CA2723144A1 - Ophthalmic device, and method of use thereof, for increasing ocular boundary lubrication - Google Patents

Abstract

The present invention provides an ophthalmic device, and method of use thereof, for an individual wearing an ophthalmic lens to increase ocular surface boundary lubrication. The invention device comprises an ophthalmic lens and a sacrificial mechanism disposed on the ophthalmic lens, wherein the sacrificial mechanism comprises a plurality of surface bound receptors, such as PRG4, hyaluronic acid, and DNA aptamers, that reversibly bound to a lubricating composition comprising a gel forming agent, a surfactant, or a combination thereof, effectively inhibiting or preventing protein and lipid adsorption on the surface of the lens, and mitigate shear stress and reduce the friction between the lens and the ocular surface of the individual in need.

Description

OPHTHALMIC DEVICE, AND METHOD OF USE THEREOF, FOR INCREASING OCULAR BOUNDARY LUBRICATION
CROSS-REFERENC TO RELATED APPLICATIONS

1,01] This patent application claims prioritav benefit of U .S. Provisional Application No.
61/051,11.2 filed. 'Ian, 7, 2 j0lia which is i:ncorpo-rated herein by reference.

FIELD O.l I HE INVENTION

It012l The present invention relates to an ophthalmic device for the management of ocular lubrication in the presence of an ophthalmic lens.

BACKGROUND
I'M31 The proteoglycan 4 pa -4:) gene encodes for highly glycosylated proteins termed megt,A-aryocyte stimulating,, factor (NISF). lubir-icin. and superficial zone.
protein (SZP) (1).
These .trac;le rule s are collectively referred to as PRG4 or F RG4 proteins.
PRG4 is pre-sent in synovial fluid and at the surthce of synoviuni (2), tendon (3) end m eniscus (4),111d is suspected as being an important component for health synovial .joints. See, e.g., (`). (0).

1O04l In tissues such as syno ial,joints, physicoehenucal modes of lubrication have been classified as fluid film or boundary.. The operati ve lubricat.iou mtiodes deÃaend on the normal and tang ntial forces on the articulating tissues., on the :relative rate of tangential motion between these surface, and on the time history of boÃ1h, loading, and motion. The friction coctticient, p, provides a quantitative aarea ur . and is defined as the ratio of tangential friction force to the normal force. One type of t ).aid-med iaated lubrication mode is h drostatic. At the onset of loading and t =pically for a prolonged duration, the interstitial fluid within cartilage becomes pressurized, due to the hiphasic nature cif the tissue; fluid may also be forced into the aasperit:ies between articular surf aces through a weeping mechanism. Pressurized interstitiail fluid and trapped lubricant pools ma th reÃÃ?re contribute significantly to the bearing of normal load with little resistance to shear force, thc:ilitaatin, a very kn p. Also, at the onsa of loading and/or motion, squeeze film, hydrodynamic, and e.iastohvdrodynan:aic types of fluid. film) .lubrication occur, with pressurization, motion,, and deforraaation acting to drive viscous .lubricant fix-?m and,or through the ;aap between two sagafhces in relative ramot oar.

lttrt l The,, relevant e:; tent to which fluid prÃ;ssure/filer. versus boundary lubrication occurs classically depends on a number of factors (13). When lubricant film can l'1Ã?
between the cc_rnfiorfning sliding surfaces, which can deform Blast call. elastohydrodv namic lubrication occurs. Pressure, surface raughness, and relative sliding velocity determine when full fluid lubrication begins to break down and the lubrication enters new .regimes. As velocity decreases :further. lubricant films adherent to the articulating surfaces begin to contribute and a mixed re<gime. of lubrication occurs. If the velocity decreases even further and only au rr.ltra--thin lubricant layer composed. of a tew r o.lecules remain boundary luhr-icatia:r7 occurs.. boundary mode of lubrication is therefore indicated by a friction coefficient (ratio of the measured frictional force E cetweera two contacting surfaces in relative motion to the applied normal force) during steady sliding Wing invariant w-Ot.h f tc:tors that influence formation of to fluid film, such as relative sliding velocity and axial load (14).
For articular cartilage, it has been concluded boundary lubrication is certain to occur.
although complemented by fluid pressurization and other mechanisms (15-la).

(ttt061 In boundasi lubrication, load is supported by surface-to-surface contact, and the associated fictional properties are determined by lubricant surface:
molecules. This mode.
has been proposed to be important because the opposing cartilage layers make contact over 10% of the total area, and this may be vvla.ere. most of the friction occurs (19).
Furthermore., with increasing loading, tim and dissipation of hydrostatic pressure.
lub:.6c tnt-coated surfaces bear an i:ncreasing.lty higher portion of the load, relative to pressurized fluid and conseclrr.cntly this mode can b, com :inc.rÃa`ingl dominant (1-1, 20).
Boundary lubrication, in essence, mitigates stick-slip (13), and is therefore.. manifest as decreased resistance both to steady motion and the start-up of motion. The latter situation is relevant to load bearing articulating surf-aces after prolonged compressive loading (e.g..

2: sitting or standing in vivo) (21). Typical wear pattenis of cartilage surfaces (22) also suggest that boundary lubrication of articular cartilage: is critical to the protection and maintenance of the articular sat-ace stnwture.

10071 With increasing loading time and dissipation of hydrostatic pressure.
lubricant-cc-rated surfaces bear an increasingly higher portion of the load relative to pressurized fluid..
and consequently, p can become increasingly dominated by this mode of lubrication, A
boundary erode of lubrication is indicated by values of p duringr steady sliding being

3 PCT/US2009/043018 inva.riant i it:h factors that influence fbn .ation of a fluid film, such as relative sliding velocity and axial load. Boundary lubrication, in essence, mitigates stick-slip, and is therefore manifest as decreased resistance both to steady motion and the start-up of motion.

ltaÃrsl The precise mechanisms of boundary lubrication at biological interfaces are currently unknown. However, proteo4glycan 4 (:P.R(=4) may play a critical role as a houndaarv lubricant in articulating Joints. This secreted glycoprotein is thought to protect, cartilaginous surfaces against frictional force-,;, cell adhesion and protea deposition, Various native and recombinant lubricira proteins and iscoforrns have been isolated and 1.0 characterized. For instance. U.S. Patent Nos. 5326,558-, 6,433,142-, 7,03022:3. and 7,361,738 disclose aa. family of human me.gakkaaiyocyte stiniulating. factors (.MSFs) and pharmaceutical compositions containing one or more such MSFs for treating disease states or disorders, such. as a deficiency of platelets. U.S. Patent loos. 6,960,562 and 6,743,774 also disclose aa. lubricating polyfpeptide, trihonectin, co Trp:rising a substantially pure fragments of MSF., and .methods of lubricating joints or other tissues by administering triboraectira sa: stern icalls or directly to tissues.

1009] A challenge to boundary lubrication is the presence of inflammation in surrounding tissues, as well as increased protease levels in the sv'novial fluid. Loss of the boundary-lubric atinlg ability of synovial fluid after in jury is associated ` ith damage to the articular cartilage matrix. This can be attributed to i.n.flatnniato ' processes resulting from the injury, particularly in the early phases. Another challenge to bouadarr lubrication is a sex steroid inabalaance, especially in arthritic disorders such as rheumatoid aatha=atis. Sex steroids are involved in the pathogenesis and regulation of inflammation in rheumatoid arthritis, a disease characterized by chronic inflammatory synovitis..ndrogens sup rcss, whereas estrogens promote, inflammatory proc..sses. Consequently, the relative levels of androgens and estrogens in the st novi al environment are extremely important in gression of inflammation (:7 8. 23). Various androgen compounds determining the pro veduc c. the magnitude of lymphocyte infiltration in lacrimal tissue. See, e.g;., U.S. Patent Nos 4i,620,921, 5_688_76553i20v921. and 6,107 2.8 }.

loolol Engineering of contact lens surfaces have traditionally fbcuscd on increasing oxygen transport. Recent advances in contact lens chemistries have also focused ou

4 increasing water content and .hvdrophilicits; to inhibit protein deposition on the. lens surface. Protein deposition on the poteriorinn:r surface of the Contact lens surfaces has been implicated as a causative factor in the corneal abrasions and mechanical trauma associated with contact lens Wear.

l00Ãt1 Advances in `il.icone latclro e..I aaaateai~als haatie aired populaarit due tea their rabilit to reduce protein. absorption through increased hydrophilicit Examples include I,otrattlcon A. ( ' -cl.ina.ctl~ lacr~ l~an3iclc. tria3.aethvlsiloxyy silan ;
and silox ane monomer, CIBA Vision. a.k.a. Focus NIGHT & DAY., 24`#.k water contetnt_ l7_? D It +0:, transu issibil.ata, ), Lotrafilcon B l d.ia ae tla~ lacy l.taYaiale.
trlinctlhvlsilcoxy `ilaac and 1.0 siloxane monomer, CIBA Vision, a.k.a. O2 Optix, 33% pater content. 138 Dk/t Ã), transmissibihtc. 1, I3alaf loon (N-vinyl pti rrolidonc, this tz aaaatla l tlo:s ilt l} prop l iai. t earl nn rte.. N-carbox vin' I ester, laol a diraae t1a silcaxe }
di(si1o'lbutarml) his( in 1 ca bamate). Bausch Lomb; a.k.a. PureV i ion, 36"/%x water contents 101 DIvI 02 transmissibility } Galvfilcon A (monofunctionai polvdimethylsilo .came. N,,N-l 5 dia r .ths .cn ltazmdu, Bolt 2rh ydaoti' ctl-ay 1 nwth acry late;:, si(o.\.aanc monomer, pol's r ns>l pyrrolidon.e, etlhvlt negiycol d.ianethacas late, Johnson Johnson Vision Care.
ra.k.a.
AC`UV UlE Advance. 47% w vate.r content, 86 Dk/t C transmissib: laity )_ 1 tafilcoa3 AA (poly-2-hvdroxyethv1 tne:thacr vlate_ methacrviic acid. Johnson & Johnson Vision Care, a.k.a.
ACl?VU12', water content, 21 Dk/t 0.T transmissibility), hi addition to the material 20 choices. these silicone. hydro el lenses are also manufactured with an additional treatment steps to improve hydrophi.licity. For example, Lotrafilcon A use plasma coating..
Balafilcon A makes use of a plasma oxidation process, and A(CU f `I Ad ance l :.tries include poly vir N pvrrolhdorte as an internal letting agent I: 5J. Plasma treatments are known to fade: and lose eMicacv over time.

25 100121 Protein adsorption at contact lens surf ces is commonly attributed to human albumin and lvsozyme, two of the most abundant proteins in the tear film.
Conflicting results have been reported regarding the i rrtea coati apt, 1az d.rcpplfc hicita charge.j pore size and surface roughness, Because the isoelectric points of albumin and lvs`az-vmo are. on opposite ends of the pH of human tear. the minimum protein adsorption seems to occur 30 when charge, water content and hvdrophilicity are properly balanced. Lower water content materials tend to bind albumin w to ile higher water content mate;ri als tend to bind s lv ozv3rao 124]. I oaeaa aaaaaaaa et_ al [241 also iraolrs anted that silicon l adro? els exhibit both hydrophobic and hydrophilic domains:. and follosving evaporation, chain rotation k r ;es tend to expose hydrophobic domains to the air. thereby increasing the chance for dry spots. Polar lipids nati also bind to hdroplhilic regions on the lens surface, resulting in exposed hydrophobic tails, a. hich may also) promote dry' spots, 1(1)131 yperosnaolaarity is a common result of contact lens wear. Those. with a reduced quantity or quality of lipid production tend to exhibit drastically less stable tear :films, and the presence of a contact lens may exacerbate the instability. This leads to a faster evaporation of the tear f-ilaam, and a. eoncentmtic-on of the tears o sver the a sc:ular surface.

