AU2006200768B2 - Phosopholipid-based powders for drug delivery - Google Patents

Description

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT Applicant: ALLIANCE PHARMACEUTICAL CORP.

Invention Title: PHOSOPHOLIPID-BASED POWDERS FOR DRUG DELIVERY The following statement is a full description of this invention, including the best method of performing it known to me/us: PHOSPHOLPID-BASED POWDERS FOR DRUGy DEIVERY Field of t hIvention The present invention relates to, particua-t compositions suitable for drug delivory, prefenbly via inbalsrion. in particular, the present invention provides phospholipidconmbang Particulate coniPositions cowprsing a polyvalant cation. The particulaie compositions of the present invention exhibi an ineased gel-to-liquid crystal tmnsition tempera zresulting in =mproved disperdNb!it and storage stbiity.

Euckpooud of the Invention Phospholipids are major component of cell and orgmnelle membranes, blood ipOoteis, and lung surfacwa. in fter= of pulnonary ring delivery, ph oplds have been investigate as therapeuic W=nt for the treatmen of respirtor distres syndrom (La- exogenus lung sUlfactants), and as suitable excipiwnis for the delivery of actives. The intracton. of phaspholipids with walr is critcal to thef formalon, maintenance, and function of each Of these iportan biooicl comlexe (Mcntos and Magid). At low t= in lbm ge phase, the acyl chains are in a a fmtinly welktodared stai, esasnaf aly n the all-han Configuration. At highe temperatures, above t chatin znlth tzmpeaxr, ts Chain order is lost, owing to an imase in gauche confomuer cct (Seddon and Cove).

Seve exogenous lung amtctauu have bee maketed and iclude prodct derived from bovimplug (Suryn I. Abbott Laboratorie), porcilup (Cmo-amfl, Dey LabOnzoRtie4) or compltly synthetic urfactantg with no apoproteiw

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EXoSurM QOlo Welicome). To data. Ihese prodnats have been utiliwi for te ieulnut of infan respiatory distfre Yadromne (IRD). Now have bean successfu in reciving FDA w'EM"I for the ftren of adult respamory distres syndrome (AIDS). Thecen ifmdose is 100 mg&- Fam a S0 kg adult this would tranlat int £m daiof 5g. A, doam a3f thin mount can only be administered to ARIDS paient by diMAc haulzl into the Va patimt's endoiachtal tube, or possibly via nabulization of aqueous dispersions of the CI 5sfctant mmdwiil, Instilation of sfnatans leads to deposition primarily in the =enal airways, and liztle of the drug makes it to the alveoli, where it is needed toimprove gas exchange in these caitically ill patients. Nebulizaton of smfactant may allow for greater peripheral delivery, but is plagued by the fact that curenr nebulizers are inefficient devices and 00 only ca- 10% of the drug actually reaches the patients luagin thfe suractant solutions IN foam during the nebulization process, leading to conplications and frther loss of drag. It o is believed tat as mch as 99% of the admkninme smfactnt may be wasted due to poor delivery to the patient If mor effective delivery of snractart could be achieved, it likely Va Ihat the adrinistered dose and cost for trvanent of ARDS could be dramatically decrtased.

Fther, lung stufactant has been shown to modulate mucous cmnsppo in airways.

In this regard, the chronic administrarion of sfactant fdr the treatment of patients with chronic obstructive pulmonary disease (COPD) has been suggested. St other iicatiows with significandy lower doses may be open to tzeazmeif a dy powdr form of a lung smtztant were available. The powdered srzctat fonmulstion may be purely synhed with no added apopromins). Alteratively, the powder fonnulation could contain the hydrophobic apoproteins SP-B or SP-C or alternative recombinant or synthzic peptide ndmetics (eg. 0).

Due to its spreading charareristlcs on lung epithelia. srfamant has been proposed as the ideal carer for delivery of drugs to the lg and via the lung to the systemic circulation. Once again, achieving efficient delivery to the lung is important, ospecially in LiHt of the potential high cost of many of the cmunt products. One potential way to deliver drugs in phospholipids is as a dry powder aerosolirad to the lung. Mot fe powders 5np) eahibit poor dipemibfity. This ca be problematic when anwnpting to delivezM awsoize, and/or package the powders.

The major frcs that control particle-particle inteactions can be divided into short and long =gs forces. Lnngrag fores inclde gravational aftive foes and eleciustadcs, whm the itercton vaies as the square of te searation ditac Shortruge acactive fames dominate for dry powders and include vun der Whals intucdovs, hydrogen bonding. and iquid brdging. Liquid bridging oew when wate nwlecules axe able to iheveruibly hind paticles together.

Phospholipids ar especially difficult to formIlate as dry powders as thefr low gel to hquid cyswtal tranidan tenpermnn m) vales and amorphou nafl lead to powders which am very sticky and diMcuk to deaggregate and aemsoli Phospholipids with Tm

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o values less than 10C egg PC or any usatnated lipids) form highly cohesive powders following spray-drying. Inspection of the powders via scanning electron microscopy reveals highly agglomeaated particles with surfaces that appear to have been melted/annealed. Fornnnlaing phosphoipid powders which have low Tm are problematic, especially if one hopes to achieve a certain particle morphology, as in the case of aerosol delivery. Thus, it would be advantageous to find ways to elevate the Tm of these lipids.

00 Examples of particulate compositions incorporating a surfacuar are disclosed in PCr publications WO 99/16419, WO 99/38493, WO 99/66903, WO 0Q10541, and U.S. Patent 0 Nos. 5,855,913. which are bereby incorporated in their enthiety by refaen.

Currently, lung surfactant is given to patients by intabating them and instilling a Suspension of lUng surfactant directly into the lungs. This is a highly invasive procedure which generally is not performed on conscious patients. and as do most procedures, cardies its own risM. Potential applications for lung surfactant beyond the crment indication of respiratory distress syndrome in neonates are greatly limited by this method of administration. For example, lung surfactam may be useful in a variety of disease states that are, in part, due to decreased nlung saurfactant being present in the lungs. U.S. Patnt Nos. 5,451,569, 5,698,537, and 5,925,337. and PCT publications WO 97/26863 and WO 0027360. for example, disclose the pulmanary administration of luntg murfactant to beat various conditions, the disclosures of which are hereby incorporated in their entirety by reference. Diseases that are thought to be possibly aggravated by lung surfactmt deficiency include cystic fibrosis, chronic obsnuctive pulmonary disease, and asthma, just to name a few. lbs delivery of exogenous hUmg surfactant, in a topical fashion, to patients suffering from these diseases may amelirate certain signs and symptoms of the diseases.

For chronic conditions, the regular (once or more times per day on a prolonged basis) delivery of lunhmg surfactant via intabation and instillation to ambulatory patiaents is impractical Furthr, because oftheir high surface activity, lag smurfactat suspensions are not amenable to nebalimtion due to fdbaming. The current delivery of phospholipid based preparation by instillation or nebnlization are highly ineffient in delivering material to te peripheral lung. Therefor, the ability to deiver ng surfactat to patients via dry powder inhalation would be a tremendous advantage over the curent metod, since it would avoid the need for intnhation, thereby expanding the potential uses of lng sudactant i fin the ciical setting.

S Summnary of the Invention The Present invention provides far dry powder compositions of phospholiid suitabl. for drug delvery. According to a preferred embodiment the phospbolipid compositions ane efficiently delivered to the deep lung. The phosphalipid may be delivered alone, as in the case of lung suxtant or in combinaton with anothcr active age=t and/r 00 excipiwnz The use of dry powder compositions may also open new indications for use Va ~since the patient need not be intuatd. According to one embodiuxen 4 the compositions of ft prsn ineto =y be deiee from a simple passiv DPI device The preset ci 10 compositons allow for greter stabiiy on tae, and for nm efficient delivery to the 0 ~It has been fomad in the present work that the gel to liquid crystal ph=s transition of the phospholipid, Tm, is crlf eat i= obtaining phosphalipid -based dry powders that both flow well, and are readily dispemsble from a dry powder ihaer device The pr=sen invention is minated to t use of Polyvalemt cations, preferably divalent cation, to dramaticaily incrase the T)M Of phoSpholipids. AS use herein, "polyValan Cazions" refer to polyvalent Ealts or their ionic cowpoems kmsing the Tin of the phosphalipid leads to d-e folowing forinubniou impravemms: Incrase, in Tm allows the fanlator to increae the iWet and oudiet =eau on the spry-drier, or on a vacumn oven daring a secondary drying step. Hgher temperatutres allow the drying phase of the spray-drying to be controllable over at wider teupran rainge, thereby facilitating ivuval of trapped blowing agent used in the maufacture of powders accbntling toane aspect of the present invention; Incrcase in TM allow for a large differnc between Tm and the s=rage thrb mroving powder nabilitry Incrase in Tm yield phaspholipids in the gel State, wher they are less prone to raking up wate and water bridging pheuomcn Inceass in Tm yield phospholiids which arm able to spread tm efrdvely qpon COnta= with lung epitilia tha hydrated phoepholipids,, Urreby allowing drugs to be mom effectively diawiburd to the lung pedpbeiy Inanes in Tmn faInwxoves the disperlilty of the ruIting powders, tereby improving the emitted dame and fine particle f raction following pulmonary delivery.

According to a preered embodiment, the promen inventi relates to highly dispersible dry powder compositions of phospholipds suitabl for pulmnarydelivary. The compositons according to the preen invenfij =r usefu as synhetc lung mrfcta fir fit Zemwnz of local hung coditions g.Astam, COPE)) of as cari= ftthe pbulmnay delivery of acte aguats, including Small MOlCules peptdes, potein, D)NA, and itmuologi am nw o One aspecr of the Present invention is to provide powdered, dispersible compos tran having stable disperui-biliwy over time. The compositions exhbit a characteristic gel to liquid crystal phase transition remperan, Tm= which is grmaer tian a re-ommended stirage tempmeraar Ts, typically room temperaae, by at least 20 C.

Preferably Tm. is at least 40 tC greaer than Ts.

It is a further aspect of the present invenion that the increases in TM afforded by 00 addition of divalent cations leads to the abiliy to dry the powders in a seconadary drying VaO ste at temPeratre up to the Tm. of the lipid. As well, it is possible to increase t oli an oule teprbr on a spray-drier should a spray-day process be employed (TPd Va 10 ~Tm), 0 It Ws a furthe Aspect of the present invention to provide. a powdered, dispersible fc= Of a lung smxtaant having stable dispersibility over time and &eelent spreading characreristics on an aqueous subphase.

It is a further aspect of the present invention thatteipiew in dispersibility obtained by the present compositions allow for a simple, passive inhaler device to be utilized, in spite Of the fact that panictes less than 5 gm. ame conrenplated and generelly Preferred. Presenlt scare-of-thc-ar for=uad=on for fine particles Utilize blends with large lactose- Particles to imve dispersibility. When placed in a passive DPI device such fcmulaionS exhibit a sting dependence of emitted dose and hang deposition on the patiets insPiratory tlowrate. The Present compositions exibit lite. flwrat dependence cu the emitted dose and lung deposition.

Brief Descripimo of the Drawings 2S Fle I is a graph depicting the physical stablity of budesnde in pMDt- FM=ur 2 are SEM photographs the effect of calcium ion conceunurio on the £mnpbology of sny-dried particles. accrding to th eneiotL Fgue 3 is a graph depicting the sprading charaterst of powders of the instant so Definition "Active agent' as da&erbed herein includes an agent, drag, compotud, of matter Ofor lmxue thmeo wich provides some dagnostic, prophylactc, or phMmua 9cgc often benefcia, s~ct. This inchles Rfoos food suppl=Mens nuteaM drOp. vaccine, vitamins, and other beneficial agets. As used herein, the term furthe ichade 517Y PhYSz1logicahy Or pharzncoiogically active mubtbace that produce a localized Or syM Ic= effec in a. pleuL The activ aget thatcan be delvered incudes antibiotics o ilfbodiea, wtivml agens mnepfilaptics, anagesics, ant-ui.ainfmatorY agents and brauebodlnxca, and viruse and May be inorganic and organic compounds, including, O wkhou±L lini Z 1 dugs which act om the peripheral nerves, adwnergic receptors, cholinergIc receptors, the skeletal mnuscles, the cardiovascular systc, smoot amsles, thle blood circulatory systm,. synaptic sites nearoector junciona sites endocrie and horn, systems, the inmmunological system, the reproductive systm, the sken system 00autacoid systMs, the amentary and excretory sys tem, histaine system ad the ceifral nAvous system. Suitable agents my be selected from, for eamnple, olysacc2~huijdes, steroids, hypnotics, and sedaties, psychic enerizer, tunquilizem, auticonvulaans muscle relnr, adaf=agents, analgesics, and mi)de o rz~~MUSCccnffaCran, antimicrobials, antimnalarials, honionalaet including 0 ~coniraceptives, syinpathomiintics, polypeptides, and proteins capable of elicitg physiologica effects, diuretics, lipid regulating age=t, amiandrogenic agent, antpanasnncs, neoplastcs, anieoistc. hypoglycemics, nutritional agent and supplements, growth supplementsfts, antienteitis age=t, electrolyts, vaccines and diagostic agents.

