N. M. Mestechkina

Sulfated polysaccharides and their anticoagulant activity: A review

Published data on the sulfated polysaccharides of various origins that display an anticoagulant activity are summarized and analyzed. The methods used for producing semisynthetic derivatives are considered. A key role of the polysaccharide structure in the mechanisms of specific interaction with various blood plasma proteinases is discussed. The effects of the content and location of sulfate groups in polysaccharides and their molecular weight on the degree of the studied activity are assessed.

Determination of the Primary and Fine Structures of a Galactomannan from the Seed of Gleditsia triacanthos f. inermis L.

Galactomannan, a polysaccharide with a molecular weight of 660 kDa, was isolated for the first time from the seed of Gleditsia triacanthos f. inermis (yield, 15.4%). Its aqueous solutions were optically active ([α]D = +31.0°) and highly viscous ([η] = 578 ml/g). Analysis of this heteropolysaccharide using chemical, enzymatic, and chromatographic procedures, as well as IR and 13C NMR spectroscopy, showed that it consists of D-mannopyranose and D-galactopyranose residues (molar ratio, 2.42 : 1). The main chain of this galactomannan comprises 1,4-β-D-mannopyranose residues, 41% of which are substituted at C6 with single residues of α-D-galactopyranose. The probability of occurrence in the chain of mannobiose units substituted otherwise, determined experimentally, was 0.16 for the Man–Man unit, 0.50 for the Gal(Man–Man) and (Man–Man)Gal units, and 0.34 for the disubstituted Gal(Man–Man)Gal unit.

Synthesis of galactomannan sulfates

The experimental conditions for the sulfation of legume galactomannans were found, which allow for the obtainment of polysaccharides with a high degree of substitution. Sulfate esters of four galactomannans of different composition (galactose content, 16.4–47.5%) were synthesized using the SO3-pyridine complex in dimethylformamide as a sulfating agent. The degree of substitution was as high as 1.4–1.8; it did not correlate with the content of galactose in the polysaccharides. It was found that the degree of sulfation depended on the reaction temperature in the range of 19–60°C.

Anticoagulant activity of low-molecular-weight sulfated derivatives of galactomannan from Cyamopsis tetragonoloba (L.) seeds

Galactomannan from seeds of Cyamopsis tetragonoloba (L.) Taub. (guar) was depolymerized using immobilized enzymatic preparation celloviridin. A set of fragments whose molecular weights varied from 12.6 to 245.6 kDa was obtained. Sulfated derivatives of components of all fractions were synthesized, in which the content of HSO3−-groups was 48.05 ± 2.31%. All preparations exhibited anticoagulant activity, which was recorded in vitro in two tests—aIIa and aXa. The antithrombin activity (aIIa) was high (up to 65–87 U/mg) and did not depend on the molecular weight of a sulfated derivative; in the second test (aXa), the effect of molecular weight was observed. Biospecific electrophoresis allowed us to detect the ability of galactomannan sulfates to form complexes with protamine sulfate, a classic antidote to heparin.

Composition and structure of galactomannan from the seed of Gleditsia ferox Desf

Galactomannan, a heteropolysaccharide with a molecular weight of 1660 kDa, was isolated from the seed of Gleditsia ferox Desf., introduced in Russia, with a yield of 18.9%. Its aqueous solutions were optically active ([α]D = +30.5°) and highly viscous ([η] = 1430 ml/g). An analysis of the heteropolysaccharide using chemical, enzymatic, and chromatographic procedures showed that it consists of D-mannopyranose and D-galactopyranose residues (molar ratio, 2.54 : 1). The main chain of this galactomannan consists of 1,4-β-D-mannopyranose residues, 39.2% of which are substituted at C6 with single residues of α-D-galactopyranose. The probability of occurrence of mannobiose units differentially substituted with galactose was determined by 13C-NMR data and equaled, respectively, 0.37, 0.47, and 0.16 for non-substituted Man–Man units, monosubstituted Gal(Man–Man) and (Man–Man)Gal units taken together, and for the disubstituted Gal(Man–Man)Gal units.

The Structure and Characteristics of Glucomannans from Eremurus iaeand E. zangezuricus: Assignment of Acetyl Group Localization in Macromolecules

Water-soluble glucomannans from roots of Eremurus iaeand E. zangezuricuswere studied. These polysaccharides were shown to contain 28.8, 69.0, and 2.2% (E. iae) and 22.6, 74.8, and 2.6% (E. zangezuricus) D-glucose, D-mannose and acetyl groups, respectively. Their IR spectra were identical and revealed the presence of 1,4-β-glycosidic bonds and ester carbonyl groups. 13C-NMR spectroscopy revealed both polysaccharides to be linear partially acetylated 1,4-β-D-glucomannans. Acetyl groups substituted C2- and C3-hydroxyls of mannopyranose residues. A comparison of 13C-NMR data and the results of correlation analysis suggested that acetyl groups can substitute no more than one OH-group in the mannopyranosyl residue. The estimated values of [α]D, [η], and molecular weights (MW) were –34.0°, 6.05 dl/g, and 265.5 kDa, respectively, for E. iaepolysaccharide, and –38.2°, 5.4 dl/g, and 233.5 kDa, respectively, for E. zangezuricuspolysaccharide. A correlation between the intrinsic viscosities of polysaccharides and their molecular weight determined by HPLC was revealed.