SUMMARY OF THE INVENTION

[04)14) The. present invention provides, in various embodiments, an ophthalmic device for the management of ocular lubrication in the presence of an ophthalmic lens.
Given the relationship between osmotic pressu=re and the electromechanical interactions . -ithin charged molecules, the present .invention provides for the method of ma nag.ing decreased 14; ocular boundary lubrication in ophthalmic lens wear by modulating hypcrosinolarity at the (_ocular- surface. In certain instaances, by interrupting the feedback mechanisms which promote hyperosmolarity, the .integration of a Sacrificial Mechanism .into the pre-- aaa.dr'or postocular ophthalmic lens reduces the static and kinetic friction coefficient at the ocular surface during an eyelid blink. In some instances, over time., the reduction in shear stress alleviates by perosmolarity driven by the gain of this feedback rarechanisrn.

lool5l Described in certain embodiments of the present invention is a sacrificial mechanism disposed on at least a portion of the inner surface in an amount effective to provide ocular boundary lubrication in an individual wearing the ophthalmic lens. In one embodiment of the current invention. the sacrificial mechanism Comprises a plurality of 2.5 lubricating surface bound receptors, such as plurality of PRG4 a pluraa.litt of PRG4 molecules). In this embodiment. the lubricating surface bound receptor (e.g., PRG4) is allowed to interact with eudogenous proteins and proteoglycaris within the tear film to facilitate activation of the sacrificial mechanism. In. sonic instances, this interaction prevents or inhibits protein or lipid adsorption at the lens surface, reduce dry spots on the W lens, and r:duce t . friction between tlae lens and the oeorlaar ;surface..
In preforred embodiments,, the PRG4 has an average molar mass of bet wen 50 kDa and 400 Mt., and is .recomb_inealit PRG4, isol. t .ti .n.aturall -acct rraa l R4,, or a ftmctional fragment thereof.
l00161 In some crtrbe?clin3entsa the ophthalmic device of the current invention further comprise a lubricating composition associated with or otherwise rep .rsibly bound to surface active recet$or(s) (e.g., PRG4) or surface of the opthah c lens. In certain embodiments, the. lubricating composition described in the cu gent invention comprises a ('el forming agent or composition and/or a surfactant or surfactant composition, or a combination thereof, In one embodiment, the gel firming agent Or C-01,11POSA1011 Comprises hyaluronic acid or sodium hvaluronate. In mother eirmbodiment, the surfactant or surfactant composition comprises one or more surface active phospho.lids, such as l.:-co tlip ilmito\ llphosphdtid\ Iclboline, phosphatids kholine, } l c~ l l r:tici >letl rrrc iarr err c, and sphi_ngomye..hn. Described in certain embodiments of the present invention is the observation that the get forming or surfactant agent or composition associated with. the boundary lubricant molecules detach during a shear event, thereby itrev' itiElg the shear 1.5 stress from reaching the epithelial surface. Following the transient "'bearing, event, the gel farming and/or surfactant agent or corrmpo sition, alloyed to return to their undisturbed equilit rian r, rebind to the surface bound receptors and increase the probability of release from the receptor with increasing shear amplitude.- such that an one association is easily reversible.

100171 In farther or alternative embodiments, the surface hound receptors' comprise hya_lurotric acid. In this embodn:rment, the ophthalmic device of the current invention further comprises a lubricating agent. or composition assoc anted with, or otherwise reversibly bound to, the hi -aluronic acid. The lubricating composition in this embodiment, comprises a gel. forrtaing; composition comprising PIR04, and/or a surfactant composition comprising one or more surf ce. active phospholil ids, such as ] -a-dipalmitovipho lilt rtidilcholine, phosphaatadvlchol ne. I?lrocl?}r;rticÃo.l tlr;anolaurarac_ and sphingortm v el in [mutil In certain other embodiments. the surface hound receptors comprise DNS.
aptamers. I NA aptamers that may be utilized herein include those that recognize proteog lycans such as PRG I, liyaluronic acid, long chain sugars such as dextrans.
polyethylen e.. glycols, or other DNA constructs, and fi aturc tunable of nitv. through an {
iterative evolution are' selection, or through ratiometric des'jg -i against a semi con-.iplement .ne hybrid (a e,, a. purposefully a a.ismatched polvG-A-polyG
could act Is a surface bound receptor for a poly&T-poiy(strand. with shortening; lengths of polyG
incrzasin relative afrit-%).

S leadÃ9] in some embodiments, the sacrificial m chanism (e.g.. cornprising surface bound receptor(s)) is bound to the ophthalmic lens by rev'ensihle and/or irreversible interactions (e . .. covalent bonds, non-covalent interactions, or the like). In certain efl)hOdi:nlcnts, the present invention provides that the surface bound receptors are adhered to the ophthalmic leas surface by direct adsorption, hydrophobic Ionic. or covalent binding or by tinker 1.0 lac:ngistries se ected :fitarrr the group consisting of hoino- or hetero-bifunctional linkers, N _ h droxV sued imidyl esters, biotin, a.Vidin, streptavidirn, raaalcimide, thiol bonding.
Ãamiraes, hvdr zone . dendrisr er-s, and carbod imidc.s.

10tr20] Provided in certain embodiments herein is an ophthalmic device comprising an ophthalmic lens with an outer sute and an inner surface, PRG4, a P104 inducing 15 conmpound, or a combination thereof being associated with at least a portion of the outer or inner surf rcr in an amount et dive to provide ocular boundw, lubrication in an ocular environment of an individual wearing the ophthalmic lens. In some ,mbodiments, the PRG4 is associated in a manner so as to provide a sacrificial mecharrisni, as described herein, In certain embodiments. the PR04 is bound to the surface of the ophthalmic lens, 20 In some embodiments, a device described herein comprises a. lubricating composition disposed on the surface of the ophthalmic lens, the lubricating composition comprising () a gel-forming agent, a surfactant, or a combination thereof and (ii) optionally PRG4.

110021] In some embodiments, ltubricatir g, gel forming or surfactant composition further comprises one or more ophthalmically acceptable agents selected from the group 25 consisting of an ophth almically acceptable demulcent, ophthalmically acceptable excipient. ophthalmically acceptable astringent, ophthalmicalk acceptable wisoconstricto.r..
and ophthalmically acceptable emollient. Exemplary ophthal:naic ally acceptable demulcents contemplated in the present invention include, but are not limited to carbor.vrnethvlcellulose sodium (e.g., about 0.2 to 2.5'%) wv/v ), hydroxy:thyl cellulose (e.g., 30 about 0.2 to 2.5 are iv v). h ypromellose.. (e.g.. about 0.2 to 2.5% w/v ), mc;thvlcc;llralose about 0.2 to 2.3% w/v' , dextr an 70 c, ?., about 0.1 `o tt/v , gelatin k.g., about 0.01`~ar W/V , gl cerin (e.g., about 0.2 to 1%, N-vi`r.), polyethylene glycol 300 about 0.2 to 1% w /v )polyethylene glycol 400 (e.g.. about 0.2 to l?olvsorlar!: e 801.
about 0.2 to 1 propylene gKool (e.g., about 0.2 to 1% w- /Y), polyvinyl alcohol (e.g., about 0.1 to 4"=%
v /v), pov idone (e,g, about 0.1 to 2% 5 100221 Exemplary ophtl almically acceptable excip e is e.mohi nts contemplated in the present invention include, but am not .limited to, aanh, drous lanolin (e 8,, about I to 10", wA ), lanolin (e.g., about l to 10% wi / v), light :n1i:aneral oil i; .o._ =
about 50% car/v 3, mineral oil (e.g., = about 50".:o paraffin (e.g., - about 5"% ?:v/ v), petrolatum (..g , - about 100% ?\-Jv). white ointment about 100% wlrte pt.tras ttt}tn (e.g.. about 1.0 .100"t7 white wax (e.g.. ,= about 5% vv/v), yellow wax (e g., = about

5',",/o w,/V). An exe 3aplaa:r ' ophtllalmically acceptable astringent contemplated n the.
present invention inclcades, but is not limited to, zinc sulfate about 0-25% xenapla:ty ophthaln)icaily acceptable. vasoconstrictors contemplated in the present invention include, but are not limited to, ephedrine hydrochloride (e.g., about 0. 12w/v), naph<arzoline 1 5 hydrochloride about 0.01 to about 0_03% v v), ph enylephrine hvdmchlori e (e.g...
about W M to about 0.2% e : v), and, tetmha.d.rozoline hvdrochloride. about 0.01 to about 0.05"' xv/ `)-l(H)231 In some of these embodiments. the demulcents, exe l icnts, astringents, vasoconstrictors, emollients and ;lectrolstes provide a means to deliver the.
boundary 20 lubricant molecules in an ophll?.almicaly acceptable manner. phthalrnically acceptable compositions are suitable fbr topical application to the ocular Surface if the lack unacceptable eke toxicity, burning, itchiness, vtscersity, blurred vision.
etc. upon aappl ica:tion.

1(H)241 In certain embodiments, the gel forming or surfactant composition further 25 comprises other ophthalmic lens care compounds that may be suspended in a phosphate buffer .d saline or an osmotically balanced salt solution of tear electrolytes, including one or more of sodium chloride about 44 ,%, to 54%i mole fraction). potassium chloride about 8% to W%;, mole fraction), sodium bicarbonate }
4 about '~ ~:, to 18'X, :mole fraction), potassium bicarbonate f c.g., about 0% to 4% filole traction), caalciauzmj chloride 30 (e.g., about to 4'F~% mole faction). magnesium chloride. about 0% to 4%
mole fraction), trisodiaam citrate. (e.g. about 0% to 4% mole fraction), and hydrochloric acid Ãt (C .l .. about 0% to 000 mole fraction) or sodium hydroxide (e g.. about 0% to 20%) mole faction). In one embodiment. the carrier could be formulated to generate an aqueous electrolyte solution in the 150-200 mm mange.

10Ãr2 ,1 in certain embodiments, the ophthalmic lens care compounds are suspended in an ophthal.m-Ically acceptable balanced salt solution comprising at least three electrol.tiles, but not limited to, sodium chloride (NaC;I) 0.64%%%) potassium chloriÃde.
(KCI) including 0.07 r`)t') calcium chloride clihvdrata (C'a.C12-2:H20) 0,049',/i), aria :nesium chloride hexaliydrate (MgC12.6H20) 0.0-33'%, sodium acetate trtlaydrate (C2H Ni02-3U2CI=) 039%1- sodium citrate dehydrate (C0HSNa307.2H?0) 0.17%, sodium h`droxide andi or 1.0 by droc:h.loric acid (to adjust pH to approximately 7.5) with an osmolar-ity of a. proximately 300 rnOstns/L.

ittta261 in certain embodiments, the ophthalmic lens care compounds are suspended in an oplh.thalmically acceptable balanced salt solution, comprised of sodium (Na+) of approximately 128 mki, potassium (K-a-) of proxtmatele 2.4 mM, chloride CI-) of 15 approximately 11) mM, calcinrn (Car2-:) ofapproximately 0.4 n1M, magnesium (M82+) of approximately ("0 mM, HCO3- of approximately 5 aYrlt1 citrate of approximately I mrmM, phosphate of approximately 14 mM., acetate of approximately 15 .mM~ and sodium h~ dro idà ar:tcl: or' lr.~ dr'eachloric. acid (t-),adjtrst pH to app ximatelyy 7.5) swwitlr an osmolarity Of appaoximatcch.' 300 aniOsms L.

