United States Patent (19)
`Horlington
`
`54 PHARMACEUTICAL COMPOSITIONS
`75) Inventor: Michael Horlington, Bishops
`Stortford, England
`73 Assignee: Smith and Nephew Associated
`Companies Limited, United Kingdom
`21 Appl. No.: 342,655
`22 Filed:
`Jan. 25, 1982
`
`Related U.S. Application Data
`Continuation-in-part of Ser. No. 287,356, Jul. 27, 1981.
`63
`Foreign Application Priority Data
`30
`Feb. 9, 1981 (GB) United Kingdom ................. 8103917
`Mar. 18, 1981 GB United Kingdom ................. 8108537
`51
`Int. Cl. .................... A61K 31/505; A61K 31/52
`52 U.S. C. ..................................... 424/253; 424/251
`58) Field of Search ................................ 424/251, 253
`56
`References Cited
`U.S. PATENT DOCUMENTS
`3,081,230 3/1963 Weinstock et al. ................. 424/25
`3,870,791 3/1975 Haddad et al. ..................... 424/25
`4,118,492 10/1978 Voelger et al. ..................... 424/251
`4,187,307 2/1980 Paris et al. .......................... 424/25
`4,285,947 8/1981 Higuchi et al. ..................... 424/25
`FOREIGN PATENT DOCUMENTS
`932256 7/1963 United Kingdom .
`1009477 11/1965 United Kingdom .
`
`11
`45
`
`4,425,346
`Jan. 10, 1984
`
`OTHER PUBLICATIONS
`Martindale-The Extra Pharmacopoeia, 27th Ed. (1977)
`pp. 569-570.
`Pteridines XII Structure-Activity Relationships of
`Some Pteridine Diuretics, Weinstock et al., J. Med.
`Chem. 1968. 11 573-579.
`2,4,7-Triamino-6-Ortho-Substituted Aryl Pteridines,
`A New Series of Potent Antimalarial Agents, Osdene et
`al., J. Med. Chem. 1967 10 pp. 431-433.
`Primary Examiner-Douglas W. Robinson
`Attorney, Agent, or Firm-Jacobs & Jacobs
`(57)
`ABSTRACT
`The present invention relates to pharmaceutical compo
`sitions adapted for topical administration to the eye and
`to their preparation and use. More specifically this in
`vention relates to anti-glaucoma ophthalmic composi
`tions containing tetraazabicyclic compounds of the
`formula:
`
`
`
`wherein R, R2, R3, X and Y are hereinafter defined and
`to the preparation and use of such compositions.
`
`61 Claims, No Drawings
`
`MYLAN INST. EXHIBIT 1063 PAGE 1
`
`MYLAN INST. EXHIBIT 1063 PAGE 1
`
`

`

`4,425,346
`
`1.
`PHARMACEUTICAL COMPOSITIONS
`This is a continuation-in-part of application U.S. Ser.
`No. 287,356 filed July 27, 1981 and entitled “Pharma
`ceutical Composition.'
`Glaucoma is a degenerative disease of the eye mani
`fest inter alia by an elevated intra-ocular pressure in the
`eye. Ocular hypertension, that is the condition of ele
`vated intra-ocular pressure, is believed by many author
`ities to represent an early phase in the onset of glau
`coma. One method of treatment of ocular hypertension
`and glaucoma is to administer to the subject a pharma
`cologically active compound capable of reducing the
`intra-ocular pressure. A number of compounds pres
`ently employed to treat ocular hypertension and glau
`coma are not entirely satisfactory due at least in part to
`side effects such as pupil contraction and the like.
`Clearly it would be desirable to provide an agent which
`could be applied topically to treat ocular hypertension
`20
`and glaucoma without an unacceptable level of such
`side effects. It has now been found that the optical ad
`ministration of tetraazabicyclic compounds to the eye
`can reduce the intra-ocular pressure therein without
`producing an unacceptable level of side effects such as
`25
`pupil constriction.
`It has now been found that the topical administration
`of other tetraazabicyclic compounds to the eye results
`in a lowering of intra-ocular pressure and that this oc
`curs without systemic side effects such as diuresis and
`30
`without local side effects such as pupil constriction.
`Accordingly the present invention provides a phar
`maceutical composition adapted for administration to
`the eye which composition contains a compound of the
`formula (I):
`-
`
`10
`
`15
`
`35
`
`2
`Weinstock et al in J. Med. Chem. 1963, 11573-79. This
`paper contains no suggestion that such compounds may
`be applied topically to the eye and no suggestion is
`made that such compounds may be useful in the treat
`ment of glaucoma.
