United States Patent [19]
`Kee et al.
`
`US005369095A
`Patent Number:
`Date of Patent:
`
`' [111
`
`[451
`
`5,369,095
`Nov. 29, 1994
`
`[54]
`
`[75]
`
`[73]
`
`[21]
`[22]
`
`[63]
`
`[51]
`
`[52]
`[58]
`
`[56]
`
`COMPOSITIONS AND METHOD
`COMPRISING SUBSTITUTED GLYCOSIDES
`AS MUCUS MEMBRANE PERMEATION
`ENHANCERS
`Inventors: Tai-Lee Kee, Grand Prairie, Tex.;
`Jack M. Shaw, Mechanicsville, Va.
`Assignee: Alcon Laboratories, Inc., Fort
`Worth, Tex.
`Appl. No.: 31,000
`Filed:
`Mar. 12, 1993
`
`Related US. Application Data
`Continuation of Ser. No. 745,136, Aug. 13, 1991, aban
`doned, which is a continuation-in-part of Ser. No.
`480,471, Feb. 14, 1990, abandoned.
`
`Int. Cl.5 ................... .. A61K 9/10; A61K 31/715;
`A61K 31/725; A61K 31/79
`. 514/24
`US. Cl.
`Field of Search ................. .. 514/52, 57, 317, 392,
`514/402, 915, 960, 946, 947, 777, 781, 772, 780,
`779, 772.3, 24, 25, 42, 53; 536/41, 23, 53, 54
`References Cited
`U.S. PATENT DOCUMENTS
`
`4,215,130 7/ 1980 Sjoerdsma ......................... .. 514/392
`4,237,141 12/ 1980 Shiozawa et a1.
`514/317
`
`4,558,063 12/1985 Beeley ct a1. . . . . . . .
`
`. . . . . . .. 514/402
`
`252/ 174.17
`4,722,837 2/ 1988 Cameron
`...... .. 514/ 166
`4,725,590 2/ 1988 Ritchey
`4,742,083 5/ 1988 Ritchey ..................... ..
`514/621
`4,921,838 5/ 1990 Catsimpoolas et al.
`.... .. 514/25
`4,923,693 5/ 1990 Michalos ................... ..
`514/915
`4,942,038 7/ 1990 Wallach .... ..
`424/450
`
`5,041,450 8/1991 Chiou et a1. . . . . . . . .
`
`. . . . . . .. 514/915
`
`252/174.17
`5,062,989 11/1991 Kamegai et a1.
`5,219,887 6/1993 Andrews et a1. ................. .. 514/881
`
`5,262,178 ll/1993 Malfroy Camine et al. ..... .. 435/212
`
`FOREIGN PATENT DOCUMENTS
`
`1-151528 6/1989 Japan .
`
`OTHER PUBLICATIONS
`Tonjum, Asbjorn M., “Permeability of Rabbit Corneal
`Epithelium in Horseradish Peroxidase After the In?u
`ence of Benzalkonium Chloride,” Acta Ophthal
`mologica, v01. 53, p. 335 (1975).
`Morimoto et al., “Evaluation of permeability enhance
`ment of hydrophilic compounds and macromolecular
`compounds by bile salts through rabbit corneas in vi
`tro,” J. Pharm. PharmacoL, v01. 39, p. 124 (1987).
`Schulte, Thomas L., “Ophthalmic compositions con
`taining biphenamine and their use in the treatment or
`prevention of in?ammation,” Chemical Abstracts, vol.
`106, 125931t, p. 402 (1987).
`Grass et al., “Mechanisms of Corneal Drug Penetration
`1. In Vivo and In Vitro Kinetics,” Journal of Pharma
`ceutical Sciences, vol. 77, No. 1, p. 3 (Jan, 1988).
`Newton et al., “Topically Applied Cyclosporine in
`Azone Prolongs Corneal Allograft Survival,” Investiga
`tive Ophthalmology & Visual Science, vol. 29, No. 2, p.
`208 (Feb., 1988).
`Primary Examiner-Ronald W. Grif?n
`Attorney, Agent, or Firm—Sally Yeager
`[57]
`ABSTRACT
`The use of substituted glycosides to enhance the pene
`tration of drugs across mucus covered epithelial tissues
`of humans and animals is disclosed, including enhanced
`penetrations of topically applied ophthalmic drugs
`through the corneal -' epithelium of said humans and
`animals.
`
`10 Claims, 2 Drawing Sheets
`
`AQUESTIVE EXHIBIT 1016 page 0001
`
`

`

`US. Patent
`
`Nov. 29, 1994
`
`Sheet 1 of 2
`
`5,369,095
`
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`
`AQUESTIVE EXHIBIT 1016 page 0002
`
`