20 1002771 The present invention also provides an ophthalmic device comprising an ophthalmic lens With an outer sunccc and an :inner surface and an ocular houndaru lubricant composition disposed on at least a portion thoivof one. or more ocular boundary.
lubricating agent such as. by zsway of non-limiting example, of PRC 4, a PGR4 inducer, h aluronic acid, sodium hyaluron ate,, and/or a phospholipidte. ;., in an mount effective to 25 provide, alone or in combination wvith the surface bound receptor, ocular boundary lubrication in an individual wearing the ophthalmic lens). in some embodiments, a el-forming or surfactant composition utilized herein. comprises a gel forming agent and/or a surfactant, and an optional ocular boundary lubricating agent such as, ht=
Nvay of non-limiting example. PRC 4, a PGR4 inducer. hvaluronic acid, sod mn hyalu.mnate, and/or a 30 phospholipid (pre:icrrably present in an amount effective to provide, alone or in coraihination with the surface bound receptor, ocular houndar luhncation in an indin idual wearing the ophthalmic lens) Also provided in certain embodiments therein is a method for providing ocular boundary- lubrication to an individual in need thereof by applying to an eye of the individual an ophthalmic device of the present invention. In some instances, the invention method provides a sacrificial mechanism on the ophthalmic lens to mitigate Shear Stress, SO ,18 to treat ocular surface hvpcrosmoltiarits.

BRIEF DESCRIPTION OF THE DRAWINGS

100281 Figure I represents, feedback. loops within ocular surfaec bourrdaa :
lubrication.
10Ã}231 figure 2 illustrates PRG4 nlRNA expression in hwnan cornea! epithelial cells.
Human corneal epithelial cells were isolated from the coane:oscleral runs of male and 10 female donors. Amplified samples i wre screened for the presence of PRG4 products b using an Agilent 2100 Bioanalyzer. Vertical lanes contain: L, MW ladder 1. No template control., 2. Corneal tissue from a )3 year female; 4. Cultured conceal epithelial cells from a 70-year female; 6. Cultured corneal epithelial cells from a 53-tear male.

l.ta#a3O1 Figure 3 illustrates I'RG4 mR. A expression in human conjunctival epithelial cells.
1? 1-luman corneal epithelial cells were isolated from the corneosel ral .rims of male and female donors. Amplified samples were screened for the presence of PRG4 products by using atl arose gel electrophoresis. Vertical laces contaira. 1. MW ladder, 2.
No template control; 4-Harman female conjunctiva; 5 Humaan male co:nJuncti a.

l0 3t1 Figure 4 illustrates Ã'RG4 mR.N.A expression in human comeoscler al rim. tissue samples, 1- Human corneal epithelial cells were isolated from the corraeoscleral rims of male and fQmalc. donors, Amplified samples ti ere screened for the presence of products by using an Agileut 2100 Bioartaly zer. Vertical lanes contain: MW
ladder; I.
Human liver eDNA suandard; 2. Corneoseleral ring tissue from a 24-year female;
3.
Conacoscleral rim tissue, from a 5 1 year female; 4. :Human conjunctival epithelial cells.

lffl32l Figure S illustrates PRG4 n R.NA expression in human coaja-rctie.al impression cytology Sampies .Clrt.a jraraetival impression cytology mples w ;re. isolated tiom male and female donors. Amplified samples were screened for the presence of PRG4 products by using an Agil.eilt 2100 Bioaraalyzer. Vertical lanes contain: L. MW ladder; 1-9.

J. I.

Conjunctival impression cytology samples; 10. Repeat of human eoliluncfival epithelial cells (Lane 4 in Figure 3).

100331 Fi; Ure 6 Illustrates a friction test schematic. 'l-he corneal ocular surface (605) was fastened to the spherical end of an inert norr puineable semi-rigid rubber plug, cylinder (6031) (radius r:::6 mm). The plug cylinder (603) was attached to the rotational actuator of the mechanical testing machine (Bose ELF 3200) fonaningthe bottom articular surface. An an:nuhis (631.) (ou.ter.radius 3.2 mm, inner radius=1.5 mm) was punched from the eyelid (604)A he annulus (601) was attach d to the linear actuator coupled with an axial load (N) and torsion (=) Toad eells. Icrr nirrg the upper articulating surface.
Lubricant bath (602) was 1.0 formed by securing an inert tube around the plug cylinder (603). = rs the angular f equrency.

itrr)34j Fil iue 1 illustr rtes the rc duction of in vitro lid uc?rr c.a..kinetic friction with aaddition of PRG4 protein (h,rhric_in ).

100351 Fil ure illustrates the reduction of in vitro hd/cornea kinetic friction measured 1 14; minute after the addition offPRG4 protein (lubricin).

[031;l Figure 9 Illustrates the reduction of in wtro lid/c.orr:rea kinetic friction measured 5 minutes after the addition of PRG4 protein (lubrieiu), 100371 Figure lli illustrates the reduction of rrr vitro lid/cornea kinetic friction over time, following addition of PRG4 protein (luhric:un).

DETAILED DESCRIPTION OF THE INVENTION

]ttÃr3s1 Provided herein, are ophthalmic devices and methods for managing ocular boundary lubrication in association with ophthalmic lens wear. In certain embodiments, the invention modulates hyperosraa.olarity at the ocular surface c-ia a sacrificial mechanism to improve ocular boundary lubri ation. Prodded herein is an ophthalmic device 25 comprising an ophthalmic lens with all outer surface and an inner surface and a sacrificial mechanism disposed on at least a portion thereof in an amount efft:cti v . to provide ocular boundary lubrication in an ocular environment in an individual wearing; the ophthalmic lens.

1D0391 Though not wishing to he hound by theoretical mechanisms of action, as shown in Figure I, increased shear stress leads to tear film instabilityy, evaporative tear loss..
hyperosmolarity, changes in swelling pressure and a feedback elevation in shear stress. In some instances, increased shear stress promotes inflammation. androgen deficiency and decreased expression of prot;o lycans. In certain instances increased shear stress and its seÃluelaac, may... over time. lead to a loss of boundary lubrication at the ocular surface. A
deficiency in ocular lubrication and symptoms associated therewith can be determined by any suitable raethcd. In some insmnces, a deficiency in ocular .hub ncation a a_nd symptoms associated therewith is do. fined eitla.er claalitaitav el. (e.g., a feefing) of low- lubrication, dry ey . discomfort, etc) or quanta ativelz (c;.sg- measured through mechanical, hiochen.arl, electrcal, optical or other methods of tlra,antitaati e assays).

10040l in certain instances, and as prodded herein. PR64 protein plays a critical role in the eve as a boundary lubricant. In some instances, this secreted gl :coprÃot ::in protects the ocular surface to protect the conica and conjunctiva against significant shear fc,)rcos generated during, an e did blink, contact lens wear, and any other undesirable ocular boundary lubrication caused b y chronic inflammation and hypcrosmo!aarity that result from dry eve disease, androgen deficiency, estrogen replacement therapy, compromised gear film. allergy, aanirn.g, ocular surface diseas, and increased protease levels in the tear film and at the ocular suurface. Given the relationship between ostriotic pressure afid tile electromechanical interactions s oth.in charged molecules, the present invention provides, in sonic embodiments, a pharmaceutical composition for managing a deficiency in ocular lubrication b modulating hvperosmol:aritv or osmolarit at the ocular surtf'iacee via interrupting the feedback mechanisms that pre vent secreted components from reducing friction coefficients and mitigating shear stress.

lI004ll The present invention features a sacrificial mechanism for ocular boundary lubrication in association with ophthalmic fens wear, whereby sunface hound receptors reversibly bind to a lubricating composition. in sonic emix)(fiments, the lubricating composition comprises one or more ge.l forming acid/or surfactant agents or compositions.
In some instances, the get forming or surfactant composition detach during a shear event, thereby preventing the shear stress from reaching (or reducing the, shear stress reaching) the epithelial surface. in certain enibodinaents, follow-ing the transient shearing event, the gel Ãornuing and surfactant composition, allowed to return to their undisturbed equilibrium, rebind to the surface bound receptors. In some. embodiments, the entire composition can detach during shear. In certain instances, that the thermodynamics of this, equilibrium increase the probability of release from the receptor with increasing shear amplitude. but such that any one association is easily reversible.

1(H)421 ilh.erefo:re, the current invention generally features a leg., approach to ocular lubrication in the presence of an ophthalmic. lens. In particular, provided herein is a mechanism or process that relates to the use of a sacrificial mechanism to reduce friction at the interface between the, ocular- surface and. an ophthalmic lens, including t uundary 1.0 lubricant molecules such. as PRO4. l %al.uroraic acrid, sodium ivaiuronate, and phospholipids.

10Ha431 As used herein, an "ophthalmic lens" refers to lense which are placed:
in intimate contact with the eve or tear fluid., such as contact lenses for vision correction (e.g., spherical, tonic, bifocal), contact lenses for modification of eye color, ophthalmic drag 15 delivery devices, ocular tissue protective devices (e.g., ophthalmic healing promoting lenses), and the like.. A prof rred ophthalmic l--is is an extended-wear contact lens, especially extended-wear contact lenses for vision correction. with oxygen transmissibility or penncabrht~. ion pennestabilit . gas permeability, and other desirable transmissibility or pemreabi.lit and fetatures_ As used herein, an "ocular environment refers to ocular fluids ?0 (e.g., tear fluid) and ocular tissue. (e.g_, the cornea.; which may come, into intimate. contact with, a contact leas used liar vision correction, drug delisverz, 'ound healing, eye color modification, or other ophthalmic applications.

l'tHt441 As used. herein, an "outer surf ace" of Cin ophthalmic dears r6er, to the anterior surface of the lens which faces av,rw,' from the eve during wear. Thc outer surface, which is 25 typic a.llt~ substantially convex, may also be referred to as the front curve of the lens. The `inner surface" of a lens, as used herein, refers to the posterior surface of the lens which faces towards the eve during wear. The inner sur -ace. which is t-%ypicaliy substantially concave, may also be re:ferred to as the base curÃre. of the lens.

l(H)451 In one embodiment of the current. invention, the sacrificial mechanism comprises a 3 0 pluralit : of surface bound receptor comprising PRG4, and a lubricating composition reversibly bound to PRG4, wherein the lubricating composition comprises . gel `forming co..mpositio.n comprising hyaluronic acid or sodium h-yaluronate, or a starlactant composition comprising one or iricire surface active phospholipids, such as, dipaiiiiiitoylphoslph.atitl.y lwlholinte, phosphatidvlchoiine, 1~l~os lr>
t:i~lo l ilr>tnÃ~I~a i rc, and S sphingomyel ra. In this embodiment, PRG4 is allowed to interact with endogenous proteins and proteogit Bans within the tear film, and the exogenously supplied hyaluronie acid at -id/or phospholipids to establish a sacrificial mechanism to paecent protein or lipid adsorption, at the lens surface, reduce dry spots on the lens, and reduce. the friction bet -een the lens and the ocular surface, In f .,et another feature of this embodnraaent, the hyahirimc acid and/or phospholip.ids are replenished in the form of a topical artificial tear drop, reaching solution, contact leas cleaning products, an overnight incubation, or other contact lens care products.

l00461 As used herein, the terms "PRG4", " PRG4 protein' or "`pr.)teoglycan 47" and -lubricin" are used interchanmably..PRC 4 is used herein also to encompass the tetra 1.5 megalcaroc1e; stimulating factor (M.SF), that has been accepted for the ÃiCLJltGGNC,,I-HUGO Human Gene, Nomenclature data base, and superficial zone protein (SZP). The PRG4 or lubricin protein as used herein refers to any isolated or purified native or recombinant lub.ric:in proteins, homolo s., famcfional fragments or motifs, isofo ms, and/or mutants thereof. In certain embodiments. the isolated. or purified PRG4 protein comprise s an amino acid sequence for a human native.. or recombinant.
lubricin protein. In other embodiments, the isolated. or purified PRG4 protein comprises an amino acid sequence encoded by Cplg-/gene axons that encode the full length P,64 protein or isofonns' primary structures. The proteoglycann 4 (pro4) game contains 12) ee:wons. The PRG4 protein used herein can comprise an ammo acid sequenc encoded by prg4gen exons l -12, more preferably. axons 6-1.2, and most preferably. exons 9-12.

