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Background: MicroRNAs expressions that are known as Posttranscriptional repressors of gene expression in animals and plants have an important role in diabetic consequences and pathogenesis. This research aimed to investigate the expression of miR-34a, miR-126, and miR-125a-5p levels for diagnosis of diabetes, and the effect of α-L-guluronic acidand β-D-mannuronic acid supplementation on expression patterns of these MicroRNAs in diabetic rats.Methods: Total MicroRNAs levels were measured by Quantitative reverse transcription polymerase chain reaction (qRT-PCR) method. Body weight, fasting insulin level, food intake and Vitamin D level in four groups were also assessed after 4 weeks.Results: The analyses illustrate marked declines in expression of miR-126 (p=0.009) and miR-125a-5p (p=0.019) levels in the control diabetic group when compared to the control healthy group. Expression levels of miR-126 (p=0.026) and miR-125a-5p (p=0.041) were upregulated considerably in diabetic rats after treatment with β-D-mannuronic acid and α-L-guluronic acid, respectively.Conclusion: These results suggest that use of miR-126 and 125a-5p is a sensible criterion for diagnosing diabetes. β-D-mannuronic acid is also considered a new therapeutic agent to correct disturbances in the MicroRNAs profile of diabetic rats.

Background & objectives: To examine β-D-mannuronic acid (M2000) effects on L-selectin shedding and leucocyte function-associated antigen-1 (LFA-1) expression as mechanisms of action of this drug in patients with ankylosing spondylitis (AS). Methods: To investigate the molecular consequences of β-D-mannuronic acid on L-selectin shedding, flow cytometry method was used. Furthermore, the effect of it on LFA-1 gene expression was analyzed by using quantitative real time (qRT)-PCR technique. Results: The LFA-1 expression in patients with AS was higher than controls (P=0.046). The LFA-1 expression after 12 wk therapy with β-D-mannuronic acid was meaningfully decreased (P=0.01). After 12 wk treatment with β-D-mannuronic acid, the frequency of CD62L-expressing CD4+ T cells in patients with AS, was not considerably altered, compared to the patients before therapy (P=0.5). Furthermore, after 12 wk therapy with β-D-mannuronic acid, L-selectin expression levels on CD4+ T-cells in patients with AS, were not remarkably changed, compared to the expression levels of these in patients before treatment (P=0.2). Interpretation & conclusions: The results of this study for the first time showed that β-D-mannuronic acid can affect events of adhesion cascade in patients with AS. Moreover, β-D-mannuronic acid presented as an acceptable benefit to AS patients and could aid in the process of disease management. [ABSTRACT FROM AUTHOR]

The discovery of hematologic improvement and bone marrow modification by the drug β‐D mannuronic acid (M2000) during treatment of rheumatoid arthritis in phase 1/2/3 clinical trials prompted us to design a new trial to target hematologic deficits in myelodysplastic syndromes (MDS). In this open‐label, randomized phase 2 clinical trial, the potential effect and tolerability of drug M2000 was assessed in patients with low‐ and intermediate‐1‐risk MDS. The primary efficacy end point was hematologic improvement after 12 weeks of β‐D‐mannuronic acid therapy. Among 34 enrolled patients, half received their conventional therapy plus β‐D‐mannuronic acid, and the other half received only conventional drugs. In the conventional + β‐D mannuronic acid treatment group, hematologic improvement and development of transfusion independence and/or reduction in transfusion requirements were seen in 12 patients (92.3%) and 1 patient (7.7%), respectively. Moreover, 5 patients (38.5%), 2 patients (15.4%), and 1 patient (7.7%) in the β‐D‐mannuronic acid‐treated group showed hematologic improvement of the major parameters of erythroid, neutrophil, and platelet responses, respectively, based on the International Working Group criteria), whereas in the conventional treatment group as control, no hematologic improvements including erythroid, neutrophil, and platelet response was seen. In this trial, the addition of β‐D mannuronic acid to conventional treatment showed promising results in MDS patients with low and intermediate‐1 risk with effects on hematologic improvements without significant adverse effect. [ABSTRACT FROM AUTHOR]

