Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 1 of 15
`
`Kowa Company, Ltd. et al. v. Aurobindo Pharma Limited et al.,
`Civil Action No. 14-CV-2497 (PAC) (and related cases)
`
`
`
`Exhibit 9 to Supplemental Declaration of
`Thomas R. Burns, dated June 10, 2015, in
`support of Defendants’ Joint Responsive
`Claim Construction Brief
`
`
`
`

`

`Case 1:14-cv-02758-pAc DocunlllllllllllllllllllllllllllllllllllllfllllllllllllllllUIIlllllllllllalIIl
`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 2 of 15
`U8005827871A
`
`[19]
`United States Patent
`5,827,871
`[11] Patent Number:
`King et al. Oct. 27, 1998
`[45] Date of Patent:
`
`
`
`An‘omey, Agent, or Firm—Nora Stein-Fernandez; Janice E.
`Williams
`
`[57]
`
`ABSTRACT
`
`Use of a compound of general formula (I):
`
`R1
`
`NRZR3
`
`Formula (I)
`
`NH
`
`wherein
`
`R1 represents hydrogen, halogen, trifluoromethyl, nitro,
`hydroxy, C1_5alkyl, C1_6alkoxy, arle1_6alkoxy,
`7C02R4, 7(CH2),,CN, %CH2),,CONR5R6, 7(CH2)
`"SOZNRSRO, C1_6alkanoylamino(CH2)n, or
`CM)alkylsulphonylamino(CH2)n;
`R4 represents hydrogen, C1_fialkyl or arle1_5alkyl;
`R5 and R6 each independently represent hydrogen or
`Cmalkyl, or R5 and R6 together with the nitrogen atom
`to which they are attached form a ring;
`n represents 0,
`l or 2; and
`R2 and R3 each independently represent hydrogen,
`Cl_6alkyl or benzyl or together with the nitrogen atom
`to which they are attached form a pyrrolidino, piperi-
`dino or hexahydroazepino ring;
`or a physiologically acceptable salt thereof, in the manufac-
`ture of a medicament for the treatment of a condition where
`a 5-HT1-like agonist is indicated, for example migraine.
`Novel compounds of formula (I), processes for preparing
`them and pharmaceutical compositions containing them are
`also described.
`
`17 Claims, No Drawings
`
`[54] MEDICAMENTS 1,2,3,4—
`TETRAHYDROCARBAZOLES AND 5-HT1
`AGONIST USE THEREOF
`
`[75]
`
`Inventors: Francis David King; Laramie Mary
`Caster, both of Harlow; Alberto Julio
`Kaumann, Trumpington; Rodney
`Christopher Young, Hertford, all of
`England
`
`73] Assignee: Smithkline Beecham plc, England
`
`
`
`21] Appl. No.: 442,720
`
`22]
`
`Filed:
`
`May 17, 1995
`
`Related US. Application Data
`
`63]
`
`30]
`
`Continuation—impart of Ser. No. 167,846, filed as PCT/
`GB92/01082, Jun. 17, 1992, Pat. No. 5,464,864.
`
`Foreign Application Priority Data
`
`Jun. 26, 1991
`
`[GB]
`
`United Kingdom ................. .. 9113802
`
`Int. Cl.6 ....................... .. A61K 31/40; C07D 209/82
`51]
`52] US. Cl.
`........................ .. 514/411; 514/212; 514/323;
`540/602; 546/200; 548/448; 548/449
`58] Field of Search ................................... .. 514/411, 212,
`514/323; 540/602; 546/200; 548/448, 440,
`449
`
`56]
`
`References Cited
`FOREIGN PATENT DOCUMENTS
`
`0 242 518 10/1997 European Pat. Off. .
`
`Primary Examiner—Jacqueline IIaley
`
`MYLAN(Pitav) 037192
`
`
`

`

`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 3 of 15
`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 3 of 15
`
`1
`
`5,827,871
`
`MEDICAMENTS 1,2,3,4-
`TETRAHYDROCARBAZOLES AND 5-HT1
`AGONIST USE THEREOF
`
`This is a continuation-in-part application of application
`Ser. No. 08/167,846, filed Dec. 23, 1993, now US. Pat. No.
`5,464,864, which is a U.S. national phase application of
`PCT/GB92/01082, filed Jun. 17, I992, claiming priority
`from GB 9113802, filed Jun. 26, 1991.
`The present invention relates to certain tetrahydrocarba-
`zole derivatives for use in the treatment of disorders char-
`
`acterised by excessive vasodilatation, in particular the treat-
`ment of migraine.