`Compositions suitable for topical application to the
`eye containing 2,4,7-triamino-6-phenyl-pteridine are
`described in our co-pending U.S. applications, Ser. Nos.
`287,356 and 328,729 which are incorporated herein by
`cross-reference.
`When used herein the term lower means a group of 1,
`2 or 3 carbon atoms and most aptly refers to a group
`containing 1 carbon atom.
`In compounds of the formula (I) as hereinbefore de
`fined most aptly R3 is an amino group.
`In compounds of the formula (I) as hereinbefore de
`fined most aptly X=Y is a CR“=N group where R“is
`a hydrogen atom, a lower alkyl, an aryl or a carbox
`amido group.
`Thus favoured compositions contain a compound of
`formula (II):
`; : . . .
`. "
`
`
`
`(II)
`
`wherein either R5 and R6 are amino groups or one of R5
`and R6 is an amino group and the other is a hydrogen
`atom, a lower alkyl or an aryl group; and Ris a hydro
`gen atom, a lower alkyl, an aryl or a carboxamido;
`together with a carrier therefor.
`Particularly apt compositions contain a compound of
`formula (III):
`
`"Se N
`N s
`*sy J. N
`
`R3
`
`(I)
`
`40
`
`wherein X=Y is CR=N or N=CR where at least two
`of R, R, R2 and R3 are amino groups and the other
`groups of R, R, R2 and R3 are hydrogen atom, lower
`45
`alkyl, aryl, carboxamido or lower alkoxycarbonyl
`groups; together with a carrier therefor.
`Compounds of the formula (I) may be prepared in
`known manner, for example as in the following series of
`papers, (a) Spickett and Timmis, J. Chem. Soc. 1954,
`50
`2887-95; (b) Pachter and Nemeth, J. Org. Chem. 1963,
`28 1187–91; (c) Pachter, ibid 1963, 28 1191-96; (d)
`Pachter and Nemeth, ibid 1963 28 1203–06; (e) Wein
`stock et al, J. Med. Chem. 1968, 11 542-48; (f) Wein
`stock et al, ibid, 1968, 11 549–56; (g) Weinstock et al,
`55
`ibid 1968, 11 557-60; (h) Weinstock and Dunoff, ibid
`1968, 11 565-68; (i) Weinstock et al, ibid 1963, 11
`618-20; (j) Mallette et al, J. Am. Chem. Soc. 1947, 69
`1814. The diuretic properties of such when given orally
`to rats and dogs is described by Weinstock et al in J.
`60
`Med Chem. 1963, 11 573-79. However this paper con
`tains no suggestion that such pteridine derivatives may
`be applied topically to the eye and no suggestion is
`made that such pteridine derivatives may be useful in
`the treatment of glaucoma. The preparation of some
`65
`further compounds of the formula (I) is described in
`Graboyes et al, J. Med Chem. 1963 11 568-73. The
`diuretic properties of these compounds is described in
`
`(III)
`
`H2N a N
`
`N N
`
`I N I N
`.
`...
`NH,
`wherein R8 is a hydrogen atom, a lower alkyl, an aryl,
`a lower alkoxycarbonyl or carboxamido group and Ris
`a phenyl group or a phenyl group substituted by a fluo
`rine, chlorine or bromine atom or by a lower alkyl,
`lower alkoxy, amino, nitro, hydroxyl or sulphated hy
`droxyl group; together with a carrier therefor.
`Aptly R7 is a phenyl group optionally substituted in
`the 3 or 4 position.
`; .
`. . .
`.
`.
`.
`.
`Preferably R7 is a phenyl group.
`From the foregoing it will be appreciated that highly
`favoured compounds for use in this invention include
`2-phenyl-4,7-diamino-6-carboxamido-pteridine and the
`ethyl ester of 2-phenyl-4,7-diamino-pteridine-6-car
`boxylic acid.
`Particularly suitable compositions contain a com
`pound of formula (IV):
`
`MYLAN INST. EXHIBIT 1063 PAGE 2
`
`MYLAN INST. EXHIBIT 1063 PAGE 2
`
`

`

`3.
`
`N s-N".
`r N
`S. N I N
`NH2.
`
`R10
`
`m
`
`(IV)
`
`O
`
`15
`
`wherein R9 is a hydrogen atom, an amino group, a
`lower alkyl group or an aryl group and R10 is a hydro
`gen atom, a lower alkyl group, an aryl group or a car
`boxamido group; together with a carrier therefor.
