Cannabinoid compositions and methods of using
A technology of cannabinoids, compositions, applied in the field of cannabinoid composition and usage, capable of addressing issues such as the ability to act as anti-inflammatory or immunosuppressive agents that have not been investigated by studies
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example 1
[0197] Example 1: DPPH Antioxidant Analysis
[0198] The 2,2-diphenyl-1-picrylhydrazine (DPPH) antioxidant assay was used to assess the ability of cannabinoids to act as potential antioxidants. The cannabinoids CBDA, CBD, CBDVA, CBDV, CBG, THCV and 1:1CBD:CBDA mixture had the highest antioxidant activity respectively, among which 38.19%, 48.00%, 43.81%, 44.21%, 44.28%, 45.71% and 43.74% Free radical scavenging activity.
[0199] 2,2-Diphenyl-1-picrylhydrazine (DPPH) radical (Alfa Aesar) was prepared in ethanol at 0.1 mM. Cannabinoids were tested at a final concentration of 100 uM. 2 microliters of a 7.5 mM stock solution of the cannabinoid to be tested in DMSO was added to 148 microliters of 0.1 mM DPPH to achieve a final concentration of 100 μM in the assay. Ascorbic acid (Thermo Scientific) at a final concentration of 10 mg / ml was used as a positive control. The reaction mixture was incubated at room temperature in the dark for 1 hour and the mixture was read at 517 nm u...
example 2
[0204] Example 2: LPS cytokine analysis of THP-1 human cells
[0205] One of the most robust in vitro cellular tests that can be performed to assess the immunosuppressant potential of a therapeutic agent is to determine whether the agent can suppress cytokine levels in THP-1 human monocytes that have been differentiated into macrophage-like cells using PMA And then their immune system is induced with an immunostimulant like LPS (Cochran FR, Finch-Arietta MB, 1989).
[0206] THP-1 cytokine inhibition assays were run as three independent experiments and only cytokines whose expression was induced more than two-fold in LPS control versus untreated samples were analyzed for inhibition of dexamethasone control as well as CBC, CBCA, CBD, CBDA, CBDV, CBDVA, CBG, CBGA, CBL, CBLA, CBN, CBNA, THCV, THCVA cannabinoids or a 1:1 CBD:CBDA cannabinoid blend.
[0207] THP-1 cells were maintained at 37°C in RPMI 1640 medium supplemented with 10% heat-inactivated FBS and 1% penicillin / streptom...
example 3
[0221] Example 3: Mouse LPS cytokine animal model
[0222] This experiment was performed to compare the ability of CBD, CBDA or a 1:1 equimolar mixture of CBD:CBDA to inhibit the induction of cytokines whose expression levels were induced by LPS in mice. All three cannabinoids tested were included in the analysis if two of the three cannabinoids evaluated could inhibit at least two-fold the levels of cytokines that had been induced by LPS.
[0223] Although mice have been routinely used as models of human disease, the use of mice to study inflammatory and autoimmune diseases has been particularly questioned due to the fact that the mouse immune system behaves and is regulated very differently from that of humans (Mestas and Hughes, 2004; Jameson and Masopust, 2018). However, the ability of agents to suppress the immune system can be tested in vivo using the LPS mouse model, in which mice are infused with LPS to elicit an immune response (Field et al., 1970; Ghezzi P and Spr ...
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