`Migraine is a non-lethal disease which has been reported
`to be suffered by one in ten individuals. The main symptom
`is headache; other symptoms include vomiting and photo-
`phobia. Currently,
`the most widely used treatment
`for
`migraine involves administration of ergotamine, dihydroer-
`gotamine or methysergide, which are also used prophylac-
`tically. These drugs are inter alia agonists of 5HTl-like
`receptors but also have other actions; treatment with them is
`associated with a number of adverse side effects. In addition,
`some patients experience a “withdrawal headache” follow-
`ing the cessation of treatment with an ergot product, such as
`ergotamine, causing them to repeat the treatment and result-
`ing in a form of addiction. More recently various tryptamine
`derivatives have been proposed for potential use in treating
`migraine.
`In View of the foregoing, there is clearly a need for the
`provision of effective and safe medicaments for the treat-
`ment of migraine.
`US. Pat. Nos. 4,257,952, 4,172,834, 4,062,864 and
`3,959,309 disclose a broad class of tetrahydrocarbazoles of
`the formula:
`
`Q7
`
`Q3
`
`Qi
`
`Q4
`
`N
`IL
`
`N=B
`
`wherein N =B is inter alia —NHR‘ or —NR'R" where R‘ and
`R" are lower alkyl, aryl-lower alkyl or together form a
`heterocyclic ring; R is inter alia hydrogen; O1 is inter alia
`hydrogen, halogen,
`lower alkoxy, cyano, —C02R1 or
`—CONR2R3 (where R1 may be hydrogen, lower alkyl or
`—CH2A1' and R2 and R3 are hydrogen,
`lower alkyl or
`together form a heterocyclic ring); Q2 is inter alia hydrogen,
`aryl-(lower alkoxy), hydroxy,
`trihalomethyl, nitro or
`alkanoylamino, and Q3 and Q4 may each be inter alia
`hydrogen. These compounds are said to have analgetic,
`psychotropic and anthistaminic activities.
`It has now surprisingly been found that certain tetrahy-
`drocarbazoles are agonists and partial agonists at SIITl-like
`receptors and are expected to have utility in the treatment of
`conditions wherein a 5—HTl—like agonist or partial agonist is
`indicated, in particular conditions associated with cephalic
`pain such as migraine, cluster headache and headache asso-
`ciated with vascular disorders. In this specification the term
`‘5-HT1-like agonist’ will hereinafter be used to include
`partial agonists at this receptor.
`The present invention therefore provides the use of com—
`pounds of general formula (I):
`
`5
`
`10
`
`20
`
`30
`
`40
`
`50
`
`60
`
`V
`“R21”
`
`“mm (1)
`
`2
`
`l
`
`NH
`
`1
`
`R
`
`wherein:
`
`R1 represents hydrogen, halogen, trifluoromethyl, nitro,
`hydroxy, C1_6alkyl, Cl_6alkoxy, arlel_6alkoxy,
`—CO2R4, —(CH62),,CN, —(CH2)”CONR5R6,
`—(CH2),,SOZNR5R , C1,6alkanoylamino(CH2)n, or
`C1_6alkylsulphonylamino(CH2)n;
`R4 represents hydrogen, C1_6alkyl or arle1_6alky'l;
`Rs and R6 each independently represent hydrogen or
`C1_6alkyl, or R5 and R6 together with the nitrogen atom
`to which they are attached form a ring;
`n represents 0, 1 or 2; and
`R2 and R3 each independently represent hydrogen,
`Cl_6alkyl or benzyl or together with the nitrogen atom
`to which they are attached form a pyrrolidino, piperi-
`dino or hexahydroazepino ring;
`and physiologically acceptable salts thereof, in the manu-
`facture of a medicament for the treatment of a condition
`where a 5-HT1-like agonist is indicated, in particular the
`treatment or prophylaxis of migraine.
`The invention also provides a method of treatment of a
`condition wherein a 5—HT1—like agonist
`is indicated,
`in
`particular migraine, which comprises administering to a
`subject in need thereof an eifective amount of a compound
`of formula (I) or a physiologically acceptable salt thereof.
`Suitably R1 represents hydrogen, halogen, cyano,
`hydroxy, C1_Galkoxy, arle1_Galkoxy, —C02R4,
`—(CHZ),,CONRSR6 or —(CH2),,SOZNR5R6; and R2 and R3
`each independently represent hydrogen or CLsalkyl.
`It will be appreciated that compounds of formula (I) may
`contain one or more assymetric centres, and such coni-
`pounds will exist as optical
`isomers (enantiomers). The
`invention thus includes all such enantiomers and mixtures,
`including racemic mixtures, thereof.
