Melophlus sarasinorum (family Ancorinidae) is a sponge widely distributed in the Indo-West Pacific region including Great Barrier Reef, Indonesia, Palau Islands, etc. It is also known in literature as Asteropus sarasinorum, Melophlus isis and Stellettinopsis isis as junior synonymic names of the same species [11]. Moreover, erroneous names of this species were used in chemical literature such as Asteropus sarasinosum [12,13] and Melophlus sarassinorum [14]. Herein we use the valid taxonomical name.

The glycoside fraction of Melophlus sarasinorum contains complicated mixture of 14-nor-methyl-lanostane (or 30-norlanostane) triterpene glycosides. First, nine glycosides, sarasinosides A₁ (1), A₂ (2), A₃ (3), B₁ (5), B₂ (7), B₃ (9), C₁ (4), C₂ (6), and C₃ (8), were isolated and structurally elucidated by Kitagawa et al. [12,15] from the Palauan sponge. Sarasinoside A₁ (1) was independently and simultaneously isolated by Schmitz et al. [13] from the sponge collected near Guam Island and from another, Truk Lagoon collection. Other representatives of the same structural group, namely sarasinosides D (10), E (11), F (12), G (13) and H (14) along with known sarasinoside B₁ (5), were isolated by Espada et al. from the Guam Island population of the same species [16].

Lee et al. have isolated four new 14-nor-methyl-lanostane glycosides, sarasinosides H₁ (14), H₂ (15), I₁ (16) and I₂ (17) along with known sarasinosides A₁ (1) and A₃ (3) from the sponge, collected off Guam [17]. Dai et al. have isolated new 14-nor-methyl-lanostane glycosides, sarasinosides J (18), K (19), L (20) and M (21) along with known sarasinosides A₁ (1), A₃ (3), I₁ (16), I₂ (17) and H₂ (15) from M. sarasinorum collected from Sulawesi, Indonesia [14]. Finally, Santalova et al. [18] have isolated two new 14-nor-methyl-lanostane triterpene glycosides, sarasinosides A₄ (22) and A₅ (23), along with known sarasinosides A₁ (1), A₂ (2), A₃ (3), M (21) and L (20) from the Australian collection of the same species.

Most aglycones of sarasinosides have the norlanostane skeleton system with 8(9)-, 9(11)- or 8(14)-double bonds in tetracyclic moieties and identical 23-keto-Δ²⁴⁽²⁵⁾ side chains, although glycosides 2, 6 and 7 have 7(8),9(11)-diene system, while the glycosides 3, 8 and 9 possess 8(9),14-diene system. The most unusual aglycone moiety of sarasinoside D (10) contains a methyl group at C-8 instead of the usual C-14 position. The rearrangements led to relative compounds in certain triterpenoids isolated from higher plants is carried out via intermediates with Δ¹⁴⁽¹⁵⁾-unsaturation [16]. More oxidized aglycones in sarasinosides E, F, H₁, H₂, I₁, I₂, J–L and A₅ (11, 12, 14–20, 23, respectively) contain additional oxy-, methoxy- or ketone groups, while aglycones of sarasinosides M and A₄ (21, 22) are 8,9-seco-derivatives having unique 8α,9α-epoxy-8(14),9(11),24-triene and 8α,9α-epoxy-7(8),9(11)-diene structural fragments (Chart 1) [14,15,16,17,18].

Carbohydrate chains of all the glycosides are very similar to each other and have the same architecture. All sugar units (xylose, two glucoses, 2-(N-acetylamino)-2-deoxy-galactose and 2-(N-acetylamino)-2-deoxy-glucose) belong to D-series and are in pyranose forms, the configuration of all the glycosidic centers is β. Glycosides 1, 2 and 3 and many other glycosides of this structural series are pentaosides and have D-glucose as the third monosaccharide residue, while glycosides 4, 6 and 8 are tetraosides containing identical carbohydrate chains with xylose residue as the third and the terminal sugar unit.

Thus, totally 23 sarasinosides were isolated from Melophlus sarasinorum. Most of these glycosides are pentaosides and their carbohydrate chains may differ in the third monosaccharide unit (glucose or xylose) only. Structural diversity of sarasinosides is mainly provided both by oxidative processes in rings B, C and D and different positions of double bonds in the tetracyclic part of aglycones.

