Cannabinoid for use in treating eye disorders

Abstract

Disclosed herein are methods for treating eye disorders. The methods include administering to a subject in need of treatment a therapeutically effective amount of Compound (I) having the following formula or a pharmaceutically acceptable salt thereof.

Description

CANNABINOID FOR USE IN TREATING EYE DISORDERSRELATED APPLICATIONS

[0001] This patent application claims the benefit of priority to U.S serial no.63 / 537,491, filed on, September 9, 2023.FIELD OF THE INVENTION

[0002] The present invention is in the field of treating eye disorders using a peripherally restricted cannabinoid.BACKGROUND OF THE DISCLOSURE

[0003] There is no known cure for glaucoma. However, lowering intraocular pressure (IOP) is helpful to slow or arrest the progression of the disease. Reduction of IOP can be achieved with pharmaceutical, laser, or surgical interventions. Drugs that are commonly used to treat glaucoma include prostaglandin F2a analogues, Rho kinase inhibitors, P-blockers, a2-agonists, carbonic anhydrase inhibitors, and cholinergic agonists. A meta-analysis determined that over 50% of glaucoma patients require two or more drugs to successfully reduce IOP (Tanna et al., 2010). This mercurial efficacy also coincides with several side-effects associated with each class of drugs. Aside from medications, surgeries may also reduce IOP through enhancement of aqueous humor outflow. Despite this, not all surgeries are successful, and some may only modestly reduce IOP, with some patients requiring additional treatment after surgery. Glaucoma surgeries may also increase the risk of intraocular infections (Gedde et al., 2001). One of the biggest challenges in treating glaucoma is poor patient compliance to topical glaucoma medication (Newman-Casey et al., 2015). People, especially in the age range when glaucoma is diagnosed and treated, often experience difficulty administering eye drops, so finding an oral form of treatment may increase patient compliance.

[0004] Cannabinoids have been shown to consistently reduce IOP. When given orally, A-9-THC reduced IOP 30 minutes to 3 hours after administration (Flach, 2002; Hepler and Petrus, 1976; Hepler et al., 1976; Tomida et al., 2006). The synthetic A-l-THC cannabinoid BW146Y dose-dependently reduced IOP after 1-hour post-administration (Tiedeman et al., 1981). Dronabinol and nabilone (synthetic cannabinoid) reduced IOP after 2 hours (Newell et al., 1979; Plange et al., 2007). Oral dosing of palmitoylethanolamide reduced IOP in clinical studies (Costagliola et al., 2014; Gagliano et al., 2011; Pescosolido et al., 2011; Rossi et al., 2020; Strobbe et al., 2013). When inhaled, cannabis products or cigarettes containing A-9-THC reduced IOP within 30 80 minutes (Flom et al., 1975; Hepler and Frank, 1971; Hepler and Petrus, 1976; Hepler et al., 1976; Merritt et al., 1980), lasting up to 4 hours post-administration (Hepler and Petrus, 1976). Intravenous administration of cannabinoids also reduced IOP within 30 minutes (Cooler and Gregg, 1977; Purnell and Gregg, 1975).

[0005] Many of these studies, however, reported that systemic administration of centrally acting camiabinoids resulted in significant psychiatric and minor cardiovascular side-effects (Merritt et al., 1980). Flach (2002) reported that 8 out of 9 patients dropped out of a 9-month THC program within 8--20 weeks due to tolerance and toxicity-related phenomena.

[0006] In addition, antagonizing central CBi receptors is associated with CNS-mediated neuropsychiatric side effects such as low mood, reduced joy, anxiety, depression, and suicidal ideology, due to the important role that CBi receptors play in the brain’s reward system (Panagis G., Mackey B., Vlachou S., Front. Psychiatry. 2014;5:92).

[0007] To overcome the potential issues regarding psychiatric adverse events (AEs), the use of a peripherally restricted cannabinoid, such as disclosed herein, may avoid these complications, while still exerting a beneficial effect on eye disorders.SUMMARY OF THE DISCLOSURE

[0008] A first aspect of the present disclosure are methods for treating eye disorders in a subject needing treatment using a peripherally restricted cannabinoid.

[0009] In an embodiment of the first aspect, the peripherally restricted cannabinoid for treating eye disorders is Compound I having the following formula:Compound I or a pharmaceutically acceptable salt thereof.

[0010] In another embodiment, disclosed herein are methods for reducing intraocular pressure in a subject having elevated IOP, wherein the method involves administering to the subject a therapeutically effective amount of Compound 1.

[0011] In some embodiments, the administration of Compound I reduces the intraocular pressure as compared to the intraocular pressure before administration.

[0012] In yet other embodiments, the administration of Compound I reduces the intraocular pressure by about 0.1 mmHg to about 15 mmHg as compared to the intraocular pressure before administration.

[0013] In other embodiments, the administration of Compound I reduces the intraocular pressure in a subject that has glaucoma.

[0014] In still other embodiments, Compound I is administered orally.

[0015] In some embodiments, Compound l is a pharmaceutical composition useful for treating an eye disorder.

[0016] In some embodiments, Compound I is a pharmaceutical composition useful for treating increased intraocular pressure including intraocular pressure associated with glaucoma.

