Carlton E. Turner

Effects of the administration of coca alkaloids on the primary immune responses of mice: Interaction with Δ9-tetrahydrocannabinol and ethanol

The effects of cocaine, its metabolites, and other alkaloids from Erythroxylon coca on the primary immune responses of ICR mice to sheep red blood cells (sRBC) and dinitrofluorobenzene (DNFB) were studied. The Jerne hemolytic plaque assay (PFC) was used to evaluate the humoral immune response to sheep red blood cells, and the delayed type hypersensitivity (DTH) response to DNFB was used to study cellular immune responsiveness. Drugs were given in single daily po doses for 5 consecutive days beginning on the day of immunization or 3 days prior to and on Days 3 and 4 after immunization. Inhibition of both PFC and DTH responses occurred at doses of 15 to 60 mg/kg of cocaine and was greatest when fed during immunization. Five other alkaloids also suppressed the PFC and/or DTH response. Cocaine was more suppressive than the six other alkaloids tested. Ethanol (5 g/kg) did not suppress the DTH response and only marginally suppressed the PFC response. delta 9-THC inhibited the PFC response at doses of 10 mg/kg and marginally suppressed the DTH response at doses of 30 mg/kg, but not at other doses ranging from 10 to 90 mg/kg. Coadministration of 5 g/kg ethanol and 15 mg/kg cocaine resulted in 50% antagonism of effects of cocaine on the PFC response and complete antagonism of the suppression of the DTH response, but only if these substances were given during the period of immunization. Like ethanol, delta 9-THC also abolished the inhibitory effects of cocaine on the PFC and DTH response but only if coadministered during the period of immunization. Coadministration of ethanol and delta 9-THC resulted in synergistic inhibition of both DTH and PFC responses.

CULTIVATION, EXTRACTION, AND ANALYSIS OF CANNABIS SATIVA L.*

Cannabis sativa L. (marijuana) has been cultivated at the University of Mississippi since 1968 under contract support from the National Institute of Mental Health. The primary objective is to provide fully defined standardized marijuana for research. Confiscated marijuana of unknown history is often adulterated. C. sutiva was grown on three sites in Mississippi in 1968 and on two sites in 1969 and 1970. The soil types were loess (clay) on campus (site A), sandy loam some 300 miles south of campus (site B), and clay loam in the delta region of the state (site C) . Site C was used only in 1968. Land was prepared by conventional agricultural methods and treated with slag at the rate of two tonslacre. Seeds were planted mechanically or by hand at depths of about one-half inch in 40-inch-wide rows with a side dressing of 13-13-13 fertilizer at the rate of 300 pounds/acre. Plantings were made in May, June, and July. Seeds germinated in less than one week if the soil was moist. Weeds were controlled by cultivators and hand hoes. No herbicides or insecticides were used. Weed control was especially important to the young plants, since their growth was greatly inhibited by competition for light, water, and soil nutrients. Plants were watered at least once a week when it did not rain. Plants were usually thinned to 12-18 inches apart when 12-24 inches high. This was best accomplished by clipping an inch above the ground, since uprooting frequently set back or killed neighboring plants. This method allowed greater branching and leaf and flower development. Plants were also side-dressed one or two additional times during the growing season. Many seed types have been planted in this study. One cannot help being amazed at the many morphological variations observed among plants produced by these seed types, and it must be concluded that Cannabis is either a genus composed of more than 100 species or a single species that has not stabilized and that has many variations. We prefer to look upon these plants as a single species, Cannabis sutiva, as first named by Linnaeus. Plants within a given seed type tended to resemble each other and grow at similar rates, although some marked differences were occasionally observed. Plants grew as rapidly as two feet a week during peak growth and were three to eighteen feet in height at maturity. Branches measured from a few inches to eight feet in length and leaves from two to twelve inches across, and the number of leaflets per leaf ranged from one to eleven, almost always odd in number (FIGURES 1, 2, and 3). Plants of several variants were observed to contain many

Biological Activity of Cannabichromene, its Homologs and Isomers

Abstract: Cannabichromene (CBC) is one of four major cannabinoids in Cannabis sativa L. and is the second most abundant cannabinoid in drug‐type cannabis. Cannabichromene and some of its homologs, analogs, and isomers were evaluated for antiinflammatory, antibacterial, and antifungal activity. Antiinflammatory activity was evaluated by the carrageenan‐induced rat paw edema and the erythrocyte membrane stabilization method. In both tests, CBC was superior to phenylbutazone. Antibacterial activity of CBC and its isomers and homologs was evaluated using gram‐positive, gram‐negative, and acid‐fast bacteria. Antifungal activity was evaluated using yeastlike and filamentous fungi and a dermatophyte. Antibacterial activity was strong, and the antifungal activity was mild to moderate.