Data, concerning biological activities of sarasinosides are limited. Kitagawa et al. have compared ichthyotoxic action of sarasinosides A₁ (1) and B₁ (5) against killifish Poecilia reticulata [12].Glycoside 1 with glucose as the third monosaccharide unit showed LD₅₀ of 0.39 μg/mL, while glycoside 5 with xylose in this position demonstrated LD₅₀ of 0.71 μg/mL. Glycosides 1 and 5 have the same moderate inhibitory activities (ED₅₀ of 10 μg/mL) against fertilized eggs of starfish Asterina (=Patiria) pectinifera. Schmitz et al. reported the cytotoxicity of sarasinoside A₁ (1) against human lymphocytic leukemia cell line (ED₅₀ of 2.8 μg/mL) [13]. Lee et al. indicated cytotoxic activities of sarasinosides A₂ (2) and A₃ (3) against human leukemia cell line K562 (ED₅₀ of 6.5 and 17.1 μg/mL, respectively), while sarasinosides H₁ (14), H₂ (15), I₁ (16) and I₂ (17) were not active in the test [17]. Dai et al. showed that sarasinoside A₁ (1) possesses strong activity against the yeast Saccharomyces cerevisiae, but it is inactive against the bacteria Bacillus subtilis and Escherichia coli. Sarasinoside J (18) was also strongly active against S. cerevisiae, but possessed the moderate activity against B. subtilis [14]. Hence, we may conclude that sarasinosides with common 8(9)-double bond or 7(8),9(11)-diene system possess strong or moderate cytotoxic activities against tumor cell lines, yeast, fertilized eggs of starfish and ichthyotoxicity. Modifications in C and D rings, including the migration of double bond into 8(14)-position and introduction of oxygen-bearing substituents as well as other oxidative transformations in cyclic systems of aglycones decrease the activities.

Erylus formosus (the family Geodiidae) is widely distributed in shallow waters of the Caribbean Sea and the Eastern Brazil and the most studied sponge in relation of its tetracyclic triterpene glycosides. Jaspars and Crews have isolated lanostane tetraoside formoside (24) from extracts of the sponge arose during an expedition to the Bahamas [19]. The aglycone of formoside belongs to the lanostane derivatives, containing 8(9)- and 24(25)-double bonds and a carboxy group at C-14. It was earlier found from Penares sp. as a free triterpene and named penasterol [20], while its 3-O-acetyl- and 3-oxo-derivatives were also isolated from the sponge Penares incrustans [21]. All sugars (two galactoses and two arabinoses) are in pyranose forms.

As a result of studies on E. formosus, collected off the Bahamas by scuba at the depth of 55 feet, Stead et al. have isolated the related lanostane bioside eryloside F (25) with the same aglycone, also containing arabinose as the first sugar attached at C-3 of the aglycone and the second sugar unit (galactose) linked at C-2 of the arabinose residue [22].

Another glycoside of this series, formoside B (26) was found by Kubanek et al. in the sponge collected also off the Bahamas along with the known formoside (24). Glycoside 26 differs from 24 in the presence of 2-(N-acetylamino)-2-deoxy-D-galactose instead of a terminal galactose. The same group of scientists isolated also a series of fractions, containing related hexaosides and triosides and characterized these fractions by mass-spectrometry. However, individual glycoside components were not obtained [23].

A difficult task of separation of similar fractions was successfully solved by Antonov et al., who isolated a series of individual lanostane biosides, triosides and hexaosides [24,25] from the Mexican collection of the sponge. In these glycosides, all sugars are in pyranose forms, belong to D-series and have β-configuration of glycosidic bonds, except arabinose residues, which belong to L-series and have α-configuration of glycosidic bonds. Erylosides F₁–F₄ (27–30) were proved to be close related analogs of eryloside F (25) differing from each other in the aglycone side chains [23]. Previously known eryloside F (25) has also been isolated. Glycoside 27 contains the aglycone similar to penasterol with the side chain, having a methylene at C-24. Penasterol and these aglycones are predominant non-carbohydrate moieties in the glycosides from Erylus spp. Glycoside 28 has a 25(26)-double bond and a 24R-hydroxy group, while glycoside 29 contains 25(26)-double bond and 24S-hydroxy group. Eryloside F₄ (30) is a 24-keto-derivative of glycosides 28 and 29. Two new triosides, erylosides M (31) and N (32), were also isolated by this group from the same species along with eryloside H (33), earlier known from Erylus nobilis [26]. Glycoside 32 has penasterol as aglycone, while 31 contains aglycone with a methylene group at C-24 in the side chain. The previously known glycoside 33 differs from eryloside M only in the presence of 2-(N-acetylamino)-2-deoxy-D-galactose instead of a galactose residue. A new tetraoside eryloside O (34) has a similar structure to that of formoside (24), but the third terminal monosaccharide residue was identified as α-L-arabinose instead of D-galactose in formoside. New hexaosides, erylosides P (35) and Q (36), have the same carbohydrate chains and differed in the side chains of aglycones. The carbohydrate moieties of both glycosides are similar to that of formoside (24), but have an additional glucose residue attached at C-4 of the fourth sugar (galactose) and a terminal xylose residue, attached at C-2 of this glucose residue [24].