`In the compounds of formula (I) a halogen atom may be
`a fluorine, chlorine, bromine or iodine atom. An alkyl group
`or moiety may have a straight or branched chain. Suitable
`aryl groups include for example unsaturated monocyclic or
`bicyclic rings and partially saturated bicyclic rings of up to
`12 carbon atoms, such as phenyl, naphthyl and tetrahy-
`dronaphthyl. When R5 and R6 together with the nitrogen
`atom form a ring, this is preferably a 5 to 7—membered
`saturated heterocyclic ring, which may optionally contain a
`further heteroatom selected from oxygen, sulphur or nitro-
`gen. Suitable rings thus include pyrrolidino, piperidino,
`piperazino and morpholino.
`In the above compounds R1 preferably represents halogen
`(e.g. bromine), CF3, C1_6alkoxy (e.g. methoxy), (CH2)nCN,
`—(CH2)nCONR5R6, —(CH2)nSOZNR5R6 or
`C1_6alkanoylamino. Most preferably Rl represents a group
`—(CH2)nCONR5RG wherein n represents 0 and RS and RG
`each independently represent hydrogen, methyl, ethyl or
`propyl. Advantageously, R5 and R6 independently represent
`hydrogen or methyl.
`When R1 represents —C02R4, then R4 preferably repre-
`sents C1_6alkyl.
`R2 and R3 each preferably represent hydrogen, methyl or
`ethyl. Most preferably NRZR3 is —NH2.
`For use according to the present invention the compound
`of formula (I) is preferably a partial agonist.
`Suitable physiologically acceptable salts will be apparent
`to those skilled in the art and include for example acid
`
`MYLAN(Pitav) 037193
`
`
`

`

`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 4 of 15
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`5,827,871
`
`4
`3-i-propy1amino-6-carboxamido-1,2,3,4-
`tetrahydrocarbazole oxalate,
`3-dimethylamino-6-carboxamido-l ,2,3,4-
`tetrahydrocarbazole oxalate,
`3-benzylamino-6-carboxamido-1,2,3,4-tetrahydrocarbazole
`oxalate,
`3—pyrrolidinyl—6—carboxamido—1,2,3,4—tetrahydrocarbazole
`oxalate, and
`3-(N-(methyl)ethylamino)-6-earboxamido- l,2,3,4-
`tetrahydrocarbazole oxalate,
`3-amino-6-(2-carboxamidoethyl)-1,2,3,4-
`tetrahydrocarbazole oxalate.
`In a further aspect the present invention provides a novel
`compound of formula (I) e,g. a compound of formula (IA)
`or any of the above-named compounds (in free base form or
`as a physiologically acceptable salt) for use as a therapeutic
`agent, in particular as a 5-HT1-like agonist or partial agonist,
`for example for the treatment of migraine.
`The invention also provides a process for the preparation
`of novel compounds of formula (I).
`Compounds of formula (I) may be prepared by methods
`known in the art for the preparation of tetrahydrocarbazoles,
`for example:
`A) Reaction of a compound of formula (II):
`Rl
`
`Formula (II)
`
`10
`
`20
`
`: NHNH;
`
`30
`
`(wherein R1 is as hereinbefore defined) or an acid addition
`salt thereof with a compound of formula (III):
`
`::
`
`NRZR3
`
`0
`
`Formula (III)
`
`(wherein R2 and R3 are as hereinbefore defined) or an
`N-protected derivative thereof; or
`B) Reaction of a compound of formula (IV):
`
`40
`
`R1
`
`Z
`
`Formula (IV)
`
`NH
`
`(wherein R1 is as defined for formula (I) and Z is a leaving
`group) with a compound of formula HNR2R3;
`C) Reacting a compound of formula (V):
`
`H2N(CHZ)
`
`50
`
`NRZR3
`
`Formula (V)
`
`NH
`
`60
`
`with an acylating or sulphonylating agent;
`D) Conversion of one compound of formula (I) into
`another compound of formula (I) eg.
`(i) to prepare a compound of formula (I) wherein R1
`represents —(CH2),,CONH2 or C02R4, hydrolysis of a
`compound of formula
`wherein R1
`represents
`—(CIIz)nCN, or an N-protected derivative thereof;
`(ii) to prepare a compound of formula (I) wherein R1
`represents —CONR5R6, amination of a compound of for-
`mula (I) wherein R1 represents —C02H, or an N-protected
`derivative thereof; or
`(iii) to prepare a compound of formula (I) wherein one of
`R2 and R3 is hydrogen and the other is C1_6alkyl, alkylation
`of a compound (I) in which R2 and R3 are both hydrogen;
`
`3
`addition salts such as those formed with inorganic acids e.g.
`hydrochloric, sulphuric or phosphoric acids and organic
`acids e.g. succinic, maleic, acetic or fumaric acid. Other
`non-physiologically acceptable salts e.g. oxalates may be
`used for example in the isolation of compounds of formula
`(I), and are included within the scope of this invention. Also
`included within the scope of the invention are solvates and
`hydrates of compounds of formula
`It is believed that compounds of formula (I) wherein R2
`and R3 both represent hydrogen are novel. Thus in a further
`aspect the present invention provides compounds of formula
`(IA):
`
`Rl
`
`NH;
`
`Formula (IA)
`
`NH
`
`wherein R1 is as hereinbefore defined, and salts thereof.
