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1.
J Agric Food Chem ; 67(32): 9070-9078, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31343168

RESUMO

In this study, an immunologically active novel microparticulate mushroom ß-glucan (PRA-1p) was prepared using an alkali-soluble glucan PRA-1 by an emulsification and cross-linking method. PRA-1 was a hyperbranched (1→3),(1→6)-ß-d-glucan with a degree of branching of 0.89, isolated from the sclerotia of Polyporus rhinocerus. PRA-1 had a rod-like conformation, while PRA-1p exhibited a monodisperse and homogeneous spherical conformation with a diameter ranging from 0.3 to 2.0 µm in water. PRA-1p significantly induced nitric oxide and reactive oxygen species production as well as morphological changes of murine macrophages (RAW 264.7 cells) and upregulated their phagocytic activity. Furthermore, PRA-1p treatment markedly enhanced the secretion of cytokines, including cutaneous T cell-attracting chemokine 27, granulocyte-colony-stimulating factor, monocyte chemoattractant protein 1, macrophage inflammatory protein 1α, macrophage inflammatory protein 2, regulated on activation, normal T cell expressed and secreted, soluble tumor necrosis factor receptor 1, and tissue inhibitors of metalloproteinases. Activation of RAW 264.7 cells triggered by PRA-1p was associated with activation of inducible nitric oxide synthase, nuclear factor κB, extracellular signal-regulated kinase, and protein kinase B. This work suggests that novel PRA-1p derived from the mushroom sclerotia of P. rhinocerus has potential application as an immunostimulatory agent.


Assuntos
Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polyporus/química , beta-Glucanas/química , beta-Glucanas/farmacologia , Animais , Quimiocina CCL27/genética , Quimiocina CCL27/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Fatores Imunológicos/isolamento & purificação , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , NF-kappa B/genética , NF-kappa B/imunologia , Óxido Nítrico/imunologia , Fagocitose/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Polyporus/imunologia , Células RAW 264.7 , Espécies Reativas de Oxigênio/imunologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , beta-Glucanas/isolamento & purificação
2.
Eur J Med Chem ; 178: 13-29, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31173968

RESUMO

The oncogenic Epstein-Barr virus (EBV) evades the immune system through limiting the expression of its highly antigenic and essential genome maintenance protein, EBNA1, to the minimal level to ensure viral genome replication, thereby also minimizing the production of EBNA1-derived antigenic peptides. This regulation is based on inhibition of translation of the virally-encoded EBNA1 mRNA, and involves the interaction of host protein nucleolin (NCL) with G-quadruplex (G4) structures that form in the glycine-alanine repeat (GAr)-encoding sequence of the EBNA1 mRNA. Ligands that bind to these G4-RNA can prevent their interaction with NCL, leading to disinhibition of EBNA1 expression and antigen presentation, thereby interfering with the immune evasion of EBNA1 and therefore of EBV (M.J. Lista et al., Nature Commun., 2017, 8, 16043). In this work, we synthesized and studied a series of 20 cationic bis(acylhydrazone) derivatives designed as G4 ligands. The in vitro evaluation showed that most derivatives based on central pyridine (Py), naphthyridine (Naph) or phenanthroline (Phen) units were efficient G4 binders, in contrast to their pyrimidine (Pym) counterparts, which were poor G4 binders due to a significantly different molecular geometry. The influence of lateral heterocyclic units (N-substituted pyridinium or quinolinium residues) on G4-binding properties was also investigated. Two novel compounds, namely PyDH2 and PhenDH2, used at a 5 µM concentration, were able to significantly enhance EBNA1 expression in H1299 cells in a GAr-dependent manner, while being significantly less toxic than the prototype drug PhenDC3 (GI50 > 50 µM). Antigen presentation, RNA pull-down and proximity ligation assays confirmed that the effect of both drugs was related to the disruption of NCL-EBNA1 mRNA interaction and the subsequent promotion of GAr-restricted antigen presentation. Our work provides a novel modular scaffold for the development of G-quadruplex-targeting drugs acting through interference with G4-protein interaction.


