CA2722944A1 - Therapeutic modulation of ocular surface lubrication - Google Patents

Abstract

Provided herein are ophthalmically acceptable pharmaceutical compositions comprising a PRG4 inducing compound i.alpha. combination with PRG4 (including a lubricant fragments, homologs, or isoforms thereof), and methods of using the same The PRG4 inducing compound in the pharmaceutical composition of the present invention upregulates PRG4 expression and localization in the ocular surface for efficient surface boundary lubrication. In some instances, pharmaceutical compositions described herein are utilized for treating ophthalmic conditions, e.g., ocular boundary deficiency and symptoms associated therewith.

Description

2 PCT/US2009/043015 Therapeutic Modulation of Ocular Stirface Lubrication CROSS-Rl~FER NC,.E TO RELATED APPLICATIONS

ltatrtl This patent application claims priority benefit of U.S. Provisional Application No.
61/05 1,1.1.2 filed May 7, '200K. 1, hich is incorporated hcnnin by reference.

FIELD OF TH E INVENTION

1'021 The present invention relate to the management of ocular lubrication. In particular, the present invention relates to pharmaceutical compositions, and method of use thereof for treatintu diseases associated with compromised lubrication at the corneal and conjunctival surfaces.

1.0 BACKGROUND

itrta31 The proteog?-IN,can 4 :rFg4t gene encodes f r highly glyeosylated proteins termed meLY~ I,~arvoc te. stiratarlatirr factor (MS' ), lubricin, and superficial zone protein (SZP) {.t}.
These molecules are eollcetisely re{ rred to as PRG4 or PR64 proteins. PRG4 is present in svnnovial fluid and at the surface of svnovicraaa. (2), tendon (3). and meniscus (4) and is suspected as being an important co nponent for healthy s ynovial joints. See_ e.g.. (5), (6) [(H)41 In tissues such <a_s sv. note ial joints, plat sicochentic.al modes of lubrication have been classified as fluid film or boundar, The operative, lubrication modes depend on the uornmal, and tangential forces on the articulating tissues, on the relative rate of tangential motion between these surfaces, and on the time history of both loading and motion.. The friction e r llicient, p. provides a quantitative measur., and is d :fned as the ratio of tan- ntial friction force to the normal force. One type of fluid-mediated lubrication mode is hydrostatic. At the onset of loading and typically for a prolonged duration, the in ;rstitial fluid within cartilage becomes pressurized., due to the hiphasic nature of the tissue; fluid ma v also be forced into the asperities between articular surfaces through a stie . p i n g mechanism. Pressurized interstitial fluid and trapped lubricant pools m a therefore contribute sigmficantl4 to the bearing of normal load with little resistance to shear force . facilitating a very. low p. Also, at the onset of loading, and/or .motion. squeen film, hydrodynamic, and elastol drodynamic types of fluid film lubrication occur. with pressurization, motion, and deformation tÃtinu to drive viscous lubricant from mid/or through the gap between two surfaces in relative motion.

1000) The relevant extent to which fluid pressun film versus boundary:
lubrication occurs classically depend: on a number of factors (13). When lubricant .film can flow.ww between the conforming sliding surfaces, which can deform elastically, i lastol o dro i ar e lubrication occurs. Pressures surface rorr d ness, and relative sliding; velocity deter acne when full fluid lubrication begins to break down and the lubrication enters new re;imes. As velocity decreases further, lubricant films adherent to the articulating surf-ac :s begin to contribute and a mixed regime of lubrication occurs. If the velocity decreases even further and only, 1.0 an ultra-thin lubricant layer composed of a few molecules remain, boundary labricatioll occurs. A boundary mode of lubrication is therefore. indicated by a friction coefficient (ratio of the measured frictional force between two contacting surfaces in relative motion to the applied normal force) during steady sliding being invariant with factors that influence :formation of a .fluid :fl.m, such as relative sliding velocity and axial load (14).
For articular cartilage, it has been concluded boundÃa.r-sy lubrication is certain to occur..
although complemented 1 fluid pressurization and other mechanisms t / _18).

10061 In boundary- lubrication, load is supported by surface-to-surface contact, and the ass_rciated.1ri.ctiotial properties are d.c:.term ned by lubricant surface molecules. "Thais node has been proposed to be important bectause the opposing cartilage layers mike contact over -10% of the total area, and this may be where most of tic, friction occurs (1.9), Furthermore, itla increasing loading time and dissipation of hydrostatic pressure, lubricant-coated surfaces bear an increasingly higher portion of the load relative to pressurind fluid, and consequently, this mode. can become increasingly dominant (13, 2 V).
Boundary lubrication, in essence, ma itigates stick:-slip (1$), and 'is therefore manrfi:st as decreased resistance both to steady motion and the start-up of motion. 'i'he latter situation is relevant to load hetaring articulating surfaces after prolonged compressive loading (e.g.
.sitting or standing in vivo) (21), Typicaal wear patters of cartilage surfaces 2') also suggest that boundary lubrication of articular cartilage is critical to the protection and maintenance of the articular surf. acre structure. 30 l007l With increasing loading time and dissipation of hydrostatic pressure, lubricant-coasted surfaces bear an increrasi 3gl higher portion of the load relative to pressurized fluid, and consequently, li can become increasingly dominated by this mode of lubricationn. A
boundary mode of lubrication is indicated by values of li during stead", sliding being invariant w itli fhctors that influence formation of a fluid filial, such as relative. sliding elocitv and axial load, Boundary lubrication, in essence, mitigates stick s.lili. and is therefore manifest as decreased resistance both to steady .motion and the start-up of motion.

l.tH)Sl The precise mechanisms of boundary lubrication at biological interfaces are currenÃl unknown. Hoawevor, proteoglvcan 4 (PRG4) may liia\ a critical role as a boundaiN~ lubricant in articulating _joirnts. This secreted glycciprotcin is thought to protect 1.0 cartilaginous surfaces against frictional forces, cell adhesion and protein deposition.
Various nrati ve. and recombinant lubricin proteins and isoforms have been isolated and tharaicterize.d. For i.usumcc:, +.S #`at.ut Nos. 5326,i581: 6.4331_142: 7 .0 022? and 7,361,7 38 disclose a family of human :mcgaka. oc yte stimulation factors (MSFs) and pharmaceutical Compositions containing one or more such MSFs for treating disease states 15 or disorders, such as a deficiency of platelets, l S. Patent Nos. 6.960.562 and 6343,774 also disclose a l.aibricaatin polvpeptide. tribornectiri, comprising a substantially pare fragment of .SF, and methods of lubricating joints by administering tribonectin systemically or directly to tissues.

1(H) )l A challenge to boundary lubrication is the presencee of inf atai.mation in surrounding 20 tissues, as well as increased protease levels in the stinoviail fluid. Loss of the boundaar.
lubricating ability of synovial :fluid af=ter ii jar y is associated with damage to the articular cartilage matrix. This can be attributed to inflammatory processes resulting from the injury particularly in the carl\ phases. Another challenge to boundary lubrication is a sex steroid imbalance, especially in arthritic disorders such as rheumatoid arthritis. Sex 25 steroids are involved in the pathogenesis and regulation of inflanrrmation ill rhei.uinatoid arthritis, a disease characterized by chronic inflammatory synovitis.
Androgens suppress, whereas estrogens promote, inflammatory processes. Consequently, the relative levels of androgens and estrogens in the svaaovial environment are extreinely important in determining the progression of inflammation (7. 8, _,). Various androgen compounds 30 reduce the magnitude of lymphocyte innf#ltration in lacrimal tissue Sec, e.g,, U.S. Patent Nos. 5.620,921: 5.688.765: 5,620.921: aalnd 6.107.2891 SUNIAM-ARY OF THE INVENTION

10OiO1 =The present Invention provi.desa in various embodiments, pharmaceutical corampositions, and methods of use thereof fir managing ocular .lubric<ation, including the therapeutic replenishment and enrichment of boundary lubricant molecules at the ocular surface.

[fruit} The present invention provides d -w discovers that PRG4 mRNA is expressed in human corneal and con uncti v al. epithelial cells.- as well as in mouse lacrimal and aa7eibc rniarn `lands, indicating that .PRG4 protein is presented in these tissues on the ocular surf ace, In addition, PR64 l rot aaa se .s to Prot ct the come a w id conjttncti a against significant shear forces generated during an eyelid blink, contact: lens wear, and other undesirable condi.tions. The impact of the tear film, including the impact of inflammation, pro] nlammaator v cytokines, sex steroid imbalance and proteases on the composition and function of the films., provide a course of therapy for ocular issues which promotes boundary Imbrication.

I0Ã}12] In certain embodiments. the present invention provides a pharmaceutical composition suÃtable for topical application to an ocular sai.rtce comprising a ther~~alaerit.ic aJlt effective concentration of a PRG4 inducing compound. In certain embodiments., the pharmaceutical composition further comprises an ophthalmicalla acceptable agent that increases the residence time of the PRG4 inducing compound on the ocular surface. In further or alternative embodiments, a pharmaceutical composition described herein comprises a PRG4 inducing compound in combination with a therape u:ticall .,- effective concentration of PRG4. 'I"he PRG4 inducing compounds encompassed. in the present invention Include. but are not limited to, an androgen, 4 as androgen analogue, a selective androgen receptor modulator, a selective estrogen receptor modulator, an estrogen antagonist., an au'c martat c inhibitor, an aarrtiprotcase, a proiariiam.matory cytokine antagonist, a c vtok a ce release inhibitor, an anti Ãrafl,armaamator c\ tokinc, an :aantiinfa.nimaators= agent, r N1^ -r;-I3 inhibitor- and a.
proteatiome inhibitor.
Described in certain instances of the present invention is the observation that T'R:Q4 expression in corneal and conjunctival epithelial cells is upregulated by the R:G-4 inducing compounds, as discussed above, thus. providing synergistic protection of cornea and contuntti'a against significant shear forces is ith PRC 4.

1D01.3] In certain embodiments, the a androgen analogues include, but are..
not limited to 17cxrnacth l-1'? -htciro -? a a- e _aandresstaanr? ne derivative, a nitrogen-substituted androgen, a testosterone derivative, a 4,5e.- iihvdreoti stost route derivative, a 19-nortestosterone derivative, a 1? -hvdre~~ - -araelrc sta e derivative conta fining a ring A
nrrsatait atieti, or a structural subclass of andro -ens composing androgenic compounds with atnta.sti.rtl. structurtal teaatu.res.

1(H)141 In some embodiments, the selective androgen receptor modulators (SARN-is) include_ but ar not limited to, aryl-propionanricie compound, such as S-3-(4-acety]aarai)'lo-~..hetio~~
1.0 or S-?-(4_fluorop.lietnox -f--2-h,droxv-2-methyl-NÃ-(4--nitro- -trifluoronmithyl-phetxvi)-propiagnara3idc [S-1-J), bicyclic hydantoin, quirtoline. t :tr4~latcirastlniracsl at .. and analogues thereof, that have if i'1vO androtgenie and anabolic activity of a nori-steroidal ligand for the andre d en r ;t e;pÃor.

loos-si In certain embodiments, the selective estrogen receptor modulators (SERMs) 1. include hart are not limited to_ :non steroidal ligands of the estrogen receptor that are capable of inducing a number of conformational changes in the receptor and thereby eliciting a vartebv of distinct biological profiles (e.g. prevention of estrogen-induced inflammation), and estrogen antagonists (steroidal, non-steroidal) irregardless of receptor atTtiniÃvv. In certain embodiments, the .PRG4 inducing; compounds also include aromatase 20 inhibitors, antiproteases, pro-in[lanamators cs tokinc antagonists, such as an a anti-TNFtx anti 1odv. a soluble T ` 'F receptor. or an IL-I receptor antagonist, cvtokilae release inhibitors, NF-tc-B in hribitors. cvtol ines (c.g. TGFa ). anti-inflrrnrnratorv agÃents, such as cyclo`pori.ne A, omega 3 and 6 f atty acids. or proteasou.ae inhibitors.