`

`US. Patent
`
`Nov. 29, 1994
`
`Sheet 2 of 2
`
`5,369,095
`
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`
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`
`AQUESTIVE EXHIBIT 1016 page 0003
`
`

`

`1
`
`COMPOSITIONS AND METHOD COMPRISING
`SUBSTITUTED GLYCOSIDES AS MUCUS
`MEMBRANE PERMEATION ENHANCERS
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`This is a continuation of US. Ser. No. 07/745,136,
`?led Aug. 13, 1991 (now abandoned), which is a con
`tinuation-in-part of US. Ser. No. 07/480,471, ?led Feb.
`14, 1990 (now abandoned).
`FIELD OF THE INVENTION
`This invention relates to substituted glycosides as
`enhancers for the permeation of therapeutic agents
`through mucus membranes of humans and animals.
`
`35
`
`40
`
`BACKGROUND OF THE INVENTION
`The present invention relates to the ?eld of drug
`delivery across mucus covered epithelial tissues of hu
`mans and animals. Such tissues include nasal, pulmo
`nary, rectal, buccal, vaginal, uteral, and gastrointestinal
`routes for drug administration. More particularly, this
`invention relates to enhancement of the penetration of
`ophthalmic drugs and other therapeutic agents through
`25
`the cornea and other tissues of the eye such as the sclera
`and conjunctiva of humans and animals.
`In order for an ophthalmic drug to be therapeutically
`effective, it is generally necessary for the drug to pene
`trate the cornea and be taken up in the aqueous humor,
`ciliary processes and other tissues in the eye. There are
`notable exceptions to this general rule, such as drugs or
`drug products which produce a localized therapeutic
`effect by acting on the exterior surface of the cornea
`(e. g., drugs or drug products useful in improving ocular
`comfort and/or treating dry or irritated eyes). How
`ever, the treatment of conditions involving physiolog
`ical mechanisms within the eye (e. g., glaucoma, diabetic
`retinopathy, cataracts, etc.) generally does require the
`permeation of topically applied ophthalmic drugs pri
`marily through the cornea.
`In order for a drug to pass through the cornea, it must
`penetrate three layers of tissue, namely, the epithelium,
`stroma, and the endothelium. Except for highly lipo
`philic drugs, the epithelium is the main barrier to drug
`45
`penetration of the cornea. Penetration of the stroma
`basically involves diffusion of the drug through a bar
`rier which is approximately 360 microns thick. There
`are currently no known methods of enhancing drug
`penetration through the stroma or endothelium. How
`ever, it is possible to enhance the penetration of drugs
`through the epithelium, and thereby enhance the over
`all absorption of drugs applied topically to the eye. The
`present invention is directed to such enhancement.
`There have been prior attempts to enhance the pene
`55
`tration of drugs through the corneal epithelium. The
`goal of such attempts has generally been to enhance
`penetration of drugs through the corneal epithelium to
`an optimal point at which the stroma alone controls
`drug transport through the corneas. The prior attempts
`have included use of chemical agents to enhance the
`penetration of drugs through the epithelium. It has been
`reported that benzalkonium chloride (BAC), bile salts,
`dimethyl sulfoxide (DMSO), ethylenediamine tetraace
`tate (EDTA) and l-dodecylazayl-cycloheptan-2-one
`(AZONE ®) enhance the corneal penetration of cer
`tain drugs. The following publications may be referred
`to for further background concerning the use of such
`
`50
`
`60
`
`5,369,095
`2
`agents to enhance corneal penetration: Acta Ophthal
`mologica, Vol. 53, p.335 (1975); J. Pharm. PharmacoL,
`Vol.39, p.124 (1987); Chem. Abstracts, Vol.106, 12593 lt,
`p.402 (1987); Journal of Pharmaceutical Sciences, Vol.77,
`No.1 (Jan.,l988); and Investigative Ophthalmolog and
`Visual Science, Vol.29, No.2 (Feb.,1988). Notwithstand
`ing such prior attempts, there continues to be a need for
`a means of safely and effectively enhancing the penetra
`tion of drugs through the cornea.
`
`SUMMARY OF THE INVENTION
`A principal objective of the present invention is to
`provide for a method of enhancing drug delivery across
`mucus covered epithelial tissues, particularly those of
`the cornea, sclera and conjunctiva of humans and ani
`mals. A further objective of the present invention is to
`provide topical ophthalmic compositions containing
`one or more agents for enhancing the penetration of the
`active ingredient(s) contained therein.
`The foregoing objectives and other general objec
`tives of the present invention are satis?ed by the provi
`sion of a means of enhancing penetration by using a
`class of compounds collectively referred to herein as
`substituted glycosides to enhance the penetration of
`ophthalmic drugs through the corneal epithelium,
`sclera and conjunctiva. In addition, the objectives of the
`present invention are furthered when viscosity enhanc
`ing polymers are used in conjunction with the substi
`tuted glycosides and ophthalmic drugs so that the com
`positions are retained in the eye for a relatively longer
`period of time, thus allowing the enhancers more time
`to facilitate drug transport through the cornea, sclera
`and conjunctiva.
`
`BRIEF DESCRIPTION OF THE FIGURES
`FIG. I compares the amount of a drug, para-amino
`clonidine, found in the aqueous humor of rabbits which
`were administered, the drug with and without the sub
`stituted glycoside enhancer, dodecyl maltoside.
`FIG. II compares the amount of a drug, para-amino
`clonidine, found in the aqueous humor of rabbits which
`were administered the drug as a solution, or the drug in
`combination with a viscosity enhancing polymers, hy
`droxypropylmethyl cellulose (HPMC) and dodecyl
`maltoside, or the drug with HPMC.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`The present invention is based on the discovery that
`substituted glycosides effectively and safely enhance
`the corneal penetration of ophthalmic drugs. When
`“corneal penetration” is used herein it includes penetra
`tion through the cornea, sclera and conjunctiva of the
`eye. These penetration enhancers can be used in compo
`sitions comprising any ophthalmic drug which, to be
`effective, must be substantially taken up by the aqueous
`humor, ciliary processes and other tissues in the eye
`upon topical administration. Examples of classes of
`ophthalmic drugs with which the substituted glycosides
`of the present invention can be used, include: steroids,
`growth factors, cycloplegics, miotics, mydriatics, thera
`peutic proteins and peptides, antioxidants, aldose reduc
`tase inhibitors, nonsteroidal antiinflammatories, im
`munomodulators, antiallergics, antimicrobials, angio
`static agents and anti-glaucoma therapeutic agents.
`
`AQUESTIVE EXHIBIT 1016 page 0004
`
`

