USE OF PHOSPHODIESTERASE INHIBITORS AND CYCLIC NUCLEOTIDE ANALOGUES FOR THE MANUFACTURE OF A MEDICAMENT FOR STIMULATING TEAR SECRETION

Abstract: For the stimulation of tear secretion, a preparation is top­ ically applied to the ocular surface, the preparation including a phosphodiesterase inhibitor and/or a cyclic nucleotide ana­ logue. Inhibition of phosphodiesterase and cyclic nucleotide analogues increases cyclic nucleotide levels in accessory lac­ rimal gland tissue, thereby stimulating tear secretion. The preparation additionally contains a vehicle for the phospho­ diesterase inhibitor or cyclic nucleotide analogue, and may contain an ophthalmic preservative.

Description: This invention relates to medicaments for stimulating tear secretion. More particularly, it relates to the stimulation of tear secretion with topically applied agents including certain phosphodiesterase inhibitors and certain cyclic nucleotide analogues that increase intracellular cyclic nucleotide levels in lacrimal gland tissue. There are a number of situations where it is desirable to increase the amount and/or to modify the nature of tear fluid produced by the eye. Illustrative instances include the treatment of a spectrum of dry eye disorders including, but not limited to, keratocon junctivitis sicca, agerelated dry eye, Stevens-Johnson syndrome, ocular cicatrical pemphigoid blepharitis, neurotrophic ocular surface disease and corneal exposure. In addition, patients who wear contact lenses may have sub-optimal rates of tear production for optimal contact lens wear. Increased tear production is likely to increase eye comfort and contact lens comfort, and improve contact lens wear. These patients will therefore benefit from agents that increase tear production. The lacrimal gland is an exocrine gland which secretes protein and, by different mechanisms, water and electrolytes. To stimulate secretion, agonists are employed which increase intracellular free calcium concentration, and/or the concentration of adenosine 3′,5′-cyclic monophosphate (cyclic AMP or cAMP). However, proteins are secreted by exocytosis, whereas electrolytes and water are secreted as the permeability of cell membranes is selectively increased to sodium, potassium and chloride. A given agonist could stimulate water and electrolyte secretion, but not protein secretion, and visa versa. The cyclic nucleotides, cyclic AMP and guanosine 3′,5′-cyclic monophosphate (cyclic GMP or cGMP) are known to regulate a variety of metabolic processes. Cyclic AMP has been suggested to function as a second messenger for exocytosis in the lacrimal gland. Phosphodiesterases are enzymes responsible for the destruction of cyclic AMP by catalyzing the hydrolytic reaction of cyclic AMP and water to produce adenosine 5′-monophosphate and for the destruction of cyclic GMP by catalyzing the hydrolytic reaction of cyclic GMP and water to produce guanosine 5′-monophosphate. Various molecular forms of phosphodiesterase are distributed in tissues and cells. Agents that inhibit phosphodiesterases may produce tissue selective responses (Weishaur, R.E. et al J. of Med. Chem. 28; 537-545, 1985). It is known that the phosphodiesterase inhibitors theophylline, 3-isobutyl-1-methylxanthine (IBMX), and pentoxyfylline increase cyclic AMP levels in lacrimal gland cells and stimulate protein secretion from lacrimal gland tissue pieces or cells in vitro (Friedman ZY et al, Biochemica et Biophysica Acta 675: 40-45, 1981: Jahn R. et al, Eur. J. Biochem 126: 623-629, 1982; Mauduit P. et al, Am. J. Physiol J. 246; C37-C44, 1984; Dartt DA et al, J. Physiol (London) 352; 375-384, 1984). Cellular cyclic nucleotide levels can also be increased by exogenous addition of cyclic nucleotide analogues to which cells are permeable. Cells are not permeable to cyclic AMP or cyclic GMP, thus the exogenous addition of these compounds does not elevate cellular cyclic nucleotide levels. It is known that exogenous 8-bromo cyclic AMP, dibutyryl cyclic AMP or adenosine 3′-5′-cyclic phosphorothioate stimulate protein secretion from lacrimal gland tissue pieces or cells in vitro, but 8-bromo cyclic GMP does not. (Friedman ZY et al, Biochemica et Biophysica Acta 675: 40-45, 1981: Jahn R. et al, Eur. J. Biochem 126: 623-629, 1982; Mauduit P. et al, Am. J. Physiol J. 246; C37-C44, 1984; Dartt DA et al, J. Physiol (London) 352; 375- 384, 1984). What is needed are agents which will, in vivo, stimulate tear secretion upon topical administration to the ocular surface. A topical mode of administration