Assuntos
Hidrazonas/farmacologia , Evasão da Resposta Imune/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Fosfoproteínas/metabolismo , Ligação Proteica/efeitos dos fármacos , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/metabolismo , Animais , Linhagem Celular Tumoral , Antígenos Nucleares do Vírus Epstein-Barr/genética , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Quadruplex G , Herpesvirus Humano 4/genética , Humanos , Hidrazonas/síntese química , Hidrazonas/química , Fatores Imunológicos/síntese química , Fatores Imunológicos/química , Ligantes , Camundongos , RNA Mensageiro/genética
3.
Eur J Med Chem ; 178: 243-258, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31185414

RESUMO

To address the multifactorial nature of Alzheimer's Disease (AD), a multi-target-directed ligand approach was herein developed. As a follow-up of our previous studies, a small library of newly designed 2-arylbenzofuran derivatives was evaluated towards cholinesterases and cannabinoid receptors. The two most promising compounds, 8 and 10, were then assessed for their neuroprotective activity and for their ability to modulate the microglial phenotype. Compound 8 emerged as able to fight AD from several directions: it restored the cholinergic system by inhibiting butyrylcholinesterase, showed neuroprotective activity against Aß1-42 oligomers, was a potent and selective CB2 ligand and had immunomodulatory effects, switching microglia from the pro-inflammatory M1 to the neuroprotective M2 phenotype. Derivative 10 was a potent CB2 inverse agonist with promising immunomodulatory properties and could be considered as a tool for investigating the role of CB2 receptors and for developing potential immunomodulating drugs addressing the endocannabinoid system.


Assuntos
Benzofuranos/farmacologia , Inibidores da Colinesterase/farmacologia , Fatores Imunológicos/farmacologia , Fármacos Neuroprotetores/farmacologia , Bibliotecas de Moléculas Pequenas/farmacologia , Acetilcolinesterase/metabolismo , Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/antagonistas & inibidores , Animais , Benzofuranos/síntese química , Benzofuranos/química , Benzofuranos/metabolismo , Butirilcolinesterase/química , Butirilcolinesterase/metabolismo , Células CHO , Domínio Catalítico , Linhagem Celular Tumoral , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Inibidores da Colinesterase/metabolismo , Cricetulus , Desenho de Drogas , Humanos , Fatores Imunológicos/síntese química , Fatores Imunológicos/química , Fatores Imunológicos/metabolismo , Camundongos , Microglia/efeitos dos fármacos , Simulação de Acoplamento Molecular , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/metabolismo , Fragmentos de Peptídeos/antagonistas & inibidores , Ligação Proteica , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/metabolismo , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/metabolismo
4.
Nat Chem ; 11(6): 521-532, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31086302

RESUMO

The chemical diversification of natural products provides a robust and general method for the creation of stereochemically rich and structurally diverse small molecules. The resulting compounds have physicochemical traits different from those in most screening collections, and as such are an excellent source for biological discovery. Herein, we subject the diterpene natural product pleuromutilin to reaction sequences focused on creating ring system diversity in few synthetic steps. This effort resulted in a collection of compounds with previously unreported ring systems, providing a novel set of structurally diverse and highly complex compounds suitable for screening in a variety of different settings. Biological evaluation identified the novel compound ferroptocide, a small molecule that rapidly and robustly induces ferroptotic death of cancer cells. Target identification efforts and CRISPR knockout studies reveal that ferroptocide is an inhibitor of thioredoxin, a key component of the antioxidant system in the cell. Ferroptocide positively modulates the immune system in a murine model of breast cancer and will be a useful tool to study the utility of pro-ferroptotic agents for treatment of cancer.


Assuntos
Antineoplásicos/farmacologia , Azulenos/farmacologia , Morte Celular/efeitos dos fármacos , Diterpenos/farmacologia , Piridazinas/farmacologia , Tiorredoxinas/antagonistas & inibidores , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Azulenos/síntese química , Azulenos/química , Linhagem Celular Tumoral , Cisteína/química , Diterpenos/síntese química , Diterpenos/química , Humanos , Fatores Imunológicos/síntese química , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Camundongos SCID , Estrutura Molecular , Piridazinas/síntese química , Piridazinas/química , Relação Estrutura-Atividade , Tiorredoxinas/química
5.
Carbohydr Polym ; 216: 270-281, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31047067

RESUMO

The fine structure and chain conformation of a heteropolysaccharide (PCIPS3) from mycelium of Paecilomyces cicadae were investigated via the analysis of HPLC, IR, methylation, NMR spectroscopy and multiangle light scattering. It was determined to be a 2.23 × 104 g/mol heteropolysaccharide primarily composed of glucose, galactose and mannose in a molar ratio of 23.8:2.1:1.0. The PCIPS3 backbone consisted of 1,4-linked α-d-Glcp and 1,4-linked 6-O-Me-α-d-Glcp residues, which were occasionally interrupted by branched ß-Galf residues through 1,6-linkage. Moreover, the α (0.60) from Mark-Houwink-Sakurada (MHS) equation suggested that PCIPS3 adopted a flexible chain conformation in 0.1 mol/L NaNO3 at 25 °C. The worm-like chains model parameters for PCIPS3 were estimated as following: ML = 437 nm-1, q = 0.46 nm and 0.79 nm, which were further evidenced by AFM. Furthermore, PCIPS3 showed excellent scavenging capacities of 2,2-diphenyl-1-picrylhydrazyl radical, superoxide radical, hydroxyl radical, ORAC radical and moderate immunomodulatory activity.