(haft] In certain embodiments, the pharmaceutical composition of the present invention 25 comprises a therapeutically effective concentration of a 1?RG4 inducing compound in the range of about 000014).1% vv/v, in combination with a therapeutically effective concent.tation of PRG4 in the range of about. 10-10,000 aSIli __. In certain embodiments, the pharmaceutical composition of the present invention further comprises a t erapeaa:tically effective concentration of one. or more hyaluronic acid or salts ther .of, in 30 the mange of about 10-100,000 tag/ml_ . In certain embodiments, the pharmaceutical composition of the present inventic3n further comprises a therapeutically eft ctiye concentaation of one or more surf.<acc active phospholipids. such as !,-cxr Ã.il l3a~itt~~il~lxessll~,ticl ichcal.itaa (DPPC) phosphaidslchoin: (K), phosplaatidyle..thaanolaamtine (PE), sphingom elfin (Sp), n :utral or polar lipids, in the range S of about 10-1Ã3,00Ã3 tg!mL. The present invention provides that the combinations of a PRG4 inducing compound and PRG4, and other modulators or boundary lubricant molecules allow the direct transport of I'R 4 and other boundary lubricant molecules to the ocular surface cells, where the PRG4 and houndar .- lubricant molecules tend, to raggrogate, and provide ~a. pharmaceutically efficient carrier to the cornea and col ju acti to for efficient modulation of boundary lubrication.

ltHa17j In certain embodiments., the pharnr,accuticai compositions described herein compriso a residence-time increasing agent that increases the residence time of the PR:G-4 inducing, comps: und. on the ocular surface in some embodi.Ã e ts. the residence-time-incr .asinay agent is present in all amount such that when the pharmaceutical composition. is a administered to he surface of an. eve Of an individual, a therapeutically effective amount of a PRC4 inducing compound described herein is retained upon the surf-ace of the eye. In certain embodiments_ the as silence taraa~. rare re asaai agent is selected and/or is present an an amount such that the therapeutically of cove amount of the P.R04 inducing;
compound is retained on the surface of the o e for any thei~apeutically ef3cctiyc period of tisane, at least 1 minute. at least 2 minutes, at least 5 minutes, at least 10 minutes.
at least 15 minutr'-:.s, at least 20 minutes, at least 30 minutes, at least I hour, or more, In c :xtain embodiments- ophthalmic all , acceptable residence-tinge increa.silw, agents or naaucoudhesives may include, by jyax' of non-li.mitin example.
lay dro a par?pc lattc tla ice llarlosE _ carhoxy,an th y lc llulose_ carborn r (acrylic acid polymer), l of (anetlr lrarcil3ae~ .late ), polvacrxlaamide, polycarbophil, polyethylene oxide, acrylic acid,{haatvl acrvlatc copolymer, sodium alginate, dextran, or combinations thereof. The present invention encompasses any high molecular weight polymers that would increase the time that the l>RG4 inducing compound remains on the sus fiac:e of the e e.

lta0181 The. phaarmaaceuticaal composition oFthe pr .sent invention m:a also comprise one or more ophtl alrnically acceptable agents selected Ã=rom the group consisting of an ophthalmically acceptable demulcent, ophthalmical.ly..ac;ccptahlc ;xc.ipiCnt, ophthalmic ally {
acceptable astringent, olxhtlxahnically acceptable vasoconstrictor, arrd ophthaalmicalla acceptable e ollient, l0Ãr191 Exemplary oplathaltn.ically acceptable demulcents contemplated in the present invention include, but are not lira ite to, carboxv metths Ic ellit]osi sodium (e.g.. about 0.2 to 2.5'a,%c, w /v), h droxycthyl cellulose (e.g.. about 0.2 to 2.5" o lr;wpr'ornellose: (e.g., about 0.2. to 2.5% w/v); methyleellulose (e.n.. about 0.2 to 212.5% xvl )~ dextran 70 (.g., about (3.1.: ~U:% %: , gc aiirr (e.g., about tp.4)1~c, rv ~ v ), ~ ~, cc:rirx about 0.2 to 1%) polyethylene glycol 300 (e.g., about 0.2 to 1',% w! ), polyethylene g vecol 400 (e.g., about U to 1% ,v v), pohsorbatc: SO (e.g a.?c-out 0.2 to 1% ks/y)t prop lcne ghh ;ol about 1.0 0.2 to w'v), pzolvvin .l alcohol (e.g., about 0.1 to 4%'%f vv/3) povidone (e.<n.s about 0.1 to 2% w v), Exemplary ophthalmically acceptable .ecilxrcrrts/e rnc,lli rrts contemplated in the pre sent invention include, but are not limited to, anhydrous lanolin about I
to llli4) vs {vl, lan.olin (e.g., about I to 10% iv)., light mineral oil = about 50%
VIVA), mineral oil 4e.g.. = about 50% wvl`v), paraffin (c,&, = about 5% w/v)- petrolatum (e.g.. = about.
100% w' ), white ointment about 100% sv/ ), white.. pctr olaturn (e.g., =
about 100% xv/v), white wax (e.g., = about 5% w/ ,v), yellow -wax (e g., = about 5%
vv/'O. All ese:ragplary ophthalmically acceptable astringent contemplated in the present invention includes, but is not limited to, zinc sulfate (e g., about 0.25% Exemplary ophtbalmically> acceptable vasoconstrictors contemplated in the 'p'resort invention include, but are not limited to, e:.plxed.rine:. hydrochloride (c. , about 0J2')',,/,,, v), naphazolin+
hydrochloride (e., about 0.01 to about 0.03% w/v), phenylephrine hydrochloride (e.g...
about 0.08 to about 0.2% vv/v)., and tetr hvdrozolirre hydrochloride (e.g., about 0,01 to about 0.05% w/v), i00201 In some of these. embodiments, the demulcents, excipients, astringents, vasoconstr-ic.tors. emollients and electrolytes provrefc a means to deliver the PR64 inducing compound and the PRG4 protein in an ophthalmically acceptable manner.
Ophthalmically acceptable compositions are suitable for topical application to the ocular surface if they lack unacceptable eye toxicity, burning, itchiness, viscosity, blurred vision, etc, upon application.

100211 In certain embodiments, the present mention provides a pharmaceutical composition suitable for topical application to an ocular- sin face:
comprising a them-pi uticailly effi ctive Concentration of a P.RG4 inducing compound and PRG'4 protein suspended in a phosphate buffered saline solution or an oph-thalmically>
acceptable balanced salt solution comprisma, tear electrolytes, which iuclri.de, but are not limited to, sodium chloride ( u,. about 44%-54% mole fractions, potassium chloride (e 4?., about 14`x'rs mole fraction), sodium bicarbonate eabout $~ir't3"l - ';~` ~ !~
(e.g., ~T nic?le .l'i'aCi7C)f'E potassium bicarbonate (e.g., about O%o-41',.i, mole fr~action), calcium chloride (e.g..
about 0%4'%/%% mole fraction), III chloride (e.g.. about t} f,Y-4% nao:9l(, fraction), trisodium citrate (e.g., about O% 4% mole friction), hydrochloric acid. (e.g., about 0%-"0'-/" mole fraction) or sodium hxdroxide (e.g., about mole fraction). In one embodiment, the carrier could be formulated to generate an aqueous electrolyte solution in the 150-200 m )'I range.
10Ã 221 in certain embodiments, the present invention prole isles a pharmaceutical composition suitable. for topical aappl ratio ri to an ocular Surface comprising a therapeutically effective concentration of a PRG4 inducing compound and PRG4 suspended in an ophthaalmically acceptable balanced salt solution comprising at least three electrolt`tes, including but not limited to, sodium chloride (NaaC'1) 0.64%, potassium chloride (KCI) 0.075`:%~, calcium chloride dilivdrate (Ca02-2H2O) 0.04%.
magnesium chloride hexahvdrate= (MgCl2-6H.20) 0.03` . sodium acetate trihydr'ate (C2H3NaO')-') l20_t 039 %N~ , sodium citrus dehydrate. (C6f-15Nla3O7N2t-120) 017%, sodium hydroxide and/oi- hydrochloric acid (to adjust pH to approximately 7.5) with an osmolarity of approximately 300 n-iOsms/-L.

100231 In certain cn'ibodimen, the present invention provide=s a.
pharmaceutical composition suitable for topical application to an ocular sur hc.e comprising ai therapeutically of ective concentration of a PRG4 inducing compound and PRG4 suspended in an ophthalmically acceptable balanced salt solution, comprised of sodium (Na-i-) of ai proximately 128 m.M, potassium (K.-I-) of approximately 24 mM, chloride (Cl-) of appioximate1 ' 1 1: in MM, calcium (Ca2+) ofappr'oximateli 0.4 rnl+vi, magnesium MO-) of approximately 03 mM, HCO3- of a appr'oxi.mately 5 nAA citrate of approximately I
nmM, phosphate . of apps-oximately 14 nrM, acetate of approximately 15 mM, and sodium hydrc-oxide aai.dlx_ir h drochlovic acid (to adjust pH to approximately 7.5) ti with an osmol irit of approximately 300 mOsms,,L.

ct [(H)241 The presc ut: ins c aitioa3 further prop iclc s a. method for treating ocular lubrication deficiency, or symptoms associated therewith, in an individual in need. The method ca_ mpri.ses topically administering to the ocular surface of the individual in need any pharmaceutical composition described laere:i , in some embodiments, the pharniaceuticai conaops.ition is one comprising a therapeutically effective concentration of a mduciar compound and a PRG4 protein. In further or alternative embodiments, the.
pharmaceutical comopsitaon is one comprising a therapeutic ally e f'Iec.t { e concentration of a PRG4 Inducing compound and can opiatlb<31aba.icaall accept-able tesidc aace-time incrcaasin a ent. In certain embodiments, the pharmaceutical composition, one comprising the PRG$ inducing compound and the PRG4 protein, is administered in combination with an opl thalrnicaily acceptable formulation comprising one or a or .
oplithalmically acceptable agn.nts selected from the group consisting of an opht aa_imicallt aacceptable demulcent.7 ophthalmically acceptable excipient. ophtliahnica.lls= acceptable a tringent., ophthalmicaaliv acceptable aasoc:caia~trictor, aaaad ophthalm1ca.ils accepLible emollient.

i(f)25] In some embodiments, the pharmaceutical composition, e.g., a composition comprising the P'RG4 inducing compound and an ophthalmicaally a acceptable.
mucoadhe:sive. anent and/or the PR.G4 protein, is administered in combin<aa:tion with can ophthalmicallv:> acceptable solution comprising a therapeutic ally of ective concerti rtion of sodium hvaluronate.. or h valuronic acid, or a surf ace active phospholiliid, as discussed above. In certain ernhc dime.ntsa the pharmaceutical composition comprising the PRG4 i:aarlaacin compound and the PRG4 protein is administered in combination with a phosphate buffered saline solution or an ophtha_alriiicalli' acceptable balanced salt solution comprising one ormaore electrolytes', as discussed above.

100261 In some embodimen s, the present in ention provides a method f' or treating, a deficiency an ocular lubrication or symptoms associated therewith (e.g., dry.
eve)7 due to tear loss or unstable tear film in the ocular boundary- loop, such as androgen deficienc y SjOgaen's syndrome and keratoconjunctivitis siccaa. (KCS). Such method comprises topically administering to the ocular surface of an individual in need the pharmaceutical composition of the present invention.

100271 in certain eembo iments, the Present invention further provides, a method for addressing and treating the conditions associated with ocular lubrication, including unfavorable or deficient ocular lubrication. Exemplary conditions include, but are not limited to, aqueous or evaporative dry eve disease, Sj6gren`s syndrome, kerzatoconjarnctiviti.s sicca androgen deficiency, mei' omian gland disease., estrogen replacement therapy. contact lens wear, refractive surges , allergy. reduced teak film 5 breakup time. gill r F , ocular surface disorxlers, increased protease levels in the tear film and at the ocular surface, chronic infimimation, b perosmolarity, and aging..

l.(Ht281 11e present invention also provides, in certain emhodirn tints, a method of locally inducing PRG4 on an ocular surface comprising topically administering to the ocular surface of an individual in need thereof a therapeutically e;.f lective a mount of any 1.0 pharmaceutical composition of the present invention. e.g.. ar composition comprising a PRG$ inducing compound and PRG4 and/or as ophathaltnically acceptable .rraucoaadlresive a tint. In certain embodiments, provided herein is a method of locally inducing PRG4 on an ocular surface coniprisinca topically administering to the ocular surface of an individual in need thereof a therapeutically effective amount of any Pharmaceutical composition described herein., a therapeutically of ectn e amount of a PR(i4 indu.cin ;
compound being retained on the ocular surface Fora therapeutically of-f~cb e period of time..

BRIEF DESCRIPTION OF THE DRAWINGS

ltH 291 Figure 1 represents kedback loops within ocular surfac : boundary lubric ration.
10030I Figure 2 illustrates PRG4 na.R.NÃA expression in .human corneal epithelial cells.
Human cortical epithelial cells were isolated from the corneoscieral rims of marls. and f%niale.. donors. Amplified samples were screened for the presence of PR G4 products b ,y using an Agiler:rt 2100 Bioanalyzer. Vertical lanes contain: I_.. MW ladder;
1. No template control; 2. Corneal tissue from a 33-tear female; 4. Cultured cortical epithelial cells from a 70-y:ea1r female. 6, Cultured conical epithelial cells from a 53 ear male.

l003f1 Figure 3 illustrates PR(-'a4 mRNA expression n human conj'unctival epithelial cells.
Hr. ma n cortical epithelial cells wet isolated from the corneoscieral rims of male and f male donors. Amplified samples r verc screened for the presence of PRG4 products by using agaarose gel electrophoresis Vertical lassies ce taur. 1. MW ladder, 2.
No template control; 4. Human female cornjunctÃv r.; 5. Human l:m_ e. conjunctiva.

