United States Patent [191
`List et a1.
`
`USOO5389382A
`[11] Patent Number:
`[45] Date of Patent:
`
`5,389,382
`Feb. 14, 1995
`
`[54] HYDROSOLS OF PHARMACOLOGICALLY
`ACI‘IVE AGENTS AND THEIR
`PHARMACEUTICAL COMPOSITIONS
`COMPRISING THEM
`
`[75] Inventors: Martin List; Heinz Sucker, both of
`Basel, Switzerland
`
`[73] Assignee: Sandoz Ltd., Basel, Switzerland
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`4,117,118 9/1978 H'arri et a1. .
`4,540,602 10/1985 Motoyama et a1. .
`
`FOREIGN PATENT DOCUMENTS
`
`0169618 U 1985 European Pat. Off. .
`1516348 7/1978 United Kingdom .
`
`[21] Appl. No.: 642,106
`
`[22] Filed:
`
`Jan. 16, 1991
`
`[63]
`
`Related US. Application Data
`Continuation of Ser. No. 436,147, Nov. 13, 1989, aban
`cloned, which is a continuation of Ser. No. 134,337,
`Dec. 17, 1987, abandoned.
`
`Foreign Application Priority Data
`[30]
`Dec. 19, 1986 [DE] Germany ........................... .. 3643392
`
`[51] Int. GL6 ....................... .. A61K 9/14; A61K 9/ 16;
`‘
`A61K 9/50; A61K 37/02
`[52] US. Cl. .................................. .. 424/499; 424/423;
`.
`424/489; 514/11; 514/937
`[58] Field of Search ............. .. 424/422, 423, 484, 489,
`424/499; 514/11, 937
`
`OTHER PUBLICATIONS
`List et al., “Pharmaceutical Injectable Hydrosols con
`taining Water-insoluble active agents”, 63-6, Pharma
`ceuticals, 1988.
`Primary Examiner—Thurman K. Page
`Assistant Examiner-Carlos Azpuru
`Attorney, Agent, or Firm-Robert S. Honor; Melvyn M.
`Kassenoff; Carl W. Battle
`[57]
`ABSTRACI‘
`The invention provides a hydrosol of a pharmacologi
`cal active agent in an intravenous applicable, stabilised,
`pharmaceutically acceptable form, which form is sus
`pended or is dry and re-suspendable in an aqueous me
`dium.
`The hydrosol contains solid active agent particles, e. g.
`of dihydropyridines or cyclosporines.
`
`15 Claims, 1 Drawing Sheet
`
`ng /ml
`5 000-?
`
`A
`1,000 -- f,
`
`3 000 ~
`
`A
`
`C
`
`2 000"‘
`ii
`
`1000 -
`
`Actavis - IPR2017-01100, Ex. 1027, p. 1 of 7
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`

`

`US. Patent
`
`Feb. 14, 1995
`
`5,389,382
`
`F151
`
`3 000 -
`
`Actavis - IPR2017-01100, Ex. 1027, p. 2 of 7
`
`

`

`1
`
`HYDROSOLS OF PHARMACOLOGICALLY
`ACTIVE AGENTS AND THEIR
`PHARMACEUTICAL COMPOSITIONS
`COMPRISING THElVI
`
`15
`
`5,389,382
`2
`ester, e.g. Isradipine=isopropyl methyl-4-(2,l,3-ben
`zoxadiazol-4-yl)-1,4-dihydro-2,6-dimethyl-3,5-pyridine
`carboxylate or Darodipine=diethyl-4-(2,1,3-benzox
`adiazol-4-yl)
`-l,4-dihydro-2,6-dimethyl-3,S-pyridine
`dicarboxylate.
`Darodipine and Isradipine are e.g. known from the
`european patent speci?cation No. 150 and the UK pa
`tent speci?cation No. 2,037,766 respectively.
`The dihydropyridines are calcium antagonists and are
`particularly used as anti-hypertensives and for the treat
`ment of Angina pectoris.
