Compositions and methods for use of cannabinol compounds in neuroprotection

Abstract

Provided herein are methods and compositions for neuroprotection. The neuroprotective composition can be or include 4-chlorobutyl-cannabinol (4-chlorobutyl-CBN) or a derivative thereof and / or a pharmaceutically acceptable salt thereof. The neuroprotective composition can be used in the treatment of a neurodegenerative disease. The neuroprotective composition can be used to protect retinal neurons from degeneration in a subject in need thereof, such as for treatment of age-related macular degeneration.

Description

Attorney Docket No.120754.00100 COMPOSITIONS AND METHODS FOR USE OF CANNABINOL COMPOUNDS IN NEUROPROTECTION BACKGROUND

[0001] Neurodegeneration is a phenomenon underlying a wide array of different diseases of the central and peripheral nervous system. Neurodegeneration includes neuronal atrophy, axonal degeneration (e.g., Wallerian and / or Wallerian-like degeneration), and induction of necrotic or programmed mechanisms of cell death. Different types of programmed cell death such as apoptosis, autophagy, pyroptosis, and oncosis have been demonstrated in neurons. Stimuli such as physical injury, oxidative stress, excitotoxicity, mitochondrial dysfunction, inflammation, iron accumulation, and protein aggregation have been shown to contribute to mechanisms of neurodegeneration.

[0002] Eye diseases associated with neural degeneration include age-related macular degeneration (AMD). AMD affects about 14-24% of people aged 65 to 74 and about 35% of people over 75 around the world, and results in vision impairment or loss in the center of the visual field (the macula), because of damage to the retina and / or associated neurons. AMD is a cause of vision loss and potential blindness in people over 50 years of age. The two principal forms of AMD are atrophic (non-exudative or “dry”) AMD and neovascular (exudative or “wet”) AMD. Atrophic AMD is characterized by geographic atrophy (GA) at the center of the macula in the advanced stage of AMD, and vision can slowly deteriorate over many years due to the loss of photoreceptors and development of GA. Neovascular AMD is a more severe form of AMD and is characterized by neovascularization (e.g., choroidal neovascularization) in the advanced stage of AMD, which can rapidly lead to blindness. Neovascular AMD affects more than 30 million patients worldwide and is a leading cause of vision loss in people aged 60 years or older. If AMD remains untreated, patients are likely to lose central vision in the affected eye within 24 months of disease onset. About 90% of AMD patients have the dry form, and about 10% develop neovascular AMD.

[0003] Neuroprotection is an effect that can provide salvage or recovery of the nervous system, its cells, structure, and / or function, or resistance to neurodegenerative stimuli. Neuroprotective compositions may find use in treating a variety of diseases that cause or result in neurodegeneration, such as age-related macular degeneration, or in mitigating symptoms of these diseases. Despite significant advances in understanding the underlying mechanisms of neurodegeneration, there remains a need for methods and compositions for neuroprotection. -1- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0004] Cannabinoids, salts thereof, and derivatives thereof, have several properties with therapeutic potential. Activation or blocking of CB1 and / or CB2 receptors with a cannabinoid can regulate downstream signaling and metabolic pathways and subsequently influence synaptic transmission, including transmission of pain and other sensory signals in the periphery, immune response, and inflammation. Thus, there is an interest in the use of natural or non-natural cannabinoids for therapeutic purposes. SUMMARY

[0005] Described herein are compounds, pharmaceutical compositions, and methods of their use. In one or more embodiments, the compounds and pharmaceutical compositions are neuroprotective. A compound or a pharmaceutical composition containing a compound, for example of Formula I, can be contacted with a neuron, thereby providing a neuroprotective effect. In one or more embodiments, the contacting is performed by administering the compound, for example of Formula I, or pharmaceutical composition comprising such a compound, for example of Formula I, to a subject in need thereof. The compound, for example of Formula I, pharmaceutical compositions comprising such a compound, for example of Formula I, and related methods are useful in providing neuroprotection and in some embodiments treating a variety of ocular neurodegenerative diseases. In one or more embodiments, compounds, for example of Formula I, and pharmaceutical compositions comprising such a compound, for example of Formula I, are provided for use in inducing a neuroprotective effect in, for example, retinal neurons. For example, a compound, for example of Formula I, or pharmaceutical composition including such a compound, for example of Formula I, can be locally or systemically administered to a subject to induce a neuroprotective effect in retinal neurons, e.g., to treat an ocular neurodegenerative disease such as AMD (including, for example, atrophic (non-exudative or “dry”) AMD and neovascular (exudative or “wet”) AMD). In some embodiments, the compound, pharmaceutical compositions protects against or treats neurodegeneration in a neuron, including that which causes or is caused by neuronal atrophy, axonal degeneration (e.g., Wallerian and / or Wallerian-like degeneration), and / or induction of necrotic or programmed mechanisms of cell death (including, for example, programmed cell death such as apoptosis, autophagy, pyroptosis, and oncosis in neurons); and / or protects against or treats neurodegeneration caused by physical injury, oxidative stress, excitotoxicity, mitochondrial dysfunction, inflammation (including neuroinflammation), iron accumulation, and / or protein aggregation. Cannabinoids, including a compound of Formula I are involved in transmission pain and other sensory signals in the periphery, in immune response, and in inflammation. A compound of the first or second aspect -2- 1101228610\3\AMERICASAttorney Docket No.120754.00100 (and any embodiments thereof) or a pharmaceutical composition of the third aspect (and any embodiments thereof) can be used to protect against or treat the conditions in this paragraph.

[0006] In a first aspect, provided is a compound of Formula I: (named 3-(4- [c]chromen-1-ol or 4-chlorobutyl-cannabinol, or, derivative thereof; and / or a pharmaceutically acceptable salt thereof. In one or more embodiments, the compound of Formula I is provided as a derivative thereof. In one or more embodiments, the derivative of the compound of Formula I is a prodrug of Formula I.

[0007] In a second aspect, provided is a compound of Formula I: or a pharmaceutically acceptable embodiments, the compound ofFormula I is not provided as a

[0008] In a third aspect, provided is a pharmaceutical composition comprising a compound of Formula I: or a derivative thereof; and / or athereof, and a pharmaceutically acceptable excipient. In one or more embodiments, provided is a pharmaceutical composition comprising a compound of Formula I, according to: -3- 1101228610\3\AMERICASAttorney Docket No.120754.00100 ;and a pharmaceutically some embodiments, the pharmaceutical composition comprises a compound of Formula I or a derivative thereof; and / or a pharmaceutically acceptable salt thereof; and the pharmaceutically acceptable excipient comprises ethanol. In some embodiments, the pharmaceutical composition comprises a compound of Formula I or a derivative thereof; and / or a pharmaceutically acceptable salt thereof; and the pharmaceutically acceptable excipient does not comprise ethanol. In some embodiments, the pharmaceutically acceptable excipient consists solely of ethanol. In some embodiments, the pharmaceutically acceptable excipient does not consist solely of ethanol.

[0009] In a fourth aspect, provided is a method of protecting a neuron from neurodegeneration or treating neurodegeneration in a neuron, the method comprising contacting the neuron with a compound of the first or second aspect (and any embodiments thereof) or a pharmaceutical composition of the third aspect (and any embodiments thereof), in an amount (or concentration) sufficient to inhibit neurodegeneration to a subject in need thereof. In one or more embodiments, the contacting is in vitro. In one or more embodiments, the contacting is in vivo. In one or more embodiments, the contacting comprises administering the compound or pharmaceutical composition to a subject in need thereof. In one or more embodiments, the neuron is a retinal neuron (e.g., retinal ganglion). In one or more embodiments, the contacting comprises administering the compound of the first or second aspect (and any embodiments thereof) or pharmaceutical composition of the third aspect (and any embodiments thereof), e.g., as an intravitreal (IVT) injection. The intravitreal injection may target the back of the eye as shown in FIG.1 (arrow). In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt. In one or more embodiments, the compound of Formula I is 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]chromen-1-ol (i.e., not a derivative or pharmaceutically acceptable salt thereof). In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing -4- 1101228610\3\AMERICASAttorney Docket No.120754.00100 eye pressure in a subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma.

[0010] In a fifth aspect, provided is a method of protecting a subject from neurodegeneration or treating neurodegeneration in a subject comprising administering a compound of the first or second aspect (and any embodiments thereof) or a pharmaceutical composition of the third aspect (and any embodiments thereof) to a subject in need thereof. In one or more embodiments, the pharmaceutical composition is suitable to achieve a neuroprotective dose of 4-chlorobutyl-CBN, or a pharmaceutically acceptable salt thereof, or a derivative (e.g., prodrug) thereof. In one or more embodiments, the compound of the first or second aspect (and any embodiments thereof) or pharmaceutical composition of the third aspect (and any embodiments thereof) is formulated for administration to the eye and is for use in protecting a neuron from neurodegeneration or treating neurodegeneration in a neuron. In one or more embodiments, the compound of the first or second aspect (and any embodiments thereof) or pharmaceutical composition of the third aspect (and any embodiments thereof) is formulated to achieve a concentration of from about 0.15 µM to less than about 20 µM 4-chlorobutyl-CBN, or a pharmaceutically acceptable salt thereof, or a derivative (e.g., prodrug) thereof, in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the ocular neuronal cell is a retinal neuron or other neuronal cell (e.g., including but not limited to a retinal ganglion and a photoreceptor). In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing eye pressure in a subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma.

[0011] In a sixth aspect, provided is a use of the compound of the first or second aspect (and any embodiments thereof) or pharmaceutical composition of the third aspect (and any embodiments thereof), for treatment of neurodegeneration in a subject in need thereof, preferably according to one or more of the foregoing aspects (in the fourth and fifth aspects), and any embodiments thereof provided herein, with a pharmaceutical composition described herein, or according to a method described herein. In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing eye pressure in a -5- 1101228610\3\AMERICASAttorney Docket No.120754.00100 subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma.

[0012] In a seventh aspect, provided is a method of treating age-related macular degeneration, the method comprising administering a compound of the first or second aspect (and any embodiments thereof) or a pharmaceutical composition of the third aspect (and any embodiments thereof) to a subject in need thereof.

[0013] In an eighth aspect, provided is a method of improving RPE integrity in an AMD subject and reducing autofluorescence deposits – a hallmark of dry AMD, comprising administering a compound of the first or second aspect (and any embodiments thereof) or a pharmaceutical composition of the third aspect (and any embodiments thereof) to a subject in need thereof. In one or more embodiments, provided is a method wherein an amount is administered and is sufficient for improving retinal pigment epithelium integrity.

[0014] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the contacting and / or administering comprises topical administration to a subject in need thereof. For example, the administration is to a subject suffering from neurodegeneration, e.g., neurodegeneration of the eye. In one or more embodiments, the contacting and / or administering comprises administration to a subject suffering from a neurodegenerative disease, e.g., a neurodegenerative disease of the eye. In one or more embodiments, the contacting and / or administering comprises administration to a subject suffering from AMD. In one or more embodiments, the contacting and / or administering comprises administration to a subject diagnosed with AMD. In one or more embodiments, the method comprises simultaneously or sequentially contacting and / or administering to a subject in need thereof an additional active agent for treatment of AMD. In some or any embodiments, the subject is suffering from glaucoma. In some or any embodiments, the subject is not suffering from glaucoma.

[0015] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the amount is an amount (or concentration) sufficient to reduce an amount or rate of apoptosis of a population of neurons contacted with the compound or pharmaceutical composition. In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration is an amount (or concentration) sufficient to reduce the progression of age-related macular degeneration or vision loss associated therewith. In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt. In one or more embodiments, the compound of Formula I is 3-(4-chlorobutyl)-6,6,9- trimethyl-6H-benzo[c]chromen-1-ol (i.e., not a derivative or pharmaceutically acceptable salt -6- 1101228610\3\AMERICASAttorney Docket No.120754.00100 thereof). In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect retinal neurons, protect retinal tissue, protect photoreceptor cells, protect against accumulation of or reduce autofluorescent extracellular debris, protect against or treat neuroinflammation. In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect against or treat neuroinflammation. In some or any embodiments, the subject is suffering from glaucoma. In some or any embodiments, the subject is not suffering from glaucoma.

[0016] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the amount (or concentration) sufficient to inhibit neurodegeneration or the amount (or concentration) sufficient to improve retinal pigment epithelium integrity is from about 0.15 µM to less than about 20 µM of 4-chlorobutyl-CBN (or a pharmaceutically acceptable salt thereof, or a derivative (e.g., a prodrug); in some embodiments, wherein the amount is of 4-chlorobutyl-CBN – not as a salt and not as a prodrug) thereof in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration or the amount (or concentration) sufficient to improve retinal pigment epithelium integrity is an amount that results in a concentration selected from about 0.3 µM to less than or equal to about 15 µM of 4-chlorobutyl-CBN in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration or the amount (or concentration) sufficient to improve retinal pigment epithelium integrity is an amount that results in a concentration of greater than or equal to about 0.5 µM to less than or equal to about 15 µM, such as a concentration selected from about 1 µM to about 15 µM, about 2 µM to about 15 µM, about 3 µM to about 15 µM, about 2 µM to about 12 µM, about 3 µM to less than about 12 µM, about 3 µM to less than about 11 µM, about 3 µM to about 10 µM, or 3 µM to 10 µM of 4-chlorobutyl-CBN in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt. In one or more embodiments, the compound of Formula I is 3-(4-chlorobutyl)-6,6,9-trimethyl- 6H-benzo[c]chromen-1-ol (i.e., not a derivative or pharmaceutically acceptable salt thereof). In one or more embodiments, the amount is based on the compound excluding any pharmaceutically acceptable salt. In some or any embodiments, the subject is suffering from glaucoma. In some or any embodiments, the subject is not suffering from glaucoma.

[0017] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the amount (or concentration) sufficient to inhibit -7- 1101228610\3\AMERICASAttorney Docket No.120754.00100 neurodegeneration or the amount (or concentration) sufficient to improve retinal pigment epithelium integrity is an amount that results in a concentration of selected from about 0.15 µM to less than about 20 µM, such as selected from about 0.5 µM to less than about 15 µM, selected from greater than about 2 µM to less than about 12 µM, from 2 µM to 12 µM, selected from about 3 µM to less than about 12 µM, selected from about 3 µM to less than about 11 µM, selected from about 3 µM to about 10 µM, or selected from 3 µM to 10 µM of 4-chlorobutyl-CBN, a pharmaceutically acceptable salt thereof, or a derivative (e.g., a prodrug) thereof in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the subject is suffering from AMD. In some or any embodiments, the subject is suffering from glaucoma. In some or any embodiments, the subject is not suffering from glaucoma.

