Glycosides are compounds containing a carbohydrate and a noncarbohydrate residue in the same molecule. The carbohydrate residue is attached by an acetal linkage at carbon atom 1 to a noncarbohydrate residue or AGLYCONE. The nonsugar component is known as the AGLYCONE. The sugar component is called the GLYCONE. If the carbohydrate portion is glucose, the resulting compound is a GLUCOSIDE. An example is the methyl glucoside formed when a solution of glucose in boiling methyl alcohol is treated with 0.5% HCl as a catalyst.

The aglycone may be methyl alcohol, glycerol, a sterol, a phenol, etc. CLASSIFICATION OF GLYCOSIDES When the chemical nature of the aglycone group is used as the basis of systematization, the classification is:

Squalene, a triterpene, is a precursor of steroids

Many hormones are steroids Steroids

Methyl groups at C-10 and C-13 are called angular methyl groups The B, C, and D rings are trans fused

Those  on the opposite side of the plane of the ring system are  -substituents Substituents on the same side of the steroid ring system as the angular methyl groups are  -substituents The A and B rings are also trans fused in most naturally occurring steroids

Biosynthesis of Cholesterol

Synthetic Steroids

STEROIDS Steroids form an important group of compounds based on the fundamental saturated tetracyclic hydrocarbon : 1,2-cyclopentano-perhydro-phenanthrene. This nucleus, partially or completely hydrogenated, is generally substituted by methyl groups at C10 and C13. A chemical group (ketone, hydroxyl...) or an alkyl side-chain may also be present at C17. Steroids may possess a nucleus derived from sterane by one or more C-C bond scissions or ring expansions or contractions. As natural steroids are derived from squalene by cyclization, unsaturation and substitution, they may be considered as modified triterpenes.

FREE STEROLS Sterols form an important group among the steroids. Unsaturated steroids with most of the skeleton of cholestane containing a 3β-hydroxyl group and an aliphatic side chain of 8 or more carbon atoms attached to position 17 form the group of sterols. steroids In animals, another sterol, lanosterol (C30 compound) is found as a major constituent of the unsaponifiable portion of wool fat (lanoline) and was shown to be an intermediate in the biosynthesis of cholesterol. Animal tissues contain in addition to cholesterol small amounts of 7-dehydrocholesterol.

In the tissues of vertebrates, the main sterol is the C27 alcohol cholesterol (Greek, chole, bile), particularly abundant in adrenals (10%, w/w), nervous tissues (2%,w/w), liver (0.2%,w/w) and gall stones, its fundamental carbon structure being a cyclopentano-perhydro-phenanthrene ring (also called sterane). It was the first isolated sterol around 1770 by Poulletier de La Salle from gall stones. In 1815, it was isolated from the unsaponifiable fraction of animal fats and named cholesterine (Greek, khole, bile and stereos, solid). The correct formula (C27H46O) was proposed in 1888 by F. Reinitzer but structural studies from 1900 to 1932, mainly by H.O. Wieland "on the constitution of the bile acids and related substances" (Nobel Prize Chemistry 1927) and by A.O.R. Windaus on "the constitution of sterols and their connection with the vitamins" (Nobel Prize Chemistry 1928), led to the exact steric representation of cholesterol.Nobel Prize Chemistry 1927Nobel Prize Chemistry 1928 In 1936, Callow and Young designated as steroids all compounds chemically related to cholesterol.

In higher plants, the first sterols were isolated by Hesse (1878) from the Calabar beans (Phytostigma venenosum) which coined the term "phytosterine". This substance was later named stigmasterol (Windaus and Hault, 1906) from the plant genus. The denomination "phytosterol" was proposed in 1897 for all sterols of vegetal origin. Chemically, these sterols have the same basic structure as cholesterol but differences arise from the lateral chain on carbon 17. Most phytosterols are compounds having 28 to 30 carbon atoms and one or two carbon-carbon double bonds, typically one in the sterol nucleus and sometimes a second in the alkyl side chain. All phytosterols were shown to derive from cycloartenol, direct product of the cyclization of squalene in plants.

