Bay Area Cannabis

Alternatives for Healing and Causes for Disease
You are currently browsing the Cannabinoids category

Turned-Off Cannabinoid Receptor Turns on Colorectal Tumor Growth

  • October 18, 2012 4:55 pm

HOUSTON, Aug. 1, 2008 – New preclinical research shows that cannabinoid cell surface receptor CB1 plays a tumor-suppressing role in human colorectal cancer, scientists report in the Aug. 1 edition of the journal Cancer Research.

CB1 is well-established for relieving pain and nausea, elevating mood and stimulating appetite by serving as a docking station for the cannabinoid group of signaling molecules. It now may serve as a new path for cancer prevention or treatment.

“We’ve found that CB1 expression is lost in most colorectal cancers, and when that happens a cancer-promoting protein is free to inhibit cell death,” said senior author Raymond DuBois, M.D., Ph.D., provost and executive vice president of The University of Texas M. D. Anderson Cancer Center.

DuBois and collaborators from Vanderbilt-Ingram Cancer Center also show that CB1 expression can be restored with an existing drug, decitabine. They found that mice prone to developing intestinal tumors that also have functioning CB1 receptors develop fewer and smaller tumors when treated with a drug that mimics a cannabinoid receptor ligand. Ligands are molecules that function by binding to specific receptors. Agonists are synthetic molecules that mimic the action of a natural molecule.

“Potential application of cannabinoids as anti-tumor drugs is an exciting prospect, because cannabinoid agonists are being evaluated now to treat the side-effects of chemotherapy and radiation therapy,” DuBois said. “Turning CB1 back on and then treating with a cannabinoid agonist could provide a new approach to colorectal cancer treatment or prevention.”

Cannabinoids are a group of ligands that serve a variety of cell-signaling roles. Some are produced by the body internally (endocannabinoids). External cannabinoids include manmade versions and those present in plants, most famously the active ingredient in marijuana (THC).

Receptor shutdown by methylation

Endocannabinoid signaling is important to the normal functioning of the digestive system and has been shown to protect the colon against inflammation. Since chronic inflammation is a known risk factor for colorectal cancer, the researchers decided to look into the role of cannabinoid receptors in a mouse model of colon cancer.

“People have looked at cannabinoids in cancer earlier, mainly in cell culture experiments,” DuBois said. “The molecular mechanisms for loss of the receptor and its effect on cancer have not been previously shown.”

First, the team found that CB1 was largely absent in 18 of 19 human tumor specimens and in 9 of 10 colorectal cancer cell lines. Further experimentation showed that the gene that encodes the CB1 protein was not damaged, but shut down chemically by the attachment of methyl groups – a carbon atom surrounded by three hydrogen atoms – to the gene encoding CB1.

Treating cell lines with decitabine, a demethylating agent approved for some types of leukemia, removed the methyl groups, restoring gene expression in 7 of 8 cell lines and full expression of CB1 protein in three lines.

Next, the group found that deletion of the CB1 gene in a strain of mice that spontaneously develops precancerous polyps resulted in a 2.5-to-3.8-fold increase in the number of polyps and a 10-fold increase in the number of large growths, those most likely to develop into cancer.

Treating mice that had the CB1 receptor with an endocannabinoid agonist resulted in a decline in polyps ranging from 16.7 percent to 50 percent. The reduction was greater for larger polyps.

CB1 thwarts survivin, a protein that protects cancer

Cannabinoids previously had been shown to kill cancer cells in lab experiments by inducing apoptosis – programmed cell death. The team confirmed the role of CB1 in apoptosis, showing that tumor cells with high CB1 expression were sensitive to apoptosis when treated by a cannabinoid agonist. Cell lines with silenced CB1 resisted cell death.

A series of experiments showed that CB1 increases cancer cell death by stifling a protein called survivin. Survivin is overexpressed in nearly every human tumor but is barely detectable in normal tissue, DuBois noted. Overexpression of survivin is associated with poor outcome and reduced apoptosis in colorectal cancer patients. The researchers pinpointed a cell signaling pathway by which activated CB1 cuts down survivin.

“Just increasing the levels of cannabinoids to treat colorectal cancer won’t work if the CB1 receptor is not present,” DuBois said. This suggests that treating first with a demethylating agent, such as decitabine, to reactivate CB1 in the tumor and following up with a cannabinoid might be an effective attack on colorectal cancer.

Scarcity of CB1 also is associated with Huntington’s disease, Alzheimer’s disease and multiple sclerosis. Further investigation, the researchers note, is needed to define its role in those diseases and other types of cancer. The team also analyzed the other main cannabinoid receptor, CB2, and found no role for it in colorectal cancer.

They also treated the mice with a CB1 antagonist, a compound that binds to the receptor but does not activate it. Mice with CB1 blocked in this manner also showed an increase in the number and size of polyps. A CB1 antagonist called rimonabant is currently marketed overseas for weight loss. The researchers note that a patient’s risk for colorectal cancer should be assessed when use of such drugs is being considered.

The study was funded by grants from the National Cancer Institute and the National Colorectal Cancer Research Alliance.

Co-authors with DuBois are first author Dingzhi Wang, Ph.D., Haibin Wang, Ph.D., Wei Ning, Michael Backlund, Ph.D., and Dushansu K. Dey, Ph.D., all of the Vanderbilt-Ingram Cancer Center.

Medical marijuana uses – 700 medical marijuana clinical studies and papers

  • September 26, 2012 4:09 pm

700 uses of Medical Marijuana | Sorted by Disease | ADD – Wilson’s Disease | Links to 700 Clinical Studies | Medical Marijuana Reference | Cannabis as Medicine

Medical marijuana and cannabis studies A collection of clinical studies, papers and reference providing the ultimate resource for medical disorders helped by medical marijuana.

 

 

ADD/ ADHD
Marijuana and ADD Therapeutic uses of Medical Marijuana in the treatment of ADD
http://www.onlinepot.org/medical/add&mmj.htm

Cannabis as a medical treatment for attention deficit disorder
http://www.chanvre-info.ch/info/en/…-treatment.html

Cannabinoids effective in animal model of hyperactivity disorder
http://www.cannabis-med.org/english…el.php?id=162#4

Cannabis ‘Scrips to Calm Kids?
http://www.foxnews.com/story/0,2933,117541,00.html

Addiction risk- Physical
Women’s Guide to the UofC
http://wguide.uchicago.edu/9substance.html

Cannabis Basics
http://www.erowid.org/plants/cannab…is_basics.shtml

10 Things Every Parent, Teenager & Teacher Should Know About Marijuana (4th Q)
http://www.erowid.org/plants/cannab…is_flyer1.shtml

Marijuana Myths, Claim No. 9
http://www.erowid.org/plants/cannab…bis_myth9.shtml

AIDS – see HIV

Alcoholism
Role of cannabinoid receptors in alcohol abuse
http://www.medicalnewstoday.com/articles/30338.php

Cannabidiol, Antioxidants, and Diuretics in Reversing Binge Ethanol-Induced Neurotoxicity
http://jpet.aspetjournals.org/cgi/c…ourcetype=HWCIT

Cannabis substitution
http://www.cannabis-med.org/studies…how.php?s_id=86

Cannabis as a Substitute for Alcohol
http://ccrmg.org/journal/03sum/substitutealcohol.html

ALS
Cannabinol delays symptom onset
http://www.ncbi.nlm.nih.gov/sites/e…t_uids=16183560

Marijuana in the management of amyotrophic lateral sclerosis
http://www.medscape.com/medline/abstract/11467101

Cannabis use in patients with amyotrophic lateral sclerosis.
http://www.medscape.com/medline/abstract/15055508

Cannabis Relieves Lou Gehrigs Symptoms
http://www.rense.com/general51/lou.htm

Cannabis’ Potential Exciting Researchers in Treatment of ALS, Parkinson’s Disease
http://66.218.69.11/search/cache?ei…&icp=1&.intl=us

Alzheimers
MARIJUANA SLOWS ALZHEIMER’S DECLINE
http://www.mapinc.org/drugnews/v05/n307/a10.html

Marijuana may block Alzheimer’s
http://news.bbc.co.uk/2/hi/health/4286435.stm

Prevention of Alzheimer’s Disease Pathology by Cannabinoids
http://www.jneurosci.org/cgi/content/abstract/25/8/1904

Marijuana’s Active Ingredient Shown to Inhibit Primary Marker of Alzheimer’s Disease
http://www.pacifier.com/~alive/articles/ca060809.htm

Dronabinol in the treatment of agitation in patients with Alzheimer’s disease with anorexia
http://www.cannabis-med.org/studies…how.php?s_id=61

Dronabinol in the treatment of refractory agitation in Alzheimer’s disease
http://www.cannabis-med.org/studies…how.php?s_id=92

Effects of dronabinol on anorexia and disturbed behavior in patients with Alzheimer’s disease.
http://www.cannabis-med.org/studies…how.php?s_id=59

Cannabinoids reduce the progression of Alzheimer’s disease in animals
http://www.cannabis-med.org/english…el.php?id=187#1

Molecular Link between the Active Component of Marijuana and Alzheimer’s Disease Pathology
http://www.unboundmedicine.com/medl…sease_Pathology

THC inhibits primary marker of Alzheimer’s disease
http://www.cannabis-med.org/english…el.php?id=225#3

——— Page 1

Amotivational Syndrome
Amotivational Syndrome
http://leda.lycaeum.org/?ID=12454

Marijuana Myths, Claim No. 11
http://www.erowid.org/plants/cannab…is_myth11.shtml

Debunking ‘Amotivational Syndrome’
http://www.mapinc.org/drugnews/v06/n400/a06.html

Amotivational Syndrome
http://www.bookrags.com/Amotivational_syndrome

Debunking the Amotivational Syndrome
http://www.drugscience.org/Petition/C3F.html

Cannabis Use Not Linked To So-Called “Amotivational Syndrome”
http://www.norml.org/index.cfm?Grou…tm_format=print

Anecdotal Evidence/First person stories
Shared Comments and Observations
http://www.rxmarihuana.com/comments…bservations.htm

Cannabis Sativa (Marijuana) for Fibromyalgia
http://www.fibromyalgia-reviews.com/Drg_Marijuana.cfm

ANECDOTAL ARTICLES
http://cannabislink.ca/medical/#medanecdotal

Testimonials
http://www.benefitsofmarijuana.com/testimonials.html

Excerpts of testimonials.
http://www.ganjaland.com/freemedicalseeds.htm

Appetite Stimulant
Dronabinol an effective appetite stimulant?
http://www.cannabis-med.org/studies…ow.php?s_id=188

THC improves appetite and reverses weight loss in AIDS patients
http://www.cannabis-med.org/studies…ow.php?s_id=189

Efficacy of dronabinol alone and in combination
http://www.cannabis-med.org/studies…ow.php?s_id=191

Dronabinol and marijuana in HIV-positive marijuana smokers: caloric intake, mood, and sleep.
http://www.cannabis-med.org/studies…ow.php?s_id=190

The synthetic cannabinoid nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases
http://www.cannabis-med.org/studies…ow.php?s_id=180

Safety and efficacy of dronabinol in the treatment of agitation in patients with Alzheimer’s disease
http://www.cannabis-med.org/studies…how.php?s_id=61

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Effects of dronabinol on anorexia and disturbed behavior in patients with Alzheimer’s disease
http://www.cannabis-med.org/studies…how.php?s_id=59

Dronabinol as a treatment for anorexia associated with weight loss in patients with AIDS.
http://www.cannabis-med.org/studies…how.php?s_id=21

Delta-9-tetrahydrocannabinol for appetite stimulation in cancer-associated anorexia
http://www.cannabis-med.org/studies…how.php?s_id=52

Effect of dronabinol on nutritional status in HIV infection.
http://www.cannabis-med.org/studies…ow.php?s_id=150

Dronabinol stimulates appetite and causes weight gain in HIV patients.
http://www.cannabis-med.org/studies…how.php?s_id=20

Dronabinol effects on weight in patients with HIV infection.
http://www.cannabis-med.org/studies…how.php?s_id=45

Recent clinical experience with dronabinol.
http://www.cannabis-med.org/studies…how.php?s_id=90

Dronabinol enhancement of appetite in cancer patients.
http://www.cannabis-med.org/studies…ow.php?s_id=149

Effects of smoked marijuana on food intake and body weight
http://www.cannabis-med.org/studies…ow.php?s_id=117

Behavioral analysis of marijuana effects on food intake in humans.
http://www.cannabis-med.org/studies…ow.php?s_id=118

Cancer-related anorexia-cachexia syndrome
http://www.unboundmedicine.com/medl…xia_Study_Group

THC effective in appetite and weight loss in severe lung disease (COPD)
http://www.cannabis-med.org/english…el.php?id=191#2

Machinery Of The ‘Marijuana Munchies’
http://www.sciencedaily.com/release…51226102503.htm

Arthritis
Cannabidiol is an oral anti-arthritic therapeutic in murine collagen-induced arthritis
http://www.pnas.org/cgi/content/full/97/17/9561

The Cannabinergic System as a Target for Anti-inflammatory Therapies
http://www.ingentaconnect.com/conte…000013/art00008

Sativex in the treatment of pain caused by rheumatoid arthritis
http://rheumatology.oxfordjournals….bstract/45/1/50

Suppression of fibroblast metalloproteinases by ajulemic acid,
http://ccicnewsletter.com/index.php…06_Rheumatology

The antinociceptive effect of Delta9-tetrahydrocannabinol in the arthritic rat
http://www.unboundmedicine.com/medl…binoid_receptor

Synergy between Delta(9)-tetrahydrocannabinol and morphine in the arthritic rat
http://www.unboundmedicine.com/medl…e_arthritic_rat

Cannabis based medicine eases pain and suppresses disease
http://www.medicalnewstoday.com/articles/33376.php

Pot-Based Drug Promising for Arthritis
http://www.webmd.com/rheumatoid-art…g-for-arthritis

Asthma
The Cannabinergic System as a Target for Anti-inflammatory Therapies
http://www.ingentaconnect.com/conte…000013/art00008

Acute and subacute bronchial effects of oral cannabinoids.
http://www.cannabis-med.org/studies…how.php?s_id=44

Comparison of bronchial effects of nabilone and terbutaline
http://www.cannabis-med.org/studies…how.php?s_id=43

Bronchial effects of aerosolized delta 9-tetrahydrocannabinol
http://www.cannabis-med.org/studies…ow.php?s_id=109

Bronchodilator effect of delta1-tetrahydrocannabinol administered by aerosol
http://www.cannabis-med.org/studies…how.php?s_id=60

Effects of smoked marijuana in experimentally induced asthma.
http://www.cannabis-med.org/studies…how.php?s_id=57

Marijuana and oral delta9-tetrahydrocannabinol on specific airway conductance
http://www.cannabis-med.org/studies…how.php?s_id=67

New Synthetic Delta-9-THC Inhaler Offers Safe, Rapid Delivery
http://www.medicalnewstoday.com/articles/22937.php

Smoked marijuana and oral delta-9-THC on specific airway conductance in asthmatic subjects
http://www.ukcia.org/research/Smoke…InAsthmatic.php

Atherosclerosis

Marijuana Chemical Fights Hardened Arteries
http://www.webmd.com/heart-disease/…rdened-arteries

Does Cannabis Hold the Key to Treating Cardiometabolic Disease
http://www.medscape.com/viewarticle/525040_print

Cannabis may keep arteries clear
http://www.gnn.tv/headlines/2634/Ca…_arteries_clear

The Cannabinergic System as a Target for Anti-inflammatory Therapies
http://www.ingentaconnect.com/conte…000013/art00008

Cannabis compound tackles blood vessel disease
http://www.medicalnewstoday.com/articles/22658.php

Medical marijuana: study shows that THC slows atherosclerosis
http://thenexthurrah.typepad.com/th…al_marijua.html

Cardiovascular Effects of Cannabis
http://www.idmu.co.uk/canncardio.htm

Atrophie Blanche
Atrophie Blanche Treated With Cannabis and/or THC
http://ccrmg.org/journal/04spr/clinical.html#thm

Autism
Autism and Medical Marijuana
http://www.autism.org/marijuana.html

THE SAM PROJECT: James D.
http://www.letfreedomgrow.com/articles/james_d.htm

Medical marijuana: a valuable treatment for autism?
http://www.autismwebsite.com/ari/ne…r/marijuana.htm

——— Page 2

Cancer – breast
Anandamide inhibits human breast cancer cell proliferation
http://www.pnas.org/cgi/content/abstract/95/14/8375

Inhibition of Human Breast and Prostate Cancer Cell Proliferation1
http://endo.endojournals.org/cgi/co…tract/141/1/118

Antitumor Activity of Plant Cannabinoids
http://jpet.aspetjournals.org/cgi/c…ract/318/3/1375

9-Tetrahydrocannabinol Inhibits Cell Cycle Progression in Human Breast Cancer
http://cancerres.aacrjournals.org/c…ract/66/13/6615

Cannabidiol inhibits tumour growth in leukaemia and breast cancer
http://www.cannabis-med.org/english…el.php?id=220#2

THC and prochlorperazine effective in reducing vomiting in women following breast surgery
http://www.cannabis-med.org/english…el.php?id=219#1

Cancer- colorectal
Anandamide, induces cell death in colorectal carcinoma cells
http://gut.bmj.com/cgi/content/abstract/54/12/1741

Cannabinoids and cancer: potential for colorectal cancer therapy.
http://www.medscape.com/medline/abstract/16042581

Cancer- glioma/ brain
Anti-tumor effects of cannabidiol
http://www.hempworld.com/HempPharm/…milanstudy.html

Pot’s cancer healing properties
http://www.november.org/stayinfo/br…ncerKiller.html

Cannabinoids Inhibit the Vascular Endothelial Growth Factor Pathway in Gliomas
http://cancerres.aacrjournals.org/c…hort/64/16/5617

Inhibition of Glioma Growth in Vivo
http://cancerres.aacrjournals.org/c…/61/15/5784.pdf

Delta(9)-tetrahydrocannabinol in patients with recurrent glioblastoma multiforme.
http://www.cannabis-med.org/studies…ow.php?s_id=193

Cannabidiol triggers caspase activation and oxidative stress in human glioma cells.
http://www.ihop-net.org/UniPub/iHOP…l?pmid=16909207

Cannabinoid receptors in human astroglial tumors
http://www.brainlife.org/abstracts/…t_j20060800.pdf

Cannabis extract makes brain tumors shrink, halts growth of blood vessels
http://www.medicalnewstoday.com/articles/12088.php

THC tested against brain tumour in pilot clinical study
http://www.cannabis-med.org/english…el.php?id=222#1

Cancer- leukemia
Cannabis-induced cytotoxicity in leukemic cell lines
http://bloodjournal.hematologylibra…ract/105/3/1214

Cannabidiol-Induced Apoptosis in Human Leukemia Cells
http://molpharm.aspetjournals.org/c…stract/70/3/897

Marijuana’s Active Ingredient Kills Leukemia Cells
http://www.treatingyourself.com/vbu…read.php?t=7107

Targeting CB2 cannabinoid receptors to treat malignant lymphoblastic disease
http://bloodjournal.hematologylibra…t/100/2/627.pdf

Cannabinoids induce incomplete maturation of cultured human leukemia cells
http://www.osti.gov/energycitations…osti_id=5164483

{Delta}9-Tetrahydrocannabinol-Induced Apoptosis in Jurkat Leukemia T Cells
http://mcr.aacrjournals.org/cgi/con…bstract/4/8/549

Cannabidiol inhibits tumour growth in leukaemia and breast cancer
http://www.cannabis-med.org/english…el.php?id=220#2

Cancer- lung
Antineoplastic activity of cannabinoids
http://www.ukcia.org/research/Antin…ds/default.html

Delta(9)-Tetrahydrocannabinol inhibits epithelial growth factor-induced lung cancer cell migration
http://www.unboundmedicine.com/medl…astasis_in_vivo

Smoking Cannabis Does Not Cause Cancer Of Lung or Upper Airways
http://ccrmg.org/journal/05aut/nocancer.html

No association between lung cancer and cannabis smoking in large study
http://www.cannabis-med.org/english…el.php?id=219#2

Marijuana Smoking Found Non-Carcinogenic
http://www.medpagetoday.com/Hematol…gCancer/tb/3393

CLAIM #4: MARIJUANA CAUSES LUNG DISEASE
http://www.erowid.org/plants/cannab…bis_myth4.shtml

Cancer- melanoma
Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases.
http://www.cannabis-med.org/studies…ow.php?s_id=180

Intractable nausea and vomiting due to gastrointestinal mucosal metastases
http://www.cannabis-med.org/studies…how.php?s_id=35

Cancer – oral
Smoking of cannabis does not increase risk for oral cancer
http://www.cannabis-med.org/english…el.php?id=175#1

Marijuana use and Risk of Oral Squamous Cell Carcinoma
http://66.218.69.11/search/cache?ei…&icp=1&.intl=us

Cancer-pancreatic
Cannabinoids Induce Apoptosis of Pancreatic Tumor Cells
http://cancerres.aacrjournals.org/c…ract/66/13/6748

Cancer – prostate
Inhibition of Human Breast and Prostate Cancer Cell Proliferation
http://endo.endojournals.org/cgi/co…tract/141/1/118

Cannabinoid Receptor as a Novel Target for the Treatment of Prostate Cancer
http://cancerres.aacrjournals.org/c…t/65/5/1635.pdf

——- Page 3

Cancer – Risk Cannabis vs Tobacco
Cannabis Smoke and Cancer: Assessing the Risk
http://www.norml.org/index.cfm?Group_ID=6891

Cannabis and tobacco smoke are not equally carcinogenic
http://www.pubmedcentral.nih.gov/ar…i?artid=1277837

Smoking Marijuana Does Not Cause Lung Cancer
http://www.mapinc.org/drugnews/v05/n1065/a03.html

Blunt Smokers Link Dependence Potential To Nicotine
http://www.medicalnewstoday.com/articles/52838.php

Premiere British Medical Journal Pronounces Marijuana Safer Than Alcohol, Tobacco
http://cannabislink.ca/medical/safer.html

Why Doesn’t Smoking Marijuana Cause Cancer?
http://www.healthcentral.com/drdean/408/14275.html

Marijuana Smoking Found Non-Carcinogenic
http://www.medpagetoday.com/Hematol…gCancer/tb/3393

Cancer – Skin
Inhibition of skin tumor growth
http://www.jci.org/cgi/content/full…y=MpUgjDbqHybAU

Cannabis Reduces Skin Cancer
http://www.onlinepot.org/medical/skincancerreport.htm

Cancer – Testicular
The antiemetic efficacy of nabilone
http://www.cannabis-med.org/studies…ow.php?s_id=127

Chemotherapy for Testicular Cancer
http://www.rxmarihuana.com/shared_c…icularchemo.htm

Cancer –various/ unnamed
Derivatives of cannabis for anti-cancer treatment
http://www.eurekalert.org/pub_relea…uo-do060605.php

Cancer Killer
http://www.november.org/stayinfo/br…ncerKiller.html

Anandamide Induces Apoptosis
http://www.jbc.org/cgi/content/abstract/275/41/31938

Nabilone improves pain and symptom management
http://www.cannabis-med.org/studies…ow.php?s_id=177

The effects of smoked cannabis in painful peripheral neuropathy
http://www.cannabis-med.org/studies…how.php?s_id=96

Delta-9-tetrahydrocannabinol for appetite stimulation
http://www.cannabis-med.org/studies…how.php?s_id=52

Dronabinol and prochlorperazine in combination
http://www.cannabis-med.org/studies…how.php?s_id=28

Dronabinol enhancement of appetite in cancer patients.
http://www.cannabis-med.org/studies…ow.php?s_id=149

Efficacy of tetrahydrocannabinol
http://www.cannabis-med.org/studies…how.php?s_id=31

Inhalation marijuana as an antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=155

Nabilone versus domperidone
http://www.cannabis-med.org/studies…ow.php?s_id=129

Inhalation marijuana as an antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=155

Nabilone vs. placebo in chemotherapy-induced nausea and vomiting
http://www.cannabis-med.org/studies…ow.php?s_id=156

The antiemetic activity of tetrahydrocanabinol versus metoclopramide
http://www.cannabis-med.org/studies…how.php?s_id=24

Delta-9-tetrahydrocannabinol as an antiemetic for patients receiving cancer chemotherapy
http://www.cannabis-med.org/studies…show.php?s_id=5

Delta-9-tetrahydrocannabinol as an antiemetic in cancer patients receiving high-dose methotrexate
http://www.cannabis-med.org/studies…how.php?s_id=23

Delta-9-tetrahydrocannabinol (THC) as an antiemetic in patients treated with cancer chemotherapy
http://www.cannabis-med.org/studies…how.php?s_id=27

Amelioration of cancer chemotherapy-induced nausea and vomiting by delta-9-THC
http://www.cannabis-med.org/studies…ow.php?s_id=107

Superiority of nabilone over prochlorperazine as an antiemetic
http://www.cannabis-med.org/studies…ow.php?s_id=126

Analgesic effect of delta-9-tetrahydrocannabinol.
http://www.cannabis-med.org/studies…how.php?s_id=16

The analgesic properties of delta-9-tetrahydrocannabinol and codeine.
http://www.cannabis-med.org/studies…how.php?s_id=17

Comparison of orally administered cannabis extract and delta-9-THC
http://www.unboundmedicine.com/medl…xia_Study_Group

Cannabis May Help Combat Cancer-causing Herpes Viruses
http://www.sciencedaily.com/release…40923092627.htm

Marijuana Smoking Found Non-Carcinogenic
http://www.medpagetoday.com/Hematol…gCancer/tb/3393

Cannabidiol
Cannabidiol, Antioxidants, and Diuretics in Reversing Binge Ethanol-Induced Neurotoxicity
http://jpet.aspetjournals.org/cgi/c…ourcetype=HWCIT

Cannabinol delays symptom onset
http://www.ncbi.nlm.nih.gov/sites/e…t_uids=16183560

Cannabidiol is an oral anti-arthritic therapeutic in murine collagen-induced arthritis
http://www.pnas.org/cgi/content/full/97/17/9561

Cannabidiol inhibits tumour growth in leukaemia and breast cancer
http://www.cannabis-med.org/english…el.php?id=220#2

Anti-tumor effects of cannabidiol
http://www.hempworld.com/HempPharm/…milanstudy.html

Cannabidiol triggers caspase activation and oxidative stress in human glioma cells.
http://www.ihop-net.org/UniPub/iHOP…l?pmid=16909207

Cannabidiol-Induced Apoptosis in Human Leukemia Cells
http://molpharm.aspetjournals.org/c…stract/70/3/897

Cannabidiol inhibits tumour growth in leukaemia and breast cancer
http://www.cannabis-med.org/english…el.php?id=220#2

Cannabidiol lowers incidence of diabetes in non-obese diabetic mice
http://www.ingentaconnect.com/conte…sn7o5efqr.alice

Neuroprotective and Blood-Retinal Barrier-Preserving Effects of Cannabidiol
http://ajp.amjpathol.org/cgi/content/full/168/1/235

Evaluation of cannabidiol in dystonic movement disorders
http://www.cannabis-med.org/studies…how.php?s_id=14

Cannabidiol in dystonic movement disorders.
http://www.cannabis-med.org/studies…ow.php?s_id=139

Beneficial and adverse effects of cannabidiol in a Parkinson patient
http://www.cannabis-med.org/studies…ow.php?s_id=142

Treatment of Meige’s syndrome with cannabidiol.
http://www.cannabis-med.org/studies…ow.php?s_id=114

CANNABIDIOL TO HEALTHY VOLUNTEERS AND EPILEPTIC PATIENTS
http://web.acsalaska.net/~warmgun/es201.html

Chronic administration of cannabidiol to healthy volunteers and epileptic patients.
http://www.cannabis-med.org/studies…how.php?s_id=42

Neuroprotective effect of (-)Delta9-tetrahydrocannabinol and cannabidiol
http://www.unboundmedicine.com/medl…f_peroxynitrite

EFFECTS OF CANNABIDIOL IN HUNTINGTON’S DISEASE
http://www.druglibrary.org/schaffer…al/hunting1.htm

The therapeutic rationale for combining tetrahydrocannabinol and cannabidiol.
http://www.medscape.com/medline/abstract/16209908

Cannabidiol has a cerebroprotective action
http://www.unboundmedicine.com/medl…iting_mechanism

Cannabidiol as an antipsychotic
http://www.cannabis-med.org/studies…ow.php?s_id=171

Cannabidiol, a constituent of Cannabis sativa, modulates sleep in rats.
http://www.medscape.com/medline/abs…844117?prt=true

Who’s Afraid of Cannabidiol?
http://www.counterpunch.org/gardner07142007.html

Chemical composition
Cannabis: A source of useful pharma compounds
http://www.medpot.net/forums/index.php?showtopic=18608

Pharmacokinetics and cannabinoid action using oral cannabis extract
http://www.pharma-lexicon.com/medic…hp?newsid=29638

Pharmacokinetics of cannabinoids
http://66.218.69.11/search/cache?ei…&icp=1&.intl=us

The chemistry and biological activity of cannabis
http://www.unodc.org/unodc/en/bulle….html?print=yes

Differential effects of medical marijuana based on strain and route of administration
http://www.medicalmarijuanaprocon.o…trainsstudy.pdf

What is THC?
http://www.medicalmarijuanaprocon.o…1.0373456855945

Cannabis / Marijuana ( ? 9 -Tetrahydrocannabinol, THC)
http://www.nhtsa.dot.gov/people/inj…gs/cannabis.htm

