Patent analysis of new target oral drugs for type 2 diabetes mellitus / 中国药房

OBJECTIVE To analyze the patents of new target oral drugs for type 2 diabetes mellitus （T2DM）， and to provide references for the research and development direction and patent layout of new domestic diabetes drugs. METHODS Based on global patent data in the HimmPat database， from multiple perspectives such as the number of patent applications and authorization， development trend， regional distribution and main applicants， statistics and analysis were performed for the patents related to 3 types of new target oral drugs for T2DM， such as glucokinase activator （GKA）， protein tyrosine phosphatase 1B inhibitor （PTP-1B-IN）， and 11β-hydroxysteroid dehydrogenase 1 inhibitor （11β-HSD1-IN）. RESULTS & CONCLUSIONS A total of 1 649 patents of GKA， 709 patents of PTP-1B-IN， 592 patents of 11β-HSD1-IN were obtained， the main applicants were well-known pharmaceutical companies， which possessed the core patents of pharmaceutical compounds. The research on GKA drugs was more mature， with a larger number of patent applications and a more comprehensive enterprise layout. Domestic enterprises， universities and research institutions had advantages in the field of PTP-1B-IN. Domestic enterprises and research institutions can leverage the advantages of traditional Chinese medicine and resources to enhance their research capabilities and improve technological competitiveness through core technology exploration， the exploration of process route， patent layout， industry- university-research cooperation and the establishment of patent pool.

Integration of network and experimental pharmacology to decipher the antidiabetic action of Duranta repens L / 中西医结合学报

OBJECTIVE@#Duranta repens is reported to contain a wide array of secondary metabolites, including α-amylase and α-glucosidase inhibitors, and - has potent antioxidant activity. The present study evaluated the network pharmacology of D. repens (whole plant) with targets related to diabetes mellitus and assessed its outcome by evaluating the effects of the hydroalcoholic extract of D. repens in streptozotocin-nicotinamide-induced diabetes mellitus in rats.@*METHODS@#Phytoconstituents of D. repens were retrieved from an open-source database and published literature, and their targets were predicted for diabetes mellitus using BindingDB and the therapeutic target database. Protein-protein interaction was predicted using STRING, and pathways involved in diabetes mellitus were identified using the Kyoto Encyclopedia of Genes and Genomes pathway browser. Druglikeness, ADMET profile (absorption, distribution, metabolism, excretion and toxicity) and cytotoxicity of compounds modulating proteins involved in diabetes were predicted using MolSoft, admetSAR2.0 and CLC-Pred, respectively. The interaction network among phytoconstituents, proteins and pathways was constructed using Cytoscape, and the docking study was performed using AutoDock4.0. The hydroalcoholic extract of D. repens was evaluated using streptozotocin-nicotinamide-induced diabetes mellitus animal model for 28 d, followed by an oral glucose tolerance test. At the end of the study, biochemical parameters like glycogen content, hepatic enzymes, antioxidant biomarkers and lipid profiles were quantified. Further, the liver and pancreas were collected for a histopathology study.@*RESULTS@#Thirty-six different secondary metabolites from D. repens were identified to regulate thirty-one targets involved in diabetes mellitus, in which protein-tyrosine phosphatase 1B (PTP1B) was primarily targeted. Enrichment analysis of modulated proteins identified 12 different pathways in diabetic pathogenesis in which the phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway was chiefly regulated. The docking study found that durantanin I possessed the highest binding affinity (-8.9 kcal/mol) with PTP1B. Similarly, ADMET profiling showed that the majority of bioactive constituents from D. repens had higher human intestinal absorptivity and minimal cytotoxicity to normal cell lines, than tumor cell lines. Further, an in vivo animal study reflected the efficacy of the hydroalcoholic extract of D. repens to lower the elevated blood glucose level by stimulating insulin secretion, maintaining pancreatic β cell mass, regulating glycolysis/gluconeogenesis and enhancing the glucose uptake in skeletal muscles.@*CONCLUSION@#The present study reflected the probable network interaction of bioactive constituents from D. repens, their targets and modulated pathways, which identified the prime regulation of the PI3K-Akt signaling pathway and PTP1B protein. Modulation of PTP1B protein and PI3K-Akt signaling pathway could contribute to enhancing glucose uptake, insulin production and glycolysis and decreasing gluconeogenesis in diabetes, which was evaluated via the experimental study.

