(12)
`
`United States Patent
`Bosch et al.
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`US 6,656,504 B1
`Dec. 2, 2003
`
`US006656504B1
`
`(54) NANOPARTICULATE COMPOSITIONS
`COMPRISING AMORPHOUS
`CYCLOSPORINE AND METHODS OF
`MAKING AND USING SUCH
`COMPOSITIONS
`
`(75) Inventors: H. William Bosch, Bryn MaWr, PA
`(US); Kevin D. Ostrander, Reading,
`_
`PA (Uslpouglas C- H°VeY>
`collegevllle, PA (Us)
`
`
`
`ASSIgIlee: Elan Phal‘ma IIItEI‘IIatiOIIal Ltd-, Shannon (IE)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U_S_C_ 154(k)) by 0 days_
`
`(21) App1.No.: 09/392,557
`_
`(22) F1169:
`Sep- 9’ 1999
`
`(51) Int. Cl.7 .......................... .. A61K 9/14; A61K 9/50;
`A61K 38/00
`
`(52) US. Cl. ..................... .. 424/489; 424/501; 424/502;
`514/11
`(58) Field of Search ............................... .. 424/489, 501,
`424/502; 514/11, 937
`
`(56)
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,911,099 A * 10/1975 DeFoney et 211.
`427/213.31
`4,540,602 A
`9/1985 Motoyama et a1.
`4,727,077 A
`2/1988 Haga et a1. ............... .. 514/274
`4,783,484 A 11/1988 Violante et a1.
`514/535
`4,826,689 A
`5/1989 Violanto et al.
`424/489
`4,851,421 A
`7/1989 IWasaki et al. .... ..
`514/352
`4,904,668 A
`2/1990 Kondo et a1. ...... ..
`514/274
`4,983,605 A
`1/1991 Kondo et a1. ............. .. 514/247
`4,997,454 A
`3/1991 Violante et a1. ........ .. 23/305 A
`5,002,952 A
`3/1991 Kondo et a1. ............. .. 514/274
`5,098,907 A
`3/1992 Kondo et a1. ............. .. 514/274
`5,145,684 A
`9/1992 Liversidge et al. ....... .. 424/489
`5,264,213 A 11/1993 Shibahara et a1. ........ .. 424/409
`5,389,382 A * 2/1995 List et a1.
`
`3/1995 Liversidge et a1. ....... .. 424/490
`5,399,363 A
`7/1995 June ......................... .. 424/489
`5,429,824 A
`8/1995 Desai ....................... .. 424/451
`5,439,686 A
`(List continued on next page.)
`
`FOREIGN PATENT DOCUMENTS
`
`A2
`
`g
`WO 96/14079
`W0 96/31202
`W0 97 35603
`
`21987
`/1988
`5/1996
`10/1996
`10 1997
`
`W0
`W0
`W0
`
`OTHER PUBLICATIONS
`*
`I’
`“
`_
`_
`_
`Physicians Desk Reference Neoral Sandimmune', 2000.
`Kondo et al., “Improved Oral Absorption of Enter1c Copre
`cipitates of a Poorly Soluble Drug,” J. Pharm. Sciences,
`8399566570 (1994)
`Kondo et al., “Improved Oral Absorption of a Poorly
`Water—Soluble Drug, HO—221, by Wet—Bead Milling Pro
`ducing Particles in Submicron Region,” Chem. Pharm.
`BulL, 41(4):737—740 (1993).
`Kondo, et al., “Pharmacokinetics of a MicroniZed, Poorly
`Water Soluble Drug, HO—221, in Experimental Animals,
`”Biol_ Pharm Bull” 16(8);796_8()() (1993)_
`The Merck Index: An Encyclopedia of Chemicals, Drugs,
`and Biologicals (12* Ed, 1996) pp, 464—465,
`
`Primary Examiner—Thurman K. Page
`Assistant Examiner—Amy E Pulliam
`(74) Attorney, Agent, or Firm—Foley & Lardner
`(57)
`ABSTRACT
`Nanoparticulate amorphous cyclosporine formulations, and
`nanoparticulate cyclosporine formulations comprising a
`mixture of amorphous and crystalline cyclosporine, having
`effective average particle siZes of less than about 2000 nm
`are described. The compositions exhibit increased bioavail
`ability and increased consistency of bioavailability as com
`pared to prior macro-sized cyclosporine and formulations.