Context: miR-146a, its targets (IRAK1, TRAF6) and NF-κB transcription factor play a fundamental role in rheumatoid arthritis (RA). Positive effects of drug β-d-mannuronic acid (M2000) were proven on their expression in the HEK-Blue hTLR2 cell line, and results of its phase III clinical trial on RA patients were encouraging. Objective: This research aimed to investigate the effects of M2000 on expression of these genes and serum levels of IL-6 and TNF-α as pro-inflammatory cytokines in RA patients. Material and methods: In this study (Trial Registration Number: IRCT2017100213739N10), 12 RA patients (according to the American College of Rheumatology criteria) and 12 healthy subjects (as control group) were selected. The gene expression of miR-146a, IRAK1, TRAF6, and NF-κB were measured at the baseline and after 12 weeks M2000 therapy, using quantitative real-time PCR method. Moreover, the serum levels of IL-6 and TNF-α were evaluated at the similar times by ELISA method. Results: Our findings showed that the gene expression of miR-146a, IRAK1, TRAF6, and NF-κB significantly decreased after 12 weeks M2000 therapy in RA patients (0.81-, 0.68-, 0.79-, 0.82-fold, with p <.05, p <.01, p <.01, p <.05, respectively). Furthermore, the serum levels of IL-6 and TNF-α significantly reduced in these patients after 12 weeks M2000 therapy (both with p <.05). Conclusions: The present research results determined the part of molecular mechanisms of drug M2000 in RA treatment, based on the expression and function modification of miR-146a, IRAK1, TRAF6, NF-κB, IL-6 and TNF-α. [ABSTRACT FROM AUTHOR]

Summary: Following the potent efficacy of β‐D‐Mannuronic acid in a breast cancer murine model, we evaluated the efficacy of this novel non‐steroidal anti‐inflammatory drug in breast cancer patients in the present clinical trial. The study was an 8‐week randomized, controlled, phase II clinical trial (IRCT: 2017012213739N7 (in 48 pre‐surgical breast cancer patients. Patients who had breast cancer at early stage, with invasive ductal carcinoma, were placed on a waiting‐list for surgery and were allocated to the study. β‐D‐Mannuronic was administrated at a dose of two capsules (1000 mg/d) orally during a period of 8 weeks. The end point of this study was when the patients were admitted for surgery. Moreover, the patients' well‐being status was followed up on for safety. There were no statistically significant differences between treatment and non‐treatment groups at baseline. β‐D‐Mannuronic acid therapy, from 20 patients, showed that in one patient (5%) tumour size was decreased; in five patients (25%) tumour growth was stopped; and in 14 patients (70%) the growth rate in the treatment group did not show significant change, compared to the non‐treatment group. Evaluation of two tumour markers (carcinoembryonic antigen and cancer antigen 15‐3) showed that there was no significant difference between before and after treatment. Although the use of some non‐steroidal anti‐inflammatory drugs in a long time period has shown a prophylactic effect in breast cancer, their therapeutic efficacy in a short time period is unknown, whereas treatment with β‐D‐Mannuronic acid during 8 weeks could show 30% therapeutic effects in pre‐surgical breast cancer patients. [ABSTRACT FROM AUTHOR]

With respect to the role of chronic inflammation in the induction and progression of breast cancer (BC). The relationship between tumor and tumor microenvironment may be a hopeful strategy for BC therapy. According to the effect of β-D-Mannuronic acid (M2000) as a novel non-steroidal anti-inflammatory drug (NSAID) on BC murine model and 4T1 cell line, we started to study that was a phase II, randomized, controlled clinical trial. 24 women with BC were included in this study and were followed by fixed oral doses of M2000, 500 mg two times a day (6-8 weeks). Blood samples were collected at baseline and weeks 6-8. To compare the patterns of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), C-C motif chemokine ligand 22 (CCL22) and The transforming growth factor-beta 1 (TGFβ1) gene expression and T regulatory cells (Tregs) frequency of healthy women normal controls with BC patients, a set of 10 blood samples of women healthy volunteers was collected. The gene expression was evaluated by quantitative Real-time PCR (qRT-PCR) and the frequency of Tregs was assessed by flow cytometry. Our results showed, reduction in MMP-2 (p=0.08), MMP-9 (p=0.03), CCL22 (p=0.003) and TGFβ1 (p=0.1) gene expression and Tregs frequency (p=0.01) which play a main role in the development of chronic inflammation, angiogenesis, tumorigenesis and metastasis. Our findings demonstrated that M2000 therapy as a novel designed NSAID had valuable therapeutic effects on BC. No adverse effects were observed following the use of M2000 after 6-8 weeks.