`For their use as anti-hypertensives e. g. up to 250 mg
`preferably up to 200 mg, especially up to about 50 to 100
`mg Darodipine and up to 50 mg, preferably up to 25 mg
`e.g. 5 to 20 mg Isradipine are administered orally a day,
`e.g. 2.5 mg Isradipine twice a day. They can also be
`administered intravenously, e.g. from 0.5 to 2 mg over
`30 minutes, e. g. in the case of Isradipine.
`Other active substances in the mentioned solubility
`range include cyclosporins, especially Cyclosporin A,
`having a water solubility of below 0.004 g/ 100 ml or
`Proquazone: 1-isopropy1-7-methyl-4-phenyl-2(1H)
`quinazolone, having a water solubility of below 0.1
`g/ 100 ml.
`Further examples of compounds in the low water
`solubility range are steroids.
`The cyclosporins comprise a class of structurally
`distinct, cyclic, poly-N-methylated undecapeptides
`having valuable pharmacological, in particular immu
`nosiippressive, anti-in?ammatory and anti-parasitic, in
`particular anti-protozoal activity. The ?rst of the cy
`closporins to be isolated and the “parent” compound of
`the class, is the naturally occurring fungal metabolite
`cyclosporin, also known as cyclosporin A, the produc
`tion and properties of which are described e.g. in US.
`Pat. No. 4,117,118.
`Since the original discovery of Cyclosporin a wide
`variety of naturally occurring cyclosporins have been
`isolated and identi?ed and many further non-natural
`cyclosporins have been prepared by synthetic or semi
`synthetic means or by the application of modi?ed cul
`ture techniques. The class comprised by the cyclospo
`rins is thus now substantial and includes, for example,
`the naturally occurring cyclosporins (Thr2)-, (V all)
`and (Nva2)- Cyclosporin (also known as cyclosporins
`C, D and G respectively), as well as various semi-syn
`thetic derivatives thereof, such as their dihydro deriva
`tives (e.g. as disclosed in US. Pat. Nos. 4,108,985;
`4,210,581 and 4,220,641) including e.g. (Dihydro
`MeBmt1)-(V al2)-Cyc1osporin (also known as dihy
`drocyclosporin D) and other natural and arti?cial cy
`closporins such as those disclosed in European Patent
`Publication NO. 0,058,134 B1, for example [(D)-Ser8]
`Cyclosporin; UK Patent Application No. 2,115,936 A,
`for example [O-Acetyl-(D)-Ser8]-Cyclosporin; and En
`ropean Patent Application No. 868101122, for example
`[Val]2-[(D)Methylthio-Ser]3- and [Dihydro-MeBmtP
`[Val]2-[(D)~Methylthio-Sar]3-Cyclosp0rin.
`[In accordance with now conventional nomenclature
`for the cyclosporins, these are de?ned herein by refer
`ence to the structure of Cyclosporin (i.e. cyclosporin
`A). This is done by ?rst indicating those residues in the
`molecule which differ from those present in Cyclospo
`rin and then applying the term “Cyclosporin” to cha
`racterise the remaining residues which are identical to
`those present in Cyclosporin. Cyclosporin has the for
`mula I
`
`This is a continuation of application Ser. No.
`07/436,147, ?led Nov. 13, 1989, now abandoned, which
`in turn is a continuation of application Ser. No.
`07/134,337, ?led Dec. 17, 1987, now abandoned.
`This invention relates to hydrosols of pharmacologi
`cally active agents, suspended or re-suspendable in an
`aqueous medium.
`Hydrosols have been generally known for a long
`time. Their solid particles have diameters in the nano
`meter range, varying from about 1 nanometer (= 10"9
`meter) to about 10,000 nanometer (= 10"‘l micrometer),
`preferably to 1 micrometer.
`They can be made visible using the Tyndall effect.
`20
`The invention relates especially to a hydrosol of a
`pharmacologically, active agent in an intravenously
`applicable, stabilized pharmaceutically acceptable form,
`which form is suspended or is re-suspendable in an
`aqueous medium and is characterised in that the hydro
`sol comprises solid particles of the active substance.
