Treatment of glaucoma using phosphodiesterase inhibitor.

0001At the Association for Research in Vision and Ophthalmology Meeting May 1-6, 1988, Poster No. 22 was entitled "Ocular Hypotension Induced by Grisfolic Acid, a Phosphodiesterase Inhibitor" by Hiromu Mishima et al..

Background of the Invention

0002In the sympathetic nervous system cascade, adenosine triphosphate (ATP) is converted by action of a catecholamine, such as epinephrine, to cyclic adenosine monophosphate (cAMP) which is then converted to 5′-adenosine monophosphate (5′-AMP) by the action of the enzyme phosphodiesterase (PDE). This cascade results in physiological responses to environmental stresses, the so-called adrenergic effects.

0003PDE inhibitors include papaverine and the xanthines such as theophylline and caffeine. PDE inhibitors are used in cosmetics as set forth in U.S. Patent 3,978,213. Theophylline as well as other PDE inhibitors are used as a bronchodilator for relief of the symptoms of asthma.

0004Glaucoma is an ocular disorder most often characterized by increased intraocular pressure which, over time, may cause impaired vision or blindness. Treatments include topical agents such as pilocarpine (a cholinomimetic drug); timolol maleate (a β-adrenergic receptor blocking agent); epinephrine (an α- and β-adrenergic receptor agonist); dipivefrin (a prodrug of epinephrine) and demecarium bromide (a cholineesterase inhibitor). Systemic agents used to treat glaucoma include carbonic anhydrase inhibitors such as acetazolamide.

0005An object of the present invention is an effective treatment for glaucoma by providing an agent which lowers intraocular pressure.

Summary of the Invention

0006A treatment for lowering intraocular pressure is provided whereby a phosphodiesterase inhibitor is administered to a patient.

Detailed Description of the Invention

0007Particular PDE inhibitors include compounds of the following formula (I): <chemistry id="chem0001" num="0001"><img file="EP0345028A2_D0001.tif" /></chemistry> wherein X may be H and H or O; R¹ may be H, C₁₋₆alkyl, C₃₋₆branched-chain alkyl or C₃₋₆cycloalkyl; R² may be H, C₁₋₆alkyl, C₃₋₆branched-chain alkyl, C₃₋₆cycloalkyl or C₂₋₆alkenyl; R³ may be H, C₁₋₆alkyl, C₃₋₆branched-chain alkyl or C₃₋₆cycloalkyl, and when X is 2H, R³ may also be acyl, arylacyl or alkanesulfonyl; R⁴ may be H, halogen, C₁₋₆alkyl, C₃₋₆branched-chain alkyl, C₃₋₆cycloalkyl or C₁₋₆alkoxy; R⁵ and R⁶ may each be H, C₁₋₆alkyl, C₃₋₆branched-chain alkyl or C₃₋₆cycloalkyl; and the dotted line may be a single or double bond between C4 and C5 of the pyridazine ring.

0008Preferred compounds of formula (I) are those wherein R¹ is CH₃, R² and R³ are hydrogen, R⁴, R⁵ and R⁶ are hydrogen or CH₃, X is O and the pyridazinone ring is attached at C-7 of the benzoxazine ring. A particular compound of formula (I) is 6-[3,4-dihydro-3-oxo-1,4[2H]-­benzoxazine-7-yl]-2,3,4,5-tetrahydro-5-methylpyridazin-­3-one known as ORF 22,867 and having the following formula (Ia): <chemistry id="chem0002" num="0002"><img file="EP0345028A2_D0002.tif" /></chemistry>

0009The above compounds of formula (I) are described in U.S. Patent No. 4,721,784 to Donald W. Combs, issued January 26, 1988 which is hereby incorporated by reference with respect to synthesis and definition of the compounds.

