Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
SMALL ORGANIC MOLECULES THAT INCREASE THE ACTIVITY OF GELATINASE A IN OCULAR CELLS
Document Type and Number:
WIPO Patent Application WO/2004/028538
Kind Code:
A1
Abstract:
Small organic molecules capable of effecting a 'pharmacological trabeculocanalotomy' in an eye by means of reducing juxtacanalicular meshwork as a barrier to outflow of aqueous. The small organic molecules increase Gelatinase A activity in ocular cells by increasing cell membrane expression of membrane-type matrix metalloproteinases (MT-MMPs) to increase aqueous outflow as a treatment for primary open angle glaucoma.

Inventors:
Pfeffer, Bruce Alan (8 Creek Bend Drive, Fairport, NY, 14450, US)
Flick, Rosemarie Beth (3 North Emerson Avenue, Copiaque, NY, 11726, US)
Shams, Naveed Bin Kamal (11215 Crofton Overlook Court, Duluth, GA, 30097, US)
Bartels, Stephen Paul (16 Silco Hill, Pittsford, NY, 14534, US)
Application Number:
PCT/US2003/029528
Publication Date:
April 08, 2004
Filing Date:
September 19, 2003
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
BAUSCH & LOMB INCORPORATED (One Bausch & Lomb Place, Rochester, NY, 14604-2701, US)
International Classes:
A61K31/4453; C07C275/10; C07C281/18; C07D223/26; C07D279/22; C07D295/205; (IPC1-7): A61K31/4453; C07D211/16; C07D223/22; C07D279/22
Foreign References:
EP0035945A2
FR1455835A
US3686409A
US3975533A
GB1365847A
Other References:
AMBROSIO A F ET AL: "MECHANISMS OF ACTION OF CARBAMAZEPINE AND ITS DERIVATIVES, OXCARBAZEPINE, BIA 2-093, AND BIA 2-024" NEUROCHEMICAL RESEARCH, PLENUM PRESS, NEW YORK, US, vol. 27, no. 1/2, February 2002 (2002-02), pages 121-130, XP009022844 ISSN: 0364-3190
DATABASE CROSSFIRE BEILSTEIN BEILSTEIN INSTITUT ZUR FOERDERUNG DER CHEMISCHEN WISSENSCHAFT,FRANKFURT AM MAIN, DE; XP002267330 Database accession no. BRN: 6881299 & FARMACO ED. SCI., vol. 38, no. 4, 1983, pages 255-264,
HEWITT R E ET AL: "The activation, expression and function of gelatinase A (MMP-2)" TRENDS IN GLYCOSCIENCE AND GLYCOTECHNOLOGY, FUJISHIRO, JP, vol. 8, no. 39, January 1996 (1996-01), pages 23-36, XP002948336 ISSN: 0915-7352
LI L ET AL: "Protein tyrosine phosphorylation in signalling pathways leading to the activation of gelatinase A: activation of gelatinase A by treatment with the protein tyrosine phosphatase inhibitor sodium orthovanadate" BIOCHIMICA ET BIOPHYSICA ACTA. MOLECULAR CELL RESEARCH, ELSEVIER SCIENCE PUBLISHERS, AMSTERDAM, NL, vol. 1405, no. 1-3, 21 October 1998 (1998-10-21), pages 110-120, XP004277862 ISSN: 0167-4889
Attorney, Agent or Firm:
Vacca, Rita D. (Bausch & Lomb Incorporated, One Bausch & Lomb Place Rochester, NY, 14604-2701, US)
Lavoie, Paul T. (Bausch & Lomb Incorporated, One Bausch & Lomb Place Rochester, NY, 14604, US)
Download PDF:
Claims:
We claim :
1. Small organic molecules useful to increase the activity of Gelatinase A in ocular cells comprising: wherein W is selected from the group consisting of oxygen, sulfur and C 20 hydroxylated carbon group; Z is selected from the group consisting of oxygen and sulfur ; Ri is branched or unbranched and substituted or unsubstituted selected from the group consisting of C625 aryl, C320 cycloalkyl, C320 cycloalkenyl, C330 heterocyclyl, C630 heteroaryl, C630 aryloxy, C630 arylalkoxy, C630 aryloxycarbonyl, C630 arylaminocarbonyl, C630 arylalkyl, C630 aryloyl, C630 arylalkylcarbonyl, C630 arylalkoxycarbonyl, C630 arylthioalkyl, C630 arylalkenyl, C630 arylalkynyl, C630 aryloxyalkyl, C630 arylalkylthioalkyl, C630 arylalkoxyalkyl, C630 arylthiocarbonyl, C630 arylalkylaminocarbonyl, C630 NalkylN arylaminocarbonyl, Ceso Narylaminosulfonyl, Ceso Narylaminosulfinyl, C630 <BR> <BR> <BR> <BR> arylsulfonyl, C630 arylsulfinyl, C630 aryloyloxy, C630 arylthio, C630 arylalkylthio, C6 30 arylacyloxy, C63o arylalkylsulfonyl, C630 arylalkylsulfinyl, C630 <BR> <BR> <BR> <BR> arylalkylthiocarbonyl, C630 NalkylNarylaminosulfonyl, C630 NalkylN<BR> <BR> <BR> <BR> <BR> arylaminosulfinyl, C630 aryloxycarbonylalkylcarbonyl, C630 aryloylthio and C630 arylacylthio, and Ri as defined above when substituted is substituted with one or more substituents selected from the group consisting of C125 alkyl, C125 alkenyl, C125 alkynyl, C125 alkoxy, C125 alkenoxy, hydroxy, carboxy, amino, C125 (N alkylcarbonyl) amino, C125 (Nalkylcarbonyl)Nalkylamino, C125 (N alkylcarbonylalkyl) amino, cyano, nitro, C630 arylazo, sulfo, sulfino, sulfhydryl, halo, C125 haloalkyl, trifluoromethyl, C125 trifluoromethylalkyl, C630 arylalkyl, C630 aryl, C125 Nalkylamino, C125 Ndialkylamino, C125 (NalkylNalkenyl) amino, C1 25 Ndialkenylamino, C125 alkylsulfonyl, C125 alkylsulfinyl, C125 alkylthio, hydrido, C125 cyanoalkyl, C125 acyl, C125 alkylcarbonyloxy, nitroso, C125 alkoxyalkyl, C125 alkoxycarbonyl, C125 hydroxyalkyl, thiocarboxy, C125 thiocarboxyalkyl, C125 alkylthiocarbonyl, C125 alkylthiocarbonylalkyl, C125 alkoxythiocarbonyl, C125 alkoxythiocarbonylalkyl, sulfamoyl, sulfinamoyl, C125 Nalkylsulfamoyl, C125 N dialkylsulfamoyl, C125 Nalkylsulfinamoyl, C125 Ndialkylsulfinamoyl, C125 sulfamoylalkyl, C125 sulfinamoylalkyl, aminocarbonyl, C125 aminocarbonylalkyl <BR> <BR> <BR> <BR> aminocarbonylmethyl, C125 Nalkylaminocarbonyl, C125 Ndialkylaminocarbonyl, C125 alkoxycarbonylamino, thiocarbamoyl, C125 thiocarbamoylalkyl, C125 (N alkyl) thiocarbamoyl, C125 (Ndialkyl) thiocarbamoyl, aminothio, C125 alkylaminothio, C125 Ndialkylaminothio, C125 cycloalkyl, C125 cycloalkenyl, C125 aryloxy, C125 cycloalkoxy, C125 cycloalkenoxy, heterocycloxy, C125 heteroaryloxy, C125 heteroaryl, heterocyclic amines, heterocyclic amides, Nor Soxo and Nor Sthioxo; R2 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C110 alkyl, C110 alkenyl, C110 alkynyl, cyano, C110 haloalkyl, trifluoromethyl, C615 cycloalkyl, C615 cycloalkenyl, C615 heterocyclyl, C630 aryl, C630 heteroaryl, halo, nitro, C630 alkylaryl, C630 alkoxyaryl, C115 alkylcarbonyl, hydroxycarbonyl, aminocarbonyl, C115 N alkylaminocarbonyl and C115 alkoxycarbonyl, and R2 as defined above may be substituted with one or more substituents as defined above for Ri ; R3 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C112 alkyl, C112 alkenyl, C112 alkynyl, C120 alkoxyalkyl, C120 alkoxyalkenyl, C120 alkenoxyalkyl and C120 alkenoxyalkenyl, and R3 as defined above may be substituted with one or more substituents as defined above for Ri ; and R4 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C112 alkyl, C112 alkenyl, C112 alkynyl, C120 <BR> <BR> alkoxyalkyl, C120 alkoxyalkenyl, C120 alkenoxyalkyl, C120 alkenoxyalkenyl, C630 aryl, C630 cycloalkyl, C630 cycloalkenyl, C630 heterocyclyl and C630 heteroaryl, and R4 as defined above may be substituted with one or more substituents as defined above for Ri.
2. The small organic molecules of claim 1 wherein Ri is selected from the group consisting of phenyl, benzoyl, 2, 6difluorophenyl, 4 trifluoromethylphenyl, 3, 4dimethoxyphenyl, 4tertbutylphenyl, 4methylphenyl, 4methoxyphenyl, 4phenoxyphenyl and 2thienyl.
3. The small organic molecules of claim 1 wherein R2 is selected from the group consisting of cyano, cyclopentanyl, phenyl and trifluoromethyl.
4. The small organic molecules of claim 1 wherein R3 is selected from the group consisting of hydrido and alkyl.
5. The small organic molecules of claim 1 wherein R4 is selected from the group consisting of phenyl, cyclohexyl and alkyl.
6. The small organic molecules of claim 1 wherein R3 and R4 are fused in a heterocyclic ring.
7. The small organic molecules of claim 1 wherein R3 and R4 are fused in a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, 4 methylpiperidino and morpholino.
8. Small organic molecules useful to increase the activity of Gelatinase A in ocular cells comprising: and.
9. A small organic molecule useful to increase the activity of Gelatinase A in ocular cells selected from the group consisting of adenylyl cyclase inhibitors, phospholipase D activators, cyclic adenosine monophosphate phosphodiesterase activators, protein kinase A inhibitors, protein phosphatase inhibitors, molybdat oxoanions, tungstate oxoanions and dephostatins, phosphatidate phosphohydrolase inhibitors/cationic amphiphiles, Rho activators, lipid lowering agents or hyperlipoproteinemics and concanavalin A receptor ligands.
10. A small organic molecule useful to increase the activity of Gelatinase A in ocular cells selected from the group consisting of 9 (tetrahydro2'furyl) adenine, 2', 5'dideoxyadenosine, miconazole, roxithromycin, fluoride ion, bradykinin, progesterone, endothelin, vasopressin, 4 hydroxynonenal, interleukin11, angiotensin 11, N (6aminohexyl)5chloro1 naphthalenesulfonamide, oxidized low density lipoprotein, phosphatidic acids, dioleoyl, dioctanoyl, 1stearoyl2arachidonylsnglycerol3phosphate, phosphatidic acid analogues, thiophosphatidic acid, phenylalanine, pyrazinyl guanidine, (N[2((pbromocinnamyl) amino) ethyl]5isoquinolinesulfonamide, vanadium salts, potassium bisperoxo (1, 10phenanthroline) oxovanadate, dipotassium bisperoxo (picolinate) oxovanadate, molybdate oxoanions, tungstate oxoanions, dephostatins, 3, 4dihydroxyNmethylNnitrosoaniline, 3,6dihydroxy NmethylNnitrosoaniline, propranolol, tetracaine, mepacrine, desmethylimipramine, chlorpromazine, desipramine, sphingosine1phosphate, 2tetradecylglycidic acid, 5 (tetradecyloxy)2furoic acid, 3thiadicarboxylic acid, 3 (4methylpiperazin1yl)1phenylpropanone, 6,7dihydro5H dibenz [c, e] azepine, N2nbutylindazolone, 4phenyl5, 5 dicarbethoxy2pyrrolidinone, 4 (4hydroxy3iodophenoxy) 3,5 diiodohydrocinnamic acid, 1methyl4piperidyl bis (pchlorophenoxy) acetate, 2 [ [1methyl2 [3 (trifluoromethyl) phenyl] ethyl] amino] ethanol benzoate ester, 5 methylpyrazinecarboxylic acid 4oxide, acetylcholinesterase and complement protein 1q.
