ABSTRACT
There are two serious obstacles to tumor immunotherapy. Firstly, the immune response of the tumor is seriously reduced due to immunosuppressive tumor microenvironment (ITM) and low immunogenicity of tumor. The second obstacle is the dense and complex heterogeneous structures, which seriously prevent the nanoparticles (NPs) from penetrating deeper into tumor tissue. Immunogenic cell death (ICD) induced by doxorubicin (DOX) is an effective method to enhance tumor immune activity. However, interferon-γ (IFN-γ) secreted by cytotoxic T lymphocytes (CTL) after ICD induction would increase the expression of indoleamine 2,3-dioxygenase 1 (IDO1) and enhance ITM. IDO1 siRNA would reduce the expression of IDO1 protein, regulate the tumor immunosuppressive microenvironment and regulate ITM, so as to enhance the ICD effect of DOX. In this paper, a novel charge conversional, particle size reduction and highly penetrable NPs based on a pH sensitive copolymer poly(ethylene glycol)-poly-L-lysine-2,3-dimethylmaleic anhydride (mPEG-PLL-DMA, PLD) and polyamidoamine (PAMAM) dendrimers to achieve deep delivery of tumor tissue. DOX and IDO1 siRNA were encapsulated to achieve efficient tumor immunotherapy. Preparation and cell level experiments showed that PLD material had significant pH sensitivity. Results of 3D tumor penetrable experiment in vitro showed that adding the pH sensitive material PLD significantly improved the permeability of the preparation. In addition, 4T1 tumor model was established for BALB/c mice and all animal experiments were displayed in according with the requirements of the Animal Experiment Ethics Committee of Shenyang Pharmaceutical University. The results of in vivo efficacy experiments and tissue experiments evaluated that IDO1 siRNA significantly improved the ICD effect owing to DOX, so as to significantly inhibit tumor growth.
ABSTRACT
Flavonoids is one of the biggest families of the plant-derived secondary metabolites with structural diversity. Until now, over 10 000 kinds of flavonoids with distinct structures have been purified and identified from plants, and some of them possess a range of important pharmacological effects, such as anticancer, anti-inflammatory and so on. So far, a number of genes and enzymes responsible for the biosynthesis of flavonoids have been reported, especially, a great of progress has been achieved in the synthetic biology of flavonoids in the recent years. Herein, based upon a brief introduction on the biosynthesis of flavonoids, this review summarizes the research advances in synthetic biology of flavonoids in the past two decades (2001-2021), highlighting the cell factories construction of the representative flavonoids. And, a brief discussion and prospects of the relevant metabolic bottlenecks and optimizing strategies are proposed.
ABSTRACT
In recent years, the use of the body's immune system for anti-tumor immunotherapy has received extensive attention. However, the immunosuppressive tumor microenvironment (TME) limits the effect of immunotherapy. Therefore, overcoming the limitations of TME and immunosuppressive cells plays an important role in tumor immunotherapy. Nano agents have great potential to reprogram the immunosuppressive microenvironment and provide an effective strategy for immunotherapy. With the continuous development of active targeting nano carrier technology and the deepening of the research on drug action sites, subcellular organ targeting nano carrier materials with more accurate active targeting function have also attracted more and more attention. This review will briefly introduce the relationship between subcellular organelles and tumor, summarize the design strategy and research progress of targeted nano drug delivery system based on the characteristics of acidity, reactive oxygen species (ROS) activity, immunogenicity, and TME of immunosuppressive cells, to provide reference for the construction of subcellular pathway targeted drug delivery system in tumor immunotherapy.
ABSTRACT
In the research and development of new drugs, it is very important to investigate the in vitro metabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant human drug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major human UDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from human liver cDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insect cell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insect cells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, 1H NMR, and 13C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insect cell expression systems constructed in this study can be used for in vitro metabolism evaluation in the early stage of new drugs research and development, and also provide a new tool for the synthesis of glucuronide metabolites.