EsMples of active agents useful in this invention =icude but are not limited to Minsuin calcionn. erydhropoicdin (220), Ficto VIII Factor MX ceredase, cerezyae cyclospodue graulocyte colony stiniing factor alpba-I proteinase inbibitor, elatonin, graulocyce maczuhag colony stimiating factor (GMCSp) grwth human growth hormoe (hCli), growth hormon raleasing horame (CI! heparin, low lecular Weig htpain (LMWH), intefeon Hlpha, imneeron bo1a, inerfron gamm inrlrdukiu-2, lurainizn hormone releasing hormone (TRM, leurolide, sonizosatn, smotsad analogs including octroodde, vasopxusi analog, follie stmulabin horm (FH) moglobuiius insulmiie growh factor insulflopi, inteekmIa mrecp=o antagonist, htaieki..3, iutsrlcukIn-4 iterieukhn.4, macrnpbage colony seinwilazing fria (M-CSF), namv growth factor parathroid hormone (PFm) thymsin alpha 1.

IIWHt hibzrtor. alpha-I azflrYPsi, re~paphtoy synoytWa virus antiody, cystic fibrosis 1. rgm~t- (C~FlL) gene, dWXn-tonualn (floa).

bWaccida~pevai bfn~It 1 inceaing potein OM and-CNX antibody, iniurleukin-l ueceptr 13-cas rat= acid, nicotine nicote biaze=, aip=ncrofioxai, anyhozdin, amikacin, tobnumyin penraidiun iaeduiome, albuterl sulfate, nflprucreol sulfate, bW fi m i amooaReaadbdsn aWnkuWe ipr*Uiri bromide, fiunisolide, fluti=Casone fiutiasane piopionata, mauglno xinofoWA, fonezerol fil"Axate, cnmuoyn sodium ergotaii a and the analogues agumst and wgns of t above. Activ agents; may fthrcomwise nucleic adids,

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O present as bare nucleic acid molecules, viral vectors, associated viral particles, nucleic acids associated or incorporared within lipids or a lipid-containing material. plasmid DNA )or RNA or other nucleic acid construction of a type suitable for transfetin or transformation of cells, particularly cells of the alveolar regins of the lngs. The acdve C-I 5 agents may be in various faormn, such as soluble and insoluble charged or uncharged molecules, components of molecular complexes or pharmacologically acceptable salts.

00 The active agents may be naturally occuring molecules or they may be recominantly Va produced, or they may be analogs of the natwally occunting orrecombiantly pmduced active agents with one or more amino acids added or deleted. Fther. the active agent may comprise live attenuated or killed viruses suitable for use as vaccines.

o As used herein, the tem "emitted dose" or 'ED" refers to an indication of the 0 delivery of dry powder from a suitable inhaler device after a firing or dispersion event from a powder unit orreservoir. iED is defined as the ratio.of the dose delivered by an inhaler device (described in detail below) to the nomtninal dose (Le, the mass of powder per unit dose placed into a suimtable inhaler device prior to firing) The ED is an experimentallydeterined amount, and is typically detennined using an in-vitro device set up which mimics patient dosing. To determine an ED value, a nominal dose of dry powder (as defined above) is placed into a suitable dry powder inhaler, which is then actuated, dispersing the powder. The resulting aerosol cloud is then drawn by vacuum from the device, where it is captured on a tared filter attached to the device mouthpiec. The amount of powder that reaches the filter constitutes the delivered dose. For example, for a 5 mag dry powder-contaiing blister pack placed into an inhalation device, if dispersion of the powder results in the recovery of 4 mg of powder on a tand filter as described above, then the ED for the dry powder composition is: 4 mg (dalivered dose)/5 rg (nominal dose) x 100= "Mass median diamesr" or "MMD" is a measure of mean particle size, since the powders of the invention are generally polydisperse (Le, consist of a range of particle sies) MD values as reported herein are determined by laser dfIaction, although any umber of commonly employed techniques can be used far measuring mean particle size.

"Mass median arodynamic diametr" or "MMAD" is a measure of the a dynamic size of a dispersed particle. The aerodynamic diameters used to decrbe an aerosolized powder in tm of its settling behavior, and is the diameter of a unit density sphere having the same settling velocity, generally in air, as the partile. The aemdynamic diamt encompasses particle shape, density and physical siM of a particle. As used herein, MIAD refers tothe midpoint or median of the aemurdynamic paricle si distibution of an aersolized powder determined by cascade impaction.

CA Detailed Description of the lnvention Tbic present invention is directed to the fonnulatio of cdry phospholipidci S inlralent cation based particulate comnpositiOI In penicular 1 the preset inven~tion is directed to the use of polyvalent cations in the mnufacture of phospholipid -conitainin.

00dispadble particulate compositions for pulmonary administraion to the respfrawiy ta IND for local Or sym ic therapy via serosolizaiiou and to thm particulate composition made o~h~y ThA invention is based, atient in paMt on the surprisin discovery of the beneficia aeroolization and stabiliaon propertes of phospholipd -contaiuinpmrculam o CCUMOSItirnR comprising a polyvalent cation. These unexpected benefits inclde a 0 dramatid= c Incras in the gel-o-liqid crystal phas transition temperacin (Tm) of the PArbiCUlaracomposiglion, improved dispersiilty Of such particulars compositions improved spreudabilkty of the particulars composition upon cat with lung epitlbelia therby allowing drugs to be more effectively distribuited to the mlg peripbery, and iWmred sAge stailirY of the pameicate cowpositions.

It is surprisingly unexpected that the addition of a very hygroscopic soft such as calciu chloride would amobillize a dry po~wder prune to mjoisture induce~d destabiliin as owe would expect tha&t the calcium chloride would readily pick up Water leading to pinhole aWegadon. However, this is not what in observed. In contrast addition of calciumn ions leads to a drmatc improvermt in te stability of the dry p~hosphlipidbaseci powder to hurmt. While, not being bound to any theory, it is believed that calcium ions are believed to intercalate the phospholipid membrane, thereby ittera odug directly with the negtively charged Portion of the zwiudonic headginup. The result of this intaciu is increased. de-hydration of the headgroup, are and condwaarion of the acyl-chain packing, 2ll of whichileads to inine d -odna stability of tbu phospholipida.

The polYvalent catio fOr use in thC Present invention is preferably a divgaent calioin including calcium magsin Zinc, iron, and the lima Accoring to the invention, thu polYvaen catio is prese= in an ainn effective to incrase thu Tm of the '0 phosphOlipid such that the pazticuaM exhibitm ts a flu which is greate tha its StwraUermmcune Ts by at least 20 0 C. prafarably at least 400 c he molrrto of polyvulen" cation to phoopbolipisi should be al least 0.0, preferably 0.05 2.0, and mos preerbl 0X5- 1.0. A molar ratio of polyVacl caepiehoii of about 0.50 is partcicuary proetend acwding to the present invention. Calcim n tihs p=rculaiy preftrrd polyvalet catio of therpeenA invetin ad is povded as caldcimlodrde.

o ~~In a broad sense, phospholipid suitae for use in the preset invention includ any of those known in the art.

0 According to a pre~ferred embodiment, the phospholipid is mnost preferbly aL Sated phcspbolip& According to a particularly preferred embodiment sated c-i S phosphaddylclaofnea are used as the phospholipid of the present invention. Peted acyl chain lengths are 16:0 and 1S: 0 (Le. palmitoyl and ataroyl). According to one embodiment 00 direcd to lung surfeetan compositions, the phosholipid can mak up to 90 to 99.9* wtw of tho composition. Suitable phospholipids a=c=ding to tbs aspec of the invention o ~include natural Or syntheic lung surhcrmn such as those comarnially available under thetrademarks ExoSurf, IntaSwT8Q (Ony, Inc.). Survant, CwMoSurt, and AJLEC For drug delivery purpoes wherein: actrve agent is included with the particulate composiion the c-i phospholipid coutent will bedetermined by the drug activity, rhe mode of delivery, and Other factors and Will Riely be in the rangefromnabour 20% to up to 99.9% wfw. Thus, drug9 loading can vaTY between abou 0. 1% and 80% w/y, preferably 5 70% w/w.

Accozrding to a preferred embodiUer, it has been found in the preen work d=a the Tm of cie phospholipid is critica in obtaininga phospholipid-based dry powder that both flow well and are readily dispersible from a dry powder inhaler (DPI). The Tin Of te Modified lipid mkropaxticles cen be manipated by varying t amt of polyvaleuc cations in the formulation.

]Phospholipids from both natural and synthetic source are compatible with the preen invention and may be. used in varyin canceufratins to form the strutual matriL Genealy compauvbe phospholipids comprise those tha have a gel to liquid crystal phase tusido greater than about 4&*C Preerably rth norported phospholipds ar relatively long chain C 16 -Q2) saturated lipids md mmr pefraby comprise saftured phMospoipids most preferbly saiwcatd pboapbtiylcholine having aty chain lengts of I" Oor 1.0 (palMitmYl and stearyj. Fxeuplwy phoapholipids usefu int the disclosed stailized pratons comprse, phboglycezieas suh as dipaRoypbosphatidylchuline, distuYlptosphatidylcholine, &arbchidoylphosphadlchogjje dgxbeb ~hphafidylahalin diphosphalidyl glycerol shorbchai phoephatidylcholi, 2 long-chair ~maMtard pbospbatdylezhaWlamjnes longchain satctd phosphadidylsazjnr, long-chain stinted phosphsddylglyoerol, long-hai BatMited hphylwosit~s In adtion to the phospliolipKd a co-aurcran or cmbinaions of sudacimus including toe use of one or mm in the liquid pins and o=A or =or assciated with t parmult OcwPoMdou aIre contrnnlars as bein wihin ftn cope of the invention. By '.ocino with or compiwe it is mum fti thdulare 0omposim may hompcrpcin o ~adsorb absorb, be coated with or be formed by the surfactat. Surfacmnrs tiude (N floorated and nonf9uornateri compounds and are selected from the group consstng of saturated =ud unsaRaretd lipids, nonfonic dawrgenm, nonianic black copoynjes, ionic surfictants and combinions thereof In those embodiment Comprising stabilized dispersions, such nlonfluorinsted surfawUmts will preferably be relatively insoluble in the sRuSnion medium. It should be emphasized rha4 in addition to the aformtjoued 00suwtants, Suitable fluorinaed surfaaats ate compatible with the teachings her=i and INDmay be used to provide the desired paepazrions.

Compatible nanionic detergets suitale as co-sfac a comprise. sorbitmit eat=r ncluding sorbitan trioleats (Span T1 85), sorbtan sesquioleae, sorhita monoolmate, o ~~sottitn onolaunate, polyoxyethyLeue (20) sorbita n~olaite, and polyoxyshtylen 0 (20J) -sorhjt raonaoleaw, oleyl polyoxyetbyleae ethier, sleay polyoxyethylene (2) ether, laurYl POlyoxyedaylene ether, glycerol esten, andi sucrose esters. OxThe suitable nonlionic detergents ca be asily identfied using MeCutcheon's Emulsifiers and Detergents (MePublishing Co., Glen Rock, New Jerey) which is incorporated herein in its entiret. Pzefeue block cooyi=r include diblock and Iriblock copolynmr of POlyoxyethylene and polyoxypropylene, including poiaxam 188 (FlurniaTh F-69), polOXanr 407 PhhroicTm F-127), and poloxum 338. Ionic murfatts such as sodium Sulfomucchmaft, and fatty acid soaps may also be utilized.