100471 As used heroin, the.l'"I G4 protein includes any MCA proteins now kno tin, or later described. In certain embodiments, a preferred PRG4 protein amino acid sequence is provided in SEQ ID NO.1. The P .C 4 protein shares the primary amino acid structure, of any known PR64 proteins or isofonns with at least 60"r%% homology. prcferably 754) homology, more. preferably 85% 90"s-, 95%. c3ft%, 97%, 98%. 9 % or more homology, In certain embodiments, a preferred PRG4 protein has an average molar mass of between 50 kDa and 400 kDa, comprising one or more biological active portions of the PR
,4 protein, tar faanction a:l ira nacnts saach as a laibric atin ira nacnt, or ca laorarc5loÃ* thereof 100481 In yet another embodiments #tanctioa3Kal ira aaae.ntsa multimers (e.g..
d_irners, trimer'ss tetramers. etc'). homology or orthologs of PRC3 act. as the surih receptor and/or gel 5 forming constructs in d w sacrificial mechanism.. Functional fiagments and homology of PRG4 include those with a fever repeats within the central mucin like KEPA.PTI'-:rep,'at domain, glycosylated and non-glycosylated forms of the protein, splice variants, r .combinant forty s. and the like. A lubricating fragment of PRG4 exhibits at least 20',!,4.1 30% 40%, 50%, 60%, 70%, SO%, 90%, or 95% of the ophthalmic lubricating efftct of 1.0 human PRG 1, as measured qu'tl:itatiseh, nacc.lrm.icall e optiuall\:, electrically- or by biochemical assay, 1(#)49] As used herein, the PRO 4 protein comprises a biological active portion of the protein. As used herein, a ""biologically active portion" of the PRÃ 4 protein includes a f notional fragment of a protein comprising to mo acid sequences sufficiently 15 homologous to, or derived. from, the amino acid sequence of the protein, which Includes thwer amino acids than the, f=ull len4Yth protein, and exhibits at least one activity of the. Ball-length protein. Typically a biologically ac:tix-e portion comprises a functional domain or motif with iat Ie ast one acts its of the l rotein, biolo ;k ill actr e.
portion of a. prof in can be a poly'peptide which is, for example, 10, 25, 50. 100. 200, or more amino acids in length. In one embodiment, a biologically active portion of the PRG4 protein can be used as a therapeutic agent alone or iii combination with other therapeutic agent's for treating undesirable or decreased ocular bomidar lubrication.

l0 $0] The nucleic acid. and amino acid sequences of several native and re;.coml_in ant PRG4 or Iubricin proteins, and characterization of the PROS- proteins and various isoforms are disclosed in, for instance, US. P;;at.e;nt.'Nos. S 32o,' 5 $ 6,433,142:
T030,2123: T361. 738 to `ruiner ct al.. and U.S Patent Nos. 6.743.774 and 6.960.:62 to Jay et al..
U.S.
Publication No, 20070 19 1 268 to Flannery et al. also discloses recombinant PRG4 or lubricin molecules useful in the present invention.

l(H)5i.] Methods for isolation, purif cation, and recombinant expression of a PRG4 Protein are well known in the art. in certain embodiments, the method starts with cloning and isolating mRNA and cDNA encoding PRG4 proteins or isoforms using standard molecular biology techniques, such as PCR or RT-PCR. The isolated cDNA encoding the PRG4 Protein. or isofi rraa is then cloned into an expression -vector. and further transformed and expressed in a host cell for producing recombinant PRG4 protei s.

100,521 As used herein. srecoa abinant" refers to a polvnucleotidc synthesized or otherwise manipulated in vifm "recombinant polyllucleotide"'), to methods of using recombinant polynucleotides to produce gene products in cells or other biological systenms, or to a polypeptide: (`recombirnant protein') encoded by a recorrmbinarn poly uc eotide.
``Reconabi.nant" also encompasses the ligation of nucleic acids hating various coding 1.0 regions or domains or promoter sequences from different sources into an expression cassette or vector for expression of, e.g.. inducible or constitutive expression of a fusion protein comprising an active domain of the PRG4 gene and a nucleic acid sequence axrt7lrlifled using a primer of the invention, 100531 In certain embodiments. the PR64 protein encoding nucleic acid may contain one or more mutations, deletions, or insertions, In such embodiments. the PRG4 Protein encoding nucleic acid i4 at least 60firõ homology, pr ter;:bly 75% homology, more preferably 85%, 940%, 9 5%, 96`%/%), 97%. 98%, 99'i %. or more ho.mmolo y7 to a wild type PRG$ protein, encoding nucleic acid.

10541 As used herein, the term cDNA a' includes D: IA that is eomptenientarz to naRN
molecules present in a cell or organism mRNA that can be convened into c.D -A
with. an enzyme such. as reverse tr rriscriptase, In certain embodime ts, the cDNA
encoding PRG4 protein is isolated from PRG4 n RNA expressed in hwnan corneal or conjunctival r pitlielial cells using an RT-PCR method well know-vii in the art.

It a 5l is a ed herein, the terms ' pola.iairclcrrtirl~ ." -)-micicic acid/nucleotide,," and "oli ;onueleotide" are used interchangeably, and include polymeric forms of nucleotides of any length, either deoxvrihonuc.leotides or ribonarcleotides,s or analogs thereof Polvnucl :otides m l'i have anti' three-dimensional structure, and may pertom am functions, know a or unkrno q n The following are non-limiting examples of polynucletiotides: a gene or gene. fragment- exons, introns, messenger RNA (rnRNA), transfer RNA, ribosomal 34:i RNA. riboz apes. DNA, cDNA, wenorlliic DNA. recombinant pol nucleotides, branched pol,-mucieotides, plasmids, sectors, isolated DNA of any sequence, isolated RNA of any seq,uerace, nucleic acid probes,, and primers. Polvnucleotides may he naturally-occurring, synthetic, recombinant c_or aiiy combination thereof, l(msol A polyuauc.leotidee may comprise: modified tatacl.c tides, such as mcthylated nucleoticlcs and nucleotide analogs. If present, modifications to the nucleotide structure may he imparted before or after <issemlaly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polymicleotide may he further, modified after polymerization, such as by conjugation with a. labeling component. The term also includes both double- and siramle-stranded, molecules, Unless otherwise specified 1.0 or required, any embodiment of this invention. that is a poivnucl;;otidc encompasses both the double-str aaided f ?raai and each of mc-o eomplcuietatarr saat~ le-st.rtaatcl d toraias known or predicted to make up the double.=stranded form.

[M) ,571 As used herein, the term "polvnucleotide sequence," is the alphabetical representation) of a pol nucleotide molecule. A polvnnucleotide is composed of a specific sequence of truer nucleotide bases: adenine (A); cytosine (C f> guaaninc (U);
thNiniiue 1,.1,3.
and uracil (U} is place of thymine when the polyntic.leotide is RNA, instead of DNA. This alphabetical rep csentation can he inputted into databases in a computer and used for hioinformatics applications such as, for example, functional genomanics and homology searà bin .

100581 As used herein, the term "isolated poly'niuucleoti.der'c.DNA." includes polvnucleotidc molecules which. are separated from other polynucleotide molecules which are present in the natural source of the polyrmeleotide. For example., with regard to genomic DNA, the term 'iso_ilated".' includes pc_olyna.ccleotid.e molecules which are separated from the chromosome w.itli which the ge.nÃ?mic DNA is naturally associated Pzefe ably, an 'isolated-` polvnucl ootide is free of sequences which naturally flank the polyrtucieotidc (i.e.. sequences located at the 5' and 3' ends of the pol 'nucleotide of interest) in the genomic DNA of the organism from which the polynuc.leotide is derived. For example iii various embodiments, the isolated polvnnacieotide molecule encoding the PRG4 protein used in the invention can contain less than about 5 kb. 4 kb, 3 kb, 2 kb, I
kb, 0,5 kb or 0.1 kb of nucleotide sequences i. hich naturally flank the polyriucle.otide molecule in genamic D`+i. of the cell from which the pol nucteoti.dc is derived. Moreover, an "isolated"

J, t3 polt'nucleotidc molecule, such as a cD1" A molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized.
l0Ã 591 As used herein, a "gene" includes a polvnnuclcotidc containing at least one open reading frame that is capable of encoding a particular polypeptide or protein after being transcribed and translated . An -%.- of the pol :nucleotide. sequences described. herein may also be used to identify larger fragments or bill-ten tlr coding sequences of the gone %vith e l ia.la the-. are associated. Methods of isolating larger fragment sequences are known to those of skill i.a the art. As used herein, a "native or raatatr;allti<trccurrirr " polynucleotide 1.0 molecule includes- for example, an R.NA or DNA molecule having a nucleotide sequence that occurs in nature (e.g.. encodes a natural protein).

ioo of As ,used herein, the tem "polypeptide" or "protein" is interchangeable, and includes a compound of two or more subunit amino acids, amino acid analogs, or peptidomimetics. The subunits may be linked by peptide bonds. in another embodiment,.
the subunit may be linked by other bonds, e.g?.. ester, etlaea, etc. As used "herein, the terra amino acid" includes either natural and/or unnatural or synthetic amino acids.
including glycine and both the D or L optical isomers., and amino acid analogs and peptidommietics.
A peptide of three or more amino acids is commonly referred to as an oligol eptide?.
Peptide chains of trreater than three or more amino acids are referred to as a potpeptide or a protein.

