Methods of Preparing Ophthalmic Formulations and Uses of Same

Abstract

The present invention relates to a process for preparing an ointment comprising subjecting a mixture of a fluorescent molecular rotor compound, mineral oil, and grinding media to ball milling agitation and combining this mixture with a hydrophobic vehicle for an ointment. The invention also relates to an ophthalmic formulation comprising a pharmaceutically acceptable carrier and a compound of the following structural Formula (I), or a pharmaceutically acceptable salt thereof: Values and preferred values of the variables in structural Formula (I) are described herein.

What is claimed is: 
     
         1 . A process for preparing an ophthalmic ointment, comprising:
 (a) combining a fluorescent molecular rotor compound with mineral oil as a levigating agent, thereby obtaining a first mixture;   (b) subjecting the first mixture to ball milling agitation with a grinding media, thereby obtaining a second mixture; and   (c) combining the second mixture with a hydrophobic vehicle, thereby obtaining an ointment.   
     
     
         2 . The process of  claim 1 , wherein the second mixture comprises particles of the fluorescent molecular rotor compound, the particles having a size distribution characterized by a d(0.95) particle size of less than about 20 microns. 
     
     
         3 . The process of  claim 1 , wherein the grinding media for ball milling agitation comprises beads with an average size of at least 3 mm. 
     
     
         4 . The process of  claim 1 , wherein the ratio of the grinding media volume to the ball milling container volume ranges from about 0.05 to about 0.35. 
     
     
         5 . The process of  claim 1 , wherein the grinding media is a ceramic selected from the group consisting of magnesia stabilized zirconia, zirconium silicate, zirconia stabilized alumina, alumina-zirconia composite, burundum, and yttrium stabilized zirconia. 
     
     
         6 . The process of  claim 5 , wherein the grinding media is yttrium stabilized zirconia. 
     
     
         7 . The process of  claim 1 , wherein the second mixture is agitated at about 50% to about 70% of critical mill speed. 
     
     
         8 . The process of  claim 1 , wherein the hydrophobic vehicle is selected from one of more of: a petrolatum, a mineral oil, a polyglycol, a silicone oil, a fluorocarbon, a lanolin, a gelled mineral oil, and a surfactant. 
     
     
         9 . The process of  claim 8 , wherein the hydrophobic vehicle is selected from one or more of a petrolatum and a mineral oil. 
     
     
         10 . The process of  claim 1 , further comprising irradiating the fluorescent molecular rotor compound and the mineral oil before forming the first mixture. 
     
     
         11 . The process of  claim 1 , further comprising irradiating the first mixture. 
     
     
         12 . The process of  claim 1 , wherein the fluorescent molecular rotor compound has the following structural Formula (I), or a pharmaceutically acceptable salt thereof: 
       
         
           
           
               
               
           
         
         wherein:
 A 1  is an optionally substituted C6-C 18 arylene, an optionally substituted C5-C 18 heteroarylene, or is represented by the following structural formula: 
 
       
       
         
           
           
               
               
           
         
         R 1  and R 2  are each independently hydrogen, optionally substituted C1-C12 alkyl, an optionally substituted C1-C12 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or R 1  and R 2  taken together with the nitrogen atom to which they are attached form an optionally substituted 3 to 12 membered heterocycloalkyl; 
         R 3  and R 4  are each independently hydrogen, methyl, or ethyl; 
         R 5  is —OH, optionally substituted —O(C1-C6 alkyl), —NR 6 R 7  or is represented by the following structural formula: 
       
       
         
           
           
               
               
           
         
         R 6  and Ware each independently, hydrogen, methyl, ethyl or R 6  and R 7  taken together with the nitrogen atom to which they are attached form a 5 to 7 membered heterocycloalkyl containing one to three ring heteroatoms independently selected from N, O, and S; 
         wherein:
 y is an integer from 1 to 10; 
 R 8 , for each occurrence independently, is hydrogen, —OH, or —CH 2 OH; 
 R 9  is hydrogen, —NR 10 R 11 , —C(O)R 12 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl; 
 R 10 , R 11 , and R 12  are each independently hydrogen or C1-C6 alkyl. 
 