Other lipids including glycolipids, gangliside GML, sphhigomyelin, phosphalidic acid cardiolipirn lipids bearing polym chains such as polyethl= glycol, chitin, hyaurni acid, Or polYviuYlpyzrhdnr ipids bearing sunlnrd mono-, dh-, and poccha=ie ftty acids such as palmiti acd, stesia cid, and olde acd, cholesterl choletrOl ester, and choleaW~o hainiucinar may also be used in accordance with the teahing of this inverialLm It Will flt1IWe be appreciated tha the paniuculat composiftions according to the inventon maY, if desi*d conmin a combination of two or morm active ingrdint. The a4ub may be provided in combfntionm in a &in&l spedi= of partiat compositio or hidviual in eepram apecicsof p=nicuj coMostios ]For examp, two ortw actve aget MY be ineorporated int a single feed stock preparaton Md spry dried to provide. a single particalate compositon sped=e coMPrising a pluaity of acive agents.

Conversey, the indiviua acives could be added to separate stocks and spray dried Npmnely to Provide a pluralty Of PartiuhIW compoiGIo species with dirM= COOsgIon Ilk=c mndvidus species could be addied to the suqspsn mdium or dry 3S pwderd~p~mg cmp nE any desired proporton and Placed in the =mroo delavy sstm as described below. Fwth=,mas auded to above. the particulat o WxIPosilijan (wit or without an associated agent) may be combined with one or more Conventional a micronized drug) active or biosedive agents to provide the desired (1)dispersion stabiliy or powder dispersibility.

Based on tlhe foregoing, it will be appreciated by thos skilled in the art that a wide S variety of active agents may be inzorported in the disclosed particulate compositins Accordingy, the list of preferred active agents above is exemplary onl and not intended to 00 be limiti=. It will also be appreciated by those sidied in the sum that the proper amount of IND agen And the timing of the dosages may be desamidned for the patiulate compositions in aconance with already exisizg infonnation and without undue expmertadon.

(Nt1 In addition to the phosphoiipid and potyvalent cation, the Microparwicles of the o ~presen invention may also include a bioconsparible, preferably biodegradable polymer, 0 ca~polDymer. or blend or other combination thereof- In this respect usefu polymer cmps polylacrides. polylactide-glycolides, cyclodextina, polyacrylates, methylcellulose, Carbryerylcellulose, polyvinayl alcohols, polyanhydrides, polylactams polyvinyl pyrrolidonca, polysaceaides (denrans, starches. chitin, chitosan etc.) 1 byahnr acid Proteins, (albulmin, colaen, gelatin, em). Examples of polymeric resins dwn would be usfu for the preparation of perforated ink micopatcles include: sne-kutadiene, styree-isoprene, sryren-aCrylouiizie, ethylene-vinyl acetawe ethyleae-acrylate, ethyeneacrylc acid, ethylen thlaczylaitm, ethyleetyl acrylae vinyl-methyl methacrylac, acrylic acid-methyl mexbatrylaze. and vinyl chiloride-vinyl acetate. Those skilled in the Mr will appreciate the?, by selectig t appopriate polymers, the delivery efficienacy of the paxtcuWe compositions ader the stability of the dispaerso may be tailored to optimiz the effcdveness of the active or agent Besides t aforementioned polymer materials and aurfactants, it may be desirable to add other excipients to a partcuiate compition to iumve partice rgidity, production yield emnitted dose and deposition, shelftlifo and patient acomptnce Such optional eXCipint include, but ame not limited to: coloring agents taste masking agents, buffers, hygroscopic agents, 2Wmnddarns, and chemcal stahilirs. Farthe, viiou e=49-enu my be incorported in, oyr added to, the paricula te uix to prtvvide moanur and form to the pmficulaic composiin (iLe. micropbtea such as late particles). In this regard it Will be.

apprecited din the rigidi in component can ba remnoved using a pom.prodnction tstmhnque such as selective solvent exm~acdn Odte excpiets may include, but stunt limited to, carbohydrates including -diachnides and polysnecharides. For example, rmschides such ats dcflosc (sahydrous and monohyd=n. galactose, mnnitol, D)-ciuos;. morbitO] sorbose and the Mke disaccharides such as lactoe mairose, suorsa, trehalose. and *ae li0m o rds~acchzda such as raflose and the like; and other caibohydrates such as gtarces (N ~(hydroxycthylstarh), cyclodexnfts and xnmltodaxrins. Other excipicar suitable for Ume with the Present fflvention, including amino acids, ar known in the art such as thnse disclosed in WO 95n31479, WO 96,20K6 azd WO 96532149, Mlans of Carbohiydates and aminto acids are further held to be withina the scop of the present uvenioL Tht inclusion of both inorganic (eg. sodium chloride, etc.), organic acids and thei mite (e.g.

00 caroxylic acids and their salts such as sodium citrate, sodium ascarempeii IND gluonat, sodium gluconace trmethmne hydochloride, etze) and buffiers is als rm~tw h ncuimo salts and anic solids such as amnaiw cxboat, mmc~ul R~MW.a==iumchldde orcamphor are also cantnmplstad.

IND

o ~~Yet other preferrd eniboditents inclu de partculate compositions that My 0 c~~omprise, or may be coated with, charged species tha pmlung resdence tim at the point of contact or enhance peett through Muasa. For e=apie, anionic charges are known to favor mucoadhesion while eaic chaxges may Ibe used to associat the, farmed aariculate with nfegatvely charge bloactve agents such as geetc material- The charges may be imparted through the association or incarpcnuiou of polyanionic or polycad~oni Materials such as polyacrylic acids, polylysine, polylaccc aci ad chatos=n According to a pretennd embadinicor, the partculat compositions may be used in te form of dry powders or in the form of stabiized dispenions comprisig a nm-aqueou phase Accordingly, the disrsions or powders of the presentinvention may be used in cojunction With metered dose inbaler (MDIls), dry powder Whbalers (DPh), at==dar, nmbulizes or liquid dose inhllalion (LDI)) reebnique to proide for effective drug deivery. With repe= to inhalation therapes, those Sidied in the art winl appeciate thlat te hollow and porous micrup~ars Of the preset inventioin aM patcularly useful in DPXs Convemionald fPlf comprise powdered forumlatioms and devices where a meufd dos of msdicamen±, efkin alo or in a blend wit lacaos cnier particls is delivered its an aerosol of dry powder fo= ihaWlon Tb. niedicament is formulated in a way such tha it readily disperses into discret partcles with an AM] between 0.5 to 20 pm, preferably O.5-Spzrs and am further chncrcrid by ant aeroso partice size dkWaihtiton less than about 10 P. usmda amrdynmnoic diamen (IvtA) and preihsly less than 5.0 pm TIe eda aerodynamic diazrcr Ofte powder wil chamct risticnfy range fm about 0.5 -10 pMM pnferbly from ab=out 0-5.0 pm WmAD, umm preferably fim show 1.0 4.0 jim The powder is actaWe eiher by inspiaton or by s external delivery fore, suh as pMOsuried air. Exanples of Eft suitble for admbisttion Of the particulMt 12

VO

o compositions of the present invention are disclosed in U.S. Patent Nos. 5,740,794, C 5,785,049, 5,673,686, and 4,995,385 and PCT application nos. 00CV72904, 00/21594 and 01/00263. hereby incorporated in their entirety by reference. DPI fcamlations are typically packaged in single dose units such as those disclosed in the above mentioned patents or they employ reservoir systems capable of meteing multiple doses with manual tanfer of the dose to the device.

00 As discussed above, the stabilized dispersions disclosed herein may also be IN administered to the nasal or pulmonary air passages of a patient via aerosolization, such as with a metered doss inhaler. The use of such stabilized preparations provides for superior dose reproducibility and improved lung deposition as disclosed in WO 99/16422 hereby inoporared in its entirety by reference. MDTs are well known in the art and could easily Cl be employed for adminisration of the claimed dispersions without undue experimentation.

Breath activated MDIs, as well as those comprising other types of imprnovemeis which have been, or will be, developed are also compatible wirh the stabilized dispersions and present invention and, as such, are conterplated as being within the scope thereof.

However, it should be ephasizd that, in prefenred embodimens, the stabilized dispersions may be administered with an MDI using a number of different routes including, but not limited to, topical, nasal, pahnmoznay or orat Those skilled in the art will appreciate that, such routes are well known and that the dosing and administratiou procedures may be easily derived for the stabilized dispersions of the present invention.

Along with the aforementioned embodiments, the stabilized dispersions of the present invention may also be used in conjunction with nebuliwrs as disclosed in PCT WO 99/16420, the disclosure of which is hereby incoporated in its entiety by reference, in order to provide an aerosolizd medicamen that may be administered to the pulmanary air passages of a patient in need thet Nebulizers are well Inown in the art and could easily be employed far adninistmion of the claimed dispersions withot undue experimentio Breath activated nebulizer, as well as those comprising other types of improvements which have been, or will be, developed are also compatible with the stabilized dispersions and present invention and are contemalatd as being with in the scope theaot Alng with DP, MDIs and nebuizers, it will be appreciated that the stabilized dispersions of the present invention may be used in conjunction with liquid dose instillation or .DI techniques as disclosed in, for example WO 99/16421 hereby incorporated in its entiretry by re nce. Lqid dose instillation involves the direct adminiration of a mublized disperion to the lung. In this regar, diect pulamnary admkntion of bioasive compounds is pazriculay effecive in the matment of disorders especially whn= r vascuar circlaion of diseased pardions of a lJung reduces the effectiveness of LInvenous drug dellvety. W-ith respect to 1131 the stabilied dispezuons aM prerbly ci oseB31d in c Cction with partia liquid ventilation or total liqud venzllad. hMover dxc present in~vention may tblrtbecr compriseintroducing aL IterapeutWIcly benefia amount of a physiologiAly acceptable gas (such ats nitric oxide or oxyge) into the pharmucal oMLdiseson Prior to, during or following administraion Particulafly preferred embadinags of the invention incar spxv tkidd 00 hollow and porus particulate compositions as disclosed in WO 99/16419, hereby IND incorporated in its entirty by refeencea Such pardeculae compositias ompris particles o ~having a relatively thini porous wall defin~ing a lage internal void although, other void 0 0 contining or perforated swcm cmepatda well. In prferred embodizvan the oN p aatex couos n w'il further comprise an active agen= o Compositions according to the present invention typically yield powders with k denites less than 0.5 g/cin 3 cc 0.3 glcm preferably less 0.1 gfazn 3 and most preferbly less than 0.0-5 g/cm 3 By providing particles with vexy low bulk density, the- zimWin Powder M=s tha can be filled into a unit doase containe is reduced, which elinazes the need for canIer particles. That is, the relatively low densit of t powders of the pre.en invention Provides far the repr oducible Adnilnisu~atian of relatively low dose pharnaeuzcajcompounds. Moreover, the eliminatio of crier purtcle will potentay mkinie throt deposition and any N"g effect since the lage lactosle particles will impact the throat and upper awmay. due to their size.

ft will be appreiated tha the patiulars compositions disclosed hei comprise a smicterl Mat= ix thbits, de=e or comprie voids, pores, defets, hollows, spaces, inreati spaces, aperteres perfoations or holes no abolute shape (as opposed to the morphology) Of the peibiaed icotuu s geealy not critica and any overall confutation tha provides the desird charactmiatic is contmplatd as being withi the scope of t invention Accorigly, prferre e wbadimems comprise approxitmtey flphexig shapes Howeve, collapsed, deformed or fracamred parncleesar as In accord==c with the tahing heren the p=art5 composition will preferably beprovided in a "d trs Tha s the -rpartces willposses a moist.r content that allows the powder to remain chemcally aNd phy*Oaly Wtb& durin storage a am ienttemprte and easily dispesible As sudh, the misure commo of the uiosriclea is 4tically les thnu6S by welght, and preferably len 3% by weiht In some- instance the molars cant=n wil be as low as 1 by weight Of co=tft wil be wP~eated diat the moisti CIonte is, at least in part, dictated by dv,- forulation and is

VO

o controlled by the process conditions employed, e.g, inlet cemperamre, feed concentation, pxnp rare, and blowing agent type, concenration and post drying.