11.
I(H)32.] Figure. 4 illustrates PRO4 rarRNA expression in human conjunctival impression ct t:talog samples. Ccan~tanct:i ral impression cvtolcr_;y samples were isolated from male and female donors. Amplified samples s weae screened. for the presence of PRG4 products by using an Agilent 2100 B oan.alti-zer. Vertical lanes contain: L. MMW ladder; 1-9.
Conjunctival impression. cytology samples; 10. Repeat of human conjunctival epithelial cells (Lane 4 in Figure 3).

l(H)331 Figure. 5 illustrates PRG4 m.RNA. expression in human co.m oscle:ral rim tissue saa~ ples_ L. Human conical epithelial cells were isolated from the corneoseleral urns of male anad finale donors. Anrpli lied samples were screened fs sr the presence of PRG4 1.0 p oducts by using an gilent 2100 Bioanalvzer. Vertical lanes contain: M. 4 ladder; .1.
1-Human liver cDNA standard: 2. Corneoscler al rim tissue from a 24-venr f imde; 3.
Corneoscleral. rim trssue from a. 51 year :fen sale; 4. Hutuan conjunctival epithelial cells.
1(H)34] figure. 6 illustrmates time course of relati e, increase in PRG4 mRNA
in primary corneal ep thel. al cells under the influence of 10 nl i dihydrotesÃostero ne.
Cells were 15 grown ira laeratinocy to serum free media until reaching about S0"/'4 confluence. Cells (n 3 wellsttrc: atza~ezrt:e: l a.aime t) were then incubated with either Ychicle or 10 a M
dih'vdrotestosterone (Di-1T) for cap to ? days. At designated times cells were processed for total RNA isolation and. analysis of PRG4 mRNA mRNA by RU- INCR. The results show that the, D1-1T induces a marled increas ; in P.R04 snRN A levels in prim an, human conical 20 epithelial cells. This androgen effect, relative to the control levels at Day 0, became prominent after 3 (10.3-fold increase). 4 (3.Ã fold increase) and 5 (2.8-fold increase) days ofhormonr.e exposure. 'Ibis D1'1T influence on PRG4 mRNA expression in primary J.-human conical ep.ithelial cells was confirmed in another experiment, Treatment of cells for- 5 days with 1)1-IT caused a 46-fold increase in PRG4 a-raR'' a' content, relative to that of velhicle.
25 treated controls.

DETAILED D 4SCRIPT'ION OFTHE MUNITION

100351 Provided in certain embodiments herein, are, compositions methods fbr-ovating ocular lubrication deficiene (e.g., ocular boundary lubrication deficiency), or Svillptoms associated. therewith, in an individual in need thereof comprising topically administering 30 to the ocul n surface of the ia-adin idu al a pharmaceutical co nposition conrpr icing a thera. peuticaally effective concentration of a PRO4 inducing compound. In specific embodiments, the PRG4 inducing compound is in combination with PRO4. In further or alternative embod.araa%nts, the PRG4 inducing compound is in combination with an opl:athamical.ly acceptable residence-time increasing ag7ent (e.g., an imernt that extends the period in which the PRG4 inducing compound and/'or PRG4 remain therapeutically available in the eye). Provided in specific embodiments herein are pharmaceutical compositions comprising a PRG4 inducing compound and PRG4 in an ophtftalnaical acceptable formulation. In some specific embodiments, provided herein is a pharmaceutical composition suitable for topical application to an ocular surthce comprising a therrapeuticall eft afire concentration of a PRG4 inducing compound e.~ ..
in combination with PRG4) suspended in a phosphate buff ,red solution or at ophthahili.ea lv accept ble balanced salt sohrt.ion., and may also be in combination with one or more oplhthalmicaliy acceptable agents or carrieN selected from the group consisting of an ophthalaxaical.ly acceptable: deratailcent, an ophtla.almically acceptable e.vcipieut. an l phtlaaxlra3icall acceptable astringent, an ophtlaaala3aic al.l acceptable vasoconstrictor, aa3 oplrthalmically acceptable emollie :at. hyalu:ronicc acid. sodium hyaluronate, and surthce :ac:ti e phospholipads.

jwool Provided in some embodiments la rein are pharniacetnical coarmpositiorts_ and me hods of use thereof, for treating a deficiency in ocular lubrication at the ocular surt'ace (e.g.. a deficiency of, such as decreased or undesirable', ocular boundat laubrication). A
pharmaceutical Gi"rr13position of certain embodiment of the present invention comprises as PRG4 inducing compound in combination with an isolated or purified PRG4 protein suspend :d in a phosphaate. buffered solution or an oplithaalmi, ally acceptable balanced salt solution, and further may also be in combination with one or more ophthalillic agents 2: selected from the group consisting of an ophthalmic demulcent, excipient, astringent, vaa.soco.nstructor, and emollient. t.n some embodiments, any pharmaceutical composition provided herein may f irtlher comprise one or more additional therapeutic agents selected from the group consisting of sodium hvaluronate_ surface active phospholipids, and electrolytes in a pharmaceutically acceptable carder for topical administraat.ion.

100371 The present invention provides, in certain embodiments, a pharmaceutical composition for managing decreased ocular boundary lubrication by upregxalKating bounda.r lubricant expression at the ocular surface. In certain embodiments, the pharmaceutical composition upregulates FROG- production by a PRG4 inducing, compound. In certain instances. the upregulation of PRG4 expression Is Specifically localized at the ocular su.ncce (acting on the cotag,unc:tival and corneal epithelium. goblet cells, etc.) and does not .requ.irc; n .odulatio.n of other ocular tissues such as the laac:rimal or me i born ian gland, lttt)3ttl As used herein, a "PRCi4 inducing compoaund" or "PRG4 inducer"
refers to a compound that increases the concentration of PRG4, e.g.. a cà ra pound that is capable of upregulaatinl; PRG4 expressicnr, promoting the biosynthesis of PRG4.
inhibiting the 1.0 degradation of PR[_i4s or the like, including but not limited to, an androgen or aandrogoen analogue, selective androgen receptor r=rmodulaa.tor, selectivc, estrogen r :ceptor modulator, estrogen aarntagonist, aromÃatatse inhibitor. aantiprotÃ.ase, proiu ian-miaÃory. cy'tok n.i antagonist (e.g. selected from the group consisting of antiTT\Fa antibody..
soluble TNFc( receptor. and IL -I receptor antagornist), c toktne release inhi 3:ito:t, antiirrfamEnat<orr l ? ct ttrl:inc (c .g TG _) antiiraflaaaranaattan atgcnt (t. g. cti closporirrc. ., aarx \ t.. eaa:u zl i l l ) kina.se inhibitor, extraccllular--signal regulated kinase (ERK) inhibitor.
mitogen-activated protein (MAP) k.inase inhibitor, and .matrix nietalloproteinase (MMP) inhibitor, omega 3 and 6 fatty acids), NF-K. B inhibitor, or proteasonre inhibitor, and pharmaceuticall acceptable carriers for topical use.

20 100391 In tea ,another embodiment, the androgen or androgen analogue is selected from the group consisting, of a l ,,cr-metlr l-1 rr -lr .dro -? ova-~rÃ
4araclrostiara- -oaac 1Ã riv a.trve.
.a nitrogen-substituted a androgen.. a testosterone derivative, is a 4..5Ãx-dihvdrotestostero.n derivative, a 19-nortestosterone derivative, a 173 1r a tÃ>s~~ o-ara~trostarrc derivative containing a ring A unsaturati.on and a structural subclass of androgens comprising 25 androgenic compounds with unusual st.ructuml features.

100401 In another prefer red embodinment, the selective aandrogera receptor modulators (SARI' s) are selected from a group consisting of aryl-propion.amide (e . S-3-(4-aacct larxaiaxes-lheara~~.t)r?-1r~drtpsy-?-mcthsl v(4ra~itro-34rifiuorometh i phensil-propionauaide [S-4], or ~;-3-( Ilaaofc~plak:rac~ )- :-h~ tlr~ _? aaaetiati i-l -( -.ta: fro-:3_ 30 trifluo:crorm etlat l-phenvl)--prc)piornammmi.de [_S-1.1). bicti clic ht dar:ntoiri, cluintiol:ine, and tetrah clroÃfaziaxoline arrralo ua s that have in-viva androgenic and anabolic activity of a non-steroidal l gaud for the androgen receptor-100411 In vet another preferred embodiment- the selective estros?;ela receptor modulators (SERMMIs) are non.-steroidal ligands of the estrogen receptor that are capable of inducing a number of conformational changes in the receptor and eliciting a variety of distinct biologic profiles. Preferably. the SERMI.s are those that prevent c st:togen-induced Inflammation in ocular surface tissues'. In certain preferred embodiments, the estrogen antagon tsts are steroidal or non-steroidal compounds independent of receptor affinit"W";
100421 Another embodiment of the present invention provides for the methods and pharmaceutical compositions as entioned above for nianagin , decreased ocular boundary lubrication by modulating hypcrosiL olarity at the ocular surface. By interrupting the feedback mechanisms, i >hich prevent secreted components from reducing friction coefficients and mitigating shear stress, the present invention includes pharmaceutical ca_+mpositions for managing decreased boundary lubrication by modulating osrnolarity at the ocular surface.

[((431 In another embodiment, the present invention also provides a therapeutic composition, and method of use thereof to manage and alleviate undesirable conditions for ocular boundary lubrication by compensating for alterations in sex steroid expression at the ocular surface. Androgens inhibit aromatization in synovial cells when their concentration is sufficiently high. Testosterone antagonizes the effects of IL-l. on both proteogivcaan loss and proteogitican synthesis in cartilage. Dehydre epiandrosterone, all androgen precursor, decreases knee joint m ellin during acute and chronic antigen-induced arthritis (AIA), as well as histological signs of inflammation and joint destruction tltn irag chronic ALA (9j. Androgens also appear to protect cartilage from ira.lcanr.mation-715 induced brL. akdon n. This finding supports :a. pathoog nic role for hs>l o anclrogemsm in rheumatoid arthritis and suggests that to ag-term androgen replacement may help pre sent joint damn e and disabalit y Thus, in one embodiment of the invention., the proinflammatorv cytokine-induced effects on sex steroid imbalance and associated inflammation at the ocular surf ace may be countered the by administration of androgens, selective estrogen receptor modulators. estrogen antagonists and arom atase i.niaibitors.

lDH0441 In some embodiments, the present invention also provides a.
pharmaceutical co.n.tposition, and method of use thereof, to manage and alleviate.
undesirable conditions for ocular boundary lubrication, by compensating for the inflammation- and protease-induced reduction iii the hc~attarla:t -Ita.l~ric atita47 abiliÃv of svnao 0al and tear à ud by the administration of factors that sup r s .inf aaaaaa anon and interfere with protease activity.
ltat45) Certlln embodiments of the present invention Provide therapeutic compositions, and methods of use thereof to manage and alleviate undesirable conditions for ocular boundary lubricrttion, such as chronic inffantataation and hypeta smodarity that result from androgen defici.ca cy- estrogen replacement therapy. contact lens wear, compromised tear 1.0 fil:nm, allc.rg\ aging, ocular surface diseases, and increased proton e level's in the tear film and at the ocular sur ace. In one embodiment, mOdulatiort of PRG4 regulation On the ocular surface promotes favor able conditions for proper boundary lubrication by interrupting the central positive feedback loop through reduction of shear stress at the ocular surface.

