Karl H. Schram

Controlled clinical trial of cannabidiol in Huntington's disease

Based on encouraging preliminary findings, cannabidiol (CBD), a major nonpsychotropic constituent of Cannabis, was evaluated for symptomatic efficacy and safety in 15 neuroleptic-free patients with Huntingtons Disease (HD). The effects of oral CBD (10 mg/kg/day for 6 weeks) and placebo (sesame oil for 6 weeks) were ascertained weekly under a double-blind, randomized cross-over design. A comparison of the effects of CBD and placebo on chorea severity and other therapeutic outcome variables, and on a Cannabis side effect inventory, clinical lab tests and other safety outcome variables, indicated no significant (p greater than 0.05) or clinically important differences. Correspondingly, plasma levels of CBD were assayed by GC/MS, and the weekly levels (mean range of 5.9 to 11.2 ng/ml) did not differ significantly over the 6 weeks of CBD administration. In summary, CBD, at an average daily dose of about 700 mg/day for 6 weeks, was neither symptomatically effective nor toxic, relative to placebo, in neuroleptic-free patients with HD.

Fast atom Bombardment Mass Spectrometry of Nucleosides, Nucleotides and Oligonucleotides

Abstract Fast atom bombardment (FAB) mass spectrometry, a new ionization technique, has been applied to a variety of polar, nonvolatile compounds with considerable success. Current literature regarding the analysis of nucleosides, nucleotides and oligonucleotides using FAB is reviewed.

The mass spectrometry of taxol.

The antitumor agent taxol has been examined by electron ionization, chemical ionization, and fast atom bombardment mass spectrometry. Three ion series are observed: (1) the M-series, characteristic of the intact molecule; (2) the T-series, with fragments derived from the taxane ring; and (3) the S-series representing the C-13 side chain. Neutral losses dominate each series of ions and serve to verify the presence and number of functionalities in each portion of the molecule. Fragmentation pathways and mechanisms of ion formation are proposed on the basis of product ion analysis and accurate mass measurements.

Assay of plasma cannabidiol by capillary gas chromatography/ion trap mass spectroscopy following high-dose repeated daily oral administration in humans

Plasma levels of cannabidiol (CBD) were ascertained weekly in 14 Huntingtons disease patients undergoing a double-blind, placebo-controlled, crossover trial of oral CBD (10 mg/kg/day = about 700 mg/day) for 6 weeks. The assay procedure involved trimethylsilyl (TMS) derivatization of CBD and the internal standard delta-6-tetrahydrocannabinol (THC), capillary column gas chromatography, ion trap mass spectroscopy in positive ion chemical ionization mode using isobutane, and calculations of CBD levels based on peak ion intensity of the 387 M + H peak of delta-6-THC-TMS and the 459 M + H peak of CBD-2TMS. The sensitivity of the assay was about 500 pg/ml, and the precision was about 10-15%. Mean plasma levels of CBD ranged from 5.9-11.2 ng/ml over the 6 weeks of CBD administration. CBD levels averaged 1.5 ng/ml one week after CBD was discontinued, and were virtually undetectable thereafter. The elimination half-life of CBD was estimated to be about 2-5 days, and there were no differences between genders for half-life or CBD levels. Additionally, no plasma delta-1-THC, the major psychoactive cannabinoid of marijuana, was detected in any subject.

Increased Eumelanin Expression and Tanning is Induced by a Superpotent Melanotropin [Nle4-d-Phe7]-α-MSH in Humans¶

Abstract Seven normal volunteers (six males and one female) with tanning skin types III or IV (Fitzpatrick scale) were given 10 daily subcutaneous injections of a superpotent synthetic analog of alpha-melanocyte stimulating hormone (α-MSH) over two weeks. This agent, [Nle4-d-Phe7]α-MSH, also called Melanotan-I (MT-I), was administered at a dose of 0.16 mg/kg/day (Monday–Friday), over a two week period. Tanning was measured serially using computerized light reflectance. This regimen induced tanning at 3 of 8 anatomic sites including the face, neck and forearm by comparison of baseline to (1) the end of the daily dosing period, (day 14), and (2) one week later, (day 21). Shave biopsies of the forearm taken at baseline and day 21 were analyzed by high performance liquid chromatography for eumelanin content which was measured as the permanganate oxidation product, pyrrole-2,3,5-tricarboxylic acid or PTCA. Pheomelanin content was measured as the hydroiodic acid digestion product, aminohydroxyphenylalanine (AHP). Eumelanin was also measured in the forehead skin samples of three subjects. The HPLC results show that mean (±SD) baseline eumelanin (PTCA) levels in forehead skin (n = 3) averaged 1.38 (±0.87) ng/mg of wet skin tissue weight. Higher mean baseline levels of PTCA were detected in forearm skin (2.06 ± 0.28 ng/mg wet weight, n = 7). One week after MT-I treatments ended, there was a mean (SD) 49% (±17.6%) increase in forehead skin PTCA levels compared to baseline (P = 0.019, n = 3, by paired sample T-test). The mean (SD) increase in forearm skin PTCA levels was 98% (±25.4%) over the same period (P = 0.003). In contrast, forearm pheomelanin expression following MT-I treatment did not significantly change from baseline. Overall, the MT-I regimen increased the eumelanin:pheomelanin ratio in forearm skin from 51:1 at baseline to 86:1 following MT-I (P = 0.054 by paired sample T-test). These results show that the tanning induced by MT-I in the face and forearm is associated with a significant increase in the eumelanin content of the human skin.