`Hydrosols in an intravenously applicable pharmaceu
`tical composition form have been proposed in the UK
`Patent Speci?cation No. 1,516,348. Their particles are
`suitably modi?ed and have an appropriate diameter in
`30
`the nanometer range and aqueous suspensions thereof
`can be injected through a needle and on administration
`are taken up in the blood circulation. The suspended
`particles are sufficiently small to ?ow through blood
`vessels. They are also stabilized to avoid aggregation.
`35
`The particles described in the above-mentioned UK
`Patent Speci?cation are colloid nanoparticles. Their
`colloid material is cross-linked, e.g. cross-linked gela
`tine or a cross-linked cellulose derivative. The colloid
`particles clearly contain a water soluble or water
`insoluble pharmacologically active agent in molecular
`distribution since during their preparation, the dissolved
`active agent is not allowed to precipitate (p.5, 1.12l—p.6
`1.10).
`After administration as a suspension the pharmaco
`logically active agent is released from the nanoparticles
`at a slow rate.
`The invention provides in particular a hydrosol of
`solid active agent particles in such a form which, when
`in water suspended and administered, behaves, regard~
`50
`ing pharmacological activity, as an injectable solution,
`If pharmacologically analysed in the blood plasma,
`the pharmacological compound is detectable from a
`hydrosol of the present invention as soon as from an
`administered conventional injectable solution.
`Hitherto it was never proposed to use pharmacologi
`cally active agent particles in an aqueous hydrosol form
`for intravenous injection purposes.
`The active agent hydrosol particles of the invention
`preferably have an average statistical diameter of be
`60
`tween 1 micrometer and l nanometer, especially be
`tween 0.5 micrometer and 1 nanometer.
`Active agents for the hydrosols of the invention have
`preferably a water solubility of below 0.5 g/ 100 ml,
`especially below 0.1 g/ 100 ml at room temperature.
`Active agents in this solubility range are e.g. dihy
`dropyridines, especially those having the structure of a
`4-aryl-1,4-dihydro-2,6-dialkyl-3,S-pyridine dicarboxyl
`
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`5,389,382
`
`A-B-Sar-MeLeu-Val-MeLeu-Ala—(D)Ala—MeLeu—MeLeu—MeVal
`10
`|— l
`2
`3
`4
`5
`6
`7
`8
`9
`l l
`
`—|
`
`(I)
`
`wherein A represents the [N-methyl-(4R)-4-but-2E-en
`1-y1-4-methyl-(L)threonyl] residue of formula II
`
`CH3
`
`(II)
`
`CH2
`
`(R)
`
`no (R) cr-r
`\ /
`\
`CH3,
`CH
`
`—N-CH—CO—
`(3)
`CH3
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`which residue is abbreviated as —MeBmt— and in
`which —x—y— is —CH::CH—(trans), B is the alpha
`aminobutyric acid residue, abbreviated as —al
`phaAbu—. Accordingly (Thr2)-Cyclosporine (Cyclos
`porin C) is the compound of formula I, wherein A has
`the meaning given above and B is —Thr--, and (Dihy
`dro-MeBmt1)-(Val2)-Cyc1osporin (Dihydrocyclosporin
`D) is the compound of formula I, wherein A represents
`the -dihydro-MeBmt-residue of formula II above in
`which x—y— is —CH2-—CH2—, and B is Val-].
`As the “parent” compound of the class, Cyclosporin
`has so far received the most attention. The primary area
`of clinical investigation for Cyclosporin has been as an
`immunosuppressive agent, in particular in relation to its
`application to recipients of organ transplants, e.g. heart,
`lung, combined heart-lung, liver, kidney, pancreatic,
`bone-marrow, skin and corneal transplants and, in par
`ticular, allogenic organ transplants. In this ?eld Cyclos
`porin has achieved a remarkable success and reputation
`and is now commercially available and widely em
`ployed in clinic.
