United States Patent [19]
`Li'fersidge et aI.
`
`[54) SURFACE MODIFIED ANTICANCER
`NA..'10PARTICLES
`
`[75]
`
`Invenlon: Gu)' G. Unrsidge; Elame
`UTel'Sidr;e. both of West Chester;
`Pramod P. Sarpotdar, Malvern, all of
`p~
`
`[73) Assignee: EuI:DIan Kodak Compauy,
`Rochester, N.Y.
`
`[21]
`
`Appl. No.: 908,125
`
`[22]
`
`Filed:
`
`Jul. I, 1992
`
`IIIIII IIIII~IIIIII~ 111111111111111111111111111111 .lllIIm 1111111111
`USOO5399363A
`5,399,363
`[II] PateDt Number:
`[45] Date of Patent: Mar. 21, 1995
`
`FOREIGN PATENT DOCUMENTS
`262!160 9/ 1911 &ropan Pal. On: .
`411629 21 1991 ~PlI. on: .
`499299 1/ 1992 Ellropean Pal. OfT . .
`2118981 4/ 1972 France .
`3n2831 1/ 1987 Germany .
`2282330 11/ 1990 JapIUI .
`21U397 1/ 1987 Uniled Kingdom .
`22<XXM8 7/1988 United Killgdom .
`8400294 7/1983 WlPO .
`91 1$193 6/1989 WIPO .
`901$593 6/1990 WIPO .
`9203380 8/1990 WIPO .
`9106292 11/ 1990 WIPO .
`
`Related U.s. ApplicadoD Data
`[63J Continualion-in_part orSer. No. 647,]05, Jan. 25, 1991,
`Pac. No. '. 14MM.
`fat. 0.6 ......................... _ .................... A61X 9/14
`[5 1]
`[52) U.s. a . ............. _ ...................... 424/ 490; 424/0489
`[58) Field of Seartb ._ .............. 424/ 490, 487; 514/352
`
`(56)
`
`Rereruce:s Qed
`U.S. PATENT DOCUMENTS
`.. 514/179
`2.671,750 1/ 19S4 M.acc: .... .... _ ... __ ._ .... _ ...
`l .lIl,QlO 4/ 197S Nakamura et aI. _ .. _ .. _ ... _. 514/ 619
`4,107.2&8 1/191& Oppenheim _____ " _,,_,,_,,, 424/22
`4,m,SS I ~1I980 Kreuter ct aL __ .. __ _ .. _ ...... _ 424/18
`4,269,321 5/ 1911 Krellttl' eI aI. ~ ~ ~ •• _ .. ~~_ •• 424/ 419
`4,540,602 9/ 198$ MOIoyam;o ._ ._ .. __ .. _ 421/213.3 1
`4,S26,619 $/ 1989 Voolanto .. _ .. _ ... _ ...... _ .. _ .... 424/489
`4,1$1,421 7/19&9 Iwasaki el aI. •. _._._ .. _ .. _ ~ 14/3~2
`$,~9,322 9/ 1991 DeviMaguct _ ~_ .. _ .. _ .. _ .. _. 424/490
`$,091,188 211992 Hayoes ... _ .. _ .. _ •..... _ .. _ .. _ 424/4SO
`$,I II,S2S 6/1992 Fessi ........ ~ ...... _ •. _ ... _ ....... ~ 424/481
`5,124,338 6/1992 Kin, ........ _ .. _ ....... _ ...... _ ... _ 514/3$2
`
`OTHER PUBLICATIONS
`Lachman et aL, ''TIle Theory and P ractice ofIndustria1
`Pharmacy", Chapler 2 (I986).
`RemingtOn's Pbannaceutical Sciences, 17th Edition,
`Chaplet 20, Schou, H., "Colloidal Dispersions".
`Goodman and G ilman, "The Pharmacological Basis of
`Therapeutics", E ighlh Edition, pp. 68-69.
`Primary Ex.amintr-Thurman K.. Page
`Assistant Examiner-William E. Benston, Jr.
`Attortlt}l, Agtnl. or Fjnn-Arthur H. Rosenstein
`[57]
`ABSTRACf
`Dispersible particles consisting essentially of a CTysllll·
`line anticancer agent having a Rltface wodUIICT ad(cid:173)
`sorbed on the surface thereof in an amount sufficient to
`maintain an effective average partick size of k:ss than
`about 1000 nm. Anticancer compositions comprising
`tbe particles exh.ibit reduced toxicityand/or e nhanced
`efficacy, and can be administered by lV bolus injection.