`

`5,369,095
`3
`The penetration enhancing substituted glycosides
`used in the present invention have the following struc
`ture:
`
`4
`-continued
`
`cH2oH
`
`HO
`
`H OH
`
`CHZOH
`
`I
`
`HO
`
`H OH
`
`CHZOH
`
`E
`
`on H STR‘
`HO
`
`HOH
`
`[B]
`
`[c]
`
`[D]
`
`15
`
`20
`
`25
`
`wherein R1 is a hydrophobic group including saturated
`and unsaturated aliphatic hydrocarbon groups which
`range from 8 to 28 carbons in length with 1 to 5 double
`bonds. The aliphatic hydrocarbon group can be a
`straight or branched chain and may be substituted by
`one or more aromatic, cycloaliphatic or hydrophilic
`(e.g. hydroxyl, thiol, ester or amino) groups. R; is a
`group derived from any cyclic or acyclic saccharide
`containing 4-7 carbons and their isomers;
`X is an integer from 1-10; and
`Z is an oxy (—O—), carbonyloxy
`
`phosphoryl
`
`thio (--S—), or
`carboxamido
`
`-——oH
`CHZOH
`
`35
`
`CHIOH
`CHZOH
`H OH H
`0
`
`where R; covalently bound to such group.
`More speci?cally R1 can be a straight 8-18 carbon
`alkyl chain in hemiacetal linkage (glycoside) to the
`saccharide; and R; a group derived from any of a vari
`ety of isomeric saccharides containing 5 or 6 carbons.
`The saccharide can be, for example, an aldehyde-con
`taining saccharide (glucose, mannose, arabinose, galac
`tose, xylose); a ketone-containing saccharide (fructose,
`xylulose, sorbose); a saccharide alcohol (sorbitol, inosi~
`tol, xylitol, mannitol); a saccharide acid (glucuronic
`acid, neuramic acid, mannuronic acid); a deoxysaccha
`ride (deoxy-ribose, rhamnose,); an aminosaccharide
`(glucosamine, galactosamine). Higher order saccharides
`being covalently linked in any of a number of ways to
`form different isomeric structures include for example
`disaccharides such as maltose, cellobiose, sucrose and
`lactose and trisaccharides, such as raffmose.
`The preferred penetration enhancers are alkyl chain
`containing glycosidas derived from maltose and glucose
`with R1 being 8 to 18 carbons and having the following
`structures:
`
`55
`
`CHZOH
`
`O
`
`H
`H
`OH H
`HO
`
`o
`O //
`—\
`R1
`
`H 0H
`
`[A]
`
`65
`
`H OH
`
`H OH
`
`The most preferred penetration enhancer is:
`
`CHZOH
`
`cH2oH
`
`Dodecyl Maltoside
`
`H
`O H H
`0
`EH H
`gH H 0-K‘
`Ho
`
`0
`
`H on
`
`H OH
`
`The substituted glycosides which are useful in the
`present invention may be described as being “am
`phipathic”, since they include both hydrophilic and
`hydrophobic groups. While not wishing to be bound by
`any theory, it is believed that substituted glycosides
`enhance the corneal penetration of drugs by partition
`and interaction with protein, glycoprotein and lipid
`components present in the membrane of the corneal
`epithelium. Such interaction is believed to alter the
`degree of order of the proteins and lipids in the mem
`brane, thereby modifying the function of the epithelium
`as a barrier to drug penetration. Whatever the mecha
`nism, the net result is that drug penetration across the
`epithelium is enhanced.
`The use of substituted glycosides in accordance with
`the present invention to enhance corneal penetration of
`drugs signi?cantly increases the amount of drug which
`
`AQUESTIVE EXHIBIT 1016 page 0005
`
`

