(12) United States Patent
`US 7,112,565 B2
`(10) Patent N0.:
`
`
` Sawai et a]. (45) Date of Patent: *Sep. 26, 2006
`
`USOO7112565B2
`
`(54) STABILIZED PHARMACEUTICAL
`COMPOSITION IN LYOPHILIZED FORM
`
`(75)
`
`Inventors: Seiji Sawai, Takarazuka (JP); Akihiro
`Kasai, lkoma (JP); Kazumi Ohtomo,
`Ibaraki (JP)
`
`(73) Assignee: Astellas Pharma Inc., Tokyo (JP)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 307 days.
`
`This patent is subject to a terminal dis-
`claimer.
`
`(21) App]. N0.: 10/772,281
`
`(22) Filed:
`
`Feb. 6, 2004
`
`(65)
`
`Prior Publication Data
`
`US 2004/0157769 A1
`
`Aug. 12, 2004
`
`Related U.S. Application Data
`
`(62) Division of application No. 09/786,125, filed as appli-
`cation No. PCT/JPOO/04381 on Jun. 29, 2000, now
`Pat. No. 6,774,104.
`
`(30)
`Jul. 1, 1999
`
`Foreign Application Priority Data
`(JP)
`................................. 11/187713
`
`(51)
`
`Int. Cl.
`(2006.01)
`A61K 38/00
`(2006.01)
`A61K38/12
`(52) U.S. Cl.
`................... 514/9; 514/2; 514/11; 514/15;
`530/317; 530/323
`(58) Field of Classification Search ..................... None
`See application file for complete search history.
`
`(56)
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`12/1994 Iwamoto et 31.
`5,376,634 A
`10/1996 Ohki et a1.
`5,569,646 A
`8/1999 Floyd et a1.
`5,942,510 A
`3/2001 Ueda et 211.
`6,207,434 Bl
`6/2002 Kanasaki et a .
`6,399,567 B1
`8/2004 Sawai et al.
`................... 514/9
`6,774,104 B1 *
`FOREIGN PATENT DOCUMENTS
`3-193735
`8/1991
`
`
`
`JP
`
`JP
`JP
`JP
`
`JP
`W0
`wo
`WO
`WO
`
`3-240727
`6-51641
`9-301997
`
`10-507174
`WO 96/11210
`wo 97/39763
`VVO 00/51564
`VVO 00/51567
`
`10/1991
`7/1994
`11/1997
`
`7/1998
`4/1996
`10/1997
`9/2000
`/2000
`
`* cited by examiner
`
`Primary ExamineriB, Dell Chism
`(74) Attorney, Agent, or Firingoblon, Spivak, McClelland,
`Maier & Neustadt, PC.
`
`(57)
`
`ABSTRACT
`
`A stabilized pharmaceutical composition in lyophilized form
`comprising: a cyclic polypeptide compound represented by
`the general formula (I):
`
`R2
`
`011
`
`HO—S—O
`
`110
`
`wherein R1 is a hydrogen atom or an acyl group and R2 and
`R3 are, the same or different, a hydrogen atom or a hydroxyl
`group, or a salt thereof and the stabilizer.
`
`23 Claims, No Drawings
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`1
`STABILIZED PHARMACEUTICAL
`COMPOSITION IN LYOPHILIZED FORM
`
`This application is a divisional of US, Ser. No. 09/786,
`125, filed Mar. 1,2001, now US Pat. No. 6,774,104, which
`is the national—stage under 35 U.S.C. §371 of PCT/JPOO/
`04381, filed Jun. 29, 2000. This application also claims
`priority to JAPAN 11/187713, filed Jul. 1, 1999.
`TECHNICAL FIELD
`
`The present invention relates to a stabilized phannaceu-
`tical composition in lyophilized fomi containing a cyclic
`polypeptide compound. More particularly,
`the present
`invention relates to a stabilized pharmaceutical composition
`in lyophilized form containing a cyclic polypeptide com-
`pound or its phamiaceutically acceptable salt and a stabi-
`lizer.
`The cyclic polypeptide compound ofthe present invention
`is represented by the general formula (I):
`
`(I)
`
`OH
`
`HO—S—O
`
`HO
`
`10
`
`15
`
`30
`
`35
`
`4o
`
`45
`
`wherein R1 is a hydrogen atom or an acyl group and R2 and
`R3 are, the same or different, a hydrogen atom or a hydroxyl
`group. The compound has an antimicrobial activity, particu-
`larly an antifungal activity and a [3—1,3—glucan synthase
`inhibiting action, and is useful for preventing and treating ,
`various kinds of infectious diseases including Pneumacystis
`mrinii infection, e,g., carinii pneumonia.
`
`BACKGROUND ART
`
`Among the cyclic polypeptide compounds represented by
`the above formula (I), a compound wherein R1 is a hydrogen
`atom and R2 and R3 are hydroxyl groups and a compound
`wherein R1, R2 and R3 are hydrogen atoms are obtained by
`a fermentation process disclosed by European Patent No.
`0462531 and processes disclosed by WO97/32975 and by
`WO97/47738. A compound wherein R1 is an acyl group and
`its production process are disclosed by US. Pat. Nos.
`5,376,634 and 5,569,646 and VJO96/11210 and VVO99/
`40108.
