Identification of Hypoglycemic Glycolipids from Ipomoea murucoides by Affinity-Directed Fractionation, In Vitro, In Silico and Dynamic Light Scattering Analysis.

In the pursuit of identifying the novel resin glycoside modulators glucose-6-phosphatase and α-glucosidase enzymes, associated with blood sugar regulation, methanol-soluble extracts from the flowers of Ipomoea murucoides (cazahuate, Nahuatl), renowned for its abundance of glycolipids, were employed. The methanol-soluble extracts were fractionated by applying the affinity-directed method with glucose-6-phosphatase enzymes from a rat's liver and α-glucosidase enzymes from its intestines. Mass spectrometry and nuclear magnetic resonance were employed to identify the high-affinity compound as a free ligand following the release from the enzymatic complex. Gel permeation through a spin size-exclusion column allowed the separated high-affinity molecules to bind to glucose-6-phosphatase and α-glucosidase enzymes in solution, which led to the identification of some previously reported resin glycosides in the flowers of cazahuate, where a glycolipid mainly structurally related to murucoidin XIV was observed. In vitro studies demonstrated the modulating properties of resin glycosides on the glucose-6-phosphatase enzyme. Dynamic light scattering revealed conformational variations induced by resin glycosides on α-glucosidase enzyme, causing them to become more compact, akin to observations with the positive control, acarbose. These findings suggest that resin glycosides may serve as a potential source for phytotherapeutic agents with antihyperglycemic properties.

Phytochemical Screening and Isolation of New Ent-Clerodane Diterpenoids from Croton guatemalensis Lotsy.

Phytochemical screening of an ethanol-water extract (EWE) from the bark of Croton guatemalensis led to the isolation and identification of eight compounds, among them: five ent-clerodane diterpenoids [junceic acid (1), 6(s)-acetoxy-15,16-diepoxy-ent-cleroda-3,13(16),14-trien-20-oic acid (crotoguatenoic acid A) (2), 6(s)-hydroxyoxy-15,16-diepoxy-ent-cleroda-3,13(16),14-trien-20-oic acid (crotoguatenoic acid B) (3), formosin F (4), bartsiifolic acid (5)], and three flavonoids [rutin (6), epicatechin (7), and quercetin (8)]. Of these, 2 and 3 are reported here for the first time. Structures were established through conventional spectroscopy methods and their absolute configurations were determined by optical rotation and comparison of experimental electronic circular dichroism (ECD) and theoretical calculated ECD spectra. A suitable high performance liquid chromatography (HPLC) method for quantifying rutin (6) was developed and validated according to standard protocols. Affinity-directed fractionation was used to identify possible in vitro active compounds on α-glucosidases from Saccharomyces cerevisiae. HPLC-ESI-MS was used to identify the inhibitors as free ligands after being released from the enzymatic complex by denaturing acidic conditions. The affinity studies led to the identification of ent-clerodane diterpenoids as active compounds. In silico analysis allowed us to determine the best conformational rearrangement for the α-glucosidase inhibitors.

Phytochemical Study of Eryngium cymosum F. Delaroche and the Inhibitory Capacity of Its Main Compounds on Two Glucose-Producing Pathway Enzymes.

One undescribed acylated flavonol glucoside and five known compounds were isolated from the aerial parts of Eryngium cymosum F. Delaroche, a plant that is used in traditional Mexican medicine to treat type 2 diabetes. The chemical structures of the isolated compounds were elucidated using a variety of spectroscopic techniques, including 1D and 2D nuclear magnetic resonance (NMR) and mass spectrometry (MS). Chlorogenic acid (1), rosmarinic acid (2), caffeic acid (3), protocatechuic acid (4), kaempferol-3-O-(2,6-di-O-trans-ρ-coumaryl)-ß-d-glucopyranoside (5), and the new acylated flavonol glucoside quercetin-3-O-(2,6-di-O-trans-ρ-coumaryl)-ß-d-glucopyranoside (6) were isolated. This is the first report on the natural occurrence of quercetin-3-O-(2,6-di-O-trans-ρ-coumaryl)-ß-D-glucopyranoside (6). In addition, according to the HPLC profile obtained for the water extract (WE), chlorogenic acid (1) and rosmarinic acid (2) were identified as the main compounds, while kaempferol-3-O-(2,6-di-O-trans-ρ-coumaryl)-ß-d-glucopyranoside (5) were the main compound in the butanolic extract. We demonstrate the important role of compound 5 over the inhibition of G6Pase and FBPase. The isolated compounds may play an important role in the hypoglycemic effect of the extract and may act in a synergic way, but more experiments are needed to corroborate these findings.

