United States Patent 11'1
`DcslI i ct al.
`
`[54[ PROTEIN STABILIZE!)
`PHARMACOLOGICALLY ACTIVE AGENTS,
`METHODS Fon TUE "REI'ARATION
`TH .. :REOF ANn METHODS FOR THE USE
`THEREOF
`
`[75J
`
`invcntors: Nell 1'. l)(>sal, Los AIl gcles; C hunlin
`11m, Beverly Hills: Andrew Y:ml,:.
`Rosemead; u-slk> Louie, Montebello;
`T iltnl] ZI1l'ng; Zhiwcn VilO, both of
`Culver Cily: Putric!, Soon-Shiong. Los
`Angeles, all of Calif.; Shtomo
`Mab'(hl~ i , Jtrusalcm, b rad
`
`[73] A'iSigncc: VI\'IJrll Pllllrma~uticals, Inc .. Santa
`M{)nica, Calif.
`
`[211
`
`Appl. No.: 08/720.756
`
`[22]
`
`Filed :
`
`Oct. I, 1996
`
`Related U.S. Application !lata
`
`Continuation·in-part of aWlical ion No. 08/412.726, Mar.
`29, 1995. Pal. No. 5,560,933, " .. hich is a division of appli·
`catiQll No. 081023.698, Feb. 22, 1993, Pat. No. 5,439/586.
`Int. CI.6
`U.s. CI.
`
`A61K 9/14
`424/489; 424/450; 424/465;
`424/451; 424/439
`............... ......... 4241489, 422.
`Fk'ld of Search
`424/423,475,9.1.9.3,9.32.450,400
`
`IWI
`
`1511
`[52]
`
`1581
`
`[56[
`
`U.S PATENT DOCUMENTS
`
`3,959,457
`4,073,943
`4,247.<106
`4,572,21)3
`
`51l97!> Speake, el .1.
`211978 W,tllind ~I al
`1f19$1 Widdtr 01 a1.
`2119116 F~i n't~in .
`
`(Lisl cOn1inuetl on nexl page.)
`
`FOREIGN PATENT I)OCUMENTS
`
`111111111111111111111111111111 11111 1111111111111111111111111111111111111
`USOO5916596A
`IJatent Number:
`[II]
`[45[ Date or IJatent:
`
`5,916,596
`Jun. 29, 1999
`
`( "'opean Pal. Of!".
`0295 9·H A2 12/1988
`0.391 SIS Al 2/1990 E'JI"p"an Pal. ON.
`
`(Lisl oonlinlloo on nex t page.)
`
`OTI [ER PUnI ,Ir:ATI ONS
`
`Burgess et aI., "Po temial usc 'If albumin micrOlipheres as a
`dru g delivery SySICnl. I. I'reparalion and in vilro rclcas.: o f
`Sleroids,"
`InrernmioMI Journal of Pilarlllilcelllics,
`39:129-136 (1987)
`(I .i~t contimled on neXI page.)
`
`Primary EXllllliner-Ncil S. Levy
`A lli~/(Jm £wllliner-Willialll E. Ik nstoll, Jr.
`Allom ey, t\ gem, or Firm-Dray, Cary, Wa rt: & F rtidenrich;
`Stephen E. Re ite r
`[S7[
`
`AUSTRACf
`
`In aeconl~1JC(: with the prc.""m invention, Ihcrc ~rc pffivided
`compositions and melhods useful for the in vivo delivery o f
`substantially water insol uble pharmacologically active
`agents (such as Ihe anticancer tlrug paditaxel) in whieh the
`phar",ac()l()gi~all y aclive agelll is lIelivereli ill Ihe f()rOI of
`suspended particles coated with prOiein (which acts as a
`stabili:r.ing agent). [n particular, l)rOlein and phaffilacologi.
`cally active agenl in a biocompatibJe diS[lCr.;ing medium are
`subjected to high shear. in the absence o f any convenlional
`surfaclallls, and also in lhe alJscncc of a n ~' polymeric rore
`material for the panicles. The procedure yicld<; panicles wit h
`a tliametn ()f less than abou t 1 minon. The use of spt'dfic
`COmposition and preparalion O)ndilions (e.g., addition of a
`polar solvent to the organic phase), and care ful selection o f
`the proper organic phas.: and phase fradion, enables Ihe
`reprod ucible production of unusually small nanopanieles o f
`less than 200 nm diamctt r, which call be stnile· filtnctl . The
`parlicu lale .<;ySlcm pmdllce(! accord ing 10 Ihe invent; oo can
`be convened into a rt:dispersible dry powder comprising
`nano particles of water·insoluble drug coated with a prole in,
`and free prOlein to which molecules of the pharmacological
`agelll art: lx.ouod. 'Il,is resul lS ill a unique liclive ry system, ill
`which part nf Ihe pharmacologically aClive agent is readilr
`bioavailable (in the fonn of molecules bound 10 Ihe protein),
`and part of the agenl is present within parlicles without any
`polymeric matrix there in.
`
`0 12')61<) A t
`
`I l lqR.~
`
`" uropean Pa l. Off.