has several advantages. It eliminates the need for injections in patients with dry eye disorders, thereby decreasing systemic effects, cost of therapy, and the amount of drug needed. Accordingly, we have sought to provide an improved method for stimulating tear secretion by topical administration. In particular, we have sought to provide an improved method of stimulating lacrimal gland secretion by topical application of compounds to the ocular surface. We have aimed to provide for the treatment of dry eye disorders, more particularly to facilitate eye treatment without systemic therapy such as injection. An object of this invention is to provide an improved agent for topical application to improve eye comfort, e.g. to enhance contact lens wear and comfort. Another object is to provide a preparation for increasing the amount of the tear fluid produced by accessory lacrimal gland tissue. In accordance with the invention, it has been found that agents including phosphodiesterase inhibitors and cyclic nucleotid analogues that increase intracellular cyclic nucleotide levels in lacrimal gland tissue effectively stimulate tear secretion when applied topically to the eye. This method for stimulating tear secretion is useful in the treatment of dry eye disorders, and it facilitates such treatment by eliminating the need for injections. Also, a corresponding preparation has been developed which can be applied topically to stimulate lacrimal gland secretion. A preferred preparation contains a phosphodiesterase inhibitor or a cyclic nucleotide analogue and a preservative. The preparation typically also contains a physiologically compatible vehicle. FR-A-2,358,155 relates to a cyclic 3′, 5′ adenosine monophosphate derivative, specifically a butyryl derivative, which is alleged to cure inflammation upon topical administration. This cAMP derivative can optionally be admixed with a phosphodiesterase inhibiting amount of aminophylline. The reference claims that the composition acts by reducing the histidine level upon topical administration. This effect is not related to, and the reference nowhere contemplates, the stimulation of tear secretion from accessory lacrimal glands. The present invention comprises use of a compound selected from phosphodiesterase inhibitors and cyclic nucleotide analogues, for the manufacture of a topical preparation for administration to the ocular surface to stimulate in vivo fluid secretion from the human accessory lacrimal glands. The compound may for example include (a) phosphodiesterase inhibitors such as 3-isobutyl-1-methylxanthine (IBMX), and theophylline, and well as caffeine and theobromine and other methylxanthine phosphodiesterase inhibitors that increase cyclic nucleotide levels in lacrimal gland tissue, and (b) cyclic nucleotide analogues such as 8-bromo cAMP, dibutyryl cAMP, adenosine 3′,5′-cyclic phosphorothioate, 8-bromo cGMP, and other cyclic nucleotide derivatives. These compounds may be used alone or in combination with one another. The compounds deemed preferred are IBMX and 8-bromo cAMP. A medicament made by use of the present invention can comprise an amoun of the said compound, which consists of a phosphodiesterase inhibitor, which is effective to increase intracellular cyclic nucleotide levels in the accessory lacrimal gland of the end so as to promote fluid secretion from the said gland without effecting the main lacrimal gland; an ophthalmic preservative; and a physiologically compatible vehicle selected from polyethers, polyvinyls, cellulose derivatives, mineral oil, white petroleum, polymers of acrylic acid, lanolin and polysaccharides. Application of a medicament made by use of the invention involves topical administration to the ocular surface of compounds that increase intracellular cyclic nucleotid levels, thereby stimulating tear secretion. Several modes of topical administration may be used in the practice of the invention. For example, the compounds may be administered topically to the eye as drops, or within ointments, gels, or liposomes. Further, the compounds may be infused into the tear film by means of a pump-catheter system. In other embodiments, the compounds are attached to and/or incorporated into or carried by contact lenses or contained within continuous or other selective-release devices including membranes, for example microcapsule membranes, for contact with the ocular surface. The development of the foregoing agents that are effective in stimulating lacrimal secretion when applied topically to the eye is