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The mass spectra of grayanotoxin III and some of its mono-, di-, and tri-ester derivatives are discussed. Some of the physiological reponses of this compound class are also presented. The extraction of this material from a natural source is discussed as well as the schemes for ester derivatization and spectral characterization of all compounds studied. High resolution mass spectral analysis and comparison of the mass spectra of similar compounds were used to define possible fragmentation mechanisms.

The (—)-∆9-trans-tetrahydrocannabinol content of Cannabis sativaL. stored at — 18, 4, and 22 ± 1 ° decomposed at a rate of 3.83, 5.38, and 6.92%, respectively, per year, whereas the material stored at 37 and 50° showed considerable decomposition. C. saticaL. stored in the absence of direct light at — 18, 4, and 22 ± 1 ° was more stable than cannabis stored under nitrogen. These data indicate that for normal research use, storage under nitrogen at 0° is not mandatory. Cannabinol is not the only decomposition product of (—)-∆9-trans-tetrahydrocannabinol. Tentative evidence supports the possible formation of hexahydrocannabinol as a decomposition product in stored C. saticaL.

The reaction of 3,5-seco-4-norpregnane-5,20-dion-3-oic acid (I) with methylamine yielded 4-methyl-4-aza-5-pregnene-3,20-dione (V) and 4-methyl-20-methylimino-4-aza-5-pregnen-3-one (VII). VII was hydrolyzed by hydrochloric acid to V. The reaction of I with ethanolamine gave 4-(β-hydroxyethyl)-4-aza-5-pregnene-3,20-dione. 5ξ-Hydroxy-4-methyl-4-azapregnane-3,20-dione (VI) was formed by the reaction of 4-oxa-5-pregnene-3,20-dione and methylamine at room temperature. VI was dehydrated to V by heating above 100 or by the use of an acid catalyst. Reduction of 4-aza-5-pregnene-3,20-dione (II), V, and VII with lithium aluminum hydride yielded 4-aza-4-pregnen-20β-ol, 4-methyl-4-aza-5-pregnen-20β-ol, and 4-methyl-20β-methylamino-4-aza-5-pregnene, respectively. By the following sequence of reactions, 4-methyl-4-aza-5-pregnen-20-one was prepared from V: ketal formation, lithium aluminum hydride reduction, and acid hydrolysis.

The synthesis of 4-dimethylaminoethyl-17α-methyl-4-aza-5-androsten-17β-ol-3-one (II) and 4-diethylaminoethyl-17α-methyl-4-aza-5-androsten-17β-ol-3-one from 17α-methyl-3, 5-seco-4-norandrostan-17β-ol-5-on-3-oic acid is described. Conditions for preparing the 5-hydroxy intermediates are reported. These lactams were converted into quaternary salts. II was reduced with lithium aluminum hydride and subsequently with hydrogen and platinum to obtain 4-dimethylaminoethyl-17α-methyl-4-aza-5α-androstan-17β-ol. Similar syntheses, beginning with 3,5-seco-4-norcholestan-5-on-3-oic acid, are described. Some of these derivatives possess antimicrobial properties.

Nine strains of Cannabis sativaL. (marijuana) were grown for research by the University of Mississippi. The seeds for these strains were obtained from Iowa, Minnesota, Mexico, Turkey, Italy, France, and Sweden. The cannabinoid content was determined using GLC, and the material was divided into two chemical phenotypes according to cannabinoid content. These phenotype categories are used to differentiate between drug-type and fiber-type Cannabis sativa.In addition, the (—)-Δ9−trans-tetrahydrocannabinol content was determined for both male and female plants, various plant parts, and a Turkish variety during various stages in its growth.