ABSTRACT
Praeruptorin C (PC), D (PD) and E (PE) are important compounds extracted from Peucedanum praeruptorum DUNN and have been reported to exert multiple pharmacological activities. The present study is purposed to determine the inhibition of PC, PD and PE on the activity of important phase I metabolic enzymes-carboxylesterases (CES). In vitro human liver microsomes (HLM) incubation system was used to determine the inhibition potential of PC, PD and PE on the activity of CES1 and CES2. Inhibition behaviour was determined, and in vitro-in vivo extrapolation was performed by using the combination of in vitro inhibition kinetic parameter (Ki) and in vivo exposure level of PD. PD exhibited the strongest inhibition on the activity of CES1, with 81.7% activity inhibited by 100 μmol·L-1 of PD. PD noncompetitively inhibited the activity of CES1 with the Ki to be 122.2 μmol·L-1, indicating inhibition potential of PD towards CES1 in vivo. Therefore, closely monitoring the endogenous metabolic disorders caused by PD and interaction between PD and drugs mainly undergoing CES1-catalyzed metabolism is very necessary.
ABSTRACT
The glycosides of 4'-demethylepipodophyllotoxin (DMEP) possess various pharmacological activities; however, the chemical synthesis of these glycosides faces challenges in regioselectivity, stereoselectivity, and the protection and de-protection of functional groups. In this work, a novel glycosyltransferase (GT) gene AbGT5 from Aloe barbadensis was successfully cloned, heterogeneously expressed and purified. Recombinant AbGT5 was able to catalyze the glycosylation of DMEP and the glycosylated product, which was separated from the preparative scale reaction, was characterized as DMEP 4'-O-β-D-glucoside via MS, 1H-NMR, 13C-NMR, HSQC and HMBC. According to the investigations of enzyme properties, AbGT5 show the highest activity around 20 ℃ in the buffer of pH 9.0, and it was independent of divalent metal ions. Under the optimum conditions, the conversion rate of DMEP can reach 80%. Above all, in this work the enzymatic glycosylation of DMEP was achieved with high efficiency by the novel GT AbGT5.
ABSTRACT
Liposomes as a drug carrier is easy to form aggregation and cause drug leakage in vitro. In addition, the degradation and elimination in vivo happens frequently to reduce its delivery activity. Development and application of liposomes are restricted by the instability. The appropriate techniques and methods are great important in the study of pharmaceutical stability of liposomes. In this paper, the techniques and methods are reviewed on pharmaceutical stability evaluation of liposomes, which was done from physical, chemical and biological stability for the difference in stability of liposomes. The research strategies for establishing the stability evaluation system and improving the value of liposomes have been discussed to make full therapeutic advantage of it.
ABSTRACT
Each of the syndesmotic ligaments has different biomechanical effects. Minimal movement occurs in the syndesmosis during the motion of the ankle joint, and the syndesmotic injury is associated with the ankle position and the patterns of external forces. Controversy still exists with respect to the optimal fixation of the syndesmosis. Malreduction of the syndesmosis would lead to abnormal pressure distribution on the ankle joint. Therefore, the flexible fixation might more accord with the physical movement of the syndesmosis. In this article, the biomechanical studies on syndesmosis are reviewed to provide the reference for proper treatment method for syndesmotic injury.