100611 In certain embodiments, the PR G4 protein used herein refers to PRG4 proteins or Various homol.ogs or is4brnas thet :s_of, that are naturally or rccosnbinantly expressed in humans or other host cells. As used heroin, "express' or "expression" includes the process by which polrnucleotides are transcribed into RNA rands=or translated into po ypeptides. If '5 the polsnuclcotide is derived from g normtaic DNA, expression rna include splicing of the RNA if an appropriate e:ukar otic host is selected. Regulatory elements required for expression include promoter sequences to bind RNA 11olyMaerase and transcription initiation sequences for ribosome binding. For example, a bacterial expression vector includes a promoter such as the lac. promoter- and for transcription initiation the Shiner Dali arno sequence and the start codon AUG. Similarly, a eul aac"otic expression Vector includes a heterologous or homologous promoter :ftor RNA polyme:rtase IL a downstream lq laolvaden'laa.tion signal, the start codon AUG, and a termination codon for detachment of the ribosome. Such vectors can be obtained commercially or assembled by the sequences described. in methods well known in the art, for example, the methods described below for constructing vectors in general. As used herein, the ten-a3 "vector-7 includes a self-replicaating nucleic acid molecule. that transfers an inserted pohnaaclcotide into andlor between host cells. The. terra is intended to include vectors that function primarily for msertton of a nucleic acid molect-d into a cell, replication sectors that function primarily for the replication of nucleic; acid and expression vectors that l unction for transcription and/or era nslation. of the DNA or RNÃA. Also intended are vectors that provide more than one of the above function, i0621 As used herein. at `-host cell" is intended to include any individual cell or cell culture which can be. or has been, a recipient ::lor vectors or for the incorporation of exogenous polynucleotides and/or polypeptides. It :is also intended to include progeny of a single cell. The progeny may not necessaril be completely identical fin morphology o s, in , enonaic or total :DNA comple:nne:nt) to the original parent cell due to natural, accidental.
or deliberate mutatiorn. The cells may, be prokar-ti,otic or eukar 'otic, and include but are not limited to bacteria] cells, yeast cells, insect cells.- animal cells, and mammalian cells, including but not limited to mucirne, raat. simian or human cells. As used herein, a host.
cell" also includes genetically modified cells. The term "gca tically modified cells"
includes cells containing and/or expressing a foreign or exog--enoius gene or polyntic] cotide sequence which in turn modifies the genotype or phenotype of the cell or its progeny.
"Genet.icaalk> modified" also includes a cell containing or expressing a gene:
or poivntic] ec tid : sequence which has been introduced. into the cell. For example, in this embodiment, a genetically modified cell has had intr-3c uced a gone which genre is also 2: endogenous to the cell. The term "genetic ally modified" also .includes any addition, deletion., or disruption to a cell's endogenous nucleotides. As used herein, a "host cell" can be any cells that express a human PRG4 protein.

100631 As used herein, "lacarnolous" are defined herein as two nucleic acids or peptides that have similar, or substantially identical, nucleic acids or amino acid sequernces, respectively. The term "homolog" further encompasses nucleic acid molecules that differ from. one of the nucleotide sequences due to degeneracy of the genetic code and thus encodes the same amino acid sequences, fn one of the preferred embodiments, homologs include allelic variants, orÃhologs,, paralogs, rtà o sts, and antagonists of nucleic acids encoding the P R64 protein SEQ ID NO: 1t.

10 641 641 As used herein, the term ` ortliologs " reth s to two nucleic acids fro .m different 5 species. but that have evolved from a common ancestral gene by spec ation.
Normally, orthologs encode peptides having the same or similar functions. In particular, orthologs of the invention Will generally e .h:ibit at least 80-85y'.=x;, more preferably 85-90% or 90-95%, and most preferably 9?% 96"%, 9M, 98 .tt. or even 99`..%) identity. or 10(X,, sequence.
identity, with all or part of the amino acid sequence of any known PRG-4 proteins (e.g..
.1.0 SEQ ID NO: ]), isofornisa or analogs thereof, and will exhibit a function similar to these peptides. As also used therein, the term "paralogs" refers to two nucleic acids that are related by duplication w rthirt a genuine. Par-alogs usually have different ('unctions, but these functions may he related.

10001 To deteta:ine the percent sequence identity of ttcwo amino acid sequences, the 15 sequences are aligned fbr optimal comparisom purposes (e.g.. gaps can be introduced in the sequence of one poll peptide for optimal alignment with the other polypep ide or nucleic acid). The amino acid residues at corresponding amino acid positions are then compared.
When, a position in one sequence is, occupied by' the same amino acid residue as the corresponding position in the other sequence, then the mol culees are identical at that 20 position. The same type of comparison can be made between two nucleic acid sequences.
Ilse percent sequence identittiy between the two sequences Is, a f6action of the number of identical position., shared by the sequences (, e., percent sequence identity . numbers of identical positions/total. numbers of positions x 100). Preferably, the isolated amino acid honlologs included in the present invention are at least about 50-60%, preferably at least about 60-70%, and more preferably at least about 7(# 7 "'is 75-80%. 80-85%. or-90-95 's'%, and most preferably at least about 96%, 97%, 38 '`rx 99%, or more identical to an entire.. amino acid sequence ofany~ known PRG4 protein (e.g., SEQ III NO, 1).

100661 In certain embodiments- an isolated nucleic acid homolog encoding the PRG l protein comprises a nracleotide sequence which is at least about 40-6W,,,'), pr e.f ra:bly at least.
about 60-70%, more preferably at least about 70-75%r;a. 75-8M 80-85%7 85-90%.
or 90-and oven. more prefer rbly at least -about 95 %%a 96'%, r 7e1` . 98%'.'%,_ 99'%, or more identical to a nucleotide sequence encoding amino acid sequences of such PRG4 Protein SEQ ID NO:1).

100671 The detemiination of the percent sequence idcntitw between two nucleic acid or peptide sequences is well known in the art. For insstan ce_ the Vector NTI 6.0 (PC software package if.n[c?r la , Bethesda., MD to determine the percent sequence identity between two nucleic acid or peptide sequences can he used, In this method, a gap opening penalty of 15 and a gap extension penalty of 6.66 are used for determining; the.
percent identity of two nucleic acids. A gal) opening penalty of 10 and a gap extension penalty of 0, l are used for deter pining the percent Identity of two polypepticles. All other p rrameters are set at the 1.0 default settila gs. For purposes of a multiple alignment ('laastal r algorithm) the gap opening penalty is 10, and the gap extension penalty is 0.05 with blosumti2 matrix: It is to be understood that f(-or the purposes of determining sequence identity when compatin a 13NA sequence to an R.N.A. sequence, a tlwn idine nucleotide Is equivalent to a aaracil nucleotide.

1tl0681 furthermore, the PRG4 protein used herein includes PRG4 protein encoded, by a polynu :l :.otide that hybridizes to the polynudeotidc encoding, PRG4 protein rider stringent conditions. AS Used herein.. "hybridization" includes a reaction in which one or more pr_r1 nucleotides react to form a complex that is stabilized via hydrogen bonding between the bases of the nucleotide residues. The hydrogen bonding may occur by ?t:) Watson-Crick lase paining, I Ãc?c, teirt l?.finding, or in ans other sequence-specific rnamler.
The complex may comprise two strands f >rming a duplex sti-tiettire. three or more strands forming a multi-stranded coraaplex, a. single self-hybridizing strand, or any combination of these. A. hybridization ruction may constitme a step in a more extensive process, such as the initiation of a PCR reaction, or the e=nzw n=tatic cleavage of a polw nucleotide by a rabozvme..

100691 : t bridizati.on roactions c .i be performed miter different stringent conditions. "rile present invention includes polynuc.leotides capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and. most preferably higlily stringent conditions, to polti nucleotides encoding PR6I protein described he ei.n. As used herein, the term. "stringent conditions' ref =rs to hybrid.izatiort overflight at 60" in W x x Den.h art's solution. 6xSSC, 0.5% S.DS. and 100 mg/ml denatured salmon sperm DNA.

Blots are i ashed sequentially at 62='C fir 30 minutes each time in 3 SSÃ'/0.l% SOS.
folloNved by IxSSC/0.1 ":%, SIBS, and finally 0`i xSSC (O f % SDS..As also used herein, in certain embodiments, the phrase "stringent conditions" refers to h ybridizaation in a 6xSSC
solution at 63' 'C, la other embodiments h Yhly stringent conditions" refer to hybridization oycnri ht at 65 C in IOxD:nhart's solution, 6xSSC, 0.5% SDS and l.00 mg/ml denatured salmon sperm DNA. Blots are washed sequentially at 65"C for 30 minutes each time in 3tSSC/0.l% SDS. Followed by ixSSC/0.lc'i, SDS, and f rally 0. IxSSC/0.l % SDS. l lelhods for nucleic acid by bridizatiÃons are well known in the art.
Accordi.n ly. the PRG4 pr teins encoded by nucleic acids used herein include nucleic acid haying at least 60`% homology, preferably 75% homology, more preferably 85%, more ' 99g'. =0 .
~JI'C.'LI~1 ~' most 1?#'G~~%Ii~~.?l~4' 96U. iQ1110 C.~g' ut' to a polynucleotide sequence that encodes a human PRG4 protein SEQ ID NO:!) or a specific iso.llo rm. or horno log thereof.

l.tHt70l Moreover, the PRG4 proteins used herein can also be chimeric protein or fusion protein. As used herein, a "chimeric; protein" ear Tfia:siora protein"
Comprises a first poltpcptit=lc op uttivO linked to a second pol .laeptide. C:hirneric proteins mrr optionally comprise a third, fourth or fifth or other polypeptide operatively linked to a. first or second polype.ptidc_ Chimeric proteins may comprise two or n oree different polypcptidcs.
Chia acne Proteins may Comprise multiple copies of the same laolypeptide.
Chime ric proteins may also comprise one or more mutations in one or more of the polypeptides.
Methods for making chimeric proteins are yell known in the art. In certain embodiments of the Present Invention, the chimeric protein is a chimera. o.f PR 4 protein with other PR.G4 protein isoforms.

100711 As used herein, an "isolated." or "pun died." protein, laolNlnu.cleotide or molecule.
means removed from the en vironraaent in which they naturally occur, or substantially free of cellular material, such as other contaminating proteins from the cell or tissue. source from wvhich the protein poly nucleotide: or molecule is derived., or substantially free from chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular matori.aal4' includes preparations separated from cellular components of the cells from which it is .isolated or .recombinantly produced or synthesized. in certain embodiments., the language '-substantially free of cellular nimater al"

includes preparations of a PRG4 protein having less than aibotif 30% (byd weight) of other proteins (also referred to herein as a ` c,ont as n.inat:ing rotei.n"), more prefe nabls less than about ]IM, still more preferably less than about lt)".~i>, and most pre.f :rablv less than about 5% of other proteins. When the protein or pol r ueleotide is recoin inantll produced, it is also preferably substantially free of culture l edium, i, f., culture medium represents less than about 20%. more preferably less than about 1.0%, and most prefer-ably less than about 5 %% oftl e Volume of the preparation die protein of in era st.

107,21 In certain em bodirnents of the, currant Invention... the surface bound receptors comprise hy-alrrrorric acid. In this embodiment, the lubricating composition reversibly 1.0 bound to the. by aaluronie acid, wherein the lubricating colnpos:ition comprises a gel for :rrirag composition comprising PR. &4 and a surthctant coi pos to comprising one or more surface active phospholipid.s, mel rcling but not limited to, Ã_-a-di ahrlito lph.os h;atidy lclrolilre. lalros laat d lelat l r lit slat atid~ l tl ~ alcrlarltine, and sphingonr elfin.