More than 200 different types of phytosterols have been reported in plant species. Representatives of these sterols are campesterol, stigmasterol and  -sitosterol which is present in all plant lipids and is used for steroid synthesis. An important C28 sterol from the fungus Claviceps purpurea (ergot) is ergosterol (mycosterol). As ergosterol is a cell membrane component largely restricted to fungi, its amount in environmental matrices may be used as an index molecule for these micro- organisms in a living biomass.

STEROL ESTERS In animal tissues, especially in the liver, adrenals and plasma lipids, cholesterol is esterified by a variety of fatty acids and most frequently by essential fatty acids, thus forming cholesterol esters. Thus, the esterification of cholesterol with arachidonic acid gives cholesteryl arachidonate. Sterol esters are important but highly variable components of the yeast cell with values ranging from traces to 50% of the total lipids. The esterification of free cholesterol within intestinal cells allows the cholesterol to be stored as a neutral lipid in cytosolic droplets and in the packing of cholesterol into lipoprotein particles for export via the plasma to liver cells.

STERYL GLYCOSIDES This family consists of one carbohydrate unit linked to the hydroxyl group of one sterol molecule. The sterol moiety was determined to be composed of various sterols: campesterol, stigmasterol, sitosterol, brassicasterol and dihydrositosterol. The sugar moiety is composed of glucose, xylose and even arabinose. In bacteria, Helicobacter was shown to be particularly rich in cholesterol glucosides (up to 33% of total lipids), thus suggesting that these molecules may be important chemotaxonomic markers for these species. It was shown that sterol glucosides participate in the synthesis of cellulose.

SAPONIN GLYCOSIDES Saponin glycosides are divided into 2 types based on the chemical structure of their aglycones (sapogenins). Saponins on hydrolysis yield an aglycone known as "sapogenin". The so-called NEUTRAL saponins are derivatives of STEROIDS with spiroketal side chains. The ACID saponins possess triterpenoid structures.

The main pathway leading to both types of sapogenins is similar and involves the head-to-tail coupling of acetate units. However, a branch occurs, after the formation of the triterpenoid hydrocarbon, squalene, that leads to steroids in one direction and to cyclic triterpenoids in the other.

Sapogenins They form the aglycon part of saponins which have well known detergent properties. They are oxygenated C27 steroids with an hydroxyl group in C-3. The structure of diosgenin is given below as an example of these compounds. These steroids can mimic or regulate steroid hormones. Thus, diosgenin can be chemically converted into corticosteroids, estrogens and progesterone. They are externally distributed in plants. They are extremely distributed in plants since they occur in over 90 plant families. They are used in nutrition, as herbal medicine, and in cosmetics.

Glycyrrhiza is the dried rhizome and roots of Glycyrrhiza glabra. Glycyrrhiza contains: 1. A saponin glycoside called Glycyrrhizin (glycyrrhizinic acid). Glycyrrhizin is the Ca 2+ and K + salts of glycyrrhizinic acid. Glycyrrhizinic acid is 50 times sweeter than sucrose. Upon hydrolysis, the glycoside loses its sweet taste and is converted to the aglycone glycyrrhetinic acid plus two molecules of glucuronic acid.

Glycyrrhetinic acid is a pentacyclic triterpenoid derivative of the beta-amyrin type. It has expectorant and antitussive properties. It is used considerably as a flavoring agent and is frequently employed to mask the taste of bitter drugs such as aloe, quinine etc. Glycyrrhetinic acid inhibits the enzymes that metabolise the prostaglandins, PGE 2 and PGF 2 to their respective 15 keto-13,14-dihydro metabolites which are INACTIVE. This causes an increased level of prostaglandins in the digestive system. Prostaglandins inhibit gastric secretion but stimulate pancreatic secretion and mucous secretion in the intestines and markedly increase intestinal motility. They also cause cell proliferation in the stomach. The effect on gastric acid secretion, promotion of mucous secretion and cell proliferation shows why licorice has potential in treating peptic ulcer. PGF 2 stimulates activity of the uterus during pregnancy and can cause abortion, therefore, licorice should not be taken during pregnancy. The structure of glycyrrhetinic acid is similar to that of cortisone. Both molecules are flat and similar at position 3 and 11.