[0018] In one or more embodiments, the amount (or concentration) sufficient to inhibit neurodegeneration or the amount (or concentration) sufficient to improve retinal pigment epithelium integrity is an amount that results in a concentration range with a lower end of the range selected from any one of 0.15, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7 ,8, and 9 µM, and with an upper end of the range selected from any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 µM, where the lower end and the upper end form a mathematically allowed range, and the concentration is of 4-chlorobutyl-CBN, a pharmaceutically acceptable salt thereof, or a derivative (e.g., a prodrug) thereof in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the subject is suffering from AMD. In some or any embodiments, the subject is suffering from glaucoma. In some or any embodiments, the subject is not suffering from glaucoma.

[0019] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the pharmaceutical composition has a final concentration of the compound of Formula I (in vivo) that is about 1 / 10 (one tenth) of an initial concentration of the compound of Formula I. In one or more embodiments, the pharmaceutical composition has a final concentration of the compound of Formula I (in vivo) that is 1 / 10 of an initial concentration of the compound of Formula I. In one or more embodiments, the initial concentration is the concentration of 4-chlorobutyl-CBN (or a pharmaceutically acceptable salt thereof, or a derivative thereof) in the pharmaceutical composition. In one or more embodiments, wherein when the pharmaceutical composition is an intravitreal injection, the initial concentration is the concentration of 4-chlorobutyl-CBN (or a pharmaceutically acceptable salt thereof, or a derivative thereof) in the intravitreal injection. In one or more embodiments, the final concentration is the concentration of 4-chlorobutyl-CBN (or a pharmaceutically acceptable salt thereof, or a derivative -8- 1101228610\3\AMERICASAttorney Docket No.120754.00100 thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the final concentration is the concentration of 4-chlorobutyl-CBN (or a pharmaceutically acceptable salt thereof, or a derivative thereof) in the vitreous humor. In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt. In one or more embodiments, the compound of Formula I is 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]chromen-1-ol (i.e., not a derivative or pharmaceutically acceptable salt thereof). In one or more embodiments, the concentration is based on the compound excluding any pharmaceutically acceptable salt.

[0020] In one or more embodiments, the 4-chlorobutyl-CBN is provided in an extended-release formulation. In one or more embodiments, the formulation comprises: a) a delivery carrier comprising a cellulosic polymer and an anionic polysaccharide; and b) nanoparticles comprising an amphiphilic non-ionizable block copolymer and 4-chlorobutyl-CBN, wherein the formulation has a gel point of about 30°C to about 37°C.

[0021] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the contacting and / or administering comprises systemic administration. In one or more embodiments, the systemic administration comprises intravenous injection. In one or more embodiments, the systemic administration comprises oral administration. In one or more embodiments, the systemic administration comprises transdermal administration. In one or more embodiments, the systemic administration comprises intravitreal injection. In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt. In one or more embodiments, the compound of Formula I is 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]chromen-1-ol (i.e., not a derivative or pharmaceutically acceptable salt thereof). In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing eye pressure in a subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma.

[0022] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth aspects (and any embodiments of the aspects), the contacting and / or administration comprises local administration. In one or more embodiments, the contacting and / or administering is directly to the eye. For example, the contacting and / or administering is onto the eye (e.g., as an eyedrop such as in the form of a microemulsion eyedrop, or an eye gel). As another example, the contacting and / or -9- 1101228610\3\AMERICASAttorney Docket No.120754.00100 administering is directly into the eye, or to the back of the eye (e.g., via intravitreal injection or pump). In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing eye pressure in a subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma. INCORPORATION BY REFERENCE

[0023] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE FIGURES

[0024] FIG.1 illustrates a half section view of an eye, indicating parts of the eye and showing the site of intravitreal injection to treat age-related macular degeneration according to one or more embodiments herein.

[0025] FIG. 2A illustrates flash electroretinogram (fERG) test results, showing the response of photoreceptors in the eye to light stimuli at an intensity (luminance) of 0.01 cd*s / m^2. The a-wave amplitude (µV) indicates the response of the photoreceptors, and the luminance (cd*s / m^2) indicates the intensity of the flash of light. Control: Healthy animals with no light damage (LD) and no treatment. Vehicle: light damage and treatment with sterile Dilution Buffer.

[0026] FIG. 2B illustrates flash electroretinogram (fERG) test results, showing the response of photoreceptors in the eye to light stimuli at an intensity (luminance) of 0.01 cd*s / m^2. The % a- wave amplitude normalized to healthy animals (CTRL or control = 100%) is shown. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with Dilution Buffer.

[0027] FIG. 3A illustrates flash electroretinogram (fERG) test results, showing the response of photoreceptors in the eye to light stimuli at increasing levels of intensity. The a-wave amplitude (µV) indicates the response of the photoreceptors, and the luminance (cd*s / m^2) indicates the intensity of the flash of light. For eyes injected with 4-chlorobutyl-CBN, an increase of a-wave amplitude was observed compared to the vehicle, for the respective light stimulus (measured in luminance). For eyes injected with CBN, no overall improvement of a-wave amplitude was -10- 1101228610\3\AMERICASAttorney Docket No.120754.00100 observed compared to the vehicle. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0028] FIG.3B illustrates the a-wave amplitude at an intensity of 0.01 cd*s / m^2 (from FIG.3A), to better represent the increase of a-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the a-wave amplitude in µV (left) and the % a-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0029] FIG.3C illustrates the a-wave amplitude at an intensity of 0.1 cd*s / m^2 (from FIG.3A), to better represent the increase of a-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the a-wave amplitude in µV (left) and the % a-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0030] FIG.3D illustrates the a-wave amplitude at an intensity of 1 cd*s / m^2 (from FIG.3A), to better represent the increase of a-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the a-wave amplitude in µV (left) and the % a-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0031] FIG. 3E illustrates the a-wave amplitude at an intensity of 10 cd*s / m^2 (from FIG.3A), to better represent the increase of a-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the a-wave amplitude in µV (left) and the % a-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0032] FIG.3F illustrates the a-wave amplitude at an intensity of 100 cd*s / m^2 (from FIG.3A), to better represent the increase of a-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the a-wave amplitude in µV (left) and the % a-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0033] FIG. 4A illustrates flash electroretinogram (fERG) test results, showing the response of the inner retina in the eye to light stimuli at increasing levels of intensity. The b-wave amplitude (µV) indicates the response of the inner retina, and the luminance (cd*s / m^2) indicates the intensity of the flash of light. For eyes injected with 4-chlorobutyl-CBN, an increase of b-wave amplitude was observed compared to the vehicle, for the respective light stimulus (measured in luminance). For eyes injected with CBN, no overall improvement of b-wave amplitude was -11- 1101228610\3\AMERICASAttorney Docket No.120754.00100 observed compared to the vehicle. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0034] FIG.4B illustrates the b-wave amplitude at an intensity of 0.1 cd*s / m^2 (from FIG.4A), to better represent the increase of b-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the b-wave amplitude in µV (left) and the % b-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0035] FIG.4C illustrates the b-wave amplitude at an intensity of 1 cd*s / m^2 (from FIG.4A), to better represent the increase of b-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the b-wave amplitude in µV (left) and the % b-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0036] FIG. 4D illustrates the b-wave amplitude at an intensity of 10 cd*s / m^2 (from FIG.4A), to better represent the increase of b-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the b-wave amplitude in µV (left) and the % b-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0037] FIG.4E illustrates the b-wave amplitude at an intensity of 100 cd*s / m^2 (from FIG.4A), to better represent the increase of b-wave amplitude that occurs upon 4-chlorobutyl-CBN treatment. The histograms show the b-wave amplitude in µV (left) and the % b-wave amplitude normalized to healthy animals (right) (CTRL or control = 100%). Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0038] FIG.5 illustrates flash electroretinogram (fERG) test results, showing the response of the inner retinal circuitry in the eye to light stimuli (mainly due to rod bipolar and A11 / A17 amacrine cells) at increasing levels of intensity. The oscillatory potentials (OPs) amplitude (µV) indicates the response of the inner retina, and the luminance (cd*s / m^2) indicates the intensity of the flash of light. For eyes injected with 4-chlorobutyl-CBN, an increase of OPs amplitude was observed compared to the vehicle for the respective light stimulus (measured in luminance). For eyes injected with CBN (30 µM), an increase of the OPs amplitude at 100 cd*s / m^2 (scotopic stimulus) was observed compared to the vehicle, whereas no overall improvement of OPs amplitude was observed compared to the vehicle for other light stimulus. For eyes injected with CBN (100 µM), no overall improvement of OPs amplitude was observed compared to the vehicle, for any light stimulus applied. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer. -12- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0039] FIG.6A illustrates in vivo imaging of the inner retinal layer structure of the light damaged rat model. The imaging shows the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL) in cells treated with 4-chlorobutyl-CBN (3 µM), Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0040] FIG. 6B illustrates, an inverse (negative) exposure, in vivo imaging of the inner retinal layer structure. The imaging shows the outer nuclear layer, inner nuclear layer, and ganglion cell layer in cells treated with 4-chlorobutyl-CBN (3 µM), Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0041] FIG. 7A illustrates a-wave amplitudes in response to different dose levels of 4-Cl-Butyl- CBN 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). An overall increase of the a-wave response is observed in the eye treated with 4-Cl-Butyl-CBN 3 µM and 10 µM compared to the vehicle. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0042] FIG. 7B illustrates b-wave amplitudes in response to different dose levels of 4-Cl-Butyl- CBN 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). An overall increase of the b-wave response is observed in the eye treated with 4-Cl-Butyl-CBN 3 µM and 10 µM compared to the vehicle. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0043] FIG. 7C illustrates oscillatory potentials (OPs) amplitudes in response to different dose levels of 4-Cl-Butyl-CBN 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). An overall increase of the OPs response is observed in the eye treated with 4-Cl-Butyl-CBN 3 µM and 10 µM compared to the vehicle. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer.

[0044] FIG.8A illustrates histological imaging of the retina to measure outer nuclear layer (ONL) thickness. Scale bar: 200 µm.

[0045] FIG.8B illustrates the ratio of ONL: retina thickness at various positions along the superior retina and the inferior retina. Data are shown as mean + / - SE. Statistical analysis was Student’s t- test between vehicle and 4-Cl-Butyl -CBN (3 µM) (n=7 / group). *p<0.05; ***p<0.001 versus vehicle. An increase in ONL: retina thickness is observed for 4-Cl-Butyl-CBN compared to the vehicle.

[0046] FIG.8C illustrates a schematic representation of a retinal cryosection showing the retinal locations indicated in FIG.8B along the superior retina and inferior retina. ON=optic nerve. -13- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0047] FIG.9A illustrates histological imaging of autofluorescent (AF) extracellular deposits. A decrease in extracellular deposits is observed for 4-Cl-Butyl-CBN compared to the vehicle.

[0048] FIG. 9B illustrates the number of AF deposits. A statistically significant decrease in extracellular deposits is observed for 4-Cl-Butyl-CBN compared to the vehicle. Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl-Butyl-CBN (3 µM) (n=7 / group). **p<0.01 versus vehicle.

[0049] FIG. 10A illustrates histological imaging of retinal pigment epithelium (RPE) with anti- RPE65. An improvement in RPE integrity is observed for 4-Cl-Butyl-CBN compared to the vehicle.

[0050] FIG. 10B illustrates the RPE length normalized to the Control. A statistically significant increase in RPE length was observed between 4-Cl-Butyl-CBN compared to the Control. Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl-Butyl- CBN (3 µM) (n=7 / group). ***p<0.001 versus vehicle. These data show that 4-Cl-Butyl-CBN improved RPE length in an AMD model.

[0051] FIG.11A illustrates immunostaining of retinal cross-sections for IBA-1. Scale bar: 50 µm. A reduction in inflammation in the retina is observed for 4-Cl-Butyl-CBN compared to the vehicle.

[0052] FIG.11B illustrates the number of IBA-1 (+) cells counter in the outer retina (subretinal, outer nuclear layer, outer plexiform layer) from the superior to the inferior edge of the retina. A reduction in inflammation in the retina is observed for 4-Cl-Butyl-CBN compared to vehicle Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl- Butyl-CBN (3 µM) (n=7 / group). *p<0.05; **p<0.01 versus vehicle.

[0053] FIG. 11C illustrates the number of IBA-1 (+) cells in the superior retina. A statistically significant reduction in inflammation in the superior retina is observed for 4-Cl-Butyl-CBN compared to vehicle. Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl-Butyl-CBN (3 µM) (n=7 / group). *p<0.05; **p<0.01 versus vehicle.

[0054] FIG. 11D illustrates the number of IBA-1 (+) cells in the inferior retina. A statistically significant reduction in inflammation in the inferior retina is observed for 4-Cl-Butyl-CBN compared to vehicle. Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl-Butyl-CBN (3 µM) (n=7 / group). *p<0.05; **p<0.01 versus vehicle.

[0055] FIG. 11E illustrates the number of IBA-1 (+) cells in the superior and inferior retina. A statistically significant reduction in inflammation in the retina is observed for 4-Cl-Butyl-CBN compared to vehicle. Data are shown as mean + / - SE. Statistical analysis was Student’s t-test between vehicle and 4-Cl-Butyl-CBN (3 µM) (n=7 / group). *p<0.05; **p<0.01 versus vehicle. -14- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0056] FIG.12 illustrates the concentration of 4-Cl-Butyl-CBN as applied topically to a rabbit or IVT to a rat. An increased concentration is observed in retina / RPE / choroid for IVT administration in rat versus topical application in rabbit. DETAILED DESCRIPTION

[0057] Described herein are compounds, for example of Formula I, and pharmaceutical compositions thereof and methods of such compounds, for example of Formula I, and pharmaceutical compositions for protecting neurons from one or more cytotoxic stimuli. In one or more embodiments, the method includes contacting the neurons with the compound, for example of Formula I, or pharmaceutical composition thereof, such as by administering the compound or pharmaceutical composition to a subject in need thereof. The methods, compounds, for example of Formula I, and pharmaceutical compositions thereof described herein find particular use, but are not limited to, protection of retinal neurons. In some cases, the methods, compounds, for example of Formula I, and pharmaceutical compositions thereof described herein can be used for neuroprotection of retinal neurons in a subject, such as a subject suffering from age-related macular degeneration. In this manner, the methods, compounds, for example of Formula I, and pharmaceutical compositions thereof may be described as neuroprotective. In one or more embodiments, the neuron is damaged, including but not limited, to a light damaged neuron. In one or more embodiments, the compound, for example of Formula I, and pharmaceutical composition thereof includes 4-chlorobutyl-CBN. In some cases, the method includes contacting retinal neurons with 4-chlorobutyl-CBN, such as by administering 4-chlorobutyl-CBN (or a pharmaceutical composition thereof) to a subject in need thereof. In one or more embodiments, the method is for protecting a subject from AMD or for treating AMD in a subject in need thereof. In some or any embodiments, the method is for protecting a subject from glaucoma or for treating glaucoma in a subject. In some or any embodiments, the method is not for protecting a subject from glaucoma and not for treating glaucoma in a subject. In some or any embodiments, the method is for reducing eye pressure in a subject with glaucoma. In some or any embodiments, the method is not for reducing eye pressure in a subject with glaucoma.