`
`31 Claims, 2 Ilnlwing Shtocts
`
`~ r-------------------------~-'
`
`,~
`
`"'"
`~ • ""
`~ • "'"
`E ,
`~ , ~
`1 ,~
`,
`"
`
`- -. . _ - . - - - .... _ . _ _ • _ __ __ -
`T_lrunt PerI(KI
`
`-
`
`- ... ____ - _ _ -
`
`-
`
`- ___ <0
`
`.oo ~~~~' ~~~~----~~~~~~~.
`12t3 1 4 1~t6 17 18 1 9W21llDM~~vu~~3In»~
`
`Days postimplant
`
`Apotex v. Abraxis - IPR2018-00151 , Ex. 1026, p.OI of 19
`
`

`

`5,916,596
`P.d~C 2
`
`u.s. PATENT DOCUMENTS
`6/1987 Goldl>crg CI 01.
`4,671,'JS~
`11191*: Fe;n,le;n ,
`4,718,433
`4,789,550 12/1988 nVl1ll1ld d al
`7/1989 Widdtr 01 a1.
`4,844,882
`5,OS9,6?? 10/1991 Kings/on 01 a1.
`5119'12 Gl'yc. CI 01.
`5,1 10,606
`9/]9'r2 t ;vo"idge oJ "I.
`424/4Rq
`5, 145,684
`5,362,478 11/]994 0. .. ; ri ai, """""" .. ,,' """""" '" 42419
`5,439,686 ~1995 Desa; el ai,
`424/451
`3/1996 Grinslaff el .1.
`424f450
`5,4'JH, 421
`4/1996 Grislaff Ct a1.
`424f./,3
`S,505,9.l2
`.1.
`424/9,322
`4/19% G,in~t.ff d
`5,508,021
`4/1996 Grin,lalr Cl .. I.
`424/9,322
`5,512,268
`5,560,933 10/1996 Soon·Shing cl al.
`424/489
`7/19')7 Grinsloff CI al.
`424f400
`5,650, 156
`5,665,382 W]997 (;nn.laff.I.1.
`424/450
`
`54915 11
`
`FOREIGN PATENT DOCUMENTS
`
`4/ ]990 European 1'lI1. 0 •.
`0361 6n 1\1
`3/ ]9')] European 1'lI1. 0 •.
`04111153AI
`5/199] Europea n 1':11. 01'1.
`0190050IU
`911991 E",opean I'lII Off
`02 13 30J 81
`Fra nc~ ,
`2660556 1011991
`111985 WIPO,
`WO 85.00011
`2/1988 WIPO,
`WO 87/OIm5
`31191*: WII'O ,
`WO 88/01506
`\\'088/07365 10/1988 W[PO ,
`511989 WI1'O ,
`WO 89/03674
`WO 90113285 11/1990 WII'O ,
`wO 'IOf137l'.O 1111990 WII'O,
`WO 9] /15947 10/ 1'191 WIPO
`wo 94/10980
`5/1994 WIPO
`
`0'111£1{ PUllLlCA:1l0NS
`
`Chen el aI., "Com parison of albumin and casein micro(cid:173)
`spheres a,~ a ca rrier for doxonloicin," J. Plwrm. Plwrmacnl.,
`39:978- 985 ( 1987),
`Feinslein (t aI., "Two-Dimensional Con;mast Echocardio(cid:173)
`W'aphy, I. In Vitro Dtvtlopm ~nt amJ Quanlitaliv~ Analysis
`of Echo ConlraSl Agenls," JACG~ 3( I): 14-20 (1984)
`Grinstalf & Susliek, "Nonaqueou~ Liquid Filled Microeap(cid:173)
`sulcs," Polym. "repr" 32:255- 256 (1991).
`Gupla CI aI., "A1bumin min~ph~r~s, Ill. Symhcsis an d
`characterization of microspheres oomaining adriamycin and
`Imernarional ]()lIrrml nf l'lwn/wcellfic.l;
`magnclite,"
`43:167- 177 (1988),
`Ishizaka el al.. "Prepara tion o f Egg Albumin Microcapsules
`and Mi<':rosphcrcs," JVllrlwl uf Phur",,,cl!ulicu/ SciI!IICt"'i,
`70(4):358--363 ( 19tH),
`KlioaooY cI aI., "Amphipathic polyclhylencglycols effec(cid:173)
`livcly prolong the circulalion limc of lipooomcs," FEBS,
`26S( 1):235-237 (1990),
`Koenig & Mcltze r, "£I1'ed of Visrosity on Ihe Si1,C of
`Microhll hhlc~ Generated for IJS(; as Echocardiographic Con(cid:173)
`IraS! Agcnts," JOllrnal of CarriiOl'asclIldr IIflrasonogropil>;
`5(1 ):3-4 (1 986),
`Molecular 13iosy"lcms,
`Invesligator'$ Package"
`
`" AL13 UNEX-Pr~clillical
`
`[nc"
`
`Mosek)' (:t aI., "Microbubb les: A Novel MR Susceplibilily
`Conlm.~t Agcnl." 10th Annual Mooting of Socicly of Mag(cid:173)
`netic Resonance in Medicine (1991).
`SuslLck & Grins/aIT, "Prol..:in MicrOl' ncapsuialiun of Non(cid:173)
`aqueous Liquids," J. Am. Chem. Soc., 112(21):7807-7809
`(1990 )
`Willmon & llarrison. "CharJclerisalion of frecle--d ried
`albumin mierosphcn:s <.'()nlaining Ihe anli-canl't:r drug
`International ]omnal of Pll(lrlllacelitics,
`adrialllycin."