Assuntos
Depuradores de Radicais Livres/farmacologia , Polissacarídeos Fúngicos/farmacologia , Fatores Imunológicos/farmacologia , Paecilomyces/química , Animais , Configuração de Carboidratos , Sobrevivência Celular/efeitos dos fármacos , Depuradores de Radicais Livres/química , Depuradores de Radicais Livres/isolamento & purificação , Depuradores de Radicais Livres/toxicidade , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/isolamento & purificação , Polissacarídeos Fúngicos/toxicidade , Radical Hidroxila/química , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/toxicidade , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Camundongos , Peróxidos/química , Células RAW 264.7 , Superóxidos/química , Fator de Necrose Tumoral alfa/metabolismo
6.
Cell Mol Biol (Noisy-le-grand) ; 65(4): 43-47, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-31078151

RESUMO

The immune system is an important physiological defense system. Its balance and stability are closely related to the body's health. Once the immune system loses its dynamic balance, the immune response will be blocked, which will lead to the occurrence of various diseases. Hesperetin is a kind of natural flavonoids extracted from citrus fruits of Rutaceae and it has many pharmacological activities. However, its water solubility and liposolubility are poor, and it is easy to be quickly metabolized in vivo, so it is difficult to maintain high blood drug concentration. Therefore, its derivative (HES) was found by structural modification. In this study, THP-1 cells were used as experimental model to investigate the immunomodulatory effect of HES in vitro. The results showed that HES participates in immune response by enhancing phagocytosis of macrophages to promote the release of NO, IL-6 and IL-1ß, and enhancing immunity by up-regulating the expression of Bcl-2 and Bcl-XL proteins. This study provides a theoretical and practical basis for the development of HES as an immunomodulator in the future.


Assuntos
Hesperidina/farmacologia , Fatores Imunológicos/farmacologia , Macrófagos/imunologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Hesperidina/química , Humanos , Fatores Imunológicos/química , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Óxido Nítrico/biossíntese , Fagocitose/efeitos dos fármacos , Células THP-1 , Proteína bcl-X/metabolismo
7.
Expert Opin Ther Pat ; 29(5): 339-351, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31064237

RESUMO

INTRODUCTION: Boron-containing compounds induce effects on immune responses. Such effects are interesting to the biomedical field for the development of therapeutic tools to modulate the immune system. AREAS COVERED: The scope of BCC use to modify immune responses is expanding, mainly with regard to inflammatory diseases. The information was organized to demonstrate the breadth of reported effects. BCCs act as modulators of innate and adaptive immunity, with the former including regulation of cluster differentiation and cytokine production. In addition, BCCs exert effects on inflammation induced by infectious and noninfectious agents, and there are also reports regarding their effects on mechanisms involving hypersensitivity and transplants. Finally, the authors discuss the beneficial effects of BCCs on pathologies involving various targets and mechanisms. EXPERT OPINION: Some BCCs are currently used as drugs in humans. The mechanisms by which these BCCs modulate immune responses, as well as the required structure-activity relationship for each observed mechanism of action, should be clarified. The former will allow for the development of improved immunomodulatory drugs with extensive applications in medicine. Patenting trends involve claims concerning the synthesis and actions of identified molecules with a defined profile regarding cytokines, cell differentiation, proliferation, and antibody production.


Assuntos
Compostos de Boro/farmacologia , Fatores Imunológicos/farmacologia , Inflamação/tratamento farmacológico , Imunidade Adaptativa/efeitos dos fármacos , Animais , Formação de Anticorpos/efeitos dos fármacos , Compostos de Boro/química , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocinas/imunologia , Humanos , Imunidade Inata/efeitos dos fármacos , Fatores Imunológicos/química , Inflamação/imunologia , Patentes como Assunto , Relação Estrutura-Atividade
8.
Enzyme Microb Technol ; 127: 50-57, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31088616