11. Small organic molecules useful to increase the activity of Gelatinase A in ocular cells comprising: wherein R6 is branched or unbranched and substituted or unsubstituted selected from the group consisting of C625 aryl, C320 cycloalkyl, C320 cycloalkenyl, C330 heterocyclyl, C630 heteroaryl, C630 arylalkyl, C630 arylthioalkyl, C630 arylalkenyl, C630 arylalkynyl, C630 aryloxyalkyl, C630 arylalkylthioalkyl and C630 arylalkoxyalkyl, and R6 as defined above when substituted is substituted with one or more substituents selected from the group consisting of hydroxy, carboxy, amino, cyano, nitro, nitrate, C125 alkyl, C125 alkenyl, C125 alkynyl, C125 alkoxy, C125 alkenoxy, hydroxy, carboxy, amino, C125 (Nalkylcarbonyl) amino, C125 (N alkylcarbonyl)Nalkylamino, C125 (Nalkylcarbonylalkyl) amino, C630 arylazo, sulfo, sulfino, sulfhydryl, halo, C125 haloalkyl, C125 trifluoromethylalkyl, C630 arylalkyl, C630 aryl, C125 Nalkylamino, C125 Ndialkylamino, C125 (NalkylN alkenyl) amino, C125 Ndialkenylamino, C125 alkylsulfonyl, C125 alkylsulfinyl, C125 alkylthio, hydrido, C125 cyanoalkyl, C125 acyl, C125 alkylcarbonyloxy, nitroso, C 25 alkoxyalkyl, C125 alkoxycarbonyl, C125 hydroxyalkyl, thiocarboxy, C125 thiocarboxyalkyl, C125 alkylthiocarbonyl, C125 alkylthiocarbonylalkyl, C125 alkoxythiocarbonyl, C125 alkoxythiocarbonylalkyl, sulfamoyl, sulfinamoyl, C125 N alkylsulfamoyl, C125 Ndialkylsulfamoyl, C125 Nalkylsulfinamoyl, C125 Ndialkylsulfinamoyl, C25 sulfamoylalkyl, C125 sulfinamoylalkyl, aminocarbonyl, C125 aminocarbonylalkyl, C125 N alkylaminocarbonyl, C125 Ndialkylaminocarbonyl, C125 alkoxycarbonylamino, thiocarbamoyl, C125 thiocarbamoylalkyl, C125 (Nalkyl)thiocarbamoyl, C125 (N dialkyl) thiocarbamoyl, aminothio, C125 alkylaminothio, C125 Ndialkylaminothio, C125 cycloalkyl, C125 cycloalkenyl, C125 aryloxy, C125 cycloalkoxy, C125 cycloalkenoxy, heterocycloxy, C125 heteroaryloxy, C125 heteroaryl, heterocyclic amines, heterocyclic amides, Nor Soxo and Nor Sthioxo; Y is selected from the group consisting of O and S; the R7 groups may be the same or different selected from the group consisting of H or C115 alkyl ; R8 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, trifluoromethyl, cyano, C112 alkyl, C112 alkenyl, C112 alkynyl, C125 alkoxy, C125 alkenoxy, C125 alkynoxy, C125 alkylthio, C125 alkylcarboxy, C125 alkenylcarboxy, C125 alkynylcarboxy, C125 aminoalkyl, C125 alkylaminoalkyl, C125 dialkylaminoalkyl, C630 aryl, C630 cycloalkyl, C630 cycloalkenyl, C630 heterocyclyl, C630 heteroaryl, C630 arylalkyl, C630 cycloalkylalkyl, C630 cycloalkenylalkyl, C630 cycloalkylcarboxy, C630 cycloalkenylcarboxy, C630 heterocyclyalkyl, C630 heteroarylalkyl, C630 arylcarboxy, Ceso heterocyclylcarboxy, and C630 heteroarylcarboxy, and R8 as defined above may be substituted with one or more substituents as defined above for R6 ; R9 is either Rio is selected from the group consisting of O, S, CHNO2, NNO2, NS (0) 2CH3 and NCN; Q is NR8R8 m is an integer less than 3; and n is an integer less than 7.
12. The organic molecule of claim 11 wherein R6 is naphthyl or 2,3 dichlorophenyl, Y is oxygen, both R7 groups are hydrido, n is two or three, R8 is isopropyl, R9 is Rio is oxygen and Q is NR$R$ with one R8 group hydrido and the other hydrido or methyl.
13. Small organic molecules useful to increase the activity of Gelatinase A in ocular cells comprising: wherein R6 is branched or unbranched and substituted or unsubstituted selected from the group consisting of C625 aryl, C320 cycloalkyl, C320 cycloalkenyl, C330 heterocyclyl, c630 heteroaryl, C630 arylalkyl, C630 arylthioalkyl, C630 arylalkenyl, C630 arylalkynyl, C630 aryloxyalkyl, C630 arylalkylthioalkyl and C630 arylalkoxyalkyl, and R6 when substituted is substituted with one or more substituents selected from the group consisting of hydroxy, carboxy, amino, cyano, nitro, nitrate, C125 alkyl, C125 alkenyl, C125 alkynyl, C125 alkoxy, C125 alkenoxy, hydroxy, carboxy, amino, C125 (Nalkylcarbonyl) amino, C125 (N alkylcarbonyl)Nalkylamino, C125 (N alkylcarbonylalkyl) amino, C630 arylazo, sulfo, sulfino, sulfhydryl, halo, C125 haloalkyl, trifluoromethyl, C125 trifluoromethylalkyl, C630 arylalkyl, C630 aryl, C125 Nalkylamino, C125 N dialkylamino, C125 (NalkylNalkenyl) amino, C125 Ndialkenylamino, C125 alkylsulfonyl, C125 alkylsulfinyl, C125 alkylthio, hydrido, C125 cyanoalkyl, C125 acyl, C125 alkylcarbonyloxy, nitroso, C125 alkoxyalkyl, C125 alkoxycarbonyl, C125 hydroxyalkyl, thiocarboxy, C125 thiocarboxyalkyl, C125 alkylthiocarbonyl, C125 alkylthiocarbonylalkyl, C125 alkoxythiocarbonyl, C125 alkoxythiocarbonylalkyl, sulfamoyl, sulfinamoyl, C125 Nalkylsulfamoyl, C125 Ndialkylsulfamoyl, C125 Nalkylsulfinamoyl, C125 Ndialkylsulfinamoyl, C125 sulfamoylalkyl, C125 sulfinamoylalkyl, aminocarbonyl, C125 aminocarbonylalkyl, C125 N alkylaminocarbonyl, C125 Ndialkylaminocarbonyl, C125 alkoxycarbonylamino, thiocarbamoyl, C125 thiocarbamoylalkyl, C125 (Nalkyl)thiocarbamoyl, C125 (N dialkyl) thiocarbamoyl, aminothio, C125 alkylaminothio, C125 Ndialkylaminothio, Ci25 cycloalkyl, C125 cycloalkenyl, C125, C125 cycloalkoxy, C125 cycloalkenoxy, heterocycloxy, C125 heteroaryloxy, C125 heteroaryl, heterocyclic amines, heterocyclic amides, Nor Soxo and Nor Sthioxo; Y is selected from the group consisting of O and S; R11 is selected from the group consisting of hydrido, C115 branched or straight alkyl and CI15 branched or straight alkenyl ; R12 is selected from the group consisting of hydrido and C115 alkyl ; L is selected from the group consisting of nitrate, YR13 and NRsRe ; R13 is selected from the group consisting of CI15 alkyl, C115 alkenyl, C115 alkynyl, C115 haloalkyl, trifluoromethyl and cyanomethyl ; R8 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, trifluoromethyl, cyano, C112 alkyl, C112 alkenyl, C112 alkynyl, C125 alkoxy, C125 alkenoxy, C125 alkynoxy, C125 alkylthio, C125 alkylcarboxy, C125 alkenylcarboxy, C125 alkynylcarboxy, C125 aminoalkyl, C125 alkylaminoalkyl, C125 dialkylaminoalkyl, C630 aryl, C630 cycloalkyl, C630 cycloalkenyl, C630 heterocyclyl, C630 heteroaryl, C630 arylalkyl, C630 cycloalkylalkyl, C630 cycloalkenylalkyl, C630 cycloalkylcarboxy, C630 cycloalkenylcarboxy, C630 heterocyclyalkyl, C630 heteroarylalkyl, C630 arylcarboxy, C630 heterocyclylcarboxy, and C630 heteroarylcarboxy, and R8 as defined above may be substituted with one or more substituents as defined above for R6 ; Rg is either Rio is selected from the group consisting of O, S, CHNO2, NN02, NS (0) 2CH3 and NCN; Q is NR8R8 ; m is an integer less than 3; q is an integer less than 5; and t is an integer less than 9.
14. Small organic molecules useful to increase activity of Gelatinase A in ocular cells selected from the group consisting of piperidyl carbonyl nitrile oxime molecules, aryloxydialkylurea molecules, guanabenz molecules and tricyclic molecules.
15. A pharmaceutically acceptable composition comprising: one or more small organic molecules or one or more hydrates of the molecules in a therapeutical effective amount to increase the activity of Gelatinase A in ocular cells.
16. A pharmaceutical acceptable composition comprising: one or more small organic molecules or one or more acid addition salts of the molecules in a therapeutical effective amount to increase the activity of Gelatinase A in ocular cells.
17. A pharmaceutical acceptable composition comprising: one or more small organic molecules or one or more base addition salts of the molecules in a therapeutically effective amount to increase the activity of Gelatinase A in ocular cells.
18. A method of administering the pharmaceutically acceptable composition of claim 15,16, or 17 comprising: formulating said composition as a sterile aqueous or nonaqueous solution; and applying said solution on or within an eye.
19. A method of administering the pharmaceutical acceptable composition of claim 15,16, or 17 comprising: providing said composition in the form of an ocular implant ; and implanting said ocular implant within an eye.
Description:
SMALL ORGANIC MOLECULES THAT INCREASE THE ACTIVITY OF GELATINASE A IN OCULAR CELLS Field of the Invention : The present invention relates generally to small organic molecules capable of effecting a"pharmacologic trabeculocanalotomy"in an eye by means of reducing juxtacanalicular meshwork as a barrier to outflow of aqueous. More specifically, the present invention relates to small organic molecules that increase Gelatinase A activity in ocular cells by increasing cell membrane expression of membrane-type matrix metalloproteinases (MT-MMPs) to increase aqueous outflow as a treatment for primary open angle glaucoma. The present invention likewise includes methods of manufacturing and using such small organic molecules in the treatment of primary open angle glaucoma.