`43:16 1-166 (1988)
`- -, "Serum Albumin Beads: An Injectable, 13iodegrad·
`able System for the Suslained Release of Drugs," Science,
`213(10 ):233--235 (1981),
`UaziJe el. aI., "llody distribu lion o f fully biodegeradable
`J4C}-poly(lat ic acid) naooparticlcs coated w ith albumin
`[
`rats" Biomareriais,
`to
`after parenteral admiostration
`13: 1093-1102 (1992),
`Doury et aI., "lJilalatio nal Properties o f Absorbed Pol y( lJ,
`L- Iactide) and Bovine Serum Aloumin Mono layers althc
`Dichloromethane/Water Interface" Langmuir, 11: 1636-1 644
`(1995).
`Calvo et aI., "Comparative in Vitro l:valuation of Several
`Colloidal SyslemS, Nanllpa r1 iolc", N. nocap~lJlc.~,
`aTliI
`Nanoemulsions, as Ocular Drug Carriers" J. Plwrm. Sci"
`85(5):530-536 (1996)
`Cavalier et a]" "The formation and charao;terization of
`hyrirOC<lrtisonc-llladed poly«( . _ }-Iaoti,k:) microspherc~" J
`Pllarlll. Pllarf/wcoi" 38:249- 253 (1985)
`Kumar el aL "Binding, of Taxolto Iluman Plasma,A1bumin
`arKI-Acid G l ycoprot~in" R<:St'arcl, COIIIII"",icalions in
`Ch<:I1l,clll I'atho/ogy ",,,/ I'f",rmac%g}, 110(3):337-344
`(1993),
`Lee et aL "Serum Albumin Beads: An Injectable, 13inde·
`gradabk Syskm for Ihe Suslaill~d Rclc<tSC of Drugs" Sci(cid:173)
`ellC'~ 2U:233-235 ( 1981).
`l .cucuta 01 aI., "Alhumin m ierospherc.~ as a drug delivery
`sYSlem for e pirubicin: pharmaceulical, pharmacokinetic and
`biological aspt~1s" ImerfUlliViUli JOIlmlil vf PII(tr/llilCelllics,
`41 :213-217 (191:18)
`Livcn;,ideg-Merisko el aI., "FormulBlion and Antilumor
`Activity Evaluation of NanocrYSlaJline SuspensiollS o f
`Poorly Soluble Anlicacer ]) rugs~ Pharmac<:lIIicai Research,
`13(2):272- 278 (19%).
`Mathew el ai., "S)'nthc.~is a nd Evaluation of So me Wa ler(cid:173)
`Soluble Prodrugs and Derivalives of Taxol wilh Antilumor
`Activity" J. Mcd. Che/ll., 35: 145-15 1 (1992),
`Norto[) et al.,AbSlmclS of I/'<: 2nd NaliOllal Callcl!r /II$lillll<:
`Workshop 011 Taxoi & 'laxus, ScI" 23-24, 1992)
`Wani ct aL "Plant Antitumor Agents. VI. '1lJc Isolation and
`Siruclll rc of 'Ioxel, a Nuvel Antileukemic and AnI;lUmer
`Ag~nts from T(wl~' br;:"jfvlia'·2>, J. Alii, C/II!//l. Soc"
`93:2325-2327 (1971 )
`
`Apotex v. Abraxis - IPR201 8-00151 , Ex. 1026, p.02 of 19
`
`

`

`4000
`
`3500
`
`3000
`.,;-
`::;; 2500
`
`~
`
`::;; -Gl 2000
`E
`'" "0 1500
`>
`0
`E 1000
`'" I-
`
`500
`
`' ; . - -... ------.-----....
`0
`-500lrrrrr
`12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34
`
`Treatment Period
`
`- - - - - - - .... ---- - ...... - -
`
`- - --- ..
`
`Figure 1
`
`Days Postimplant
`
`Apotex v. Abrax is -IPR201 8-00151 , Ex. 1026, p.03 of 19
`
`c::
`
`[Jl
`
`~ ;:
`'" ....
`-c
`-~ -~
`
`?
`
`en
`
`~ a
`Q -....
`
`'" ~ ....
`'" '00
`
`0'\
`
`0'\
`
`

`

`140%
`
`~ • o 120%
`o
`~
`II
`o
`i;'
`'D 100% ~
`.~
`~ 80%
`D.
`~
`~
`~
`~
`
`60%
`
`tJ)
`
`paw Size pre- & PQst- Treatment
`
`Non-treatment
`
`>J
`
`Prednisone 0.2 mglkg +
`
`Pacli!axel Nanoparticles 0.5 mglkg
`
`Paeli!axel Nanoparticles 1.0 mglkg
`
`c
`S"
`~ ...
`'" = ...
`
`c
`C
`
`N :;c -~
`
`~
`
`'J)
`".
`
`&
`N
`o
`~
`N
`
`40%~1------__ -----.------r------r------, __ ----'-----~------__ -----.------.
`Pre-Arthr Day a
`Day 3
`Day 5
`Day 8
`Day 10 Day 12
`Day 20
`Day 25
`Day 32
`Day 49
`Days After Treatment
`
`Figure 2
`
`Apotex v. Abraxis -IPR2018-00151 , Ex. 1026, p.04 of 19
`
`Y'
`\C
`......