       
     
     
         13 . The process of  claim 12 , wherein A 1  is selected from the group consisting of optionally substituted phenyl, optionally substituted naphthyl, an optionally substituted (E)-stilbene, or an optionally substituted (Z)-stilbene. 
     
     
         14 . The process of  claim 12 , wherein A 1  is optionally substituted naphthyl. 
     
     
         15 . The process of  claim 12 , wherein R 1  and R 2  taken together with the nitrogen atom to which they are attached form an optionally substituted 3 to 12 membered heterocycloalkyl. 
     
     
         16 . The process of  claim 12 , wherein R 5  is 
       
         
           
           
               
               
           
         
       
     
     
         17 . The process of  claim 12 , wherein R 5  is 
       
         
           
           
               
               
           
         
         y is 3; and 
         R 9  is methyl. 
       
     
     
         18 . The process of  claim 12 , wherein the fluorescent molecular rotor compound has the following structural Formula (II) or Formula (III), or a pharmaceutically acceptable salt thereof: 
       
         
           
           
               
               
           
         
         wherein:
 R 13 , R 14  and R 15  are each independently hydrogen, —OH, or optionally substituted —O(C1-C6 alkyl). 
 
       
     
     
         19 . The process of  claim 1 , wherein the fluorescent molecular rotor compound is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         20 . A process for preparing an ophthalmic ointment, comprising:
 (a) combining a fluorescent molecular rotor compound of the following structural formula:   
       
         
           
           
               
               
           
         
         
           with mineral oil as a levigating agent, thereby obtaining a first mixture; 
         
         (b) subjecting the first mixture to ball milling agitation, thereby obtaining a second mixture; and 
         (c) combining the second mixture with a hydrophobic vehicle, thereby obtaining an ointment. 
       
     
     
         21 . An ophthalmic ointment comprising an effective amount of a compound of the following structural Formula (I), or a pharmaceutically acceptable salt thereof: 
       
         
           
           
               
               
           
         
         wherein:
 A 1  is an optionally substituted C6-C 18 arylene, an optionally substituted C5-C18 heteroarylene, or is represented by the following structural formula: 
 
       
       
         
           
           
               
               
           
         
         
           R 1  and R 2  are each independently hydrogen, optionally substituted C1-C12 alkyl, an optionally substituted C1-C12 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or R 1  and R 2  taken together with the nitrogen atom to which they are attached form an optionally substituted 3 to 12 membered heterocycloalkyl; 
           R 3  and R 4  are each independently hydrogen, methyl, or ethyl; 
           R 5  is —OH, optionally substituted —O(C1-C6 alkyl), —NR 6 R 7  or is represented by the following structural formula: 
         
       
       
         
           
           
               
               
           
         
         
           R 6  and Ware each independently, hydrogen, methyl, ethyl or R 6  and R 7  taken together with the nitrogen atom to which they are attached form a 5 to 7 membered heterocycloalkyl containing one to three ring heteroatoms independently selected from N, O, and S; 
           wherein:
 y is an integer from 1 to 10; 
 R 8 , for each occurrence independently, is hydrogen, —OH, or —CH 2 OH; 
 R 9  is hydrogen, —NR 10 R 11 , —C(O)R 12 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl; 
 R 10 R), R 11  and R12 are each independently hydrogen or C1-C6 alkyl; 
 
         
         and a pharmaceutically acceptable carrier. 
       