Background of the Invention : The extracellular matrix (ECM) of an eye is an association of specialized proteins, glycoproteins, and proteoglycans, that subserve and impart structure to the physiological functions of connective tissues. At the cellular level, the ECM not only provides structure, flexibility and support, but also acts as a filtration barrier, mediates cell attachment and influences tissue morphogenesis and differentiation. Part of the normal functioning of the ECM involves the ECM's tightly regulated turnover, which balances the degradation and disposal of effete molecules with the secretion and integration of the various newly synthesized ECM elements.

Specialized extracellular proteolytic enzymes, termed matrix metalloproteinases (MMPs), are produced by many cell types. MMPs play an important role in the initial degradation of such ECM molecules as collagen, fibronectin and various proteoglycans. MMP activity is regulated in part through secretion as inactive proenzymes and activation by proteolytic processing to smaller molecular weight forms. This regulated activity of MMPs usually requires protease activity as well as autolytic mechanisms. MMPs are inhibited by endogenous tissue inhibitors of matrix metalloproteinases (TIMPs).

For the MMP, Gelatinase A (GeIA) (MMP-2; 72 kD gelatinase ; type IV collagenase ; E. C. 3.4. 24.24), the specific proteolytic activator is known to be another member of the MMP family, namely MT-MMP. In contrast to other MMPs, MT-MMP is predominantly expressed as an integral membrane protein.

There are six known subtypes of MT-MMP, hence the designations MT1-MMP for MMP-14, MT2-MMP for MMP-15, MT3-MMP for MMP-16, MT4-MMP for MMP-17, MT5-MMP for MMP-24 and MT6-MMP for MMP-25. All subtypes of MT-MMP, except for MT4-MMP, effect a cleavage in 72 kD proGelA to initiate a proteolytic"cascade"to 66 kD (intermediate), 59 kD (active) and 43 kD ("mini") forms of GelA. MT-MMP is capable of activating GelA that is complexed with a specific inhibitory protein, TIMP-2. This places MT-MMP expression and/or activity as a major control point in the regulation of ECM turnover.

Trabecular meshwork (TM) is the tissue located at the irido-corneal angle of an eye's anterior segment. The TM is where the aqueous secreted by the ciliary epithelium flows out of the eye. The cells of the TM reside either on collagenous beams, or trabeculae, or embedded in the ECM associated with the canal of Schlemm. The canal of Schlemm is an endothelium-lined channel into which the aqueous drains. The intraocular fluid pressure (IOP) is maintained through a balance of the secretion and outflow of aqueous. Normal IOP is slightly above venous pressure, in part resulting from outflow resistance at the TM. TM outflow resistance is believed to be the result of the hydrodynamic properties of the ECM macromolecules of the TM and the ECM associated with the trabeculae.

For the complex of potentially blinding eye diseases termed primary open angle glaucoma (POAG) the hallmark of which is an insidious, progressive increase in IOP, a prevailing theory of the etiology of the disease is a dysfunction in the regulation of ECM turnover at the level of the TM. There is a biochemical lesion localized to the TM, which manifests as a general excess of ECM or as an imbalance with respect to specific components of the ECM, either of which impairs the ability of fluid to leave the eye at its normal physiological rate. It has been proposed that pharmacological intervention to reduce accumulated ECM could result in a lowering of the elevated IOP characteristic of these diseases.

A therapeutic small organic molecule capable of increasing MT- MMP expression has been decribed by Ito et al. (Ito et al., Eur. J. Biochem. 251, 353-358 (1998) ). As described by Ito et al., a trifluoperazine treatment of human cervical fibroblasts resulted in MT1-MMP-induced activation of Gelatinase A (GeIA). Trifluoperazine had previously been categorized as a therapeutic "antipsychotic."Ito et al. however, classified trifluoperazine as a calmodulin antagonist, and made a similar claim for another calmodulin inhibitor, W-7, although the effect from the later compound was not particularly pronounced. Ito et al. deduced that calmodulin negatively regulates MT-MMP expression.

Using Western blot immunochemistry, the presence of MT-MMP has been documented in human ocular tissues other than TM cells, as well as in fresh and cultured porcine TM cells. (Alexander, J. and Acott, T. S., Invest.

Ophthalmol. Vis. Sci. 40 (ARVO Abstracts): S506, #2670 (1999) ). (Smine, A. and Planter, J. J. , Curr. Eye Res. 16: 925 (1997)). In neither case was activation of GelA documented, although Alexander and Acott described increased expression of MT1-MMP with phorbol ester (phorbol 12-myristate 13-acetate).

Phorbol ester, however, does not have therapeutic usefulness since it is a known carcinogen.

In addition to trifluoperazine and phorbol ester, the following agents have been shown to increase MT-MMP expression and/or GelA activation in cells other than those of the TM.

Concanavalin A Interleukin-1 a Orthovanadate A Hexapeptide derived from Elastin Cytochalasin D Monensin Tumor Necrosis Factor-alpha.