`Methods of making and using the compositions are also
`described.
`
`59 Claims, 3 Drawing Sheets
`
`REPRESENTATIVE X-RAY POWDER DIFFRACTION
`PATTERN OF CRYSTALLlNE CYCLOSPORINE DRUG SUBSTANCE
`
`5000
`
`4500
`
`4000
`
`3500
`
`3000
`[eoumsl
`-
`2500
`
`2000
`
`1500
`
`1000
`
`500—
`
`40
`
`

`

`US 6,656,504 B1
`Page 2
`
`US. PATENT DOCUMENTS
`
`Liversidge et a1. ....... .. 424/490
`
`.. 424/489
`241/5
`. 424/9.411
`424/9.1
`424/493
`CZekai et a1. ............... .. 241/21
`
`5,741,522 A
`5,766,629 A
`5,776,496 A
`5,827,822 A
`5,834,017 A
`5,862,999 A
`6,228,399 B1
`
`* cited by examiner
`
`Violante et a1. .......... .. 424/489
`Cho et a1. ................. .. 424/455
`
`Violante et a1. .......... .. 424/489
`Floc’H et a1.
`.... .. 514/11
`
`424/455
`Cho et a1. .... ..
`CZekai et a1. ............... .. 241/21
`Parikh et a1.
`
`

`

`U.S. Patent
`
`Dec. 2, 2003
`
`Sheet 1 of3
`
`US 6,656,504 B1
`
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`

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`U.S. Patent
`
`Dec. 2, 2003
`
`Sheet 2 of3
`
`US 6,656,504 B1
`
`_ _ _ _ _ _
`
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`U.S. Patent
`
`Dec. 2, 2003
`
`Sheet 3 of3
`
`US 6,656,504 B1
`
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`

`US 6,656,504 B1
`
`1
`NANOPARTICULATE COMPOSITIONS
`COMPRISING AMORPHOUS
`CYCLOSPORINE AND METHODS OF
`MAKING AND USING SUCH
`COMPOSITIONS
`
`FIELD OF THE INVENTION
`
`The present invention is directed to nanoparticulate com
`positions comprising amorphous cyclosporine, or a mixture
`of amorphous and crystalline cyclosporine, and methods of
`making and using such compositions.
`
`10
`
`15
`
`BACKGROUND OF THE INVENTION
`Cyclosporine is a hydrophobic, cyclic, undecapeptide that
`exerts immunosuppressive, antiin?ammatory, antifungal,
`and antiparasitic activities. Immunosuppressive medications
`play a large part of the management of many pediatric
`illnesses. Cyclosporine is the primary tool used to prevent
`rejection folloWing solid organ and bone marroW transplan
`tation; the drug helped revolutioniZe transplantation by
`improving transplant survival, reducing hospitalization, and
`reducing patient morbidity. It has been estimated that
`cyclosporine is given to more than 90% of children Who
`have received a kidney transplant in the United States.
`Cyclosporine also has been effective in various other
`autoimmune conditions such as uveitis, psoriasis, type I
`diabetes mellitus, rheumatoid arthritis, in?ammatory boWel
`disease, certain nephropathies, refractory Crohn’s disease,
`ulcerative colitis, biliary cirrhosis, aplastic anemia, rheuma
`toid arthritis, myasthenia gravis, and dermatomyositis.
`Cyclosporine is in clinical use WorldWide under the trade
`names SANDIMMUNE® (Novartis), NEORAL®
`(Novartis), and SANGCYA® (SangStat).
`SANDIMMUNE®, introduced in 1983, suffered from poor
`and Widely variable absorption rates. This prompted devel
`opment of a second generation cyclosporine formulation,
`NEORAL®, Which is a microemulsion formulation having
`better absorption than SANDIMUNE®-both in quantity and
`consistency. Since 1995, When NEORAL® Was introduced,
`about 70% of patients have sWitched from SANDIM
`MUNE® to NEORAL®, indicating the severity of poor and
`inconsistent absorption of cyclosporine. SAN GCYA®,
`Which is a modi?ed oral solution bioequivalent to
`NEORAL®, Was introduced in 1998.