[0058] In one or more embodiments, the compound is 4-chlorobutyl-CBN having formula of Formula I:-15- 1101228610\3\AMERICASAttorney Docket No.120754.00100 I. In one or more embodiments, the compound is a compound of Formula I, or a pharmaceutically acceptable salt thereof.

[0059] The pharmaceutical composition can be or can contain 4-chlorobutyl-CBN having the formula of Formula I:

[0060] In one or more comprises a compound of Formula I or a pharmaceutically acceptable excipient. In one or more embodiments, the pharmaceutical composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0061] In one or more embodiments, the compound or pharmaceutical composition is or can contain a prodrug of the compound of Formula I or pharmaceutically acceptable salts. For example, the compound or pharmaceutical composition is or can contain a prodrug of 4- chlorobutyl-CBN, or a pharmaceutically acceptable salt thereof.

[0062] In some cases, the prodrug is an ester of 4-chlorobutyl-CBN (according to the formula of Formula I) or a pharmaceutically acceptable salt thereof. In some cases, the prodrug is a D-(-)-glyceric acid ester of 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof. Additional prodrug strategies for the compounds described herein can be found in U.S. Pat. Publ. Nos. 2016 / 0228490; 2011 / 0052694; 2015 / 0197484; 2008 / 0076789; 2009 / 0143462; 2012 / 0289484; 2009 / 0036523; 2009 / 0156814; and 2008 / 0008745; and Adelli et al., Investigative Ophthalmology & Visual Science, April 2017, Vol.58, No.4, p.2168; and Upadhye et al., AAPS PharmSciTech, Vol. 11, No.2, June 2010, p.509, the contents of which are hereby incorporated in the entirety for all purposes and in particular for the 4-chlorobutyl-CBN prodrugs and pharmaceutical compositions comprising them, and methods of using and / or administering such prodrugs described therein.

[0063] As used herein, the term “prodrug” refers to a derivative that is a precursor compound that, following administration, releases the biologically active compound in vivo via some chemical or physiological process (e.g., a prodrug on reaching physiological pH or through enzyme action is converted to the biologically active compound). A prodrug itself may either lack or possess the -16- 1101228610\3\AMERICASAttorney Docket No.120754.00100 desired biological activity. Thus, the term “prodrug” refers to a precursor of a biologically active compound that is pharmaceutically acceptable. In some cases, a prodrug has improved physical and / or delivery properties over a parent compound from which the prodrug has been derived. The prodrug often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (H. Bundgard, Design of Prodrugs (Elsevier, Amsterdam, 1988), pp. 7-9, 21-24). A discussion of prodrugs is provided in T. Higuchi et al., “Pro-Drugs as Novel Delivery Systems,” ACS Symposium Series, Vol.14 and in E.B. Roche, ed., Bioreversible Carriers in Drug Design (American Pharmaceutical Association & Pergamon Press, 1987). Exemplary advantages of a prodrug can include, but are not limited to, its physical properties, such as enhanced drug stability for long-term storage.

[0064] The term “prodrug” is also meant to include any covalently bonded carriers which release the active compound in vivo when the prodrug is administered to a subject. Prodrugs of a therapeutically active compound, as described herein, can be prepared by modifying one or more functional groups present in the therapeutically active compound, i.e., the compound of Formula I, or a pharmaceutically acceptable salt thereof, and other therapeutically active compounds used in methods according to one or more embodiments or included in the pharmaceutical compositions according to one or more embodiments, in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to yield the parent therapeutically active compound. Prodrugs include compounds of Formula I wherein the hydroxy is covalently bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy group. Examples of prodrugs include, but are not limited to, formate or benzoate derivatives of an alcohol available for reaction, and the like.

[0065] In some cases, the prodrug comprises a 3,6,9,12-tetraoxatridecanoyl ester; an N,N-dimethylglycyl ester; a 3,6,9,12-tetraoxatridecyl carbonate; an N-formylglycyl ester; an N-formylsarcosyl ester; a 3,6,9,12-tetraoxatridecyl oxalate; a hemisuccinate; a 4-aminobutyl carbamate; a prolyl ester; a 3-dimethylamino propionate; a glycolate; a (D)-ribonate; a phosphate ammonium salt; an (R)-2,3-dihydroxypropyl carbonate; a 3-hydroxy-2-(hydroxymethyl)-2- methylpropanoate; a glycinate; a β-alaninate; an (S)-2,3-dihydroxypropanoate; an (S)-2,3-dihydroxypropyl carbonate; or an (R)-2,3-dihydroxypropyl carbonate.

[0066] Similarly, if a disclosed compound or a pharmaceutically acceptable form of the compound contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as (C1-C6)alkanoyloxymethyl, 1-((C1-C6))alkanoyloxy)ethyl, 1-methyl-1-((C1-C6)alkanoyloxy)ethyl, (C1-C6)alkoxycarbonyloxymethyl, N(C1-C6)alkoxycarbonylaminomethyl, succinoyl, -17- 1101228610\3\AMERICASAttorney Docket No.120754.00100 (C1-C6)alkanoyl, ^-amino(C1-C4)alkanoyl, arylacyl and ^-aminoacyl, or ^-aminoacyl- ^- aminoacyl, where each ^-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH)2, P(O)(O(C1-C6)alkyl)2 or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate).

[0067] The use of prodrug systems is described in T. Järvinen et al., “Design and Pharmaceutical Applications of Prodrugs” in Drug Discovery Handbook (S.C. Gad, ed., Wiley-Interscience, Hoboken, NJ, 2005), ch.17, pp.733-796. Other alternatives for prodrug construction and use are known in the art. When a method or pharmaceutical composition according to one or more embodiments uses or includes a prodrug of cannabinol or 4-chlorobutyl-CBN, or other therapeutically active agent, prodrugs and active metabolites of a compound may be identified using techniques known in the art. See, e.g., Bertolini et al., J. Med. Chem., 40, 2011-2016 (1997); Shan et al., J. Pharm. Sci., 86 (7), 765-767; Bagshawe, Drug Dev. Res., 34, 220-230 (1995); Bodor, Advances in Drug Res., 13, 224-331 (1984); Bundgaard, Design of Prodrugs (Elsevier Press 1985); Larsen, Design and Application of Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al., eds., Harwood Academic Publishers, 1991); Dear et al., J. Chromatogr. B, 748, 281-293 (2000); Spraul et al., J. Pharmaceutical & Biomedical Analysis, 10, 601-605 (1992); and Prox et al., Xenobiol., 3, 103-112 (1992).

[0068] Exemplary prodrugs useful in one or more embodiments of the present disclosure include but are not limited to the following prodrugs of the compound of Formula I or a pharmaceutically acceptable salt thereof: ;

[0069] wherein X and Y, can beare selected from the group consisting of: hydrogen, alkali metals (e.g., sodium and potassium), alkaline earth metals (e.g., calcium and magnesium); and cations of pharmaceutically acceptable organic amines (e.g., quaternated or protonated amines, including alkylamines, hydroxyalkylamines, monoamines, diamines, and naturally occurring amines). Examples of such pharmaceutically acceptable organic bases include choline, betaine, caffeine, N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, hydrabamine, isopropylamine, methylglucamine, morpholine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, tetramethylammonium hydroxide, benzyltrimethylammonium hydroxide, -18- 1101228610\3\AMERICASAttorney Docket No.120754.00100 tris(hydroxymethyl)aminomethane (TRIS), N-(2-hydroxyethyl)pyrrolidine, piperazine, glucosamine, arginine, lysine and histidine. In a one or more embodiments, X and Y are different substituent groups. In one or more embodiments, X and Y are the same substituent group. In one or more embodiments, X and Y can both be part of the same functional group, such as piperazine. In one or more embodiments, the phosphate is selected from a group consisting of a diphosphate and triphosphate.

[0070] In one or more embodiments, the prodrug is a prodrug of the compound of Formula I or a pharmaceutically acceptable salt thereof; ; wherein R4is alkyl, alkoxyalkyl,or hydroxyalkylamino; in one or more embodiments, R4comprises from 1 to 12 carbons and optionally no more than 4 substitutions; in one or more embodiments, R4comprises from 1 to 6 carbons and optionally no more than 2 substitutions.

[0071] In one or more embodiments, the prodrug is a prodrug of the compound of Formula I or a pharmaceutically acceptable salt thereof; ; wherein R4is alkyl, alkoxyalkyl,hydroxyalkylamino; in one or more embodiments, R4comprises from 1 to 12 carbons and optionally no more than 4 substitutions; in one or more embodiments, R4comprises from 1 to 6 carbons and optionally no more than 2 substitutions.

[0072] In one or more embodiments, the prodrug is a prodrug of the compound of Formula I or a pharmaceutically acceptable salt thereof; ; wherein R4is alkyl, alkoxyalkyl,hydroxyalkylamino; in one or more embodiments, R4comprises from 1 to 12 carbons and optionally no more than 4 substitutions; in -19- 1101228610\3\AMERICASAttorney Docket No.120754.00100 one or more embodiments, R4comprises from 1 to 6 carbons and optionally no more than 2 substitutions.

[0073] In one or more embodiments, the prodrug is a prodrug of the compound of Formula I or a pharmaceutically acceptable salt thereof; wherein R4is alkyl, alkoxyalkyl, hydroxyalkylamino; in one or more 4embodiments, R comprises no more than 4 substitutions; in one or more embodiments, R4comprises from 1 to 6 carbons and optionally no more than 2 substitutions.

[0074] Prodrugs useful in one or more embodiments of the present disclosure include but are not limited to the following prodrug of the compound of Formula I or a pharmaceutically acceptable salt thereof:

[0075] Prodrugs useful in one ordisclosure include but are not limited to the following prodrug the compound of Formula I or a pharmaceutically acceptable salt thereof: O Cl .

[0076] Prodrugs useful in one orpresent disclosure include but are not limited to the following prodrugs of the compound of Formula I or a pharmaceutically acceptable salt thereof: -20- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0077] In one or formulatedwith a as hydroxypropyl beta- cyclodextrin, or sulfobutyl ether beta-cyclodextrin.

[0078] The pharmaceutical composition can contain additional active agents. In one or more embodiments, the pharmaceutical composition can contain 4-chlorobutyl-CBN, a pharmaceutically acceptable salt thereof, or a derivative thereof, and an additional cannabinoid or a terpenoid. In one or more embodiments, the pharmaceutical composition can contain an additional active pharmaceutical agent for treatment of age-related macular degeneration.

[0079] Currently, different classes of therapeutic agents are used for the treatment of age-related macular degeneration, including but not limited to: micronutrient supplementation, vascular- endothelial derived growth factor (VEGF)-A antagonists (anti-VEGF agents or VEGF inhibitors), photodynamic therapy.

[0080] Useful micronutrient supplements include but are not limited to vitamin C, zinc, vitamin E, copper, and beta carotene. Useful anti-VEGF agents include but are not limited to ranibizumab, aflibercept, brolucizumab, faricimab, and bevacizumab. A useful photodynamic therapy agent includes but is not limited to verteporfin. In one or more embodiments, different therapeutic agents may be combined.

[0081] In one or more embodiments, the compounds and pharmaceutical compositions described herein, e.g., containing 4-chlorobutyl-CBN or a pharmaceutically acceptable salt, or derivative thereof, allow a lower dose, or less frequent dosing, of one or more therapeutic agents for treatment of age-related macular degeneration. Definitions

[0082] When referring to the compounds provided herein, the following terms have the following meanings unless indicated otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. In the event that there is a plurality of definitions for a term herein, those in this section prevail -21- 1101228610\3\AMERICASAttorney Docket No.120754.00100 unless stated otherwise. Unless specified otherwise, where a term is defined as being substituted, the groups in the list of substituents are themselves unsubstituted.

[0083] Reference to “about” a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se. For example, description referring to “about X” includes description of “X”. As used herein, and unless otherwise specified, the terms “about” and “approximately,” when used in connection with temperatures, doses, amounts, or weight percent of ingredients of a composition or a dosage form, mean a dose, amount, or weight percent that is recognized by those of ordinary skill in the art to provide a pharmacological effect equivalent to that obtained from the specified dose, amount, or weight percent. Specifically, the terms “about” and “approximately,” when used in this context, contemplate a dose, amount, or weight percent within 15%, within 10%, within 5%, within 4%, within 3%, within 2%, within 1%, or within 0.5% of the specified dose, amount, or weight percent.

[0084] The terms “a” or “an,” as used in herein means one or more, unless context clearly dictates otherwise. For example, “a pharmaceutically acceptable excipient” refers to “one or more pharmaceutically acceptable excipients.”

[0085] As used herein, “alkyl,” refers to a linear or branched saturated monovalent hydrocarbon radical of one to six carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl, butyl, pentyl, and the like. In one or more embodiments, alkyl is (C1-C6)alkyl.

[0086] As used herein, “alkoxy,” refers to an -OR group where R is alkyl, as defined here. In one or more embodiments, alkoxy is (C1-C6)alkoxy.

[0087] As used herein, “(C1-C6)alkanoyloxymethyl,” refers to -CH2OC(O)R where R is (C1-C6)alkyl, as defined herein.

[0088] As used herein, “1-((C1-C6))alkanoyloxy)ethyl,” refers to where R is (C1-C6)alkyl, as defined herein.

[0089] As used herein, “1-methyl-1-((C1-C6)alkanoyloxy)ethyl,” is (C1-C6)alkyl, as defined herein.

[0090] As used herein, “(C1-C6)alkoxycarbonyloxymethyl,” refers to -CH2OC(O)R where R is (C1-C6)alkoxy, as defined herein.

[0091] As used herein, “N(C1-C6)alkoxycarbonylaminomethyl,” refers to -CH2NHC(O)R where R is (C1-C6)alkoxy, as defined herein. -22- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0092] As used herein, “succinoyl,” refers to -C(O)CH2CH2C(O)OH.