`
`0\ '" \C
`
`0\
`
`

`

`5,9 16,596
`
`PROH]N STA8IUZEIl
`PHARMACOI.<>G ICALLY ACTIVE AG ENTS,
`METHODS FOR THE I'REJ"ARATJON
`TlI EREOF A ND METHo n s FOR TIlE USE
`THEREOF
`
`RELATEr) AI'I'I J('A'n ONS
`
`This application is a continuation.in.pan of U.S SUo No
`08/412,726, filed Mar. 29, 1995. now issued as U.S. Pat. No.
`5.560.933, which is, in turn, a divisional of U.S. Ser. No.
`081023,698, fikd r",b. 22, 1993, now issued as U.S. Pat. No.
`5,439,686, the en tiTe contents of both of which are hneby
`incorporated hy rcfw;1"lCC herein in their entirety
`
`FI ELD 01-' T ilE INVENTION
`
`·lhG prescnt inve ntion relates to methods for the produc.
`ti on of panieul.te veh ide ~ for the intravenous administra(cid:173)
`tion of pharmacologically active agents, as well as novel
`compositions produced thcreb~'. In a panicular aspect, th e
`invention relates to methods for the in vivo ddivery of
`substantially water insoluble pharmacologically active
`agents (e,g., the anticancer drug taxol). In another aspect,
`dispersible colloidal s~'Slems oontaining water insoluble
`pharmacologically act iv~ agems ar", provilkd. The sus(cid:173)
`ptonded particles are encased in a polymeric shell formulated
`from a hiocompatible polymer. and have a diameter of lcs,,,
`th an about I micron, Invemion oolloidal systems arc pre(cid:173)
`pared without the use of conventional surfactant or any
`polymeric <-"re matrix. In a pre!;t:lltly prt:fcrre,j aspect of the ] 0
`invention. there is provided a method for preparation of
`extre mely ~a!l particles which can be sterile·fi ltered. The
`polymeric shell cOn1ains panicles of ph3rmacologically
`a<-1ive agent, and opt ionally a biol.'Ompatiblc dispersing
`agent ill whic h pharlllaoolo~cally activc agent can be either 35
`diK..-.nlved Or ~u~pcfl(lcd. Thus, the inve ntion providcs a dm g
`delivery system in either liquid form o r in the from of a
`redispersible powder. Either fonn provides ooth immedi(cid:173)
`ately bioavailablc dru~ molccuks (i.<:" drug mokeules
`which arc molecularly bound to a protein), and pure drug 40
`particles coated wi th a prolein.
`
`2
`orgallS arid tiss u~s allows thc free I",ssage of the!;t: bloocl
`celk When microthrombii (blood clol~) of size greate r than
`10--15 microns are present in circulation, a risk of infarc tion
`or blockage of the ca pillaries rcsults, leading to ischemia o r
`oxygen deprivation and possible tissue death. injection into
`the circuJatiou of particles greater than 10-15 microns i"
`diametcr, therefore, must be avo ided. A su.~pcnsion of par(cid:173)
`ticles less tban 7-8 microns, is how",vcr, rclatively safc and
`has been u~d for the deJiVtTy of pharmacologically acti~
`10 agems in the form of liposomes and emulsions, nutritional
`agems, and comrast media for imaging applications.
`The size of panicles a oo their mode of delivery de ter(cid:173)
`mines their biological behavior. Strand et a l. (in
`Micro:;p/um:s·lJiUlIff!Jiclll Appii"Ulium, eu, A
`R~llll.J~Ulll,
`15 PI' 193-227, eRe I'rcss( 19t!t!) have described the fate o f
`parlicles to be dependent on their ~ir.c . Particles in the sir.c
`range of a few nanometers (nm) to 100 nm enter the
`ly mp hatic capillaries following ime[1;titial injection, and
`phagocytosis lllay ()!,:cur within the lymph uodes. After
`20 intravenouS/int raartc rial injection, particles lc.~s than about 2
`microns will be rapidly clcared from the blood stream by the
`reticuloendothelial system (RES), also koown as the mono(cid:173)
`nuclear phagocyte system (MPS). Panicles largt:r than about
`7 microns will. after intravenous injection, be trapped in the
`25 Inng capillaries. After intraarterial injection. panicles arc
`trapped in the first capillary bed reached. Inha led particles
`an;: tra pped by tht alvcolar macrophages.
`I'harmaceuticals that are water-insoluble or poorly water(cid:173)
`s.olublc and !;t:nsitivt; to acid environmcnts in lbe stQlmch
`cannot be conventionall y administered (e.g .• by intravenOus
`injecti'," or oral a"mini.~tration) The parentera l ad mini...rl1l(cid:173)
`tion of such pharmaceuticals has been achieved by emulsi(cid:173)
`fication of the oil solubilized drug with an aqueous liquid
`(such as nunnal saline) in the prescnce of surfactants or
`emulsion stabilizcrs to produce st.ble microcmulsions
`These emulsions may be injected intravenously. pTm'ided
`the components of the emulsion arc phannaoologically iocrt
`U.s. Pal. No. 4,073,943 deS<.Tibcs the admi rl istrati,)O of
`water-insoluble pharmacologically active age nts d issolved
`in oils and emu lsified with WJter in the presence of s urfac(cid:173)
`ta nlS s uch as egg phosphatides, pluronics (copolyme rs o f
`polypropylene glycol and polyethylene glycol), polyglyc(cid:173)
`erol olcat~ , ~tc, I'cr internatiollai l'ublic~tjon No, WOOS!