`The cyclic polypeptide compounds (I) and their salts are
`generally unstable to light, humidity, acids, heat and the like.
`
`6O
`
`65
`
`2
`Therefore, desired is development of oharmaceutieal prepa—
`rations in which the cyclic polypeptide compounds and their
`salts are stabilized.
`
`DISCLOSURE OF INVENTION
`
`
`
`The present invention provides a stabilized pharmaceuti—
`cal composition in lyophilized form containing a cyclic
`polypeptide compound (I) or its phannaceutically accept-
`able salt and a stabilizer.
`
`The “acyl group” for R1 in the formula (1) representing the
`cyclic polypeptide compound of the present invention is
`now explained. In the context of the present specification,
`“lower” means having one to six carbon atoms unless
`otherwise indicated.
`
`As examples of the acyl group, may be mentioned ali—
`phatic acyl groups, aromatic acyl groups, aromatic—aliphatic
`acyl groups and heterocyclic acyl groups derived from
`aliphatic, aromatic, aromatic-aliphatic and heterocyclic car-
`boxylic acids.
`Examples of the aliphatic acyl groups include lower or
`higher alkanoyl groups such as formyl, acetyl, propanoyl,
`butanoyl, 2—methylpropanoyl, pentanoyl, 2,2—dimethylpro—
`panoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl,
`undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, penta-
`decanoyl,
`hexadecanoyl,
`heptadecanoyl,
`octadecanoyl,
`nonadecanoyl, icosanoyl, etc; cycloalkanoyl groups such as
`cyclopentanoyl and cyclohexanoyl;
`lower alkoxycarbonyl
`groups such as methoxycarbonyl, ethoxycarbonyl, t-butoxy-
`carbonyl,
`t-pentyloxycarbonyl, heptyloxycarbonyl,
`etc.;
`lower alkanesulfonyl groups
`such as methanesulfonyl,
`ethanesulfonyl, etc.; lower alkoxysulfonyl groups such as
`methoxysulfonyl, ethoxysulfonyl, etc.; and the like.
`Examples of the aromatic acyl groups include aroyl
`groups such as benzoyl, toluoyl, naphthoyl and the like.
`Examples of the aromatic-aliphatic acyl groups include
`ar(lower)alkanoyl groups such as phenyl(C14:6)alkanoyl
`(e.g., phenylacetyl, phenylpropanoyl, phenylbutanoyl, phe—
`nylisobutanoyl, phenylpentanoyl, phenylhexanoyl, etc.),
`naphthyl(Cl7C6)alkanoyl
`(e.g., naphthylacetyl, naphthyl-
`propanoyl, naphthylbutanoyl, etc.) and the like; ar(lower)
`alkenoyl group such as phenyl(C34:6)alkenoyl (e.g., phe—
`nylpropenoyl.
`phenylbutenoyl,
`phenylmethacryloyl,
`phenylpentenoyl, phenylhexenoyl, etc.), naphthyl(C37C6)
`alkenoyl (e.g., naphthylpropenoyl, naphthylbutenoyl, etc.
`and the like;
`ar (lower)alkoxycarbonyl groups such as phenyl(C17C6)
`alkoxycarbonyl (e.g., benzyloxycarbonyl, ete.), fluorenyl
`(C17C6)alkoxycarbonyl (e.g., fluorenylmethoxycarbonyl,
`etc.) and the like;
`aryloxycarbonyl groups such as phenoxycarbonyl, naph-
`thoxycarbonyl, etc.;
`aryloxy(lower)alkanoyl groups such as phenoxyacetyl, phe-
`noxypropionyl, etc.;
`arylcarbamoyl groups such as phenylcarbamoyl, etc;
`arylthiocarbamoyl groups such as phenylthiocarbamoyl,
`etc.;
`arylglyoxyloyl groups such as phenylglyoxyloyl, naphthylg—
`lyoxyloyl, etc.;
`arylsulfonyl groups which may be optionally substituted by
`a lower alkyl group such as phenylsulfonyl, p-tolylsulfo-
`nyl, etc.; and the like.
`Examples of the heterocyclic acyl groups include hetero—
`cyclic carbonyl groups such as thenoyl, furoyl, nicotinoyl,
`etc.;
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`10
`
`15
`
`30
`
`35
`
`3
`heterocyclic(lower)alkanoyl groups such as heterocyclic
`acetyl. heterocyclic propanoyl, heterocyclic butanoyl,
`heterocyclic pentanoyl, heterocyclic hexanoyl, etc.;
`heterocyclic(lower)alkcnoyl groups such as heterocyclic
`propenoyl, heterocyclic butenoyl, heterocyclic pentenoyl,
`heterocyclic hexenoyl, etc.;
`heterocyclic glyoxyloyl and the like.
`The acyl group for R1 may have one or more suitable
`substituent(s). Among the above-mentioned examples for
`the acyl groups, an aroyl group which may have one or more
`suitable substitucnt(s) is particularly preferable.