`Aptly R9 is an amino group or a methyl group.
`Preferably R9 is an amino group.
`Aptly R10 is a phenyl group optionally substituted by
`a fluorine, chlorine or bromine atom or by a lower alkyl
`or in any position by lower alkoxyl, amino, nitro, hy
`droxyl or sulphated hydroxyl group.
`Preferably R10 is a phenyl group substituted as herein
`before described or a methyl group.
`From the foregoing it will be appreciated that highly
`preferred compounds for use in this invention include
`2,4,7-triamino-6-(4-nitrophenyl)-pteridine,
`2,4,-
`triamino-6-(4-hydroxyphenyl)-pteridine, the hemi-sul
`phate ester of 2,4,7-triamino-6-(4-hydroxyphenyl)-pteri
`dine and 2,4-diamino-6,7-dimethyl-pteridine.
`The preferred compound for use in this invention is
`the hemi-sulphate ester of 2,4,7-triamino-6-(4-hydroxy
`phenyl)-pteridine.
`A second group of suitable compounds are those of
`30
`formula (I) in which X=Y is N=CR namely com
`pounds of formula (V) or a pharmaceutically acceptable
`salt thereof.
`
`25
`
`4,425,346
`4.
`In order to be suitable for application to the eye the
`topical composition should be sterile, non-toxic and
`non-irritant to the eye.
`From the foregoing the skilled worker will appreci
`ate that this invention provides a sterile, non-irritant,
`non-toxic composition adapted for topical administra
`tion to the eye for the treatment of glaucoma which
`composition comprises a compound of formula (I) as
`hereinbefore described together with a carrier therefor.
`Suitable forms of the composition include aqueous
`solutions, aqueous suspensions, oily solutions, oily sus
`pensions, ointments, emulsions and sustained release
`implants. In general it is preferred to use aqueous solu
`tions or aqueous suspensions for compositions of this
`invention. Such aqueous forms preferably also contain
`an agent which increases the amount of the compound
`of formula (I) in suspension (a suspending agent) or
`solution (a solubilising agent) as described hereinafter.
`Most desirably the composition of this invention will
`be an aqueous solution.
`Normally and preferably aqueous solutions and sus
`pensions of the invention will contain tonicity adjusting
`agents, for example sodium chloride, potassium chlo
`ride, glycerol, propylene glycol, urea or dextrose to
`render the solution or suspension isotonic or substan
`tially isotonic with tear fluid, that is to say, to give the
`compositions a tonicity equivalent to an aqueous solu
`tion containing from 0.8 to 1.1% of a sodium chloride
`and most suitably 0.9% sodium chloride. The use of
`sodium chloride or other ionic tonicity agents may
`render some solutions and suspensions unstable. In these
`cases it is preferred that a non-ionic tonicity adjusting
`agent such as glycerol or propylene glycol is used.
`Aqueous suspensions of the invention will suitably
`contain from 0.01 to 5% of the compound, more suit
`ably will contain from 0.05 to 2.5% of the compound
`and preferably from 0.1 to 1% for example 0.5% (9%
`terms when used herein are expressed on a wt/v basis
`unles shown otherwise).
`Aqueous solutions of the invention will contain an
`amount of the compound which will depend on the
`solubility of the specific compound used. Normally
`aqueous solutions may contain from 0.01 to 2% of the
`compound.
`It is normally preferred in the treatment of glaucoma
`to employ an aqueous solution. Favourably the aqueous
`solutions of this invention will contain a solubilising
`agent such as polyvinyl pyrrollidone, polyalkylene gly
`col, non-ionic surfactants or a polyacrylic acid which
`has been lightly cross-linked with triallyl sucrose in
`combination with a non-ionic surfactant. An apt solubi
`lising agent is polyvinyl pyrrolidone. Suitable polyvinyl
`pyrrollidones are those with a number average molecu
`lar weight below 40,000, more suitable are those with a
`number average molecular weight below 5000 and a
`particularly preferred polyvinyl pyrrolidone is one hav
`ing a number average molecular weight of 2,500. Such
`a polyvinyl pyrrolidone is exemplified by Kollidon
`12PF (trade mark of BASF). Suitably the amount of
`polyvinyl pyrrollidone present is from 0.5 to 55%, pref.
`erably is from 15 to 52% and most preferably is from 20
`to 50%.