[0017] In yet other embodiments, the administration of Compound I to a subject with an eye disorder, such as, without limitation, elevated intraocular pressure and / or glaucoma protects retinal ganglion cells and / or attenuates optic nerve damage.

[0018] In some embodiments, the eye disorder is associated systemic disorders associated with glaucoma, such as, without limitation, acute retinal necrosis Behcet disease, HLA-B27-related acute anterior uveitis, juvenile idiopathic arthritis- associated uveitis, sarcoidosis, Vogt-Koyanagi-Harada syndrome, acquired immunodeficiency syndrome, cytomegalovirus retinitis, congenital rubella,disseminated meningococcemia, Hansen disease (leprosy), herpes virus-associated uveitis, Lyme disease, onchocerciasis, syphilis, toxocariasis, and toxoplasmosis.

[0019] In other embodiments, the eye disorder is caused or associated with one or more of the following systemic diseases, such as, without limitation, Axenfeld- Rieger syndrome, Crouzon syndrome (craniofacial dysostosis), Cushing disease, Bing- Neel syndrome, Trisomy 21, Trisomy 16-18 (Edward syndrome), Trisomy 13-15 (Patau syndrome), Turner syndrome, 9p syndrome, Cockayne syndrome, Cretinism (juvenile hypothyroidism), Cystinosis, Dental-ocular-cutaneous syndrome, Diabetes, Diamond- Blackfan syndrome, Ehlers-Danlos syndrome, Familial histiocytic demiatoarthritis syndrome, Fetal alcohol syndrome, Gorlin-Goltz syndrome, Hallermann-Streiff syndrome (oculomandibulofacial dyscephaly), Homocystinuria, Kartagener syndrome, Kimmelstiel- Wilson syndrome, Klinefelter syndrome, Klippel-Trenaunay-Weber syndrome, Krabbe disease, Krause syndrome, Lowe (oculocerebrorenal) syndrome, Marfan syndrome, Meyer-Schwickerath-Weyers syndrome, Miller (Wilms aniridia) syndrome, Mucopolysaccharidosis, neurofibromatosis (Von Recklinghausen), nevus of ota (oculodermal melanocytosis), PHACES syndrome, Pierre Robin syndrome, Prader- Willi syndrome, Rubinstein-Taybi syndrome, Sickling disorders, Stickler syndrome, Sturge-Weber syndrome, Treacher Collins syndrome, Ullrich syndrome (dyscraniopygophalangy), Von Hippel-Lindau disease, Waardenburg syndrome, Wagner syndrome, Weber-Christian disease, Weil-Marchesani syndrome, X-linked mental retardation syndrome, and Zellweger (cerebrohepatorenal) syndrome.BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Fig. 1- shows concentrations of Compound 1 and / or its metabolites in various tissues in male rats at different times after oral administration.DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] Drugs that are currently used or being investigated in clinical research to treat glaucoma may reduce IOP in some patients. However, IOP is only one factor in the pathogenesis of glaucoma. Glaucoma is also characterized by degeneration of the retinal ganglion cells (RGCs) and optic nerve damage. Currently used drug classes are not known to be neuroprotective, i.e., they may reduce IOP, but they haveno protective effect on RGCs nor do they attenuate optic nerve damage (Zhang et al., 2012). While cannabinoids can reduce IOP, they also have anti-oxidative and antiinflammatory effects (Klein et al., 1998; Marsicano et al., 2002). Camiabinoids are thought to protect RGCs via inhibition of glutamate (excitatory neurotransmitter), endothelin- 1 (vasoconstrictor), and nitric oxide (oxidative stress mediator), which are compounds known to be involved in the pathology of glaucoma (Samy et al., 1994). For example, long-term administration of A-9-THC in rats reduces IOP and attenuates RGC cell death by up to 75% (Schwitzer et al., 2015). A-9-THC -mediated inhibition of endothelin- 1 in humans facilitates adaptive blood perfusion of the optic nerve whilst also increasing retinal perfusion (Green et al., 1978; Hommer et al., 2020). AEA- mediated activation of retinal and central CBi and CB2 receptors protects RGCs via inhibition of nitric oxide and inflammatory cytokines associated with oxidative stress (Krishnan & Chatterjee, 2015).

[0022] Increased IOP can be caused by systemic disorders associated with glaucoma, such as, without limitation, acute retinal necrosis Behpet disease, HLA-B27 related acute anterior uveitis, juvenile idiopathic arthritis-associated uveitis, sarcoidosis, Vogt-Koyanagi-Harada syndrome, acquired immunodeficiency syndrome, cytomegalovirus retinitis, congenital rubella, disseminated meningococcemia, Hansen disease (leprosy), herpes virus-associated uveitis, lyme disease, onchocerciasis, syphilis, toxocariasis, and toxoplasmosis.