Reduction in bound water leads to significant improvements in the dispeibility and flowability of phospholipid based powders, leading to the potential far highly effcimt S delivery of powdered lung surfacants or particulate composition comprising active agent dispersed in the phospholipid. The improved dispczbility allows simple passive DPI 00 devices to be used to effectively deliver thbese powders.

Va Although the powder compositions are preferably used for inhalation therapies, the powders of the present invention can also be administered by other techniques Inown in the ID art. including, but not limited to itamnsclar, intravenous, intacheaL. inthraperitoneal o subcutaneous, and tranasdernL, either as dry powders, reconstituted powders, or Ss3uspensions.

As seen from the passages above, various components may b associated with, or inorporated in the partieate compositions of the present invention. Similarly, several techaiques may be used to provide particulates having the desired morphology a perforated or hollow/prous configuration) dispersibility and density. Among other methods, particulate compositions compatible with the instant invention may be formed by teehniqusa including spray drying, vacuum drying, solvent exzactim, emnuifition or lyophilization, and combinations thereof It will furthe be appreciated that the basic concepts of many of these teehliques are well known in the prior art and would not, in view of the teachings herein, require undue experimntation to adapt them so as to provide the desired particulate compositions.

While several procedures ae generally compatible with the present invention, particularly preferred embodiments typically comprise particulate compositions fomed by spray drying. As is well known. spray drying is a onastep process that converts a liqnid feed to a died particulate farm. With espect to pharmaceutical applications, it will be apreciated that spray drying has been used to provide powdered material far various administrative toues including inhalation. See, for example, Mt Sacchetti and MM. Van Oat iw Inhalation Aerosols: Physical ad Biological Basis fr Therapy, Hickey, ed.

MarcelD dekar, New York. 1996, which is incorporated harin by reference.

In geMerl, spray drying consists of brdngig together a highly dispersed liquid, and a safcint volme of hot air to produce evaporation and drying of the liquid droplets. The prpamrion to be spray dried or Al (or feed stock) can be any solution, course suspension, sdmy, colloidal dispersion or peste that may be atomnized using the seleed spray drying apps In preferred embodimts the feed stock will cmprise a colloidal system such a an emulsian, ravers emulsion, icromulsion, mlriple emulsion4 particulate dispersion, or slurry. Typically the ftd isspae itoac rnt f im it edarha ri ~evaporate the solvent and conveys the dried produc to a 'o'llector The spent air is then exhausted with the solvent Those skilled in the art wil appreiat that severa difhe types Of apparatus may be used to provide the desired product Fotr example, comnneial dspray dryers m n f cue by Rcia Ltd. or N=ro corp. w il effectvey ~roduce partices of 00It Will futher be appreciated that these spray dryems and specifically their IND may be idifid or dzed for specialized applications, ie. the o aimukanemous sraying Of two solutions using a double nozzle technique More specificaly, 0 a waterin-cil emulsion Cmn be amOi~zM fi on n~l a asbdl c=antmg an, a- IDadherent su~ch as ruanizol can be coomzed from a secod nozzle In othe case it maty o be desirable to push the feed solution thjough a custom designed nonzle using a high pressre liquid chrornaograpzy (HPLC) pump. Provided that mirsrcrscomprsin th correct Morphology ad/or composition are produaced t choice Of appaats is not citical and would be appammn to the silled artisan in view of dhe teachings herein.

Examples f spray drying Methods and sysa=m suitable for ualdug fte dry powders of the present invention are disclosed in U.S, Patent Nos. 6,077-543,6,051,256,6,001.336, 5,985,24, and 5,976,574, hereby incrpoaud in thei entiretY by referne WhAile the resuling spray-dried powdered particles typicay are aprxmateCly sphercal in shape, neary unifom in size and frequently are hollow, thre may besom devee Of irrgularity in shap depending upon the incorporated mdc eitand t spry drying conditions. In many instances dIispersion stability and dlisprsiblizy of the paruiculate Coznposiion appears to be inmoved if an inflatin agent (or blowing agent) is used in their production as diwclaed in WO 99116419 aired above. PaWrIlrly prefted crnbodirn=t comprise an emulsionl With the in!ftng agent as the dispere or continuous phse The inflting agentz iiPreferably dispersed with a smrctat solution, using, for instance, a conmaclly available mirfudzrat a presur of about 5000 to 15,000 psi.

This procea form an emuio81n, prefrMuty smhbilizd. by an incorporated swant,= tycally coMmsig subm droplets Of wate irmmisc~le blowing agent dispersed in an aqueous ConinuOm Phase. The fornuztiOn of such emulsion usig tisa and otin~ teamiqes recommon and well known to thoe in the at. The blowing agent is pWreably a tbwannrd Compcmi (e g. perfuorhexane, Pezfuoooctyl romide, pwluoozy etihaeh padro~MIdecaj perforobutyl efbrn) which vapofizes during the spzay-dsying proess, leaving behind generally hollow, poru eodnmcll ih miwopherg Otutalelquidblowing aetp icue is cb~ f'D ethyl WtaM~ alcoh10lS an hydroarboa Nitro gad carbon dioxid

VO

o gas are also contemplated as a suitable blowing agent Perflnorcoctyl ethane is paricularly preferred according to the invention.

0) Besides the aforementioned compounds, inorganic and organic substances which can be removed iunder reduced pressure by sublimation in a pos-production step ae also compatible with the instant invention. These sublimating compounds can be dissolved or dispersed as micronized crystals in the spray drying feed solution and include ammoniun c00 arbonate and camphor. Other compounds compatible with the present invention comprise IN rigidifying solid structures which can be dispersed in the feed solution or prepared in-situ.

These suctures are then extracted after the initial particle generation using a post- C 10 production solven extraction step. For example, latex particles can be dispersed and subsequently dried with other wall forming compounds, followed by extraction with a suitable solvent Although the particulate compositions are preferably fnormed using a blowing agent as described above, it will be appreciated that, in some instances, no additional blowing agent is required and an aqueous dispersion of the meadicaent andlor excipients and sBUTetaugs) are spray dried directly. In such cases, the formulation may be amenable to process conditions elevated temperatures) that may lead to the formation of hollow, relatively powus microparticles. Mreover. the medicament may possess special physicochemical properties high crystallinity, elevated maelting tempertre, surfce activity, etc.) that makes it particularly suitable for use in sech techniques.

Regardless of which blowing agent is ultimately selected, it has been found that compatible particulate compositions may be produced particularly efficiently using a Btichi mini spray drier (model B-191. Switzerdand). As will be appreciated by those skiled in the art, the inlet tempeature and the outlet temperature of the spray drier are not critical but will be of such a level to provide the desired particle size and to mmsultin a product that has the desired activity of the maicumut In this regard, the inlet and outlet temperatures ame adusted depending on the melting characteristics of the ftarmlatian components and the composition of the feed stock The inlet temperamre may thus be between 60 OC and 170 with the outlet temperatus of about 40 OC to 120 -C depending on the composition of the feed and the desired partiulae cabaracteristics. Preferably these temperauers will be 6 C to 120 OC for the inlet and from 60 C to 90 C for the outlet. The flow rate which is used in the spray drying equipment will generally be about 3 ml per minute to about 15 ml per minute. The atomizer air flow rae wil vary between values of 25 liters per mninute to about 50 liters per minute. Commdially available spray dryers are well K3 Moa to d dse in the art, and suitable settings for any particular dispersion can be readily detmitnd though standard empirical testing with due refrnce to the examples that folow. Of come, the conditions may be adjusted so as to preserve biological activty in ci lkrger moleCUzeg such S protMn a peptides.

Yfl ~Whatever components are selected, the first step in particulat production typcally cornPnses feed stock pmapantion If thz phospholipid based particle is intede to act: as a carrier for another active agent, the selected actiVe agent is disolved in a solvent, PrufeMbly water, to produce a cacenjed sadolon The polyvalent cation may beadde 00to the active aget solution or may be added to the phospholiid emulsion a. discussed IDbelow. The actve agent may also be dispesed dircty in the emulsion, particlarly in the o ~case of waWT insoluble agents. Altwunsively, the active agen may be incoporaMd in the 0 ~10 form of a solid partiulate disperson. The coinCenraflon of the actve agent usdis IDdependent: on the amnount of agen required in the final powder and th ezf~m of t o delivery device employed the, ine particle dlose for a lIC)I or PRM. As needed.

cosurfar-taucs such as poloxamer 188 or span So may be disersed int tsassouon Additionaly, excipiente such as sugar and starches can also be added.

In seted emboclinets a polYValenT caton-conaiin oi-in-water emlsion is 'hen frmed in a Separate vessel. The oil employrd is preferably a fuorocabon perflnorooctyl bromide, pert luoOcty ethante, perfluorudecalin) which is emulsifed with a phospholic For example, polyvalent cation and phospholii may be homogenized inl hot distiled wate 67Q using a suitable high shea mechanmical mixer ltra- Tuna model T-25 Mixer) at 000 rpm for 2 to5 rainutes Typically 5 to 25 g of fluozucarbon is added drcpwise to the dispersed aur~aan solutio while mixing. The inndng PoIyvaleus cadon-conuding perfinorocambon i watr emulsion Is the prvc~ued usin a high presnm homogen=e to reduce the partice size. Typicaiy the emulsion Is processd at 12,000 to 184)0 psi, 5 discret passes and kept at SO to SWC The active agent solutio and perfinzocabon emulsio are thai comed and fed into tespray dryen Typically the two preptration Will be scible as the emulsion will preferbly compris =n aquous ontious phase While the hiacte agen is solbilizd seprately for die purpose of t innatf discio it wil be apprciated thA in othe embdmthdo active agen May be solubilized (or dispersed) directly in the emulsion.

Tn such caes the active eulson is simply Spray dried without combinin a seprte Reive agetpropartimn Io any event, op=rtig conDiton such as inlM and outlet temperume ted mam, atomi2adcm presure flw raft of the dryig atr, and nozzle- cctgauon cmn be adjusted in acor~me vhfi th Mnf eguideline ini order to produc the required patice ia, a=d producton yield of the remuItng dry patile. ExM=lary meti. .r as follows a2n air 10let t'mp~nmm betwee 6C and l7rC; an air outlt between 40"IC to 120C, a

O

feed ate between 3 mi to about 15 min per minute; and an aspiration air flow of 300 lamin.

C and an atomization air flow rate between 25 to 50 min./ The selection of appropriate apparatbs and processing conditions are well within the purview of siled artian in view of the teachings herein and may be accomplished without undue experimentation. In any event, the use of these and substantilly equivalent methods provide for the formation of hollow porous aeodynamically light micropardcles with particle diameters appropriate for 0aerosol deposition into the lung. mierostrctures that are both hollow and porous, almost ND honeycombed or foam-like in appearance. In especially preferred embodiments the o particulate compositions comprise hollow, porous spray dried microparticles.

0 10 Along with spray drying particulate compositions useful in the present invention Smay be formed by lyophilization- Those skilled in the art will appreciate that lyophilizatian o is a freeze-drying process in which water is sublimed from the composition after it is frozen. The particular advantage associated with the lyophilization process is that biologicals and pharmaceudicals that are relatively unstable in an aqueous solution can be dried without elevated temperates (thereby eliminating the adverse thermal effects), and then stored in a dry state where there are few stability problems. With respect to the instant invention such techniques ame particularly compatible with the incorporation of peptides, proteins, genetic material and other natural and synthetic nmacromolecles in particulate compositioMs without compromising physiological activity. Methods for providing lyophilized particulate ae known to those of skill in the art and it would clearly not require undue experimentation to provide dispersion compatible micropartieles in accordance with the teachings hrtein. The lyophilized cake containing a fine foam-like structure can be mironied using techniques known in the art to provide 3 to 10pm sized particles. Accordingly, to the extent that lyophilization processes may be used to provide microparticles having the desired poroity and sie they are in conformance with the teachings herein and are expressly contemplated as being within the scope of the instant invention.

Beides the orem ntod techniques, the particulate compositions or particles of thepresent invention may also be fonned using a mathod where a feed solution (either enlsion or aqueous) containing wall forming agents is rapidly added to a reservoir of heated oil (e.g.perflubran or other high boig FC) under reduced presme. The water and volatile solvents of the feed solution rapidly boils and are evaporated. This process provides a perforated stnncre from the wall forning agents similar to puffed rice or poparz Preferably the wall forming agts are insoluble in the heated oiL The resulting particles can then sepmed fromthe heated oil using a filtering technique and subsequently dried Imdr vacuum.