Bacterial Lipopolysaccharide Hydrogen Peroxide Oxidized Low Density Lipoproteins Hepatocyte Growth Factor/Scatter Factor beta-Amyloid Peptide Activated Protein C Growth Hormone Interleukin VIII Glycyl-L-Histidyl-L-Lysine-Cu 2+ Lysophosphatidic Acid A search of U. S. patents found only one patent, U. S. Patent Number 5,260, 059, covering agents that increase the activity of matrix metalloproteinases (MMPs). The'059 patent discloses a method of treating glaucoma by providing TM cells with an array of macromolecules, including matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-3 (MMP-3). The class of MMPs designated as MT- MMPs had not been characterized at the time the'059 patent was filed. At the time of filing the'059 patent, physiologic activation of GelA was suspected of being brought about by means of autocatalytic mechanisms alone. Other molecules specifically mentioned in the'059 patent are basic heparin-binding growth factor, nerve growth factor, interleukin-1, interleukin-6, phorbol ester, calcium ions, zinc ions, plasmin, trypsin, and aminophenyl mercuric acetate (APMA).

Accordingly, a need still exists in the art to provide compositions and methods for the treatment of diseases termed primary open angle glaucoma.

Summary of the Invention : Small organic molecules in accordance with the present invention are therapeutically useful in the treatment of diseases termed primary open angle glaucoma by having a pharmacological effect on cells and tissue. The subject small organic molecules increase the expression or enzymatic activity of MT- MMP, or a similar enzyme expressed in the TM, that activates GelA. Activation of GelA leads to increased degradation of ECM and a subsequent increase in aqueous outflow with a resultant decrease in IOP.

The small organic molecules of the present invention are also useful in establishing model systems for finding new drug therapies for diseases such as primary open angle glaucoma. To accomplish the same, the subject small organic molecules are used to modulate TM cells and tissues in vitro to effect increased expression and/or activity of MT-MMP. Evidence for this would be the production of active species of GelA from proGelA. The activation of GelA may be demonstrated utilizing proGelA either secreted endogenously by TM cells, or added exogenously as a purified enzyme to experimental tissue or cells. In this respect it is important to note that aqueous humor has abundant proGelA. Enhancement of MT-MMP levels or activity in the cells of the outflow system of the eye would make GelA from proGelA locally available for proteolytic remodelling of ECM.

Given the importance of ECM regulation throughout the tissues and organs of the body, it is not surprising that many diseases have, as part of their pathology, an association with excessive degradation of ECM as a result of increased MMP activity. Drugs to inhibit MMP activity would be useful as therapies for diseases associated with the apparent loss of normal function and regulation of MMP. The present invention therefor includes therapeutic methods of treating particular diseases through cellular regulation of MT-MMP activity and/or expression using the small organic molecules of the present invention.

Accordingly, it is an object of the present invention to provide small organic molecules effective in the treatment of primary open angle glaucoma.

Another object of the present invention is to provide small organic molecules effective in the cellular regulation of MT-MMP activity and/or expression.

Another object of the present invention is to provide a method of effectively treating primary open angle glaucoma.

Another object of the present invention is to provide a method of effectively treating particular diseases through cellular regulation of MT-MMP activity and/or expression.

Another object of the present invention is to provide a method of manufacturing small organic molecules effective in the treatment of particular diseases through cellular regulation of MT-MMP activity and/or expression.

Still another object of the present invention is to provide a method of manufacturing small organic molecules effective in the treatment of primary open angle glaucoma.

These and other objectives and advantages of the present invention, some of which are specifically described and others that are not, will become apparent from the detailed description and claims that follow.

Detailed Description of the Invention : The following detailed description is provided to enable any person skilled in the art to which the present invention pertains to make and use the same, and sets forth the best mode contemplated by the inventors of carrying out the subject invention.

The present invention is the use of small organic molecules that have a pharmacological effect on cells and tissues to increase the enzymatic activity and/or expression of one or more membrane-type matrix metalloproteinases (MT-MMPs), or a similar enzyme, expressed in the trabecular meshwork (TM) of an eye to activate Gelatinase A (GelA) for the treatment of primary open angle glaucoma.

The use of the subject small organic molecules of the present invention for increasing cell membrane expression of MT-MMPs, and as a result, for activating GelA, increases the turnover and reduces the accumulation of extracellular matrix. Activating GelA in the TM increases outflow of aqueous and lowers intraocular pressure, thereby having therapeutic potential in the treatment of primary open angle glaucoma.

An additional use of the subject small organic molecules of the present invention is for increasing cell membrane expression of one or more MT- MMPs in the development of in vitro models, which could be used in the discovery of new medical treatments. In general, the small organic molecules of the present invention could contribute to the discovery of new treatments for any ocular disease with a pathophysiology involving changes in expression of one or more MT-MMPs and activation of GelA.

