Composition and method for the use of cannabinol compounds in neuroprotection

Abstract

Methods and compositions for neuroprotection are provided herein. Neuroprotective compositions may be 4-chlorobutyl-cannabinol (4-chlorobutyl-CBN) or its derivatives and / or pharmaceutically acceptable salts thereof, or may comprise them. Neuroprotective compositions may be used for the treatment of neurodegenerative diseases. Neuroprotective compositions may be used to protect retinal neurons from degeneration in subjects in need, such as for the treatment of age-related macular degeneration.

Description

Background Technology

[0001] Neurodegeneration is a phenomenon underlying various distinct diseases of the central and peripheral nervous systems. Neurodegeneration includes neuronal atrophy, axonal degeneration (e.g., Wallerian and / or Wallerian-like degeneration), and the induction of mechanisms of necrosis or programmed cell death. Different types of programmed cell death, such as apoptosis, autophagy, pyroptosis, and neoplasticity, 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 the mechanisms of neurodegeneration.

[0002] Ophthalmic diseases associated with neurodegeneration include age-related macular degeneration (AMD). Globally, AMD affects approximately 14 to 24% of the population aged 65 to 74 and about 35% of the population over 75, causing visual impairment or loss in the center of the visual field (macula) due to damage to the retina and / or associated neurons. AMD is a cause of vision loss and potential blindness in the population over the age of 50. The two main forms of AMD are atrophic (non-exudative or 'dry') AMD and neovascular (exudative or 'wet') AMD. Atrophic AMD is characterized by geographic atrophy (GA) in the center of the macula during the progression of AMD, and vision can gradually deteriorate over the years due to the loss of photoreceptors and the development of GA. Neovascular AMD is a more severe form of AMD characterized by neovascularization (e.g., choroidal neovascularization) in the progression of the disease, which can lead to rapid blindness. Neovascular AMD affects over 30 million patients worldwide and is a leading cause of vision loss in the population aged 60 and older. If AMD is left untreated, patients are likely to lose central vision in the affected eye within 24 months of onset. Approximately 90% of AMD patients have the dry form, while about 10% develop neovascular AMD.

[0003] Neuroprotection is an effect capable of providing relief or restoration of the nervous system, its cells, structures, and / or functions, or tolerance to neurodegenerative stimuli. Neuroprotective compositions may find applications in treating or alleviating the symptoms of various diseases that cause or lead to neurodegeneration, such as age-related macular degeneration. Despite significant progress in understanding the underlying mechanisms of neurodegeneration, there is still a need for methods and compositions for neuroprotection.

[0004] Cannabinoids, their salts, and their derivatives possess several properties that make them potentially therapeutic. Activation or blockade of CB1 and / or CB2 receptors using cannabinoids can modulate downstream signaling and metabolic pathways and, consequently, affect synaptic transmission, including the transmission of pain and other sensory signals, immune responses, and inflammation in the periphery. Therefore, there is interest in the use of natural or non-natural cannabinoids for therapeutic purposes. means of solving the problem

[0005] Compounds, pharmaceutical compositions, and methods of using the same are described herein. In one or more embodiments, the compound and the pharmaceutical composition are neuroprotective. The compound, or the pharmaceutical composition containing the compound, for example, the compound of Formula I, may come into contact with a neuron to provide a neuroprotective effect. In one or more embodiments, contact is made by administering the compound, for example, the compound of Formula I, or the pharmaceutical composition containing such a compound, for example, the compound of Formula I, to a subject requiring contact. The compound, for example, the compound of Formula I, the pharmaceutical composition containing such a compound, for example, the compound of Formula I, and the related method are useful for providing neuroprotection and, in some embodiments, for treating various ocular neurodegenerative diseases. In one or more embodiments, the compound, for example, the compound of Formula I, and the pharmaceutical composition containing such a compound, for example, the compound of Formula I, are provided for use in inducing a neuroprotective effect in retinal neurons, for example. For example, a compound, for example, a compound of Formula I, or a pharmaceutical composition comprising such a compound, for example, a compound of Formula I, may be administered topically or systemically to a subject to induce a neuroprotective effect in retinal neurons and may treat ocular neurodegenerative diseases such as, for example, AMD (including, for example, atrophic (non-exudative or 'dry') AMD and neovascular (exudative or 'wet') AMD).In some embodiments, the compound or pharmaceutical composition protects against or treats neurodegeneration in neurons, including neuronal atrophy, axonal degeneration (e.g., Waller and / or Waller-like degeneration) and / or the induction of mechanisms of necrosis or programmed cell death (e.g., programmed cell death such as apoptosis, autophagy, pyroptosis, and neoplasticity in neurons), 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 comprising the compound of Formula I are involved in the transmission of pain and other sensory signals in the periphery, immune responses, and inflammation. The compound of the first or second embodiment (and any embodiment thereof) or the pharmaceutical composition of the third embodiment (and any embodiment thereof) may be used to protect against or treat the pathological conditions of this paragraph.

[0006] In the first embodiment, a compound of the following formula I

[0007]

[0008] I

[0009] (Name: 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]cromen-1-ol or 4-chlorobutyl-cannabinol, or 4-chlorobutyl-CBN) or its derivatives, and / or their pharmaceutically acceptable salts are provided. 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.

[0010] In the second embodiment, a compound of the following formula I

[0011]

[0012] I

[0013] Or a pharmaceutically acceptable salt thereof is provided. In one or more embodiments, the compound of Formula I is not provided as a pharmaceutically acceptable salt.

[0014] In the third embodiment, a compound of the following formula I

[0015]

[0016] I

[0017] A pharmaceutical composition is provided comprising or its derivatives, and / or its pharmaceutically acceptable salts and pharmaceutically acceptable excipients. In one or more embodiments, a compound of Formula I according to the following formula is provided.

[0018]

[0019] I

[0020] A pharmaceutical composition comprising a compound of Formula I or a derivative thereof and / or a pharmaceutically acceptable excipient is provided. 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 only of ethanol. In some embodiments, the pharmaceutically acceptable excipient does not consist only of ethanol.

[0021] In a fourth embodiment, a method for protecting a neuron from neurodegeneration or treating neurodegeneration in a neuron is provided, the method comprising the step of contacting the neuron with a compound of the first or second embodiment (and any embodiment thereof) or a pharmaceutical composition of the third embodiment (and any embodiment thereof) in an amount (or concentration) sufficient to inhibit neurodegeneration in a subject requiring contact. In one or more embodiments, the contact is in vitro. In one or more embodiments, the contact is in vivo. In one or more embodiments, the contact comprises administering the compound or pharmaceutical composition to a subject requiring contact. In one or more embodiments, the neuron is a retinal neuron (e.g., a retinal ganglion). In one or more embodiments, the contact comprises administering the compound of the first or second embodiment (and any embodiment thereof) or the pharmaceutical composition of the third embodiment (and any embodiment thereof) as, for example, an intravitreal [IVT] injection. The intravitreal injection may be aimed at the back of the eye as illustrated 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]cromen-1-ol (i.e., not its derivatives or pharmaceutically acceptable salts). In one or more embodiments, the method is intended to protect a subject from AMD or to treat AMD in a subject who requires it. In some or any embodiments, the method is intended to protect a subject from glaucoma or to treat glaucoma in a subject. In some or any embodiments, the method is not intended to protect a subject from glaucoma and is not intended to treat glaucoma in a subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma.In some or any embodiment, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0022] In a fifth embodiment, a method for protecting a subject from neurodegeneration or treating neurodegeneration in a subject is provided, comprising the step of administering to a subject in need a compound of the first or second embodiment (and any embodiment thereof) or a pharmaceutical composition of the third embodiment (and any embodiment thereof). In one or more embodiments, the pharmaceutical composition is 4-chlorobutyl-CBN, or a pharmaceutically acceptable salt thereof, or a derivative thereof (e.g., ,It is suitable for achieving a neuroprotective dose of a prodrug. In one or more embodiments, the compound of the first or second embodiment (and any of its embodiments) or the pharmaceutical composition of the third embodiment (and any of its embodiments) is formulated for administration to the eye and is intended for use in protecting neurons from neurodegeneration or treating neurodegeneration in neurons. In one or more embodiments, the compound of the first or second embodiment (and any of its embodiments) or the pharmaceutical composition of the third embodiment (and any of its embodiments) is formulated to achieve a concentration of about 0.15 μM to less than about 20 μM of 4-chlorobutyl-CBN, or its pharmaceutically acceptable salt, or its derivative (e.g., a prodrug) in the vitreous humor, in ocular tissues of the eye, and / or in contact with ocular neuron cells. In one or more embodiments, the ocular neuron cells are retinal neurons or other neuron cells (e.g., including but not limited to retinal ganglia and photoreceptors). In one or more embodiments, the method is intended to protect a subject from AMD or to treat AMD in a subject who requires it. In some or any embodiments, the method is intended to protect a subject from glaucoma or to treat glaucoma in a subject. In some or any embodiments, the method is not intended to protect a subject from glaucoma and is not intended to treat glaucoma in a subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma. In some or any embodiments, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0023] In the sixth embodiment, preferably according to one or more of the aforementioned embodiments (fourth and fifth embodiments) and any of the embodiments provided herein, use of the compound of the first or second embodiment (and any of the embodiments thereof) or the pharmaceutical composition of the third embodiment (and any of the embodiments thereof) for treating neurodegeneration in a subject in need is provided by using the pharmaceutical composition described herein or according to the method described herein. In one or more embodiments, the method is intended to protect the subject from AMD or to treat AMD in a subject in need. In some or any embodiments, the method is intended to protect the subject from glaucoma or to treat glaucoma in the subject. In some or any embodiments, the method is not intended to protect the subject from glaucoma and is not intended to treat glaucoma in the subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma. In some or any embodiments, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0024] In a seventh embodiment, a method for treating age-related macular degeneration is provided, comprising the step of administering a compound of the first or second embodiment (and any embodiment thereof) or a pharmaceutical composition of the third embodiment (and any embodiment thereof) to a subject in need.

[0025] In an eighth embodiment, a method for improving RPE integrity in an AMD subject and reducing autofluorescent deposits characteristic of dry AMD is provided, comprising the step of administering a compound of the first or second embodiment (and any embodiment thereof) or a pharmaceutical composition of the third embodiment (and any embodiment thereof) to a subject in need. In one or more embodiments, a method is provided in which an amount sufficient to improve retinal pigment epithelial integrity is administered.

[0026] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth embodiments (and any embodiments of the embodiments), contact and / or administration involves administering topically to a subject in need. For example, administration is made to a subject suffering from neurodegeneration, e.g., neurodegeneration of the eye. In one or more embodiments, contact and / or administration involves administering to a subject suffering from a neurodegenerative disease, e.g., neurodegenerative disease of the eye. In one or more embodiments, contact and / or administration involves administering to a subject suffering from AMD. In one or more embodiments, contact and / or administration involves administering to a subject diagnosed with AMD. In one or more embodiments, the method comprises the step of contacting and / or administering an additional activator for the treatment of AMD to a subject in need, either simultaneously or sequentially. In some or any embodiments, the subject suffers from glaucoma. In some or any embodiments, the subject does not suffer from glaucoma.

[0027] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth embodiments (and any embodiments of the embodiments), the amount is an amount (or concentration) sufficient to reduce the amount or rate of apoptosis of a population of neurons in contact with the compound or pharmaceutical composition. In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration is an amount (or concentration) sufficient to reduce the progression of age-related macular degeneration or associated vision loss. 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]cromen-1-ol (i.e., not its derivatives or pharmaceutically acceptable salts). In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect retinal neurons, protect retinal tissue, protect photoreceptor cells, protect against or reduce the accumulation of autofluorescent extracellular debris, and protect against or treat neuroinflammation. In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect against or treat neuroinflammation. In some or any embodiments, the subject has glaucoma. In some or any embodiments, the subject does not have glaucoma.

[0028] In one or more embodiments of any of the 4th, 5th, 6th, 7th and 8th embodiments (and any embodiment of the embodiments), an amount (or concentration) sufficient to inhibit neurodegeneration or sufficient to improve retinal pigment epithelial integrity is about 0.15 μM to less than about 20 μM of 4-chlorobutyl-CBN (or its pharmaceutically acceptable salts or derivatives (e.g., prodrugs), in some embodiments, wherein the amount is the amount of 4-chlorobutyl-CBN other than the salts and prodrugs) in the vitreous humor, in the ocular tissues of the eye and / or in contact with ocular neuronal cells. In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration or an amount (or concentration) sufficient to improve retinal pigment epithelial integrity is an amount that results in a concentration selected from about 0.3 μM to about 15 μM or less of 4-chlorobutyl-CBN in the vitreous humor, in the ocular tissue of the eye and / or in contact with ocular neuronal cells. In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration or an amount (or concentration) sufficient to improve retinal pigment epithelial integrity is an amount that results in a concentration of 4-chlorobutyl-CBN of about 0.5 μM or more and about 15 μM or less in the vitreous humor, in the ocular tissue of the eye and / or in contact with ocular cells, e.g., 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 selected from. 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]cromen-1-ol (i.e., not its derivatives or pharmaceutically acceptable salts).In one or more embodiments, the amount is based on the compound excluding any pharmaceutically acceptable salt. In some or any embodiments, the subject has glaucoma. In some or any embodiments, the subject does not have glaucoma.