———- Page 4

Chemotherapy
Efficacy of dronabinol alone and in combination
http://www.cannabis-med.org/studies…ow.php?s_id=191

Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases
http://www.cannabis-med.org/studies…ow.php?s_id=180

Intractable nausea and vomiting
http://www.cannabis-med.org/studies…how.php?s_id=35

An efficient new cannabinoid antiemetic in pediatric oncology
http://www.cannabis-med.org/studies…show.php?s_id=7

Dronabinol and prochlorperazine in combination
http://www.cannabis-med.org/studies…how.php?s_id=28

Marijuana as antiemetic medicine
http://www.cannabis-med.org/studies…ow.php?s_id=134

Efficacy of tetrahydrocannabinol in patients refractory to standard anti-emetic therapy
http://www.cannabis-med.org/studies…how.php?s_id=31

Inhalation marijuana as an antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=155

Nabilone versus prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=120

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Antiemetic efficacy of nabilone and alizapride
http://www.cannabis-med.org/studies…ow.php?s_id=127

Nabilone versus domperidone
http://www.cannabis-med.org/studies…ow.php?s_id=129

THC or Compazine for the cancer chemotherapy patient
http://www.cannabis-med.org/studies…how.php?s_id=34

Comparison of nabilone and prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=128

Nabilone vs. prochlorperazine for refractory emesis
http://www.cannabis-med.org/studies…ow.php?s_id=146

Nabilone vs. placebo
http://www.cannabis-med.org/studies…ow.php?s_id=156

Tetrahydroannabinol (THC) vs prochlorperazine as chemotherapy antiemetics.
http://www.cannabis-med.org/studies…how.php?s_id=30

Comparative trial of the antiemetic effects of THC and haloperidol
http://www.cannabis-med.org/studies…how.php?s_id=64

Comparison of delta-9-tetrahydrocannabinol and prochlorperazine
http://www.cannabis-med.org/studies…show.php?s_id=3

Delta 9-tetrahydrocannabinol in cancer chemotherapy.
http://www.cannabis-med.org/studies…how.php?s_id=88

Antiemetic effect of tetrahydrocannabinol
http://www.cannabis-med.org/studies…show.php?s_id=6

Tetrahydrocanabinol versus metoclopramide and thiethylperazine
http://www.cannabis-med.org/studies…how.php?s_id=24

Effects of nabilone and prochlorperazine on chemotherapy-induced emesis
http://www.cannabis-med.org/studies…ow.php?s_id=131

Delta-9-tetrahydrocannabinol as an antiemetic
http://www.cannabis-med.org/studies…show.php?s_id=5

Delta-9-tetrahydrocannabinol as an antiemetic in cancer patients receiving high-dose methotrexate
http://www.cannabis-med.org/studies…how.php?s_id=23

THC as an antiemetic in patients treated with cancer chemotherapy
http://www.cannabis-med.org/studies…how.php?s_id=27

Amelioration of cancer chemotherapy-induced nausea and vomiting by delta-9-THC
http://www.cannabis-med.org/studies…ow.php?s_id=107

Superiority of nabilone over prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=126

Antiemetic effect of delta-9-tetrahydrocannabinol
http://www.cannabis-med.org/studies…show.php?s_id=4

Children
Experiences with THC-treatment in children and adolescents
http://www.cannabis-med.org/studies…how.php?s_id=80

An efficient new cannabinoid antiemetic in pediatric oncology.
http://www.cannabis-med.org/studies…show.php?s_id=7

Nabilone versus prochlorperazine for control of cancer chemotherapy-induced emesis in children
http://www.cannabis-med.org/studies…ow.php?s_id=120

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Marijuana and ADD Therapeutic uses of Medical Marijuana in the treatment of ADD
http://www.onlinepot.org/medical/add&mmj.htm

Oily fish makes ‘babies brainier’
http://news.bbc.co.uk/2/hi/health/4631006.stm

Cannabis is a First-Line Treatment for Childhood Mental Disorders
http://www.counterpunch.org/mikuriya07082006.html

Ganja use among Jamaican women.
http://www.rism.org/isg/dlp/ganja/a…anjaBabyes.html

Dreher’s Jamaican Pregnancy Study
http://www.november.org/stayinfo/br…reherStudy.html

Cannabis Relieves Morning Sickness
http://ccrmg.org/journal/06spr/dreher.html#morning

Moderate cannabis use not harmful to the brain of adolescents, M R I study finds
http://www.cannabis-med.org/english…el.php?id=218#3

No brain structural change associated with adolescent cannabis use
http://www.pubmedcentral.nih.gov/bo…l&artid=1524733

No ‘Smoking’ Gun: Research Indicates Teen Marijuana Use Does Not Predict Drug, Alcohol Abuse
http://www.sciencedaily.com/release…61204123422.htm

Pot May Not Shrink Teens’ Brains After All
http://www.medpagetoday.com/Neurolo…urology/tb/3242

Chronic Cystitis
Cannabinoid rotation in a young woman with chronic cystitis
http://www.cannabis-med.org/studies…ow.php?s_id=115

CPOD
THC effective in appetite and weight loss in severe lung disease (COPD)
http://www.cannabis-med.org/english…el.php?id=191#2

Heavy Long-Term Marijuana Use Does Not Impair Lung Function
http://www.erowid.org/plants/cannab…is_media7.shtml

Diabetes
Cannabinoid Reduces Incidence Of Diabetes
http://www.norml.org/index.cfm?Group_ID=6909

Marijuana Compound May Help Stop Diabetic Retinopathy
http://www.sciencedaily.com/release…60227184647.htm

Cannabidiol lowers incidence of diabetes in non-obese diabetic mice
http://www.ingentaconnect.com/conte…sn7o5efqr.alice

Anticoagulant Effects of a Cannabis Extract in an Obese Rat Model
http://www.level1diet.com/research/id/14687

Neuroprotective and Blood-Retinal Barrier-Preserving Effects of Cannabidiol
http://ajp.amjpathol.org/cgi/content/full/168/1/235

The Cannabinergic System as a Target for Anti-inflammatory Therapies
http://www.ingentaconnect.com/conte…000013/art00008

Effect of tetrahydrocurcumin on blood glucose, plasma insulin and hepatic key enzymes
http://www.unboundmedicine.com/medl…d_diabetic_rats

Cannabidiol reduces the development of diabetes in an animal study
http://www.cannabis-med.org/english…el.php?id=219#3

Depression
Cannabinoids promote hippocampus neurogenesis and produce anxiolytic- and antidepressant
http://www.jci.org/cgi/content/full/115/11/3104

Antidepressant-like activity by blockade of anandamide hydrolysis
http://www.pubmedcentral.nih.gov/ar…bmedid=16352709

Decreased depression in marijuana users.
http://www.medscape.com/medline/abstract/15964704

Antidepressant-like activity
http://www.pubmedcentral.nih.gov/ar…bmedid=16352709

Dronabinol and marijuana in HIV-positive marijuana smokers: caloric intake, mood, and sleep.
http://www.cannabis-med.org/studies…ow.php?s_id=190

Nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Cannabis and Depression
http://www.pacifier.com/~alive/cmu/…nd_cannabis.htm

Association between cannabis use and depression may not be causal, study says
http://www.cannabis-med.org/english…el.php?id=177#4

Marijuana use and depression among adults: Testing for causal associations.
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Do patients use marijuana as an antidepressant?
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Dermatitis
Efficacy of dietary hempseed oil in patients with atopic dermatitis.
http://www.medscape.com/medline/abs…ryText=hempseed

Dronabinol
Dronabinol in the treatment of agitation in patients with Alzheimer’s disease with anorexia
http://www.cannabis-med.org/studies…how.php?s_id=61

Dronabinol in the treatment of refractory agitation in Alzheimer’s disease
http://www.cannabis-med.org/studies…how.php?s_id=92

Effects of dronabinol on anorexia and disturbed behavior in patients with Alzheimer’s disease.
http://www.cannabis-med.org/studies…how.php?s_id=59

Dronabinol an effective appetite stimulant?
http://www.cannabis-med.org/studies…ow.php?s_id=188

Safety and efficacy of dronabinol in the treatment of agitation in patients with Alzheimer’s disease
http://www.cannabis-med.org/studies…how.php?s_id=61

Effect of dronabinol on nutritional status in HIV infection.
http://www.cannabis-med.org/studies…ow.php?s_id=150

Dronabinol stimulates appetite and causes weight gain in HIV patients.
http://www.cannabis-med.org/studies…how.php?s_id=20

Dronabinol effects on weight in patients with HIV infection.
http://www.cannabis-med.org/studies…how.php?s_id=45

Recent clinical experience with dronabinol.
http://www.cannabis-med.org/studies…how.php?s_id=90

Dronabinol enhancement of appetite in cancer patients.
http://www.cannabis-med.org/studies…ow.php?s_id=149

Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases.
http://www.cannabis-med.org/studies…ow.php?s_id=180

Dronabinol and prochlorperazine in combination
http://www.cannabis-med.org/studies…how.php?s_id=28

Dronabinol enhancement of appetite in cancer patients.
http://www.cannabis-med.org/studies…ow.php?s_id=149

Efficacy of dronabinol alone and in combination
http://www.cannabis-med.org/studies…ow.php?s_id=191

Dronabinol and marijuana in HIV-positive marijuana smokers: caloric intake, mood, and sleep.
http://www.cannabis-med.org/studies…ow.php?s_id=190

Dronabinol and retinal hemodynamics in humans.
http://www.cannabis-med.org/studies…ow.php?s_id=202

Dronabinol reduces signs and symptoms of idiopathic intracranial hypertension
http://www.cannabis-med.org/studies…ow.php?s_id=181

Nausea relieved by tetrahydrocannabinol (dronabinol).
http://www.cannabis-med.org/studies…how.php?s_id=35

Dronabinol in patients with intractable pruritus secondary to cholestatic liver disease.
http://www.cannabis-med.org/studies…ow.php?s_id=116

Treatment of spasticity in spinal cord injury with dronabinol
http://www.cannabis-med.org/studies…ow.php?s_id=112

Cannabinoid Activator Mellows Out Colon
http://www.medpagetoday.com/MeetingCoverage/ACG/tb/4410

Drug testing
Hemp oil causes positive urine tests for THC
http://www.druglibrary.org/crl/drug…0JAnToxicol.pdf

Dystonia
Cannabis sativa and dystonia secondary to Wilson’s disease.
http://www.medscape.com/medline/abstract/15390041

Experiences with THC-treatment in children and adolescents
http://www.cannabis-med.org/studies…how.php?s_id=80

Evaluation of cannabidiol in dystonic movement disorders
http://www.cannabis-med.org/studies…how.php?s_id=14

Cannabidiol in dystonic movement disorders.
http://www.cannabis-med.org/studies…ow.php?s_id=139

Beneficial and adverse effects of cannabidiol in a Parkinson patient
http://www.cannabis-med.org/studies…ow.php?s_id=142

Treatment of Meige’s syndrome with cannabidiol.
http://www.cannabis-med.org/studies…ow.php?s_id=114

———- Page 5

Endocannabinoid Deficiency
Clinical Endocannabinoid Deficiency
http://www.freedomtoexhale.com/clinical.pdf

The endocannabinoid system is dysregulated in multiple sclerosis
http://brain.oxfordjournals.org/cgi…stract/awm160v1

Cannabinoids inhibit neurodegeneration in models of multiple sclerosis
http://brain.oxfordjournals.org/cgi…ull/126/10/2191

Epilepsy
Epilepsy patients are smoking pot
http://www.safeaccessnow.org/article.php?id=1638

CANNABIDIOL TO HEALTHY VOLUNTEERS AND EPILEPTIC PATIENTS
http://web.acsalaska.net/~warmgun/es201.html

Experiences with THC-treatment in children and adolescents
http://www.cannabis-med.org/studies…how.php?s_id=80

Chronic administration of cannabidiol to healthy volunteers and epileptic patients.
http://www.cannabis-med.org/studies…how.php?s_id=42

Anticonvulsant nature of marihuana smoking.
http://www.cannabis-med.org/studies…how.php?s_id=39

Cannabis may help epileptics
http://www.medicalnewstoday.com/articles/4423.php

Hypnotic and Antiepileptic Effects of Cannabidiol
http://www.thecompassionclub.org/me…rue&pageNumber=

Marijuana: an effective antiepileptic treatment in partial epilepsy?
http://www.cannabis-med.org/studies…ow.php?s_id=157

Familial Mediterranean Fever
Pain relief with oral cannabinoids in familial Mediterranean fever.
http://www.cannabis-med.org/studies…how.php?s_id=18

Fertility
Synthetic Cannabinoid May Aid Fertility In Smokers
http://www.medicalnewstoday.com/articles/58063.php

Medical marijuana uses – 700 medical marijuana clinical studies and papers

Fever
A Novel Role of Cannabinoids
http://ccicnewsletter.com/index.php…nfectious_Disea

A Cooling Effect From Cannabis?
http://ccrmg.org/journal/05aut/coolcannabis.html

Fibromyalgia
Delta-9-THC based monotherapy in fibromyalgia patients
http://www.medscape.com/medline/abstract/16834825

Clinical Endocannabinoid Deficiency
http://www.freedomtoexhale.com/clinical.pdf

Cannabis Sativa (Marijuana) for Fibromyalgia
http://www.fibromyalgia-reviews.com/Drg_Marijuana.cfm

THC Reduces Pain in Fibromyalgia Patients
http://www.illinoisnorml.org/content/view/63/35/

Gateway Theory
The Myth of Marijuana’s Gateway Effect
http://www.druglibrary.org/schaffer/library/mjgate.htm

Endogenous cannabinoids are not involved in cocaine reinforcement
http://www.sciencedirect.com/scienc…a4e861a90579fac

No ‘Smoking’ Gun: Research Indicates Teen Marijuana Use Does Not Predict Drug, Alcohol Abuse
http://www.sciencedaily.com/release…61204123422.htm

CLAIM #13:MARIJUANA IS A “GATEWAY” TO THE USE OF OTHER DRUGS
http://www.erowid.org/plants/cannab…is_myth13.shtml

Glaucoma
Marijuana Smoking vs Cannabinoids for Glaucoma Therapy
http://archopht.ama-assn.org/cgi/co…act/116/11/1433

Dronabinol and retinal hemodynamics in humans.
http://www.cannabis-med.org/studies…ow.php?s_id=202

Effect of Sublingual Application of Cannabinoids on Intraocular Pressure
http://www.cannabis-med.org/studies…ow.php?s_id=201

Delta 9-tetrahydrocannabinol in cancer chemotherapy. Ophthalmologic implications.
http://www.cannabis-med.org/studies…how.php?s_id=88

Effect of marihuana on intraocular and blood pressure in glaucoma.
http://www.cannabis-med.org/studies…how.php?s_id=87

Effect of delta-9-tetrahydrocannabinol on intraocular pressure in humans.
http://www.cannabis-med.org/studies…how.php?s_id=40

Marihuana smoking and intraocular pressure.
http://www.cannabis-med.org/studies…how.php?s_id=47

Neuroprotective and Intraocular Pressure-Lowering Effects of (-)Delta-Tetrahydrocannabinol
http://www.unboundmedicine.com/medl…del_of_Glaucoma

Neuroprotective effect of (-)Delta9-tetrahydrocannabinol and cannabidiol
http://www.unboundmedicine.com/medl…f_peroxynitrite

Effects of tetrahydrocannabinol on arterial and intraocular hypertension.
http://www.medscape.com/medline/abstract/468444

Gynocology and obstetrics
Cannabis Treatments in Obstetrics and Gynecology: A Historical Review
http://www.freedomtoexhale.com/russo-ob.pdf

Heart Disease/ Cardiovascular
Marijuana Chemical Fights Hardened Arteries
http://www.webmd.com/heart-disease/…rdened-arteries

The endogenous cardiac cannabinoid system: a new protective mechanism
http://www.cannabinoid.com/boards/thd3x10073.shtml

Cardiovascular pharmacology of cannabinoids.
http://www.biowizard.com/story.php?pmid=16596789

Delta-9-tetrahydrocannabinol protects cardiac cells from hypoxia
http://www.ingentaconnect.com/conte…020001/00002346

Does Cannabis Hold the Key to Treating Cardiometabolic Disease?
http://www.medscape.com/viewarticle/525040_print

Cannabinoid Offers Cardioprotection
http://www.norml.org/index.cfm?Grou…tm_format=print

Heavy Cannabis Use Not Independently Associated With Cardiovascular Risks
http://www.norml.org/index.cfm?Group_ID=6972

Marijuana use, diet, body mass index, and cardiovascular risk factors
http://www.medscape.com/medline/abstract/16893701

Cannabinoids and cardiovascular disease
http://www.unboundmedicine.com/medl…ical_treatments

Cannabinoids as therapeutic agents in cardiovascular disease
http://www.unboundmedicine.com/medl…s_and_illusions

The in vitro and in vivo cardiovascular effects of {Delta}9-tetrahydrocannabinol
http://www.unboundmedicine.com/medl…_oxide_synthase

Cannabinoids prevented the development of heart failure in animal study
http://www.cannabis-med.org/english…el.php?id=145#2

Cannabis use not associated with risk factors for diseases of heart and circulation
http://www.cannabis-med.org/english…el.php?id=225#2

THC protects heart cells in the case of lowered oxygen supply
http://www.cannabis-med.org/english…el.php?id=212#1

Medical marijuana: study shows that THC slows atherosclerosis
http://thenexthurrah.typepad.com/th…al_marijua.html

Cardiovascular Effects of Cannabis
http://www.idmu.co.uk/canncardio.htm

Changes in middle cerebral artery velocity after marijuana
http://www.ncbi.nlm.nih.gov/sites/e…0&dopt=Abstract[/]

—– Page 6

Hepatitis
Moderate Cannabis Use Associated with Improved Treatment Response
http://www.hivandhepatitis.com/hep_…6/091506_a.html

Cannabis use improves retention and virological outcomes in patients treated for hepatitis C
http://www.natap.org/2006/HCV/091506_02.htm

Hepatitis C – The Silent Killer Can Medical Cannabis Help?
http://www.pacifier.com/~alive/cmu/hepatitis_c.htm

Herpes
Cannabis May Help Combat Cancer-causing Herpes Viruses
http://www.sciencedaily.com/release…40923092627.htm

THC inhibits lytic replication of gamma oncogenic herpes viruses in vitro
http://www.pubmedcentral.nih.gov/bo…ml&artid=521080

Suppressive effect of delta-9-tetrahydrocannabinol on herpes simplex virus infectivity in vitro
http://www.ebmonline.org/cgi/content/abstract/196/4/401

Inhibition of cell-associated herpes simplex virus
http://www.ebmonline.org/cgi/content/abstract/185/1/41

The Effect of {Delta}-9-Tetrahydrocannabinol on Herpes Simplex Virus Replication
http://vir.sgmjournals.org/cgi/cont…stract/49/2/427

Hiccups
Marijuana cures hiccups
http://www.yourhealthbase.com/database/a77k.htm

Marijuana For Intractable Hiccups
http://cannabislink.ca/medical/hiccups.html

HIV / AIDS
Marijuana Use Does Not Accelerate HIV Infection
http://paktribune.com/news/print.php?id=139255

THC improves appetite and reverses weight loss in AIDS patients
http://www.cannabis-med.org/studies…ow.php?s_id=189

Dronabinol and marijuana in HIV-positive marijuana smokers: caloric intake, mood, and sleep.
http://www.cannabis-med.org/studies…ow.php?s_id=190

Cannabis in painful HIV-associated sensory neuropathy
http://www.cannabis-med.org/studies…ow.php?s_id=199

Smoked cannabis therapy for HIV-related painful peripheral neuropathy
http://www.cannabis-med.org/studies…ow.php?s_id=172

Short-term effects of cannabinoids in patients with HIV-1 infection
http://www.cannabis-med.org/studies…how.php?s_id=62

Dronabinol as a treatment for anorexia associated with weight loss in patients with AIDS.
http://www.cannabis-med.org/studies…how.php?s_id=21

Effect of dronabinol on nutritional status in HIV infection.
http://www.cannabis-med.org/studies…ow.php?s_id=150

Dronabinol stimulates appetite and causes weight gain in HIV patients.
http://www.cannabis-med.org/studies…how.php?s_id=20

Dronabinol effects on weight in patients with HIV infection.
http://www.cannabis-med.org/studies…how.php?s_id=45

Recent clinical experience with dronabinol.
http://www.cannabis-med.org/studies…how.php?s_id=90

Marijuana as therapy for people living with HIV/AIDS: Social and health aspects
http://www.unboundmedicine.com/medl…_health_aspects

Marijuana and AIDS: A Four-Year Study
http://ccrmg.org/journal/05spr/aids.html

Historical studies
The La Guardia Committee Report
http://www.druglibrary.org/schaffer…lag/lagmenu.htm

Physical, Mental, and Moral Effects of Marijuana: The Indian Hemp Drugs Commission Report
http://www.druglibrary.org/schaffer/Library/effects.htm

MARIAJUANA SMOKING IN PANAMA
http://www.druglibrary.org/schaffer…ama/panama1.htm

The British Pharmaceutical Codex – 1934
http://www.druglibrary.org/schaffer…ical/brit34.htm

ON THE PREPARATIONS OF THE INDIAN HEMP, OR GUNJAH
http://www.druglibrary.org/schaffer…1850/gunjah.htm

DISPENSATORY OF THE UNITED STATES OF AMERICA Fifth Edition (1843)
http://www.druglibrary.org/schaffer…ry/dispensa.htm

New Remedies:Pharmaceutically and Therapeutically Considered Fourth Edition (1843)
http://www.druglibrary.org/schaffer…ry/dunglisn.htm

On the Haschisch or Cannabis Indica
http://www.druglibrary.org/schaffer…ry/bellhash.htm

ON INDICATIONS OF THE HACHISH-VICE IN THE OLD TESTAMENT
http://www.druglibrary.org/schaffer…tory/hashot.htm

The Physiological Activity of Cannabis Sativa
http://www.druglibrary.org/schaffer…istory/japa.htm

CANNABIS, U.S.P. (American Cannabis):
http://www.druglibrary.org/schaffer…ry/vbchmed1.htm

Hormones
Effects of chronic marijuana use on testosterone, luteinizing hormone, follicle stimulating …
http://www.anesth.uiowa.edu/readabs…sp?PMID=1935564

Marijuana: interaction with the estrogen receptor
http://jpet.aspetjournals.org/cgi/c…tract/224/2/404

Huntington’s Disease
EFFECTS OF CANNABIDIOL IN HUNTINGTON’S DISEASE
http://www.druglibrary.org/schaffer…al/hunting1.htm

Nabilone Could Treat Chorea and Irritability in Huntington’s Disease
http://neuro.psychiatryonline.org/c…/18/4/553?rss=1

Hysterectomy
Effect of nabilone on nausea and vomiting after total abdominal hysterectomy
http://www.cannabis-med.org/studies…ow.php?s_id=137

Idiopathic Intracranial Hypertension
Dronabinol reduces signs and symptoms of idiopathic intracranial hypertension
http://www.cannabis-med.org/studies…ow.php?s_id=181

IQ
Findings of a longitudinal study of effects on IQ
http://www.cmaj.ca/cgi/content/full/166/7/887

Heavy cannabis use without long-term effect on global intelligence
http://www.cannabis-med.org/english…el.php?id=115#2

Marijuana does not dent IQ permanently
http://www.newscientist.com/article…ermanently.html

Marinol/Synthetics/ cannabinoid mixtures
CANNABIS AND MARINOL IN THE TREATMENT OF MIGRAINE HEADACHE
http://www.druglibrary.org/schaffer/hemp/migrn2.htm

Marinol vs Natural Cannabis
http://www.norml.org/pdf_files/NORM…al_Cannabis.pdf

The therapeutic rationale for combining tetrahydrocannabinol and cannabidiol.
http://www.medscape.com/medline/abstract/16209908

Unheated Cannabis sativa extracts and its major compound THC-acid
http://www.medscape.com/medline/abs…504929?prt=true

Side effects of pharmaceuticals not elicited by comparable herbal medicines.
http://www.medscape.com/medline/abstract/10394675

Sativex in the treatment of pain caused by rheumatoid arthritis
http://rheumatology.oxfordjournals….bstract/45/1/50

Is dronabinol an effective appetite stimulant?
http://www.cannabis-med.org/studies…ow.php?s_id=188

Sativex in patients suffering from multiple sclerosis associated detrusor overactivity
http://www.cannabis-med.org/studies…ow.php?s_id=168

Sativex® in patients with symptoms of spasticity due to multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=169

Nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases
http://www.cannabis-med.org/studies…ow.php?s_id=180

Synthetic cannabinomimetic nabilone on patients with chronic pain
http://www.cannabis-med.org/studies…ow.php?s_id=197

Nabilone significantly reduces spasticity-related pain
http://www.cannabis-med.org/studies…ow.php?s_id=200

Sativex produced significant improvements in a subjective measure of spasticity
http://www.cannabis-med.org/studies…ow.php?s_id=170

Analgesic effect of the synthetic cannabinoid CT-3 on chronic neuropathic pain
http://www.cannabis-med.org/studies…how.php?s_id=85

Cannabinoid rotation in a young woman with chronic cystitis
http://www.cannabis-med.org/studies…ow.php?s_id=115

Dronabinol in patients with intractable pruritus
http://www.cannabis-med.org/studies…ow.php?s_id=116

Cannabinoids reduce levodopa-induced dyskinesia in Parkinson’s disease:
http://www.cannabis-med.org/studies…how.php?s_id=54

Nabilone on L-DOPA induced dyskinesia in patients with idiopathic Parkinson’s disease
http://www.cannabis-med.org/studies…ow.php?s_id=153

Nabilone in the treatment of multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=11

Big Pharma’s Strange Holy Grail: Cannabis Without Euphoria?
http://www.counterpunch.org/gardner07082006.html

Sativex showed positive effects in 65 per cent of patients with chronic diseases
http://www.cannabis-med.org/english…el.php?id=230#4

Meige’s Syndrome
Treatment of Meige’s syndrome with cannabidiol.
http://www.cannabis-med.org/studies…ow.php?s_id=114

Migraine/ Headache
CANNABIS AND MARINOL IN THE TREATMENT OF MIGRAINE HEADACHE
http://www.druglibrary.org/schaffer/hemp/migrn2.htm

Dronabinol reduces signs and symptoms of idiopathic intracranial hypertension
http://www.cannabis-med.org/studies…ow.php?s_id=181

Cannabis and Migraine
http://www.pacifier.com/~alive/cmu/…nd_migraine.htm

Clinical Endocannabinoid Deficiency
http://www.freedomtoexhale.com/clinical.pdf

Hemp for Headache
http://www.freedomtoexhale.com/hh.pdf

Chronic Migraine Headache
http://www.druglibrary.org/schaffer/hemp/migrn1.htm

Morning Sickness
Medical marijuana: a surprising solution to severe morning sickness http://www.findarticles.com/p/artic…124/ai_n6015580

Medicinal cannabis use among childbearing women
http://safeaccess.ca/research/cannabis_nausea2006.pdf

Mortality Rates
Marijuana use and mortality.
http://www.pubmedcentral.nih.gov/ar…i?artid=1380837

Marijuana Smoking Doesn’t Lead to Higher Death Rate
http://ccrmg.org/journal/03sum/kaiser.html

How deadly is marijuana?
http://www.medicalnewstoday.com/articles/4426.php

———– Page 7

MS
Sativex in patients with symptoms of spasticity due to multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=169

Marijuana derivatives may provide MS treatment
http://www.healthypages.net/news.asp?newsid=5381

Marijuana Helps MS Patients Alleviate Pain, Spasms
http://www.mult-sclerosis.org/news/…smsAndPain.html

Cannabis-based medicine in central pain in multiple sclerosis
http://www.neurology.org/cgi/conten…t/65/6/812?etoc

Cannabis-based medicine in spasticity caused by multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=192

Sativex in patients suffering from multiple sclerosis associated detrusor overactivity
http://www.cannabis-med.org/studies…ow.php?s_id=168

The effect of cannabis on urge incontinence in patients with multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=185

Nabilone significantly reduces spasticity-related pain
http://www.cannabis-med.org/studies…ow.php?s_id=200

Cannabinoids in multiple sclerosis (CAMS) study
http://www.cannabis-med.org/studies…ow.php?s_id=160

Sativex produced significant improvements in a subjective measure of spasticity
http://www.cannabis-med.org/studies…ow.php?s_id=170

Cannabis-based medicine in central pain in multiple sclerosis.
http://www.cannabis-med.org/studies…ow.php?s_id=175

Do cannabis-based medicinal extracts have general or specific effects
http://www.cannabis-med.org/studies…how.php?s_id=56

Efficacy, safety and tolerability of an oral cannabis extract in the treatment of spasticity
http://www.cannabis-med.org/studies…how.php?s_id=63

cannabis-based extracts for bladder dysfunction in advanced multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=81

Are oral cannabinoids safe and effective in refractory neuropathic pain?
http://www.cannabis-med.org/studies…ow.php?s_id=143

Dronabinol in the treatment of agitation in patients with Alzheimer’s disease with anorexia
http://www.cannabis-med.org/studies…how.php?s_id=61

Cannabis based medicinal extracts (CBME) in central neuropathic pain due to multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=82

Cannabinoids for treatment of spasticity and other symptoms related to multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=108

Cannabis based medicinal extract on refractory lower urinary tract dysfunction
http://www.cannabis-med.org/studies…ow.php?s_id=103