Metastasis is the main cause of death in breast cancer patients. Inflammatory processes following crosstalk between tumor cells and tumor microenvironment play an important role in progression and metastasis of cancer. Hence, targeting of these interactions may represent a novel promising strategy for breast cancer therapy. So, we investigated the effects of β-D mannuronic acid (BDM), a new antiinflammatory agent, on 4T1 breast cancer cell line both in vitro and in vivo. Proliferation assays revealed low-cytotoxic effect of BDM on 4T1 cells. However, BDM reduced activity of MMP-2, MMP-9 and significantly decreased the adhesion of 4T1 cells to extracellular matrix (ECM) in a dose-dependent manner. The in vivo results demonstrated that BDM strongly inhibits tumor growth and increases lifespan as compared with control mice. The decrease in tumor mass was associated with decreased metastasis, recruitment, and frequency of inflammatory cells in tumor tissue. Our preclinical findings demonstrated that BDM therapy not only prevents formation of chronic inflammatory response but also inhibits crosstalk between tumor cells and their microenvironment, which is associated with reduction of tumor growth and metastasis arrest. Our data imply the use of BDM therapy in future clinical trials to open a new horizon for breast cancer therapy. [ABSTRACT FROM AUTHOR]

Context: β-d-Mannuronic acid (M2000) has shown its therapeutic effects with the greatest tolerability and efficacy in various experimental models such as experimental autoimmune encephalomyelitis (EAE), adjuvant induced arthritis (AIA), nephrotic syndrome, and acute glomerulonephritis. Despite pharmacological effects of β-D-mannuronic acid, there have been no systematic toxicological studies on its safety so far. Objective: The study was designed to determine the acute and subchronic toxicity of β-D-mannuronic acid, an anti-inflammatory agent, in healthy male NMRI mice and Wistar rats, respectively. Materials and methods: For the acute toxicity study, the animals received orally five different single doses of β-D-mannuronic acid and were kept under observation for 14 d. In the subchronic study, 24 Wistar male rats were divided into four groups and were treated orally (gavage) once daily with test substance preparation at dose levels of 0, 50, 250, and 1250 mg/kg body weight for at least 63 consecutive days (9 weeks). Mortality, clinical signs, body weight changes, hematological and biochemical parameters, gross findings, organ weights, and histopathological determinations were monitored during the study. Results: The results of acute toxicity indicated that the LD50of β-D-mannuronic acid is 4.6 g/kg. We found no mortality and no abnormality in clinical signs, body weight, relative organ weights, or necropsy in any of the animals in the subchronic study. Additionally, the results showed no significant difference in hematological, biochemical, and histopathological parameters in rats. Conclusions: Our results suggest that β-D-mannuronic acid is relatively safe when administered orally in animals. [ABSTRACT FROM PUBLISHER]

The dominant impact of the α-L-guluronic acid chain on regulation of the mass transfer character of calcium alginate membranes was investigated. The polymer frameworks of the membranes were successfully regulated by altering the mass fraction of homopolymeric blocks of α-L-guluronic acid (FGG) in the entire molecular chain of alginate. Sodium alginate is well-known as a hydrophilic polysaccharide. Its molecular chain is composed of α-L-guluronic acid and β-D-mannuronic acid. The polymeric structure of calcium alginate is mainly constructed by intermolecular ionic bonds with homopolymeric blocks of the α-L-guluronic acid junction zone. The entire alginate polymer chain then forms into a swollen gel and a membrane. The water fraction of the swollen membrane and the mechanical strength changed with FGG. The volumetric water fraction based on the swollen state of the membrane was evaluated from the gravitational water content (HM). The change in the mass transfer mechanism of the membrane was evident in the effective diffusion coefficient, especially for the smaller molecules (e.g., urea). Good correlation between the volumetric water fraction and the effective diffusion coefficient strongly suggested that the mass transfer channels in the alginate membrane were dominantly regulated by the mass fraction of homopolymeric blocks of α-L-guluronic acid (FGG). The relationship between the effective diffusion coefficient and the volumetric water fraction agreed with Yasuda's free volume theory. The tortuosity was markedly increased and the effective diffusion coefficient was reduced. In conclusion, the homopolymeric blocks of α-L-guluronic acid have a dominant impact on the mass transfer character in the calcium alginate membrane. [ABSTRACT FROM AUTHOR]