`Alternatively the compounds useful in the present
`invention may be solubilised by employing a class of
`agents that neither unduly increases the viscosity of the
`solution nor unduly increases the surfactant properties
`of the solution. This class of agents which may be thus
`employed are water soluble xanthine derivatives.
`
`35
`
`(V)
`
`R11
`
`R13
`
`where at least two of R12, R13 and R1 are amino groups
`and the other group is a hydrogen atom or a phenyl
`group or a phenyl group optionally substituted by a
`fluorine, chlorine or bromine atom or by a lower alkyl,
`45
`lower alkoxy, amino, nitro, hydroxyl or a sulphated
`hydroxyl group and where R1 is a hydrogen atom, a
`phenyl group, or a phenyl group optionally substituted
`by a fluorine, chlorine or bromine atom or by a lower
`alkyl, lower alkoxyl, amino, nitro, hydroxyl or sul
`phated hydroxyl group.
`Preferably R12 and R13 are both amino groups.
`Preferably R1 is an amino group, a hydrogen atom or
`a phenyl group.
`Preferably R1 is an amino group.
`Aptly R1 is a hydrogen atom or a phenyl group.
`Preferably R1 is a phenyl group.
`From the foregoing it will be appreciated that highly
`favoured compounds for use in this invention include
`2,4,7-triamino-5-phenyl-pyrimido (4,5-d) pyrimidine.
`60
`Pharmaceutically acceptable salts of compounds of
`formula (V) include those with acids such as hydrochlo
`ric, sulphuric, orthophosphoric, acetic, gluconic, glu
`tamic and lactic acids.
`From the foregoing it will be appreciated that highly
`favoured compounds for use in this invention include
`2,4,7-triamino-5-phenyl-pyrimido (4,5-d) pyridinium
`orthophosphate.
`
`50
`
`55
`
`65
`
`MYLAN INST. EXHIBIT 1063 PAGE 3
`
`MYLAN INST. EXHIBIT 1063 PAGE 3
`
`

`

`15
`
`20
`
`4,425,346
`5
`6
`Thus in a more favoured aspect this invention pro
`like. The preferred quaternary ammonium compound is
`vides a sterile, non-irritant, non-toxic aqueous solution
`benzalkonium chloride. Suitably the amount of quater
`adapted for topical application to the eye for the treat
`nary ammonium compound present is from 0.005 to
`ment of glaucoma which composition comprises a com
`0.04%, more suitably is from 0.0075 to 0.025% and
`pound of formula (I) as hereinbefore described and a
`preferably from 0.01 to 0.02% of the composition.
`solubilising xanthine and a carrier therefor.
`In aqueous suspensions of compounds of formula (I)
`The solubilising xanthine is normally a dimethylxan
`the compound will be dispersed evenly throughout.
`thine which optionally may be substituted by a hydro
`The compound will be in finely divided form. In this
`philic group or (less preferably) a methyl group. Most
`state of subdivision of the compound 99% of the parti
`aptly the xanthine is a 1,3-dimethyl xanthine unsubsti
`cles are less than 30 microns in diameter and 90% are
`10
`tuted or substituted at the 7-position by a methyl group
`less than 10 microns in diameter. Most aptly the parti
`or more aptly substituted at the 7-position by a hydro
`cles will have diameters in the range of 1 to 5 microns.
`philic group such as a 2-hydroxypropyl group (thereby
`Generally the compositions of this invention will
`proxyphylline), a 2,3-dihydroxypropyl group (thereby
`contain a suspending agent for the finely divided com
`dyphylline), a 2-hydroxyethyl group (thereby etophyl
`pound of formula (I). Suitable suspending agents in
`line), a carboxymethyl group (thereby theophyllinyla
`clude cellulosic or polysaccharide derivatives (such as
`cetic acid), an N,N-diethylaminoethyl group (thereby
`hydroxyethyl cellulose, hydroxypropyl cellulose, car
`etamiphylline) or the like. Less favourably the xanthine
`boxymethyl cellulose or xanthan gum) or water-soluble
`is a 3,7-dimethylxanthine unsubstituted or substituted at
`polymers (such as polyvinyl alcohol or polyvinyl pyr
`the 1-position for example pentoxyphylline.
`rolidone or a polyacrylic acid lightly cross-linked with
`Particularly apt solubilising xanthines are caffeine,
`triallyl sucrose). A most favoured suspending agent is a
`proxyphylline and dyphylline. These agents may be
`polyvinyl pyrrolidone as hereinbefore described. Suit
`present in any convenient solubilising amount, for ex
`ably the suspension will contain from 5 to 55% of the
`ample from 0.1 to 10% (or up to the solubility limit),
`polymer and preferably 20 to 50% of the polymer.