[0029] In one or more embodiments of any of the 4th, 5th, 6th, 7th, and 8th embodiments (and any embodiments of the embodiments), an amount (or concentration) sufficient to inhibit neurodegeneration or an amount (or concentration) sufficient to improve retinal pigment epithelial integrity is in the vitreous fluid, in ocular tissues of the eye, and / or in contact with ocular cells of 4-chlorobutyl-CBN, its pharmaceutically acceptable salts, or its derivatives (e.g., prodrugs). It is an amount resulting in a concentration selected from about 0.15 μM to less than about 20 μM, e.g., selected from about 0.5 μM to less than about 15 μM, selected from more than about 2 μM to less than about 12 μM, 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. In one or more embodiments, the subject has AMD. In some or any embodiments, the subject has glaucoma. In some or any embodiments, the subject does not have glaucoma.

[0030] In one or more embodiments, an amount (or concentration) sufficient to inhibit neurodegeneration or an amount (or concentration) sufficient to improve retinal pigment epithelial integrity is an amount that results in a concentration range having a lower limit of a range selected from any one of 0.15, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, and 9 μM and an upper limit of a 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, wherein the lower limit and the upper limit form a mathematically acceptable range, and the concentration is in contact with 4-chlorobutyl-CBN, its pharmaceutically acceptable salt or its derivative (e.g., in the vitreous humor, in ocular tissues of the eye, and / or ocular cells) It is the concentration of the prodrug. In one or more embodiments, the subject has AMD. In some or any embodiments, the subject has glaucoma. In some or any embodiments, the subject does not have glaucoma.

[0031] Fourth, fifth, sixth, seventh, and eighth modes In one or more embodiments of any of the embodiments (and any embodiments of the embodiments), the pharmaceutical composition has a final concentration (in vivo) of the compound of Formula I that is about 1 / 10 (1 / 10) of the initial concentration of the compound of Formula I. In one or more embodiments, the pharmaceutical composition has a final concentration (in vivo) of the compound of Formula I that is 1 / 10 of the initial concentration of the compound of Formula I. In one or more embodiments, the initial concentration is the concentration of 4-chlorobutyl-CBN (or its pharmaceutically acceptable salt or its derivative) in the pharmaceutical composition. In one or more embodiments, if the pharmaceutical composition is an intravitreal injection, the initial concentration is the concentration of 4-chlorobutyl-CBN (or its pharmaceutically acceptable salt or its derivative) in the intravitreal injection. In one or more embodiments, the final concentration is the concentration of 4-chlorobutyl-CBN (or its pharmaceutically acceptable salt or its derivative) in the vitreous fluid, in the ocular tissue 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 its pharmaceutically acceptable salt or its derivative) in the vitreous fluid. 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]cromen-1-ol (i.e., not its derivative or pharmaceutically acceptable salt). In one or more embodiments, the concentration is based on the compound excluding any pharmaceutically acceptable salt.

[0032] In one or more embodiments, 4-chlorobutyl-CBN is provided in a sustained-release formulation. In one or more embodiments, the formulation comprises a) a delivery carrier comprising a cellulose polymer and an anionic polysaccharide, and b) nanoparticles comprising an amphiphilic non-ionic block copolymer and 4-chlorobutyl-CBN, wherein the formulation has a gelation point of about 30 °C to about 37 °C.

[0033] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth embodiments (and any embodiments of the embodiments), contact and / or administration comprises systemic administration. In one or more embodiments, systemic administration comprises intravenous injection. In one or more embodiments, systemic administration comprises oral administration. In one or more embodiments, systemic administration comprises transdermal administration. In one or more embodiments, 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]cromen-1-ol (i.e., not its derivatives or pharmaceutically acceptable salts). In one or more embodiments, the method is intended to protect a subject from AMD or to treat AMD in a subject who requires it. In some or any of the embodiments, the method is intended to protect a subject from glaucoma or to treat glaucoma in a subject. In some or any embodiments, the method is not intended to protect the subject from glaucoma and is not intended to treat glaucoma in the subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma. In some or any embodiments, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0034] In one or more embodiments of any of the fourth, fifth, sixth, seventh, and eighth embodiments (and any embodiments of the embodiments), contact and / or administration involves local administration. In one or more embodiments, contact and / or administration is made directly to the eye. For example, contact and / or administration is made on the eye (e.g., as eye drops, such as in the form of microemulsion eye drops, or as an eye gel). As another example, contact and / or administration is made directly into or behind the eye (e.g., via intravitreal injection or a pump). In one or more embodiments, the method is intended to protect a subject from AMD or to treat AMD in a subject who requires it. In some or any embodiments, the method is intended to protect a subject from glaucoma or to treat glaucoma in a subject. In some or any embodiments, the method is not intended to protect a subject from glaucoma and is not intended to treat glaucoma in a subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma. In some or any embodiment, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0035] Inclusion by reference

[0036] All publications, patents, and patent applications mentioned in this specification are incorporated by reference into this specification to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being included by reference. Brief explanation of the drawing

[0037] FIG. 1 illustrates a half-sectional view of the eye, showing parts of the eye and an intravitreal injection site for the treatment of age-related macular degeneration according to one or more embodiments of the present invention. Figure 2a illustrates the results of a flash electroretinogram (fERG) test, showing the response of photoreceptors in the eye to light stimulation (luminance) with an intensity (luminance) of 0.01 cd*s / m^2. The α-wave amplitude (μV) represents the response of the photoreceptors, and the luminance (cd*s / m^2) represents the intensity of the flash of light. Control group: Healthy animals without light damage (LD) and without any treatment. Vehicle: Animals with light damage treated with sterile diluted buffer. Figure 2b illustrates the results of scintillation electroretinography (fERG) testing, showing the response of photoreceptors in the eye to light stimulation (luminance) of an intensity of 0.01 cd*s / m^2. Normalized α-wave amplitude % is plotted for healthy animals (CTRL or control = 100%). Control: Healthy animals without photoinjury and receiving no treatment. Vehicle: Animals with photoinjury and treated with dilution buffer. Figure 3a illustrates the results of scintillation electroretinography (fERG), showing the response of photoreceptors in the eye to light stimulation at increasing intensity levels. α-wave amplitude (μV) represents the photoreceptor response, and luminance (cd*s / m^2) represents the intensity of the light scintillation. In the eyes injected with 4-chlorobutyl-CBN, an increase in α-wave amplitude was observed for individual light stimuli (measured in luminance) compared to the vehicle. In the eyes injected with CBN, no overall improvement in α-wave amplitude was observed compared to the vehicle. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle: Animals with photodamage and treated with dilution buffer. Figure 3b illustrates the alpha-wave amplitude at an intensity of 0.01 cd*s / m^2 (from Figure 3a) to better illustrate the increase in alpha-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot alpha-wave amplitude (μV) (left) and normalized alpha-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 3c illustrates the alpha-wave amplitude at an intensity of 0.1 cd*s / m^2 (from Figure 3a) to better illustrate the increase in alpha-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot alpha-wave amplitude (μV) (left) and normalized alpha-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 3d illustrates the alpha-wave amplitude at an intensity of 1 cd*s / m^2 (from Figure 3a) to better illustrate the increase in alpha-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot alpha-wave amplitude (μV) (left) and normalized alpha-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 3e illustrates the alpha-wave amplitude at an intensity of 10 cd*s / m^2 (from Figure 3a) to better illustrate the increase in alpha-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot alpha-wave amplitude (μV) (left) and normalized alpha-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 3f illustrates the alpha-wave amplitude at an intensity of 100 cd*s / m^2 (from Figure 3a) to better illustrate the increase in alpha-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot alpha-wave amplitude (μV) (left) and normalized alpha-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 4a illustrates the results of scintillation electroretinography (fERG), showing the response of the inner retina to light stimulation at increasing intensity levels. b-wave amplitude (μV) represents the response of the inner retina, and luminance (cd*s / m^2) represents the intensity of the light scintillation. In the eyes injected with 4-chlorobutyl-CBN, an increase in b-wave amplitude was observed for individual light stimulation (measured in luminance) compared to the vehicle. In the eyes injected with CBN, no overall improvement in b-wave amplitude was observed compared to the vehicle. Control group: Healthy animals without photodamage and without treatment. Vehicle: Animals with photodamage treated with dilution buffer. Figure 4b illustrates the b-wave amplitude at an intensity of 0.1 cd*s / m^2 (from Figure 4a) to better illustrate the increase in b-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot b-wave amplitude (μV) (left) and normalized b-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 4c illustrates the b-wave amplitude at an intensity of 1 cd*s / m^2 (from Figure 4a) to better illustrate the increase in b-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms show b-wave amplitude (μV) (left) and normalized b-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 4d illustrates the b-wave amplitude at an intensity of 10 cd*s / m^2 (from Figure 4a) to better illustrate the increase in b-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot b-wave amplitude (μV) (left) and normalized b-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 4e illustrates the b-wave amplitude at an intensity of 100 cd*s / m^2 (from Figure 4a) to better illustrate the increase in b-wave amplitude occurring upon 4-chlorobutyl-CBN treatment. Histograms plot b-wave amplitude (μV) (left) and normalized b-wave amplitude (%) for healthy animals (CTRL or control = 100%) (right). Control: Healthy animals without photodamage and receiving no treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 5 illustrates the results of scintillation electroretinography (fERG) tests, showing the response of the eye's inner retinal circuit to light stimulation (primarily caused by rod bipolar and A11 / A17 amacrine cells) at increasing intensity levels. Oscillatory potential (OP) amplitude (μV) represents the response of the inner retina, and luminance (cd*s / m^2) represents the intensity of the scintillation of light. In the eyes injected with 4-chlorobutyl-CBN, an increase in OP amplitude was observed for individual light stimulation (measured in luminance) compared to the vehicle. In the eyes injected with CBN (30 μM), an increase in OP amplitude was observed at 100 cd*s / m^2 (dark adaptation stimulation) compared to the vehicle, whereas no overall improvement in OP amplitude was observed for other light stimulations compared to the vehicle. In eyes injected with CBN (100 μM), no overall improvement in OP amplitude was observed compared to the vehicle for any applied photostimulus. Control group: Healthy animals with no photodamage and no treatment. Vehicle: Animals with photodamage and treated with dilution buffer. Figure 6a illustrates in vivo imaging of the inner retinal layer structure of a photodamaged rat model. The imaging shows the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL) within cells treated with 4-chlorobutyl-CBN (3 μM). Control: Healthy animals without photodamage and without treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 6b illustrates in vivo imaging of the inverse (negative) exposure of the inner retinal layer structure. The imaging shows the outer nuclear layer, inner nuclear layer, and ganglion cell layer within cells treated with 4-chlorobutyl-CBN (3 μM). Control: Healthy animals without photodamage and without treatment. Vehicle: Photodamaged and treated with dilution buffer. Figure 7a illustrates the alpha-wave amplitudes in response to different dose levels (intra-eye) of 0.3 μM, 1 μM, 3 μM, and 10 μM of 4-Cl-butyl-CBN. An overall increase in alpha-wave response is observed in eyes treated with 3 μM and 10 μM of 4-Cl-butyl-CBN compared to the vehicle. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle group: Animals with photodamage and treated with dilution buffer. Figure 7b illustrates the b-wave amplitudes in response to different dose levels (intra-eye) of 0.3 μM, 1 μM, 3 μM, and 10 μM of 4-Cl-butyl-CBN. An overall increase in the b-wave response is observed in eyes treated with 3 μM and 10 μM of 4-Cl-butyl-CBN compared to the vehicle. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle group: Animals with photodamage and treated with dilution buffer. Figure 7c illustrates the oscillatory potential (OP) amplitudes in response to different dose levels (intra-eye) of 0.3 μM, 1 μM, 3 μM, and 10 μM of 4-Cl-butyl-CBN. An overall increase in OP response is observed in eyes treated with 3 μM and 10 μM of 4-Cl-butyl-CBN compared to the vehicle. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle group: Animals with photodamage and treated with dilution buffer. Fig. 8a illustrates a histological imaging of the retina for measuring the thickness of the outer nuclear layer (ONL). Scale bar: 200 μm. Figure 8b illustrates the ratio of ONL to retinal thickness at various locations along the superior and inferior retinas. Data are plotted as mean + / - SE. Statistical analysis was performed on the Student's between the vehicle and 4-Cl-butyl-CBN (3 μM). t - Tested (n=7 / group). Compared to vehicle, *p<0.05, ***p<0.001. Compared to vehicle, ONL: an increase in retinal thickness is observed for 4-Cl-butyl-CBN. FIG. 8c illustrates a schematic diagram of a retinal cryosection showing the retinal locations shown in FIG. 8b along the superior and inferior retinas. ON = optic nerve. Figure 9a illustrates the histological imaging of autofluorescence (AF) extracellular deposits. A reduction in extracellular deposits for 4-Cl-butyl-CBN is observed compared to the vehicle. Figure 9b illustrates the number of AF deposits. A statistically significant reduction in extracellular deposits for 4-Cl-butyl-CBN is observed compared to the vehicle. Data are plotted as mean + / - SE. Statistical analysis of the Student between the vehicle and 4-Cl-butyl-CBN (3 μM) t - It was a test (n=7 / group). Vehicle comparison **p<0.01. Figure 10a illustrates histological imaging of the retinal pigment epithelium (RPE) using anti-RPE65. Improvement in RPE integrity is observed for 4-Cl-butyl-CBN compared to the vehicle. Figure 10b illustrates the normalized RPE length for the control group. A statistically significant increase in RPE length was observed between the 4-Cl-butyl-CBN and the control group. Data are plotted as mean + / - SE. Statistical analysis was performed on the Student between the vehicle and 4-Cl-butyl-CBN (3 μM). t - It was a test (n=7 / group). Compared to vehicle ***p<0.001. These data show that 4-Cl-butyl-CBN improved RPE length in the AMD model. Figure 11a illustrates the immunostaining of a retinal cross-section 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. Figure 11b illustrates the number of IBA-1 (+) cells in the outer retina (subretinal, outer nucleus, outer reticular layer) from the upper to the lower margin of the retina. A reduction in retinal inflammation is observed for 4-Cl-butyl-CBN compared to vehicle data. Data are plotted as mean + / - SE. Statistical analysis of the Student's t - It was a test (n=7 / group). Vehicle comparison *p<0.05, **p<0.01. Figure 11c illustrates the number of IBA-1 (+) cells in the epiretina. A statistically significant reduction in inflammation in the epiretina is observed for 4-Cl-butyl-CBN compared to the vehicle. Data are plotted as mean + / - SE. Statistical analysis of the Student's t - It was a test (n=7 / group). Vehicle comparison *p<0.05, **p<0.01. Figure 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 the vehicle. Data are plotted as mean + / - SE. Statistical analysis of the Student's t - It was a test (n=7 / group). Vehicle comparison *p<0.05, **p<0.01. Figure 11e illustrates the number of IBA-1 (+) cells in the superior and inferior retinas. A statistically significant reduction in inflammation in the retina is observed for 4-Cl-butyl-CBN compared to the vehicle. Data are plotted as mean + / - SE. Statistical analysis of the Student's t- It was a test (n=7 / group). Vehicle comparison *p<0.05, **p<0.01. Figure 12 illustrates the concentrations of 4-Cl-butyl-CBN administered topically to rabbits or IVT to rats. Increased concentrations are observed in the retina / RPE / choroid for IVT administration to rats compared to topical application to rabbits. Specific details for implementing the invention