`Cyclosporine is administered orally and intravenously
`(IV). After oral administration, roughly 20 to 50% is
`absorbed, although absorption is highly variable. First-pass
`metabolism, mode of administration, and drug interactions
`all affect cyclosporine absorption. Food decreases the
`absorption of NEORAL® and SAN GCYA®.
`Cyclosporine is extremely hydrophobic. The IV formu
`lation contains 33% alcohol and a castor oil vehicle to
`solubiliZe the drug, Which is believed to account for occa
`sionally severe hypersensitivity reactions. Oral preparations
`can contain corn, castor or olive oil and ethanol, but in loWer
`concentrations. The dose normaliZed area under the curve
`(AUC) is 23% greater for NEORAL® or SANGCYA® as
`compared to SANDIMUNE® in renal transplant, rheuma
`toid arthritis, and psoriasis patients, and 50% greater in liver
`transplant patients. Data for cardiac transplant patients is
`limited, but similar increases have been noted. Increases in
`peak blood cyclosporine concentrations (NEORAL® and
`SAN GCYA® related to SANDIMMUNE®) range from 40
`to 106% for renal transplant patients and 90% for liver
`transplant patients.
`
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`While NEORAL® and SAN GCYA® are an improvement
`over SANDIMMUNE®, the conventional cyclosporine for
`mulations suffer from poor bioavailability because, among
`other things, cyclosporine is poorly Water soluble.
`Moreover, currently marketed cyclosporine formulations are
`knoWn to have disadvantageous “intersubject variability,”
`i.e., it has been found that, given the same dosage amount,
`actual blood levels of cyclosporine vary signi?cantly from
`patient to patient. See Physicians’ Desk Reference (1998) at
`1882 et seq. This represents an important shortcoming of
`these drugs. Speci?cally, because cyclosporine has a narroW
`therapeutic index (a narroW range betWeen an effective
`dosage and a harmful dosage), the inability to predict drug
`absorption requires that physicians closely monitor each
`patient to establish baseline absorption levels. Such moni
`toring is expensive and time consuming. In addition, the
`poor absorption and patient variability of knoWn cyclospo
`rine formulations can make dosage formulation dif?cult.
`Proper dosage formulation for cyclosporine is critical
`because the drug is a general immunosuppressive.
`Therefore, the drug can result in an increased susceptibility
`to infection. Too much drug can result in uncontrolled
`infection While too little can result in organ rejection.
`One drug delivery method that can result in increasing the
`bioavailability, increasing the absorption rate, quantity, and
`consistency, and decreasing the toxicity of a drug is formu
`lation of the drug into a nanoparticulate composition. Nano
`particulate compositions, ?rst described in US. Pat. No.
`5,145,684 (“the ’684 patent”), are particles consisting of a
`poorly soluble crystalline therapeutic or diagnostic agent
`onto Which are adsorbed a non-crosslinked surface stabi
`liZer. Nanoparticulate compositions comprising cyclospo
`rine are not described by the ’684 patent. Nanoparticulate
`compositions containing crystalline, but not amorphous,
`cyclosporine are disclosed in US. Pat. Nos. 5,494,683 and
`5,399,363.
`Conventional large particle siZed amorphous cyclosporine
`compositions are described in US. Pat. Nos. 5,389,382 (“the
`’382 patent”) and 5,827,822 (“the ’822 patent”). These
`disclosures suffer from various de?ciencies. For example,
`the ’382 patent describes hydrosols of cyclosporine in an
`intravenous applicable, stabiliZed, pharmaceutically accept
`able form, Which is suspended or dry. The hydrosol formu
`lations are obtained by controlled precipitation methods.
`Such methods are disadvantageous in that they result in solid
`dose formulations having a loW drug to surface stabiliZer
`ratio, and liquid dispersion formulations having a loW solid
`content. This is because controlled precipitation methods
`require an excess amount of surface stabiliZer and Water to
`produce small-siZed precipitated particles. The excess of
`surface stabiliZer produces solid dose compositions having a
`large quantity of surface stabiliZer and a small quantity of
`drug, and the excess of Water produces a liquid dispersion
`formulation having a loW solids content and, therefore, a loW
`drug content.