[0093] As used herein, “(C1-C6)alkanoyl,” refers to -C(O)R where R is (C1-C6)alkyl, as defined herein.

[0094] As used herein, “ ^-amino(C1-C4)alkanoyl,” refers to -C(O)R where R is alkyl substituted with -NH2.

[0095] As used herein, “arylacyl,” refers to -C(O)phenyl or -C(O)naphthyl.

[0096] As used herein, “alkoxyalkyl,” refers to alkyl, as defined herein, substituted with one or two alkoxy groups, as defined here.

[0097] As used herein, “akylamino,” refers to alkyl, as defined herein, substituted with one or two -NH2.

[0098] As used herein, “hydroxyalkyl,” refers to alkyl, as defined herein, substituted with one or two hydroxy.

[0099] As used herein, “hydroxyalkylamino,” refers to -NHR where R is hydroxyalkyl, as defined herein.

[0100] As used herein, “a subject in need thereof,” and the like, refers to a mammal, preferably a human.

[0101] As used herein, “4-chlorobutyl-cannabinol,” “4-chlorobutyl-CBN, 4-Cl-Butyl-CBN,” or “4-Cl-CBN” refer to 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]chromen-1-ol.

[0102] “Salt” refers to acid or base salts of the compounds used in one or more methods of the present disclosure. Illustrative examples of pharmaceutically acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, and the like) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid and the like) salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like) salts. It is understood that the pharmaceutically acceptable salts are non-toxic. Additional information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

[0103] Thus, when a therapeutically active agent alone or as included in a pharmaceutical composition according to the present disclosure, such as, but not limited to, 4-chlorobutyl-CBN or derivative thereof, possesses a sufficiently acidic, a sufficiently basic, or both a sufficiently acidic and a sufficiently basic functional group, this group or groups can accordingly react with any of a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt of 4-chlorobutyl-CBN. Examples of pharmaceutically acceptable salts include those salts prepared by reaction of the pharmacologically active compound with a mineral or organic acid or an inorganic base, such as salts including sulfates, pyrosulfates, -23- 1101228610\3\AMERICASAttorney Docket No.120754.00100 bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, ^-hydroxybutyrates, glycolates, tartrates, methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates. If the pharmacologically active compound has one or more basic functional groups, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, or with a pyranosidyl acid, such as glucuronic acid or galacturonic acid, or with an alpha-hydroxy acid, such as citric acid, tartaric acid, or with an amino acid, such as aspartic acid, glutamic acid, or with an aromatic acid, such as benzoic acid, cinnamic acid, or with a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like. If the pharmacologically active compound has one or more acidic functional groups, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

[0104] By “pharmaceutically acceptable” it is meant the salt, carrier, diluent, or excipient are compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0105] “Pharmaceutically acceptable excipient” refers to a substance that aids the administration of an active agent to the subject and / or absorption by a subject. Pharmaceutical excipients useful in the present disclosure include, but are not limited to buffers, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors and colors. One of skill in the art will recognize that other pharmaceutical excipients are useful in the present disclosure. -24- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0106] In some cases, protecting groups can be included in compounds used in methods according to the present disclosure or in compounds according to the present disclosure. The use of such a protecting group may be used to prevent subsequent hydrolysis or other reactions that can occur in vivo and can degrade the compound. Groups that can be protected include alcohols, amines, carbonyls, carboxylic acids, phosphates, and terminal alkynes. Protecting groups useful for protecting alcohols include, but are not limited to, acetyl, benzoyl, benzyl, ^-methoxyethoxyethyl ether, dimethoxytrityl, methoxymethyl ether, methoxytrityl, p-methoxybenzyl ether, methylthiomethyl ether, pivaloyl, tetrahydropyranyl, tetrahydrofuran, trityl, silyl ether, methyl ether, and ethoxyethyl ether. Protecting groups useful for protecting amines include carbobenzyloxy, p-methoxybenzylcarbonyl, t-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, acetyl, benzoyl, benzyl, carbamate, p-methoxybenzyl, 3,4-dimethoxybenzyl, p-methoxyphenyl, tosyl, trichloroethyl chloroformate, and sulfonamide. Protecting groups useful for protecting carbonyls include acetals, ketals, acylals, and dithianes. Protecting groups useful for protecting carboxylic acids include methyl esters, benzyl esters, t-butyl esters, esters of 2,6-disubstituted phenols, silyl esters, orthoesters, and oxazoline. Protecting groups useful for protecting phosphate groups include 2-cyanoethyl and methyl. Protecting groups useful for protecting terminal alkynes include propargyl alcohols and silyl groups. Other protecting groups are known in the art.

[0107] As used herein, the term “prodrug” refers to a derivative that is a precursor compound that, following administration, releases the biologically active compound in vivo via some chemical or physiological process (e.g., a prodrug on reaching physiological pH or through enzyme action is converted to the biologically active compound). A prodrug itself may either lack or possess the desired biological activity. Thus, the term “prodrug” refers to a precursor of a biologically active compound that is pharmaceutically acceptable. In certain cases, a prodrug has improved physical and / or delivery properties over a parent compound from which the prodrug has been derived. The prodrug often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (H. Bundgard, Design of Prodrugs (Elsevier, Amsterdam, 1988), pp. 7-9, 21-24). A discussion of prodrugs is provided in T. Higuchi et al., “Pro-Drugs as Novel Delivery Systems,” ACS Symposium Series, Vol.14 and in E.B. Roche, ed., Bioreversible Carriers in Drug Design (American Pharmaceutical Association & Pergamon Press, 1987). Exemplary advantages of a prodrug can include, but are not limited to, its physical properties, such as enhanced drug stability for long-term storage.

[0108] The term “prodrug” is also meant to include any covalently bonded carriers which release the active compound in vivo when the prodrug is administered to a subject. Prodrugs of a -25- 1101228610\3\AMERICASAttorney Docket No.120754.00100 therapeutically active compound, as described herein, can be prepared by modifying one or more functional groups present in the therapeutically active compound, including 4-chlorobutyl-CBN, or a 4-chlorobutyl-CBN derivative, and other therapeutically active compounds used in methods according to the present disclosure or included in pharmaceutical compositions according to the present disclosure, in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to yield the parent therapeutically active compound. Prodrugs include compounds wherein a hydroxy, amino, or mercapto group is covalently bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino, or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, formate or benzoate derivatives of an alcohol or acetamide, formamide or benzamide derivatives of a therapeutically active agent possessing an amine functional group available for reaction, and the like. In some cases, the prodrug is a protecting group modified derivative of the compound, such as a protecting group modified 4-chlorobutyl-CBN or a protecting group modified derivative of 4-chlorobutyl-CBN.

[0109] Similarly, if a disclosed compound or a pharmaceutically acceptable form of the compound contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as (C1-C6)alkanoyloxymethyl, 1-((C1-C6))alkanoyloxy)ethyl, 1-methyl-1-((C1-C6)alkanoyloxy)ethyl, (C1-C6)alkoxycarbonyloxymethyl, N(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (C1-C6)alkanoyl, ^-amino(C1-C4)alkanoyl, arylacyl and ^-aminoacyl, or ^-aminoacyl- ^- aminoacyl, where each ^-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH)2, P(O)(O(C1-C6)alkyl)2 or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate).

[0110] The use of prodrug systems is described in T. Järvinen et al., “Design and Pharmaceutical Applications of Prodrugs” in Drug Discovery Handbook (S.C. Gad, ed., Wiley- Interscience, Hoboken, NJ, 2005), ch.17, pp.733-796. Other alternatives for prodrug construction and use are known in the art. When a method or pharmaceutical composition according to the present disclosure, uses or includes a prodrug of 4-chlorobutyl-CBN, or other therapeutically active agent, prodrugs and active metabolites of a compound may be identified using routine techniques known in the art. See, e.g., Bertolini et al., J. Med. Chem., 40, 2011-2016 (1997); Shan et al., J. Pharm. Sci., 86 (7), 765-767; Bagshawe, Drug Dev. Res., 34, 220-230 (1995); Bodor, Advances in Drug Res., 13, 224-331 (1984); Bundgaard, Design of Prodrugs (Elsevier Press 1985); Larsen, Design and Application of Prodrugs, Drug Design and Development (Krogsgaard- Larsen et al., eds., Harwood Academic Publishers, 1991); Dear et al., J. Chromatogr. B, 748, 281- -26- 1101228610\3\AMERICASAttorney Docket No.120754.00100 293 (2000); Spraul et al., J. Pharmaceutical & Biomedical Analysis, 10, 601-605 (1992); and Prox et al., Xenobiol., 3, 103-112 (1992).

[0111] As used herein, the terms “therapeutically effective quantity,” “therapeutically effective dose,” or “therapeutically effective amount” refer to a dose of one or more compounds or to a dose of pharmaceutical compositions described herein that produces therapeutic effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins). Cannabinoids

[0112] Cannabinoids are a group of chemicals known to activate cannabinoid receptors in cells throughout the human body, including the skin. Phytocannabinoids are the cannabinoids derived from cannabis plants. They can be isolated from plants or produced synthetically. Endocannabinoids are endogenous cannabinoids produced naturally by cells in the human body. Canonical phytocannabinoids are ABC tricyclic terpenoid compounds bearing a benzopyran moiety.

[0113] Cannabinoids exert their effects by interacting with cannabinoid receptors present on the surface of the cells. To date, two types of cannabinoid receptor have been identified, the CB1 receptor and the CB2 receptor. These two receptors share about 48% amino acid sequence identity and are distributed in different tissues and have distinct cell signaling mechanisms. They also differ in their sensitivity to agonists and antagonists.

[0114] In some cases, a cannabinoid or precursor thereof, can be purified, derivatized (e.g., to form a prodrug or salt, or to form a target cannabinoid from the precursor), and / or formulated in a pharmaceutical composition.

[0115] Cannabinoids include but are not limited to phytocannabinoids. In some cases the cannabinoids include but are not limited to, cannabinol, cannabidiols, ^9-tetrahydrocannabinol ( ^9-THC), the non-natural cannabinoid HU-210 (6aR,10aR)-9-(hydroxymethyl)-6,6-dimethyl-3- (2-methyloctan-2-yl)-6H,6aH,7H,10H,10aH-benzo[c]isochromen-1-ol), HU-308 ([(1R,2R,5R)- 2-[2,6-dimethoxy-4-(2-methyloctan-2-yl)phenyl]-7,7-dimethyl-4-bicyclo[3.1. 1]hept-3- enyl]methanol), HU-433 an enantiomer of HU-308, cannabidivarin (CBDV), cannabichromene (CBC), cannabichromevarin (CBCV), cannabigerol (CBG), cannabigerovarin (CBGV), -27- 1101228610\3\AMERICASAttorney Docket No.120754.00100 cannabielsoin (CBE), cannabicyclol (CBL), cannabivarin (CBV), and cannabitriol (CBT). Still other cannabinoids include, including tetrahydrocannibivarin (THCV) and cannabigerol monomethyl ether (CBGM). Additional cannabinoids include cannabichromenic acid (CBCA), ^9-tetrahydrocannabinolic acid (THCA); and cannabidiolic acid (CBDA); these additional cannabinoids are characterized by the presence of a carboxylic acid group in their structure.

[0116] Still other cannabinoids include nabilone, rimonabant, JWH-018 (naphthalen-1-yl-(1- pentylindol-3-yl)methanone), JWH-073 naphthalen-1-yl-(1-butylindol-3-yl)methanone, CP-55940 (2-[(1R,2R,5R)-5-hydroxy-2-(3-hydroxypropyl) cyclohexyl]-5-(2-methyloctan-2- yl)phenol), dimethylheptylpyran, HU-331 (3-hydroxy-2-[(1R)-6-isopropenyl-3-methyl-cyclohex- 2-en-1-yl]-5-pentyl-1,4-benzoquinone), SR144528 (5-(4-chloro-3-methylphenyl)-1-[(4- methylphenyl)methyl]-N-[(1S,2S,4R)-1,3,3-trimethylbicyclo[2.2.1]heptan-2-yl]-1H-pyrazole-3- carboxamide), WIN 55,212-2 ((11R)-2-methyl-11-[(morpholin-4-yl)methyl]-3-(naphthalene-1- carbonyl)-9-oxa-1-azatricyclo[6.3.1.04,¹²]dodeca-2,4(12),5,7-tetraene), JWH-133 ((6aR,10aR)-3- (1,1-dimethylbutyl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6H-dibenzo[b,d]pyran), levonatradol, and AM-2201 (1-[(5-fluoropentyl)-1H-indol-3-yl]-(naphthalen-1-yl)methanone). Other cannabinoids include ^8-tetrahydrocannabinol ( ^8-THC), 11-hydroxy- ^9- tetrahydrocannabinol, ^11-tetrahydrocannabinol, and 11-hydroxy-tetracannabinol.

[0117] In another alternative, analogs or derivatives of these cannabinoids can be obtained by providing a precursor cannabinoid and further derivatization, e.g., by synthetic means. Non- natural cannabinoids include, but are not limited to, those described in United States Patent No. 9,394,267 to Attala et al.; United States Patent No.9,376,367 to Herkenroth et al.; United States Patent No.9,284,303 to Gijsen et al.; United States Patent No.9,173,867 to Travis; United States Patent No. 9,133,128 to Fulp et al.; United States Patent No. 8,778,950 to Jones et al.; United States Patent No. 7,700,634 to Adam-Worrall et al.; United States Patent No. 7,504,522 to Davidson et al.; United States Patent No. 7,294,645 to Barth et al.; United States Patent No. 7,109,216 to Kruse et al.; United States Patent No.6,825,209 to Thomas et al.; and United States Patent No.6,284,788 to Mittendorf et al.