In continuation, Antonov et al. have isolated one more trioside eryloside R₁ (37) along with the known formoside (24), and six new hexaosides erylosides T₁–T₆ (38–43) [25]. All their sugars are also in pyranose forms, belong to D-series, and have β-configurations for the glycosidic bonds, except L-arabinose residues. Glycoside 37 has penasterol as aglycone, its terminal galactose is linked to C-3 of the first monosaccharide residue (arabinose), while another galactose unit is attached to C-2 of the arabinose. Hexaosides 38-40 also contain penasterol as the aglycone, while hexaosides 41–43 contain another distributed in this series aglycone with a methylene group at C-24 in side chains. All the hexaosides possess carbohydrate chains having the same general structural plan and their carbohydrate chains are similar to those of erylosides P (35) and Q (36).

It should be noted that not only hexaosides, but all the triterpene glycosides, isolated from Erylus formosus (totally 20 compounds) possess similar structural features. All of them contain the same α-L-arabinopyranose as the first monosaccharide residue, attached to C-3 in the aglycone moieties. The number of sugars may be two, three, four or six in these compounds.

One of terminal sugars is linked to C-2 of the first L-arabinose residue. The hexaosides contain also another terminal monosaccharide unit, attached to C-2 of the fourth sugar. All other linkages between monosaccharides are 1,3, except a linkage between the third fourth and the fourth monosaccharide units, which is 1,4. Not only arabinose, glucose and galactose, but also 2-(N-acetylamino)-2-deoxy-D-galactose were found as terminal monosaccharide units in the glycosides of Erylus formosus (Chart 2).

Probably, the biosynthesis of carbohydrate chains is initiated in this species by a glycosyltransferase bearing L-arabinosyl residue. Elongation of carbohydrate chains by other glycosyltransferases results in possibility of monosaccharides to be replaced by others in different triterpene glycosides of E. formosus retaining the general structural plan of carbohydrate chains [25].

Eryloside F (25) was proved to be a potent thrombin receptor antagonist, inhibiting human platelet aggregation in vitro. This glycoside facilitates the calcium mobilization in cells registered by FLIPR assay [22]. Formoside (24) shows antiviral activity (IC₅₀ of 3.5 μg/mL vs. HSV-1) and modest antibacterial action (IC₅₀ of 31.3 μg/mL vs. Corynebacterium xerosis) [19]. The fractions of hexaosides were active against amphotericin B-resistant Candida albicans (IC₅₀ of 3.9 μg/mL) [23]. Eryloside F₃ (29), but not its epimer eryloside F₂ (28) induces the early apoptosis of Ehrlich carcinoma cells at the concentration of 100 μg/mL, while eryloside F₁ (27) and eryloside F (25) activate the Ca²⁺ influx into mouse spleenocytes (130% of the control) at the same dose [24].

Erylus lendenfeldi is a species distributed in the IndoPacific geographical area. A new bis-nor-triterpene bioside eryloside A (44), containing 4β,14-di-nor-methyl-lanostane aglycone and β-D-galactopyranose residue as the first sugar residue together with another (terminal) β-D-galactopyranose residue attached to C-2 of the first monosaccharide residue in carbohydrate moiety, was isolated from the Red Sea specimen of the sponge by Carmely et al. in 1989 [27]. The aglycone of 44 contains 8(9)- and 14(15)-double bonds and a hydroxy group at C-23.

Later Sandler et al. [28] from Faulkner’s laboratory found the same glycoside in another the Red Sea collection of the sponge. In addition to 44, two new related compounds were obtained and named as erylosides K and L (45,46). The absolute configuration of the C-23 asymmetric center in 44 and45 was established through the corresponding MTPA-esters with previous hydrogenation of 45 using rhodium as a catalyst. The containing 24(25)-double bond in its aglycone eryloside K (45) was independently and almost simultaneously isolated by Fouad et al. [29] at the studies on the same sponge, collected off the Jordan coast in the Gulf of Aqaba, the Red Sea. Eryloside L (46) was structurally identified by Sandler et al. as the corresponding 23-ketone [28]. Fouad et al. isolated also similar glycoside 47, having a rare 8α,9α-epoxy-8,9-seco-7,9(11),14-triene fragment in the aglycone part (Chart 3) [29].

Bioside 47 was also named as eryloside L. Therefore, two different glycosides (46 and 47) are known now under the name “eryloside L”. We propose rename the compound 46 to eryloside L₁, because 46 was later reported.