`A high drug content for a solid dose or liquid dispersion
`formulation is preferred because it produces a more con
`centrated dosage formulation. Concentrated dosage forms of
`cyclosporine are particularly desirable because the dosage
`for this drug is relatively high, i.e., about 100 mg a day or
`more. A dosage formulation having a loW drug content, but
`requiring a high daily dosage, results in either a large pill,
`capsule, or quantity of ?uid, or multiple doses of such
`formulations, to be administered to the patient. In contrast,
`a concentrated dosage form alloWs minimiZation of the siZe
`of the orally administered pill or capsule or number of daily
`administrations.
`
`

`

`US 6,656,504 B1
`
`3
`The ’822 patent is directed to aqueous suspension formu
`lations of amorphous cyclosporin A containing lower
`alkanols as solubiliZing agents and a polyoxyalkylene sur
`factant. The addition of alcohol solubiliZing agents is fre
`quently undesirable because they can an trigger an allergic
`response in a patient. Such solubiliZing agents are often
`required for prior art cyclosporine compositions to increase
`the solubility of the cyclosporine. A drug must be absorbed
`by a patient prior to taking effect. Thus, often pharmaceu
`tical formulations of highly insoluble drugs additionally
`contain solubiliZing agents to aid in absorption of the drug
`folloWing administration.
`There remains a need in the art for cyclosporine formu
`lations that can be delivered in high dosage formulations,
`that exhibit consistent and effective absorption, that have
`decreased toxicity as compared to knoWn cyclosporine
`formulations, and Which do not require the presence of
`alcohol solubiliZing agents. The present invention satis?es
`these needs.
`
`SUMMARY OF THE INVENTION
`
`The present invention is directed to nanoparticulate com
`positions of amorphous cyclosporine and, adsorbed to the
`surface of the cyclosporine, at least one non-crosslinked
`surface stabiliZer. The cyclosporine particles of the nano
`particulate composition have an effective average particle
`siZe of less than about 2000 nm.
`In another embodiment, the invention encompasses nano
`particulate compositions of a mixture of amorphous and
`crystalline cyclosporine and, adsorbed to the surface of the
`cyclosporine, at least one non-crosslinked surface stabiliZer.
`The cyclosporine particles of the nanoparticulate composi
`tion have an effective average particle siZe of less than about
`2000 nm.
`Another aspect of the invention is directed to pharmaceu
`tical compositions comprising one or more nanoparticulate
`compositions of the invention. The pharmaceutical compo
`sition preferably comprises a nanoparticulate composition
`described above and a pharmaceutically acceptable carrier,
`as Well as any desired excipients. The compositions, Which
`can be delivered in high dosage formulations, provide for
`improved consistency of cyclosporine absorption from
`patient to patient for a given dosage amount, exhibit
`decreased toxicity, and exhibit increased absorption as com
`pared to conventional cyclosporine formulations.
`This invention further discloses methods of making nano
`particulate compositions according to the invention. A ?rst
`method comprises contacting amorphous cyclosporine, or a
`mixture of amorphous and crystalline cyclosporine, With at
`least one surface stabiliZer for a time and under conditions
`sufficient to provide a stable nanoparticulate composition.
`The surface stabiliZer can be contacted With the cyclosporine
`particles either before, during, or after siZe reduction of the
`cyclosporine particles. The cyclosporine particles of the
`nanoparticulate composition have an effective average par
`ticle siZe of less than about 2000 nm.
`The present invention is further directed to methods of
`treatment comprising administering to a mammal in need a
`therapeutically effective amount of a nanoparticulate com
`position according to the invention. The nanoparticulate
`cyclosporine composition can be administered via any con
`ventional route.
`Both the foregoing general description and the folloWing
`detailed description are exemplary and explanatory and are
`intended to provide further explanation of the invention as
`claimed. Other objects, advantages, and novel features Will
`
`10
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`4
`be readily apparent to those skilled in the art from the
`folloWing detailed description of the invention.