15 l0O46l It should be noted that the importance and the mechanism of ocular bound.aar lubrication has not heretofore been recognized width n the ophthalmic community. For years, the scientific consensus within the orthopaedic research community was that hydrodynamic lubrication was by liar the dominant mode of lubrication for articular cartilage, and that boundary lubrication was simply an afterthought. Moreover, those researchers studying boundar lubrication at cartilage surfaces. suggest that boundary lubrication is likely only important under `high load and loo t.-docity."
which are opposite to the. conditions at the ocular surface, where there are relatively lows-axial loads and rela:ttvely fast sliding velocities. See, e.g., (10). Moreover, boundary lubrication involving the comcal glyocalys has not heretoffbre been considered Jay t a). compared purified lubricating factor from bovine synovial fluid to "naucinous glycoproteiat from human submandibular saliva and stimulated tears," and concluded "mucin secreted b ~
the lacrimal gland did not lubricate," overlooking the possibility that the corneal epithelium was a source of lubricant or that boundary lubrication was an important contributor at the ocular surface. See, e.g., (11). The most recent mathematical models of tear film dynamics also ignore. the possibility of boundary lubrication, claiming a "lubrication approximation"
for the height of the tear film such that " the mucus layer on 'the cornea can be taken to provide a nonslip surface for the aqueous film" and that it should be noted that the model only predicts the evolution prior to the Itear filml thickness reaching some critically thin val tie at which the model breaks down. See, e.g., (12).

lt)Ã147l There. is a need to mataaage ocular lubrication and protect the cornea and conlutactiv a against significant shear forces generated f rotn the undesirable conditions described herein, including, byoray of non-.limiting example, aqueous or evaporative dry s e disease, Sjo .ten's syndrome.. ke:ratoconiunctivitis siera. androgen deficieancy, meibomiata gland disease. estrogen replacement therapy, contact lens wear, refractive `ta.r ;cry, aallerg\ tv hired tear film break-up tune. atllergs , ocular surface discurdcrs, 1.0 incre<ased protease levels in the tear film and at the ocular surface, chronic inflammations hyperosmol aritv., and aging.

)48i in some instances, the loading of cornea and con unetiva. is likely dominated by shear forces. hi certain .instances eyelid blinking, as r >ell as contact lens wear, generates significant stress upon ocular surface epithelial cells, and this is especially true in the presence of a compromised tear film. As shop n in Figure 1, it is suggested that increased shear stress leads to tear film instability, evaporat:iv tsar loss, ht pcrosmolarity, changes in swelling pressure and a feedback elevation in shear stress. In some instances, increased shear stress is also tic-ought to promote inflammation, androgen deficiency and decreased expression of proteoglycans. In certain instances increased shear stress and its sequelac 710 may. over ti tae, lead to as loss of boundary Iubrication at the ocular surface.

100491 A deficiency. in ocular lubrication and symptoms associated therewith can be determined by any suitable ro thod. In some instances. a deficiency in ocular lubrication and symptoms associated. therewith is defined. either qu alitativel (c, g., aY
feeling of lore lubrication, dry- c,r'c, discomfort, etc_ or qu antitativei (e.g., measured through mechanical, biochemical, electrical, optical or other methods of quantitatir c assails).

l00,50l 1n certain instances, in undesirable conditions for ocular boundary lubrication, such those resulting from aqueous or evaporative dry eye disease, S_jOgren`s syndrome, Li tocoi Rjunctn iti sicci, androgen deficiency- inciboriat3 gland disease, estrove,:n replacement therapy, contact lens wear, refractive surgery. allergy, reduced tear film 3F: breakup time. allergy, ocular surface. disorders, increased protease levels in the tear film and at the ocular surface, chronic inflammation, .hvpc osmolarit :'., and a gin ;, a compromised tear film will exist. In some of these situations, increased evaporation may preclude; efficient fluid film lubrication, but allow boundary lubrication and a molecular sacrificial mechanism to reduce shear stress at the cell surface. Certain embodiments of the present invention provide that therapeutic regulation, replenishment and enrichment of boundary lubricant molecules at the ocular surface would interrupt the feedback loop through which d w unfavorable conditions as soc.tati d r.vltla a deficiency ia ocular lubrication promote ocular surface distress.

100:511 In certain instances, and. as provided, herein, PRG4 protein plays a critical role in 1.0 the eye as .a boundary laub.ricaant. In some instances, this secreted give oprotein protects the ocular surface to Protect the cornea and conjunctiva against significant shear 'forces generated during an eyelid blink, contact lens w year, and any other undesirable ocular boundary lubrication caused b chronic inflammation and atvperosmolarity that result from dr-m eve disease. and:ro ge.n deficiency, estrogen replacement therapy, compromised tear 15 film, allergy, aging, ocular Surface diseases, and increased protease levels in the tear .film and aat the ocular surface.

1100.21 In another- e: emplay embodiment, the present invention features a sacrificial mechanism for ocular boundary" lubrication, thereby surface bound receptors rzv rsiill bind one or more gel forming or surf actint constructs. Ill some llistzmce.s.
the 20 or surfactant constructs detach during at shear event, thereby preventing the shear stress from reaching (or reducing the shear stress reaching) the epithelial surt`rtce. In certain embodiments, following the transient shearing event, the gel for -n. rig and surlactaatit constructs, allowed to return to their undisturbed equilibrium, rebind to the surface bound aecepto s, in some en bodiraments, the entirc construct can detach during;
shear. In certain 25 instannces, the thermodynamics of this equilib"rium can increase the probability of release from the receptor ~\.'ih increasing shear aaamplitude, but any one, association may be easily!
reversible.

lattrs3l Anti- pharmaceutical composition of the present invention a composition comprising a PR64 inducing compound and PRG4 protein suspended in a phosphate 30 buffered solution or an ?phthalmicaally acceptable balanced solution) Is applied topicadly to thÃs ocular surface., wherein the..PRCi4 indLICIng c.ompou:nd uprLgulafes PR:.G4 protein expression and localization on the ocular surace, where the. PRCi-4 associates, binds to. and acts as a surface bound receptor that is allowed to interact with endogenous proteins and proteoglhcxis within the tear filan to establish a sacrificial mechanism to reduce the friction during eyelid blinks at the ocular surface. prevent protein adsorption at the ocular surface, and reduce dry spots caused by tear film instahrliv .

I04r541 In another embodiment of the current in ention, arty pharmaceutical composition described herein (e.g-, a composition comprising a. PRG4 inducing compound and a PR.G4 protein) may also be in combination with one or more of ttalurouic acid and phospholipid constructs. In certain instances of this embodiment, PR0 l acts as the surface bound 1.0 receptor that interacts x 6th the exoz enously supplied .hyaluronic aacrd and/or phospholipids to establish the sac rificial mechanism to reduce the friction during eyelid blinks at the ocular` surf acct;, prevent Protein adsorption at the ocular su fiace, and reduce dry spots caused by tear film instability. In this c.mbodime t., the hyalu:ronic acid and phospholipid constructs disassociate from. the PRCi4 during a shear event. In y'et another embodiment..
15 the dntire construct detaches during a shear event to prevent the shear stress from reaching the epithet tuna .

lttÃr. si In yet another embodiment.. functional fragm nt:s. multime.rs (e.g,.
dimers, tuners, tetramers, etc), homologs or orthologs of PRG4 act as the surface receptor andI<r gel forming, constructs in the sacrificial mechanism. Functional fragments and homologs of 20 PRG4 include those with a fewer repeats within the central naucinlike KEP
PTT-.repeat domain, glticosylated and non-glNlcosylated forms of the protein, splice variants, vecombinant forms, and the like.. lubricating fragment of PRG4 exhibits at least 20%.
30%, 40%,, 50%,, 60%,, 70 ,'s,, 80%. 90%. or 5' , of the ophthalmic lubricating effect of human P1 4, as rah, arsured qualitatively, cart cliattarcallk, opticalll, elcctricall or by 25 biochemical assay, 10O56I As used hereitr, the terra "P'R,54." ::PRG4 protein.` proteogltican 4," and lubricant," are used interchangeably. PR..G4 is used herein also to encompass the term megakamocyte stimulating} factor (.SF'), that has been accepted for the UCL/i-ICGNC/1-IUGO Human Gene Nomeaclatur d r.ta base, and superficial zone protein 30 (SZP). The PRG4 or lubricin protein (used interchangeably herein with lubricin proteoglvcan) as used heroin refers to any isolated or purified native or recombinant f9 htbricin proteins, homologs., fractional fragments or motifs, isofonxts, and,'or mutants thereof. In certain eanbodime.nts, the isolated or purified PRG4 protein comp ises an amino acid sequence for a human natisve or recombinant 1tibricin protein, In other embodiments, the isolated or purified PRG4 protein comprises an amino acid sequence encoded by 1-wk-4 gene cx.ons that encode the full length PRG4 protein or isoforms' primary structures. The proteoglycan 4 (pr-;4) gene contains 12 exons. The PRG4 protein used herein comprises an amino acid sequence eancoded by Tr 4 ygene esons I-12. more. preferably, e.xons 6-12, and most preferably, exons 9--1.2.

l0O 7l As used her;in, the PRG4 protein incluc.ies any PRG4 proteins narti ktaoi gin, or later 1.0 described. In certain c Ilbodiinents, a preferred PRG4 protein an ino acid se hence is provided in SEQ ID NO. 1. The PRG4 protein shares the. prtn aty an no acid structure of any known PIt(14 protons or isof"ot' ns with at least 60'-?% ho nology. pr :ferably 75'%4) llotl ology, more prefe.:rablv 85`fi~ 90%. 95%'r3, 96%. 97%''r}, 9or more homology. III
certain embodiments, a preferred PRG4 protein has an average molar mass of batZ ,een 50 15 kDa and 400 kDa, comprising one or more biological active portions of the P'R(4 protein, or functional (raa awnts, such as a tubricatin trFagtl ent, car a la atatolo die roof I(ximl As used herein., the PRG4 protein comprises a biologr.ical active portion of the protein. As used herein, a "biologically active portion" of the PRG4 protein istcludcs a f factional fragment of a protein comprising amino acid sequences sufficiently 20 homologous to, or derived from, the amino acid sequence, of the protein, which includes f wer amino acids than the full length protein. and exhibits at least one activity of the full-length protein. Typically a biologically active portion comprises a functional domain or Motif with. at least one activity of the. pro-win. A biologically active portion of a protein can be a polypeptidc which is, for caample, 10. 23, 50, 100, 200, or more. amino acids in 25 len<gth. In one embodiment, a biolog tally active? portion of the P G4 protein can be used ,as a therapeutic agent :clone or in combination with other therapeutic agents for treating undesirable or decreased ocular boundary Iu:brication.

joOS9l The nucleic acid and amino acid sequences of several native and recombinant PRG4 or h. bricia proteins, and characterization of the PRG4 proteins and various isofbiins 30 are disclosed in. for instance. U.S. Patent Nos. 326.558;
6.4x3.142:7.030.223:7.361.738 to Turner et at, and U.S Patent Nos. 6.743.774 and 6,960,562 to Jay et al-U.S.

Publication No. 20070191268 to Flame ,:v et al. also di ck-,se recombinant PRO4 or lub.ricin molecules useful in the present invention, lotna o Methods :tor isolation, purification, and recombinant expression of a.
PRG4 Protein are a.vcll known in the art. In certain embodiments. the method starts with clornin and 5 isolating niRN l and eDNA encoding PRG4 proteins or isoforms using standard molecular bio.logti techniques. such as .PC.R or R"T-PCR. The isolated cDNA encoding the protein or isofornm i then cloned into an expression vector, and further transfornned and expressed in a host coil for producing recombinant PRG4 protein.

0Ãrt~tl .~ .s tts d herein. '`rc:i:.ombr ant`, re.t rs ttr a p~al~trcrcl oti e sxntla;sized ter others isi 10 manipulated in vitro (e. gA.. "recombinant polvnuclcotidc" ), to methods of using recombinant poll rrue(t.otid.es to produce gene products in cells or other biological systems, or to a peal;=pept:idL ("recombinant protein") encoded b a rccomhirtaant polsnucleoude.
"Recombinant" also encompasses the libation of nucleic acids having various coding r gior:ns or domains or promoter sequences from dif .rent sources into an expression 15 cassette or vector for expression of., e.g., inducible or constitutive expression of a f=usion protein comprising an active domain of the. PRG4 gene and a nucleic acid sequence amplified using a primer of the invention.

i(rtr621 In certain embodiments, the. PRG4 protein encoding nucleic acid may contain one or nor. mutations. deletions. or trtscrtions. in such embodi.naents_ the P RG4 Protein 20 encoding nucleic acid is at least 60% homologue preferably 75% homology, more preferably ii5SN,, 90%, 95%. 9C~ ,, 97 d%, c) tsar, 995,'. or more honmology, to a wdd ty e PRG4 proteia encoding nucleic acid.

l0#t631 As used herein, the tern cl).N A,`' includes .DNA that is complementary to mRNA
molecules present in a cell or organism mR lr\ that can, be converted into eDNA with an enzyme such. as reverse. tr rnscriptarse. In certain embod_ime ats, the eD.NA
encoding PRG4 prote:in IS isolated from PRG4 mR.N.A. expressed. in humaan corneal or conjunctival epithelial cells using an RT-PCR method well knor.wwn in the. att.