Analgesic effect of the cannabinoid analogue nabilone
http://www.cannabis-med.org/studies…ow.php?s_id=203

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

Nabilone in the treatment of multiple sclerosis
http://www.cannabis-med.org/studies…how.php?s_id=11

Effect of cannabinoids on spasticity and ataxia in multiple sclerosis.
http://www.cannabis-med.org/studies…show.php?s_id=2

Delta-9-THC in the treatment of spasticity associated with multiple sclerosis.
http://www.cannabis-med.org/studies…show.php?s_id=1

Tetrahydrocannabinol for tremor in multiple sclerosis.
http://www.cannabis-med.org/studies…show.php?s_id=9

Marihuana as a therapeutic agent for muscle spasm or spasticity
http://www.cannabis-med.org/studies…how.php?s_id=53

Cannabis-based medicine in spasticity caused by multiple sclerosis.
http://www.unboundmedicine.com/medl…tiple_sclerosis

Cannabis based treatments for neuropathic and multiple sclerosis-related pain.
http://www.unboundmedicine.com/medl…is_related_pain

The effect of cannabis on urge incontinence in patients with multiple sclerosis
http://www.unboundmedicine.com/medl…ial__CAMS_LUTS_

Can Cannabis Help Multiple Sclerosis? An International Debate Rages
http://www.pacifier.com/~alive/cmu/…bis_help_ms.htm

Cannabis’ Potential Exciting Researchers in Treatment of ALS, Parkinson’s Disease
http://66.218.69.11/search/cache?ei…&icp=1&.intl=us

The endocannabinoid system is dysregulated in multiple sclerosis
http://brain.oxfordjournals.org/cgi…stract/awm160v1

Cannabinoids inhibit neurodegeneration in models of multiple sclerosis
http://brain.oxfordjournals.org/cgi…ull/126/10/2191

Nabilone
The synthetic cannabinoid nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

Nabilone versus prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=120

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Antiemetic efficacy of nabilone and alizapride
http://www.cannabis-med.org/studies…ow.php?s_id=127

Nabilone versus domperidone
http://www.cannabis-med.org/studies…ow.php?s_id=129

Comparison of nabilone and prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=128

Nabilone vs. prochlorperazine for refractory emesis
http://www.cannabis-med.org/studies…ow.php?s_id=146

Nabilone vs. placebo
http://www.cannabis-med.org/studies…ow.php?s_id=156

Effects of nabilone and prochlorperazine on chemotherapy-induced emesis
http://www.cannabis-med.org/studies…ow.php?s_id=131

Superiority of nabilone over prochlorperazine
http://www.cannabis-med.org/studies…ow.php?s_id=126

Nabilone versus prochlorperazine for control of cancer chemotherapy-induced emesis in children
http://www.cannabis-med.org/studies…ow.php?s_id=120

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Effect of nabilone on nausea and vomiting after total abdominal hysterectomy
http://www.cannabis-med.org/studies…ow.php?s_id=137

Nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

Synthetic cannabinomimetic nabilone on patients with chronic pain
http://www.cannabis-med.org/studies…ow.php?s_id=197

Nabilone significantly reduces spasticity-related pain
http://www.cannabis-med.org/studies…ow.php?s_id=200

Nabilone on L-DOPA induced dyskinesia in patients with idiopathic Parkinson’s disease
http://www.cannabis-med.org/studies…ow.php?s_id=153

Nabilone in the treatment of multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=11

Nabilone significantly reduces spasticity-related pain
http://www.cannabis-med.org/studies…ow.php?s_id=200

Analgesic effect of the cannabinoid analogue nabilone
http://www.cannabis-med.org/studies…ow.php?s_id=203

Nabilone in the treatment of multiple sclerosis
http://www.cannabis-med.org/studies…how.php?s_id=11

Comparison of nabilone and metoclopramide in the control of radiation-induced nausea.
http://www.cannabis-med.org/studies…ow.php?s_id=130

Nabilone and metoclopramide in the treatment of nausea and vomiting
http://www.cannabis-med.org/studies…ow.php?s_id=121

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Comparison of the antiemetic efficacy of nabilone and alizapride
http://www.cannabis-med.org/studies…ow.php?s_id=127

Nabilone versus domperidone in the treatment of cytotoxic-induced emesis.
http://www.cannabis-med.org/studies…ow.php?s_id=129

Add-on treatment with the synthetic cannabinomimetic nabilone on patients with chronic pain -
http://www.cannabis-med.org/studies…ow.php?s_id=197

Comparison of bronchial effects of nabilone and terbutaline
http://www.cannabis-med.org/studies…how.php?s_id=43

Nabilone Could Treat Chorea and Irritability in Huntington’s Disease
http://neuro.psychiatryonline.org/c…/18/4/553?rss=1

Nausea
THC improves appetite and reverses weight loss in AIDS patients
http://www.cannabis-med.org/studies…ow.php?s_id=189

Efficacy of dronabinol alone and in combination with ondansetron versus ondansetron alone
http://www.cannabis-med.org/studies…ow.php?s_id=191

Dronabinol and marijuana in HIV-positive marijuana smokers: caloric intake, mood, and sleep.
http://www.cannabis-med.org/studies…ow.php?s_id=190

Nabilone improves pain and symptom management in cancer patients
http://www.cannabis-med.org/studies…ow.php?s_id=177

Dronabinol for supportive therapy in patients with malignant melanoma and liver metastases.
http://www.cannabis-med.org/studies…ow.php?s_id=180

Nausea relieved by tetrahydrocannabinol (dronabinol).
http://www.cannabis-med.org/studies…how.php?s_id=35

An efficient new cannabinoid antiemetic in pediatric oncology.
http://www.cannabis-med.org/studies…show.php?s_id=7

Effect of nabilone on nausea and vomiting after total abdominal hysterectomy.
http://www.cannabis-med.org/studies…ow.php?s_id=137

Marijuana as antiemetic medicine
http://www.cannabis-med.org/studies…ow.php?s_id=134

Efficacy of tetrahydrocannabinol in patients refractory to standard anti-emetic therapy
http://www.cannabis-med.org/studies…how.php?s_id=31

Inhalation marijuana as an antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=155

Nabilone versus prochlorperazine for control of cancer chemotherapy-induced emesis in children
http://www.cannabis-med.org/studies…ow.php?s_id=120

Comparison of nabilone and metoclopramide in the control of radiation-induced nausea.
http://www.cannabis-med.org/studies…ow.php?s_id=130

Nabilone and metoclopramide in the treatment of nausea and vomiting
http://www.cannabis-med.org/studies…ow.php?s_id=121

Nabilone: an alternative antiemetic for cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=123

Comparison of the antiemetic efficacy of nabilone and alizapride
http://www.cannabis-med.org/studies…ow.php?s_id=127

Nabilone versus domperidone in the treatment of cytotoxic-induced emesis.
http://www.cannabis-med.org/studies…ow.php?s_id=129

THC or Compazine for the cancer chemotherapy patient–the UCLA study
http://www.cannabis-med.org/studies…how.php?s_id=34

Comparison of nabilone and prochlorperazine for emesis induced by cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=128

Acute and subacute bronchial effects of oral cannabinoids.
http://www.cannabis-med.org/studies…how.php?s_id=44

Nabilone vs. prochlorperazine for refractory emesis induced by cancer chemotherapy.
http://www.cannabis-med.org/studies…ow.php?s_id=146

Nabilone vs. placebo in chemotherapy-induced nausea and vomiting.
http://www.cannabis-med.org/studies…ow.php?s_id=156

Dose vs response of tetrahydroannabinol (THC) vs prochlorperazine
http://www.cannabis-med.org/studies…how.php?s_id=30 delta 9-

Comparative trial of the antiemetic effects of THC and haloperidol
http://www.cannabis-med.org/studies…how.php?s_id=64

Comparison of delta-9-tetrahydrocannabinol and prochlorperazine.
http://www.cannabis-med.org/studies…show.php?s_id=3

Tetrahydrocannabinol in cancer chemotherapy. Ophthalmologic implications.
http://www.cannabis-med.org/studies…how.php?s_id=88

Antiemetic effect of tetrahydrocannabinol
http://www.cannabis-med.org/studies…show.php?s_id=6

The antiemetic activity of tetrahydrocanabinol versus metoclopramide and thiethylperazine
http://www.cannabis-med.org/studies…how.php?s_id=24

The antiemetic effects of nabilone and prochlorperazine on chemotherapy-induced emesis.
http://www.cannabis-med.org/studies…ow.php?s_id=131

Delta-9-tetrahydrocannabinol as an antiemetic for patients receiving cancer chemotherapy
http://www.cannabis-med.org/studies…show.php?s_id=5

Delta-9-tetrahydrocannabinol as an antiemetic in cancer patients receiving high-dose methotrexate
http://www.cannabis-med.org/studies…how.php?s_id=23

THC as an antiemetic in patients treated with cancer chemotherapy
http://www.cannabis-med.org/studies…how.php?s_id=27

Amelioration of cancer chemotherapy-induced nausea and vomiting by delta-9-THC.
http://www.cannabis-med.org/studies…ow.php?s_id=107

Superiority of nabilone over prochlorperazine as an antiemetic
http://www.cannabis-med.org/studies…ow.php?s_id=126

Antiemetic effect of delta-9-tetrahydrocannabinol in patients receiving cancer chemotherapy.
http://www.cannabis-med.org/studies…show.php?s_id=4

Receptor mechanism and antiemetic activity of structurally-diverse cannabinoids
http://www.unboundmedicine.com/medl…the_least_shrew

Neurons
Marijuana Promotes Neuron Growth
http://www.medpot.net/forums/index.php?showtopic=27460

Marijuana-Like Chemicals in the Brain Calm Neurons
http://www.medpot.net/forums/index.php?showtopic=9686

Marijuana May Spur New Brain Cells
http://www.treatingyourself.com/vbu…read.php?t=5921

Cannabinoids promote embryonic and adult hippocampus neurogenesis
http://www.jci.org/cgi/content/full/115/11/3104

Medical marijuana uses – 700 medical marijuana clinical studies and papers

————— Page 8

Neuropathic pain
Cannabinoids Among Most Promising Approaches to Treating Neuropathic Pain
http://www.redorbit.com/news/health…source=r_health

Cannabis-based medicine in central pain in multiple sclerosis
http://www.neurology.org/cgi/conten…t/65/6/812?etoc

Cannabis in painful HIV-associated sensory neuropathy
http://www.cannabis-med.org/studies…ow.php?s_id=199

Smoked cannabis therapy for HIV-related painful peripheral neuropathy
http://www.cannabis-med.org/studies…ow.php?s_id=172

Two cannabis based medicinal extracts for relief of central neuropathic pain
http://www.cannabis-med.org/studies…ow.php?s_id=143

Cannabis based medicinal extracts (CBME) in central neuropathic pain due to multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=82

Analgesic effect of the synthetic cannabinoid CT-3 on chronic neuropathic pain
http://www.cannabis-med.org/studies…how.php?s_id=85

Smoked cannabis in painful peripheral neuropathy and cancer pain refractory to opiods.
http://www.cannabis-med.org/studies…how.php?s_id=96

Analgesic effect of the cannabinoid analogue nabilone
http://www.cannabis-med.org/studies…ow.php?s_id=203

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Cannabis based treatments for neuropathic and multiple sclerosis-related pain.
http://www.unboundmedicine.com/medl…is_related_pain

Neuroprotectant
Marijuana Protects Your Brain
http://www.roninpub.com/art-mjbrain.html

The neuroprotective effect of cannabinoids in a rat model of Parkinson’s disease
http://www.medscape.com/medline/abstract/17196181

Neuroprotective and Intraocular Pressure-Lowering Effects of (-)Delta-THC
http://www.unboundmedicine.com/medl…del_of_Glaucoma

Neuroprotective effect of (-)Delta9-tetrahydrocannabinol and cannabidiol
http://www.unboundmedicine.com/medl…f_peroxynitrite

Neuroprotection induced by Delta(9)-tetrahydrocannabinol in AF5 cells
http://www.unboundmedicine.com/medl…ol_in_AF5_cells

Cannabidiol has a cerebroprotective action
http://www.unboundmedicine.com/medl…iting_mechanism

Cannabidiol but not Delta(9)-THC has a neuroprotective effect without the development of tolerance..
http://www.unboundmedicine.com/medl…nt_of_tolerance

Delta(9)-THC) prevents cerebral infarction
http://www.unboundmedicine.com/medl…ent_hypothermia

Delta(9)-Tetrahydrocannabinol protects hippocampal neurons from excitotoxicity
http://www.unboundmedicine.com/medl…_excitotoxicity

Cannabis and Neuroprotection
http://www.pacifier.com/~alive/cmu/…oprotection.htm

Medical marijuana uses – 700 medical marijuana clinical studies and papers

Nutrition
Oily fish makes ‘babies brainier’
http://news.bbc.co.uk/2/hi/health/4631006.stm

Efficacy of dietary hempseed oil in patients with atopic dermatitis.
http://www.medscape.com/medline/abs…ryText=hempseed

Effects of smoked marijuana on food intake and body weight
http://www.cannabis-med.org/studies…ow.php?s_id=117

Obesity
Does Cannabis Hold the Key to Treating Cardiometabolic Disease?
http://www.medscape.com/viewarticle/525040_print

Effects of smoked marijuana on food intake and body weight
http://www.cannabis-med.org/studies…ow.php?s_id=117

Osteoporosis
Prototype drug to prevent osteoporosis based on cannabinoids
http://www.news-medical.net/?id=15220

Hebrew U. Researchers Find Cannabis Can Strengthen Bones
http://www.israelnationalnews.com/News/News.aspx/96146

Peripheral cannabinoid receptor, CB2, regulates bone mass
http://www.pnas.org/cgi/content/abstract/103/3/696

New Weapon In Battle Against Osteoporosis
http://www.medicalnewstoday.com/articles/35621.php

Activation of CB2 receptor attenuates bone loss in osteoporosis
http://www.cannabis-med.org/english…el.php?id=210#2

Pain-
Cannabis effective at relieving pain after major surgery
http://www.news-medical.net/?id=17995

Cannabinoids, in combination with (NSAIDS), produce a synergistic analgesic effect
http://www.medjournal.com/forum/sho…587&postcount=1

Cannabinoids Among Most Promising Approaches to Treating Neuropathic Pain,
http://www.redorbit.com/news/health…source=r_health

Cannabinoid analgesia as a potential new therapeutic option
http://www.medscape.com/medline/abstract/16449552

Analgesic and adverse effects of an oral cannabis extract (Cannador) for postoperative pain
http://www.cannabis-med.org/studies…ow.php?s_id=184

Delta-9-THC based monotherapy in fibromyalgia patients
http://www.cannabis-med.org/studies…ow.php?s_id=194

Add-on treatment with the synthetic cannabinomimetic nabilone on patients with chronic pain -
http://www.cannabis-med.org/studies…ow.php?s_id=197

Nabilone significantly reduces spasticity-related pain
http://www.cannabis-med.org/studies…ow.php?s_id=200

Synergistic affective analgesic interaction between delta-9-tetrahydrocannabinol and morphine.
http://www.cannabis-med.org/studies…ow.php?s_id=178

Are oral cannabinoids safe and effective in refractory neuropathic pain?
http://www.cannabis-med.org/studies…ow.php?s_id=143

Dronabinol in the treatment of agitation in patients with Alzheimer’s disease with anorexia
http://www.cannabis-med.org/studies…how.php?s_id=61

Cannabis use for chronic non-cancer pain
http://www.cannabis-med.org/studies…how.php?s_id=91

Tetrahydrocannabinol for treatment of chronic pain
http://www.cannabis-med.org/studies…ow.php?s_id=147

Analgesic effect of the cannabinoid analogue nabilone
http://www.cannabis-med.org/studies…ow.php?s_id=203

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Pain relief with oral cannabinoids in familial Mediterranean fever.
http://www.cannabis-med.org/studies…how.php?s_id=18

The effect of orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

Marihuana as a therapeutic agent for muscle spasm or spasticity.
http://www.cannabis-med.org/studies…how.php?s_id=53

Analgesic effect of delta-9-tetrahydrocannabinol.
http://www.cannabis-med.org/studies…how.php?s_id=16

The analgesic properties of delta-9-tetrahydrocannabinol and codeine.
http://www.cannabis-med.org/studies…how.php?s_id=17

Most pain patients gain benefit from cannabis in a British study
http://www.cannabis-med.org/english…kel.php?id=84#1

Parkinson’s Disease
Marijuana Compounds May Aid Parkinson’s Disease
http://cannabisnews.com/news/19/thread19725.shtml

Marijuana-Like Chemicals Helps Treat Parkinson’s
http://cannabisnews.com/news/22/thread22608.shtml

Cannabis use in Parkinson’s disease: subjective improvement of motor symptoms.
http://www.cannabis-med.org/studies…how.php?s_id=33

Cannabinoids reduce levodopa-induced dyskinesia in Parkinson’s disease
http://www.cannabis-med.org/studies…how.php?s_id=54

Nabilone on L-DOPA induced dyskinesia in patients with idiopathic Parkinson’s disease
http://www.cannabis-med.org/studies…ow.php?s_id=153

Evaluation of cannabidiol in dystonic movement disorders.
http://www.cannabis-med.org/studies…how.php?s_id=14

Beneficial and adverse effects of cannabidiol in a Parkinson patient
http://www.cannabis-med.org/studies…ow.php?s_id=142

Neuroprotective effect of cannabinoids in a rat model of Parkinson’s disease
http://www.medscape.com/medline/abstract/17196181

Post Traumatic Stress Disorder
IDF TO TREAT SHELL SHOCK WITH CANNABIS
http://www.onlinepot.org/medical/id…sshellshock.htm

Study: Marijuana Eases Traumatic Memories
http://cannabisnews.com/news/13/thread13601.shtml

Medical Marijuana: PTSD Medical Malpractice
http://salem-news.com/articles/june…veque_61407.php

Cannabis for the Wounded – Another Walter Reed Scandal
http://www.libertypost.org/cgi-bin/…=179973&Disp=11

PTSD and Cannabis: A Clinician Ponders Mechanism of Action
http://ccrmg.org/journal/06spr/perspective2.html

Cannabis Eases Post Traumatic Stress
http://ccrmg.org/journal/06spr/ptsd.html

Endocannabinoids extinguish bad memories in the brain
http://www.cannabis-med.org/english…el.php?id=123#1

Natural high helps banish bad memories
http://www.newscientist.com/article…d-memories.html

Pregnancy
Oily fish makes ‘babies brainier’
http://news.bbc.co.uk/2/hi/health/4631006.stm

Ganja use among Jamaican women.
http://www.rism.org/isg/dlp/ganja/a…anjaBabyes.html

Dreher’s Jamaican Pregnancy Study
http://www.november.org/stayinfo/br…reherStudy.html

Cannabis Relieves Morning Sickness
http://ccrmg.org/journal/06spr/dreher.html#morning

Prenatal Marijuana Exposure and Neonatal Outcomes in Jamaica
http://www.druglibrary.org/Schaffer…/can-babies.htm

The Endocannabinoid-CB Receptor System
http://www.nel.edu/pdf_/25_12/NEL251204A01_Fride_.pdf

CLAIM #7: MARIJUANA USE DURING PREGNANCY HARMS THE FETUS
http://www.erowid.org/plants/cannab…bis_myth7.shtml

Prenatal exposure
Prenatal Marijuana Exposure and Neonatal Outcomes in Jamaica
http://www.druglibrary.org/Schaffer…/can-babies.htm

The Endocannabinoid-CB Receptor System
http://www.nel.edu/pdf_/25_12/NEL251204A01_Fride_.pdf

Ganja use among Jamaican women.
http://www.rism.org/isg/dlp/ganja/a…anjaBabyes.html

Dreher’s Jamaican Pregnancy Study
http://www.november.org/stayinfo/br…reherStudy.html

Nonmutagenic action of cannabinoids in vitro
http://trophort.com/005/993/005993433.html

Prenatal exposure to tobacco, alcohol, cannabis and caffeine on birth size and subsequent growth.
http://www.ncbi.nlm.nih.gov/sites/e…st_uids=3657756

Tobacco and marijuana use on offspring growth from birth through 3 years of age.
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Prenatal marijuana use and neonatal outcome.
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Pruritis
Cream with endocannabinoids effective in the treatment of pruritus
http://bbsnews.net/article.php/20051211212223236/print

Topical cannabinoid agonists : An effective new possibility for treating chronic pruritus.
http://www.cannabis-med.org/studies…ow.php?s_id=196

Dronabinol in patients with intractable pruritus secondary to cholestatic liver disease.
http://www.cannabis-med.org/studies…ow.php?s_id=116

Sativex
Sativex in the treatment of pain caused by rheumatoid arthritis
http://rheumatology.oxfordjournals….bstract/45/1/50

Sativex produced significant improvements in a subjective measure of spasticity
http://www.cannabis-med.org/studies…ow.php?s_id=170

Sativex in patients suffering from multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=168

Sativex in patients suffering from multiple sclerosis associated detrusor overactivity
http://www.cannabis-med.org/studies…ow.php?s_id=168

Sativex showed positive effects in 65 per cent of patients with chronic diseases
http://www.cannabis-med.org/english…el.php?id=230#4

Schizophrenia/ Mental disorders
Increased cannabinoid receptor density in the posterior cingulate cortex in schizophrenia.
http://www.medscape.com/medline/abstract/16710682

Symptoms of schizotypy precede cannabis use.
http://www.ukcia.org/forum/read.php?7,7543,7579

Cannabidiol as an antipsychotic
http://www.cannabis-med.org/studies…ow.php?s_id=171

Anandamide levels in cerebrospinal fluid of first-episode schizophrenic patients
http://www.unboundmedicine.com/medl…of_cannabis_use

Delta-9-Tetrahydrocannabinol-Induced Effects on Psychosis and Cognition
http://www.unboundmedicine.com/medl…s_and_Cognition

Cannabis is a First-Line Treatment for Childhood Mental Disorders
http://www.counterpunch.org/mikuriya07082006.html

Cannabis does not induce schizophrenia,
http://www.medicalnewstoday.com/articles/12283.php

Cannabis use does not cause schizophrenia
http://www.health.am/psy/more/canna…_schizophrenia/

Cannabinoids and psychosis.
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Cannabis as a psychotropic medication
http://bjp.rcpsych.org/cgi/content/full/185/1/78

Study Shows Long Term Marijuana Users Healthy
http://www.erowid.org/plants/cannab…_science3.shtml

Cannabis and schizophrenia link blurs further
http://www.newscientist.com/channel…rs-further.html

Evidence does not show a strong causal relation between the use of cannabis and psychosocial harm
http://www.library.nhs.uk/mentalHea…24106&tabID=289

——- Page 9

Sickle Cell Disease
Cannabis Relieves Sickle Cell Disease!
http://www.cannabisculture.com/foru…?Number=1155878

Sickle Cell Disease and Cannabis
http://www.pacifier.com/~alive/cmu/Sickle_cell.htm

Marijuana smoking in young adults with sickle cell
http://caribbean.scielo.org/scielo….&lng=en&nrm=iso

Medical use of cannabis in sickle cell disease
http://www.chanvre-info.ch/info/it/…-in-sickle.html

Cannabis use in sickle cell disease: a questionnaire study.
http://www.ncbi.nlm.nih.gov/sites/e…2&dopt=Abstract

Sleep modulation
Cannabidiol, a constituent of Cannabis sativa, modulates sleep in rats.
http://www.medscape.com/medline/abs…844117?prt=true

Dronabinol reduces signs and symptoms of idiopathic intracranial hypertension
http://www.cannabis-med.org/studies…ow.php?s_id=181

Cannabis-based medicine in central pain in multiple sclerosis.
http://www.cannabis-med.org/studies…ow.php?s_id=175

Two cannabis based medicinal extracts for relief of central neuropathic pain
http://www.cannabis-med.org/studies…how.php?s_id=15

Functional role for cannabinoids in respiratory stability during sleep
http://www.pacifier.com/~alive/cmu/…sleep_apnea.htm

THC reduces sleep apnoea in animal research
http://www.cannabis-med.org/english…el.php?id=120#1

Spasticity
The treatment of spasticity with Delta(9)-tetrahydrocannabinol in persons with spinal cord injury.
http://www.cannabis-med.org/studies…ow.php?s_id=166

Cannabis-based medicine in spasticity caused by multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=192

Cannabinoids in multiple sclerosis
http://www.cannabis-med.org/studies…ow.php?s_id=160

Sativex produced significant improvements in a subjective measure of spasticity
http://www.cannabis-med.org/studies…ow.php?s_id=170

Do cannabis-based medicinal extracts have general or specific effects on symptoms in ms?
http://www.cannabis-med.org/studies…how.php?s_id=56

Efficacy, safety and tolerability of an oral cannabis extract in the treatment of spasticity
http://www.cannabis-med.org/studies…how.php?s_id=63

Are oral cannabinoids safe and effective in refractory neuropathic pain?
http://www.cannabis-med.org/studies…ow.php?s_id=143

Experiences with THC-treatment in children and adolescents
http://www.cannabis-med.org/studies…how.php?s_id=80

The treatment of spasticity with D9-THC in patients with spinal cord injury
http://www.cannabis-med.org/studies…how.php?s_id=79

The effect of orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

Nabilone in the treatment of multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=11

Treatment of spasticity in spinal cord injury with dronabinol
http://www.cannabis-med.org/studies…ow.php?s_id=112

Delta-9-tetrahydrocannabinol shows antispastic and analgesic effects
http://www.cannabis-med.org/studies…how.php?s_id=10

Effect of cannabinoids on spasticity and ataxia in multiple sclerosis.
http://www.cannabis-med.org/studies…show.php?s_id=2

Delta-9-THC in the treatment of spasticity associated with multiple sclerosis.
http://www.cannabis-med.org/studies…show.php?s_id=1

Effect of Delta-9-THC on EMG Measurements in Human Spasticity
http://www.cannabis-med.org/studies…ow.php?s_id=110

The effect of delta-9-THC on human spasticity.
http://www.cannabis-med.org/studies…ow.php?s_id=154

Cannabis effect on spasticity in spinal cord injury.
http://www.cannabis-med.org/studies…ow.php?s_id=113

Treatment of human spasticity with delta 9-tetrahydrocannabinol.
` http://www.cannabis-med.org/studies…show.php?s_id=8

Marihuana as a therapeutic agent for muscle spasm or spasticity.
http://www.cannabis-med.org/studies…how.php?s_id=53

The perceived effects of marijuana on spinal cord injured males.
http://www.cannabis-med.org/studies…ow.php?s_id=138

Motor effects of delta 9 THC in cerebellar Lurcher mutant mice.
http://www.unboundmedicine.com/medl…her_mutant_mice

Cannabis-based medicine in spasticity caused by multiple sclerosis
http://www.unboundmedicine.com/medl…tiple_sclerosis

Spinal Cord Injury
The treatment of spasticity with Delta(9)-tetrahydrocannabinol in persons with spinal cord injury.
http://www.cannabis-med.org/studies…ow.php?s_id=166

Are oral cannabinoids safe and effective in refractory neuropathic pain?
http://www.cannabis-med.org/studies…ow.php?s_id=143

The treatment of spasticity with D9-THC) in patients with spinal cord injury
http://www.cannabis-med.org/studies…how.php?s_id=79

Delta-9-THC as an alternative therapy for overactive bladders in spinal cord injury
http://www.cannabis-med.org/studies…ow.php?s_id=102

The effect of orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

Treatment of spasticity in spinal cord injury with dronabinol
http://www.cannabis-med.org/studies…ow.php?s_id=112

Delta-9-tetrahydrocannabinol shows antispastic and analgesic effects
http://www.cannabis-med.org/studies…how.php?s_id=10

The effect of delta-9-THC on human spasticity.
http://www.cannabis-med.org/studies…ow.php?s_id=154

Cannabis effect on spasticity in spinal cord injury.
http://www.cannabis-med.org/studies…ow.php?s_id=113

Marihuana as a therapeutic agent for muscle spasm or spasticity.
http://www.cannabis-med.org/studies…how.php?s_id=53

The perceived effects of marijuana on spinal cord injured males.
http://www.cannabis-med.org/studies…ow.php?s_id=138

Stroke
Cannabidiol has a cerebroprotective action
http://www.unboundmedicine.com/medl…iting_mechanism

Delta(9)-THC) prevents cerebral infarction
http://www.unboundmedicine.com/medl…ent_hypothermia

Medical marijuana: study shows that THC slows atherosclerosis
http://thenexthurrah.typepad.com/th…al_marijua.html

Tea as medicine
Cannabis tea revisited: A systematic evaluation
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

THC/tetrahydrocannabinol
THC is effective in the treatment of tics in Tourette syndrome
http://www.cannabis-med.org/studies…how.php?s_id=98

THC effective in Tourette-Syndrome
http://www.pacifier.com/~alive/cmu/tourette_thc.htm

THC effective in Tourette syndrome in a 6-week trial
http://www.cannabis-med.org/english…el.php?id=146#1

Treatment of Tourette’s Syndrome With Delta-9-Tetrahydrocannabinol
http://ajp.psychiatryonline.org/cgi…/full/156/3/495

THC inhibits primary marker of Alzheimer’s disease
http://www.cannabis-med.org/english…el.php?id=225#3

THC improves appetite and reverses weight loss in AIDS patients
http://www.cannabis-med.org/studies…ow.php?s_id=189