`
`BRIEF DESCRIPTION OF THE FIGURES
`FIG. 1: ShoWs the results of x-ray poWder diffraction of
`raW cyclosporine drug substance;
`FIG. 2: ShoWs the results of x-ray poWder diffraction of
`a milled cyclosporine formulation having Pluronic® F108 as
`a surface stabiliZer; and
`FIG. 3: ShoWs the results of x-ray poWder diffraction of
`a milled cyclosporine formulation having HPC-SL as a
`surface stabiliZer.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`The present invention is directed to compositions com
`prising nanoparticulate amorphous cyclosporine, or a mix
`ture of amorphous and crystalline cyclosporine, and meth
`ods of making and using such nanoparticulate compositions.
`As used herein, singular terms are used for simplicity of
`expression only and are not intended to limit the invention
`or aspects of the invention to singular embodiments. Thus,
`the description of, for example, “a surface stabilizer” is
`meant to describe “one or more” surface stabiliZers unless
`explicitly indicated otherWise.
`Prior to the present invention, it Was knoWn that crystal
`line drugs could be formulated into nanoparticulate
`compositions, as taught by the ’684 patent. In such
`compositions, a surface stabiliZer adsorbs to the crystalline
`surface of the drug and acts as a steric barrier to other drug
`particles to prevent agglomeration. This results in a stable
`nanoparticulate composition, in Which the particle siZe of
`the composition does not signi?cantly increase over time via
`solubiliZation and recrystalliZation or agglomeration.
`Because the surface stabiliZer adsorbs to the surface of the
`crystalline drug, and does not chemically interact With the
`drug, it Was thought that amorphous drugs could not be
`utiliZed in nanoparticulate compositions described by the
`’684 patent. Amorphous drugs do not have an intermolecular
`lattice structure, Which is characteristic of the crystalline
`solid state. Surprisingly, it Was discovered that amorphous
`cyclosporine can be incorporated into a nanoparticulate
`composition.
`An amorphous compound has a higher energy level than
`a crystalline compound. Because of this, an amorphous
`compound is generally unstable, as in nature the compound
`prefers to convert to the loWer energy crystalline state.
`Because amorphous compounds have a higher energy level
`than crystalline compounds, it is preferable that a drug be in
`an amorphous state. The amorphous state is less stable than
`the crystalline state; therefore, a solid Will be more soluble
`in the amorphous state than in the crystalline state. Improved
`solubility Will lead to rapid and more complete dissolution,
`and in the case of a poorly soluble drug substance, improved
`bioavailability.
`A. Compositions
`The compositions of the invention comprise nanoparticu
`late amorphous cyclosporine, or a mixture of amorphous and
`crystalline cyclosporine, having one or more surface stabi
`liZers adsorbed to the surface of the cyclosporine. Surface
`stabiliZers useful herein physically adhere to the surface of
`the nanoparticulate cyclosporine but do not chemically react
`With the cyclosporine itself. Individually adsorbed mol
`ecules of the surface stabiliZer are essentially free of inter
`molecular crosslinkages.
`The present invention also includes nanoparticulate com
`positions formulated into compositions together With one or
`
`

`

`US 6,656,504 B1
`
`5
`more non-toxic physiologically acceptable carriers,
`adjuvants, or vehicles, collectively referred to as carriers, for
`parenteral injection, for oral administration, for rectal or
`topical administration, or the like. The present invention
`further includes nanoparticulate compositions in solid dose
`formulations and liquid dispersion formulations.
`1. Cyclosporine
`The cyclosporins comprise a class of cyclic non-polar
`oligopeptides having valuable immunosuppressive, anti
`in?ammatory, and anti-parasitic activity. The ?rst of the
`cyclosporins to be isolated and What has been termed the
`“parent” compound of the class is the naturally occurring
`fungal metabolite referred to simply as “cyclosporine” or as
`“cyclosporin A.”
`Since the discovery of cyclosporin A, a Wide variety of
`naturally occurring cyclosporins have been isolated and
`identi?ed and other non-naturally occurring cyclosporins
`have been prepared by synthetic means or via modi?ed
`culture techniques. Such compounds are knoWn in the art
`and are described, for example, in US. Pat. No. 5,389,382
`and in The Mercklndex (12th ed. 1996) at 464—465. As used
`herein, the term cyclosporine is meant to include both
`cyclosporin A and other cyclosporins, such as cyclosporins
`B through I and synthetic analogues thereof. The preferred
`cyclosporin used herein is cyclosporin A.