`
`17 Oaims, No Drswings
`
`Apotex v. Abraxis - IPR2018-00153, Ex. ]005, p.O I of 10
`
`

`

`1
`
`5,399,363
`
`SURFACE MODIFIED ANTICAJ'Ii'CER
`NANOPARnCLES
`
`CROSS REFERENCED TO RELATED
`APPLICATION
`This application is a conrinuation.in.part of U.S. pa(cid:173)
`lent a pplication Ser. No. 647,lOS, filed l an. 25, 1991.
`now U.S. Pat. No. S, 145,684, the d isclosure of which is 10
`hereby incorporated by reference in its entirely.
`
`2
`h i, a no ther IIdvtlnto.geous fell.lure of this invention
`that anticancer compositions are provided wibiting
`improved effICaCY.
`Yet another advantageous feature ofthis invention is
`that compositions are provided featuring poorly soluble
`anticancer tgenl$ Ihal can be administered by IV bolus
`inje<:tion.
`Still another advllAtageous feature OhM invention is
`that compositions are provided containing poorly solu(cid:173)
`ble anticancer agents exhibiling prolonged circulation
`in the blood pool after IV bolus injection.
`Other advantageous features will become readily
`BACKG ROUND OF TIlE INVENTION
`apparent upon reference to the folJowing descriptions
`I. Field of Invention
`of preferred embodiments.
`This inventio n relates to anticancer a gents in the form ]5
`DESCRIPTION OF PREFERRED
`of particles, to anticancer compositions comprising the
`EMBODIMENTS
`particles, and to methods employing thc particles.
`2. Descriptio". o~ the ~ An
`This invention is based partly on the discovery that
`.
`Th7 therapeutl~ md~x IS a .measure of how selective a
`surface modified anticaDct:T nanoparticles exhibit ce-
`drug IS at p~ucmg Its d~tred effeclS and can be ~ 20 duced to~ity and/oT enhanced efficacy. While the
`fined ~ the ralio. o(the medwliethal dose to the median
`invention is described herein primarily in connection
`e~ectl.ve dose, I.e., (LD~~) (~ Goodman ~d with its preferred class of drugs, i.e., anticancer agents
`G.ilmaD, .1"!" l'1IarmacolO8.u:tl1 Bosis of. TheraptUtlcs,
`including immunosuppressive agents, it is also useful in
`Eight Edition, p. 68~): Virtually a1l antieancer agents
`conjunction with poorly water soluble drugs. particu_
`have a .'ow therapeutte ~.ex, e.g., less than ~ut 1.0. 2S lady tbose with low therapeutic indices, from other
`~~easmg the .therapeutic mde.l, e.g., ?y reductn~ tox-
`classes of drug substances.
`»elty or ~~g ef.licacy would p~vid~ morc ~tltude
`The particles ohills invention comprise an anticancer
`to ph)'SICI~ In ~etr duty of administenng antJCanCer
`agent The anticancer agent is present in one or more
`drup to patJe!lU ~. need thereof. Consequently, ~.
`discrete crystaJline phase$. Tbe crystalline phase differs
`ocis to red,," toxiCIty ~or cnhan~ efficac.y ,?f -:ntl.- 30 from an amorphous, i.t., DOft-cr)'$tllline phase which
`results from conventional solvent prccipillltion lech-
`cancer drugs and thus increase the t~bc mdJCeS
`niques for the preparation of partic.Ic:s in the sublIlia"OJI
`of such drugs would be of great value In the treatment
`or ~.
`size range, such 15 described in U.s. Pat. No. 4,g26,689.
`In addition, poorl y water-soluble drugs. such as
`The invention can be pncticed with a wide variety of
`~r1y wat~-IOh ... ble anticancer agents, ~ no~ readily lS anticancer Igents. However, the anticancer agent must
`mJCC~le ~ ~ mtnavenous (IV) bolus lDJCCUOn. 1be
`be poorly soluble and dispersible in at least one liquid
`creatiOn of In.JCC~ble forms of poorly soluble ~rug5 medium. By "poorly soluble", it is meant that the drug
`represents I formIdable problem. It would be highly
`substance has a solubility in t he liquKi dispcrsiOJl IDC-
`desirable to be able to p~vide poorly ~I uble drugs, 40 dium, e.I., water, of less than about 10 mg/ml, and
`such as poorly soluble anocancer agents, lD an IV bolus
`preferably. of less than I mglml at processing t~pera-
`injectable form.