A group of nitrogen-containing steroids closely related in structure was screened for antibacterial activity, by use of Bacillus subtilisand Sarcina luteaas the test organisms. The most active compounds were cholesterol derivatives containing a tertiary or quaternary nitrogen in, or attached to, the A ring. Similar methyltestosterone or progesterone derivatives were inactive. All of the cholesterol derivatives that inhibited growth were surfactant, and, structurally, they would be classified as cationic detergents. Some of the inactive compounds were surfactant, but, structurally, they would be classified as nonionic detergents. Certain features of the antibacterial activity of one of the active steroids—ND 212 (4-dimethylaminoethyl-4-aza-5-cholesten-3-one methiodide)—were studied. Growth of a culture of B. subtiliscontaining 5 × 107cells per milliliter was inhibited by 1 μg/ml (1.7 × 10−6m) of ND 212. The amount of growth inhibition was directly related to both cell and steroid concentration. Loss of viability was rapid and irreversible. With B. subtilis, cell lysis was observed. With S. luteagrown in C14-glucose, ND 212 caused release into the media of up to 25% of the cellular radioactivity. Extensive leakage occurred before loss of viability was observed. At bacteriostatic azasteroid concentrations, there was little leakage. ND 212 was readily bound in large amounts to B. subtiliscells. Inactive azasteroids were bound poorly. C14-cholestanone was also bound, whereas C14-methyltestosterone and C14-progesterone were not bound in significant amounts. At least 50% of the bound C14-cholestanone was associated with the membrane fraction.

The synthesis of thiazolo[d-3,2]-5α-cholest-2-ene, and thiazolo[d-3,2]-17α-methyl-5α-androst-2-en-17β-ol, and eight of their 2′-derivatives is reported. These derivatives include the 2′-methyl, 2′-amino, 2′-allylamino, and 2′-phenylamino substituted thiazolosteroids. Preliminary data indicate that 2′-methylthiazolo[d-3,2]-17α methyl-5α-androst-2-en-17β-ol is a good anabolic agent.

The synthesis of some spiro-[androstane-17-y1-5′-oxazolidine] derivatives and their NMR spectra are described. Aldosterone antagonistic activity of some of the synthesized compounds is reported.

In the class of compounds known as complexes, there is a group with special properties called chelates. The name chelate is derived from the Greek word for claw, an apt descriptive term for this type of substance. A chelate is a compound formed between a metallic ion and an organic molecule having two neighboring groups capable of simultaneously combining with the metal to form a ring structure. The chelate resembles a claw with the metal ion clutched between the pincers of the organic molecule.The chelate structure has such high stability that the metal ion is no longer available for

A phytochemical investigation of an ethanolic extract of the stems and leaves of Cocculus carolinusD.C (Menispermaceae) resulted in the isolation and characterization of six compounds: the cyclitols, (+)‐quercitol and (−)‐viburnitol; the lactone, loliolide; and the alkaloids, sinoacutine, magnoflorine, and palmatine. In each case, the identity of the constituent was confirmed by spectral and mixed melting‐point comparison with authentic samples.

Smith, RodneyF. (University of Maryland, Baltimore), Donald E. Shay, and Norman J. Doorenbos. Antimicrobial action of nitrogen-containing steroids. J. Bacteriol. 85:1295–1299. 1963.—A new group of 16 synthetic nitrogen-containing steroids have been tested against a variety of microorganisms for antimicrobial properties. The gradient plate screening method, serial dilution, and dry weight techniques were used in the studies. The organisms tested consisted of 14 gram-negative bacteria, 10 gram-positive bacteria, 2 actinomycetes, 7 yeasts, and 8 molds. Inhibitory properties were found to be specific and potent in four compounds, with inhibitory concentrations as low as 0.37 μg/ml. Three of the active steroids are 4-aza cholestanes and one is a 4-nor-3,5-secocholestane amide. Sensitivity to the compounds was greatest in the gram-positive bacteria, followed by the yeasts and molds. The gram-negative bacteria were not inhibited. All 16 steroids interfered to some extent with pigmentation in Serratia marcescensbut not with pigment production in Pseudomonas aeruginosa. In a few instances, some of the molds were stimulated by the steroids at a concentration of 250 μg/ml.