Cancer-related anorexia-cachexia syndrome
http://www.unboundmedicine.com/medl…xia_Study_Group

THC effective in appetite and weight loss in severe lung disease (COPD)
http://www.cannabis-med.org/english…el.php?id=191#2

The antinociceptive effect of Delta9-tetrahydrocannabinol in the arthritic rat
http://www.unboundmedicine.com/medl…binoid_receptor

Synergy between Delta(9)-tetrahydrocannabinol and morphine in the arthritic rat
http://www.unboundmedicine.com/medl…e_arthritic_rat

Bronchial effects of aerosolized delta 9-tetrahydrocannabinol
http://www.cannabis-med.org/studies…ow.php?s_id=109

Bronchodilator effect of delta1-tetrahydrocannabinol administered by aerosol
http://www.cannabis-med.org/studies…how.php?s_id=60

Effects of smoked marijuana in experimentally induced asthma.
http://www.cannabis-med.org/studies…how.php?s_id=57

Marijuana and oral delta9-tetrahydrocannabinol on specific airway conductance
http://www.cannabis-med.org/studies…how.php?s_id=67

New Synthetic Delta-9-THC Inhaler Offers Safe, Rapid Delivery
http://www.medicalnewstoday.com/articles/22937.php

Smoked marijuana and oral delta-9-THC on specific airway conductance in asthmatic subjects
http://www.ukcia.org/research/Smoke…InAsthmatic.php

Delta(9)-tetrahydrocannabinol in patients with recurrent glioblastoma multiforme.
http://www.cannabis-med.org/studies…ow.php?s_id=193

9-Tetrahydrocannabinol Inhibits Cell Cycle Progression in Human Breast Cancer
http://cancerres.aacrjournals.org/c…ract/66/13/6615

THC and prochlorperazine effective in reducing vomiting in women following breast surgery
http://www.cannabis-med.org/english…el.php?id=219#1

{Delta}9-Tetrahydrocannabinol-Induced Apoptosis in Jurkat Leukemia T Cells
http://mcr.aacrjournals.org/cgi/con…bstract/4/8/549

Delta(9)-THC) prevents cerebral infarction
http://www.unboundmedicine.com/medl…ent_hypothermia

Medical marijuana: study shows that THC slows atherosclerosis
http://thenexthurrah.typepad.com/th…al_marijua.html

Delta-9-tetrahydrocannabinol shows antispastic and analgesic effects
http://www.cannabis-med.org/studies…how.php?s_id=10

The effect of delta-9-THC on human spasticity.
http://www.cannabis-med.org/studies…ow.php?s_id=154

The treatment of spasticity with D9-THC) in patients with spinal cord injury
http://www.cannabis-med.org/studies…how.php?s_id=79

Delta-9-THC as an alternative therapy for overactive bladders in spinal cord injury
http://www.cannabis-med.org/studies…ow.php?s_id=102

The effect of orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

The treatment of spasticity with Delta(9)-tetrahydrocannabinol in persons with spinal cord injury.
http://www.cannabis-med.org/studies…ow.php?s_id=166

Delta-9-Tetrahydrocannabinol-Induced Effects on Psychosis and Cognition
http://www.unboundmedicine.com/medl…s_and_Cognition

The effect of orally and rectally administered delta-9-tetrahydrocannabinol on spasticity
http://www.cannabis-med.org/studies…how.php?s_id=12

Marihuana as a therapeutic agent for muscle spasm or spasticity.
http://www.cannabis-med.org/studies…how.php?s_id=53

Analgesic effect of delta-9-tetrahydrocannabinol.
http://www.cannabis-med.org/studies…how.php?s_id=16

The analgesic properties of delta-9-tetrahydrocannabinol and codeine.
http://www.cannabis-med.org/studies…how.php?s_id=17

The perceived effects of smoked cannabis on patients with multiple sclerosis.
http://www.cannabis-med.org/studies…how.php?s_id=13

Cannabis use for chronic non-cancer pain
http://www.cannabis-med.org/studies…how.php?s_id=91

Tetrahydrocannabinol for treatment of chronic pain
http://www.cannabis-med.org/studies…ow.php?s_id=147

Delta-9-THC based monotherapy in fibromyalgia patients
http://www.cannabis-med.org/studies…ow.php?s_id=194

Delta(9)-THC) prevents cerebral infarction
http://www.unboundmedicine.com/medl…ent_hypothermia

Delta(9)-Tetrahydrocannabinol protects hippocampal neurons from excitotoxicity
http://www.unboundmedicine.com/medl…_excitotoxicity

Tobacco vs Cannabis-
Cannabis Smoke and Cancer: Assessing the Risk
http://www.norml.org/index.cfm?Group_ID=6891

Cannabis and tobacco smoke are not equally carcinogenic
http://www.pubmedcentral.nih.gov/ar…i?artid=1277837

Smoking Marijuana Does Not Cause Lung Cancer
http://www.mapinc.org/drugnews/v05/n1065/a03.html

Tobacco and marijuana use on offspring growth from birth through 3 years of age.
http://www.ncbi.nlm.nih.gov/sites/e…Pubmed_RVDocSum

Progression from marijuana use to daily smoking and nicotine dependence
http://www.erowid.org/references/refs_view.php?ID=6951

High anxieties – What the WHO doesn’t want you to know about cannabis
http://www.newscientist.com/article…t-cannabis.html

Radioactive tobacco
http://www.cannabisculture.com/news/tobacco/

Tourette’s Syndrome
Treatment of Tourette’s Syndrome With Delta-9-Tetrahydrocannabinol
http://ajp.psychiatryonline.org/cgi…/full/156/3/495

THC is effective in the treatment of tics in Tourette syndrome
http://www.cannabis-med.org/studies…how.php?s_id=98

Treatment of Tourette’s syndrome with Delta 9-tetrahydrocannabinol
http://www.cannabis-med.org/studies…how.php?s_id=99

Cannabinoids: possible role in patho-physiology and therapy of Gilles de la Tourette syndrome.
http://www.cannabis-med.org/studies…ow.php?s_id=100

THC effective in Tourette-Syndrome
http://www.pacifier.com/~alive/cmu/tourette_thc.htm

THC effective in Tourette syndrome in a 6-week trial
http://www.cannabis-med.org/english…el.php?id=146#1

Vaporizers
Vaporization as a smokeless cannabis delivery system
http://www.cannabis-med.org/studies…ow.php?s_id=187

Smokeless Cannabis Delivery Device Efficient And Less Toxic
http://www.medicalnewstoday.com/articles/71112.php

Volcano is to Vaporizer As Porsche is to Automobile
http://ccrmg.org/journal/04spr/volcano.html

Recommendation to Patients: “Don’t smoke, Vaporize”
http://ccrmg.org/journal/03sum/vaporize.html

Decreased respiratory symptoms in cannabis users who vaporize.
http://marijuana.researchtoday.net/archive/4/4/1195.htm

Use of vaporizers reduces toxins from cannabis smoke
http://www.cannabis-med.org/english…el.php?id=146#2

Wilson’s Disease
Cannabis sativa and dystonia secondary to Wilson’s disease.
http://www.medscape.com/medline/abstract/15390041

Cannabidiol Displays Antiepileptiform and Antiseizure Properties In Vitro and In Vivo

  • August 24, 2012 3:53 am
  1. Nicholas A. Jones,
  2. Andrew J. Hill,
  3. Imogen Smith,
  4. Sarah A. Bevan,
  5. Claire M. Williams,
  6. Benjamin J. Whalley and
  7. Gary J. Stephens

+ Author Affiliations


  1. School of Pharmacy (N.A.J., A.J.H., I.S., S.A.B., B.J.W., G.J.S.) and School of Psychology (N.A.J., A.J.H., C.M.W.), University of Reading, Whiteknights, Reading, United Kingdom
  1. Address correspondence to:
    Dr. Gary Stephens,
    School of Pharmacy, University of Reading, Whiteknights, P.O. Box 228, Reading RG6 6AJ, UK.

    E-mail: g.j.stephens@reading.ac.uk

Abstract

Plant-derived cannabinoids (phytocannabinoids) are compounds with emerging therapeutic potential. Early studies suggested that cannabidiol (CBD) has anticonvulsant properties in animal models and reduced seizure frequency in limited human trials. Here, we examine the antiepileptiform and antiseizure potential of CBD using in vitro electrophysiology and an in vivo animal seizure model, respectively. CBD (0.01–100 μM) effects were assessed in vitro using the Mg2+-free and 4-aminopyridine (4-AP) models of epileptiform activity in hippocampal brain slices via multielectrode array recordings. In the Mg2+-free model, CBD decreased epileptiform local field potential (LFP) burst amplitude [in CA1 and dentate gyrus (DG) regions] and burst duration (in all regions) and increased burst frequency (in all regions). In the 4-AP model, CBD decreased LFP burst amplitude (in CA1 only at 100 μM CBD), burst duration (in CA3 and DG), and burst frequency (in all regions). CBD (1, 10, and 100 mg/kg) effects were also examined in vivo using the pentylenetetrazole model of generalized seizures. CBD (100 mg/kg) exerted clear anticonvulsant effects with significant decreases in incidence of severe seizures and mortality compared with vehicle-treated animals. Finally, CBD acted with only low affinity at cannabinoid CB1 receptors and displayed no agonist activity in [35S]guanosine 5′-O-(3-thio)triphosphate assays in cortical membranes. These findings suggest that CBD acts, potentially in a CB1 receptor-independent manner, to inhibit epileptiform activity in vitro and seizure severity in vivo. Thus, we demonstrate the potential of CBD as a novel antiepileptic drug in the unmet clinical need associated with generalized seizures.

A growing number of phytocannabinoids have been shown to possess biological activity (Pertwee, 2008) and, in particular, to affect neuronal excitability in the CNS. Phytocannabinoid actions are reported to be mediated by G protein-coupled cannabinoid CB1 and CB2 receptors and potentially by other non-CB receptor targets (Howlett et al., 2004; Pertwee, 2008). CB1 receptors are highly expressed in the hippocampus (Herkenham et al., 1990; Tsou et al., 1998) and are well known to modulate epileptiform and seizure activity (Shen and Thayer, 1999; Wallace et al., 2001). Moreover, the endocannabinoid (eCB) system has been shown to be a key determinant of hippocampal epileptiform activity (Wallace et al., 2002; Monory et al., 2006; Ludányi et al., 2008). The major psychoactive compound Δ9-THC was the first phytocannabinoid reported to affect epileptiform activity; Δ9-THC, a partial agonist at CB1 receptors, was shown to inhibit excitatory glutamatergic neurotransmission in hippocampal neurons under low Mg2+ conditions (Shen and Thayer, 1999; but see Straiker and Mackie, 2005).

CBD is the major nonpsychoactive component of Cannabis sativa whose structure was first described by Mechoulam and Shvo (1963); CBD has recently attracted renewed interest for its therapeutic potential in a number of disease states (Pertwee, 2008). CBD has been proposed to possess anticonvulsive, neuroprotective, and anti-inflammatory properties in humans. Thus, within the CNS, CBD has been proposed to be protective against epilepsy, anxiety, and psychosis and to ameliorate diseases of the basal ganglia, such as parkinsonism and Huntington’s disease (Iuvone et al., 2009; Scuderi et al., 2009). CBD neuroprotective effects may be augmented by reported antioxidant properties (Hampson et al., 1998; Sagredo et al., 2007). Early studies suggested that CBD had anticonvulsant potential in one small-scale phase I clinical trial (Cunha et al., 1980). In this regard, there is a significant unmet clinical need for epilepsy, with ∼30% of epileptic patients experiencing intractable seizures regardless of conventional AED treatment (Kwan and Brodie, 2007). CBD is extremely well tolerated in humans; for example, CBD at doses of 600 mg does not precipitate any of the psychotic symptoms associated with Δ9-THC (Bhattacharyya et al., 2009). At present, CBD is used therapeutically in Sativex (1:1 Δ9-THC/CBD; GW Pharmaceuticals, Porton Down, UK) to alleviate pain symptoms in multiple sclerosis and cancer pain. CBD has anticonvulsant effects in animal models of maximal electroshock (Karler et al., 1974; Consroe and Wolkin, 1977; Consroe et al., 1982); however, CBD remains untested in other animal seizure models (Gordon and Devinsky, 2001) and so has yet to fulfill its potential indications as a clinical anticonvulsant.

In the present study, we demonstrate the potential of CBD as an AED. We show that CBD caused concentration-related and region-dependent attenuation of chemically induced epileptiform activity in hippocampal brain slices using in vitro MEA electrophysiological recordings. Furthermore, CBD reduced seizure severity and mortality in an in vivo model of generalized seizures. We also investigated the specific role of CB1 receptors in CBD action and found only a low-affinity interaction and lack of clear agonist effects. Overall, these data are consistent with CBD acting to mediate antiepileptiform and antiseizure effects in vitro and in vivo, respectively, potentially by CB1 receptor-independent mechanisms.

Materials and Methods

In Vitro Electrophysiology

Tissue Preparation and Solutions.

All experiments were performed in accordance with Home Office regulations [Animals (Scientific Procedures) Act 1986]. Acute transverse hippocampal brain slices (∼450 μm thick) were prepared from male and female (postnatal day ≥21) Wistar Kyoto rats using a Vibroslice 725M (Campden Instruments Ltd., Loughborough, Leicestershire, UK). Slices were produced and maintained in continuously carboxygenated (95% O2-5% CO2) artificial cerebrospinal fluid (aCSF) composed of 124 mM NaCl, 3 mM KCl, 1.25 mM KH2PO4, 1 mM MgSO4 · 6H2O, 36 mM NaHCO3, 2 mM CaCl2, and 10 mM d-glucose, pH 7.4. Spontaneous epileptiform activity was induced either by exchange of the standard aCSF perfusion media for aCSF with MgSO4 · 6H2O removed (Mg2+-free aCSF) or by addition of the K+ channel blocker 4-AP (100 μM; 4-AP aCSF).

MEA Electrophysiological Recording.

Substrate-integrated MEAs (Multi Channel Systems, Reutlingen, Germany) (Egert et al., 2002a; Stett et al., 2003) were used to record spontaneous neuronal activity as described previously (Ma et al., 2008). MEAs were composed of 60 electrodes (including reference ground) of 30 μm diameter, arranged in an ∼8 × 8 array with 200 μm spacing between electrodes.

MEAs were cleaned before each recording by immersion in 5% w/v Terg-A-Zyme (Cole-Palmer, London, UK) in distilled H2O, followed by methanol, and, finally, distilled H2O before air drying. Hippocampal sections immersed in aCSF were gently microdissected away from surrounding slice tissue using fine forceps under a WILD M8 binocular microscope (Leica AG, Solms, Germany). Dissected hippocampi were then adhered to the cleaned MEA surface using an applied and evaporated cellulose nitrate solution in methanol (∼4 μl, 0.24% w/v; Thermo Fisher Scientific, Leicestershire, UK) to ensure maximum contact between the tissue and recording electrodes and to avoid any physical stress on the tissue during recordings. Slices were observed at 4× magnification with a Nikon TS-51 inverted microscope (Nikon, Tokyo, Japan) and imaged via a Mikro-Okular camera (Bresser, Rhede, Germany) to map electrode positions to hippocampal regions. Slices were maintained at 25°C, continuously superfused (∼2 ml/min) with carboxygenated aCSF, and allowed to stabilize for at least 10 min before recordings. Signals were amplified (1200× gain), band pass-filtered (2–3200 Hz) by a 60-channel amplifier (MEA60 System, Multi Channel Systems), and simultaneously sampled at 10 kHz per channel on all 60 channels. Data were transferred to PC using MC_Rack software (Multi Channel Systems). Offline analysis of CBD effects upon burst amplitude, duration, and frequency was performed using MC_Rack, MATLAB 7.0.4. (Mathworks Inc., Natick, MA) and in-house analysis scripts. Animated contour plots of MEA-wide neuronal activity (Supplemental Fig. 1) were constructed from raw data files processed in MATLAB 6.5 using in-house code with functions adapted from MEA Tools and interpolated using a five-point Savitzky-Golay filter in MATLAB (Egert et al., 2002b). These data are displayed as peak source and peak sink animation frames. Burst propagation speeds were calculated by determining burst peak times at electrode positions closest to burst initiation (CA3) and termination (CA1) sites using MC_Rack and ImageJ software (Abramoff et al., 2004).

Data Presentation and Statistics.

Application of Mg2+-free and 4-AP aCSF induced spontaneous epileptiform activity characterized by recurrent status epilepticus-like local field potential (LFP) events (Figs. 2, A and B, and 4, A and B). We have recently characterized and validated the use of MEA technology to screen candidate AEDs in the Mg2+-free and 4-AP models using reference compounds, felbamate and phenobarbital (Hill et al., 2009). A discrete burst was defined as an LFP with both positive and negative components of greater than 2 S.D. from baseline noise. In each model, LFPs were abolished by the addition of the non-NMDA glutamate receptor antagonist 6-nitro-7-sulfamoylbenzo(f)quinoxaline-2–3-dione (5 μM) and tetrodotoxin (TTX, 1 μM) (n = 3 per model), indicating that epileptiform activity was due to firing of hippocampal neurons. After an initial 30-min control period, CBD was added cumulatively in increasing concentrations (30 min each concentration). Burst parameters (amplitude, duration, and frequency) were determined from the final 10 bursts of the control period or of each drug concentration. Increases in burst amplitude and decreases in frequency inherent to both in vitro models were observed over time in recordings in the absence of CBD (n = 4 per model). These changes required appropriate compensation to allow accurate assessment of CBD effects and have been rigorously modeled by us recently (Hill et al., 2009). Thus, burst frequency and amplitude from control recordings were normalized to the values observed after 30 min of epileptiform activity, then pooled to give mean values. Curves were fitted to resultant data, and derived equations were used to adjust values obtained from recordings in the presence of CBD. For amplitude; y = 0.8493 × e(x×−0.009295) + 0.4216 for Mg2+-free-induced bursting (r2 = 0.98) and y = 0.87 × e(−x/83.32) + 0.45 for 4-AP-induced bursting (r2 = 0.99), where x = time and y = burst amplitude. Frequency changes were more complex and required fifth-order polynomial equations: y = −9e−14x4 + 7e−10x3 − 3e−06x2 + 0.006x − 3.594 for Mg2+-free-induced bursting (r2 = 0.818); and y = 5−14x4 − 5e−10x3 + 2e−06x2 − 0.004x + 3.965 for 4-AP-induced bursting (r2 = 0.915), where x = time and y = frequency (Hill et al., 2009). Inevitable dead cell debris on the slice surface produced slice-to-slice variability in signal strength. Consequently, drug-induced changes are presented as changes to the stated measure versus control per experiment to provide normalized measures for pooled data. Statistical significance was determined by a nonparametric two-tailed Mann-Whitney U test. Mean propagation speeds (meters per second) were derived from pooled data and the significance of drug effects was tested using a two-tailed Student’s t test. In all cases, P ≤ 0.05 was considered significant.

Pharmacology.

The following agents were used: 6-nitro-7-sulfamoylbenzo(f)quinoxaline-2-3-dione (Tocris Cookson, Bristol, UK), tetrodotoxin (Alomone, Jerusalem, Israel), and 4-AP (Sigma-Aldrich, Poole, UK). CBD was kindly provided by GW Pharmaceuticals. CBD was made up as a 1000-fold stock solution in dimethylsulfoxide (Thermo Fisher Scientific, Leicestershire, UK) and stored at −20°C. Individual aliquots were thawed and dissolved in carboxygenated aCSF immediately before use. In all experiments, drugs were bath-applied (2 ml/min) for 30 min to achieve steady-state effects after the induction of epileptiform activity.

Pentylenetetrazole in Vivo Seizure Model

PTZ (80 mg/kg; Sigma-Aldrich) was used to induce seizures in 60 adult (postnatal day >21, 70–110 g) male Wistar Kyoto rats. In the days before seizure induction, animals were habituated to handling, experimental procedures, and the test environment. Before placement in their observation arenas, animals were injected intraperitoneally with CBD (1, 10, or 100 mg/kg); vehicle was a 1:1:18 solution of ethanol, Cremophor (Sigma-Aldrich), and 0.9% w/v NaCl. CBD is known to penetrate the blood-brain barrier such that 120 mg/kg delivered intraperitoneally in rats provides Cmax = 6.8 μg/g at Tmax = 120 min and, at the same dosage, no major toxicity, genotoxicity, or mutagenicity was observed (personal communication via GW Pharmaceuticals Ltd; Study Report UNA-REP-02). A group of animals that received volume-matched doses of vehicle alone served as a negative control. Sixty minutes after CBD or vehicle administration, animals were injected with 80 mg/kg PTZ i.p. to induce seizures. An observation system using closed-circuit television cameras (Farrimond et al., 2009) was used to monitor the behavior of up to five animals simultaneously from CBD/vehicle administration until 30 min after seizure induction. Input from closed-circuit TV cameras was managed and recorded by Zoneminder (version 1.2.3; Triornis Ltd., Bristol, UK) software and then was processed to yield complete videos for each animal.

Seizure Analysis.

Videos of PTZ-induced seizures were scored offline with a standard seizure severity scale appropriate for generalized seizures (Pohl and Mares, 1987) using Observer Video-Pro software (Noldus, Wageningen, The Netherlands). The seizure scoring scale was divided into stages as follows: 0, no change in behavior; 0.5, abnormal behavior (sniffing, excessive washing, and orientation); 1, isolated myoclonic jerks; 2, atypical clonic seizure; 3, fully developed bilateral forelimb clonus; 3.5, forelimb clonus with tonic component and body twist; 4, tonic-clonic seizure with suppressed tonic phase with loss of righting reflex; and 5, fully developed tonic-clonic seizure with loss of righting reflex (Pohl and Mares, 1987).

Specific markers of seizure behavior and development were assessed and compared between vehicle control and CBD groups. For each animal the latency (in seconds) from PTZ administration to the first sign of a seizure, development of a clonic seizure, development of tonic-clonic seizures, and severity of the seizure were recorded. In addition, the median severity, percentage of animals that experienced the highest seizure score (stage 5: tonic-clonic seizure), and percent mortality for each group were determined. Mean latency ± S.E.M. are presented for each group, together with the median value for seizure severity. Differences in latency and seizure duration values were assessed using one-way analysis of variance with a post hoc Tukey test; Mann-Whitney U tests were performed when replicant (n) numbers were insufficient to support post hoc testing. Differences in seizure incidence and mortality (percentage) were assessed by a nonparametric binomial test. In all cases, P ≤ 0.05 was considered to be significant.

Receptor Binding Assays

Membrane Preparation.

Cortical tissue was dissected from the brains of adult (postnatal day >21) male and female Wistar Kyoto rats and stored separately at −80°C until use. Tissue was suspended in a membrane buffer, containing 50 mM Tris-HCl, 5 mM MgCl2, 2 mM EDTA, and 0.5 mg/ml fatty acid-free bovine serum albumin (BSA) and complete protease inhibitor (Roche, Mannheim, Germany), pH 7.4, and was then homogenized using an Ultra-Turrax blender (Labo Moderne, Paris, France). Homogenates were centrifuged at 1000g at 4°C for 10 min, and supernatants were decanted and retained. Resulting pellets were rehomogenized and centrifugation was repeated as before. Supernatants were combined and then centrifuged at 39,00g at 4°C for 30 min in a high-speed Sorvall centrifuge; remaining pellets were resuspended in membrane buffer, and protein content was determined by the method of Lowry et al. (1951). All procedures were carried out on ice.

Radioligand Binding Assays.

Competition binding assays against the CB1 receptor antagonist [3H]SR141716A rimonabant were performed in triplicate in assay buffer containing 20 mM HEPES, 1 mM EDTA, 1 mM EGTA, and 5 mg/ml fatty acid-free BSA, pH 7.4. All stock solutions of drugs and membrane preparations were diluted in assay buffer and stored on ice immediately before incubation. Assay tubes contained 0.5 nM [3H]SR141716A (Kd = 0.53 ± 0.01 nM, n = 3, determined from saturation assay curves) together with drugs at the desired final concentration and were made up to a final volume of 1 ml with assay buffer. Nonspecific binding was determined in the presence of the CB1 receptor antagonist AM251 (10 μM). Assays were initiated by addition of 50 μg of membrane protein. Assay tubes were incubated for 90 min at 25°C, and the assay was terminated by rapid filtration through Whatman GF/C filters using a Brandell cell harvester, followed by three washes with ice-cold phosphate-buffered saline to remove unbound radioactivity. Filters were incubated overnight in 2 ml of scintillation fluid, and radioactivity was quantified by liquid scintillation spectrometry.

[35S]GTPγS Binding Assays.

Assays were performed in triplicate in assay buffer containing 20 mM HEPES, 3 mM MgCl2, 60 mM NaCl, 1 mM EGTA, and 0.5 mg/ml fatty acid-free BSA, pH 7.4. All stock solutions of drugs and membrane preparations were diluted in assay buffer and stored on ice immediately before use. Assay tubes contained GDP at a final concentration of 10 mM, together with drugs at the desired final concentration and were made up to a final volume of 1 ml with assay buffer. Assays were initiated by addition of 10 μg of membrane protein. Assays were incubated for 30 min at 30°C before addition of [35S]GTPγS (final concentration 0.1 nM). Assays were terminated after a further 30-min incubation at 30°C by rapid filtration through Whatman GF/C filters using a Brandell cell harvester, followed by three washes with ice-cold phosphate-buffered saline to remove unbound radioactivity. Filters were incubated for a minimum of 2 h in 2 ml of scintillation fluid, and radioactivity was quantified by liquid scintillation spectrometry.

Data Analysis and Statistical Procedures.

Data analyses were performed using GraphPad Prism (version 4.03; GraphPad Software Inc., San Diego, CA). Saturation experiments were performed to determine Kd and Bmax (picomoles per milligram) values; the free radioligand concentration was determined by subtraction of total bound radioligand from the added radioligand concentration. Data for specific radioligand binding and free radioligand concentration were fitted to equations describing one- or two-binding site models, and the best fit was determined using an F test. Saturation analyses best fit a one-binding site model. Competition experiments were fitted to one- and two-binding site models, and the best fit was determined using an F test. Data for the best fits are expressed as Ki values, with the respective percentage of high-affinity sites (percent Rh) given for two-binding site models (Vivo et al., 2006). The Hill slope for competition experiments was determined using a sigmoidal concentration-response model (variable slope). [35S]GTPγS concentration-response data were analyzed using a sigmoidal concentration-response model (variable slope) or linear regression and compared using an F test to select the appropriate model. No other constraints were applied. [35S]GTPγS binding is expressed as percentage increase in radioactivity as described previously (Dennis et al., 2008). All data are expressed as mean ± S.E.M.

Pharmacology.

The following agents were used: AM251, WIN55,212-2 (Tocris-Cookson, Bristol, UK), CBD (GW Pharmaceuticals), and [3H]SR141716A and [35S]GTPγS (GE Healthcare, Chalfont St. Giles, Buckinghamshire, UK). All other reagents were from Sigma-Aldrich.

Results

Characterization of Mg2+-Free and 4-AP Models of Epileptiform Activity Using MEA Electrophysiology.

To investigate neuronal excitability in vitro, we used both the Mg2+-free and 4-AP models of epileptiform activity in acute hippocampal brain slices, as measured using MEA electrophysiology. Two separate models of epileptiform activity were used to provide a broader analysis of drug effects (Hill et al., 2009; Whalley et al., 2009). Hippocampal slices have a well defined architecture (Fig. 1A), exhibited no spontaneous LFP events in control aCSF, and proved readily amenable to MEA recording (Fig. 1B). We sought to take advantage of the ability of MEAs to record spatiotemporal activity at multiple discrete, identifiable regions by investigating activity at CA1, CA3, and DG regions within the hippocampus. Application of Mg2+-free aCSF (Fig. 1C) or 4-AP aCSF (Fig. 1D) to hippocampal slices resulted in the appearance of robust spontaneous epileptiform LFPs across the preparation. LFPs were consistent with status epilepticus-like activity and were reliably recorded using the multisite MEA technique (Table 1). Slice-to-slice variability and electrode contact variability resulted in substantial variation in signal strength (Table 1); therefore, subsequent drug-induced changes in burst characteristics were normalized to control bursts before drug application (these analyses are fully characterized in Hill et al., 2009).

Fig. 1.

View larger version:

Fig. 1.

Hippocampal slices are amenable to MEA recording. A, schematic representation of hippocampal slice showing the position of CA1, CA3, and DG regions, together with major pathways: Schaffer collateral (SC), mossy fiber (MF), and perforant pathway (PP). B, micrograph showing a hippocampal brain slice (stained with pontamine blue) mounted onto a substrate-integrated MEA (60 electrodes of 30 μm diameter, spaced 200 μm apart in an ∼8 × 8 arrangement). Scale bar, 400 μm. Representative LFP burst activity was recorded at 60 electrodes across a hippocampal slice in (C) Mg2+-free aCSF and (D) 4-AP aCSF. Traces were high pass-filtered in an MC_rack at 2 Hz.

Table 1

Characterization measures for the Mg2+-free and 4-AP (100 μM) aCSF-induced LFP epileptiform activity in the CA1, CA3, and DG regions of the hippocampus

LFP peak burst amplitude values are presented as a minimum and maximum range and mean ± S.E.M. LFP burst duration and frequencies are presented as mean ± S.E.M. A minimum of six separate hippocampal slices were used in the characterisation of each in vitro model.

Effects of CBD in the Mg2+-Free Model of in Vitro Epileptiform Activity.