unexpected for several reasons. First, the main lacrimal gland is not exposed to the surface of the eye and lies separated from the ocular surface by a relatively great diffusion distance. The main lacrimal gland is connected to the surface only through a series of microscopic ducts. Therefore, while drugs injected vascularly can reach the main lacrimal gland parenchyma, it is considered unlikely that topically applied drugs will do so. Second, although there are microscopic nests of accessory lacrimal gland tissue within the conjunctiva, one would expect that insufficient lacrimal tissue mass will preclude the success of an approach that seeks to act merely on the accessory glands. The practice of this invention will now be explained in more detail by way of example only in the following description, and with reference to the accompanying drawings, in which: Figures 1A, 1B and 1C represent results obtained from practice of the invention with rabbits, the Figures being graphs of the osmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing IBMX in concentrations of respectively 0.3 mM, 1.0 mM and 3.0 mM; Figures 2A, 2B, 2C and 2D likewise represent results obtained from practice of the invention with rabbits, the Figures being graphs of the osmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing theophylline in concentrations of 3.0 mM, 10.0 mM, 30.0 mM, and 60.0 mM; Figures 3A, 3B and 3C represent results obtained from further practice of the invention with rabbits: Figure 3A being a graph of the osmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing 3X10 _6 M glucagon; Figure 3B being a graph of the oxmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing 0.5 mM of IBMX; and Figure 3C being a graph of the osmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing 3X10 _6 M glucagon and 0.1 mM IBMX; Figures 4A, 4B and 4C represent results obtained from further practice of the invention with rabbits, the Figures being graphs of the osmolality of tear samples as a function of time taken from a rabbit eye treated with a buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing 8-bromo cAMP in concentration of 1.0 mM, 3.0 nM, and 10.0 nM; and Figures 5A, 5B and 5C represent results obtained from further practice of the invention with rabbits; the Figures being graphs of the osmolality of tear samples as a function of time taken from a rabbit eye treated with buffer solution and, subsequently, the same eye treated in accordance with the invention with buffer solution containing 8-bromo cGMP in concentrations of 1.0 mM, 3.0 mM, and 10.0 mM. In accordance with the invention, tear secretion is stimulated with topically applied agents including phosphodiesterase inhibitors, and cyclic nucleotid analogues that increase intra-cellular cyclic nucleotide levels in lacrimal gland tissue. More particularly, these phosphodiesterase inhibitors can include methylxanthines including IBMX and theophylline. By way of specific example, the cyclic nucleotide analogues can include 8-bromo cyclic AMP, and 8-bromo cyclic GMP. A further practice employs a combination of a phosphodiesterase inhibitor with a gastrointestinal hormone and/or biologically active peptide that activates vasoactive intestinal peptide or malantropin receptor. A preparation according to the invention can, by way of illustration, be applied to the eye in animals and humans as a drop or within ointments, gels, or liposomes. Further, the compounds may be infused into the tear film via a pump-catheter system. In other embodiments, the compounds can be contained within continuous or other selective-release devices, for example, membranes, such as those used in the Ocusert (Trade Mark) system (Alza Corp., Palo Alto, CA). The "Ocusert" system has a pilocarpin/alginic acid mixture in a vinyl acetate copolymer membrane, and is but one example of a device which releases a drug at a controlled, hopefully zero order kinetic rate. As a further specific example, the compounds can be attached to or carried by and/or contained within contact lenses that are placed on the eye. In general, it is desired that the mode of application be such that the compounds enter the tear film or make contact with the surface of the eye. In vivo examples in accordance with the invention were conducted on rabbits with dry eyes. The dry eye disorder was created by surgically closing the duct that carries fluid from the main lacrimal gland to the tear film, and surgically removing the nictitans and harderian glands. This leaves intact only