Glycoside 44 shows cytotoxic activity against P388 tumor cell line (IC₅₀ of 4.2 μg/mL) and antifungal activity against Candida albicans (MIC of 15.6 μg/mL) [27]. This glycoside was also active against Escherichia coli, Bacillus subtilis and C. albicans (zones of inhibition at 10 μg per disc were of 6, 7 and 7 mm, respectively). Glycosides 44 and 45 lead to mortality rate of 50% at a concentration of 0.1 μg/mL in the brine shrimp assay, while compound 46 was inactive in the test. Glycosides 44, 45, and 47 did not show cytotoxicity against JURKAT, THP-1 and MM-1 tumor cell lines [29]. Glycosides 44–46 were selectively active against Δrad50 budding yeast strain (0.8, 2.0 and 3.4 μg/mL) in comparison with activity against wild parent yeast strain (IC₅₀ of 3.5, 6.1 and 11.4 μg/mL, correspondingly). It is known that selective cytotoxicity against similar mutant yeast may be an indicator of topoisomerase poisons. However, the activities against TOP1oe (IC₅₀ of 5.7, 7.5 and 10.9 μg/mL) and TOP2oe (10.8, 12.2 and 9.5 μg/mL) were milder than those of the known topoisomerase inhibitors such as camtothecin and idarubacin. The aglycone, obtained from glycoside 44, was inactive against the yeast strain that indicated a significant role of carbohydrate chain in the activity [28].

Erylus goffrilleri is an Atlantic tropical sponge species. Gulavita et al. isolated unusual lanostane glycoside, eryloside E (48), having not only a bioside carbohydrate chain consisted of galactose as the first sugar and 2-(N-acetylamino)-2-deoxy-D-glucose attached to C-2 of the galactose, but also another carbohydrate fragment composed of xylose, linked to a carboxy group at C-14 of the aglycone [30]. The aglycone has 8(9)-double bond, hydroxy-group at C-24 and additional methyl groups at C-24 and C-25 (Chart 4).

Afiyatullov et al. have isolated four related lanostane monosides, erylosides R (49), S (50), T (51) and U (52) with β-D-galactopyranose as a carbohydrate moiety [31]. Glycoside 49 contains the lanostane aglycone with 8(9)-double bond and carboxy group at C-14, hydroxy group at C-24 and two additional methyl groups at C-24 and C-25 in the side chain. Glycoside 50 contains a very similar aglycone with acetate and an additional methyl groups at C-24 in the side chain. The aglycone of glycoside 51 featured the same side chain as in 49, but has 7(8)-double bond and an uncommon lactone function between С-14 and С-9. Glycoside 52 is very similar to 51, but contains an additional 7α,8α-epoxy group.

In addition, Afiyatullov et al. have also isolated three lanostane biosides, erylosides F₅ (53), F₆ (54) and F₇ (55) and one trioside, eryloside V (56). Glycosides 53 and 54 contain closely related carbohydrate chains consisting of D-galactose and 2-(N-acetylamino)-2-deoxy-D-glucose, attached to C-2 of the galactose residue [31]. The sugars are in pyranose forms and have a β-configuration of the glycoside bonds. As it is characteristic of many erylosides, their aglycones have 8(9)-double bond and a carboxy group at C-14. The side chain of the aglycone in glycoside 53 contains a hydroxy group at C-24 and two additional methyl groups at C-24 and C-25. The side chain of 54 includes an OAc-group at C-24 and only one additional methyl group at C-24. Glycoside 55 has not only the same aglycone as in 53, but also the second sugar residue, β-D-glucopyranose, attached at C-3 of the first monosaccharide residue. Eryloside V (56) is a trioside with the same aglycon as 53 and α-L-arabinose as the first sugar. Additional alkylation in side chain is a particular structural feature of erylosides from this sponge species.

Eryloside E (48) weakly inhibited the binding of ¹²⁵[I]-Bottom Hunter labeled C5a to its receptor (IC₅₀ > 10 μM). Eryloside E shows also an immunosuppressive activity (EC₅₀ of 1.3 μg/mL), its immunosuppressive effect was specific and not caused by general cytotoxicity [30]. Erylosides R (49), S (50), T (51), V (56), F₆ (54) and F₇ (55) exhibit cytotoxic activities against Ehrlich carcinoma tumor cells (IC₅₀ of 20–40 μM) [30], while glycosides 52, 53 and 56 are inactive.