Objectives: The goal of this article is to retrace the studies of β-D-Mannuronic Acid (M2000) as a new immunosuppressive drug with non-steroidal anti-inflammatory drugs (NSAIDs) property in miscellaneous aspects including in vitro, in vivo examinations, clinical trials and related to clinical trials studies. Our goal is to compare the effect of this drug with other similar drugs through varied researches and to follow tolerability, biocompatibility, potency, safety, and efficacy of this medication in different studies, as well as to evaluate its therapeutic effectiveness in various diseases., Materials and Methods: Different methods were applied in the studies of β-D-Mannuronic Acid under in vitro, in vivo examinations, and clinical trials phase I, II and III and related investigations to these clinical trials using different techniques showing the efficacy of this medication in the treatment of various diseases., Results: The administration of β -D-Mannuronic Acid showed the greatest tolerability and biocompatibility compared to diclofenac, piroxicam, and dexamethasone without or very low side effects. The drug has shown a punchy effect on many molecules which participate either in physiologic or in pathogenic activities in animal models and human. This new drug not only revealed the anti-inflammatory and immunosuppressive properties but also based on the results of various investigations, β-D-Mannuronic Acid showed the antidiabetic, cardioprotective and anti-tumoral effects., Conclusion: β-D-Mannuronic Acid (M2000) as a novel immunosuppressive drug with NSAID properties along with antidiabetic, cardioprotective and anti-tumoral efficacy showed great tolerability and safety profile. In addition, it has no or mild adverse events compared with many other medicines, therefore this medicament could be considered as a landmark in pharmacology and represent turn point in the treatment of different diseases based on the experimental and in vitro studies explained and clinical and related studies proved., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Based on the encouraging results of phase III clinical trial of β-Dmannuronic acid (M2000) (as a new anti-inflammatory drug) in patients with RA, in this study, we aimed to evaluate the effects of this drug on the expression of chemokines and their receptors in PBMCs of RA patients., Methods: PBMCs of RA patients and healthy controls were separated and the patients' cells were treated with low, moderate and high doses (5, 25 and 50 μg/mL) of M2000 and optimum dose (1 μg/mL) of diclofenac, as a control in RPMI-1640 medium. Real-time PCR was used for evaluating the mRNA expression of CXCR3, CXCR4, CCR2, CCR5 and CCL2/MCP-1. Cell surface expression of CCR2 was investigated using flow cytometry., Results: CCR5 mRNA expression reduced significantly, after treatment of the patients' cells with all three doses of M2000 and optimum dose of diclofenac. CXCR3 mRNA expression was downregulated significantly followed by the treatment of these cells with moderate and high doses of M2000 and optimum dose of diclofenac. CXCR4 mRNA expression declined significantly after the treatment of these cells with moderate and high doses of M2000. CCL2 mRNA expression significantly reduced only followed by the treatment of these cells with a high dose of M2000, whereas, mRNA and cell surface expressions of CCR2 diminished significantly followed by the treatment of these cells with a high dose of M2000 and optimum dose of diclofenac., Conclusion: According to our results, M2000 through the down-regulation of chemokines and their receptors may restrict the infiltration of immune cells into the synovium., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