[0118] In one or more embodiments, a compound of the present disclosure is a cannabinoid. Cannabinoids according to the present disclosure can be at least partially selective for binding to either the CB2 cannabinoid receptor or the CB1 cannabinoid receptor. In one or more embodiments, the cannabinoids bind both the CB1 and CB2 cannabinoid receptors. In some cases, cannabinoids according to the present disclosure are selective for the CB1 cannabinoid receptor and act as partial agonists. In some other cases, cannabinoids according to the present disclosure are selective for the CB2 cannabinoid receptor and act as partial agonists. In some cases, the -28- 1101228610\3\AMERICASAttorney Docket No.120754.00100 cannabinoids bind to both CB1 and CB2 receptors acting as partial agonists for both receptors but with higher affinity to CB2 receptor at a similar potency to that of ∆9-THC. In some cases, the cannabinoids, or one of the cannabinoids in a mixture of cannabinoids is an antagonist or inverse agonist of CB1 and / or CB2 receptor. As antagonists or inverse agonist, the cannabinoids can bind to the CB1 and / or CB2 receptor but can induce a pharmacological response opposite to that of agonist. In some cases, cannabinoids in the pharmaceutical compositions and methods according to the present disclosure are partially selective for the CB2 cannabinoid receptor. In some cases, the cannabinoid or mixture of cannabinoids may exhibit, e.g., at least, a 3-fold lower Kiagainst CB2 receptor as compared to CB1 receptor in an in vitro competition assay with an overall higher binding affinity to CB2. Pharmaceutical Compositions

[0119] The pharmaceutical compositions described herein are typically formulated for administration. Accordingly, described herein is a pharmaceutical composition comprising 4-chlorobutyl-CBN formulated for administration with one or more and a pharmaceutically acceptable carrier(s), diluent(s), or excipient(s).

[0120] The pharmaceutical compositions may be prepared by known procedures using well-known and readily available ingredients.

[0121] Pharmaceutical compositions comprising 4-chlorobutyl-CBN may be formulated for administration to a subject by one of a variety of standard routes, for example, ocularly, orally, topically, parenterally, by inhalation or spray, rectally or vaginally, in a dosage unit containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, excipients and / or vehicles.

[0122] The term parenteral as used herein includes in one or more embodiments subcutaneous injections, intradermal, intra-articular, intravenous, intramuscular, intravascular, intrasternal, intrathecal injection and infusion techniques. The pharmaceutical composition will typically be formulated in a format suitable for administration to the subject by the selected route, for example, as an eyedrop, an ocular ophthalmic depot, a syrup, elixir, tablet, troche, lozenge, hard or soft capsule, pill, suppository, oily or aqueous suspension, dispersible powder or granule, emulsion, injectable, or solution.

[0123] In one or more embodiments, the pharmaceutical composition is formulated for administration via a systemic route, for example, intravenously, intramuscularly, intradermally, intraperitoneally, subcutaneously, or orally.

[0124] Pharmaceutical compositions intended for oral use may be prepared in either solid or fluid unit dosage forms. Fluid unit dosage form can be prepared according to procedures known -29- 1101228610\3\AMERICASAttorney Docket No.120754.00100 in the art for the manufacture of pharmaceutical compositions and such pharmaceutical compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. An elixir can be prepared by using a hydroalcoholic (for example, ethanol) vehicle with suitable sweeteners such as sugar or saccharin, together with an aromatic flavoring agent, among other techniques for preparation. Suspensions can be prepared with an aqueous vehicle with the aid of a suspending agent such as acacia, tragacanth, methylcellulose and the like. In some embodiments, the pharmaceutical composition comprises a compound of Formula I or a derivative thereof; and / or a pharmaceutically acceptable salt thereof; and the pharmaceutically acceptable excipient comprises ethanol. In some embodiments, the pharmaceutical composition comprises a compound of Formula I or a derivative thereof; and / or a pharmaceutically acceptable salt thereof; and the pharmaceutically acceptable excipient does not comprise ethanol. In some embodiments, the pharmaceutically acceptable excipient consists solely of ethanol. In some embodiments, the pharmaceutically acceptable excipient does not consist solely of ethanol.

[0125] Solid pharmaceutical compositions including but not limited to tablets, gels, or chews contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of the same. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate: granulating and disintegrating agents for example, corn starch, or alginic acid: binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc and other conventional ingredients such as dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, methylcellulose, and functionally similar materials. The solid pharmaceutical compositions may be uncoated or they may be coated by known techniques. Among other reasons, a solid pharmaceutical composition that is coated such as a coated tablet can delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over an extended period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.

[0126] Pharmaceutical compositions for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, medium chain triglyceride oil (MCTs oil), medium chain fatty acids (MCFAs), liquid paraffin, coconut oil, palm kernel oil, olive oil, or in some cases peanut oil. Soft gelatin capsules are prepared by machine encapsulation of a slurry of -30- 1101228610\3\AMERICASAttorney Docket No.120754.00100 the compound with an acceptable inert oil, such as vegetable oil, light liquid petrolatum, MCTs oil, MCFAs, coconut oil, palm kernel oil, peanut oil, or other inert oil.

[0127] Aqueous suspensions contain the active ingredient in admixture with one or more excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, for example sodium carboxylmethylcellulose, methyl cellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; and dispersing or wetting agents such as naturally-occurring phosphatides (for example, lecithin), condensation products of an alkylene oxide with fatty acids (for example polyoxyethylene stearate), condensation products of ethylene oxide with long chain aliphatic alcohols (for example hepta-decaethyleneoxycetanol), condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol (for example, polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (for example polyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n-propyl-p-hydroxy benzoate, one or more coloring agents, one or more flavoring agents or one or more sweetening agents, such as sucrose or saccharin.

[0128] Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide palatable oral preparations. These pharmaceutical compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

[0129] Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.

[0130] Pharmaceutical compositions may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, for example olive oil or peanut oil, or a mineral oil, for example liquid paraffin, or mixtures of these. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of such partial esters with ethylene -31- 1101228610\3\AMERICASAttorney Docket No.120754.00100 oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also optionally contain sweetening and flavoring agents.

[0131] The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleaginous suspension. Such suspensions may be formulated as known in the art using suitable dispersing or wetting agents and suspending agents such as those mentioned above. The sterile injectable preparation may also be a sterile injectable solution or a suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Other acceptable vehicles and solvents that may be employed include, for example, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils may be employed as a solvent or suspending medium. Various bland fixed oils known to be suitable for this purpose may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Adjuvants such as local anesthetics, preservatives and buffering agents may also optionally be included in the injectable solution or suspension.

[0132] The pharmaceutical compositions may comprise a pharmaceutically acceptable excipient such as a buffer. In one or more embodiments, the buffer is a sterile buffer. In one or more embodiments, the buffer is a PBS buffer. In one or more embodiments, the buffer comprises a polysorbate-type nonionic surfactant, such as polysorbate 20 or polysorbate 80. In one or more embodiments, the polysorbate-type nonionic surfactant is present in an amount from 0.01 to 1% (w / v), such as 0.02 to 0.4% (w / v), or 0.03% (w / v) of the buffer. In one or more embodiments, the buffer comprises a sugar, such as sucrose. In one or more embodiments, the sugar is present in an amount from 1 to 10% (w / v), such as from 2 to 8% (w / v), 4 to 6% (w / v), or 5% (w / v) of the buffer. In one or more embodiments, the buffer comprises a solvent, such as dimethyl sulfoxide (DMSO). In one or more embodiments, the DMSO is present in an amount from 0.1 to 1% (w / v), such as from 0.1 to 0.5% (w / v) or 0.2 to 0.3% (w / v) of the buffer.

[0133] Other pharmaceutical compositions and methods of preparing pharmaceutical compositions are known in the art and are described, for example, in “Remington: The Science and Practice of Pharmacy” (formerly “Remington’s Pharmaceutical Sciences”); Gennaro, A., Lippincott, Williams & Wilkins, Philadelphia, Pa. (2000).

[0134] The concentration of the compound (e.g., 4-chlorobutyl-CBN) in the pharmaceutical composition will vary depending on the condition to be treated and / or the mode of administration. Methods

[0135] Described herein are methods of protecting a neuron from neurodegenerative stimuli. In general, the methods include contacting the neuron with an effective amount of 4-chlorobutyl- CBN or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising -32- 1101228610\3\AMERICASAttorney Docket No.120754.00100 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a 4-chlorobutyl-CBN derivative (e.g., a prodrug). The method can be an in vitro method. Alternatively, the method can be a method performed at least partially in vivo, such as by administering a compound or a pharmaceutical composition to a subject. The administering can be performed by systemic (e.g., intravenous, or subcutaneous), or localized injection. For example, localized injection can comprise intravitreal injection. The administering can be performed by a localized administration method that is non-invasive. For example, localized administration to retinal neurons, such as retinal ganglia can include administration of an eye drop pharmaceutical composition, such as a hydrogel (see, e.g., WO 2018 / 205022) or a microemulsion (see, e.g., U.S. Patent Nos.8,968,775 and 9,149,453).

[0136] As described in U.S. 8,968,775 and U.S. 9,149,453, an eye drop pharmaceutical composition may include: a pharmaceutical carrier suitable for topical administration to the eye, comprising: an oil-in-water microemulsion comprising (i) an oil selected from the group consisting of isopropyl myristate, isopropyl palmitate, and medium chain triglycerides; (ii) a pair of surfactants selected from the group consisting of two polysorbates, a polysorbate and propylene glycol, a polysorbate and glycerol, a polysorbate and 1,2,3-triacetoxypropane, polyethoxylated castor oil and 1,2,3-triacetoxypropane, and polyethoxylated castor oil and propylene glycol; and (iii) water, wherein: the water represents 50 to about 95 percent (w / w) of the pharmaceutical carrier, the oil and the pair of surfactants represent substantially all of the remainder of the pharmaceutical carrier; and the ratio of percent (w / w) the pair of surfactants to percent (w / w) the oil is at least about 10:1.

[0137] A pharmaceutical composition may comprise a lipophilic active pharmaceutical ingredient (API) and the above-described pharmaceutical carrier, wherein the pharmaceutical composition is formulated for topical administration to the eye.

[0138] In one or more embodiments, the eye drop pharmaceutical composition includes a compound of Formula I.

[0139] In one or more embodiments, the compound or pharmaceutical composition thereof is administered for a period of less than six weeks. In one or more embodiments, the compound or pharmaceutical composition thereof is administered for a period of about one to four weeks. In one or more embodiments, such as to treat a neurodegenerative disease such as age-related macular degeneration, the compound or pharmaceutical composition thereof will be administered for an extended period of time, such as for several years, or for the remaining life of the patient. The compound or pharmaceutical composition thereof may be administered bimonthly (every other month), monthly, weekly, every other day, daily, twice per day, or three times per day. -33- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0140] The compound or pharmaceutical composition thereof may be administered to treat an eye of a subject in need of treatment to protect retinal neurons (e.g., optical nerve fibers). For example, the subject may have age-related macular degeneration. In some or any embodiments, the subject may have glaucoma. In some or any embodiments, the subject does not have glaucoma. If the compound or pharmaceutical composition thereof is administered to protect neurons, such as retinal neurons, then the compound or pharmaceutical composition thereof can be administered at a dosage that provides a peak, median, or trough, preferably peak, neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells. In one or more embodiments, the target neuron is a retinal neuron. In one or more embodiments, the target neuron is a peripheral neuron. In one or more embodiments, the target neuron is a central neuron. In one or more embodiments, the compound or pharmaceutical composition thereof is administered to treat an eye of a subject in need of treatment to protect retinal tissue, such as the retina and / or retinal pigment epithelium (RPE). In one or more embodiments, the compound or pharmaceutical composition thereof is administered to treat an eye of a subject in need of treatment to protect photoreceptor cells (photoreceptors). In one or more embodiments, the compound or pharmaceutical composition thereof is administered to treat an eye of a subject in need of treatment to protect against accumulation of or reduction of autofluorescent extracellular debris. In one or more embodiments, the compound or pharmaceutical composition thereof is administered to treat an eye of a subject in need of treatment to protect against neuroinflammation (in some embodiments where the subject has AMD; in some embodiments where the subject has glaucoma; in some embodiments where the subject does not have glaucoma).

[0141] In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is less than about 25 µM, less than about 20 µM, less than about 15 µM, less than about 14 µM, less than about 13 µM, less than about 12 µM, or less than about 10 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is from greater than about 0.15 µM to less than about 25 µM, or from greater than 0.15 µM to less than 25 µM, or from at least about 0.15 µM to less than about 25 µM, or from at least 0.15 µM to less than 25 µM, or from greater than about 0.15 µM to less than about -34- 1101228610\3\AMERICASAttorney Docket No.120754.00100 20 µM, or from greater than 0.15 µM to less than 20 µM, or from at least about 0.15 µM to less than about 20 µM, or from at least 0.15 µM to less than 20 µM.

[0142] In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is from greater than about 0.15 µM to less than about 15 µM, or from greater than 0.15 µM to less than 15 µM, or from at least about 0.15 µM to less than about 15 µM, or from at least 0.15 µM to less than 15 µM, or from greater than about 0.15 µM to less than about 12 µM, or from greater than 0.15 µM to less than 12 µM, or from at least about 0.15 µM to less than about 12 µM, or from at least 0.15 µM to less than 12 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) is from at least about 0.5 µM to less than about 25 µM, or from at least 0.5 µM to less than 25 µM, or from at least about 0.5 µM to less than about 20 µM, or from at least 0.5 µM to less than 20 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) is from at least about 0.5 µM to less than about 15 µM, or from at least 0.5 µM to less than 15 µM, or from at least about 0.5 µM to less than about 12 µM, or from at least 0.5 µM to less than 12 µM.

[0143] In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is less than about 25 µM, less than about 20 µM, less than about 15 µM, less than about 14 µM, less than about 13 µM, less than about 12 µM, or less than about 10 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is from greater than about 0.15 µM to less than about 25 µM, or from greater than 0.15 µM to less than 25 µM, or from at least about 0.15 µM to less than about 25 µM, or from at least 0.15 µM to less than 25 µM, or from greater than about 0.15 µM to less than about 20 µM, or from greater than 0.15 µM to less than 20 µM, or from at least about 0.15 µM to less than about 20 µM, or from at least 0.15 µM to less than 20 µM.

[0144] In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative -35- 1101228610\3\AMERICASAttorney Docket No.120754.00100 (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is from greater than about 0.15 µM to less than about 15 µM, or from greater than 0.15 µM to less than 15 µM, or from at least about 0.15 µM to less than about 15 µM, or from at least 0.15 µM to less than 15 µM, or from greater than about 0.15 µM to less than about 12 µM, or from greater than 0.15 µM to less than 12 µM, or from at least about 0.15 µM to less than about 12 µM, or from at least 0.15 µM to less than 12 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) is from at least about 0.5 µM to less than about 25 µM, or from at least 0.5 µM to less than 25 µM, or from at least about 0.5 µM to less than about 20 µM, or from at least 0.5 µM to less than 20 µM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) is from at least about 0.5 µM to less than about 15 µM, or from at least 0.5 µM to less than 15 µM, or from at least about 0.5 µM to less than about 12 µM, or from at least 0.5 µM to less than 12 µM.