The studies on the sponge Erylus sp., isolated from a New Caledonian specimen, collected at a depth of 500 m yielded two lanostane saponins, trioside and tetraoside, named erylosides C (57) and D (58), correspondingly [32]. The aglycones of both glycosides possess 8(9)-double bond, a carboxy group at C-14, methylene group at C-24 and an additional methyl group at C-25. All the monosaccharide residues are β-D-galactopyranoses. One terminal sugar is attached to C-2 of the first monosaccharide residue, while another terminal sugar is linked to C-3 of the first sugar in the glycoside 57 as it is characteristic of many triterpene glycosides from sponges, belonging to the genus Erylus. However, one of the terminal sugars in tetraoside 58 is attached to C-4 of the second sugar, but not to C-3 (Chart 5).

The related sponge Erylus placenta was collected in the Pacific Ocean near the shore of the South Japan (Hachijo Island). Okada et al. isolated two unprecedented 14-nor-methyl-23,24,25,26,27-pentanorlanostane glycosides from the sponge. A unique aglycone of sokodoside A (59) has 8(14)-double bond, while the closely related sokodoside B (60) possesses a unprecedented conjugated 8(9),14(15),16(17)-triene system [33] (Chart 6). Glycoside 59 is a branched tetraoside. The first sugar of 59 is β-D-galactouronic acid, the second monosaccharide residue attached to C-2 of the first sugar is another residue of β-D-galactouronic acid, the third (terminal) sugar attached to C-2 of the second sugar is α-L-fucose and another terminal sugar is α-L-arabinose. Sokodoside B (60) is a branched trioside with carbohydrate chain, consisting of α-L-arabinose, β-D-galactose and β-D-galactouronic acid. In the original paper the configuration of arabinose residues was erroneously described as β, while the coupling constant in the ¹H NMR spectra of 59 and 60 definitely showed on a α-configuration for these L-arabinose residues.

Recently, a convergent synthesis of the trisaccharide moiety of sokodoside B was carried out through thioglycoside activation using sulfuric acid immobilized on silica in conjunction with N-iodosuccinimide [34]. Sokodosides A and B show moderate growth-inhibitory activities against the fungus Mortierella ramanniana and different strains of the yeast Saccharomyces cerevisiae with or without mutations (cdC₂8, act1-1 and erg6). The size of inhibition zones ranged between 8 and 16 mm at 50 μg of the tested substance on 6 mm spot on thin paper disk. Compound 59 was more active than 60. Glycosides 59 and 60 show moderate cytotoxic activities against P388 cells with IC₅₀ of 100 and 50 μg/mL, correspondingly. Hence, there is a correlation between their antifungal and cytotoxic activities.

Shin et al. have isolated four lanostane triosides erylosides G (61), H (33), I (62) and J (63) from the sponge Erylus nobilis, collected near the South shore of Jaeju Island (South Korea, Korean Strait) [26]. The aglycones of all these glycosides have 8(9)-double bond, carboxyl group at C-14 and 24-methylene group in side chains (Chart 7). Compounds 62 and 63 have also an additional methyl at C-25, like glycosides from E. goffrilleri. Both glycosides are triosides. The glycosides 61, 33, 62 and 63 demonstrate moderate cytotoxic activities against the human leukemia cell line K562 with IC₅₀ of 22.1, 24.8, 17.9 and 21.8 μg/mL, respectively.

Takada et al. have isolated nobiloside (64), a linear lanostane trioside from E. nobilis collected in the Pacific Ocean near Shikine-jima Island, 200 km southward Tokyo, Japan [35]. The aglycone of nobiloside contains 8(9)- and 24(25)-double bonds and carboxy group at C-14. The first sugar in the carbohydrate chain is β-D-glucuronic acid, the second sugar is β-D-galactouronic acid, a terminal sugar is α-L-arabinose. Synthesis of two trisaccharides, related to the saponins from E. nobilis was reported from commercially available D-galactose, L-arabinose and D-glucosamine hydrochloride via rational protecting group manipulations [36]. The glycoside shows inhibitory activity against neuraminidase from the bacterium Clostridium perfringens with IC₅₀ of 0.46 μg/mL.

Thus, all the studied Erylus species contain bioactive triterpene glycosides, in majority having 14-carboxy lanostane aglycones and carbohydrate moieties with predominance of arabinose and galactose.

The Indian Ocean sponge Ulosa sp. belongs to the family Mycalidae. Antonov et al. have isolated five glycosides with oxidized 14-nor-methyl-lanostane type aglycones, namely ulososides A–E (65–69) from the sponge collected off the waters of the North-Western Madagascar [37,38,39]. Ulososides A (65) [37], C (67), D (68) and E (69) [39] possess 22S,23R-diol fragment and 24S-methyl in side chains of their aglycones, like brassinolides. Ulososide B (66) contains a 23ξ-hydroxy group in its aglycone side chain, but has no 24-methyl group [38]. Compounds 65 is a bioside with a rare 1,6-bond between D-glucose and terminal D-glucuronic acid, while 66–68 are monosides, containing 2-(N-acetylamino)-2-deoxy-β-D-glucose or D-glucose as carbohydrate moieties. Like 65, ulososide E (69) is a bioside, but its carbohydrate chain has another monosaccharide composition with glucuronic and uncommon α-D-arabinopyranose as a terminal sugar (Chart 8).