RESUMO

Macrophages eliminate and destroy invading bacteria and contaminants by engulfing them or secreting cytokines that trigger downstream immune responses. Consequently, impairment of the phagocytic functions of macrophages and/or suppressing their cytokine secretion are dangerous to organisms that rely on immune protection. Accordingly, exposure to environmental nanoparticles (NPs) that display immunomodulatory properties are serious. In this work, two types of NPs, i.e., mild-toxicity CuInS2 NPs and high-toxicity CdTe NPs, were used to evaluate the effects of NP exposure for macrophages. Following incubation for 24 h, THP-1-derived macrophage viability was assessed using an MTT method after exposing the THP-1 cells to different concentrations of CuInS2 or CdTe NPs. Phagocytosis assays demonstrated that both CuInS2 and CdTe NPs impair phagocytic activity toward Staphylococcus aureus (S. aureus). After pretreatment with CuInS2 and CdTe NPs at 4 µmol/L, THP-1 macrophages exhibited decreases in phagocytic ratio from ca. 32.9% to ca. 18.5% and 18.7%, respectively. Since the zeta potentials of intact and weathered CuInS2 NPs were distributed over a wide range from positive to negative, large quantities of intact and weathered CuInS2 NPs bore sufficient positive charge on their surfaces to induce membrane depolarization, thus theoretically providing electrostatic forces between S. aureus and THP-1, which could induce downstream intracellular events that increase phagocytosis. However, real time polymerase chain reaction arrays revealed that transcription of the pro-inflammatory factors IL-1ß, IL-6, and TNF-α decreased while that of the anti-inflammatory factor IL-10 increased after treatment with CuInS2 NPs. Furthermore, transcription of TNF-α decreased while IL-10 increased after treatment with CdTe NPs. Thus, both kinds of NPs inhibited phagocytosis of S. aureus by THP-1 to some extent, confirming that immunosuppression can occur when macrophages are exposed to environmental NPs.


Assuntos
Citocinas/metabolismo , Fatores Imunológicos/metabolismo , Macrófagos/efeitos dos fármacos , Nanopartículas/metabolismo , Fagocitose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Fatores Imunológicos/química , Macrófagos/metabolismo , Nanopartículas/química , Staphylococcus aureus/imunologia , Células THP-1
9.
Carbohydr Polym ; 217: 79-89, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31079688

RESUMO

Several scientific evidences have revealed probiotics implications on human health. A wide range of lactic acid bacteria (LAB) have been extensively studied for their ability to produce extracellular polysaccharides (EPSs), which are repeating units of mono/oligosaccharides with various chemical compositions and properties. Despite the conventional belief regarding probiotics that states bacteria must be alive to exert their beneficial effects, mounting evidence opened up new perspectives on health-promoting effects of bacterial-derived molecules/metabolites. Employing EPSs seems safer alternative in complicated disorders like cancer and immune related diseases. EPSs have potential applications in textiles, cosmetology, wastewater treatment, food, and pharmaceutical industries, but little is known about their function therefore the present review aims to discuss the health-promoting properties of EPSs produced by LAB. Antioxidant, antiviral, antitumor and immunomodulating activity, with focus on the mechanisms involved, are discussed in the study. Ever-growing number of researches indicate that EPSs beneficiary effects have a promising future ahead.


Assuntos
Antineoplásicos/uso terapêutico , Fatores Imunológicos/uso terapêutico , Lactobacillales/química , Polissacarídeos Bacterianos/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Sequência de Carboidratos , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia
10.
Food Funct ; 10(4): 2186-2197, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30942219

RESUMO

A new acidic polysaccharide (GPTP-3) with a molecular weight of 2.49 × 106 Da was extracted and purified from Gynostemma pentaphyllum tea. Monosaccharide analysis revealed that GPTP-3 mainly comprised mannose (20.4%), glucuronic acid (17.4%), glucose (33.4%), and galactose (21.4%) (parentheses indicate the molar percentages). Immunostimulating assays indicated that GPTP-3 could markedly promote the secretion of NO, TNF-α, IL-1ß, and IL-6 in murine macrophage RAW264.7. TLR4 was found to be a recognized target of GPTP-3. Moreover, TLR4-related mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/Akt, including ERK, JNK, p38, and Akt, were rapidly activated by GPTP-3 in RAW264.7 cells. Furthermore, GPTP-3 was found to induce the nuclear translocation of NF-κB subunit p65. All these findings suggest that MAPK, PI3K/Akt, and NF-κB pathways are involved in GPTP-3-induced macrophage activations, and GPTP-3 has the potential to be developed as a functional food with immunomodulatory functions.


