Ketene dithioacetal mediated synthesis of 1,3,4,5-tetrasubstituted pyrazole derivatives and their biological evaluation

Abstract

Ketene dithioacetal mediated chemo- and regioselective synthesis of a series of novel 1,3,4,5-tetrasubstituted pyrazole derivatives (4a-l) integrated with a bioactive indole nucleus was achieved by reacting substituted 2-(1-methyl-1H-indole-3-carbonyl)-3,3-bis-(methylthio)-acrylonitrile (2) and substituted phenyl hydrazine hydrochloride (3) in the presence of a catalytic amount of anhydrous K₂CO₃ under reflux conditions. The structures were ascertained by ¹H NMR, NOESY, ¹³C NMR, FT-IR, and HRMS data. In vitro cytotoxicity evaluation of the synthesized compounds against MCF 7 (breast carcinoma) and normal Vero (monkey kidney) cell lines revealed that the compound 5-(5-Bromo-1-methyl-1H-indol-3-yl)-1-(4-cyano-phenyl)-3-methylsulfanyl-1H-pyrazole-4-carbonitrile (4k) showed significant cytotoxicity against MCF 7 (GI₅₀ = 15.6 µM) with low cytotoxicity against normal Vero cell line. Most of the synthesized compounds were also found to possess excellent anti-inflammatory and antioxidant (DPPH, NO, H₂O₂ and SOR) potential.     

Non-superimposable mirror image crystals of enantiomers by spontaneous resolution and the chiral discrimination mechanism

Non-superimposable mirror image crystals of both enantiomers(S/R) of cyclicγ-alkenyl alcohol(2) have been recognized and remarkably identified by the naked eye.More interestingly,both crystals are an outcome of most astonishingly H-bond and intermolecularσ/π-πinteractions.They accounted for the relatively rare and less predictable spontaneous resolution with optical purity >99%ee from the racemic mixture.The chiral discrimination mechanism of this spontaneous resolution has also been proposed.     

Photoactivatable Prodrugs of Highly Potent Duocarmycin Analogues for a Selective Cancer Therapy

A main problem of common cancer chemotherapy is the occurrence of severe side effects caused by insufficient selectivity of the applied drugs. A possible concept to overcome this limitation is light‐driven prodrug monotherapy. The synthesis as well as photochemical and biological evaluation of new photoactivatable prodrugs is described. Best results were obtained with prodrug (S,S)‐ 7 a . The photochemical labile protecting groups in (S,S)‐ 7 a can easily be removed by irradiation with UV‐A light in 30 min with a power of only 2 J cm^?2. The determination of the in vitro cytotoxicity by using an HTCFA‐test reveals a QIC₅₀ value of 8200 and the prodrug is more than two million times less cytotoxic than the corresponding seco‐drug (?)‐(S,S)‐ 5 with an IC₅₀ value of about 110 fM . The big therapeutic window makes (S,S)‐ 7 a very suitable for its use in selective cancer therapy.     

Screening active anti‐breast cancer compounds from Cortex Magnolia officinalis by 2D LC–MS

Most of the anti‐breast cancer drugs are often limited owing to drug resistance and serious adverse reactions. Therefore, development of more targeted and low toxic drugs from traditional Chinese medicines for breast cancer are needed. At the same time, establishment of fast and effective drug screening methods are urgently required. We describe here a 2D LC method of MDA‐MB‐231 cell membrane chromatography combined with HPLC/MS for recognition, separation, and identification of target components from traditional Chinese medicine Cortex Magnolia officinalis. The MDA‐MB‐231 cells membrane was used to prepare the chromatographic stationary phase in the first dimension. The active compounds had a retention characteristic on the cell membrane chromatography model (10 × 2.0 mm, 5 μm). The retention fractions were enriched using an online C₁₈ column (10 × 1.0 mm, 5 μm) and were analyzed by the second dimension RP chromatography. Finally, the activity of the retention fractions was tested through in vitro experiments. Results showed that the retention fractions were honokiol and magnolol and the inhibition rate on MDA‐MB‐231 cell growth were 23 and 64 μM, respectively. These results support the conclusion that this coupled analytical technique could be an efficient method in drug discovery.     