     
     
         22 . An ophthalmic ointment comprising an effective amount of a compound of the following structural Formula (II): 
       
         
           
           
               
               
           
         
         wherein:
 R 1  and R 2  are each independently hydrogen, optionally substituted C1-C12 alkyl, an optionally substituted C1-C12 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or R 1  and R 2  taken together with the nitrogen atom to which they are attached form an optionally substituted 3 to 12 membered heterocycloalkyl; 
 R 3  and R 4  are each independently hydrogen, methyl, or ethyl; 
 R 5  is —OH, optionally substituted —O(C1-C6 alkyl), —NR 6 R 7 , or is represented by the following structural formula: 
 
       
       
         
           
           
               
               
           
         
         
           R 6  and R 7  are each independently, hydrogen, methyl, ethyl, or R 6  and R 7  taken together with the nitrogen atom to which they are attached form a 5 to 7 membered heterocycloalkyl containing one to three ring heteroatoms independently selected from N, O, and S; 
           R 10 , R 11  and R 12  are each independently hydrogen or C1-C6 alkyl; 
           R 13  is hydrogen, —OH, or optionally substituted —O(C1-C6 alkyl); 
           wherein:
 y is an integer from 1 to 10; 
 R 8 , for each occurrence independently, is hydrogen, —OH, or —CH 2 OH; 
 R 9  is hydrogen, —NR 10 R 11 , —C(O)R 12 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl; 
 
         
         and a pharmaceutically acceptable carrier. 
       
     
     
         23 . An ophthalmic ointment obtained by the process according to  claim 1 . 
     
     
         24 . A method of diagnosing a neurodegenerative disease in a subject, comprising:
 administering to the subject an effective amount of the ophthalmic ointment of  claim 21 .   
     
     
         25 . The method of  claim 24 , wherein the neurodegenerative disorder is an amyloidogenic disorder. 
     
     
         26 . The method of  claim 25 , wherein the amyloidogenic disorder is selected from Alzheimer's Disease (AD), Familial AD, Sporadic AD, Creutzfeld-Jakob disease, variant Creutzfeld-Jakob disease, spongiform encephalopathies, Prion diseases (including scrapie, bovine spongiform encephalopathy, and other veterinary prionopathies), Parkinson's disease, Huntington's disease (and trinucleotide repeat diseases), amyotrophic lateral sclerosis, Down's Syndrome (Trisomy 21), Pick's Disease (Frontotemporal Dementia), Lewy Body Disease, neurodegeneration with brain iron accumulation (Hallervorden-Spatz Disease), synucleinopathies (including Parkinson's disease, multiple system atrophy, dementia with Lewy Bodies, and others), neuronal intranuclear inclusion disease, tauopathies (including progressive supranuclear palsy, corticobasal degeneration, hereditary frontotemporal dementia (with or without Parkinsonism), and Guam amyotrophic lateral sclerosis/parkinsonism dementia complex. 
     
     
         27 . A method of diagnosing a neurodegenerative disease in a subject, comprising:
 administering to the subject an effective amount of the ophthalmic ointment of  claim 22 .   
     
     
         28 . The method of  claim 27 , wherein the neurodegenerative disorder is an amyloidogenic disorder. 
     
     
         29 . The method of  claim 28 , wherein the amyloidogenic disorder is selected from Alzheimer's Disease (AD), Familial AD, Sporadic AD, Creutzfeld-Jakob disease, variant Creutzfeld-Jakob disease, spongiform encephalopathies, Prion diseases (including scrapie, bovine spongiform encephalopathy, and other veterinary prionopathies), Parkinson's disease, Huntington's disease (and trinucleotide repeat diseases), amyotrophic lateral sclerosis, Down's Syndrome (Trisomy 21), Pick's Disease (Frontotemporal Dementia), Lewy Body Disease, neurodegeneration with brain iron accumulation (Hallervorden-Spatz Disease), synucleinopathies (including Parkinson's disease, multiple system atrophy, dementia with Lewy Bodies, and others), neuronal intranuclear inclusion disease, tauopathies (including progressive supranuclear palsy, corticobasal degeneration, hereditary frontotemporal dementia (with or without Parkinsonism), and Guam amyotrophic lateral sclerosis/parkinsonism dementia complex. 
     
     
         30 . The method of  claim 23 , comprising:
 determining the amount of amyloid aggregate in the supranuclear region and/or cortical lens region of the eye.