Synthesis of carbon‐14 labeled 4‐[4‐[2‐[2‐[bis(4‐chlorophenyl) methoxy]ethyl sulfonyl][1‐14C]ethoxy]phenyl]‐1,1,1‐trifluoro‐2‐butanone

Carbon‐14 labeled 4‐[4‐[2‐[2‐[bis(4‐chlorophenyl)methoxyethylsulfonyl] [1‐¹⁴C]ethoxy]phenyl]‐1,1,1‐trifluoro‐2‐butanone was prepared in a six step radioactive synthesis from 2‐bromo[1‐¹⁴C]acetic acid. The overall radiochemical yield was 2.2%. The specific activity of the final product was found to be 42μCi/mg with a radiochemical purity of >98%. Copyright © 2002 John Wiley & Sons, Ltd.     

Synthesis of 1, 1′, 2, 2′, 3, 3′, 4, 4′ ‐ octadeutero‐sulforaphane

Sulforaphane (SFN), a naturally occurring isothiocyanate present in broccoli, shows strong evidence of anti‐carcinogenic activity. The mechanism of action, absorption, distribution, metabolism and excretion of the compound is however still poorly understood and requires a stable isotope labelled version of the compound for further studies. The paper describes an optimized synthesis of octadeutero‐SFN. Copyright © 2004 John Wiley & Sons, Ltd.     

New biological investigations on 3‐bromophenyl 6‐acetoxymethyl‐2‐oxo‐2H‐1‐benzopyran‐3‐carboxylate as anti‐angiogenic agent

The development of blood vessels inside tumors is required to provide the nutrients and oxygen needed for tumor growth and to allow the spread of cancer cells at a distance to form metastasis. Angiogenesis is also implicated in ocular diseases like age‐related macular degeneration. The present work describes the potential anti‐angiogenic properties of a coumarinic derivative, 3‐bromophenyl 6‐acetoxymethyl‐2‐oxo‐2H‐1‐benzopyran‐3‐carboxylate (IK9), previously described as a potent inhibitor of HT 1080 fibrosarcoma cell invasion in vitro and tumor growth in vivo. In vivo, ex vivo, and in vitro models were used to delineate the anti‐angiogenic properties of IK9. The anti‐angiogenic effect of IK9 was demonstrated in vivo in a choroidal neovascularization mice model and additionally ex vivo in a rat aortic ring assay where it was more active than the known matrix metalloproteinase inhibitor Ro 28‐2653. IK9 did not affect apoptosis, proliferation, or endothelial cell invasiveness in vitro. These findings suggest a complex mechanism of action of the compound via direct or indirect effects on endothelial cell properties. This study identifies IK9 as a new potent inhibitor of angiogenesis and suggests its potential use as a therapeutic agent. Drug Dev Res 71, 2010. © 2010 Wiley‐Liss, Inc.     

Synthesis of labelled N‐(4‐hydroxy‐[14C(U)]phenyl)‐2‐[2,3‐dihydro‐3‐oxo‐l,2‐benzisothiazol‐2‐yl‐1,1‐dioxide]acetamide

The amidation of 2‐[1,1‐dioxide‐3‐oxo‐1,2‐benzisothiazole‐2(3H)‐yl] acetyl chloride with carbon‐14‐labelled 4‐amino‐[¹⁴C(U)]phenol in NaOAc‐HOAc buffer solution at ?10°C gave N‐(4‐hydroxy‐[¹⁴C(U)]phenyl)‐2‐[2,3‐dihydro‐3‐oxo‐1,2‐benziso‐thiazol‐2‐yl‐1,1‐dioxide]acetamide in 82% yield. Subsequent hydrolysis with aqueous 0.5 N NaOH solution afforded the ring opened product N‐(4‐hydroxy‐[¹⁴C(U)]‐phenyl)‐2‐[2‐carboxy‐phenylsulfonamido]acetamide in 80% yield. Copyright © 2005 John Wiley & Sons, Ltd.     

