Publisher(s):China Academic Journals (CD Edition) Electronic Publishing House Co., Ltd.
ISBN:ISBN 978-7-499-00981-3 pdf
First Published: 2020.11.23
Discipline(s): Medicine & Public Health
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Tumor (Western Medicine), as part of China’s Medicine Progress Series, has 107 excellent articles on tumor treatment with western medicine. These articles were recommended by the editorial boards of the journals in which they were originally published in Chinese, covering comprehensive drug research, hematopoietic organ and lymphoid tumors, respiratory system tumor, nursing research, urogenital tumor, skin tumor, neurological tumor, and digestive system tumor. These articles fully demonstrate the cutting-edge advances in tumor treatment in China, with scientific methodological support and objective conclusions. The original articles were published in Chinese, and this book is a compilation of English version of selected articles.
CHENG Maosheng is currently serving as a director of Asian Federation for Medicinal Chemistry (AFMC), a director of Chinese Pharmaceutical Association, the vice chairman of the Specialized Committee on Medicinal Chemistry, Chinese Pharmaceutical Associati
1. Optimized expression, preparation of human papillomavirus 16 L2E7 fusion protein and its inhibitory effect on tumor growth in mice
Chinese Journal of Biotechnology,Part 1: Comprehensive Drug Research,Vol 31,No. 01
HPV16 L2E7 is a fusion protein used for therapeutical vaccine targeting HPV virus. To increase its expression in Escherichia coli, we optimized the codon usage of HPV16 l2e7 gene based on its codon usage bias. The optimized gene of HPV16 sl2e7 was cloned into three different vectors: p GEX-5X-1, pQE30, ET41a, and expressed in JM109, JM109(DE3) and BL21(DE3) lines separately. A high expression line was selected with pET41 a vector in BL21(DE3) cells. After optimization of the growth condition, including inoculation amount, IPTG concentration, induction time and temperature, the expression level of HPV16 L2E7 was increased from less than 10% to about 28% of total protein. HPV16 L2E7 protein was then purified from 15 L culture by means of SP Sepharose Fast Flow, Q Sepharose Fast Flow and Superdex 200 pg. After renaturing, HPV16 L2E7 protein with ≥ 95% purity was achieved, which was confirmed via SDS-PAGE gel and Western blotting. The combined use of purified HPV16 L2E7 and CpG helper has shown clear inhibition of tumor growth in mice injected with tumor cells, with six out of eight mice shown no sign of tumor. This study lays a solid foundation for a new pipeline of large-scale vaccine production.
2. Site-directed mutagenesis of human IL-29 and antineoplastic activity of the recombinant human IL-29 variant
Chinese Journal of Biotechnology,Part 1: Comprehensive Drug Research,Vol 31,No. 02
To explore the anti-tumor activity of human interleukin-29 (hIL-29) variant and based on bioinformatics analyzed data of hIL-29, a mutant gene hIL-29 mut33, 35 was amplified using site-directed mutagenesis and megaprimer PCR. The hIL-29 mut33, 35 was inserted into a eukaryotic expression plasmid pPIC9K and successfully expressed in Pichia pastoris GS115. A recombinant variant protein (rhIL-29 mut33, 35) was purified from the fermentation supernatant of the engineered GS115. To observe the antineoplastic activity of the variant rhIL-29 mut33, 35, a CCK-8 reagent was used to detect the anti-proliferative effect. Results showed that it had a strong anti-proliferative effect when acting on liver cancer cell BEL7402, colon cancer cell HCT8, and gastric cancer cell SGC7901. The inhibition ratios of the three tumor cells were (30.99 ± 1.58)%, (22.47 ± 1.37)% and (32.05 ± 2.02)%, respectively. In the high dose group, the anti-proliferative effect of the rhIL-29 mut33, 35 was stronger than that of wild type rhIL-29 (P < 0.01). This indicates the variant rhIL-29 mut33, 35 has potential development value for medicine.
3. Expression, purification of recombinant cationic peptide AIK in Escherichia coli and its antitumor activity
Chinese Journal of Biotechnology,Part 1: Comprehensive Drug Research,Vol 31,No. 03
AIK is a novel cationic peptide with potential antitumor activity. In order to construct the AIK expression vector by Gateway technology, and establish an optimal expression and purification method for recombinant AIK, a set of primers containing Att B sites were designed and used to create the Att B-TEV-FLAG-AIK fusion gene by overlapping PCR. The resulting fusion gene was cloned into the donor vector pDONR223 by att B and att P mediated recombination (BP reaction), then, transferred into the destination vector pDEST15 by att L and att R mediated recombination (LR reaction). All the cloning was verified by both colony PCR and DNA sequencing. The BL21 E. coli transformed by the GST-AIK expression plasmid was used to express the GST-AIK fusion protein with IPTG induction and the induction conditions were optimized. GST-AIK fusion protein was purified by glutathione magnetic beads, followed by rTEV cleavage to remove GST tag and MTS assay to test the growth inhibition activity of the recombinant AIK on human leukemia HL-60 cells. We found that a high level of soluble expression of GST-AIK protein (more than 30% out of the total bacterial proteins) was achieved upon 0.1 mmol/L ITPG induction for 4 h at 37 °C in the transformed BL21 E. coli with starting OD600 at 1.0. Through GST affinity purification and rTEV cleavage, the purity of the resulting recombinant AIK was greater than 95%. And the MTS assays on HL-60 cells confirmed that the recombinant AIK retains an antitumor activity at a level similar to the chemically synthesized AIK. Taken together, we have established a method for expression and purification of recombinant AIK with a potent activity against tumor cells, which will be beneficial for the large-scale production and application of recombinant AIK in the future.
4. Apoptosis induced by transferring cytosine deaminase suicide gene into kinds of cancer cells with novel polyamidoamine dendrimers
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 51,No. 04
This study demonstrates efficacy of a novel polyamidoamine dendrimers (PAMAM dendrimers) with pentaerythritol derivatives as the core (G5 PD dendrimer) in deliver of the cytosine deaminase ( CD) gene and EGFP gene fusion plasmid into different tumor cell lines to induce apoptosis. The physical and chemical properties of G5 PD dendrimers in terms of DNA complexation, particulate properties and transfection efficiencies were investigated and compared with commercial gene vectors PEI 25 kDa. The optimum ratio of G5 PD dendrimer complexed with plasmid DNA was 0.2:1, and the particle size of the complex was (100 ± 5) nm. Compared with the commercial gene carriers PEI, G5 PD dendrimer exhibited a higher transfection efficiency at the weight ratio of 1:1 in three different cell lines, given the fact that PEI are different from PAMAM dendrimers in terms of molecular structure. Furthermore, the cytotoxicity assays of the cell lines transfected with G5 PD dendrimer/pCD-EGFP-N1 followed by exposure to various concentrations of 5-fluorocytosine (5-FC) also showed that the transfected cell lines could generate a very low amount of 5-FC to 5-fluorouracil (5-FU) in a short period of time, which indicating the high expression level of CD gene in the cell line. These results indicate that the CD/5FC system of G5 PD dendrimer has an excellent efficacy in gene delivery.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 26,No. 05
Using pregnenolone as a starting material, α, β-unsaturated ketone was obtained by reacting with various aromatic aldehydes. Furthermore, compounds 4– 7 bearing a structure of pyrazoline were synthesized by the reaction of α, β-unsaturated ketone with hydrazine hydrate. The structures of compounds 4– 7 were characterized by NMR, IR and MS. The antiproliferative activity of target compounds was evaluated by MTT method against human colon carcinoma cells (HT-29), human cervical carcinoma cells (HeLa) and human gastric carcinoma cells (SGC-7901). The results showed that target compounds exhibited significant inhibitory activity to HeLa cells with IC 50 values of 7.7 μmol·L ?1, 8.5 μmol·L ?1, 7.4 μmol·L ?1, 9.1 μmol·L ?1(cisplatin: IC 50 = 10.1 μmol·L ?1), respectively.