Hypoglycemic Effect of Two Mexican Medicinal Plants.

Type 2 diabetes is a worldwide prevalent disease that is due to a progressive loss of adequate ß-cell insulin secretion, frequently against a background of insulin resistance. In Mexican traditional medicine, the therapeutic use of hypoglycemic plants to control the disease is a common practice among type 2 diabetic patients. In the present work, we examined the traditional use of the aerial parts of Eryngium longifolium and the rhizome of Alsophila firma, consumed by people use over the day (in fasting state) to control their blood glucose levels, therefore, we aimed to assess the acute hypoglycemic effect of both plants. First, basic phytochemical profiles of both plants were determined and, subsequently, acute toxicity tests were carried out. Then, in vivo hypoglycemic tests were performed in streptozotocin-nicotinamide (STZ-NA) induced hyperglycemic Wistar rats and finally the effect of the plants on three enzymes involved in glucose metabolism was assayed in vitro. Through HPLC-DAD chromatography, caffeic acid, chlorogenic acid, rosmarinic acid, isoflavones, and glycosylated flavonoids were identified in E. longifolium, while the possible presence of flavanones or dihydroflavonols was reported in A. firma. Both plants exhibited a statistically significant hypoglycemic effect, without a dose-dependent effect. Furthermore, they inhibited glucose 6-phosphatase and fructose 1,6-bisphosphatase in in vitro assays, which could be associated with the hypoglycemic effect in vivo. Thus, this study confirmed for the first time the traditional use of the aerial part of E. longifolium and the rhizome of A. firma as hypoglycemic agents in a hyperglycemic animal model. In addition, it was concluded that their ability to regulate hyperglycemia could involve the inhibition of hepatic glucose output, which mainly controls glucose levels in the fasting state.

Corrigendum to "Phytochemical Composition and Chronic Hypoglycemic Effect of Bromelia karatas on STZ-NA-Induced Diabetic Rats".

[This corrects the article DOI: 10.1155/2019/9276953.].

Contribution of fasting and postprandial glucose-lowering mechanisms to the acute hypoglycemic effect of traditionally used Eryngium cymosum F.Delaroche.

ETHNOPHARMACOLOGICAL RELEVANCE: Eryngium cymosum F. Delaroche was detected as a traditional remedy against type 2 diabetes consumed by patients of Tlanchinol in the state of Hidalgo, Mexico. AIM OF THE STUDY: Assessing the hypoglycemic effect and safety of the traditional extract of E. cymosum and relating it to key glucose-lowering mechanisms both in fasting and postprandial state. MATERIALS AND METHODS: The aqueous extract of E. cymosum was subjected to HPLC analysis to identify its main components. Hyperglycaemic STZ-NA Wistar rats were administered with the extract to evaluate its effect on blood glucose levels and a possible dose-dependence. Afterward, it was evaluated in both pyruvate and maltose tolerance tests in STZ-NA rats to characterize its effect on gluconeogenesis and carbohydrate breakdown, two of the main mechanisms responsible for fasting and postprandial hyperglycaemia in type 2 diabetes patients. In addition, the inhibitory capacity of the extract was evaluated on key enzymes involved in gluconeogenesis and a-glucosidases. Moreover, insulin concentrations were measured in normoglycemic rats in both conditions to establish a link between the hypoglycaemic effect of the extract with insulin release and functioning. RESULTS: Caffeic acid (1), chlorogenic acid (2), and rosmarinic acid (3) were identified as the main constituents of the aqueous extract of E. cymosum, which exerted a hypoglycaemic effect in hyperglycaemic STZ-NA rats. It has a significant antihyperglycemic effect in the pyruvate tolerance test, and it was able to reduce the postprandial hyperglycaemia in maltose tolerance tests significantly. Moreover, it effectively reduced the activity of both gluconeogenic enzymes reaching almost 100% of inhibition, while it presented a modest 32% inhibition of aglucosidases. On the other hand, the extract decreased insulin levels after its oral administration in healthy rats in both nutritional states, without affecting normoglycemia in normal curves and reducing the postprandial peak in glucose load curves. CONCLUSIONS: The traditional consumed form of aerial parts of E. cymosum is safe and regulated glucose levels both in fasting and in postprandial state.

Molecules Isolated from Mexican Hypoglycemic Plants: A Review.