Periodic minimal surface organizations of the lipid bilayer at the lung surface and in cubic cytomembrane assemblies

The existence of infinite periodic lipid bilayer structures in biological systems was first demonstrated in cell membrane assemblies. Such periodicity is only possible in symmetric bilayers, and their occurrence is discussed here in relation to the asymmetry of cell membranes in vivo. A periodic membrane conformation in the prolamellar body of plants corresponds to a dormant state without photosynthesis. A similar reversible formation of a dormant state has also been observed in the mitochondria of the amoeba Chaos. In these cases the energy production has become insufficient to maintain the membrane asymmetry. Formation of membranes that are symmetric over the bilayer is proposed to be a principal mechanism behind formation of cubic membrane systems.     

A dual ligand targeted nanoprobe with high MRI sensitivity for diagnosis of breast cancer

Antibody targeted delivery is an effective strategy to improve the diagnostic imaging outcome of nanoscale imaging agents in the focal areas.Dual targeting micelles encapsulating superparamagnetic iron oxide were prepared from the amphiphilic block copolymer poly(ethylene glycol)-poly(ε-caprolactone)(PEG-b-PCL) with different targeting ligands cRGD and scFv-ErbB single chain antibody conjugated to the distal ends of PEG block.The breast cancer animal model was established by subcutaneous injecting the BT474 cells into the BALB/c-nu female nude mice and then employed to assess the potential of the dual ligand targeted magnetic micelles as a novel MRI contrast agent on a 1.5 T clinical MRI scanner.The T2 signal intensity of the tumor in animals receiving the dual ligand targeted magnetic micelles via tail vein decreased more significantly than the single ligand targeted and nontargeted magnetic micelles.These results indicate that the dual ligand targeted magnetic micelles,cRGD/scFv-ErbB-PEG-PCL-SPION,have great potential to act as a new type of effective nanoscale MRI contrast agent for early diagnosis of breast cancer.     

Design,Synthesis and Biological Evaluation of Novel Pyrazolo[1,2,4]triazolopyrimidine Derivatives as Potential Anticancer Agents

Three novel pyrazolo-[4,3-e][1,2,4]triazolopyrimidine derivatives (1, 2, and 3) were designed, synthesized, and evaluated for their in vitro biological activity. All three compounds exhibited different levels of cytotoxicity against cervical and breast cancer cell lines. However, compound 1 showed the best antiproliferative activity against all tested tumor cell lines, including HCC1937 and HeLa cells, which express high levels of wild-type epidermal growth factor receptor (EGFR). Western blot analyses demonstrated that compound 1 inhibited the activation of EGFR, protein kinase B (Akt), and extracellular signal-regulated kinase (Erk)1/2 in breast and cervical cancer cells at concentrations of 7 and 11 µM, respectively. The results from docking experiments with EGFR suggested the binding of compound 1 at the ATP binding site of EGFR. Furthermore, the crystal structure of compound 3 (7-(4-bromophenyl)-9-(pyridin-4-yl)-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine) was determined by single crystal X-ray analysis. Our work represents a promising starting point for the development of a new series of compounds targeting EGFR.     

Efficacy of a Covalent Microtubule Stabilizer in Taxane-Resistant Ovarian Cancer Models

Ovarian cancer often has a poor clinical prognosis because of late detection, frequently after metastatic progression, as well as acquired resistance to taxane-based therapy. Herein, we evaluate a novel class of covalent microtubule stabilizers, the C-22,23-epoxytaccalonolides, for their efficacy against taxane-resistant ovarian cancer models in vitro and in vivo. Taccalonolide AF, which covalently binds β-tubulin through its C-22,23-epoxide moiety, demonstrates efficacy against taxane-resistant models and shows superior persistence in clonogenic assays after drug washout due to irreversible target engagement. In vivo, intraperitoneal administration of taccalonolide AF demonstrated efficacy against the taxane-resistant NCI/ADR-RES ovarian cancer model both as a flank xenograft, as well as in a disseminated orthotopic disease model representing localized metastasis. Taccalonolide-treated animals had a significant decrease in micrometastasis of NCI/ADR-RES cells to the spleen, as detected by quantitative RT-PCR, without any evidence of systemic toxicity. Together, these findings demonstrate that taccalonolide AF retains efficacy in taxane-resistant ovarian cancer models in vitro and in vivo and that its irreversible mechanism of microtubule stabilization has the unique potential for intraperitoneal treatment of locally disseminated taxane-resistant disease, which represents a significant unmet clinical need in the treatment of ovarian cancer patients.