Chinese Pharmaceutical Journal,Part 1: Comprehensive Drug Research,Vol 51,No. 06
Objective To observe the in vivo activity of Ranti-HER, a fully human monoclonal antibody, and combined with the doxorubicin or CPT-11 in established human tumor xenografts in nude mice, and to investigate whether EGFR expression is correlated with this activity. Methods The overall receptor of EGF was quantified by flow cytometry. The anti-tumor effects of Ranti-HER were evaluated using established, s/c human carcinoma xenografts in nude mice, and the relative growth rate of tumor was used to assess the anti-tumor activity. Results A431 cells showed highly expression of EGFR by flow cytometry, SW620 showed negative expression, and EGFR were expressed positively in HT29 and SW948 cells, but both of them were showed low expression. Ranti-HER (0.25–1.0 mg) could inhibit the tumor growth in human A431 epidermoid carcinoma xenografts and dose-effect relationship was observed; Ranti-HER (1.0 mg) could also inhibit the tumor growth in human SW948 colon carcinoma xenografts, but no anti-tumor effects of Ranti-HER 1.0 mg were observed in human HT29 and SW620 colon carcinoma xenografts. Therapeutic enhancement was observed in the A431 xenografts after treatment with Ranti-HER combined with doxorubicin. For another combination regimen, Ranti-HER and CPT-11 proved to be significantly more efficacious than Ranti-HER monotherpy in SW948 xenografts. Conclusion Antitumor activity of Ranti-HER are observed in xenografts in athymic nude mice, and the activity of Ranti-HER is correlated with the EGFR expression; synergistic effects are observed when Ranti-HER is combined with chemicals compared to Ranti-HER monotherapy.
Chinese Pharmaceutical Journal,Part 1: Comprehensive Drug Research,Vol 51,No. 07
OBJECTIVE To study the synthesis and antitumor activity of novel quinolone derivatives. METHODS Based on the structure of ciprofloxacin, the objective substances were designed and synthesized according to the principle of fragment-based drug discovery. Their antitumor activities in vitro were evaluated against A549, HL-60 and Hela cells by MTT assay. Molecular docking studies were performed with the Libdock protocol of Discovery Studio to afford the ideal interaction mode of the compound with the binding site of the Topo Ⅰ. RESULTS Eight novel compounds were synthesized and showed potential antitumor activities. CONCLUSION The antitumor activities of the synthesized quinolone derivatives are worthy of further study.
8. Synthesis and pharmacological evaluation of novel cyclopropane-containing semicarbazones derivatives as potential anticonvulsant and antitumor agents
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 26,No. 08
Epilepsy is a chronic brain disorder characterized by excessive temporary neuronal discharge and affects about 1% of the world’s population. The urgent demands for novel Antiepileptic drugs (AEDs) will be long-standing for the complexity and severity of epilepsy and the side effects of the currently marketed AEDs. Over last decade, anticonvulsant drug development has become one of the most active research areas in medicinal chemistry. According to combination principles, we found a way to mash up hydantoin core with semicarbazone. In other words, some atoms in those agents would be not only a part of the hydantoin core but also a member of the semicarbazone group. They would also be evaluated for their anticonvulsant activities by maximal electroshock shock (MES) and subcutaneous and pentylenetetrazole (scPTZ) seizure models, both of which are based on the tests of mice. Rotarod toxicity method was used to study the acute neurotoxicity profile of selected compounds. The preliminary anticancer activity was tested by the MTT assay. Nineteen novel target compounds Ia– Il and IIa– IIh were synthesized. The physiological activity of the new compounds is not due to cyclopropane structure. The activity experimental study showed that IIa– IId contained the lowest median effective dose (ED 50) of 100 mg·kg ?1 in MES test, and IIb– IId had the lowest ED 50 of 300 mg·kg ?1 in scPTZ test. In addition to compound III, VI and Ii– Ij, all compounds had severe neurotoxicity. In the mechanism of drug action and drug side effects, among these compounds, IC 50 value of compound IId against Hela was 23.7 μmol·L ?1 ; IC 50 values of compound Id against HepG2 was 26.7 μmol·L ?1. However, they did not show activities against the other cell lines which had been tested. These promising data suggested that the new compounds can be a new class of anticonvulsant and antitumor agents with high effectiveness and low toxicity for the treatment of epilepsy.
9. Design, synthesis and antiproliferative activity evaluation of EGFR inhibitors containing 3-hydroxy-4-pyridinone fragment
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 09
The development of multi-targeted tyrosine kinase inhibitors is of great significance for anti-cancer therapy. Epidermal growth factor receptor (EGFR) signaling pathway plays a crucial role in the apoptosis, proliferation, differentiation, migration and cell cycle of cancer cells. Meanwhile, Fe chelators are effective antitumor agents. By incorporating 3-hydroxy-4-pyridinone fragment (a Fe chelator motif) into the conventional quinazoline scaffold of EGFR inhibitors, 12 compounds which may target multiple drug targets were synthesized. The general procedure for the synthesis of target compounds was described and all of them were characterized by 1H-NMR, MS and melting point. In the tumor cell lines tested, most of the compounds exhibit efficient antiproliferative activity on the human epidermal carcinoma cell A431 (with EGFR overexpression) and HeLa, albeit a moderate inhibition in gefitinib-resistant NSCLC cell H1975 (bearing EGFR [L858R/T790M]).
10. Synthesis of binuclear copper(Ⅱ) complex with salicylaldehyde thiosemicarbazone: structure, anticancer properties and anticancer mechanism
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 10
One binuclear copper [Cu(II)]complex of the Schiff base ligand (salicylaldehyde thiosemicarbazone, HL), namely [(Cu II) 2(L) 2(H 2O)(MeOH)](NO 3 ?) 2, was synthesized and well characterized by single crystal X-ray diffraction, elemental analysis and infrared (IR) spectroscopic analysis. The Cu(II) complex strongly promoted the apoptosis of BEL-7404 cells and had a capacity to arrest the cell cycle at S phase of the cells. Furthermore, glutathione/oxidized glutathione (GSH/GSSG) assay, reactive oxygen species (ROS), mitochondrial membrane potential and western blot analyses revealed that the Cu(II) complex exerts its cytotoxicity through an ROS-mediated intrinsic mitochondrial pathway accompanied by the regulation of Bcl-2 family proteins.