Brassinosteroids They are derivatives of cholestane with two vicinal diols (C-2, C-3 and C-22, C-23) and a 6-keto group. They are a unique class of plant growth regulators with structural similarity to animal steroid hormones, and ecdysteroids from insects and crustacea. Many of them may be considered as sterols. The first compound isolated from Brassica napus in 1979 is brassinolide. About 40 compounds have been isolated. As it was shown that these compounds could be potent plant growth regulators, dozens of compounds of similar structure were isolated from plant sources (algae, ferns, gymnosperms, and angiosperms, but not bacteria) or synthesized

Steroid alkaloids They form a large group of molecules where a nitrogen atom is integrated into a ring or in a substituent. The steroid nucleus can contain double bonds and hydroxyls in various positions. These alkaloids are only distributed in Solanaceae (potato, tomato, eggplant...). Fortunately, their toxic properties disappear by structural transformation during ripening. Solasodine is the most common metabolite in Solanum.

Withasteroids They are C28 ergostane-type steroids with a 22,26-lactone.They are also characterized by a large number of oxygenated functions (hydroxyls, ketones, epoxides...). 90% of withasteroids (or withanolides) possess a 1-oxo-group as shown below in withaferin. Over than 200 metabolites are known, some of them as glycosides. They are predominantly associated with Solanaceae but are also found in other families (Taccacceae, Leguminosae, Labiatae). Withanolides are known to have important pharmacological properties (anti-tumor, immunosuppressive) but they are also antimicrobial, insect deterrent or ecdysteroid receptor antagonists.

Cucurbitacins They are the most oxygenated C30 triterpenoids with a dimethyl group at C-4 and methylated at C-9 and C-14. Strictly, they are not steroid since they are not methylated at C-10. The structure of cucurbitacin D is shown below as a typical example of cucurbitacins. These steroids which are commonly combined in glycosides, are mainly associated with cucurbitaceae species but they have also been detected in other families. About 50 metabolites have been identified. Mammals perceived these toxic molecules as some of the bitterest substances known. They have protective effects against herbivores but are feeding stimulants for some beetles. Cucurbitacins have been shown to act as ecdysteroid receptor antagonists.

Bile acids The end products of cholesterol utilization are the bile acids, synthesized in the liver. In mammals, the most common bile acids are C24 steroids with a carboxyl group at C-24 and up to three hydroxyl groups on the steroid nucleus, one being at C-3. The most abundant bile acids in human bile are chenodeoxycholic acid (45%) and cholic acid (31%). These are referred to as the primary bile acids. Within the intestines the primary bile acids are converted by bacteria into the secondary bile acids, identified as deoxycholate (from cholate) and lithocholate (from chenodeoxycholate). These compounds are reabsorbed by the intestines and delivered back to the liver via the portal circulation. Within the liver the carboxyl group of primary and secondary bile acids is conjugated via an amide bond to either glycine or taurine before their secretion into the bile. These conjugation reactions yield glycoconjugates and tauroconjugates, respectively. They are hydrolized in the intestine. Glycoconjugates are present only in mammals. Cholic acid : R1 = OH, R2 = H Chenodeoxycholic acid : R1 = R2 = H Glycocholic acid : R1 = OH, R2 = NH-CH 2 -COOH Taurocholic acid : R1 = OH, R2 = NH-CH 2 -CH2-SO 3 H