[0641 As used herein, the terms ' pola.nrarclt rrticl~ " -`nucleic acid'uucl .otide, " and oligonuclcotide" are used interchangeably, and include polymeric torus of nucleotides of any length, either deoxvribonucleotides or ribonueleotides, or analogs thereof.

2I.
Polvn cleotides may have any three-dimensional striucrure, and may perform an"., f tnction..
l nowvn or unknown, The fbliovwig, are non-limiting examples of polynucleotides: a gene or gene Ã'ragment. exons, introns, messenger RNA (mRNA). transfer RNA, ribosomal RNA, ribof\ me , DNA-, cD_NA, igonomic D-N:\, recombinant polynuclcotides, hrarnched polynucleotnles, plasmids, vectors, isolated DNA of any sequence. isolated RNA
of any sequence, nucleic acid probes, and primers. Polvnucleo-tides may be naturally-occurrill",.
synthetic, recombitaa.aat of arA cotribinat ort thereof.

1001 A polynurcleotide max' comprise modified nucleotides_ such as methylated nucleotides and nucleotide analogs. If present. modifications to the nuclecitide structure 1.0 may be imparted before or after aassembl of the polymer. The sequence of nucleotides naay be interrupted by non-nucleotide components. A polvinicleostide may be further modified after polymerization, such as by conjugation with a labeling component. The term also includes both double-and single-stranded molecules. Unless other wise specified or required, any embodiment of this invention that is a polymicleotide encompasses both 15 the double-stranded form and each of two complementary single--stranded forms kno-'v n or predicted to make up the double-stranded foam.

100661 As used herein, the term ' polymicleotide sequence" is the alphabetical representation of a polynueleotide molecule. A polynucleotide is composed of a specife sequence of four nucleotide bases: adenine (A); cytosine (C): guanine (0);
thymine ('T);
20 and uraacal (U) in place of divna.ine wvhen the polvnucleotide is RNA, instead of 0 A. Tla.is alphabetical representation can be inputted into databases in a computer and used for bioinli nnaatics applications such as, for example, Ã inctional genomics and homology searching"

1(067] As used herein., the term "isolated polynucleotideleDNA" includes polynucleot.ide 25 molecules v hich are, separated from other polvnucleotide molecules which are present in the natural source of the. polyuueleoti.le. For example, wvah regard to genomic DNA.. the term '-Isolated" includes polytaucleotide molecules which are separated from.
the chromosome with which the genomic DNA is naturally associated. Preferably, an `isolated" polvnucleotide is free of sequences which naturally flank the polvuucleotide 30 (i.e., sequences located at the 5, and ?' ends of the poll nucleotide of )nterest) in the {ge:nomie DNA of the organism from which the po nucleotide: is derived. For e\anaple, i:n various embodiments, the isolated polyaauclcotide molecule encodin the PRGF4 Protein used in the. invention can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb.. 0.5 kb or 0. l kb of nucleotide sequences which naturally flank the polynucicotide molecule in genno nmic DNA of the cell from which the polvuucleoticle is derived, .Moreover. an `isolated"
polynucleotiale molecule, such as a eDNA nm.olecule. can he 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.
10O681 As used herein, a :.gene" trrcludes a poltinucleotade containing at least one open Leading f flame that is ciipable of encoding a particular poltpeptsde or protein after being 1.0 transcribed and translated. Any of the polvnucleotide sequences described herein mat also be used to identify larger fragments or full-length coding; sequences of the gene with which they are associated. :Methods of isolating larger .fragment sequences are known to those of skill. in the art. As used herein. a "native or naturall -occuraing"
polynucleot:icle molecule includes, for example, an RNA or DNA .molecule having a nucleotide sequence 15 that occurs in nature (e.,r~,. encodes a natural protei.n).

10Ã69l As used herein, the term '`polvpcptidc" or "protcirr" is interchangeable, and include r a compound of two or more, subunit amino acids, amino acid analogs, or peptidomit-notics. The subunits may be linked by peptide bonds. In another embodiment, the subunit may h linked b-,., other bonds, e.g,, ester, ether, etc. As used herein, the teen 20 `'amino acid" includes either natural rand,.{car unnatural or synthetic amino acids, including glvcine and both the D or L optical isomers, and amino acid analogs and peptidonrimetics.
A. peptide of three or more amino acids is commonly referred to as an oligo ept de.
Peptide: chains of greater than three or more amino acids are referred to as a poly>peptide or a protein.

2? ltrrr~t-l In certain embodiments, the PRG4 protein used herein refers to PRG4 proteins or various homologs or isoforms thereof. that are naturailti or reco binantly expressed in humans or other host cells. As used herein, "express" or "expression" includes the process by which polynuciecotides are transcribed into RNA and/or translated into polype ptides. If the poll=nucleotidc is derived from genoinic DNA, expression may include splicing of the 30 RNA, if an appropriate eukar yotic host is selected. Regulatory elements required for expression include promoter sequences to hind RNA polvmerase and transcription initiation sequences for ribosome binding, For xampli:., a bacterial expression vector includes a promoter such as the. lac promoter and for transcription initiation the Shirae-I)aaigirrio sequence and the start codon AUG. Similarly, a eukaryotic expression vector includes a heterolo ous or homologous promoter for RNA pall:meaa e 11, a downstream file ad n '1< tion signal, the start cod on 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. tar example, the rarethods described below for constructing vectors in general. As used herein, the term "vector"' includes a self rcp.licating nucleic acid m l.ecule that transfers an inserted polmnucleotide into and/or between host cells. The term is intended to include vectors that function primarily for insertion of a nucleic acid molecule into a cell, replication vectors that function primarily for the replication of nucleic acid and expression vectors that. function for transcription and/or translation of the D.N.A or RNA.A.lso intended are vectors that provide more than one of the above function.

l 71j As used heroin, a "host cell' is intended to include any individual cell or cell culture o, which can be. or has beers, a recipient for vectors or for the incorporation of exogenous po.ivnuclecotides and/or po.ivpeptides. It is aalso intended to include progeny of a single: cell. The p ogwny may not necessarily be conapleteiy identical (ira morphology or in <genomic. or total DNA complement) to the original parent cell due to natural.
accidental, or deliberate mutation. The cells maa he proltiarvotic or eukarsolio, and include but are not limited to bacterial cells, yeast cells, insect cells, animal cells, and mammalian cells..
including but not limited to murane, .rat, simian or human cells. As used herein, a "host cell.' also includes genetically modified cells. The term genetically modified cells"
includes cells containing and/or expressing a for,.ign or exogenous gene or poly-nucleotide 2: sequence which in turn modifies the gc rrot lae or Phenotype of the. cell or its progeny.
"Gene..ticallti modified" also includes a. cell containing or expressing a genc or polnucleotide sequence which has been introduced into the cell. For example.
in this embodiment, a genetically modified cell has had introduced a gene which gone Is .11.'-,o endogenous to the cell. The terns t;cnetically modified" also includes any addition, deletion, or disruption to a. cell's endogenous nucleotides. As used herein, a ``host cell" can b : any cells that express a human PRG4 protein.

lD072j As used herein, `'homoloes" are defined herein as two nucleic acids or peptides that have similaar, or substantially ident:ic.al. nucleic acids or amino acid sequences.
vespecÃi-vely. '1hhe 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 s;gUCIICCs. In one of the preferred embodiments, homologs include allelic variants, orthologs, para.logs, agonists, and antagonists of nucleic acids cn odw<g the PRG4 protein [t .g.. SEQ II NO: I).

107,31 As used herein, the term "orthologs" refers to two nucleic acids from different species, but that have evolved from a common ancestral gene b-: speciatic-n.
Nor mall, 1.0 orthologs encode peptides having the same or similar fiunctio:ns. In particular, orthologs of the invention ?. gill generally exhibit at least 30-M7 more prefer. hlt 85-90%
or 90-95%, and most pr ferabhl\ 9'!"%. 96%, 97% 98%, or even 99's,'~ id r.tit\ or 100%
sequence identity, with all or part of the amino acid sequence of a any known .PRG4 proteins SEQ ID NC.1), isofornas, or analogs, thereof, and will exhibit a function similar to these 15 peptides. As also used herein, the term "-I?analogs" re.ui..rs to two nucleic acids that are related by duplication within a genome. Parakigs usual! have different functions, but these functions may be related.

I(H)741 To detenniao the percent sequence Identity of m -o amino acid sequences, the sequences are aligned for c?l?timal comparison purposes (e.g.. gaps can be introduced in the 20 sequence of one polx'peptide for optimal alignment with the other polypeptide or nucleic acid). The amino acid residues at corresponding amino acid positions are then cornpaared..
When a position in one sequence is occupied. by the same amino acid residue as the corresponding position in the other sequence. then the molecules are identical at that position, The. same type. of Comparison can be made between two a ucleic acid sequences.
25 The percent sequence identity between the two sequences is a function of the number of identical positions shared by the sequences percent quence identity = numbers of identical positions/total numbers of positions x 100;. Pae#erably, the isolated amino acid honologs included in the present invention are at least about 50-60%, preferably at least about 60-70%, and more preferably at least about 70-75%, 80-85%, 85-90%, or 30 90-95%. and most preferabl4' at least about W%'), %'), 97 x %%, 98{!%. 99 o, or more identical to an entire amino acid sequence of any known PRG4 protein (e.g..; SEQ ID NO: 1), lD07+1 In certain embodiments, an isolated nucleic acid hortaolog encoding the protein comprises a nucleotide sequence w Which is at least about 40-60{'=%, pref .rably- at least about 60-70%t more pr tRrably at least about 70-75%, 75-80%1, 8M5%, or 90-and even more preferably at least about 95%, 96% 97%, 98%, 99%. or more 5 identical to a nuclcotide sequence encoding amino acid sequences of such PROM protein {e.g., SEQ ID NO: -1 .

ltttt761 the determination of the percent sequence identity between two nucleic acid o peptide sequences is well know ~n in the art. For instatnc e, the Vector N TI
6.0 (PC) software package f la.tfe+a: la _ Bethesda, MD) to determim the percent sequence.
identity beta een 1.0 two nucleic acid or peptide sequences can he used. In this method, a gap opening penalt,\', of 15 and a gap extension penalty of 6.66 are used for determining the percera identity of two nucleic acids, A gap opening penalty of 10 and a gape .tension penalty of l-.1 are used for determining the percent identity of two poll peptides. All other parameters are set at the default settings. For purposes, of a multiple alignment (Clusta:l W
algorithm), the gap opening pe.nalty is 10, and the gap extension penalty is 0.05 with blosurnQ
matrix. It is to be understood that fbr the put-poses of deter ining sequence identity when comparing a DNA sequence to an RNA sequence. a th nmidi ae .nucleot. de is equivalent to a uracil nucleotide.

[W771 Furthermore, the PRG4 protein used herein includes PRG4 prow in encoded by a polvnucleotide that hybridizes to the polynatcleoude encoding PR64 protein-under stringent conditions. .As used. herein, "hybridization" includes a reaction in which one or more poll nucleotides react to firm a complex that is stabilized via hydrogen bonding between the bases of the nucleotiede residues. The hydrogen bonding .ma.?
occur by Watson-Crick- base pairing, l-loogstein binding, or in any other sequenc.:.-specific mtannea.
The complex may co atpnse t vo strands forming a duplcx structure, three or more strands forming a multi-stranded complex, a single self-hybridizing stratad., or array combination of these.. hybridization reaction mays constitute a step in a more. extensive process, such as thw initiation of a PCR reaction, or the enzymatic clean age of a polynueleotide by a ribozynte.

100781 Hybridization reactions can be perfbr e.d under diftat .rat stringent conditions: The present mention Includes pol,-nucleotide capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most prcferabhy highly stringent conditions, to pol :rnucleondes encoding PRG4 protein described herein. As used herein, the term stringent Conditions, refers. to hybridization overnight at 60 C in W x Denhart's soluti in, 6xSSC , 05% SDS, and 100 mw/ml denatured salmon spen.-aa DNA.
Blots are washed sequentially at 62`C for 30 minutes each time in 3xSSC./O.1.`
SDS, followed by I xSSC'/fl..l I% SDS. and finally 0. I;xSS(rt). P,% SDS, As also used her ;irn, in certain cnrbodinreits. the phrase 'strirngent conditions refers to hybridization in a 6xSSC
solution at 6 C. In other embodiments, "highly stringent corlditio:n ` refer to hybridization overnight at 65 C in I0xDenhart's solution. 6xSSC, 0.5% SDS and nags/nit denatured sal iron sperm DNA Blots are. washed sequentially at 65 C
for 30 minute` each time in 3xSSC./{).I";% SDS, fellowved b y IxSSClt;.1% SDS, and finally 0.1xSSC/'0.i% SDS. Methods for nucle c acid hybridizations are well known in the art.
Accordingl the PRC 4 proteins encoded ht nucleic acids used herein include nucleic acid having aat least 60% homology prE fs..raably 75% homology, more prof rally 85%, more pre:terabl.N% 90%, most pr feraably 95%, 961/`r3, 97%7 98%, 99% homology to a polvnucleoti.de sequence that encodes a human PRG4 protein (e. SEQ ID :NO l) or a specific astrtorm or honrolog tlaereof.