`ture, e.g., room temperature. The preferred liquid dis-
`SUMMAXY OF THE INVENTION
`pef$ion medium is water. However, the invention can
`be practiced with other liquid media in which the anti-
`We have discovered thJl.t anticancer compositions ,., cancer agent is dispenible includiDg, for e1lllTlple, aque-
`comprising anticancer agents in the form of surface
`ous sait solutions, saffiower oil, and solvents such as
`modified nanopartieles exhibit reduced to~city and/or
`ethanol, t-butanol, henne, and glycol. The pH of the
`enhanced efflClCY·
`aqueous dispenion media can be adjusted by techniques
`More particularly, in accordance with this invention,
`known in the art.
`there are provided partieles consisting essentially of a SO
`The anticancer agent prderabiy is selected from al-
`crystalline anticancer agent having a surface modifier
`kylating agents, antimetabolite$, natural products, hor-
`adsorbed on the surface thereor in an amount slliT»eient
`manes and antagonists, and miscellaneous agents, such
`to maintain l1li effective average particle me of lc:ss than
`as radiosensiti%ers.
`about 1000 IUD...
`Examples of alkylating agenll! include aikyiating
`This mvention runher provides an anticancer compo- ss agents bavin, the bis-(2-chloroethyl)-amine group such
`silion comprising the above-described. particles.
`as, for e:umple, chlonnethine, chJorambucilc, mdpha-
`In another embodiment or the invention. there is
`Ian, uramustine, mannomustine, e.ltramustinephoshate.
`provided a method of treating a mammal comprising
`mechlore-lhamiM~de, cyclophosphamide, ifosfamide.
`administering to the mammal the aoove-dc:scn"bed anti-
`and trifosfunide;
`cancer composition.
`alkylaring .. g~llts having a substituted aziridine group
`In yet another embodimc:flt of the invention, there is
`such as, for example, lretamine, thiotepa. triaziquone
`provided a method or enhancing the effICaCY andlO!"
`and mitomycine;
`reducing the toxicity of l1li anticancer agent which in-
`alkylating agents of the alkyl sulfoDlte type. such as,
`for example, busulfan, piposuJrllll, and piposulfam;
`eludes the step of adm..inUtering the agent in the form of
`alk ylating N-alkyl-N-nilrosolU"e8 derivatives, such as,
`lhe abovc:-described particles.
`It is In advant.agcous feature of this invention that
`for eJliaIDple, carmustine, lomustine, scmustine, or strep-
`anticancer compositions are provided exhibiting Ie-
`tozotocine; and alkylating agents of the mitobronitole,
`duced. toxicity.
`ciacarbazine and procarbazine type.
`
`60
`
`6S
`
`Apotex v. Abraxis - IPR201 8-00 153, Ex. 1005, p.02 of 10
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`

`

`5,399,363
`
`3
`4
`Examples of antimclabolites includ1: folic add ana-
`benzalkonium chloride, calcium stearate, glyceryl mon-
`ostearate, cctostearyl alcohol, cetomacrogol emulsify-
`logs, such as, for example, methotrexate;
`ing W8;\, sorbitan esters, polyoxycthylene alkyl ethers,
`pyrimidine analogs such as, for example, fluorouracil,
`e.g., macrogol ethers such as cetomacrogoI 1000, poly-
`fiol.uridine,
`fegafur, cywabine,
`idoxuridine, and
`oxyethylene castor oil derivatives, polyoxycthylene
`flucytosine; and
`purine derivatives such as, for e,lIi3mple, mercaptopu-
`soroitan fatty acid esters, c.g., the commercially avaiI-
`able Tweens TM. polyethylene glycols, polyoxyethyl-
`rine, thioguanine, azathioprine, tiamiprine, vidarabine,
`ene stearates, colloidal silicon dioxide, phosphates, so-
`pentostalin, and puromycine.
`dium dodecylsulfate. carOOxymethylcellulOR calcium,
`Examples of natural products include vinca alkaloids,
`10 carOOxymethyk:ellulose sodium, methylcellulose, hy·
`such as, for example, vinblastine and vincristine;
`epipodophylotoxins, such as, for elWtlple, etoposide
`droxyethylcelluiose, hydroxypropylcellulosc, hydroxy-
`propylmethylcellu1ose phthalate, noncrysta1line cdlu-
`and teniposide;
`lose, magnesium aluminum. silicate, triethanolamine,
`antibiotics, such as, for example, adriarnycine, daunu-
`mycine, docrinomycin, daunorubicin, doxorubicin, mi-
`polyvinyl alcohol (PVA), and polyvinylpynolidone
`IS (PVP). Most of these excip:ic:nts aTe described in detail
`thramycin, bleomycin, and mitomycin;
`enzymes, such as. for example, L-asparaginase;
`in the Handbook of Pharmaceutical Excipienu. published
`joint1y by the American Pharmaceutical Association
`biological response modifiers, such as, for example,
`and TIle Phannaceutical Society of Great Britain, the
`a-interferon;
`Pharmaceutical PrCSll, 1986. The surface modifien are
`camptothecin;
`20 commercially available andlor can be prepared by tech-
`taxol; and
`niques known in the art. Two or more surface modifiers
`retinoids, such as retmoic acid.
