Characterization and Applications of Single-Domain Antibody Mimics against ErbB3

As a member of EGFR family, ErbB3 is a critical heterodimerization partner of EGFR and ErbB2 in many EGFR- or ErbB2-driven cancers. As a ligand of ErbB3, heregulin β-1 (NRG1) induces the dimerization of ErbB3 with other ErbB family members and triggers downstream pathways. Using the directed protein evolution technique mRNA display, we successfully identified SDAHER3-A1 and SDAHER3-D5 that bind to ErbB3 with very high affinity and specificity in vitro and on cell surface. A biparatopic SDAHER3-D5A1 was constructed for improved biophysical properties and picomolar affinity (80.3 ± 20.3 pM) to ErbB3. SDAHER3-D5A1 binds to ErbB-expressing MCF7 cells with a binding affinity of 35.61 ± 17.07 nM. SDAHER3-D5A1 inhibits the NRG1-induced proliferation in a dose-dependent manner. SDAHER3-D5A1 is able to carry the conjugated AuNP to ErbB3-overexpressed cell and kill the cancer cell by hyperthermia treatment. Combined with Lapatinib, a kinase inhibitor, SDAHER3-D5A1 showed improved efficiency in inhibiting the proliferation of cancer cells with ErbB2 and ErbB3 overexpressed. An examination of downstream signaling pathways suggested that SDAHER3-D5A1 significantly decreased NRG1-induced ErbB3-/Akt-/ERK- phosphorylation and the combination with the drug further inhibits the phosphorylation. SDAHER3-D5A1-IRDye800 conjugates was synthesized and confirmed that SDAHER3-D5A1 can be used to target ErbB3-overexpressed tumor xenograft models and may be very useful for imaging purpose.Master of Scienc

PO-257 Application of biochemical index monitoring in Chinese hearing-impaired table tennis team