Novel 1,3,5‐triazine derivatives exert potent anti‐cervical cancer effects by modulating Bax,Bcl2 and Caspases expression

This study aimed to develop novel 1,3,5‐triazine derivatives as potent anti‐cervical cancer agents. The compounds were synthesized in short steps with an excellent yield and characterized via various spectroscopic and analytical methods. A structure–activity relationship study suggested that electron‐withdrawing substituents showed greater anticancer activity than electron‐donating groups. Compound 7p (p‐fluoro) showed the highest activity against cervical cancer cells. In a nude mouse xenograft model inoculated with HeLa cells, 7p showed dose‐dependent inhibition of cervical tumour growth. Histopathological examination of excised tumour‐bearing tissues showed that 7p improved the microstructure in a dose‐dependent manner. Compound 7p also increased the proportions of HeLa cells in G0/G1 and S‐phase and significantly decreased that of G2/M‐phase. The effects of 7p on C‐caspase‐3, C‐caspase‐9, Bcl‐2 and Bax expression in HeLa cells were also determined.     

Synthesis and Evaluation of 2‐Alkylthio‐4‐(N‐substituted sulfonamide)pyrimidine Hydroxamic Acids as Anti‐myeloma Agents

A series of pyrimidine hydroxamic acids with a sulfide substituent at the second position and a sulfonamide substituent at the fourth position have been synthesized and evaluated for their activity against human myeloma cell line RPMI 8226. Several compounds exhibited significant anti‐cancer potency. It was found that representative compound 6a selectively killed cancerous but not normal cells. Moreover, compound 6a was effective in causing apoptosis in RPMI 8226 cells and exhibited promising HDAC‐inhibitory activities.     

From sentencing to execution – the processes of apoptosis

Objectives Cell proliferation and apoptosis play a major role in maintaining homeostasis and as such any disruption within these processes can lead to disease states. Apoptosis occurs in three non‐distinct phases – induction, effector and degradation – and can be executed through both the extrinsic and intrinsic pathways in addition to recognised sub‐pathways such as the p53 and lysosomal pathways. This review article highlights these pathways, incorporating an overview of the molecular regulators of apoptosis. Key findings These regulators include the prominent apoptotic players ‘the caspases’ in addition to the main regulators of the Bcl‐2 family. Increased understanding of the physiological processes of apoptosis at the molecular level not only offers an insight in disease pathogenesis but, in addition, allows for the development of diagnostic, prognostic and therapeutic tools. Summary While apoptosis remains the key player in cellular death, other processes cannot be dismissed. Many other proteins, in addition to caspases, within apoptotic pathways have been identified. Research continues into establishing the precise aspects of their molecular mechanisms of action and inter‐relationships. Inappropriate apoptosis due to dysregulation of cell death pathways provides a plethora of molecular checkpoints that can be targeted and modulated as part of therapeutic intervention. Increased research into these areas will prove useful for the design of novel chemotherapeutic drugs, an area that is particularly important due to increased risk of chemoresistance.     

甘草酸通过线粒体膜去极化诱导HPV18阳性的人宫颈癌HeLa细胞凋亡的作用研究

目的 分析甘草酸对人乳头瘤病毒（human papilloma virus,HPV）18阳性的人宫颈癌HeLa细胞的抗增殖和促凋亡特性及其可能的作用方式,探讨甘草酸在预防HPV感染的作用和治疗宫颈癌的潜力。方法 MTT法检测甘草酸在caspase-8,caspase-9,caspase-3抑制剂存在的情况下对HeLa细胞增殖的抑制作用,相差显微镜观察细胞形态学,4’,6-二脒基-2-苯基吲哚染色检测细胞核凝结和Hoechst 33342染色检测核形态,Annexin V-FITC/PI法分析细胞凋亡,采用caspase-8,caspase-9和caspase-3比色试剂盒测定宫颈癌细胞中caspase的活性,通过Mito Tracker Red CMX Ros染色评估线粒体膜电位（ΔΨm）。结果 与对照组相比,甘草酸可显著降低HeLa细胞活力,并伴随着核浓缩和DNA碎片的增加,呈剂量依赖性。甘草酸可通过线粒体去极化诱导宫颈癌HeLa细胞凋亡,并诱导caspase-8,caspase-9和caspase-3活化。结论 甘草酸对宫颈癌细胞的抗增殖和促凋亡特性可能是通过诱导线粒体膜电位的破坏,激活细胞内和细胞外途径的caspases活性,甘草酸可作为预防和控制宫颈癌的一种辅助手段。     

