cannabis roots: a neglected therapeutic resource? the elder, 1 st century “a decoction of the root...

Cannabis Roots: A Neglected Therapeutic Resource?

Ethan Russo, MD20402 81st Ave. SWVashon, WA 98070

[email protected]

Fuchs, New Kreuterbuch, 1542

EBR

Cannabis plant

Fan leaves: Cannabinoids (CB): 0.05%Sesquiterpenoids>>Monoterp.FlavonoidsCanniprene (up to 0.2%)

Stem:CB: 0.02%Cellulose

Roots:CB: 0%TriterpenoidsAlkaloids

Fertilized Flower:CB: up to 13%

Unfertilized Flower: CB: up to 30%Monoterp>Sesquiterp.:up to 4% total

Capitate glandular trichomes:CB: up to 60%Monoterp.>Sesquiterp.: up to 8% total

Seeds:CB: 0%Terpenoids: 0%Edestin protein: 35%Essential Fatty Acids: 35%cannabisin Bcaffeoyltyramine

Seed Sprouts:as above +cannflavin A

Russo and Marcu 2017Cannabis pharmacology: The usual suspects and afew promising leads. Adv Pharmacol (in press).

Pliny the Elder, 1st Century

“A decoction of the root in water, relaxes contractions of the joints, and cures gout and similar maladies. It is applied raw to burns, but it must be frequently changed, so as to not let it dry.”Natural History (Pliny 1951) (Book XX, Ch. 97, p.298)

Ibn Sina (Avicenna), Canon of MedicinePersia, 11th century.

Tumors and PimplesThe compress with the boiled roots of cannabis decrease fever and resolve the indurations if applied on the hot tumors and hardened places [of the body].

Leonhart Fuchs, New Kreuterbuch, 1542Hemp root, boiled in water, and wrapped---is also good for gout. The raw root, pounded and wrapped, is good for the burn.

(Translation courtesy of Franjo Grotenhermen)

FUCHS, L. 1999. The great herbal of Leonhart Fuchs : De historia stirpium commentarii insignes, 1542 , Stanford, CA, Stanford University Press.

François Rabelais, 1546“The root of this herb, boiled in water, soothes muscles, stiff joints, gout pains, and rheumatism.”RABELAIS, F. 1990. Gargantua and Pantagruel, New York, Norton. Book III, Chapter 51, page 371

Li Shih-Chen, Pen T'sao Kang Mu (Bencao Gang Mu), 1596

It was also observed, “The juice of the root is ---thought to have a beneficial action in retained placenta and post-partum hemorrhage.” Smith FP, Stuart GA. Chinese materia medica; vegetable kingdom. Shanghai,: American Presbyterian Mission Press; 1911. (p. 91)

Szymon Syrenski (Simon Syrenius), 1613

“Curved and shrunken body parts - hemp roots boiled in water.”Zielnik p. 824Translation Courtesy ofBogna Ignatowska-Jankowska

Georg Everhard Rumpf, 17th century“In Hitu [Ambon Island, Indonesia] the Moors took the root of the male or flower-bearing plant (which in European herbals are not readily distinguished) from my garden, and gave it to eat to those who were held fast by unclean Gonnorhaea;”

E.M. Beekman (Ed.). The Poison Tree: Selected Writings of Rumphius on the Natural History of the Indies. Amherst: Univ. of Massachusetts, 1981. p. 197.

Nicholas Culpeper, 17th century.“The decoction of the root allays inflammations of the head, or any other parts;” Culpeper's complete herbal. London: W. Foulsham, 1994.(p. 183)

John Parkinson, 1640.Hempe is cold and dry --- The decoction, of the roote is sayd to allay inflammations in the head or any other part, the herbe it selfe, or the distilled water thereof performeth the like effect; the same decoction of the rootes, easeth the paines of the goute, the hard tumours, or knots of the joynts, the paines and shrinking of the sinewes, and other the like paines of the hippes: it is good to be used, for any place that hath beene burnt by fire, if the fresh juyce be mixed with a little oyle or butter.

Theatrum botanicum: The theater of plants; or, An herball of a large extent. London: Tho. Cotes. (p. 598)

William Salmon, 1710XV. The Decoction of the Root. --- it is said to be good against, viz. to allay Inflammations in the Head, or any other part, to ease the pains of the Gout, to help hard Tumors or Knots in the Joints, Cramps, and shrinking of the Sinews, and to ease the pains of the Hip, or Sciatica, being applied thereto by Fomentation, and afterwards mixed applied made up into a Cataplasm with Barley Flower, renewing of it every day.