[0038] A compound, e.g., a compound of Formula I, and a pharmaceutical composition thereof, and a method of such compound, e.g., a compound of Formula I, and a pharmaceutical composition thereof for protecting neurons from one or more cytotoxic stimuli are described herein. In one or more embodiments, the method comprises the step of bringing a neuron into contact with the compound, e.g., a compound of Formula I, or the pharmaceutical composition thereof, by administering the compound or pharmaceutical composition to a subject in need, for example. The method, the compound, e.g., a compound of Formula I, and the pharmaceutical composition thereof described herein find specific uses for the protection of retinal neurons, but are not limited thereto. In some cases, the method, the compound, e.g., a compound of Formula I, and the pharmaceutical composition thereof described herein may be used for the neuroprotection of retinal neurons in a subject, e.g., a subject suffering from age-related macular degeneration. In this way, the method, the compound, e.g., a compound of Formula I, and the pharmaceutical composition thereof may be described as neuroprotective. In one or more embodiments, the neuron is a damaged neuron, including but not limited to photodamaged neurons. In one or more embodiments, the compound, for example, a compound of Formula I, and the pharmaceutical composition thereof comprises 4-chlorobutyl-CBN. In some cases, the method comprises the step of bringing retinal neurons into contact with 4-chlorobutyl-CBN, for example, by administering 4-chlorobutyl-CBN (or the pharmaceutical composition thereof) to a subject in need. In one or more embodiments, the method is intended to protect the subject from AMD or to treat AMD in a subject in need. In some or any embodiments, the method is intended to protect the subject from glaucoma or to treat glaucoma in the subject. In some or any embodiments, the method is not intended to protect the subject from glaucoma and is not intended to treat glaucoma in the subject. In some or any embodiments, the method is intended to reduce intraocular pressure in a subject suffering from glaucoma.In some or any embodiment, the method is not intended to reduce intraocular pressure in a subject suffering from glaucoma.

[0039] In one or more embodiments, the compound is 4-chlorobutyl-CBN having the chemical formula of the following chemical formula I.

[0040]

[0041] I

[0042] In one or more embodiments, the compound is a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0043] The pharmaceutical composition may be 4-chlorobutyl-CBN having the chemical formula I below, or may contain the same.

[0044]

[0045] I

[0046] In one or more embodiments, the pharmaceutical composition 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.

[0047] In one or more embodiments, the compound or pharmaceutical composition may be or contain a prodrug or a pharmaceutically acceptable salt of the compound of Formula I. For example, the compound or pharmaceutical composition may be or contain a prodrug of 4-chlorobutyl-CBN or a pharmaceutically acceptable salt thereof.

[0048] In some cases, the prodrug is an ester of 4-chlorobutyl-CBN (according to the chemical 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. Further prodrug strategies for the compounds described herein are found in U.S. Patent Publications Nos. 2016 / 0228490, 2011 / 0052694, 2015 / 0197484, 2008 / 0076789, 2009 / 0143462, 2012 / 0289484, 2009 / 0036523, 2009 / 0156814 and 2008 / 0008745, and the literature [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 incorporated herein for all purposes and in particular the 4-chlorobutyl-CBN prodrug and pharmaceutical compositions containing the same, and methods of using and / or administering the prodrug as described herein.

[0049] As used herein, the term 'prodrug' refers to a derivative that is a precursor compound that releases a biologically active compound in vivo through certain chemical or physiological processes after administration (e.g., reaching a physiological pH or conversion of the prodrug into a biologically active compound via enzymatic action). The prodrug itself may lack or possess the desired biological activity. Therefore, the term 'prodrug' refers to a precursor of a pharmaceutically acceptable biologically active compound. In certain cases, the prodrug possesses improved physical and / or delivery properties compared to the parent compound from which it is derived. Prodrugs often offer advantages in solubility, histocompatibility, or delayed release in mammalian organisms (Reference [H. Bundgard, Design of Prodrugs (Elsevier, Amsterdam, 1988), pp. 7-9, 21-24]). Discussion of prodrugs is [T. Higuchi et al., "Pro-Drugs as Novel Delivery Systems," ACS Symposium Series, Vol.

[14] and provided in [EB Roche, ed., Bioreversible Carriers in Drug Design (American Pharmaceutical Association & Pergamon Press, 1987)]. Exemplary benefits of prodrugs may include, but are not limited to, their physical properties such as enhanced drug stability for long-term storage.

[0050] The term “prodrug” also means comprising any covalently bonded carrier that releases an active compound in vivo when the prodrug is administered to a subject. A prodrug of a therapeutically active compound as described herein may be prepared by modifying one or more functional groups present in a therapeutically active compound, namely a compound of Formula I, or a pharmaceutically acceptable salt thereof, and other therapeutically active compounds used in a method according to one or more embodiments or included in a pharmaceutical composition according to one or more embodiments, in such a manner that the modification is cleaved in a conventional operation or in vivo to produce a parent therapeutically active compound. The prodrug comprises a compound of Formula I, wherein the hydroxyl group is covalently bonded to any group that is cleaved to form a free hydroxyl group when the prodrug of the active compound is administered to a subject. Examples of prodrugs include, but are not limited to, formate or benzoate derivatives of alcohols available for reaction.

[0051] In some cases, the prodrug is 3,6,9,12-tetraoxatridecanoyl ester, N,N-dimethylglycyl ester, 3,6,9,12-tetraoxatridecyl carbonate, N-formylglycyl ester, N-formylsarcosyl ester, 3,6,9,12-tetraoxatridecyl oxalate, hemisuccinate, 4-aminobutyl carbamate, prolyl ester, 3-dimethylaminopropionate, glycolate, (D)-ribonate, ammonium phosphate, (R)-2,3-dihydroxypropyl carbonate, 3-hydroxy-2-(hydroxymethyl)-2-methylpropanoate, glycinate, β-alanineate, (S)-2,3-dihydroxypropanoate, (S)-2,3-dihydroxypropyl carbonate or It contains (R)-2,3-dihydroxypropyl carbonate.

[0052] Similarly, where the disclosed compound or a pharmaceutically acceptable form of the compound contains an alcohol functional group, the prodrug may be formed by replacing a 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, α-amino(C1-C4)alkanoyl, aryl acyl and α-aminoacyl, or α-aminoacyl-α-aminoacyl, wherein each α-aminoacyl group is a naturally occurring L-amino acid, P(O)(OH)2, P(O)(O(C1-C6)alkyl)2, or glycosyl (a hemiacetal form of a carbohydrate). It is independently selected from radicals resulting from the removal of hydroxyl groups.

[0053] The use of prodrug systems is discussed in the literature [T. This is described in et al., "Design and Pharmaceutical Applications of Prodrugs" in Drug Discovery Handbook (SC Gad, ed., Wiley-Interscience, Hoboken, NJ, 2005), ch. 17, pp. 733-796). Other alternatives for the composition and use of prodrugs are known in the art. Where a method or pharmaceutical composition according to one or more embodiments uses or contains a prodrug of cannabinol or 4-chlorobutyl-CBN, or other therapeutically active preparations, the prodrug and active metabolite of the compound may be identified using techniques known in the art. For example, in the literature [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).

[0054] Exemplary prodrugs useful in one or more embodiments of the present disclosure include, but are not limited to, a prodrug of a compound of Formula I below or a pharmaceutically acceptable salt thereof, and

[0055]

[0056] Here, X and Y may be the same or different and are 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., quaternary 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, hydravamin, isopropylamine, methylglucamine, morpholine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, tetramethylammonium hydroxide, benzyltrimethylammonium hydroxide, tris(hydroxymethyl)aminomethane (TRIS), N-(2-hydroxyethyl)pyrrolidine, piperazine, glucosamine, arginine, lysine, and histidine. In one or more embodiments, X and Y are different substituents. In one or more embodiments, X and Y are the same substituent. In one or more embodiments, X and Y may both be part of the same functional group, such as piperazine. In one or more embodiments, the phosphate is selected from the group consisting of diphosphate and triphosphate.

[0057] In one or more embodiments, the prodrug is a prodrug of a compound of the following formula I or a pharmaceutically acceptable salt thereof, and

[0058]

[0059] Here, R 4 is alkyl, alkoxyalkyl, alkylamino, hydroxyalkyl, or hydroxyalkylamino, and in one or more embodiments R 4 comprises 1 to 12 carbons and optionally 4 or fewer substitutions, and in one or more embodiments R 4 It includes 1 to 6 carbons and optionally 2 or fewer substitutions.

[0060] In one or more embodiments, the prodrug is a prodrug of a compound of the following formula I or a pharmaceutically acceptable salt thereof, and

[0061]

[0062] Here, R 4 is alkyl, alkoxyalkyl, alkylamino, hydroxyalkyl, or hydroxyalkylamino, and in one or more embodiments R 4 comprises 1 to 12 carbons and optionally 4 or fewer substitutions, and in one or more embodiments R 4 It comprises 1 to 6 carbons and optionally 2 or fewer substitutions. .

[0063] In one or more embodiments, the prodrug is a prodrug of a compound of the following formula I or a pharmaceutically acceptable salt thereof, and

[0064]

[0065] Here, R 4 is alkyl, alkoxyalkyl, alkylamino, hydroxyalkyl, or hydroxyalkylamino, and in one or more embodiments R 4 comprises 1 to 12 carbons and optionally 4 or fewer substitutions, and in one or more embodiments R 4 It comprises 1 to 6 carbons and optionally 2 or fewer substitutions. .

[0066] In one or more embodiments, the prodrug is a prodrug of a compound of the following formula I or a pharmaceutically acceptable salt thereof, and

[0067]

[0068] Here, R 4 is alkyl, alkoxyalkyl, alkylamino, hydroxyalkyl, or hydroxyalkylamino, and in one or more embodiments R 4 comprises 1 to 12 carbons and optionally 4 or fewer substitutions, and in one or more embodiments R 4 It includes 1 to 6 carbons and optionally 2 or fewer substitutions.

[0069] In one or more embodiments of the present disclosure, a useful prodrug comprises, but is not limited to, a prodrug of a compound of Formula I below or a pharmaceutically acceptable salt thereof.

[0070]

[0071] In one or more embodiments of the present disclosure, a useful prodrug comprises, but is not limited to, a prodrug of a compound of Formula I below or a pharmaceutically acceptable salt thereof.

[0072] .

[0073] In one or more embodiments of the present disclosure, a useful prodrug comprises, but is not limited to, a prodrug of a compound of Formula I below or a pharmaceutically acceptable salt thereof.

[0074] or

[0075] In one or more embodiments, the aforementioned prodrug may be advantageously formulated into a cyclodextrin such as randomly methylated beta-cyclodextrin, 2-hydroxypropyl beta-cyclodextrin, or sulfobutyl ether beta-cyclodextrin.

[0076] The pharmaceutical composition may contain additional active agents. In one or more embodiments, the pharmaceutical composition may contain 4-chlorobutyl-CBN, its pharmaceutically acceptable salt, or its derivative, and additional cannabinoids or terpenoids. In one or more embodiments, the pharmaceutical composition may contain additional active pharmaceutical agents for the treatment of age-related macular degeneration.

[0077] Currently, different classes of treatments, including but not limited to micronutrient supplements, vascular-endothelial derived growth factor (VEGF)-A antagonists (anti-VEGF agents or VEGF inhibitors), and photodynamic therapy, are used to treat age-related macular degeneration.

[0078] 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, pariximab, and bevacizumab. Useful photodynamic therapy agents include, but are not limited to, verteporfin. In one or more embodiments, different therapeutic agents may be combined.

[0079] In one or more embodiments, the compounds and pharmaceutical compositions described herein containing, for example, 4-chlorobutyl-CBN or its pharmaceutically acceptable salts or derivatives, enable lower doses or less frequent administration of one or more therapeutic agents for the treatment of age-related macular degeneration.

[0080] definition

[0081] When referring to the compounds provided herein, unless otherwise specified, the following terms have the following meanings. Unless otherwise defined, all technical and scientific terms used herein may be used in the sense that is commonly understood by those skilled in the art. Where there are multiple definitions of a term in this section, the definitions in this section shall prevail unless otherwise noted. Unless otherwise specified, where a term is defined as being substituted, the groups in the list of substituents are themselves not substituted.

