NT-VRL™ Terpene Formulation

“…the terpene formulation combined with CBD is twice as effective at using CBD on its own…”   

Terpenes have been proven for their potent anti-inflammatory properties by inhibiting different biochemical mechanisms in the body’s inflammation process.

NT-VRL™ is a patented, data-driven terpene formulation developed and manufactured by Eybna. The formulation contains terpenes with a high indication for their anti-inflammatory properties, in optimized ratios. The formulation is based on Eybna’s proprietary phytochemical affinity database, utilizing Eybna’s advanced data-mining and formulation design methodologies, to develop an effective and safe formulation.

In order to evaluate NT-VRL™ efficacy, the formulation was tested in a validated biological in-vitro model that mimics an inflammatory response in the human body.

NT-VRL™ exhibited a strong anti inflammatory activity, and even more profoundly when combined with CBD.

As seen in:

Eybna’s Data-Driven Approach

The cannabis plant consists of a wide variety of phytochemicals working together to create its unique therapeutic qualities.

Eybna collects and studies the different phytochemicals found in cannabis and in other essential oils to learn their chemical, physical and biological properties with the goal to utilize them for human targeted consumption and different applications.

Eybna’s phytochemical database aggregates a wide variety of in-vitro models as well as in-vivo studies together with modern computer aided in-silico techniques.

Eybna Data-Driven functional formulations are designed for efficacy, safety and synergetic qualities, bringing scientific evidence to cannabis products.

The Study

Abstract

Eybna performed an in vitro immunology evaluation of a terpene formulation called NT-VRL™. The primary goal of this study was to assess the effect of the formulation on cytokine secretion levels in human immune system resembling the cytokine storm in the human body. The immunologic assay included evaluation of the effect of the test compounds (in different concentrations) on the in vitro cytotoxicity and cytokine production. Experiments were conducted on commercial human peripheral blood mononuclear cells (PBMCs) from two healthy donors.
Results of the LPS-induced cytokine assay show that NT-VRL™ had a dose-dependent inhibitory effect on the secretion of pro-inflammatory cytokines, without compromising cell viability.

Background

In the human body, inflammatory response is a main component of the immune response against pathogens. The white blood cells use a set of sensors to recognize the pathogen and produce cytokines in response. These cytokines trigger inflammation and recruit other cells of the immune system to fight the pathogen. When the production of cytokines is excessive and uncontrolled it is described as “cytokine storm”. Excessive chronic production of pro-inflammatory cytokines contributes to inflammatory diseases and different pathologies.

NT-VRL™ is a patented proprietary terpene formulation developed and manufactured by Eybna Technologies Ltd. NT-VRL™ Data-Driven formulation is composed of 30 individual terpenes that are potential anti inflammatory agents – all natural, highly purified and Generally Recognised As Safe.

Methods

A preclinical ex vivo immunological evaluation of
Eybna’s terpene formulation was performed in
collaboration with Cannasoul Analytics Ltd. to
measure its effect on innate immune response.
These assays included:

Cytotoxicity Assay

Primary human PBMCs were suspended in a suitable culture media at in a flat bottom 96-well plated and incubated with each test-item for 24 hours. Cell viability was assayed by using Alamar blue® reagent. The control compounds included: Saline & DMSO alone as non-toxic solutions and Triton (1%) as a cytotoxic reagent.

 

Cytokine Expression assay

Human PBMCs were thawed and suspended in a designated culture media in 24-well plates. The controls and test compounds were diluted in culture media to achieve the final assay concentration. Cells were incubated with the test compounds and controls for 1 hour, treated with 100ng/ml LPS and incubated for 24 hours. 1 ug/ml dexamethasone (DEX) served as positive control. Following incubation, the supernatants were collected and the levels of a panel of secreted cytokines were measured by ELISA (R&D systems) and the optical density were determined by ‘CLARIOStar-plus’ plate-reader. The concentration of cytokines was calculated using a standard curve.

The Study Results

Cytotoxicity assay

In this AlamarBlue® assay percentage of viable cells was calculated out of NT (No Treatment) negative control. The results of the cytotoxicity test are summarized in the Figure 1. These results provided the applicable working concentrations of these treatments at below 20μg/ml.

Cytokine Secretion

Results for all four cytokines (IL-1β, TNF-α, IL-6 and IL-8) show that NT-VRL™ formulation has a dose-dependent inhibitory effect on the secretion of pro-inflammatory cytokines, without compromising cell viability (Figure 2). The anti inflammatory effect of NT-VRL™ formulation could be seen in concentrations 2 – 20μg/ml. The inhibition magnitude of 20μg/ml NT-VRL™ is about 8, 17 and 7% higher than the effect of 2μg/ml CBD in regards to IL-1b, TNFa and IL-6, respectively. In addition, 20μg/ml NT-VRL™ showed cytokine

Conclusions

High levels of pro-inflammatory cytokines are in direct correlation with the disease severity, that indicated on the immune response. By reducing the cytokine storm that occurs in many inflammatory conditions, the immune system regains control of the infection. The aim of this in-vitro model is to assess Eybna’s NT-VRL™ as an anti-inflammatory agent. Taken together, the results suggest that NT-VRL™ exert a dose-dependent inhibition of LPS-induced cytokine secretion, higher than Dexamethasone. Moreover, combining CBD with Eybna’s NT-VRL™ presented the highest reduction of all cytokine secretion and resided in what appear to be an additive effect. The presented information indicates that NT-VRL™️ might be effective as anti-inflammatory agent and may potentiate other immunomodulators such as CBD.

 

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