With respect to the role of chronic inflammation in the induction and progression of breast cancer (BC). The relationship between tumor and tumor microenvironment may be a hopeful strategy for BC therapy. According to the effect of β-D-Mannuronic acid (M2000) as a novel non-steroidal anti-inflammatory drug (NSAID) on BC murine model and 4T1 cell line, we started to study that was a phase II, randomized, controlled clinical trial. 24 women with BC were included in this study and were followed by fixed oral doses of M2000, 500 mg two times a day (6-8 weeks). Blood samples were collected at baseline and weeks 6-8. To compare the patterns of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), C-C motif chemokine ligand 22 (CCL22) and The transforming growth factor-beta 1 (TGFβ1) gene expression and T regulatory cells (Tregs) frequency of healthy women normal controls with BC patients, a set of 10 blood samples of  women healthy volunteers was collected. The gene expression was evaluated by quantitative Real-time PCR (qRT-PCR) and the frequency of Tregs was assessed by flow cytometry. Our results showed, reduction in MMP-2 (p=0.08), MMP-9 (p=0.03), CCL22 (p=0.003) and TGFβ1 (p=0.1) gene expression and Tregs frequency (p=0.01) which play a main role in the development of chronic inflammation, angiogenesis, tumorigenesis and metastasis. Our findings demonstrated that M2000 therapy as a novel designed NSAID had valuable therapeutic effects on BC. No adverse effects were observed following the use of M2000 after 6-8 weeks.

Objective: To evaluate the efficacy, safety and tolerability of β-d-mannuronic acid (M2000) in the treatment of ankylosing spondylitis (AS)., Methods: The study was a 12-week randomized, double-blind, placebo-controlled, phase I/II clinical trial with 3 treatment arms: placebo, β-d-mannuronic acid and naproxen. Patients who had AS according to the modified New York criteria, with active disease at baseline were eligible for study. Primary outcome measure was the Assessment of SpondyloArthritis international Society (ASAS) 20 response rate at week 12., Results: Of the 85 randomized patients, 27 were allocated to receive placebo, 28 naproxen, and 30 β-d-mannuronic acid. There were no statistically significant differences between treatment groups at baseline. Of the patients receiving β-d-mannuronic acid, 57.7% achieved an ASAS20 response at week 12, compared with 59% of the patients in the naproxen group (P>0.05) and 19% of the patients in the placebo group (P=0.007). In comparison with patients receiving placebo over the 12-week treatment period, those receiving β-d-mannuronic acid and naproxen demonstrated statistically significantly greater improvement in all secondary endpoints. Interestingly, β-d-mannuronic acid reduced some parameters associated with inflammation more effectively than naproxen and placebo. The incidence of gastrointestinal and other adverse events were higher on naproxen than on β-d-mannuronic acid and placebo., Conclusion: The present study demonstrated similar efficacy, but with a more favorable safety profile for β-d-mannuronic acid than naproxen and, therefore, suggest that β-d-mannuronic acid is suitable for the management of AS., Trial Registration: Iranian registry of clinical trials; www.irct.ir; IRCT2013062213739N1., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: The aim of this study was to evaluate the effect of ß-Dmannuronic acid (M2000) on related miRNAs to dendritic cells (DCs) differentiation. DC-based immunosuppressive drugs can suppress the progression of autoimmune diseases, however, their notable side effects in increasing the risk of infectious diseases and cancers should be considered. The β-D-mannuronic acid, as a novel non-steroidal anti-inflammatory agent, has been tested in various experimental models., Method: The effect of M2000 on expression of miRNA-155 and miRNA-221 was examined. To investigate how M2000 affects differentiation of human dendritic DCs in a defined inflammatory environment, human peripheral blood mononuclear cells were isolated from healthy blood and the monocytes were purified using anti-CD14 microbeads. The so isolated monocytes were subsequently incubated in the presence of M2000 in two different doses (3 and 6 mMol/well) adding granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 for inducing monocytes to immature DC and lipopolysaccharide for running DC differentiation. The expression of miRNA-155 and miRNA-221 were examined with Real Time PCR., Results: The results demonstrate that M2000 has no significant side effect on expression of miR-155 and miR-221 in both immature DC and mature DC process in vitro., Conclusion: Our findings show that β-D-mannuronic acid is a safe agent which has no adverse effect on regulatory miRNA-155 and miRNA-221 in dendritic cells., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)