[0082] In this document, references to a value or parameter as “about” include (and describe) variations directed toward that value or parameter itself. For example, a description referring to “about X” includes a description of “X”. As used herein and unless otherwise specified, the terms “about” and “about”, when used in relation to the temperature, volume, amount, or weight percentage of a component in the form of a composition or dosage, mean a volume, amount, or weight percentage recognized by a person skilled in the art to provide a pharmaceutical effect equivalent to that obtained from a specified volume, amount, or weight percentage. Specifically, when used in this context, the terms “about” and “about” consider a volume, amount, or weight percentage within 15%, 10%, 5%, 4%, 3%, 2%, 1%, or 0.5% of a specified volume, amount, or weight percentage.

[0083] As used herein, the terms ‘one (a)’ or ‘some (an)’ mean one or more unless the context clearly indicates otherwise. For example, ‘pharmaceuticalally acceptable excipients’ refers to ‘one or more pharmaceutically acceptable excipients’.

[0084] As used herein, 'alkyl' refers to a linear or branched saturated monovalent hydrocarbon radical having 1 to 6 carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl, butyl, pentyl, etc. In one or more embodiments, the alkyl is a (C1-C6)alkyl.

[0085] As used herein, 'alkoxy' refers to a -OR group in which R is alkyl as defined herein. In one or more embodiments, the alkoxy is a (C1-C6)alkoxy.

[0086] As used herein, '(C1-C6)alkanoylyoxymethyl' refers to -CH2OC(O)R in which R is (C1-C6)alkyl as defined herein.

[0087] As used herein, '1-((C1-C6))alkanoyloxy)ethyl' is defined as R is (C1-C6)alkyl It refers to.

[0088] As used herein, '1-methyl-1-((C1-C6)alkanoyloxy)ethyl' is defined herein as R is a (C1-C6)alkyl It refers to.

[0089] As used herein, '(C1-C6)alkoxycarbonyloxymethyl' refers to -CH2OC(O)R in which R is (C1-C6)alkoxy as defined herein.

[0090] As used herein, 'N(C1-C6)alkoxycarbonylaminomethyl' refers to -CH2NHC(O)R in which R is (C1-C6)alkoxy as defined herein.

[0091] As used herein, 'succinoyl' refers to -C(O)CH2CH2C(O)OH.

[0092] As used herein, '(C1-C6)alkanoyle' refers to -C(O)R in which R is (C1-C6)alkyl as defined herein.

[0093] As used herein, 'α-amino(C1-C4)alkanoyl' refers to -C(O)R, an alkyl group in which R is substituted with -NH2.

[0094] As used herein, 'arylacyl' refers to -C(O)phenyl or -C(O)naphthyl.

[0095] As used herein, 'alkoxyalkyl' refers to an alkyl group as defined herein, substituted with one or two alkoxy groups as defined herein.

[0096] As used herein, 'alkylamino' refers to an alkyl group as defined herein, substituted with one or two -NH2 groups.

[0097] As used herein, 'hydroxyalkyl' refers to an alkyl group as defined herein that is substituted with one or two hydroxyl groups.

[0098] As used herein, 'hydroxyalkylamino' refers to -NHR, where R is a hydroxyalkyl as defined herein.

[0099] As used herein, 'objects requiring' etc. refer to mammals, preferably humans.

[0100] As used herein, '4-chlorobutyl-cannabinol', '4-chlorobutyl-CBN', '4-Cl-butyl-CBN', or '4-Cl-CBN' refers to 3-(4-chlorobutyl)-6,6,9-trimethyl-6H-benzo[c]cromen-1-ol.

[0101] “Salt” refers to an acid or base salt of a compound used in one or more methods of the present disclosure. Exemplary examples of pharmaceutically acceptable salts are salts of inorganic acids (hydrochloric acid, hydrobromide, phosphoric acid, etc.), salts of organic acids (acetic acid, propionic acid, glutamic acid, citric acid, etc.), and salts of quaternary ammoniums (methyl iodide, ethyl iodide, etc.). Pharmaceutically acceptable salts are understood to be non-toxic. Further information on suitable pharmaceutically acceptable salts can be found in the literature incorporated herein by reference [Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985].

[0102] Accordingly, when a therapeutically active agent, such as 4-chlorobutyl-CBN or its derivatives, but not limited thereto, or included in a pharmaceutical composition according to the present disclosure, has sufficiently acidic, sufficiently basic, or both sufficiently acidic and sufficiently basic functional groups, these groups or groups may therefore react with any number of inorganic or organic bases, inorganic and organic acids to form a pharmaceutically acceptable salt of 4-chlorobutyl-CBN. Examples of pharmaceutically acceptable salts are these salts prepared by the reaction of a pharmaceutically active compound with an inorganic acid, an organic acid, or an inorganic base, such as sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butin-1,4-dioate, hexine-1,6-dioate, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, It includes salts including methoxybenzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, tartrate, methane-sulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate.Where a pharmaceutically active compound has one or more basic functional groups, the desired pharmaceutically acceptable salt may be produced by any suitable method available in the art, for example, as an inorganic acid such as hydrochloric acid, hydrobromide, sulfuric acid, nitric acid, phosphoric acid, etc.; as an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvate, oxalic acid, glycolic acid, salicylic acid; as a pyranosidic acid such as glucuronic acid or galacturonic acid; as an alpha-hydroxy acid such as citric acid or tartaric acid; as an amino acid such as aspartic acid or glutamic acid; or as an aromatic acid such as benzoic acid or cinnamic acid. p - It can be prepared by treating a free base with a sulfonic acid, such as toluenesulfonic acid or ethanesulfonic acid. If the pharmaceutically active compound has one or more acidic functional groups, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary), an alkali metal hydroxide, or an alkaline earth metal hydroxide. Exemplary 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.

[0103] "Pharmaceuticalally acceptable" means that the salt, carrier, diluent, or excipient is compatible with other components of the composition and is not harmful to the recipient.

[0104] "Pharmaceutical excipients" refers to substances that aid in the administration of an active agent to a subject and / or absorption by the subject. Pharmaceutical excipients useful for the present disclosure include, but are not limited to, buffers, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, and colorings. A person skilled in the art will recognize other pharmaceutical excipients useful for the present disclosure.

[0105] In some cases, a protecting group may be included in the compound used in the method according to the present disclosure or in the compound according to the present disclosure. The use of such protecting groups may be used to prevent subsequent hydrolysis or other reactions that may occur in vivo and 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 acetyl, benzoyl, benzyl, β-methoxyethoxyethyl ether, dimethoxytrityl, methoxymethyl ether, methoxytrityl, p - Includes, but is not limited to, methoxybenzyl ether, methylthiomethyl ether, pivaloyl, tetrahydropyranyl, tetrahydrofuran, trityl, silyl ether, methyl ether, and ethoxyethyl ether. Protecting groups useful for protecting amines include carbobenzyloxy, p -Methoxybenzyl carbonyl, t -Butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, acetyl, benzoyl, benzyl, carbamate, p -Methoxybenzyl, 3,4-Dimethoxybenzyl, p - Includes methoxyphenyl, tosyl, trichloroethyl chloroformate, and sulfonamides. Protecting groups useful for protecting carbonyls include acetals, ketals, acilals, and dithianes. Protecting groups useful for protecting carboxylic acids include methyl esters, benzyl esters, tProtecting groups useful for protecting phosphate groups include butyl esters, 2,6-disubstituted phenol esters, silyl esters, ortho esters, and oxazolines. Protecting groups useful for protecting terminal alkynes 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.

[0106] As used herein, the term 'prodrug' refers to a derivative that is a precursor compound that releases a biologically active compound in vivo through certain chemical or physiological processes after administration (e.g., reaching a physiological pH or conversion of the prodrug into a biologically active compound via enzymatic action). The prodrug itself may lack or possess the desired biological activity. Therefore, the term 'prodrug' refers to a precursor of a pharmaceutically acceptable biologically active compound. In certain cases, the prodrug possesses improved physical and / or delivery properties compared to the parent compound from which it is derived. Prodrugs often offer advantages in solubility, histocompatibility, or delayed release in mammalian organisms (Reference [H. Bundgard, Design of Prodrugs (Elsevier, Amsterdam, 1988), pp. 7-9, 21-24]). Discussion of prodrugs is [T. Higuchi et al., "Pro-Drugs as Novel Delivery Systems," ACS Symposium Series, Vol.

[14] and provided in [EB Roche, ed., Bioreversible Carriers in Drug Design (American Pharmaceutical Association & Pergamon Press, 1987)]. Exemplary benefits of prodrugs may include, but are not limited to, their physical properties such as enhanced drug stability for long-term storage.

[0107] The term “prodrug” also means comprising any covalently bonded carrier that releases an active compound in vivo when the prodrug is administered to a subject. A prodrug of a therapeutically active compound as described herein may be prepared by modifying one or more functional groups present in a therapeutically active compound, including 4-chlorobutyl-CBN or 4-chlorobutyl-CBN derivatives, and other therapeutically active compounds used in the method according to the present disclosure or included in the pharmaceutical composition according to the present disclosure, in such a manner that the modification is cleaved in a conventional operation or in vivo to produce a parent therapeutically active compound. The prodrug comprises a compound in which a hydroxy, amino, or mercapto group is covalently bonded to any group that is cleaved to form a free hydroxy, free amino, or free mercapto group, respectively, when the prodrug of the active compound is administered to a subject. Examples of prodrugs include, but are not limited to, formate or benzoate derivatives of alcohol or acetamide, and formamide or benzamide derivatives of therapeutically active preparations having amine functional groups available for reaction. In some cases, the prodrug is a protected group modified derivative of a compound such as a protected group modified 4-chlorobutyl-CBN or a protected group modified derivative of 4-chlorobutyl-CBN.

[0108] Similarly, where the disclosed compound or a pharmaceutically acceptable form of the compound contains an alcohol functional group, the prodrug may be formed by replacing a 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, α-amino(C1-C4)alkanoyl, aryl acyl and α-aminoacyl, or α-aminoacyl-α-aminoacyl, wherein each α-aminoacyl group is a naturally occurring L-amino acid, P(O)(OH)2, P(O)(O(C1-C6)alkyl)2, or glycosyl (a hemiacetal form of a carbohydrate). It is independently selected from radicals resulting from the removal of hydroxyl groups.

[0109] The use of prodrug systems is discussed in the literature [T. This is described in et al., "Design and Pharmaceutical Applications of Prodrugs" in Drug Discovery Handbook (SC Gad, ed., Wiley-Interscience, Hoboken, NJ, 2005), ch. 17, pp. 733-796). Other alternatives for the composition and use of prodrugs are known in the art. Where the method or pharmaceutical composition according to the present disclosure uses or contains a prodrug of 4-chlorobutyl-CBN or other therapeutically active preparations, the prodrug and active metabolite of the compound can be identified using routine techniques known in the art. For example, in the literature [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).

[0110] As used herein, the terms 'therapeutic effective quantity', 'therapeutic effective dose', or 'therapeutic effective amount' refer to a dose of one or more compositions or pharmaceutical compositions described herein that produce the therapeutic effect to be administered. The precise dose will vary depending on the purpose of treatment and will be determined by a person skilled in the art using known techniques (see, for example, 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).

[0111] cannabinoids

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

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

[0114] In some cases, cannabinoids or their precursors may be formulated into tablets, derivatized (e.g., to form a prodrug or salt, or to form a target cannabinoid from the precursor), and / or pharmaceutical compositions.

[0115] Cannabinoids include, but are not limited to, phytocannabinoids. In some cases, cannabinoids include cannabinol, cannabidiol, Δ 9 -Tetrahydrocannabinol (Δ 9 -THC), non-natural cannabinoid HU-210(6a R , 10a R )-9-(hydroxymethyl)-6,6-dimethyl-3-(2-methyloctane-2-yl)-6 H , 6a H ,7 H , 10 H , 10a H -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 enantiomer of HU-308, cannabidivarin (CBDV), cannabichromene (CBC), cannabichromevarin (CBCV), cannabigerol (CBG), cannabigerovarin (CBGV), cannabielsoin (CBE), cannabicyclol (CBL), cannabivarin (CBV) and It includes, but is not limited to, cannabitriol (CBT). It includes other cannabinoids, including tetrahydrocannibivarin (THCV) and cannabigerol monomethyl ether (CBGM). Additional cannabinoids include cannabichromenic acid (CBCA), Δ 9 - Includes tetrahydrocannabinolic acid (THCA); and cannabidiolic acid (CBDA); and these additional cannabinoids are characterized by the presence of a carboxylic acid group in their structure.

[0116] Other cannabinoids include nabilone, limonabant, JWH-018 (naphthalene-1-yl-(1-pentylindole-3-yl)methanone), JWH-073 naphthalene-1-yl-(1-butylindole-3-yl)methanone, CP-55940 (2-[(1R,2R,5R)-5-hydroxy-2-(3-hydroxypropyl)cyclohexyl]-5-(2-methyloctane-2-yl)phenol), dimethylheptylpyran, HU-331 (3-hydroxy-2-[(1 R)-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 -[(1 S ,2 S ,4 R )-1,3,3-trimethylbicyclo[2.2.1]heptane-2-yl]-1 H -Pyrazole-3-Carboxamide), WIN 55,212-2((11 R )-2-methyl-11-[(morpholine-4-yl)methyl]-3-(naphthalene-1-carbonyl)-9-oxa-1-azatricyclo[6.3.1.0 4 ,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)-1 H It includes [-indole-3-yl]-(naphthalene-1-yl)methanone). Other cannabinoids include Δ 8 -Tetrahydrocannabinol (Δ 8 -THC), 11-hydroxy-Δ 9 -Tetrahydrocannabinol, Δ 11 - Includes 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 derivatizing it, for example, by a synthetic means. Non-natural cannabinoids include, but are not limited to, those described in U.S. Patent No. 9,394,267 by Attala et al., U.S. Patent No. 9,376,367 by Herkenroth et al., U.S. Patent No. 9,284,303 by Gijsen et al., U.S. Patent No. 9,173,867 by Travis et al., U.S. Patent No. 9,133,128 by Fulp et al., U.S. Patent No. 8,778,950 by Jones et al., U.S. Patent No. 7,700,634 by Adam-Worrall et al., U.S. Patent No. 7,504,522 by Davidson et al., U.S. Patent No. 7,294,645 by Barth et al., U.S. Patent No. 7,109,216 by Kruse et al., U.S. Patent No. 6,825,209 by Thomas et al., and U.S. Patent No. 6,284,788 by Mittendorf et al.