`At the same time, applicability of Cyclosporin to
`various autoimmune diseases and to in?ammatory con
`ditions, in particular in?ammatory conditions with an
`aetiology including an autoimmune component such as
`arthritis (for example rheumatoid arthritis, arthritis
`chronica progrediente and arthritis deformans) and
`rheumatic diseases, has been intensive and reports and
`results in vitro, in animal models and in clinical trials are
`wide-spread in the literature. Speci?c autoimmune dis
`eases for which Cyclosporin therapy has been proposed
`or applied include, autoimmune hematological disor
`ders (including, e.g. hemolytic anaemia, aplastic anae
`mia, pure red cell anaemia and idiopathic throm
`bocytopaenia), systemic lupus erythematosus, poly
`chondritis, sclerodoma, Wegener granulamatosis, der
`matomyositis, chronic active hepatitis, myasthenia gra
`vis, psoriasis, Steven-Johnson syndrome, idiopathic
`sprue, autoimmune in?ammatory bowel disease (includ
`ing e. g. ulcerative colitis and Crohn’s disease) endocrine
`opthalmopathy, Graves disease, sarcoidosis, multiple
`sclerosis, primary billiary cirrhosis, primary juvenile
`diabetes (diabetes mellitus type I), uveitis (anterior and
`posterior), interstitial lung ?brosis, psoriatic arthritis
`and glomerulonephritis (with and without nephrotic
`
`55
`
`65
`
`syndrome, e.g. including idiopathic nephrotic syn
`drome or minimal change nephropathy).
`A further area of investigation has been potential
`applicability as an anti-parasitic, in particular anti
`protozoal agent, with possible uses suggested including
`treatment of malaria, coccidiomycosis and schistosorni
`asis.
`Other cyclosporins exhibit the same overall pharma
`cological utility as Cyclosporin and various proposals
`for application, in particular in one or other of the
`above identi?ed indications, are prevelant in the litera
`ture, e.g. when cyclosporin is not well tolerated, e.g.
`because of hepatoxicity or nephrotoxicity in certain
`patients.
`Cyclosporin is especially useful in the ?eld of trans
`plant surgery and of autoimmune diseases where it is
`administered orally in amounts of about 50 to about 900
`mg, preferably in divided amounts, 2 to 4 times a day, of
`dosage units of 12 to 450 mg. It can also be administered
`intravenously e.g. from 225 to 375 mg per day, using a
`1:20 to 1:100 aqueous dilution of a concentration in
`ampoules which contain 1 or 5 ml of a solution which
`contains 50 mg Cyclosporin/m.
`The invention also provides a hydrosol comprising
`solid particles of a cyclosporin or of a dihydropyridine
`in a stabilized, pharmaceutically acceptable form,
`which form is suspended or is dry and re-suspendable in
`an aqueous medium.
`In order to inhibit an increase in the size of the parti
`cles of active agent in water, e.g. to prevent an increase
`in the size of the larger particles at the expense of the
`smaller particles, a stabilizer is preferably added, which
`maintains the size distribution of the active hydrosol
`particles in the dispersion constant. The stabilizer can be
`a second particulate phase.
`In the case of e. g. dihydropyridines, like Darodipine,
`Isradipine, or of Proquazone, ethyl cellulose is prefera
`bly selected as the stabilizer, although this compound
`also can be substituted partially by gelatin, which is
`another, better pharmaceutically acceptable, stabiliser
`type: Of ethyl cellulose preferably a low viscosity va
`riante is chosen, e.g. of 22 cps or e.g. 7 cps (centipoise).
`In the case of cyclosporins, e.g. Cyclosporin A, a
`gelatin is preferably selected, especially a modi?ed gela
`tin, e.g. the plasma expander GelafundinR, or a gelatin
`of a highly puri?ed collagen hydrolysate which is solu
`ble in cold water.
`In general the weight ratio of active agent to stabi
`liser is conveniently from about 1:1 to 1:50 and the
`weight ratio of active agent to water is conveniently
`from about 1:300 to 1:1500.
`For Darodipine or Isradipine the weight ratio of
`active agent:ethyl cellulose is preferably from 1:1 to 1:4,
`especially 122.5; in the case of Proquazone it is prefera
`bly from 1:3 to 1:5, especially 1:4.
`For Darodipine, Isradipine or Proquazone the weight
`ratio of active substance:water is preferably from 1:400
`to 1:600, and is in particular 1:500.