`

`5,369,095
`5
`6
`hydroxypropylmethylcellulose (HPMC), methylcellu
`is able to penetrate the cornea. The degree of enhance
`lose, polyvinyl alcohol (PVA), polyvinyl pyrrolidone,
`ment will vary with different drugs, but in some cases
`carboxymethylcellulose and agarose. In addition, prote
`may be as much as 3-fold or mere. Because drugs can
`more effectively penetrate the cornea, less drug is lost
`ins, synthetic polypeptides and polymer-peptide co
`due to ?ow down the punctum and therefore less drug
`polymers which enhance viscosity and are ophthalmi
`need be administered to effectively treat a particular
`cally acceptable can be used to increase the viscosity of
`indication. This is particularly bene?cial when it is nec
`the compositions to provide for better bioavailability.
`essary to administer drugs which cause severe systemic
`Typically, proteins which can be used include: gelatin,
`side effects.
`collagen, albumin and casein.
`The amount of substituted glycoside required in
`The preferred viscosity enhancing agents are one or
`order to enhance cornea penetration will depend on
`more polymers selected from: PVA, HPMC and HEC.
`various factors, such as the solubility, partition coef?ci
`The most preferred agent is HPMC. The viscosity en
`ent and molecular weight of the ophthalmic drug or
`hancing agents are added to provide for compositions
`therapeutic agent; the excipients (surfactants, preserva
`with a viscosity of between about 50 and 300 cps.
`tives, viscosity enhancing polymers) present in the for
`The preferred method for enhancing the penetration
`mulation; and the particular enhancer being used. In
`of a drug or therapeutic agent comprises the use of
`general, the more lipophilic the drug to be delivered,
`dodecyl maltoside at a concentration of about 0.01% to
`the less enhancer is required to increase penetration,
`1.0% in combination with the polymer, HPMC, in an
`and the higher the concentration of the substituted gly
`amount sufficient to provide a composition with a vis
`coside, the’ better the corneal penetration. Typically,
`cosity of about 50 to about 300 cps.
`one or more enhancers will be used in an amount of
`The following examples further illustrate the compo
`from about 0.01% to about 20% (weight/volume) pref
`sitions which, according to the present invention, com
`erably from about 0.01 to 1.0%.
`prise the corneal penetration enhancing properties of
`The substituted glycosides can be used with certain
`the substituted glycosides and their use to enhance cor
`topical drug delivery systems wherein an excipient or
`nea penetration.
`vehicle will not substantially impair or prevent the sub
`stituted glycosides from functioning as corneal penetra
`tion enhancers. For example, the substituted glycosides
`can be formulated in compositions which are solutions,
`suspensions, ointments, gels or ?lms. The, type of com
`position will depend on, among other things, the chemi
`cal and physical properties of the drug or therapeutic
`agent to be delivered and the properties of polymeric
`materials used in the formulation. These properties are
`well known to a person of ordinary skill in the art of
`drug formulation and delivery.
`In a preferred embodiment, the present invention
`further comprises the use of viscosity enhancing poly
`mers in conjunction with the substituted glycosides to
`enhance ocular bioavailability. The longer a topical
`ophthalmic formulation is in contact with the eye the
`better the ocular bioavailability. Through the use of
`polymers in conjunction with the above described en
`hancers the compositions of the present invention are
`retained on the cornea longer. As a result, the penetra
`45
`tion enhancing components of the compositions can
`more effectively interact with the corneal epithelium to
`enhance penetration of the desired drugs or therapeutic
`agents into the eye. It has been found that the use of
`polymers in conjunction with substituted glycosides can
`provide for up to about a 3 to 10 fold increase in the
`amount of drug or therapeutic agent made available to
`the tissues. The effectiveness of the therapeutic agent
`and the substituted glycosides can be improved when
`the viscosity of the compositions is increased up to
`about 1000 centipoise (cps), preferably between about
`50 cps. to 300 cps. Polymers are added to provide for
`this desired viscosity increase.
`Any synthetic or natural polymer which will increase
`viscosity and is compatible with tissues of the eye and
`the ingredients of the substituted glycoside composi
`tions can be used. Such polymers are referred to herein
`as “viscosity enhancing, ophthalmically acceptable pol
`ymers.” Examples include, but are not limited to: natu
`ral polysaccharides and gums, such as alginates, carra
`geenans, guar, karaya, locust bean, tragacanth and xan
`than; and synthetic polymers, such as carbomer, hy
`droxyethylcellulose (HEC), hydroxypropylcellulose,
`
`Procedure for Preparation of Formulation
`Approximately 85% (8.5 ml) of the batch volume of
`puri?ed water was added to a container. All of the
`ingredients were added to the container: 0.018 g mono
`basic sodium phosphate; 0.012 g dibasic sodium phos
`phate; 0.33 g mannitol; 0.1 ml of 1.0% BAC; 0.001 g
`disodium edetate; 0.0125 g para-aminoclonidine. The
`ingredients were mixed well and stirred until all ingredi
`ents dissolved into a solution. 0.005 g dodecyl maltoside
`was added to the container and sonicated for 5 minutes.
`The pH was adjusted to pH 6.5. Puri?ed water was then
`poured through a sterilizing ?lter into the container
`(q.s. to 10 ml) and the solution was mixed well.
`Nine New Zealand albino rabbits were selected for
`evaluation of the penetration through the cornea of the
`para-amino-clonidine formulation set forth above. All
`rabbits received 30 ul of the 0.125% para-amino-cloni
`dine topically in both eyes. Three rabbits were sacri
`?ced at 20 minutes from dosing and aqueous humor was
`withdrawn from their eyes. The aqueous humor as as
`sayed by liquid scintillation counting to determine the
`amount of para-amino-clonidine in the aqueous humor.
`The same procedure was done on 3 different rabbits at
`60 minutes from dosing and on another 3 rabbits, 120
`minutes from closing. Nine control rabbits received
`
`25
`
`60
`
`65
`
`EXAMPLE 1
`The following formulation is an example of a topical
`ophthalmic composition which can be used to treat
`glaucoma.
`
`_F_<>_r21.l11a_ti2a_
`
`Ingredients
`Para-amino-clonidine
`Dodecyl maltoside
`Benzalkonium chloride
`Disodium Edetate, USP
`Sodium phosphate, monobasic, USP
`Sodium phosphate, dibasic, USP
`Mannitol, USP
`HCI, NF and/or NaOH, NF
`Puri?ed Water, USP
`
`% (weight/volume)
`0.125
`0.050
`0.01
`0.01
`0.18
`0.12
`3.3
`q.s. pH to 6.5 i 0.2
`q.s. 100
`
`AQUESTIVE EXHIBIT 1016 page 0006
`
`