`OlIO! I describes pharmaceutical microdroplets of an anaes·
`thetic coated with a pho spholipid such as dimyristoyl
`phospbatidylcholine having suitable dimensions for intrad-
`",noal or imrav~nous injt:ction.
`i\n example of a water-insoluble drug is taxol, a natural
`produd first isolated from the Pacific Yew tree, lilX"~'
`SU br~ifofjlf, by Wani et a1. (J. Am. ellclII. SIX. 93:2325
`(1 971 »). Among the anlimitotic agcn l~, taxol, wh ieh contains
`a diterpene carbon skeleton, exhibits a unique mode of
`action on microtubule proteinS responsible for the fonnation
`of the mitotic s pindk. I" contrast with o ther antimitotic
`5S agems such as vinblastine or colchicine, which prevent the
`assem bly of tubulin, taxol is the o nly plant product known
`to inh ibit the depolymerization IITOCeSS of tubulin, thus
`pr-.:v~lltillg the wll replkation proccs.s.
`TaxoL a naturally occurring diterp.::noid. has been shown
`60 to have significant antineoplastic and anticanct:r dfe"bi ill
`dru g-refractory ovarian cancer. Taxol has shown excellent
`antitumor activi ty in a wide variety of tu mor models such as
`the H!6 melanoma, U210 leukemias. MX-J mammary
`tu mors, and CS-I culon 1l!lllor xcnografbi. Se"eral rc~"'n t
`65 press re leases have termed taxol as the new amicanccr
`wo nd cr.dmg I"'lccd, taxol has recently necn approved by
`the Federal Drug Administration for trea tmen t of ovarian
`
`UACKUROUNIJ 0 1' TI lE INVENT ION
`
`Intravenous dmS delivery permits rapid and direct tquili- ~5
`oration with the blood .~ ream which carries th e medication
`to the rest of the body. To avoid the peak serum levels which
`arc achieved within a shor1tim'" after i01rava!;Cuiar injection.
`adm inistration o f dru~ carried within s table carriers would
`allow grad ual release of the dOl £.-" in~idc thc intravasculM
`companment following a bolus intravenous injection of th e
`the ra peutic nanopa rt icies.
`Injectable controlled_release nanoparticlcs can provide a
`p re-p rogrammed du ration of acti on. ranging from days to
`weeks to months from a single injection_ They also can ofier
`sevtral profound adva01ag~s ovcr C()n\'cntionally adminis(cid:173)
`tered medicalf)ents, including automati" assured patient
`eomlllial"lCC with the Onse regimen, as well as dmg targeting
`to specific tissues or organs (l Ice and Gilley, Journal Of
`C()fJ{rolled Release 2:343-352 (1 985).
`Mieroparticles and fo reign bodies pre.se01 in the blood are
`generally cleared from the circulation by the "blood filtering
`organs" , namely the spleen. lungs and liver. The particulate
`lllal1~r contained in nurlll~l ",hoI<: blood <.:ompriso:s [~d blOO!l
`cdls (typically 8 microns in diameter). whit'" blood cells
`(typically 641 micro ns in diameter), and platelets (typically
`1- 3 microns in diameter). The microcirculation in most
`
`Apotex v. Abrax is - IPR201 8-00ISI , Ex. 1026, p.OS of 19
`
`

`

`5,9 16,596
`
`15
`
`3
`can~"Cr. "Il", poor aq u<X)u~ S<)iubilil y ..,f lax!)l, huw~vcr,
`prese nts J problem for human adminislralion. Indeed, th e
`delivery of drugs that arc inherently insoluble or poorly
`soluble in an aqueous medium can be seriously im paired if
`oral tlc\ivcry is ]IQ I effective. AcL'Ortlingly ... "ur"'''l]y us<:d
`taxo l formula tions rcquuc a crcmaphor \0 sol ubilize th e
`dmS. The hll man clinical ,klsc range i" 200-5fXI mg. This
`dose is dissolved in a 1: 1 solution of elhanol:crcrnaphor an d
`diluted 10 one lite r of Huid given in travenously. The crema(cid:173)
`phor currcmly uS<'o is poly~lhoxyl altll castor uil.
`In ph a<\C [ c1in icall rial ~ laxII1 itself did nOI show exces(cid:173)
`sive loxic c lIecls, but severe allergic reactions were caused
`by lht t mulsifit rs employed 10 solubili~x tho" dru g. The
`currcnl regimen of adm inistration involves trealmenl of th e
`patient wil li antillistam ines and sleroids prinr 10 injeclinn of
`the drug to reduce Ihe allergic side elfectsof ille ercmapllore.