Enantiomeric pairs of meroterpenoids from Rhododendron fastigiatum / 中国天然药物

Five pairs of optically pure meroterpenoid enantiomers (1a/1b-5a/5b) and two known compounds (6 and 7) were isolated from Rhododendron fastigiatum. Compounds 1a/1b-5a/5b were resolved from naturally scalemic mixtures by chiral HPLC. Their structures were elucidated by spectroscopic methods, X-ray crystallographic experiments, and ECD analyses. Compounds 1a/1b, 2a/2b, 3b, 4a/4b, and 5a/5b were new meroterpenoids with different polycyclic systems. Two enantiomeric pairs (2a/2b and 3a/3b), 6, and 7 exhibited inhibitory effects on protein tyrosine phosphatase 1B (PTP1B) in vitro.

Protein tyrosine phosphatase 1B inhibitory activities of ursane-type triterpenes from Chinese raspberry, fruits of Rubus chingii / 中国天然药物

Protein tyrosine phosphatase 1B (PTP1B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii (Chinese raspberry) were used as a kind of dietary traditional Chinese medicine. The methanolic extract of R. chingii fruits exhibited significant PTP1B inhibitory activity. Further bioactivity-guided fractionation resulted in the isolation of three PTP1B inhibitory ursane-type triterpenes: ursolic acid (1), 2-oxopomolic acid (2), and 2α, 19α-dihydroxy-3-oxo-urs-12-en-28-oic acid (3). Kinetics analyses revealed that 1 was a non-competitive PTP1B inhibitor, and 2 and 3 were mixed type PTP1B inhibitors. Compounds 1-3 and structurally related triterpenes (4-8) were further analyzed the structure-activity relationship, and were evaluated the inhibitory selectivity against four homologous protein tyrosine phosphatases (TCPTP, VHR, SHP-1 and SHP-2). Molecular docking simulations were also carried out, and the result indicated that 1, 3-acetoxy-urs-12-ene-28-oic acid (5), and pomolic acid-3β-acetate (6) bound at the allosteric site including α3, α6, and α7 helix of PTP1B.

Flavonoids with PTP1B inhibition from Broussonetia papyrifera / 中国中药杂志

Eleven flavonoids were isolated from the twigs of Broussonetia papyrifera by column chromatography over silica gel,ODS,MCI gel,and Sephadex LH-20,as well as RP-HPLC.Their structures were identified by spectroscopic methods including NMR,MS,UV,and IR as broupapyrin A(1),5,7,3',4'-tetrahydroxy-3-methoxy-8-geranylflavone(2),8-prenylquercetin-3-methyl ether(3),broussonol D(4),broussoflavonol B(5),uralenol(6),broussonol E(7),8-(1,1-dimethylallyl)-5'-(3-methylbut-2-enyl)-3',4',5,7-tetrahydroxyflanvonol(8),broussoflavonol E(9),4,2',4'-trihydroxychalcone(10),and butein(11).Compound 1 is a new isoprenylated flavonol.Compounds 3,6,10,and 11 were obtained from the genus Broussonetia for the first time,and 4 and 7 were firstly discovered in B.papyrifera.Compounds 1-5 and 7-9 showed significant inhibitory effects on PTP1 B with IC50 values ranging from(0.83±0.30) to(4.66±0.83) μmol·L-1.

Advances in research of protein tyrosine phosphatase 1B and its inhibitors / 中国药科大学学报

Protein tyrosine phosphatase 1B(PTP1B),a member of protein tyrosine phosphatases(PTPs),plays a key role in the negative regulation of insulin and leptin signalings.Recent studies showed that PTP1B had an important connection with endoplasmic reticulum(ER)stress, pancreatic beta cells proliferation and insulin secretion,and is closely related to the pathological process of type 2 diabetes mellitus(T2DM)and obesity. Therefore,PIP1B targeted inhibitors have become a research hotspot in the treatment of these metabolic disea-ses.Based on the structural features of PTP1B and its relationship with T2DM and obesity,PTP1B inhibitors were classified according to the sites of binding.Their latest research advances were reviewed in this paper,providing a reference for the development of anti-T2DM and anti-obesity drugs targeting PTP1B.