`

`5,369,095
`8
`7
`junction with HPMC or with HPMC and dodecyl mal
`0.125% para-aminoclonidine as set forth in the formula
`tion above without 0.05% dodecyl maltoside. Aqueous
`toside, respectively, as compared to those which re
`humor was withdrawn and assayed as explained above.
`ceived the drug without HPMC or HPMC and dodecyl
`The results are shown in the graph depicted in FIG. I.
`maltoside. The results indicate that dodecyl maltoside
`It can be seen from the graph that the amount of para
`enhances the penetration of para-amino-clonidine
`amino-clonidine in the aqueous humor is greater in the
`through the cornea over HPMC alone and para-amino
`rabbits treated with the formulation containing dodecyl
`clonidine alone.
`maltoside. At 60 minutes there is almost a four fold
`increase in the amount of para-aminoclonidine found in
`the aqueous humor of those rabbits which received the
`drug in conjunction with dodecyl maltoside versus
`those who received the drug without dodecyl malto
`side. Therefore, the results indicate that dodecyl malto
`side enhanced penetration of para-aminoclonidine
`through the cornea.
`
`15
`
`EXAMPLE 2
`
`M
`% (Weight/volume)
`0.125
`3. l
`
`Ingredients
`Para-amino-clonidine
`Hydroxypropylmethylcellulose-ESOLV,
`(HPMC) USP
`Dodecyl maltoside
`Benzalkonium chloride
`Disodium Edetate, USP
`Sodium phosphate, monobasic, USP
`Sodium phosphate, dibasic, USP
`Mannitol, USP
`RC1, NF and/or NaOH, NF
`Puri?ed Water, USP
`
`0.05
`0.01
`0.01
`0.18
`0.12
`3.3
`q.s. pH to 6.75 i 0.2
`q.s. 100
`
`25
`
`30
`
`45
`
`Procedure for Preparation of Formulation
`Approximately 85% (8.5 ml) of the batch volume of
`puri?ed water was added to a container. All of the
`ingredients were then added to the container: 0.018 g
`35
`monobasic sodium phosphate; 0.33 g mannitol; 0.1 ml of
`1.01% BAC; 0.001 g disodium edetate; 0.0125 g para
`amino—clonidine; 0.31 g HPMC. The ingredients were
`mixed well. 0.005 g dodecyl maltoside was added to the
`container and sonicated 5 minutes. The pH was adjusted
`to pH 6.5. Puri?ed water was then poured through a
`sterilizing ?lter into the container (q.s. to 10 ml) and the
`solution was mixed well.
`Nine New Zealand albino rabbits were selected for
`evaluation of the penetration through the cornea of the
`para-amino-clonidine formulation set forth above. All
`rabbits received 30 ul of the 0.125% para-amino-cloni
`dine topically in both eyes. Three rabbits were sacri
`?ced at 20 minutes from dosing and their aqueous
`humor was withdrawn from their eyes. The aqueous
`humor was assayed by liquid scintillation counting to
`determine the amount of para-amino-clonidine in the
`aqueous humor. The same procedure was done on three
`different rabbits at 60 minutes from dosing and on an
`other three rabbits, 120 minutes from dosing. Nine con