the accessory glands that lie on the ocular surface. These rabbits develop increased tear film osmolality soon after the operation, a finding that is understood to be due to decreased tear production, and that is characteristic of dry eye. It is recognized that results of ophthalmologic tests using rabbits has close correlation with humans and therefore the results carry over to humans. The effect of topically applied isotonic buffer solution with and without methylxanthine phosphodiesterase inhibitors or cyclic nucleotide analogues on tear film osmolality was studied in the dry eye rabbit. All test drops were ten microlitres (µl) in volume. Tear samples were taken with a micropipette system, in the manner described in the article entitled "Osmolarity of Tear Micro-volumes in Keratoconjunctivities Sicca", by Gilbard et al, Arch Ophthalmol 96: 677, 1978. Osmolality was measured by freezing-point depression. The following protocol was used to test the effects of topically applied methylxanthines or cyclic nucleotide analogues. At zero time a tear sample was taken for measurement of osmolality. At one minute the test drop was instilled. At six and at sixteen minutes, tear samples were taken for osmolality measurements. The above sequence was repeated several times, each time after a wait of five minutes after the sixteen-minute sample, to test additional drops. The following drops were instilled in the test eye and after each such instillation test samples were taken as stated: 1) buffer solution, 2) buffer solution containing a methylxanthine or cyclic nucleotide analogue at a low dose, 3) buffer solution, 4) buffer solution containing a methylxanthine or cyclic nucleotide analogue at a moderate dose, 5) buffer solution, and 6) buffer solution containing a methylxanthine or cyclic nucleotide analogue at a higher dose. In addition, one example (Figure 2) also included the following: 7) buffer solution, and 8) buffer solution containing a methylxanthine at a still higher dose. The results are shown in Figures 1A, 1B, and 1C, Figures 2A, 2B, 2C and 2D, Figures 3A, 3B, and 3C, Figures 4A, 4B and 4C, and Figures 5A, 5B and 5C; where Figures 1, 2, 3, 4 and 5 each represent results obtained from the same rabbit. Figure 1 represents results obtained with IBMX. The concentrations of IBMX given in the 10 µl drops were: A) 0.3 mM, B) 1.0 mM, and C) 3.0 mM. As shown in Figure 1, buffer solution containing IBMX lowers the tear film osmolality more effectively than buffer alone. The Figure also indicates a dose-dependent decrease in tear film osmolality, and the decrease was strikingly more pronounced than the effect of the buffer alone. This reflects an IBMX stimulated dose-dependent increase in tear secretion. Figure 2 represents results obtained with theophylline. The concentrations of theophylline given in the 10 µl drops were: A) 3.0 mM, B) 10.0 mM, C) 30.0 mM, and D) 60.0 mM. As shown in Figure 2, buffer solution containing 60.0 mM theophylline lowers tear film osmolality more effectively than buffer alone. This reflects a theophylline stimulated increase in tear secretion. Figure 3 represents results obtained with IBMX, glucagon, and a combination of IBMX and glucagon Glucagon is a gastrointestinal peptide which upon topical administration to the ocular surface produces a dose-dependent decrease in tear film osmolality. IBMX and glucagon were tested in combination to determine if the two have a synergisticeffect on tear film osmolality. The concentrations given in 20 µl drops were as follows: A) 3X10 _6 M glucagon, B) 0.5 mM of IBMX, 3) 3X10 _6 M glucagon and 0.5 mM IBMX. As shown in Figure 3, the combination of glucagon and IBMX lowers tear film osmolality more effectively than either glucagon or IBMX alone. Figure 4 represents results obtained with 8-bromo cAMP. The concentrations of 8-bromo cAMP given in the ten microliter drops were (a) 1.0 mM, (b) 3.0 mM, and (c) 10.0 mM. As shown in Figure 4, buffer solution containing 8-bromo cAMP lowers tear film osmolality more effectively than buffer alone. This reflects an 8-bromo cAMP stimulated increase in tear secretion. Figure 5 represents results obtained with 8-bromo cGMP. The concentrations of 8-bromo cGMP given in the ten microliter drops were (a) 1.0 mM, (b) 3.0 mM, and (c) 10.0 mM. As shown in Figure 5, buffer solution containing 8-bromo cGMP lowers tear film osmolality more effectively than buffer alone. This reflects an increase in tear secretion stimulated by the 8-bromo cGMP. Further in accordance with the invention, a