Synthesis and Antitubercular Activity of 2‐(substituted phenyl/benzyl‐amino)‐6‐(4‐chlorophenyl)‐5‐(methoxycarbonyl)‐4‐methyl‐3,6‐dihydropyrimidin‐1‐ium Chlorides

A series of 2‐(substituted phenyl/benzyl‐amino)‐6‐(4‐chlorophenyl)‐5‐(methoxycarbonyl)‐4‐methyl‐3,6‐dihydropyrimidin‐1‐ium chlorides 7–13 and 15 was synthesized in their hydrochloride salt form. The title compounds were characterized by FT‐IR, NMR (¹H and ¹³C) and elemental analysis. They were evaluated for their in vitro antitubercular activity against Mycobacterium tuberculosis H37Rv, multidrug resistance tuberculosis and extensively drug resistance tuberculosis by agar diffusion method and tested for the cytotoxic action on peripheral blood mononuclear cells by MTT assay. Among all the tested compounds in the series, compounds 7 and 11 emerged as promising antitubercular agents at 16 μg/mL against multidrug resistance tuberculosis and over 64 μg/mL against extensively drug resistance tuberculosis. The conformational features and supramolecular assembly of the promising compounds 7 and 11 were determined by single crystal X‐ray study.     

Facile synthesis of high specific activity 4‐[1‐14C]butyl‐1,2‐diphenylpyrazolidine‐3,5‐dione (phenylbutazone) using nucleophilic substitution

Metabolism, environmental fate, and low concentration food residue studies would be facilitated by the use of radiolabeled test articles that can be readily quantified within complex matrices. However, radiochemical approaches for such studies require high specific activities to allow analytical detection of correspondingly low masses of test article. The synthesis of high specific activity (>50 μCi/μmol) [¹⁴C]‐radiolabeled phenylbutazone presents a challenge using existing methodology, mainly due to the low solvent volumes required and vigorous refluxing needed to close the pyrazolidinedione ring. Herein, we report on the significant modification of an existing method that allows the synthesis of low masses of high specific activity (>50 μCi/μmol) [¹⁴C]‐phenylbutazone under mild conditions with simple purification and high yield. The closure of the pyrazolidinedione ring of 1,2‐diphenyl‐3,5‐pyrazolidinedione was accomplished as a first step with unlabeled 1,2‐diphenylhydrazine and diethyl malonate in 32% yield under gram‐scale conditions, which avoided the challenges of low solvent use and vigorous refluxing. Low mass of high specific activity n‐[1‐¹⁴C]‐butyl bromide was then added via a nucleophilic substitution reaction as a final step. Yields ranged from 65% to 92% during multiple synthetic attempts with unlabeled butyl bromide and were greatly influenced by reaction stoichiometry and the selection of base.     

Effect of non‐steroidal anti‐inflammatory drugs on non‐melanoma skin cancer incidence in the SKICAP‐AK trial

Recent studies link the prostaglandin metabolic pathway to skin carcinogenesis expanding possibilities that cyclooxygenase (COX) inhibitors may be utilized in non‐melanoma skin cancer (NMSC) chemoprevention. Using data from a study of the efficacy of retinol supplementation on incidence of NMSC, we sought to determine the role of non‐steroidal anti‐inflammatory drugs (NSAIDs) in NMSC development. Cox proportional hazards models describe the relationship between NSAID use and time to first squamous cell carcinoma (SCC) or basal cell carcinoma (BCC) among participants categorized by use pattern: continuous users (use for length of study duration), new users (use for less than study duration), and non‐users. For SCC and BCC, there was a statistically significant protective effect for participants who reported use for less than the study duration (HR = 0.49, 95%CI 0.28–0.87 and HR = 0.43, 95%CI 0.25–0.73, respectively). Categorical examination of NSAIDs (aspirin (ASA) vs. non‐ASA NSAIDs) showed significant effects for BCC among those using non‐ASA NSAIDs for less than the study duration (HR = 0.33, 95%CI 0.13–0.80). For SCC and BCC, NSAID use of shorter duration and potentially more recent, was more protective than longer duration of use. These results are counter to the idea that longer duration of NSAID use is more protective. Additional investigations are needed into the role NSAIDs play in the chemoprevention of NMSC. Copyright © 2009 John Wiley & Sons, Ltd.     

