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Compositions and methods for use of cannabinoids for neuroprotection

A technology for neuroprotection and composition, which can be used in drug combinations, nervous system diseases, nervous system cells, etc., and can solve problems such as excitotoxicity and no neuroprotective effect.

Pending Publication Date: 2022-02-01
因美制药公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite the existence of anecdotal reports of cannabinoid therapeutic effects, many cannabinoids and their derivatives have been shown to exhibit no detectable neuroprotective effects at physiological concentrations
In addition, some cannabinoids and their derivatives have been shown to cause excitotoxicity at physiological concentrations

Method used

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  • Compositions and methods for use of cannabinoids for neuroprotection
  • Compositions and methods for use of cannabinoids for neuroprotection
  • Compositions and methods for use of cannabinoids for neuroprotection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] Example 1: Neuronal Cell Protection with Cannabidiol at Atmospheric Pressure

[0148] Cell Culture and Differentiation: at 37°C in 5% CO 2 The mouse 661W (RGC-5) cell line was maintained in DMEM cell culture medium supplemented with 10% FBS and 1% antibiotic-antimycotic penicillin / streptomycin (growth medium) in a humidified atmosphere. To induce neuronal differentiation in 661W cells, the medium was replaced with growth medium containing 321 nM staurosporine (STSR) and the cells were incubated at 37 °C in 5% CO 2 Incubate for 24 hours in a humidified atmosphere.

[0149] Compounds and formulations for administration: Selected Cannabinoids: CBN, CBD, CBGA, CBDA, CBND and Delta 9 -THC was purchased from Cayman and Toronto Research Chemicals. Ethanol was used as solvent to prepare 1 and 10 mM stock solutions. The treatment concentrations of prepared CBN were 0.015, 0.05, 0.15, 0.5, 1.5, 5, 10 and 15 μM. 0.5, 1.5 and 5 [mu]M treatment concentrations of other can...

Embodiment 2

[0152] Example 2: Neuroprotection of 661W cells by cannabinol under high hydrostatic pressure

[0153] All procedures were carried out as described in Example 1 unless otherwise specified. Differentiated 661W cells treated with corresponding concentrations of cannabinoids were placed in a pressurized chamber where a high hydrostatic pressure of 20-40 mmHg was maintained for 72 hours. At the end of the incubation, 661W cells were processed for MTT assay to determine cytotoxicity. result in Figure 2-Figure 5 shown in .

[0154] In a separate study, concentrations of 0.015, 0.05, 0.15, 0.5, 1.5, 5, 10 and 15 μM of the cannabinol derivative CBNA were placed on differentiated 661W cells in a pressurized chamber in which Maintain high hydrostatic pressure of about 10-25mmHg for 72 hours. At the end of the incubation, 661W cells were processed for MTT assay to determine cytotoxicity. result in Figure 6 shown in .

Embodiment 3

[0155] Example 3: Detection of cannabinoid neuroprotection in 661W cells using an apoptosis assay

[0156] Using Cell-APO Percentage TM Apoptosis is assessed with the Apoptosis Kit, which detects and measures apoptosis by a colorimetric method. The assay uses the Cell-APO Percentage TM Apoptosis system to monitor the occurrence of anchorage-dependent apoptosis in mammalian cells in vitro. It measures the executive phase of apoptosis associated with the translocation of phosphatidylserine from the inner to outer surface of mammalian cell membranes, which is experimentally supported by the binding of annexin V to phosphatidylserine. Transmembrane movement of phosphatidylserine leads to APOPercentage dye uptake by apoptotically committed cells. This dye uptake continues until blebbing occurs and is selectively imported by cells undergoing apoptosis. Necrotic cells cannot retain the dye and therefore will not be stained.

[0157] Apoptosis was assessed in differentiated 661...

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Abstract

Provided herein are methods and compositions for neuroprotection. The neuroprotective composition can be or include cannabinol or a derivative thereof. The neuroprotective composition can be used in the treatment of a neurodegenerative disease. The neuroprotective composition can be used to protect retinal neurons from degeneration in a subject in need thereof, such as for treatment of glaucoma.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 838,216, filed April 24, 2019, the contents of which are hereby incorporated by reference in their entirety for all purposes. Background technique [0003] Neurodegeneration is an underlying symptom of many different diseases of the central and peripheral nervous systems. Neurodegeneration includes neuronal atrophy, axonal degeneration (eg, Wallerian and / or Wallerian degeneration), and induction of necrotic or programmed cell death mechanisms. Different types of programmed cell death, such as apoptosis, autophagy, pyroptosis and neoplastic disease have been demonstrated in neurons. Stimuli such as physical injury, oxidative stress, excitotoxicity, mitochondrial dysfunction, inflammation, iron accumulation, and protein aggregation have all been shown to contribute to the mechanisms of neurodegeneration. [0004] Glaucoma is a form of optic nerve ...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61K9/10A61K9/107A61P25/28A61P27/02C07D311/58C07D311/60C12N5/079
CPCA61K31/352A61P25/28A61P27/02C07D311/80A61K9/0048A61K9/0019A61K47/38A61K9/1075A61P27/06
Inventor E·徐U·库马R·K·索姆万施S·邹
Owner 因美制药公司
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