`more usually from 1 to 8%. Preferably the solution will
`A second favoured suspending agent is a hydroxy
`25
`contain from 2 to 7.5% of proxyphylline or diphylline.
`ethyl cellulose. Suitable hydroxyethyl celluloses are
`The xanthine solubilising agent may be present as the
`available as the Natrosols (trade mark of Hercules Inc.)
`sole solubilising agent or less aptly other solubilising
`and the Cellosizes (trade mark of Union Carbide Corp.).
`agents such as those hereinbefore described may also be
`A 2% solution of suitable polymers at 25 C. has a vis
`included.
`cosity of from 4500 to 6000 cps when measured on a
`30
`The use of solubilising agents is particularly advanta
`Brookfield apparatus. A preferred hydroxyethyl cellu
`geous for the preparation of solutions of the sparingly
`lose is available as Natrosol 250M. Suitably the suspen
`soluble compounds
`sion will contain from 0.1 to 10% of the cellulose, more
`It is desirable to provide aqueous solutions that are
`suitably 0.25 to 2.5% of the cellulose and preferably 0.5
`generally at a pH of not less than 4.5, more aptly not less
`to 1.0% of the cellulose.
`35
`than 6 and preferably at a pH of approximate neutrality
`Generally the aqueous suspension of the present in
`that is pH 7. The pH of such solutions should also not in
`vention will contain a surface active agent to encourage
`general be greater than 8.5 and preferably not greater
`wetting of the surface of the particles of the finely di
`than pH 8. It has been found that such solutions may be
`vided compound by water, thus aiding even dispersion
`prepared by using solubilising agents such as a xanthine.
`throughout the suspension. Favoured surface active
`If the solubilising agent is a xanthine a buffering agent
`agents are polyoxyethylated sorbitan fatty acid esters
`may be present to maintain the pH from 5.0 to 8.0 and
`(commonly called Tweens) or polyoxypropylene
`preferably 6.0 to 7.5 for example pH 7. Suitable buffer
`polyoxyethylene diol block copolymers (commonly
`ing agents include mixtures of potassium dihydrogen
`called Pluronics). Particularly preferred is a polyoxy
`phosphate and disodium hydrogen phosphate and other
`propylene-polyoxyethylene diol block copolymer of
`45
`systems known to provide solutions having such pH
`molecular weight 2900, having 40% of ethylene oxide
`values.
`units in the polymer, known as Pluronic L64. Suitably
`In certain circumstances it may be preferred to use a
`the amount of surface active agent present is from 0.01
`pharmaceutically acceptable salt of a compound of
`to 5% and preferably from 0.02 to 0.2%.
`formula (I). Such salts include those with hydrochloric,
`Generally the aqueous suspensions will contain a
`50
`sulphuric, orthophosphoric, acetic, gluconic, glutamic
`ophthalmically acceptable preservative or bacterial
`and lactic acids. In general these salts are prepared in
`agent. Such agents are subject to the same consider
`situ and no not exist as true salts outside the formulation;
`ations of compatibility as described for aqueous solu
`that is the solutions of compounds of formula (I) may be
`tions above. Suitable bacterial agents include phenyle
`rendered slightly acidic using such acids so that a salt is
`thanol, phenoxyethanol, chlorbutanol or thiomersal. A
`55
`notionally formed.
`particularly preferred preservative is a combination of
`Normally and preferably aqueous solutions of the
`phenylethanol in an amount from 0.25 to 0.75% and
`invention will contain an ophthalmically acceptable
`thiomersal in an amount from 0.005 to 0.02%. A pre
`preservative to maintain the sterility of the solutions
`ferred combination is 0.5% phenylethanol with 0.01%
`during use. It is known in the art that certain preserva
`thiomersal.
`60
`tives are affected by the presence of surface active
`The pH values of the aqueous suspensions will be
`agents, viscosity increasing agents and the like and natu
`adjusted in a similar manner to that described hereinbe
`rally such agents will be selected to be mutually com
`fore with respect to aqueous solutions.
`patible in the conventional manner. Particularly apt
`Oily suspensions of compounds of formula (I) include
`ophthalmically acceptable preservatives are antibacteri
`suspensions in solvents such as castor oil and such com
`65
`ally effective quaternary ammonium compounds such
`positions will suitably contain from 0.01 to 5% of finely
`as benzalkonium salts, for example the chloride, benze
`divided compound, more favourably 0.02 to 0.75% and
`thonium chloride, cetyl pyrridinium chloride and the
`most preferably 0.05 to 0.5% of the compound.