Composition and structure of a galactomannan macromolecule from seeds ofAstragalus lehmannianus bunge

The composition and structure of a galactomannan from seeds ofAstragalus lehmannianus, an endemic legume species, is reported for the first time. The purified galactomannan (yield, 4.8%) contained 55% D-mannose and 45%D-galactose and had a molecular weight of 997.03 kDa. Its aqueous solutions were optically active and highly viscous (the specific rotation [α]D equaled +81.3°; the characteristic viscosity [η], 868.4 ml/g). Chemical, Chromatographic, and spectral (IR and13C-NMR spectroscopy) methods were used to demonstrate that the main chain of the molecule is formed by residues of 1,4-β-D-mannopyranose, 78% of which are substituted at position 6 with single α-D-galactopyranose. The distribution of galactose along the chain was calculated from NMR spectra: frequencies of occurrence per pair of neighboring mannose units of ( 1 ) two substituents, (2) one substituent, and (3) no substituents were 65.3%, 31.5, and 3.2, respectively. The specific rotation of the galactomannans was shown to correlate with their content of galactose.

Fractional isolation and study of the structure of galactomannan from sophora (Styphnolobium japonicum) seeds

Two fractions (1 and 2) of the galactomannan from seeds of sophora (Styphnolobium japonicum) were isolated using cold and hot aqueous extraction with a total yield of 12.88%. The two fractions differed by the ratio between mannose (Man) and galactose (Gal) residues (4.8 : 1 and 5.3 : 1, respectively) and molecular weight (1190 and 1400 kDa, respectively). Aqueous solutions of these fractions were optically active ([α]D = +4.80° and –3.36°, respectively) and highly viscous ([η] 1028.8 and 1211.2 ml/g). 13C NMR spectra of both fractions were identical with respect to the number and positions of signals, which indicates that their primary structures were identical. Using chemical and spectroscopic (IR and NMR) methods, it was shown that the galactomannan has a main chain consisting of 1,4-β-D-mannopyranose, some residues of which (16 and 17% in fractions 1 and 2, respectively) are α-galactosylated at the C-6 position. Frequencies of differently substituted mannobiose blocks in the chain, calculated for fraction 1 using NMR spectroscopic data, were 0.13 for the disubstitited blocks Gal(Man–Man)Gal, 0.37 for the sum of monosubstituted blocks Gal(Man–Man) and (Man–Man)Gal, and 0.50 for the unsubstituted block Man–Man.

Effect of polysaccharides on biological activity of human lactoferrin

The influence of neutral and ionic polysaccharides on the antioxidant (AOA) and detoxifying activities of lactoferrin (LF) and the duration of its circulation in the body was studied. In addition to natural polymers, we studied chitosan synthetic derivatives with different functional groups. On the basis of AOA test, five polysaccharides were selected. The study of the detoxifying effect of LF in two models of induced toxicity revealed polysaccharides that maintained or increased the detoxifying activity of LF. We established that the formation of a complex of lactoferrin with two galactomannans and succinyl chitosan caused positive changes in LF properties: the detoxifying activity of the protein remained unchanged or increased, whereas its elimination from the body was decelerated.

Depolymerization of legume seed galactomannan by Celloviridin G20x

The depolymerization of legume galactomannans by the commercial preparation Celloviridin G20x was studied with the aim of obtaining macromolecular fragments of a constant composition. Four galactomannans, representative of the entire range of monomer ratios characteristic of this class of phytopolysaccharides, were hydrolyzed. The galactomannans were isolated from oriental goat’s rue (Galega orientalis Lam., Man: Gal 1.1), guar (Cyamopsis tetragonoloba L., Man: Gal 1.6), honeylocust (Gleditsia triacanthos f. inermis L., Man: Gal 2.4), and sophora (Styphnolobium japonicum (L.) Schott., Man: Gal 5.1). Fragments with a monomer ratio close to that in the original polysaccharides were obtained with a high yield (75–80%). The degree of substitution of the 1,4-β-D-mannopyranose chain at position C-6 with α-galactose residues influenced the molecular weight of the final reaction product.