`The present
`invention further provides the following
`specific compounds which are also believed to be novel:
`3-Amino-6-cyano-1,2,3,4-tetrahydrocarbazole
`hydrochloride,
`(+)-3-amino-6-carboxamido-1,2,3,4-tetrahydrocarbazole
`hydrochloride,
`(—)-3-amino-6-carboxamido-1,2,3,4-tetrahydrocarbazole
`hydrochloride,
`3—amino—6—methoxy—1,2,3,4—tetrahydrocarbazole
`hydrochloride,
`3-amino-6-bromo-1 ,2,3,4-tetrahydrocarbazole
`hydrochloride,
`3-amino-6-methyl-1,2,3,4-tetrahydrocarbazole oxalate,
`3-amino-6-ethoxycarbonyl- l ,2,3,4-tetrahydrocarbazole
`oxalate,
`3-amino-6-(N-methyl carboxamido)-1,2,3,4-
`tetrahydrocarbazole hemioxalate,
`3-amino-6-cyanomethyl-1 ,2,3,4-tetrahydrocarbazole
`oxalate,
`3-amino-6-(N-methylsulphonamidomethyl)-1,2,3,4-
`tetrahydrocarbazole oxalate,
`3—amino—6—chloro—1,2,3,4—tetrahydrocarbazole oxalate,
`3-amino-6-trifluoromethyl-1,2,3,4-tetrahydrocarbazole
`oxalate,
`3-amino-6-n-butyloxy-1,2,3,4-tetrahydrocarbazole oxalate,
`3-amino-6-sulph on amido-l ,2,3,4-tetrahydrocarb azole
`oxalate,
`3-amino-6-nitro-1,2,3,4-tetrahydrocarbazole oxalate,
`3-amino-6-(N,N-dimethylcarboxamido)- 1,2,3,4-
`tetrahydrocarbazole hemioxalate,
`3-amino-6-(piperidin-1-ylcarbonyl)-1,2,3,4-
`tetrahydrocarbazole hydrochloride,
`3-amino-6-(pyrrolidin-1 -ylcarbonyl)-1,2,3,4-
`tetrahydrocarbazole hydrochloride,
`3—amino—6—(N,N—diethylearboxamido)—1 ,2,3,4—
`tetrahydrocarbazole hydrochloride,
`3-Amino-6-(acetamido)-1,2,3,4-tetrahydrocarbazole
`oxalate,
`3-amino-6-methanesulphonamido-1,2,3,4-
`tetrahydrocarbazole oxalate,
`3-amino-6-carboxamidomethyl-1,2,3,4-tetrahydrocarbazole
`hydrochloride,
`3-niethylamino-6-carboxamido-1,2,3,4-tetrahydrocarbazole
`oxalate,
`3-ethylamino-6-carboxamido-1,2,3,4-tetrahydrocarbazole
`oxalate,
`3—n—propylamino—6—carboxamido—1,2,3,4—
`tetrahydrocarbazole oxalate,
`
`MYLAN(Pitav) 037194
`
`
`

`

`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 5 of 15
`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 5 of 15
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`5,827,871
`
`5
`
`(iv) to prepare a compound of formula (I) wherein R1
`represents hydroxy, cleavage of a compound wherein R1
`represents alkoxy or aralkoxy;
`followed if necessary by deprotection of any protected
`nitrogen atoms and if desired by salt formation.
`Process (A), which is a form of the Fischer indole
`synthesis, may be carried out using methods well known in
`the art. Thus, the reaction may be effected in a solvent, for
`example an alcohol such as ethanol or butanol; or acetic
`acid, and at a temperature in the range 0° to 150° C.
`Hydrazines of formula (II), which are usually employed
`as the hydrochloride salt, are known compounds, or may be
`prepared by conventional methods.
`A cyclohexanone of formula (III) may be prepared by
`oxidation of the corresponding cyclic alcohol, using an
`oxidising agent such as pyridinium chlorochromate, pyri-
`dinium dichromate, dipyridine Cr
`(VI) oxide, sodium
`hypochlorite, calcium hypochlorite or manganese dioxide.
`The leaving group Z in the compounds of formula (IV)
`may be for example a halogen atom, or a sulphonyloxy
`group eg. p-toluenesulphonyloxy or methanesulphonyloxy.