The intermediate, 17α-methyl-4-aza-5α-androstan-17β-ol (III) required for the synthesis of 4-amidino-17α-methyl-4-aza-5α-androstan-17β-ol (VII) and 4-(β-guanidinoethyl)-17α-methyl-4-aza-5α-androstan-17β-ol (VI) was obtained through a reaction of 17α-methyl-3,5-seco-4-norandrostan-17β-ol-5-on-3-oic acid with ammonium hydroxide followed by two reductions (platinum dioxide and lithium aluminum hydride). Condensation of III with chloroacetonitrile under anhydrous condition, followed by reduction of the nitrile with lithium aluminum hydride, gave 4-(β-aminoethyl)-17α-methyl-4-aza-5α-androstan-17β-ol (V). The reaction of V with 2-methyl-2-thiopseudourea sulfate and of III-hydrochloride with cyanamide provided the title compounds, VI-sulfate and VII-hydrochloride, respectively. A serial dilution assay showed the title compounds to have antimicrobial activity against Saccharomyces cerevisiae.This activity is especially significant since this is the greatest activity thus far obtained in a 4-azaandrostane with a hydroxyl group at the 17β-position.

A procedure for the assay of acidic and nonacidic cannabinoids qualitatively and quantitatively by chemical fractionation prior to TLC and GLC analyses is reported. Various samples of foreign and domestic, wild and cultivated marijuana were analyzed. It was demonstrated that cannabinoids occur largely as acids, that these acids undergo decarboxylation upon storage, and that plants vary significantly in their cannabinoid composition due to heredity.

Four-week-old Cannabis sativaL. plants were treated with 1 ml. of gibberellic acid or indoleacetic acid at concentrations up to 250 p.p.m. The treatment was repeated at weekly intervals for 3 weeks. These plants were harvested 1 week after the last treatment. Gibberellic acid caused a significant increase in height and a decrease in the weight of leaves and Δ9-tetrahydrocannabinol content Indoleacetic acid produced no significant changes.

Properties of seven azasteroids and a steroidal acid, which have been found to possess antibacterial activity, were compared with benzalkonium chloride (BAC). Bactericidal action against Gaffkya tetragena, lecithin inactivation, nemolysis of sheep erythrocytes, and the lowering of surface tension were studied. The positive relationship between surface-active properties and antibacterial activity observed with many surfactants, such as BAC, was found with only two of these steroids. Certain of the bactericidal steroids may be mechanistically similar to BAC, but the properties of the steroids studied suggest different modes of action.

Synthesis of 17β-isopentyloxy-4-aza-5α-androstane was accomplished through a set of reactions involving oxidative opening of an A-ring α,β-unsaturated ketone, ring closure, and two reductions. The Williamson synthesis of ethers was employed to introduce the 17β-isopentyloxy group; ketal formation of the 3-oxo group protected the α,β-unsaturated ketone during this reaction. A serial dilution assay showed the title compound and the 4-methyl derivative to have antimicrobial activity against Gram-positive bacteria, yeasts, and molds.

3,5-Seco-4-norcholestan-5-on-3-oic acid reacts with 1,3-diaminopropane and ethylenediamine at reflux temperatures to yield 1′,4′,5′,6′-tetrahydropyrimidino-[a-4,3]-4 -aza-5-cholestene (III) and 2′,3′-dihydroimidazolino[a-4,3]-4-aza-5-cholestene (V). 1′,4′,5′,6′- Tetrahydropyrimidino[a-4,3]-17α-methyl-4-aza-5-androsten- 17β-ol (IV) and 2′,3′-dihydroimidazolino[a-4,3]- 17α-methyl-4-aza-5-androsten-17β-ol (VI) were prepared in a similar manner. The C═N bond in each of these azastroides was reduced smoothly with lithium aluminum hydride. III possesses hypocholesterolemic, anti-inflammatory, hypotensive, diuretic, antibacterial, and antifungal activities. The other derivatives are less active.

Several ring A-fused heterocyclic steroids, which were prepared as potential anabolic agents, are described. These include steroids with triazole, tetrahydropyrazine, and benzothiazine rings fused onto positions 2,3 of a derivative of methyltestosterone.