`
`MYLAN INST. EXHIBIT 1063 PAGE 4
`
`MYLAN INST. EXHIBIT 1063 PAGE 4
`
`

`

`15
`
`30
`
`35
`
`0.5% w/v.
`50.0% w/v.
`to 100.0 ml.
`
`4,425,346
`8
`7
`tion are conventionally pre-sterilised and filled under
`Ointments in accordance with the invention will
`comprise ointment bases suitable for topical application
`aseptic conditions with the sterile composition using
`conventional metering pumps capable of delivering
`to the eye and contain suitably from 0.01 to 5% of the
`finely divided compound of formula (I).
`from 1 to 20 mls each filling cycle.
`Ocular implants may comprise a compound of for- 5
`. Alternatively for those compositions which are in the
`mula (I) in a finely divided form in suitable soluble or
`form of an aqueous solution, the solution may be formed
`insoluble material. Soluble forms for example where the
`by dissolving its components in distilled water and then
`finely divided compound is suspended in a polyvinyl
`sterilising the solution either by filtration through a 0.22
`alcohol film, dissolve and so do not need to be removed
`micron filter or by subjecting it to heat, for example by
`later, whereas the insoluble forms are removed from the 10
`autoclaving at 116 C. for 30 minutes at 10 psi pressure.
`eye after the medicament has diffused from the form.
`The sterile solution may be aseptically filled into pre
`Suitably the ocular implant will contain from 0.01 to
`sterilised eye-dropper bottles in a conventional manner.
`5% of finely divided compound of formula (I).
`-
`The compositions of this invention are non-irritant,
`The present invention also provides a method for
`that is they do not cause unacceptable irritation to the
`reducing the intra-ocular pressure in the eye which
`eye when applied topically. Most aptly the composi
`comprises topically administering a compound of for
`tions are bland upon application.
`-
`mula (I) to the surface of the eye in an amount effective
`The following Examples illustrate this invention:
`to reduce the intra-ocular pressure therein. The com
`EXAMPLE 1
`pound of formula (I) will be present in a composition as
`2,4,7-Triamino-6-(2-bromophenyl)-pteridine suspension
`herein-described.
`20
`Further the present invention provides a unit dose of
`A suspension of 2,4,7-triamino-6-(2-bromophenyl)-
`a liquid topical composition of the invention having a
`pteridine was prepared as follows:
`volume from 0.01 to 0.08 ml (i.e. a drop of 10 to 80
`microliters) and containing from 1 microgramme to 1.6
`mg of a compound of formula (I) and more usually 0.02 25
`to 0.05 ml and containing 2 microgrammes to 1 mg. of
`the compound. .
`.
`. .
`-
`The compositions of this invention are most aptly
`provided in a multidose container from which drops
`may be dispensed into the eye. Such containers are well
`known in the art for dispensing of liquid drops into the
`eye and such conventional containers may be em
`ployed. Most aptly such containers are adapted to hold
`1 to 20 mls, more usually 2 to 12 mls and preferably 3 to
`10 mls.
`A favoured aspect of the present invention therefore
`comprises a container adapted to deliver drops of an
`aqueous solution or an aqueous suspension of this inven
`tion as hereinbefore described. A preferred container
`comprises a glass bottle having a screw cap. This screw
`40
`cap being replaced by a screw cap carrying a drop
`forming portion when in use. A second preferred con
`tainer comprises a plastic bottle, for example of a low
`density polyethylene, having an integral dispensing tip
`covered by a screw cap. Such preferred containers are
`45
`adapted to hold 1 to 20 mls of the composition and
`preferably 3 to 10 mls of the composition, for example 5,
`7.5 or 10 mls. A third preferred type of container
`adapted to deliver drops holding from 1 to 2.5 mls of
`composition is made from polypropylene or other heat 50
`stable material whereby the whole package may be
`filled and sealed prior to sterilisation by autoclaving.
`The compositions of the present invention may be
`prepared by conventional means of mixing and blend
`55
`1ng.