ABSTRACT
The present study was designed to establish and optimize a new method for extracting chlorogenic acid and cynaroside from Lonicera japonica Thunb. through orthogonal experimental designl. A new ultrahigh pressure extraction (UPE) technology was applied to extract chlorogenic acid and cynaroside from L. japonica. The influential factors, including solvent type, ethanol concentration, extraction pressure, time, and temperature, and the solid/liquid ratio, have been studied to optimize the extraction process. The optimal conditions for the UPE were developed by quantitative analysis of the extraction products by HPLC-DAD in comparison with standard samples. In addition, the microstructures of the medicinal materials before and after extraction were studied by scanning electron microscopy (SEM). Furthermore, the extraction efficiency of different extraction methods and the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the extracts were investigated. The optimal conditions for extracting chlorogenic acid and cynaroside were as follows: ethanol concentration, 60%; extraction pressure, 400 MPa; extraction time, 2 min; extraction temperature, 30 °C; and the solid/liquid ratio, 1 : 50. Under these conditions, the yields of chlorogenic acid and cynaroside were raised to 4.863% and 0.080%, respectively. Compared with other extraction methods, such as heat reflux extraction (HRE), ultrasonic extraction (UE), and Sohxlet extraction (SE), the UPE method showed several advantages, including higher extraction yield, shorter extraction time, lower energy consumption, and higher purity of the extracts. This study could help better utilize L. japonica flower buds as a readily accessible source of natural antioxidants in food and pharmaceutical industries.
Subject(s)
Analytic Sample Preparation Methods , Methods , Antioxidants , Chlorogenic Acid , Chromatography, High Pressure Liquid , Flowers , Chemistry , Glucosides , Lonicera , Chemistry , Luteolin , Plant Extracts , PressureABSTRACT
Poly(β-amino esters) (PBAE) are used for drug carrier and have many advantages, such as pH-sensitivity, low toxicity, structural diversity and the synthetic method of PBAE is easy. Therefore they are possessed broad application prospect in tumor-targeted drugs delivery systems. In this paper, the structural features and target drugs delivery property of PBAE are reviewed. The application forms of PBAE and different anti-cancer drugs loaded in the copolymer for tumor-targeted drugs delivery systems are introduced particularly.
Subject(s)
Antineoplastic Agents , Chemistry , Drug Carriers , Chemistry , Drug Delivery Systems , Esters , Chemistry , Hydrogen-Ion Concentration , Neoplasms , Drug Therapy , PolymersABSTRACT
The discovery, sorting and identification methods as well as targeted drug delivery systems for cancer stem cells (CSCs) have been reviewed by consulting the recent research papers. CSCs have been believed to be responsible for the occurrence and development of chemo-resistance, leading to the failure of chemotherapy. Much progress has been made in the approaches for CSCs targeting drug delivery systems. The understanding and targeted drug delivery systems for CSCs are promising to provide an alternative for cancer therapy.
Subject(s)
Animals , Humans , Antineoplastic Agents , Pharmacology , Therapeutic Uses , Apoptosis , Drug Delivery Systems , Methods , Drug Resistance, Neoplasm , Flow Cytometry , Neoplasms , Drug Therapy , Pathology , Neoplastic Stem Cells , Pathology , Signal Transduction , Wnt Signaling PathwayABSTRACT
Hyaluronic acid (HA) as anticancer drug carrier has become the new hot point in the field of tumor-targeted drugs delivery system in recent years. Tumor therapeutic agents could be transmitted into cells because of hyaluronic acid innate ability to recognize specific cellular receptors that overexpressed on tumor cells surface. This review introduces the basic properties and physiology foundation of hyaluronic acid. Recent research developments based on different forms of HA tumor-targeted drugs delivery systems are reviewed in particular.