`Examples of suitable substituents in the acyl group
`include a heterocyclic group substituted by an aryl group
`having a lower alkoxy group, a heterocyclic group substi-
`tuted by an aryl group having a lower alkoxy(lower)alkoxy
`group, a heterocyclic group substituted by an aryl group
`having a lower alkoxy(higher)alkoxy group, a heterocyclic
`group substituted by an aryl group having a cyclo(lower)
`alkyloxy group, a heterocyclic group substituted by an aryl
`group having a heterocyclic group, a heterocyclic group
`substituted by a cyclo(lower)alkyl group having a cyclo
`(lower)a11\yl group, a heterocyclic group substituted by an
`aryl group having an aryl group substituted by a lower
`alkoxy(lower)alkoxy and a heterocyclic group substituted ,
`by an aryl group having a heterocyclic group substituted by
`a cyclo(lower)alkyl group.
`Among these examples, preferred are an unsaturated 3— to
`8—membered heteromonocyclic group containing one to two
`oxygen atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl having (C47C6)alkoxy, an Luisaturated
`condensed heterocyclic group containing one to two sulfur
`atom(s) and one to three nitrogen atom(s) and substituted by
`phenyl having (C4436)alkoxy, an unsaturated 3- to 8-mem-
`bered heteromonocyclic group containing one to two sulfur
`atom(s) and one to three nitrogen atom(s) and substituted by
`phenyl having (C17C4)alkoxy(C44:6)alkoxy, an unsaturated
`3— to 8—membered heteromonocyclic group containing one to
`two sulfur atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl having (C1£4)alkoxy(C7iC14)
`alkoxy, a saturated 3- to 8-membered heteromonocyclic
`group containing one to four nitrogen atom(s) and substi—
`tuted by phenyl having (C17C4)alkoxy(C7£14)alkoxy, an
`unsaturated condensed heterocyclic group containing one to
`two sulfur atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl having cyclo(C44:6)alkyloxy, an
`unsaturated condensed heterocyclic group containing one to
`two sulfur atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl, a saturated 3- to 8-membered hetero-
`monocyclic group containing one to two oxygen atom(s) and ,
`one to three nitrogen atom(s), a saturated 3— to 8—membered
`heteromonocyclic group having one to four nitrogen atom(s)
`and substituted by cyclo(C47C6)alkyl having cyclo(C44:5)
`alkyl, an unsaturated 3- to 8-membered heteromonocyclic
`group having one to two sulfur atom(s) and one to three
`nitrogen atom(s) and substituted by phenyl having phenyl
`substituted by (C17C4)alkoxy(C i£4)alkoxy, an unsaturated
`3- to 8-membered heteromonocyclic group containing one to
`two sulfiir atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl having a saturated 3— to 8—membered
`heteromonocyclic group which contains one to four nitrogen
`atom(s) and is substituted by cyclo(C47C5)alkyl, and an
`unsaturated condensed heterocyclic group containing one to
`two sulfur atom(s) and one to three nitrogen atom(s) and
`substituted by phenyl having a saturated 3— to 8—membered
`heteromonocyclic group which contains one to four nitrogen
`atom(s) and has cyclo(C4£6)alkyl.
`
`4o
`
`45
`
`6O
`
`65
`
`4
`Among these, particularly preferred are an isoxazolyl
`group substituted by phenyl having pentyloxy, an imida—
`zothiadiazolyl group substituted by phenyl having penty-
`loxy, a thiadiazolyl group substituted by phenyl having
`methoxyhexyloxy, a thiadiazolyl group substituted by phe-
`nyl having methoxyoctyloxy, a thiadiazolyl group substi-
`tuted by phenyl having methoxyheptyloxy, an imidazothia—
`diazolyl group substituted by phenyl having cyclohexyloxy,
`an imidazothiadiazolyl group substituted by phenyl having
`dimethylmorpholino, a piperazinyl group substituted by
`phenyl having methoxyheptyloxy, a piperazinyl group sub—
`stituted by phenyl having methoxyoctyloxy, a piperazinyl
`group substituted by cyclohexyl having cyclohexyl, a thia-
`diazolyl group substituted by phenyl having phenyl substi-
`tuted by methoxyethoxy, a thiadiazolyl group substituted by
`phenyl having phenyl substituted by methoxybutoxy, a thia—
`diazolyl group substituted by phenyl having phenyl substi-
`tuted by ethoxypropoxy, an imidazothiadiazolyl group sub-
`stituted by phenyl having pipera7inyl
`substituted by
`cyclohexyl, an imidazothiadiazolyl group substituted by
`phenyl having piperazinyl substituted by cyclohexyl, and the
`like.
`
`Accordingly, particularly suitable examples of the acyl
`group of R1 may be a benzoyl group having isoxazolyl
`substituted by phenyl having pentyloxy, a benzoyl group
`having imidazothiadiazolyl substituted by phenyl
`laying
`pentyloxy, a benzoyl group having thiadiazolyl substituted
`by phenyl having methoxyhexyloxy, a benzoyl group iaving
`thiadiazolyl substituted by phenyl having methoxyoc yloxy,
`a benzoyl group having thiadiazolyl substituted by alienyl
`having methoxyheptyloxy, a benzoyl group having imida-
`zothiadiazolyl substituted by phenyl having cyclohexyloxy,
`a benzoyl group having imidazothiadiazolyl substituted by
`phenyl having dimethylmorpholino, a benzoyl group laying
`piperazinyl substituted by phenyl having methoxyhep yloxy,
`a benzoyl group having piperazinyl substituted by ahenyl
`having methoxyoctyloxy, a benzoyl group having pioerazi—
`nyl substituted by cyclohexyl having cyclohexyl, a benzoyl
`group having thiadiazolyl substituted by phenyl
`raving
`phenyl substituted by methoxyethoxy, a benzoyl group hav-
`ing thiadiazolyl substituted by phenyl having phenyl sub-
`stituted by methoxybutoxy, a benzoyl group having tiiadia—
`zolyl substituted by phenyl having phenyl substituted by
`ethoxypropoxy, a benzoyl group having imidazothiadiazolyl
`substituted by phenyl having piperazinyl substitu ed by
`cyclohexyl, a benzoyl group having imidazothiadiazolyl
`substituted by phenyl having piperazinyl substitu ed by
`cyclohexyl, and the like.