Synthesis of 14C‐labelled myosmine, [2′‐14C] ‐3‐(1‐pyrrolin‐2‐yl)pyridine

¹⁴C‐Labelled myosmine ([2′‐¹⁴C]‐3‐(1‐pyrrolin‐2‐yl)pyridine) was synthesized for autoradiography studies starting from [carboxyl‐¹⁴C]‐nicotinic acid by initial esterification of the latter in the presence of 1,1,1‐triethoxyethane. Without any purification the ethyl nicotinate formed was directly reacted with N‐vinyl‐2‐pyrrolidinone in the presence of sodium hydride, yielding ¹⁴C‐labelled myosmine. The product was purified by silica gel column chromatography. The radiochemical yield was 15% and the specific activity 55.2 mCi/mmol. Copyright © 2003 John Wiley & Sons, Ltd.     

Discovery of Novel PTP1B Inhibitors Derived from the BH3 Domain of Proapoptotic Bcl-2 Proteins with Antidiabetic Potency

BH3 peptide analogues are generally believed to exhibit great potency as cancer therapeutics via targeting antiapoptotic Bcl-2 proteins. Here, we describe the synthesis and identification of a new class of palmitoylated peptide BH3 analogues derived from the core region (h1–h4) of BH3 domains of proapoptotic Bcl-2 proteins and as alternative PTP1B inhibitors with antidiabetic potency in vitro and in vivo. PTP1B inhibitors are attractive for treatment of type 2 diabetes. We design the analogues using a simple lipidation approach and discovered novel lead analogues with promising antidiabetic potency in vitro and in vivo. The results presented here expanded the alternative target and function for the BH3 peptide analogues from one member Bim to other members of the proapoptotic Bcl-2 proteins and emphasize their therapeutic potential in T2DM. Furthermore, our findings may provide new proof of the regulatory function of Bcl-2 family proteins in mitochondrial nutrient and energy metabolism.     

Synthesis and pharmacological evaluation of 3‐(4‐chloro phenyl)‐2‐substituted‐3H‐quinazolin‐4‐ones as analgesic and anti‐inflammatory agents

A new series of 3‐(4‐chloro phenyl)‐2‐substituted‐3H‐quinazolin‐4‐ones were synthesized by reacting the amino group of 2‐hydrazino‐3‐(4‐chloro phenyl)‐3H‐quinazolin‐4‐one with different aldehydes and ketones. The compounds were investigated for their analgesic activity in albino mice, and for their anti‐inflammatory and ulcerogenic activities in Wistar rats. All test compounds exhibited analgesic and anti‐inflammatory activities. Compound VA2 (2‐(1‐ethylpropylidene‐hydrazino)‐3‐(4‐chloro phenyl)‐3H‐quinazolin‐4‐one) showed moderately more potent analgesic activity and compound VA3 (2‐(1‐methylbutylidene‐hydrazino)‐3‐(4‐chlorophenyl)‐3H‐quinazolin‐4‐one) showed moderately more potent anti‐inflammatory activity when compared with the reference standard, diclofenac sodium. The test compounds showed only mild ulcerogenic side effects when compared with aspirin. Drug Dev Res 69: 226–233, 2008 ©2008 Wiley‐Liss, Inc.     