Ectyoplasia ferox is a Carribean sponge belonging to the family Raspailliidae. Four 4β-nor-lanostane triterpenoid glycosides (70–73), containing oxidized side chains and in the ring A penasterol derivatives as aglycones have been indicated in the glycoside fraction from the sponge collected off the Bahamas [40,41]. Glycosides 70 and 71 are linear triosides, containing two β-D-galactopyranose and α-L-arabinopyranose units. Feroxosides 72 and 73 are tetraosides branched at the first monosaccharide residue with two β-D-glucopyranoses and two α-L-rhamnopyranose residues. α-L-Rhamnose units were never found in sponge triterpene tetracyclic glycosides before the study (Chart 9). Glycosides 70 and 71 showed moderate cytotoxic activities in vitro against J774 (murine monocyte-macrophage), WEHI164 (murine fibrosarcoma), and P388 (murine leukemia) cell lines at IC₅₀ ranging from 8.5 to 11.4 μg/mL [40]. Glycosides 72 and 73 were also moderately cytotoxic (IC₅₀ of 19 μg/mL) against the murine monocyte-macrophage cell line [41].

Isomalabaricane triterpene pigments, containing 4,4,8,10-tetramethyl-perhydrobenz[e]indane core with a trans-syn-trans ring junction were first reported from the Fijian collection of the sponge Jaspis stellifera (the family Ancorinidae) [42] and the Somalian sponge Stelletta sp. (the family Ancorinidae) [43]. Later they were found in several other sponges, belonging to the order Astrophorida. Some free triterpenoids of this structural group demonstrate potent antiproliferative properties and promotion of DNA polymerase β binding to DNA, but their glycosylated forms were rarely isolated. The first glycosylated isomalabarican triterpenoid, stelliferin riboside (74) with cytotoxic properties was found in unidentified species of the genus Geodia collected off Fiji Islands [44]. Recently, a new monocyclic triterpene xylopyranoside, rhabdastoside A (75), containing a skeleton related to isomalabaricanes system, was isolated along with several new isomalabaricane derivatives from the methanol extract of the sponge Rhabdastrella globostellata (the family Ancorinidae) (Chart 10) [45]. Compound 74 stabilizes DNA polymerase β binding to DNA inducing 29% binding at the concentration 28 μg/mL [46]. Two additional ribopyranosides, 13E-isomer of stelliferin riboside (76) and 3-O-deacetyl-13Z-stelliferin riboside (77), belonging to this series were isolated from another collection of R. globostellata. For their extremely high and selective activities against the mouse lymphoma L5178Y cancer cell line, the compound 76 was found to be more active than 77 with ED₅₀ values of 0.22 and 2.4 nM, respectively [47].

The sponge Asteropus sp. (the family Ancorinidae) was used by Schmitz’s group to isolate a series of triterpene galactosides, pouosides A–E (78–82) from the animals collected in Pou Bay of Truk Lagoon in 1979 [48]. It is of special interest that simultaneously they isolated sarasinosides from the same sponge [13]. Pouosides contain a new carbon skeleton of aglycones, arised from cyclization of squalene. This skeleton system is reminiscent of carotenoids of terminal cyclohexane rings linked by a symmetric acyclic chain. A cytotoxicity of pouoside A (78) was established [48].

Recently, a Korean group has found the same class of compounds in the sponge Lipastrotethya sp. (the order Halichondrida, the family Dictyonellidae) collected in Micronesia along with previously known sarasinosides. In total, they isolated nine triterpenoids, pouosides F–I (83–86) belonging to the same structural series (Chart 11) [49]. It was the unique report about the isolation of triterpene glycosides from a sponge, belonging to the order Halichondrida.

Absolute configurations of 83–86 were established by Mosher’s method. Formulae of all the pousides were corrected in accordance with these data. Some pouosides, namely pouoside A and pouosides F–I, demonstrated strong or moderate cytotoxicities against the P-388 and K562 cell lines [48,49].