`Particularly preferable examples of the acyl groups of R1
`are represented by the formulas:
`
`
`
`O
`
`O
`
`O \\ O “CM—H
`
`N—O
`
`WCH >
`
`/ >_©—o(cnz)70CH3
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`0
`
`o
`
`5
`
`-continued
`
`/ V”
`/
`A \
`N
`s
`
`ogcnmcn,
`
`—NN
`
`/ >—©70(CH2)60CH3>
`
`S
`
`The cyclic polypeptide compounds (I) having the above—
`mentioned acyl groups may be prepared from a compound
`having a hydrogen atom as R1 and hydroxyl groups as R2
`and R3 or a compound having hydrogen atoms as R1, R2 and
`R3 according to the US. Pat. Nos. 5,376,634 and 5,569,646
`and W096/11210 and WO99/40108.
`
`Suitable salts of the cyclic polypeptide compounds (1) are
`soluble in water and pharmaceutically acceptable salts
`including salts With bases and acid addition salts. Such a salt
`
`5
`
`10
`
`15
`
`6
`may be prepared by treating the cyclic polypeptide com—
`pound (1) with an appropriate base or acid according to the
`conventional method.
`
`As salts with bases, may be mentioned salts with inor-
`ganic bases such as alkali metal salts (e.g., sodium salts,
`potassium salts, etc), alkaline earth metal salts (e.g., cal-
`cium salts, magnesium salts, etc.), ammonium salts and the
`like; salts with organic bases such as organic amine salts
`(e.g., triethylamine salts, diisopropylethylamine salts, pyri-
`dine salts, picoline salts, ethanolamine salts, triethanolamine
`salts, dicyelohexylamine salts, N,N'—diben7ylethylenedi-
`amine salts, etc.); and the like.
`As acid addition salts, may be mentioned inorganic acid
`addition salts (e.g., hydrochloridcs, hydrobromidcs, sulfates,
`phosphates, etc.); and organic carboxylic or sulfonie acid
`addition salts (e.g.,
`fonnates, acetates,
`trilluoroacetates,
`maleates, tartrates, fumarates, methnesulfonates, benzene-
`sulfonates,
`toluenesulfonates, ete.). Further, may also be
`mentioned salts with basic or acidic amino acids (e.g., salts
`With arginine, aspartie acid, glutamic acid, etc.).
`The cyclic polypeptide compounds (1) of the present
`invention also include possible conformers and a pair or
`more of stereoisomers such as geometric isomers and optical
`isomers which may exist due to asymmetric carbon atoms.
`The preferable ones of the cyclic polypeptide compounds
`(1) are represented by the following formulas (11) to (V1):
`
`(to be continued on the next page)
`
`(n)
`
`O \\ O Oflmmms
`
`N—O
`
`2\EOKN} EN
`32* 45:
`NaO—S—O
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`-continued
`
`Om
`
`HO 0
`
`NH
`
`mC)
`
`O
`
`CE
`
`Z
`
`Zm
`
`0
`
`)m026¢ ONz E‘o
`Om 0
`
`o
`
`HO
`
`||
`NaO—S—O
`||0
`
`(HI)
`
`(1V)
`
`(V)
`
`\2 VI3/34 0
`
`N—\I
`/
`\
`
`S
`
`0(CH2)70CH3
`
`/ N’N\
`/
`A
`N
`5
`
`O(_’CH2)4CH3
`
`OH
`
`NH
`
`0
`
`HO 0
`
`NH
`
`N
`
`110
`
`0
`
`HN
`
`1-1(,
`
`0
`
`NH
`
`0
`
`N
`
`NH
`
`OH
`
`O
`
`H2N
`
`o
`
`H
`NaO—S—O
`||0
`
`H0
`
`,
`OH
`
`HO O
`
`NH
`
`ch
`
`o
`
`N
`
`NH
`o
`
`HO
`
`O
`
`IN
`
`o
`
`0H
`
`CH3
`
`011
`
`0
`
`011
`
`CH3
`
`0
`
`N
`
`o
`||
`NaO—S—O
`
`IIo
`
`NOH
`
`0
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`-continued
`
`10
`
`(VI)
`
`—\I
`
`7
`
`>—QO(:CHZ)6OCH3_
`
`s
`
`.11}
`
`1I-TN
`§«H3
`
`30
`
`35
`
`The most preferable one is represented by the formula
`(II).