Design,synthesis and antidiabetic activities of 5-arylmethylene thiazolidine-2,4-dione derivatives / 药学学报

In order to find highly active antidiabetic agents, the 3-amino group of skeletal structure of thiazolidine-2,4-diones (TZDs) was modified to generate the new molecules TM1 and TM2 in the present research. The new molecules TM3-TM6 containing rhodanine structural units were designed based upon the bioisostere and combination principles. The target molecules TM7, which is similar to the traditional TZDs structurally, were designed by connecting the phenolic hydroxyl of the above target molecules to carbazole through a linker. All of these target compounds were synthesized successfully by selecting suitable synthetic routes with optimized procedures. The assay results of peroxisome proliferator activated receptor response element (PPRE) agonist activity revealed that the PPAR agonist activity was decreased due to the change of TZD ring. The assay of α-glucosidase inhibitory activity and protein tyrosine phosphatase-1B (PTP-1B) inhibitory activity showed that most of the seven serials target molecules have weak activities in vitro. However, 3 of the compounds exhibited strong PTP-1B inhibitory activities. TM2-6 exhibited the highest inhibitory activities, which reached 96.71% with IC50 1.48 mg·L-1. In addition, the toxicity prediction disclosed that the highly active compounds were almost non-toxic. These results provide a hint for the development of new antidiabetic

Chemical constituents from Rubus caesius and their inhibitory activities on PTP 1B / 中草药

Objective: To investigate the chemical constituents from the stem and leaves of Rubus caesius and the inhibitory activities on PTP 1B. Methods: Compounds were separated and purified by silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative liquid chromatography. Their structures were identified by spectral methods. The PTP1B inhibitory activities were screened by microplate reader. Results: Five compounds were obtained from the stems of R. caesius respectively, elucidated as naringin (1), apigenin-7-O-β-D-glucopyranoside (2), isoquercitrin (3), hyperoside (4), and (-)-epicatechin 3-O-gallate (ECG) (5), and two compounds were obtained from the leaves respectively, elucidated as acteoside (6) and ellagic acid (7) respectively. Conclusion: Compounds 1-7 are isolated from this plant for the first time. Different fractions and compounds showed different PTP1B inhibitory activities and acteoside showed high PTP1B inhibitory activity with the IC50 value of (27.41 ± 0.61) μg/mL. This compound may be the main active composition of leaves ethyl acetate fraction.

Protein tyrosine phosphatase 1B and its effects on pancritic ß cell functions: Research advances / 国际药学研究杂志

Tyrosine phosphorylation, a key post-translational mechanism, is necessary for metabolic homeostasis. Protein tyrsine phosphatase 1B(PTP1B), the main negative regulator in insulin signaling pathway, is becoming a potential target for the treatment of type 2 diabetes mellitus and related metabolic diseases. In thin paper, the structural properties of PTP1B and its regulation on the mass, insulin secretion, endoplasmic reticulum ER stress and inflammation in pancreatic ß cells are reviewed. It is indicated that PTP1B inhibition is important for the protection of ß cells and the treatment of type 2 diabetes mellitus.

Investigating the effects of compound WS090152 on non-alcoholic fatty liver in mice / 药学学报

To investigate the effects and the mechanism of compound WS090152 on non-alcoholic fatty liver (NAFL), the compound was administrated in C57BL/6J mice fed a high fat diet at 50 mg·kg-1 by lavage. The lipid accumulation in liver was determined by the content of hepatic triglyceride (TG) and the histological pathological analysis. The levels of body weight gain, serum total cholesterol (TC) and TG were measured to evaluate lipid metabolism. Insulin sensitivity was determined by glucose infusion rate (GIR) value in hyperinsulinemic-euglycemic clamp test. The expression of related proteins in liver was measured by Western blot. The effect on the target protein tyrosine phosphatase 1B (PTP1B) was assessed by the activity of recombinate human PTP1B in vitro, and by the expressions of PTP1B in vivo, respectively. The content of hepatic TG (PPPPP50 value of 0.34 μmol·L-1; the expression of PTP1B was significantly downregulated, and the phosphorylation of its downstream insulin receptor (IR) and AKT was upregulated by WS090152 administration in the livers of NAFL mice. The expression of hepatic lipogenesis-related proteins-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC) was attenuated. These results suggest that compound WS090152 can ameliorate NAFL by increasing insulin sensitivity and decreasing hepatic lipogenesis probably through inhibition of PTP1B.