Va o Additionally, dhe particulate compositions of the present invention may also be 0 fomned using a double emulsion method. In the double emulsion method the medicament is first dispersed in a polymer dissolved in an organic solvent (e.g methylene chloride, ethyl acetate) by sonication or homogenization This primary emulsion is then stabilized S by foming a multiple emulsion in a continuous aqueous phase containing an emalsifier such as polyvinylalcohol. Evaporation or extraction using conventional tectmiques and apparatus then removes the organic solvent The resulting mierospheres are washed, 00 IDfiltered and dried prior to combining them with an appropriae suspension mediam in aordance with the present minvention Whatever production method is ultimately selected for production of the particulate IDcompositions, the resulting powders have a number of advantageous properties that make O them partticunrlry compatible for use in devices for inhalation therapies. In partidenlar, the physical characteristics of the particulate compositions make them extremely effective for use in dry powder inhalers and in the formation of stabilized dispersions that may be used in conjunction withmetered dose inhalms, nebulizers and liquid dose instillation. As such.

the particulate compositions provide for the effective pulmonary administration of active agunta.

in order to maximize dispersibility dispersion stability and optimiza distribution upon administratdon, the mean geometric particle size of the parculate compositians is preferably about 0.5-50 Wn, more preferably 1-20 pm and most preferably .5-5 pm. It will be appreciated that large particles (ie. pater than 50 pm) may not be preferred in applications where a valve or small orifice is employed, since large particles tend to aggregate or separate from a suspension which could potentially clog the device. In especially preferred embodiments the mean geometric particle size (or diameter) of the partiulate compositians is less than 20 pm or less than 10 n Mom preferably the mean getmefrc diameter is less than about 7 pm or 5 pm, and even more preferably less than about 2.5 pKm Other preerd embodiments will comprise preparations wherein the mean gometri diameter of the particulate compositions is between about I pm and 5 pm. In especially prferd mbodiens the particulate compositions will comprise a powder of dry, hollow, porams microspheicaI shells of apro mately 1 to 10 pm or 1 to 5 pm in diame with shell ikessesn of approdrately 0.1 m to approximately 0.5 pm. It is a partialar advantage of the prOsent invention that the particulate concentration of the dispersions and structuml mtrix compaonents can be adjusted to opimize the delivery chrctristics of the selected particle size.

Althongh preerredmbdiments of the preset invention comprise powders and stabilized dispersions for use in phamceudle applications, it will be appreciated thatthe o partilate compositons and disclosed dispersions may be Used for a njrAbeofn phwmacmdapp~icataona. That is, the present invention provides particulfte (1)ccmposiatious which have a broad range of applications wher a Powder is sueded and/or aersolized- it particular, the present invention is especially effctive whe an S active or bioactive ingredient rmst be dissolved, suspended or solubiized as &ga as possile BY incrasin the surface area of the porous micropaxdecles or by incorporaticn 00 with suitable excipients as described herein, will result in an improvement in dispersibiity, IND Radler suspension stability, Ih this regard, rapd dispement appliations include, but ame Zc iitdt:zzwrg nts dsw deterents, food sweeteners candimen, spices.

mer fltaton etegethi ckening agents, folia fertlie, phytohormones, insect o munns insect repellents, pet repellents, pesticides, fugicides, disiufetmns, perftm=, deodoamuts, etc.

The foregoing description will be more fully understood with referenc to the followinug Examples. Such Examples, mr, however, merely representative of prefered methods Of practicing Wh present invention ad ShoUld not be read as limiting the scope of rht invention. Ekxmple I Effect of adldedqealimrn ion on the Tm of any-cirie hshlp The effect Of calclim ions on the gel-o-liquid caysta traniton temperatu r(m) Of spay-dried PhosphOOliid was invesigated. The resultinig powders were exaineud vftually for Powder flow Cbaracteris$cs, characterized for Tm using a difefrenzi scann calorinW (1)89.

Dry lung stmtai Particles comprising long-chain saturated phaphatidylhomn, 142s dipalmiboylphmsphadidylchouine, DPPC or &Moylphospha*Idycbon, DSPC and varyig aMunts of calciwn chloride were M=11 by an emulon-based spray-drying proces. Calcium levels were adjusted as M* ai equinalens relative to lie PC presnt wish Ca/PC (Mci/mel) 0 to 1, Acccatsgl, 19 gOf laturted phospburidylchlimn (Geuyie Carp, Cambridge, MA) and 0 to 0.18 g o! caldum cldd diyduTM (Fsher Scntfic Corp., Pltburgb, PA) wae dispersed in 4Fdhy 40 reL of hot deimnized water (T 60-70a C) using an tn-Tnrxu Oixer at SMOO-iOM00 rim for 2 to 5 mnue. 18 g of perfnoocl ethane, pFOE (F.

Tech, Totyo Japan) was then added dmopwise durig miumn at a rar of 2-s mI/in. Afm the addton was complete, the eMdfim a gmind iran addiRIperiod of not leIrm 4 miue at M 1,0- 12410 spin The resultg comwe mauon was then homogenized mute o lu:Igh 1)e-ssm wvith ain Avesrin C-5 homogcnza (Ottawa, 'Cana) at 8.00iovooO Psi for4 Ps, and at 18.000-20,000O psi for a final pass, The subtirdero fluorocarbon-jn..wagw emulsion was then spray-dried with a Rnchj B-191 NMi Spray-Drier (EawiL, Swiland). equipped with a modified. 2-fluid atomize S under the folowing conditions! inlet t=emrture =895 *C Outlet teXmpeaue 58-.61 C(J pUImip= 1.9 Ml Mim 4 am~iZe pressur 60-65 psig; atmmizer flow rate =30-35 CM. The 00aspiation flow (69-75%) was adjusxc4 to maintain an exha=s bag pressur 20)-21 mbar.

IND h pra y-dried phospholipid particles wee collcted Uugtesua uh or~ epator The volume-weighted muan geamemric iameter ('1MD) of the dfy phospholiparticles wsconfitnud by Iner lfiathuimi (SYrnParch lmk R1006, IN CWd-eef-c Gm n) andi rnged from 2-5 Am to 3 8 pm depending onA the The resuting dry phospholipid paradces were also Cha=raizd using a model 2920 DSC (TA Instrments) and by a Karl Flher moisture analyzer. Apprwdxely 0.5 to 2 mg dry powder was weighed in= alUmoimi sample pans and hrMeticaly seale. Each sample was analyzed using amodultd DSC n~de under the following cndioun equili-bratonat r. and 2 %Yrnin nimp to 15Q OC modulated 44- 1 0 C every 60 sec. The phooii Tm was defned. as the peak maxima of the frst endothermic nrnston Sim ea& reversing heat flaw diermgn For moisturie analysis, appxoxirnaey 50 mg powder was suspended in 1 niL of anhydrous diuthyfommIda The suspMsin was Oa ijecred diretly ito the ttrationi cell and the moistre conjm was derived. The residualostor content inthe.

spay-dried DSPC particles is shown in Table la, and was found to decrese as a funcuion of Or/PC male ratio. Talbles lb and Ie presemthde Tmn value for the various spray-dried PC particles as a function of the Ca/PC ratio. Hydratd DSPC and DPPC lipoons exhiit Tm value of 58 and 42 OC, respectively. Dramatic inceses in Tm were observed following sjny-dying, and withi increases in calccimntt The powder forreWions devoid of calcim ions ware highy cohesve, while die formulatons incorporatinig addcalcium freesfiowing powder!.

The present ==rrile iMhstUja taItc heydration statu of powdered phosphllpd gpmatfics gretly iflWU=ne their inkant -dnai and physicochemtical characterifftics 4 Le,, Tmand flow properties incues in phoepholipiri Tin am believed to dnecty Correlate wit hrm in tnnl stbilty, which could lead to an enhancement in log-amX storage 8tablit. In addton decread Moisture cont=n maLy also lead to VaO 0 01 Table Ia. Effect of Added Calcium on the Residual Moisture Coautnt of Spray.

Dried DSPC 0.25 1.I 0.51 1-4 Table Th. Effect of Added Calcium on the Tmn of Spray-dried DSPC CSJPCL olu) 58h() I 0 (hydrted) Th 42 o -63 0.25 I69 0.5 19 Effect -f Added ManSinMM Ins o M Tmof Surav-dMed Phospholiids Phospholipid particles stabilized with magnesium iv= were prepared by an emulsion-base spmaydryiug technMiqlue. The emoulion feedatock was prepaqred accrding to the- prOPedur described below. in tbe f=s step, OA.5g ofditaolhsbrycone DSPC and 0.126g magnesium chlorde bhydma qOsher Scentfi., Plttburgb PA) W= dispned in 4 1gof hot deionized wary (1'6 tow 70 0 C) USingan tJfra-Tuax Pily (model T-25) at 10,000 rpm for 2 min. 17g of perfiumcruotyl ethane was then added drop vw at a M Of approximately 1-2 mI/rein ducing mixing. Afta the fluorocrbon addiio was complete, the emulsion wa mixed fOr aR additoni perio Of not less than 4 adnmm ThM =Wsltin coars emulsio was twi pocmsd trIUigh N a hig peur (Avesti, Ottawja, Canada) at 18,00 Psi fOr 5 passs, to yield a submicro flUOroabon-in.

water emulsio stabizd Iby a mnwojar of DSPC The crmkioa was then spray-dried with a Euri model B-191 Mini Spray-De= under the foflowing spay candirions aspiraor69%, TC, outlet W=Wpfilw.8C feed pump=L.9 itt min-', and Aoniln flow rm=e33 cm Diffe m canig calo!dncj analysis oft dry particES meealed t Tm forft DSPC in the powder Was 880C as coream with 79T forwsa DSPC (Table Ib). This 23 Va foregoing exampe illUsaes the effct ions such as manesium have upon the C- thermodynamic propexties of dry phosphopid particl d) Eaplm ofSnra.Dried Lunf Surfactmnt fExoSuL~es Dry lung surfUctanx particles having the same components as Exosurt( (Gaxoo Wellcome, Reseazvh Triangle Park. NC) were manaond using a sny-cying process.

Va IN o achieve this end, the osmotc NaCI component of Bxosurf was replaced in one f=at io by CaC 2 Accordingly, I55 g of dipatltoylpbsphaidaycholkj and 0144z of calium chloride dihydrate or sodium chloride were dispersed in 50 mL of hot daionized ol water (T=60-70 0 C) using an Utlra-Turax T-25 mixer at 8,000-10,000 xpm for 2 min.

1 8Mg of perflucrooctyl ethane was then added dropwise dinng mixing at a rate of &min After the addition was complete, the emulsion was mixed for an additional period of not less than 4 miutes at 10,000-12,000 rpm The resufring coarse emulsion was th= hofmogenized wider high pressure with an Avestin C-5 homogenizer (Ottawa, Canada) at 8,000-10,00 psi for 4 passes, and at 18,00 20,000 psi for a fmal pass. In a separate fask 0-12 8 of Tyloxapol was dispersed in 10 g of hot deionimed water (=60-70 9C). The Tyloxapol dispersion was then decanted into a vial char contained 0.174 g of cetyl alcohol.

The vial was sealed and the caryl alcohol was dispersed by plaecing it in a sonication bath for minutes. The TylonpoYcezyi alcohol dispesion was added to the fluorocabon emlsio and mixed for min. The feed solution was than spry-dried with a Bucebi-i9i Mini Spray- Dier, equipped with a modified 2-fluid atomizer under the following conditions: inlet tcraue 85OC, oulet tempeiatnure W8*.61 0 C, pump 1.9 xDl min aton Pressre 60-65 psig, aroii flow rate 30-35 cm The apration flow (69-75%) was adjusred to maintain an exbanss bag presst 2D-21 mbar. A fra flowing white powder was collected using the standard BchA cyclone separator.