`The amount of the cyclic polypeptide compound (I) or its
`pharmaceutically acceptable salt contained in the composi-
`tion for a single unit dosage of the present invention is 0.1
`to 400 mg, more preferably 1
`to 200 mg, still more prefer-
`ably 10 to 100 mg, specifically 10, 15, 20, 25, 30, 35, 40, 45,
`50, 55, 60, 70. 75, 80, 85, 90, 95 and 100 mg.
`As the stabilizer, may be mentioned polysaccharides,
`disaccharides, sodium chloride and a combination thereof.
`Examples of the polysaccharide are dextran, starch, cel—
`lulose and hyaluronic acid; and examples of the disacharide
`are lactose, maltose and sucrose. The polysaccharide or
`disaccharidc contained in the pharmaceutical composition of
`thc prcscnt invcntion may bc ot-monohydratc, ot-anhydridc,
`B-anhydride or a combination thereof.
`The amount of the stabilizer used in the pharmaceutical
`composition of the present
`invention should be at
`least
`sufficient for stabilizing the cyclic polypeptide compound (I)
`or its pharmaceutically acceptable salt in the composition. In
`order to stabilize the cyclic polypeptide compound (I), one
`part by weight of the stabilizer with respect to one part by
`weight of the cyclic polypeptide compound (I) or its phar-
`maceutically acceptable salt in the present composition is
`suflicient at least. The stabilizer may also serve as a carrier .
`or an excipient. Thus the use amount of stabilizer does not
`have a particular upper limit and may be determined in
`consideration of the weight or volume of the composition
`with respect to a unit dose of the compound and the like.
`However, such amount
`is prcfcrably 0.4 to 50 parts by
`weight, more preferably 0.6 to 20 parts by weight, still more
`preferably 0.8 to [0 parts by weight with respect to one part
`by weight of the cyclic polypeptide compound (I) or its
`pharmaceutically acceptable salt, though it varies depending
`upon the kind and the used amount of the cyclic polypeptide
`compound (I) or its pham1aceutically acceptable salt,
`its
`preparation form and/or the like. Specifically,
`it
`is more
`preferable that 1 to 20 parts, still more preferably 2 to 10
`parts by weight of thc disaccharidc arc uscd with rcspcct to
`one part by weight of the cyclic polypeptide compound (I)
`or its pharmaceutically acceptable salt. Specifically,
`it is
`more preferable that 0.6 to 20 parts, still more preferably 0.8
`to 10 parts by weight of sodium chloride are used with
`
`respect to one part by weight of the cyclic polypeptide
`compound (I) or its pharmaceutically acceptable salt.
`The pharmaceutical composition of the present invention
`may be produced according to methods known in the art
`with using additives if necessary. Here, Basic Lecture on
`Development ofPharmaceuticals XI 20 Production ofPhar—
`maceuzicals (the second volume) (edited by Kyosuke Tsuda
`and Hisashi Nogami and published by Chizyo Shoten) is
`mentioned for reference. The lyophilized composition may
`be obtained by preparing an aqueous solution of the cyclic
`polypeptide compound (I) or its pharmaceutically accept—
`able salt and the stabilizer, optionally adding a pH adjustor
`(citric acid anhydrous, sodium hydroxidc, etc.) as required
`to attain pH 4.(L7.5, preferably pH 4577.0, and then
`lyophilizing the resulting solution in vial according to a
`conventional method. Thus, the stabilized pharmaceutical
`composition in lyophilized form, when dissolved in purified
`water, preferably gives a solution of pH 4.0 to 7.5, more
`preferably pH 4.5 to 7.0.
`It
`is preferable that
`the thus
`prepared composition in lyophilized form is sealed and
`stored with shading. The lyophilized composition can be
`loadcd in cach Vial in the solution form bcforc lyophilizing
`or in lyophilized powder form after lyophilizing.
`Since the cyclic polypeptide compound is not satisfacto—
`rily stable to humidity, it is necessary that the lyophilized
`composition of the present
`invcntion contains 3.4% by
`weight or less of water, preferably 3.0%, more preferably
`2.0%.
`
`Usually the stabilized pharmaceutical composition in lyo-
`philized form is dissolved in isotonic sodium chloride solu—
`tion as required and used as an injection solution. The
`pharmaccutical composition of the present invcntion may be
`used as an injection preparation which requires some com-
`pounding before use,
`
`BEST MODE FOR CARRYING OUT THE
`INVENTION
`
`The present invention is now described in further detail by
`way of examples and test examples, which should not be
`construed to limit
`the scope of the invention.
`In the
`examples, the compounds of formula (II) to WI) are referred
`to as Compounds (II) to (VI), respectively.
`
`AMNEAL EX. 1006
`
`4o
`
`45
`
`6O
`
`65
`
`AMNEAL EX. 1006
`
`

`

`11
`EXAMPLE 1
`
`12
`EXAMPLE 9
`
`US 7,112,565 B2
`
`Lyophilized compositions each containing 50 mg ofCom-
`pound (V) are obtained in the same manner as in Example
`1 except that 50 g of Compound (V) is used instead of
`Compotmd (11) and 50 g of soditun chloride is used instead
`of lactose.
`
`EXAMPLE 10
`
`Lyophilized compositions each containing 10 mg ofCom—
`pound (V1) are obtained in the same mamier as in Example
`1 except that 10 g of Compound (VI) is used instead of
`Compotmd (II) and 100 g of dextran is used instead of
`lactose.