Subject(s)
Animals , Humans , Antineoplastic Agents , Chemistry , Therapeutic Uses , Drug Carriers , Drug Delivery Systems , Hyaluronic Acid , Chemistry , Molecular Structure , Nanoparticles , Neoplasms , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To discuss the efficacy of Leucogen tablets treatment lessen the hematological reaction and raise the efficacy therapy of interferon in chronic hepatitis B treated with PEG-alpha interferon and alpha interferon.</p><p><b>METHODS</b>A total of 395 patients with HBeAg-positive chronic hepatitis B (CHB) inpatients from January 2002 to February 2011. Group: All the patients were assigned to A or B according as during the treatment added Leucogen tablets or not.</p><p><b>RESULTS</b>(1) All of 35.9% patients had neutrophil counts decrease under 1 x 10(9)/L, A group had 29.6%, B had 42.8% patients, P = 0.01; neutrophil counts < or = 0.75 x 10(9)/L A group had 12.6% ,B group had 26.4%, P = 0.02; neutrophil counts < or = 0. 5 x 10(9)/L A group had 4.8%, B group had 16.4%, P = 0.04. (2) A group had 8.2% patients interferon-alpha dose decreased, all the patient finished the period of therapy. B group had 23.3% patients interferon-alpha dose decreased, 2.1% of patients had paused. A group had 40.3% of patients interferon-alpha beyond conventional dose, B group had only 5.2%. (3) All of 9.8% patients had hematoblast decrease under 100 x 10(9)/L, A group had 8.7%, B had 11.1% patients; hematoblast < or = 80 x 10(9)/L A group had 5.3%, B group had 7.9%; hematoblast < or = 50 x 10(9)/L A group had 1.0%, B group had 2.6%. A group had the trend of reducing hematoblast decrease. (4) At the end of therapy A group had 67.4% patients HBVDNA < 100IU/ml, 54.3% e antigen negative, 40.7% e antigen conversed; B group had 53.9%, 41.2%, 26.9%, P was respectively 0.02, 0.01, 0.01.</p><p><b>CONCLUSION</b>Leucogen tablets treatment and prevention interferon-alpha-related neutrophil counts hematological reaction in CHB treated with alpha-interferon, and had the trend of reducing interferon-alpha-related hematoblast decrease, farther improved the efficacy of alpha-interferon treatment CHB.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Antiviral Agents , Hepatitis B, Chronic , Blood , Drug Therapy , Interferon-alpha , Neutropenia , Polyethylene Glycols , Recombinant Proteins , Tablets , Thiazolidines , Therapeutic UsesABSTRACT
The dialysis method was employed to prepare blank and doxorubicin (DOX) loaded micelles formed by temperature- and pH- sensitive polyhistidine-co-polyDL-lactide-co-glycolide-co-polyethyleneglycol-co-polyDL-lactide-co-glycolide-co-polyhistidine (PHis-b-PLGA-b-PEG-b-PLGA-b-PHis). The critical micelle concentrations (CMC) of the copolymers were measured with Pyrene Fluorescent Probe Technique. The temperature- and pH- sensitive properties of the blank micelles solution were investigated by optical transmittance measurement. The morphology and diameter of DOX micelles were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The entrapment rate and drug-loading rate were determined with dialysis method. The in vitro release study was further performed to examine the temperature- and pH-responsive drug release behavior from DOX-loaded micelles. The results indicated that the CMC, entrapment efficiency and drug-loaded amount of the micelles were 7.5 x 10(-3) g x L(-1), 85.2 +/- 3.1% and 10.4 +/- 4.5%, respectively. The DOX micelle was globular-shaped with a mean diameter of 91.1 +/- 15.8 nm. The transmittance of micelle solution consistently increased with the increasing temperature or decreasing pH. In comparison to the drug release profile at physiological conditions (37 degrees C, pH 7.4), the DOX-loaded micelles showed faster drug release rate at higher temperature (41 degrees C), lower pH (pH 7.0, pH 6.5, pH 5.0) or higher temperature and lower pH (41 degrees C, pH 5.0). This indicated that the micelles showed a temperature and pH-triggered drug release pattern. Base on the above results, it can be concluded that PHis-b-PLGA-b-PEG-b-PLGA-b-PHis block copolymer micelles which respond to temperature and pH stimuli are promising smart carriers for anti-tumor drugs with the advantages of temperature- and pH- triggered drug release.