[0023] Increased IOP can also be associated with systemic diseases, such as, without limitation, Axenfeld-Rieger syndrome, Crouzon syndrome (craniofacial dysostosis), Cushing disease, Bing-Neel syndrome, Trisomy 21, Trisomy 16-18 (Edward syndrome), Trisomy 13-15 (Patau syndrome), Turner syndrome, 9p syndrome, Cockayne syndrome, Cretinism (juvenile hypothyroidism), Cystinosis, Dental-ocular- cutaneous syndrome, Diabetes, Diamond-Blackfan syndrome, Ehlers-Danlos syndrome, Familial histiocytic dermatoarthritis syndrome, Fetal alcohol syndrome, Gorlin-Goltz syndrome, Hallermann-Streiff syndrome (oculomandibulofacial dyscephaly), Homocystinuria, Kartagener syndrome, Kimmelstiel-Wilson syndrome, Klinefelter syndrome, Klippel-Trenaunay-Weber syndrome, Krabbe disease, Krause syndrome, Lowe (oculocerebrorenal) syndrome, Marfan syndrome, Meyer- Schwickerath-Weyers syndrome, Miller (Wilms aniridia) syndrome,Mucopolysaccharidosis, neurofibromatosis (Von Recklinghausen), nevus of ota(oculodermal melanocytosis), PHACES syndrome, Pierre Robin syndrome, Prader- Willi syndrome, Rubinstein-Taybi syndrome, Sickling disorders, Stickler syndrome, Sturge-Weber syndrome, Treacher Collins syndrome, Ullrich syndrome (dyscraniopygophalangy), Von Hippel-Lindau disease, Waardenburg syndrome, Wagner syndrome, Weber-Christian disease, Weil-Marchesani syndrome, X-linked mental retardation syndrome, and Zellweger (cerebrohepatorenal) syndrome.

[0024] Disclosed herein is the use of a synthetic cannabinoid N-(2-(tert- butyl)- 1 -((4,4-difluorocyclohexyl)methyl)- 1 H-benzimidazol-5 -yl)ethanesulfonamide including salts, and pharmaceutical formulations thereof to treat eye disorders, such as, without limitation, increased intraocular pressure (IOP). The structure of N-(2-(tert- butyl)- 1 -((4,4-difluorocyclohexy l)methyl)- 1 H-benzimidazol-5 -yl)ethanesulfonamide (Compound I) is shown below:Compound 1

[0025] Compound I has demonstrated potential in cancer anorexia (appetite and weight loss) as well as cancer-induced muscle degeneration (cachexia) and may have utility in treating other forms of anorexia through its dual CBi and CEk receptor agonism. CBi receptor activation has been shown to increase food intake, alter adipokine and satiety hormone levels, alter taste sensation, decrease lipolysis (fat break down), and increase lipogenesis (fat generation).

[0026] As disclosed herein, the distribution and retention of radioactivity of [14c]Compound 1 and / or its metabolites after single oral administration was studied in pigmented male rats. As described in Example I, following administration to male rats, [14c]Compound I and / or its metabolites were widely distributed in the body. Maximalconcentration of radioactivity was reached after 4 hours in all tissues, except renal outer medulla and epididymis where the peak was reached after 1 and 24 hours, respectively (see Table 2 in Example 1). The highest tissue levels of radioactivity were found in liver, uvea+RPE, kidney, and adrenal. Throughout the study most of the tissues had concentrations in the same range or higher than those in the blood. The half-life in blood was 10 hours. Compound I and / or its metabolites were excreted primarily via bile and at 6 days after the administration most of the radioactivity was eliminated. After 21 days no radioactivity was observed in the body.

[0027] As disclosed in Example 2, Compound I is tested in mammals, such as humans to determine if it can reduce IOP through its CBi receptor agonism and neuroprotection through its CB2 receptor agonism.

[0028] As used herein, the terms "administer," "administering", "administration," and the like refer to the methods that may be used to enable delivery of compounds or compositions to the desired site of biological action. These methods include, but are not limited to oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), topical, and rectal administration. Those of skill in the art are familiar with administration techniques that can be employed with the compounds and methods described herein.

[0029] In some embodiments, the compounds and compositions described herein are administered orally.

[0030] The terms “coadministration” or “administration in conjunction with” or “cotreatment” when used in reference to the coadministration of a first compound (e.g., Compound I) and a second compound indicates that the first compound and the second compound are administered so that there is at least some chronological overlap in the biological activity of first compound and the second compound in the organism to which they are administered. Coadministration can be simultaneous administration or sequential administration. In sequential administration there may even be some substantial delay (e.g., minutes or even hours) between administration of the first compound and the second compound as long as their biological activities overlap. In certain embodiments the coadministration is over a time frame that permits the first compound and second compound to produce an enhanced effect (e.g.,therapeutic or prophylactic effect) on the organism. In certain embodiments the enhanced effect is a synergistic effect.

[0031] The terms “effective amount” or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered, which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result includes reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an “effective amount” for therapeutic uses is the amount of the composition comprising a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms. An appropriate “effective” amount in any individual case is optionally determined using techniques, such as a dose escalation study.

[0032] The term “pharmaceutical combination” as used herein, means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term “fixed combination” means that the active ingredients, e.g., a compound described herein, or a pharmaceutically acceptable salt thereof, and a co-agent, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the active ingredients, e.g., a compound described herein, or a pharmaceutically acceptable salt thereof, and a co-agent, are administered to a patient as separate entities either simultaneously, concurrently, or sequentially with no specific intervening time limits, wherein such administration provides effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g., the administration of three or more active ingredients.