`trol rabbits received 0.125% para-amino-clonidine as set
`forth in the formulation above without 0.05% dodecyl
`maltoside and 3.1% HPMC. Another 9 rabbits received
`0.125% para-aminoclonidine as set forth in the formula
`tion above without 0.05% dodecyl maltoside. Aqueous
`humor was withdrawn and assayed as explained above.
`The results are shown in the graph depicted in FIG. II.
`It can be seen from the graph that the amount of para
`amino-clonidine in the aqueous humor is greater in the
`rabbits treated with the formulation containing HPMC
`and HPMC with dodecyl maltoside. At 60 minutes
`there is almost a 4.0 fold and a 10 fold increase in the
`amount of para-amino-clonidine found in the aqueous
`humor of those rabbits which received the drug in con
`
`50
`
`55
`
`EXAMPLE 3
`The following carbachol formulations were prepared
`and evaluated for Acute Pupil Diameter Response in
`New Zealand Albino Rabbits. Animals were restrained
`for the duration of a study. For 30 minutes animals were
`adapted to room lighting and then the eyelashes were
`trimmed. Two baseline pupil diameter measurements
`were taken with a hand-held micrometer on one eye of
`each animal. This eye was then closed with one 30 mi
`croliter aliquot of each formulation and subsequent
`pupil diameter measurements taken at 0.5, l, 2, 3, 4, and
`5 hours. The results are shown in Table 1.
`
`Formulation
`Compound
`
`NaI-I2PO4
`Na2HPO4
`EDTA
`BAC
`Carbachol
`Dodecyl Maltoside
`HPMC
`Water
`pH was adjusted to 6.5
`
`A
`wt. %
`
`0.18
`0.12
`0.01
`0.01
`0.3
`0
`0
`q.s. to 100
`
`B
`wt. %
`
`C
`wt. %
`
`0.18
`0.12
`0.01
`0.01
`0.3
`0.07
`3.3
`q.s. to 100
`
`0.18
`0.12
`0.01
`0.01
`3
`0
`0
`q.s. to 100
`
`TABLE I
`MEAN PERCENT CHANGE IN RABBIT PUPIL
`DIAMETER
`Time after treatment (hrs)
`1
`2
`3
`4
`
`0
`
`Q
`
`Formulation
`
`5
`
`—1
`O —22 —28 —20 --13 —5
`A (0.3% Carbachol)
`B (0.3% Carbachol + 0 —63 —66 _--61 —56 —36 —23
`0.07 DDM + 3.3%
`HPMC)
`C (3% Carbachol)
`
`0 —45 —44 —35 —27 —15 - 10
`
`As evident from the data, Formulation B containing
`Carbachol with dodecyl maltoside and hydroxypropyl
`methylcellulose is far superior to control (Formulation
`A) and to a formulation (Formulation C) with 10 times
`higher drug concentration. Formulation B is useful for
`the treatment of glaucoma.
`
`EXAMPLE 4
`The procedure in Example 3 was repeated, except
`Carbachol was substituted with 1% amount by weight
`of Pilocarpine in Formulations A and B and with 4%
`Pilocarpine in Formulation C. The results are shown in
`Table 2.
`
`TABLE 2
`MEAN PERCENT CHANGE IN RABBIT PUPIL
`DIAMETER
`Time after treatment (hrs)
`1
`2
`3
`4
`
`0
`
`Q
`
`Formulation
`
`0
`0 —37 —26 —15 —5
`A(1% Pilocarpine)
`B (1% Pilocarpine + 0 —45 —38 —28 —17 —6
`0.07% DDM + 3.3%
`HPMC)
`
`5
`
`0
`0
`
`AQUESTIVE EXHIBIT 1016 page 0007
`
`