`Sterile compositions may be conveniently prepared
`by (a) sterilising the finely divided compound of for
`mula () by dry heat over a period of time at elevated
`temperature, for example by heating to 160 for 1 hour,
`allowing the powder to cool and storing aseptically, (b) 60
`sterilising the liquid components of the composition by
`either filtration through a 0.22 micron cellulose ester
`membrane or subjecting the solution to heat and pres
`sure such as autoclaving at 116 C. for 30 minutes under
`10 psi pressure. The two sterile components are com- 65
`bined in the desired proportions under aseptic condi
`tions and filled into sterile containers. The containers
`for delivering drops of sterile compositions of the inven
`
`2,4,7-Triamino-6-(2-bromophenyl)-pteridine
`"Polyvinyl pyrrolidone (molecular weight 2,500)
`Distilled water
`The polyvinyl pyrrolidone was Kollidon PF12.
`The polyvinyl pyrrolidone was dissolved in water (40
`ml) and the 2,4,7-triamino-6-(2-bromophenyl)-pteridine
`suspended in this solution with stirring. The volume of
`the suspension was adjusted to 100 ml with distilled
`water with stirring.
`:
`The effect of this suspension on the ocular tension in
`the eyes of rabbits (New Zealand. White/Male) was
`tested as follows. 50 microliters of the suspension was
`applied topically to both treated and untreated eyes was
`measured at 1, 2, 4 and 5 hours after the application of
`the suspension. The mean maximum fall in ocular ten
`sion in the treated eye of four rabbits was 3.4 mmHg
`(approximately) at 1 hour, in the untreated eye the mean
`maximum fall was 1.6 mmHg (approximately) at 1 hour.
`. The pupil diameter of both treated and untreated eye
`remained unchanged over the test period.
`EXAMPLE 2.
`2,4,7-Triamino-5-phenylpyrimido(4,5-d) pyrimidone
`solution
`A solution of 2,4,7-triamino-5-phenylpyrimido (4,5-
`d)pyrimidine was formulated as follows:
`
`0.5% wav
`
`50.0% w/v.
`to 100.0 ml.
`
`2,4,7-Triamino-5-phenyl
`pyrimido(4,5-d)pyrimidine
`"Polyvinyl Pyrrolidone (molecular weight 2,500)
`Distilled water
`*The polyvinyl pyrrolidone was Kollidon PF12.
`The polyvinyl pyrrolidone was dissolved in water (40
`ml) and the 2,4,7-triamino-5-phenylpyrimido(4,5-d) py
`rimidine was added to this solution and stirred until
`dissolved. The volume of the solution was adjusted to
`100 ml with distilled water.
`The effect of this solution on the ocular tension in the
`eyes of rabbits (New Zealand White/Male) was tested
`as described in Example 1. The mean maximum fall in
`ocular tension in the treated eyes was 3.1 mmHg (ap
`
`MYLAN INST. EXHIBIT 1063 PAGE 5
`
`MYLAN INST. EXHIBIT 1063 PAGE 5
`
`

`

`4,425,346
`10
`proximately) occuring approximately 2 hours after in
`The suspension was prepared in a similar manner to
`stillation, the untreated eyes showed a mean maximum
`that of Example 1.
`fall of 1.6 mmHg (approximately also occurring approx
`The effect of this solution on the ocular tension in the
`imately 2 hours after instillation.
`eyes of rabbits (New Zealand White/Male) was tested
`The pupil diameter of both treated and untreated eye
`as described in Example 1. The mean maximum fall in
`remained unchanged over the test period.
`ocular tension in the treated eyes was 0.9 mmHg. The
`untreated eyes showed no corresponding fall in ocular
`EXAMPLE 3
`tension.
`2,4-Diamino-6,7-dimethylpteridine suspension
`The pupil diameter of both treated and untreated eyes
`A suspension of 2,4-diamino-6,7-dimethylpteridine
`remained unchanged over the test period.
`was prepared as follows:
`EXAMPLE 6
`2,4,7-Triamino-5-phenylpyrimido-(4,5-d)pyrimidinium
`orthophosphate Solution
`
`O
`
`2,4-Diamino-6,7-dimethylpteridine
`"Polyvinyl pyrrolidone (Mol. wit, 2,500)
`Distilled water
`The polyvinyl pyrrolidone was Kollidon TF12
`
`0.5% w/v.
`50.0% w/v
`to 100.0 m.
`
`15
`
`The polyvinyl pyrrolidone was dissolved in water (40
`ml) and the 2,4-diamino-6,7-dimethylpteridine sus
`pended in this solution with stirring. The volume of the
`suspension was adjusted to 100 ml with distilled water
`with stirring.