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 52,No. 11
With the method of fluorescence polarization (FP), we screened small molecule inhibitors for PLK1 PBD to identify the lead compounds for antitumor drugs. FP led to the identification of a potent hit, F083-0063, whose inhibition rate was (99.7 ± 0.4)% at 10 μg·mL ?1. The IC 50 was calculated to be 1.9 ± 0.1 μmol·L ?1 using Graphpad Prism 5. The effect of the compound on cells' multiplication was measured by MTT assay which showed that F083-0063 inhibited the proliferation of many tumor cell lines. Flow cytometry analysis indicated that the F083-0063 promoted cell apoptosis and induced cell G 2/M arrest. Migration abilities of cells, evaluated using scratch test, increased significantly in the presence of F083-0063 with the migration rate as low as (37.6 ± 0.7)% at 20 μmol·L ?1. Molecular linkage technique found F083-0063 had good affinity with PLK1 PBD. The results of Western blotting showed that the expression of cyclin-dependent proteins was increased after treatment with F083-0063. In summary, F083-0063 has an antitumor activity and is expected to be an antitumor lead compound targeting PLK1 PBD.
Chinese Pharmacological Bulletin,Part 1: Comprehensive Drug Research,Vol 33,No. 12
Aim To explore whether the polypeptide vaccines CKL9 and YL20 can induce immune response and anti-tumor effect on HER-2 (+) tumors in vitro and in vivo, and to provide suggestions for clinical use. Methods The proliferation of specific lymphocytes and cytotoxic T lymphocyte activity (CTL) stimulated by CKL9 and YL20 were studied with CCK-8 assay and LDH assay, and the antitumor activity of CKL9 and YL20 was evaluated in vivo. Results The lymphocyte proliferation was promoted by incubation with CKL9 and YL20, and the relative increase of cells was 11.1% and 16.7% respectively at the concentration of 50 mg·L ?1 of CKL9 and YL20. The LDH assay confirmed the CTL effect induced by CKL9 and YL20 on HER2-positive tumor cells, not on HER2-negative tumor cells. With an effector–target ratio of 80:1, the inhibition of tumor cell by cytotoxic T-lymphocyte stimulated by CKL9 and YL20 could reach 89.8% and 84.3%, respectively. The HER2 (+) tumor cell N87 transplanted in Babes mice was inhibited by pre-immune polypeptide CKL9 and YL20. Conclusion The HER2-specific polypeptide vaccines CKL9 and YL20 could induce persistent specific CD4 and CD8 T cell immune and inhibit the growth of HER2 positive tumor cells.
13. In vitro metabolism and drug-drug interaction potential of IG-105, a novel antimicrotubule agent
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 52,No. 13
IG-105, N-(2,6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide, a novel antimicrotubule agent, showed potent anticancer activity in a variety of human tumor cells in vitro and in vivo. In order to characterize the metabolism and the possible drug-drug interaction of IG-105, we carried out a series of experiments. Drug metabolizing enzymes involved in IG-105 metabolism were investigated by using pooled human liver microsomes (HLMs) and recombinant cytochrome P450 isoforms (rP450s) respectively. The possible metabolites were analyzed by liquid chromatography-orbitrap-mass spectrometry (LC-Orbitrap-MS). The inhibitory effect of IG-105 on main P450 enzymes was also evaluated. The results showed that IG-105 can be metabolized by a series of rP450s, including CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4and CYP3A5, with the major contribution enzymes being CYP1A2, CYP2B6, CYP2C19, and CYP3A. Three metabolites (M1-M3) were identified and demethylation was the major phase I metabolic reaction for IG-105. IG-105 moderately inhibited CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A enzyme activities with IC 50 values of 6.4, 23.64, 0.39, 1.4 and 3.14 μmol·L ?1, respectively. Since the biotransformation of IG-105 involves multiple enzymatic pathways, the compound is less likely to be a victim of a concomitantly used medicine which inhibits activity of one of the CYPs. However, as IG-105 showed medium to strong inhibition on CYP1A2, CYP2D6, CYP3A and CYP2C19, caution is particularly needed when IG-105 is co-administrated with other anticancer drugs which are mainly metabolized by the above enzymes.
14. Preliminary study of lipid bilayer-coated calcium phosphate nanoparticles as a drug carrier for antitumor drug
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 52,No. 14
This study aims to prepare lipid bilayer-coated calcium phosphate core-shell nanoparticles (LCAPNs), which can dissolve in an acidic environment to improve the tumor cell toxicity of antitumor drug. Paclitaxel (PTX) loaded lipid coated calcium phosphate nanoparticles (PTX-LCAPNs) were prepared by thin-film dispersion method. The morphology, particle size and in vitro release behavior were characterized. Meanwhile, the intracellular uptake, intracellular dissolution, cell toxicity of PTX-LCAPNs and intracellular accumulation of PTX were evaluated in human HCC cell line (Huh-7). The results suggested that the mean diameter of the spherical LCAPNs was 124.73 ± 6.41 nm. The PTX-LCAPNs demonstrated little drug leakage in simulated normal physiological conditions, while a rapid release was observed in simulated intracellular condition in vitro. Moreover, the PTX-LCAPNs achieved 1.7 fold improvement in the intracellular PTX concentration leading to 5-fold reduction in half maximal inhibitory concentration (IC 50) values of PTX compared with calcium phosphate nanoparticles loaded with PTX (PTX-CAPNs), demonstrating a stronger cancer cell lethality.