topical tear-stimulation preparation was made by combining a phosphodiesterase inhibitor or cyclic nucleotide analogue that increases cyclic nucleotide levels in lacrimal gland tissue with an appropriate preservative. The preparation typically also contains a physiologically compatible vehicle, as those skilled in the art can select using conventional criteria. The vehicles may be selected from the known ophthalmic vehicles which include water, polyethers such as polyethylene glycol, polyvinyls such as polyvinyl alcohol and povidone, cellulose derivatives such as methylcellulose, and hydroxypropyl methylcellulose, petroleum derivatives such as mineral oil, white petrolatum, animal fats such as lanolin, vegetable fats such as peanut oil, polymers of acrylic acid such as carboxypolymethylene gel, polysaccharides such as dextrans, glycosaminoglycans such as sodium hyaluronate and salts such as sodium chloride and potassium chloride. One preferred vehicle is the non-toxic ophthalmic preparation which has the following composition: 22.0 to 43.0 millimoles of potassium per liter; 29.0 to 50.0 millimoles of bicarbonate per liter; 130.0 to 140.0 millimoles of sodium per liter; and 118.0 to 136.5 millimoles of chloride per liter Phosphodiesterase inhibitor preservatives include sorbic acid, chlorohexidine, benzalkonium chloride and thimerosal. Preservatives for cyclic nucleotide analogue include chlorohexidine. Other preservatives may be appropriate, as those skilled in the art can select using conventional criteria. Stated generally, both the vehicle and preservative are to be physiologically compatible with the phosphodiesterase inhibitors and cyclic nucleotide analogues and are not to inactivate the secretion-stimulating activity of these compounds. In accordance with the invention, one example of the preparation contains IBMX, the foregoing non-toxic ophthalmic preparation as a vehicle, and chlorohexidine for installation in the eye in drop form. Another example consists 8-bromo cAMP, the foregoing non-toxic ophthalmic preparation and chlorohexidine. It will thus be seen that the objects set forther above, among those made apparent from the preceding description, are efficiently attained.

Claims: 1. Use of a compound selected from phosphodiesterase inhibitors and cyclic nucleotide analogues, for the manufacture of a topical preparation for administration to the ocular surface to stimulate in vivo fluid secretion from the human accessory lacrimal glands. 2. The use according to claim 1, wherein the phosphodiesterase inhibitors are selected from methylxanthine, caffeine, theophylline, and derivatives thereof. 3. The use according to claim 1, wherein the cyclic nucleotide analogues are selected from bromine derivatives of cyclic AMP, butyryl derivatives of cyclic AMP, bromine derivatives of cyclic GMP, and adenosine 3′-5′ cyclic phosphorothioate. 4. The use according to claim 3, wherein the cyclic nucleotide analogues are selected from 8-bromo cyclic AMP, 8-bromo cyclic GMP, and dibutyryl cyclic AMP. 5. The use according to any of claims 1 to 4, wherein the selected compound is used in admixture with glucagon for the manufacture of the said preparation. 6. The use according to any of claims 1 to 5, wherein the topical preparation is included in a pump-catheter system, a selected release device, or a contact lens. 7. The use of a compound according to claim 1 for the manufacture of a topical preparation which consists of a phosphodiesterase inhibitor and is dispersed in a carrier vehicle selected from drops of liquid, gels, ointments, and liposomes. 8. The use according to claim 7, comprising an amount of the said compound which is effective to increase intracellular cyclic nucleotide levels in the accessory lacrimal gland of the eye so as to promote fluid secretion from the said gland without effecting the main lacrimal gland; an ophthalmic preservative; and a physiologically compatible vehicle selected from polyethers, polyvinyls, cellulose derivatives, mineral oil, white petroleum, polymers of acrylic acid, lanolin, and polysaccharides. 9. The use according to claim 7 or claim 8, wherein the phosphodiesterase inhibitor is selected from methylxanthine, caffeine, theophylline, and derivatives thereof. 10. The use according to claim 7, 8 or 9, which further includes glucagon.

Inventors: DARTT, DARLENE A., GILBARD, JEFFREY P.
Applicants: EYE RESEARCH INSTITUTE OF  RETINA FOUNDATION
Application year: 1987
Main classification: A61K31
IPC codes: A61K45, A61P27, A61K31, A61P43