`For cyclosporins, like Cyclosporin A the weight ratio
`of active agentzgelatin is preferably from 1:5 to 1:30 and
`particularly from 1:10 to 1:30 and especially 1:20 for
`Cyclosporin A.
`
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`

`

`5
`
`15
`
`25
`
`30
`
`35
`
`5,389,382
`5
`Additionally, to the stabiliser, an acid is also prefera
`bly added as a peptisator, e.g. succinic acid or citric
`acid.
`In the case of citric acid, the weight ratio of active
`substance:citric acid is preferably from 1:8 to 1:12, and
`is especially 1:10.
`Also for cyclosporins, like Cyclosporin A the weight
`ratio of active agentzwater is preferably from 1:800 to
`121200, such as 1:1000. However, a dry hydrosol, e.g. in
`the form of a lyophilisate may also be used, especially of 10
`cyclosporins, like Cyclosporin A.
`When preparing a dry hydrosol, e.g. in the form of a
`lyophilisate, a carrier is preferably added, e. g. dextrane,
`saccharose, glycine, acetose, polyvinylpyrrolidone or
`particularly a polyol, especially mannitol. Convenient
`weight ratios of the active agent to the carrier may be
`from about 1:20 to about 1:100. In the case of mannitol,
`the weight ratio of active substancemannitol is prefera
`bly from 1:40 to 1:60, especially 1:50.
`When drying, e.g. on lyophilisation, the liquid hy
`20
`drosol,the carrier forms a network structure, which
`keeps the hydrosol particles separate and prevents their
`agglomeration.
`The invention also provides a process for the produc
`tion of a hydrosol of a pharmacologically active agent
`in an intravenously applicable, stabilised, pharmaceuti
`cally acceptable form, which form is suspended or is
`dry and resuspendable in an aqueous medium.
`Such a process is known from the above mentioned
`UK patent speci?cation No. 1,516,348. The cross-linked
`active agent containing nanoparticles, described in the
`citation, are prepared by dissolving the carrier for the
`nanoparticles, like gelatine or serum albumin, as a col
`loidal solution in water and submitting the carrier to a
`so-called desolvation process to form the nanoparticles.
`The desolvation can be effected by the addition of salts,
`e. g. Na2SO4, and/or by an alcohol, so that the resultant
`colloid particles have nanoparticle dimensions. Prefera
`bly their size is regulated by the addition of an alcohol.
`Up till this stage the active agent may be added to the
`colloid system: as an aqueous solution, if it is water
`soluble or as an organic solution, if it is dif?cultly solu
`ble in water. Preferably the active agent is dissolved
`then in an alcohol, which is used for regulating the
`desolvation stage of the colloid.
`It is believed that in the prior art process the colloid
`nanoparticles formed from the carrier bind the mole
`cules of active agent by cohaesion. No solid active
`agent particles are formed.
`The thus formed suspension is not suitable as an injec
`table solution because it contains a alcohol and/ or salt
`in an unacceptable concentration which must be
`washed out. The washing would, however, convert the
`resultant colloid particles into a colloid solution again
`because the colloid nanoparticles would destabilise.
`Such destabilisation may be avoided by cross-linking
`the carrier of the colloid nanoparticles, e. g. with an
`aldehyde, thereby ?xing their size and giving stability.
`After cross-linking and washing the system is ready
`for use and, in lyophilisated, re-suspendable state, stor
`able.
`According to the present invention active agent hy
`drosol particles may be prepared, which from a pharma
`cokinetic and a pharmacodynamic viewpoint, are as fast
`acting and behave like an injection solution.
`The hydrosol particles according to the invention are
`prepared by a process different from the prior art pro
`cess.
`
`6
`The present invention provides a process, character
`ised in that a solution in an organic solvent miscible
`with water of an active agent dif?cultly soluble in water
`is mixed with a comparatively large amount of water
`under conditions such that a colloid, insoluble in water,
`is present in the organic solvent and/or a water soluble
`colloid is present in the water thereby stabilizing the
`hydrosol of active agent to be formed, and a hydrosol of
`solid active substance particles is formed, which, if
`desired, is dried to a form, which is re-suspendable in
`water.