[0033] The term “subject” or “patient” encompasses mammals. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, and guinea pigs, and the like. In one aspect, the mammal is a human.

[0034] The terms “treat,” “treating” or “treatment,” as used herein, include alleviating, abating, or ameliorating at least one symptom of a disease or condition, preventing additional symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition either prophylactically and / or therapeutically.

[0035] The compounds used in the method of the present invention may be prepared by techniques well known in organic synthesis and familiar to a practitioner ordinarily skilled in the art. However, these may not be the only means by which to synthesize or obtain the desired compounds.

[0036] The compounds used in the method of the present invention may be prepared by techniques described in Vogel's Textbook of Practical Organic Chemistry, A. I. Vogel, A. R. Tatchell, B. S. Furnis, A. J. Hannaford, P. W. G. Smith, (Prentice Hall) Sth Edition (1996), March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Michael B. Smith, Jerry March, (Wiley-Interscience) 5th Edition (2007), and references therein, which are incorporated by reference herein. However, these may not be the only means by which to synthesize or obtain the desired compounds.

[0037] In other embodiments, the compound used in the present invention can be prepared, without limitation, as described in US Patent No. 7,550,495; Example 14, column 46 or International application no. PCT / GB2021 / 053200.Pharmaceutical Compositions:

[0038] The term “pharmaceutically acceptable salt” in reference to Compound I refers to a salt of Compound I, which does not cause significant irritation to a mammal to which it is administered and does not substantially abrogate the biological activity and properties of the compound. Handbook of Pharmaceutical Salts: Properties, Selection and Use. International Union of Pure and Applied Chemistry, Wiley-VCH 2002. S.M. Berge, L.D. Bighley, D.C. Monkhouse, J. Pharm. Sci. 1977, 66, 1-19. P. H. Stahl and C. G. Wermuth, editors, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim / Zurich:Wiley-VCH / VHCA, 2002. In some embodiments, pharmaceutical salts typically are more soluble and more rapidly solubleill stomach and intestinal juices than non- ionic species and so are useful in solid dosage forms. Furthermore, because their solubility often is a function of pH, selective dissolution in one or another part of the digestive tract is possible, and this capability can be manipulated as one aspect of delayed and sustained release behaviors. Also, because the salt-forming molecule can be in equilibrium with a neutral form, passage through biological membranes can be adjusted.

[0039] In some embodiments, pharmaceutically acceptable salts are generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base. The term may be used in reference to any compound of the present invention. Representative salts include the following salts: Acetate, Benzenesulfonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate, Borate, Bromide, Calcium Edetate, Camsylate, Carbonate, Chloride, Clavulanate, Citrate, Dihydrochloride, Edetate, Edisylate, Estolate, Esylate, Fumarate, Gluceptate, Gluconate, Glutamate, Glycollylarsanilate, Hexylresorcinate, Hydrabamine, Hydrobromide, Hydrochloride, Hydroxynaphthoate, Iodide, Isethionate, Lactate, Lactobionate, Laurate, Malate, Maleate, Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate, Monopotassium Maleate, Mucate, Napsylate, Nitrate, N-methylglucamine, Oxalate, Pamoate (Embonate), Palmitate, Pantothenate, Phosphate / diphosphate, Polygalacturonate, Potassium, Salicylate, Sodium, Stearate, Subacetate, Succinate, Tamiate, Tartrate, Teoclate, Tosylate, Triethiodide, Trimethylammonium, and Valerate. When an acidic substituent is present, such as -CO2H, there can be formed the ammonium, morpholinium, sodium, potassium, barium, calcium salt, and the like for use as the dosage form. When a basic group is present, such as amino, or a basic heteroaryl radical, such as pyridyl, there can be formed an acidic salt, such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate, methanesulfonate, ethanesulfonate, picrate, and the like, and include acids related to the pharmaceutically acceptable salts listed in Stephen M. Berge, et al., Journal of Pharmaceutical Sciences, Vol. 66(1), pp. 1-19 (1977).

[0040] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. Pharmaceutical compositions areformulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active compounds into preparations that are used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions described herein is found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton. Pennsylvania 1975; Liberman. H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), herein incorporated by reference for such disclosure.

[0041] In some embodiments, the compounds are formulated as described in International application no. PCT / GB2021 / 053200.

[0042] In some embodiments of the invention, Compound I is included within a pharmaceutical composition. As used herein, the term 'pharmaceutical composition” refers to a liquid or solid composition, preferably solid, that contains a pharmaceutically active ingredient, i.e., Compound I and at least a carrier, where none of the ingredients is generally biologically undesirable at the administered quantities.ExamplesExample 1 - Eye Data Compound I:Study Design:

[0043] Route of administration- Oral, by gavage.

[0044] Frequency of administration- single dose.

[0045] Dose levels are specified in Table 1.