[0118] In one or more embodiments, the compound of the present disclosure is a cannabinoid. The cannabinoid according to the present disclosure may be at least partially selective for binding to either the CB2 cannabinoid receptor or the CB1 cannabinoid receptor. In one or more embodiments, the cannabinoid binds to both the CB1 and CB2 cannabinoid receptors. In some cases, the cannabinoid according to the present disclosure acts as a selective and partial agonist for the CB1 cannabinoid receptor. In some other cases, the cannabinoid according to the present disclosure acts as a selective and partial agonist for the CB2 cannabinoid receptor. In some cases, the cannabinoid binds to both the CB1 and CB2 receptors and acts as a partial agonist for both receptors, but Δ 9It has a higher affinity for CB2 receptors with efficacy similar to that of -THC. In some cases, one of the cannabinoids in a cannabinoid or mixture of cannabinoids is an antagonist or counter-agonist of CB1 and / or CB2 receptors. As an antagonist or counter-agonist, the cannabinoid may bind to CB1 and / or CB2 receptors but may induce a pharmacological response opposite to that of the agonist. In some cases, the cannabinoid in the pharmaceutical compositions and methods according to the present disclosure is partially selective for the CB2 cannabinoid receptor. In some cases, the cannabinoid or mixture of cannabinoids has at least a 3-fold lower K for the CB2 receptor compared to the CB1 receptor in an in vitro competitive assay having, for example, an overall higher binding affinity for CB2. i It can represent.

[0119] Pharmaceutical composition

[0120] The pharmaceutical compositions described herein are typically formulated for administration. Accordingly, pharmaceutical compositions comprising 4-chlorobutyl-CBN formulated for administration with one or more pharmaceutically acceptable carrier(s), diluent(s), or excipient(s) are described herein.

[0121] Pharmaceutical compositions can be manufactured by known procedures using well-known and readily available ingredients.

[0122] Pharmaceutical compositions containing 4-chlorobutyl-CBN may be formulated for administration to a subject in dose units containing a conventional non-toxic, pharmaceutically acceptable carrier, adjuvant, excipient, and / or vehicle by one of various standard routes, e.g., by eye, orally, topically, parenterally, by inhalation or spray, rectally, or vaginally.

[0123] As used herein, the term parenteral includes, in one or more embodiments, subcutaneous injection, intradermal, intra-articular, intravenous, intramuscular, intravascular, intrasternal, intra-spinal injection and infusion techniques. The pharmaceutical composition will typically be formulated in a form suitable for administration to a subject by a selected route, such as, for example, eye drops, ophthalmic depots, syrups, elixirs, tablets, troches, lozenges, hard or soft capsules, pills, suppositories, oily or aqueous suspensions, dispersible powders or granules, emulsions, injections, or solutions.

[0124] In one or more embodiments, the pharmaceutical composition is formulated for administration via a systemic route, e.g., intravenously, intramuscularly, intradermally, intraperitoneally, subcutaneously, or orally.

[0125] Pharmaceutical compositions for oral use may be prepared in solid or fluid unit dosage forms. Fluid unit dosage forms may be prepared according to procedures known in the art for the preparation of pharmaceutical compositions, and such pharmaceutical compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide pharmaceutically excellent and palatable formulations. Elixirs are prepared using a hydroalcoholic (e.g., ethanol) vehicle with a suitable sweetener, such as sugar or saccharin, together with an aromatic flavoring agent. Suspensions may be prepared with an aqueous vehicle with the help of a suspending agent, such as acacia, tragacanth, methylcellulose, etc. 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 contain 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.

[0126] Solid pharmaceutical compositions, including but not limited to tablets, gels, or chews, contain an active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for their manufacture. For example, these excipients may be inert diluents such as, for example, calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulators and disintegrants, for example, corn starch or alginate; binders, for example, starch, gelatin, or acacia; lubricants, for example, magnesium stearate, stearic acid, or talc; and other conventional ingredients, for example, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, methylcellulose, and functionally similar substances. Solid pharmaceutical compositions may not be coated, or they may be coated by known techniques. Among other reasons, coated solid pharmaceutical compositions, such as coated tablets, may delay degradation and absorption in the gastrointestinal tract, thereby providing sustained action over a long period. For example, time-delaying substances, such as glyceryl monostearate or glyceryl distearate, may be used.

[0127] The oral pharmaceutical composition may also be provided as a hard gelatin capsule in which the active ingredient is mixed with an inert solid diluent, e.g., calcium carbonate, calcium phosphate, or kaolin, or as a soft gelatin capsule in which the active ingredient is mixed with a water or oil medium, e.g., medium chain triglyceride oil (MCT oil), medium chain fatty acid (MCFA), liquid paraffin, coconut oil, palm kernel oil, olive oil, or in some cases, peanut oil. The soft gelatin capsule is prepared by mechanically encapsulating a slurry of the compound with an acceptable inert oil, e.g., vegetable oil, hard liquid petroleum jelly, MCT oil, MCFA oil, coconut oil, palm kernel oil, peanut oil, or other inert oil.

[0128] The aqueous suspension contains an active ingredient mixed with one or more excipients suitable for the preparation of the aqueous suspension. These excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and acacia gum; and includes a dispersant or wetting agent, such as a naturally occurring phosphatidide (e.g., lecithin), a condensation product of an alkylene oxide and a fatty acid (e.g., polyoxyethylene stearate), a condensation product of an ethylene oxide and a long-chain aliphatic alcohol (e.g., hepta-decaethyleneoxycetanol), a condensation product of a partial ester derived from a fatty acid and hexitol and ethylene oxide (e.g., polyoxyethylene sorbitol monooleate) or a condensation product of a partial ester derived from a fatty acid and hexitol anhydride and ethylene oxide (e.g., polyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl-p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, or one or more sweeteners, such as sucrose or saccharin.

[0129] Oily suspensions can be formulated by suspending the active ingredient in vegetable oil, e.g., peanut oil, olive oil, sesame oil, or coconut oil, or in mineral oil, e.g., liquid paraffin. Oily suspensions may contain thickeners, e.g., beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavoring agents such as those presented above may be added to provide a palatable oral formulation. These pharmaceutical compositions may be preserved by the addition of antioxidants such as ascorbic acid.

[0130] Dispersible powders and granules suitable for preparing an aqueous suspension by adding water provide an active ingredient mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or hydroxyl groups are exemplified by those already mentioned above. Additional excipients, such as sweeteners, flavoring agents, and coloring agents may also be present.

[0131] The pharmaceutical composition may also exist in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, e.g., olive oil or peanut oil, or a mineral oil, e.g., liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, e.g., acacia gum or tragacanth gum; naturally occurring phosphatides, e.g., soybeans, lecithin; and esters or partial esters derived from fatty acids and hexitol, anhydrides, e.g., sorbitan monooleate; and condensation products of these partial esters and ethylene oxide, e.g., polyoxyethylene sorbitan monooleate. The emulsion may also optionally contain sweeteners and flavoring agents.

[0132] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. Such suspensions may be formulated as known in the art using suitable dispersants or wetting agents and suspending agents, such as those mentioned above. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol, for example. Other acceptable vehicles and solvents that may be used include, for example, water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile fixing oils may be used as solvents or suspension media. Various laxative fatty oils known to be suitable for this purpose, including synthetic mono- or diglycerides, may be used. Furthermore, fatty acids such as oleic acid may find use in the manufacture of injectables. Adjuvants such as local anesthetics, preservatives, and buffers may also optionally be included in the injectable solution or suspension.

[0133] The pharmaceutical composition may include pharmaceutically acceptable excipients, 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 of 0.01 to 1% (w / v), for example, 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, for example, sucrose. In one or more embodiments, the sugar is present in an amount of 1 to 10% (w / v) of the buffer, for example, 2 to 8% (w / v), 4 to 6% (w / v), or 5% (w / v). In one or more embodiments, the buffer comprises a solvent, such as dimethyl sulfoxide (DMSO). In one or more embodiments, DMSO is present in an amount of 0.1 to 1% (w / v), such as 0.1 to 0.5% (w / v) or 0.2 to 0.3% (w / v) of the buffer.

[0134] Other pharmaceutical compositions and methods for preparing pharmaceutical compositions are known in the art, for example, in the literature ["Remington: The Science and Practice of Pharmacy" (formerly "Remington's Pharmaceutical Sciences")], [Gennaro, A., Lippincott, Williams & Wilkins, Philadelphia, Pa. (2000)].

[0135] The concentration of a compound (e.g., 4-chlorobutyl-CBN) in the pharmaceutical composition will vary depending on the condition to be treated and / or the method of administration.

[0136] method

[0137] A method for protecting neurons from neurodegenerative stimuli is described herein. Generally, the method comprises the step of contacting a neuron with a pharmaceutical composition comprising an effective amount of 4-chlorobutyl-CBN or its pharmaceutically acceptable salt, or 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or a 4-chlorobutyl-CBN derivative (e.g., a prodrug). The method may be an in vitro method. Alternatively, the method may be a method performed at least partially in vivo, such as by administering the pharmaceutical composition to a subject. Administration may be performed systemically (e.g., intravenously or subcutaneously) or by local injection. For example, local injection may include intravitreal injection. Administration may be performed by a non-invasive local administration method. For example, local administration to retinal neurons, such as retinal ganglia, may include the administration of ophthalmic pharmaceutical compositions such as hydrogels (e.g., see WO 2018 / 205022) or microemulsions (e.g., see U.S. Patents No. 8,968,775 and No. 9,149,453).

[0138] As described in US 8,968,775 and US 9,149,453, an ophthalmic pharmaceutical composition may comprise a pharmaceutical carrier suitable for topical administration to the eye 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, polysorbate and propylene glycol, polysorbate and glycerol, polysorbate and 1,2,3-triacetoxypropane, polyethoxylated castor oil and 1,2,3-triacetoxypropane, and polyethoxylated castor oil and propylene glycol; and (iii) an oil-in-water microemulsion comprising water, wherein water represents 50 to about 95 percent (w / w) of the pharmaceutical carrier, and the pair of oils and surfactants represents substantially all of the remainder of the pharmaceutical carrier, and the percentage of the pair of surfactants The ratio of (w / w) to oil percentage (w / w) is at least about 10:1.

[0139] The pharmaceutical composition may comprise a lipophilic active pharmaceutical ingredient [API] and the pharmaceutical carrier described above, wherein the pharmaceutical composition is formulated for topical administration to the eye.

[0140] In one or more embodiments, the ophthalmic pharmaceutical composition comprises a compound of Formula I.

[0141] In one or more embodiments, the compound or its pharmaceutical composition is administered for a period of less than 6 weeks. In one or more embodiments, the compound or its pharmaceutical composition is administered for a period of about 1 to 4 weeks. In one or more embodiments, such as for treating a neurodegenerative disease such as age-related macular degeneration, the compound or its pharmaceutical composition will be administered for an extended period, for example, for several years or for the remainder of the patient's life. The compound or its pharmaceutical composition may be administered every other month (once every two months), monthly, weekly, every other day, daily, twice a day, or three times a day.

[0142] The compound or its pharmaceutical composition may be administered to treat the eye of a subject requiring treatment to protect retinal neurons (e.g., optic 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. When the compound or its pharmaceutical composition is administered to protect neurons such as retinal neurons, the compound or its pharmaceutical composition may be administered in a dosage that provides a peak, median, or trough, preferably peak, neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous fluid, in 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 its pharmaceutical composition is administered to treat the eye of a subject requiring treatment to protect retinal tissue, such as the retina and / or retinal pigment epithelium (RPE). In one or more embodiments, the compound or its pharmaceutical composition is administered to treat the eye of a subject requiring treatment to protect photoreceptor cells (photoreceptors). In one or more embodiments, the compound or its pharmaceutical composition is administered to treat the eye of a subject requiring treatment to protect against or reduce the accumulation of autofluorescent extracellular debris. In one or more embodiments, the compound or its pharmaceutical composition is administered to treat the eyeball of a subject requiring treatment to protect against neuroinflammation (in some embodiments where the subject has AMD, in some embodiments where the subject has glaucoma, and in some embodiments where the subject does not have glaucoma).

[0143] In one or more embodiments, the neuroprotective effective concentration of a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye and / or in contact with ocular neuron 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 a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye, and / or in contact with ocular neuron cells is greater than about 0.15 μM and less than about 25 μM, or greater than 0.15 μM and less than 25 μM, or at least about 0.15 μM and less than about 25 μM, or at least 0.15 μM and less than 25 μM, or greater than about 0.15 μM and less than about 20 μM, or greater than 0.15 μM and less than 20 μM, or at least about 0.15 μM and less than about 20 μM, or at least about 0.15 μM and less than 20 μM.