15. Design, synthesis and antitumor activities of cyclopropane derivatives as novel B-Raf kinase inhibitors
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 15
Raf kinase, a common serine/threonine kinase, is part of Raf/M EK/ERK signal transduction pathway that plays a critical role in cell growth, differentiation and proliferation. It is also associated with the development and progression of tumor. B-Raf kinase has the strongest base activity in subtypes of three Raf kinases (A-Raf, B-Raf and C-Raf). Therefore, the development of B-Raf kinase inhibitors has been extensively studied in order to find some compounds as antineoplastic drugs. According to the mechanism, Raf kinase inhibitors can be divided into two categories, namely, typeⅠ and type Ⅱ. The former blocks the conduction of the signal by binding to the active conformation of the B-Raf kinase, which avoids the activation of the downstream MEK and ERK; the latter is bound to the inactive conformation of the B-Raf kinase, blocking the activation of the B-Raf kinase, resulting in direct inhibitory activity. Unfortunately, the anti-cancer activity produced by the current listing of Raf kinase inhibitors is mostly low. On the other hand, drug resistance, side effects, high price and other shortcomings limit their application. In this study, we designed 16 cyclopropane B-Raf kinase inhibitors, Ⅰ 1–Ⅰ 8 and Ⅱ 1–Ⅱ 8, by a series of structural optimization with sorafenib as the lead compound. Starting with p-hydroxybenzaldehyde, we conduct a series of key chemical reactions to get the target compounds, including the functional group protection, aldol condensation reaction, cyclization reaction, nucleophilic substitution reaction, deprotection, et al. Sixteen new compounds were synthesized, which were not reported in the literatures. The structures were identified by NMR. MTT assay was used to evaluate the effect of the target compounds on the anti-proliferation of Hep G2 cells. As a result, nine compounds showed good inhibitory activity in vitro, and the activity was better than that of the positive control. The results of structure-activity analysis showed that: increasing the volume of the substituent on pyridine amide may decrease the activity; the activity of the compounds with benzimidazole structure was higher than that with the acyl aromatic amine; the introduction of halogen or halogen-like groups, such as trifluoromethyl, may enhance the activity of the compounds.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 16
Anaplastic lymphoma kinase (ALK) and c-Met have been confirmed to play a decisive role in the process of various tumor cell migration, invasion and apoptosis and angiogenesis. ALK has been employed in the treatment of neuroblastoma (NB), one of the most common solid tumors in extra-crania frequently diagnosed in early childhood. And c-Met has also been considered to be a new target for the treatment of gastric carcinoma as it overexpressed significantly in gastric cancer cell strains. Crizotinib (PF-02341066) is a novel dual small-molecular ATP-competitive inhibitor of ALK and c-Met, whose structure is of great value for further research. Therefore, a series of novel compounds were designed and synthesized based on the structure of crizotinib and assessed their antiproliferative efficacy on human cancer cell strains. Furthermore, molecular docking simulation studies were performed using Discovery Studio 4.0 CDOCKER Docking software in order to obtain a deep sight into the binding mechanism of synthesized compounds ( 7a– 7h) with target proteins ALK and c-Met. These pyrazine derivatives containing 1,2,3-triazol were verified by 1H-NMR, 13C-NMR and MS spectral data. Their preliminary inhibition activities of tumor proliferation against human neuroblastoma (SH-SY5Y) and human gastric carcinoma (SNU-5) cell strain in vitro were tested by Alarm Blue assay. The results showed that some of the synthesized compounds displayed moderate to high levels of antitumor activities. Compound 7a showed best activity against SH-SY5Y cell strain (IC 50 = 0.467 μmol·L ?1) which was the same level as positive control crizotinib (IC 50 = 0.341 μmol·L ?1). As for inhibitory activity against SNU-5 cell strain, compound 7e showed the best activity with IC 50 value 148 nmol·L ?1, a promising value despite not better than crizotinib. In addition, the results of antitumor activities were basically in agreement with the Docking results evaluated by the CDOCKER score function.
17. Alkaloids from the Marine-Derived Fungus Aspergillus fumigatus YK-7 and Their Antitumor Activities
Chinese Pharmaceutical Journal,Part 1: Comprehensive Drug Research,Vol 52,No. 17
OBJECTIVE To study the secondary metabolites of marine-derived fungus Aspergillus fumigatus YK-7. METHODS The compounds were isolated by several column chromatographic techniques, including silica gel, ODS, Sephadex LH-20 column chromatography, and HPLC, and their structures were identified on the basis of physico-chemical properties and spectroscopic analysis. Trypan blue and MTT methods were applied for determining the effect of the compounds on proliferation of cancer cells in vitro. RESULTS Ten compounds were obtained, and their structures were identified as pseurotin A (1), pseurotin A 1 (2), 14-norpseurotin A (3), FD-838 (4), demethoxyfumitremorgin C (5), 9 β-hydroxyverruculogen TR-2 (6), 6-methoxyspirotryprostatin B (7), spiro [5 H, 10 H-dipyrrolo [1, 2-a:1', 2'-d] pyrazine-2- (3 H), 2'-[2 H] indole]-3', 5, 10 (1' H) -trione (8), terezine D (9), and 14-hydroxyterezine D (10). CONCLUSION Compounds 3, 6, 7, 9, and 10 are isolated from marine-derived fungus Aspergillus fumigatus for the first time. Compounds 1–4 exhibite moderate antiproliferative activity against selected cancer cell lines in vitro.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 18
An optimal process was developed after summarizing all the synthetic methods of brigatinib reported in literature and combining with a large number of experiments explored by the authors. Starting from 5-fluoro-2-nitroanisole, the key intermediate 5 was synthesized by substitution, reductive amination and reduction reaction. The key intermediate 9 was prepared starting from diethyl phosphite, followed by Grignard reaction, coupling and substitution reaction. The intermediate 5 was condensed with 9, followed by substitution to obtain brigatinib. This paper emphasized on optimizing the synthesis process of intermediates 4 and 7, which led to reducing the production cost greatly. The yield and purity of the target product were improved significantly. The total yield of brigatinib was 37.3% (calculated from diethyl phosphite) and its purity was 99.9% determined by HPLC. The structures of the target compound and some intermediates were characterized by 1H-NMR, 13C-NMR and MS.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 19
Thiazolidin-5-one derivatives have been reported to have various bioactivities, especially antitumor activity. On the basis of the previous researches, to design and synthesize imidazol-2-phenylamino-5-one derivatives, and evaluate their antitumor activity, with glycine as the starting material, the target compounds were synthesized by five steps, including cyclization, Knoevenagel condensation, N-alkylation, S-alkylation and substitution. The preliminary antitumor activities were tested by MTT method with cisplantin as the positive control. Eleven new compounds ( 1a– 1k) were synthesized, and their structures were identified by MS and 1H-NM R. The results revealed that compounds 1c and 1e showed high antitumor activity against HT-29, H460, A549 with IC 50 values of 0.38–1.44 μmol·L ?1 equivalent to that of cisplantin (IC 50 = 0.62, 0.22, and 0.35 μmol·L ?1). The preliminary study on structure-activity relationships has provided evidence for further structural transformation.
20. Preparation and evaluation of anti-tumor metastasis of self-assembly micelles loaded with indocyanine green in vitro
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 52,No. 20
In this study, we used low molecular weight heparin (LH) as hydrophilic sides, doxorubicin (DOX) as hydrophobic sides, and indocyanine green (ICG) as photosensitive drug to prepare pH-sensitive self-assembled polymeric micelles (LH-DOX/ICG). The micelles were prepared by dialysis, and evaluated for stability, pH sensibility in vitro, hemolytic test and photo-thermal effect. Wound healing test was used to evaluate the anti-metastatic effects. MTT assay and apoptosis detection kit were used to evaluate the cytotoxicity of micelles against melanoma B16 F10 cells. The size of the micelles was (148.7 ± 2.1) nm and the zeta potential was (?30.7 ± 1.1) mV. The drug-loading contents of DOX and ICG were 11.82% and 5.59%, respectively. The micelles exhibited spherical in shape, fairly uniform size. The micelles were stable within 48 h in 50% fetal bovine serum phosphate buffer. The release of DOX was pH-sensitive, while the release of ICG was sustained. The micelles were biocompatible and safe as indicated by the hemolytic test. The in vitro photo-thermal effect indicated LH-DOX/ICG micelles had similar photo-thermal effect to the free ICG. The wound healing test showed that the micelles had a good ability to inhibit tumor migration with laser irradiation. The MTT assay and cell apoptosis assay indicated that LH-DOX/ICG micelles displayed more efficient anti-tumor effect after near infrared laser compared to LH-DOX micelles. LH-DOX/ICG micelles are promising for the therapeutic effect of phototherapy and chemotherapy in combination for melanoma.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 27,No. 21
Histone deacelases (HDACs) play an important role in the promotion of malignant tumor cell development, survival and metastasis. HDACs have been identified as one of the most attractive target for new anticancer drug development. Based on the analysis of SAR of HDACs inhibitors, eight 1,4-dien-3-one benzamides HDACs kinase inhibitors were designed and synthesized. The target compounds were prepared through different methods with mild conditions and high yields. The structures of the target compounds were identified by MS and 1H-NM R spectra. The results of biological evaluation indicated that all compounds show ed high potency activities against A549, Hep3B, PANC-1 cancer cells, and further evaluation of these compounds are currently under investigation.