100791 Moreover, the PRCi4 proteins used therein can also be chimeric proteth or fusion prateir . As used h.:rein. a "chimeric protein" or fission protein" comprises a first polvpepti.de operatively linked to a second poly peptide. Chimeric proteins may optionally co:nlprise a third, :fourth Or fifth or other polypeptide operatively .linked to a first or second pol`'peptide. Chimeric proteins may comprise two or more different pol peptides.
Chimeric proteins may comprise multiple copies of the same: polypeptide.
Chimeric proteins may also comprise one or more mutations in one or more of the poiz peptides.
2: Methods for making chimeric proteins are well known in the art. In certain embodiments of the present intention, the chimeric protein is a chimera of PRG4 protein with other PRG4 protein, isofoims.

100801 As used herein, an "isr_olatc d" or ` puaitied" protein, polyna.icleotide or molecule means removed from the environment in which they naturally occur. or substantially free of cellular aaa ater-iaal, such as other contaminating proteins from the coil or tissue source from which the protein polv'nucleot:ide or molecule is derived, or substantially- free from chemical precursors or other chemicals when chemically synthesized. The language substant.ialiy free of cellular material" includes preparations separated from cellular components of the cells from which it is isolated or recombinantly produced or synthesized In certain embodiments, the language substantiall free of cellular i lateral"
mcludt s preparations of a PR:Ci4 protein having less than about 30`) (by dad weight) of other proteins (also referred to herein as a "'conÃacaai:naatin prc Ãa:ira"}
rracir preferably less than about 20%, still more preferably less than about 10%. and most prefer tibl less than about 5"==r, of other proteins. When the protein or polynucleotidt is :recoltal iraatltlz produced. it is also pre:feribl-v substantially free of culture medium, i.e.-culture medium ill represents less than about 20%, more preferably less than about 10 ,.x, and most preferably less than about 5% of the volume of the preparation of the protein of interest.

1ttÃ)"4i1 In certain embodiments, the present invention provides a pl-mai ceutical co:niposition suitable for topical administration to an ocular surface Of an individual in need a pharmaceutically effective concentration of a PRG4 i:ndracing compound, an 15 optional mucoadbhesive agent, and, optionally. PRG4 suspended in are oplrthalmiealla acceptable balanced salt s_4tition, and in combination with one or more ophthalntically acceptable aafen.ts. The ophthalmically acceptable aunts can be selected from the group consisting a aan oplithalmically acceptable demulcent, exciplent. astringent, vasoconstrictor, and emollient. As used herein, the. term effective concentration or 20 amount" or "the:rapeuticilly elective concentration or amount" is intended to mean a nontoxic but sufficient concentration or amount of a PRG4 inducing compound., the ]PRG4, or the other therapeutic agents to provide the. desired therapeutic effects. The concentration, or amount that is effective will vary, from sua.blect t i subjeet, depending on the ape and general condition of the individual. the particular agents, and the like. Thus. it 2: is not always possible to specify an exact effective concentration or amount. Hov ever, an appropriate effective concentration or mount in any individual case may be determined 1v one of ordinary skill in the alt using routine experimentation.
Furthermore, the exact effective concentration or amount of a PRG4 inducin conll csaa:nd the PR G4 Prot in, or the other therapeutic agents incorporated into a composition or dosage forage of the present 30 invention is not critical, so long as the concentration is 4 ithin a range sufficient to pennit ready application, of the solution or formulation so as to deliver an amount of the PRCF4 inducing compound, the PRC =4 protein, or the other active aggent that is wvithin a ther~alaealt.ic,allt of ectivc range.

l0Ã82I in certain embodiments- the pharmaceutically effective concentration of a PRG4 mducinn compound is in a range of 0.0001-0.1% wfv, a ad the ph aunaceuticallti effective c:oraccratraatiora of PR.G4 protein is in range of 10-1 0,000 irghnL.
preferably 50-500 lad:.{m.l.:, and more preferably 100-300 l_rg-/mL. As used here, the oplxthalnilcall acceptable agents comprising the ophthalmical y acceptable demulcents, e :cipients, astringents- vasoconstrictors, and emollients that are fully defined in the Code of Federal Renulations 21CFR349.

1(10831 In certain embodiments, the pharmaceutical compositions described herein comprise a residence-time increasing agent that. increases the residence time of the P14(34 inducing compound on the ocular surface. In sonic embodiments, the residence-time-incrc as.ing agent is present in an amount such that when the pharmaceutical composition is administered to the surface of an eve of an individual., a therapeutically eÃi .ctive amount of a PRG4 inducing compound described herein is retained upon the surface of the eye. 111 certain embodiments. the residence-time increasing agent is selected xa:rid,{or is present in an :amount such that the therapeutically effective amount of the PRG4 inducing compound is retained on the surface of the eye for any therapeutically effective period of time. at least 1 minute, it least 2 minutes, at least 5 minutes, at least 10 minutes..
at least 15 minutes. at least 20 minutes, at least 30 minutes, at lust I hour. or more, In certain embodiments. optithaalmically acceptable residence-time increasing agents or mucoadhesivos, may include, by way of iron-larniting example.
hsdroxyprops lmcths lccflulosa;, ca rhoxr mct:hs>lccllulose, caarhor ter (aaer :he acid pola%me' ) paly(metla` lagtc tltaaa r late), poll aacrv-lamide. polveaar .ophil, Viols etlas lean. oxide, acrylic acid./butyl aacr rl.aate copolciner, sodium alginate, dcyturan, or combinations thereof. The present invention encompasses any .high molecular v eight polymers that would increase the time that the PRG4 inducing compound remains on the surface of the eve.

10Ã0841 As used herein, the tam- ''topical administration." is, used in its conventional sense to mean delivery of the co -position comprising the PRG4 protein w id o e or more ophthal.micaally acceptable agents to the eve. In general, topical a administration is achieved throucnlh a liquid formulation for eye drops or lavaa?-e and provides a local efect.

[W851 In certain embodiments, any pharmaceutical composition described herein compose or the aforementioned ophthalmically acceptable agents are or can be combined with one or more of carboy naetla >lt .ellialose sodium (e.g., about 0,2 to about 2.5".%, t .' .
hn droxveth~'l cellulose (t g., about 0,2 to about 2.5% h prt~mell~os : (e.g.
about 0.2 to about 2.5%, met ylcellulose ( g.. about 0.2 to about 2.5'% w/v), dextrin 70 (e.g., about 0.111% ~N _). gelatin (e.g , about 0.01'% w f`v), glycerin (e.g., about 02 to about 1%
w/v) poly ethylene glixcol 300 (e.g. about 0.2 to about l" o wv/ pob ctla .
leetzc ;lycol 400 t2+ F5' j (E . s .. about [7.'1 to about 1% poly sorbate 8 l (e ,:4>. , about 0.2 2 to about I . ~~ .: ), propylene glycol (e g, about 0.2 to about 1% w/v), polyvinyl alcohol (e.g., about 0.1 to about 4 % w v). povidone. (e g.- about 0.1 to about 2% zinc sulfate (L. ., about 0225%
~:% ), anhydrous lanolin (c. ., >about. I to tabout. 10"rc ), lanolin (. :, about I to about 10% Z1'/ )s light mineral oil (e.g, , about 50% w.A), mineral oil (e.g, =
about 50`) w/0-paraffin (e.g., = about whv), petrolatum (e.g., - about I00`3%, vl ), white ointment (e.g.
about 100" w/v)> white. petrolatum f.. h., about 100"o s v/ _), whit . w.a.x (e.g... about :s% iw ev). yellow wax (e.g.. = about 5% w/z-), ephedrine hydrochloride (e_ q.
, about 0.123 ws /v'). naphazoline. hydrochloride (e_ g., about 0.01 to about 0.03% w/ =), phenylephrine hydrochloride (e.g , about 008 to about 0,2% and tetrallvdrozoline hydrochloride (e.g., about 0.01 to about 0.(#5% In certain instances, percent amounts utilized herein are percent amounts by weight.

101)86) in further eanhotlimcntsa any pharmaceutical composition of the present anve:ntio:n (e . a composition comprising a .PR. 4 inducing compound and PRG4) may further co.mpnse a therapeutically e.llcctive concentration of by aluronic acid or sodium la'=aluronate. in flat: Tango of-'10-100,000 t,ig/mL prekiabli. 500-51000 pg mL, Furalics-mcire, the plsais:aa:accratical composition of the present invention may farther comprise one or 2: more surface active phospholipids in the range of .10-10.,000 pg/mL, such surface active phospholipids include.,, but are not limited to, :L' -dipal.mitoylphosphatidv lcholine (DPPÃ;), phospb atidd lchi_iline (PC), pliosphatidti le.tlhanolaxnitie. (11E), and sphingoni elfin (Sp), or other :neutral and polar .lipids. In this embodiment., the combination of the more hydrophobic modulators with the amphiphilic boundary lubricant molecules allows the direct transport of therapeutically effective molecules to the ocular surfaac cells, where the boundary lubricant molecules tend to aggregate. and provides a pharmacemfically efficient carrier for therapeutic compounds to the corneal and conjunctival epithc.liuni for efficient boundary lul ri a[ic>r . For earr pl solubi irx the h dr<?1 hobicar dre ens in OPPC, then co.n.iplexing DPPC., HA and PRG4 in a pharmaceutically acceptable carrier., transports and concentrates t ho androgens at the ocular surf ace cells, where they may most efficiently upregulate boundary lubricant expression.

5 100 ,1471 Tlae f~hararaacc:uti.caal corral?osition o.l` fire present r.rar enticrrt rraaa further coraaprise ona or more pharmaceutically acceptable carriers or vehicles comprising any acceptable materials. and/or any one, or more additives known In the art, As used hereim the term "carriers" or 'vehicle- w for to carrier materials suitable for topical drug, ada ai.a astration:
Carriers and vehicles u.s:titl hereitr. include arnv such materials known in the art, which u e 1.0 nontoxic and do not interact i 0ih other components of the composition in a deleterious maau.ne:r. Various additives, know. u to those skilled in the art= max be included in the c:orrrposition For etanap e, solvents, iracludirrg relatively small amounts.
of Acohol, may be used to solubilize certain drug substances. Other optional additives include opacifiers, antioxidants, fragrance, colo:raant, gelli.ng agents, thickening agents, sta.bihzers, surfactants..
15 and the lilac. Other as erits ma also be added, such as antimicrobial agents, to prevent spoilage upon storage- ix_ to inhi it growth of microbes such as yeasts and smmold .
Suitable antimicrobial agents are typically selected from the group consisting of the methyl and propyl esters of p-hydrotiybe..r zoic acid (i.e., methyl and propel paraben), sodium benzoate. `orbit acid, inaidureaa, and combinations then of. Per eatiorr enhancers 20 and/or i:rr taatiora-rraitiga_ting additives may also be included in the pharmaceutical composition of the present invention.

100881 In certain embodiments, the pharmaceutical co mpositit n of the present invention is prepared in a pharmaceutically acceptable carrier. such as a phosphate buffered saline or an osmotically balanced wilt solution of tear electrolytes, including one or more. of sodium 25 chloride in about 44%.'% to about 54%0' mole fraction, potassium chloride in about 8% to about 14% mole fraction, sodium bicarbonate in about 8% to about 18% mole fraction, potassium bicarbonate in about 0%,, to about 4'%, mole fraction, calcium chloride in about 0% to about 4% mole fraction, naaignesiunr chloride in about 0% to about 4%
mole fraction- trisodiuna citrate in about 0% to about 4% mole fraction, and hydrochloric acid in 30 about 0`4 to about 20% mole fraction or sodium hydroxide in about to about mole fraction. in certain embodiments, the pharmaceutical carrier can he formulated to 3l.
generate an aclueous electrolvte solution in about 150-200 mM ra ge. Other suitable formulations, such as ointments., creatars~ gels, pastes, and the like, suitable for topical aclarmiamistratioan. are also Contemplated in dle present invention. In Certain e bodimeants, elect olytes provide proper osmotic balance when combined with the PRG4 inducing compound anad Optional PRG4 to make a solution. ophthalmicaily acceptable.