[0145] In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof or a derivative (e.g., a prodrug) thereof) in the vitreous humor, in the ocular tissues of the eye, and / or in contact with ocular neuronal cells is from greater than about 0.15 µM to less than about 10 µM, or from greater than 0.15 µM to less than 7.5 µM, or from at least about 0.15 µM to less than about 10 µM, or from at least 0.15 µM to less than 7.5 µM, or from greater than about 0.15 µM to about 5 µM, or from greater than 0.15 µM to 5 µM, or from at least about 0.15 µM to about 5 µM, or from at least 0.15 µM to 5 µM.

[0146] For example, the compound or pharmaceutical composition thereof can be administered orally, intrathecally, intravenously, topically, or injected, and / or administered directly to the site of the target neuron or target neuronal population. In one or more embodiments, the neuroprotective effective concentration is achieved by a systemic dosage of from about 1 mg / kg to about 100 mg / kg, from about 1 mg / kg to about 20 mg / kg, from about 1 mg / kg to about 15 mg / kg, from about 1 mg / kg to about 10 mg / kg, or from about 1 mg / kg to about 13 mg / kg. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day.

[0147] The compound or pharmaceutical composition thereof can be administered intrathecally, intravenously, or injected, or administered directly into the eye such as by topical eye instillation or intravitreal injection or pump. In one or more embodiments, for ocular administration in an ocular indication (e.g., to treat age-related macular degeneration), the -36- 1101228610\3\AMERICASAttorney Docket No.120754.00100 systemic dose can be from 1 mg / kg to 20 mg / kg. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day. In one or more embodiments, for systemic administration in an ocular indication (e.g., to treat age-related macular degeneration), the systemic dose can be from 1 mg / kg to 15 mg / kg, from 1 mg / kg to 13 mg / kg, or from 1 mg / kg to 10 mg / kg. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day. In one or more embodiments, for systemic administration in a peripheral indication (e.g., to treat peripheral neuropathy and / or peripheral nerve insult or injury), the dose can be from 1 mg / kg to 20 mg / kg, from 1 mg / kg to 15 mg / kg, from 1 mg / kg to 13 mg / kg, or from 1 mg / kg to 10 mg / kg. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day. In one or more embodiments, for systemic administration in a central indication (e.g., to treat a central nerve insult or injury), the dose can be from 1 mg / kg to 20 mg / kg, from 1 mg / kg to 15 mg / kg, from 1 mg / kg to 13 mg / kg, or from 1 mg / kg to 10 mg / kg. The dose can be repeated, e.g., weekly, daily, or twice a day.

[0148] In one or more embodiments, for ocular administration in an ocular indication (e.g., to treat age-related macular degeneration), the ocular dose can be from 0.5 mg to 20 mg, from 0.5 mg to 15 mg, from 0.5 mg to 10 mg, from 1 mg to 20 mg, from 1 mg to 15 mg, from 1 mg to 10 mg, from 0.5 mg to 5 mg, or from 1 mg to 5 mg applied to the eye, such as in the form of an eye drop or eye gel. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day. In one or more embodiments, for ocular administration in an ocular indication (e.g., to treat age-related macular degeneration), the ocular dose can be from 0.05 mg to 2 mg, from 0.05 mg to 1.5 mg, from 0.05 mg to 1 mg, from 0.1 mg to 2 mg, from 0.1 mg to 1.5 mg, from 0.1 mg to 1 mg, from 0.05 mg to 0.5 mg, or from 0.1 mg to 0.5 mg applied to the eye, such as in the form of an intravitreal injection or pump. The dose can be repeated, e.g., bimonthly, monthly, weekly, every other day, daily, or twice a day.

[0149] In one or more embodiments, the compound or pharmaceutical composition thereof is administered within about 0-48 hours of an insult affecting the retinal neurons, photoreceptors, retinal pigment epithelium, neuroinflammation, and / or accumulation of autofluorescent extracellular deposits. In one or more embodiments, the compound or pharmaceutical composition thereof is administered within about 2-24 hours of an insult affecting the retinal neurons, photoreceptors, and / or retinal pigment epithelium. In one or more embodiments, the compound or pharmaceutical composition thereof is administered within about 3-12 hours of an insult affecting the retinal neurons, photoreceptors, and / or retinal pigment epithelium. In one or more embodiments, the compound or pharmaceutical composition thereof is administered within about -37- 1101228610\3\AMERICASAttorney Docket No.120754.00100 3-5 hours of an insult affecting the retinal neurons, photoreceptors, and / or retinal pigment epithelium.

[0150] In one or more embodiments, the compound or pharmaceutical composition thereof is administered to a subject having AMD.

[0151] The method may include or further include administering an additional active agent (for example, a second drug) simultaneously or sequentially in combination with the compound or pharmaceutical composition thereof. In some cases, the additional active agent is a therapeutic for treatment of age-related macular degeneration. In one or more embodiments, an additional active agent is an anesthetic. In one or more embodiments, an additional active agent is a VEGF inhibitor. In one or more embodiments, an additional active agent is an anti-VEGF protein or compound. For example, the method may include anaesthetizing the subject prior to administering a compound or pharmaceutical composition of one or more embodiments. In another example, the subject is anaesthetized prior to the contacting (of the administering step) in the method of one or more embodiments. EXAMPLES Materials and Methods

[0152] Animals and experimental groups

[0153] Sprague Dawley (SD) albino rats were born and raised in dim cyclic light condition (12 hours light, 12 hours dark) with an ambient light level of approximately 5 lux. The animals had free access to food and water. The experiments were conducted on adult animals aged 2-4 months. The treatments were administered through intravitreal injection in both eyes the day before light damage and the possible retinal protection was assessed seven days after light damage through fERG recordings.

[0154] Preparation of the pharmaceutical composition and intravitreal injections

[0155] 4-chlorobutyl-CBN was prepared by a combination of biosynthesis and chemical synthesis. Briefly, 5-chlorovaleric acid was “fed” to a strain of Saccharomyces cerevisiae engineered to convert the acid analog to 4-chlorobutyl-olivetolic acid: . The 4-chlorobutyl-olivetolic acidthe culture medium by extraction with ethyl acetate. From 4-chlorobutyl-olivetolic acid, two different routes were used to generate -38- 1101228610\3\AMERICASAttorney Docket No.120754.00100 the 4-chlorobutyl-CBN final product. For route “A,” the 4-chlorobutyl-olivetolic acid was decarboxylated to 4-chlorobutyl-olivetol: and then combined with citral 4-chlorobutyl-CBC:which was oxidized to 4- , the 4-chlorobutylolivetolicacid was “fed” to a strain the geranyl-diphosphate precursor (GPP) and to express heterologous genes encoding a prenyltransferase and a synthase. The bioconversion strain converted the 4-chlorobutyl-olivetolic acid analog to 4-chlorobutyl- THCA: which was recovered from theThe 4-chlorobutyl-THCA was then decarboxylated and oxidized to generate 4-chlorobutyl-CBN. 4-chlorobutyl-CBN can also be prepared by chemical synthesis. In addition, methods similar to those described in US- 2023-0063396-A1 are or can be used in the preparation of 4-chlorobutyl-CBN.

[0156] A 15 mM or 30 mM stock solution of 4-Cl-butyl CBN in 100% DMSO was diluted in sterile dilution buffer (0.03% polysorbate (Tween) 20, 5% sucrose in PBS) to attain a final formulation concentration of ~100 µM, 30 µM, and 10 µM at 0.2-0.4% final DMSO concentration for intravitreal injection administration. Formulations were prepared by serial dilution, and it was ensured that similar levels of DMSO were included at the different dose levels. The buffer solution was prepared by dissolving 0.03% polysorbate (tween) 20, 5% sucrose in PBS buffer. For buffer solutions used in vehicle(s) and control(s) (or injection formulations), the buffer solution was diluted with DMSO at the appropriate level (to match what was used in the stock solution(s) of the corresponding concentration / dilution, except without the cannabinoid / API). Regarding DMSO in buffer solution, the injection formulation(s) included DMSO from the 4-chlorobutyl- CBN stock solution, e.g., where the stock was diluted to 100 µM when injected in the eye, the 4- chlorobutyl-CBN concentration was 10 µM. Accordingly, DMSO was added in the buffer solution -39- 1101228610\3\AMERICASAttorney Docket No.120754.00100 for vehicle and control experiments to match the DMSO concentration of the corresponding injection formulation(s). The buffer solution was used in the vehicle(s) and control(s), to be described. After dissolving the excipients, the buffer was filtered through a 0.2 µM filter to prepare sterile buffer for formulation preparation. DMSO stock solution was diluted in the buffer solution to achieve ~100 µM, 30 µM, and 10 µM concentrations of final formulation. This was followed by vortex mixing for 30 seconds or more on the high setting. The final concentration in the vitreous humor was 1 / 10 of the injected concentration. For example, an initial concentration of 100 µM at 1 / 10 was a final concentration of 10 µM (see figures); likewise, an initial concentration of 30 µM at 1 / 10 was a final concentration of 3 µM (see figures); further, an initial concentration of 3 µM at 1 / 10 was a final concentration of 0.3 µM (see figures); and so on. The final projected concentration achieved inside the eye for the buffer components was 0.003% polysorbate (tween) 20, 0.5% sucrose, and ~0.02% to 0.04% DMSO.

[0157] For the surgical procedure, rats were anaesthetized with an intraperitoneal injection of Ketamine / Xylazine (10 mg / 100 g–1.2 mg / 100 g).2.0 μL of the compounds were injected in both eyes using a Hamilton syringe under total sterile conditions (2). Control: Healthy animals with no light damage (LD) and no treatment. Vehicle: light damage and treatment with dilution buffer. The vehicle did not induce any change of the fERG responses compared to light damage un- injected eyes. Therefore, the light damage group was not included in the analysis.

[0158] Light damage

[0159] Animals were placed in individual Plexiglas cages with food placed on the floor and water in plastic bottles. They were dark adapted overnight and exposed to 1000 lux at 9 a.m. for 24 h (LD24h). Thereafter, they were returned to dim cyclic light conditions for 7 days to recover from the acute stress due to light exposure.

[0160] Electroretinogram recordings

[0161] The fERG was recorded in a dark-adapted condition in response to a single white light flash of increasing intensity (scotopic), delivered by a standard Ganzfeld Stimulator (Biomedica Mangoni, Pisa, Italy). Before performing fERG, the rats were anaesthetized with an intraperitoneal injection of Ketamine / Xylazine (10 mg / 100 g–1.2 mg / 100 g), mounted on a stereotaxic apparatus and the body temperature was maintained at 37.5°C. Corneas were anaesthetized with a drop of procaine (NOVOCAINE) and pupils were dilated with visumidriatic 1.0% tropicamide. The recordings were carried out for both eyes simultaneously with a gold electrode loop (2.0 mm in diameter) placed on the corneas. The reference electrodes were inserted subcutaneously in the proximity of the eyes and the ground electrode was inserted in the anterior scalp, between the eyes. The responses were recorded at increasing light intensities (0.001 cd*s / m2 to 100 cd*s / m2 -40- 1101228610\3\AMERICASAttorney Docket No.120754.00100 range). At the end of the recording session, traces were bandpass filtered between 0.3 and 500 Hz. The a-wave amplitude (μV) and b-wave amplitude (μV) for the respective luminance was measured. Oscillatory potentials (OPs) analysis was also performed by calculating the sum of the amplitude of OP1, OP2, OP3 and OP4.

[0162] Statistical analysis

[0163] For statistical analysis the one-way ANOVA test followed by Dunnett’s post hoc comparison was performed between the CBN, 4-chlorobutyl-CBN, and Vehicle groups; the Student’s t-test was performed between 4-chlorobutyl-CBN and Vehicle groups for the analysis where the CBN group was not included. The statistical analysis was conducted by using the SigmaPlot 12.0 software (Systat Software, San Jose, CA, USA). Data are shown as mean ± standard error of mean. (4-Cl-CBN: n=14; CBN 30: n=17; CBN 100: n=13; vehicle: n=17) (*p<0.05 versus vehicle).

[0164] Example 1: in vivo intravitreal injections, 4-chlorobutyl-CBN at 0.3 µM

[0165] An in vivo study was performed in an AMD rat model via intravitreal injection of 3 µM (0.3 µM final concentration inside the eye) of 4-chlorobutyl-CBN in vehicle and 3 µM (0.3 µM final concentration inside the eye) of CBN in vehicle, in separate experiments. The vehicle alone and control were tested and compared separately. Intravitreal injection was performed at the back of the eye, as shown in FIG.1 (arrow). The results of this study are shown in FIGS.2A and 2B.

[0166] As can be seen in FIGS. 2A and 2B, CBN at 0.3 µM final concentration appeared to outperform 4-chlorobutyl-CBN at 0.3 µM final concentration. However, the result of the Student’s t-test between CBN and 4-Cl-CBN groups is p=0.46; hence, there is no statistically significant difference between the two groups. In this study, the 4-chlorobutyl-CBN at 0.3 µM final concentration was tested in a smaller number of animals. Thus, these results may not be statistically significant.

[0167] Example 2: in vivo intravitreal injections, 4-chlorobutyl-CBN at 3 µM

[0168] An in vivo study was performed in an AMD rat model via intravitreal injection of 30 µM (3 µM final) of 4-chlorobutyl-CBN, 30 µM (3 µM final) of CBN, and 100 µM (10 µM final) of CBN, in separate experiments. The vehicle alone and control were tested and compared separately. Intravitreal injection was performed at the back of the eye, as shown in FIG.1 (arrow). The results of this study are shown in FIGS. 3A to 3F (a-wave amplitude), 4A to 4E (b-wave amplitude), and 5A (OPs amplitude).

[0169] The a-wave amplitude indicates the response of photoreceptors to light stimuli. As shown in FIGS.3A to 3F, 3 µM (final concentration) of 4-chlorobutyl-CBN outperformed 3 µM (final concentration) of CBN and 10 µM (final concentration) of CBN at luminance (light -41- 1101228610\3\AMERICASAttorney Docket No.120754.00100 intensities) of 0.01, 0.1, 1, 10, and 100 cd*s / m^2, for a-wave amplitude (µV) increase. From these results, statistically significant a-wave amplitude increases were seen at intensities of 0.01, 0.1, and 100 cd*s / m^2.

[0170] The b-wave amplitude indicated the overall response of the inner retina at increasing light stimuli. As shown in FIGS. 4A to 4E, 3 µM (final concentration) of 4-chlorobutyl-CBN outperformed 3 µM (final concentration) of CBN and 10 µM (final concentration) of CBN at luminance (light intensities) of 0.001, 0.01, 0.1, 1, 10, and 100 cd*s / m^2, for b-wave amplitude (µV) increases. From these results, statistically significant b-wave amplitude increases were seen at intensities of 0.1, 1, 10, and 100 cd*s / m^2.