`
`Test Example 1
`
`ElIect of Stabilizer in Stabilizing Lyophilized Compositions
`of Compotmd (11)
`10 mg of Compound (11) and, as a stabilizer. 100 mg of
`lactose or 9 mg of sodium chloride were dissolved com—
`pletely in 1 ml of water. The resulting solutions were
`lyophilized and maintained at 70° C. in glass vials. Nine
`days after, the resulting compositions were tested on their
`appearance,
`the residual amount of Compound (11), and
`others. As a control, used was a solution of Compound (II)
`without any stabilizers. The results are shown in Table 1.
`
`TABLE 1
`
`Stabilizers Test Items
`
`0 hours
`
`After 9 days
`
`Control:
`nil
`
`Lactose
`(100 mg)
`
`4o
`
`Sodium
`chloride
`(9 mg)
`
`Appearance
`pH*
`Residual amount (%)
`Water content (%)
`Appearance
`pH*
`Residual amount 1%)
`Water content (%)
`Appearance
`pH*
`Residual amount 1%)
`Water content (%)
`
`Slightly yellow mass
`White mass
`2.7
`7.1
`8.0
`100.0
`i
`1.3
`White mass White mass
`6.4
`6.1
`100.0
`99.5
`1.0
`White mass White mass
`6.7
`6.3
`100.0
`75.9
`0.7
`7
`
`Compound (H) 25 g
`Lactose 200 g
`anhydrous Citric acid in a suitable amount
`Sodium hydroxide in a suitable amount
`Lactose was dissolved in purified water (2000 ml) under
`heating below 500 C. After cooling below 20° C, the lactose
`solution was added with Compound (11) avoiding bubbling
`under gently stirring. After adding 2% aqueous citric acid
`solution (9.5 ml), the solution was added with 0.4% aqueous
`sodium hydroxide solution (about 24 ml) to adjust pH 5.5
`followed by diluting with purified water to make a given
`volume (2500 ml). The resulting solution was dispensed into
`1,000 vials of 10 mL volume, 2.5 ml per Vial. The solution
`in the respective vials was lyophilized by using the lyo-
`philizer (RI.-603BS manufactured by Kyowa Shinku Co.,
`Ltd) by the conventional method to obtain lyophilized
`compositions each containing 25 mg of Compound (11).
`EXAMPLE 2
`
`Lyophilized compositions each containing 50 mg of Com—
`potmd (11) were obtained in the same maimer as in Example
`1 except that the amount of Compound (II) used was 50 g.
`EXAMPLE 3
`
`Lyophilized compositions each containing 25 mg of Corn-
`
`pound (II) are obtained in the same manner as in Example
`1 except that 150 g of maltose is used instead of lactose.
`EXAMPLE 4
`
`Lyophilized compositions each containing 50 mg of Com—
`
`pound (II) are obtained in the same manner as in Example
`1 except that the amount of Compound (II) used is 50 g
`instead of 25 g and 250 g of sucrose is used instead of
`lactose.
`
`10
`
`15
`
`30
`
`35
`
`EXAMPLE 5
`
`Lyophilized compositions each containing 25 mg of Com-
`
`pound (II) are obtained in the same manner as in Example
`1 except that 25 g of sodium chloride is used instead of
`lactose.
`
`EXAMPLE 6
`
`*pH of reconstituted solutions of compositions in 1 m1 ofwatcr
`
`45
`
`Test Example 2
`
`Lyophilized compositions each containing 10 mg of Com—
`
`potmd (II) are obtained in the same manner as in Example
`1 except that the amount of Compound (II) used is 10 g
`instead of 25 g and 100 g of dextran is used instead of 7
`lactose.
`
`EXAMPLE 7
`
`The similar tests were conducted in the same manner as
`
`in Test Example 1 except that 100 mg of maltose, 50 mg of
`sucrose or 50 mg of glucose was used as a stabilizer. The
`results are shown in Table 2.
`
`(to be continued on the next page)
`
`TABLE 2
`
`Lyophilized compositions each containing 25 mg of Com—
`
`pound (III) are obtained in the same manner as in Example
`1 except that 25 g of Compound (III) is used instead of
`Compound (II) and 200 g of maltose is used instead of
`lactose.
`
`EXAMPLE 8
`
`Lyophilized compositions each containing 10 mg of Com-
`
`pound (IV) are obtained in the same manner as in Example
`1 except that 10 g of Compound (IV) is used instead of
`Compound (II) and the amount of lactose used is 100 g
`instead of 200 g.