1(1)89) ilr.e present invention further provides a method for treating decreased or undesired ocular boundary lubrication, symptoms associated therewith, or a condition that is associated a. ith or causes a deficiency in ocular lubrication, in an individual in need tl ereof, comprising tropically, administering to the ocular surface of the individual in need 1.0 a any pharmaceutical composition described herein. in a spec: fic embodiment, tile Composition Comprises a therapeutically effective mount of a PRG4 inducing compound in combination 'with. PRQ=1. In one embodiment. the method of the present invention comprises topically administering a pharmaceutical composition comprising the therapeutically effective amount of a PRG4 inducing compound in combination with 15 PRG4 that is suspended in a phosphate buffered saline solution or an ophthalmicall a acceptable l alanc cl salt solution comprising one car more tear electrol .tes. In yet other embodiment, the method of the present invention comprising topically administerir:rrg a pharmaceutical composition comprising a PRG4 inducing compound and PRG4 that is formulated in an ophtlaalmic.ally acceptable fFbmi alat:ion comprising one or more additional 20 oplhthalmicall . acceptable agent as discussed <a.bove.

100901 As used herein, the term "treating or treatmeant" refers to reduction in severit and/or frequency of symptoms, elimination of symptoms and/or underlying cause, prey ention o: thy: occurrence of"s mptoms and/ or their a.n rlying :pause, argil .irt p o c:ta ent or r mediation of damage. The tang "trcating or treatment" also encompasses both 25 prevention of a disorder in a predisposed individual and treatment of the.
disordea in a clinically symptomatic individual-10911 In Certain embodiments, the decreased ocular boundary lubrication is caused by increased evaporative tear loss or unstable tear film in the ocular boundary loop. Such decreased or undesired ocular boundary lubrication is associated with aqueous or 30 evaporative dry eye, disease., Sttren s syndrome, keratoconz(unctivitis sicca (KCS), and:roccra dof c:ieraea , rriuiboaarian gland disease, cstra era ielrlaccanerlt tlaerrapt, con tact lei's wear. refractive sur e r , allcnr reduced tear film breakup tirra , compromised tear film.
ocular surface disorders, increased protease levels in the tear film and at the ocular surface, chronic inflammation, hyperosmolarity, and aag-ing. As discussed above, the increased shear stress leads to tear film instability, evaporative tear loss, layperosinolarity, changes in swelling pressure and a feedback elevation in shear stress.
Increased shear stress also prorarote inflaaaaraa ation androgen deficiency and decreased expression of pro eoglycans. Over ttr:aac_ increased shear stress and its sequelac leads to a loss of boundary lubrication at the ocular surfhce. Accordingly, the present invention provides, a method for reducing shear stress by replenishi:n and enriching the expression of proteoglscans, such as PRG4 protein. at the ocular surface, using a PRG4 inducing compound as discussed above, so as to pre vent or increase ocular boundarcy lubrication. A
method of localizing, PR. G4 induction at the ocular surface is also provided in the present Invention.

ltrtt921 throughout this application, viriou publications are, referenced. ' 'he disclosures of all of these; publications and those relhrences cited within those publications in their entireties are hereby incorporated by reference. into this application in order to more fully describe the state of the art to which this invention pertains.

[(H) 931 It should also be understood that the foregoing relates to preferred embodiments of the present invention and that numerous changes may be made. therein without departing from the scope of the .invention. The. imention is .ftarther illustrated by.
the following examples, which are not to be construed in any sway as imposing limitations upon the scope thereof On the contruy, it is to be clearly understood that resort rnaay. be had to various other embodiments. modifications. and equivalents thereof, which, after reading the description herein, may suggest themselves to those skilled in the art without departing from. the spirit of the present in ~entiora and/or the scope of thc appended claims.

10O94j Other features and advantages of the invention O it be apparent fn:)m the following descriptio-in of the preferred embodiments thereof and from the claims. These and many other variations and embodiments of the invention will be apparent to one of skill in the an capon a review of the appends description and examples.

EXAMPLE'S
EXAMPLE I

PRG4 mRNA Expression in Human Conical and Conjunctival Epithelial Cells [0100] Human curneaal epithelial cells were isolated from the coracoscleral rims of male and female donors. Cells were processed either directly (a = 8). or first cultured is phenol red-fee .keratinocvte serum :free media (n = 2). Bulbar conja.uac[iva.e (ra =
2). conjunctival aampress.ion cytology samples (aa = 9), immortalized hu.maaaa conjunnctmaal epithelial cells after culture (n 1). NOD mouse lacrimal laads (n adult raaice'sex 10 gltands/saa:aa.ple), and BALB/t mouse meihorn au glands (n --- 7 adult mice/sex, glands from l() 28 lads/sample) were obtained during surgical procedures. These samples were. processed for the analysis of PRG4 mRNA by using primaril , RT-PCR (n = IS human, all mouse) and Affymetrix GeneChips (n = 4 human corneas). The PRG4 primers for PCR
spanned over I .kbp of introit sequences, to order to suppress amplification of contaminating-chromosomal DNA (Table 1). Amplified samples were scree; ned for the presence of PRG4 14; Products, by usia:a arose gel electaophe rosis and. an Agilent 2100 Bioanaal zer. To confirm the identity of anaplieons. PCR pr ducts from cornea. samples (ra ---2), co.ajuncti cal epithelial cells (n W 1) and a human liver standard (a = 1) were sequenced with a 3100 Genetic Anal y.e:r at the Massachusetts Eye and Far Infinnary, DNA
Sequencing Center for Vision Research (Boston. MA) and resulting data were analyzed.
20 with Hl A.STn searches of GenBank dataihases.

Table .. Oligonucleotide primers designed for RT- PC R analysis of.PR64 mmmRN
Species Orientatioaa lracleot:ide sc.caraenc c: ( s 3) E:~ons Amplicon Size h,.
2 Human Sense GATf_GCAGGCGTA('{"Ã.C' AAA (SEQ.ID NO:2) 9-12 326 Anti.se:nse CAGACTTTGGA AAGG 1 C TGCC (S.EQ.I.I) N0:3) 101011 It was demonstrated that PRG4 raaRNA is present in all human conical and conjunctival epithelial cell and Impression cytology y stamples. The identity of PRG4 PCR.
30 products was confirmed by DNA sequence analysis (Fable 2). The results show thaat.PRG4 s transcribed in human corneal and connjunctiyaal epithelial cells, Table .2 Identification of amplicon sequence from human cornea, c~-,) junct \:al and liver samples Sequencing Aligned Base Pairs `total Base Pairs BLASTh Search Direction ToHrmanPRG4 fromAmplicon Idenntir' 1-lurman Liver Standard A l-or and 4535 00 Human PRG4 A Reverse. 488 491 Human PRG4 B Forward 496 499 1-human PRG4 B Reverse 498 500 Human PRG4 Human Cornea t24 year à ld :feanaa:le A Forward 497 499 H-human PRGI4 A Reverse 490 492 Human PRG4 B Forward 500 504 Hà in an PRG4 B Reverse 498 501 Human 11RG4 1-larnaan Cornea (51 year old female) 2]:] A Fors rd 498 499 1 lu man PRG4 RevaYrba;`. 474 489 Human PRG4 B Forward 4Ã96 498 Human PRG4 B Revs rsÃ; 490 491 Human PRG4 Hun= (onajun.cÃivaal LPih .l al Cells A Forward 496 499 Human PRG4 * Reverse 490 4922 Hu-nun PR64 B Forward 495 499 Human PRG4 B Reverse 474 491 Human 11RG4 Two different samples (A&- B) of each pz l.,rraatczzt'+ker se quCn ced:if]
forward w id reverse di eetions. 'l:lze human corm !,t s<tm Ales were el?itlieliaal cells from the cà n cl)" ieml rims of female d?za ?a,. ']']ac ,Z tk tt-c4õt to maul-te gear lamwul l'1Z0a iS NM t?0-W 7.

Regulation of PR64 E\presslon in vitro by Androgen 10102] Androgen treatment upregulaates PRG4 mRNA expression in primary human cortical epithelial cells. Methods. Cells Lucre: grown in keratinocvte serum free media until 5 reaching about l U% continence Cells (n - wellsf.re to ea lr`~. perIm t) were then incubated with either vehicle or 10 is vi dihxdrotcstostcrorae (DHT) for up to 5 days. At designated times cells were processed for total RNA isolation and analysis of mRNA ni.R.NA by RT- PCR. Results. The results show that the DHT-I' induces a marked increase in PR.G4 anRNA levels in priinar_y human cortical epithelial cells (figure 6). This 10 androgen ctf ct relative to the control levels at Day 0, became prominent after 3 (103-fold increase), 4 (3.6-fold iincrease) w id 5 (2. -fold increase) days of hormone exposure.
This DHT influence on PR64 mRNA expression in primary human cortical epithelial cells was confirmed in another experime.nt.. Treatment of cells for 5 days %N-with D1-1T caused a 46-f%_ ld increase in PRG4 rRNA content, relative to that of y ehiclt treated controls.

15 101031 Combined .17f3-estradio.1 and progesterone treatment downregulates PRt!'4 nmRNA
expression in mouse lacrimal tissue. Age-inaatched and young adult BAL. /c mice, that were ovariectomized when S weeks old, were purchased from Taconic laboratories (Germantown NY). .Animals were maintained in constant temperature rooms with fixed light, dark period of 12 hours duration. Ten days after surgea_ty,, pellets containing vehicle ?0 (thole st r )l. a:si.eÃli.~ l ellcsl os . lactose), or 17j3 stracliol (ll.5 a i g) plus progesterone (10 mg), were implanted subcutaneously in the o ~aieetonmiiz.c.d mice. The:
pellets y ver obtained from .innovative Research of America (Saratsotxa, Ft..) and were designed for the constant release of placebo or physiological W1otinis of Ste x steroid (:i.e.
as in pregnancy) for 3 weeks. After 14 days of treatment, mice (n ::: 7 mice:condition{
experuneiat) were 25 sacrificed by C02 inhalation. and exorbital lacrimal glands were removed, pooled according to group (n 14 glands/ ample) and processed for molecular biological procedures.

101041 Total RNA was extracted fl-0.111 tissues by using ".T'Ri.zol reagent (invitrogen Corp., Carlsbad, CA). When indicated,, samples were also exposed to RNasw-take DNase 30 (la itrogen), examined spc.cta photon triea l at 260 mini to de.tcria ine concentration and evaluated on 6,7% fc~n ,~lclcl~e de/i . `" agarose (C ihco/BRL Grand .Island, NY) gels to Verify RNA integrity The RNA samples were further purified with RN Aqueous spin columns (Anabion, Austin- Tx), and the integrity of these preparations was assessed with a RNA 6000 Nano LabChip with a" Agilent :7100 Rioanal zer (:AgIlent Technologies. Palo Alto, CA). The RNA samples were then processed for Code:I.:ink Bioarra-hybridization.
In brief, eDNA was synthesized from RNA. (2 Grg) w vitla a CodeLinl Expression Assay Reagent Kit (Ame.rslaaan. Piscataiwwav, NJ) and purified with a Ql quick purification kit (Qia en, Valencia, CA). After ampIc drying, eRNA w is n ide with a Codelin.k l xp:ress on .Assay Read ent Kit (Aniershaan), recovered ~,vith an RNÃe.asy kit (Qiagen) and quantifie with an UV spe.ctrophotoÃaaeter. Fraagmented, biotin-labeled cRNA
was then incubated and agitated (300 rpm shaker) on a Code-ink Bioarrda y at 37" C.:
for 18 hours.
The Bi:oaarray was waahed, exposed to streptav.idi.n Alexa 647- and scanned by using ScanArray Express soft are and a Scaaa_Arraay Express 14T scanner (Packard I
ioSrience, 11 e-ride-n. CT) w. pith the lasers ;t at 635 raa a. laser power at 100%Vi%%, and photornultiplier tube voltage at 60%. Scanned image files N ,,ere evaalaaatcd by utilising Codebnk image and data.
analysis software (Aanersham ), which produced both raw and. normalized . hN
bridiz:ation signal intensities for each array spot. The spot intensities (-10 ,000) on the raricroarray image a :era:- standardized to a median of 1. Normalized data, with signal intensities exceeding 0.75, were analyzed. with GeneSifter.Net software (Viz Labs L1.C, Seattle.
WA, vizxlabs.com). `statistical analysis of individual gene expression data was performed with Studerrt's t-test (two tailed. unpaired).