SALMON, W. 1710. Botanologia. The English herbal: or, History of Plants., London, I. Dawkes. (p. 510)

Robert James, 1747“The Root boil’d, and applied by Way of Cataplasm, mitigates Inflammations, discusses Tumors, and dissolves tophaceousConcretions at the Joints.”Book III, p. 274

Marcandier 1758“Its root, boiled in water, and coated in the form of a cataplasm, mollifies and softens the joints of the fingers that are shrunken. It is quite good against the gout, and other inflammations; it resolves tumors and callosities of the joints. Pounded and ground fresh, with butter in a mortar, one applies it to burns, which it soothes infinitely, provided it is often renewed. Its juice and decoction placed in the buttocks [anus] of horses, in fact also brings out the vermin.”(translation EBR, p. 40-41)

Makhzan al-adwiya, 18th Century“a poultice of the boiled root and leaves for discussing inflammations, and cure of erysipelas, and for allaying neuralgic pains.”as provided by: O'Shaughnessy, W. B. (1838-1840). On the preparations of the Indian hemp, or gunjah (Cannabis indica). Transactions of the Medical and Physical Society of Bengal, 71-102, 421-461.

Lyster H. Dewey, 1914

Dewey, L. H. (1914). Hemp. Yearbook of the United States Department of Agriculture 1913.Washington, DC: US Government Printing Office.

Hemp Taproots- up to 2.5 m. length

Réunion, Indian Ocean“Boiled roots were used to reduce infants’ vomiting---.”

Benoist, J. 1975. Réunion: Cannabis in a pluricultural and polyethnic society. In Cannabis and culture., edited by V. Rubin. The Hague: Mouton Publishers. (p. 230)

China“Ma gen, Cannabis Radix, cannabis (hemp) root: This is the root of the cannabis plant. Ma gen dispels stasis and stanches bleeding. It is used in the treatment of strangury, flooding and spotting, vaginal discharge, difficult delivery, retention of the placenta, and knocks and falls. It is taken orally, either as a decoction or crushed to extract its juice (in its fresh form).”(p. 137)Brand E, Wiseman N. Concise Chinese Materia Medica. Taos, New Mexico: Paradigm Publications; 2008.

Korean TeaMa cha (마차, 麻茶 or 산약차) - made from the root of hemp.

http://en.wikipedia.org/wiki/Korean_tea

A Case of Mistaken Identity• Rodger (1971) recollected from

1931 his physician uncle using “Indian hemp” roots used to treat dropsy (edema)

• This prompted an investigation by ten Ham (1973), who infused cannabis roots in whiskey into guinea pig hearts with resultant bradycardia he attributed to potassium content

• Mole (1974) tested similarly, seeing minimal bradycardia, increased PR interval and QRS amplitude, not due to potassium

• No cardiac activity of cannabis roots were noted by the ancients

“Indian hemp” in 1939 was the common name applied to Apocynum cannabinum, a known cardiotoxin, used to treat dropsy in folk medicine (Kloss, p. 346), and among the Meskwaki Native Americans of the Midwest (Smith 1928, Moerman 1998). Rodger’s report is likely spurious.

Chile“Hemp roots have been used as a purgative.” (p. 74)

Forster E. History of hemp in Chile. Journal of the International Hemp Association. 1996;3(2):72-7.

Argentina, 2008 Indian Hemp

The root bark, provides a fairly harsh taste mainly due to the presence of tannin, is used fresh in cooking at the rate of thirty grams per liter of water, or dry, fifteen grams, for abbreviating bouts of fever in malaria. The bark should be collected in the early spring, when it is also a good tonic, successfully administered pulverized and mixed with wine for weakness and pains of the stomach. It tones at the same time the entire digestive apparatus, removes toxins and infections caused by the weakness of them.

Its same fruits [seeds] can replace the root. (p. 159-160)

Manfred, L. (2008). Siete mil recetas botanicas a base de mil trescientas plantas medicinales., Buenos Aires, Argentina, Editorial Kier S.A.