Subject(s)
Antibiotics, Antineoplastic , Chemistry , Doxorubicin , Chemistry , Drug Carriers , Drug Compounding , Histidine , Chemistry , Hydrogen-Ion Concentration , Micelles , Particle Size , Polyethylene Glycols , Chemistry , Polyglactin 910 , Chemistry , Polymers , Chemistry , TemperatureABSTRACT
Rheological properties of poloxamer 407 (brand named Pluronic F127) were examined by changing shear rate, temperature and the recovery properties of apparent viscosity after heating for several times. The results indicated that poloxamer 407 aqueous solution showed a Newtonian behavior at a low concentration while it might be a pseudoplastic fluid when the concentration reached a certain point. The thixotropy and the sol-gel transition temperature decreased with increasing the concentration (it could be an in situ gel at body temperature when the concentration of poloxamer 407 up to 15.25%). The results that obtained from the theological data would be useful in the application of poloxamer 407 such as in situ gel preparation.
Subject(s)
Dose-Response Relationship, Drug , Drug Compounding , Drug Delivery Systems , Excipients , Chemistry , Poloxamer , Chemistry , Rheology , Shear Strength , Solutions , Temperature , Viscosity , WaterABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of preoperative use of viable Bifidobacterium supplement on the intestinal flora, immune status, inflammatory response and prognosis of patients undergoing colorectal cancer resection.</p><p><b>METHODS</b>Sixty patients with colorectal cancer were randomized into treatment group (n=30) and control group (n=30). Patients in the treatment group received oral viable Bifidobacterium with routine enteral nutrition and patients in the control group received routine enteral nutrition alone. The intestinal flora of stool was analyzed and stool SIgA, serum IgG, IgM, IgA, IL-6, and C-reactive protein (CRP) were detected.</p><p><b>RESULTS</b>Postoperative Bifidobacterium/E.coli (B/E) ratio in the treatment group decreased significantly as compared to the preoperative ratio (2.01+/-0.36 vs 26.53+/-4.22, P<0.05). However, the ratios remained above one. Both preoperative and postoperative B/E ratios in the control group (0.72+/-0.14, 0.02+/-0.06) were significantly lower than those in the treatment group (P<0.05). Indexes of immunity and inflammation were not significantly different between the two groups (P>0.05). At day 9 after operation, stool SIgA was higher in the treatment group, while serum IgG, IgM, IgA, IL-6, CRP in the treatment group were lower (P<0.05). Postoperative septic complications in the treatment group was less than that in the control group (P<0.05), but other complications and hospital stay were comparable.</p><p><b>CONCLUSIONS</b>In patients with colorectal cancer, supplementation of viable Bifidobacterium before surgery can improve bacterial dysbiosis and immunity, and can reconstruct the balance of intestinal flora, and reduce infection complication of surgery.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antibodies , Blood , Bifidobacterium , C-Reactive Protein , Colorectal Neoplasms , Allergy and Immunology , Microbiology , Therapeutics , Inflammation , Interleukin-6 , Blood , Postoperative Period , Probiotics , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the efficacy and safety of ademetionine for treatment of cholestatic or mixed-type drug-induced liver disease (DILD) in children.</p><p><b>METHODS</b>The children with DILD were divided into the treated group and control group. Yinzhihuang Granule was orally administered and Compound Glycyrrhizin Injection intravenously given in patients of both groups. Those patients in the treated group were additionally treated with intravenous infusion of 250-1000 mg ademetionine for 28 d. The incidence of pruritus and adverse effects as well as biochemical parameters in all the patients and compared between the 2 groups. For statistical analysis, Chi2 test was used for between-group comparison and t test for processing the data.</p><p><b>RESULTS</b>1) Before treatment, severe pruritus was found in 17 and 16 children in the treated and control group, respectively. Two weeks after the treatment, the symptom was significantly relieved in 14 and 3 patients in the treated and control group, respectively (Chi2 = 4.52, P < 0.05). 