`The effect of this suspension on the ocular tension in
`25
`the eyes of rabbits (New Zealand White/Male) was
`tested as described in Example 1. The mean maximum
`pressure fall in ocular tension in the treated eye for four
`rabbits was 1.6 mmHg. In the untreated eye no signifi
`cant fall in ocular tension was shown.
`The pupil diameter of both treated and untreated eyes
`remained unchanged over the test period.
`EXAMPLE 4
`2-Phenyl-4,7-diamino-6-carboxamidopteridine
`A solution of 2-phenyl-4,7-diamino-6-carboxamido
`pteridine was prepared as follows:
`
`35
`
`30
`
`2-Phenyl-4,7-diamino-6-carboxamidopteridine
`"Polyvinyl pyrrolidone (Mol. Wt. 2,500)
`Distilled water
`"The polyvinyl pyrrolidone was Kollidon PF12
`
`0.5% w/v.
`50.0% w/v.
`to 100.0 ml.
`
`40
`
`2,4,7-Triamino-5-phenylpyrimido-(4,5-d)
`pyrimidinium orthophosphate
`Polyvinyl pyrrolidone (Mol. wt. 2,500)
`Distilled water
`
`0.5% w/v.
`50.0% w/v.
`to 100.00 m.
`
`A solution of 2,4,7-triamino-5-phenylpyrimido-(4,5-
`d) pyrimidinium phosphate was prepared, in a similar
`manner to Example 2.
`The effect of this solution on the ocular tension in the
`eyes of rabbits (New Zealand White/Male) was tested
`as described in Example 1. The mean maximum fall in
`ocular tension in the treated eyes was 2.6 mmHg. The
`untreated eyes showed no fall in ocular tension.
`The pupil diameter of both treated and untreated eyes
`remained unchanged over the test period.
`-
`EXAMPLE 7
`" 2,4,7-Triamino-6-(4-hydroxyphenyl)-pteridine
`suspension
`;
`:
`, ,
`
`A suspension of 2,4,7-triamino-6-(4-hydroxyphenyl)-
`pteridine is prepared as follows:
`. .
`
`0.5%
`2,4,7-Triamino-6-(4-hydroxyphenyl)-pteridine)
`50.0%
`Polyvinyl pyrrolidone (Mol. wt. 2,500)
`2.0%
`Propylene glycol
`0.5%
`Phenyl ethanol
`Sodium hydroxide solution to adjust pH to 7 as required
`Distilled water
`to 100 ml.
`
`50
`
`The polyvinyl pyrrolidone was dissolved in water (40
`45
`ml) and the 2-phenyl-4,7-diamino-6-carboxamidopteri
`dine was added and stirred until dissolved. The volume
`of the solution was adjusted to 100 ml with distilled
`Water.
`The effect of this solution on the ocular tension in the
`eyes of rabbits (New Zealand White/Male) was tested
`as described in Example 1. The mean maximum fall in
`ocular tension in the treated eye was 3.6 mmHg. The
`untreated eyes showed a mean maximum fall of 1.6
`mmHg.
`The pupil diameter of both treated and untreated eyes
`remained unchanged over the test period.
`EXAMPLE 5
`2-Phenyl-4,7-diamino-6-ethoxycarbonyl pteridine
`suspension
`
`55
`
`60
`
`2-Phenyl-4,7-diamino-6-ethoxycarbonyl
`pteridine
`50.00% w/v.
`"Polyvinyl pyrrolidone (Mol. wt. 2,500)
`to 100.0 ml.
`Distilled water
`"The polyvinyl pyrrolidone was Kollidon PF12 (Registered Trademark of BASF).
`
`0.50% w/v .
`
`65
`
`The finely divided 2,4,7-triamino-6-(4-hydroxy
`phenyl)-pteridine powder is heated at 160° C. for 1
`hour, sufficient for dry heat sterilisation, allowed to
`cool and stored under aseptic conditions.
`The polyvinyl pyrrolidone, propylene glycol and
`phenyl ethanol are dissolved in distilled water (40 ml).
`The pH of this solution is adjusted to pH 7 with sodium
`hydroxide solution and the volume finally adjusted to
`100 ml by addition of further distilled water. This solu
`tion is then sterilised by autoclaving at 116° C. for 30
`minutes and then allowed to cool.
`-
`The remaining procedures are carried out in aseptic
`conditions under a laminar flow hood.
`The sterile powder is triturated in a pestle and mortar
`with a portion of the sterile solution. This suspension is
`quantitatively added back to the bulk of the sterile solu
`tion and the whole mixed thoroughly.
`The suspension is then aseptically filled into sterile
`