[0046] Animal usage- one rat was use used for each time point.Table 1 Details of animals, dosages, and dose volumesAnimals Route of Dose levels Dose Volume Time after administration pmol.kg1MBq.kg-1mL.kg-1administration(1 mg / kg)Male Oral 2.5 5 6 5 minLister 15 minHooded 1 hour4 hours24 hours2 days6 days21 daysTest item formulations:

[0047] Specific radioactivity 2.05 MBq / pmo

[0048] Radiochemical purity 99.3%

[0049] Test item formulation Compound I was formulated for dosing as solution water purified, adjusted to pH 2 for oral administration.

[0050] Formulation concentrations The obtained concentrations were 0.40 pmol / mL and.0.84 MBq / mL.

[0051] Radiochemical purity The radiochemical purity of the test compound in the formulation was about 98.9%.Animals:

[0052] Strain Wistar Hannover Galas (albino) and Lister hooded (pigmented)

[0053] Number of animals 13

[0054] Age range Males, 6 to 8 weeks on days of dosing.

[0055] Body weight range Males 207 g to 219 g at administration.

[0056] All animals were observed daily during the study.

[0057] Animals were observed for changes in appearance and behavior, for signs of ill health and for mortality.

[0058] Body weight Body weights of all animals were recorded a few days after arrival and accurately, prior to dosing.Autoradiography:Euthanasia and sectioning:

[0059] The animals were killed with enflurane (Efrane®, Abbott Laboratories, USA) and frozen in acetone, cooled to -70°C with solid CO2.After removal of limbs and tail, the carcass was embedded, with left lateral side uppermost, in a 2.5% aqueous solution of carboxymethyl cellulose and frozen for at least 10 minutes in acetone at -70°C. The animal blocks were stored at -20°C until sectioning.

[0060] At sectioning each block was mounted in a Leica CM3600 Cryostat (Leica Microsystems AB, Sweden) maintained at approximately -20°C. Sagittal wholebody sections (20 pm) were obtained at various levels through the carcass to include all major organs and tissues of interest. The sections were mounted on type 810 tape (Minnesota 3M, USA) and numbered consecutively with radioactive ink. From the animals killed 7 and 21 days after administration, a couple of sections were mounted on type 6890 tape (3M, USA), which were not extracted.

[0061] All sections were dried at -20°C for >3 days prior to exposure on phosphor imaging plates. Radioactive calibration standards prepared from the test formulation were frozen and sectioned in an identical manner.Preparation of standards:

[0062] For calibration of the phosphor imaging system a series of radioactive nuclide standards was used. This standard series, made in an aqueous solution of gelatine (100 mg / mL), consisted of 7 dilutions from 0.07 to 19.18 kBq / g, corresponding to 0.03 to 9.11 nmol / g. For the purpose of quantification, it was assumed that all tissues had similar density and quench characteristics as the gelatine solution. Phosphor imaging analysis:

[0063] Sections were chosen for phosphor imaging to best represent the tissues and organs of interest. Together with two sets of calibration standards the sections were placed on phosphor imaging plates, which had been pre-covered with a thin plastic film. The imaging plates were exposed for 6 hours to 3 days at +4 to +12°C, enclosed in light tight cassettes in a lead shielding box to protect from environmental radiation.

[0064] Following exposure, the imaging plates were scanned at a pixel size of 50 pm using BAS 2500 (FujiFilm Sverige AB. Sweden). The tissues and organs ofinterest were quantified using AIDA (Raytest, Germany). The quantification limit is defined as twice the background value of the image plate and varied between the plates from 0.029 to 0.065 nmol / g.

[0065] The various tissues and organs were identified either on the autoradiograms or on the corresponding tissue sections.Calculations of terminal half-lives:

[0066] The terminal half-life of total radioactivity was estimated for some selected tissues. The half-life was calculated as ln2 / k, where k is the coefficient slope in a graph plotted with In transformed values of concentration. The last three measurable time-points were generally used in the calculations. For liver and uvea+RPE the four last time point were used and for blood only 2 time points were used (Table 3).RESULTS:Clinical observations:

[0067] The animals showed no signs of being affected by the treatment. Distribution of radioactivity:

