John Thor Arnason,

Professor, University of Ottawa

Tony Durst, John Baker

Souroubea Botanicals

Zul Merali

University of Ottawa Institute of Mental Health Research

Pablo Sanchez

Herbario JVR Universidad Nacional Costa Rica

Souroubea: A Tropical Vine that Alleviates Anxiety

Arnason Merali & Durst (uOttawa) have had a 20 year collaboration with Costa Rican

Botanists: Profs Luis Poveda, Pablo Sanchez and Marco Otarola at UNA

Genus Souroubea (Marcgraviaceae): a promising new Natural Health Product treatment for animal and human

anxiety

(Puniani et al 2015. Phytochemistry113: 73-78.)

We targeted the neotropical family

Marcgraviaceae for study:

•Previously completely unstudied

except for one published report

• Souroubea is a genus of vine species

found in Central and South America

(20 species, 6 in Central America)

•Used by Amazonian indigenous groups

for “susto”- a traditional condition in

which a fright causes disease.

•“susto” is a folk description of anxiety

•Our initial animal tests showed anti-

anxiety activity of Souroubea

“Souroubea” ethnobotany

Maya traditional healer’s Association at Indian Creek Belize:

• 4 healers treat patients with the plant

• Use leaf and bark for fever, witch craft, ulcer, dysentery.

• Dose: 20-30g/litre of water, boiled

• Drink 1 cup, 2x/day, 3days

• Adverse effects: none

• Witchcraft symptoms: patient is nervous, withdrawn, not talkative

Principal Investigators trial:

• N= 3, 2 cups of leaf tea

• Calming, no side effects, pleasant taste

Elevated Plus Maze:

Anxiety test:

rats spend much of their time exploring enclosed alleyways (safe zones) & avoid the open, ( anxiety provoking zones).

Animals treated with classical anxiolytic drugs, such as Valium, venture out more onto the open arms of the maze.

a brave little animal doing the very risky ‘unprotected head dip”

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Rats treated with of Sin-Susto crude extract (SS-CE) spent significantly more time on the open arms of the maze (as compared to controls),

Fraction F1 but not F2 has activity

Extract of Souroubea alleviated anxiety in the elevated plus-maze test (left)Isolation of active principles (right)

(Eva Puniani et al 2015 Phytochemistry)

Triterpenes in Souroubea

used in validated HPLC analysis

Co-occuring amyrins synergize

betulinic acid in unprotected head dips

Pharmacology of Anxiety Reduction:

similar to valium but with reduced side effects

such as muscle weakness and dependence.

(Mullally et al 2014, Can. J Pharm &Physiol))

FlumazeilAntagonist

Glutamate

GABA

SSA

GAD GABA-T

Presynaptic terminal

Postsynaptic neuron

GABAa

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GABA

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GAD

Kreb’s

Cycle

Glial cell

In 2010 we decided to produce Souroubea as an antianxiety

Low risk veterinary product for dogs before attempting a human product

Tony Durst (El Jefe) John Baker (Agronomist) Giovani ( grower)

Initially we collected wild plants with Mario Garcia

We needed to grow it to protect wild plants, but

Souroubea was initially difficult to cultivate or

propagate by cuttings

Then, grower Victorino Giovani used air layering

successfully at Sarapiqui site to reproduce plants

Mario Garica and John Baker planting world’s first Souroubea farm in Costa Rica 2012

Souroubea plantation 2015-16

1 harvest enough for 10 million doses

In 2013, a novel anxiolytic botanical blend, was developed by

Tony Durst: It was the final product commercialized under

trade name “Sin Susto”, now “Souroubea Botanical”

A new North American

botanical: Sycamore bark is a

rich source of betulinic acid,

reducing cost

Our tropical vine: Souroubea spp

Protected by US and Canadian patents 2012 and international PCT patents

+

Sycamore grows in Southern Ontario & eastern USAPlatanus occidentalis L. (Platanaceae)

At moderately low doses Sycamore and Souroubea are synergistic

Toxicity testing showed no effects at 10x dose

Tested as noise aversion therapy in

dogs using thunderstorm model

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Each of the treatment groups (0.5x, 1x, 2x, 4x and Diazepam) were compared with the placebo (0x) during thunder exposure:

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Inactivity Data showing canine anxiety

(hiding, freezing etc)

Treatment LSMean (sec) p value

Placebo 114.48

0.5x 106.48 NS

1x 72.61 0.01

2x 70.14 0.008

4x 90.41 NS

Diazepam 61.31 0.002

Dose dependent Stress hormone (cortisol) reduction by sin susto

in dog trial in thunderstorm model

Blend was formulated as

a chewable 3.5 g tablet for dogs

and test marketed in Canada

Onset trial: Does it work within 30 or 60 min before thunderstorm?

June 2015

Souroubea Botanicals was Incorporated

and product is now for sale on US market

From left to rightPresident Tony Durst, Zul Merali, and John Arnason

Next steps

• Humans-Post Traumatic Stress Disorder

Hope this made you feel relaxed!

Students PDFs: Eva Puniani, Ana Carballo, Amanda Sakaily, Marco Rocha,Martha Mullally, Christian Cayer, Rui liu, Kim Mitchell, Kari Kramp,

Collaborators: Mario Garcia, Pablo Sanchez Luis Poveda Marco Otarola, John Baker,Calum MacRae

Funding: NSERC, Bioniche, OCESouroubea Botanicals IncProducts from Biodiversty, Productos de la biodiversidad1956 Sharel Dr, Ottawa ON Canada K1H 6W2

Norantea guinensis Aubl.

We targeted a rare and beautiful family of neotropical vines, the MarcgraviaceaeUse reports from Amazonia suggested family use for “fear in the night” We investigated potential of Central American species as antianxiety agent

Souroubea gilgii = Sin Susto

Crossing the valley of death from bench to commercialization

Souroubea acts at the benzodiazepine receptor

SCE = supercritical extract of Souroubea, diaz=diazepam

Flu= flumazeil the GABAA antagonist

Treatment

Veh Diaz SCE Flu Flu + SCE

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