  Objective Using the theory of sports biochemistry, we can better complete the formulation of the training plan, the evaluations of training effect and the athlete's fatigue status, the real-time monitoring of training intensity and training amount as well as the function level of athletes and the comprehensive evaluation of nutritional status. To provide some theoretical guidance for the daily training of Chinese hearing-impaired table tennis team and to gradually establish the biochemical monitoring model for the Chinese Deaf Table tennis team, this subject has applied biochemical index monitoring to this team. Methods 2.1 Research Object 8 Athletes (male 4 female 4) of the Chinese hearing-impaired table tennis team were studied. The average height, weight, and age were 174.3±5.1cm, 65.4±11.6kg, 23.5±4.9 years respectively.2.2 Research Methods 2.2.1 Literature To find out the energy-supplying and metabolic characteristics of table tennis as well as the modes to accelerate the recovery period of body fatigue, we have searched a variety of literature on this purpose. The investigation of athletes' nutrition regulation, physical training and rehabilitation and other related research results lays a good foundation for the implementation of the project. 2.2.2 Expert Interview This paper discusses the energy metabolism characteristics of the hearing-impaired table tennis players by discussing with the coaches, athletes and experts and scholars of the national Hearing Impairment project group. The evaluation index and standard of athlete's body function, the method of training load monitoring, the way of nutrition regulation, the means of fatigue recovery and the process of physical training were discussed. 3.2.3 Field Investigation From April, 20th, 2017 to July, 10th, 2017, two researchers have long been in the center of national table tennis training located at Zheng Ding, Hebei province, to complete training monitoring and research services. Results 3 Research Results Phlebotomizing 7 of the Chinese hearing-impaired table tennis players in June,2nd,2017 and June 7th, 2017 respectively, The blood samples were analyzed by the team of experts and researchers in the Sports Physiology and Biochemistry Laboratory in Beijing Sport University. Part of the results was shown below in table 1, table 2. (1) Training intensity monitoring Huang Mengping had a slightly higher creatine kinase, a slight decrease in testosterone, a slight rise in cortisol, a lower testosterone/cortisol value, as well as an insignificant reduction in body function than the previous time. She should focus on the recovery of her body condition after training. In general, the testosterone, cortisol and testosterone/cortisol level in this team is slightly higher than the last time, blood urea lower than the previous. Strengthening the monitoring of exercise and nutrition and continuing to maintain a high body synthesis ability are favorable to intense competition. The overall creatinine level decreased, whereas 4 members of the team had insignificantly high creatinine, especially for Huang Mengping who first had appeared high level of it. Although this condition was not too severe to focus on, we should strengthen the regulation of athlete muscle injury and protect their kidney functions. (2) Monitoring of endocrine system indicators Both males and females had higher testosterone synthesis capabilities. They should continue to take the nutritional supplements. The T/C ratio of Tian Jiping has escalated sharply, but it was lower than that of other players. We should pay attention to his training load and promote the recovery training. (3) Monitoring of Iron Metabolic Index Huang Mengping's hemoglobin value and RBC value have been slightly elevated this time. It is suggested that she should keep on to take nutritional supplements and strengthen aerobic endurance training. Hemoglobin for Wang Zhe and Lin Huan (slightly decreased this time) was higher than their counterparts. The ability to transport oxygen was excellent, and it is agreeable to keep on. They need to continue to maintain and promote the body's oxygen reserve capacity. At the same time, they can also upgrade their overall red blood cell levels. Men have seen a high level of hemoglobin and the number of red blood cells as well as robust capabilities of oxymoron reservation and transportation. It is recommended to enhance aerobic endurance training, mainly improving the aerobic capacity. The serum iron of Tian Jiping was promoted quickly. He needs to build up the amount of iron nutrient intake and support the body synthesis. The full iron metabolic indicators were normal, but Zhang Chaoyue and Lin Lin Huan have been seen the lack of the abilities to synthesis iron. So we need to pay attention to iron metabolism-related nutrition intake. In general, iron protein level and the capability to reserve iron have declined. It is recommended to take more iron nutrients, Especially for Shi Ce. (4) Food surveillance After the diet regulation, the indicators for the team's blood fat were within the normal range. Zhang Chaoyue and Wang Zhe should control the high-fat food intake, thus reducing the amount of low-density lipoprotein. (5) Surveillance of immune Indicators The white cell, blood cell classification and immunoglobulin (IgG, IgM, IgA) of the team are all normal, Wang Zhe’s immunoglobulin A and Xu You’s immunoglobulin M are slightly low, we should pay attention to protection. Conclusions  Through the overall monitoring and analysis of the biochemical indexes of the leading athletes of Chinese hearing-impaired table tennis team, The characteristics of biochemical indexes of the high-level hearing-impaired table tennis players are mastered. On the basis of the overall characteristics of the individual, we carried out personalized monitoring and evaluation to give coaches and athletes advice on training, rehabilitation and other aspects related to practice. Biochemical tracking in China's hearing-impaired table tennis team has achieved an excellent application results that have been unanimously recognized by the coaches and athletes. &nbsp

A study of conjugate addition of curcumin and chalcone derivatives

Curcumin is one of the promising herbal-based drugs. It has been shown to have antioxidant,antibacterial, anti-angiogenic and other activities. As curcumin's derivative, chalcone shares similar functions. Both of these two compounds have alpha,beta-unsaturated carbonyl structures(enone), which is a typical 1,4-conjugate addition (Michael addition) acceptor. Glutathione is an endogenous tripeptide, whose sulfhydral group is a typical nucleophilic agent. In this case, the derivatives of curcumin and chalcone may be reduced by glutathione and their pharmacological functions would be changed. The study focused on how to use quantum chemistry tools and transition state theory to access to the conjugate addition of alpha,beta-unsaturated carbonyl compounds. Besides, the reductions of the derivatives of chalcone were also studied. The characteristics of the reactions were obtained by analyzing geometries and energy profiles of the simplified reactions, as well as the influence of functional groups on derivatives in this type of reaction. This study may be generally useful for the scientific community for two fundamental reasons: (a) to provide general strategies to enhance or retard drugs from reacting with glutathione, and (b) to provide insight into computational methods that are able to help design potential lead candidates