`Process (B) may be effected in an inert organic solvent, such
`as an alcohol eg. methanol or an ether eg. tetrahydrofuran
`and at a temperature in the range 0° to 150° C. Compounds
`of formula (IV) may be obtained by reacting a hydrazine of
`formula (II) with an appropriately substituted cyclohex-
`anone compound. When Z is acyloxy or sulphonyloxy this
`may be prepared from a compound (IV) wherein Z is
`hydroxy, using standard procedures. Suitable acylating and
`sulphonylating agents which may be used in process (C)
`include carboxylie and suilphonic acid chlorides (e.g. acetyl
`chloride or methanesulphonylchloride) alkyl esters, acti-
`vated esters and symmetrical and mixed anhydrides. The
`reaction may be carried out in an organic solvent such as a
`haloalkane (e.g. dichloromethane), an amide (e.g. N,N-
`dimethylformamide; an ether (e.g.
`tetrahydrofuran) or a
`tertiary amine such as pyridine. In general a base will also
`be used, e.g. triethylamine, dimethylaminopyridine, or an
`alkali metal carbonate or bicarbonate. The reaction may be
`effected at a temperature in the range of —10° to 100° C.
`Compounds of formula (V) may be prepared by methods
`analogous to processes (A) and (B) hereinbefore described.
`Alternatively a compound of formula (V) may be obtained
`by subjecting a compound of formula (I) wherein R1 is nitro
`to reduction, e.g. by catalytic hydrogenation.
`It is well known in the chemical art that hydrolysis of a
`nitrile initially results in an amide, which can be further
`hydrolysed to an acid. It will therefore be appreciated that
`the precise product of process (Di) will depend upon the
`reaction conditions chosen for the hydrolysis. To obtain a
`compound wherein R1 represents H2NCO— the hydrolysis
`is preferably effected using hydrogen peroxide in the pres-
`ence of an alkali hydroxide e.g. sodium hydroxide,
`in a
`solvent such as an alcohol e.g. methanol. Other suitable
`means of hydrolysis include acetic acid and BF3; or formic
`acid and hydrobromic or hydrochloric acid. To prepare a
`compound wherein R1 represents —C0()H acid or base
`catalysed hydrolysis may be used.
`Process (Dii) may be effected by reacting a compound of
`formula (I) wherein R1 is —C02H with an amine HNRSRS,
`in the presence of a coupling agent e.g. dicyclohexylcarbo-
`diimide or N,N'-carbonyldiimidazole. Alternatively the car-
`boxylie acid starting material may first be reacted to form an
`activated derivative of the carboxyl group, for example an
`acid chloride, acid anhydride or activated ester, which is
`then reacted directly with an amine HNRSRS. The carboxylic
`acid may also be activated in situ for example by treating
`with hexamethylphosphoroustriamide.
`
`10
`
`20
`
`30
`
`40
`
`50
`
`60
`
`6
`Alkylation according to process (Diii) may be effected by
`reacting an amine of formula (I) with an acylating agent, for
`example an anhydride, such as acetic or propionic
`anhydride, to form an intermediate in which one of R2 or R3
`is —C(O)C1_6alkyl, followed by reduction of said interme-
`diate to give the desired product. Other reagents and con-
`ditions will be apparent to those skilled in the art.
`Cleavage according to process (Div) may be effected by
`reduction, using methods well known in the art.
`It will be appreciated that in many of the above reactions
`it will be necessary to protect the group —NR2R3 when one
`or both of the groups R2 and R3 represent hydrogen. Suitable
`N—protecting groups are well—known in the art and include
`for example acyl groups such as acetyl,
`trifluoroacetyl,
`benzoyl, methoxycarbonyl,
`t-butoxycarbonyl, benzyloxy-
`carbonyl or phthaloyl; and aralkyl groups such as benzyl,
`diphenylniethyl or triphenylmethyl. When R2 and R3 both
`represent hydrogen the nitrogen atom is preferably protected
`as the phthalimide. The protecting groups should be easily
`removable at
`the end of
`the reaction sequence.
`N-deprotection may be effected by conventional methods,
`for example a phthaloyl group may be removed by reaction
`with hydrazine; an acyl group such as benzoyl may be
`cleaved by hydrolysis and an aralkyl group such as benzyl
`may be cleaved by hydrogenolysis.
`When a compound of formula (I) is obtained as a mixture
`of enantiomers these may be separated by conventional
`methods, for example by reaction of the mixture with a
`suitable optically active acid such as d—tartaric acid, l—malic
`acid, 1-mandelic acid,
`l-gulonic acid or 2,3:4,6-di-O-
`isopropylidene-keto-L-gulonic acid to give two diastereoi-
`someric salts which may be separated eg. by crystallisation.
`Alternatively mixtures of enantiomers may be separated by
`chromatography, for example on a chiral HPLC column.
`Compounds of formula (I) have been found to be agonists
`and partial agonists at 5HT1-1ike receptors and are expected
`to have utility in the treatment and/or prophylaxis of
`migraine, and other conditions associated with cephalic
`pain.