Cardiac Glycosides Heart diseases can be primarily grouped into three major disorders: cardiac failure, ischemia and cardiac arrhythmia. Cardiac failure can be described as the inability of the heart to pump blood effectively at a rate that meets the needs of the metabolizing tissues. This occurs when the muscles that perform contraction and force the blood out of heart are performing weakly. Thus cardiac failures primarily arise from the reduced contractility of heart muscles, especially the ventricles. Reduced contraction of heart leads to reduced heart output but new blood keeps coming in resulting in the increase in heart blood volume. The heart feels congested. Hence the term congestive heart failure. Congested heart leads to lowered blood pressure and poor renal blood flow. This results in the development of edema in the lower extremities and the lung (pulmonary edema) as well as renal failure.

Cardiac Glycosides Increasing the force of contraction of the heart (positive inotropic activity) is very important for most heart failure patients. There are several mechanisms by which this could be achieved. Cardiac steroids are perhaps the most useful. The cardiac glycosides are an important class of naturally occurring drugs whose actions include both beneficial and toxic effects on the heart. Plants containing cardiac steroids have been used as poisons and heart drugs at least since 1500 B.C. Throughout history these plants or their extracts have been variously used as arrow poisons, emetics, diuretics, and heart tonics. Cardiac steroids are widely used in the modern treatment of congestive heart failure and for treatment of atrial fibrillation and flutter. Yet their toxicity remains a serious problem.

Structure Cardiac glycosides are composed of two structural features : the sugar (glycoside) and the non-sugar (aglycone - steroid) moieties.

The R group at the 17-position defines the class of cardiac glycoside. Two classes have been observed in Nature - the cardenolides and the bufadienolides. The cardenolides have an unsaturated butyrolactone ring while the bufadienolides have an a-pyrone ring. The cardiac glycosides occur mainly in plants from which the names have been derived. Digitalis purpurea, Digitalis lanata, Strophanthus grtus, and Strophanthus kombe are the major sources of the cardiac glycosides.

The name digitoxin refers to a agent consisting of digitoxigenin (aglycone) and sugar moieties (three). The aglycone portion of cardiac glycosides is more important than the glycone portion.

The aglycone moiety: The steroid nucleus has a unique set of fused ring system that makes the aglycone moiety structurally distinct from the other more common steroid ring systems. Rings A/B and C/D are cis fused while rings B/C are trans fused. Such ring fusion give the aglycone nucleus of cardiac glycosides the characteristic U shape. The steroid nucleus has hydroxyls at 3- and 14- positions of which the sugar attachment uses the 3-OH group. 14-OH is normally unsubstituted. Many genins have OH groups at 12- and 16- positions. These additional hydroxyl groups influence the partitioning of the cardiac glycosides into the aqueous media and greatly affect the duration of action. The lactone moiety at C-17 position is an important structural feature. The size and degree of unsaturation varies with the source of the glycoside. Normally plant sources provide a 5-membered unsaturated lactone while animal sources give a 6-membered unsaturated lactone.

Sugar moiety : 1 to 4 sugars are found to be present in most cardiac glycosides attached to the 3β-OH group. The sugars most commonly used include L- rhamnose, D-glucose, D-digitoxose, D-digitalose, D-digginose, D-sarmentose, L- vallarose, and D-fructose. These sugars predominantly exist in the cardiac glycosides in the β-conformation. The presence of acetyl group on the sugar affects the lipophilic character and the kinetics of the entire glycoside. Because the order of sugars appears to have little to do with biological activity Nature has synthesized a repertoire of numerous cardiac glycosides with differing sugar skeleton but relatively few aglycone structures.

Cardenolides Their structure is closely related to bufadienolides but these C23 steroids possess a butenolide ring located at C-17. The structure of digitoxigenin is given below as a typical example of cardenolides. They are widely distributed in plants mainly as glycosides and are either toxic or insect deterrents. As potent cardiotonics, these steroids were largely studied. Monarch butterfly is well known to be highly toxic to birds because of cardenolides which come from the milkweed leaves eaten by its caterpillar.