`
`Stabilizers
`
`Test Items
`
`0 hours
`
`After 9 days
`
`Control: nil
`
`60
`
`65
`
`Maltose
`(100 mg)
`
`Sucrose
`(50 mg)
`
`Appearance
`pH*
`Residual amount (%)
`Water content (%)
`Appearance
`pH*
`Residual amount (%)
`Water content (%)
`Appearance
`pH"
`Residual amount (%)
`Water content (%)
`
`White mass
`6.8
`100.0
`3.3
`White mass
`7.3
`100.0
`0.9
`White mass
`6.9
`100.0
`1.1
`
`White mass
`5.4
`<75.0
`i
`White mass
`6.7
`98.6
`i
`White melt
`7.0
`8.2.4
`i
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`US 7,112,565 B2
`
`TAELE 2-continued
`
`Stabili7ers
`Glucose
`(50 mg)
`
`Test Items
`Appearance
`pl-1*
`Residual amount (‘34:)
`Water content (%)
`
`0 hours
`White melt
`6.9
`100
`4.3
`
`After 9 days
`Brown melt
`3.6
`1.1
`
`7
`
`*pH ofrcconstitutcd solutions of composttions in 1 ml ofwatcr
`
`the lyophilized
`1 and 2,
`As is obvious from Tables
`composition of Compound (11) and lactose, sodium chloride,
`maltose or sucrose was significantly stable as compared with
`the one not containing any stabilizers or containing other
`stabilizers.
`
`Test Example 3
`
`Dependence of the Stability of Lyophilized Compositions of
`Compound (11) upon the Amount of Lactose Added
`Tests were carried out in the same manner as in Test
`example 1 except that 20 mg, 50 mg, 100 mg or 200 mg of
`lactose were added as a stabilizer. Table 3 shows the results
`of tests by observation of the appearance of compositions,
`the residual amount of Compound (11), the appearance of
`reconstituted solutions of compositions in 1 ml of water, and
`the like. Incidentally, it took 15 seconds to reconstitute the
`compositions in 1 ml of water.
`
`TABLE 3
`
`Amount
`of
`lactose
`added
`Test Items
`(mg)
`20 Aspearance
`
`
`
`Color"
`C arity"
`pH*
`Residual amount (9/0)
`Total impurities (9/0)
`Water content (9%)
`50 Aapearanee
`Color"
`C arity*
`pH”
`Residual amount (:%)
`Total impurities (9/0)
`Water content (%)
`100 Aopearance
`Color*
`C arity"
`pH*
`Residual amount (9/0)
`Total impurities (9/0)
`Water content (%)
`200 Aopearance
`Color‘
`C arity>F
`pH*
`Residual amount (We)
`'lotal impurities (9/0)
`Water content (9%)
`
`Alter 3
`months at
`40° C. and a
`After
`75%
`9 days
`humidity
`at 70° C.
`0 hours
`Slightly White mass
`White
`yellow
`mass
`1118.55
`Colorless
`Colorless White
`Clear
`Not clear Clear
`6.09
`3.03
`6.57
`100.0
`88.09
`100.0
`3.44
`12.3
`3.99
`1.2
`i
`White
`White
`mass
`mass
`Colorless Colorless
`Clear
`Clear
`6.57
`5.56
`100.0
`96.7
`3.32
`7.37
`0.5
`i
`White
`White
`mass
`mass
`Colorless Colorless
`Clear
`Clear
`6.58
`6.08
`100.0
`96.7
`3.43
`7.08
`0.3
`i
`White
`White
`mass
`mass
`Colorless Colorless
`Clear
`Clear
`6.78
`5.70
`100.0
`96.1
`3.40
`7.30
`0.3
`i
`
`i
`White mass
`Colorless
`Clear
`6.26
`99.8
`4.21
`i
`White mass
`Colorless
`Clear
`5.80
`99.6
`3.96
`
`i
`White mass
`Colorless
`Clear
`5.36
`99.6
`4.35
`7
`
`*Color, clarity and pH olreconstitutcd solutions of compositions in 1 ml
`ofwatcr
`
`10
`
`15
`
`20
`
`30
`
`35
`
`4o
`
`45
`
`60
`
`65
`
`14
`As is obvious from Table 3, the lyophilized compositions
`of 10 mg of Compound (11) and various amount of lactose
`had no problem in their stability.
`
`Test Example 4
`
`Stability of Lyophilized Compositions of 200 mg of Lactose
`and Compound (11) in Vial
`Tests were carried out in the same manner as in Test
`Example 1 except that 12.5 mg, 25 mg, 50 mg, 75 mg or 100
`mg of Compound (11) were used with 200 mg of lactose.
`Table 4 shows the results of the tests on the residual amount
`of Compound (11) in the resulting compositions and the like.
`Regarding all the compositions, their appearance is a white
`mass, the time for reeonstitutional dissolution was 15 sec-
`onds, and the color and the clarity of reconstituted solutions
`of the compositions were colorless and transparent.
`
`TABLE 4
`
`Amount
`Afier 3
`of
`months
`Com-
`at 40° C.
`pound
`and
`(11)
`a 75%
`added
`humid-
`Test
`(mg)
`ity
`12.5 3’7
`6.08
`{esi ua amount ('%)
`99.6
`Tota imourities (’96)
`2.71
`Water content 11%)
`3**
`6.14
`{esi ua amount (_%)
`101.2
`'ota imourities (_%)
`2.68
`Water content 1%)
`i
`3 . 7
`6.00
`{csi ua amount (”/0
`100.5
`Tota imauritics (%)
`2.74
`Water content 11%)
`i
`3
`*
`6.04
`{esi ua amount (%
`99.0
`Tota imaurities (%)
`2.76
`Water content (“/b)
`i
`37*
`5.85
`{esi ua amount (%)
`99.5
`Tota imwurities (%)
`2.79
`Water content (“/ii)
`i
`
`
`
`
`
`
`
`
`
`25
`
`50
`
`75
`
`100
`
`tems
`
`
`
`
`
`
`
`
`
`After 9 Afier 21
`days at
`days at
`70° C.