`can be used in combination.
`Examples of hOODOnes and antagonists include adre-
`Docorticosteroids, such as, for example, prednisone;
`Particularly preferred surface modifien include poly-
`p£Ogestins, such as, for exam ple, h)!d£Oxyp£Ogeste r-
`vinylpyrrolidone, tyloxapol, poluomers, such as Pluro-
`one caproate, medroxyprogesterone acetate and meges· 2S nic TM F68, FIOS and F127, which are block copoly-
`t£Ol acetate;
`men of ethylene oxide and propylene oxide available
`eslrogens, such as. for example, diethylstilbestrol and
`from BASF, and poloxamines, such as TetroDic TM 908
`(T9Oi), which is a tetrafunctional block copolymer
`ethinyl estradiol;
`antiestrogerus, such as, for exam ple, tamo:tifen;
`derived from sequential addition of e thylene oxide and
`androgens, such :as, for example. testosterone propio- 30 propylene oxide 10 elhyleneOiunine available from
`nate and fluoxymesteronc:;
`BASF, de xtrnn,
`lecithin, Aerosol OT TM
`(A01),
`antiandrogell$, such as, for example. nutamide;
`which is a dioctyl ester of sodium sulfosuccinic acid.
`and sonadotropin-releasing hormone analogs, such
`available from American Cyanamid, Duponol TM P,
`as, for eMmple Jeuprolide.
`which is a sodium lauryl sulfate, available from DuPont,
`Examplt!l of miscellaneous agents include radio5en- 35 Triton TM X -200, which is an :ukyl aryl polyether
`sulfonate, available from Rohm and Haas, Tween 20,
`sitizen, such as, for e:umple, 1,2,4-benZOlriazin-3-amine
`1,4-diox\de (SR 4SS9) and 1,2,4-benzotriuine-7-amine
`40,60 and SO, whic h are polyoxyethylene sorbitan fatty
`1,4-dioxide (WIN 59(75);
`acid es~ available from ICI Speciality Chemicals,
`platinum coordination complexes such as cisplatin
`Span 20, 40, 60 and SO, which are sorbitan esters of fatty
`40 acids, ArW:e1 20, 40, 60 and SO, which are sorbitan
`and carboplatin;
`anthracenediones, such as, for example, mitoXlUl_
`esten of fany acids, available from Hercules, Inc., Car·
`trone;
`bowax TM ]550 and 934, which are polyethylene gly-
`substituted w-taS , such as, for exampJe, hydroxyurea;
`cols available from Union Cubide, Crodc:sta TM F-IIO,
`and adrenocortical suppressants, such as, for CJlam-
`which is a mixture of sucrose stearate and sucrose dis-
`pIe, mitotane and aminoglutethimide.
`4S teacate, available from Croda. Inc., Crodesta SL-40,
`which is available from Croda, Inc., hexyldecyl tri-
`In addition, the anticancer agent can be an immuno-
`suppressive drug, such as, for example, cyclosporioe,
`methyl ammonium chloride (CfAC), bovine serum
`albumin and SA9OHCO, which is CuHn CH2 (CON
`azathioprine, sulfasalazine, methoxsalen and tbalido-
`mide.
`(CH3,) CH2 (CHOH).CH20Hh. Surface modifiers
`The anticancer asents II$Cful in the practice of this so
`which have been found to be particularly II$Cflll include
`invention are known compounds and/or can be pre(cid:173)
`polyvinylpyrrolidone, Pluronic F-I08, polyvinyl alc0-
`pared by techniques known in the art.
`hol and gum acacia
`The anticancer agent can be used alone or in combi(cid:173)
`1be surface modifier is adsorbed on tbe surface o rthe
`nation with one or more anticancer agents.
