The invention relates to the field of CBD.
CBD is believed by many to have therapeutic effects and it is known that relatively high doses of CBD decreases VTA dopamine activity. This inhibition of dopamine activity is related to the pharmacotherapeutic properties of CBD in producing anti-psychotic effects, anti-addictive effects, anti-anxiety effects and potential alleviation of traumatic memory disorders such as PTSD.
Forming one aspect of the invention is a formulation comprising CBD and a PPAR agonist.
According to another aspect, the PPAR agonist can be an omega-3 based PPAR agonist.
According to another aspect, the PPAR agonist can comprise substantially equal parts DHA and EPA.
Advantages, features and characteristics of the invention will become evident to persons of ordinary skill in the art upon review of the following description.
Forming an embodiment of the invention is a formulation comprising CBD and a PPAR agonist, the PPAR agonist being an omega-3 based PPAR agonist comprising substantially equal parts DHA and EPA.
Numerous experiments were conducted.
Electrophysiology Experiments
Three experiments were run of this type. All included a vehicle. In addition, the components were as follows:
Experiment 1:
100 ng CBD; 100 ng CBD+100 ng T007; 100 ng T007
Experiment 2:
50 ng CBD; 0.5 nmol n-3; 50 ng CBD+0.5 nmol n-3
Experiment 3:
1 ng CBD; 0.25 nmol n-3; 1 ng CBD+0.25 nmol n-3; 1 ng CBD+0.25 nmol n-3+100 ng T007
In the above:
The omega-3 formulation includes equal amounts of DHA and EPA. Two doses were used in this study. 0.5 nmol n-3 is made up of 0.5 nmol DHA and 0.5 nmol EPA. 0.25 nmol n-3 is made up of 0.25 nmol DHA and 0.25 nmol EPA
In this procedure, rats were anesthetized with urethane. An electrode was used to record the baseline frequency of dopamine neurons in the ventral tegmental area (VTA) for 5 minutes. After the baseline recording, combinations of CBD, omega-3, and T007 (a PPARg antagonist) were infused into the nucleus accumbens (NAc).
The post-infusion frequency of that neuron was compared to its pre-infusion frequencies.
Although the mechanisms are unclear, evidence suggests that CBD produces its therapeutic effects through the decrease of dopamine neuron activity in the VTA, as detailed below.
Experiment 1
One of the characteristics of dopamine cells is that they have a tonic mode of firing and a burst mode of firing. Therefore, both the frequency and burst changes following infusions were analyzed.
One-way ANOVA revealed that these groups are not significantly different.
However, it can be seen that 100 ng caused a decreasing trend in VTA dopamine frequency.
Experiment 2
In the second experiment, a lower dose of CBD (50 ng) was combined with omega-3 fatty acids.
With reference to
With reference to
Experiment 3
In this experiment, an attempt was made to replicate the results of experiment 2 with lower doses of CBD and omega-3.
With reference to
With reference to
Elevated Plus Maze Experiments
In these experiments, the following was measured:
1. Time in open arms
2. Entries into open arms
CBD produces its anxiolytic effects by decreasing VTA dopamine activity. We have just seen that CBD causes a slight decrease in VTA dopamine activity while combined CBD+omega-3 caused significant decreases.
An anxiolytic effect would therefor be expected when CBD and omega-3 are infused into the NAc of rats.
In these experiments, the apparatus of
As shown, the apparatus includes 2 open arms and 2 closed arms that form a plus shape. Rats feel more secure in closed spaces and thus it would be expected that more anxious rats would spend more time in the closed arms and less time on the open arms.
Before each test, the same doses of drugs used in the electrophysiology experiments were infused.
The rat was then placed in the middle of the apparatus, facing into the closed arms and recorded for 10 minutes.
The number of entries into the open arms and the total time spent in the open arms was recorded. Less anxious rats would be expected to make more entries into the open arms and spend more time there as well.
Experiment 1
As shown in
Experiment 2
As shown by in
While 50 ng CBD was not effective, post-hoc analysis revealed that 0.5 nmol omega-3 significantly increased the time spent in open arms and entries into open arms as well.
The combination of 50 ng CBD and 0.5 nmol omega-3 had the same effect.
To determine if CBD and omega-3 would have a synergistic effect, the dose of CBD and omega-3 was decreased in experiment 3.
Experiment 3
In
In
Light Dark Box Experiment
In these experiments, the following was measured:
1. First transition to the dark box
2. Second transition to the light box
3. Total number of transitions
4. Time spent in the light box
The light-dark box is another anxiety test.
This experiment uses the apparatus made up of 2 boxes as shown in
Rats would prefer to be in the dark box and would be anxious about going into the light box.
