate, etc., at pre-determined time
`points. Baseline data is collected at various time points and
`over various time intervals. For example, baseline or pre-dose
`values usually consist of data collection and averaging over
`three consecutive, 24-hour time periods prior to drug admin
`istration.
`Blood pressure, heart rate and activity are determined at
`various pre-selected time points before, during, and after drug
`administration. All measurements are performed in unre
`strained and undisturbed animals. The maximum study time,
`determined by battery life, could be as long as nine months.
`For studies of this duration, rats are dosed orally (1-3 mL/kg
`vehicle), no more than twice daily or drug is administered via
`the drinking water or mixed with food. For studies of a shorter
`duration, that is, up to 8 weeks, drugs are given via s.c.
`implanted osmotic minipumps. Osmotic minipumps are
`selected based on drug delivery rate and time. Valsartan dos
`ages range from 1-10 mg/kg/day and N-(3-carboxy-1-oxo
`propyl)-(4S)-(p-phenylphenylmethyl)-4-amino