[0171] The oscillatory potentials (OPs) amplitude indicated the response of the inner retinal circuitry to light stimuli (mainly due to rod bipolar and A11 / A17 amacrine cells). As shown in FIG.5, 3 µM (final concentration) of 4-chlorobutyl-CBN outperformed 3 µM (final concentration) of CBN and 10 µM (final concentration) of CBN at luminance (light intensities) of 0.001, 0.01, 0.1, 1, and 10 cd*s / m^2, for OPs amplitude (µV).

[0172] Example 3: Imaging

[0173] At the end of the electroretinogram recordings, the eyes were enucleated for morphological analyses, fixed in 4% paraformaldehyde for 6 h, and washed in 0.1 M phosphate buffered saline (PBS, pH 7.4). The cornea and the lens were removed, and the remaining eye cups were immersed in 30% sucrose for cryoprotection. They were then embedded in the Tissue Tek OCT (optimum cutting temperature, Qiagen, Valencia, CA, USA) compound, frozen and stored at -20°C. Cryosections of 20 μm thickness were then obtained through a Leica CM1850 cryostat (GmbH, Nussloch, Germany) and collected on gelatin and poly-l-lysine-coated slides. The sections crossing the optic nerve were selected for subsequent immunofluorescence analysis.

[0174] Immunofluorescence was used to identify microglia, astrocytes, and Müller cells. Specifically, non-specific binding sites were blocked using 5% bovine serum albumin (BSA) for 1 h at room temperature (R.T.). Sections were then incubated overnight at 4°C with primary antibodies (for detection of microglia (anti-IBA-1) (Wako Chemicals, #019-19741), astrocytes and Müller cells (anti-GFAP) (Cell Signaling, #3670). Sections were then incubated with secondary antibodies (anti-mouse or anti-rabbit IgG conjugated to red or green, fluorescent dies) (Alexa Fluor 594 or 488; Molecular Probes, Invitrogen, Carlsbad, CA, USA), diluted 1:300, and incubated at 37°C for 2 h. Bisbenzimide nuclear dye (Hoechst) was used to label the nuclei and identify the retinal layers. The images were then acquired by a fluorescence microscopy. Results are shown in FIG.6A. A negative image of the same is provided in FIG.6B to show contrast. -42- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0175] The light damage model of AMD was characterized by the thinning of the outer nuclear layer, where photoreceptors are located. In general, this is a characteristic of photoreceptors degeneration related to AMD. Alongside photoreceptor death, gliotic events develop in the retina, leading to (i) the invasion of the outer nuclear layer by microglia cells (that are present in the inner retina under physiological conditions), and (ii) reactivity of astrocytes and Müller cells characterized by the up regulation of the GFAP marker.

[0176] On this basis, the analysis on retinal cryosections investigated whether the abovementioned detrimental events were prevented by the administration of 4-Cl-Butyl CBN at 3 µM. Overall, the morphological analysis indicated that 4-Cl-Butyl CBN was able to prevent photoreceptors death and gliosis in the light damage retinas.

[0177] Specifically, the treatment provided the following protective outcomes: (i) an increased outer nuclear layer thickness compared to the vehicle; (ii) the inhibition of IBA-1 (+) cells invasion in the outer nuclear layer; and (iii) the dampening of GFAP expression throughout the retinal layers compared to the vehicle.

[0178] Example 4. Comparison between 4-chlorobutyl-CBN and control animals

[0179] To better evaluate the degree of functional protection carried out by 4-chlorobutyl-CBN in the light damage model, a Student’s t-test analysis was performed between the 4-chlorobutyl-CBN treated eyes and control (eyes of healthy animals). As shown in Table 12 of 18 of the fERG conditions investigated were not significantly different between control and treated eyes. Table 1: t-test comparison between 4-chlorobutyl-CBN and control animals t-test: 4-chlorobutyl-CBN versus control= Not statistically significant. The results indicate a trend that 4-chlorobutyl-CBN (such as at 3 µM final concentration as described above) significantly prevented retinal functional impairment due to light damage, since a-wave, b-wave, and oscillatory potentials (OPs) amplitudes compared to the vehicle. In general, 4-chlorobutyl-CBN preserved the fERG with values similar to the healthy undamaged animals. The functional protection by 4-chlorobutyl-CBN was evident for the b-wave -43- 1101228610\3\AMERICASAttorney Docket No.120754.00100 and OPs, suggesting greater protection of the inner retina compared to the outer retina. Additionally, a slight improvement of retinal function occurred also in response to CBN (when used at 3 µM) at 100 cd*s / m^2. In other words, in these tests there was little to no difference between the control (healthy eyes, not exposed to light) and the eyes treated with 4-chlorobutyl- CBN when exposed to light.

[0181] Example 5: in vivo concentration

[0182] 4-Cl-Butyl-CBN was tested at concentrations of 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). The a-wave amplitude, which indicated the response of the photoreceptors, was improved by the 4-Cl-Butyl-CBN at the concentrations of 3 µM and 10 µM compared to the vehicle, indicating that the treatment exerted neuroprotection of photoreceptors against light damage. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer. The results of these tests are shown in FIG.7A.

[0183] 4-Cl-Butyl-CBN was tested of 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). The b-wave amplitude, which indicated the overall response of the inner retina following light stimulation, was improved by the 4-Cl-Butyl-CBN at the concentrations of 3 µM and 10 µM compared to the vehicle, indicating that the treatment prevented retinal functional impairment. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer. The results of these tests are shown in FIG.7B.

[0184] 4-Cl-Butyl-CBN was tested at concentrations of 0.3 µM, 1 µM, 3 µM, and 10 µM (inside the eye). The oscillatory potentials (OPs) amplitude, which indicated the function of the inner retinal circuitry, was improved by the 4-Cl-Butyl-CBN at the concentrations of 3 µM and 10 µM compared to the vehicle, indicating that the treatment exerted retinal neuroprotection and prevented the inner retinal function. Control: Healthy animals with no light damage and no treatment. Vehicle: light damage and treatment with dilution buffer. The results of these tests are shown in FIG.7C.

[0185] The electrophysiological results obtained with light damage animals treated with 4-Cl Butyl CBN and CBN via intravitreal injection demonstrated that 4-Cl Butyl CBN at the concentration of 3 µM outperformed functional protection compared to vehicle-injected eyes, which is greater than that of CBN at the same concentration, as seen at least in FIGS.3A and 4A.

[0186] Example 6: Histology and molecular analysis

[0187] Representative retinal sections from all experimental group were stained with nuclear dye Hoechst (blue) as seen in FIG. 8A. The Control group refers to healthy rats without light -44- 1101228610\3\AMERICASAttorney Docket No.120754.00100 damage. The Vehicle group refers to light damage (diseased) animal and treated with injection buffer.

[0188] The red lines highlight the outer nuclear layer thickness (ONL). The ratio of ONL to retina thickness was determined as seen in FIG. 8B where FIG. 8C shows the specific locations on the superior retina and inferior retina where the thickness ratios were determined for FIG.8B. These data show that 4-Cl-Butyl-CBN improved outer nuclear layer thickness in an AMD model.

[0189] Autofluorescent (AF) extracellular deposits were assessed for retinal cryosections by using 594 nm excitation wavelength as seen in FIG.9A (AF deposits shown in red). The number of AF deposits was counted throughout the retinal sections as seen in FIG. 9B. These data show that 4-Cl-Butyl-CBN reduced extracellular deposits in an AMD model.

[0190] Retinal pigment epithelium (RPE) integrity was assessed in retinal cryosections immunostained with anti-RPE65 as seen in FIG. 10A (immunostaining shown in red). The cryosections crossing the optic nerve were selected for the analysis and the integrity of the RPE was measured at the dorsal retina. The white segmented lines delineate the intact RPE. RPE65 (+) length was measured as seen in FIG.10B. These data show that 4-Cl-Butyl-CBN improved RPE length in an AMD model.

[0191] Light damage models indicate loss of RPE integrity, including loss of RPE cell-cell connection integrity (Miralles de Imperial-Ollero, et al.2021, Cachafeiro, et al.2013 and Geiger, et al.2015).

[0192] Microglia were labeled with anti-IBA-1 (green) in retinal cryosections from all experimental groups as seen in FIG. 11A. IBA-1 (+) cells were counted at the outer retina (subretina, outer nuclear layer, outer plexiform layer) from the superior to the inferior edge of the retina as seen in FIG. 11B. Locations on the superior and inferior retina where the data were collected are the same as shown in FIG.8C. The numbers of IBA-1 (+) cells in the superior retina, inferior retina, and whole retina (superior and inferior) are shown in FIGS.11C-11E, respectively. These data show that 4-Cl-Butyl-CBN reduced inflammation in the retina in an AMD model.

[0193] Example 7: Receptor interaction studies

[0194] The interaction of 4-Cl-Butyl-CBN with different receptors was assessed by various assay methods. Assays were performed by Eurofins using their standard bioassay protocols.

[0195] The interaction of 4-Cl-Butyl-CBN with the CB1 and CB2 receptors using cAMP as an assay marker was assessed using the HitHunter(R) cAMP assay. Briefly, cAMP Hunter cell lines expressing either the CB1 or the CB2 receptor were assessed via the DiscoverX HitHunter cAMP XS+ assay. To assess agonist behavior (Gs agonist format), cells were incubated with sample to induce a response, media was aspirated and replaced with HBSS / Hepes buffer containing cAMP -45- 1101228610\3\AMERICASAttorney Docket No.120754.00100 XS+ Ab reagent, then 5 µL of 4X sample was added to the cells and incubated at 37oC or room temperature for 30 or 60 minutes with a vehicle concentration of 1%. To assess agonist behavior (Gi agonist format), cells were incubated in the presence of EC80 forskolin to induce a response, media was aspirated and replaced with HBSS / Hepes buffer containing cAMP XS+ Ab reagent, then 5 µL of 4X sample with 4X EC80 forskolin was added to the cells and incubated at 37oC or room temperature for 30 or 60 minutes with a vehicle concentration of 1%. To assess antagonist behavior, cells were pre-incubated with sample followed by an agonist challenge at the EC80 concentration, media was aspirated and replaced with HBSS / Hepes buffer containing cAMP CS+ Ab reagent, then 5 µL of 4X of the sample compound was added to the cells and incubated at 37oC or room temperature for 30 minutes, followed by addition of 5 µL of EC80 agonist and incubation at 37oC or room temperature for 30 or 60 minutes with a vehicle concentration of 1%. For GI coupled GPCRs, EC80 forskolin was included in the antagonist assay. For signal detection for either assay, 20 µL of cAMP XS+ ED / CL lysis cocktail was added and incubated for one hour followed by addition of 20 µL cAMP CS+ EA reagent and incubation at room temperature for 3 hours. Microplates were read for chemiluminescent detection.

[0196] The interaction of 4-Cl-Butyl-CBN with the CB1 and CB2 receptors using Arrestin as an assay marker was assessed using the PathHunter(R) β-Arrestin assay from Eurofins DiscoverX Corp.

[0197] The interaction of 4-Cl-Butyl-CBN with PPARγ, PPARα and PPARδ was assessed using the PathHunter(R) Nuclear Hormone Receptor Protein Interaction (Pro) assay. Briefly, for agonist determination, PathHunter NHR cell lines were incubated with sample, then 5 µL of 5X sample was added to the cells and incubated at 37oC or room temperature for 3-16 hours with a final vehicle assay concentration of 1%. For antagonist determination, cells were pre-incubated with antagonist followed by agonist challenge at the EC80 concentration, then 5 µL of 5X sample was added to the cells and incubated at 37oC or room temperature for 60 minutes with a final vehicle assay concentration of 1%, followed by addition of 5 µL of EC80 agonist in assay buffer and incubation at 37oC or room temperature for 3-16 hours. Assay signal was generated by addition of 12.5 or 15 µL of PathHunter detection reagent cocktail and incubation for one hour at room temperature followed by microplate reading for chemiluminescent detection.

[0198] Similar analyses were performed for the GPR55 receptor β-arrestin pathway.

[0199] Data on receptor interactions are provided in Table 2 below: Table 2: 4-Cl-Butyl-CBN Receptor Interaction Data Receptor Assay Type Agonist EC50 Antagonist IC501101228610\3\AMERICASAttorney Docket No.120754.00100 CB1 cAMP 2.65 none CB2 cAMP 6.37 none [002nist and antagonist and GPR55 agonist. These data further show that 4-Cl-Butyl-CBN is an agonist for the cAMP pathway and an antagonist for the β-arrestin pathway.

[0201] Example 8: 4-Cl-Butyl-CBN levels in retina following topical and IVT deliveries

[0202] Topical eye drop formulations of 4-Cl-Butyl-CBN were administered topically to rabbits in two formulations and compared to IVT levels in rats as seen in FIG.13.

[0203] IVT delivery in rats was performed in the disease model as has been previously described in Example 1. 24 hours following IVT injection, animals were euthanized, and retina / RPE / choroid tissue was collected from each animal. The compound concentration was measured using qualified LC-MS / MS bioanalytical methods. The microemulsion formulation for the topical delivery in rabbits was based on previously described methods in U.S. Patent Nos. 8,968,775 and 9,149,453. In the topical delivery study performed n rabbits, the animals received two 40 µL topical doses (1% formulation) twice daily in both eyes for 8 days. On day 8, animals were euthanized, and retina / RPE / choroid tissue was collected from each animal. The compound concentration was measured using qualified LC-MS / MS bioanalytical methods.

[0204] Example 9: Topical delivery of 4-Cl-Butyl-CBN to retina in Sprague Dawley rat model

[0205] Topical administration of the compound is performed on Sprague Dawley albino rats via instillation of eye drops (10 µl) twice a day (~ 12 hours away) for seven days before light damage and will continue an additional 7 days after the damage. At the end point, retinal function is assessed through fERG recordings and RPE / retinal protection is investigated through histological evaluation of: RPE integrity, accumulation of autofluorescent deposits and retinal thickness.

[0206] Example 10: Comparative analysis of in vivo treatment of AMD with 4-Cl Butyl CBN versus CBN -47- 1101228610\3\AMERICASAttorney Docket No.120754.00100

[0207] Ocular tissues are taken from either the 4-Cl Butyl CBN or the CBN-treated rats for further morphological and molecular analysis on AMD pathology, described as follows.