The pray-dried powders were manually filled into a proptietay blister package and boea-sealed The illing pmcedure was performed in a hmidity cfotled glove box (RHc All bliste packages were nmbercd, then weihed before and after fing to determine the amount of powder loaded The filled blister packages were stored in 4 desiccating box operated at 2% RH util use. Tb powders wee then tested for dzspesibility from a DPI described in U.S. 5,74(494- Emitted Dose testing of tha fnatins was assessed following USP guideline.

fr inhalation product. The actuated dose was collected using a SQL min7 flow rate hold fbi 2 seconds onto a typo AMR glans ter (Gelman, Ann Arbor, The emited dose was Va o calclated gravimerically kaowing the blistat weight, total blister filn weight, and net Cl change in filter weight Dry powder conmining sodium chloride exhibited poor powder flow, and did not aeroslize well. In contrast. the formuation in which calcium chloride was substituted for the sodium chloride yielded particles with good flow and excellent emitted dose character.

The diffrances in dispersibility between the two foundulations is ftbzher reflected in the 00 standanrd deviations of the emitted dose. The fozegoing exape illuastrates the ability of the VO present invention to alter and modulate the flow and emission properties of dry lipid O particles through the inclusion of calcium ions.

Table IL Fona ulation of Highly Dispersible Dry Powder Lung Surfactant NO Prparations 0 Dry Powder Ca/DSPC (mo/nmol) Emitted Dose Formulation xosurfr 0 10t33 "Exosurf' Calcium 0.5 87 3 Thermal Stability ofSprar-Drled Pho ho id Particles.

In the current example, the thermal stability of the spray-dried phospholipid particles prepared in example I were assessed. Acardihly 50 mg of powder was transferred into 20 mL glass vials and stored in a vacuum oven at 100C for I hour. The volume-weighted mass median diameters (OdD) for the powders ware deteramined using a SympaTech laser diffliction analyzer (ELOS 1006, Clansthal-Zellerfeld, Gemany) equipped with a RODOS type T4.1 vibrnig trough. Appmroximately 1-3 mg of powder was placed in the powder feeder, which was subsequently satomized through a laser heam using 1 bar of air pressut, 60 mbar of vacaum, 70% feed rate and 1.30 mm ftunel gap. Data was collected over aninterval of 0.4 s, with a 175pm focal length, triggered at 1% obscuration. Particle size distibnudions were determined using a Frfthhofer model. The volume-weighed ean aerodynamic diametrs (VMAD) for the powders were deterined with a model 8050 AerosizrLD particle si analysis system(Amherst Process am ents. Hadley, MA) equipped with an Aero-Sampler chamber. Apprwimately 0.2 mg of powder was loaded into a specially designed DPI testing apparatus. In this teat, the powder was aemsolized by actuating a propellant can containing WPA-134a through the loaded sample chamber. The design of this apparetu s s such to mimic actuation from an active DPI device and to a so me insight into powder flowabity or is ability to deaggregam are.atcl i distribuflona are prefead and ane beleved to be an ,fl ~Table ME depicts the thennal stability anid changes in partice siz (MM] and VMAD) for the variou spray-dried D)SC particles as a function of Cat DSPC (mollmol) ratio. The thermal stabiliy of the powder was found to inceas with increasig c-alcium Content SigMTnit structual. and particle size changes were observed for the, formulauion 00 ~devoid of calcium ionls, as evidenced by particle string and large increases in MUM and IND VMAD. The addition of small amonu of calcium ions (CaJDSC 0O2) resulung in a o SigniUpficant mPov'Ent im thieniai stablity of the phospholipd particles. More supng, 0 10 the apmay-dried phosphoiipid formoulation enrihed at CA/DSPC rafto of 0.5 completey INDtolerated the acceleraed storage conditions, as no significant changes had occuated as a o result of storage at IOO1C for 1 hlr The above examaple further ilustates the enhanced tbexnal stability of spray-dried phospholipid particles afforded by the inclusion of calcium Table MI. Aerosol clnracterlst ics of Spruy'Dxied IDSPC Powders failowing Storage at 100 OC for Ihour Ca/DSPC Ta Thermal Stability %Wo VMAD, MMD VMAUD

(!M

0 79 Sintrhin at 5Mill. 3-3 2.1 5.7 j 02 5 1 Sintering at 45 main 3.4 1.8 4.52.

Ruample, V Th e Mfet of &dIIed CacUm Tons on cM Stability.

The Objective of tis study was to examine te effec Added cailcium had on the PhYsWca stbilt Of lipid-base PMD! suspensons to moinn. Budeaunide, powder w=r prepared by ny-hying a fted soluttion compsed ofnmiwnizad drug paricles suspended in the aqueous phase Of a fluorccan,wuter emulsion. Accordingly, 0.8 g saruatod eg phosphadidyichonine (EpC-3, Lipoid KGIaiwghfn Geunny) was. dispesed in appmiiatayO n0iL hat deionxized wa~W cr WO Q) using an tJ~n-Turra mixer at W00 rpm for 2 to 5 minutes 20g of perfbron was, thou added drop wise during mxn. After the addition was complete, the emulsion was mxed fisr an additionial perid of not lhn inutes IU zemulthg ecom dio wfis hooeid n high prn wit -n Avestin C-S h pgnzr(Ouana Canad) at 1&=00 psi fr5 panes. The resltig subm===t emulsion was then combInedl with a second aqueous phas containig l- 3 g o ~~budaSOnide SUspendad in a solution comprising04g d-lactose manohydrate, and l34g CA ~calcium chloride dissolved in approximately 30g of deionized water The. combined solution 0 ~~WM than =md using an Ultra-Tuna mixe at 8000 rpm for 2 minutes to ensure dispersion ofte budesanide particles. Hollow porus budesonide. parties were pitpar~d by spraydrYing the dispersion with a B -191 Mifi Spray-Drier (Bllcht Flawil, Switzerland) nuder the following spray conditions: aspiradiou- 80%, inlet tempentureS5 0 outlet 00 terperanie5T7C, feed pumpe4S3 reL/rin, total air flow-- 22.4 SCFMZ Fls flowing white Va ~powders were collected at the cycoe separator. Scannin clectrot nmrscopic (SM) analysis showed the powder to be spherical and highly porous.

ApproximataLy 40mg of spray-dried budesonide particles were weighed iato 10 Id oN almrtmm cans, and crimp sealed (Pkmasol 2005/10, Pfaffikon, Switzeriand) with a 0 DP30/50 ACT 5 0 M'I metering valve (Valois of America. Greenwich, CT). The canisters war charged with 5 g HFA-134a. (DuPont, Wilmington. DE) propelan by Overpressue throgh the valve stem (Panmol 8805). To elucidate diffrence between. the budesonide f=omulfttions, Proellant preparations that were spikted with var~dng amounts of water (0 to 1100 PPM) were utilized. The amoun of the propellant in the can was determne by weighing the can before and after the GdL. The final powder cmicenuuion in propellant was wlw and forulated to provide a theoretical es-valve dose of lOIgg budesoide per Rctuation. Powder dispersion was achieved by placing the canisters in a soalcation bath for 15 Mein. 71m charged pMDIs were placed in quarantine for a period of 7 days at ambient condidans to allow the valve seals to seat For the purpose of this study tearosol fine particle friion, F 8 pm) was used to asses changes in suspension physical stability that had occured as at result of the Water acivty. The budesnid pMDIs were tsted using comonly acepted phaeicjprocedures. Th. methofid utilized was compliant with thes United State PhWOO~ia(lISP) procedure (Pbawacopeia Previews (1996) 22-306543098). After waie shots 20 doses from the test pMD~i were actuated int an Andersen mpactnr. The axcion from allithe plan, induction port and actuatr were pedonnd in closed conaier with an appropriate amount of methanolhwater 1, Wtv). The filber was insalled but not assyed bwausec thelyacylic binder intrbed with the anaysis.

Bndeaonide was quantified byr measuring the absorption at 245nm (lgeclean D37640 ,IeF t howmeter) and compared to an external standard curve with te extraction solvent a the blank The "PF was aiculare4d aceo±4hs to the USP mednl refewmed above.

The effect of added calcium 1ouW on fth physical stabiity of th= budasonide pMDha 31 is depicted in Figur I. The phyial stbilit of t. budesauld pMDIS was tad to minasm WOt inmasing calcium cancarie Suisi*l the tolerance of the budesunide pMDI o ~~~smmmesion to moistue increased from approximately 400 ppm to nearly 700 ppm by the 0 inclusion Of 4% calcium chloride into the fornmlion.

This example illustrates the enhacved stability of phosphbljWi4ned pMDI particle afforded by the presence of calcium ions. The ability of a pMDI formulation to tolerate increased levels Of moistur will lead to an enhancement in their long-term storae stability. The presenc of Water ftel Stuctr Changes, which cani led to formatin of li quid bridges between particles and/or zeciynallization of components and changes in IND surfrce chmczexistivs. Tite overall effect of Moisture ingress for suspension pMDIs leaS S Particle coarening and suspension instbility, all of wich ca lead to produc failume INDEump le V! o :Me Effect ofAdded CAteh Ions on Particle Mornholoitv.

The objetive Of this Study Was to examine the effec-t added calcium as upon the mmPhological character of spray-dried phospholipid particles. Scanning elect=o 'n1icographic (SE2,t) image of the spray-dried disrearoylphosphafidylchoiine particles prepared in eXaMPle I We=e taken. The powder were placed on douible sticky carbon graphite that was amxted an labeled aluminum stabs. The samples ware thensputr-coated with a 250-300 A layer of glpaaiu.Samples were examined on a scanning electron uneroscp Operated at aM accelerating voltage of 20 Key, and a probe curre= of 250 pAnps. Photornicrahs were digitally capeared at a 20,OOOX magnification.

The- efk~ct Of calciuIm ion concmBzadion an the morphology of spry-dried DSPC particle is fflunratd in Fit= IL Foanuladons contaiing calcimn ions bad a highlyV Porous sponge-like mflaed nuphology, wherea the nm= DSPC particles appeared melted and collapsecL The 1hollow Porus marphology is chAracrdred by powders that flow and MAersoiz wel wherea t collapsed MophIology minks in powders with poor flowabilt and dispersbiliry NO significan differece in morpholoy was obseved as a reslt of calcium ion c nnUrion although the CaIPSP& 0.25 formuaton exhiited so=~ deg=e of melted characr as welt. The decreased sensiivity of the powders with highr calcium Con to eligand partcle fusion is liWY t Mih of the incrased bi values tha alw fir the powders to expeienc at higher drying tempgran while mainmining the lipidS in the gel state. The signifcant incrases in TM obseved (Exaple!1) lead to greate iftliity= pra-dr mndw~eof theme Pardele and a uignificamly gre liklMood Of achiein deird partcle ruorphologies which are dependen on drying rates Exmple VIU EPnartion of Snrsy-Direkfludesonicie Particles Hollow porou0s budesoaidc particles were pre-pard by a twa-step process. in the firt Step, 54Mg of budesonlde (VincheM, Cbathan and 0.775g of DSPC were 00dissolved in 2 mil of cbJorofornrmeVarnI The chloyOfOnijcti* 50 1 was then to obtain a thin film of the phosphoipidlstezid mixtre The phosphalipI&sterojd misre was thent dispersed in 30.5Sg of hot deionized water (T o 70C) using an Uftra-Turra Mixer (mdel T-25) at 8000 rpm for 2 to 5 minutes. 12.8g of INOpezfluorooctyl ethane was then added dcipwise during mixing Afte the addition was complete, the emlsio wa med for an additional period of not les than 4 minaret The CORM emulsion was then passed through a high pressmre homongenizer (Avestin, Ottawa, Canada) at 18.000 psi for 5 passes. The resulting submiercc fluorocarbon-in-waxer with stmi solubiiza in the lipid monoayer sunding the droplets was utilized as the k4edstock in fibr t he secn d UtpP, spray-dryin an a B3-191 Mini S pray-Drier (BilUhc Flawil, Switzerland). Calcium chloride (0 cr 0-65 rmg) was added in 2-5g of water to the fluorocarbon-in-watmr emulsion immiediately prior to spray drying. The. following spray conditions were employed: asphionlc-70o, ine mperaturuS5 0 C, ouder telnperahrer-6oac, feed pump=1I.9 mL mWi amoziznr ssM r-65 psg atomize flow rsb--30-3s CM~ The aspiration flow (69-75%) was adjusted to niutain an exhaust bag presure of 30-31 mbar. Pree flowing White powders were collected Using a standard Cyclon Seiaratr.