Terpenes from South China Sea soft coral Sarcophyton trocheliophrum / 中草药

Objective: To investigate the terpenes of the South China Sea soft coral Sarcophyton trocheliophrum. Methods: Subjected to 1H-NMR-guided fractionation, the chemical constituents were isolated and purified using column chromatographies on silica gel, Sephadex LH-20, and high performance liquid chromatography (HPLC). Their structures were determined through the analysis of spectroscopic data. Results: Eight diterpenes and one sesquiterpene were isolated from the ethyl ether soluble part of acetone extract of S. trocheliophrum and their structures were identified as cembrene A (1), (E,E,E)-7,8-epoxy-1-isopropyl-4,8,12- trimethylcyclotetradeca-l,3,11-triene (2), sarcophytonolide A (3), deacetylemblide (4), 4Z,12Z,14E-sarcophytolide (5), sarcrassin D (6), emblide (7), (4Z,8S,9R,12E,14E)-9-hydroxy-1-isopropyl-8,12-dimethyloxabicyclo [9.3.2]-hexadeca-4,12,14-trien-18-one (8), and β-elemene (9), respectively. Conclusion: Compound 4 is found to be a new natural product, and its 13C-NMR data are recorded for the first time. Moreover, this is the first report that compounds 1, 3, 5, and 8 are obtained from the title soft coral. In bioassay, compound 5 exhibits the moderate human protein tyrosine phosphatase 1B (PTP1B) inhibitory activity (IC50 = 15.4 μmol/L).

Isolation and Identification of Lignans Chemical Constituents from Coptis chinensis and Their Inhibitory Activity to Protein Tyrosine Phosphatase-1B / 中国药房

OBJECTIVE:To isolate and identify lignans chemical constituents from Coptis chinensis,and to investigate their in-hibitory activities to protein tyrosine phosphatase-1B (PTP1B). METHODS:The acetic ether part of 95% ethanol extract from C. chinensis was isolated by HPLC and gel column chromatography,and the structure of chemical constituents were identified by UV, IR,MS and NMR. Using RK-682 as positive drug,the inhibitory activity to PTP1B was investigated(the concentrations of posi-tive drug and isolated chemical constituents were 0,15,30,50,65,80,100,120,140,170 and 200 μmol/L,respectively). In-hibitory rate and IC50 of them were calculated. RESULTS:5 lignans were isolated,including 9-acetyl lanicepside B,Lanicepside A,Woorenogenin,(+)-isolariciresinol and(+)-lariciresinol gluciside(compound 1-5),and compound 1 and 2 were isolated from C. chinensis for the first time. Compound 1-5 inhibited PTP1B to certain extent in concentration-dependent manner. IC50 of them were 57,49,71,58 and 51 μmol/L,but their effects were not as good as RK-682 (IC50=32 μmol/L). CONCLUSIONS:5 lig-nans have been isolated from C. chinensis and can inhibit PTP1B to certain extent.

Research progress of relationship between protein tyrosine phosphatase 1B and hematologic neoplasms / 白血病·淋巴瘤

Tyrosine phosphorylation is a key post-translational mechanism that regulates cellular processes and maintains homeostasis.Aberrant changes in tyrosine phosphorylation are often associated with disease states such as metabolicdisorders,cancer and cardiovascular disease.Protein tyrosine phosphatases (PTPs) are the enzyme family that regulates protein phosphorylation level of tyrosine residues in the cellular processes and signaling ways.So far,scientists have discovered 112 kinds of human PTPs.Among them,PTP1B is widely and clearly studied.Recently,as an enzyme that play a role in oncogenesis,PTP1B has been wildey studied by scientists.Here,we highlight the relationship between protein tyrosine phosphatase 1B and hematologic neoplasms.