[0144] In one or more embodiments, the neuroprotective effective concentration of a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye, and / or in contact with ocular neuron cells is greater than about 0.15 μM and less than about 15 μM, or greater than 0.15 μM and less than 15 μM, or at least about 0.15 μM and less than about 15 μM, or at least 0.15 μM and less than 15 μM, or greater than about 0.15 μM and less than about 12 μM, or greater than 0.15 μM and less than 12 μM, or at least about 0.15 μM and less than about 12 μM, or at least about 0.15 μM and less than about 12 μM, or at least about 0.15 μM and less than 12 μM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) is at least about 0.5 μM to less than about 25 μM, or at least 0.5 μM to less than 25 μM, or at least about 0.5 μM to less than about 20 μM, or 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 its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) is at least about 0.5 μM to less than about 15 μM, or at least 0.5 μM to less than 15 μM, or at least about 0.5 μM to less than about 12 μM, or at least 0.5 μM to less than 12 μM.

[0145] In one or more embodiments, the neuroprotective effective concentration of a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye and / or in contact with ocular neuron 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 a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye, and / or in contact with ocular neuron cells is greater than about 0.15 μM and less than about 25 μM, or greater than 0.15 μM and less than 25 μM, or at least about 0.15 μM and less than about 25 μM, or at least 0.15 μM and less than 25 μM, or greater than about 0.15 μM and less than about 20 μM, or greater than 0.15 μM and less than 20 μM, or at least about 0.15 μM and less than about 20 μM, or at least about 0.15 μM and less than 20 μM.

[0146] In one or more embodiments, the neuroprotective effective concentration of a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye, and / or in contact with ocular neuron cells is greater than about 0.15 μM and less than about 15 μM, or greater than 0.15 μM and less than 15 μM, or at least about 0.15 μM and less than about 15 μM, or at least 0.15 μM and less than 15 μM, or greater than about 0.15 μM and less than about 12 μM, or greater than 0.15 μM and less than 12 μM, or at least about 0.15 μM and less than about 12 μM, or at least about 0.15 μM and less than about 12 μM, or at least about 0.15 μM and less than 12 μM. In one or more embodiments, the neuroprotective effective concentration of the compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) is at least about 0.5 μM to less than about 25 μM, or at least 0.5 μM to less than 25 μM, or at least about 0.5 μM to less than about 20 μM, or 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 its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) is at least about 0.5 μM to less than about 15 μM, or at least 0.5 μM to less than 15 μM, or at least about 0.5 μM to less than about 12 μM, or at least 0.5 μM to less than 12 μM.

[0147] In one or more embodiments, the neuroprotective effective concentration of a compound (e.g., 4-chlorobutyl-CBN or its pharmaceutically acceptable salt or its derivative (e.g., a prodrug)) in the vitreous humor, in the ocular tissue of the eye, and / or in contact with ocular neuron cells is greater than about 0.15 μM and less than about 10 μM, or greater than 0.15 μM and less than 7.5 μM, or at least about 0.15 μM and less than about 10 μM, or at least 0.15 μM and less than 7.5 μM, or greater than about 0.15 μM and about 5 μM, or greater than 0.15 μM and 5 μM, or at least about 0.15 μM and about 5 μM, or at least about 0.15 μM and about 5 μM.

[0148] For example, the compound or its pharmaceutical composition may be administered orally, intrathecally, intravenously, topically, or by injection, or may be administered directly to a site of a target neuron or a target neuron population. In one or more embodiments, a neuroprotective effective concentration is achieved by systemic doses of about 1 mg / kg to about 100 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, or about 1 mg / kg to about 13 mg / kg. The dose may be repeated, for example, every other month, every month, every week, every other day, daily, or twice daily.

[0149] The compound or its pharmaceutical composition may be administered intrathecally, intravenously, or by injection, or may be administered directly into the eye, such as by topical ocular drops, intravitreal injection, or by a pump. In one or more embodiments, for ocular administration in ocular indications (e.g., for treating age-related macular degeneration), the systemic dose may be 1 mg / kg to 20 mg / kg. The dose may be repeated, for example, every other month, monthly, weekly, every other day, daily, or twice daily. In one or more embodiments, for systemic administration in ocular indications (e.g., for treating age-related macular degeneration), the systemic dose may be 1 mg / kg to 15 mg / kg, 1 mg / kg to 13 mg / kg, or 1 mg / kg to 10 mg / kg. The dose may be repeated, for example, every other month, monthly, weekly, every other day, daily, or twice daily. In one or more embodiments, for systemic administration in peripheral indications (e.g., for treating peripheral neuropathy and / or peripheral nerve injury or damage), the dose may be 1 mg / kg to 20 mg / kg, 1 mg / kg to 15 mg / kg, 1 mg / kg to 13 mg / kg, or 1 mg / kg to 10 mg / kg. The dose may be repeated, for example, every other month, every month, every week, every other day, daily, or twice a day. In one or more embodiments, for systemic administration in central indications (e.g., for treating central nerve injury or damage), the dose may be 1 mg / kg to 20 mg / kg, 1 mg / kg to 15 mg / kg, 1 mg / kg to 13 mg / kg, or 1 mg / kg to 10 mg / kg. The dose may be repeated, for example, every week, daily, or twice a day.

[0150] In one or more embodiments, for ocular administration in ocular indications (e.g., for treating age-related macular degeneration), the ocular dose may be 0.5 mg to 20 mg, 0.5 mg to 15 mg, 0.5 mg to 10 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 10 mg, 0.5 mg to 5 mg, or 1 mg to 5 mg applied to the eye in the form of eye drops or eye gel. The dose may be repeated, for example, every other month, every month, every week, every other day, daily, or twice a day. In one or more embodiments, for ocular administration in ocular indications (e.g., for treating age-related macular degeneration), the ocular dose may be 0.05 mg to 2 mg, 0.05 mg to 1.5 mg, 0.05 mg to 1 mg, 0.1 mg to 2 mg, 0.1 mg to 1.5 mg, 0.1 mg to 1 mg, 0.05 mg to 0.5 mg, or 0.1 mg to 0.5 mg applied to the eye, such as in the form of an intravitreal injection or a pump. The dose may be repeated, for example, every other month, every month, every week, every other day, daily, or twice a day.

[0151] In one or more embodiments, the compound or its pharmaceutical composition is administered within about 0 to 48 hours after the occurrence of injury affecting retinal neurons, photoreceptors, retinal pigment epithelium, neuroinflammation and / or the accumulation of autofluorescent extracellular deposits. In one or more embodiments, the compound or its pharmaceutical composition is administered within about 2 to 24 hours after the occurrence of injury affecting retinal neurons, photoreceptors and / or retinal pigment epithelium. In one or more embodiments, the compound or its pharmaceutical composition is administered within about 3 to 12 hours after the occurrence of injury affecting retinal neurons, photoreceptors and / or retinal pigment epithelium. In one or more embodiments, the compound or its pharmaceutical composition is administered within about 3 to 5 hours after the occurrence of injury affecting retinal neurons, photoreceptors and / or retinal pigment epithelium.

[0152] In one or more embodiments, the compound or its pharmaceutical composition is administered to a subject having AMD.

[0153] The method may include, or further include, the step of administering an additional activator (e.g., a second drug) simultaneously or sequentially in combination with the compound or its pharmaceutical composition. In some cases, the additional activator is a therapeutic agent for the treatment of age-related macular degeneration. In one or more embodiments, the additional activator is an anesthetic. In one or more embodiments, the additional activator is a VEGF inhibitor. In one or more embodiments, the additional activator is an anti-VEGF protein or compound. For example, the method may include the step of anesthetizing a subject before administering the compound or pharmaceutical composition of one or more embodiments. In other examples, the subject is anesthetized prior to contact (of the administration step) in the method of one or more embodiments.

[0154] Examples

[0155] Materials and Methods

[0156] Animals and experimental groups

[0157] Sprague Dawley (SD) albino rats were born and raised under dark periodic light conditions (12 hours light, 12 hours dark) with an ambient light level of approximately 5 lux. The animals were free to consume food and water. The experiment was conducted on adult animals aged 2 to 4 months. The therapeutic agent was administered via intravitreal injection to both eyes the day before photoinjury, and possible retinal protection was evaluated 7 days after photoinjury using fERG recordings.

[0158] Preparation of pharmaceutical compositions and intravitreal injection

[0159] 4-chlorobutyl-CBN was prepared by combining biosynthesis and chemical synthesis. Briefly, 5-chlorovaleric acid was 'supplied' to a Saccharomyces cerevisiae strain engineered to convert an acid analog into 4-chlorobutyl-olivetolic acid.

[0160]

[0161] 4-chlorobutyl-olivetolic acid analogs were extracted with ethyl acetate and recovered from the culture medium. From 4-chlorobutyl-olivetolic acid, 4-chlorobutyl-CBN end products were produced using two different pathways. In the case of pathway 'A', 4-chlorobutyl-olivetolic acid was decarboxylated to 4-chlorobutyl-olivetolic acid, and

[0162]

[0163] Subsequently, 4-chlorobutyl-CBC was produced by combining with citral (chemical prenylation), and

[0164]

[0165] This was oxidized to 4-chlorobutyl-CBN. In a separate pathway ('B'), 4-chlorobutylolivetolic acid was engineered to overproduce the geranyl-diphosphate precursor (GPP) and express a heterologous gene encoding prenyltransferase and synthase.S. It was 'supplied' to the Cerevisiae strain. The biotransformed strain was converted from a 4-chlorobutyl-olivetolic acid analog to 4-chlorobutyl-THCA, and

[0166]

[0167] This was recovered from yeast cells by ethyl acetate extraction. Subsequently, 4-chlorobutyl-THCA was decarboxylated and oxidized to produce 4-chlorobutyl-CBN. 4-chlorobutyl-CBN can also be prepared through chemical synthesis. Additionally, a method similar to that described in US-2023-0063396-A1 may be used or available for the preparation of 4-chlorobutyl-CBN.

[0168] 15 mM or 30 mM stock solutions of 4-Cl-butyl CBN in 100% DMSO were diluted in sterile dilution buffer (0.03% polysorbate (Tween) 20 and 5% sucrose in PBS) to achieve final formulation concentrations of ~100 μM, 30 μM, and 10 μM at final DMSO concentrations of 0.2 to 0.4% for intravitreal injection administration. Formulations were prepared by serial dilution to ensure similar levels of DMSO were included at different dose levels. Buffers were prepared by dissolving 0.03% polysorbate (Tween) 20 and 5% sucrose in PBS buffer. For the buffer solutions used for the vehicle(s) and control(s) (or injection formulations), the buffer solutions were diluted with DMSO to appropriate levels (to match those used in the stock solution(s) of the corresponding concentration / dilution, except for the absence of cannabinoids / APIs). Regarding DMSO in the buffer, the injectable formulation(s) contained DMSO derived from a 4-chlorobutyl-CBN stock solution; for example, the stock was diluted to 100 μM when injected into the eye, and the 4-chlorobutyl-CBN concentration was 10 μM. Therefore, DMSO was added to the buffer solutions for the vehicle and control experiments to match the DMSO concentration of the corresponding injectable formulation(s). The buffer solutions were used for the vehicle(s) and control(s) described below. After dissolving the excipients, the buffer solution was filtered through a 0.2 μM filter to prepare a sterile buffer for formulation preparation. The DMSO stock solution was diluted in the buffer solution to achieve final formulation concentrations of ~100 μM, 30 μM, and 10 μM. Subsequently, vortex mixing was performed at a high setting for at least 30 seconds. The final concentration in the vitreous fluid was 1 / 10 of the injection concentration. For example, 1 / 10 of an initial concentration of 100 μM was a final concentration of 10 μM (see figure), and likewise, 1 / 10 of an initial concentration of 30 μM was a final concentration of 3 μM (see figure), and also, 1 / 10 of an initial concentration of 3 μM was 0.The final concentration was 3 μM (see figure), and others. The final expected concentrations achieved in the eye for the buffer components were 0.003% polysorbate (tween) 20, 0.5% sucrose, and ~0.02% to 0.04% DMSO.

[0169] For the surgical procedure, rats were anesthetized by intraperitoneal injection of ketamine / xylazine (10 mg / 100 g to 1.2 mg / 100 g). 2.0 μL of the compound was injected into both eyes using a Hamilton syringe under completely aseptic conditions (2). Control group: Healthy animals without photoinjury (LD) and without treatment. Vehicle group: Photoinjury and treated with dilution buffer. The vehicle group did not induce any change in the fERG response compared to the uninjected photoinjury eye. Therefore, the photoinjury group was not included in the analysis.

[0170] light damage

[0171] The animals were placed in individual plexiglass cages, with food placed on the floor and water in plastic bottles. After undergoing a dark acclimatization process overnight, they were exposed to 1,000 lux for 24 hours starting at 9 a.m. (LD24h). Subsequently, they were returned to dark periodic light conditions for 7 days to recover from acute stress caused by light exposure.

[0172] electroretinogram recording

[0173] fERG was recorded under dark-adapted conditions in response to a single flash of increasing intensity white light (dark adaptation) delivered by a standard Ganzfeld stimulator (Biomedica Mangoni, Pisa, Italy). Before performing fERG, rats were anesthetized by intraperitoneal injection of ketamine / xylazine (10 mg / 100 g to 1.2 mg / 100 g), mounted in a stereofixation device, and their body temperature was maintained at 37.5 °C. The cornea was anesthetized with one drop of procaine (Novocaine), and the pupils were dilated with Visumidriatic 1.0% Tropicamide. Recording was performed simultaneously on both eyes by placing a gold electrode loop (2.0 mm diameter) on the cornea. The reference electrode was inserted subcutaneously near the eye, and the ground electrode was inserted into the anterior scalp between the eyes. The response was recorded at increasing light intensities (ranging from 0.001 cd*s / m2 to 100 cd*s / m2). At the end of the recording session, the trace was band-pass filtered between 0.3 and 500 Hz. The a-wave amplitude (μV) and b-wave amplitude (μV) for individual luminances were measured. An oscillating potential (OP) analysis was also performed by calculating the sum of the amplitudes of OP1, OP2, OP3, and OP4.