`The cyclosporine compositions of the present invention
`are either partially or predominantly amorphous in nature.
`This is so even though the starting cyclosporine compound
`used to obtain the nanoparticulate compositions may be
`predominantly crystalline in nature. The term “amorphous”
`is a term With a recogniZed meaning in the chemical arts and
`describes a structure that is non-crystalline, i.e., a structure
`that lacks an intermolecular lattice structure. Whether the
`nanoparticulate composition is in a crystalline or amorphous
`state can be determined, for example, by X-ray poWder
`diffraction patterns or other methods knoWn to the skilled
`artisan.
`2. Surface StabiliZers
`Suitable surface stabiliZers can preferably be selected
`from knoWn organic and inorganic pharmaceutical excipi
`ents. Such excipients include various polymers, loW molecu
`lar Weight oligomers, natural products, and surfactants.
`Preferred surface stabiliZers include nonionic and ionic
`surfactants. TWo or more surface stabiliZers can be used in
`combination.
`Representative examples of surface stabiliZers include
`cetyl pyridinium chloride, gelatin, casein, lecithin
`(phosphatides), dextran, glycerol, gum acacia, cholesterol,
`tragacanth, stearic acid, benZalkonium chloride, calcium
`stearate, glycerol monostearate, cetostearyl alcohol,
`cetomacrogol emulsifying Wax, sorbitan esters, polyoxyeth
`ylene alkyl ethers (e.g., macrogol ethers such as cetomac
`rogol 1000), polyoxyethylene castor oil derivatives, poly
`oxyethylene sorbitan fatty acid esters (e.g., the
`commercially available TWeens® such as e.g., TWeen 20®
`and TWeen 80® (ICI Specialty Chemicals)); polyethylene
`glycols (e.g., CarboWaxs 3350® and 1450®, and Carbopol
`934® (Union Carbide)), dodecyl trimethyl ammonium
`bromide, polyoxyethylene stearates, colloidal silicon
`dioxide, phosphates, sodium dodecylsulfate, carboxymeth
`ylcellulose calcium, hydroxypropyl celluloses (e.g., HPC,
`HPC-SL, and HPC-L), hydroxypropyl methylcellulose
`(HPMC), carboxymethylcellulose sodium, methylcellulose,
`hydroxyethylcellulose, hydroxypropylcellulose,
`hydroxypropylmethyl-cellulose phthalate, noncrystalline
`cellulose, magnesium aluminum silicate, triethanolamine,
`polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), 4-(1,
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`1,3,3-tetramethylbutyl)-phenol polymer With ethylene oxide
`and formaldehyde (also knoWn as tyloxapol), poloxamers
`(e.g., Pluronics F68® and F108®, Which are block copoly
`mers of ethylene oxide and propylene oxide); poloxamines
`(e.g., Tetronic 908®, also knoWn as Poloxamine 908®,
`Which is a tetrafunctional block copolymer derived from
`sequential addition of propylene oxide and ethylene oxide to
`ethylenediamine (BASF Corporation, Parsippany, N] a
`charged phospholipid such as dimyristoyl phophatidyl
`glycerol, dioctylsulfosuccinate (DOSS); Tetronic 1508®
`(T-1508) (BASF Corporation), dialkylesters of sodium sul
`fosuccinic acid (e.g., Aerosol OT®, Which is a dioctyl ester
`of sodium sulfosuccinic acid (American Cyanamid));
`Duponol P®, Which is a sodium lauryl sulfate (DuPont);
`Tritons X-200®, Which is an alkyl aryl polyether sulfonate
`(Rohm and Haas); Crodestas F-110®, Which is a mixture of
`sucrose stearate and sucrose distearate (Croda Inc.);
`p-isononylphenoxypoly-(glycidol), also knoWn as Olin
`lOG® or Surfactant 10-G® (Olin Chemicals, Stamford,
`Conn.); Crodestas SL-40® (Croda, Inc.); and SA9OHCO,
`Which is C18H37CH2(CON(CH3)—CH2(CHOH)4(CH2OH)2
`(Eastman Kodak Co.), triblock copolymers of the structure
`-(-PEO)--(-PBO-)--(-PEO-)- (knoWn as B20—5000), and the
`like.