The sponge Siphonochalina (=Callispongia)siphonella (the family Callispongiidae) inhabits different areas in the Red Sea. Squalene-derived triterpenes, having a new sipholane skeleton system (87) were first isolated from S. siphonella by Kashman et al. in 1981 and their structures were established using X-ray analysis [50]. About 20 compounds, having not only sipholane, but also close related siphonellane and neviotane skeleton systems were later reported [51,52]. These natural products and their semisynthetic analogs were evaluated for cytotoxicity and effect on reversing P-glycoprotein-mediated multidrug resistance to colchicin. Some of them increased the sensitivity of resistant KB-C₂ cells to the cytotoxin [53]. In majority, triterpenoids of this series present in different collections of the sponge as non-glycosylated derivatives. However, several triterpene glycosides were also found. For example, sipholenosides A and B (88,89) were isolated from S. siphonochalina, collected from the Gulf of Eilat vs. Dahlak archipelago, Red Sea. The both glycosides are α-rhamnopyranosides [54]. The originally proposed absolute configuration of sipholenol A, a main triterpene of this series was corrected by the high-field NMR application of Mosher’s method [55], herein we give the corrected structures of 88, 89.

Sipholane derivatives belong to an interesting group of triterpenoids, which attract attention not only by their structural diversity (Chart 12), but also by strong biological activities [52].

Sipholanes comprise two separate cyclic systems, obtained by two proton-initiated cyclizations from triepoxysqualenes. The precursor, suggested for their biosynthesis is (2,3S,6S,7S,18S,19S)-triepoxysqualene (91) (Scheme 1).

The sponge Xestospongia (=Neopetrosia) vanilla (the suborder Petrosina, the family Petrosiidae) inhabits underwater areas near the Californian coast-line (Canada and USA). Two first triterpene glycosides, xestovanin A and secoxestovanin A (92, 93) with new skeleton systems were found in X. vanilla by Northcote and Andersen in 1989 [56]. Aglycone moiety in 92 is formed by unusual 11,15- and 9,17-squalene cyclizations. Both xestovanin A and secoxestovanin A belong to triterpene biosides and contain carbohydrate chains with 1,2-bonded D-fucose and L-rhamnose (Chart 13). Xestovanin A (92) shows antifungal activity against Phytium ultimum. Later, the same group of scientists reported the isolation and structure elucidation of 7 new related glycosides (94–100) (Chart 14 and Chart 15). The aglycone of isoxestovanin A (94) has a new triterpene carbon skeleton, named isoxestovanine. Xestovanins B, C and dehydroxestovanin C (95, 96, 99) are triosides, containing two L-rhamnose and D-fucose residues in their carbohydrate moieties [57].

A steroidal bioside pachastrelloside A (101) from the sponge Pachastrella sp. (the family Pachastrellidae) induces the formation of multinucleated, unicellular embryos at the action on fertilized eggs of sea urchins. Pachastrelloside A has the sterol aglycone, oxidized in the rings A and B, with two monosaccharide residues (D-galactopyranosyl and 4-O-acetyl-β-D-xylopyranosyl) linked to C-4 and C-8 of its aglycone [58].

A Korean two-sponge symbiotic association was also studied. This association was composed of Poecillastra wondoensis (the family Vulcanellidae) and Jaspis wondoensis (the family Jaspidae): the upper part is P. wondoensis and the underpart is J. wondoensis. The sponge was collected at a depth of 15 m off Cheju Island, South Korea. Three steroidal glycosides, wondosterols A–C (102–104) were isolated and their stereostructures were elucidated on the basis of NMR spectroscopic analysis with application of Mosher’s method. Structures of 102–104 proved to be closely related to that of 101, although 102–104 contain only one carbohydrate chain linked to C-4 in contrast with 101 (Chart 16).Their carbohydrate moieties consist of D-galactose and D-xylose [59].

The Caribbean sponge Pandaros acanthifolium from the family Microcionidae was collected at the Canyon rock off the Martinique. The studies on the chemical composition of the sponge led to the isolation of a series of new steroid glycosides, named pandarosides. Pandarosides A–D (105–108) were structurally identified as glycosides, containing unusual sterol aglycones with a rare cis-C/D ring junction, oxidized in ring D and side chains.

Their carbohydrate moieties consist of D-glucose and D-glucuronic acids or D-glucuronic acid only at C-3 of the aglycone (Chart 17).

Methyl esters (105a, 107a and 108a) of pandarosides were also isolated. Absolute configuration of an aglycone part of 105 was established by comparison between calculated and experimental circular dichroism spectra [60]. After the isolation of additional steroidal glycosides, pandarosides E–J (109–114) and their methyl esters (109a, 111a–114a), it was shown that the majority of pandarosides exhibit in vitro activity against three of four tested parasitic protozoa. Pandaroside G (111) and its methyl ester (111a) are potent inhibitors of the growth of Trypanosoma brucei rhodesiense (IC₅₀ of 0.78 and 0.038 μM, respectively) and Leishmania donovani (IC₅₀ of 1.3 and 0.051 μM, respectively) [61]. Finally, pandarosides K–M (115–117) and methyl esters (115a–116a) were also isolated as minor components after careful chemical reinvestigation of the same species [62]. However, pandarosides K–M did not show strong antiprotozoal activities in contrast with pandaroside G.