22. Construction of HER2-specific CAR-T cells and in vitro analysis of their activity to suppress tumor cell growth
Chinese Journal of Biotechnology,Part 1: Comprehensive Drug Research,Vol 34,No. 22
CAR-T cell therapy that targets surface antigens to kill tumor cells specifically has recently become another cornerstone in tumor immunotherapy. In this study, a lentiviral expression plasmid of CAR targeting human epidermal growth factor receptor 2 (HER2) was constructed by genetic engineering. The recombinant plasmid was co-transfected with other packaging plasmids into HEK293T cells by calcium phosphate precipitation to generate lenti-car, which are CAR lentiviral particles. HER2-specific CAR-T cells were obtained by transducing human peripheral blood mononuclear cells with lenti-car. Their specific inhibitory effects on HER2-positive and HER2-negative tumor cells were analyzed in vitro. The constructed CAR-T cells were specifically activated by HER2-expressing tumor cells as indicated by secretion of IFN-γ and IL-2. The inhibitory rate on HER2-positive SK-OV-3 cell line was (58.47 ± 1.72)%, significantly higher than that on the mock-treated control group ( P < 0.05). The inhibitory rate on HER2-negative K562 cell lines was (11.74 ± 2.37)%, which was not significantly different from that on the control group ( P > 0.05). Furthermore, when we transfected a HER2-expressing vector into K562, the inhibitory rate increased to (30.41 ± 7.59)%, which was higher than that on HER2-negative K562 ( P < 0.05). Thus, the constructed second-generation HER2-specific CAR-T cells specifically suppressed growth of tumor cells overexpressing HER2 protein, suggesting that HER2-specific CAR-T cells might prove useful for immunotherapy of HER2-positive cancer.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 23
In order to find the novel epipodophyllotoxins antitumor drugs and reduce toxicity and side effect, twenty novel 4'-demethylepipodophyllotoxin aromatic amide derivatives were designed and synthesized. Amino groups were introduced by Ritter reaction, and different side chains were introduced by condensation reaction. All of them were characterized by 1H-NMR and ESI-MS, and the effects of synthesized compounds against proliferations of A549, HepG2, KB, L1210 cells were evaluated by MTT test. The introduction of halogen in thiophene ring can increase the inhibition of A549, KB, L1210 cells; the introduction of halogen in indole ring can decrease the inhibition activity, however, the introduction of electron-donating group, such as methoxyl group can increase the inhibitory activity; carbon chain elongation can decease the inhibitory activity.
24. Isolation, purification and antitumor activity of bacillomycin D from Bacillus amyloliquefaciens Q-426
Chinese Journal of Biotechnology,Part 1: Comprehensive Drug Research,Vol 34,No. 24
Cyclic lipopeptide has extensive application prospect in the field of medicine due to its unique chemical structure and biological activity. This study aims to obtain high purity of cyclic lipopeptide monomer from Bacillus amyloliquefaciens strain Q-426, and illuminate preliminary antitumor mechanism of C-15 bacillomycin D and C-16 bacillomycin D. Firstly, crude cyclic lipopeptide solution was prepared by two-steps purification of acid precipitation and double-resins chromatography. In order to obtain purer product, preparative HPLC was utilized to separate and purify cyclic lipopeptide. Component 1 and component 2 were detected as C-15 bacillomycin D and C-16 bacillomycin D by HPLC-MS and ESI-MS/MS. Secondly, the effect of C-15 bacillomycin D, C-16 bacillomycin D, and their mixture (1:1, mol:mol) on cell proliferation was measured using human cancer cells (HeLa, MG, Hep-G2, and HT-29). The cyclic peptide showed a dose dependent manner on the cell proliferation inhibition of HeLa and MG cells. Finally, the results of the scratch wound healing assay and FACS analysis revealed that C-16 bacillomycin D could effectively influence the cell migration, and the cells treated with C-16 bacillomycin D showed typical apoptotic morphology. With the increase of drug concentration, the cells in the early apoptosis and late apoptosis increased, and G 0G 1 arrest was induced significantly.
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 53,No. 25
Due to the advantages of polymer micelles and the anticancer activity of doxorubicin (DOX), the polymer micelle of DOX is expected to be used for drug delivery in anticancer applications. As a biocompatible and biodegradable polymer, amphiphilic copolymer heparosan-adipic dihydrazide-vitamin E succinate (KV) can be self-assembled to form micelles with core-shell structure in aqueous phase. In this article, KV conjugates with two different degrees of substitution (DS) were synthesized to load DOX and were characterized by 1H NMR. The size distribution, morphology, zeta potential and release behavior in vitro of the DOX-loaded micelles were studied. In vitro cytotoxicity was investigated by MTT assay against MGC80-3 and COS7 cells. The cellular uptake of the DOX-loaded micelles was observed by fluorescence microscopy and flow cytometry. The 1H NMR spectra results confirmed the KV polymers were successfully conjugated and the degree of VES grafted on heparosan polysaccharide were 12% and 25%. Briefly, the micelles with two different DS were expressed as KV 12 and KV 25. The DOX-loaded micelles could resist serum adsorption because of the negative charge on the surface. The average particle size measured by dynamic light scattering (DLS) method was 140-150 nm and the TEM results indicated that the morphology of DOX-loaded micelles were spherical. The encapsulation efficiency and drug loading were 80% and 10%–15%, respectively. The DOX-loaded micelles had sustained release behavior and the cumulative release of DOX/KV 12 was slightly higher than DOX/KV 25. Moreover, the viabilities of cells which were co-incubated with blank micelles were greater than 90%. It is clear that the blank micelles almost non-toxic to both cells. The IC 50 of drug-loaded micelles against COS7 cells was much higher than that of MGC80-3 cells and theexhibited greater cytotoxicity. The cellular uptake of DOX/KV on MGC80-3 was greater than COS7 cells. In this study, KV polymer micelles have a sustained drug release activity and have a good selectivity to tumor cells, so it would be a potential carrier in drug delivery.