The small organic compounds of the present invention are generally represented by Figure 1 below : Figure 1 wherein W is selected from the group consisting of oxygen, sulfur and C 20 hydroxylated carbon group; Z is selected from the group consisting of oxygen and sulfur ; Ri is branched or unbranched and substituted or unsubstituted selected from the group consisting of C625 aryl such as for example but not limited to phenyl or naphthyl, C3-20 cycloalkyl such as for example but not limited to saturated bicyclic carbocyclic or tricyclic carbocyclic rings, C320 cycloalkenyl such as for example but not limited to unsaturated bicyclic carbocyclic or tricyclic carbocyclic rings, C3-30 heterocyclyl such as for example but not limited to 3- tetrahydrofuranyl, 2-tetrahydrothienyl, 4-piperidinyl, 2-pyrrolindinyl or 3- morpholinyl, C6-30 heteroaryl such as for example but not limited to furanyl, <BR> <BR> <BR> <BR> <BR> imidazolyl, quinolinyl, thiazolyi, indolyi, 3-thienyl, 2-benzofuranyl or 4-pyridyl, C6- 30 aryloxy such as for example but not limited to phenoxy, C630 arylalkoxy such as for example but not limited to phenylmethoxy, Ce-so aryloxycarbonyl such as for example but not limited to naphthyloxycarbonyl, C630 arylaminocarbonyl such as for example but not limited to naphthylaminocarbonyl, Ce-so arylalkyl such as for example but not limited to phenylmethyl, C630 aryloyl such as for example but not limited to benzoyl, Ce-so arylalkylcarbonyl such as for example but not limited to phenylmethylcarbonyl, C630 arylalkoxycarbonyl such as for example but not limited to phenylmethoxycarbonyl, C6-30 arylthioalkyl such as for example but not limited to phenylthiopropyl, C630 arylalkenyl such as for example but not limited to phenylpropenyl, C6-30 arylalkynyl such as for example but not limited to phenylethynyl, C6-30 aryloxyalkyl such as for example but not limited to phenoxymethyl, C6-30 arylalkylthioalkyl such as for example but not limited to phenylmeththiomethyl, C6-30 arylalkoxyalkyl such as for example but not limited to phenylpropoxyethyl, C630 arylthiocarbonyl such as for example but not limited to phenylthiocarbonyl, C630 arylalkylaminocarbonyl such as for example but not limited to naphthylpropylaminocarbonyl, C6-3o N-alkyl-N-arylaminocarbonyl such as for example but not limited to N-octyl-N-phenylaminocarbonyl, C6-30 N- arylaminosulfonyl such as for example but not limited to naphthylaminosulfonyl, C6-30 N-arylaminosulfinyl such as for example but not limited to N- naphthylaminosulfinyl, C6-30 arylsulfonyl such as for example but not limited to naphthylsulfonyl, C6-30 arylsulfinyl such as for example but not limited to naphthylsulfinyl, C6-30 aryloyloxy such as for example benzoyloxy, C6-30 arylthio such as for example phenylthio, C630 arylalkylthio such as for example but not limited to phenylmethylthio, C630 arylacyloxy, C630 arylalkylsulfonyl such as for example but not limited to phenylmethylsulfonyl, Ce-so arylalkylsulfinyl such as for example but not limited to naphthylpropylsulfinyl, C630 arylalkylthiocarbonyl such as for example but not limited to naphthylpropylthiocarbonyl, Ce-so N-alkyl-N- arylaminosulfonyl such as for example but not limited to N-octyl-N- phenylaminosulfonyl, C630 N-alkyl-N-arylaminosulfinyl such as for example but not limited to N-butyl-N-phenylaminosulfinyl, C6-3o aryloxycarbonylalkylcarbonyl such as for example but not limited to phenoxycarbonylmethylcarbonyl, C6-30 aryloylthio such as for example but not limited to naphthoyloylthio and Ce-so arylacylthio such as for example but not limited to benzoylthio, and Ri as defined above when substituted is substituted with one or more substituents selected from the group consisting of C1-25 alkyl such as for example but not limited to methyl, ethyl or propyl, C125 alkenyl such as for example but not limited to ethenyl, propenyl or octenyl, Ci-se alkynyl such as for example but not limited to ethynyl, butynyl or hexynyl, C125 alkoxy such as for example but not limited to methoxy, propoxy or butoxy, C125 alkenoxy such as for example but not limited to methenoxy, propenoxy or octenoxy, hydroxy, carboxy, amino, C1-25 (N- alkylcarbonyl) amino such as for example but not limited to (N- methylcarbonyl) amino, Ci-25 (N-alkylcarbonyl)-N-alkylamino such as for example but not limited to (N-methylcarbonyl)-N-propylamino, C125 (N- alkylcarbonylalkyl) amino such as for example but not limited to (N- methylcarbonylpropyl) amino, cyano, nitro, C6-30 arylazo such as for example but not limited to azobenzene, sulfo, sulfino, sulfhydryl, halo such as for example fluoro, chloro, bromo, or iodo, C125 haloalkyl such as for example but not limited to fluoromethyl or chloropropyl, trifluoromethyl, C1-25 trifluoromethylalkyl such as for example trifluoromethylpropyl, C6-3o arylalkyl such as for example but not limited to phenylmethyl, C6-30 aryl such as for example but not limited to phenyl or naphthyl, C125 N-alkylamino such as for example but not limited to N- methylamino, C125 N-dialkylamino such as for example but not limited to N- dimethylamino, C125 (N-alkyl-N-alkenyl) amino such as for example but not limited to (N-methyl-N-propenyl) amino, C1-25 N-dialkenylamino such as for example but not limited to N-dipropenylamino, C125 alkylsulfonyl such as for example but not limited to propylsulfonyl, C1-25 alkylsulfinyl such as for example but not limited to methylsulfinyl, C125 alkylthio such as for example but not limited to methylthio, hydrido, C1-25 cyanoalkyl such as for example but not limited to cyanobutyl, C125 acyl such as for example but not limited to acetyl, C1-25 alkylcarbonyloxy such as for example but not limited to methylcarbonyloxy, nitroso, C125 alkoxyalkyl such as for example but not limited to methoxypropyl, C125 alkoxycarbonyl such as for example but not limited to methoxycarbonyl, C 25 hydroxyalkyl such as for example but not limited to hydroxybutyl, thiocarboxy, C1-25 thiocarboxyalkyl such as for example but not limited to thiocarboxypropyl, C125 alkylthiocarbonyl such as for example but not limited to butylthiocarbonyl, Ci. alkylthiocarbonylalkyl such as for example but not limited to methylthiocarbonylethyl, C125 alkoxythiocarbonyl such as for example but not limited to methoxythiocarbonyl, C125 alkoxythiocarbonylalkyl such as for example but not limited to methoxythiocarbonylpropyl, sulfamoyl, sulfinamoyl, C125 N- alkylsulfamoyl such as for example but not limited to N-methylsulfamoyl, C125 N- dialkylsulfamoyl such as for example but not limited to N-dipropylsulfamoyl, C125 N-alkylsulfinamoyl such as for example but not limited to N-propylsulfinamoyl, C 25 N-dialkylsulfinamoyl such as for example but not limited to N- dimethylsulfinamoyl, C125 sulfamoylalkyl such as for example but not limited to sulfamoylmethyl, C125 sulfinamoylalkyl such as for example but not limited to sulfinamoylpropyl, aminocarbonyl, C125 aminocarbonylalkyl such as for example but not limited to aminocarbonylmethyl, C1-25 N-alkylaminocarbonyl such as for example but not limited to N-propylaminocarbonyl, C125 N-dialkylaminocarbonyl such as for example but not limited to N-dimethylaminocarbonyl, C125 alkoxycarbonylamino such as for example but not limited to propoxycarbonylamino, thiocarbamoyl, C125 thiocarbamoylalkyl such as for example but not limited to thiocarbamoylpropyl, C125 (N-alkyl) thiocarbamoyl such as for example but not limited to (N-methyl) thiocarbamoyl, C125 (N- dialkyl) thiocarbamoyl such as for example but not limited to (N- dimethyl) thiocarbamoyl, aminothio, C125 alkylaminothio such as for example but not limited to propylaminothio, C125 N-dialkylaminothio such as for example but not limited to N-dimethylaminothio, C125 cycloalkyl such as for example but not limited to saturated mono-, bi-or tricyclic carbocyclic rings, C125 cycloalkenyl such as for example but not limited to unsaturated mono-, bi-or tricyclic carbocyclic rings, C125 aryloxy such as for example but not limited to phenoxy, C125 cycloalkoxy such as for example but not limited to cyclopentoxy, C125 cycloalkenoxy such as for example but not limited to cyclopentenoxy, heterocycloxy, C125 heteroaryloxy, C125 heteroaryl such as for example but not limited to pyridyl, pyridino, morpholinyl or morpholino, heterocyclic amines, heterocyclic amides such as for example but not limited to piperidino or piperidinosulfamoyl, N-or S-oxo and N-or S-thioxo; R2 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C110 alkyl such as for example but not limited to methyl, propyl or butyl, C110 alkenyl such as for example but not limited to methenyl, propenyl or octenyl, C1-10 alkynyl such as for example ethynyl, butynyl or heptynyl, cyano, C110 haloalkyl such as for example but not limited to fluoromethyl, chloropropyl or fluorooctyl, trifluoromethyl, C615 cycloalkyl such as for example but not limited to cycloheptyl or cyclooctyl, C615 cycloalkenyl such as for example but not limited to cyclooctenyl or cyclononenyl, C615 heterocyclyl such as for example but not limited to 2-tetrahydrofuranyl, 3-tetrahydrothienyl, 4- piperidinyl, 2-pyrrolidinyl or 3-morpholinyl, C6-30 aryl such as for example but not limited to phenyl or naphthyl, C6-30 heteroaryl such as for example but not limited to furanyl, imidazolyl, quinolinyl, thiazolyl, indolyl, 2-thienyl, 3-benzofuranyl or 4- pyridyl, halo, nitro, Ce-so alkylaryl such as for example but not limited to methylphenyl or propylphenyl, C630 alkoxyaryl such as for example but not limited to ethoxyphenyl or butoxynaphthyl, C115 alkylcarbonyl such as for example but not limited to methylcarbonyl or propylcarbonyl, hydroxycarbonyl, aminocarbonyl, C115 N-alkylaminocarbonyl such as for example but not limited to N-butylaminocarbonyl or N-propylaminocarbonyl and C1-15 alkoxycarbonyl such as for example but not limited to methoxycarbonyl or propoxycarbonyl, and R2 as defined above may be substituted with one or more substituents as defined above for RI ; R3 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C112 alkyl such as for example but not limited to methyl or propyl, C112 alkenyl such as for example but not limited to ethenyl or butenyl, C112 alkynyl such as for example but not limited to octynyl or heptynyl, C120 alkoxyalkyl such as for example but not limited to methoxypropyl or ethoxybutyl, C120 alkoxyalkenyl such as for example but not limited to methoxypropenyl or ethoxybutenyl, C1-20 alkenoxyalkyl such as for example but not limited to methenoxypropyl or ethenoxybutyl and C120 alkenoxyalkenyl such as for example methenoxymethenyl or ethenoxypropenyl, and R3 as defined above may be substituted with one or more substituents as defined above for RI ; and R4 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, C112 alkyl such as for example but not limited to methyl or butyl, C1-12 alkenyl such as for example but not limited to ethenyl or propenyl, C1-12 alkynyl such as for example but not limited to methynyl or butynyl, C1-20 alkoxyalkyl such as for example but not limited to methoxypropyl, C1-20 alkoxyalkenyl such as for example but not limited to methoxypropenyl, C120 alkenoxyalkyl such as for example methenoxypropyl, Ci- 20 alkenoxyalkenyl such as for example methenoxypropenyl, Ce-so aryl such as for example but not limited to phenyl or naphthyl, Ce-so cycloalkyl such as for example but not limited to saturated mono-, bi-or tricyclic carbocyclic rings, Ce-so cycloalkenyl such as for example but not limited to unsaturated mono-, bi-or tricyclic carbocyclic rings, Ce-so heterocyclyl such as for example but not limited to 3-tetrahydrofuranyl, 2-tetrahydrothienyl, 3-piperidinyl, 2-pyrrolidinyl or 3- morpholinyl and Ce-so heteroaryl such as for example but not limited to furanyl, imidazolyl, quinolinyl, thiazolyl, indolyl, 3-thienyl, 2-benzofuranyl or 4-pyridyl, and R4 as defined above may be substituted with one or more substituents as defined above for Ri.

Most preferably, Ri as defined above is selected from the group consisting of phenyl, benzoyl, 2, 6-difluorophenyl, 4-trifluoromethylphenyl, 3,4- dimethoxyphenyl, 4-tert-butylphenyl, 4-methylphenyl, 4-methoxyphenyl, 4- phenoxyphenyl and 2-thienyl.

Most preferably, R2 as defined above is selected from the group consisting of cyano, cyclopentanyl, phenyl and trifluoromethyl.

Most preferably, R3 as defined above is selected from the group consisting of hydrido and alkyl.

Most preferably, R4 as defined above is selected from the group consisting of phenyl, cyclohexyl and alkyl.

Also preferable is R3 and R4 fused in a heterocyclic ring incorporating an amido nitrogen selected from the group consisting of pyrrolidino, piperidino, 4-methylpiperidino and morpholino.