`One difference from the prior art process is that the
`hydrosol particles are active agent particles and not
`cross-linked gelatine or albumine particles. Also the
`hydrosol particles are bound-—when a water soluble
`colloid stabilizer is used-to exchangeable colloid mole
`cules or-when a water insoluble colloid stabilizer is
`used-to solid-but non cross-linked—colloid particles.
`Other differences are, that the desolvation process
`used in the prior art, the removal of salts and the chemi
`cal cross-linking are super?uous.
`The novel hydrosol forms may particularly be pre
`pared as follows: a solution of the active agent in a
`solvent which is miscible with water, e.g. in alcohol,
`e. g. ethanol or isopropanol, or in acetone, is mixed with
`a comparatively large amount of water, under such
`conditions that a hydrosol is produced.
`Mixing preferably is effected rapidly to promote for
`mation of the particles at the same time and in a narrow
`size distribution. A narrow size distribution is desirable
`to inhibit a re-distribution of the particles taking place in
`the suspension with the larger particles growing at the
`expense of smaller ones (=Ostwald ripening) with the
`result that the hydrosol would become more and more
`unstable. Rapid mixing also produces large numbers of
`colloidal particles.
`Permanent ?xing of the particle size however is possi
`ble if the in?uence of the organic solvent in which the
`active agent was dissolved is minimalised and for that
`reason the solvent is preferably removed.
`Removal may take place by evaporation, e.g. in a
`rotary evaporator. The hydrosol suspension, when
`practically free from organic solvent, may be used for
`intravenous injection.
`However, evaporation can also be continued so that
`the water of the hydrosol is also evaporated. Preferably
`this is effected by lyophilisation, so as to facilitate redis
`persibility.
`Upon complete evaporation of the water, a dry lyo
`philisate may be formed, especially with Cyclosporin A,
`gelatin, mannitol and acid additive. Such a lyophilisate
`is especially suitable for maintaining the stability of the
`hydrosol over a longer period.
`The lyophilisate is a starting material for the prepara
`tion of pharmaceutical compositions of different types
`and may e.g. be redispersed with distilled water, leading
`to a suspension of hydrosol particles having the same
`size distribution as of the original hydrosol. In suspen
`sion form it may be intravenously administrable. The
`dose of active agent is in general of the same order as
`used for the same active agent in conventional i.v. solu
`tions.
`The invention thus also provides the use of the hydro
`sol for the preparation of a pharmaceutical composition
`containing the hydrosol as an active agent, as well as the
`pharmaceutical compositions containing such hydro
`sols.
`
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`

`5,389,382
`7
`The invention also provides the corresponding phar
`maceutical compositions, for use in the treatment of
`diseases, e. g. of hypertension or Angina pectoris, if they
`contain the dihydropyridine hydrosols as active agents
`or for use as an immunosuppressivum or in the treat
`ment of autoimmune diseases, in?ammation conditions
`or diseases with an autoimmune component or of para
`sitic infections, using the pharmaceutical composition, if
`they contain the cyclosporin hydrosols as active agents.
`The invention also provide the method of treatment
`using the corresponding pharmaceutical compositions.
`The following examples illustrate the invention.
`
`5
`
`EXAMPLE 1
`l g of ethyl cellulose N7 (Dow Chemical) and 0.4 g of
`darodipine are dissolved in 40 ml of 94% ethanol.
`This solution is rapidly poured into 200 ml of distilled
`water of 20° C. Whilst stirring vigorously.
`Ethanol is evaporated off over 5 mins. under reduced
`pressure at 50° C. in a rotary evaporator.
`Any coarse particles which are precipitated are sepa
`rated by ?ltration (paper ?lter with a pore size of 5
`micrometers, Schleicher & Schiill).
`The average diameter of the particles is 0.116 mi
`crometers. The polydispersity factor is 28%. The mea
`surements were made on a Malvern Submicron Particle
`Analyser Type 4600 SM.