Cannabis Roots, Hydroponic, Vegetatively Propagated

Cannabis Root Components

TriterpenoidsAlkaloidsSterols et al.

EBR

Triterpenoids• Metabolites of isopentenyl pyrophosphate oligomers.• The largest group of phytochemicals in nature (>20K),

predominantly in plants, but also in algae, plant waxes• Biosynthesized in plants by cyclization of squalene (also

precursor to steroids)• Insoluble in aqueous media• Display wide spectrum anti-inflammatory, antipyretic, and

anti-carcinogenic effects with low toxicity (Bishayee 2011)• Pentacyclic triterpenoids’ anti-inflammatory activity may be

relate to inhibition of 5-LOX, iNOS, COX-2 and NF-κB activity (Patlolla 2012)

• Natural triterpenoids suppress cancer growth via interference with cell cycle regulatory proteins and their expression (Patlolla 2012)

Friedelin• Isolated 1892 by Friedel, or as

early as 1805, as “cerine”• MW: 426.38 daltons• MP: 262-265°C• Found in many plants

(Aesculus, Cannabis, Citrus, Diospyros, Quercus, Rhododendron, Vaccinium), algae, lichen, mosses, peat, coal, mineral wax and swine (Chandler 1979)

• Concentration in cannabis root, calculated from Slatkin (1971): 12.8 mg/kg

Queiroga, CL et al., 2000 Evaluation of the antiulcerogenic activity of friedelan-3beta-ol and friedelin isolated from Maytenus ilicifolia

(Celastraceae). J Ethnopharmacol, 72, 465-8.

• Studied indomethacin-induced ulcers in male Wistar rats

• Friedelin showed no activity in this assay

Ee, GC, et al., 2005. Cytotoxic activities of chemical constituents from Mesua daphnifolia.

Trop Biomed, 22, 99-102.

Friedelin was weakly active against 4 cancer cell lines

Friedelin Estrogen(-) breast Cervical T-lymphoblastic Human ovariancancer cells cancer cells leukemia cells cancer cells

Antonisamy, P. et al., 2011. Anti-inflammatory, analgesic and antipyretic effects of friedelin isolated from Azima tetracantha Lam. in mouse and rat models. J Pharm

Pharmacol, 63, 1070-7.

In adult Wistar albino rats, friedelin markedly reduced carrageenan-induced hind paw edema, persisting for 6 hours. Effects of friedelin at 40 mg/kg dose were comparable to indomethacin 10 mg/kg.



Friedelin at doses of 2 or 4 mg markedly reduced ear edema after croton oil administration to adult Wistar albino rats.



Friedelin inhibited peritoneal capillary permeability after acetic acid administration in a dose-related manner in adult Wistar albino rats.



Friedelin inhibited granuloma formation after placement of cotton pellets subcutaneously in the axilla in adult Wistar albino rats.



Friedelin significantly (p<0.05) inhibited paw swelling after Freund’s adjuvant injection in adult Wistar albino rats.



Friedelin significantly (p<0.05) reduced abdominal constrictions and stretching after acetic acid injection in adult Wistar albino rats. The effect was less on first phase (0-5 min) neurogenic pain than on second phase (20-30 min) inflammatory pain.



Friedelin showed no significant effect vs. control on pain threshold in the hot plate test in adult Wistar albino rats.



Friedelin administered orally showed significant reduction in rectal temperature (p<0.05) after yeast injection in adult Wistar albino rats. Results were comparable to the anti-pyretic effect of paracetamol (acetaminophen).



Conclusions: “The results of this study have empirically indicated that friedelin was effective in the treatment of inflammatory disease. Friedelin showed potent in-vivo anti-inflammatory, analgesic and antipyretic effects. --- Due to the remarkable biological activity of friedelin it will be appropriate to conduct further research to develop it into a medicine.” (p. 1077)

Sunil, C. et al., 2013. Antioxidant, free radical scavenging and liver protective effects of friedelin isolated from Azima

tetracantha Lam. leaves. Food Chem, 139, 860-5.