2) As for comparisons between the 2 groups, a P value of 0.0014 for AST level was found 4 weeks, 0.045 and 0.007 for disappearance and recovery rate of jaundice, 0.0014 and 0.0006 for decrease in TBA level and 0.0003 for gammaGT level 2 and 4 weeks after the treatment.</p><p><b>CONCLUSION</b>Intravenous administration of ademetionine is safe in children with DILD and it can effectively alleviate pruritus, promote the recovery of various biochemical parameters and fasten liver functional recovery in these children. Therefore, ademetionine can be widely used for treatment of intrahepatic cholestasis in children.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Chemical and Drug Induced Liver Injury , Drug Therapy , S-Adenosylmethionine , Therapeutic Uses , Treatment OutcomeABSTRACT
It is generally believed that liposomes modified with polyethylene glycol (PEG) have no or lower immunogenicity. However, based on many recent literatures, when the PEGylated liposomes were repeatedly applied to the same animal, the immune responses occurred. The first injection of PEGylated liposomes resulted in a reduction in the circulation time and an increase in hepatic and splenic accumulation of the second dose of PEGylated liposomes in a time-interval, which was called "accelerated blood clearance (ABC)" phenomenon. Such immunogenicity of PEGylated liposomes presents a barrier in the research of liposomal formulations and their use in the clinics. This review focused on the definition, the method of verification, the development of the reason for ABC phenomenon, influencing factors of ABC phenomenon, and discussed if other PEGylated nanocarriers also induce ABC phenomenon.
Subject(s)
Animals , Antibiotics, Antineoplastic , Pharmacokinetics , Doxorubicin , Pharmacokinetics , Drug Carriers , Immunoglobulin M , Blood , Liposomes , Blood , Pharmacokinetics , Liver , Metabolism , Metabolic Clearance Rate , Particle Size , Polyethylene Glycols , Metabolism , Pharmacokinetics , Serum Albumin, Bovine , Pharmacokinetics , Spleen , Allergy and Immunology , MetabolismABSTRACT
The dialysis method was employed to load adriamycin into the micelles formed by temperature and pH sensitive polyhistidine-co-DL-lactide-co-glycolide-polyethylene glycol poly DL-lactide-co-glycolide-co-histidine (OLH-b-PLGA-b-PEG-b-PLGA-b-OLH). The critical micelle concentration (CMC) of the copolymer was measured with pyrene fluorescent probe method under different temperatures. The entrapment rate and drug-loading rate were determined with dialysis method. The diameter, morphology and surface potential of the copolymer micelles were investigated by corresponding instruments, respectively. The release behavior of adriamycin from copolymer micelles and the pH sensitivity were studied. The CMC of the copolymers ranged from 0.022 4 to 0.001 7 microg x mL(-1). The entrapment rate and drug-loading rate were 92.8% and 15.7%, respectively. The micelles have a mean diameter of (61.7 +/- 13.4) nm, and zeta potential was -9.88 mV. The in vitro adriamycin release rate increased with the pH dropping from 7.4 to 5.0. The results indicated that the CMC of the copolymers decreased as the raising of temperature, drug release behavior from the micelles possessed clearly pH sensitivity, and the copolymers may have a potential in targeted delivery system for anticancer drugs.
Subject(s)
Doxorubicin , Chemistry , Drug Carriers , Hydrogen-Ion Concentration , Micelles , Polyethylene Glycols , Chemistry , Polyglactin 910 , Chemistry , Technology, Pharmaceutical , Methods , TemperatureABSTRACT
Artificial neural network (ANN) is a multi-objective optimization method that needs mathematic and statistic knowledge which restricts its application in the pharmaceutical research area. An artificial neural network parameters optimization software (ANNPOS) programmed by the Visual Basic language was developed to overcome this shortcoming. In the design of a sustained release formulation, the suitable parameters of ANN were estimated by the ANNPOS. And then the Matlab 5.0 Neural Network Toolbox was used to determine the optimal formulation. It showed that the ANNPOS reduced the complexity and difficulty in the ANN's application.