Our Science

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Our Science

Equine15 NATURAL is our product specially formulated for horses.

After our initial testing for biological activities in vitro and in vivo (see below), Natural MA Inc. decided to develop a formulation for horses and explore whether it may help relieve issues with mobility observed in older horses.  The rationale to do so was twofold:  first, the product showed very promising antiviral activity in the laboratory including for Human Herpes Simplex Virus I.  There is mounting evidence that equine herpes virus infections are underdiagnosed in horses showing issues with mobility.  Second, we had encouraging results using a canine formulation of our product in ameliorating joint health in older dogs.

 

The equine formulation was notified by Health Canada and introduced in the market.  The feedback of satisfied customers is quite positive (see “Testimonials” tab) and at times moving.   The best indicator is the number of repeated customers which is about 75%.

Summary of safety considerations

The focus of this summary pertains to the safety of our product which has been demonstrated in vitro and in vivo.

 

 

Toxicity studies in vitro – University of Alberta

Our products effects were tested on human ERG potassium channel (hERG). hERG is expressed on cardiac muscle cells and is often the reason underlying toxicity of drug candidates. This assay is considered an important first screen for toxicity of novel drug candidates. Our product showed no adverse effects in all parameters tested, including activating thresholds, conductance etc. etc. This is a very important result since the vast majority of small drug candidates often fail this test.

Toxicity studies in vivo - various institutions (reported in the text)

Results using our product in several species show excellent tolerability and no toxic effects. Species included: pigs (Nutreco Canada Agresearch - 3 months, 80 mg product/kg pig); cattle (Mapleview Agri - 78 days, 40 mg/kg); dogs (CanCog Technologies – 9 weeks, 300 mg/dog - Beagles); mice (Evotec Inc. – 1 week, 200 and 400 mg/kg). Moreover, tests were conducted in salmon (U. of Maine); shrimp (Center for Aquatic Technologies).

In particular, studies in mice were conducted with the highest doses of our product tested in vivo and clinical endpoints included: posture, piloerection, respiration, salivation, tremors, convulsions, prostration, socialization, provoked behavior and change in weight.

The following parameters were also assessed: cardiac and renal effects; pulmonary odema; changes in the normal glossy coat and ulceration of the mucous membranes. All parameters were found “normal” at the end of the experiment.

In conclusion, after testing our product in different species over long periods of time, we are confident that our product is safe and well tolerated. This confidence is also supported by the approval granted by various Canadian Regulatory agencies (CFIA and Health Canada) to our product for Cattle, Horses, and Dogs.

Summary of biological activities

This summary pertains to all the tests that were performed to explore the biological activity of our product.

Tests for activities in vitro – various institutions (reported in the text)

Our product was tested in various antiviral assays performed at U. of Calgary, U. of Maine, U. of Minnesota. For all the in vitro experiments, controls were added to monitor cell toxicity. Our product was generally well tolerated even at the highest concentrations tested in human primary cells and various established cell lines.


Human Rhino Virus 16 (HRV-16) - Picornaviridae, ssRNA (+) – University of Calgary

Cellular responses of primary human lung epithelial cells to viral infection were strongly inhibited: >10 fold reduction in IP-10 release. However, viral titer in supernatants only marginally affected: 2-3 fold reduction


Infectious Salmon Anemia Virus (ISAV) - Orthomyxoviridae, ssRNA (-) – University of Maine

This virus belongs to the same viral family as the influenza virus. Viral titers in supernatants of infected ASK (Atlantic Salmon Kidney cell line) were strongly affected: 5-6 logs


Human Herpes Symplex Virus 1 (HSV-1) - Herpesviridea, dsDNA – University of Maine

A modified plaque reduction assay was performed with Vero cells. Strong inhibition at the highest dilutions tested was observed. Precise measurements of TCID50 were not possible as no visible viral CPE (cytopathic effect) was observed in any of the product concentrations used. However, it can be said that the TCID50 is >3-4 logs.


Porcine Epidemic Diarrhea Virus (PEDV) - Coronaviridae, ssRNA (+) – University of Minnesota

Studies were performed using Vero-76 cells (monkey kidney cell line): viral titer in supernatants were reduced by 1-2 logs.

Other test for activities in vivo – various institutions (reported in the text)

Dogs: naturally occurring osteoarthritis - CanCog Technologies (Toronto)

A small trial was designed to determine whether our product could help dogs suffering from joint issues. The most reliable model available was naturally occurring osteoarthritis in old beagles. Sixteen (16) dogs in good general health but with both previous radiographic evidence of osteoarthritis and behavioral evidence of osteoarthritis based on historical pain and function scores were obtained from Vivocore Inc. The subjects selected were those with the highest combined pain plus function scores. In the pain scores, 4 animals from the control group performed more poorly and 4 performed slightly better: in the treatment group, 7 performed much better and 1 more poorly (P=0.1057, chi square test). While not quite of statistical significance, these results show improvement of joint health in osteoarthritic dogs. It was proposed to repeat the experiment with Pet Dogs (vs. highly affected ones).

Testing

Conclusions

• Our product is not toxic in vitro or in vivo

• Our product displays antiviral properties in different families of viruses (panviral) in vitro

• Our product does not prevent viral infection but affects viral replication and/or viral release (attenuation)