26. Impacts of particle shape on cellular uptake and tumor imaging application of ultra-pH sensitive micelles
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 53,No. 26
This study was designed to investigate the impacts of particle shape of micelles on cell uptake and tumor imaging applications. We designed and synthesized an ultra-pH sensitive PEG-iPDPA diblock polymer, and prepared it into wormlike micelles and spherical micelles by thin-film dispersion method and modified solvent evaporation method, respectively. Firstly, the pH responsiveness of two kinds of micelles was investigated in vitro. Both forms of micelles responded to pH sensitively, and each of them could reach 100 times of ON/OFF fold after conjugated with BDP fluorescent probe. Moreover, the cellular uptake of two forms of micelles depended on the concentration and incubation time. However, the amount of cellular uptake of spherical micelles was much higher than that of the wormlike micelles, which proved that the shape of nanoparticles had a great influence on the cellular uptake. The results of in vivo imaging revealed that the spherical micelles had a better tumor accumulation as well as tumor imaging outcomes. Finally, the biosafety of micelles was tested by MTT assay and H&E staining, which indicated that none of the two kinds of micelles had obvious toxicity. Collectively, these results suggest that the spherical micelles could be a better carrier compared with wormlike micelles in terms of cellular uptake, tumor accumulation and tumor detection.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 27
Patients with castration-resistant prostate cancer (CRPC) had limited therapeutic options once they became refractory to docetaxel chemotherapy, and no treatments improved survival until recently. In June 2010, the taxane anticancer drug cabazitaxel (Jevtana; Sanofi-Aventis), was approved in the US by FDA and in Europe by the EMA in combination with prednisone for the treatment of patients with CRPC whose disease progresses were after docetaxel treatment. As reported, more than 217 000 men have been diagnosed with prostate cancer and 32 000 will die of metastatic prostate cancer in the United States. Therefore, cabazitaxel will have the potential market prospect in future. Cabazitaxel is a semisynthetic taxane which possesses the same antitumor mechanism as paclitaxel and docetaxel. It is a 7,10- O-dimethylated derivative of docetaxel. The synthetic route of cabazitaxel is similar to that of docetaxel, but more difficult. Only few syntheses have been reported so far. Chattopadhyay S. K. et al. synthesized cabazitaxel in six steps with a total yield of 5.4% starting from 10-deacetylbaccatin Ⅲ (10-DAB) in 2002. In 2011, Sun et al. described dimethylation of the 7,10-OH of 10-DAB, and reaction with a protected (3 R, 4 S)- β-lactam, followed by deprotection, and gave cabazitaxel in 17.8% total yield. In 2012, Zhang et al. reported direct dimethylation of expensive docetaxel, which was prepared from 10-DAB in few steps, and could furnish cabazitaxel in 82% yield. Indeed, cabazitaxel could be synthesized by these methods. However, these methods, which normally needed column chromatography in purification procedure, are not suitable for scale-up. We found 2-(4-methoxy-phenyl)-4-phenyl-5-oxazoline acid ester is extremely unstable and easy to remove protection groups in acidic condition. Based on this finding, we designed a new synthetic method of cabazitaxel. The design involved seven steps and had the total yield of 41.5% with the purity of 99% started with available materials. The new synthesis, characterized by the simplicity of operation and the convenience of purification, is suitable for large-scale production.
28. Preparation of thermoresponsive micelles loaded with indocyanine green and doxorubicin for combined therapy in MCF-7 cells
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 53,No. 28
In this study, the thermoresponsive micelles were synthesized with random copolymerization method and the photosensitizer indocyanine green (ICG) was loaded on micelles through the physical adsorption. The light energy was converted into heat energy to increase the temperature after irradiation with near-infrared light. When the phase transition temperature was reached, the micelle was disassembled and the targeted therapy was achieved. The nanoparticles were characterized with a transmission electron microscopy, Fourier transform infrared spectrometer, nuclear magnetic resonance spectrometer and other characterization were used to investigate. The critical micelle concentration (CMC), upper critical solution temperature, the photothermal properties of the carrier and the release of drug triggered by light were investigated after the doxorubicin (DOX) loaded. The carrier was evaluated for toxicity, cellular uptake, the effect of photothermal, the combination of photothermal and chemotherapy; the p (AAm-co-AN)- g-PEG (PAAP) was spherical in shape with a particle size of about 45 nm and a phase transition temperature was about 43 ℃. The critical micelle concentration was 24 μg·mL ?1. The particle size increased to 88 nm after loaded with ICG and DOX which the photothermal effect was obvious. The cumulative release of the drug under the irradiation of near-infrared light (808 nm, 2 W·cm ?2, 2 min·h ?1) was increased to 59.4% (p H 5.0) after 5 h. The results of the cell experiment indicated that ICG-PAAP was almost non-toxic and uptaken by the lysosomal pathway. The cell killing effect was stronger with combination of chemotherapy (DOX as 20 μg·mL ?1) with more than 70% of the cells killed. The results showed that the prepared micelle with low toxicity was thermoresponsive and could be used in combined therapy of tumor under the irradiation of near-infrared light.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 29
This study focused on the synthesis and antitumor activity of the analogues of (3 S,4 S)-1-benzyl-4-phenyl-3-(pyridin-2-ylsulfonyl)piperidin-2-one and designed a series of δ-lactam compounds to evaluate their antitumor activity. Intermediate methyl 2-(pyridin-2-ylthio)acetate was synthesized by reacting pyridine-2-thiol with methyl bromoacetate, then was oxidized with potassium peroxymonosulfate to yield active methylene intermediate methyl 2-(pyridin-2-ylthio)acetate ( 3). The intermediate 3 reacted with different substituted cinnamaldehyde through Michael addition reaction with ( S)-2-(diphenyl((triethylsilyl)oxy)methyl)pyrrolidine as catalyst, then reacted with different substituted benzylamine through amination and cyclization reaction to provide target compounds. MTT method was used to evaluate the cytotoxicity of compounds on Karpass299 cells. In summary, thirty-nine chiral compounds were synthesized, and their antitumor activities on Karpass299 cells were explored. Among them, compound 6q exhibited good inhibitory activity (99% at 10 μmol·L ?1) on Karpass299 cells, demonstrating that it was a good new anti-lymphoma lead compound.
30. Synthesis and antitumor activity evaluation of quinoline derivatives containing α, β-enone fragments
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 30
Quinoline and chalcone derivatives have been reported to have various bioactivities, especially antitumor activity. In this study, a series of quinoline containing chalcone derivatives were designed and synthesized. The structures of the target compounds were identified by 1H-NMR, 13C-NMR and MS spectra. The antiproliferative activities in three human tumor cell lines were tested by MTT assay, and the topoisomerase inhibitory activities of target compounds were tested by topoisomerase mediated DNA relaxation assay. The results indicated that compound Ⅱ 1 displayed potent TopoⅡ inhibition activity at 50 μmol·L ?1. Most compounds showed strong cytotoxic activities against HepG2, A549, LNCaP cells. The structure–activity relationships studies indicated that changing the substituents dramatically impact the cytotoxic activities. The compounds showed the best cytotoxic activities when the fluorine substituent is introduced to the para-position of the benzene ring.