We first examined the effects of CBD in the Mg2+-free model to assess CBD effects on a receptor-dependent model of epileptiform activity. The Mg2+-free model removes the Mg2+-dependent block of NMDA glutamate receptors, rendering them more responsive to synaptically released glutamate at resting membrane potentials. In Mg2+-free aCSF, CBD significantly decreased LFP burst amplitude in the CA1 (1–100 μM CBD) and DG (10–100 μM CBD) regions (Fig. 2, A, B, and Ci). In contrast, CBD (0.01–100 μM) effects on LFP burst amplitude in CA3 failed to reach significance. CBD decreased burst duration in CA1 (0.01–100 μM CBD), CA3 (0.01–100 μM CBD), and DG (0.1–100 μM CBD) regions (Fig. 2, A, B, and Cii). CBD (0.01–10 μM) also caused an increase in burst frequency in all regions tested (Fig. 2Ciii; however, this effect was lost at 100 μM CBD. To correlate these data with information on LFP burst initiation and spread across the hippocampal brain slice, we constructed contour plots (Fig. 3A) and associated video animations (Supplemental Fig. 1). Such plots spatiotemporally visualize the “8 × 8” MEA configuration (Fig. 1, B and C) and the individual LFP activity shown in raw data traces (Fig. 2, A and B). In these experiments, Mg2+-free aCSF-induced bursts typically originated in the CA3 region of the hippocampal slice preparation and propagated along the principal cell layer toward CA1. LFP events induced by Mg2+-free aCSF had a mean propagation speed of 0.229 ± 0.048 m/s (n = 6). CBD (100 μM) caused a clear suppression of Mg2+-free-induced LFP burst amplitude peak source and peak sink values across the hippocampal slice (Fig. 3A; Supplemental Fig. 1). Propagation speed across the brain slice in Mg2+-free aCSF was not affected by 100 μM CBD (0.232 ± 0.076 m/s; n = 6; P > 0.5). Taken together, these data suggest that, although CBD attenuates epileptiform LFP amplitude and duration in the Mg2+-free model, the rate of signal spread across the preparation is not changed (see Discussion).

Fig. 2.

View larger version:

Fig. 2.

CBD attenuates epileptiform activity induced by Mg2+-free aCSF. A, representative traces showing the effects of 100 μM CBD on Mg2+-free aCSF-induced LFP bursts in different regions of hippocampal slices. Dotted lines represent an individual LFP (as shown in B). B, effects of 1 and 100 μM CBD on a representative individual Mg2+-free aCSF-induced LFP burst. C, bar graphs showing the effects of acute treatment of increasing CBD concentrations on normalized burst amplitude (Ci), normalized burst duration (Cii), and normalized burst frequency in Mg2+-free aCSF (Ciii). Note that burst amplitudes have been adjusted for run-down and burst frequencies have been adjusted for run-up as described under Materials and Methods. Values are means ± S.E.M. for the last 10 LFP bursts in each condition. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001 (two-tailed Mann-Whitney U test).

Fig. 3.

View larger version:

Fig. 3.

CBD attenuates epileptiform activity induced by Mg2+-free and 4-AP aCSF. Representative contour plots illustrating CBD effects upon spatiotemporal epileptiform burst features. A, in the continued presence of Mg2+-free aCSF: quiescent period between epileptiform burst events also showing hippocampal slice orientation (i), peak source in the absence of CBD (ii), and peak source in the presence of CBD (100 μM) (iii). B, in the continued presence of 100 μM 4-AP: quiescent period between epileptiform burst events also showing hippocampal slice orientation (i), peak source in the absence of CBD (ii), and peak source in the presence of CBD (100 μM) (iii).

Effects of CBD in the 4-AP Model of in Vitro Epileptiform Activity.

We next examined the effects of CBD on epileptiform bursting events in the 4-AP model of status epilepticus-like activity. 4-AP acts to block postsynaptic voltage-dependent K+ channels and inhibits neuronal repolarization to effectively increase excitability. In 100 μM 4-AP aCSF, CBD (100 μM) caused a significant decrease in LFP burst amplitude in CA1 only (Fig. 4, A, B, and Ci). In contrast, CBD (0.01–0.1 μM) caused an unexpected small, but significant, increase in LFP burst amplitude in the DG, which was not apparent at higher CBD concentrations in this or other hippocampal regions (Fig. 4Ci). CBD caused a decrease in burst duration in the DG (0.01–100 μM CBD) and CA3 (0.1–100 μM CBD) but was without an overall effect on CA1 (Fig. 4, A, B, and Cii). CBD (0.01–100 μM) also caused a significant decrease in burst frequency in all regions tested (Fig. 4Ciii). In the same manner as for the Mg2+-free model, contour plots of 4-AP-induced epileptiform LFP burst events (Fig. 3B) permitted spatiotemporal visualization of activity across the slice preparation (Supplemental Fig. 1). 4-AP aCSF-induced bursts typically were initiated in CA3 before spreading to CA1 with a propagation speed of 0.146 ± 0.033 m/s (n = 5). CBD (100 μM) caused a clear suppression of 4-AP-induced epileptiform LFP burst amplitude (Fig. 3B; Supplemental Fig. 1). Propagation speed across the brain slice in 4-AP aCSF was not affected by 100 μM CBD (0.176 ± 0.046 m/s, n = 6; P > 0.5).

Fig. 4.

View larger version:

Fig. 4.

CBD attenuates epileptiform activity induced by 4-AP aCSF. A, representative traces showing effects of 100 μM CBD on 4-AP aCSF-induced LFP bursts in different regions of a hippocampal slice. Dotted lines represent an individual LFP (as shown in B). B, effects of 1 and 100 μM CBD on a representative individual 4-AP aCSF-induced LFP burst. C, bar graphs showing the effects of acute treatment of increasing CBD concentrations on normalized burst amplitude (Ci), normalized burst duration (Cii), and normalized burst frequency in the 4-AP aCSF (Ciii). Note that burst amplitudes have been adjusted for run-down and burst frequencies have been adjusted for run-up as described under Materials and Methods. Values are means ± S.E.M. for the last 10 LFP bursts in each condition. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001 (two-tailed Mann-Whitney U test).

Taken together, these data show that CBD displayed clear concentration-related, region-specific, anticonvulsant properties in two different in vitro models of epileptiform activity, attenuating LFP burst amplitude and duration, but with no effect on the rate of signal propagation in either model.

Effects of CBD in the PTZ Model of Generalized Seizures.

We next assessed the effects of CBD (1, 10, and 100 mg/kg i.p.) on PTZ-induced generalized seizures in adult male rats. PTZ acts as a GABAA receptor antagonist and this model is well defined and used as a standard for the identification of potential anticonvulsants to treat generalized clonic seizures (Löscher et al., 1991). Seizures were defined by a standard scoring scale (see Materials and Methods). CBD at any dose did not significantly alter the latency to the first sign of PTZ-induced seizures (Fig. 5A) or latency to development of clonic (Fig. 5B) seizures. Unexpectedly, CBD (1 mg/kg) reduced latency to tonic-clonic seizures (P < 0.01) (Fig. 5C). No other effects of CBD on latency to specific seizure states were observed. In contrast to the lack of definitive effects on seizure latency, CBD (100 mg/kg) demonstrated clear anticonvulsant effects via measures of seizure severity (Fig. 6, A and B) and mortality (Fig. 6C). When the severity of PTZ-induced seizures is considered, vehicle-treated animals reached a median score of 5 (tonic-clonic seizures with a loss of righting reflex), the most severe on the scoring scale (Fig. 6A). In contrast, animals treated with 100 mg/kg CBD exhibited a significantly reduced median score of 3.5 (forelimb clonus with a tonic component, but with the righting reflex preserved; n = 15 animals; P < 0.001) (Fig. 6A). This was associated with a marked decrease in the proportion of animals that developed the most severe tonic-clonic seizures, which was reduced from 53% in vehicle to 7% by 100 mg/kg CBD (n = 15 animals, P < 0.001) (Fig. 6B). Finally, percent mortality was significantly reduced from 47% in vehicle to 7% by 100 mg/kg CBD (n = 15 animals, P < 0.001) (Fig. 6C). Overall, these in vivo data confirm our in vitro results above and fully support an anticonvulsant action for CBD.

Fig. 5.

View larger version:

Fig. 5.

CBD has no clear effects on seizure latency in vivo. Bar graphs showing lack of effects of CBD (1, 10, and 100 mg/kg) on latency to the first sign of a seizure (A), latency to clonic seizures (B), and latency to tonic-clonic seizures (C). Each data set n = 15 animals. Note that CBD (1 mg/kg) reduced latency to tonic-clonic seizures; this proconvulsant action was not observed at higher CBD doses. **, P ≤ 0.01 (one-way analysis of variance) with a post hoc Tukey test).

Fig. 6.

View larger version:

Fig. 6.

CBD reduces seizure severity and mortality in vivo. Bar graphs showing effects of CBD (1, 10, and 100 mg/kg) on median seizure severity (A), percentage of animals reaching tonic-clonic seizures (B), and percent mortality (C). Each data set n = 15 animals. CBD (100 mg/kg) significantly reduced all of these parameters: ***, P ≤ 0.001 (nonparametric binomial test).

Effects of CBD in Receptor Binding Assays.

It is known that hippocampal CB1 receptor expression on glutamatergic terminals is selectively down-regulated under epileptic conditions (Ludányi et al., 2008); moreover, activation of CB1 receptors by eCBs is protective against seizures (Monory et al., 2006) and exogenous CB1 agonists decrease epileptiform activity in hippocampal neurons (Shen and Thayer, 1999; Blair et al., 2006). Therefore, we determined potential CBD actions at CB1 receptors. Competition binding assays were performed for CBD against the CB1 receptor antagonist [3H]SR141716A in isolated cortical membranes; CBD effects were compared with those of the standard synthetic CB receptor agonist WIN55,212-2 and the CB1 receptor antagonist AM251 (Fig. 7A). AM251 displacement of [3H]SR141716A binding occurred with high affinity (Ki = 190 ± 56 pM; n = 4) and was best fitted by a one-site competition model (Hill slope = −1.08 ± 0.13; n = 4). In contrast, WIN55,212-2 displacement was best fitted to a two-site model with a high-affinity site (Ki = 7.03 ± 4.1 nM; % Rh = 27.4 ± 5.0%; n = 4) and a low-affinity site (Ki = 904 ± 155 nM; n = 4); in these experiments, Hill slopes for either the low- or high-affinity site did not match unity. CBD displacement of [3H]SR141716A occurred with low affinity (Ki = 1.82 ± 0.38 μM; n = 4) and was best fitted by a one-site model (Hill slope = −1.15 ± 0.11; n = 4).

Fig. 7.

View larger version:

Fig. 7.

CBD displaces [3H]SR141716A binding with low affinity and lacks agonist effects in [35S]GTPγS binding assays in cortical membranes. A, representative competition curves for the CB receptor agonist WIN55,212-2, the selective CB1 receptor antagonist AM251, and CBD against 1 nM [3H]SR141716A (a selective CB1 receptor antagonist) binding to cortical membranes. Points are means ± S.E.M. of triplicate points. B, agonist-binding curves for the CB receptor agonist WIN55,212-2, the selective CB1 receptor antagonist AM251, and CBD stimulation of [35S]GTPγS binding to cortical membranes. Points are means ± S.E.M. of triplicate points from three separate experiments.

Finally, we investigated potential functional effects of CBD using [35S]GTPγS binding assays in rat cortical membranes; CBD actions were compared with effects of WIN55,212-2 and AM251 (Fig. 7B). We first confirmed the presence of functional CB receptors. Accordingly, WIN55,212-2 caused an increase in percent stimulation of [35S]GTPγS binding with an EC50 of 95.1 ± 0.1 nM (n = 3); for 10 μM WIN55,212-2, Emax = 98.6 ± 3.5% (n = 3). AM251 had no stimulatory effect on [35S]GTPγS binding at tested concentrations of <1 μM; at micromolar concentrations AM251 caused a moderate depression of [35S]GTPγS binding (10 μM AM251: −20.3 ± 4.3%, n = 3). CBD had no effect at concentrations ≤1 μM; large decreases in [35S]GTPγS binding were seen at 10 μM CBD (−28.8 ± 10.3%, n = 3) and 100 μM CBD (−76.7 ± 15.9%, n = 3).

Thus, overall, CBD had clear antiepileptogenic and antiseizure effects but only low affinity and no clear agonist effects at cortical CB1 receptors.

Discussion

CBD Reduces Excitability in in Vitro Models of Epileptiform Activity.

In the present study, we use extracellular MEA recordings to demonstrate that CBD attenuates epileptiform activity in both the Mg2+-free and 4-AP in vitro models of status epilepticus in the mammalian hippocampus, a prominent epileptogenic region (Ben-Ari and Cossart, 2000). The major effects of CBD were to decrease LFP burst amplitude and duration in a hippocampal region-specific manner. In general, the CA1 region was most sensitive to CBD effects. Thus, LFP amplitude was significantly reduced at lower CBD concentrations in CA1 than in CA3 (with DG remaining unaffected) in Mg2+-free aCSF, and CA1 was the only region in which LFP amplitude was affected in 4-AP aCSF. This is of interest because the CA1 region represents the major output of the hippocampus, relaying information to cortical and subcortical sites and is intimately involved in propagation of epileptic activity (McCormick and Contreras, 2001). Contour plots constructed from data in the Mg2+-free and 4-AP models confirmed that LFP bursts typically originated in the CA3 region and propagated toward CA1 (Feng and Durand, 2005), strongly suggesting that CA1 is a major focus of epileptic activity in the two models used and illustrating that CBD exerts a significant antiepileptiform effect in this region.

Overall, CBD induced more prominent effects in Mg2+-free than in 4-AP aCSF. This result may reflect inherent differences between the two models, which affect NMDA glutamate receptors and K+ channels, respectively. CBD had contrasting actions on LFP burst frequency between models; frequency was increased in all regions by CBD in Mg2+-free aCSF but was decreased in all regions in 4-AP aCSF. It is interesting to note that 100 μM CBD was without effect on burst frequency in the Mg2+-free model, in contrast to data for all the lower concentrations of CBD tested. This finding was the only indication of any biphasic action of CBD, a common phenomenon associated with cannabinoids whereby increasing concentrations cause changes in the pharmacological “direction” of action (Pertwee, 2008).

In light of the region-specific effects of CBD, it will be of interest in the future to investigate the cellular mechanisms of action of CBD using intracellular recording from individual neurons in selected hippocampal regions. In both the Mg2+-free and 4-AP models, contour plots and subsequent analyses showed that CBD caused clear attenuation in LFP burst amplitude but had no overall effect on burst propagation speed. These findings suggest that CBD acts to reduce the magnitude of epileptiform activity while leaving speed of information transmission across the hippocampal slice intact. It is possible that this action may result in a more tolerable side effect profile for CBD in comparison with existing AEDs if used in a clinical setting.

CBD Has Anticonvulsant Properties in the PTZ Model of Generalized Seizures.

CBD had beneficial effects on seizure severity and lethality in response to PTZ administration without delaying the time taken for seizures to develop. CBD (100 mg/kg) demonstrated clear anticonvulsant effects in terms of significant reductions in median seizure severity, tonic-clonic seizures, and mortality. Particularly striking effects were that <10% of animals developed tonic-clonic seizures or died when treated with CBD in comparison to approximately 50% of vehicle-treated animals. The present data strongly substantiate a number of earlier in vivo studies suggesting that CBD has anticonvulsant potential (Lutz, 2004; Scuderi et al., 2009). CBD has been reported to have relatively potent anticonvulsant action in maximal electroshock (a model of partial seizure with secondary generalization) (Karler et al., 1974; Consroe and Wolkin, 1977). Moreover, CBD prevented tonic-clonic seizures in response to electroshock current (Consroe et al., 1982). There are limited clinical data on CBD effects on seizure frequency in humans (Gordon and Devinsky, 2001). However, in one small double-blind study of eight patients with uncontrolled secondary generalized epilepsy treated with 200 to 300 mg of CBD, four remained symptom-free and three had signs of improvement (Cunha et al., 1980). One potential concern was the high doses of CBD used by Cunha et al. (1980). Because all new therapies must be introduced initially in an adjunct capacity to existing medication, the present study suggests that one attractive possibility is a role for CBD as an adjunct in generalized seizures. In this regard, earlier animal studies indicate that CBD enhances the effects of phenytoin (although CBD reduced the potency of other AEDs) (Consroe and Wolkin, 1977). In the future, it will be of interest to extend studies to other animal seizure models and also to combination therapies with selective AEDs to determine the full clinical anticonvulsant potential of CBD against a range of epilepsy phenotypes.

Mechanism of Action.

Cannabinoid actions are mediated by CB1 and CB2 receptors, potentially by the GPR55 receptor, and also by cannabinoid receptor-independent mechanisms (Howlett et al., 2004; Ryberg et al., 2007). In regard to epilepsy, CB1 receptors are densely expressed in the hippocampal formation (Herkenham et al., 1990; Tsou et al., 1998) where their activation is widely reported to be antiepileptic in animal models (Shen and Thayer, 1999; Wallace et al., 2001; but see Clement et al., 2003). Here, we demonstrate that CBD displaced the selective CB1 receptor antagonist [3H]SR141716A in cortical membranes with relatively low affinity (Ki = 1.82 μM); these data are in line with values reported in whole brain membranes (reviewed in Pertwee, 2008) and our data in cerebellar membranes (Smith et al., 2009). CB receptor/G-protein coupling may differ among distinct brain regions (Breivogel et al., 1997; Dennis et al., 2008); therefore, we investigated CBD effects on [35S]GTPγS binding in isolated cortical membranes. We showed that CBD has no stimulatory agonist activity but that CBD at micromolar concentrations decreases G protein activity. These findings are also in agreement with studies in mouse whole brain membranes (Thomas et al., 2007), which showed that CBD has only low affinity at CB1 and CB2 receptors but acts efficaciously as an antagonist at both receptor types (Thomas et al., 2007; see Pertwee, 2008). There are a number of potential mechanisms by which ligands acting at CB receptors may mediate anticonvulsant effects. Receptor agonists may act at CB1 on excitatory presynaptic terminals to inhibit glutamate neurotransmitter release. Such a mechanism is unlikely here as CBD has no agonist effect in GTPγS binding assays (Thomas et al., 2007). An alternative is that antagonists act at CB1 receptors on inhibitory presynaptic terminals to increase GABA release. We have demonstrated such a mechanism for the phytocannabinoid Δ9-tetrahydrocannabivarin in the cerebellum (Ma et al., 2008), where displacement of eCB tone may lead to increased inhibition. It may also be speculated that the decreases in G protein activity seen in GTPγS binding assays represent an inverse agonist action at CB1 receptors; for example, if CBD were acting as an inverse agonist at CB1 receptors on inhibitory presynaptic terminals an increase in GABA release could lead to reduced excitability. However, mechanisms involving increases in GABA release are unlikely here as CBD was effective in reducing seizures in vivo in the presence of the GABAA receptor antagonist PTZ. Moreover, CBD-induced reductions in [35S]GTPγS binding to whole brain membranes were retained in CB1 knockout [cnr1(−/−)] mice, suggesting that CBD is not an inverse agonist at CB1 receptors (Thomas et al., 2007). Overall, the low affinity and lack of agonist activity at CB1 receptors suggests that the CBD anticonvulsant effects reported here are potentially mediated by CB1 receptor-independent mechanisms. In addition to the study of Thomas et al., showing that CBD actions were unaltered in cnr1(−/−) mice, CBD anticonvulsant effects in the maximal electroshock model were unaffected by the CB1 receptor antagonist SR141716A, whereas those of Δ9-THC and WIN55,212-2 were blocked (Wallace et al., 2001).

In addition to CB receptors, a number of alternative molecular targets may also contribute to CBD effects on neuronal excitability. CBD has been reported to be an antagonist at GPR55, a non-CB1/CB2 receptor (Ryberg et al., 2007); in contrast, a recent study demonstrated that CBD has no effect at GPR55 (Kapur et al., 2009). CBD may cause an increase in anticonvulsant eCBs via the reported inhibition of the catabolic enzyme fatty acid hydrolyase, which degrades anandamide, and/or the blockade of anandamide uptake (Watanabe et al., 1996; Rakhshan et al., 2000; Bisogno et al., 2001). CBD is reported to be a weak agonist at human TRPV1 receptors (Bisogno et al., 2001); a more recent study suggests an action for CBD at rat and human transient receptor potential vanilloid 2 but not rat transient receptor potential vanilloid 1 receptors (Qin et al., 2008). More relevant to potential effects on neuronal excitability in the CNS is the demonstration that CBD exerts a bidirectional action on [Ca2+]i levels in hippocampal neurons (Ryan et al., 2009). Under control conditions, CBD induces increases in [Ca2+]i; in contrast, in the presence of 4-AP (which induces seizure-like [Ca2+]i oscillations) or increased extracellular K+, CBD acts to reduce [Ca2+]i and thus epileptiform activity, via an action on mitochondria Ca2+ stores. A further recent report provides the first evidence that CBD can also block low-voltage-activated (T-type) Ca2+ channels (Ross et al., 2008), important modulators of neuronal excitability. Finally, CBD may also enhance the activity of inhibitory glycine receptors (Ahrens et al., 2009). Overall, the demonstration that CBD acts on multiple molecular targets that each play a key role in neuronal excitability reinforces the potential of CBD as an AED.

In conclusion, our data in separate in vitro models of epileptiform activity and, in particular, the beneficial reductions in seizure severity caused by CBD in an in vivo animal model of generalized seizures suggests that earlier, small-scale clinical trials for CBD in untreated epilepsy warrant urgent renewed investigation.

Acknowledgments

We thank Professor Philip Strange for useful discussion and Colin Stott (GW Pharmaceuticals) and Professor Gernot Riedel (University of Aberdeen) for pharmacokinetics data.

Footnotes

  • This work was supported by a GW Pharmaceuticals and Otsuka Pharmaceuticals award, by a University of Reading Research Endowment Trust Fund award; and The Wellcome Trust [Grant 070739].

  • Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.

    doi:10.1124/jpet.109.159145

  • Graphic The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.

  • ABBREVIATIONS:

    CNS
    central nervous system
    CB
    cannabinoid
    eCB
    endocannabinoid
    Δ9-THC
    Δ9-tetrahydrocannabinol
    CBD
    cannabidiol
    AED
    antiepileptic drug
    MEA
    multielectrode array
    aCSF
    artificial cerebrospinal fluid
    4-AP
    4-aminopyridine
    LFP
    local field potential
    NMDA
    N-methyl-d-aspartate
    BSA
    bovine serum albumin
    SR141716A
    N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide
    AM251
    N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide
    GTPγS
    guanosine 5′-O-(3-thio)triphosphate
    WIN55,212-2
    [2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-napthalenylmethanone
    PTZ
    pentylenetetrazole.
    • Received July 22, 2009.
    • Accepted November 9, 2009.

References

    1. Abramoff MD,
    2. Magelhaes PJ,
    3. Ram SJ

    (2004) Image processing with ImageJ. J Biophoton Int 11:36–42.

    1. Ahrens J,
    2. Demir R,
    3. Leuwer M,
    4. de la Roche J,
    5. Krampfl K,
    6. Foadi N,
    7. Karst M,
    8. Haeseler G

    (2009) The nonpsychotropic cannabinoid cannabidiol modulates and directly activates α1 and α1β glycine receptor function. Pharmacology 83:217–222.

    1. Ben-Ari Y,
    2. Cossart R

    (2000) Kainate, a double agent that generates seizures: two decades of progress. Trends Neurosci 23:580–587.

    1. Bhattacharyya S,
    2. Fusar-Poli P,
    3. Borgwardt S,
    4. Martin-Santos R,
    5. Nosarti C,
    6. O’Carroll C,
    7. Allen P,
    8. Seal ML,
    9. Fletcher PC,
    10. Crippa JA,
    11. et al

    . (2009) Modulation of mediotemporal and ventrostriatal function in humans by Δ9-tetrahydrocannabinol: a neural basis for the effects of Cannabis sativa on learning and psychosis. Arch Gen Psychiatry 66:442–451.

    1. Bisogno T,
    2. Hanus L,
    3. De Petrocellis L,
    4. Tchilibon S,
    5. Ponde DE,
    6. Brandi I,
    7. Moriello AS,
    8. Davis JB,
    9. Mechoulam R,
    10. Di Marzo V

    (2001) Molecular targets for cannabidiol and its synthetic analogues: effect on vanilloid VR1 receptors and on the cellular uptake and enzymatic hydrolysis of anandamide. Br J Pharmacol 134:845–852.

    1. Blair RE,
    2. Deshpande LS,
    3. Sombati S,
    4. Falenski KW,
    5. Martin BR,
    6. DeLorenzo RJ

    (2006) Activation of the cannabinoid type-1 receptor mediates the anticonvulsant properties of cannabinoids in the hippocampal neuronal culture models of acquired epilepsy and status epilepticus. J Pharmacol Exp Ther 317:1072–1078.

    1. Breivogel CS,
    2. Sim LJ,
    3. Childers SR

    (1997) Regional differences in cannabinoid receptor/G-protein coupling in rat brain. J Pharmacol Exp Ther 282:1632–1642.

    1. Clement AB,
    2. Hawkins EG,
    3. Lichtman AH,
    4. Cravatt BF

    (2003) Increased seizure susceptibility and proconvulsant activity of anandamide in mice lacking fatty acid amide hydrolase. J Neurosci 23:3916–3923.

    1. Consroe P,
    2. Benedito MA,
    3. Leite JR,
    4. Carlini EA,
    5. Mechoulam R

    (1982) Effects of cannabidiol on behavioral seizures caused by convulsant drugs or current in mice. Eur J Pharmacol 83:293–298.

    1. Consroe P,
    2. Wolkin A

    (1977) Cannabidiol-antiepileptic drug comparisons and interactions in experimentally induced seizures in rats. J Pharmacol Exp Ther 201:26–32.

    1. Cunha JM,
    2. Carlini EA,
    3. Pereira AE,
    4. Ramos OL,
    5. Pimentel C,
    6. Gagliardi R,
    7. Sanvito WL,
    8. Lander N,
    9. Mechoulam R

    (1980) Chronic administration of cannabidiol to healthy volunteers and epileptic patients. Pharmacology 21:175–185.

    1. Dennis I,
    2. Whalley BJ,
    3. Stephens GJ

    (2008) Effects of Δ9-tetrahydrocannabivarin on [35S]GTPγS binding in mouse brain cerebellum and piriform cortex membranes. Br J Pharmacol 154:1349–1358.

    1. Egert U,
    2. Heck D,
    3. Aertsen A

    (2002a) Two-dimensional monitoring of spiking networks in acute brain slices. Exp Brain Res 142:268–274.

    1. Egert U,
    2. Knott T,
    3. Schwarz C,
    4. Nawrot M,
    5. Brandt A,
    6. Rotter S,
    7. Diesmann M

    (2002b) MEA-Tools: an open source toolbox for the analysis of multi-electrode data with MATLAB. J Neurosci Methods 117:33–42.

    1. Farrimond JA,
    2. Hill AJ,
    3. Jones NA,
    4. Stephens GJ,
    5. Whalley BJ,
    6. Williams CM

    (2009) A cost-effective high-throughput digital system for observation and acquisition of animal behavioral data. Behav Res Methods 41:446–451.

    1. Feng Z,
    2. Durand DM

    (2005) Decrease in synaptic transmission can reverse the propagation direction of epileptiform activity in hippocampus in vivo. J Neurophysiol 93:1158–1164.

    1. Gordon E,
    2. Devinsky O

    (2001) Alcohol and marijuana: effects on epilepsy and use by patients with epilepsy. Epilepsia 42:1266–1272.

    1. Hampson AJ,
    2. Grimaldi M,
    3. Axelrod J,
    4. Wink D

    (1998) Cannabidiol and (−)Δ9-tetrahydrocannabinol are neuroprotective antioxidants. Proc Natl Acad Sci U S A 95:8268–8273.

    1. Herkenham M,
    2. Lynn AB,
    3. Little MD,
    4. Johnson MR,
    5. Melvin LS,
    6. de Costa BR,
    7. Rice KC

    (1990) Cannabinoid receptor localization in brain. Proc Natl Acad Sci U S A 87:1932–1936.

    1. Hill AJ,
    2. Jones NA,
    3. Williams CM,
    4. Stephens GJ,
    5. Whalley BJ

    (2009) Development of multi-electrode array screening for anticonvulsants in acute rat brain slices. J Neurosci Methods doi:10.1016/j.jneumeth.2009.10.007.

    1. Howlett AC,
    2. Breivogel CS,
    3. Childers SR,
    4. Deadwyler SA,
    5. Hampson RE,
    6. Porrino LJ

    (2004) Cannabinoid physiology and pharmacology: 30 years of progress. Neuropharmacology 47:345–358.

    1. Iuvone T,
    2. Esposito G,
    3. De Filippis D,
    4. Scuderi C,
    5. Steardo L

    (2009) Cannabidiol: a promising drug for neurodegenerative disorders? CNS Neurosci Ther 15:65–75.

    1. Kapur A,
    2. Zhao P,
    3. Sharir H,
    4. Bai Y,
    5. Caron MG,
    6. Barak LS,
    7. Abood ME

    . (2009) Atypical responsiveness of the orphan receptor GPR55 to cannabinoid ligands. J Biol Chem 284:29817–29827.