Constituents of Erythroxylon coca II. Gas-chromatographic analysis of cocaine and other alkaloids in coca leaves☆

A gas-chromatographic procedure is outlined for the analysis of cocaine and cis- and trans-cinnamoylcocaines in coca leaves. The procedure was applied to the determination of these alkaloids in coca leaves collected from three different locations in Peru. The extraction of alkaloids from plant material was accomplished following the procedure we previously outlined. Recovery of cinnamoylcocaines was quantitative and the average coefficient of variation was 6.8%. Using androst-4-ene-3,17-dione as the internal standard, the correlation coefficient (r2) as determined from the calibration curve of trans-cinnamoylcocaine was 0.99 and the response factor 0.72. Observable differences in relative proportions of cis and trans isomers of cinnamoylcocaine were found among samples from different locations. In addition, the relative proportions of the total cinnamoylcocaine concentration to cocaine varied with sample origin. We have also examined coca extracts using a nitrogen detector and by gas chromatography--mass spectrometry. The nature and identity of other nitrogenous peaks in the chromatograms are also discussed.

Anti-inflammatory properties of cannabichromene

Abstract It was not known if Cannabichromene (CBC), which is a major constituent of drug types of Cannabis , has anti-inflammatory properties as do other cannabinoids. CBC was tested in vivo using the rat paw edema test and in vitro using the erythrocyte membrane stabilization assay. CBC was as effective as phenylbutazone (PBZ) at equivalent doses. Since CBC is less toxic than PBZ, larger doses may be given to produce a greater therapeutic effect.

The anorexic and actometric effects of cocaine and two coca extracts

The effects of cocaine and two extracts of the coca leaf were compared using locomotor activity and limited access food consumption paradigms. The three treatments were tested using both IP and PO routes of administration. The extracts were prepared by first extracting the powdered leaves with 95% ethanol, evaporating the ethanol and then partitioning the residue between water and chloroform. The doses of the extracts studied were 60, 120, 240, and 480 mg/kg. The doses of cocaine studied were 3.45, 6.9, 13.8 and 27.6 mg/kg. These doses corresponded to the amount of cocaine contained in the four doses of the chloroform layer. Cocaine and the chloroform layer (via both routes) produced dose related increases in locomotor activity and dose related decreases in food consumption. The water layer (containing only trace amounts of cocaine) produced no changes in locomotor activity; however, the highest IP dose did significantly reduce food consumption. Furthermore two of the doses (one IP, one PO) of the chloroform layer produced significantly greater effects than an equivalent amount of cocaine. These data suggest that plant constitutents other than cocaine may contribute to the overall effect achieved by chewing the leaf.

Cannabichromene and Δ9-tetrahydrocannabinol: Interactions relative to lethality, hypothermia and hexobarbital hypnosis

Abstract 1. 1. LD50s in mice after single intraperitoneal (i.p.) doses of cannabichromene (CBC) and Δ9-tetrahydrocannabinol (Δ9-THC) were 113.4 and 276.3 mg/kg, respectively. A small dose (25 mg/kg) of CBC given concurrently with Δ9-THC lowered the LD50 of Δ9-THC to 152.0 mg/kg. 2. 2. CBC, like Δ9-THC, caused hypothermia in mice; it reduced the effect of Δ9-THC at early times and increased it at later times after the two were injected simultaneously i.p. 3. 3. CBC and Δ9-THC, in 25 mg/kg i.p. doses, each prolonged hexobarbital hypnosis equally in mice, but had no additive effect in combination.

Constituents ofCannabis sativaL. XXIV: The Potency of Confiscated Marijuana, Hashish, and Hash Oil Over a Ten-Year Period

The chemical analysis of 2169 Cannabis preparations confiscated in the United States over a ten-year period is discussed. Samples are categorized according to physical appearance and potency trends are noted. The appearance of sinsemilla and buds, more potent forms of marijuana, and their effects on overall potency are emphasized along with discussion on domestically grown marijuana.

(±)9, 10-dihydroxy-Δ6a(10a)-tetrahydrocanabinol and (±)8, 9-dihydroxy-Δ6a(10a)-tetrahydrocannabinol: 2 new cannabinoids fromCannabis sativa L.

The structures of 2 new polyhydroxylated cannabinoids, (±)9, 10-dihydroxy-Δ 6a(10a)-tetrahydorcannabinol and (±)8, 9-dihydroxy-Δ 6a(10a)-tetrahydrocannabinol, obtained from a hexane extract of an IndianCannabis variant were determined by spectral means and correlation with cannabinol.

Perinatal exposure to cannabichromene and Δ9-tetrahydrocannabinol: Separate and combined effects on viability of pups and on male reproductive system at maturity

The effects of cannabichromene (CBC), delta 9-tetrahydrocannabinol (delta 9-THC) and their combination (all doses 50 mg/kg orally) were determined after being administered to female mice for 7 days beginning on the 20th day of gestation. The THC treatment reduced postnatal viability, impaired male reproductive behavior at maturity and significantly reduced seminal vesicle weights. No changes from control values occurred after CBC or CBC + THC. Thus, CBC alone at this dosage did not act like THC; moreover, it antagonized the effects of THC when the two were given in combination.