New and facile method of preparation of the anti-HIV-1 agent,1,3-dicaffeoylquinic acid

A facile and inexpensive preparation of 1,3-dicaffeoylquinic acid (cynarin) from the leaves Cynara cardunculus L. (Asteraceae) without the use any chromatographic steps is described. The procedure was based on separation of the fraction rich in 1,5-dicaffeoylquinic acid, isomerisation of 1,5-dicaffeoylquinic acid to cynarin and, owing to its higher polarity, the simple isolation of cynarin from the reaction mixture. Cynarin inhibited HIV-1 replication in MT-2 cell culture at nontoxic concentrations similar to other previously tested dicaffeoylquinic acids, which have been recently established as a potent and highly selective class of HIV-1 integrase inhibitors.

Mass spectrometry of 3,5‐ and 4,5‐dicaffeoylquinic acids and selected derivatives

The mass spectral properties of 3,5- and 4,5-dicaffeoylquinic acids (DCQAs) and selected derivatives were examined using electron ionization (EI), fast atom bombardment (FAB) and electrospray ionization (ESI). EI analysis of the trimethylsilyl derivatives provides molecular mass (M(r)) information, but the spectrum is dominated by fragment ions of the caffeic acid group; isomers cannot be differentiated using EI. FAB analysis, in both the positive and negative ion detection modes, provides M(r) information on the free compounds, but little fragmentation is observed using normal scan conditions. The FAB mass-analyzed ion kinetic energy spectroscopic analysis of the free compounds does, however, permit differentiation of the isomers, with 3,5-DCQA showing selective loss of water, a process not observed with the 4,5-isomer. Both EI and FAB provide M(r) and some structural information when applied to the peracetate derivatives of the DCQAs. ESI of the DCQAs provides considerably more structural information, especially in the negative ion detection mode, and is the recommended method of analysis of the quinic acid esters. M(r) information, identity of the ester groups and differentiation of isomers are possible using ESI. Copyright 1999 John Wiley & Sons, Ltd.

Grindelane diterpenoids from Grindelia campórum and Chrysothamnus paniculatus

Abstract The structures of nine grindelic acid related diterpenes isolated as methyl esters from Grindelia camporum and Chrysothamnus paniculatus , were elucidated based on their spectral properties.

Fast ATOM Bombardment Mass Spectra of Nucleosides. Comparison with Electron Impact and Chemical Ionization Mass Spectra

Abstract The fast atom bombardment (FAB) mass spectra of the eight major nucleosides found in RNA and DNA, pseudouridine and 2′,3′-O-isopropylidene adenosine are described and compared to El, CI, and desorption chemical ionization (DCI) spectra reported in the literature or obtained in this laboratory. Bcty, cocltl nun FAB spectra are reported. The FAB spectra are simple and provide unambiguous molecular weight information along with structurally significant fragment ions. Mechanisms of ion formation are thought to closely parallel those suggested earlier to be operating in the CI mode. Advantages and disadvantages of FAB relative to the standard ionization modes are discussed.

Analysis of Urinary Nucleosides

Conventional diagnosis, disease staging, and evaluation of therapy for cancer and acquired immune deficiency syndrome (AIDS) rely heavily on clinically observable measurements rather than biochemical tests. In the case of cancer, these measurements include tumor imaging and tissue biopsy and are accurate only after many cancer cells have been produced as a consequence of disease progression. A diagnosis of AIDS is established by detection of host-produced antibodies to the human immunodeficiency virus (HIV), whereas staging, measurement of response to therapy, and determining the type and severity of opportunistic infections (OI) suffered by AIDS patients is determined by measurement of the T- and CD-4 cell populations (1). For both cancer and AIDS, these methods can be unreliable; also, in some instances, they are painful and increase risk to the patient. For example, detection of mesothelioma (cancer of the pleural covering of the lung, most often the result of exposure to asbestos fiber) is an invasive procedure requiring rib resection to obtain tissue biopsy for cytological analysis. A dangerous level of unreliability in detecting HIV infection has been observed and results in a long lead time between HIV infection and production of antibodies. In a recent study (2), patients were shown to be infected with HIV for as long as 35 months before antibodies were detected. As a result of these deficiencies in clinical test methods, considerable research effort has been expended toward finding alternative indicators or “biomarkers” of cancer and AIDS.