`For use in medicine, the compounds of the present inven-
`tion are usually administered as a standard pharmaceutical
`composition. The present invention therefore provides in a
`further aspect pharmaceutical compositions comprising a
`novel compound of formula (I) or a physiologically accept-
`able salt thereof and a physiologically acceptable carrier.
`The compounds of formula (I) may be administered by
`any convenient method, for example by oral, parenteral,
`buccal, sublingual, nasal, rectal or transdermal administra-
`tion and the pharmaceutical compositions adapted accord-
`ingly.
`The compounds of formula (I) and their physiologically
`acceptable salts which are active when given orally can be
`formulated as liquids, for example syrups, suspensions or
`emulsions, tablets, capsules and lozenges.
`A liquid formulation will generally consist of a suspen-
`sion or solution of the compound or physiologically accept-
`able salt
`in a suitable liquid carrier(s) for example an
`aqueous solvent such as water, ethanol or glycerine, or a
`non-aqueous solvent, such as polyethylene glycol or an oil.
`The formulation may also contain a suspending agent,
`preservative, flavouring or colouring agent.
`A composition in the form of a tablet can be prepared
`using any suitable pharmaceutical carrier(s) routinely used
`for preparing solid formulations. Examples of such carriers
`include magnesium stearate, starch,
`lactose, sucrose and
`cellulose.
`A composition in the form of a capsule can be prepared
`using routine encapsulation procedures. For example, pellets
`
`MYLAN(Pitav) 037195
`
`
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`5,827,871
`
`7
`containing the active ingredient can be prepared using
`standard carriers and then filled into a hard gelatin capsule;
`alternatively, a dispersion or suspension can be prepared
`using any suitable pharmaceutical carrier(s), for example
`aqueous gums, celluloses, silicates or oils and the dispersion
`or suspension then filled into a soft gelatin capsule.
`Typical parenteral compositions consist of a solution or
`suspension of the compound or physiologically acceptable
`salt in a sterile aqueous carrier or parenterally acceptable oil,
`for example polyethylene glycol, polyvinyl pyrrolidone,
`lecithin, arachis oil or sesame oil. Alternatively, the solution
`can be lyophilised and then reconstituted with a suitable
`solvent just prior to administration.
`Compositions for nasal administration may conveniently
`be formulated as aerosols, drops, gels and powders. Aerosol
`formulations typically comprise a solution or fine suspen-
`sion of the active substance in a physiologically acceptable
`aqueous or non-aqueous solvent and are usually presented in
`single or mnltidose quantities in sterile form in a sealed
`container, which can take the form of a cartridge or refill for
`use with an atomising device. Alternatively the sealed con-
`tainer may be a unitary dispensing device such as a single
`dose nasal
`inhaler or an aerosol dispenser fitted with a
`metering valve which is intended for disposal once the
`contents of the container have been exhausted. Where the
`dosage form comprises an aerosol dispenser, it will contain
`a propellant which can be a compressed gas such as com—
`pressed air or an organic propellant such as a fluorochloro—
`hydrocarbon. The aerosol dosage forms can also take the
`form of a pump-atomiser.
`Compositions suitable for buccal or sublingual adminis-
`tration include tablets, lozenges and pastilles, wherein the
`active ingredient is formulated with a carrier such as sugar
`and acacia, tragacanth, or gelatin and glycerin.
`Compositions for rectal administration are conveniently
`in the form of suppositories containing a conventional
`suppository base such as cocoa butter.
`Compositions suitable for transdermal administration
`include ointments, gels and patches.
`Preferably the composition is in unit dose form such as a
`tablet, capsule or ampoule.
`Each dosage unit for oral administration contains prefer-
`ably from 1 to 250 mg (and for parenteral administration
`contains preferably from 0.1 to 25 mg) of a compound of the
`formula (I) or a physiologically acceptable salt
`thereof
`calculated as the free base.
`The physiologically acceptable compounds of the inven-
`tion will normally be administered in a daily dosage regimen
`(for an adult patient) of, for example, an oral dose of
`between 1 mg and 500 mg, preferably between 10 mg and
`400 mg, e.g. between 10 and 250 mg or an intravenous,
`subcutaneous, or intramuscular dose of between 0.1 mg and
`100 mg, preferably between 0.1 mg and 50 mg, e.g. between
`1 and 25 mg of the compound of the formula (I) or a
`physiologically acceptable salt thereof calculated as the free
`base, the compound being administered 1 to 4 times per day.
`Suitably the compounds will be administered for a period of
`continuous therapy, for example for a week or more.