`
`25
`
`EXAMPLE 2
`30
`0.2 g of Cyclosporin A are dissolved in 8 ml 94%
`ethanol. The solution is ?ltered through a 0.2 microme
`ter membrane, and is injected through an injection nee
`dle into a vigorously stirred solution of a temperature of
`20° C. and consisting of 10.0 g of mannitol, 4.0 g of
`35
`gelatin (Stoess) of a quality which is soluble in cold
`water, and 2.0 g of citric acid in distilled water.
`Appropriate quantities are then ?lled into suitable
`containers and lyophilised.
`
`EXAMPLE 3
`0.8 g of ethyl cellullose (N22 Hercules) are dissolved
`in 29 ml of 94% ethanol, which contains 0.2 g of
`proquazone, and then poured whilst stirring into 100 ml
`of distilled water of 70° C. The ethanol is subsequently
`evaporated off for 5 mins. at 50° C., under reduced
`pressure, in a rotary evaporator.
`The average diameter of the particles is 0.152 mi
`crometers and the polydispersity factor is 2.
`The measurement was carried out using the “Nano
`sizer” from the company Coulter, which has the same
`measuring principle as the above-mentioned measuring
`device.
`In this device, the polydispersity factor is a dimen
`sionless unit of measurement for the extent of particle
`size distribution, whereby 0 is mono-disperse and 9 is
`very large variation in particle size.
`
`50
`
`55
`
`EXAMPLE 4
`60
`l g of ethyl cellulose N7 (Dow Chemical) and 0.4 g of
`progesterone are dissolved in 40 ml of 94% ethanol.
`The solution is rapidly poured into a vigorously
`stirred solution, consisting of 4.0 g of gelatin of a quality
`which is soluble in water, in 200 ml of distilled water.
`The mixture is treated further as described in Exam
`ple l. The average diameter of the particles is 0.245
`micrometer, measured as described in Example 3.
`
`65
`
`8
`EXAMPLES 5-8
`Example 1 is repeated with the difference, that 0.4 g
`of darodipine is substituted by:
`0.4 g of progesterone or
`0.4 g of dexamethasone acetate or
`0.4 g of beclomethasone dipropionate or
`0.4 g of ?uocinolon acetonide
`The average diameters of the prepared particles are:
`0.140 micrometers
`0.140 "
`0.145 "
`0.140 "
`respectively.
`The polydispersity factor is 3 for all particles.
`
`EXAMPLE 9
`l g of ethyl cellulose and 0.4 g beclomethasone dipro
`pionate are dissolved in 20 ml of 94% ethanol.
`The solution is rapidly poured into a vigorously
`stirred solution, consisting of 0.2 g of a collagen hydrol
`ysate of a quality which is soluble in cold water, in 100
`ml of water.
`The mixture is treated further as described in Exam
`ple 1. The average particle diameter is 0.12 g mikrome
`ter measured as described in Example 3.
`
`EXAMPLE l0
`1 g of ethyl cellulose N7 (Dow Chemical) and 0.4 g of
`isradipine are dissolved in 40 ml of 94% ethanol.
`The solution is rapidly poured into a vigorously
`stirred solution, consisting of 10.0 g of mannitol and 2.0
`g of citric acid in 200 ml of a plasma expander on gelatin
`basis.
`The mixture is treated further as described in Exam
`ple l. The average particle diameter is 0.320 microme
`ters measured as described in Example 3.
`Subsequently appropriate amounts of liquid are ?lled
`into suitable containers and lyophilised.
`For usage, the lyophilisates are re-dispersed with
`distilled water.
`
`EXAMPLE 11
`In tests using 5 anaesthetised rabbits, the hydrosol of
`2 mg of darodipine in 1 ml of liquid as prepared accord
`ing to example 1 was compared with a solution of 2 mg
`of darodipine in a mixture of 2 ml of ethanol and 2 ml of
`polyethylene glycol 400.
`Both samples were diluted with an isotonic glucose
`solution to a concentration of 100 micrograms per ml.
`The animals were given the samples in a quantity of
`100 micrograms of active substance per kg body
`weight, injected over 10 minutes.
`In the tests, no signi?cant systemic haemodynamic
`side-effects of the suspension on the average blood pres
`sure, pulse, blood pressure in the heart ventricle, heart
`contraction pressure, heart output volume and total
`peripheral blood circulation were detected, if compared
`with the true solution.