The resultig dty budasonjde. particles were characterized usig DSC. Ea&h sampl was analyzed in a modulated DSC mode fuert follow*n condidios± equifiration at aM 2 QC/min ramp to 150 0 C modulated IOC every 60 we- The phospholipid Tm was defined as the peak maxima of te fit euduxher transition from each revering hear flow thanrog TU pbospholipw Tmn for DSPC partie without added calcim is 79-2.

Tim addit Of calcim ions in the budacovjde to nulation inmaed the Tm to 98C. In ad&ideUILthe powder ffonlatiaua devoid of calcium. bad a cohesive flow charctar as TLIM aerosol characticsf of the calCiurm contaiig farmulation was examined in see passiw dry pwd inhWe device (clips@, Turbospin@, CIpla Rotabalen, Glxo, Rt fo and Zioion PlowCaps). The emifted dose was deterned gmvimWricy at comforta inhalato flow rate (peek flwnt 20.62 L/nin depending on tha resstnc of ia device), amd at a forcd Inhalato flownt (peak flow rare 37-90 M=ui). tine Contoymble inbasla aon flow coaditiana the rang of emitted doses was between 89 and!96% Cl with a menemitted dose of 9417. Under foited Inhalation flow, the emitted dose varied between 94 Find 103%. wiTh a mean emhwtd dose oa 99 The- fct tMal nmldple devices with high and low redsstanc mi able to effectively disperse the powders mome or less indepeudgmt S Of inspawr f ow raz speaks volumes to the disperdibiliry of the calcium containing budesoncide powder tested.

00 The above example further fiusttes the ability of the present powder engineerig IND teelmology to effixtively mzodulate the Toitiroug foruation Change= Increaed (Fms) are o ~~desire as they Offe= indicate increased physical stability and improved powder disprsiblliy.

0 EMwmple VIU kflvid Snes nof SuPraybDried RSP!C Pa~cJdW on an AixrWAter btorface The rapid spreading chameeistucs of the disclosed spray-dried phcspholipidbased partic-les at the air/ware intefac ar illustaed in ig. fi. S urfac team esm were made on a Kruss K12 tensiomee at 25 0 C using the Wimemy plate technique. To me-stwe surface tension, 20 mL of DI water or DSPC lposome dispersion was placed in the thermostatic- beaker. The plainum plate was tared in die air and ten dipped into the liquid ad moved into the interface, afier whi& chs~c were taken. Fur spry-drie DSPIC particle analysi wm nrta for DI water were made and confirrmed to be 72*-t1 iiiMl TrjAISWr and pla were re-cleaned if the sufae tendon was not within 2S expectation. ARoxinrey 0OS nag of dry DSPC crystal was sprinked. careizly onto the surfac while- the plat Was dipped into ib DI war Masm ents were starte inmledinelYr after the Powder was added. Cane was taken to ensue dry powder did not adsorb to the Plate, Mt W wer eA= sed if any powder had contacied the plate surface The eQuillb~riumxmrrac tension of distearoylphospbaiidycbointe, tDSPC) is ca. 22 MNl'hn. Aqueou based DSPC liposomues adsorbe very slowly at t akwaner intrfac as evidentced by the fiat that after 240 se., the surfac tension bas not boon signiflantly reduced. The slow adsorption for iposoes is due to the slow moleular diffusion of DSPC trough the water phase result from its exteMey low solhlity in water.

Surprisingly, the adsorption of DSPC in tie form of spzuydded, DSP particles is very fast reducing the smfie tension to equilhtjnrkm values within a few se=&ds Moreover the inclusion Of calcium ions had no effet on the_ spreading of surfactant properties of the DSC particlns. This rapd spreading ad reduct of sufmc tnsi is indicative of what would likey occu upon contctn te 'pray-dried phospholpid particles with a wetted Va pUlmonary membrane. Specifically, the present example provides a model forte effective delivezy of synthetic lung surfactars and drugs to the lung.

C)

Earation ofNico ine Bit xt re Partices for pMDLs bY ny4)rying 00 -Hollow porous nicotine bitartiar particles were prepared by a spray-drying technique with 4 -191 Mini SPray-Dutie (Echi, lawil, Switzerland) under the following spray conditions: aspiration 80%, inlet temperatre: 8T*C; outlet temperatre: 56C; feed pump: 2.3 mLdUin; air flow- 28 SCFKv The feed solution was prepared by miing two solutios A and B immediately prior o spray drying.

0~ Solution A: 5.2g of hot water 50-60 0 C) was used to dissolve O.

6 0 g of nicotine bitirrate (Sigma Chemicals, St- Louis MO), 0.127g d-l lactose (Sigma Chemicals, St Lois MO), and 90 mg calcium chloride dihydrate (Fishr Scientific, Fair Lawn, NI).

Solution B: A fluorocarbon-in-wamr emulsion stabilized by phospholipid was prepared in the following manner. The phospholipid. 0.69g SPC-3 (Lipoid KG, Ludwigbafen. Germany) was dispened in 29g of hot deionized water (T 60 to 70 0

C)

uing an UhIra-Turax mier (model T-2) at 8000 rpm for 2 minutes G 60-70 0 30.2g of perfluarooctyl ethane (F-Tech, Japan) was added dropwiss dining mixing After the fluoocrbon Was added, the emulsion was mixed for a peiod of not less than 5 minutes at 10000 ipm The resulting coarse mnlsion was then passed trough a high pressure haoenizer (Avectin, Ottawa, Canada) at 18,000 psi for 5 paws.

Solutions A and 1l were combined and fed into the spray-dryer under the conditions described above. A fre flowing white powder was collected at the cyclone separatr The geometic diameter of the nicotine bkaXsx particles was conired by laser dffacdon (Syinpatuh Halos H006M C ushal"-eerfex, Gennany), where a Vaam weighted mean diameter (VMD) of 2 .E0am was fond. Scnning electron umeros=ouy (SEM) analysis showed the powdm to be spherical and porou. Differential Weanfing cabormetry aalysis of the dry partcls (TA hs u t) veAied the Tmfor the nime bitarab= in the powder to be 62C, which is similar to what is observed fo spray- &d neat nImdift o Jxamule X Hollow porous nicotine bitarars particles were prepared by a spray-dring techique with a B-191 Mini Spray-Drier (BlIchi, Flawil, Switzerland) under the following 00 spray conditions. aspiration: 80%, inlet tmpeatur: 83-C; outlet temperdture: 57rC; feed 1-1 pump, 2-3 mnxun; total air flow.- 22.4 SCUM.

o A fi mcurbon-in-ware emulsion stabilized by phospholipid was first prepared.

010 The phospholipid, 0.4g SPC-3 Upoid KG, Lndwigshafea Germany), washo geid in 30g of hot deionized waxer (T1= 60 to 70t) using an Ullra-Tunx mixer (mdel T7-25) o at 8000tWrpm for 2(T=60-70'Q2. 15g of perfinoroctyl athane (F-Tech, Japan) was added dropwse at a rare of approxiately 1-2 min during mixing. After the fluorcarbon was added, te emulsion was mixed for a period of not Less than 4 minuteS. Tim resulting coarse emulsio Was then processed rbwugh a high presure homgenizr (Avesrn, Ottawa, Canad) at 18,000 P4 forS5 passes.

The emualsioin was decanted into a beake contaiig 8 mg sodium phasphate zuonobasic (Speettmn Chemcals, Gardena. CA) Ws 90 mg calcium chloride dihydrate (Fisher Scientific, Fat Lawn, NJ). The emulson was alwed to t for approimately 5 nziii The emulsion was then decanted into a beaker contaiing 0.2 2 nicotine bitnazte (Sigma Chemicals, SL. Louis MO) and was sdnved for 5 minutes. Thke fEed solution was fed into the sp'ay-dryer nder the conditions descried above. A free flowing white Powder Was collectod at the cyclone. separar. The nicotine bitarna particles had a volume-weighnets mean aerodynamic diameter, of 1.47 gsm as detemlined by a time-of-flight analytical mothod (Acrosirci Amhers Processlmnmns Amhers MA). 71e gemnetric diameter of the nicote bitaxraaaB particles was detemnemd by laser diffiuctim (Syrupatech Halos MOM06 OaUnhMadeljd, Ciennany), when a volume weightd man diameter (VIED) of 2-95A= waa found. Scanning eleamronmoscopy (SM) amaism showed die. powders to be spherical and highy porouL DidtiAl sanning calduuzzry analyis of the dry parties (TA bBUifents) revealed the Tm for the, nicotine bftrzzut in the powder, was arnoxinely 8S'C This fMOging exmple IMutates the abilt of the presen powder engieering technoog to effetvely, zmdular t Tmtbrogh fomulatio chan

VO

o Example XI The preparation of lung sufactant powders with and without the use of blowing agents was investigated. The resultant powders were characterized as to aerosol properties.

Preparation of powders 00 IN The annex solutions were prepared by mixing calcium or sodium chloride cetyl O alcohol, tyloxapol (Sigma), and Infasurf (ONY Inc.) as desctbed in Table IV in a 20m1l SO10 glass vial to which was added an amount of hot deionized water (70' C) (appximately ci 0.54 g of sodium chloride were used instead of calcirum chloride in lots 1843-HS-03 and o 04). The mixtue was vrtexed wntil all solids were fully dissolved. One lot, 1843-HS-04 used 200 ml ethanol as a solvent.

The emulsions were prepared by adding DPFC into a beaker to which was added an amount of 70 0 C deionized water. The mixure was mixed in a mixer on low speed for approximately 2-3 minutes. When a blowing agent was used, PPOE was weighed out into a small flask and added dropwise into the DPPQwar mixture. The PFOE was added slowly.

over the course of 1-2 mintas and the mixure was then allowed to continue mixing for an additidonal 1-2 minutes. Emulsion details are listed in Table IV.

When a blowing agent was used, the DPPCwater/PFOE mixture was the.

immediately removed from the mixer and run through a homogenizer four times at 10,000 13,000 psiL The sample was then rnm though a homogenizer a fifth time at 13,000 17,000 psi.

The annex solution was then added to the DPPC/water or DPPCwatrOPPOE emlsion with continued stiring on a hot plate set to the lowest temperatu The mixtres were kept at approximately 50" C during spray drying, which was done at the conditions listed in Table V.

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IND Aftm thff addtion is complet 7 the emulsion is mixed for an addtional period. of not less thant o ~~~minu~es at 10,000 rpm Te reulting coare emusion is then homogenizc under high pruesr ci with an AveSdin C-5 homogenizer (Ottawa, Canada) at 19,000 psi for$5 discete passes men emulsion is rranftnd to the Potnt Molecule Laboratory for Leuprolide Acetate addition and sPlay drying.

Table Vt.

Leurlijde ActtHMUlioCoustp &Mn-1sion Components AnMOMt solid aio 00N Aaosoi Data Depositon analysi is pedbxmeA using a amid-stage Jiqid impinger (iv=Si) Mm apparatus consists of for caacunemt stages and a ten iu each containing an aliquot of aVRpprIn solven for Leuprlit Acetae anialysi. Deposiliw and emio data is repored in Table Vffbelow.

Lec roldeTable V I LS~-ru-ldeAcetate Aerool Dais Lo~t X=216 96% 00 Dose IND- 2.40 o 54-ilter70% o .4 o P~Fh Feed Solution Prepartion A single feed solution is prepaxed under defined condition. The feed soluion is compxiscd of parathyroid hormone in the aqueous phase of a flaoroco-in-water emulsicit The emulsion composition Us lstd in Table VMI below. Accordngly, DSPC and cacium Chloride dihydute are dispered in approximnately 4OrnL SWF[ 70 C) uimng an Ulta- Turn T-50 mixer ar 8000rpM for 2 to 5 nbnmes. The perfinroactylethae is then added drop vise- duringl mixing After the add ihia is complete, the emulsion Is mixed for an hddironal period Of not less than 5 minuts at 10,000 rpm. The resuking coarse enmison is then homogenized under bigh pressure withian Avestin C-5 homogenizer (Ottawa, Canadia) at 19,000 ps for 5 &=sreM passes. The active OrM is added to the eWmuo and subseqently spry dried after mixing for aperiod of not less than l0minmas.

TabLuVlfl I PeMratyod Zftmo-M Emulsio Comog4on Gn~nC~noet Amunt (MMn) suolid!; 0.0757-5* 28 NA 40 JNA An jsnrjz -L 0 2S Deposizon ansi is perfommed usig an Anderuon Cascade Impacto. The appaub consist of seven "cnmt stages and a termlinal Liter. Aausol deposifion is masured VWM6milyand is rMpoed in Table Is below.