`

`5,369,095
`
`10
`
`9
`TABLE 2-continued
`MEAN PERCENT CHANGE IN RABBIT PUPIL
`DIAMETER
`Time after treatment (hrs-l
`l
`2
`3
`4
`
`0
`
`i
`
`Formulation
`.
`.
`C (4% Pllocarplne)
`
`5
`
`S
`
`0 —37 ~34 —23 —8 —5 —l
`
`The results clearly indicate that Pilocarpine Formula-
`tion B with the enhancer dodecyl maltoside and a vis- l0
`cosity enhancing agent, HPMC, shows superior perme-
`ation to control formulations. Formulation B is useful in
`the treatment of glaucoma.
`
`EXAMPLE 5
`The procedure of Example 3 was repeated, except
`Carbachol was substituted with 1% amount by weight
`of Epinephrine in Formulation A and 0.3 and 3% in
`Formulations B and C respectively. The results are
`shown in Table 3.
`
`15
`
`20
`
`Formulation
`A (wt. %)
`Na Acetate Trihydrate 0.03
`Acetic Acid 6N
`0.0043 to pH 4.5
`Mannitol
`4.6
`BAC
`Q01
`EDTA
`0.05
`gfggg?xizgmside
`83g
`HPMC
`0,00
`Water
`q-S- to 100
`
`Formulation
`(B (wt. %)
`0.03
`0.0043 to pH 4.5
`4.6
`0'01
`0.05
`3:39,
`330
`q-S- to 100
`
`TABLE 5
`Aqueous Humor Concentration of Cipro?oxacin
`(FE/m1)
`_
`30 mm"
`60 mm‘
`13-99
`24-10
`108'78
`25077
`
`_
`120 mm‘
`34-37
`39495
`
`FormulationA
`Fmmulam“ B
`
`TABLE 3
`B T
`E IN
`ME PE
`AN RCENTE?IALMEUEER RAB I PUPH‘
`Time after treatment (hrs)
`%
`1
`2
`3
`4'
`
`o
`
`Formulation
`_
`0
`A (1% Epinephrine)
`B(O.3% Epinephrine + 0
`
`0
`53
`
`0
`63
`
`0
`60
`
`0
`5o
`
`0
`40
`
`It can be seen that Formulation B containing Cipro
`floxacin, the enhancer dodecyl maltoside and viscosity
`enhancing agent HPMC is 10 times more permeable
`through the cornea as compared to control formulation
`25 A. Formulation B is useful in the treatment of bacterial
`conjunctivitis and corneal ulcers.
`
`EXAMPLE 3
`
`5
`
`o
`2s
`
`0.07% DDM +
`
`_
`
`_
`
`_
`
`_
`
`_
`
`_
`
`3.3% HPMC)
`C (3% Epinephrine)
`
`0
`
`22
`
`22
`
`15
`
`1°
`
`1°
`
`6
`
`45
`
`DIAMETER
`
`Formulation
`A (0.25% Phenylephrine)
`B (0.25 Phenylephrine +
`0.07% DDM + 3.3% HPMC)
`c (2.5% Phenylephrine)
`
`o
`
`30 36 26 2o 10
`
`7
`
`The results show that Formulation B, useful in the
`treatment of glaucoma, is far superior even at l/ 10th
`concentration when formulated with enhancer dodecyl
`maltoside and the viscosity enhancing agent, HPMC,
`and shows higher permeation through corneal epithe- 6O
`lium than control.
`
`EXAMPLE 7
`The following Cipro?oxacin formulations were pre-
`pared and their permeation through rabbit cornea was 65
`evaluated by measuring the concentration in aqueous
`humor according to the procedure described in Exam~
`ple 2. The results are shown in Table 5.
`
`The results clearly indicate that the formulations
`containing 0.01 and 0.1% of dodecyl maltoside showed
`superior permeation (7.5 and 22.1) fold respectively) for
`Atenolol as compared to control.
`
`EXAMPLE 9
`The procedure in Example 8 was repeated with the
`following para-amino clonidine formulations.
`
`AQUESTIVE EXHIBIT 1016 page 0008
`
`Albino rabbits were sacrificed, and wlthm 15 minutes,
`30 the corneas were mounted and clamped between diffu
`sion cells according to the published procedure by
`sch9enwald et a1- Q' pl'farm- S91- 72’ 126_6’_ 1983)- 7 ml
`From the data it is evident that Formulation B with
`of blcaFbonated Rmger s sohmon ‘:qnmng reduced
`the enhancer dodecyl maltoside and viscosity enhanc-
`glutathione was added to the endothelial side to serve as
`ing agent HPMC permeates far better through the cop
`nea than Formulation c with 10 fold higher drug con- 35 the twelve‘ S_°!u“°n- An equal v°lume °_f the Same
`centration. Formulation B is useful in the treatment of
`Solutlon contammg‘Atenolol’ a ?'blocker’ wlth or ‘lath’
`glaucoma.
`out dodecyl maltoside (DDM) was added to the ep1the
`lial side to serve as the donor solution. The apparent
`permeability coef?cient was calculated from the aver
`EXAMPLE 6
`The procedure of Example 3 was repeated’ except 4'0 age cumulative amount of drug penetrating through the
`Carbachol was substituted with 0.25% amount by
`°°mlfafm1fr Fme accofdmfg t° slchPenwald
`weight of Phenylephrine in Formulations A and B and
`T e o Owmg sohmon 0mm anons were used‘
`2.5% in Formulation C. The results are shown in Table
`4-
`
`TABLE 4
`Eff‘
`MEAN PERCENT CHANGE IN RABBIT PUPIL
`CaClzlHzO
`_
`mm; MgCl2.6l-l2O
`0
`g
`1
`2
`3
`4
`5
`50 NaH2PO4
`o
`5
`9
`o
`0
`0
`o
`gj‘Hco3
`hi
`0
`s3 55 41 25
`9
`3
`R ‘awed G1
`Ac “f1 “m °“e
`Die” °
`.
`odecyl Maltoslde
`55 Water q.s.
`
`'
`
`'
`
`B
`A (Control)
`(wt. %)
`(wt. %)
`383;,
`832%
`0.0153
`0.0153
`0.0159
`0.0159
`0.0103
`0.0103
`8'33;
`333(5);
`60092
`0'0092
`0'03
`0'03
`'
`'
`0.01
`0
`100
`100
`Permeability coefficient (cm/sec x 10-6)
`Formulation A
`2.3
`B
`17-3
`c
`50'8
`
`'
`
`c
`(wt. %)
`38229
`0.0153
`0.0159
`0.0103
`8'33:
`0'00”
`0'03
`'
`0.10
`100
`
`