    1. Karler R,
    2. Cely W,
    3. Turkanis SA

    (1974) Anticonvulsant properties of Δ9-tetrahydrocannabinol and other cannabinoids. Life Sci 15:931–947.

    1. Kwan P,
    2. Brodie MJ

    (2007) Emerging drugs for epilepsy. Expert Opin Emerg Drugs 12:407–422.

    1. Löscher W,
    2. Hönack D,
    3. Fassbender CP,
    4. Nolting B

    (1991) The role of technical, biological and pharmacological factors in the laboratory evaluation of anticonvulsant drugs. III. Pentylenetetrazole seizure models. Epilepsy Res 8:171–189.

    1. Lowry OH,
    2. Rosebrough NJ,
    3. Farr AL,
    4. Randall RJ

    (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275.

    1. Ludányi A,
    2. Eross L,
    3. Czirják S,
    4. Vajda J,
    5. Halász P,
    6. Watanabe M,
    7. Palkovits M,
    8. Maglóczky Z,
    9. Freund TF,
    10. Katona I

    (2008) Downregulation of the CB1 cannabinoid receptor and related molecular elements of the endocannabinoid system in epileptic human hippocampus. J Neurosci 28:2976–2990.

    1. Lutz B

    (2004) On-demand activation of the endocannabinoid system in the control of neuronal excitability and epileptiform seizures. Biochem Pharmacol 68:1691–1698.

    1. Ma YL,
    2. Weston SE,
    3. Whalley BJ,
    4. Stephens GJ

    (2008) The phytocannabinoid Δ9-tetrahydrocannabivarin modulates inhibitory neurotransmission in the cerebellum. Br J Pharmacol 154:204–215.

    1. McCormick DA,
    2. Contreras D

    (2001) On the cellular and network bases of epileptic seizures. Annu Rev Physiol 63:815–846.

    1. Mechoulam R,
    2. Shvo Y

    (1963) Hashish. I. The structure of cannabidiol. Tetrahedron 19:2073–2078.

    1. Monory K,
    2. Massa F,
    3. Egertová M,
    4. Eder M,
    5. Blaudzun H,
    6. Westenbroek R,
    7. Kelsch W,
    8. Jacob W,
    9. Marsch R,
    10. Ekker M,
    11. et al

    . (2006) The endocannabinoid system controls key epileptogenic circuits in the hippocampus. Neuron 51:455–466.

    1. Pertwee RG

    (2008) The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: Δ9-tetrahydrocannabinol, cannabidiol and Δ9-tetrahydrocannabivarin. Br J Pharmacol 153:199–215.

    1. Pohl M,
    2. Mares P

    (1987) Effects of flunarizine on metrazol-induced seizures in developing rats. Epilepsy Res 1:302–305.

    1. Qin N,
    2. Neeper MP,
    3. Liu Y,
    4. Hutchinson TL,
    5. Lubin ML,
    6. Flores CM

    (2008) TRPV2 is activated by cannabidiol and mediates CGRP release in cultured rat dorsal root ganglion neurons. J Neurosci 28:6231–6238.

    1. Rakhshan F,
    2. Day TA,
    3. Blakely RD,
    4. Barker EL

    (2000) Carrier-mediated uptake of the endogenous cannabinoid anandamide in RBL-2H3 cells. J Pharmacol Exp Ther 292:960–967.

    1. Ross HR,
    2. Napier I,
    3. Connor M

    (2008) Inhibition of recombinant human T-type calcium channels by Δ9-tetrahydrocannabinol and cannabidiol. J Biol Chem 283:16124–16134.

    1. Ryan D,
    2. Drysdale AJ,
    3. Lafourcade C,
    4. Pertwee RG,
    5. Platt B

    (2009) Cannabidiol targets mitochondria to regulate intracellular Ca2+ levels. J Neurosci 29:2053–2063.

    1. Ryberg E,
    2. Larsson N,
    3. Sjögren S,
    4. Hjorth S,
    5. Hermansson NO,
    6. Leonova J,
    7. Elebring T,
    8. Nilsson K,
    9. Drmota T,
    10. Greasley PJ

    (2007) The orphan receptor GPR55 is a novel cannabinoid receptor. Br J Pharmacol 152:1092–1101.

    1. Sagredo O,
    2. Ramos JA,
    3. Decio A,
    4. Mechoulam R,
    5. Fernández-Ruiz J

    (2007) Cannabidiol reduced the striatal atrophy caused 3-nitropropionic acid in vivo by mechanisms independent of the activation of cannabinoid, vanilloid TRPV1 and adenosine A2A receptors. Eur J Neurosci 26:843–851.

    1. Scuderi C,
    2. Filippis DD,
    3. Iuvone T,
    4. Blasio A,
    5. Steardo A,
    6. Esposito G

    (2009) Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders. Phytother Res 23:597–602.

    1. Shen M,
    2. Thayer SA

    (1999) Δ9-Tetrahydrocannabinol acts as a partial agonist to modulate glutamatergic synaptic transmission between rat hippocampal neurons in culture. Mol Pharmacol 55:8–13.

    1. Smith I,
    2. Bevan SA,
    3. Whalley BJ,
    4. Stephens GJ

    (2009) Phytocannabinoid affinities at CB1 receptors in the mouse cerebellum. Proceedings of the 19th Annual Meeting of the International Cannabinoid Research Society; 2009 July 8–11; Pheasant Run, St. Charles, IL. P81, International Cannabinoid Research Society.

    1. Stett A,
    2. Egert U,
    3. Guenther E,
    4. Hofmann F,
    5. Meyer T,
    6. Nisch W,
    7. Haemmerle H

    (2003) Biological application of microelectrode arrays in drug discovery and basic research. Anal Bioanal Chem 377:486–495.

    1. Straiker A,
    2. Mackie K

    (2005) Depolarization-induced suppression of excitation in murine autaptic hippocampal neurones. J Physiol 569:501–517.

    1. Thomas A,
    2. Baillie G,
    3. Philips AM,
    4. Razdan RK,
    5. Ross RA,
    6. Pertwee RG

    (2007) Cannabidiol displays unexpectedly high potency as an antagonist of CB1 and CB2 receptor agonists in vitro. Br J Pharmacol 150:917–926.

    1. Tsou K,
    2. Brown S,
    3. Sañudo-Peña MC,
    4. Mackie K,
    5. Walker JM

    (1998) Immunohistochemical distribution of cannabinoid CB1 receptors in the rat central nervous system. Neuroscience 83:393–411.

    1. Vivo M,
    2. Lin H,
    3. Strange PG

    (2006) Investigation of cooperativity in the binding of ligands to the D2 dopamine receptor. Mol Pharmacol 69:226–235.

    1. Wallace MJ,
    2. Martin BR,
    3. DeLorenzo RJ

    (2002) Evidence for a physiological role of endocannabinoids in the modulation of seizure threshold and severity. Eur J Pharmacol 452:295–301.

    1. Wallace MJ,
    2. Wiley JL,
    3. Martin BR,
    4. DeLorenzo RJ

    (2001) Assessment of the role of CB1 receptors in cannabinoid anticonvulsant effects. Eur J Pharmacol 428:51–57.

    1. Watanabe K,
    2. Kayano Y,
    3. Matsunaga T,
    4. Yamamoto I,
    5. Yoshimura H

    (1996) Inhibition of anandamide amidase activity in mouse brain microsomes by cannabinoids. Biol Pharm Bull 19:1109–1111.

    1. Whalley BJ,
    2. Stephens GJ,
    3. Constanti A

    (2009) Investigation of the effects of the novel anticonvulsant compound carisbamate (RWJ-333369) on rat piriform cortical neurones in vitro. Br J Pharmacol 156:994–1008.

Cannabindiol

  • August 24, 2012 3:41 am

CBD (cannabidiol) – Cannabidiol is the cannabinoid obtaining most recent mainstream exposure for its multitude of benefits with non-psychoactive effects. Unites States Patent #6,630,507 – Hampson, et al. found cannabinoids (specifically CBD) effective as antioxidants & neuroprotectants without providing any psychoactive effects. Neuroprotective antioxidant more potent than ascorbate or topopherol with ability to pass the blood-brain barrier. Works as a CB1 antagonist in the presence of THC at low concentration (nM). Modulates THC adverse events including anxiety, tachycardia, hunger & sedation.  CBD increases endogenous cannabinoid levels by two mechanisms: as a weak uptake inhibitor of AEA and an inhibitor of its hydrolysis.  Acts as a TRPV1 agonist (similar to capsaicin, lacking noxious effects).

Similar Readily Cannabinoids Penetrate into the Brain

  • August 24, 2012 3:31 am

The pharmacokinetics (behaviour of a substance in the body) of cannabidiol (CBD), cannabidivarine (CBDV), delta-9-
tetrahydrocannabivarin (delta-9-THCV) and cannabigerol (CBG) in mice and rats were investigated at the University of Aberdeen, UK.
Researchers determined concentrations in the brain after intraperitoneal (injection into the abdomen) and oral administration.
The effects of CBD were further investigated in an animal model of obsessive compulsive behaviour.

All phytocannabinoids readily penetrated the blood-brain barrier. In rats, oral administration offered higher brain concentrations for CBD
and CBDV, but not for delta-9-THCV and CBG, for which the intraperitoneal route was more effective. CBD inhibited obsessive-
compulsive behaviour in a time-dependent manner matching its concentration in the brain.

(Source: Deiana S, Watanabe A, Yamasaki Y, Amada N, Arthur M, Fleming
S, Woodcock H, Dorward P, Pigliacampo B, Close S, Platt B, Riedel G.
Plasma and brain pharmacokinetic profile of cannabidiol (CBD),
cannabidivarine (CBDV), (9)-tetrahydrocannabivarin (THCV) and
cannabigerol (CBG) in rats and mice following oral and
intraperitoneal administration and CBD action on obsessive-compulsive
behaviour. Psychopharmacology (Berl). 2011 Jul 28. [in press])

Therapeutical Effects of the Main Cannabinoids:

 

CBD
(Cannabidiol):
 Non-psychoactive. Reduces muscle spasms. Muscle relaxant. Analgesic (pain relieving) Trends in Pharmacological Sciences
Volume 30, Issue 10, October 2009, Pages 515-527

CBN
(Cannabinol): Mildly psychoactive. Non-narcotic analgesic (pain relieving). Good indication of medications age.
 Trends in Pharmacological Sciences
Volume 30, Issue 10, October 2009, Pages 515-527

THC
(tetrahydrocannabinol): Psychotropic. Analgesic (pain relieving). Apatite stimulant. Bronchial dilator. Lowers IOP/glaucoma.

THCV
(Tetrahydrocannabivarin): Stronger, faster “high
” effect.Apatite suppressantEuphoriaanalgesic (pain relieving).

CBG
(Cannabigerol): Non-psychoactive, sleep inducing. Anti-microbial. Lowers intra-ocular pressure (IOP) Glaucoma. Trends in Pharmacological Sciences Volume 30, Issue 10, October 2009, Pages 515-527

CBC (Cannabichromene): Sedative effectModerates effects of THC.Analgesic (pain relieving). Non-psychoactive. Trends in Pharmacological Sciences
Volume 30, Issue 10, October 2009, Pages 515-527

 

Medical applications of the non-psychotropic cannabinoids CBD, THCV, CBC, CBG, CBDV and THCA.

Cannabindiol Crosses the Blood Brain Barrier

  • August 24, 2012 3:27 am

11/08/08
Dr. Courtney speaks on the CANNABIS ISSUES PANEL of experts and authors at the 18th ANNUAL SOUTHERN HUMBOLDT HEMP FEST

Emcee: Jimmy Durchslag:

Next we have William Courtney, DR. BILL COURTNEY, Cannabis Medical Consultant. Many of you… I know he’s based in Mendocino, has an office in Willits, but he’s also here in our area, regularly. He’s my medical consultant. And I know if you’ve e’er seen him, you get a lot of information about cannabis–it’s health effects. So he’s going to talk to us about that. Dr. Bill. Welcome. (applause)

Dr. Bill:

Thank you. I just noticed today that this is the 18th Annual Hemp Fest. Interestingly, the International Cannabinoid Research Society has also had it’s 18th Annual.

And this is the manual that came out of the 18th Annual research [He shows the bound sheaf of papers.] And it’s free online. If you go to [sounds like "Icarus"] ICRS, and go to the symposium page, like you were going to sign up for the symposium, this is a PDF.

And for those who are not able to go online, there are a stack of cd’s over there. You can take this to Staples and for $20 print your own copy.

“Icarus” [ICRS], this year, was phenomenal! It was only overshadowed by my discovery of the United States Patent on CBD and cannabinoids.

But, since there is so much to talk about, I want to focus a little bit on Icarus, first, and then we’ll get into the U. S. Patent.

The thing that came out of this years’ ICRS that was quite amazing is there’s a whole series of acid molecules in the raw plant – the 21 carbon molecules. Those break down with the slightest amount of heat or aging. THC-A, which previously was thought of as a storage molecule, is in fact and active molecule, and when you heat it, you break that carboxy group off to create THC, which is psychoactive. So a lot of our activities over the last 40 years have been aimed at how to improve the thc potency, because we had assumed that the psychoactivity was tantamount to the medicinal aspects.

But as we look at it closer it turns out that there’s an awful lot about the green plant, before it’s been treated culturally, with heat, that has a lot of benefit. And this years’ Icarus introduced a lot of these: cannabidiol acid, cannabichromic acid, cannabigerol acid: all of those 21 carbon molecules have unique medical properties. In particular, CBD-A, or cannabidiol acid, is an antibiotic. As soon as you put it into a tea, you simmer it, you saute it, you bake it, you smoke it, you lose that antibiotic function, and it converts it into CBD; with additional heat it converts into thc; and so with prolonged heating you increase the amount of THC (but our plant currently has a lot of THC, so that really isn’t essential.

My current position, for those of you who I’ve met with clinically, I’m recommending that the greens be part of your diet.

If you look at the world around us you see that the deer eat it; they don’t really eat a lot, but they kind of munch at it. Iguanas eat it. Birds eat it. Rats. Dogs. Cats. Animals understand that.

A few of the leaves allows you access to, not only THC, but the THC-A, these other cannabinoids, all of which are fat molecules, that move into the adipose tissue of your body. And if you eat it regularly for two months, you eventually will become saturated, which is a desirable state in which these various chemicals can be found, and are plentiful, in the fat tissue of the body.

The benefit of them being fat molecules that store in the fat is that it acts like a depo, in the sense that if your body needs a particular supplement, it can draw down from the fat tissue that particular cannabinoid. And these cannabinoids, we now know, provide anti-epileptic, they provide antibiotic, antiviral, antifungal, anti-anxiolitic, anti-psychotic, a tremendous range of medicinal activities.

And there was an article out of last years’ Icarus that talked about the unique ability of body to draw down the specific cannabinoid as it’s needed. So, ideally, if it’s a part of your diet, the way that it is with the rest of the animals who are not enculturated, if you have a minor fungal infection, you’ll pull down an antifungal before it becomes clinical, and you’ll prevent the problem from becoming even noticeable. The same with an anti-psychotic, the same with an anti-anxiolitic (anti-anxiolitic is something that reduces anxiety, cuts anxiety. So, it’s such a wide range of conditions that this plant is capable of contributing to our body’s ability to modulate.

And I think that’s one of the take-home messages of this last Icarus, is that all this plant does is facilitate the body’s ability to take care of itself.

If each person here would take ten hours and review the immune system on the web, go to Wikepedia, look up all the links, learn about the immune system, how anti-inflammatory, auto-immune, how it detects self, not-self, precancerous changes. I mean, the immune system is absolutely amazing. And what cannabis does is facilitate the regulation of the immune system.

The body produces endogenous cannabinoids. You may have heard of endorphins: that’s endogenous morphine. Well, in a similar fashion, the body produces endogenous cannabinoid.

I’m currently submitting an article for publication which would contract that to “encannans”, and the reason that I selected that word is because it removes part of this tone of cannabis, which causes such panic in so many people, and, in order to reach out across the aisle to the Republicans. . . You know you want to not insult them too strongly from the get-go, that first impression is hard to redo. . .

But also part of that article is the history of these molecules. Lipins are the little fat messenger molecules. Those are found in archeia, which is a primitive single-celled organism that was in existence four billion years ago. These little fat molecules would attach to a protein receptor, which we now call a cannabinoid receptor, and the structure of those (they go back and forth through the membrane seven times) that specific structure is still found in the proteins in the membrane of your T cells and D cells (white blood systems in your immune system found in your lymph nodes, spleen, thiamus).

So the entire immunologic system in the human being communicates through the CB2 receptor, which is similar to the same receptor that was first evolved (if evolution is a part of your world view) billions and billions of years ago. So we had this incredibly conserved system (and conservation is when a design is so effective that it doesn’t change over time). Most biological designs evolve over maybe a couple million years, and they turn over and evolve, but this was so effective and so incredible that it has. . . 4 billion years later you can find it in the cells that are involved in fighting cancer, and that are fighting infection.

I may be getting a little bit detailed here, but all to say, if you want to look at this Icarus, and I highly, highly recommend that you do this, send me an email, and I can send you this as a pdf file that you can just look at electronically, or you can take it to Staples, print it out, and for those of us who like to look at a physical copy, you can make notes and go back and forth.

We’re hoping to bring the Second International CB2 Conference to Mendocino. I attended the First International, which was in Banff in May of ’06, and Ive been speaking with Keith Sharkey, who coordinated that. Researchers from all over the world, from Israel, Japan, Germany, United Kingdom, Canada, all convened for three days and we did nothing but exchange information about this 2nd cannabinoid binding receptorthat CB2 receptor we were just talking about–thats in the immune system.

Because medicine is nothing but about enhancing immunology, and immunology is nothing but about detecting self/non-self, thats foreign bodies, protection from cancers, and trying to make a peace in the world where your own cells can decide to defy.

A million times a day, cells begin to divide that shouldn’t be dividing and the immune system has to say, “Hey, you know, the shrubs are getting a little hairy there; we’ve got to trim them down. We’ve got enough renal cells or kidney cells. We’ve got enough bone cells, muscle cells.” So the immune system’s highly competent at dealing with that, but it can become more competent. And that is exactly what cannabinoids do, whether you’re talking about the body’s endogenous cannabinoids, or the plant, which can be phytocannabinoids (which just means cannabinoids from plants) or exogenous cannabinoids. Both of those terms refer to those 20 carbon molecules that are produced outside of the body, but bind to the protein receptors on the cells in our body and therefore help regulate it. So whether we’re talking about the body’s chemistry, or the plant chemistry, their role is to modulate the function of the immune system. And a modulator is a chemical substance that can either increase the activity of the immune system, or decrease it.

And it’s like, medicine always bothered me when I would study traditional pharmaceuticals, and they’d say, “Well, they can give diarrhea or constipation.” Well, that’s great, you know, huh!? Can you choose, or do you just kind of get stuck with what you get stuck with? Well, the same with. . . A modulator is a substance which provides feedback. Every cell in the body has a function: brain cells, nerve cells, blood cells, bone cells, muscle cells. There’s something that they’re supposed to be doing. And it could be overactive. And, if it’s overacting, the body would like to tone it down, slow it down. And the feedback it gets from the cannabinoids say, “Wow, this is too much, we need to slow it down, dial it in.” If the system is insufficiently active, then it wants to speed it up to normal.

So a cannabinoid is a modulator that restores optimal function. That’s what the body’s system is doing. That’s what the plant does. The plant facilitates the rapid restoration of the normal function.

And I focus a lot on the immune system because that’s so central to disease and illness, but it modulates the endocrine system. It modulates the neuromuscular system. It modulates bone remodeling – osteoporosis and osteomalacia. Those are conditions in which there are two cells, one of which burrows through the bone and chews up the old bone, then a new one, the osteoblast comes and lays down new bone, and so the bone is completely rebuilt every seven years. That’s the bone life-cycle, the longest in the body. And these two cells are involved in that constant remodeling. Well, they communicate by cannabinoids. It’s become the “duh!” [laughs] of modern physiology.

Everything uses these very simple fat molecules to provide feedback, and as we age with the loss of some of the female hormones, in particular, that communication falters, and the osteoclasts (that cell that burrows and dissolves the bone) just keeps tunneling away, and the osteoblast just kind of kicks back and gets out of sync and doesn’t lay down new bone, and we perforate, making the bone porous. And so the cannabinoids facilitate that one-to-one communication, restore, (Restore may be a leap. Lots of times I get overzealous in my hopes.) at least will facilitate the communication which will prevent further porosity changes. It will stop the condition from progressing and, if the design is consistent, I actually believe it will increase the density and restore normal density with that communication. Because, if you’re burrowing, and you cross a tunnel, you don’t have to burrow through that tunnel because it’s missing, but the new one will come through and lay down new bone.

So, I’m going to stick with my thing. I think it will actually improve the condition. So, bone-remodeling, cancers, inflammatory conditions, prostate cancer, growth regulation, metastasis, all that, can be found in this free book on the web. Consider this the most gracious gift of the web to people who have an interest in health. And, like I said, just send me your email, I’ll send you an electronic copy that you can peruse.

And, we hopefully will review this before the 2nd Cannabinoid Conference. I’ll try to do a one to two or three day review of the language, and we’ll go from the terms, the definitions; we’ll look at a little bit of chemistry, then we’ll look at a little bit of microbiology (which is a big love of mine) and how biochemistry is involved in the functions of the cell: all that to kind of prepare us for this group of visiting dignitaries which, you know, will tell us where the dream is today.

Something that is incredibly overwhelming is the discovery of Patent 6,630,507B1. The United States of America has been assigned the patent rights to cannabinoids as antioxidants and neuroprotectants.

Over on the table there is a face sheet, these are free hand-outs. You go home. You type in USPTO (which is US Patent and Trademark Office) and google, it will take you to their site. Search by patent number. Just type that patent number in. It’ll bring you up the electronic copy of this patent (for those of you can do that), for those who can’t, there are hard copies over there and if you want to make a donation for print and paper you can. But these are primarily for people who don’t have access to the web, and are not into electronic copies. This should be read probably a minimum of ten times, and then committed to memory. It really is absolutely phenomenal. It reminded me of more organic chemistry than I ever knew. Whenever you write a patent, you hire a very expensive attorney, who really is an English literature graduate doctoral person, and there are some patent attorneys who aren’t like that, but in general their skill is in being very articulate.

And this talks about the chemistry of, particularly, cannabidiol. On two separate occasions the government lists cannabidiol on separate line item claims. It spells out the conditions for which cbd is uniquely beneficial – the oxidated stress diseases–and that includes everything from diabetes, rheumatoid arthritis, the neo-plasias, coronary vascular disease.

In particular they isolate the neuro conditions.

CBD can cross the blood-brain barrier. So, it’s a fat molecule that can get into the brain, which is very hard to do, because the brain is very protective, but because this has 34 million years of research on its side it knows how to walk between the lines and the body has allowed it access to the brain. So, for Parkinsons, for Alzheimers, for strokes whether embowelic, hemmorhagic, or traumatic, or surgical, these are all conditions where CBD is uniquely beneficial.

For the first time ever in three years of looking, this has human dosage schedules. This is of phenomenal importance to you legally. It tells out what you are entitled to do, and then you just look at the plant, and the plant tells you how much plant material you need to generate that.

Speeding quickly forward through many other things, part of what I talk to all my patients about is the ability to fractionate for cbd, which means. . . There’s a piece of paper over there which tells you the technique. That’s another thing to read and memorize, because it’s the single most important thing for your health, and for your legal safety.

You take fine grind cannabis.

At my office there’s this $400 thermometer, which is supposed to be very accurate. You get a variable temperature heat gun, which allows you to provide a constant single temperature for hours at a time, so that it’s better than any of the vaporizers, because it’s very, very, very steady. So you come in here. You turn the heat on until it goes to 166 degrees Centigrade. Take the file; mark that place on your heat gun, so you can get back to that.

Take your cannabis. You grind the cannabis to a fine degree which increases the surface area so that you can get to the cannabinoids in the plant material.

Put the heat to it. The longer you heat it the more THC is boiled.

We’ve now gotten down to the physics of the cannabinoids. The vaporization temperature is a temperature at which a liquid goes through a phase change to a gas. Like with water, at a 100 degrees Centigrade water begins to boil. You’ll never get water at 101 degrees because it turns into a vapor before it can get that hot.

THC boils at 157 degrees, that’s Delta 9. Delta 8 boils at a 175. Cannabidiol at 188. So if you have a very specific temperature, you can heat the cannabis and vaporize the THC. If you want THC, you go ahead and use that, or do whatever you can do with that, but there’s so much of it there that for most people, we need to lower the THC content so you can get to the CBD.

So once you’ve heated it you’ve stripped the THC out of it. You now have cannabidiol plant material. That CBD plant material is nonpsychoactive, and it’ll allow you to begin to get to the dosage schedule that this patent recommends.

For the last year I’ve been recommending 5 milligrams per kilogram body weight. That is an amount of cannabidiol that it takes for this particular mouse strain to block the development of diabetes.

Now, I’ve been hanging my hat on that for a year, but I now have 5,000 pegs to hang my hat on, thanks to the United States government patent. Because. . . They talk about higher doses. They talk about 10 milligrams per kilogram body weight. They talk about 20 and 40. Huge. Hugely important!

But if you just stick with the 5 mg/kg, that still requires you to process a lot of plant material, because the amount of cbd is so low. There is a strain called “White Russian”, which is 100% more CBD, and there’s some new strains coming out of Mendocino which could have up to 700% more CBD. These strains will make it a lot easier to get access to the 500 mg a day, which is the schedule that I’ve been recommending to my patients.

That’s the same amount that you would use of vitamin C, and at that level, in this particular mouse strain, you block the development of diabetes 60% of the time. There’s a particular mouse strain that 86% of these mice develop diabetes. The pharmaceutical industry tests all oral and injectable medications on this mouse strain because it’s such a reliable model for diabetes, so, you know, does glucophage inhibit diabetes? Does insulin/hemulin? It’s an amazing animal model for diabetes.

If you give that animal model which 86% of the time will develop diabetes, if you give them 5 mg/kg body weight, 60% of them do not go on to develop diabetes. It completely blocks the. . .[audience applause]

Yeah, 14 million Americans go blind with diabetes a year! Kidney renal failure. I’ve got patients I see in their homes who are on dialysis, people who’s knees have been cut off, people who’s vision fails because of diabetic retinopathy.

And, we could prevent that with something that’s non psychoactive, and that’s just an amazing molecule, as is thoroughly detailed in this patent. Which is why every person here needs to just know this thing inside and out. And it will take you a while. I mean, I’m still studying it and every time I read it there’s more stuff there. It really is phenomenal.

I have a room upstairs at 4:20, and any specific questions on either fractionating for cannabinoids (and pick up that sheet and study that–key to your legal and central to your physical health). . . There’s one over there on green leaf.

Did we talk about green leaf yet?

So if you talk about those molecules that are present in the fresh green leaf, I try to use 8 leaves a day. I juice it with carrots. I started out with carrots – I do four in the morning, four in the afternoon, but after eating carrot juice for five or six months you begin to turn orange a little bit (I knew someone in college who actually did), and so I now put in beets and broccoli (just because Bush doesn’t like it, it figures it’s got to be good for you), anything laying around – kales. It makes a drink.

And it’s phenomenal because you get these cannabinoids in, they’re non-psychoactive, they saturate the body and they facilitate the body’s regulation. And so, I tell each of my patients: the single most important thing you put in your mouth after water, on a daily basis, is, approximately, eight green leaves a day.

That number is unknown, but I’ve had 90 year old people who have gotten out of their wheelchairs who’ve had arthritis that was so inflammatory and painful. . . By down-regulating the inflammatory system you decrease the swelling and inflammation, you increase mobility and articulate function. Its prevention of alzheimers, of coronary vascular disease. Prophylactic uses.

Prophylactic uses are the most pivotal in this document. I don’t know how they’ll stand up in court but I plan on using them very soon, like November 6th Im going to be in court presenting this information on this prophylactic use. That’s until they take my license away. But then I’ll keep doing it, so…

If you have any questions, talk to me later upstairs, and I’ll answer them if I’m able. [audience applause]

emcee Jimmy:

Dr. Courtney! Thank you, Bill. Obviously a wealth of information there. He has his very attractive banner up here. Is that drcourtney@mcn.org? yes, if you want a copy of the Icarus document which he has offered on line. His card is there. There’re a couple books here that he’s involved with, and he’ll be upstairs after this panel to answer questions and give more information.

The workshop room is up in that corner. 4:20, he said. I don’t know how that time got chosen, probably just randomly. So, thank you for that.

 

 

 

Source:http://www.civilliberties.org/courtney.html

The Human Endocannabinoid System

  • June 18, 2012 5:58 am

The Endocannabinoid System (ECS) started revealing itself to researchers in the 1940s and by the late ‘60s the basic structure and functionality had been laid out. Today we know the ECS is a comprehensive system of biochemical modulators that maintain homeostasis in all body systems including the central and peripheral nervous systems, all organ systems, somatic tissues, and all metabolic biochemical systems, including the immune system.

This homeostatic matrix is not a recent evolutionary twist just for humans; we find the Endocannabinoid System in every chordate creature for the last 500 million years. It is a fully mature biochemical technology that has maintained health and metabolic balance for most of the history of life itself.