?arat-hyroid flpjone Aerosol Data Lm2193- E&MPsAIxr Flow Rim 30L m PriMrda f Mrd Dose 00 TnholAw 2.67 INDj~ 59% coitnni Nttie BitaStnte Partc'e o 10 5(1mg of nicoine bfttraprdclas prepared in Examples Zand X wer weighed into 10 ml aluminum cans, crim sealed a DP350S RCU-2Ocs SWp valve (ValoisofAeia Greewich, C7) and charged with HFA-134a (Dueozi Wilmigton DIE) popluat by ci OV~m PrmnMP through the Stem A Pamasol (Pfaffiko, Switzeland) model 200)5 small scale production plant complete with a model 2008 propellan pump was used for this pupose The amount Of The propellant in the can wus detened by weighing t can before and after the fill Thefinl Powder Cancenrion in propellant was 0.5 9 wfw and fomulated to provide a approximte emitted dose of 110 gnicotine bitmrtmte.

Enmule XV The MDIs were tested usig co mony accepted pharinaceuca procedure. The method utilized was coroplia with the United Stat Pharmacopela (IJSP) procdre (macopea Prevews (1996) 22:3065-3098) incorortd herin by refernc. Aftr waste. shot, 20 doses fain the test pMDrs wete=Maied into =n And&rsen rpato.

EMWm~mW.The exfran from all the phlatdction port, m~d fa= were padbfod in closed canm=z with an apprate amof mntbwiotwater 1, Wv).

The fiter was istlled but not assayed, becaus the palyacryli binder intferud with the anaysis The :mass balanc and patcle aiz &dlutibdon trends indicated that the depositio on the filter was negliibly sal] Qumtwon rocedre NicoUMn bitarwar was qandrtad by mensig the absorption. at 258jnm (Becbman DU640) spectrphoomgmy-) and compared to an erna mandmd CUrV_ wi t extaclion Woleur as t blank.

For ealch MD= tem of the drug in the stm (componet niarm indction pmr and plats (047) nA qunified. as descrbed abov& ThU Va Fme Particle Dose and Fine Particle Fraction was calculated according to the US? method referenced above. Throat deposition was defined as the mass of drug found in the induction port and on plates 0 and 1. The mean mass aerodynamic diamers MMAD) and geometric standard diameters (GSD) were evaluated by fitting the expedmmwal cnxnlatin function with S log-ormal distributon by using rwo-parameter fitting routine. The results of these 00 expeiments ar presented in subsequent examples.

Va oo 01 INO en Cs;ngC~pad k* _Warto Realts-for Nicofing ]3ftyzft aMUF-mm~at The rests of the cascade impactor rests for the nicotie bitartrate pMDs pepaed accorfing To Example XIV are shown below in Table X Table X Nicotne Biiarmie Das NQLAD Hue particle Pine Parifiel Dose, (CSD3 fration. Ag Am 9b Nicotin/SPC3/Cacl 3.6 70 74 Lacws eO ModndSPCc3/Cao2 3.0 73 Both MDI prepaations were obsed by visual inspection to have excellent sspension stabilty, where little or no creaming or sedimentation occurred over 1 hour. The lacOse containing formnladons had a slightly larger MMAX and lower FPF and D a compaed with the sodium phosphate fonmtlaon The redaction in aerosol perface for the lactose formulation could be due to inaeased water content as evidenced in the reduced The invention has now been deSihed in detail for purposes of clarity and 23 nderstanding However, it wil be appreciated that certain changes and vadificaum may be pacticed within the scope of the appended claims.

40 o In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word C) "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e.

L to specify the presence of the stated features but not to preclude the presence or addition of c 5 further features in various embodiments of the invention.

It will be clearly understood that, although a number of prior art publications are 00 referred to herein, this reference does not constitute an admission that any of these documents 0 forms part of the common general knowledge in the art, in Australia or in any other country.

O 1 K:\I~ara\ee\Seci\T 3tl03..dc 2312/06

Claims (32)

1. A particulate composition for delivery to the pulmonary system, the ri composition comprising: C 5 particles comprising an active agent, a saturated phospholipid and a polyvalent cation, wherein the molar ratio of polyvalent cation to phospholipid is at least 0.05 and is 00 sufficiently high to increase the gel-to-liquid crystal transition temperature of the particles \0 compared to particles without the polyvalent cation. 0 0 S 10 2. A particulate composition according to claim 1 wherein said gel-to-liquid o crystal transition temperature is greater than room temperature by at least 20 0 C.

3. A particulate composition according to claim 2 wherein said gel-to-liquid crystal transition temperature is greater than room temperature by at least

4. A particulate composition according to claim I further comprising a surfactant selected from the group consisting of nonionic detergents, nonionic block copolymers. ionic surfactants and combinations thereof.

5. A particulate composition according to claim 4 wherein the surfactant is selected from the group consisting of sorbitan esters, cthoxylated sorbitan esters, fatty acids, salts, sugar esters, ethylene oxides, and combinations thereof.

6. A particulate composition according to claim 1 wherein the saturaled phospholipid comprises a saturated phosphatidylcholine.

7. A particulate composition according to claim 6 wherein the saturated phosphatidylcholine comprises dipalmitoylphosphatidylcholine or distearoylphosphatidylcholine.

8. A particulate composition according to claim 6 wherein the saturated phospholipid is a zwitterionic phospholipid.

9. A particulate composition according to claim I wherein the polyvalent cation is a divalent cation. H:\MaraR\Keep\Speci\Pf003E.doc 23/02/06 42 A particulate composition according to claim 9 wherein the divalent cation is selected from the group consisting of calcium, magnesium and zinc.

11. A particulate composition according to claim 9 wherein the molar ratio of divalent cation to saturated phospholipid is 0.05

12. A particulate composition according to claim 8 wherein the molar ratio of divalent cation to saturated phospholipid is 0.25

13. A particulate composition according to claim 1 I wherein the divalent cation is calcium.

14. A particulate composition according to claim 12 wherein the molar ratio of calcium to saturated phospholipid is about 0.50. A particulate composition according to claim I wherein the saturated phospholipid comprises a natural or synthetic lung surfactant.

16. A particulate composition according to claim 1 comprising 0.1 80 w/w of the active agent.

17. A particulate composition according to claim 1 wherein the active agent is selected from the group consisting of nicotine, human growth hormone, parathyroid hormone, leuprolide, budesonide, tobramycin, albuterol, insulin, interferon alpha, interfcron beta, amphotericin, fluticasone, salmeterol, formoterol, and salts thereof.

18. A particulate composition according to claim 1 further comprising a polymer selected from the group consisting of polysaccharides, polyvinyl alcohol, polyvinyl pyrrolidonc. polylactides, polyglycolides, polyethylene glycol, and mixtures thereof.

19. at least one of: A particulate composition according to claim I wherein the particles comprise a mass median diameter of less than 20 microns; and an aerodynamic diameter of less than 10 microns. i \-arbR\KeepSpec: \FC)o3Sdc. 23/C2/06 43 O C, 20. A particulate composition according to claim 19 wherein the mass median Sdiameter is within 0.5 5 microns. 5 21. A particulate composition according to claim 19 wherein the aerodynamic diameter is within 0.5 5 microns. 00 ID

22. A particulate composition according to claim I wherein the particles are hollow O Sand porous. \O to S23. A particulate composition according to claim t comprising an emitted dose of at least

24. A particulate composition according to claim 1 further comprising a non- aqueous suspension medium. A particulate composition according to claim 1 fitrther comprising an excipient selected from the group consisting of amino acids, carbohydrates, inorganic salts, organic salts. carboxylic acids, and mixtures thereof

26. A particulate composition according to claim 25 wherein the excipient is selected from the group consisting of hydrophobic amino acids, monosaccharides, disaccharides, polysaccharides, sodium citrate, citric acid, ammonium carbonate, ammonium acetate, and ammonium chloride.

27. A particulate composition according to claim I wherein the bulk density of the particulate composition is less than 0.5 g/cm.

28. A particulate composition according to claim 27 wherein the bulk density of the particulate composition is less than 0.05 g/cm'.

29. A particulate composition comprising: particles comprising an active agent, a saturated phospholipid and a polyvalent cation, wherein the molar ratio of polyvalent cation to saturated phospholipid is at least 0.05 and wherein the composition has a gel-to-liquid transition temperature at least 20'C higher than H:\MaraP\Keep\Speci\P6003j.doc 23/02/06 44 VO O room temperature. O c' C) 30. A particulate composition for delivery to the pulmonary system, the Scomposition comprising porous particles comprising: C 5 20 99.9% of a saturated phospholipid; a polyvalent cation, the molar ratio of polyvalent cation to saturated 00 phospholipid is at least 0.05; and 0.1 I- 80% active agent. O O 0 10 31. A particulate composition comprising: O 0 particles comprising a structural matrix comprising a saturated phospholipid and a polyvalent cation, wherein the molar ratio of polyvalent cation to saturated phospholipid is at least 0.05 and is sufficiently high to increase the gel-to-liquid crystal transition temperature of the particles compared to particles without the polyvalent cation, and wherein the particles further comprise an active agent.

32. A particulate composition for delivery to the pulmonary system, the composition comprising: particles comprising an active agent, a saturated phospholipid and a polyvalent cation, wherein the molar ratio of polyvalent cation to saturated phospholipid is at least 0.05 and less than 2, whereby the gel-to-liquid crystal transition temperature of the particles is higher than particles without the polyvalent cation.

33. A method of making a temperature stable particulate composition for delivery to the pulmonary system, the method comprising: forming a feedstock comprising a saturated phospholipid emulsion and an active agent; adding a polyvalent cation to the feedstock in an amount sufficient to provide a molar ratio of polyvalent cation to saturated phospholipid in the feedstock that is at least 0.05 and less than 2; and drying the polyvalent cation containing feedstock to form porous particles having a gel-to-liquid crystal transition temperature that is higher than a storage temperature ofthe porous particles by at least about 200 C. H:\MaraP\Keep\Speci\P60036.doc 23/02/06 45 O 34. A method according to claim 33 wherein comprises adding the polyvalent 0 CN cation to the feedstock in an amount sufficient to provide a molar ratio of polyvalent cation to saturated phospholipid in the feedstock that is from 0.25 to I.

35. A method according to claim 33 wherein the polyvalent cation is a divalent cation. 00

36. A method according to claim 35 wherein the divalent cation is selected from o the group consisting of calcium, magnesium and zinc. Va S37. A method according to claim 35 wherein the divalent cation is calcium.

38. A method according to claim 33 further comprising adding to the feedstock, a surfactant selected comprising nonionic detergents, nonionic block copolymers, ionic surfactants and combinations thereof.

39. A method according to claim 33 further comprising adding to the feedstock a polymer selected from the group consisting of polysaccharids, polyvinyl alcohol, polyvinyl pyrrolidonc polylactides, polyglycolides, polyethylene glycol, and mixtures thereof. A method according to claim 33 comprising drying the polyvalent ion comprising feedstock wherein the particles have a mass median diameter of less than microns and an aerodynamic diameter of less than 10 microns.

41. A method according to claim 33 comprising adding an excipient to the feedstock, the excipient comprising amino acids, carbohydrates, inorganic salts, organic salts, carboxylic acids, and mixtures thereof.

42. A method according to claim 33 comprising drying the polyvalent cation comprising feedstock to provide a bulk density of the porous particles that is less than g/cm3.

43. A method of delivering an active agent to a patient in need thereof, the method comprising: k: \MaraR\Keep\SPEc:\P6OC3--.oc 23/02/O6 46 administering to the respiratory tract of the patient an effective amount of particles comprising an active agent, a saturated phospholipid and a polyvalent cation, wherein the molar ratio of polyvalent cation to phospholipid is at least 0.05 and is sufficiently high to increase the gel-to-liquid crystal transition temperature of the particles compared to particles without the polyvalent cation.

44. the examples. A particulate composition substantially as herein described with reference to Dated this 23rd day of February 2006 ALLIANCE PHARMACEUTICAL CORPORATION By their Patent Attorneys GRIFFITH HACK Fellows Institute of Patent and Trade Mark Attorneys of Australia H; \MHrep\Keep\Spe1\F2C'(336 ;'Qc 23/02/ Th