Assuntos
Gynostemma/química , Fatores Imunológicos/farmacologia , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Animais , Fatores Imunológicos/química , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/química , Polissacarídeos/química , Células RAW 264.7 , Chás de Ervas/análise , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
11.
Eur J Med Chem ; 169: 29-41, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30852385

RESUMO

Increasing evidences suggested that cisplatin can be involved in a tumor-specific immune response as an immunomodulator to improve antitumor immunity, but the designation and development are limited. Here, we report a series of novel Pt(IV) complexes derived from the conjugation of platinum(II) anticancer agents with an immune checkpoint TDO inhibitor. These complexes not only showed significant cytotoxic effects on the tested cancer cell lines, but also could enhance antitumor immune response. Particularly, complex T2, the mono-conjuagte of oxoplatin and (E)-4-oxo-4-(3-(2-(pyridin-3-yl)vinyl)-1H-indol-1-yl)butanoic acid, displayed 35-fold more potency than cisplatin against TDO-overexpressed HepG-2 cancer cells. Further study indicated that T2 could inhibit TDO via releasing a derivative of a TDO inhibitor and block kynurenine production, resulting in T-cell activation and proliferation in vitro. In vivo tests proved that T2 as a potent immunomodulator could highly promote the antitumor activity of cisplatin and effectively suppress the expression of TDO. This immunochemotherapeutic strategy can be promisingly applied to treat with TDO-overexpressed cancers.


Assuntos
Antineoplásicos Imunológicos/farmacologia , Inibidores Enzimáticos/farmacologia , Fatores Imunológicos/farmacologia , Compostos Organoplatínicos/farmacologia , Triptofano Oxigenase/antagonistas & inibidores , Animais , Antineoplásicos Imunológicos/síntese química , Antineoplásicos Imunológicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Humanos , Fatores Imunológicos/química , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Neoplasias Hepáticas Experimentais/metabolismo , Neoplasias Hepáticas Experimentais/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Estrutura Molecular , Compostos Organoplatínicos/síntese química , Compostos Organoplatínicos/química , Relação Estrutura-Atividade , Triptofano Oxigenase/metabolismo
12.
Phytomedicine ; 57: 364-376, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30831485

RESUMO

BACKGROUND: Rice callus suspension culture (RCSC) has been shown to exhibit potent antiproliferative activity in multiple cancer cell lines. RCSC and its bioactive compounds can fill the need for drugs with no side effects. HYPOTHESIS/PURPOSE: The anti-inflammatory potential of RCSC and its bioactive fractions on normal colon epithelial cell lines, was investigated. STUDY DESIGN: Three cell lines, InEpC, NCM356 and CCD841-CoN were treated with proinflammatory cytokines followed by RCSC. Cytoplasmic and nuclear ROS were assayed with fluorescent microscopy and flow cytometer. Expression analysis of immune-related genes was performed in RCSC-treated cell lines. RCSC was fractionated using column chromatography and HPLC. Pooled fractions 10-18 was used to test for antiproliferative activity using colon adenocarcinoma cell line, SW620 and anti-inflammatory activity using CCD841-CoN. Mass spectrometric analysis was performed to identify candidate compounds in four fractions. RESULTS: RCSC treatment showed differential effects with higher cytoplasmic ROS levels in NCM356 and CCD841-CoN and lower ROS levels in InEpC. Nuclear generated ROS levels increased in all three treated cell lines. Flow cytometry analysis of propidium iodide stained cells indicated mitigation of cell death caused by inflammation in RCSC treated groups in both NCM356 and CCD841-CoN. Genes encoding transcription factors and cytokines were differentially regulated in NCM356 and CCD841-CoN cell lines treated with RCSC which provided insights into possible pathways. Analysis of pooled fractions 10-18 by HPLC identified 8 peaks. Cell viability assay with fractions 10-18 using SW620 showed that the number of viable cells were greatly reduced which was similar to 6X and 33X RCSC with very little effect on normal cells which similar to 1X RCSC. RCSC fractions increased nuclear and cytoplasmic ROS vs. both untreated and inflammatory control. Analysis of four fractions by mass spectrometry identified 4-deoxyphloridzin, 5'-methoxycurcumin, piceid and lupeol as candidate compounds which are likely to be responsible for the antiproliferative, anti-inflammatory and immune-regulating properties of RCSC. CONCLUSION: RCSC and its fractions showed anti-inflammatory activity on inflamed colon epithelial cells. Downstream target candidate genes which are likely to mediate RCSC effects were identified. Candidate compounds responsible for the antiproliferative and anti-inflammatory activity of RCSC and its fractions provide possible drug targets.