15 IO0711 The present invention also provides an ophthalmic device comprising an ophthalmic tens with an outer surface and a inner surface and an ocular boundary lubricant composition disposed on at least a portion thereof one or more ocular boundary lubricant molecules selected from the group consisring of PRG4, a PGR4 inducer.
hv>ahironic aid..
sodium lip al.arrx mate, and a plaoslphol:ilsida in an amoar:nt effective to provide ocular 20 bounds Imbrication in a patient v-veaarirag the ophthalmic lens.

1'(0741 As used herein, as "PRQ4 inducing compound" or "PR G4 inducerr' refers to a compound that increases the bioconcentration of.PRG4, e.g., .a compound that is capable of upregulating PRG4 exprc sion, promoting the biosynthesis of PRG4..
inhibiting' degradation of .PRG4. or the like, includi.n but not limited to, an androgen or androgen 2S analogue, selective androgen receptor rr rodulator. selective estrogen.
receptor modulator, estrogen antagonist, aronlaataase inhibitor, aratiprot.ase, proinflammatorv.
eyytoki.ne ,antagonist (e.-g. selected from the group consisting of nt:i-T.NN.l~c antibody, soluble "TNFa receptor, and I:L-I receptor antagonist,), cs tokine release inhibitor, a arriiinfammatory cvtokine (e.g. TGT`- 3). anti.inflammatory agent (e.g. cycloslx)rin A, omega 3 and 6 fatty acids), NF-K B inhibitor, or proteaaso.me inhibitor, and pharmaceutically acceptable carriers for topical use, l' 075] In yet another embodiment, the androgen or androgen analogue as selected from the group consisting of 17cc-aaaetla 117 ~la cirri ? o 'cr~aarafirc+stara~~~cne derivative, a 3 it.r g n su sti ated an rog:~n, a testosterone derivative, is a 4,_5 ~~-clih drote..stt~sreron dens ative, a i9-aortestosterone derivative, . 1.7 I ro ri < -~t durst ttr derivative containing a ring .A unsaturation,, and a stmctural subclass of androgens comprising androgenic compounds with unusual structural features.

10076] In another preferred embodiment, the selective androgen receptor modulators (S A.RI Is) are selected from a group consisting of aryl-propionainide. S_3-(4-sace:ttlami:no-pheno~`)-2-hydrox -2-meth l~1 -Ã4-nitre>-3-tr f uotometh 1-leper i)-propio.narnide J.Src4], or S-3-(4-fl u:oroplh.eaox) 2-hvdro>.1 2-metlhvl-N--l4-tm.itro- 3õ
t.ritltacaronictla i-l laen~ l)rl rc l io ranud S-i ]), hicsdic hydantoin, cluinoli.n ;, and t tral ~ drocitFita.oli.ta.e aà al goes that have in-vivo atidroge.nic and am olic activity of a non-1 5 steroidal ligarad. fbr the androgen receptor.

100771 In vet another preferred embodiment. the selective estrogen receptor modulators (SERMs) are non-steroidal ligands of the estrogen receptor that are capable of inducing as number of conformational changes in the receptor and eliciting a variety of distinct biologic profiles. Preferably, the SERMs are those that prevent estrogen--induced inflammation in ocular surface tissues. In cermin preferred embodiments, the estrogen antagonists are, steroidal or non-steroidal compounds .nldepeaarlea t of receptor affinities.

it ,781 Other molecules may also be used as surface bound receptors within the sacrificial mechanism Of the Current Invention. For instance, DNA sequesiecs recognizing fgel for acing or surfactant compositions (e.g, DNA aptamers), bvould serve as the surface bound receptaor. These aptamers could recogYriize proteoglycans such as JITW14, hyaluronic acid., long drain sugars such as dex-.trans_ polyethylene glycols, or other DNA coast nets.
The surface bound DN.A could feature tunable affinity through an iterative cc olutionaar selection, car through ratiometrre design against a semi-complementary hybrid (i_e., a purposefully mismatched polvG-,.-polvG could act as a surface bound receptor for a polyG-T-polvG strand, with shortening lengÃhs ofpoly increasing relative afnitv.].

1007`3] In certain embodiments, th ; surface bound receptors are.. adhered to the ophthalmic lens surface by direct adso.rption,, hydrophobic ionic, or covalent binding or by linker chemistries selected from the group consisting of homo- or heter,)-bifunctional linkers, N_ h\droxti succinimid 'l e:stcla, biotin, avidin, sta'eptavidin, mal,,unide.
tlnol boudia ;, 5 amines, hvdrazones, dendrim.ersa and carbodiinaides, Methods for disposing, adhering '7 coating. or attaching the. su)-facie bound receptors, or other desirable molecules, the aaistries, monomers. oligont.e:rs, and polymers, to the ophthalmic lens are well known in the art.

100801 In certain embodiments, the present invention provides that the gel t firming or 1.0 surfactant composition of the current invention further comprises one or more then apeuticeally effective amount or concentration of oplhathalmically compatible and/or acceptable agents selected from the group consisting of an ophthahnicalls acceptable dc:aagaalcent, excipient, astringent, vasoconstrictor, and emollient. As used herein, the term "ophthalmically compatible" refers to a material or surface of a material which may he in intimate contact with the ocular environment fbr an extended period of time without significantly damaging the ocular environment and without significant user discon:at_irt.
Thus, an ophthalmically compatible contact lens will not produce significant corneal swelling, will adequately move on the eye with blinking to promote adequate tear exchange, will not have substantial amounts of lipid adsorption. and will not cause substantial wearer discomfort during the prescribed period of wear.

100811 As used herein, the term efteaeti e concentration or amount" or "flier apeu:ti.call1 eth ctive concentration or amount- is intended to mean a nontoxic but suit cie:nt concentration or amount to provide the desired therapeutic effects, The concentration or amount that is eMcti} e. will var from sub ect to sail ject, depending on the atge and general condition of the individual, the particular agents, and the like.
'Ibus, it is not alp. ays possible to specify an exact: effective concentration or amount, flowwwe er, an alalaao priate effective concentration or amount in any individual case may be deter gained by one of ordinary skill in the, art using routine experimentation.
Furthermore, the exact effective concentration or amount of the boundary lubricant molecules used herein and other therapeutic agent incorporated thereinto or dosage form of the present invention is not critical, so long as the concentration is Within a range sufficient to permit ready application of the solution or formulation so as to deliver an amount of the boundary lubricant molecules and other active agents that is within a therapeutically effective range.
100821 In certain embodiments., the pharmaceutically efe.ctive concentration of PRG4 protein is in a range of 10-l0,000 .tgf L. preferably 50-5.00 nn õ{inL, and more preferably li-(1 300 ms-,','M l ..: s used laert in, the of lrtlz~ilinieall ' acceptable agents comprising the oplithaximieail acceptable demulcents, csc.il icaats. astringents.
vasoconstrictors, and emollients that are fully defined in the Code of Federal l egulations 21C.FR349.

l00831 -l:n certain embodiments, the lubricating composition described .herein comprises or the aforementioned ophthalrnicalh acceptable agents are or can be combined with one or more of carbwoucthvlccllulose sodium (e.g.. about 0.2 to about 15% hvdroxy :.thvl cellulose (e.g.- about 0.2 to about 1i%, ~~,'~ .Marc>mellosc (e.,g.. about 0.2 to about 2.5%
w/v), methvicellulose (e.g. about 0.2 to about 2.5%~, wfv), dextran 70 (e about 0.1%
gelatin (c'-g., about 0.01 % glycerin (e.g., about 0.2 to about 1% /v).
polyethylene glycol 300 (e.g. . about 0.2 to about 1% w/ ), poh ethvlene glycol 400 (e. g..
about 0.2 to about I% s , polysorbate 90 (e.g, about 0.2 to about l `# v propylene ,glycol (e.g. about 0.2 to about polyvinyl alcohol e.g.. {about 0.1 to about 4iN) vvv). povidone (e.g., about O.J. to about 22'N% w/), zinc sulfite (e.g., about 0.25 'x% w/ V), anhydrous lanolin (e. ;., about I to about 10`?i, 1-v/0, lanolin (e.g.,. about 1 to about l0%
w/v). light: mineral oil (e.g., . about 50% wi'v), mineral oil (e.g.. = about 5t]rNO wt'/v).

paraffin (e.g., ' about 5% 11'/v), petrolatum (e.g.. = about 10(lz.'%~.
4'e''"~'.}. white ointment (e. y...
= about WWW:'V). white Petrolatum (e.g'., = about 100% v,, N), white wax (e.g, - about 5% w/v), yellow ryas (e.' g- = about ti.phedrine hydrochloride (e.g., about 0.123"='i, v /v), naphazoline hydrochloride (e.g., about 0.01 to about 0.03"%, /v), hetaslephrine hydrochloride (cg., about 0.tl8 to about 0.2 ~% ww:? v), and te.trali ydrozolin hydrochloride (e. g., about 0.01 to about 0.05% w/x. ). In certain 'instances. percent amounts utilized herein are percent amounts by k >eight.

100841 In f rther embodiments, the therapeutically effective concentration of hyalurwowc acid or sodium by aluronaw is in the range of 10-100,000 tag/in l ., preferably 500-5,000 mg/n~iL, and the therapeutically elf ..ctive concentration of the surface active phospholipids is in the range of 10-1.0,000 ugr'mL, sud-t sutftu. _ active phospholipids iacludc, but are not limited to, :Ã,H= il? t:ln ittp lphosp tid l .holm (DPPC.`}, p.hosphatid ylcholine. (P+ ..
plt 4l?l?rt[id i~.[l?r tal.trt? na. (PE) and sphingomyelin (p), or other neural and polar lipids, 100851 The lubricating composition as disclosed herein may further comprises one or more phaarrnaaceutically acceptable carriers or vehicles comprising any acceptable raateriaals, and/or any one or more additives know in the art As used herein, the term "carriers" or vehi.cle" refer to carrier materials suitable for topical dmg administration.
Carriers and vehicles useful herein include any such materials known in the art, which are nontoxic and do not interact with other components of the composition in a deleterious manner. Various additives, known to those skilled in the aW, may be. Included in the 1.0 composition. For example, solvents, including relatively small amounts of alcohol', may be used to sol ubil.ize certain drug subst ices. Other optional additives include opacifiers, antioxidants, fragrance, colt?rant. gelling agents, thickening gents, stabilizers, surfactants, and the like. Other agents may also be added, such as antimicrobial a ents, to prevent spoilage upon story / ., to inhibit growth of microbes such as yeasts and molds.
15 Suitable antimicrobial agents are typically selected from the group consisting of the methyl and pr-opyl esters of p-hydr_pxvbenzoic acid (i.e., methyl and propyl paraberr)), sodium benzoate, sc?rbic acid. imidureaa, and combinations thereof. Permeation enhancers and/or irrita tioai~raaiti raaiira~õ additives nm also be imftidcd in the pharmaceutical Composition of the present Invention.