In the case where R3 and R4 are fused in a heterocyclic ring, the same may also include substituents such as for example but not limited to hydrido, alkyl, alkenyl, alkynyl, alkoxyalkyl, alkoxyalkenyl, alkenoxyalkyl or alkenoxyalkenyl as described in detail above. An example not intended to be limiting would be the following glyoxylonitrile, aryl-, 0-carbamoyl oxime of Figure 2 below : Figure 2 Additional small organic compounds of the present invention are generally represented by Figure 3 below : Figure 3 wherein R6 is branched or unbranched and substituted or unsubstituted selected from the group consisting of C625 aryl such as for example but not limited to phenyl or naphthyl, C3-20 cycloalkyl such as for example but not limited to saturated bicyclic carbocyclic or tricyclic carbocyclic rings, C320 cycloalkenyl such as for example but not limited to unsaturated bicyclic carbocyclic or tricyclic carbocyclic rings, C330 heterocyclyl such as for example but not limited to 3- tetrahydrofuranyl, 2-tetrahydrothienyl, 4-piperidinyl, 2-pyrrolidinyl or 3- morpholinyl, C6-30 heteroaryl such as for example but not limited to furanyl, imidazolyl, quinolinyl, thiazolyl, indolyl, 3-thienyl, 2-benzofuranyl or 4-pyridyl, C6 30 arylalkyl such as for example but not limited to phenylmethyl, C630 arylthioalkyl such as for example but not limited to phenylthiopropyl, C6-30 arylalkenyl such as for example but not limited to phenylpropenyl, C6-30 arylalkynyl such as for example but not limited to phenylmethynyl, C630 aryloxyalkyl such as for example but not limited to phenoxymethyl, C6-30 arylalkylthioalkyl such as for example but not limited to phenylmeththiomethyl and C6-30 arylalkoxyalkyl such as for example but not limited to phenylpropoxyethyl, and R6 as defined above when substituted is substituted with one or more substituents selected from the group consisting of hydroxy, carboxy, amino, cyano, nitro, nitrate, C125 alkyl such as for example but not limited to methyl, ethyl or propyl, Ci-25 alkenyl such as for example but not limited to ethenyl, propenyl or octenyl, C125 alkynyl such as for example but not limited to ethynyl, butynyl or hexynyl, C125 alkoxy such as for example but not limited to methoxy, propoxy or butoxy, C125 alkenoxy such as for example but not limited to methenoxy, propenoxy or octenoxy, hydroxy, carboxy, amino, C125 (N- alkylcarbonyl) amino such as for example but not limited to (N- methylcarbonyl) amino, C125 (N-alkylcarbonyl)-N-alkylamino such as for example but not limited to (N-methylcarbonyl)-N-propylamino, C125 (N- alkylcarbonylalkyl) amino such as for example but not limited to (N- methylcarbonylpropyl) amino, C6-30 arylazo such as for example but not limited to azobenzene, sulfo, sulfino, sulfhydryl, halo such as for example but not limited to fluoro, chloro, bromo, or iodo, C1-25 haloalkyl such as for example but not limited to fluoromethyl or chloropropyl, trifluoromethyl, C1-25 trifluoromethylalkyl such as for example trifluoromethylpropyl, C630 arylalkyl such as for example but not limited to phenylmethyl, C6-30 aryl such as for example but not limited to phenyl or naphthyl, C125 N-alkylamino such as for example but not limited to N- methylamino, C125 N-dialkylamino such as for example but not imited to N-dimethylamino, C125 (N-alkyl-N-alkenyl) amino such as for example but not limited to (N-methyl-N-propenyl) amino, C1-25 N-dialkenylamino such as for example but not limited to N-dipropenylamino, 61-25 alkylsulfonyl such as for example but not limited to propylsulfonyl, C125 alkylsulfinyl such as for example but not limited to methylsulfinyl, C125 alkylthio such as for example but not limited to methylthio, hydrido, C125 cyanoalkyl such as for example but not limited to cyanobutyl, C125 acyl such as for example but not limited to acetyl, C1-25 alkylcarbonyloxy such as for example but not limited to methylcarbonyloxy, nitroso, C125 alkoxyalkyl such as for example but not limited to methoxypropyl, C125 alkoxycarbonyl such as for example but not limited to methoxycarbonyl, C 25 hydroxyalkyl such as for example but not limited to hydroxybutyl, thiocarboxy, C1-25 thiocarboxyalkyl such as for example but not limited to thiocarboxypropyl, C125 alkylthiocarbonyl such as for example but not limited to butylthiocarbonyl, C125 alkylthiocarbonylalkyl such as for example but not limited to methylthiocarbonylethyl, C125 alkoxythiocarbonyl such as for example but not limited to methoxythiocarbonyl, C1-25 alkoxythiocarbonylalkyl such as for example but not limited to methoxythiocarbonylpropyl, sulfamoyl, sulfinamoyl, C125 N- alkylsulfamoyl such as for example but not limited to N-methylsulfamoyl, C125 N- dialkylsulfamoyl such as for example but not limited to N-dipropylsulfamoyl, C125 N-alkylsulfinamoyl such as for example but not limited to N-propylsulfinamoyl, C 25 N-dialkylsulfinamoyl such as for example but not limited to N- dimethylsulfinamoyl, C1-25 sulfamoylalkyl such as for example but not limited to sulfamoylmethyl, C125 sulfinamoylalkyl such as for example but not limited to sulfinamoylpropyl, aminocarbonyl, C125 aminocarbonylalkyl such as for example but not limited to aminocarbonylmethyl, C125 N-alkylaminocarbonyl such as for example but not limited to N-propylaminocarbonyl, C125 N-dialkylaminocarbonyl such as for example but not limited to N-dimethylaminocarbonyl, C125 alkoxycarbonylamino such as for example but not limited to propoxycarbonylamino, thiocarbamoyl, C125 thiocarbamoylalkyl such as for example but not limited to thiocarbamoylpropyl, C125 (N-alkyl) thiocarbamoyl such as for example but not limited to (N-methyl) thiocarbamoyl, C125 (N- dialkyl) thiocarbamoyl such as for example but not limited to (N- dimethyl) thiocarbamoyl, aminothio, C125 alkylaminothio such as for example but not limited to propylaminothio, C125 N-dialkylaminothio such as for example but not limited to N-dimethylaminothio, C125 cycloalkyl such as for example but not limited to saturated mono-, bi-or tricyclic carbocyclic rings, C1-25 cycloalkenyl such as for example but not limited to unsaturated mono-, bi-or tricyclic carbocyclic rings, C125 aryloxy such as for example but not limited to phenoxy, C125 cycloalkoxy such as for example but not limited to cyclopentoxy, C125 cycloalkenoxy such as for example but not limited to cyclopentenoxy, heterocycloxy, C125 heteroaryloxy, C125 heteroaryl such as for example but not limited to pyridyl, pyridino, morpholinyl or morpholino, heterocyclic amines, heterocyclic amides such as for example but not limited to piperidino or piperidinosulfamoyl, N-or S-oxo and N-or S-thioxo; Y is selected from the group consisting of O and S; the R7 groups may be the same or different selected from the group consisting of H or C1-15 alkyl ; R8 is branched or unbranched and substituted or unsubstituted selected from the group consisting of hydrido, trifluoromethyl, cyano, C1-12 alkyl such as for example but not limited to methyl or butyl, C112 alkenyl such as for example but not limited to methenyl or propenyl, C1-12 alkynyl such as for example but not limited to methynyl or butynyl, C125 alkoxy such as for example but not limited to methoxy, propoxy or butoxy, C125 alkenoxy such as for example but not limited to methenoxy, propenoxy or octenoxy, C125 alkynoxy such as for example but not limited to methynoxy, propynoxy or octynoxy, C125 alkylthio such as for example but not limited to methylthio, propylthio or octylthio, C125 alkylcarboxy such as for example but not limited to methylcarboxy, propylcarboxy or octylcarboxy, C1-25 alkenylcarboxy such as for example but not limited to methenylcarboxy, propenylcarboxy or octenylcarboxy, C1-25 alkynylcarboxy such as for example but not limited to methynylcarboxy, propynylcarboxy or octynylcarboxy, C125 aminoalkyl such as for example but not limited to aminomethyl, aminopropyl or aminooctyl, C125 alkylaminoalkyl such as for example but not limited to propylaminomethyl, ethylaminopropyl or butylaminooctyl, C1-25 dialkylaminoalkyl such as for example but not limited to dipropylaminomethyl, diethylaminopropyl or dibutylaminooctyl, C630 aryl such as for example but not limited to phenyl or naphthyl, Ce-so cycloalkyl such as for example but not limited to saturated mono-, bi-or tricyclic carbocyclic rings, Ce-so cycloalkenyl such as for example but not limited to unsaturated mono-, bi-or tricyclic carbocyclic rings, C630 heterocyclyl such as for example but not limited to 3-tetrahydrofuranyl, 2-tetrahydrothienyl, 3- piperidinyl, 2-pyrrolidinyl or 3-morpholinyl, C6-30 heteroaryl such as for example but not limited to furanyl, imidazolyl, quinolinyl, thiazolyl, indolyl, 3-thienyl, 2- benzofuranyl or 4-pyridyl, Ce-so arylalkyl such as for example but not limited to phenylmethyl or naphthylpropyl, C630 cycloalkylalkyl such as for example but not limited to saturated mono-, bi-or tricyclic alkylated carbocyclic rings, Ce-so cycloalkenylalkyl such as for example but not limited to unsaturated mono-, bi-or tricyclic alkylated carbocyclic rings, C6-30 cycloalkylcarboxy such as for example but not limited to cyclopentylcarboxy, C6-30 cycloalkenylcarboxy such as for example but not limited to cyclopentenylcarboxy, C6-30 heterocyclyalkyl such as for example but not limited to heterocyclyoctyl, C630 heteroarylalkyl such as for example but not limited to heterophenylmethyl, C6-30 arylcarboxy such as for example but not limited to naphthylcarboxy, C6-30 heterocyclylcarboxy, and C630 heteroarylcarboxy such as for example but not limited to heteronaphthylcarboxy, and R8 as defined above may be substituted with one or more substituents as defined above for R6 ; R9 is either Rio is selected from the group consisting of O, S, CH-NO2, N-N02, N-S (0) 2-CH3 and N-CN; Q is NR$R$ whereby the Rs groups may be fused in a heterocyclic ring, the same may also include substituents such as for example but not limited to hydrido, Cul-12 alkyl, C112 alkenyl, C112 alkynyl, C125 alkoxyalkyl, C125 alkoxyalkenyl, Cl-25 alkenoxyalkyl or Cl-25 alkenoxyalkenyl as described in detail above; m is an integer less than 3; and n is an integer less than 7.