[0208] 1. Retinal pigment epithelium (RPE) protection

[0209] Retinal pigment epithelium cell death is a component of AMD pathogenesis (Kim et al., 2021) and it is reproduced in the light damage model as well (Jaadane et al., 2017; Tisi et al., 2020a). The efficacy of the 4-Cl Butyl CBN or the CBN compounds to prevent retinal pigment epithelium degeneration is assessed on retinal cryosections immunolabeled with a selective RPE cell marker (RPE65), which allows evaluation of the maintenance of retinal pigment epithelium layer integrity by the treatment.

[0210] 2. Photoreceptors (PRs) neuroprotection

[0211] Photoreceptors cell death is another event in AMD (Mitchell et al., 2018) and is induced by light damage in albino rats as well. Photoreceptors neuroprotection is investigated by quantifying the outer nuclear layer thickness and by measuring the length of the degenerated area on retinal cryosections stained with bisbenzimide nuclear dye (Tisi et al., 2019). A greater outer nuclear layer thickness and a reduced length of the degenerated area is indicative of photoreceptors protection.

[0212] 3. Accumulation of extracellular deposits

[0213] Another AMD event is the accumulation of extracellular material (i.e., drusen and lipofuscin) that is visible through ocular fundus imaging (Ach et al., 2014; Davis et al., 2005). Also, in the light damage model, autofluorescent (AF) extracellular debris accumulates alongside retinal pigment epithelium and photoreceptors degeneration (Tisi et al., 2020b). Thus, autofluorescent extracellular debris is quantified on retinal cryosections through fluorescence microscopy, and a reduction of autofluorescent extracellular debris deposits in treated eyes is indicative of retinal protection.

[0214] 4. Neuroinflammation

[0215] Neuroinflammatory events are drivers of AMD. Retinal neuroinflammation relates to microglia and macroglia (astrocytes and Müller cells) reactivity (Gupta et al., 2003; Dhodapkar et al., 2022). Glial markers (IBA-1 for microglia, and GFAP for astrocytes and Müller cells) are used to investigate the neuroinflammatory events in terms of IBA-1 (+) cells and GFAP expression throughout the retinal layers. The reduction of the number of microglia cells and of the expression of GFAP in the light damage retina are indicative of reduced neuroinflammation.

[0216] The foregoing data further demonstrate neuroprotection of photoreceptors and improved photoreceptor function, reduced exocellular AF deposits and preserved RPE integrity in a light damaged in vivo rat model. In addition, 4-Cl-Butyl-CBN represents a compound with -48- 1101228610\3\AMERICASAttorney Docket No.120754.00100 systemic across the blood-brain-barrier and can be delivered via topical eye drop formulation or IVT formulation.

[0217] References Ramin, et al. “Age-Related Macular Degeneration: A Scientometric Analysis,” Med Hypothesis Discov. Innov. Ophthalmol.2015 Summer; 4(2): 39-49. Apte, “Age-Related Macular Degeneration,” N. Eng. J. Med.2021 Aug.5; 385(6): 539-547. Ach T, Huisingh C, McGwin G Jr, Messinger JD, Zhang T, Bentley MJ, Gutierrez DB, Ablonczy Z, Smith RT, Sloan KR, Curcio CA. Quantitative autofluorescence and cell density maps of the human retinal pigment epithelium. Invest Ophthalmol Vis Sci.2014 Jul 17;55(8):4832-41. doi: 10.1167 / iovs.14-14802. PMID: 25034602; PMCID: PMC4123894. Davis MD, Gangnon RE, Lee LY, Hubbard LD, Klein BE, Klein R, Ferris FL, Bressler SB, Milton RC; Age-Related Eye Disease Study Group. The Age-Related Eye Disease Study severity scale for age-related macular degeneration: AREDS Report No. 17. Arch Ophthalmol. 2005 Nov;123(11):1484-98. doi: 10.1001 / archopht.123.11.1484. Erratum in: Arch Ophthalmol. 2006 Feb;124(2):289-90. PMID: 16286610; PMCID: PMC1472813. Dhodapkar RM, Martell D, Hafler BP. Glial-mediated neuroinflammatory mechanisms in age- related macular degeneration. Semin Immunopathol. 2022 Sep;44(5):673-683. doi: 10.1007 / s00281-022-00939-3. Epub 2022 May 5. PMID: 35513496. Gupta N, Brown KE, Milam AH. Activated microglia in human retinitis pigmentosa, late-onset retinal degeneration, and age-related macular degeneration. Exp Eye Res. 2003 Apr;76(4):463-71. doi: 10.1016 / s0014-4835(02)00332-9. PMID: 12634111. Jaadane I, Villalpando Rodriguez GE, Boulenguez P, Chahory S, Carré S, Savoldelli M, Jonet L, Behar-Cohen F, Martinsons C, Torriglia A. Effects of white light-emitting diode (LED) exposure on retinal pigment epithelium in vivo. J Cell Mol Med.2017 Dec;21(12):3453-3466. doi: 10.1111 / jcmm.13255. Epub 2017 Jun 29. PMID: 28661040; PMCID: PMC5706508. Kim J, Lee YJ, Won JY. Molecular Mechanisms of Retinal Pigment Epithelium Dysfunction in Age-Related Macular Degeneration. Int J Mol Sci. 2021 Nov 14;22(22):12298. doi: 10.3390 / ijms222212298. PMID: 34830181; PMCID: PMC8624542. Mitchell P, Liew G, Gopinath B, Wong TY. Age-related macular degeneration. Lancet.2018 Sep 29;392(10153):1147-1159. doi: 10.1016 / S0140-6736(18)31550-2. PMID: 30303083. Ricci F, Bandello F, Navarra P, Staurenghi G, Stumpp M, Zarbin M. Neovascular Age-Related Macular Degeneration: Therapeutic Management and New-Upcoming Approaches. Int J Mol Sci. 2020 Nov 3;21(21):8242. doi: 10.3390 / ijms21218242. PMID: 33153227; PMCID: PMC7662479. -49- 1101228610\3\AMERICASAttorney Docket No.120754.00100 Tisi A, Flati V, Delle Monache S, Lozzi L, Passacantando M, Maccarone R. Nanoceria Particles Are an Eligible Candidate to Prevent Age-Related Macular Degeneration by Inhibiting Retinal Pigment Epithelium Cell Death and Autophagy Alterations. Cells.2020a Jul 4;9(7):1617. doi: 10.3390 / cells9071617. PMID: 32635502; PMCID: PMC7408137. Tisi A, Parete G, Flati V, Maccarone R. Up-regulation of pro-angiogenic pathways and induction of neovascularization by an acute retinal light damage. Sci Rep. 2020c Apr 14;10(1):6376. doi: 10.1038 / s41598-020-63449-y. PMID: 32286488; PMCID: PMC7156521. Tisi A, Passacantando M, Lozzi L, Maccarone R. Cerium oxide nanoparticles reduce the accumulation of autofluorescent deposits in light-induced retinal degeneration: Insights for age-related macular degeneration. Exp Eye Res. 2020b Oct;199:108169. doi: 10.1016 / j.exer.2020.108169. Epub 2020 Aug 3. PMID: 32758489. Tisi A, Passacantando M, Lozzi L, Riccitelli S, Bisti S, Maccarone R. Retinal long-term neuroprotection by Cerium Oxide nanoparticles after an acute damage induced by high intensity light exposure. Exp Eye Res.2019 May;182:30-38. doi: 10.1016 / j.exer.2019.03.003. Epub 2019 Mar 10. PMID: 30867118. Miralles de Imperial-Ollero, J., Gallego-Ortega, A., Norte-Munoz, M., Di Pierdomenico, J., Valiente-Soriano, F., Vidal-Sanz, M. An in vivo model of focal light emitting diode-induced cone photoreceptor phototoxicity in adult pigmented mice: Protection with bFGF. Exp. Eye Res.2021 Aug.;211:108746. Cachafeiro, M., Bemelmans, A-P., Smardzija, M., Afansieva, T., Pournaras, J-A., Grimm, C., Kostic, C., Phillippe, S., Wenzel, A., Arsenijevic, Y. Hyperactivation of retina by light in mice leads to photoreceptor cell death mediated by VEGF and retinal pigment epithelium permeability. Cell Death and Disease 2013;4:e781. Geiger, P., Barben, M., Grimm, C., Samardzija, M. Blue light-induced retinal lesions, intraretinal vascular leakage and edema formation in the all-cone mouse retina. Cell Death and Disease 2015;6:e1985. * * *

[0218] The inventions illustratively described herein can suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the future shown and described or any portion thereof, -50- 1101228610\3\AMERICASAttorney Docket No.120754.00100 and it is recognized that various modifications are possible within the scope of the invention claimed.

[0219] Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the inventions herein disclosed can be resorted by those skilled in the art, and that such modifications and variations are considered to be within the scope of the inventions disclosed herein. The inventions have been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the scope of the generic disclosure also form part of these inventions. This includes the generic description of each invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised materials specifically resided therein.

[0220] In addition, where features or aspects of an invention are described in terms of the Markush group, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. It is also to be understood that the above description is intended to be illustrative and not restrictive. Many embodiments will be apparent to those of in the art upon reviewing the above description. The scope of the invention should therefore, be determined not with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. The disclosures of all articles and references, including patent publications, are incorporated herein by reference. -51- 1101228610\3\AMERICAS

Claims

Attorney Docket No.120754.00100 CLAIMS What is claimed is:

1. A compound of Formula I: or a derivative thereof and / or asalt thereof.

2. The compound of claim 1, according to: .

3. A pharmaceuticalof claim 1 or 2, and a pharmaceutically acceptable excipient.

4. The pharmaceutical composition according to claim 3, in an amount (or concentration) sufficient to inhibit neurodegeneration in a subject, where the amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect retinal neurons, protect retinal tissue, protect photoreceptor cells, protect against accumulation of or reduce autofluorescent extracellular debris and protect against neuroinflammation.

5. The pharmaceutical composition according to claim 3 or 4, wherein the compound of Formula I is present in an amount from about 0.3 µM to less than or equal to about 20 µM; in some embodiments, from about 2 µM to less than or equal to about 12 µM; in some embodiments, from about 3 µM to less than or equal to about 10 µM; in some embodiments, from about 3 µM to less than or equal to about 15 µM; or in some embodiments, from about 10 µM to less than or equal to about 15 µM.

6. The pharmaceutical composition according to any one of claims 3 to 5, wherein the pharmaceutically acceptable excipient comprises a sterile buffer.

7. The pharmaceutical composition according to claim 6, wherein the sterile buffer is a PBS buffer or saline. -52- 1101228610\3\AMERICASAttorney Docket No.120754.00100 8. The pharmaceutical composition according to claim 6 or 7, wherein the sterile buffer comprises a polysorbate-type nonionic surfactant and sucrose.

9. The pharmaceutical composition according to claim 8, wherein the polysorbate-type nonionic surfactant is polysorbate 20.

10. The pharmaceutical composition according to any one of claims 3 to 9, comprising: a delivery carrier comprising a cellulosic polymer and an anionic polysaccharide; and nanoparticles comprising an amphiphilic non-ionizable block copolymer and the compound of Formula I of claim 1 or 2, wherein the pharmaceutical composition has a gel point of about 30°C to about 37°C.

11. The pharmaceutical composition according to any one of claims 3 to 10, wherein the pharmaceutical composition is for local administration.

12. The pharmaceutical composition according to any one of claims 3 to 11, wherein the pharmaceutical composition is for administration directly to the eye.

13. The pharmaceutical composition according to any one of claims 3 to 12, wherein the pharmaceutical composition is for administration directly to the back of the eye.

14. The pharmaceutical composition according to any one of claims 3 to 13, wherein the pharmaceutical composition is an intravitreal injection.

15. The pharmaceutical composition according to any one of claims 3 to 13, wherein the pharmaceutical composition is an eye drop pharmaceutical composition.

16. The pharmaceutical composition according to any one of claims 3 to claim 15, wherein the pharmaceutical composition is administered bimonthly, monthly, weekly, every other day, daily, twice per day, or three times per day.

17. A method of protecting a neuron from neurodegeneration or a method of treating neurodegeneration in a neuron, the method comprising contacting the neuron with the compound of claim 1 or 2 or the pharmaceutical composition according to any one of claims 3 to 16 in an amount (or concentration) sufficient to inhibit neurodegeneration in a subject in need thereof.

18. A method of treating age-related macular degeneration, the method comprising administering the compound according to claim 1 or 2 or the pharmaceutical composition according to any one of claims 3 to 16 to a subject in need thereof. -53- 1101228610\3\AMERICASAttorney Docket No.120754.00100 19. A method of improving retinal pigment epithelium integrity, comprising administering a compound according to claim 1 or 2 or the pharmaceutical composition according to any one of claims 3 to 16 to a subject in need thereof.

20. The method according to any one of claims 17 to 19, wherein the contacting or administering is to an eye of the subject in need thereof.

21. The method according to any one of claims 17 to 20, wherein the method is for protecting a neuron from neurodegeneration and the neuron is a retinal neuron.

22. The method according to any one of claims 17 to 21, wherein the method comprises simultaneously administering or contacting the neuron with the compound or pharmaceutical composition and an additional active agent for the treatment of age-related macular degeneration.

23. The method according to any one of claims 17 to 22, wherein the method comprises sequentially administering or contacting the neuron with the compound or pharmaceutical composition and an additional active agent for the treatment of age-related macular degeneration.

24. The method according to any one of claims 17 to 23, wherein the subject in need thereof is a mammal.

25. The method according to any one of claims 17 to 24, further comprising anaesthetizing the subject in need thereof prior to the contacting or administering.

26. The method according to any one of claims 17 to 25 further comprising dilating the pupil(s) of the subject in need thereof prior to the contacting or administering, optionally wherein for claim 25 the dilating step is before or concurrent with the anaesthetizing step.

27. The method according to any one of claims 17 to 26, wherein the method is for protecting a neuron from neurodegeneration and the neuron is light damaged.

28. The method according to any one of claims 17 to 27, wherein the amount administered is sufficient to inhibit neurodegeneration.

29. The method according to claim 28, wherein the amount (or concentration) sufficient to inhibit neurodegeneration is an amount (or concentration) sufficient to reduce the progression of age- related macular degeneration or vision loss associated therewith.

30. The method according to any one of claims 19 to 26, wherein the method is for improving retinal pigment epithelium integrity and / or reducing autofluorescence deposit. -54- 1101228610\3\AMERICASAttorney Docket No.120754.00100 31. The method according to claim 30, wherein an amount is administered and is sufficient for improving retinal pigment epithelium integrity and / or reducing autofluorescence deposit. -55- 1101228610\3\AMERICAS

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