The two major interactive systems within the ECS are (1) the cannabinoid receptors that we find on all cell surfaces and neurological junctions and (2) the endocannabinoids that fit the receptors to trigger various metabolic processes. Looking at a cannabinoid receptor distribution map we see that CB1 receptors, that are most sensitive to anandamide, are found in the brain, spinal nerves, and peripheral nerves. CB2 receptors preferred by 2-arachidonoylglycerol (2-AG) are found largely in the immune system, primarily the spleen. A mix of CB1 and CB2 receptors are found throughout the rest of the body including the skeletal system. And yes, 2-AG or CBD will grow new trabecular bone.1 It is also useful to note that both anandamide and 2-AG can activate either CB1 or CB2 receptors.

The nature of the endocannabinoids are functionally much like neurotransmitters, but structurally are eicosanoids in the family of signaling sphingolipids. These signaling cannabinoids keep track of metabolic systems all over the body. This information is shared with the nervous system and the immune system so that any imbalance is attended to. If the body is in chronic disease or emotional stress, the immune system can fall behind and lose control of compromised systems. It is here that phytocannabinoids can pitch in to support the stressed body in a return to health. The cannabis plant provides analogues of the body’s primary signaling cannabinoids. Tetrahydrocannabinol (THC) is mimetic to anandamide, and cannabidiol (CBD) is mimetic to 2-AG, and has the same affinity to CB1 and CB2 receptors; providing the body with additional support for the immune and endocannabinoid systems.

Phytocannabinoids supercharge the body’s own Endocannabinoid System by amping up the response to demand from the immune signaling system in two modes of intervention: one, of course, is in bonding with the cannabinoid receptors; the other is in regulation of innumerable physiological processes, such as cannabinoid’s powerful neuroprotective and anti-inflammatory actions, quite apart from the receptor system. It is interesting to note here that the phytocannabinoids and related endocannabinoids are functionally similar, but structurally different. As noted above, anandamide and 2-AG are eicosanoids while THC and CBD are tricyclic terpenes.

Let us look more closely at the two primary therapeutic cannabinoids, THC and CBD. The National Institutes of Health tell us that THC is the best known because of its signature psychotropic effect. This government report shows THC to be effective as an anti-cancer treatment, an appetite stimulant, analgesic, antiemetic, anxiolytic, and sedative.2

CBD (cannabidiol) is a metabolic sibling of THC, in that they are alike in many ways but are also different in important properties. First we see that CBD has no psychotropic effects and there are few CB2 receptors in the brain and peripheral nerves. There appears to be a broader therapeutic profile associated with CBD, which is listed here:3

 

anxiolytic

antipsychotic

antiepileptic

neuroprotective

vasorelaxant

antispasmodic

anti-ishcemic

antiproliferative

antiemetic antibacterial

anticancer

antidiabetic

antisporiatic

intestinal anti-prokinetic

analgesic

bone-stimulant

anti-inflammatory

immunosupressive

 

One of the most important health benefits of cannabinoids is their anti-inflammatory property. In this, they are strong modulators of the inflammatory cytokine cascade. Numerous disease states arise out of chronic inflammation; such as, depression, dementias including Alzheimer’s, cancer, arthritis and other autoimmune disorders, viral infection, HIV, brain injury, etc.

Inflammatory cytokines can be activated by oxidative stress and disease states. Cannabinoids, being immunomodulators interrupt the cytokine inflammatory cascade so that local inflammation does not result in tissue pathology. Thus we are spared morbid or terminal illnesses.4

If our own endocannabinoid system can maintain metabolic homeostasis and even cure serious disease, why are we plagued by illness? We know that the body produces only small amounts of anandamide and 2-AG; enough to maintain the body but not enough to overcome chronic stress, illness, injury, or malnutrition. Cannabls is the only plant we know of that produces phytocannabinoids that mimic our own endocannabinoids. One of the great benefits of this mimetic medicine is that cannabinoids are essentially natural to our biology and do no harm to our tissues and systems.

It is well known that most diseases of aging are inflammatory in origin, thus making cannabis the best anti-aging supplement we could take to avoid arthritis, dementia, hypertension, diabetes, osteoporosis, and cancer. This is our key to good health and long life.

Since it is such an important attribute, as well as being independent of the cannabinoid receptor system, let’s look a little deeper into the ability of cannabinoids to inhibit the inflammatory cytokine cascade. Inflammation is good for us, a little here, a little there; it brings T-cells and macrophages to infection sites. This is good. However, chronic inflammation can cause serious illness and death. How do phytocannabinoids rescue us from dreaded infirmities? When the call comes in to the immune system to send troops, the first thing to happen is that the immune system signals glial cells to produce cytokines. Once this cat is out of the bag, the process can go one of two ways.

A) Killer cells clean up the infection and all is well.

B) Cytokines can stimulate more cytokine production and cause many more cytokine receptors to awaken. Unchecked, this becomes a cytokine storm showing symptoms of swelling, redness, fatigue, and nausea; even death.

Phytocannabinoids have the ability to suppress this inflammatory cytokine cascade by inhibiting glial cell production of the cytokines interferon or interleukin. Here we see the seeds of chronic inflammation dissolved by the modulation process of cannabinoids bringing homeostasis to systems out of balance. This is a good example of how cannabinoids normalize biological processes all throughout the body and allows us to keep that glow of well-being through a long and happy lifetime.

~Dennis Hill

References:

1. http://www.pnas.org/content/103/3/696

2. http://www.cancer.gov/cancertopics/pdq/ … onal/page1

3. http://cannabisinternational.org/info/N … inoids.pdf

4. http://www.ncbi.nlm.nih.gov/pubmed/16918439

THC and Cannabinol Activate Capsaicin-Sensitive Sensory Nerves via a CB1 and CB2 Cannabinoid Receptor-Independent Mechanism

  • June 18, 2012 5:46 am
  1. Peter M. Zygmunt,
  2. David A. Andersson, and
  3. Edward D. Högestätt

+ Author Affiliations


  1. 1 Department of Clinical Pharmacology, Institute of Laboratory Medicine, Lund University Hospital, SE-221 85 Lund, Sweden
 Abstract

Although Δ9-tetrahydrocannabinol (THC) produces analgesia, its effects on nociceptive primary afferents are unknown. These neurons participate not only in pain signaling but also in the local response to tissue injury. Here, we show that THC and cannabinol induce a CB1/CB2 cannabinoid receptor-independent release of calcitonin gene-related peptide from capsaicin-sensitive perivascular sensory nerves. Other psychotropic cannabinoids cannot mimic this action. The vanilloid receptor antagonist ruthenium red abolishes the responses to THC and cannabinol. However, the effect of THC on sensory nerves is intact in vanilloid receptor subtype 1 gene knock-out mice. The THC response depends on extracellular calcium but does not involve known voltage-operated calcium channels, glutamate receptors, or protein kinases A and C. These results may indicate the presence of a novel cannabinoid receptor/ion channel in the pain pathway.

  • calcitonin gene-related peptide
  • cannabinoids
  • cannabinol
  • cannabis
  • capsaicin
  • nociceptors
  • pain
  • receptors, sensory
  • tetrahydrocannabinol
Previous SectionNext Section

Introduction

Marijuana contains a mixture of different cannabinoids, of which Δ9-tetrahydrocannabinol (THC), the major psychoactive ingredient, has been characterized extensively with regard to analgesic and anti-inflammatory effects (Mechoulam and Hanus, 2000;Pertwee, 2001). The presence of CB1cannabinoid receptors in the pain pathway may explain the analgesic effects of cannabinoids (Zimmer et al., 1999; Morisset et al., 2001;Pertwee, 2001). However, the well known psychotropic effects of many cannabinoids are attributable to activation of CB1 receptors and limit their therapeutic value as analgesics (Pertwee, 2001). Interestingly, some cannabinoids, such as cannabidiol, cannabinol, and carboxy derivatives of THC, have analgesic and anti-inflammatory effects despite being weak CB1 receptor agonists (Srivastava et al., 1998;Burstein, 1999; Malfait et al., 2000). The effect of THC in the hot-plate test is lost in CB1 receptor gene knock-out mice (Ledent et al., 1999; Zimmer et al., 1999), but the analgesic effect of THC in the tail-flick test is intact (Zimmer et al., 1999). This indicates that THC can induce antinociception also via a CB1 receptor-independent mechanism.

Although CB1 receptors are present on a subpopulation of primary sensory neurons, the effects of THC on pain-sensing primary afferents have not been examined. In addition to transmitting nociceptive information to the CNS, these nerves also participate in the local response to tissue injury, including the release of vasodilator neuropeptides (Holzer, 1992; Szallasi and Blumberg, 1999). Thus, primary sensory nerves are able to release neuropeptides, such as calcitonin gene-related peptide (CGRP) and substance P, in both the periphery and the spinal cord (Holzer, 1992;Szallasi and Blumberg, 1999). In the vasculature, this leads to vasodilatation and increased vascular permeability (Holzer, 1992). Isolated arterial segments provide a sensitive bioassay for studying the effects of drugs acting on such efferent signaling (Hogestatt and Zygmunt, 2002). Initially, using this bioassay, we planned to study whether cannabinoids, including THC and HU-210, inhibit the activity of perivascular sensory nerve. Unexpectedly, we found that THC itself causes activation of capsaicin-sensitive sensory nerves. This effect of THC is not mediated by known cannabinoid receptors and could indicate the existence of a novel target for cannabinoids in the pain pathway.

MATERIALS AND METHODS

Animals. Experiments were performed on hepatic and mesenteric arteries from female Wistar–Hannover rats (250 gm) obtained from M & B (Ry, Denmark) and on mesenteric arteries from male mice (30 gm). Wild-type mice (C57BL/6J) were obtained from M & B, whereas Professor David Julius (University of San Francisco, San Francisco, CA) generously supplied vanilloid receptor subtype 1 gene knock-out (VR1−/−) mice and their homozygous controls (VR1+/+). The genotype (VR1−/− or VR1+/+) was not disclosed until the experiments had been completed.

Recording of tension. The arteries were cut into ring segments and mounted in tissue baths containing physiological salt solution (PSS) of the following composition (in mm): NaCl 119, NaHCO3 15, KCl 4.6, NaH2PO4 1.2, MgCl2 1.2, CaCl2 1.5, and (+)-glucose 6.0. The PSS was continuously bubbled with a mixture of 95% O2 and 5% CO2, resulting in a pH of 7.4. All experiments were performed at 37°C in the presence ofN G-nitro-l-arginine (300 μm) and indomethacin (10 μm) to eliminate any contribution of nitric oxide and cyclooxygenase products, respectively. Relaxations were studied in vessels contracted with phenylephrine (3 μm). When stable contractions were obtained, agonists were added cumulatively to determine concentration–response relationships. Unless otherwise stated, the effects of test substances on vasorelaxation were recorded after pre-exposure of the vessels to the test substances or vehicle for 30 min. Each vessel segment was exposed to only one treatment. In some experiments, the endothelium was removed by blowing carbogen through the vessel lumen. Lack of relaxation in response to 10 μm acetylcholine confirmed a successful removal of the endothelium.

Measurement of CGRP. Segments of rat hepatic or mesenteric arteries were equilibrated for 1 hr in aerated PSS (95% O2 and 5% CO2; 37°C; pH 7.4) containingN G-nitro-l-arginine (300 μm) and indomethacin (10 μm). After a 20 min preincubation period with test drugs in PSS, preparations were transferred to Eppendorff tubes containing the test drugs or vehicle and 0.05% bovine serum albumin in either PSS, nominally calcium-free PSS (10 μmEGTA), or Tris-buffer solution (experiments with lanthanum). The segments were removed after 10 min, and the solution in the test tubes was evaporated. The amount of CGRP in the pellet was determined using a rat 125I-labeled CGRP radioimmunoassay kit (Peninsula Laboratories, Belmont, CA). The Tris-buffer solution was of the following composition (in mm): NaCl 134, Trisma base 5 mm, KCl 4.6, MgCl2 1.2, CaCl2 1.5, and (+)-glucose 6.0, pH 7.4.

Calculations and statistics. Relaxations are expressed as percentage reversal of the phenylephrine-induced contraction. The maximal relaxation (E max) and the log molar concentration of drug that elicited half-maximal relaxation (pEC50) were calculated using GraphPad Prism (version 3.00; GraphPad Software Inc., San Diego, CA). When the concentration–response curve did not reach a plateau, and henceE max and pEC50could not be determined, the area under the curve was calculated (GraphPad Prism version 3.00) and used for evaluation of drug effects. Data are presented as mean ± SEM (vertical lines in figures), andn indicates the number of experiments performed (number of animals). Statistical analysis was performed using Student’s unpairedt test (two-tailed) or ANOVA followed by Bonferroni’s test (GraphPad Prism version 3.00). Statistical significance was accepted when p < 0.05.

Drugs. Phorbol 12,13-dibutyrate (PDBu), 4α-phorbol 12,13-dibutyrate, and staurosporine (Biomol, Plymouth Meeting, PA); anandamide (Cayman Chemical, Ann Arbor, MI); SR141716A (Sanofi Winthrop, Montpellier, France); cannabinol (−)-Δ9-tetrahydrocannabinol, 11-OH-Δ9-tetrahydrocannabinol, and Δ9-tetrahydrocannabinol-11-oic acid (Sigma, St. Louis, MO); and capsaicin, capsazepine, ryanodine, and AM251 (Tocris, Bristol, UK) were all dissolved in and diluted with ethanol. Distilled water or saline was used as solvent for α-latrotoxin, calcicludine, ω-conotoxin GVIA, and ω-conotoxin MVIIC (Alomone Labs, Jerusalem, Israel); nimodipine (Nimotop; Bayer, Wuppertal, Germany); indomethacin (Confortid; Dumex, Copenhagen, Denmark);l-phenylephrine hydrochloride, acetylcholine hydrochloride,N G-nitro-l-arginine, caffeine, rat CGRP, human 8-37 CGRP, l-glutamic acid, and ruthenium red (Sigma); CNQX disodium salt, (+)-MK-801, and dantrolene (Tocris); and SIN-1 hydrochloride (Calbiochem, La Jolla, CA).

RESULTS

THC induces a concentration-dependent relaxation in rat isolated hepatic and mesenteric arterial segments (hepatic artery, pEC50 = 6.3 ± 0.1;E max = 96 ± 1%;n = 17; mesenteric artery, pEC50= 6.7 ± 0.1; E max = 97 ± 1%; n = 6). The effect of THC does not involve endothelial cells, because THC is equally potent at relaxing hepatic arteries (pEC50 = 6.2 ± 0.1;E max = 96 ± 2%;n = 5) and mesenteric arteries (Fig.1) without endothelium. To investigate whether THC activates capsaicin-sensitive sensory nerves, arteries were pretreated with 10 μm capsaicin for 30 min to cause desensitization and/or neurotransmitter depletion of sensory nerves. The effect of THC was tested after washout of capsaicin for 20 min. As shown in Figure 1 A,B, THC fails to relax such arteries. Because CGRP is the main vasodilator released from capsaicin-sensitive sensory nerves in rat hepatic and mesenteric arteries (Kawasaki et al., 1988; Zygmunt et al., 1999), we tested the effect of the CGRP receptor antagonist 8-37 CGRP on THC-induced vasorelaxations in these arteries. At 3 μm, 8-37 CGRP abolishes the vasorelaxations elicited by THC (Fig.1 A,B). Cannabinol, another naturally occurring cannabinoid, also causes vasorelaxation (pEC50 = 6.2 ± 0.1; E max = 96 ± 2%; n = 7), which is abolished in the presence of 8-37 CGRP or in arteries pretreated with capsaicin (Fig. 1 C). In mesenteric arteries, measurement of CGRP-like immunoreactivity provides direct evidence that THC and cannabinol release CGRP from capsaicin-sensitive sensory nerves. Thus, THC and cannabinol each release CGRP compared with basal CGRP levels (basal, 56.4 ± 2.4 fmol/mg protein; THC, 85.2 ± 7.2 fmol/mg protein; cannabinol, 86.7 ± 7.9 fmol/mg protein; p < 0.01;n = 6). When arteries had been pretreated with capsaicin for 30 min (followed by washout of capsaicin), THC and cannabinol could no longer evoke release above basal CGRP levels (THC, 57.7 ± 6.5 fmol/mg protein; cannabinol, 45.7 ± 3.5 fmol/mg protein; n = 6). Other cannabinoids, such as 11-OH-Δ9-THC, Δ9-THC-11-oic acid, and, as shown previously (Zygmunt et al., 1999), HU-210 and CP 55,940, do not produce sensory nerve-mediated vasorelaxation (Fig.2). The vasodilator effect of THC is not attributable to activation of CB1 receptors, because antagonists of this receptor do not inhibit the action of THC (Fig. 3 A,D).

Fig. 1.

View larger version:
Fig. 1.

The naturally occurring cannabinoids THC and cannabinol evoke sensory nerve-mediated relaxation of rat hepatic and mesenteric arterial segments contracted with phenylephrine (PhE). The concentration-dependent relaxations induced by THC (○) in hepatic (n = 5) (A) and mesenteric (n = 6) (B) arteries, and those induced by cannabinol (○) in hepatic arteries (n = 7) (C) are abolished in arterial segments pretreated with the sensory neurotoxin capsaicin (10 μm; ⋄;n = 5 and 4 for THC and cannabinol, respectively). The CGRP receptor antagonist 8-37 CGRP (3 μm; ▪) also prevents relaxations induced by THC (n = 5 and 6 for hepatic and mesenteric arteries, respectively) and cannabinol (n = 4). B, As shown by thetrace, THC also relaxes the mesenteric artery without endothelium. The dotted line shows the basal tension level before addition of PhE. Data are expressed as mean ± SEM.

Fig. 2.

View larger version:
Fig. 2.

The vasodilator action of THC and cannabinol is not mimicked by C11 hydroxy and carboxy derivatives of THC. In humans, THC is metabolized to 11-OH-Δ9-THC and Δ9-THC-11-oic acid (Burstein; 1999), both of which fail to relax phenylephrine (PhE)-contracted rat hepatic arteries (n = 3). The dashed line shows the basal tension level before addition of PhE. The structures of the potent CB1 and CB2 receptor agonists HU-210 and CP 55,940 are also shown; these agonists are synthetic derivatives of THC without an intact C11 methyl group. None of these compounds cause sensory nerve-mediated relaxation in the rat hepatic artery (Zygmunt et al., 1999).

Fig. 3.

View larger version:
Fig. 3.

Effects of CB1 and vanilloid receptor antagonists on sensory nerve-mediated relaxation induced by THC, cannabinol, and anandamide in rat hepatic and mesenteric arteries.A, The THC-induced vasorelaxation in hepatic arteries (○; n = 8) is not inhibited by the CB1 receptor antagonists SR141716A (300 nm; ▪; n = 5) and AM251 (30 nm; ●;n = 4). Vasorelaxations evoked by THC (○;n = 10) (B) and cannabinol (○; n = 7) (C) are not inhibited by the competitive vanilloid receptor antagonist capsazepine (3 μm; ●; n = 8 and 4 for THC and cannabinol, respectively) but are abolished by the noncompetitive vanilloid receptor antagonist ruthenium red (1 μm; ▪;n = 8 and 4 for THC and cannabinol, respectively).D, In mesenteric arteries, THC-induced relaxations (○;n = 6; same as in Fig. 1 B) are also unaffected by SR141716A (300 nm; ●;n = 4) and capsazepine (3 μm; ▪;n = 4) and are inhibited by 1 μmruthenium red (▴; n = 5). E, Anandamide-induced vasorelaxations in the absence (○;n = 5) and presence (●; n = 5) of 3 μm capsazepine or 1 μm ruthenium red (▪; n = 4). F, THC (10 μm) releases CGRP from rat hepatic arteries in the absence (n = 6; p < 0.001) but not in the presence (n = 6) of 1 μmruthenium red compared with basal CGRP release (n = 5). Data are expressed as mean ± SEM.

Activation of vanilloid receptors on sensory nerves leads to the release of CGRP and vasodilatation of rat hepatic and mesenteric arteries (Zygmunt et al., 1999). Therefore, we examined the effects of the vanilloid receptor antagonists capsazepine and ruthenium red (Szallasi and Blumberg, 1999) on relaxations induced by THC and cannabinol in these arteries. Whereas the noncompetitive vanilloid receptor antagonist ruthenium red (1 μm) abolishes the relaxation evoked by THC and cannabinol in hepatic arteries (Fig.3 B,C) and causes a substantial inhibition of the THC-induced vasorelaxation in mesenteric arteries (p < 0.0001) (Fig. 3 D), the competitive vanilloid receptor antagonist capsazepine (3 μm) is without effect (Fig. 3 B–D). In contrast to THC and cannabinol, anandamide induces vasorelaxation in the hepatic artery (pEC50 = 6.7 ± 0.1;E max = 97 ± 1%;n = 5) that is inhibited by capsazepine (p < 0.0001) (Fig. 3 E), confirming that capsazepine does indeed inhibit vanilloid receptors in this artery (Zygmunt et al., 1999). Ruthenium red (1 μm) also prevents the release of CGRP in rat hepatic arteries exposed to 10 μm THC (Fig. 3 F). The neurotoxin α-latrotoxin (1 nm), which causes vasorelaxation via release of CGRP from capsaicin-sensitive sensory nerves in rat hepatic arteries (Zygmunt et al., 1999), produces a complete relaxation in the presence of 1 μmruthenium red (E max = 95 ± 1%;n = 5), indicating that the nerves are still capable of releasing CGRP in the presence of this inhibitor.

The possibility that THC activates vanilloid receptors in a capsazepine-insensitive manner was tested in mouse isolated mesenteric arteries. THC and the vanilloid receptor agonists capsaicin and anandamide all evoke concentration-dependent relaxations in this preparation (Fig. 4 A). These agonists are active at submicromolar concentrations, with capsaicin (pEC50 = 7.8 ± 0.1;E max = 91 ± 4%;n = 4) being more potent than THC (pEC50 = 6.6 ± 0.1;E max = 89 ± 3%;n = 4) and anandamide (pEC50 = 6.4 ± 0.1; E max = 86 ± 3%; n = 4). THC (10 μm) and anandamide (10 μm) cannot relax arteries pre-exposed to 10 μm capsaicin or in the presence of 3 μm 8-37 CGRP (n = 3–4) (Fig. 4 A). As shown in Figure4 B, THC causes relaxation in mesenteric arteries from VR1−/− mice and their control littermates (VR1+/+), whereas the relaxant effects of anandamide and capsaicin are almost absent in arteries from VR1−/− mice.

Fig. 4.

View larger version:
Fig. 4.

THC elicits sensory nerve-mediated relaxation in mouse isolated mesenteric arteries via a vanilloid receptor-independent mechanism. A, Capsaicin (●), anandamide (▾), and THC (○) evoke concentration-dependent relaxations of mesenteric arterial segments from wild-type mice contracted with phenylephrine (PhE; n = 4). Traces, all from separate arterial segments, show that THC and anandamide (AEA) fail to relax arteries pretreated with capsaicin (10 μm; top traces) or in the presence of 8-37 CGRP (3 μm; bottom traces) (n = 3–4). This lack of effect of THC and anandamide is not attributable to the inability of arteries to respond to vasodilators, because CGRP and SIN-1 (a nitric oxide donor) cause complete relaxations. B, THC induces relaxations of the same magnitude in arteries from VR1 gene knock-out mice (VR1−/−; n = 5) and their control littermates (VR1+/+; n = 7). AEA (n = 6) and capsaicin (CAP; n = 4) are equally as effective as THC at relaxing arteries from VR1+/+mice, but they produce only minor relaxations in arteries from VR1−/− mice (n = 6 and 7 for AEA and CAP, respectively). As shown by the traces, THC also relaxes arteries that do not respond to either CAP or AEA, indicating that sensory nerves are functional in VR1−/− mice (E max = 72 ± 3%;n = 13). Data are expressed as mean ± SEM. The dashed lines in traces show the basal tension level before addition of PhE.

Ruthenium red is also an inhibitor of the ryanodine receptor channel present on intracellular calcium stores (Ma, 1993). The possibility that such an action of ruthenium red is responsible for its inhibition of THC-induced relaxation was therefore explored. In rat isolated hepatic arteries, neither 10 μm ryanodine nor 10 μm dantrolene, both of which inhibit the ryanodine receptor channel and caffeine-sensitive calcium stores at this concentration (Usachev et al., 1993; Chavis et al., 1996; Zhao et al., 2001), affects the THC-induced relaxation (THC, pEC50 = 6.2 ± 0.1;E max = 98 ± 1%; THC plus ryanodine, pEC50 = 6.2 ± 0.1;E max = 99 ± 1%; THC plus dantrolene, pEC50 = 6.2 ± 0.1; E max = 98 ± 1%; n = 4).

Subsequently, we examined the effect of extracellular calcium on the CGRP release evoked by THC in rat isolated mesenteric arteries. Both 10 μm THC and 10 mm caffeine release CGRP from rat mesenteric arteries when the extracellular calcium level is normal (Fig. 5 A). In the absence of extracellular calcium, THC can no longer release CGRP. However, the ability of caffeine to release CGRP is unaffected under the same conditions (Fig. 5 A), indicating that the intracellular calcium stores remain functionally intact in low extracellular calcium. The effect of 1 mm lanthanum, which is a nonselective calcium influx inhibitor, on THC-induced CGRP release was also examined. THC is unable to release CGRP in the presence of lanthanum, whereas caffeine responses are not significantly inhibited (Fig. 5 B).

Fig. 5.

View larger version:
Fig. 5.

THC-induced release of CGRP from sensory nerves in rat mesenteric arteries is dependent on calcium influx.A, THC (10 μm; n = 5) and caffeine (10 mm; n = 5) release CGRP from rat mesenteric arteries in PSS (p< 0.001 compared with basal CGRP release; n = 4). When calcium in the PSS is replaced by 10 μm EGTA, caffeine (n = 4) but not THC (n= 5) still evokes a release of CGRP (p < 0.001 compared with basal CGRP release; n = 4).B, THC (10 μm; n = 5) and caffeine (10 mm; n = 5) also release CGRP in Tris-buffer solution (p < 0.001 compared with basal CGRP release; n = 5). In the presence of 1 mm lanthanum, caffeine (n = 5) but not THC (n = 5) is able to release CGRP (p < 0.001 compared with basal CGRP release; n = 5). Data are expressed as mean ± SEM.

Influx of calcium through voltage-operated calcium channels (VOCCs) present on sensory nerves leads to neurotransmitter release (Geppetti et al., 1990; Evans et al., 1996; Lundberg, 1996; White, 1996). Therefore, we tested a mixture of L-, N-, and P/Q-type VOCC inhibitors on the vasorelaxation and release of CGRP evoked by THC. Neither relaxation nor CGRP release is inhibited by either calcicludine (L-, N-, and P-type VOCC inhibitor with IC50 values of 1–80 nm) (Schweitz et al., 1994) or nimodipine (L-type VOCC inhibitor with an IC50 value of ∼1 nm) (Godfraind et al., 1986) in combination with ω-conotoxin GVIA and ω-conotoxin MVIIC (N- and P/Q-type VOCC inhibitors with IC50 values of 1–100 nm) (Zygmunt and Hogestatt, 1993; Olivera et al., 1994;Hirota et al., 2000). Thus, the vasorelaxation induced by THC in rat hepatic arteries is unaffected by calcicludine plus ω-conotoxin GVIA plus ω-conotoxin MVIIC, each at a concentration of 100 nm(THC, pEC50 = 6.1 ± 0.1; E max = 98 ± 1%; THC plus VOCC inhibitors, pEC50 = 6.1 ± 0.1;E max = 96 ± 3%;n = 4). Furthermore, in rat mesenteric arteries, THC induces a significant CGRP release (p < 0.01) that is not different in the absence or presence of nimodipine plus ω-conotoxin GVIA plus ω-conotoxin MVIIC, each at a concentration of 100 nm (basal, 16.3 ± 4.3 fmol/mg protein; THC, 111 ± 17 fmol/mg protein; THC plus VOCC inhibitors, 117 ± 18 fmol/mg protein; n = 5).

Activation of glutamate receptors, which are present on sensory nerves (Li et al., 1997; Carlton and Coggeshall, 1999), is another possibility by which THC may cause calcium influx and subsequent neurotransmitter release. However, 3 μm MK-801 and 300 μmCNQX, inhibitors of ionotropic glutamate receptors (Castellano et al., 2001; Lerma et al., 2001), do not suppress the relaxation evoked by THC in rat mesenteric arteries (THC, pEC50 = 6.5 ± 0.1; E max = 97 ± 1%; THC plus MK-801, pEC50 = 6.7 ± 0.2;E max = 98 ± 2%; THC plus CNQX, pEC50 = 7.2 ± 0.1;E max = 100 ± 0%;n = 4). In fact, THC is more potent in the presence than in the absence of CNQX (p < 0.05). Glutamate (1 mm) does not relax mesenteric arteries, although the arterial segments respond to subsequent application of THC (n = 3).

The possibility that protein kinases mediate the THC-induced release of CGRP was also explored. We tested the effect of the nonselective protein kinase inhibitor staurosporine, which acts on both protein kinases A and C (Ruegg and Burgess, 1989), on the ability of THC and the protein kinase C activator PDBu to release CGRP in rat hepatic arteries. THC (10 μm) induces a significant and almost identical CGRP release in the absence and presence of 3 μm staurosporine (Fig.6 A). At 100 nm, staurosporine completely inhibits the CGRP release induced by 1 μm PDBu (Fig.6 B). In rat hepatic arteries, PDBu induces concentration-dependent relaxations, which are abolished by 3 μm 8-37 CGRP or by pretreatment with 10 μm capsaicin (Fig. 6 C). The vasorelaxations are also completely inhibited by 1 μm ruthenium red (Fig. 6 D). Capsazepine (3 μm) reduces the potency of PDBu (PDBu, pEC50 = 8.2 ± 0.1; PDBu plus capsazepine, pEC50 = 8.0 ± 0.1;n = 6–7; p = 0.055) and the maximal vasorelaxation induced by PDBu (PDBu,E max = 97 ± 3%; PDBu plus capsazepine, E max = 63 ± 7%;n = 6–7; p = 0.0017) (Fig.6 D). No vasorelaxation is obtained with 4α-PDBu (1–100 nm; n = 6), which does not activate protein kinase C (Blumberg, 1980).