20 [(H)861 In certain :embodiments, [ht pharmaceutically acceptable carrier compris'e's a phosphate Wooed. saline or an osmotically balanced salt solution of tear electroly=tes, including one or more of sodium chloride in about 44% to about 54`,',(, mole fraction, potassium chloride in about 8% to about .14' mole fraction, sodium bicarbonate in about 8% f+
l~.o to about t}rE, mole fraction, potassium scar oa1;a[e in about t%, to about `f,:'/u anv 25 fraction, calcium chloride in about l#% to ibotit =I`.'=) mole fraction.
alaaagne.starn chloyride in about 0% to about 4% mole fraction. trisodium citrate in about Wlllt to about 4% mole fraction, and hydrochloric acid in about 0% to about 20% mole. fraction or sodiam hydroxide in, about 0%, to about 20% mole traction. In c :rtarn embodimhats, the pharmaceutical carrier can be Formulated to g nerate an aqueous el :c'trolyte solution in 30 about 1-0-200 rM .t ange, Wier suitable formulations, such as ointments, creams, gels, pastes, and the like, suitable for topical administration, are also contemplated in the present invention, Ill certain embodiments, electrolytes provide proper osmotic balance when combined with PRG4 to make a solution ophthaimicall ' tcceptahle.

l+aÃr8 71 The present ins ent:iora further provides a method for providing ocular boundam lubrication to an individual in need thereof co xrpri.sirna applying to an ev of the individual an ophthalmic device comprising an ophthalmic lens N-with an outer surface and a inner surface and an ocular boundary .lubricant composition disposed on at least a portion thereof one or more ocular boundar lubricant molecules selected from the group consisting of P.Q4, a PG4 inducer, hvaluronic acid. sodium hyaluronate., and a phosphollpidt in an a n oua.nt efIbctive to provide ocular boundary lubrication i -t an I.0 individual svear'ing? the ophthahnic lens. In one eamfbodiment, the invention method. is used for treating ocular aurf ac hyperosmolarits in the individual who wear the ophthalmic lens. The invention method provides a sacrificial mechanism on. the ophthalmic lens to mitigate shear stress; as discussed above.

100881 'Thron.ghout this application, various publications are ref :renced.
The disclosures of all of these publications and those references cited within those publications in their entireties are. her bye incorporated by r ference into this application iii order to more fully describe the state of the art to which this invention pertaiats.

1.(H)891 It should also be a nlerstood that the foregoing relates to preferred embodiments of the present Invention and that numerous changes may be made therein w -ithout departing from the scope of the invention. The invention is further illustrated by the following examples which are not to he construed in any way as imposing limitations upon the scope thereof. On the contrary, it is to be clearly understood that resort.
mat be had to various other embodiments, mod. fications, and equivalents thereoÃ: which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the present. invention and/or the scope of the appended claims.

100x901 Other features and advantages of the invention will be apparent f:ix-?m the folio vine d; scription of the, pretene.d embodiment thr. reof and from the claims, `These and rnarav other variations and embodiments of the anventio:n .c ill be apparent to one of skill in the art upoon a review of the appends description and, examples.

EXAMPLE'S
EXAMPLE I

PRG4 mRNA Expression in Human Corne=al and Co ajunctival Epithelial Cells 0091] Human cuan.eaal epithehal cells r.twer isolated from the corneoscleral firms, of male and female donors. Cells were processed either directly (ra = 8). or first cultured is phenol red-free keratinoCvIe serum f ee media (aa = 2). Bulbar conja.ua_C[ivae (rr =
2). conjunctival impression cytology samples (a W 9), immort hz d human conjunciic,azl epithe=lial cells after culture (n 1). NOD mouse lacrimal lands (a 5 adult mice/sox', 10 gltands/samplc). and BALB/t mouse meihoram au glands (n 7 adult mice/sex, glands from 28 lids/sample) were obtained durim,, surgical procedures. 'These samples were processed for the analysis of PRG4 mRNA by using primar'il , RT-PCR (n = 1S human, all mouse) and Affti metrix GeneChips (n = 4 human corneas). 'Tlhe PRG4 primers for PCR
spanned over I .kbp of intaonn sequences- to order to suppress amplification. of contaminating chromosomal DNA (Table 1). Amplified samples were screened for the presence of 14; products by using ag arose gel electaclat e resin and. an dent 7100 ioaEaaah zer. To confirm the identity of anaplicons. PCR products from cornea samples (n 2), conjunctival epithelial cells (n W 1) and a human liver standard: (a = 1) were sequenced with a 3100 Genetic Analyzer at the Massachuse=tts Eye and Far Infinnany. DNA
Sequencing Center for Vision Research (Boston. MA) w id resulting data were analyzed with BAST searches of GenBank data bases.

T~ l~le l . C3ligoea.aacleotacle primers designed for U- PCR analysis of11RG4 rn.R.NA
Species Orientation 1`taac leotide sequence (5. - 3) Exons Amplicon Size (bp) Human Sense GA GCAG+ TT ACCC(AAA (SEQ 11) NO: 2) 9-.122 526 Antisense C:ACiACTI- CGATA.AGGTTC. GCC (SEQ ID NO:3) 1092] It was demonstrated that PRG4 mRNA is present n all human corneal and conjunctival, e pitlheli i cell and impression Cytology samples. '11e identity of PRG4 PCR
products was confirmed by D NA sequence analysis (Table 2). The results show that PRG4 is transcribed in human conical and conjunctival epithelial cells.

Table. .2 Identification of amplicon quences from human cornea, conjunctival and liver samples Sequencing Aligned Base Pairs `total Base Pairs BLASTh Search Direction To Human PRG4 from Amplicop Identtir' 1-luman Liver Standard A l-onvaad 4535 500 Harman PRG4 A Reverse. 488 491 Human PRG4 10 B Forward 496 499 1-hum m PRG4 B Reverse 498 500 Human PRG4 Human Cornea t24 year Ãaltl :feanaa:le A Forward 497 499 Human PRGI4 15 A Reverse 490 492 Human PRG4 B Forward 500 504 Hà inaan PRG4 B Reverse 498 501 Hu m a_n 11RG4 1-luman Cornea (51 year old female) 20 A Ron,,.,ard 498 499 Human PRG4 Revcrst:. 474 489 Hunan PRG4 B Forward 496 498 Human PRG4 B Reverse 490 491 Human PRG4 25 Han= C annjun.cÃivaal EPitlic al Cells A Forward 496 499 Human PRG4 * Reverse 490 4922 Human PRG4 B Forward 495 499 Human PRG4 B Reverse 474 491 Human P1RG4 Two ditl:cren{ saamp:les (A&- B) of each pz l.,rrattczzt '+ker SequCn ced in forward w id reverse di eetioaas. the human corm !,t s<tanrt?les were el?ialxeliaal cells fro-.r11 the cà awl)"c:ieraal rims of feInnle dotnors. '111e ,Zen accession xtauttlaer fear human PR04 is NM O0-W)7.

PXAMPLP.2 's Reduction of friction in vvitro with the addittion of PRG4 (lub icianl loO93I An in vitro friction test with clinically relevant interfaces, such as an Ocular surf c -c'slid and ocular surface-contact lens interface is described. las lta~>,. Clinically relevant methods capable of quaantit ati elti assessing the lubricating ability of artificial tears are currentl. lacking. Friction tests z.vith synthetic (e.g. latex and glass) or nova-ocular `nat:ive' surf aces (C-g- umbilical cord veica segments) may facilitate some, but likely not all 01 the molecular interactions that occur during indeed_ the tele aance of data, obtained w 'ith non-tisstie interfaces as unclear.

3l.
1D0941 An annulus-on-disk rotational test configuration has been shown to he ideal for studying boundaryy lubrication at an articular cartilage-cartilage interface.
A boundary mode of lubrication is indicated, by kinetic friction being is ariant with factors that influence formation of a fluid film, Including sliding ve.locit and axial .load. This is because surface-to-surface contact is occurring, and surface bound molecules contribute to lubrication (by decreasing friction and wear). Boundary lubrication has been discovered to be a critical and operative mechanism at the ocular surface. like it is at tlhw articular cartilage surface. Thercfoae, the in v/fro friction test past, ious.ly developed and characterized to study boundary .lubrication at an articular cartilage-cartilage interlace was lÃD modified for the study of ocular sure cc-cvc lid and ocular surf,.ace-contact lens interfaces.
lows], To determine the test conditions in which boundary lubrication is dominant at the ocular surface - sYelid and ocular surace-contact lens iuterfac .s._ the dependence of frictional properties on axial load and sliding velocity, was examined. Normal fresh hum-cm ocular surfaces (:rescctecl corneas with 3Ism of sclera) were obtained from t ho lions Eve 15 Barak of Alberta. The resected corneas were stored in Optisol-GS it 4`}C
and used t Within 2 i ee:l s. Eyelids (age 60-8() years old) were obtained f'roru the University of Calgary Body Donation Pro gram within 1-3 days after death and used i:naraaeahatelm or stored at 20='C in saline for at most 2 weeks until use. Comparative lubricants consisted of lens Plus Sterile Saline Solution (Advanced Medical Optics) as a negative control" S Stan "
Lubricant Eye 20 Drops (Alcon Laboratories), Refresh 'Fears Lubricant Eve Drops (Allergan), AgquifY""~
Long Lasting Comfort Drops (C1.B.A. Vision) and B.li:nk fear` Lubricant Eve Drops (Advanced Medical Optics) as test lubricants.

100961 The friction -test schematic is shown in Figure 6. The conical ocular surface (605) was fastened to the spherical end of an inert non-permeable semi-rigid rubber plug 25 cylinder (60) (radius r:::6rnm) by applying super gluee to the sclenu. '11-us plug, Cylinder (603) was attached to the rotational actuator of the mechanical testing machine (BoseELF
2200) thus, forming the bottom articular surface. An annulus (601) (outer radiu =3.2mm.
inner radius 1.5mm) was punched from the eyelid (604), and -: as attached to the linear actuator coupled with an axial load. (N) and, torsion (=) load cell, thus forming the upper 30 articulating urfac:. Lubricant )lath 602 was formed by- securing, an inert tube around the plug cylinder ('60' ).

[W971 Samples were first tested in saline, then in one of the three () test lubricants. The lubric ai:tt bath was filled v,-,ith -O.3 anl,, and the articulating surfaces allowed to equilibrate with, the lubricant. The sample sur-ftices were slowly (0.05minIs) brought into contact and compressed until the, spherical plug flattened out and the entire annular eyelid star lade was in contact with. the cornea (605). The resulting normal stress (calculated from axial load as, in units of MPa, as N/(= ,r utas r snn e ~ can be varied by using dif treat stif less rubber Plugs to tni.naic physiological stresses W a. The test ,,,Q clucnce was initiated by preconditioning the sample by rotating +4 revolutions (rev) and reset with -4 revolutions at a physiologically relevant effective linear sliding velocity, veff = 30 mm/s (where veff Reif; = is the angular frequency- and lsefi::2.4mm is the effective radius calculated by integrating the Shear stress distribution over the annular contact area).
Sataaples were then tested by rotating .4 revolutions., immediately followed by 4 reset revolutions at veff W
30. 1.0, 1. 0.3 and then `0 rnmis_ with a. dwell time ot~. 12 second between each revolution.
The test sequence was then be repeated in the opposite direction of rotation.