`III all ~If...,rt I...,
`iJJlpr...,v~ th~ waler ~lubi lit y !)f tax!) l,
`several investigatoc:s have mooified i1.s chemical Slnletllre
`with funct ional groups Ihal impa rt enhanced waler(cid:173)
`solu bility. Am ong them arc Ihe sulfonated derivatives
`( Kingston et aI., U.S. Pal. No. 5,059,699 (1991». 3ml ami no
`acid est".." (Mathew "t aI., J. lHcd. ClI<.'111. 35,145-151
`(1992» which .~how s ignifican t hinlogical act iv ily. Modifi(cid:173)
`ca tions to produce a water-soluble derivative facilitate the
`intravenous delive ry of laxo! dissolved in an innocuous
`",arrier s ud, as o...,un al "..Ii"e. Sud, m<.Kiifica ti...,lIs, h...,wever,
`a1kl 10 the eOSI of d rug prep3ration, may induce undesire d
`side·react ions and/or al lergic reaction;., amJJoT may decrease
`the efliciency of tDc d rug.
`Protein micrn:<;phere~ havc heen reporled in Ihe lilcrature
`as ca rriers o f pharmacological OT diagnost ic agents. Micro(cid:173)
`spheres of albumin have been pTepa red by either heat
`denatura tion or c h"mical crossli nking. Ileal denalured
`m iCf"4),;pl>e re.~ are prndllced from .n emulsified m i>:rure (e.g. , 35
`alhumi n, the agenl to he incorporaled, a nd a suitahle oil) at
`tempera tures belween 1000 C. and 150" C. ·llle micro(cid:173)
`sphnes are then wasllt:d with a suitable solvent amI stored.
`Leucula el a1. (I"ternational Jo<,rnal of Phllrmar:elllir:$
`41,213-2 17 (198g» describe th e method of preparation of 40
`heat dena tu red microsphe res.
`The proced u,,; for prtpar ing chemicall y crosslinked
`micrn:<;phe res involves treating the emu lsion wilh glularal(cid:173)
`dehyde to crosslink the protein, follow ed b>' washing and
`s torage. Lee et aJ. (Science 213:233-235 (1981» and U.S.
`Pat. No. 4,671,954 teach th is method of preparation.
`the preparation of protein
`·llIe above techniques fOT
`mil.Tosp heTes as carriers...,f pharmaoo logically acti ve agenls,
`although su itable lor th e delivery of waler·soluble agen ts,
`arc incapahlc of entrapping water_in,,"luhle OneS _ This limi(cid:173)
`tation is inhe rent in the technique of prepaTation which relies
`on cross li nking o r heat denaturation of the protein compo(cid:173)
`nent in Ihe aqueous phase of a water.iLl·oil emulsio n. Any
`aqueoos-soluble agen t disso lved in the protein-containing
`aqueous pllase may he entrapped withi n the resultant
`crosslinked or heat-denatnred protein m3uix, but a poorly
`3'lutous-solublc or oil-solubk asen t canno t I,.; incorpora te d
`into a prolein ma trix lormed by th ese techn iques
`One conventional method for manufacturing drug(cid:173)
`",...,ntai ning naulJl'a rlicles l."{)rU l'riscs d iSS<.llv ing p<Jly laeli ~
`acid (o r other biocom patible, water insol uble polymers) in a
`water·i mmi<;eihle solvenl (such as melhylene chlnride or
`other chlorinated, aliphalic. or aromalic solvent), diss.olving
`the phanna<xu tical1y active asent in th e po ly"'~r solu tion,
`add ing a surfactant to the oil phase or the aqueous phase,
`forming an oil_i n_water cmul"ion hy "nil.hle means, .nti
`evaporating the emulsion slowly under vacuum. If the oil
`
`4
`drop lets arc s ufficien tl y ,,(u all and stable d uring cvap<Jrati...,,,.
`a suspension of Ihe polymer in waler is obta ined. Since the
`drug is init ially prC<;enl in the polyme r solutio n, it is po~sible
`10 oblain by this melhod, a compositio n in which Ihe drug
`molecules arc entrapped w ilhin particles "· .. lInpused ')[ a
`polymeric matrix. The fOflualion of microsphcres and nano (cid:173)
`parliclc, hy u_~ing the solvent e"aporation method ha s heen
`reported by scveral researche.." (see, for e~ampJe . Tice and
`G illey. in Joornal of Corurolk'f1 Rcl(,II~·(! 2:343-352 (1985);
`10 Bodmo: kr and McGinit ~, in lnt. J. Pllllfllll,cew ics 43:179
`(1 988); Cavalier et aI., in J PllI/rlll_ PIJarlllllcol. 38:249
`(1 9R5); and D' Souza el aI., WO 94{I09RO) while using
`various drugs.
`Bazile el. aI., in n;o<""leria/s 13: 1093 ( 1992), ami Spen-
`lehauer et a I. , in Fr Pate nt 2 660 556, have reported the
`formalion of nanopanicles by using two biQCQmpatib1c
`polymers, one (e.g. Polylact ide) is dissolved in the organic
`phase, t!)gclh~r with all active wmpooc nt such ~ a drug, and
`the other polymer, such as a lbumin is used as the surface
`20 active agent. After emulsificalion and remm 'al of the
`solve nt, nanoparticle:s are formed, in wh ich l he drug is
`prt:sell t inside tilt: (X.llymeric: matrix o f tile polyl actitle par(cid:173)
`licks.