Research progress on natural products on new targets of type 2 diabetes / 中草药

Type 2 diabete, caused by inadequate secretion of insulin and insulin resistance in tissues and organs, is also called non insulin dependent diabete. At present, over 90% diabetes is type 2 diabete which is seriously harmful to human health, however, the current clinical application of chemical synthetic medicines have different degrees of side effects. Therefore, the study of components with antidiabetic activity acting on different targets from the natural products with low toxicity has become a focus of current research. The new targets, such as PTP1B, DPP-IV, PPAR, and AMPK, were discovered in recent years. In this paper, active components in natural products on new targets for diabetes are reviewed, so as to provide the reference for the further research of diabetes.

Analysis of protein tyrosine phosphatase 1B gene mutation in myeloproliferative neoplasms / 白血病·淋巴瘤

Objective To analyse protein tyrosine phosphatase 1B (PTP1B) gene mutation in myeloproliferative neoplasms (MPN).Methods DNA sequencing technology was used to detect DNA sequences of PTP1B in MPN patients (n =84) and normal controls (n =37).Results For Exon1-6,Exon9 and Exon10,84 cases of MPN patients and 37 cases of control group were not detected mutation.For EXON 8,18 of 84 MPN patients had Exon8 C/T heterozygous mutation and 10 of 37 normal controls were detected C/T heterozygous mutation.There was no significant difference between MPN patients and normal controls (x2 =0.453,P =0.501).Exon7 was detected in 38 MPN patients and 2 cases of patients were found C/T heterozygous mutation,while in the control group,1 case with G/C heterozygous mutation.All of the cases were not detected homozygous mutation.Conclusion Using DNA sequencing technology to detect gene mutations of PTP1B,there is no significant difference between MPN patients and normal controls.

Effect ofPTP1B gene on the proliferation and migration of human gastric cancer cell lines / 中国癌症杂志

Background and purpose:Gastric cancer is the most common malignant tumor of digestive tract, and the possibility of postoperative recurrence and metastasis is higher. Our previous studies showed that protein tyrosine phosphatase1B (PTP1B) gene is closely correlated with tumor size, lymph node metastasis and TNM stage of gastric cancer. The purpose of the present study was to explore the effect ofPTP1B gene on cell proliferation and migration of gastric cancer cell lines.Methods:Short hairpin RNA (shRNA) sequence targetingPTP1B gene and PTP1B cDNA were transfected into MKN28 and MKN45 cells, respectively. The expression of PTP1B mRNA and its protein in MKN28 and MKN45 cells were detected by quantitative real-time PCR (qRT-PCR) and Western blot, respectively. The effect of PTP1B on cell proliferation and migration was respectively assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay.Results:Compared with blank and negative controls, the expressions of PTP1B mRNA and protein in MKN28 cells were successfully suppressed after the cells were transfected with shRNA (P<0.05). As CCK-8 test showed, the proliferation of MKN28 cells was successfully restrained at 48, 72 and 96 h after transfected with shRNA compared with blank control and negative control (P<0.05). Transwell and wound healing test were performed after PTP1B expression was interfered by shRNA. The result demonstrated that migration of MKN28 cells was signiifcantly inhibited (P<0.05). On the contrary, the expressions of PTP1B mRNA and protein in MKN45 cells were obviously enhanced after the cells were transfected with PTP1B cDNA. And the proliferation and migration of cells were significantly increased.Conclusion:PTP1B gene is an important enchancer for the proliferation and migration of gastric cancer.