0010A second particular group of PDE inhibitors to be used in the present invention are quinazolinones of the following formula (II): <chemistry id="chem0003" num="0003"><img file="EP0345028A2_D0003.tif" /></chemistry> wherein R⁷ is hydrogen provided that R⁷ is hydrogen only when the dotted line linkage represents a saturated bond; R⁸ is hydrogen, lower alkyl, cycloalkyl having 4-8 carbon atoms, cycloalkylalkyl wherein the cycloalkyl group has 4-8 carbon atoms and the alkyl group has 1-3 carbon atoms, haloalkyl having 1-3 halogen atoms and 1-4 carbon atoms, norbornyl and norbornylmethyl; R⁹ is lower alkyl, and the dotted line linkage represents a saturated bond or a double bond.

0011A preferred compound of formula (II) is 5,6-dimethoxy-­4-methyl-2[1H]-quinazolinone known as ORF 16,600 or bemarinone and having the following formula (IIa): <chemistry id="chem0004" num="0004"><img file="EP0345028A2_D0004.tif" /></chemistry>

0012The above compounds of formula (II) are described in U.S. Patent No. 4,490,374 to Victor T. Bandurco et al. issued December 25, 1984 which is hereby incorporated by reference with respect to synthesis and definition of the compounds.

0013Other specific PDE inhibitors to be used in the present invention include those described by el Allaf in Arch. Int. Physiol. Biochim., Nov. 1984, 92(4) S 69-79 and by W. S. Hillis in Eur. Heart, J., Dec. 1982, 3 Suppl D, 97-101, including carbazeran. Systemic formulations of PDE inhibitors are prepared as known in the art, e.g. as set forth in U.S. Patent 4,721,784.

0014For topical application, the PDE inhibitor may be formulated by preparing a solution or suspension which may be preserved with a preservative such as benzalkonium chloride and formulated with aids such as polyvinyl alcohol or methyl cellulose. The PDE inhibition may be used in an amount of 0.05 to 2.0% by weight of the ophthalmic composition. The suspension formulation may be similar to that of Dexacidin made by Iolab Pharmaceuticals of Claremont, California with 6 mg/mL hydropropylmethylcellulose, 8.5 mg/mL sodium chloride, 0.5 mg/mL of Polysorbate 20 and 0.04 mg/mL benzalkonium chloride with water q.s. and HCl to adjust the pH to 5.3 to 5.7. Solubility may be enhanced by formulation with a nonionic or anionic surfactant, particularly a nonionic surfactant, such as Pluronic F-68 or by complexation with a cyclodextrin.

0015Drug delivery systems for use in the present invention include eye drops, ointments and controlled release systems such as Ocusert system from Alza Corporation of Palo Alto, California and those described in my co-pending application, U.S. Serial No. 61,591, filed June 15, 1987, which is equivalent to EPO Publication No. 251,680, published February 24, 1988.

0016The dosage to be used will depend on the severity of the patient's condition, the potency of the particular PDE inhibitor and the formulation used. For example, a potent PDE inhibitor may be used at a high dose with, however, a controlled release system as described above. PDE inhibitory activity may be determined in vitro in accordance with the method of W. J. Thompson, et al., in <u style="single">Adv. Cycli. Nucleotide Res.</u> Ed. G. Brooker, et al., Vol. 10, pp. 69-92 (1979). This assay measures the ability of compounds to inhibit cyclic nucleotide phosphodiesterase. This enzyme converts either cyclic AMP or cyclic GMP to the noncyclized AMP or GMP, respectively. Compounds are tested at various concentrations in the presence of cyclic AMP (0.10-1.0 µM containing 0.2 µCi ³H-cyclic AMP), enzyme, and 0.05M Tris-Cl buffer (pH 7.4, containing 5mM MgCl₂). After a specified time, the reaction is stopped by heating to 100^oC for one minute. After cooling, 0.10 ml of a solution containing snake venom (1 mg/ml) is added and the reaction is allowed to proceed for 30 minutes. Termination of this reaction is accomplished by the addition of 1.0 ml of 33% Dowex slurry to separate the product from unconverted substrate. An aliquot is removed from the supernatant and quantitated by liquid scintillation spectrometry. The results are expressed as the IC₅₀ which is the concentration (µM) of compound required to inhibit 50% of the cyclic nucleotide phosphodiesterase activity.