Assuntos
Anti-Inflamatórios/farmacologia , Antineoplásicos/farmacologia , Fatores Biológicos/farmacologia , Fatores Imunológicos/farmacologia , Doenças Inflamatórias Intestinais/tratamento farmacológico , Oryza/citologia , Técnicas de Cultura de Tecidos/métodos , Adenocarcinoma , Anti-Inflamatórios/imunologia , Antineoplásicos/química , Fatores Biológicos/química , Morte Celular/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Neoplasias Colorretais , Citocinas/genética , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Fatores Imunológicos/química , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/patologia , Oryza/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fatores de Transcrição/genética
13.
Carbohydr Res ; 476: 44-52, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30897368

RESUMO

When grown in a semi-defined medium, L. fermentum Lf2 synthesizes significant quantities (∼2 g/L) of two exopolysaccharides (EPS). The two EPS were separated by preparative size exclusion chromatography to give a high molecular mass ß-glucan (1.23 × 106 Da) and a medium molecular mass heteroglycan (8.8 × 104 Da). The structure of the high molecular mass ß-glucan was determined using a combination of NMR spectroscopy, monomer and linkage analysis. The EPS has the following structure: The immunomodulatory activity of the high molecular mass EPS was studied in peripheral blood mononuclear cells (PBMC). Exposure of PBMC to an aqueous solution of the EPS for 24 h led to increased cell proliferation, changes in expression of the cytokines CD14 and TLR2, and to an increase in production of TNF-α compared to controls. In contrast, when cells that had been treated with EPS for 24 h and from which the EPS had been removed, were subsequently exposed to the bacterial antigen LPS very low levels of TNF-α production were observed. This result indicates that the EPS imparts immunotolerance in PBMC. An ability to modulate the release of the proinflammatory mediators, such as TNF-α, is an important goal in the development of therapies for the treatment of diseases, such as Crohn's disease and ulcerative colitis, associated with excessive release of inflammatory mediators.


Assuntos
Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Lactobacillus fermentum/química , beta-Glucanas/química , beta-Glucanas/farmacologia , Humanos , Fatores Imunológicos/isolamento & purificação , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Receptor 2 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , beta-Glucanas/isolamento & purificação
14.
Int J Biol Macromol ; 130: 399-406, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30822473

RESUMO

The lectin from Latiporus sulphureus (LSL) was purified by chromatography. Five components were obtained, of which LSL4 was the most effective one to promote cell growth, which increased cell viability to 2.6 times and 0.8 times of negative control and positive control group, so LSL4 with the highest immunomodulatory activity was selected for further identification. In addition, its immune effects on murine macrophages in vitro were evaluated. After LSL4 treatment, cell viability and phagocytic activity were greatly improved, and NO amount could reach about 24.795 ±â€¯1.559 µmol·L-1. The contents of TNF-α and IL-10 were 4.0 and 3.2 times higher than those of the negative control group, and the contents of IL-1ß and IL-6 were 2.8 and 4.1 times higher than those of the negative control group, respectively. TAK242 significantly inhibited the secretion of TNF-α, IL-1ß, IL-6 and IL-10, with inhibition rates of 53%, 61%, 48% and 54%, respectively. The results showed that 76.0 kD glycoprotein could promote cell proliferation and phagocytosis, and activate cells to release more nitric oxide, nitric oxide synthase and cytokines. The lectin has potential immunopotentiation in functional foods and pharmacology.


Assuntos
Agaricales/química , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Lectinas/química , Lectinas/farmacologia , Sequência de Aminoácidos , Animais , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Fatores Imunológicos/isolamento & purificação , Lectinas/isolamento & purificação , Macrófagos/efeitos dos fármacos , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fagocitose/efeitos dos fármacos , Células RAW 264.7 , Análise Espectral
15.
Carbohydr Polym ; 214: 44-52, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30926006

RESUMO

Two neutral polysaccharides named POP-1 and PCP-1 were purified from Polygonatum odoratum and P. cyrtonema, respectively. Results showed that both of them were composed of fructose and glucose, and the average molecular weights of them were about 5 kDa. FT-IR, 1D- and 2D-NMR analyses revealed that both POP-1 and PCP-1 contained a (2→1)-linked ß-d-fructofuranose (Fruf) backbone and (2→6)-linked ß-d-Fruf side chains with an internal α-d-glucopyranose (Glcp) in neokestose form. The difference between these two purified polysaccharides were that PCP-1 possessed of an acetyl group attached at O-3 of α-d-Glcp residue. In addition, PCP-1 exhibited a little better immune stimulating activity than POP-1 on cell viability and IL-6 production of RAW 264.7 macrophages. These results indicated that the acetyl group might affect the immunoregulatory activity and PCP-1 have more potential to be explored as an immunomodulatory agent.