Effects of 3'-hydroxy puerarin on improving insulin resistance in 3T3-L1 adipocytes and their mechanisms / 中草药

Objective: To investigate the effects of 3'-hydroxy puerarin on improving insulin resistance in 3T3-L1 adipocytes and their mechanisms. Methods: The proliferation of 3T3-L1 preadipocytes was tested by MTT assay and the differentiation by oil red O staining. The insulin resistance model was induced by dexamethasone. Cellular glucose consumption was determined by GOD-POD assay and the concentration of FFA by colorimetric methods. The expression of PPARγ and PTP1B genes in insulin resistant adipocytes was analyzed by qPCR. The PPARγ-transactivation activity of 3'-hydroxy puerarin was examined by using a hybrid reporter gene assay and the activity of PTP1B by colorimetric methods. Results: Compared with the medium control group, 3'-hydroxy puerarin significantly activated PPARγ at 0.1 and 10 μmol/L (P 0.05); increased the proliferation and differentiation of 3T3-L1 preadipocytes at 1-10 μmol/L (P 0.05). Conclusion: 3'-Hydroxy puerarin can improve the insulin resistance via up-regulating the expression of PPARγ.

Protein Tyrosine Phosphatase 1B Inhibitors from Plantago asiatica / 中草药·英文版

Objective To identify the active compounds for protein tyrosine phosphatase 1B (PTP1B) from the seeds of Plantago asiatica. Methods Bioassay-guided fractionation resulted in the isolation of iridoid glucosides (1-5) with PTP1B inhibitory activity. Results Five compounds were identified as desacetylhookerioside (1), melittoside (2), geniposidic acid (3), 10-O-acetyl-geniposidic acid (4), and alpinoside (5). Conclusion Isolated compounds 3-5 inhibit PTP1B with IC50 values ranged from (16.3 ± 1.1) to (19.8 ± 1.2) μmol/L.

The role of protein tyrosine phosphatase 1B in tumorigenesis and progression / 肿瘤

Protein tyrosine phosphatase 1B (PTP1B) is a major non-transmembrane protein tyrosinephosphatase and plays an important role in signaling pathway. PTP1B also acts as an essential regulatorin numerous physiological processes and it has a vital role in cell growth, differentiation, metabolism,migration, gene transcription and apoptosis through modulating intracellular tyrosine phosphorylation.Evidence has demonstrated that PTP1B is associated with tumor. Although many conflicting resultssuggested that PTP1B has two contradictory effects (supressing or promoting ) on tumor, the real effectdepends on the substrate involved and the cellular context. This review describes different mechanismsof PTP1B in tumorigenesis and progression in breast cancer, colon cancer, hepatic carcinoma, lymphoma,ovarian cancer, esophageal cancer, prostate cancer and gastric cancer. These results further theunderstanding of PTP1B function and highlight the great prospective of PTP1B inhibitors in tumor therapy. Copyright© 2011 by TUMOR.

Association of PTP1B gene polymorphism with obesity in Chinese children / 中南大学学报(医学版)

Objective To investigate the distribution characteristics of protein tyrosine phosphatase 1B (PTP1B) gene IVS6+G82A and Pro303Pro polymorphisms in Chinese children and determine the effect of PTP1B gene IVS6+G82A and Pro303Pro polymorphisms on the pathogenesis of childhood obesity. Methods A total of 147 Chinese obese and 118 healthy children were randomly selected and enrolled to identify IVS6+G82A and Pro303Pro genotypes by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay. Waist circumference (WC), waist to hip ratio (WHR), percentage of body fat (%BF),systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting plasma glucose (FPG), serum triglycerides (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), plasma fasting insulin (FINS), homeostasis model assessment for insulin resistance (HOMA-IR), and plasma leptin were examined. Results The allele frequencies of IVS6+G82A and Pro303Pro were 59.5% and 19.4% in obese children, and 53.4% and 11.0% in healthy children, respectively. There were significant differences in allele frequencies of Pro303Pro polymorphism between the obese and the control group. Pro303Pro polymorphism was associated with body mass index, WC, TG, and LDL C in the obese subjects. There was not di fference in the genotype distributions or allele frequencies of IVS6+G82A polymorphism between the obese and the control group. Further analysis showed no association between the genotypes of IVS6+G82A and clinical characteristics in the obese subjects. The linkage disequilibrium analysis for IVS6+G82A and Pro303Pro (D′: 0.441, r2: 0.027) was weak.Conclusion PTP1B gene Pro303Pro polymorphism might be associated with the pathogenesis of obesity in children and could affect the lipid metabolism in Chinese obese children.