Cardenolides (most prevalent) are C23 steroids. Cardenolides have a hormonal nature as substances. Their effects are on the heart and kidney. Strong, bitter and disagreeable taste. Cardiotonic = affect contractions of the heart muscle. Break down in fermentation by enzymatic action. Symptoms of poisoning include dizziness, vomiting, irregular heart beat, and delerium or halucinations. Treatment: atropine and activated charcoal, lidocaine Assumed mechanism of action: inhibition of the Na+, K+-ATPase resulting in increased intracellular sodium and subsequent intracellular calcium leading to enhanced muscle contraction in cardiac tissue. Cardenolides are classified according to the chemical composition of their aglycones as lanataglucosides A, B, C, D and E. Only Digitalis lanata, the woolly foxglove contains all five forms. The entire foxglove plant is toxic. Digitonin is probably the best known derivative of the Digitalis cardenolides. It is a drug derived from D. purpurea. It is used in modern medicine to increase the force of the systolic contractions and prolong duration of the diastolic phase in congestive heart failure. Digitalis drugs lower venous pressure in hypersensetive heart ailments, elevate blood pressure in a weak heart act as a diuretic, and reduce edema. However, the therapeutic dose is dangerously close to the lethal dose

Bufadienolides They are typically polyhydroxy C24 steroids with a pentadienolide ring at C-17. The structure of hellebrigenin is given below as a typical example of bufadienolides. They have been isolated from plants and animals. More than 250 compounds have been identified. In plants, thay are mostly glycosides with one to three sugars in a chain linked to the 3-hydroxyl group. They are important for their cardiotonic activity. Furthermore, they possess insecticidal and antimicrobial properties, those produced by the toad skin are strongly poisonous.

Structure - Activity Relationships The sugar moiety appears to be important only for the partitioning and kinetics of action. It possesses no biological activity. For example, elimination of the aglycone moiety eliminates the activity of alleviating symptoms associated with cardiac failure. The "backbone" U shape of the steroid nucleus appears to be very important. Structures with C/D trans fusion are inactive. Conversion to A/B trans system leads to a marked drop in activity. Thus although not mandatory A/B cis fusion is important. The 14β-OH groups is now believed to be dispensible. A skeleton without 14β-OH group but retaining the C/D cis ring fusion was found to retain activity. Lactones alone, when not attached to the steroid skeleton, are not active. Thus the activity rests in the steroid skeleton. The unsaturated 17-lactone plays an important role in receptor binding. Saturation of the lactone ring dramatically reduced the biological activity. The lactone ring is not absolutely required. For example, using a,b- unsaturated nitrile (C=C-CN group) the lactone could be replaced with little or no loss in biological activity.

Χημικές αντιδράσεις καρδιοτονωτικών γλυκοζιτών Α] (Στεροειδής σκελετός Α] (Στεροειδής σκελετός) Liebermann-Burchard AcOH+AcOAc + H 2 SO Ρόδινο---Πράσινο---Κυανό 20%SbCl 3 (CHCl 3 ) + Θέρμανση -----μώβ Β] (Λακτονικός δακτύλιος) Legal (Νιτροπρωσσικό Νάτριο) Καρδενολίδες Βαθύ ερυθρό. Βουφαδιενολίδες όχι Kedde (3,5 δίνιτρο βενζοϊκό οξύ) Καρδενολίδες ερυθριώδες Baljet ( Πικρικό οξύ) Καρδενολίδες πορτοκαλί Γ] (Σάκχαρα) Keller Killiani (Δεσόξυ-ζάχαρα) AcOH + Fe 3+ + H 2 SO κυανοπράσινη Perez (καρδιοτονωτικοί γλυκοζίτες) ΑcOH + Ξανθυδρόλη + Θέρμανση ---Ρόδινο

The larvae feed on the plant leaves for about two weeks and develop into caterpillars about 2 inches long. Monarch butterfly on milkweed Danaus plexippus Limenitis archippus