`anticancer agent in an amount sufficient 10 maintain an
`The particles of this invention contain an anticancer ss
`effective average panicle site of less than about 1000
`agent as described above having a surface modifter
`run. The surface modifier does not c hemically react
`adsorbed on tbe surface thereof. Useful surface modifi(cid:173)
`with the anticancer agent or itself. Furthermore, the
`en are believed to include those which physically ad·
`individu.ally adsorbed molecules of the surface modifier
`here to the surfaee of the anticancer agent but do DOt
`are essentially free of intermolecular crosslinkages.
`As used herein, particle size refen 10 a number aver(cid:173)
`chemically bond to the anticancer agent.
`age particle size as measured by conventional particle
`Suitable surface modiflCTS can preferably be selected
`size measuring techniques w ell known to those skilled
`from known organic and inorganic pharmaceutical ex·
`cipients. Such excipients include various polymers, low
`in the an, such as sedimentation field flow fractionation,
`molecular weight oligomer.;, IIlltUral products and SIll(cid:173)
`photon correlation speclrO$COpy, or disk centrifugation.
`By "an effective average particle size of iCSll than about
`factants. Preferred surface modifien include nonionic 65
`1(0) nm" it is meant that at least 90% of the particles
`and anionic surfaclants. Representative examples of
`cxcipients include gelatin, casein, lecithin (phospha(cid:173)
`have a number average panicle size of less than about
`tides), gum acacia, cholesterol, tragacanth, stearic acid,
`1000 nm when measured by the above·noted tech·
`
`60
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`6
`5
`The mechanical means applied to reduee the particle
`niques. In particularly preferred embodiments of the
`size of the anticancer agent conveniently can take the
`invention, the effective average particle size is less than
`fonn of a dispersion mill. Suitable dispersion mills in(cid:173)
`about 400 run. In some embodiments of tln: invention,
`clude a ball mill, an attritor mill, a vibratory mill, a
`the effective average particle size is less than about 300
`planetary mill, media mills such as a sand mill and a
`nm. With reference to the effective average particle
`bead mill. A media mill is preferred due to the relatively
`size, it is preferTed tbat alieast 95% and, more prefera(cid:173)
`shoner milling time required to provide the intended
`bly, at least 99% of the particles have a particle size of
`resuh, i.e., the desired reduction in particle size. For
`less than the effective average, e.g., 1000 nm.ln particu(cid:173)
`media milling. the apparent viscosity of the premix
`larly preferred embodiments, essentially an orlhe parti(cid:173)
`10 preferably is from about 100 to about 1(0) centipoise.
`cles have a size less than UXlO nm.
`For ball milling, the apparent viscosity of the premix
`Motoyama et aI, U.S. Pat. No. 4,S40,602 disclose that
`preferably is from about I up to about 100 centipoise.
`a solid drug can be pulverized in an aqueous solution of
`Such ranges tend to afford an optimal balance between
`a water-soluble high molecular substance, and that as a
`elI"Jcient particle fragmentation and media erOllion.
`result of such wet grinding, the drug is formed into
`The grinding media for the particle size reduction
`finely divided particles ranging from O.S JUIl or less 10 5 15
`step can be selected from rigid media preferably spheri(cid:173)
`f'l'lI in diameter. However. there is no suggestion that
`cal or particulate in form having an average size less
`particles having an average particle size of less than
`than about 3 mm and, more preferably, less than about
`about IIJm can be obtained. Attempts to reproduce tbe
`I rom. Such media desirably can provide the particles of
`wet grinding procedures described by Motoyama et a1
`the invention with shaner processing times and impart
`resulted in particles having an average particle size of 20
`less wear to the milling equipment. The selection of
`mucb greater than 1 JLDl.
`material for the grinding media is not belicved to be
`The particles of this invention can be prepared by a
`critical. However, zirconium oltide, such as 95% zrO
`method comprising the steps of dispersing an anticancer
`stabilized with magnesia, tirconium silicate, and gJass
`agent in a liquid dispersion medium and applying me- 2j
`grinding media provide panicles having levels of con(cid:173)
`chanical means in the presence of grinding media to
`tamination which are believed to be acceptable for the
`reUuce the particle size of the anticancer agent to an
`preparation of pbarmaceutical compositions. Funher,
`effective average particle size of less than about 1000
`other media, such as stainless steel, litania, alumina, and
`nm.. The particles can be reduced in siu in the presence
`95% zrO stabilized with yttrium, are expected to be
`ofa surface modifier. Alternatively, the panicles can be 30
`useful. Preferred media have a density greater than
`contacted with a surface modifier after attrition.
`about 2.5 g/cml.