Friedelin showed reducing power in vitro, comparable to BHT and ascorbate, which was dose-related.


tetracantha Lam. leaves. Food Chem, 139, 860-5. a) DPPH radical scavenging

effect: IC50 at 21.1 mMb) Hydroxyl radical

scavenging: 50% inhibition at 19.8 mM

c) Nitric oxide radical inhibition: IC50 at 22.1 mM

d) Superoxide radical scavenging: IC50 at 21.9 mM

e) Inhibition of lipid peroxidation: IC50 at 18.1 mM

Friedelin results in 5 in vitroantioxidant assays



Friedelin 40 mg/kg pre-treatment reduced CCl4-induced LFT elevations due to hepatic damage (p<0.005), comparable to silymarin extract of Silybum marianum (milk thistle).



Friedelin 40 mg/kg pre-treatment before CCl4 administration produced highly significant increases in superoxide dismutase, catalase and glutathione peroxidase levels (p<0.005) to normal values, comparable to silymarin.



Conclusion:“Friedelin showed effective antioxidant, free radical scavenging and liver protective effects both in vitro and in vivo studies. It may be helpful in prevention or slowing the progress of various oxidative stress-related diseases.” (p. 864)

Epifriedelanol• Triterpenoid found in other

plants beyond cannabis, and in microbes (Chandler 1979)

• MW: 428.4 daltons• MP: 277-278 °C (Kim 2005)• Concentration in cannabis

root: 21.3 mg/kg (calculated from Slatkin 1971)

• Produced 100% inhibition of crown gall tumors on potato disks at 40-60 μg/disk (Kundu 2000)

Yang, HH et al., 2011. Epifriedelanol from the root bark of Ulmus davidiana inhibits cellular senescence in human

primary cells. Planta Med, 77, 441-9.• Assessed adriamycin-induced cell

senescence in human fibroblasts (HDF) and human umbilical vein endothelial cells (HUVEC)

• Epifriedelanol was especially active, and also decreased SA-β-gl activity, p53 protein and reactive oxygen species (ROS)

• “This compound [epifriedelanol] may be a promising candidate for developing dietary supplements or cosmetics to modulate tissue aging-associated diseases.” (p. 448)2=Friedelin

3=Epifriedelanol

Alkaloids in Cannabis Root• Cannabisativine isolated from

Cannabis root (Slatkin 1975, Lotter 1975, Turner 1976), MP: 167-168°C (Lotter), 165-166°C (Kuethe 2004), concentration 2.5mg/kg (calculation from Turner data) or 0.0004% (Mechoulam 1988)

• Anhydrocannabisativine isolated from cannabis roots and leaves (ElSohly 1978), at calculated concentration 0.3 mg/kg, or 0.00046% (Mechoulam 1988)

• No pharmacological information available on either substance (Mechoulam 1988): “They are present in miniscule amounts and are presumably not relevant to any cannabis biological activity.” (Personal comm. 2013)

Miscellaneous Substances from Cannabis Root

Slatkin (1975) isolated from cannabis root with methanol: sitosterol: calculated content 1.5%campestrol: calculated content 0.78%stigmasterol: calculated content 0.56%Slatkin (1971) isolated N(p-hydroxy-β-phenylethyl)-p-hydroxyl-trans-cinnamamide (MW: 283.12 daltons, MP: 252-254°C), at calculated concentration of 1.6mg/kg, with analgesic activity in the mouse tail flick test at 25, 50 and 100 mg/kg sc.Roots also contain (Slatkin 1975): choline, neurineCannabis roots contain no cannabinoids (Potter 2009).

The Big Picture

Flores-Sanchez, I. J. & Verpoorte, R. (2008). Secondary metabolism in cannabis. Phytochemistry Reviews, 7, 615-639.

Initial Observations• Ancient and folk contentions concerning

therapeutic approaches with cannabis root are borne out by modern testing indicating strong anti-inflammatory and analgesic effects. Some evidence also supports activity against cancers and as a febrifuge.

• The triterpenoids, friedelin and epifriedelanol, seem the best candidates.

• Little work has been done on root alkaloids.

Possible Research Plan• Analyze cannabis roots from Canadian hemp

cultivars and drug strain chemovars (e.g., high-THC and high-CBD) at UBC and in USA, respectively.

• Reassess concentrations of key constituents, particularly friedelin and epifriedelanol, as these are likely of greatest pharmacological import and promise.

• Consider additional testing of root alkaloids.• Consider animal testing of root extracts.• Consider therapeutic targets and vehicles.

cannabis roots: a neglected therapeutic resource? the elder, 1 st century “a decoction of the root...

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