`The properties of the polyme r solu tion from which Ihe
`polymeric ma tri x is formed are very important to obtai n Ihe
`proper emulsi...,n ill Ih~ fir.;t "tage. FIJf e;um pk, »IJlyla~tide
`(the polymtr commonly used in lhe preparation of injectable
`nanoparticles), has a surface activity which causes the rapid
`ad-.orption thereof at Ihe die hloromelhane -watcr interface,
`causing redul."\:d interfacial te nsilJ n (se~ . for exampk , llou ry
`et aI., in Laflxmllir II: 1636 (1995 )), which in turn imprO\'es
`lhe emul s ificatio n pmcc ... ~. In addilion, the Mmc researchers
`found that Bovine Serum Albumin (BSA) inte racts wilh the
`polylactidc, and penetrates into the polylactide monolaye r
`prt:"", nt at the oil_water interface. -n'~n:[...,re, it ;,; expected ,
`b",'\Gd o n Ihe above refe re nce. thaI emulsiticalio n during Ihe
`conventi onal solvent evaporation melllod is greal1y favored
`by the presence of tilt: surface active polymer (polylaclide)
`in the nonaqueous organi!.! phase. In facl, the pn:sence o f
`poiylaclide is not o,tly a sufficien l condilion. bU I il is actually
`n cces.~ ry for the formation of naTlOpMticlc.~ of ",,,ilaole ",i~c
`Anotllcr process which is based on the solvenl evapora(cid:173)
`lion mdhod .. "Om priscs di ssolving the drug in a hydrophobic
`~5 solve nt (e _g, tolue ne o r cyclohexane), wilhout an y polyme r
`dis.<;ol\'ed in Ihe organic solvcn t. addi ng a convcntional
`surfaclam 10 the mixture as an emulsifie r, forming an oil(cid:173)
`in-wat.er emulsion , and thtn evaporating the ~lv~m to
`obtain dry parlicles of the drug (see, for example, Sjostrom
`5U et aI , in 1. n i\fJCrsinn Science and Teclmnlngy 15 :1'\9-117
`(1994). Upon removal of the nonpolar solvent, predpit3 tion
`of the dru g inside the solvent droplets occurs. and submicron
`particks arc obta ined.
`II has heen found that the size of toc particles is main!)'
`5S co ntrolled by tile initial s i"(.e of the emulsion droplets. In
`atltl itio n, it is i nt~rt:s1.ing to note Iha ttllt: fin al panicle size i~
`report<:d to decrease wi th a decrease in the dmg concentra_
`tion in the organic pha.'<C. This finding is cont rary to the
`resul ts reported herein, wherein no conventiona l surfac t3 nt
`60 is used for th e preparation of nanoparticles. l n addi!ion, it is
`"'.lIed by the authlJl"l; ...,f Ihe SjlJSlro,,, paper th at the drug
`use d, cholesleryl aceta Ie, is surface active in toluene, and
`hence may be oriented at the oil· water in terface; the refore
`Ihc C(>nCtlnlration of drug al the in tcrface is bighe r, thus
`incTeasing the po tential for precipitation.
`Form ation of snhmicron particles ha", also ocen achievCll
`by a precipita!ion process, as described by Calvo el al. in J.
`
`25
`
`.w
`
`65
`
`Apotex v. Abraxis - IPR20 18-00 15 1, Ex. 1026, p.06 of 19
`
`

`

`5,9 16,596
`
`S
`PlllIrm, 5<.:i. 85:530 (1 996). 'Ill~ VNCCSS is uased l)[l dis.wlv>
`ing the dnl!;. (e.g., indo methacin) and the polymer (poly(cid:173)
`capmlaclonc) in methylene chloride and aoxtonc, and then
`pouring the solution inlo an aqueous phase conlaining a
`s urfal'la!l! (Polo xamcr 18S). to yield submicron si~..o: par.
`ticks (21 6 11m). However, the process is performed at
`sc.tv';nl ooncentrations 31 which no cmlllsion is formed
`
`6
`vivo delivery nf suiJ"lanlially water in>;<.>lubk pharlll ac()l()gi_
`cally active agellis. Invention composilions comprise sub(cid:173)
`slantially water insoluble pharmacologically active agents
`(as a solid or liquid) contained within a polymeri c shell T hc
`polymeric shell is a crosslinked biocompatible polymer. The
`poly meric shell, containing substantially water insoluble
`pharmacologically active agenL~ locrein, can Ihen Ix: SIL"(cid:173)
`p~ndcd iu a bi(X.vmpati blc aqueous liquid ror administra(cid:173)
`tion
`·The. invention further provides a drug delivery syslem in
`which part of Ihe molecules of phar macologically aClive
`agent a re bound to Ihe protein (e.g., human scrum albumin),
`and are thcrefore immediately bioavailable upon adminis(cid:173)
`Iration to a mammal. '[be Ol her portion of the pharmaco-
`15 l()gkally adivt agent is oonlained within na tHJpanidcs
`coated by protein. 'Ibe nanopartides cOlliaini ng the phar(cid:173)
`macologically active age nt arc prc.scnt a.~ a pure active
`componenl, withom dilution by any [lOlymeric matrix.
`A la rge numlx:r of convcmional pharmacologically active
`20 agems circulate in the blood stream bouoo to carrier proteins
`(through hyuwphubic Of ionic inlCrJctio ns) ()f whid Ihe
`most common exam ple is serum d bumin. Invention meth(cid:173)
`ods and compositions produced thereby prnvide for a phar(cid:173)
`macologically active agen t that is "pre-bound'" to a pro lein
`25 (through hydrophobic or io nic interactions) prior to admin(cid:173)
`istratiun.