`
`EXAMPLE l2
`Infusions of 20 ml of a re-suspended lyophilisate,
`prepared according to Example 2 and containing 20.4
`mg of Cyclosporin A, and of 20 ml of a solution contain
`ing 30 mg Cyclosporin A and polyoxyethylated castor
`oil and alcohol as solubilisers were administered to four
`Beagle-dogs in cross-over trial design.
`
`Actavis - IPR2017-01100, Ex. 1027, p. 6 of 7
`
`

`

`After both infusions the plasma concentrations of
`cyclosporin A were recorded over 48 hours.
`For a true comparison the measured plasma values of
`the 20.4 mg Cyclosporin infusion were calculated corre
`sponding to a 30 mg infusion and shown in the graph of
`the accompanying FIG. 1, in which the plasma concen
`trations are expressed in ng/ml and the time in hours
`and in which
`Cl is the plasma concentration of the cyclosporin
`hydrosol
`A is the corresponding concentration of the cyclospo
`rin solution.
`N0 signi?cant differences in the plasma concentra
`tions of both infusion forms were found.
`The mean values of each plasma concentration series
`fell within the standard deviation ranges of the other
`series.
`We claim:
`1. A hydrosol which comprises solid particles of a
`cyclosporin and a stabilizer which maintains the size
`distribution of said particles, wherein said cyclosporin
`has a water solubility below 0.5 grams per 100 milli
`liters, and said particles have a weight ratio of cyclospo
`rin to water of about 1:300 to about 1:1500 and a weight
`ratio of cyclosporin to said stabilizer of about 1:1 to
`about 1:50.
`2. The hydrosol according to claim 1, wherein said
`cyclosporin is Cyclosporin A.
`3. The hydrosol according to claim 1, wherein the
`stabiliser is ethyl cellulose.
`4. The hydrosol according to claim 1, containing a
`pharmaceutically acceptable gelatin as the stabiliser.
`5. The hydrosol according to claim 1, in the form of
`a lyophilisate.
`
`15
`
`5,389,382
`10
`6. The hydrosol according to claim 1, containing a
`carrier wherein the weight ratio of said cyclosporin to
`said carrier is from about 1:20 to about 1:100.
`7. The hydrosol according to claim 1, with an acid
`additive.
`8. The hydrosol of claim 1 wherein said cyclosporin
`has a water solubility below 0.1 grams per 100 milliliter.
`9. The hydrosol of claim 1 wherein said solid particles
`have an average statistical diameter of between 1 mi
`crometer and 1 nanometer.
`10. A dry hydrosol in the form of solid particles com
`prising a cyclosporin and a stabilizer which maintains
`the size distribution of said particles, wherein said cy
`closporin has a water solubility below 0.5 grams per 100
`milliliters of water, and said particles have a weight
`ratio of cyclosporin to said stabilizer of about 1:1 to
`about 1:50.
`11. The hydrosol of claim 10 wherein said carrier is
`selected from the group consisting of dextran, saccha
`rose, glycine, acetose, polyvinylpyrrolidone and a
`polyol.
`12. The hydrosol of claim 1 wherein said hydrosol is
`an aqueous dispersion.
`13. A pharmaceutical composition comprising the
`hydrosol of claim 1 wherein said composition is in an
`injectable form.
`14. The hydrosol of claim 1 wherein said stabilizer is
`a second particulate phase.
`15. A pharmaceutical composition comprising solid
`particles of a cyclosporin and a stabilizer which main
`tains the size distribution of said particles, wherein said
`cyclosporin has a water solubility below 0.5 grams per
`100 milliliter, and said particles have a weight ratio of
`cyclosporin to water of about 1:300 to about 1:1500 and
`a weight ratio of cyclosporin to said stabilizer of about
`1:1 to about 1:50.
`
`25
`
`30
`
`35
`
`* * * * it
`
`45
`
`50
`
`55
`
`65
`
`Actavis - IPR2017-01100, Ex. 1027, p. 7 of 7
`
`