31. Design, synthesis and study of anti-tumor activity of thiazolinone urea derivatives bearing aryloxypyridinone fragments
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 53,No. 31
Taking cabozantinib as leading compound, 13 novel small-molecule c-Met inhibitors were designed and synthesized based on the obtained structure-activity relationships (SARs) of c-Met inhibitors. The structures of compounds were confirmed by 1H NMR, 13C NMR and HR-MS. In vitro anti-tumor activity was evaluated by MTT method, and the mechanism was preliminarily disclosed by real-time dynamic living cell imaging and flow cytometry analysis. The results indicated that most of compounds showed good inhibition activity against human non-small cell cancer cell A549 and human colorectal cancer cell HT-29 which was superior to cabozantinb. Compounds showed excellent cytotoxity and anti-proliferative activity against HT-29, and promoted cell apoptosis.
32. Synthesis and antitumor activity of arylaminopyrimidines bearing benzothiazine-1,1-dioxide moiety as ALK inhibitors
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 32
The rate of anaplastic lymphoma kinase (ALK) gene rearrangements in non-small cell lung cancer (NSCLC) tissues is 4%–6%. Aiming to explore novel anaplastic lymphoma kinase (ALK) inhibitors, a series of novel arylaminopyrimidine derivatives bearing benzothiazine-1,1-dioxide moiety were designed and synthesized based upon the cocrystal structure of ceritinib with ALKWT (PDB:4M KC) as well as the reported structure-activity relationships (SARs). Taking 2-fluoronitrobenzene as starting material, the critical intermediate 4-(2,5-dichloropyrimidin-4-yl)-3,4-dihydro-2 H-benzo[1,4- b]thiazine-1,1-dioxide was prepared through S-alkylation, chlorination, oxidation, reduction, intermolecular cyclization and N-alkylation reactions. Subsequently, ammonolysis reaction of 4-(2,5-dichloropyrimidin-4-yl)-3,4-dihydro-2 H-benzo[1,4- b]thiazine-1,1-dioxide with a variety of arylamine obtained newcompounds Ⅰ 1 - Ⅰ 11 . Compounds Ⅰ 12 – Ⅰ 15 were synthesized from compound Ⅰ 11 by Mannich reaction. The cellular and enzymatic assays validated compound Ⅰ 1 as a most promising ALK (IC 50 = 14.2 nmol·L ?1) inhibitor. Compound Ⅰ 1 showed significant inhibitory activities on ALK-dependent cell lines Karpas299 (IC 50 = 0.024 μmol·L ?1) and H2228 (IC 50 = 0.14 μmol·L ?1) superior to ceritinib (IC 50 = 0.048 μmol·L ?1 and 0.22 μmol·L ?1). The preliminary SARs studies suggested that the introduction of benzothiazine-1,1-dioxide moiety did improve the potency; diverse arylamine (R) and a variety of water-soluble groups (R 1) had dramatic impacts on cellular activity; overall, SARs identified molecules bearing the 2-methoxypyridine and 4-ethylpiperazine moieties with more favorable potency.
33. Design, synthesis and antitumor activity evaluation of novel steroidal spirooxindole derivatives based on pregnenolone
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 33
A series of novel steroidal spirooxindole-pyrrolidine derivatives were designed and synthesized. A series of 21-arylidenosteroids ( 3a– 3i) were prepared via the Claisen-Schmidt condensation reaction with readily available pregnenolone and aromatic aldehydes. Subsequently, the intermediates 3a– 3i reacted with isatin and sarcosine through 1,3-dipolar cycloaddition to afford the desired target compounds 6a– 6i. The structures of target compounds were characterized by 1H-NMR, 13C-NMR and HRMS. The target compounds were evaluated for antitumor activity against four selected human tumor cell lines of T24, SMM C-7721, MCF-7 and MGC-803. Several compounds exhibited moderate to potent activity. Particularly, compound 6a was more potent than 5-fluorouracil against T24, MCF-7 and MGC-803 cell lines with IC 50 values of 4.64, 8.82 and 7.74 μmol·L ?1, respectively. Compound 6d displayed the best antitumor activity against T24 than other human tumor cell lines.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 28,No. 34
Compound 990209, a novel kind of dithiocarbamate, has entered preclinical study as potent antitumor agent. However, the high dose and poor water solubility limited its further development. In order to increase the activity, six twin drugs of 990209 were designed and synthesized based on the principle of twin drugs design. Except for twin drug 3, all twin drugs were prepared with 990209 as the starting material. The base of 990209 was reacted with corresponding linkers affording twin drugs 1, 2 and 4. The N-chloroformylation intermediate ( 5 ' ) of 990209 was respectively reacted with ethane-1,2-diamine and 2- (aminooxy)- N, N-dimethylethan-1-amine to give the corresponding twin drugs 5 and 6. Twin drug 3 was synthesized from 1-Boc-piperazine through three steps. 1-Boc-piperazine was firstly reacted with linker affording the intermediate 3'. Then, the 3' was removed protecting group and reacted with CS 2 and 4-bromo-2, 2-diphenylbutanenitrile to afford the twin drug 3. All the target compounds were confirmed by NMR and HR-MS and evaluated for their antitumor activity against Bel-7402 by MTT assay. It was found that the twin drug 6 was ten-fold potent than alone compound 990209. This result showed that the linker of the twin drug 6 was suitable and the twin drug 6 was worthy of further study.无
35. Synthesis and antitumor activity of fluoroquinolone C-3 arylidene thiazolone derivatives from ciprofloxacin
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 54,No. 35
To expand an efficient strategy for the conversion of antibacterial activity of fluoroquinolones into an antitumor activity, sixteen new compounds, 1-cyclopropyl-6-fluoro-7-(4-methyl-piperazin-1-yl)-3-(5-arylidenethiazol-4(5 H)-one-2-yl)-quinolon-4(1 H)-ones ( 7a–7p), were designed and synthesized with a thiazolone ring and an arylidene moiety as an isostere and modified group, respectively, from ciprofloxacin. Their structures were characterized by elemental analysis and spectral data. The in vitro antitumor activities of the synthesized compounds were measured using Hep-3B, Capan-1 and HL60 cell lines and were found to be more potent than ciprofloxacin. Meanwhile, the SAR revealed that the halogenated phenyl compounds such as fluorophenyl ( 7h, 7i), chlorophenyl ( 7j, 7k) or bromophenyl compounds ( 7l, 7m), and aromatic heterocyclic substitution such as furyl ( 6n) or pyridyl compounds ( 6o, 6p) displayed better activities than the control compounds, especially the IC 50 values of pyridyl compounds 6o and 6p against Capan-1 cell growth were comparable to doxorubicin. Thus, an arylidene-modified thiazolone scaffold as the replacement of the C-3 carboxylic acid group appears to be an alternative route for an improved antitumor activities of fluoroquinolones.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 29,No. 36
Poly(ADP-ribose) polymerase-1 (PARP-1) is involved in the process of DNA repair and mediates the integrity of genomic function. By catalyzing the transfer of ADP-ribose from nicotinamide adenine dinucleotide (NAD +) to its receptor proteins, PARP-1 participated in the base excision repair pathway of single-stranded damaged DNA. For homologous recombination repair (HRR)-deficient cancer cells like breast cancer 1/2 (BRCA1/2), inhibition of PARP-1 makes it unable to perform effective reconstitution of damaged DNA fragments. With the approval of olaparib, rucaparib, and niraparib, PARP-1 has become a promising target for cancer therapy. Most of PARP-1 inhibitors have an amide structure in their pharmacophore mimicking the structure of the nicotinamide moiety of NAD +. Herein, two series of compounds bearing 6,8-difluoro-2-thioxo-2,3-dihydroquinazolin-4(1 H)-one and 6-fluoro-2-thioxo-2,3-dihydropyrido[2,3- d]pyrimidin-4(1 H)-one scaffolds were designed, synthesized and evaluated for their effects on cell proliferation in human cancer cell line MDA-MB-436 by Alarm Blue assay. Preliminary results of proliferation inhibition activity against MDA-MB-436 cell lines in vitro indicated that the target compounds have certain antitumor activity, with IC 50 values ranging from 14.1 to 1 246 nmol·L ?1. The inhibitory activity of compounds 16a– 16e against MDA-MB-436 cells tended to be more potent than that of compounds 16f– 16j. Among all designed compounds, compound 16c exhibited slightly lower inhibitory potency than the positive control olaparib, with IC 50 values 14.1 nmol·L ?1 and 4.2 nmol·L ?1, respectively.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 29,No. 37
Isocitrate dehydrogenase (IDH), an important metabolic regulator in vivo, has attracted much attention due to its specific functional mutation and high mutant frequency in many human tumors. The excessive accumulation of 2-HG after IDH mutation can competitively inhibit a series of alpha-KG-dependent dioxygenases, resulting in hypermethylation of histone and DNA. This leads to abnormal epigenetic regulation and thereby promotes the occurrence and development of tumors. Therefore, mutant IDH (mIDH) has become a new target for anti-cancer therapy. Based on the existing mIDH2 inhibitor AGI-6780, we designed the target compounds I and II by using the ring-fusing strategy and bioisosterism. The target compounds were synthesized from o-bromonitrobenzene by sulfonation, nucleophilic substitution, reduction, nucleophilic substitution with 2-chloro-4-trifluoromethylbenzimidazole or urea formation. The inhibitory activities of the target compounds against IDH2 R140Q were determined by an enzymatic assay. The results showed that most of the compounds displayed good inhibitory activities. Among them, compounds I-1 and II-1 exhibited the most potent activity, which deserved further investigation.