`The. present disclosure Uemonstra t.:.s both o f the ab()v~­
`described modes of bioavailabilit y (or Taxol (Paclitaxei), an
`amic~ ncc r drug ca pable of hinding to hu man serum alhumin
`.w (see, for example, Kumar el aI., in Xesearcil COIIIIII!Ulica(cid:173)
`rions in Chemical {'mh%gy and {'harmacology 80:337
`(1 993»). 'n", high "on"entratK.Hl ()f albu()liu in inv~ ntiOIl
`parI ides, compared to Taxol, provides a signilicam amoum
`of the drug in Ihe form of molecules bound to alhumin,
`35 which is 3lso the natural carrier of the drug in th e blood
`slream.
`In addition, advantage is taken of the capability of human
`serum albumin to bind Taxol, as w~lI as other drugs, which
`enhances lhe cap.bility of' ["axollo absorb o n the surface of
`Ihe pa niclcs . Since al bumin is pre.<enl nn th e colloida l drug
`parlicl es (formed upon removal of the organ ic solvem),
`formal ion of a colloidal dispersio n which is stable fo r
`prul<.lllgcu po.;riods is ra"ilitated, due to a ,-'Omhination o f
`electrical re pllisio n and steric st. bili/Jltion
`[n accordance with the pft:St;nt invcn tion, thcre arc also
`provid ed submicron pa rticles in powder form, which can
`easily be reconstituted in water or saline. The powder is
`oblained afte r removal of water by lyophilization. Human
`serun, albuluin St:rvcs as the stru"tural ",omponent ()f illve n_
`tion nanoparticies, and also as a cr~oprotec.tam and recon_
`stitulion aid. The preparatio n of particles Ilherablc th ro ugh
`a 0.22 micron filter according to the inventioll method as
`described hercin, followed by drying or lyophili7..ation, pro(cid:173)
`duces a s1crile solid forlllulalion useful for inlravenous
`injection.
`The invention provides, in a particuhr aspect, a compo(cid:173)
`s ition of anli...::ancer dnt gs, e.g., Taxol, in the form o f
`nanopanicles in a liquid dispersion or as a solid which can
`be easily reconst ituted for administration. Due to specific
`pruperti~s ()f ""rtain drugs, c.g., Taxol, Slid, w n11,..,,;itions
`can not be obtained by con\"emional solvent evaporation
`methods that rely on the usc of surfactanl~. In loc presence
`of various surfactants. very large drug cryst3L~ (e.g., si~e o f
`about 5 micwus to scv~ral hundftJ micro ns) are rOfm~d
`65 wilhin a few minules of storage, after the pre paration
`process. The si1.c of .... 'ch cry.~lals is typically much grc.tcr
`Ihan Ihe allowed si7.e for inlravenous injcclion
`
`1U
`
`BRIEF DESCRIPTION OF '11m INVENTION
`"Ibus it is nn object of Ihis invention \0 deliver pharma(cid:173)
`cologically active agents (e.g., taxol, llxane, TaxOlere. an d
`the like) in ulllllodilied form ill a composition that does not
`calise allergic reactions due to the presence of add«1 emul(cid:173)
`sifiers and solubilizing agents. as are currently employed in
`drug delivery.
`11 is 3 further ohjcCl of the prc."Cnt invention 10 deliver
`pharmacologically aCl ive agenlS in a composilion of micro(cid:173)
`panicles or nanopanicles. oplionally suspended in a suilable
`bio<':ompali\.Jk Ii<j uid.
`h is yel another objeel of the presem invention 10 proville
`a mdhod fo r Ihot formal io n of subminon panicles
`(naoopa rtidcs) o f pharmacologically active agen ts by a
`solvent evaporation lechnique from an oil-in-watercmulsion
`using protein:> a:; stabilizing age nts in the absence of any
`conventional surfaetan\S, and in Ihe absence of any poly(cid:173)
`IIIeri" ~vrc mat~ ria l.
`'lbest and o ther objects of th e invention will become
`apparent upoo rev iew of th~ spc~ ifi~a tio n amJ dairru;.
`In accordance w il h Ihe presenl invenlion. we have dis(cid:173)
`covered Iha t subStanlially waler insoluble pharmaoologi(cid:173)
`"aliy a"tiv" agents "an b-c ddivucd in thc rurlll of ,nino(cid:173)
`particles or nanopanicles lhal are suilable for parenleral
`admin istr& lion in aq ueous suspension. This mode of delivery
`obviates the necessity for adm inistrat ion of substantially
`water insoluble pharma,-vlogically acti~ agents (e.g .. taxol)
`in an emulsion COtllaining, for example, elhanol and poly(cid:173)
`elho~yla ted Ca$lor oil, dilnted in normal saline (see, for
`eump1c. Norton el aI., in lIbsrracrs of rlie 2nd Nmiona/
`Cancer Insrirwe Worksllop on Taxol & Taxl/s, ScI" 23- 24. 40
`1992). A disadvantage "f su"h known COlllp()sitions is Iheir
`propensity 10 pro<luce allergic side c!fccts
`nlU~ in a"oordam.-": with the prcso.:ut iuventiuu, tll",rc ar~
`provided methods for the formation of n.nop.rticies of
`pharmacologically lICtive agents by a :;(llvent evaporation ~5
`technique from an oil