Like in many developing countries, in Mexico, the use of medicinal plants is a common practice. Based on our own field experience, there are at least 800 plants used for treating diabetes nowadays. Thus, their investigation is essential. In this context, this work aims to provide a comprehensive and critical review of the molecules isolated from Mexican hypoglycemic plants, including their source and target tested. In the last few years, some researchers have focused on the study of Mexican hypoglycemic plants. Most works describe the hypoglycemic effect or the mechanism of action of the whole extract, as well as the phytochemical profile of the tested extract. Herein, we analyzed 85 studies encompassing 40 hypoglycemic plants and 86 active compounds belonging to different classes of natural products: 28 flavonoids, 25 aromatic compounds, other than flavonoids, four steroids, 23 terpenoids, 4 oligosaccharides, and 1 polyalcohol. These compounds have shown to inhibit α-glucosidases, increase insulin secretion levels, increase insulin sensitivity, and block hepatic glucose output. Almost half of these molecules are not common metabolites, with a narrow taxonomic distribution, which makes them more interesting as lead molecules. Altogether, this analysis provides a necessary inventory useful for future testing of these active molecules against different hypoglycemic targets, to get a better insight into the already described mechanisms, and overall, to contribute to the knowledge of Mexican medicinal plants.

Glucose-6-Phosphatase and α-Glucosidase Inhibitors from Smilax moranensis Roots Identified by Affinity-Directed Fractionation.

The ethanol/water-soluble extracts of the roots of Smilax moranensis M. Martens & Galeotti, Smilacaceae, which have been appreciated since pre-Hispanic times and traditionally used to treat type 2 diabetes in Mexico, were fractionated by the application of the affinity-directed method to identify glucose-6-phosphatase and α-glucosidase inhibitors. Mass spectrometry was used to identify the inhibitor as free ligands after released from the enzymatic complex by denaturing acidic conditions. The affinity study led to the identification of chlorogenic acid as a glucose-6-phosphatase inhibitor, which is the most abundant metabolite present in the analyzed crude drug. In addition, the affinity studies led to the identification of a flavonolignan glycoside as an α-glucosidase inhibitor. In silico analysis with α-glucosidase MAL12 from the yeast Saccharomyces cerevisiae allowed to determine the best conformational rearrangement for the α-glucosidase inhibitors. Graphical abstract. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s43450-020-00116-3.

Phytochemical Composition and Chronic Hypoglycemic Effect of Bromelia karatas on STZ-NA-Induced Diabetic Rats.

Oral administration of an aqueous extract of the aerial parts of Bromelia karatas to STZ-NA rats showed a significant hypoglycemic effect in a chronic trial lasting 42 days. Chromatographic profiles of the active extract (WE) and an organic extract (OE) of B. karatas were obtained by high-performance liquid chromatography (HPLC) and used to identify their major components. Isolation and identification of the compounds present in the extracts were accomplished by means of various conventional chromatographic and spectroscopic techniques. This process led to the identification of ß-sitosterol-3-O-ß-D-glucopyranoside (1) and ρ-coumaric acid (3) as the major compounds present in the extracts. During the isolation of 1 and 3, seven additional metabolites not previously reported for the plant were obtained, namely, cirsiliol 4'-O-ß-D-glucopyranoside (2), stigmasterol (4), ß-sitosterol (5), 1-O-feruloyl-3-O-ρ-coumaroylglycerol (6), ß-D-(1-O-acetyl-3,6-O-trans-diferuloyl) fructofuranosyl-α-D-2',4',6'-O-triacetylglucopyranoside (7), 1-O-p-coumaroyl-3-O-caffeoylglycerol (8), and 2-propyl-ß-glucopyranoside (9).

Chronic hypoglycemic effect and phytochemical composition of Smilax moranensis roots

ABSTRACT Smilax moranensis M.Martens & Galeotti, Smilacaceae, root is a medicinal plant used among the Chatinos in Oaxaca, Mexico, to control type 2 diabetes. The objectives of the study were to isolate the bioactive compounds from the roots of Smilax and evaluate the chronic hypoglycemic effect of the ethanol-water extract. The main compounds were isolated from the methanolic extract via conventional phytochemical methods. The dried roots of S. moranensis were extracted with methanol and chromatographed on Sephadex LH 20. Fractions were chromatographed and purified on a silica gel chromatography column. The ethanol-water extract was orally administered to hyperglycemic rats for a period of 42 days, and glucose, glycated hemoglobin (HbA1c), and triacylglycerides were measured. Moreover, very-low-density lipoprotein was calculated. During the chemical investigation, three compounds were isolated and characterized, namely, 3-O-caffeoyl-quinic acid, 5-O-caffeoyl-quinic acid and trans-resveratrol, using various spectroscopic techniques. Animal experiments confirmed that the plant extract could control both the glucose and HbA1c levels. In conclusion, this study confirms that the roots of S. moranensis have hypoglycemic properties and suggests that the isolated compounds are potentially involved in this effect.