Assuntos
Frutanos/farmacologia , Fatores Imunológicos/farmacologia , Polygonatum/química , Animais , Sequência de Carboidratos , Sobrevivência Celular/efeitos dos fármacos , Frutanos/química , Frutanos/isolamento & purificação , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Interleucina-6/metabolismo , Camundongos , Peso Molecular , Fagocitose/efeitos dos fármacos , Células RAW 264.7
16.
Nat Commun ; 10(1): 1012, 2019 03 04.
Artigo em Inglês | MEDLINE | ID: mdl-30833557

RESUMO

Amphiphilicity in ɑ-helical antimicrobial peptides (AMPs) is recognized as a signature of potential membrane activity. Some AMPs are also strongly immunomodulatory: LL37-DNA complexes potently amplify Toll-like receptor 9 (TLR9) activation in immune cells and exacerbate autoimmune diseases. The rules governing this proinflammatory activity of AMPs are unknown. Here we examine the supramolecular structures formed between DNA and three prototypical AMPs using small angle X-ray scattering and molecular modeling. We correlate these structures to their ability to activate TLR9 and show that a key criterion is the AMP's ability to assemble into superhelical protofibril scaffolds. These structures enforce spatially-periodic DNA organization in nanocrystalline immunocomplexes that trigger strong recognition by TLR9, which is conventionally known to bind single DNA ligands. We demonstrate that we can "knock in" this ability for TLR9 amplification in membrane-active AMP mutants, which suggests the existence of tradeoffs between membrane permeating activity and immunomodulatory activity in AMP sequences.


Assuntos
Anti-Infecciosos/química , Anti-Infecciosos/imunologia , Peptídeos Catiônicos Antimicrobianos/química , DNA/química , Receptor Toll-Like 9/química , Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Morte Celular/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Simulação por Computador , DNA/imunologia , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/imunologia , Ligantes , Macrófagos/efeitos dos fármacos , Modelos Moleculares , Conformação Proteica em alfa-Hélice/fisiologia , Espalhamento de Radiação , Receptor Toll-Like 9/imunologia , Difração de Raios X
17.
J Agric Food Chem ; 67(13): 3796-3810, 2019 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-30864792

RESUMO

In this study, Alg-Gly-Ala-Val-Leu-His (RGAVLH) obtained from pine nut ( Pinus koraiensis Sieb. et Zucc.) protein was chosen to investigate the phenomenon of immunomodulatory activity improvement upon pulsed electric field (PEF) processing. The influence of electric field intensity on immunomodulatory activity of RGAVLH was evaluated using RAW 264.7 cells. It was found that RGAVLH can not only significantly ( p < 0.05) improve the capability of macrophage phagocytosis but also promote the production of nitric oxide. RGAVLH treated under an electric field intensity of 40 kV/cm exhibited the best immunomodulatory activity. The primary and secondary structures of PEF-treated peptides were analyzed by mid-infrared (MIR) spectroscopy, Raman spectroscopy, circular dichroism spectroscopy, and one-dimensional/two-dimensional nuclear magnetic resonance spectroscopy. After PEF treatment, the primary structure of RGAVLH was not influenced, as evaluated by MIR and Raman spectra. In addition, the content of ß-sheet was decreased and active hydrogen was changed in PEF-treated RGAVLH solution. Moreover, the long-range connectivity between CαH (3.39 ppm) and NαH (8.24 ppm) was enhanced by PEF. Therefore, the improvement of the immunomodulatory activity of RGAVLH might result from the changes of the spatial state and spatial force in the peptide solution system.


Assuntos
Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Pinus/química , Animais , Eletricidade , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Nozes/química , Fagocitose/efeitos dos fármacos , Estrutura Secundária de Proteína , Células RAW 264.7
18.
Curr Top Med Chem ; 19(3): 180-185, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30854972

RESUMO

Oncology immunotherapy has gained significant advances in recent years and benefits cancer patients with superior efficacy and superior clinical responses. Currently over ten immune checkpoint antibodies targeting CTLA-4 and PD-1/PD-L1 have received regulatory approval worldwide and over thousands are under active clinical trials. However, compared to the rapid advance of Monoclonal Antibody (mAb), studies on immunotherapeutic small molecules have far lagged behind. Small molecule immunotherapy not only can target immunosuppressive mechanisms similar to mAbs, but also can stimulate intracellular pathways downstream of checkpoint proteins in innate or adaptive immune cells that mAbs are unable to access. Therefore, small molecule immunotherapy can provide an alternative treatment modality either alone or complementary to or synergistic with extracellular checkpoint mAbs to address low clinical response and drug resistance. Fortunately, remarkable progress has achieved recently in the pursuit of small molecule immunotherapy. This review intends to provide a timely highlight on those clinically investigated small molecules targeting PD-1/PD-L1, IDO1, and STING. The most advanced IDO1 inhibitor epacadostat have been aggressively progressed into multiple clinical testings. Small molecule PD-1/PD-L1 inhibitors and STING activators are still in a premature state and their decisive application needs to wait for the ongoing clinical outcomes. Since no small molecule immunotherapy has been approved yet, the future research should continue to focus on discovery of novel small molecules with distinct chemo-types and higher potency, identification of biomarkers to precisely stratify patients, as well as validation of many other immune-therapeutic targets, such as LAG3, KIRs, TIM-3, VISTA, B7-H3, and TIGIT.