[0174] Statistical analysis

[0175] For statistical analysis, one-way ANOVA and subsequent Dunnett post-hoc comparisons were performed between the CBN, 4-chlorobutyl-CBN, and vehicle groups, and for the analysis excluding the CBN group, Student between the 4-chlorobutyl-CBN and vehicle groups t - A test was performed. Statistical analysis was performed using SigmaPlot 12.0 software (Systat Software, San Jose, California, USA). Data are plotted as mean ± standard error of the mean. (4-Cl-CBN: n=14; CBN 30: n=17; CBN 100: n=13; Vehicle: n=17) (*p<0.05 relative to Vehicle).

[0176] Example 1: In vivo intravitreal injection, 0.3 μM of 4-chlorobutyl-CBN

[0177] In a separate experiment, in vivo studies were conducted in an AMD rat model by intravitreal injection of 3 μM of 4-chlorobutyl-CBN in the vehicle (final intraocular concentration 0.3 μM) and 3 μM of CBN in the vehicle (final intraocular concentration 0.3 μM). The vehicle alone and the control group were tested and compared separately. Intravitreal injections were 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.

[0178] As can be seen in Figures 2a and 2b, CBN at a final concentration of 0.3 μM showed superior performance compared to 4-chlorobutyl-CBN at a final concentration of 0.3 μM. However, Student between the CBN and 4-Cl-CBN groups t - The test result is p=0.46, so there is no statistically significant difference between the two groups. In this study, 4-chlorobutyl-CBN at a final concentration of 0.3 μM was tested on a smaller number of animals. Therefore, these results may not be statistically significant.

[0179] Example 2: In vivo intravitreal injection, 3 μM of 4-chlorobutyl-CBN

[0180] In a separate experiment, in vivo studies were conducted in AMD rat models via intravitreal injection of 4-chlorobutyl-CBN 30 μM (3 μM final), CBN 30 μM (3 μM final), and CBN 100 μM (10 μM final). Vehicle alone and a control group were tested and compared separately. Intravitreal injections were performed at the back of the eye, as shown in Fig. 1 (arrow). The results of this study are illustrated in Figs. 3a to 3f (a-wave amplitude), Figs. 4a to 4e (b-wave amplitude), and Fig. 5a (OP amplitude).

[0181] The α-wave amplitude represents the response of photoreceptors to light stimulation. As shown in Figures 3a to 3f, 4-chlorobutyl-CBN at 3 μM (final concentration) showed superior performance with respect to an increase in α-wave amplitude (μV) at luminance (light intensity) of 0.01, 0.1, 1, 10, and 100 cd*s / m^2 compared to CBN at 3 μM (final concentration) and CBN at 10 μM (final concentration). From these results, a statistically significant increase in α-wave amplitude was observed at intensities of 0.01, 0.1, and 100 cd*s / m^2.

[0182] b-wave amplitude represented the overall response of the inner retina to increasing light stimulation. As shown in Figures 4a to 4e, 4-chlorobutyl-CBN at 3 μM (final concentration) showed superior performance with respect to an increase in b-wave amplitude (μV) at luminance (light intensity) of 0.001, 0.01, 0.1, 1, 10, and 100 cd*s / m^2 compared to CBN at 3 μM (final concentration) and CBN at 10 μM (final concentration). From these results, a statistically significant increase in b-wave amplitude was observed at intensities of 0.1, 1, 10, and 100 cd*s / m^2.

[0183] The oscillatory potential (OP) amplitude represented the response of the inner retinal circuit to light stimulation (primarily due to rod bipolar and A11 / A17 amacrine cells). As shown in Fig. 5, 4-chlorobutyl-CBN at 3 μM (final concentration) showed superior performance for OP amplitude (μV) at luminance (light intensity) of 0.001, 0.01, 0.1, 1, and 10 cd*s / m^2 compared to CBN at 3 μM (final concentration) and CBN at 10 μM (final concentration).

[0184] Example 3: Imaging

[0185] At the end of electroretinogram recording, the eye was excised for morphological analysis, fixed in 4% paraformaldehyde for 6 hours, and washed with 0.1 M phosphate-buffered saline (PBS, pH 7.4). The cornea and lens were removed, and the remaining eye cup was immersed in 30% sucrose for cryoprotection. They were then embedded in TissueTek OCT (optimal cutting temperature, Qiagen, Valencia, California, USA), frozen, and stored at -20 °C. Subsequently, 20 μm thick frozen sections were obtained using a Leica CM1850 cryostat (Nussloch, GmbH, Germany) and collected on gelatin and poly-l-lysine coated slides. Sections transversing the optic nerve were selected for subsequent immunofluorescence analysis.

[0186] Immunofluorescence was used to identify microglia, astrocytes, and Müller cells. Specifically, non-specific binding sites were blocked using 5% bovine serum albumin (BSA) at room temperature (RT) for 1 hour. Subsequently, sections were incubated overnight at 4 °C with primary antibodies for the detection of microglia (IBA-1 antibody) (Wako Chemicals, #019-19741), astrocytes, and Müller cells (GFAP antibody) (Cell Signaling, #3670). Then, sections were incubated with a secondary antibody (anti-mouse or anti-rabbit IgG conjugated with a red or green fluorescent dye) diluted 1:300 (Alexa Fluor 594 or 488; Molecular Probes, Invitrogen, Carlsbad, California, USA) and incubated at 37 °C for 2 hours. The nuclei were labeled using a bisbenzimide nuclear dye (Hoechst) and the retinal layers were identified. Images were then obtained using a fluorescence microscope. The results are shown in Fig. 6a. In Fig. 6b, a negative image is provided to show contrast.

[0187] The photodamage model of AMD is characterized by thinning of the outer layer where photoreceptors are located. Generally, this is a characteristic of photoreceptor degeneration associated with AMD. Along with photoreceptor death, glial phenomena occur in the retina, resulting in (i) infiltration of microglia (present in the inner retina under physiological conditions) into the outer layer and (ii) responsiveness of astrocytes and Müller cells characterized by the upregulation of GFAP markers.

[0188] Based on this, analysis of retinal cryosections investigated whether the aforementioned adverse phenomena were prevented by the administration of 3 μM of 4-Cl-butyl CBN. Overall, morphological analysis indicated that 4-Cl-butyl CBN can prevent photoreceptor apoptosis and gliosis in photodamaged retinas.

[0189] Specifically, the treatment provided the following protective results.

[0190] (i) Increased outer core layer thickness compared to the vehicle,

[0191] (ii) Inhibition of IBA-1 (+) cell invasion in the outer nuclear layer, and

[0192] (iii) Attenuation of GFAP expression across the entire retinal layer compared to the vehicle.

[0193] Example 4. Comparison between 4-chlorobutyl-CBN and control animals

[0194] To better evaluate the degree of functional protection provided by 4-chlorobutyl-CBN in a photodamage model, Student [student] compared 4-chlorobutyl-CBN-treated eyes with controls (eyes of healthy animals) t - A test analysis was performed. As shown in the table, 12 of the 18 investigated fERG conditions were not significantly different between the control group and the treated eyes.

[0195] Table 1: Between 4-chlorobutyl-CBN and control animals t - Test comparison

[0196]

[0197] The results indicate a tendency for 4-chlorobutyl-CBN (e.g., a final concentration of 3 μM as described above) to significantly prevent photo-induced retinal functional impairment compared to the vehicle, due to alpha-wave, alpha-wave, and oscillatory potential (OP) amplitudes. In general, 4-chlorobutyl-CBN preserved fERG values ​​similar to those of healthy, uninjured animals. Functional protection by 4-chlorobutyl-CBN was evident for alpha-wave and OP, suggesting greater protection of the inner retina compared to the outer retina. Additionally, slight improvement in retinal function occurred in response to CBN (when used at 3 μM) at 100 cd*s / m^2. That is, in these tests, there was little to no difference between the control group (healthy eyes, not exposed to light) and the eyes treated with 4-chlorobutyl-CBN upon light exposure.

[0198] Example 5: In vivo concentration

[0199] 4-Cl-butyl-CBN was tested at concentrations of 0.3 μM, 1 μM, 3 μM, and 10 μM (intraocular). The alpha-wave amplitude, representing the photoreceptor response, was improved by 4-Cl-butyl-CBN at concentrations of 3 μM and 10 μM compared to the vehicle, indicating that the treatment exerted neuroprotection on photoreceptors against photoinjury. Control group: Healthy animals without photoinjury and receiving no treatment. Vehicle group: Animals with photoinjury and treated with dilution buffer. The results of these tests are shown in Fig. 7a.

[0200] 4-Cl-butyl-CBN was tested at 0.3 μM, 1 μM, 3 μM, and 10 μM (intraocular). The b-wave amplitude, representing the overall response of the inner retina after light stimulation, was improved by 4-Cl-butyl-CBN at concentrations of 3 μM and 10 μM compared to the vehicle, indicating that the treatment prevented retinal functional impairment. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle group: Animals with photodamage treated with dilution buffer. The results of these tests are shown in Fig. 7b.

[0201] 4-Cl-butyl-CBN was tested at concentrations of 0.3 μM, 1 μM, 3 μM, and 10 μM (intraocular). The oscillatory potential (OP) amplitude, which represents the function of the inner retinal circuit, was improved by 4-Cl-butyl-CBN at concentrations of 3 μM and 10 μM compared to the vehicle, indicating that the treatment exerted retinal neuroprotection and prevented inner retinal dysfunction. Control group: Healthy animals without photodamage and receiving no treatment. Vehicle: Animals with photodamage and treated with dilution buffer. The results of these tests are shown in Fig. 7c.

[0202] Electrophysiological results obtained from photo-damaged animals treated with 4-Cl butyl CBN and CBN via intravitreal injection demonstrated that 4-Cl butyl CBN at a concentration of 3 μM exhibited superior functional protection compared to vehicle-injected eyes, which is greater than that of CBN at the same concentration, as can be seen in Figures 3a and 4a.

[0203] Example 6: Histology and Molecular Analysis

[0204] Representative retinal sections from all experimental groups were stained with the nuclear dye Hoechst (blue) as shown in Fig. 8a. The control group refers to healthy rats without photodamage. The vehicle group refers to animals with photodamage (disease) and is treated with injection buffer.

[0205] The red line highlights the outer nuclear layer thickness (ONL). The ratio of ONL to retinal thickness was determined as shown in Fig. 8b, where Fig. 8c illustrates specific locations on the superior and inferior retinas where the thickness ratio was determined relative to Fig. 8b. These data demonstrate that 4-Cl-butyl-CBN improved the outer nuclear layer thickness in the AMD model.

[0206] Autofluorescence (AF) extracellular deposits were evaluated on retinal cryosections using an excitation wavelength of 594 nm as shown in Fig. 9a (AF deposits are shown in red). The number of AF deposits was counted across the retinal sections as shown in Fig. 9b. These data show that 4-Cl-butyl-CBN reduced extracellular deposits in the AMD model.

[0207] Retinal pigment epithelial (RPE) integrity was evaluated in retinal frozen sections immunostained with anti-RPE65 as shown in Fig. 10a (immunostaining is indicated in red). Frozen sections crossing the optic nerve were selected for analysis, and RPE integrity was measured in the dorsal retina. White segments depict intact RPE. RPE65 (+) length was measured as shown in Fig. 10b. These data demonstrate that 4-Cl-butyl-CBN improved RPE length in the AMD model.

[0208] Photodamage models exhibit loss of RPE integrity, including loss of RPE cell-to-cell connectivity integrity (references [Mirales de Imperial-Ollero, et al. 2021], [Cachafeiro, et al. 2013] and [Geiger, et al. 2015]).

[0209] Microglia were labeled with anti-IBA-1 (green) in retinal cryosections from all experimental groups, as shown in Fig. 11a. IBA-1 (+) cells were counted in the outer retina (subretinal, outer nucleus, outer reticular layer) from the upper to the lower edge of the retina, as shown in Fig. 11b. The locations on the upper and lower retinas where data were collected are the same as those shown in Fig. 8c. The number of IBA-1 (+) cells in the upper retina, lower retina, and the entire retina (upper and lower) is shown in Figs. 11c through 11e, respectively. These data demonstrate that 4-Cl-butyl-CBN reduced inflammation in the retina in the AMD model.

[0210] Example 7: Receptor Interaction Study

[0211] The interactions between 4-Cl-butyl-CBN and different receptors were evaluated using various analytical methods. The analyses were performed using Eurofins' proprietary standard bioassay protocols.

[0212] The interactions between CB1 and CB2 receptors and 4-Cl-butyl-CBN were evaluated using the HitHunter(R) cAMP assay with cAMP as an analytical marker. Briefly, cAMP Hunter cell lines expressing either the CB1 or CB2 receptor were evaluated using the DiscoverX HitHunter cAMP XS+ assay. To evaluate agonist behavior (Gs agonist form), cells were cultured with the sample to induce a reaction; then, the medium was aspirated and replaced with HBSS / Hepes buffer containing cAMP XS+ Ab reagent; subsequently, 5 μL of the 4-fold sample was added to the cells, and 37 at a 1% vehicle concentration oIncubated at C or room temperature for 30 or 60 minutes. To evaluate agonist behavior (Gi agonist form), cells were cultured in the presence of EC80 forskolin to induce a reaction, then the medium was aspirated and replaced with HBSS / Hepes buffer containing cAMP XS+ Ab reagent, then 5 μL of a mixture of 4-fold sample and 4-fold EC80 forskolin was added to the cells, and 37 at a vehicle concentration of 1% o Incubated at C or room temperature for 30 or 60 minutes. To evaluate antagonist behavior, cells were pre-cultured with the sample, then stimulated with the agonist at an EC80 concentration, the medium was aspirated and replaced with HBSS / Hepes buffer containing cAMP CS+ Ab reagent, then 5 μL of the sample compound at a 4-fold concentration was added to the cells, and 37 o Incubate at C or room temperature for 30 minutes, then add 5 μL of EC80 agent and 37 o Incubated at C or room temperature for 30 or 60 minutes with a 1% vehicle concentration. For GI-binding GPCRs, EC80 forskolin was included in the antagonist assay. For signal detection in both assays, 20 μL of cAMP XS+ ED / CL lysis cocktail was added and incubated for one hour, followed by the addition of 20 μL of cAMP CS+ EA reagent and incubation at room temperature for three hours. Microplates were read for chemiluminescence detection.