`A general procedure for preparing the panicles of
`The attrition time can vary widely and depends pri-
`this invention is set forth bel.ow. The anticancer agent
`marily upon the ,particular medwJtical means and pro(cid:173)
`selected is obtained commercially and/or prepared by
`cessing conditions selected. For ball mills, processing
`techniques known in the art in a conventional coarse 35
`times of up to five days or longer may be required. On
`form. It is preferred, but not essential. that the particle
`the other hand, processing times of less than I day (resi-
`size of the coarse anticancer agent selected be less than
`dence times of one minute up to several hours) have
`about 100 ;.tm as determined by sieve analysis. If the
`provided the desired re!iults using a high shear media
`coarse particle size of the anticancer agent is greater
`mill.
`than about 100 ;.tm, then it is preferred that the particles 40
`The particles must be reduced in size at a temperature
`of the IIlIticancer agent be reduced in size to less than
`which does nOl significantly degrade the anticancer
`100 Jtm using a conventional milling method such as
`agent. Processing temperatures of less than about
`airjet or fragmentation milling.
`30"-40" C. are ordinarily preferred. If desired, the pro(cid:173)
`The coarse anticancer agent selected can then be
`~ing equipment can be cooled with conventional
`added to a liquid medium in which it is essentially insol- 45
`cooling equipment. The method is conveniently carried
`uble to form a premiJt. The concentration of the antican(cid:173)
`out under conditions of ambient temperature and al
`cer agent in the l.iquid medium can VMy from about
`processing pressures which Me safe and effective for the
`0.1--60% and preferably is from 5--30% (w/w). It is
`mining process. For example, ambient processing pres(cid:173)
`preferred, but not essential. that the surface modifier be
`suresare typical of ball mills, attritor mills and vibratory
`present in the premix. The concentration of the surface ~
`mills. Processing pressures up to about 20 psi (1.4
`modifier can vary from about 0.1 to about 90% and
`kg/cml) Me typical of media milling.
`preferably is 1-75%, more preferably 20-60%, by
`The surface modifier, if it was not present in the
`weight based on tbe total combined weight of the drug
`prem.u., must be added to the dispersion after attrition in
`substana:: and surface modifier. The apparent viscosity
`an amount as described for the prem.u. above. Thereaf(cid:173)
`of the prerni.x suspension is preferablY less than about 55
`ter, the dispersion can be miJ;oo, e.g., by shaking vigor(cid:173)
`1(0) centipoise.
`ously. Optionally, the dispersion can be subjected to a
`The premix can be used directly by subjecting it to
`sonication step, e.g., using an ultrasonic power supply.
`mechanical means to reduce the average panicle size in
`For example, the dispersion can be subjected to ultra(cid:173)
`the dispersion to less than 1000 run. It is preferred that
`sonic energy having a frequency of 20-SO kHz for a
`the premU. be used directly wheJ:I II. ball mill is used for 60
`time of about I to 120 seconds.
`attrition. Alternatively, the anticancer agent and, op(cid:173)
`The relative amount of the anticancer agent and sur(cid:173)
`tionally, the surfai;e modifier, can be dispersed in the
`face modifier can vary widely and the optimal amount
`liquid medium using suitable agitation, e.g., a roller mill
`of the surface modifier can depend, for example, upon
`or a Cowles type mUer, until a homogeneous dispersion
`tbe particular anticancer agent and surface modifier
`selected, the critical micd1e concc::ntr"oltion of the sur(cid:173)
`is observed in which there Me no large aggiomeratc8 65
`face modifier if it forms miceUes, the surface area ofthe
`visJ."ble to the naked eye. It is preferred that the prCIDU
`anticancer agent, elc. The surface modifier preferably is
`be subjected to such a prcmilling dispersion step when a
`recin;:ulating media mill is used for attrition.
`present in lID amount of about 0.1-10 mg pet square
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`oxide
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`8
`The selected dosage can be readily determined by oDe
`skilled in the art and depends upon the particular anti(cid:173)
`cancer agent, the desired therapeutic effect, the route of
`administration, the desired duration of treatment and
`other factors.
`It is a particularly advantageous feature that the anti(cid:173)
`cancer compo:sition.~ of this invention exhibit reduced
`toxicity and/or enhanced e fficacy as illustrated in the
`examples that follow. Further, the particles of this in(cid:173)
`vention exhibit prolonged circulation in the blood pool.
`More(lvc:r, anticancer agents which heretofore could
`not be: administered by injection, when prepared as
`nanoparticles and fonnulated in anticancer composi(cid:173)
`IS tions according to this invention, can be effectively
`administered by injection, e.g., by an intravenous bolus
`injection.