`60° C.
`6.15
`6.31
`98.1
`97.5
`3.95
`3.75
`6.07
`6.11
`99.3
`98.2
`4.03
`3.49
`i
`i
`5.99
`6.00
`97.9
`97.3
`3.95
`3.68
`i
`i
`5.95
`5.96
`98.1
`97.7
`4.14
`3.83
`i
`i
`5.92
`5.88
`97.8
`96.7
`4.15
`3.92
`i
`i
`
`0 hours
`6.63
`00.0
`2.24
`1.3
`6.37
`00.0
`2.25
`1.1
`6.26
`00.0
`2.25
`1.2
`6.13
`00.0
`2.28
`0.9
`6.03
`00.0
`2.46
`1.3
`
`
`
`>"pH of reconstituted solutions of compositions in 5 ml ofpnrified water
`
`As is obvious from Table 4, all the lyophilizcd composi-
`tions were stable.
`
`Test Example 5
`
`Stability Test The pharmaceutical compositions obtained in
`Examples 1 and 2 were stored at room temperature. After 18
`months, the residual ratio of Compound (11) was 98% in all
`the compositions.
`
`Test Example 6
`
`Dependence of the Stability of Lyophilized Compositions of
`Compound (11) upon the pH Value of the Solution of the
`Composition before Lyophilizing
`10 mg of Compound (11) and, as a stabilizer, 100 mg of
`lactose were dissolved completely in 1 ml of citrate-NaOH
`buffer having dilTerent pH value between pH 4.0 to 7.0. The
`resulting solutions having dilTerent pH values were lyo-
`philized and maintained at 700 C. in glass vials. Nine days
`
`AMNEAL EX. 1006
`
`AMNEAL EX. 1006
`
`

`

`15
`after, the resulting compositions were tested on their pH and
`the residual amount of Compound (11). The results are
`shown in Table 5.
`
`US 7,112,565 B2
`
`
`
`TABLE 5
`
`pH of the solution
`of the composition
`before lyophilizing
`5.0
`5.5
`6.0
`
`4.0
`
`4.5
`
`3.9
`0.2
`
`4.4
`0.2
`
`4.8
`0.3
`
`5.4
`0.3
`
`5.8
`0.3
`
`100
`
`5
`
`6.5
`
`6.4
`0.4
`
`7.0
`
`10
`
`6.8
`0.3
`
`4.0
`94.4
`
`4.5
`95.9
`
`4.9
`97.4
`
`5.4
`98.5
`
`5.8
`97.7
`
`6.4
`96.9
`
`6.8
`95.8
`
`0
`hours
`
`9
`days
`
`pH*
`Water
`content (%)
`Residual
`amount (%)
`pH’F
`Residual
`amount (0/0)
`
`16
`
`iMicrospomm
`
`Acremonium;
`Absidia (e.g., Absidia convmbifera, etc);
`Aspergillus (e.g., Aspergillus clavalus, Aspergillusflavus,
`Aspergillus fumigatus, Aspergillus nidulans, Aspergillus
`niger, Aspergillus terreus, Aspergillus versicalar, etc);
`Blastomyces (e.g., Blastumyces dermatitidis, etc);
`Candida (e.g., Candida albicans, Candida glabrata, Can—
`dida guilliermondii, Candida kefvr, Candida krusei, Can—
`dida parapsilosis, Candida stellaloides. Candida lropicalis,
`Candida utilis, etc.);
`Cladosporium (e.g., Cladosporium Wichoides, etc);
`Coccidioides (e.g., Coccidioides immiiis, etc);
`Cryptococcus (e.g., Cnpiococcus neoformans, etc);
`Cunninghamella (e.g., Cunninghamella elegans, etc);
`Dermalophyle;
`Exophiala (e.g., Exophiala dermaliiidis, Exop/ziala spin—
`ifi’m, etc);
`Epidermophyton (e.g., Epidermophyton floccosam, etc);
`Fonsecaea (e.g., Fonsecaea pedrosoi, etc);
`Fasariam (e.g., Fusarium solani, etc);
`Gentrichum (e.g., Gentrichum candiddum, etc);
`Histoplasma (e.g., Histoplasma capsulatum var. capsula—
`nim, etc);
`Malassezia (e.g., Malassezia fuifur, ec);
`Microsporum (e.g., Alicrosporum canis,
`gypseum, etc);
`Mucor;
`Paracoccidioides (e.g., Paracoccidioides brasiliensis,
`etc);
`Penicillimn (e.g., Penicillin”: mamefié’i, etc);
`Phia/op/Iora;
`Pneumocyszis (e.g., Pneumocystis carinii, etc);
`Pseudallescheria (e.g., Pseuda/lescheria boydii, etc);
`Rhizopus (e.g., Rhizopus microsporus var. rhizopodifor—
`mis, R/zizopus oryzae, etc);
`Sacchammyces (e.g., Saccharnmyces cerevisiae, etc);
`Scopulariopris;
`Spuruthrix (e.g., Sporathrix schenchii, etc);
`Triclzophylon