Preferred molecules having the general structure of Figure 3 above include for example but are not limited to molecules where R6 is naphthyl (alpha- or beta-) or 2, 3-dichlorophenyl, Y is oxygen, both R7 groups are hydrido, n is two or three, R8 is isopropyl and Rg is where Rio is oxygen and Q is NR8R8 with one R8 group hydrido and the other hydrido or methyl. Additionally, certain molecules of the present invention may exist in different isomeric forms. The present invention contemplates all such isomers both individually in pure form, and in admixture, including racemic mixtures.

Additional small organic compounds of the present invention are generally represented by Figure 4 below : Figure 4 wherein R6 is the same as defined for R6 of Figure 3 and may be substituted with one or more substituents as defined above for R6 ; Y is the same as defined for Y of Figure 3 above; RI, is selected from the group consisting of hydrido, C1-15 branched or straight alkyl and Ci-ie branched or straight alkenyl ; R12 is selected from the group consisting of hydrido and C1-15 alkyl ; L is selected from the group consisting of nitrate, YR13 and NR8R9 ; R13 is selected from the group consisting of C115 alkyl, C115 alkenyl, C115 alkynyl, C115 haloalkyl, trifluoromethyl and cyanomethyl ; R8 is the same as defined for Rs of Figure 3 and may be substituted with one or more substituents as defined above for R8 ; Rg, Rio, Q and m are the same as defined for Rg, Rio, Q and m respectively of Figure 3 above; q is an integer less than 5; and t is an integer less than 9.

Additional compounds of the present invention based on principles of nonclassical bioisosteres, are illustrated by the general structures of Figures 5 through 8 below : Figure 5 Figure 6 Figure 7 Figure 8 wherein W, Z, Ri, R2, R3, and R4 are as defined above in Figure 1; U is sulfinyl or sulfonyl ; and R5 is selected from the group consisting of hydrido, cyano, halo, trifluoromethyl, haloalkyl, alkyl, alkenyl, alkynyl, aryl, aryloxy, alkoxy, alkenoxy, cycloalkyl, cycloalkenyl, cycloalkoxy, cycloalkenoxy, heterocyclyl, heterocycloxy, heteroaryl and heteroaryloxy, as further defined for substituents of Ri in Figure 1 above.

Certain small organic compounds of the present invention may exist in different isomeric forms, in particular, isomers related to an oxime nitrogen-carbon bond, which are depicted in Figures 1 through 8 by the symbol The invention contemplates all such isomers both individually, in pure form and in admixture, including racemic mixtures.

Examples of small organic molecules of the present invention include but are not limited to adenylyl cyclase (AC) inhibitors such as for example but not limited to 9- (tetrahydro-2'-furyl) adenine, 2', 5'-dideoxyadenosine and miconazole, phospholipase D (PLD) activators such as but not limited to roxithromycin, fluoride ion, bradykinin, progesterone, endothelin, vasopressin, 4- hydroxynonenal, interleukin-11, angiotensin 11, N- (6-aminohexyl)-5-chloro-1- naphthalenesulfonamide and oxidized low density lipoprotein, cyclic adenosine monophosphate (cAMP) phosphodiesterase (CAP) activators such as for example but not limited to phosphatidic acids such as dioleoyl, dioctanoyl and 1- stearoyl-2-arachidonyl-sn-glycerol-3-phosphate, phosphatidic acid analogues such as thiophosphatidic acid and phosphatidic acid (PA) containing alkyl ether, or vinyl ether linkages rather than ester bonds and pyrazinyl guanidine, protein kinase A inhibitors such as for example but not limited to (N- [2- ( (p- bromocinnamyl) amino) ethyl]-5-isoquinolinesulfonamide, protein phosphatase inhibitors such as for example but not limited to vanadium salts such as potassium bisperoxo (1, 10-phenanthroline) oxovanadate and dipotassium bisperoxo (picolinate) oxovanadate, molybdat oxoanions, tungstate oxoanions and dephostatins such as 3, 4-dihydroxy-N-methyl-N-nitrosoaniline and 3,6- dihydroxy-N-methyl-N-nitrosoaniline, phosphatidate phosphohydrolase inhibitors/cationic amphiphiles such as for example but not limited to propranolol, tetracaine, mepacrine, desmethylimipramine, chlopromazine and desipramine, Rho activators such as for example but not limited to sphingosine-1-phosphate, lipid lowering agents or hyperlipoproteinemics such as for example but not limited to 2-tetradecylglycidic acid, 5- (tetradecyloxy)-2-furoic acid, 3-thiadicarboxylic acid, 3- (4-methylpiperazin-1-yl)-1-phenylpropanone, 6,7-dihydro-5H- dibenz [c, e] azepine, N-2-n-butylindazolone, 4-phenyl-5, 5-dicarbethoxy-2- pyrrolidinone, 4- (4-hydroxy-3-iodophenoxy) 3,5-diiodohydrocinnamic acid, 1- methyl-4-piperidyl bis (p-chlorophenoxy) acetate, 2- [ [l-methyl-2- [3- (trifluoromethyl) phenyl] ethyl] amino] ethanol benzoate ester and 5- methylpyrazinecarboxylic acid 4-oxide and concanavalin A (Con A) receptor ligands such as for example but not limited to acetylcholinesterase and complement protein 1q.

The small organic molecules of the present invention are described in still greater detail in the examples that follow.

EXAMPLE 1: Small molecules that increase expression or activity of a cell- associated component with gelatinase activity, detected using a cell-based screening assay that measures hydrolysis of a thiopeptolide substrate.

A. Thiopeptolide Assay used to detect cell-associated gelatinase activity ascribed to MT-MMP: Rhesus monkey TM cells are cultured and maintained for at least two weeks in 96-well microtiter plates in a growth medium such as Dulbecco's Modified Eagles'Medium (DMEM) plus 15 percent (v/v) fetal bovine serum (FBS) containing 1 % bovine calf serum or a medium more suitable for endothelial cells due to its lower serum content such as MCDB 131 supplemented with endothelial cell growth supplement, 1 % or less FBS or bovine calf serum and defined supplements as described by Knedler and Ham, In Vitro Cellular and Dev. Biol. 23: 481 (1987). Two days before molecule testing, the medium is replace by a defined, serum- free medium such as Minimum Essential Medium (MEM) containing defined supplements as described by Schachtschabel and Binninger, Z. f. Gerontol. 26: 243 (1993), but preferably using a basal medium such as MCDB 131 containing defined supplements, because of the absence of interfering substances that could bind or compete with test compounds and because of its ability to maintain endothelial-like cells such as TM cells as a stable, nonproliferative monolayer while optimizing expression of native structural and functional attributes. Unless specifically noted, all molecules tested were prepared as stock solutions in dimethyl sulfoxide (DMSO) with a final concentration of 10 mg/ml. The stocks were stored in a dessicator at-20 degrees Celsius. In testing the molecules, the molecules were diluted to final concentrations of 0.3 ug/ml and 15 ug/ml in a simplified culture medium based on Ames'medium, called Concanavalin A (Con A) conditioning medium (CACM). The monkey TM cells were incubated with the test molecules for 48 hours. Control medium of CACM plus DMSO and a positive control with 5 ug/ml Con A plus DMSO were run in parallel. At the end of the incubation period, the experimental media were replaced with 100 uL of the buffer that is part of the thiopeptolide assay mixture (50 mmol/L HEPES, 5 mmol/L Catch, 3.5 mmol/L KCI, 106 mmol/L NaCI, 0.02% (v/v) Brij 35, pH 7.5). Next 100uL of a doubly (2X) concentrated mixture of the thiopeptolide substrate (from a forty times (40X) concentrated DMSO stock, to give 1 mmol/L final concentration) freshly combined with the thiol reagent (5,5'dithiobis (2- nitrobenzoic acid (DTNB) from a twenty times (20X) concentrated DMSO stock to give1 mmol/L) were added to each well and were incubated at 37 degrees Celsius for two hours with gentle agitation.