Assuntos
Fatores Imunológicos/farmacologia , Imunoterapia , Neoplasias/imunologia , Neoplasias/terapia , Bibliotecas de Moléculas Pequenas/farmacologia , Anticorpos Monoclonais/imunologia , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/uso terapêutico , Proteínas de Membrana/antagonistas & inibidores , Estrutura Molecular , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/uso terapêutico
19.
Eur J Med Chem ; 170: 87-98, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30878834

RESUMO

By means of docking studies, seventeen compounds T.1-T17 have been designed and evaluated as multitarget inhibitors of VEGFR-2 and PD-L1 proteins in order to overcome resistance phenomena offered by cancer. All these designed molecules display a urea moiety as a common structural feature and eight of them (T.1-T8) further contain a 1,2,3-triazol moiety. The antiproliferative activity of these molecules on several tumor cell lines (HT-29, MCF-7, HeLa, A549, HL-60), on the endothelial cell line HMEC-1 and on the non-tumor cell line HEK-293 has been determined. The urea derivatives were also evaluated for their antiangiogenic properties, whereby their ability to inhibit tubulogenesis and kinase activity employing flow cytometry, ELISA, immunofluorescence and western blot techniques was measured. In addition, these techniques were also employed to investigate the immunomodulator action of the synthetic compounds on the inhibition of PD-L1 and c-Myc proteins. Compound T.2, 1-(3-chlorophenyl)-3-(2-(4-(4-methoxybenzyl)-1H-1,2,3-triazol-1-yl)ethyl)urea, has shown similar results to sorafenib in both down-regulation of VEGFR-2 and inhibition of the kinase activity of this receptor. Furthermore, compound T.14, (E)-1-(4-chlorophenyl)-3-(3-(4-methoxystyryl)phenyl)urea, improves the effect of T.2 as regards tube formation of endothelial cells and inhibition of VEGFR-2 tyrosine kinase activity. In addition, T.14 improves the effect of the experimental drug BMS-8 in the inhibition of PD-L1 and c-Myc proteins.


Assuntos
Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Ureia/análogos & derivados , Ureia/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Inibidores da Angiogênese/síntese química , Antineoplásicos/síntese química , Bibenzilas/síntese química , Bibenzilas/química , Bibenzilas/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Células HEK293 , Humanos , Fatores Imunológicos/síntese química , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Simulação de Acoplamento Molecular , Neoplasias/tratamento farmacológico , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Ureia/síntese química , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
20.
Int J Biol Macromol ; 128: 724-731, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30703419

RESUMO

A new polysaccharide named GRP (Glehniae radix polysaccharide) was isolated and purified from Glehniae radix by hot water extraction, ethanol precipitation, anion-exchange and gel-filtration chromatography. GRP was homogeneous, with a molecular weight of 1.33×104Da, as determined by high-performance size-exclusion chromatography-refractive index detector analysis. Its structural characteristics were investigated and elucidated by methylation analysis, gas chromatography mass spectrometry, Fourier transform infrared and nuclear magnetic resonance spectroscopy. Based on obtained data, GRP was found to be α­d­glucan containing (1→6)-linked and (1→3)-linked backbone with a branch of one (1→6)-linked and terminal glucoses submitting at the C-4 position every fourteen residues. The biological activities of GRP upon proliferation of splenic lymphocyte, RAW264.7 cells and A549 cell, and production of nitric oxide (NO) were investigated in vitro. The results showed that GRP exhibited inhibition against A549 cells proliferation and NO production in RAW264.7 cells, and displayed promotion for proliferation of mouse spleen lymphocytes and RAW264.7 cells, which suggested that GRP may have potential immunoregulation, anti-inflammatory and anti-tumor activity.


Assuntos
Apiaceae/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Água/química , Células A549 , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Camundongos , Monossacarídeos/análise , Óxido Nítrico/biossíntese , Polissacarídeos/isolamento & purificação , Células RAW 264.7 , Solubilidade , Baço/imunologia
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