[0068] The radioactivity' in the sections represents both the administered test compound and its metabolites. The quantitative evaluation is given in Table 2 and in Fig. 1. Terminal half-lives are presented in Table 3. The term uvea + RPE (retinal pigment epithelium) used in this study represents all melanin containing structures of the eyeTable 2- Male Lister Hooded rats, single oral dose 2.5 pmol / kg, tissue concentration Compound I and / or its metabolitesTissue Times after administration5 min 15 min 1 h 4 h 24 h 2 d 6 d 21 dAdrenal ni fl 0.33 (5) 1.0 (8) 0.18 (5) 0.072 (17) blqBile 1.0(3) 5.1(6) 6.1(10) 12(4) 4.1(26) 0.27(17)Bone marrow ni 0.16(9) 0.13(2) 0.33(8) 0.082(1) ni niBrain blq blq blq blq blqBrownfat Ni 0.14(1) 0.28(0) 0.71(1) 0.18(5) 0.073(0) niEpididymis blq blq 0.11(3) 0.5(11) 1.2(0) 0.95(1) blqGastric mucosa fl fl fl fl 0.40(0) 0.042(13) blqHeart blood blq 0.086(2) 0.10(0) 0.27(1) 0.072(2) blqLens ni blq blq blq blq blqLiver 1.1(2) 1.4(2) 1.0(1) 2.8(2) 1.3(8) 1.1(1) 0.034(0) blqLung blq 0.17(3) 0.13(2) 0.35(3) 0.085(5) blqLymph node ni 0.11(5) 0.12(5) 0.26(7) ni niMyocardium 0.049(6) 0.30(3) 0.25(1) 0.59(0) 0.15(8) 0.043(5) blqPancreas ni 0.33(12) 0.43(0) 0.92(3) 0.18(2) 0.048(10) blqParotid salivary ni 0.27(0) 0.23(5) 0.64(-) 0.11(5) blq blq gladPineal body ni 0.14(5) 0.19(7) 0.37(24) 0.086(7) niPituitary gland ni 0.13(1) 0.18(12) 0.63(10) 0.15(32) blqProstate gland ni 0.068(25) 0.16(18) 0.68(5) 0.51(7) 0.092(19) blqRenal cortex 0.079(6) 0.33(2) 0.33(10) 0.87(5) 0.21(5) 0.070(2) blqRenal medulla 0.047(6) 0.28(5) 0.72(6) 0.54(0) 0.11(3) 0.035(18) Ni(inner)Renal medulla 0.070(-) 0.80(0) 0.41(6) 1.7(4) 0.38(6) 0.089(2) ni(outer)Skeletal muscle blq 0.096(8) 0.12(2) 0.33(2) 0.065(2) blq blqSkin, non- blq 0.10(3) 0.14(10) 0.32(1) 0.083(1) 0.048(7) blq pigmentedSkin, pigmented ni 0.086(2) 0.12(5) 0.27(2) 0.062(7) blq niSpinal cord big blq blq blqSpleen 0.052(1) 0.19(21) 0.17(4) 0.49(6) 0.097(0) 0.035(14) blqSubmandibular ni 0.20(11) 0.20(6) 0.51(1) 0.11(6) 0.038(13) ni salivary glandTestis big blq blq 0.12(1) 0.050(20) blqThymus blq 0.066(5) 0.10(3) 0.26(1) 0.063(0) blqThyroid gland ni 0.18(1) 0.23(1) 0.50(0) 0.13(10) ni niUrine ni 0.089(3) 0.58(24) 1.6(11) 1.4(15) 0.17(11) niUvea+RPE ni 0.17(14) 0.60(1) 2.4(4) 1.1(2) 0.23(4) 0.031(46) niLimit of 0.035 0.037 0.034 0.034 0.033 0.034 0.029 0.030 quantificationValues are expressed in nmol / g and coefficient of variation (%) blq below limit of detection ni not identified for quantification fl not measure due to flare from adjacent tissueSUBSTITUTE SHEET (RULE 26)Table 3 Male Lister Hooded rats, terminal half-lives of total radioactivity' in tissues after single oral administration of [14c] Compound I _Tissue Half-life (hours)3RevalueAdrenal 12 0.94Heart blood 10 1Liver 22 0.98Prostate gland 20 0.51Renal medulla outer 10 0.99Uvea+RPE 23 0.94Distribution of radioactivity in male pigmented rats after oral administration:

[0069] At 5 minutes after oral administration distribution of radioactivity' was limited to a few tissues and organs. Most of the radioactivity was found in the stomach. Other tissues with measurable concentrations were the liver, renal cortex and outer medulla, spleen, myocardium, and in the bile (in descending order). The concentrations in the brain, spinal cord, and blood w ere below' limit of quantification throughout the study. Radioactivity was found in the small intestine. No radioactivity was found in the lens at any time point.

[0070] At 15 minutes the overall level of radioactivity in the body was increased. Highest tissue concentration was found in bile content, liver, renal outer medulla, cortex, and the myocardium. The concentration in blood was above quantification limit but lower or at the same level as all measurable tissues.

[0071] At 1 hour the general level was somewhat increased, however a few tissues had decreased concentrations. The highest concentration was found in the bile, liver, renal inner medulla, uvea + RPE, and urine. A great amount of radioactivity' was present in the stomach content and a smaller amount in the small intestine.

[0072] At 4 hours the maximal concentration was reached in all tissues except for the renal inner medulla. Again, the highest concentrations were found in the bile, liver, uvea + RPE, renal outer medulla, and pancreas. Still a great amount of radioactivity was present in the stomach content. Radioactivity was also noticed in the small intestine, caecum, and colon wall.

[0073] At 24 hours all tissue concentrations were markedly reduced, but still measurable in the same tissues as after 4 hours. Highest concentrations were foundin bile, liver, urine, uvea + RPE, and prostate gland. Most tissues had concentrations in the same range or higher than the level in blood. Highest level in the gastro-intestinal tract was now in caecum and colon content. Radioactivity was noticed in skin around the head.

[0074] At 2 days after the administration the concentrations were further decreased and were now below quantification limit in one third of the tissues. Highest concentrations were seen in the liver, bile, uvea + RPE, urine, and the prostate gland. The level in blood was now below quantification limit.