Following the infusion, the rate is placed in the light box facing away from the opening to the dark box. A 10 minute recording is made. During these 10 minutes, measurements are made of four things:
1. The time it takes for the rat to make the first transition into the dark box
2. The time it takes for the rat to make the second transition from the dark box back into the light box
3. Number of transitions between boxes
4. Time spent in light box
Therefore, it would be expected than less anxious rats would take a longer time to make the first transition into the dark box, spend less time to make the second transition into the light box, make more transitions between boxes, and spend more time in the light box.
Experiment 1
As shown in
As shown in
Experiment 2
As shown in
Similar to
The doses of CBD and omega-3 were decreased in experiment 3 to attempt to find a clear synergistic effect with CBD and omega-3.
Experiment 3
In
In
With reference to
Note that in
Open Field Experiments
In these experiments, the following was measured:
1. Time spent in center zone
2. Entry into center zone.
3. Total ambulatory time
4. Total ambulatory distance
Based on the results from
In the open field test, illustrated in
At the same time, we can also measure anxiety. Rats would prefer to be on the outer edges of the box so that one side of their body is covered by the wall of the box. They would feel anxious about being exposed in the center of the box. Therefore, less anxious rats would be expected to spend more time in the center zone.
Experiment 1
With reference to
With reference to
Olfactory Fear Conditioning Experiments
The olfactory fear conditioning protocol is for measurement of the formation of fear memory. This protocol lasts three days. On day 1, the rat is habituated to two boxes (one with a striped background and one with a polka dot back ground).
On day 2, the rat receives a drug infusion and is placed into one of the boxes (previously assigned as the “shock box”). While in the box, the rat is exposed to 2 odours (peppermint and almond). The rat is exposed to one odour followed by the other odour 5 times. One of these odours were previously assigned as the “shock” odour (CS+) such that after exposure to the “shock” odour, the rat would receive a foot shock. There was no foot shock following exposure to the “safe” odour (CS−). On day 3, the rat was placed in the safe box. They were exposed to each odour (both CS+ and CS−) one at a time for 5 minutes. During the 5 minutes, freezing behaviour was recorded.
It would then be expected that vehicle rats would have associated the correct CS+ odour with the footshock and therefore demonstrate increased freezing during the CS+ odour exposure on day 3 compared to CS− exposure. It would be expected that CBD would block the formation of fear memory and the rats would freeze a similar amount to CS+ and CS−.
Experiment 1
2-way repeated measures ANOVA was performed for the three experiments. With reference to
Experiment 2
In experiment 2, there was a significant interaction effect (p=0.013) by groups and CS. While the VEH had a significant difference between CS+ and CS− freezing time, the other groups did not which suggests that the other groups are blocking fear memory formation, as shown by
Experiment 3
In experiment 3, as shown in
Note in vehicle, 1 ng CBD, and 0.25 nmol omega-3, there was a significant difference between the freezing time to CS+ and CS− which suggests that those groups have working fear memory formation. The combined 1 ng CBD+0.25 nmol omega-3 dose blocks the formation of fear memory. This effect seems to be blocked slightly by T007.
Succinctly, the aforementioned experimentation reveals that the co-administration of a PPAR agonist allowed for CBD dosage to be significantly reduced while maintaining CBD's inhibitory effects on DA neuron activity states as well as potentiating CBD's ability to produce anti-anxiety and anti-PTSD effects in pre-clinical models of these disorders in rodents. Further, the combination of CBD+Omega-3 inhibited ‘bursting rates’. This is important because bursting of DA neurons is linked to DA dysregulation in mental health disorders like addiction, schizophrenia and anxiety. This suggests that the combination may be more effective than CBD alone in treating a variety of mental health disorders including addiction, anxiety, schizophrenia: basically, anything that can normalize abnormal DA firing/bursting rates is likely good for these disorders.
It will be apparent that this has significant advantage, since CBD is relatively costly. As well, high doses of CBD creates huge bolus concentrations, which creates the potential for side-effects.
Whereas in this document, specific dosage regimes are described, it will be appreciated by persons of ordinary skill that the dosages mentioned have been proven to be useful in the context of rats and in experimental conditions; routine experimentation will be required to modify the dosages for human use.
Whereas a specific PPAR agonist is described, namely, a combination of DHA and EPA in substantially equal amounts, it is contemplated that other PPARA agonists might be useful, including but not limited to: Honokiol, magnolol, Echinacea purpurea (L.), Panax ginseng and 10-hydroxy-octadecanoic acid.
Accordingly the invention should be understood to be limited only by the accompanying claims, purposively construed.
The contents of U.S. Provisional Patent Application Ser. No. 63/007,529 are incorporated herein by reference.
Filing Document | Filing Date | Country | Kind |
---|---|---|---|
PCT/CA2021/050478 | 4/9/2021 | WO |
Number | Date | Country | |
---|---|---|---|
63007529 | Apr 2020 | US |