`The following examples further illustrate the inven(cid:173)
`tion.
`
`7
`meter surface area of the anticancer agent. The surface
`modifier can be present in an amount of 0.\ - 90%, pref(cid:173)
`erably 0.5-80% and morc preferably 1-60% by weight
`based on the total weight of the dry particle.
`A simple screening process has been developed
`whereby compatible surface modifiers and anticancer
`agents can be selected which provide stable dispersions
`of the desired particles. Fint, coarse p<U1icles of an
`anticancer agent are dispersed in a liquid in which the
`anticancer agent is essentially insoluble, c.g., water al 10
`2% (w/v) and milled for 120 hours in II. rollct mill under
`the following milling conditions:
`Grinding vessel: 8 O~ (250 ml) glass jar
`Available volulIK of grinding vessel: 250 rol
`Media volume: 120 mI
`Media type: 1.0 mm pre-cleaned zirconium
`beads (distributed by Zirooa, Inc.)
`Milling time: 120 hours
`Slurry volume: 60 mI
`RPM: 92
`Room Temperature
`EXAMPLE I
`The slurry is separated from the milling media by
`conventional means, e.g., by pouring the slurry out of
`Pipo$ulfan (purchased from Eastman Kodak) was
`milled in a mixture of 0.33% polyoxyethylene sorbitan
`the vessel, or by using a pipette. The separated slurry is
`then divided into a1iquou aDd surface modifICrs are 25 monooleate, Tween SO, (lCI Americas, Inc., Wilming-
`added at a concentration of between 2 and SO% by
`ton, Del.) and 0.67% sorbitan monooleate, Span So,
`weight based on the total combined weight of the anti-
`(lCI) using I mm zirconium oxide beads for about 96
`cancer agent and surface modifier. The dispersions are
`then sonicated (I minute, 20 kHz) or vortexed using it
`hours 10 produce panicles approxillllltely 240 run in
`multitubro vortexer for one minute, to disperse agglom- 30 diameter. The final piposulfan CQnce:ntr.o.tion in the sus-
`erates and subjected to panicle size analysis., e.g., by
`pen5;on was 10 mg/mL. The panicles were stable to
`photon correlation spectroscopy (PeS) and/or by ex-
`flocculation/aggregation in rat plasma.
`amination under an optical miCIOSCOpe (IOOOXmagnifi-
`Milling Conditions: A coarse: suspension of piposul-
`cation). If a stable dispersion is observed, then the pro-
`fan was prepared by adding 300 mg of the drug to a 4
`celiS for preparing the paniclllar anticancer agent sur- 35 01. (120 mL) amber bottle which was previously filled
`face: modifier combination can be optimued in &C<:or-
`with 60 mL of 1 mm preclc:aned zirconium oxide beads
`dance with the: tc:a<::hings above. By stable il is meant
`(Zircoa Inc., Solon, Ohio) and 30 mL of I % Tween
`that the dispersion exhibits no flocculation or particle
`SO/Span 80 (I to 2 ratio) solution. The surfactant solu-
`agglomeration visible to the naked eye and, preferably,
`tion was prepared by accurately weighing 333 mg of
`when viewed under the optical microscope: at lOOOx, at 40 Tween 80 and 667 mg of Span 80 in a volumetric flask
`least 15 minutes, and preferably, at least two days or
`followed by addition of sterile water for injection to
`longer after preparation. In addition, preferred panicles
`dissolve/disperse the surfactants. Sufficient quantity of
`~bit DO flocculation or agglomeration when dis-
`water was added to make the fmal volume 100 mL.
`pe:rsed in O.IN Hel and/or phosphate butTered saline,
`Zirconium oxide beads were cleaned by fIrSt rinsing in
`pH 7.4 (PBS) or rat plasma.
`45 IN sulfuric acid followed by several rinses with deion-
`The resulting dispersion is stable and consists of the
`ued water. The media was dried in a vacuum oven at
`liquid dispersion medium and the above-dc:scribed pani-
`about 100- C. ovemighL
`c\es. 11Jc: dispenion of surface modifted anticancer
`The sealed primary container was loaded into a sec-
`agent nanopanicles can be spray coated onto sugar
`ondary padded aluminium containment can to ensure a
`spheres or onto II. pharmaceutical excipient in a fluid- SO tight fit. It was milled on a roller mill (US Stoneware:,
`bed spray coater by techniques wcl1 known in the art.