`

`11
`
`Formulation
`05% para-amino clonidine
`05% para-amino clonidine + 0.01% DDM
`05% para-amino clonidine + 0.1% DDM
`
`permeability
`coefficient
`(cm/sec X 10316)
`3.8
`22.0
`39.0
`
`5,369,095
`12
`1. A method of enhancing the penetration of a drug
`across mucus covered epithelial tissues, which com
`prises:
`topically applying to the mucus covered epithelial
`tissue a pharmaceutical composition comprising a
`therapeutically effective amount of the drug and an
`amount of a substituted glycoside effective to en
`hance penetration of the drug across the mucus
`covered epithelial tissue said substituted glycoside
`having the formula:
`
`The results clearly indicate that the formulations
`containing dodecyl maltoside showed superior perme
`ation for para-amino clonidine as compared to control.
`
`EXAMPLE 10
`The procedure of Example 8 was repeated with
`Atenolol except dodecyl maltoside was substituted with
`0.1% amount by weight of each of the following substi
`tuted glycosides of this invention and the following
`results were obtained.
`
`15
`
`Enhancer
`'I‘etradecyl maltoside
`Decyl maltoside
`Nonyl B-D-glucoside
`Decyl ?-D-glucoside
`Undecyl B-D-glucoside
`Dodecyl B-D-glucoside
`Octyl thioglucoside
`Decanoyl N-methylglucosamide
`Sucrose monolaurate
`Lysophosphatidyl Inositol, 0.01%
`
`Permeability Coefficient
`(cm/sec X l0_6)
`16.8
`26.7
`14.7
`49.21
`25.3
`15.6
`9.0
`26.0
`42.8
`14.0
`
`25
`
`The results clearly show that these substituted glyco
`sides increase the corneal permeation of atenolol several
`fold over control.
`
`35
`
`EXAMPLE 11
`The following anti-allergic formulation was pre
`pared:
`
`wherein, R1 is a hydrophobic group including saturated
`and unsaturated aliphatic hydrocarbon groups which
`range from 8 to 28 carbons in length with 1 to 5 double
`bonds and said aliphatic hydrocarbon group can be a
`straight or branched chain and may be substituted by
`one or more aromatic, cycloaliphatic or hydrophilic
`groups;
`R2 is a group derived from any cyclic or acyclic
`saccharide containing 4-7 carbons;
`X is an integer from l-l0; and
`Z is an oxy (-0-), carbonyloxy
`
`phosphoryl
`
`thio
`(—S——-), or carboxamido
`
`Lodoxamide
`Dodecyl Maltoside
`Water
`pH was adjusted to 6.5
`
`% w/v
`
`0.1
`0.07
`q.s. to 100
`
`group where R; is covalently bound to such group.
`2. The method of cl