Other steroid glycosides from the same sponge were named as acanthifoliosides A–F (118–123). Acanthifoliosides are structurally related to pandarosides steroid saponins, but they contain common steroid nuclei with trans-junction of rings C and D. Like aglycones of pandarosides, aglycones of 118–123 were oxidized in the ring D. Carbohydrate chains of the glycosides is linked to C-15 (in compounds 118–120) or to C-16 of aglycone moieties (in compounds 121–123). Acanthifoliosides A–C (118–120) were proved to be mono-β-D-xylopyranosides, acanthifoliosides D and E (121–122) are mono-α-L-rhamnopyranosides, while acanthifolioside F (123) contains a branched trioside carbohydrate chain (Chart 18). Acanthiofoliosides exhibit moderate antiprotozoal activities [63].

There is an evident structural similarity between glycosides from P. acanthifolium and steroid glycosides from starfish, because oxidation in positions 15, 16 and 23 is frequently encountered in both these groups of natural products [7,8,9].

For the first time, steroid oligoglycosides similar to asterosaponins by containing several sugar units were found in a sponge, belonging to the genera Mycale (the family Mycalidae) by our group in 1981 [64]. However, successful separation of very complicated glycoside fractions from the sponge was not achieved in that time. Twenty years later, steroid oligoglycoside mycaloside A (124) was obtained from the Caribbean sponge Mycale laxissima as individual compound [65].

In subsequent papers, structures and isolation of related mycalosides B–K (125–134) were reported [66,67]. All the glycosides are tetraosides containing the same architecture of carbohydrate chains, consisting of two D-galactopyranose, one D-glucopyranose, and one D-arabinopyranose residues. Their steroid aglycones are sterol derivatives, oxidized in rings A, D, and in side chains. Mycalosides B (125) and C (126) were shown to be 27- and 28-nor derivatives of (124), respectively. Mycaloside D (127) differs from 124 only in the presence of an additional acetyl group in the carbohydrate moiety. Mycaloside E (128) was structurally identified as a 28-nor-4-deoxy derivative of 124. Mycalosides F–H (129–131), differing from each other in structures of their side chains and nonacetylated (130, 131) or acetylated (129) tetrasaccharide carbohydrate moieties, have new 5(6)-unsaturated 3β,4β,21-trihydroxy-15-keto-steroidal aglycons. Mycaloside I (132) is a tetraoside of a new 7,24(28)-diunsaturated 3β,15β,29-trihydroxystigmastane aglycon. Mycaloside J (133) contains a new aglycone, differing from that of 126 in the absence of hydroxy group at C-4. Mycaloside K (134) is an epimer of 127 at C-24 (Chart 19). The total fraction of the mycalosides as well as purified mycalosides A (124) and G (130) inhibit the fertilization of eggs by sperm of the sea urchin Strongylocentrotus nudus preincubated with these compounds [66].

The sponge Cribrochalina olemda belongs to the family Niphatidae and inhabits tropical waters of Indo-Pacific area. Hapaioside (135) from the sponge, collected from a depth of 40 m, Micronesia, contains an unusual aglycone that resembles some steroid hormones by its polycyclic nucleus and has 4-hydroxy-6-oxo-19-norpregnane skeleton. Its sugar was identified as 6-deoxy-β-L-altropyranose-4-acetate [68]. The sugar was earlier reported as constituent of a lipopolysaccharide isolated from mammalian intestinal microorganisms.

The sponge Ptilocaulis spiculifer belongs to the family Axinellidae and inhabits the same area as C. olemda. Ptilocaulis spiculifer contains sulfated polyhydroxysteroids as characteristic secondary metabolites. Pregnane glycosides, ptilosaponosides A (136) and B (137), along with above-mentioned sulfated natural products were isolated from the sponge collected near the Solomon Islands (Chart 20). They contain β-glucose-3-sulfate as a carbohydrate moiety and have highly oxidized the ring A. These substances also have additional sulfate in the aglycones. Glycosides 136 and 137 do not show a cytotoxicity against human tumor KB cells [69]. Ptilosaponosides are exclusive steroid glycosides found in the order Halichondriidae, but not Astrophorida or Poecilosclerida.

Thus, steroid glycosides from sponges are an interesting group of natural products. Some of them demonstrate a promising antiprotozoal action and other biological activities.