[0075] At 6 days the only tissues with measurable levels of radioactivity were the liver and uvea+RPE, just at quantification limit.

[0076] At 21 days no measurable radioactivity could be detected in the body RESULTS:Distribution of radioactivity:

[0077] The quantitative evaluation is given in Figure 1 which shows the concentrations of Compound I and / or its metabolites in various tissues at different times after oral administration of 2.5 pmol / kg to male pigmented rats. The term uvea + RPE (retinal pigment epithelium) used in this study represents all melanin containing structures of the eye.

[0078] Distribution of radioactivity in male pigmented rats after oral administration. At 5 minutes after oral administration distribution of radioactivity was limited to a few tissues and organs (data not shown). Most of the radioactivity was found in the stomach. Other tissues with measurable concentrations were the liver, renal cortex and outer medulla, spleen, myocardium, and in the bile (in descending order). The concentrations in the brain, spinal cord, and blood were below limit of quantification throughout the study. Radioactivity was found in the small intestine. No radioactivity was found in the lens at any time point.

[0079] At 15 minutes the overall level of radioactivity in the body was increased. Highest tissue concentration was found in bile content, liver, renal outer medulla, cortex, and the myocardium (data not shown). The concentration in blood was now above quantification limit but lower or at the same level as all measurable tissues.

[0080] At 1 hour the general level was somewhat increased, however a few tissues had decreased concentrations (data not shown). Highest concentration wasfound in the bile, liver, renal inner medulla, uvea + RPE, and urine. A great amount of radioactivity was present in the stomach content and a smaller amount in the small intestine.

[0081] At 4 hours the maximal concentration was reached in all tissues except for the renal inner medulla (data not shown). The highest concentrations were found in the bile, liver, uvea + RPE, renal outer medulla, and pancreas. Still a great amount of radioactivity was present in the stomach content. Radioactivity was also noticed in the small intestine, caecum, and colon wall.

[0082] At 24 hours all tissue concentrations were markedly reduced, but still measurable in the same tissues as after 4 hours (data not shown). Highest concentrations were found in bile, liver, urine, uvea + RPE, and prostate gland. Most tissues had concentrations in the same range or higher than the level in blood. Highest level in the gastro-intestinal tract was now in caecum and colon content. Radioactivity was noticed in skin around the head.Example 2- Study Design:

[0083] A randomized, cross-over, single center pilot study will be performed. Ten patients, over the age of 40 years, with ocular hypertension or mild primary open angle glaucoma will be recruited. Mild glaucoma will be defined as a visual field loss less than 6 dB in the standard 24-2 Humphrey visual field test. The subject's IOP at randomization must fall between 20 and 35 mmHg inclusive, in either an untreated or washed-out state. All IOP measurements will be taken using Goldmann applanation tonometry (GAT).Exclusion criteria:

[0084] Any other clinically significant disease in either eye, prior incisional or laser glaucoma surgery, cataract surgery in the last 6 months, significant systemic disease, use of cannabinoids for other purposes within a year of the study visit, currently pregnant, considering pregnancy or not using reliable birth control, use of systemic beta-blockers, use of aspirin or other NSAID.Study processes:

[0085] If subjects are currently using lOP-lowering medication, a 1 -month wash-out period will commence, after which a baseline visit will be scheduled. During the washout period, additional visits can be conducted, at the discretion of theinvestigator, to ensure continued safety, particularly for subjects considered at risk of an unacceptably high IOP spike during washout. If during one of these visits it is found that the subject’s IOP increases to an unsafe level (at the investigators’ discretion), ocular hypotensive medication will be reintroduced, and they will be followed for safety until any AEs are resolved or stabilized. They will then be withdrawn from the study. Following washout, the eligibility of subjects will be confirmed at the baseline visit. Threshold IOP value will be determined based on the investigators’ clinical judgement. For subjects not requiring wash out, the baseline assessments will be carried out as part of the first screening visit and a separate baseline visit will not be necessary.

[0086] In a randomly chosen order, a daily dose of Compound I (650 microgram) or placebo will be given for two weeks. Values for the main outcome measure (IOP) and secondary measures (heart rate, blood pressure [BP]) will be taken at all research visits. Subject’s weight will also be obtained at each visit. Determination of IOP, heart rate, and BP will always be made at the same time of the day (+ / - 2 hours) due to diurnal variation.

[0087] To determine the IOP the average of two GAT readings of IOP will be used, with a third reading to be taken if the difference between the first two readings is greater than 3 mmHg.

[0088] Subject tolerability of the treatment will be assessed at all visits following randomization. At each visit. IOP will be assessed and compared with the threshold IOP limit value previously set by the investigator. If this value is reached, topical hypotensive medications will be initiated, starting with prostaglandin analogues, then beta-blocker and other medications according to standard of care.

[0089] Treatment modification will be allowed for safety reasons if IOP becomes uncontrolled after treatment. There will be no reintroduction of medication unless, in the opinion of the investigator, there is immediate risk to the health of the participant.Safety considerations:

[0090] The acute toxicity of Compound I is very low. Unwanted effects occur not uncommonly but are generally w ell tolerated by patients. Most common AEsobserved in clinical studies so far are postural dizziness, nausea, hypotension, and headache, all mild to moderate in severity. The incidence of AEs is low.