Three 4-azacholestanes and two A-norcholestanes were inactivated by 10 and 20% bovine serum and by 1.0, 2.5, and 5.0% sheep blood. The five compounds exhibited hemolytic properties when tested with 2% sheep blood and 2% human blood. These cholestanes inhibited Streptococcus pyogenesand were completely inactivated by 0.1% lecithin. Tween 80 was comparable to lecithin in causing the inactivation of steroids; 1% polyethylene glycol-4000 was inert; 1% Tween 20 and 1.0% Span 20 caused the inactivation of 3β,4-dimethyl-4-aza-5α-cholestane (ND-307). The sodium salts of four fatty acids, oleate, stearate, deoxycholate, and lauryl sulfate (0.1 to 1.0 mg/ml), effectively interfered with the action of ND-307. The steroids appear to have some properties similar to those of antimicrobial surfactants of the cationic type but have certain distinct features.

The synthesis of several types of N-substituted derivatives of 17a-aza-D-homo-5-androsten-3β-ol as potential antimicrobial agents is reported. Methyl and propyl groups were added by reductive alkylation, and β-dialkylaminoethyl and γ-dialkylaminopropyl groups were introduced by the reaction of dimethylamine or piperidine with the appropriate 17a-(ω-chloroacyl) derivative of VI. None of these derivatives exhibited significant antibacterial or antifungal activity.

6β-Methyl-4-aza-5α-cholestane (VII) and 4,6β-dimethyl-4-aza-5α-cholestane (VIII) were synthesized from 6α-methyl-4-cholesten-3-one (III) viaa sequence of reactions which included (a) ozonization of III to obtain 6α-methyl-3,5-seco-4-norcholestan-5-on-3-oic acid (IV), (b) condensation of IV with ammonium hydroxide to yield 6-methyl-4-aza-5-cholesten-3-one (V), (c) hydrogenation of V to 6β-methyl-4-aza-5α-cholestan-3-one (VI), (d) lithium aluminum hydride reduction of VI to VII, and (e) a Leuckhart methylation of VII to obtain VIII. Compounds VII and VIII possess antibacterial and antifungal properties, but are less active than 4-methyl-4-aza-5α-cholestane (I), a homolog without a 6β-methyl substituent.

Cholestenone (I) was converted into 3-aza-A-homo-4a-cholesten-4-one (II) by means of the Schmidt reaction in polyphosphoric acid. The structure was assigned on the basis of its ultraviolet spectrum, resistance to hydrolysis, and hydrogenation to a mixture of 3-aza-A-homo-5α-cholestan-4-one (IV) and 3-aza-A-homo-5α-cholestan-4-one (V). By similar procedures, progesterone (VI) yielded either 17β-acetyl-amino-4-androsten-3-one (VII) or 17β-acetylamino-3-aza-A-homo-4a-androsten-4-one (VIII), depending upon the quantity of sodium azide used. The structure of VII was confirmed by preparing it from 17β-acetylamino-5-androsten-3β-ol(IX) by Oppenauer oxidation.

The structure of euparone, a new benzofuran from Ruscus aculeatus, as 2,5-diacetyl-6-hydroxybenzofuran was determined by spectral means and conversion to euparone methyl ether (O-methyleuparone).

4,17α-Dimethyl-4-aza-5-androsten-17β-ol-3-one, 4,17α-dimethyl-4-aza-5α-androstan-17β-ol-3-one, 4,17α-dimethyl-4-aza-5α-androstan-17β-ol, and their corresponding acetate esters were synthesized and compared for antimicrobial activity. 4,17α-Dimethyl-4-aza-5α-androstan-17β-ol acetate exhibited significant activity.

An extract of Cannabis sativaL. root yielded two pentacyclic triterpenes, friedelin and epifriedelanol, and N-(p-hydroxy-β-phenylethyl)-p-hydroxy-trans-cinnamamide. Structures of the triterpenes were confirmed by preparation of derivatives and comparison to authentic friedelin. Epifriedelanol was also oxidized to friedelin. The amide was confirmed by synthesis.