Fig. 6.

View larger version:
Fig. 6.

The effect of THC on perivascular sensory nerves does not involve protein kinases A and C. A, THC (10 μm) evokes CGRP release in rat hepatic arteries in both the absence and presence of the nonselective protein kinase inhibitor staurosporine (3 μm; p < 0.001 compared with basal CGRP release; n = 6).B, The protein kinase C activator PDBu releases CGRP from rat hepatic arteries in the absence (p< 0.01; n = 6) but not in the presence of 100 nm staurosporine (n = 5) compared with basal CGRP release (n = 6). C, PDBu elicits concentration-dependent relaxations in rat hepatic arteries contracted with phenylephrine. However, PDBu cannot relax arteries pretreated with 10 μm capsaicin (●;n = 5) or in the presence of 3 μm8-37 CGRP (▪; n = 5). D, PDBu-induced vasorelaxations are also prevented by 1 μmruthenium red (▪; n = 5) and partially inhibited by 3 μm capsazepine (●; n= 6). For clarity, the same controls (○) are shown inC and D (n = 7). Data are expressed as mean ± SEM.

DISCUSSION

This study describes a novel effect of THC and cannabinol on capsaicin-sensitive primary sensory nerves. The effect of these cannabinoids, which are active at submicromolar concentrations, is not mediated by known cannabinoid receptors, because CB1 receptor antagonists are without inhibitory effect and, as shown previously, the CB1/CB2 receptor agonists HU-210, CP 55,940, and WIN 55,2128-372 cannot elicit capsaicin-sensitive vasorelaxation (Plane et al., 1997; Zygmunt et al., 1999). The presence of an intact C11 methyl group seems to be crucial for activity, because oxidation or lack of this methyl group results in inactive compounds, such as 11-OH-Δ9-THC, Δ9-THC-11-oic acid, HU-210, and CP 55,940. The ability of THC and cannabinol to activate sensory nerves is not related to their psychotropic activity, because the psychotropic cannabinoids 11-OH-Δ9-THC, HU-210, CP 55,940, and WIN 55,2128-372 do not evoke capsaicin-sensitive vasorelaxations. Furthermore, cannabinol, which is a weak CB1 receptor agonist and has little or no psychotropic activity (Pertwee, 1988; Rhee et al., 1997), is as potent as THC at eliciting vasorelaxation in the present study. This structure–activity relationship is also not consistent with the theory of alterations in membrane fluidity being the key activation mechanism (Pertwee, 1988).

We found that the effect of THC on sensory nerves is dependent on extracellular calcium and inhibited by the noncompetitive vanilloid receptor blocker ruthenium red (Amann and Maggi, 1991; Caterina et al., 1997). Interestingly, the endogenous cannabinoid anandamide induces calcium influx in sensory neurons via activation of vanilloid receptors (Zygmunt et al., 1999; Smart et al., 2000). A recent study shows that cannabidiol, a naturally occurring nonpsychotropic cannabinoid having anti-inflammatory properties (Srivastava et al., 1998; Malfait et al., 2000), activates vanilloid receptors on sensory neurons (Bisogno et al., 2001). However, our experiments with VR1 gene knock-out mice clearly show that the molecular target for THC is distinct from the VR1. The vanilloid receptor-like (VRL-1) channel is also expressed in sensory ganglia and displays a pharmacology similar to that of the putative THC-activated receptor/ion channel (Caterina et al., 1999). However, THC does not induce calcium transients in human embryonic kidney 293 cells expressing VRL-1 (P. M. Zygmunt and D. Julius, unpublished observations), and it is unclear whether VRL-1 is present on capsaicin-sensitive sensory neurons (Caterina et al., 1999). VR1 and VRL-1 belong to the family of transient receptor potential (TRP) ion channels, all of which are permeable to monovalent cations and calcium ions (Clapham et al., 2001). In addition to VR1 (TRPV1) and VRL-1 (TRPV2), TRPV4, TRPV5, and TRPV6 are sensitive to ruthenium red (Clapham et al., 2001; Hoenderop et al., 2001). Interestingly, TRPV4 and the recently cloned menthol receptor (TRPM8) are present on sensory nerves (Clapham et al., 2001; McKemy et al., 2002; Peier et al., 2002). Therefore, it would not be surprising if a member of the TRP ion channel family mediates the CB1/CB2receptor-independent effect of THC and cannabinol.

VOCCs represent an important calcium influx pathway in sensory neurons, and bradykinin and prostaglandin E2(PGE2) cause the release of sensory neuropeptides via activation of N-type VOCCs (Geppetti et al., 1990; Evans et al., 1996;Lundberg, 1996; White, 1996). However, inhibitors of common neuronal VOCCs are without effect on both CGRP release and vasorelaxation evoked by THC, excluding the involvement of neuronal VOCCs of the N-, L-, and P/Q type in the action of THC. Ionotropic glutamate receptors not only are present on primary sensory neurons (Carlton and Coggeshall, 1999) but also may mediate release of CGRP from such nerves (Jackson and Hargreaves, 1999). However, inhibitors of glutamate receptors did not suppress the action of THC in the present study. Instead, one of these inhibitors (CNQX), acting on non-NMDA glutamate receptors (Lerma et al., 2001), potentiated the THC-induced relaxation. Additional studies are needed to clarify the mechanism behind this effect, but one possibility could be that tonic glutamate receptor activity suppresses the THC signal pathway in sensory neurons. Indeed, activation of non-NMDA receptors can lead to a decrease in calcium influx and neurotransmitter release (Lerma et al., 2001). Metabotropic glutamate receptors are also present on sensory neurons (Li et al., 1997). Activation of these receptors releases calcium from caffeine- and ryanodine-sensitive intracellular calcium stores, which can lead to activation of protein kinase C (Chavis et al., 1996; Conn and Pin, 1997). Caffeine-induced calcium release from these stores triggers the release of CGRP (present study) and is inhibited by ryanodine or dantrolene (each at 10 μm) in rat dorsal root ganglion neurons (Usachev et al., 1993). However, the involvement of metabotropic glutamate receptors in THC-induced responses is unlikely, because glutamate could not mimic the action of THC. Also, THC, but not caffeine, was unable to release CGRP in the absence of extracellular calcium, and inhibition of the ryanodine receptor channel by ryanodine or dantrolene was without effect on THC-induced vasorelaxation. Together, these findings show that although ruthenium red is an inhibitor of VOCCs (Hamilton and Lundy, 1995; Cibulsky and Sather, 1999) and the ryanodine receptor channel (Ma, 1993), inhibition of these channels cannot explain the ability of ruthenium red to block the response to THC and cannabinol in the present study.

Protein kinases A and C are believed to play an important role in pain signaling (Malmberg et al., 1997; Cesare et al., 1999). Phorbol esters, such as PDBu and phorbol 12-myristate 13-acetate, activate protein kinase C and release substance P and CGRP from rat dorsal root ganglion neurons and skin sensory nerves (Ruegg and Burgess, 1989;Barber and Vasko, 1996; Kessler et al., 1999). In agreement with these studies, we found that PDBu triggers the release of CGRP from sensory nerves, leading to vasorelaxation. This PDBu-induced CGRP release is prevented by the protein kinase C inhibitor staurosporine. In contrast, THC does not act via protein kinase C, because its CGRP-releasing effect was unaffected by staurosporine even at a concentration 30 times higher than that used to inhibit the effect of PDBu. Protein kinase C can also sensitize sensory neurons and vanilloid receptors to inflammatory mediators (Cesare et al., 1999; Premkumar and Ahern, 2000;Vellani et al., 2001). Interestingly, we found that the competitive vanilloid receptor antagonist capsazepine produces only a small inhibition of the PDBu-induced relaxation, whereas ruthenium red completely blocks the response. This could indicate that PDBu, via a protein kinase C-dependent mechanism, activates the same ruthenium red-sensitive pathway as THC, which raises the possibility that the putative cannabinoid receptor/ion channel is affected by inflammatory mediators and phospholipase C activation. It is unlikely that the capsazepine-sensitive component of the PDBu-induced relaxation is attributable to a direct effect of PDBu on vanilloid receptors, because PDBu does not bind to VR1 (Chuang et al., 2001), and its release of CGRP was abolished by staurosporine in the present study. Staurosporine binds to and inhibits a variety of kinases, including protein kinase A (Ruegg and Burgess, 1989; Herbert et al., 1990), which has been proposed as a mediator of PGE2- and anandamide-induced enhancement of sensory neuropeptide release, possibly via phosphorylation of the vanilloid receptor (Hingtgen et al., 1995; Lopshire and Nicol, 1998; Cesare et al., 1999; De Petrocellis et al., 2001). However, the lack of effect of staurosporine on THC-induced CGRP release also excludes a role for cAMP-activated protein kinase A in this response.

The present study shows that THC and cannabinol cause release of sensory neuropeptides and vasorelaxation. Although they act on a molecular target distinct from VR1, these drugs have an effect on primary sensory nerves similar to those of capsaicin and other vanilloid receptor agonists (Szallasi and Blumberg, 1999; Zygmunt et al., 1999). These latter drugs are known to produce paradoxical analgesia via calcium influx and desensitization of sensory nerves (Szallasi and Blumberg, 1999; Urban et al., 2000). Whether such a mechanism contributes to the analgesic effects of THC remains to be determined. In conclusion, we have described a previously unknown action of THC and cannabinol on primary sensory nerves. Our findings are compatible with the existence of a novel cannabinoid receptor/ion channel, possibly belonging to the TRP ion channel family, which could be targeted by future analgesic and anti-inflammatory drugs devoid of psychotropic effects.

Footnotes

    • Received January 2, 2002.
    • Revision received March 12, 2002.
    • Accepted March 18, 2002.
  • This work was supported by the Swedish Research Council, the Swedish Society for Medical Research, the Segerfalk Foundation, and the Medical Faculty of Lund. P.M.Z. was supported by the Swedish Research Council.

  • Correspondence should be addressed to Peter Zygmunt, Department of Clinical Pharmacology, Institute of Laboratory Medicine, Lund University Hospital, SE-221 85 Lund, Sweden. E-mail:Peter.Zygmunt@klinfarm.lu.se.

REFERENCES

    1. Amann R,
    2. Maggi CA

    (1991) Ruthenium red as a capsaicin antagonist. Life Sci 49:849–856.

    1. Barber LA,
    2. Vasko MR

    (1996) Activation of protein kinase C augments peptide release from rat sensory neurons. J Neurochem 67:72–80.

    1. Bisogno T,
    2. Hanus L,
    3. De Petrocellis L,
    4. Tchilibon S,
    5. Ponde DE,
    6. Brandi I,
    7. Moriello AS,
    8. Davis JB,
    9. Mechoulam R,
    10. Di Marzo V

    (2001) Molecular targets for cannabidiol and its synthetic analogues: effect on vanilloid VR1 receptors and on the cellular uptake and enzymatic hydrolysis of anandamide. Br J Pharmacol 134:845–852.

    1. Blumberg PM

    (1980) In vitro studies on the mode of action of the phorbol esters, potent tumor promoters: part 1. Crit Rev Toxicol 8:153–197.

    1. Burstein SH

    (1999) The cannabinoid acids: nonpsychoactive derivatives with therapeutic potential. Pharmacol Ther 82:87–96.

    1. Carlton SM,
    2. Coggeshall RE

    (1999) Inflammation-induced changes in peripheral glutamate receptor populations. Brain Res 820:63–70.

    1. Castellano C,
    2. Cestari V,
    3. Ciamei A

    (2001) NMDA receptors and learning and memory processes. Curr Drug Targets 2:273–283.

    1. Caterina MJ,
    2. Schumacher MA,
    3. Tominaga M,
    4. Rosen TA,
    5. Levine JD,
    6. Julius D

    (1997) The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 389:816–824.

    1. Caterina MJ,
    2. Rosen TA,
    3. Tominaga M,
    4. Brake AJ,
    5. Julius D

    (1999) A capsaicin-receptor homologue with a high threshold for noxious heat. Nature 398:436–441.

    1. Cesare P,
    2. Moriondo A,
    3. Vellani V,
    4. McNaughton PA

    (1999) Ion channels gated by heat. Proc Natl Acad Sci USA 96:7658–7663.

    1. Chavis P,
    2. Fagni L,
    3. Lansman JB,
    4. Bockaert J

    (1996) Functional coupling between ryanodine receptors and L-type calcium channels in neurons. Nature 382:719–722.

    1. Chuang HH,
    2. Prescott ED,
    3. Kong H,
    4. Shields S,
    5. Jordt SE,
    6. Basbaum AI,
    7. Chao MV,
    8. Julius D

    (2001) Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition. Nature 411:957–962.

    1. Cibulsky SM,
    2. Sather WA

    (1999) Block by ruthenium red of cloned neuronal voltage-gated calcium channels. J Pharmacol Exp Ther 289:1447–1453.

    1. Clapham DE,
    2. Runnels LW,
    3. Strubing C

    (2001) The TRP ion channel family. Nat Rev Neurosci 2:387–396.

    1. Conn PJ,
    2. Pin JP

    (1997) Pharmacology and functions of metabotropic glutamate receptors. Annu Rev Pharmacol Toxicol 37:205–237.

    1. De Petrocellis L,
    2. Harrison S,
    3. Bisogno T,
    4. Tognetto M,
    5. Brandi I,
    6. Smith GD,
    7. Creminon C,
    8. Davis JB,
    9. Geppetti P,
    10. Di Marzo V

    (2001) The vanilloid receptor (VR1)-mediated effects of anandamide are potently enhanced by the cAMP-dependent protein kinase. J Neurochem 77:1660–1663.

    1. Evans AR,
    2. Nicol GD,
    3. Vasko MR

    (1996) Differential regulation of evoked peptide release by voltage-sensitive calcium channels in rat sensory neurons. Brain Res 712:265–273.

    1. Geppetti P,
    2. Tramontana M,
    3. Santicioli P,
    4. Del Bianco E,
    5. Giuliani S,
    6. Maggi CA

    (1990) Bradykinin-induced release of calcitonin gene-related peptide from capsaicin-sensitive nerves in guinea-pig atria: mechanism of action and calcium requirements. Neuroscience 38:687–692.

    1. Godfraind T,
    2. Miller R,
    3. Wibo M

    (1986) Calcium antagonism and calcium entry blockade. Pharmacol Rev 38:321–416.

    1. Hamilton MG,
    2. Lundy PM

    (1995) Effect of ruthenium red on voltage-sensitive Ca++ channels. J Pharmacol Exp Ther 273:940–947.

    1. Herbert JM,
    2. Seban E,
    3. Maffrand JP

    (1990) Characterization of specific binding sites for [3H]-staurosporine on various protein kinases. Biochem Biophys Res Commun 171:189–195.

    1. Hingtgen CM,
    2. Waite KJ,
    3. Vasko MR

    (1995) Prostaglandins facilitate peptide release from rat sensory neurons by activating the adenosine 3′,5′-cyclic monophosphate transduction cascade. J Neurosci 15:5411–5419.

    1. Hirota K,
    2. Kudo M,
    3. Kudo T,
    4. Matsuki A,
    5. Lambert DG

    (2000) Inhibitory effects of intravenous anaesthetic agents on K+-evoked norepinephrine and dopamine release from rat striatal slices: possible involvement of P/Q-type voltage-sensitive Ca2+ channels. Br J Anaesth 85:874–880.

    1. Hoenderop JG,
    2. Vennekens R,
    3. Muller D,
    4. Prenen J,
    5. Droogmans G,
    6. Bindels RJ,
    7. Nilius B

    (2001) Function and expression of the epithelial Ca2+ channel family: comparison of mammalian ECaC1 and 2. J Physiol (Lond) 537:747–761.

    1. Hogestatt ED,
    2. Zygmunt PM

    (2002) Cardiovascular pharmacology of anandamide. Prostaglandins Leukot Essent Fatty Acids 66:355–363.

    1. Holzer P

    (1992) Peptidergic sensory neurons in the control of vascular functions: mechanisms and significance in the cutaneous and splanchnic vascular beds. Rev Physiol Biochem Pharmacol 121:49–146.

    1. Jackson DL,
    2. Hargreaves KM

    (1999) Activation of excitatory amino acid receptors in bovine dental pulp evokes the release of iCGRP. J Dent Res 78:54–60.

    1. Kawasaki H,
    2. Takasaki K,
    3. Saito A,
    4. Goto K

    (1988) Calcitonin gene-related peptide acts as a novel vasodilator neurotransmitter in mesenteric resistance vessels of the rat. Nature 335:164–167.

    1. Kessler F,
    2. Habelt C,
    3. Averbeck B,
    4. Reeh PW,
    5. Kress M

    (1999) Heat-induced release of CGRP from isolated rat skin and effects of bradykinin and the protein kinase C activator PMA. Pain 83:289–295.

    1. Ledent C,
    2. Valverde O,
    3. Cossu G,
    4. Petitet F,
    5. Aubert JF,
    6. Beslot F,
    7. Bohme GA,
    8. Imperato A,
    9. Pedrazzini T,
    10. Roques BP,
    11. Vassart G,
    12. Fratta W,
    13. Parmentier M

    (1999) Unresponsiveness to cannabinoids and reduced addictive effects of opiates in CB1 receptor knockout mice. Science 283:401–404.

    1. Lerma J,
    2. Paternain AV,
    3. Rodriguez-Moreno A,
    4. Lopez-Garcia JC

    (2001) Molecular physiology of kainate receptors. Physiol Rev 81:971–998.

    1. Li H,
    2. Ohishi H,
    3. Kinoshita A,
    4. Shigemoto R,
    5. Nomura S,
    6. Mizuno N

    (1997) Localization of a metabotropic glutamate receptor, mGluR7, in axon terminals of presumed nociceptive, primary afferent fibers in the superficial layers of the spinal dorsal horn: an electron microscope study in the rat. Neurosci Lett 223:153–156.

    1. Lopshire JC,
    2. Nicol GD

    (1998) The cAMP transduction cascade mediates the prostaglandin E2 enhancement of the capsaicin-elicited current in rat sensory neurons: whole-cell and single-channel studies. J Neurosci 18:6081–6092.

    1. Lundberg JM

    (1996) Pharmacology of cotransmission in the autonomic nervous system: integrative aspects on amines, neuropeptides, adenosine triphosphate, amino acids and nitric oxide. Pharmacol Rev 48:113–178.

    1. Ma J

    (1993) Block by ruthenium red of the ryanodine-activated calcium release channel of skeletal muscle. J Gen Physiol 102:1031–1056.

    1. Malfait AM,
    2. Gallily R,
    3. Sumariwalla PF,
    4. Malik AS,
    5. Andreakos E,
    6. Mechoulam R,
    7. Feldmann M

    (2000) The nonpsychoactive cannabis constituent cannabidiol is an oral anti-arthritic therapeutic in murine collagen-induced arthritis. Proc Natl Acad Sci USA 97:9561–9566.

    1. Malmberg AB,
    2. Chen C,
    3. Tonegawa S,
    4. Basbaum AI

    (1997) Preserved acute pain and reduced neuropathic pain in mice lacking PKCgamma. Science 278:279–283.

    1. McKemy DD,
    2. Neuhausser WM,
    3. Julius D

    (2002) Identification of a cold receptor reveals a general role for TRP channels in thermosensation. Nature 416:52–58.

    1. Mechoulam R,
    2. Hanus L

    (2000) A historical overview of chemical research on cannabinoids. Chem Phys Lipids 108:1–13.

    1. Morisset V,
    2. Ahluwalia J,
    3. Nagy I,
    4. Urban L

    (2001) Possible mechanisms of cannabinoid-induced antinociception in the spinal cord. Eur J Pharmacol 429:93–100.

    1. Olivera BM,
    2. Miljanich GP,
    3. Ramachandran J,
    4. Adams ME

    (1994) Calcium channel diversity and neurotransmitter release: the omega-conotoxins and omega-agatoxins. Annu Rev Biochem 63:823–867.

    1. Peier AM,
    2. Moqrich A,
    3. Hergarden AC,
    4. Reeve AJ,
    5. Andersson DA,
    6. Story GM,
    7. Early TJ,
    8. Dragoni I,
    9. McIntyre P,
    10. Bevan S,
    11. Patapoutian A

    (2002) A TRP channel that senses cold stimuli and menthol. Cell 108:705–715.

    1. Pertwee RG

    (1988) The central neuropharmacology of psychotropic cannabinoids. Pharmacol Ther 36:189–261.

    1. Pertwee RG

    (2001) Cannabinoid receptors and pain. Prog Neurobiol 63:569–611.

    1. Plane F,
    2. Holland M,
    3. Waldron GJ,
    4. Garland CJ,
    5. Boyle JP

    (1997) Evidence that anandamide and EDHF act via different mechanisms in rat isolated mesenteric arteries. Br J Pharmacol 121:1509–1511.

    1. Premkumar LS,
    2. Ahern GP

    (2000) Induction of vanilloid receptor channel activity by protein kinase C. Nature 408:985–990.

    1. Rhee MH,
    2. Vogel Z,
    3. Barg J,
    4. Bayewitch M,
    5. Levy R,
    6. Hanus L,
    7. Breuer A,
    8. Mechoulam R

    (1997) Cannabinol derivatives: binding to cannabinoid receptors and inhibition of adenylylcyclase. J Med Chem 40:3228–3233.

    1. Ruegg UT,
    2. Burgess GM

    (1989) Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci 10:218–220.

    1. Schweitz H,
    2. Heurteaux C,
    3. Bois P,
    4. Moinier D,
    5. Romey G,
    6. Lazdunski M

    (1994) Calcicludine, a venom peptide of the Kunitz-type protease inhibitor family, is a potent blocker of high-threshold Ca2+ channels with a high affinity for L-type channels in cerebellar granule neurons. Proc Natl Acad Sci USA 91:878–882.

    1. Smart D,
    2. Gunthorpe MJ,
    3. Jerman JC,
    4. Nasir S,
    5. Gray J,
    6. Muir AI,
    7. Chambers JK,
    8. Randall AD,
    9. Davis JB

    (2000) The endogenous lipid anandamide is a full agonist at the human vanilloid receptor (hVR1). Br J Pharmacol 129:227–230.

    1. Srivastava MD,
    2. Srivastava BI,
    3. Brouhard B

    (1998) Delta9 tetrahydrocannabinol and cannabidiol alter cytokine production by human immune cells. Immunopharmacology 40:179–185.

    1. Szallasi A,
    2. Blumberg PM

    (1999) Vanilloid (capsaicin) receptors and mechanisms. Pharmacol Rev 51:159–212.

    1. Urban L,
    2. Campbell EA,
    3. Panesar M,
    4. Patel S,
    5. Chaudhry N,
    6. Kane S,
    7. Buchheit K,
    8. Sandells B,
    9. James IF

    (2000) In vivo pharmacology of SDZ 249–665, a novel, non-pungent capsaicin analogue. Pain 89:65–74.

    1. Usachev Y,
    2. Shmigol A,
    3. Pronchuk N,
    4. Kostyuk P,
    5. Verkhratsky A

    (1993) Caffeine-induced calcium release from internal stores in cultured rat sensory neurons. Neuroscience 57:845–859.

    1. Vellani V,
    2. Mapplebeck S,
    3. Moriondo A,
    4. Davis JB,
    5. McNaughton PA

    (2001) Protein kinase C activation potentiates gating of the vanilloid receptor VR1 by capsaicin, protons, heat and anandamide. J Physiol (Lond) 534:813–825.

    1. White DM

    (1996) Mechanism of prostaglandin E2-induced substance P release from cultured sensory neurons. Neuroscience 70:561–565.

    1. Zhao F,
    2. Li P,
    3. Chen SR,
    4. Louis CF,
    5. Fruen BR

    (2001) Dantrolene inhibition of ryanodine receptor Ca2+ release channels: molecular mechanism and isoform selectivity. J Biol Chem 276:13810–13816.

    1. Zimmer A,
    2. Zimmer AM,
    3. Hohmann AG,
    4. Herkenham M,
    5. Bonner TI

    (1999) Increased mortality, hypoactivity, and hypoalgesia in cannabinoid CB1 receptor knockout mice. Proc Natl Acad Sci USA 96:5780–5785.

    1. Zygmunt PM,
    2. Hogestatt ED

    (1993) Calcium channels at the adrenergic neuroeffector junction in the rabbit ear artery. Naunyn Schmiedebergs Arch Pharmacol 347:617–623.

    1. Zygmunt PM,
    2. Petersson J,
    3. Andersson DA,
    4. Chuang H,
    5. Sorgard M,
    6. Di Marzo V,
    7. Julius D,
    8. Hogestatt ED

    (1999) Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400:452–457.

Articles citing this article

  • Effects of targeted deletion of cannabinoid receptors CB1 and CB2 on immune competence and sensitivity to immune modulation by {Delta}9-tetrahydrocannabinol Journal of Leukocyte Biology, 1 December 2008, 84(6):1574-1584
  • TRPV2 Is Activated by Cannabidiol and Mediates CGRP Release in Cultured Rat Dorsal Root Ganglion Neurons Journal of Neuroscience, 11 June 2008, 28(24):6231-6238
  • Plant-Derived Cannabinoids Modulate the Activity of Transient Receptor Potential Channels of Ankyrin Type-1 and Melastatin Type-8 Journal of Pharmacology and Experimental Therapeutics, 1 June 2008, 325(3):1007-1015
  • The role of the endocannabinoid system in gametogenesis, implantation and early pregnancy Human Reproduction Update, 1 September 2007, 13(5):501-513
  • Cannabidiol, unlike synthetic cannabinoids, triggers activation of RBL-2H3 mast cells Journal of Leukocyte Biology, 1 June 2007, 81(6):1512-1522
  • The in Vitro and in Vivo Cardiovascular Effects of {Delta}9-Tetrahydrocannabinol in Rats Made Hypertensive by Chronic Inhibition of Nitric-Oxide Synthase Journal of Pharmacology and Experimental Therapeutics, 1 May 2007, 321(2):663-672
  • Further Characterization of the Time-Dependent Vascular Effects of {Delta}9-Tetrahydrocannabinol Journal of Pharmacology and Experimental Therapeutics, 1 April 2006, 317(1):428-438
  • Endogenous Unsaturated C18 N-Acylethanolamines Are Vanilloid Receptor (TRPV1) Agonists Journal of Biological Chemistry, 18 November 2005, 280(46):38496-38504
  • Pungent products from garlic activate the sensory ion channel TRPA1 PNAS, 23 August 2005, 102(34):12248-12252
  • Characterization of Cannabinoid Modulation of Sensory Neurotransmission in the Rat Isolated Mesenteric Arterial Bed Journal of Pharmacology and Experimental Therapeutics, 1 October 2004, 311(1):411-419
  • Haemodynamic profile and responsiveness to anandamide of TRPV1 receptor knock-out mice The Journal of Physiology, 15 July 2004, 558(2):647-657
  • Mechanisms of ADRF release from rat aortic adventitial adipose tissue American Journal of Physiology – Heart and Circulatory Physiology, 1 March 2004, 286(3):H1107-H1113
  • Selective Ligands and Cellular Effectors of a G Protein-Coupled Endothelial Cannabinoid Receptor Molecular Pharmacology, 1 March 2003, 63(3):699-705

Science/Human: Synthetic cannabinoid improves survival after severe brain injury in clinical study

  • June 18, 2012 4:04 am

In a clinical study with 97 comatose patients a synthetic cannabinoid (named KN38-7271) improved survival in the acute early phase after a head injury. The trial was conducted in 14 EURopean neurosurgical centres. KN38-7271 binds both to the CB1 and the CB2 receptor, similar to THC and some other cannabinoids. Participants received 1 or 0.5 mg of the cannabinoid or a placebo within 4.5 hours of the injury. Efficacy was measured by survival and by neurological improvement or deterioration 7 and 14 days and 1, 3, and 6 months after the injury. Intracranial pressure and cerebral perfusion pressure were analysed from start of treatment to end of day 7.

Survival rates within 1 month of the injury were significantly better in the treatment groups than in the placebo group, but this effect was not seen after 6 months. Critical intracranial pressure and cerebral perfusion pressure were less extreme and less frequent in the treatment groups. There were no serious adverse effects. Authors concluded that “KN38-7271 appeared beneficial in the acute early phase of the comatose patient after a head injury.” These results may provide the basis for further trials in larger study populations.

Klinik für Neurochirurgie, Universitätsklinikum, Otto-von-Guericke-Universität Magdeburg, Germany.

Firsching R, Piek J, Skalej M, Rohde V, Schmidt U, Striggow F; the KN38-7271 Study Group. Early Survival of Comatose Patients after Severe Traumatic Brain Injury with the Dual Cannabinoid CB1/CB2 Receptor Agonist KN38-7271: A Randomized, Double-Blind, Placebo-Controlled Phase II Trial. J Neurol Surg A Cent EUR Neurosurg. 2012 Jun 13. [in press]

Cannabinoids and their healng properties

  • June 17, 2012 6:50 pm