0017In vivo lowering of intraocular pressure may be tested by the method of Goldmann Aplanation Tonometry, Schiotz Tonometry or other systems such as the Tono-Pen or the pneumotonometer.

EXAMPLE 1

0018To test PDE inhibitors as agents for lowering intraocular pressure, normotensive and hypertensive rabbits were used. Active agents were formulated by preparing a sterile suspension of the agent at a 2% by weight concentration in aqueous saline. Each preparation was coded and tested in conjunction with a placebo which is randomly applied to the normotensive eye. In the hypertensive rabbit, only one eye will be used. Intraocular pressure is determined by either the Tono-Pen hand-held tonometer made by Intermedics Intraocular of Pasadena, California or a pneumotonometer with topical anesthetic. Data is analyzed with the paired T-test.

0019Using the above procedure, ORF 16,600 test results were as shown below in Table I: <tables id="tabl0001" num="0001"><table frame="all"><title>Table I</title><tgroup cols="5" colsep="1" rowsep="0"><colspec colnum="1" colname="col1" colwidth="31.50mm" /><colspec colnum="2" colname="col2" colwidth="31.50mm" /><colspec colnum="3" colname="col3" colwidth="31.50mm" /><colspec colnum="4" colname="col4" colwidth="31.50mm" /><colspec colnum="5" colname="col5" colwidth="31.50mm" /><thead valign="top"><row><entry namest="col1" nameend="col1" align="center"><u style="single">Hr</u></entry><entry namest="col2" nameend="col2" align="center"><u style="single">Drug</u></entry><entry namest="col3" nameend="col3" align="center"><u style="single">Placebo</u></entry><entry namest="col4" nameend="col4" align="center"><u style="single">Delta</u></entry><entry namest="col5" nameend="col5" align="left"><u style="single">p</u></entry></row></thead><tbody valign="top"><row><entry namest="col1" nameend="col1" align="right">0</entry><entry namest="col2" nameend="col2" align="right">20.3 ± 2.5</entry><entry namest="col3" nameend="col3" align="right">20.0 ± 2.9</entry><entry namest="col4" nameend="col4" align="char" char=".">+0.3</entry><entry namest="col5" nameend="col5" align="char" char=".">.39</entry></row><row><entry namest="col1" nameend="col1" align="right">1</entry><entry namest="col2" nameend="col2" align="right">17.0 ± 2.8</entry><entry namest="col3" nameend="col3" align="right">17.8 ± 2.3</entry><entry namest="col4" nameend="col4" align="char" char=".">-0.8</entry><entry namest="col5" nameend="col5" align="char" char=".">.08</entry></row><row><entry namest="col1" nameend="col1" align="right">2</entry><entry namest="col2" nameend="col2" align="right">17.1 ± 3.3</entry><entry namest="col3" nameend="col3" align="right">17.2 ± 2.0</entry><entry namest="col4" nameend="col4" align="char" char=".">-0.1</entry><entry namest="col5" nameend="col5" align="char" char=".">.86</entry></row><row><entry namest="col1" nameend="col1" align="right">3</entry><entry namest="col2" nameend="col2" align="right">19.7 ± 2.7</entry><entry namest="col3" nameend="col3" align="right">20.0 ± 2.6</entry><entry namest="col4" nameend="col4" align="char" char=".">-0.3</entry><entry namest="col5" nameend="col5" align="char" char=".">.47</entry></row><row rowsep="1"><entry namest="col1" nameend="col1" align="right">4</entry><entry namest="col2" nameend="col2" align="right">20.3 ± 3.1</entry><entry namest="col3" nameend="col3" align="right">20.5 ± 1.8</entry><entry namest="col4" nameend="col4" align="char" char=".">-0.2</entry><entry namest="col5" nameend="col5" align="char" char=".">.79</entry></row></tbody></tgroup></table></tables>

0020The relatively weak potency of ORF 16,600 reflected in Table I appears to be caused by a lack of solubility may be enhanced by increasing the solubility of the drug, e.g. by complexing with a β-cyclodextrin or a poloxomer.