1n0981 To evaluate the lubrication properties of the ocular surface, two friction coefficients l=) of the :form =:::=/(R,,r ) ) where is torque. R;:rra is efh ctive rad.iaus, and N is axial load, described above. A static friction coefficient, which reflects the resistance to the onset ofmoti.on. ',tarn,- was calculated as the peak value of -,just after (within -10') the start of rotation, An average kinetic .friction co, l.f icient, v 'hich reflects the resistance. to steady state motion, was calculated from = aver-aged during the third and fourth co:taaplete test re v oluti.on. Both = oa f , and = sn.t s were averaged for the .,nil revolutions in each test to account .f-tar- potential directional effects on =
measurements, Data was collected at a. frequency of 20 Hz.

lta099l The results of lubricin (P'RG ) added to the corneal surface at a concentration in the range of 100-300 ug'mL. are. shown trt Figure 7, Lubriciu h aci a friction lowering effect.
at the eyelid inteahace. both in terms of kinetic, and static friction, at all velocities. At a concentration 1/10th of that of physiological hyaluronic acid, lubricin was similar to Blink" Tears Lubricant Eyre Drops, which contains hvtaluronic acid, In combination, the two lubricants are better than either alone.

lootottl Figure 8 demonstrates the reduction of in vi 3ro comeailid kinetic friction measured dining the first minute after the addition of l.ubrie.in as compared to ;Aq iih ' eye drops.

Lubricants were thoroughly washed from the ocular surface. using saline be wecn tests. A
synergistic of ct (reduced over either alone) was evident when Aquii~" (with h aluronic acid) was combined with lubaricin. '1 he saline repeat was loswwer than the original saline control. This showed a. retention o luh sc n's of ct even after washing with saline, suggesting that the molecules were binding to the ocular surface- and that lubricin demonstrated superior retention time as compared to sodium by aluzonat e alone.

1t mu1 Figure 9 demonstrates the reduction of in vitro cor eai.Iid kinetic friction measured during the 5th minute after the addition of Iubricin. as compared to Aquila "
eve drops..
~F>
synergistic ef'ftct (reduced } zxx t~ over Mier alone) Nvas evident t hen Aquil ~ ith 1.0 hvaluronic acid) was combined with .lubricin. he friction coefficient of :Aquatt` had returned to statistical. equivalence to saline after- 5 ia.unutes, w. hereas lub-iciaa remains lower, as did t e combination of lubricirn and hyalurouic acid.

1M1.02"1 Figure -l tl shows the meduetion of kinetic friction coefficient over time, following addition of lubricin. Again, the continual reduction suggested binding to the ocular 15 surface.

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SEQUENCE LIST

SEQ ID \O: I

N- A:WK'1LPIYLLLLLSVFVI QVSSQDLSSCA RCGE: YSR . 'IC. C.DDYNCQHY'M
EC( PDFK'R\=CTAELSCKGRCFE_ :FERGRE(DC DAQ(KK\~ DK(C:PDYESR AEVI-IN
PT SPPSSKKl'\PI'PSC r'-SQ'TIKS'1"FKRSI'KPPNKKK'IK.K'\'IESEEI`I'EEHS
VSL.\QL.SSS
SSSSSSSSSTIR-KIKSSK\S A.A\RELQK.K.L KN, KD\KKNRTKKKPTPKPPVVDEAGS
GLD' 'GDFK''TTPDTSTTOI-INKVSTSPKI I."':'tKPINPRPSLPP SDTSKETSLT\'NI :
TT <'ETKMTTNI TSTDGKEKTTSAKETQSIEI TSAI DI:APTSK `1.AKPTPKAt: T
ET'KG.P. I_.'ET PK.1:P rP'I'I PKf- P AS-I."T P.K.FITI. P"t I-'II S: P I-I P .F
P,A.P'FFIT.i KSA.P"T 1 PKF P
AP'T 1 I I .T::P'Ak.I''1`I'T3I f .I'.r .P'I'-1"I :.I I':'e1' I-I'I'KS:.t '.1'1'F'I<;I .I : T''I'-1'PKI PAT'T'-I'T}I I:,T'. P.I, TPKLPTPTTPKEPAPTTKEPAPTTPkLPx\PT:'PKKPAPTTPKEP:r PTTPKEPPTTT'K
EPSPTTPKEPAPTTTKS:I:P`I TKEPAPT"(TKS. PTTPKEPSPTTTKEPAPITPK P \PT
TPK:kPAPTTPK .P_1P'ITPI EP LP1T1'KKPAPTTPKEPAPTTPKLT.k.PTTPKKLTPTTP
FK1a. PTTPFKPAPTTPFICI.NPTTPFFPTPTTPFFPAPTTPk:AA, P\TPKFP?APTTPKF.
I:- PAPTTPKFP. PTTPK.V'TAPTTPKCITAP'TTLI .FP.APT rPKKPA.PI .FI-:kPTTTKTP'TSTT
CD {P:APT`I'PKG'I.APTTPK.Pd>I T'PKEP P'FI'PK(G 1'r'\PFPLKF.PAP'FI'1PK_KPAPKI L
AI''I IT'. . 1"T.'S'.Y"1.'SI)KPAP'Y'I.'PI 1 Tx- I'V...PK PAPTI'I I KP P
Tl'I)E"1..PT'P'.P"I"SEVS'i.'P
`P ITKLP'f'I'IP- KSI) IFS'I'PELS EP'I'I)K Li NSKEPOVI' 1 1"P 'P'1'AA"P'1 IIEM'1 1"l' AK1 KITERDLRTTPETTTA.LPK\ITKLT:ITTTEI, .TTESKITATTTQVTSTTTQDTTPFKIT
TLKTTTL;\.PPC\' TTTKKTITTTLIM\KPEET:tKPKDR. TNSK_ .TTPKPQKPTKr PKK
PTSTKKPKT\I.PRVRKPKTTP'TPRKMTS'TMIPFI.\PTSRI. EAM1.QTTTRPNQTP SK
I_:\`I'\'\PKSE713ACsCi.A.T::G.I--' IP.HM1.I_:IR.PH\'EN2P.b\' I'.PD:
SID\'I,PR\'P:NQCG.IIINPM,L:SD
E" T'FDIC:' GK.P\`DGP.'.IIT..RMG'I.I..\;: F.RGHYE'\ {MI.SPF
S.PPSP;A.RRP'T'F\'\ `ccI'PSPII)'-I'\
1='I'RC\CE(jK'I'I FFKI)SQ\' :.'R1 T-N_DIKDAG ' PKPIFKG1 GGLT'GQPVAALS'I:AK' `K
5 'N\\ 'PLSV`I`r:PkRCct3SIQQ\`I 'KQEPVQKCPC5RRPAL\. 'P\ VGETTQ\ RRRRFE.RAI
GPSQTI-ITIRI.QYSPr' RLA\ QDKG.IVLHNEVK' SII.\\,'RC LPI 'L'VTSAISLPNIRKPDG
YD'S YAFSK.DQY NIDVPSRTARAITTRSGQTLSI :VWYNCP

SEQ ID NO 2: G ATGCAGGGT:ACCC CAAA (human. sense ) :.30 SEQ ID NO.3: C AGGACTTTCIGAT:A.AGGTCTGCC (Iicman. anit se ns._)

Claims (23)

What is claimed is:

1. An ophthalmic device comprising an ophthalmic lens with an outer surface and an inner surface and a sacrificial mechanism disposed on at least a portion of the outer or inner surface in amount effective to provide ocular boundary lubrication in an ocular environment of an individual wearing the ophthalmic lens.

2. The ophthalmic device of claim 1, wherein the, sacrificial mechanism comprises a plurality of surface bound receptors.

3. The ophthalmic device of claim 2, wherein the surface bound receptors comprise surface bound PGR4.

4. The ophthalmic device of claim 3, wherein the device further comprises a lubricating composition associated with the surface bound PGR4.

5. The ophthalmic device of claim 4, wherein the lubricating composition comprises a gel forming agent, a surfactant, or a combination thereof.

6. The ophthalmic device of claim 5, wherein the lubricating composition comprises at least one gel forming agent, the at least one gel forming agent comprising an effective amount of hyaluronic acid, sodium hyaluronate, or a combination thereof.

7. The ophthalmic device of claim 5, wherein the surfactant comprises an effective amount of one or more surface active phospholipids.

8. The ophthalmic device of claim 7, wherein the surfactant active phospholipid is selected from the consisting of 1-.alpha.-dipalimitoylphosphatidylcholine.
phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin.

9. The ophthalmic device of claim 2, wherein the surface bound receptors comprise hyaluronic acid, and wherein the device further comprises a lubricating composition reversibly associated with the hyaluronic acid, wherein the lubricating composition is selected from the group consisting of a gel forming composition comprising PGR4 and a surfactant composition comprising one or more surface active phospholipids.

10. The ophthalmic device of claim 9, wherein the surfactant active phospholipid is selected from the group consisting of 1-.alpha.-dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin.

11. The ophthalmic device of claim 2, wherein the surface bound receptors comprise DNA aptamers.

.12. The ophthalmic device of claim 1, effective to inhibit protein or lipid adsorption on the outer surface, inner surface, or both surfaces of the ophthalmic lens.

13. The ophthalmic device of claim 3, wherein the PRG4 has in average molar mass of between 50 kDa and 400 kDa, and comprises recombinant PGR4, isolated naturally occurring PGR4, or a functional fragment thereof.

14. The ophthalmic device of claim 2, wherein the surface bound receptors are adhered to the ophthalmic lens surface by direct adsorption, hydrophobic ionic, or covalent binding or by linker chemistries selected from the group consisting of homo- or hetero-bifunctional linkers, N-hydroxy succinimidyl esters, biotin, avidin, streptavidin, maleimide, thiol bonding, amines, hydrazones, dendrimers, and carbodiimides.

15. The ophthalmic device of claim 5, wherein the lubricating composition further comprises one or more ophthalmically acceptable agents selected from the group consisting of an ophthalmically acceptable demulcent, an ophthalmically acceptable excipient, an ophthalmically acceptable astringent, an ophthalmically acceptable vasoconstrictor, an ophthalmically acceptable emollient, and tear electrolytes.

16. An ophthalmic device comprising an ophthalmic lens with an outer surface and an inner surface and an ocular boundary lubricant composition disposed on at least a portion thereof one or more ocular boundary lubricant molecules selected from the group consisting of PRG4, a PGR4 inducer, hyaluronic acid, sodium hyaluronate, and one or more phospholipids, in an amount effective to provide ocular boundary lubrication in an ocular environment of an individual wearing the ophthalmic lens.

17. A method for providing ocular boundary lubrication to an individual in need thereof who wears an ophthalmic lens, comprising applying to an eye of the individual an ophthalmic device comprising an ophthalmic lens with an outer surface and an inner surface and an ocular boundary lubricant composition disposed on at least a portion thereof one or more ocular boundary lubricant molecules selected from the group consisting of PRG4, a PGR4 inducer, hyaluronic acid, sodium hyaluronate, and a phospholipid, in an amount effective to provide ocular boundary lubrication in an ocular environment in the individual.

18. The method of claim 17, wherein the ophthalmic device is selected from any device of claims 1-15.

19. The method of claim 18, wherein the individual is in need of treatment for ocular surface hyperosmolarity.

20. The method of claim 18, wherein the method provides a sacrificial mechanism on the ophthalmic lens to mitigate shear stress.

21. An ophthalmic device comprising an ophthalmic lens with an outer surface and an inner surface; and PRG4, a PRG4 inducing compound, or a combination thereof associated with at least a portion of the outer or inner surface in an amount effective to provide ocular boundary lubrication in an ocular environment of an individual wearing the ophthalmic lens.

22. The ophthalmic device of claim 21, comprising PRG4 bound to the surface of the ophthalmic lens.

23. The ophthalmic device of claim 21, further comprising a lubricating composition disposed on the surface of the ophthalmic lens, the lubricating composition comprising a gel-forming agent, a surfactant, or a combination thereof.