Morning glory resin glycosides as α-glucosidase inhibitors: In vitro and in silico analysis.

Twenty-seven individual resin glycosides from the morning glory family (Convolvulaceae) were evaluated for their α-glucosidase inhibitory potential. Four of these compounds displayed an inhibitory activity comparable to acarbose, which was used as a positive control. Molecular modeling studies performed by docking analysis were accomplished to predict that the active compounds and acarbose bind to the α-1,4-glucosidase enzyme catalytic site of MAL12 from the yeast Saccharomyces cerevisiae through stable hydrogen bonds primarily with the amino acid residues HIS279 and GLN322. Docking studies with the human maltase-glucoamylase (MGAM) also identified binding modes for resin glycosides inside the catalytic site in the proximity of TYR1251. These results postulate that resin glycosides may be a source of phytotherapeutic agents with antihyperglycemic properties for the prophylaxis and treatment of non-insulin-dependent type 2 diabetes mellitus.

Phytochemical composition and chronic hypoglycemic effect of Rhizophora mangle cortex on STZ-NA-induced diabetic rats

ABSTRACT Type 2 diabetes is a major health problem in Mexico, as it is in other countries, is a chronic condition that develops when the body cannot produce enough insulin or cannot use it appropriately. Both insulin deficiency and insulin resistance lead to high blood glucose levels. In Mexico, people with diabetes are known to use the decoction of red mangrove (Rhizophora mangle L., Rhizophoraceae) bark to control blood glucose levels. Therefore, in this study, we sought to investigate the chronic hypoglycemic and hypolipidemic effects of R. mangle; we also elucidate some of the major phytochemical compounds of R. mangle. To analyze the hypoglycemic and hypolipidemic effects, we used rats with streptozotocin-nicotinamide-induced hyperglycemia; the rats were classified into four groups (six rats each), based on the treatment given, as follows: group 1, non-hyperglycemic control; group 2, hyperglycemic control; group 3, glibenclamide (5 mg/kg body weight); and group 4, Rhizophora ethanol-water extract (90 mg/kg). The extract or glibenclamide was orally administered, dissolved in 1.5 ml of physiological NaCl-solution, twice a day (in the morning and in the evening) over a period of 42 days. The methanolic extract was used to elucidate the main compounds present in R. mangle via conventional phytochemical methods, such as TLC, HLPC, UPLC-DAD-MS, and NMR. The following compounds were detected: cinchonains Ia and Ib, catechin-3-O-rhamnopyranoside, epicatechin, lyoniside, and nudiposide. The daily administration of Rhizophora ethanol-water extract, similar to the traditional usage to control type 2 diabetes, was shown to exert chronic hypoglycemic and hypolipidemic effects. This effect may be associated whit the constituents in the extract. These findings suggest that R. mangle and its constituents could be potentially used to treat type 2 diabetes.

Resistance-modifying Activity in Vinblastine-resistant Human Breast Cancer Cells by Oligosaccharides Obtained from Mucilage of Chia Seeds (Salvia hispanica).

The multidrug resistance (MDR) phenotype is considered as a major cause of the failure in cancer chemotherapy. The acquisition of MDR is usually mediated by the overexpression of drug efflux pumps of a P-glycoprotein. The development of compounds that mitigate the MDR phenotype by modulating the activity of these transport proteins is an important yet elusive target. Here, we screened the saponification and enzymatic degradation products from Salvia hispanica seed's mucilage to discover modulating compounds of the acquired resistance to chemotherapeutic in breast cancer cells. Preparative-scale recycling HPLC was used to purify the hydrolysis degradation products. All compounds were tested in eight different cancer cell lines and Vero cells. All compounds were noncytotoxic at the concentration tested against the drug-sensitive and multidrug-resistant cells (IC50  > 29.2 µM). For the all products, a moderate vinblastine-enhancing activity from 4.55-fold to 6.82-fold was observed. That could be significant from a therapeutic perspective. Copyright © 2017 John Wiley & Sons, Ltd.

Anti-Hyperglycemic Activity of Major Compounds from Calea ternifolia.