`BIOLOGICAL DATA
`
`5—HT1—like Receptor Screen
`Dog Saphcnous Vein
`IIelicoids of dog saphenous vein were set up at 37° C. in
`modified Krebs solution at a resting force of 10 mN. The
`solution also contained 1 ,umol/l each of ketanserin prazosin,
`atropine and mepyramine, 6 ymol/l cocaine and 200 ,umol/l
`ascorbate. Nearly isomeric contractions were measured with
`
`8
`force transducers on a polygraph. The tissues were exposed
`twice to 5—hydroxytryptamine (5—HT) 2 ,umol/l followed by
`washes. A cumulative concentration-effect curve was
`
`determined, followed by a curve to 5-HT in the presence of
`the highest used concentration of test compound. Contrac-
`tions caused by the test compound were compared with
`those caused by 5-HT. The intrinsic activity of the test
`compound was calculated as the ratio of the maximum test
`compound-induced effect over the effect caused by 2 Mmol/l
`5-HT. The ECSO of the test compound was estimated from
`the corresponding effect curve. When appropriate equilib-
`rium dissociation constants Kp were estimated by the
`method of Marano & Kaumann (1976, J. Pharmacol. Exp.
`Ther. 198, 5187525).
`In this screen the compounds of Examples 2, 4, 5, 6, 9, 10,
`11, 13, 17, 18, 21 and 24 had ECSO’S in the range 0.1 to 15
`ymol.
`RABBIT BASILAR ARTERY
`METHODS
`
`Experiments were performed in intracranial arteries from
`rabbit isolated basilar artery in a similar method to one
`described previously (Parsons and Whalley, 1989. Eur J
`Pharmacol 174, 189—196.).
`In brief, rabbits were killed by overdose with anaesthetic
`(sodium pentobarbitone). The whole brain was quickly
`removed and immersed in ice cold modified Krebs solution
`and the basilar artery removed with the aid of a dissecting
`microscope. The Krebs solution was of the following com-
`position (mM) Na+ (120); K+ (5); Ca2+ (2.25); Mg2+ (0.5);
`CI" (98.5); SO42" (1); ED'I'A (0.04), equilibrated with 95%
`025% C02. The endothelium was removed by a gentle
`rubbing of the lumen with a fine metal wire. Arteries were
`then cut into ring segments (ca 4—5 mm wide) and set up for
`recording of isometric tension in 50 m1 tissue baths in
`modified Krebs solution with the additional supplement of
`(111M); Na2+ (20); fumarate 10); pyruvate (5); L-glutamate
`(5) and glucose (10). The arteries were then placed under a
`resting force of 3—4 mN maintained at 37° C. and the
`solution bubbled with 95% 02/5% C02.
`After tests for initial reactivity with 90 mM KCl depola-
`rising solution and for lack of aeetyleholine-indueed relax-
`ation of 5-HT (10 mM) precontraction, cumulative
`concentration-effect curves (2 nM-60 mM) to 5-HT were
`constructed in the presence of ascorbate 200 mM, cocaine 6
`mM, indomethacin 2.8 mM, ketanserin 1 mM and prazosin
`1 mM.
`
`10
`
`20
`
`30
`
`40
`
`50
`
`Following a 45—60 min wash period, cumulative
`concentration-effect curves to the test compounds or 5-HT
`(as a time match control) were constructed in the presence
`of ascorbate,
`indomethacin, cocaine, ketanserin and pra-
`zosm.
`
`In this screen the compounds of Example 2, 5, 6, 15, 17,
`24, 25, 26, 28 and 29 had ECso’s in the range 0.04 to 15.
`
`Example 1
`
`60
`
`3-Amino-6-cyano-1,2,3,4-tetrahydrocarba701e hydrochlo-
`ride
`
`A solution of 4-aminocyclohexanol hydrochloride (6.08
`g, 0.04 mole) in water (60 ml) was brought to pH 8 with
`aqueous sodium bicarbonate solution. N-carbethoxy-
`phthalimide (8.76 g, 0.04 mole) was added followed by
`tetrahydrofuran (until homogenous solution was obtained).
`The clear solution was stirred at room temperature over-
`night. During this time a white solid was precipitated. The
`tetrahydrofuran was removed in vacuo and the remaining
`
`MYLAN(Pitav) 037196
`
`
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`Case 1:14-cv-02758-PAC Document 69-4 Filed 06/10/15 Page 7 of 15
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`5,827,871
`
`9
`aqueous solution was extracted with ethyl acetate until the
`solution was clear. The ethyl acetate extracts were
`combined, washed with water, dried (MgSO4) and concen-
`trated to give 4-phthalimido cyclohexanol as a white solid
`(7.1 g).
`A solution of 4-phthalimido cyclohexanol (7.1 g, 0.029
`mole) in dichloromethane (250 ml) was treated with pyri—
`dinium chlorochromate (8.6 g, 0.04 mole) and the resulting
`dark mixture was stirred at room temperature overnight.