`Most of these surface stabiliZers are knoWn pharmaceu
`tical excipients and are described in detail in the Handbook
`of Pharmaceutical Excipients, published jointly by the
`American Pharmaceutical Association and The Pharmaceu
`tical Society of Great Britain (The Pharmaceutical Press,
`1986), speci?cally incorporated by reference. The surface
`stabiliZers are commercially available and/or can be pre
`pared by techniques knoWn in the art.
`The invention includes that each of the above-described
`stabiliZers or other stabiliZers described herein or described
`in a reference cited herein can be used either alone, in
`combination With each other, or With other surface stabiliZ
`ers.
`3. Nanoparticulate Particle SiZe
`Preferably, the compositions of the invention contain
`nanoparticles Which have an effective average particle siZe
`of less than about 2000 nm, less than about 1500 nm, less
`than about 1000 nm, less than about 800 nm, less than about
`700 nm, less than about 600 nm, less than about 500 nm, less
`than about 400 nm, less than about 300 nm, less than about
`200 nm, less than about 100 nm, or less than about 50 nm,
`as measured by light-scattering methods or other methods
`accepted in the art. By “an effective average particle siZe of
`less than about 2000 nm” it is meant that at least 50% of the
`drug particles have a Weight average particle siZe of less than
`about 2000 nm When measured by light scattering or other
`conventional techniques. Preferably, at least 70% of the drug
`particles have an average particle siZe of less than about
`2000 nm, more preferably at least 90% of the drug particles
`have an average particle siZe of less than about 2000 nm, and
`even more preferably at least about 95% of the particles have
`a Weight average particle siZe of less than about 2000 nm.
`4. Concentration of Cyclosporine and Surface StabiliZer
`Preferable ratios of cyclosporine to surface stabiliZer are
`about 10:1 to about 1.511, by Weight. With liquid
`dispersions, preferred drug content is about 50% to about
`2% by Weight.
`B. Methods of Making Nanoparticulate Formulations
`Exemplary methods of making nanoparticulate composi
`tions are described in the ’684 patent. The optimal effective
`average particle siZe of the invention can be obtained by
`controlling the process of particle siZe reduction, such as by
`controlling the milling time and the amount of surface
`
`

`

`US 6,656,504 B1
`
`7
`stabilizer added. Particle growth and particle aggregation
`can also be minimized by milling the composition under
`colder temperatures and by storing the ?nal composition at
`colder temperatures.
`Milling to obtain a nanoparticulate composition com
`prises dispersing cyclosporine particles in a liquid dispersion
`medium, folloWed by applying mechanical means in the
`presence of grinding media to reduce the particle siZe of the
`cyclosporine to the desired effective average particle siZe.
`The cyclosporine particles can be reduced in siZe in the
`presence of one or more surface stabiliZers. Alternatively,
`the cyclosporine particles can be contacted With one or more
`surface stabiliZers after attrition. Other compounds, such as
`a diluent, can be added to the cyclosporine/surface stabiliZer
`composition during the siZe reduction process. Dispersions
`can be manufactured continuously or in a batch mode. The
`resultant nanoparticulate cyclosporine dispersion can be
`utiliZed in solid or liquid dosage formulations. Exemplary
`useful mills include loW energy mills, such as a roller or ball
`mill, and high energy mills, such as Dyno mills, NetZsch
`mills, DC mills, and Planetary mills.
`The starting cyclosporine composition can be predomi
`nantly crystalline, predominantly amorphous, or a mixture
`thereof. The resultant cyclosporine composition is predomi
`nantly amorphous.
`Asolid dosage formulation can be prepared by drying the
`nanoparticulate amorphous cyclosporine, or mixture of
`amorphous and crystalline cyclosporine, folloWing grinding.
`Apreferred drying method is spray drying. The spray drying
`process is used to obtain a nanoparticulate poWder folloWing
`the milling process used to transform the cyclosporine into
`nanoparticles. Such a nanoparticulate poWder can be formu
`lated into tablets for oral administration.
`C. Methods of Using Nanoparticulate Compositions of the
`Invention
`The nanoparticulate compositions of the invention can be
`administered to humans and animals either orally, rectally,
`parenterally (intravenous, intramuscular, or subcutaneous),
`intracistemally, intravaginally, intraperitoneally, locally
`(poWders, ointments or drops), or as a buccal or nasal spray.