`Mawah, N.J.) at 144 RPM for about 96 hours. At the
`Anticancer pharmaceutica1 compositions according
`end of the milling time the slurry was separated from
`the: media and particle size was measured using a PCS
`to this invention include the particles described above
`and a phaonaceuticaJly acceptable carrier therefor.
`Suitable: phannaeeutically acceptable carnc:n are well 55 device. Stability of these particles to rat plasma was
`known to those skilled in the art. "These include non-
`asscs.sed by optical microscopy al lOOOXmagniflCation.
`The final pH of th.e formulation was 6.
`toxic physiologically acceptable carriers, adjuvants or
`.
`..
`Control A ("l1:mdlc:d). a c~ sus~lOn con~mng
`vehicles for parenteOtl injection, for oral administr.o.tion
`40 mg of bulk plposu1fan was dISpersed III wa~er III the
`in iIOlid or liquid fonn, for rectal adrninistr.o.tion, nasal
`admirristration intramuscular administration subcuta- 60 presence of3% ethanol and 1% Twc:c:n SO. This suspen-
`neous ad~tion, and the like.
`sion could not be injected IV.
`'
`A IIlCthod of treating a IIl&IDmal in accordance witb
`Example 1 was evaluated for efficacy studies in fe-
`this invention comprises the: step of administ.r;ring to the
`male mice: (avg. wI. 22 g) which were implanted with
`early stage Mammary Adenocarcinoma #- 16/C on day
`mamma1 in need oftreatmc:nt an dfective amount of the
`above-dc:scribed anticancer composition. The sc:Iected 65 O."1llc: formulation was injected starting from day I for
`several days. The antitumor activity Wa5 a55CSSed by
`dosage levcl of the anticancer agent for treatment is
`effective to obtain a desired therapeutic response for a
`monitoring tumor weight and comparing it to the con-
`particular composition and method of administration.
`trol animals. The results were as follows:
`
`20
`
`EXAMPLES 1-4 Nanoparticulate Pipo$ulfan
`
`Apotex v. Abraxis - IPR2018-00153 , Ex. 1005, p.05 of 10
`
`

`

`9
`
`5,399,363
`
`T.ca1ment
`
`Control
`Eumple I
`(14lnm)
`
`Route
`T~'
`of Adm. ' Dose (mglks)
`
`'rO W 1.
`
`+5.S
`
`,~
`
`no
`
`.... "
`
`Cell Kill
`
`%
`
`2.75
`1.7S
`US
`
`10
`EXAMPLE 7
`eo. "e
`Blampie S was repeated except that Tetronic 908 was
`replaced by gum acacia. The rwa! particle size was 298
`_ S. 5 ,
`, =.
`_ 5.S ,
`'"
`Nanocamplothecin formulations were evaluated for
`_ 1.B ,
`'"
`efficacy in two murine tumor models, i.e., Mammary
`,
`,.
`'"
`>V
`,y
`Adenocarcinoma # 16/C and Pancreatic Ductal Adeno(cid:173)
`_10.8 ,
`,.,
`"
`"
`carcinoma #03. The antitumor activity was assessed by
`800
`Control A
`SC
`~===-::",==.,.,.;::,:-,--.,-:;';;:;:'-''---'''- ]0 monitoring tumor we .. ' hI from ex ..... rimental and cou-
`. "" ...... """"'. IV In',,",,- sc So.hc._
`,.-
`E_pIe 1 """,1<1 be inJ<ct«ldiuc<lY " 10"'310>1 .... ~ ThOfC""" "" "".It
`trol animals.
`'o>.idly . ft .. in,;....'" <I" mVka "oDe d.,..
`I . Efficacy Studies in Pancreatic Ductal
`Adenocarcinoma #03:
`
`T IC_ Tumor weight in treated animus divided by the
`tumor weight of the control animals, reponed as per- IS
`cent value. Lower value indicates better efficacy, 0%
`sugges!.'l cures. < 10% is considered highly active, 10 to
`42% is moderately active formulation. > 42% is consid(cid:173)
`ered inactive.
`Log KilI =(T -C)/3.32 (Td), where T is the time in 20
`days for the median tumor to reach 1000 mg mass in
`treated animals, C is the time in days for the median
`tumor to reach 1000 mg in control animals and Td is the
`tumor volume doubling time in days. Cures (tumor free 25
`animals) are excluded from (T -C) calculations.
`&ample I demonstrates that a composition of this
`invention exhibited reduced toxicity and enhanced effi(cid:173)
`cacy compared to a prior art composition and could be
`admlnistered by IV bolus injection.
`
`30
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`2. Efficacy Stud