[0213] The interaction between CB1 and CB2 receptors and 4-Cl-butyl-CBN was evaluated using the PathHunter(R) β-arrestin assay from Eurofins DiscoverX, using arrestin as an analysis marker.

[0214] The interactions between PPARγ, PPARα, and PPARδ and 4-Cl-butyl-CBN were evaluated using the PathHunter(R) Nuclear Hormone Receptor Protein Interaction (Pro) assay. Briefly, for agent determination, PathHunter NHR cell lines were cultured with the sample, then 5 μL of the 5-fold concentration sample was added to the cells, and 37 at a final vehicle assay concentration of 1% o Cells were incubated at C or room temperature for 3 to 16 hours. For antagonist determination, cells were pre-cultured with the antagonist, then stimulated with the agonist at an EC80 concentration, followed by the addition of 5 μL of the 5-fold concentration sample to the cells and 37 for 60 minutes at a final vehicle assay concentration of 1%. o After incubating at C or room temperature, add 5 μL of the EC80 agent to the analysis buffer and incubate for 3 to 16 hours. o Incubated at C or room temperature. 12.5 or 15 μL of PathHunter detection reagent cocktail was added and incubated at room temperature for 1 hour, after which the microplate was read for chemiluminescence detection to generate an analysis signal.

[0215] A similar analysis was performed on the GPR55 receptor β-arrestin pathway.

[0216] Data on receptor interactions are provided in Table 2 below.

[0217] Table 2: 4-Cl-butyl-CBN receptor interaction data

[0218]

[0219] These data show that 4-Cl-butyl-CBN is a selective (biased) CB1 and CB2 agonist and antagonist and a 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.

[0220] Example 8: 4-Cl-butyl-CBN levels in the retina after topical and IVT delivery

[0221] Topical ophthalmic formulations of 4-Cl-butyl-CBN were administered topically to rabbits in two formulations and compared with IVT levels in rats as shown in Figure 13.

[0222] IVT delivery in rats was performed in the disease model described in Example 1. Twenty-four hours after IVT injection, the animals were euthanized, and retinal / RPE / choroidal tissues were collected from each animal. Compound concentrations were measured using a validated LC-MS / MS bioassay. The microemulsion formulation for topical delivery in rabbits was based on the method previously described in U.S. Patents No. 8,968,775 and No. 9,149,453. In the topical delivery study performed in rabbits, animals received a topical dose of 40 μL (1% formulation) twice daily in both eyes for eight days. On day eight, the animals were euthanized, and retinal / RPE / choroidal tissues were collected from each animal. Compound concentrations were measured using a validated LC-MS / MS bioassay.

[0223] Example 9: Local delivery of 4-Cl-butyl-CBN to the retina in a Sprag-Dowry rat model

[0224] Topical administration of the compound was performed in Sprag Dauri albino rats via infusion of eye drops (10 μl) twice daily (at approximately 12-hour intervals) for 7 days prior to photoinjury, and continued for an additional 7 days after injury. At the end of the study, retinal function was evaluated via fERG recording, and RPE / retinal protection was investigated through histological evaluation of RPE integrity, accumulation of autofluorescent deposits, and retinal thickness.

[0225] Example 10: In vivo comparative analysis of AMD treatment with 4-Cl butyl CBN versus CBN

[0226] Ocular tissue is taken from 4-Cl butyl CBN or CBN-treated rats for further morphological and molecular analysis of AMD pathology as described below.

[0227] 1. Protection of the retinal pigment epithelium (RPE)

[0228] Retinal pigment epithelial cell death is a component of AMD pathology (reference [Kim et al., 2021]) and is also reproduced in photodamage models (reference [Jaadane et al., 2017], [Tisi et al., 2020a]). The efficacy of 4-Cl butyl CBN or CBN compounds in preventing retinal pigment epithelial degeneration is evaluated in retinal frozen sections immunolabeled with a selective RPE cell marker (RPE65), which allows for the assessment of the maintenance of retinal pigment epithelial layer integrity by treatment.

[0229] 2. Photoreceptor (PR) Neuroprotection

[0230] Photoreceptor cell death is another phenomenon of AMD (Mitchell et al., 2018) and is induced by photodamage in albino rats. Photoreceptor neuroprotection is investigated by quantifying the outer nucleus layer thickness and measuring the length of degenerated regions on retinal cryosections stained with bisbenzimide nuclear dye (Tisi et al., 2019). A larger outer nucleus layer thickness and a reduced length of degenerated regions are indicators of photoreceptor protection.

[0231] 3. Accumulation of extracellular deposits

[0232] Another AMD phenomenon is the accumulation of extracellular material (i.e., drusen and lipofuscin) visible through fundus imaging (references [Ach et al., 2014], [Davis et al., 2005]). Additionally, in photoinjury models, autofluorescent (AF) extracellular debris accumulates along with degeneration of the retinal pigment epithelium and photoreceptors (references [Tisi et al., 2020b]). Therefore, autofluorescent extracellular debris is quantified in retinal cryosections via fluorescence microscopy, and a reduction in autofluorescent extracellular debris deposits in the treated eye is an indicator of retinal protection.

[0233] 4. Neuroinflammation

[0234] Neuroinflammation is a major cause of AMD. Retinal neuroinflammation is associated with the reactivity of microglia and macroglia (astrocytes and Müller cells) (reference [Gupta et al., 2003], [Dhodapkar et al., 2022]). We investigate neuroinflammation in terms of IBA-1 (+) cell and GFAP expression across retinal layers using glial markers (IBA-1 for microglia and GFAP for astrocytes and Müller cells). A decrease in microglia number and GFAP expression in photodamaged retinas is an indicator of reduced neuroinflammation.

[0235] The aforementioned data further demonstrate neuroprotection of photoreceptors, improvement of photoreceptor function, reduction of extracellular AF deposits, and preservation of RPE integrity in a photo-damaged in vivo rat model. Additionally, 4-Cl-butyl-CBN represents a systemic compound across the blood-brain barrier and can be delivered via topical ophthalmic formulations or IVT formulations.

[0236] References

[0237] Ramin, et al. “Age-Related Macular Degeneration: A Scientometric Analysis,” Med Hypothesis Discov. Innov. Ophthalmol. 2015 Summer; 4(2): 39-49.

[0238] Apte, “Age-Related Macular Degeneration,” N. Eng. J. Med. 2021 Aug. 5; 385(6): 539-547.

[0239] 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.

[0240] 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.

[0241] 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.

[0242] 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.

[0243] Jaadane I, Villalpando Rodriguez GE, Boulenguez P, Chahory S, Carr 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.

[0244] 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.

[0245] 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.

[0246] 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.

[0247] 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.

[0248] 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.

[0249] 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.

[0250] 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.

[0251] 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.

[0252] 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.

[0253] 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.

[0254] * * *

[0255] The invention described herein by way of example may be suitably practiced without any elements or elements, or any limitations or restrictions, that are not specifically disclosed herein. Accordingly, terms such as, for example, “comprising,” “comprising,” and “containing” should be read broadly and without limitation. Additionally, the terms and expressions used herein are descriptive rather than limiting, and it is acknowledged that there is no intention to exclude any future equivalents or parts thereof that are illustrated and described using such terms and expressions, and that various modifications are possible within the scope of the claimed invention.

[0256] Accordingly, although the present invention has been specifically disclosed by preferred embodiments and optional features, it should be understood that modifications and variations of the invention disclosed herein may be made by a person skilled in the art, and such modifications and variations are deemed to be within the scope of the invention disclosed herein. The invention has been described broadly and generally herein. Each of the narrower species and subgenus groups falling within the scope of the general disclosure also forms part of the invention. This includes a general description of each invention with any disclaimer or negative limitation removing any object from that category, regardless of whether the excluded material was clearly present within that category.

[0257] In addition, where features or aspects of the invention are described in the view of the Markush Group, a person skilled in the art will recognize that the invention is thereby described in the view of any individual member of the Markush Group or a subgroup of its members. Furthermore, it should be understood that the foregoing description is illustrative and not restrictive. Many embodiments will be apparent to a person skilled in the art upon reviewing the foregoing description. Accordingly, the scope of the invention should be determined without reference to the foregoing description, but instead by reference to the appended claims and the entire scope of equivalents to which such claims are applied. The disclosures of all papers and references, including patent publications, are incorporated herein by reference.

Claims

Claim 1 Compound of the following chemical formula I or its derivatives and / or its pharmaceutically acceptable salts. I Claim 2 In claim 1, a compound according to the following chemical formula. Claim 3 A pharmaceutical composition comprising the compound of claim 1 or 2 and a pharmaceutically acceptable excipient. Claim 4 A pharmaceutical composition according to paragraph 3, wherein the amount (or concentration) is sufficient to inhibit neurodegeneration in a subject, and the said amount (or concentration) sufficient to inhibit neurodegeneration is sufficient to protect retinal neurons, protect retinal tissue, protect photoreceptor cells, protect against or reduce the accumulation of autofluorescent extracellular debris, and protect against neuroinflammation. Claim 5 A pharmaceutical composition according to claim 3 or 4, wherein the compound of formula I is present in an amount of about 0.3 μM to about 20 μM or less, in some embodiments about 2 μM to about 12 μM or less, in some embodiments about 3 μM to about 10 μM or less, in some embodiments about 3 μM to about 15 μM or less, or in some embodiments about 10 μM to about 15 μM or less. Claim 6 A pharmaceutical composition according to any one of claims 3 to 5, wherein the pharmaceutically acceptable excipient comprises a sterile buffer. Claim 7 A pharmaceutical composition according to claim 6, wherein the sterile buffer is PBS buffer or saline solution. Claim 8 A pharmaceutical composition according to claim 6 or 7, wherein the sterilization buffer comprises a polysorbate-type nonionic surfactant and sucrose. Claim 9 In claim 8, the above-mentioned polysorbate-type nonionic surfactant is polysorbate 20, a pharmaceutical composition. Claim 10 A pharmaceutical composition according to any one of claims 3 to 9, comprising a delivery carrier comprising a cellulose polymer and an anionic polysaccharide, and nanoparticles comprising an amphiphilic non-ionic block copolymer and a compound of Formula I of claim 1 or 2, wherein the composition has a gelling point of about 30 °C to about 37 °C. Claim 11 A pharmaceutical composition for topical administration, in any one of paragraphs 3 to 10. Claim 12 A pharmaceutical composition for direct administration to the eye, in any one of paragraphs 3 to 11. Claim 13 A pharmaceutical composition for direct administration to the back of the eye, in any one of claims 3 to 12. Claim 14 A pharmaceutical composition that is an intravitreal injection in any one of paragraphs 3 to 13. Claim 15 A pharmaceutical composition that is an ophthalmic pharmaceutical composition in any one of paragraphs 3 to 13. Claim 16 A pharmaceutical composition according to any one of paragraphs 3 to 15, administered every other month, every month, every week, every other day, every day, twice a day, or three times a day. Claim 17 A method for protecting a neuron from neurodegeneration or treating neurodegeneration in a neuron, comprising the step of 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 requiring such contact. Claim 18 A method for treating age-related macular degeneration, comprising the step of 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. Claim 19 A method for improving retinal pigment epithelial integrity, comprising the step of administering a compound according to claim 1 or 2 or a pharmaceutical composition according to any one of claims 3 to 16 to a subject in need. Claim 20 A method according to any one of claims 17 to 19, wherein the contact or administration is made to the eye of the object requiring the contact. Claim 21 A 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. Claim 22 A method comprising the step of simultaneously administering or contacting the compound or pharmaceutical composition for the treatment of age-related macular degeneration and an additional activator to the neuron in any one of claims 17 to 21. Claim 23 A method comprising the step of sequentially administering or contacting the compound or pharmaceutical composition and additional activator for the treatment of age-related macular degeneration to the neuron in any one of claims 17 to 22. Claim 24 A method according to any one of paragraphs 17 to 23, wherein the required object is a mammal. Claim 25 A method according to any one of claims 17 to 24, further comprising the step of anesthetizing the subject requiring the contact or administration prior to the contact or administration. Claim 26 A method according to any one of claims 17 to 25, further comprising the step of dilating the pupil(s) of the subject requiring contact or administration prior to the contact or administration, wherein, optionally for claim 25, the dilation step is performed prior to or simultaneously with the anesthesia step. Claim 27 A method according to any one of claims 17 to 26, wherein the method is for protecting a neuron from neurodegeneration and the neuron is photodamaged. Claim 28 A method according to any one of claims 17 to 27, wherein the amount administered is sufficient to inhibit neurodegeneration. Claim 29 A 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. Claim 30 A method for improving retinal pigment epithelial integrity and / or reducing autofluorescent deposits in any one of claims 19 to 26. Claim 31 A method according to paragraph 30, wherein an amount sufficient to improve retinal pigment epithelial integrity and / or reduce autofluorescent deposits is administered.

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