`Compositions suitable for parenteral injection may com
`prise physiologically acceptable sterile aqueous or nonaque
`ous solutions, dispersions, suspensions or emulsions and
`sterile poWders for reconstitution into sterile injectable solu
`tions or dispersions. Examples of suitable aqueous and
`nonaqueous carriers, diluents, solvents, or vehicles includ
`ing Water, ethanol, polyols (propyleneglycol,
`polyethyleneglycol, glycerol, and the like), suitable mixtures
`thereof, vegetable oils (such as olive oil) and injectable
`organic esters such as ethyl oleate. Proper ?uidity can be
`maintained, for example, by the use of a coating such as
`lecithin, by the maintenance of the required particle siZe in
`the case of dispersions and by the use of surfactants.
`The nanoparticulate compositions may also contain adju
`vants such as preserving, Wetting, emulsifying, and dispens
`ing agents. Prevention of the groWth of microorganisms can
`be ensured by various antibacterial and antifungal agents,
`such as parabens, chlorobutanol, phenol, sorbic acid, and the
`like. It may also be desirable to include isotonic agents, such
`as sugars, sodium chloride, and the like. Prolonged absorp
`tion of the injectable pharmaceutical form can be brought
`about by the use of agents delaying absorption, such as
`aluminum monostearate and gelatin.
`Solid dosage forms for oral administration include
`capsules, tablets, pills, poWders, and granules. In such solid
`dosage forms, the active compound is admixed With at least
`one of the folloWing: (a) one or more inert excipients (or
`
`8
`carrier), such as dicalcium phosphate; (b) ?llers or
`extenders, such as starches, lactose, sucrose, glucose,
`mannitol, and silicic acid; (c) binders, such as
`carboxymethylcellulose, alignates, gelatin,
`polyvinylpyrrolidone, sucrose and acacia; (d) humectants,
`such as glycerol; (e) disintegrating agents, such as agar-agar,
`calcium carbonate, potato or tapioca starch, alginic acid,
`certain complex silicates, and sodium carbonate;
`solution
`retarders, such as paraf?n; (g) absorption accelerators, such
`as quaternary ammonium compounds; (h) Wetting agents,
`
`such as cetyl alcohol and glycerol monostearate; adsorbents, such as kaolin and bentonite; and lubricants,
`
`
`such as talc, calcium stearate, magnesium stearate, solid
`polyethylene glycols, sodium lauryl sulfate, or mixtures
`thereof. For capsules, tablets, and pills, the dosage forms
`may also comprise buffering agents.
`Liquid dosage forms for oral administration include phar
`maceutically acceptable emulsions, solutions, suspensions,
`syrups, and elixirs. In addition to the active compounds, the
`liquid dosage forms may comprise inert diluents commonly
`used in the art, such as Water or other solvents, solubiliZing
`agents, and emulsi?ers. Exemplary emulsi?ers are ethyl
`alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate,
`benZyl alcohol, benZyl benZoate, propyleneglycol, 1,3
`butyleneglycol, dimethylformamide, oils, such as cotton
`seed oil, groundnut oil, corn germ oil, olive oil, castor oil,
`and sesame oil, glycerol, tetrahydrofurfuryl alcohol,
`polyethyleneglycols, fatty acid esters of sorbitan, or mix
`tures of these substances, and the like.
`Besides such inert diluents, the composition can also
`include adjuvants, such as Wetting agents, emulsifying and
`suspending agents, sWeetening, ?avoring, and perfuming
`agents.
`Actual dosage levels of active ingredients in the nano
`particulate compositions of the invention may be varied to
`obtain an amount of active ingredient that is effective to
`obtain a desired therapeutic response for a particular com
`position and method of administration. The selected dosage
`level therefore depends upon the desired therapeutic effect,
`on the route of administration, on the desired duration of
`treatment, and other factors.
`The total daily dose of the compounds of this invention
`administered to a host in single or divided dose can vary
`Widely depending upon a variety of factors, including the
`body Weight, general health, sex, diet, time and route of
`administration, rates of absorption and excretion, combina
`tion With other drugs, and the severity of the particular
`condition being treated