`Diethyl ether (50 ml) was added and the mixture filtered
`through keiselguhr. The filtrate was concentrated in vacuo
`and the residue purified by column chromatography (SiOz;
`CHCl3/EtOAc) to give 4—phthalimido cyclohexanone as a
`white solid (6.4 g).
`4-Cyanophenyl hydrazine hydrochloride (4.41 g, 0.026
`mole) was dissolved in acetic acid (100 ml) and sodium
`acetate (2
`was added. 4-Phthalimido cyclohexanone (6.4
`g, 0.026 mole) was added and the mixture heated under
`reflux overnight. The solvent was removed in vacuo and the
`residue triturated with methanol to give 3-phthalimido-6-
`cyano-1,2,3,4-tetrahydrocarbazole as a beige solid, (5.3 g).
`Asuspension of the above product (1 g) in ethanol (40 ml)
`was treated with hydrazine in water (10 ml). The reaction
`mixture was stirred at room temperature overnight during
`which time the reactants dissolved. The solvent was
`removed in vacuo and the residue partitioned between
`aqueous potassium carbonate and ethyl acetate. The ethyl
`acetate solution was washed with water, dried and concen-
`trated in vacuo to give 3—amino—6—cyano—1,2,3,4—
`tetrahydrocarbazole as a beige solid (500 mg). This product
`was converted into the hydrochloride salt to give the title
`compound, mp 289° C. (dec.).
`1H NMR [250 MHz, 03301)] a 1.98—2.18 (1H, m),
`2.25—2.40 (1H, m), 2.77 (1H, dd), 2.98 (2H, m), 3.22 (1H,
`dd), 3.68 (1H, m), 7.34 (1H, d), 7.43 (1H, d), 7.82 (1H, s).
`Example 2
`
`3-An1ino-6-carboxamido-1,2,3,4-tetrahydroearbazole
`hydrochloride
`The product of Example 1 (400 mg) was dissolved in
`tetrahydrofuran, and di—t—butyl dicarbonate (500 mg) was
`added. The mixture was stirred at room temperature over—
`night. The solvent was removed in vacuo and the residue
`purified by column chromatography (SiOZ; CHCl3/EtOAc)
`to give 3-t-butyloxycarbonylamino-6-cyano-1,2,3,4-
`tetrahydrocarbazole (40 mg).
`Amixture of the above product nitrile (440 mg), aqueous
`hydrogen peroxide (30%, 0.5 m1) and sodium hydroxide (aq)
`(20%, 0.5 ml) in methanol (25 ml) was stirred at room
`temperature overnight. Sodium metabisulphite 100 mg)
`was added and the solvent removed in vacuo. The residue
`
`was dissolved in ethyl acetate and the ethyl acetate layer was
`separated, dried and concentrated in vacuo to give a gummy
`solid which was purified by column chromatography (SiOz;
`CHCl3/ EtOAc)
`to give 3-t-butyloxycarbonylamino-6-
`carboxamido-1,2,3,4-tetrahydrocarbazole as a white solid
`(400 mg), mp 270° C. (dec).
`The above product (400 mg, 0.0012 mole) was dissolved
`in dioxan (100 ml) and HCl gas was bubbled through the
`solution for 20 minutes. During this time a white solid was
`precipitated. Excess hydrogen chloride was swept from the
`solution by bubbling through N2, and the solid product,
`3-amino-6-carboxamido-1,2,3,4-tetrahydrocarbazole hydro-
`chloride was collected by filtration, washed with diethyl
`ether and dried to give the title compound as a white solid
`(300 mg). mp. 270 (dec).
`1H NMR [250 MHz, DMSO—d"] 8 1.96 (1H, m),
`2.16—2.30 (1H, m), 2.74 (1H, dd), 2.85 (2H, m), 3.12 (1H,
`
`10
`dd), 1 signal obscured by H20 at ca. 3.6, 7.08 (1H, brd.s),
`7.27 (1H, d), 7.61 (1H, d), 7.87 (1H, brd.s), 7.99 (1H, s),
`8.39 (3H, brd.s).
`
`Example 3
`
`3-Amino-6-methoxy-1,2,3,4-tetrahydrocarbazole hydro-
`chloride
`Reaction of 4-methoxyphenyl hydrazine hydrochloride
`(0.87 g, 5.0 mmol) with 4-phthalimido-cyclohexanone (1.22
`g, 5 .0 mmol) in ethanol (20 ml) heated under reflux for 2 hr,
`followed by cooling and removal of the precipitated solid by
`filtration gave 3-phthalimido-6-methoxy-1,2,3,4-
`tetrahydrocarbazole (1.62 g).
`The above product (1.57 g, 4.5 mmol) was suspended in
`ethanol (100 ml) and treated with hydrazine hydrate (23 ml)
`while stirring at ro