10105} The data showwww that combined esir d.iol and progesterone treatment, as compared to that of placcbo, causes a significant (p ::- 0.020), 1,6-old decrease in PRG4 gene expression in the mw-)use lacrirnal gland EXAMPLE .3 Treatment of Deficient Ocular Bounda - Lubrication in Vivo w:ww ith 1ndrogea and PRG4 101061 A patient complaining of ocular surface irritation is examined for ocular lubrication or conditions associated w with a deficiency in ocular lubrication by is easuri:ng symptoms greater than 2 positive responses on the Me onnies questionnaire, greater than 3e'_ a score of 5 on the Ocular Sur1aacc; Disease. Index (OSDI), or through c idence of some.

sympttonos on the Visual Analog Scale, in corrrbinrtion with objective signs including one or more of a reduced tear film breakup -time (less than = 10 seconds), inferior lateral tear aawraiscus osrr olarits' greater than 30S IIIOsarasrL:, low Schirmer strip value (less than = 10 MM)- sodium fluorescein conical or conjunctival staining (scorns 0 with multiple ntacropunctates), significant debris resulting, .from .impression cytology. n eibomian gland dysfunction however determitied, a decrease in the rate of frost-blink displacement of a contact lens, a change in the spatiotetupoaal transfer li c:tiota of a contact lens tbllowitag application of a series of pressure impulses, a decrease in the rate of post-blink inter fi rc attctric tear film relaxation, an increase in the concentration of proinflarrulr.atork.
cytokines. a reduced concentration oflactolferrin or lvsozvrn.c. or an increase in the rate of post-blink point spread function teen hc.rence.

101071 The patient administers I to 2 drops on the surface of each eve a solution containing 1õza-clilr Ala ratestosterone and 200 ltgimt, PGR4 protein suspended in an ophthalmically acceptable balanced salt solution. The patient is instructed to close their eves for 10 seconds.

101081 Follow-up visits may track a reduction in interior lateral tear osrnolarit . increased tear film breakup time, or the other aforementioned signs. In particular if the tear film r_onaolarity is reduced from an abnormal value (perhas 330 m srrts/L) to a more rion al value (perhaps 304 mOsmsi.L), the therapeutic modulation and replenishment of the ocular surface lubrication wvould be deemed successful, REFERENCES

1. G..1). Jay, Curr Capin Orthop l 5, 355 (2004).

2. Schumacher BL, Hughes CE, Kuettner KE, Caterson B. Asdelottc MB.
I:am munodetecuor and partial cDNA sequence of'tlhe protcogltiycan super f cial zone protean; synthesized by cells lining, s noviaal joints. J Orthop Res. 1999 Jan; 1.7(l):] 1Ã1-211.

3. S. G. Rees et al. Matrix Biology 21, 593 (2002) .

4. Schumacher III, Schmidt '1'A_ Voegtl me- MS_ Chen AC, Sala R.l-.
Proteoglycan 4 (PRG4) synthesis and :immunolocaliz<ation is bovine men:iscÃr . J Orthop Res.
2(105 May-?3,0062-8, . J. Marcelino Ã;t ;.al , Nat Genet 2' 7 319 0999).

O. D. K. Rhee et al.. J Clin Invest 115, Ã22(`2Ã}05).

7. Cutolo -" l_ Cape lino S, Sulli A. Se:noli B. Secchi ME. Vifl ag ;io B.
Straub RH.
l:st Fxcaas and a atoina a raa c diseases. Ann N Y Acad Sci 2Ã 06:1Ã 89:538-547.

S. Cutolo .3I Sull.i A. Capellino S_ Villaggio B.. Nlontagna P, IPizzorni C
I?acz.li:no S, Sc;riolo B, Felh L. Straub RH ..Anti-TNF and sex hormones. ,inn N -Ltd Sci 2Ã306;1Ã 69:391-400.

9. Rontzsch A. Thoss K, Petrm-w 1'I ..1-le n zyren S. Brauer R. Amelioration of'murinc antis + a -induced arthaitis by deh droepiandroste.:rone (DHl A). infla.n .m Res 2200$;53:189-19.

10. Schwarz lh . I-lills BA, Br. J. Rheum. 1998 3 7:21-20.

111 Jay GD, Hon BS. Connect Tissue Re, 1992; 280-2):89-98, 12, Jones MB. c t. al. Mathematical Medicine aand Biology 2005: 22.1265, 13. F \lcv r, 'R. M. Own o , K. Da nsf-e ki, T. Gti: aloy. Nanose.ià nce:
Friction and Rla oloo!v on the N,anonacter Scale. (World Suentiic Publishing Co. Pte,. Ltd, liver Fd-s 2_ Ne Jersey. 2002). pp. 373.

14. D. Dc ewwson, Proc l:nst Mech ErLt, 11-1j 215, 335 (2000.

15. G. A. Ateshian,, V. C. Mow, .in Basic Orthopaedic Biomechanics and Mechano-Bio1ogy V. C. Mow, R. Hw.tiskes. Fels. (Lippincott Willi im& Wilkins, Philadelphia.
2005) pp. 447-494.

16. F. Guilak, Arthritis Rheum 52.1Ã 3.2 (Jun. 2005).

17. K. C.Morel l, W W. A. Hodge, D. E. Krebs, R. W. Mann', Proc Natl Mad Sci U
S A 102.
14819 (Oct 11. 20)5).

1 S. S. '. Sz anson, in . clrilt A ocular Ca~rti age M. A. R. Freeman. Ed.
(Pitman 1tlc Baal- Tunbridge Wells. England, 1979) pp. 41?-460..

19. K. C. Morrelt. W. A. Hodge. D. F. Krebs, R. W. Mann. Proc Nat! Arad Sci U
S A 102, 14819 Oct 1.1.200. ).

20. C.. W. McC`ntehe:n, Fodd Proceedings 23õ 1061 (1916).

21. 'f. ;"~lurakami, Y. Sawa ;, M. Ilaara, JSMI_' Int 1 S vies C-NIechanica.1 Ss stcnis Ma hinn.e Elements &. Man i1'acturing 46, 594 (200.3).

22. G. Meach.in, Ann Rheum Dis 31.457 (1972).

23. Schmidt M. Nau a:n H. 'G reidle r C. , Schellenberg., M. Anders S. Siranb RH.
Inflammation. and sc lrou otu.. m :tabobsm..\,rm a \ ,cad Set 2(0tx:10 36 246 SEQUENCE LIST

SEQ ID \O: I
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P'I'-fPKI I'.'3P'T'-rPKk'T'AP'I, TPKCPTPTTPKEPAPTTKEPAPTTPkLPx\PT:'PKKPAPTTPKEP:r PTTPKEPPTTT'K
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GPSQTI-ITIRI.Q\'SPr' RLA\ QDKI.IVLHNEVK' SII.\\,'RC LP 'L'VTSA1SLPNIRKPDG
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SEQ ID NO2: :GATGCAGGGT:ACC;CCAAA (human. sense ) 0 SEQ ID NO.3. C ACGAC;TTTCIG tT:A.AGCGTCTGC"C (huuman. anit se ns ._)

Claims (24)

41, What is claimed is:

1. A pharmaceutical composition suitable for topical application to an ocular surface comprising a therapeutically effective concentration of a PRG4 inducing compound in combination with a therapeutically effective concentration of PRG4.

2. The pharmaceutical composition of claim 1, wherein the PRG4 inducing compound is selected from the group consisting of an androgen, an androgen analogue, a selective androgen receptor modulator, a selective estrogen receptor modulator. an estrogen antagonist, an aromatase inhibitor, an antiprotease a, proinflammatory cytokine antagonist, a cytokine release inhibitor, an antiinflammatory cytokine, an antiinflammatory agent a NF-.kappa.-B inhibitor, a proteasome inhibitor, and a combination thereof.

3. The pharmaceutical composition of claim 1, wherein the PRG4 inducing compound is in androgen or androgen analogue.

4. The pharmaceutical composition of claim 3, wherein the androgen analogue is selected from the group consisting of 17.alpha.-methyl-17.beta.-hydroxy-2-oxa-5.alpha.-androstan-3-one, testosterone, 4,5.alpha.-dihidrostestosterone, 17.beta.-hydroxy-5.alpha.-androstane containing a ring A. unsaturation. 19-nortestosterone, nitrogen-substituted androgen, and derivatives thereof.

5. The pharmaceutical composition of claim 2, wherein the selective androgen receptor modulator is selected from the group consisting of an aryl-propionamide compound, a bicyclic hydantoin analogue, a tetrahydroquinoline analogue, and a quinoline analogue.

6. The pharmaceutical composition of claim 5, wherein the ary-1-propionamide compound is S-3-(acetylamino-phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3- or trifluoromethyl-phenyl)-propionamide [S4], or S-3-(fluorophenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethyl-phenyl)-propionamide [S-1]).

7. The pharmaceutical composition of claim 2, wherein the proinflammatory cytokine antagonist is selected from the group consisting of an anti-TNF.alpha.
antibody, a soluble, TNF.alpha. receptor, and an IL-1 receptor antagonist.

8. The pharmaceutical composition of claim 21, wherein the antiinflammatory cytokine is TGF-.beta..

9. The pharmaceutical composition of claim 2, wherein the antiinflammatory agent is selected from the group consisting of cyclosporine A, c-Jun N-terminal kinase (JNK) inhibitor; extracellular-signal regulated kinase (ERK) inhibitor mitogen-activated protein (MAP) kinase inhibitor, matrix metalloproteinase (MMP) inhibitor, omega 3 fatty acid, and omega 6 fatty acid.

10. The pharmaceutical composition of claim 1,wherein PRG4 inducing compound is in the therapeutically effective concentration of 0.0001-0.1% w/v.

11. The Pharmaceutical composition of claim 1, wherein PRG4 is in the therapeutically effective concentration of 1.0- 10,000 µg/mL.

12. The pharmaceutical composition of claim 1, wherein PRG4 is in the therapeutically effective concentration of 50-500 µg/mL.

13. The pharmaceutical composition of claim 1, further comprising sodium hyaluronate or hyaluronic acid in a therapeutically effective of 10-100,1000 ug/mL.

14. The pharmaceutical composition of claim 13, wherein sodium hyaluronate or hyaluronic acid is in a therapeutically effective concentration of 500-5,000 µg/mL.

15. The pharmaceutical composition of claim 1, further comprising one or more surface active phospholipids selected from the group consisting of L-.alpha.-dipalmitoylphosphatidycholine, phosphatidylcholine, phosphatidylethanolamine and sphingmyelin in a therapeutically effective effective concentration of 10-10,000 µg/mL.

16. The pharmaceutical composition of claim 1, further comprising a phosphate buffered saline solution or an opthalmically acceptable balanced salt solution comprising a therapeutically effective concentration of one or more electrolytes selected from the group consisting of sodium phosphate, sodium chloride, potassium chloride, sodium bicarbonate, potassium bicarbonate, calcium chloride, magnesium chloride, trisodium citrate.
hydrochloric acid, and sodium hydroxide.

17. The pharmaceutical composition of claim 1, further comprising a therapeutically effective concentration of one or more ophthalmic demulcents, excipients, astringents, vasoconstrictors, and emollients.

18. The pharmaceutical composition of claim 1, further comprising a residence-time increasing agent.

19. The pharmaceutical composition of claim 1, wherein the PRG4 has an average molar mass of between 50 kDa and 400 KDa and is either recombinant or naturally occurring.

20. A method fur treating ocular lubrication deficiency, or symptoms associated therewith, in an individual in need thereof comprising topically administering to the ocular surface of the individual in need an effective amount of the pharmaceutical composition of any of claims 1-19.

21. The method of claim 20, wherein the condition is aqueous or evorative dry eye disease, Sjögrens syndrome keratoconjunctivitis sicca, androgen deficiency, meibomian gland disease, estrogen replacement therapy, contact lens wear, refractive surgery, allergy, reduced tear film breakup time, compromised tear film allergy, ocular surface disorders, increased protease levels in the tear film and at the ocular surface, chronic inflammation, hyperosmolarity, aging, and combinations thereof.

22. A method fur treating a condition that is associated with or causes a deficiency in ocular lubrication, or the symptoms thereof, in an individual in need thereof comprising topically administering to the ocular surface an effective amount of the pharmaceutical composition of any of claims 1-19.

21 The method of claim 22, wherein the condition is aqueous or evaporative dry eye disease, Sjögren's syndrome, keratoconjunctivitis, sicca, androgen deficiency, meibomian gland disease estrogen replacement therapy, contact lens wear, refractive surgery, allergy, reduced tear film breakup time, compromised tear film, allergy, ocular surface, disorders, increased protease levels in the tear film and at the ocular surface, chronic inflammation, hyperosmolarity, aging, and combinations thereof.

24. A method of locally inducing PRG4 on an ocular surface comprising topically administering to the ocular surface of an individual in need thereof a pharmaceutical composition of any of claims 1-19.