[0091] Withdrawing current topical medication in patients with ocular hypertension and mild glaucoma for a period of weeks is considered to be safe and is the current standard practice in clinical research. Early glaucoma is a slow chronic disease, and no risks to visual function are anticipated.

[0092] Safety will be evaluated throughout the study consisting of recording of all AEs. An independent medical monitor will be responsible for overseeing safety and for evaluating all AEs reported to the sponsor. AEs and serious AEs are defined according to the International Conference on Harmonization E6 (R2) and International Organization for Standardization 14155:2011(E) definition.Statistical analyses and sample size calculation:

[0093] In the Early Manifest Glaucoma Trial (Leske et al., 1999), the standard deviation (SD) of untreated IOP over time was 2.02 mmHg and 1.78 mmHg in the untreated and treated arms, respectively. We have used a SD of 2 mmHg for our sample size estimation.

[0094] With a sample size in each sequence group of 5, (a total sample size of 10) a 2 x 2 crossover design will have 90% power to detect a difference in mean IOP of 2.4 mmHg, assuming that the Crossover ANOVA sqrt (MSE) is 1.414 (the Standard deviation of differences, o, is 2) using a two-group t-test (Crossover ANOVA) with a 0.05 two-sided significance level.

[0095] A difference above 2 mmHg in IOP is considered to be clinically important by European glaucoma experts (EGS consensus survey).

[0096] At baseline visit we will define a study eye for the main analysis, i.e., the eye with highest IOP. In case of both eyes having similar IOP the study eye will be the one with more advanced disease.

[0097] The examples and embodiments described herein are for illustrative purposes only and various modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application and scope of the appended claims.

Claims

WHAT IS CLAIMED IS:

1. A method for treating an eye disorder in a subject in need of treatment, wherein the method comprises administering to the subject a therapeutically effective amount of Compound I having the formula:Compound I or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the subject treated has an eye disorder associated with increased intraocular pressure3. The method of claim 2, wherein the administration of Compound I reduces the intraocular pressure as compared to the intraocular pressure before administration.

4. The method of claim 2. wherein the administration of Compound I reduces the intraocular pressure by about 0.1 mmHg to about 15 mmHg as compared to the intraocular pressure before administration.

5. A method for protecting retinal ganglion cells and / or preventing optic nerve damage in a subject having an eye disorder, wherein the method comprises administering to the subject a therapeutically effective amount of Compound I.

6. The method of any one of claims 1-5, wherein Compound I is administered orally.

7. The method of any one of claims 1-6. wherein the subject has glaucoma.

8. The method of any one of claims 1-7. wherein Compound 1 is a pharmaceutical composition.

9. The method of any one of claims 1-8, wherein the eye disorder is associated with systemic disorders associated with glaucoma selected from one or more of acute retinal necrosis Behcet disease, HLA-B27-related acute anterior uveitis, juvenile idiopathic arthritis-associated uveitis, sarcoidosis, Vogt-Koyanagi -Harada syndrome, acquired immunodeficiency syndrome, cytomegalovirus retinitis, congenital rubella, disseminated meningococcemia. Hansen disease (leprosy), herpes virus- associated uveitis, lyme disease, onchocerciasis, syphilis, toxocariasis, and toxoplasmosis.

10. The method of any one of claims 1-8, wherein the eye disorder is associated with systemic diseases selected from one or more of Axenfeld-Rieger syndrome, Crouzon syndrome (craniofacial dysostosis), Cushing disease, Bing-Neel syndrome. Trisomy 21, Trisomy 16-18 (Edward syndrome). Trisomy 13-15 (Patau syndrome), Turner syndrome, 9p syndrome, Cockayne syndrome, Cretinism (juvenile hypothyroidism), Cystinosis, Dental-ocular-cutaneous syndrome, Diabetes, Diamond- Blackfan syndrome, Ehlers-Danlos syndrome. Familial histiocytic dermatoarthritis syndrome, Fetal alcohol syndrome, Gorlin-Goltz syndrome, Hallermann-Streiff syndrome (oculomandibulofacial dyscephaly), Homocystinuria, Kartagener syndrome, Kimmelstiel-Wilson syndrome, Klinefelter syndrome, Klippel-Trenaunay-Weber syndrome, Krabbe disease, Krause syndrome, Lowe (oculocerebrorenal) syndrome, Marfan syndrome. Meyer-Schwickerath-Weyers syndrome, Miller (Wilms aniridia) syndrome, Mucopolysaccharidosis, neurofibromatosis (Von Recklinghausen), nevus of ota (oculodermal melanocytosis), PHACES syndrome, Pierre Robin syndrome, Prader- Willi syndrome, Rubinstein-Taybi syndrome, Sickling disorders, Stickler syndrome, Sturge-Weber syndrome, Treacher Collins syndrome. Ullrich syndrome (dyscraniopygophalangy). Von Hippel-Lindau disease, Waardenburg syndrome, Wagner syndrome, Weber-Christian disease, Weil-Marchesani syndrome, X-linked mental retardation syndrome, and Zellweger (cerebrohepatorenal) syndrome.

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