Local and transient gene expression primes the liver to resist cancer metastasis

The liver is the primary site of metastasis for gastrointestinal cancers and is a location highly susceptible to the establishment of metastasis in numerous other primary cancers, including breast, lung, and pancreatic cancers. The current standard of care typically consists of primary tumor resection and systemic administration of potent but toxic chemotherapeutics, yielding a minimal improvement in the median survival rate. CXCL12, a chemokine, is a key factor for activating the migration/survival pathways of CXCR4+ cancer cells and for recruiting immunosuppressive cells to areas of inflammation. Therefore, reducing CXCL12 concentrations within the liver has the potential to decrease tumor and immunosuppressive cell activation/migration within the liver. However, because of off-target toxicities associated with systemic administration of anti-CXCL12 therapies, transient and liver-specific expression of a CXCL12 trap is necessary. To address this challenge, we developed a lipid calcium phosphate nanoparticle optimized for delivering plasmid DNA, encoding an engineered CXCL12 protein trap, to the nucleus of liver hepatocytes. This pCXCL12-trap formulation yielded transient (4 days) liver-specific expression, which greatly decreased the occurrence of liver metastasis in two aggressive liver metastasis models, including colorectal [CT-26(FL3)] and breast (4T1) cancers. Subsequent studies in an aggressive human colorectal liver metastasis model (HT-29) decreased the establishment of liver metastasis more effectively than did systemic administration of the CXCL12 protein trap and to a level comparable to a high-dose regimen of a potent CXCR4 antagonist (AMD3100)

Radiolabeled 6-(2, 3-dichlorophenyl)-N4-methylpyrimidine-2, 4-diamine (TH287): A potential radiotracer for measuring and imaging MTH1

MTH1 (MutT homolog 1) or NUDT1 (Nudix Hydrolase 1), also known as oxidized purine nucleoside triphosphatase, has potential as a biomarker for monitoring cancer progression and quantifying target engagement for relevant therapies. In this study, we validate one MTH1 inhibitor TH287 as a PET MTH1 radiotracer. TH287 was radiolabeled with tritium and the binding of

Pseudo-Cyclization through Intramolecular Hydrogen Bond Enables Discovery of Pyridine Substituted Pyrimidines as New Mer Kinase Inhibitors

Abnormal activation or overexpression of Mer receptor tyrosine kinase has been implicated in survival signaling and chemoresistance in many human cancers. Consequently, Mer is a promising novel cancer therapeutic target. A structure-based drug design approach using a pseudo-ring replacement strategy was developed and validated to discover a new family of pyridinepyrimidine analogs as potent Mer inhibitors. Through SAR studies, 10 (UNC2250) was identified as the lead compound for further investigation based on high selectivity against other kinases and good pharmacokinetic properties. When applied to live cells, 10 inhibited steady-state phosphorylation of endogenous Mer with an IC50 of 9.8 nM and blocked ligand-stimulated activation of a chimeric EGFR-Mer protein. Treatment with 10 also resulted in decreased colony-forming potential in rhabdoid and NSCLC tumor cells, thereby demonstrating functional anti-tumor activity. The results provide a rationale for further investigation of this compound for therapeutic application in patients with cancer

Discovery of Mer Specific Tyrosine Kinase Inhibitors for the Treatment and Prevention of Thrombosis

The role of Mer kinase in regulating the second phase of platelet activation generates an opportunity to use Mer inhibitors for preventing thrombosis with diminished likelihood for bleeding as compared to current therapies. Toward this end, we have discovered a novel, Mer kinase specific substituted-pyrimidine scaffold using a structure-based drug design and a pseudo-ring replacement strategy. The co-crystal structure of Mer with two compounds (7 & 22) possessing distinct activity have been determined. Subsequent SAR studies identified compound 23 (UNC2881) as a lead compound for in vivo evaluation. When applied to live cells, 23 inhibits steady-state Mer kinase phosphorylation with an IC50 value of 22 nM. Treatment with 23 is also sufficient to block EGF-mediated stimulation of a chimeric receptor containing the intracellular domain of Mer fused to the extracellular domain of EGFR. In addition, 23 potently inhibits collagen-induced platelet aggregation, suggesting that this class of inhibitors may have utility for prevention and/or treatment of pathologic thrombosis