38. Construction and anti-tumor efficacy of a peptide vaccine that targets calcium-binding protein S100A9
Acta Pharmaceutica Sinica,Part 1: Comprehensive Drug Research,Vol 54,No. 38
Calcium-binding protein S100A9 is closely related to inflammation and tumor invasion, and is one of the specific markers of myeloid-derived suppressor cells (MDSCs). In this study, a recombinant polypeptide vaccine CTB-S100A9 targeting mouse calcium-binding protein S100A9 was constructed by fusion cholera toxin B subunit (CTB) with S100A9 gene. The CTB-S100A9 fusion protein was expressed in E coli. and purified by Ni + affinity chromatography. Vaccinate the purified recombinant CTB-S100A9 protein supplemented with aluminum hydroxide adjuvant can break the autoimmune tolerance and produce high titer of S100A9 antibody in mice. Moreover, the S100A9 antibody produced by CTB-S100A9 vaccination is more specific and does not cross-react with S100A8. In the mouse 4T1 breast cancer model, CTB-S100A9 vaccination not only has significant tumor prevention effects, but also has significant tumor therapeutic effects. In addition, CTB-S100A9 significantly inhibited lung metastasis in 4T1 mice breast cancer model. Further analysis by flow cytometry showed that CTB-S100A9 vaccination can significantly reduce the tumor induced Treg cells and granulocyte-derived MDSC in 4T1 mice model, and reverse the tumor immunosuppressive environment, thereby promote the anti-tumor efficacy. The animal experiments in this study were carried out under the animal care guidelines approved by the Animal Ethics Committee of the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. This study shows that CTB-S100A9 is a good recombinant vaccine that targets the tumor immunosuppression environment and has great potential for the future clinical application.
Chinese Pharmacological Bulletin,Part 1: Comprehensive Drug Research,Vol 35,No. 39
Aim To investigate the role of gap junction (GJ) composed of Cx26, Cx32 and Cx43 respectively on the anti-tumor effect of miR-124 and the underlying mechanisms. Methods Hela cells, stably transfected by PBI plasmid coding for Cx32 or Cx26 (called Hela32 or Hela26 cells), and U87 cells, transfected with shRNA-Cx43 (called U87 shRNA-Cx43) were cultured in vitro. Connexin expression was examined by Western blot, and GJ function was estimated by parachute assay. The impact of miR-124 on cell proliferation was detected by standard colony forming assay, and the transfer of Cy3-labelled miR-124 through GJ channel was observed by patch clamp. Results Cx32 or Cx26 expression and GJ function were induced by doxycycline (Dox, the promotor for PBI plasmid) in transfected Hela cells. MiR-124 reduced the proliferation of Hela cells, Dox incubation alone did not affect Hela cell growth, and also had no effect on anti-tumor effect of miR-124 when combined with miR-124 transfection. Compared with U87 shRNA-NC, the Cx43 expression and GJ function significantly decreased in U87 shRNA-Cx43. Similar to the effect on Hela cells, MiR-124 also reduced U87 cell growth. Reducing Cx43 expression did not influence U87 cell proliferation, but attenuated the growth-inhibition effect of miR-124 when combined with miR-124 transfection. Under the microscope, the transfer of fluorescence-labelled miR-124 from “donor” cell to adjacent “non-injection” cell was observed. Conclusions The role of GJ on anti-tumor effect of miR-124 possesses connexin heterogeneity. Compare with Cx26 or Cx32, GJ composed of Cx43 has more obvious effect, which may be related to the maximum permeability of junction channel to miR-124.
Chinese Journal of Medicinal Chemistry,Part 1: Comprehensive Drug Research,Vol 29,No. 40
Programmed cell death 1 (PD-1) is an important immune checkpoint, which can induce apoptosis of antigen-specific T cells and reduce apoptosis in regulatory T cells. Programmed cell death ligand 1 (PD-L1) is the critical ligand of PD-1, extensively expressed on the surface of marrow-derived cells, non-lymphoid tissues, solid organ, and tumor cell. The block of PD-L1 expressed by host cell can promote tumor recession. Although anti-PD-1/PD-L1 antibodies have been approved for the treatment of multiple types of cancer, they may produce side effects because of their immunogenicity. Small-molecule inhibitors of the PD-1/PD-L1 interaction can overcome the shortcomings. Bristol-Myers Squibb reported the small molecule inhibitors of PD-1/PD-L1, which can induce PD-L1 dimerization. Based on the reported PD-L1 inhibitor BMS-1018, ten biphenyl derivatives were designed and synthesized by swapping the position of the substitutes, bioisosterism, and molecular docking technology. The structures of the target compounds were confirmed by 1H-NMR and ESI-MS. The inhibitory activities of these compounds against PD-L1 were determined by the homogeneous time-resolved fluorescence (HTRF) binding assay. The results demonstrated that the target compounds showed varying degrees of inhibitory effects on PD-L1. Among them, A-1, A-3, A-4, and C-2 displayed the most potent activity, suggesting that these compounds might be the candidates for further investigation.