At the end of the incubation period, by means of a spectrophoto-metric plate reader, the optical density (OD) at 410 nm was determined for each well after automatic subtraction of a blank value for a well containing reaction mixture but without cells. The average OD at the two-hour end point for each test molecule at all concentrations in triplicate was calculated. This calculation was interpreted to be a measure of cell- associated MT-MMP level, equivalently defined as either its activity (a catalytic property of the enzyme) or expression (number of functional molecules). The percent difference in OD for each sample compared to the CACM control reflects the effectiveness of each test molecule or combination of molecules in eliciting increases in MT-MMP levels. As an alternate to the end point OD reading, one may use the mean rate of appearance of the reaction product (mean V), calculated from the best linear fit of the data to absorbance vs. time. In so doing, measurements are taken every five minutes with the first and possibly last time points, that do not contribute to a good linear fit, routinely eliminated.

B. Testing of small organic molecules for associated MT-MMP activity: TM cells were exposed to molecules and combinations of molecules by means of simultaneous addition, from separate stocks, to incubation medium.

1. Piperidyl carbonyl nitrile oxime molecules : Piperidyl carbonyl nitrile oxime molecules are synthesized as described in U. S. Patent Number 4,453, 974 incorporated herein in its entirety by reference. Results from molecules tested are set forth below in Table 1.

Table 1 Molecule Conc. umol/L Percent Activation Over control value 1 0. 9 48. 1 Me 47. 3. 1 260. 4 Me0 0 O CN 2 1. 0 33. 8 51. 1. 1 420. 5 F 0 0 O, F CN F Cf 3 0. 9 14. 8 42. 9. 1 350. 1 'o PYRO \o N'V CN 2. Aryloxydialkylurea molecules : Aryloxydialkylurea molecules are synthesized as described in U. S. Patent Number 5,021, 429 incorporated herein in its entirety by reference. Results from molecules tested are set forth below in Table 2.

Table 2 Molecule Conc. umol/L Percent Activation Over control value 4 1. 0 5. 9 49. 6 73. 4 OH i-Pr i pN\/I3H2 \. I O O Table 2-Continued Molecule Conc. umol/L Percent Activation Over control value 5 1. 1 (-16. 4) OH 57. 0 94. 9 Wo\o, o~ y/\i n li 6 1. 0 2. 2 52. 4 207. 9 i-Pr 9wO wNNH z ° \ 7 1. 0 (-7. 9) 49. 2 274. 0 . .-Pr c 1 fizz C, NU Cl 3 3 3. Guanabenz molecules : Guanabenz molecules are commercially available from Sigma Aldrich Corporation, St. Louis, Missouri. Results from molecules tested are set forth below in Table 3.

Table 3 Molecule Conc. umol/L Percent Activation Over control value 8 2. 6 2. 9 ci 12. 98 12. 7 NH N 64. 91 54. 3 CH NH NH Ci 129. 81 106. 8 9 1. 18 21. 4 118. 44 82. 4 NH XT L 2 Table 3-Continued Molecule Conc. umol/L Percent Activation Over control value 10 0. 3 (-0. 5) cl 76. 28 75. 5 NH \ I . CH NH NH2 2 11 1. 14 (-20. 9) 57. 22 88. 1 Nu NU _N CH NHs CH CI 12 1. 05 3. 2 52. 53 101. 6 ci NH \ I NH \ NH'NH NH 2 2 C1 OH 4. Tricyclic molecules : Tricyclic molecules are commercially available from Sigma Aldrich Corporation, St. Louis, Missouri. Results from molecules tested are set forth below in Table 4.

Table 4 Molecule Conc. umol/L Percent Activation Over control value 13 1. 1 74. 6 55. 9 68. 5 I C. N S 14 1. 27 7. 7 63. 49 115. 6 3 C ONH2 Regarding negative OD readings in screening assay for MT-MMP, since some color development takes place in the controls, the control OD readings are above zero reflecting a constitutive level of MT-MMP expression.

Negative values may likewise result from an artifactual interference in reading the OD at 410 nm, due in part to absorbance properties of the test compound, or from a loss of cells in the well. It is likewise possible that as a result of incubation with a test compound at a particular concentration for 48 hours, there was a net loss of MT-MMP protein molecules or reduced activity of this MMP compared with the control.

As illustrated by the results set forth above, certain structural characteristics are apparently crucial for activity: 1) the piperidine ring; 2) the carbonyl-oxime backbone; 3) the nitrile function; and 4) an aromatic ring attached to the same oxime carbon as the nitrile. Likewise, certain functional groups attached to the aromatic ring increase the activity of the compound.

EXAMPLE 2: Small molecules that have a synergistic effect when used in combination, with each other or with the lectin Concanavalin A, on the increased expression or activity of a cell-associated component with gelatinase activity, detected using a cell-based screening assay that measures hydrolysis of a thiopeptolide substrate.

Concanavalin A (Con A) binds to mannose groups on cell membrane gycoproteins. Because of its tetravalent nature, Con A binding leads to oligomerization of these"receptors,"possibly in the form of heterogeneous aggregates. Con A stimulation elicits increased expression of MT-MMP with concomitant ability to activate proGel A. The response, as measured by the magnitude of zymographic bands and by thiopeptolide assay, is dose-dependent.

Thus the effects of co-activation using a submaximal dose of Con A plus treatment with a small molecule that may or may not be active by itself, are measured in the subject activation assay.

A. Co-activation assay used to detect cell-associated gelatinase activity ascribed to MT-MMP: For the coactivation assays, following the preliminary steps set forth above preceding incubation with test compounds in Example 1, cells subsequently undergo one of the following protocols.

Protocol 1. Cells are incubated with a particular test compound for twenty-four hours. This incubation period is followed by a second twenty-four hour incubation period, in which the culture medium containing the first test compound is completely replaced by medium containing 5ug/ml Con A.

Protocol 2. Cells are incubated with a particular first test compound for twenty-four hours, followed by a second forty-eight hour incubation in which the medium containing the first test compound is completely replaced by medium containing either Con A, as described above, or a second test compound.

Protocol 3. Cells are incubated with a combination of a test compound together with 5 ug/ml Con A for forty-eight hours.

Two controls, with or without 5 ug/ml Con A are run in parallel as described in Example 1. After incubation with these agents, cells are processed in equivalently to those in Example 1. Instead of comparison of the OD values with a CACM control, the results are compared with samples using Con A alone, or with a single test compound as described in Protocol 2 above, as set forth in Table 5 below.

Table 5 Molecule Protocol Conc. umol/L Activity (% increase (with Con A unless over Con A above, or as other- noted otherwise) wise noted) 6 1 104.8 183.9 6 2 34.9 49.0 6 2 100.0 70.9 7 2 49.1 59.8 6 and 3 2 34.9 : 14.3 18.1 (above 3 only) 6 and 3 2 100.0 : 42.9 39.6 (above 3 only) 13 1 1.1 18.1 13 1 55.9 109.5 A convenient form for administering one or more small organic molecules of the present invention to increase Gelatinase A activity in ocular cells is through a pharmaceutically acceptable composition comprising one or more of the following : one or more small organic molecules or one or more hydrates of the molecules ; one or more small organic molecules or one or more acid addition salts of the molecules whereby suitable acids include for example but are not limited to mineral acids such as hydrohalic acids, organic acids such as acetic acid, or acids which are sparingly soluble and impart slow-release properties to their salts, such as pamoic acid; and one or more small organic molecules or one or more base addition salts of the molecules whereby suitable salts include those formed from inorganic bases such as hydroxides, carbonates, bicarbonates, or alkoxide of the alkali or alkaline earth metals, organic bases such as mono-, di-, and trialkylamines, alkanolamines, alkene-diamines, phenylalkylamines, cyclic saturated bases, cyclic unsaturated bases or alkylamines forming quaternary salts.

The organic bases forming such salts are of suitable molecular size to be therapeutically acceptable. Acid and base addition salts in accordance with the present invention are prepared by conventional means known by those skilled in the art.

If such pharmaceutical acceptable compositions are formulated as a sterile solution or a suspension in water or other aqueous media, the above formulation would likewise include physiological salt solutions whereby the pH is suitably adjusted and/or buffered and the tonicity is suitably adjusted for optimal absorption, distribution, release, and/or efficacy at the site of action on or within the eye.

If such pharmaceutically acceptable compositions are formulated as a non-aqueous solution or suspension, the above formulation would likewise include an oil, an organic solvent or methyl sulfoxide. Also, formulations of the present invention could likewise include cyclodextrin, a detergent or other non- toxic pharmaceutical excipients combined covalently or noncovalently with a biodegradable or a nonerodable encapsulating substance such as a polymer, as known to those skilled in the art.

The small organic compounds of the present invention are administered to treat glaucoma through a method of delivery to the tissues of the trabecular meshwork of the eye. Methods of such delivery of one or more of the small organic molecules of the present invention include for example but are not limited to application of externally applied eye drops, ointments or implants, injection or insertion a solution, suspension, or sustained-release implant into the anterior chamber or sclera of an eye, external application on the scleral surface of an eye and/or administration as an adjunct pharmaceutical treatment at the time of surgical treatment for glaucoma, as with filtration surgery. Various forms of delivery include single or multiple dosages such that an acute, short term therapy schedule performed once or intermittently over a specified time frame could be useful as an alternative to sustained therapy. In this way, the compounds could be useful for effecting varying degrees of amplification of aqueous outflow through the trabecular meshwork and adjoining structures.

While there is described herein certain specific embodiments of the present invention, it will be manifest to those skilled in the art that various modifications may be made without departing from the spirit and scopeof the underlying inventive concept and that the same is not limited to the particular forms herein described except insofar as indicated by the scope of the appended claims.