Demethylisoencecalin (1) and caleins A (4) and C (5) (3.16-31.6 mg/kg, p.o.), the major components from an infusion of Calea ternifolia controlled postprandial glucose levels during an oral sucrose tolerance test (OSTT, 3 g/kg) in normal and nicotinamide/streptozotocin (NA/STZ, 40/100 mg/kg) hyperglicemic mice. The effects were comparable to those of acarbose (5 mg/kg). During the isolation of 1, 4, and 5, four additional metabolites not previously reported for the plant, were obtained, namely 6-acetyl-5-hydroxy-2-methyl-2-hydroxymethyl-2H-chromene (3), herniarin (6), scoparone (7), and 4',7-dimethylapigenin (8). In addition, the structure of calein C (5) was confirmed by X-ray analysis. Pharmacological evaluation of the essential oil of the species (31.6-316.2 mg/kg, p.o.) provoked also an important decrement of blood glucose levels during an OSTT. Gas chromatography coupled with mass spectrometry (GC-MS) analysis of the headspace solid phase microextraction (HS-SPME)-adsorbed compounds and active essential oil obtained by hydrodistillation revealed that chromene 1 was the major component (19.92%); sesquiterpenes represented the highest percentage of the essential oil content (55.67%) and included curcumene (7.10%), spathulenol (12.95%) and caryophyllene oxide (13.0%). A suitable High Performance Liquid Chromatography (HPLC) method for quantifying chromenes 1 and 6-hydroxyacetyl-5-hydroxy-2,2-dimethyl-2H-chromene (2) was developed and validated according to standard protocols.

Mexican antidiabetic herbs: valuable sources of inhibitors of α-glucosidases.

Type II-diabetes mellitus (TII-DM) has been regarded as one of the most important public health problems in all nations in the 21st century. Although allopathic therapies remain the most important for the initial management of TII-DM, herbal remedies have gained wide acceptance for treating this condition. These alternative therapies are particularly valued in countries such as Mexico, rich in medicinal plants strongly attached to the cultural values of the population. Medicinal plants are prized sources of α-glucosidase inhibitors, which delay the liberation of glucose from complex carbohydrates, retarding glucose absorption, and thus controlling the characteristic hyperglycemia of TII-DM. Among the plant species used for treating diabetes in Mexico only 38 have been analyzed for their inhibitory activity of α-glucosidases. Most of these studies, reviewed in the present work, have focused on the evaluation of different types of extracts on the activity of α-glucosidases from diverse sources. Four species have been thoroughly analyzed in order to discover novel α-glucosidase inhibitors, namely, Hintonia latiflora and Hintonia standleyana (Rubiaceae), Ligusticum porteri (Apiaceae), and Brickellia cavanillesii (Asteraceae). Their ethnomedical uses, pharmacological and toxicological studies, chemical composition, and antihyperglycemic principles with α-glucosidase inhibitory activity are summarized.

α-glucosidase inhibitors from Brickellia cavanillesii.

An aqueous extract from the aerial parts of Brickellia cavanillesii attenuated postprandial hyperglycemia in diabetic mice during oral glucose and sucrose tolerance tests. Experimental type-II DM was achieved by treating mice with streptozotocin (100 mg/kg) and ß-nicotinamide adenine dinucleotide (40 mg/kg). These pharmacological results demonstrated that B. cavanillesii is effective for controlling fasting and postprandial blood glucose levels in animal models. The same aqueous extract also showed potent inhibitory activity (IC(50) = 0.169 vs 1.12 mg/mL for acarbose) against yeast α-glucosidase. Bioassay-guided fractionation of the active extract using the α-glucosidase inhibitory assay led to the isolation of several compounds including two chromenes [6-acetyl-5-hydroxy-2,2-dimethyl-2H-chromene (1) and 6-hydroxyacetyl-5-hydroxy-2,2-dimethyl-2H-chromene (2)], two sesquiterpene lactones [caleins B (3) and C (4)], several flavonoids [acacetin (5), genkwanin (6), isorhamnetin (7), kaempferol (8), and quercetin (9)], and 3,5-di-O-caffeoylquinic acid (10). Chromene 2 is a new chemical entity. Compounds 2, 4, 7, and 9 inhibited the activity of yeast α-glucosidase with IC(50) 0.42, 0.28, 0.16, and 0.53 mM, respectively, vs 1.7 mM for acarbose. Kinetic analysis revealed that compounds 4 and 7 behaved as mixed-type inhibitors with K(i) values of 1.91 and 0.41 mM, respectively, while 2 was noncompetititive, with a K(i) of 0.13 mM. Docking analysis predicted that these compounds, except 2, bind to the enzyme at the catalytic site.