Detection and metabolic investigations of a novel designer steroid: 3‐chloro‐17α‐methyl‐5α‐androstan‐17β‐ol

In 2012, seized capsules containing white powder were analyzed to show the presence of unknown steroid‐related compounds. Subsequent gas chromatography–mass spectrometry (GC‐MS) and nuclear magnetic resonance (NMR) investigations identified a mixture of 3α‐ and 3β‐ isomers of the novel compound; 3‐chloro‐17α‐methyl‐5α‐androstan‐17β‐ol. Synthesis of authentic reference materials followed by comparison of NMR, GC‐MS and gas chromatography‐tandem mass spectrometry (GC‐MS/MS) data confirmed the finding of a new ‘designer’ steroid. Furthermore, in vitro androgen bioassays showed potent activity highlighting the potential for doping using this steroid. Due to the potential toxicity of the halogenated steroid, in vitro metabolic investigations of 3α‐chloro‐17α‐methyl‐5α‐androstan‐17β‐ol using equine and human S9 liver fractions were performed. For equine, GC‐MS/MS analysis identified the diagnostic 3α‐chloro‐17α‐methyl‐5α‐androstane‐16α,17β‐diol metabolite. For human, the 17α‐methyl‐5α‐androstane‐3α,17β‐diol metabolite was found. Results from these studies were used to verify the ability of GC‐MS/MS precursor‐ion scanning techniques to support untargeted detection strategies for designer steroids in anti‐doping analyses. Copyright © 2015 John Wiley & Sons, Ltd.     

Investigation of anti‐inflammatory,nitric oxide donating,vasorelaxation and ulcerogenic activities of 1, 3‐diphenylprop‐2‐en‐1‐one derivatives in animal models

The main aim of this work is to find out novel chemical moieties with potent anti‐inflammatory and vasorelaxant activities with reduced gastric toxicities. For fulfilling the above aim, here we investigated novel chalcones (1, 3‐diphenylprop‐2‐en‐1‐one derivatives) with nitric oxide (NO) and hydrogen sulphide (H₂S) donating potency for anti‐inflammatory activity by carrageenan‐induced rat paw oedema. These molecules then further evaluated for in‐vitro NO‐releasing potency and vasorelaxation effect on isolated adult goat aortic tissue. The promising molecules were further screened for ulcerogenic activity in the rat model. The tested compounds produced % inhibition in paw oedema ranging from 29.16% to 79.69% and standard drug Diclofenac sodium produced 85.30% reduction in paw oedema after 5 hours. Out of this dataset, compounds AI1, AI7, Ca1, B2, B10, D2, and E8 showed 73.01%, 79.69%, 75.02%, 75.46%, 74.35%, 73.9% and 74.35% reduction in paw oedema respectively, which is approximately 80%–90% to that of standard Diclofenac sodium. The compound Ca1 was found to release 0.870 ± 0.025 mol/mol of NO and standard Glyceryl trinitrate (GTN) was found to release 0.983 ± 0.063 mol/mol of NO. The compound Ca1 produced 950.2 μmol/L of EC₅₀ whereas standard GTN produced 975.8 μmol/L of EC₅₀ for aortic smooth relaxation. The compounds Ca1 produced 0.1117 of ulcer index which is far less than that of standard Diclofenac sodium (1.148). The potent lead molecules were further evaluated to understand the mechanism of vasorelaxation by using specific antagonists or blockers of NO and H₂S.     

Microwave‐assisted synthesis of (RS) methyl‐2‐([2′‐14C]4,6‐dimethoxypyrimidin‐2′‐yloxy)‐2‐phenyl [1‐14C]ethanoate

We report here a facile synthesis of (RS) methyl‐2‐([2′‐¹⁴C]4,6‐dimethoxypyrimidin‐2′‐yloxy)‐2‐phenyl [1‐¹⁴C]ethanoate under microwave irradiation. Copyright © 2006 John Wiley & Sons, Ltd.     

Syntheses of [14C]‐labeled 2‐(3‐chlorophenyloxy)‐3‐[3‐(3‐hydroxy) pyridin‐4‐yl propoxy]pyridine,a phosphodiesterase 4 inhibitor and its metabolites

We describe here the synthesis of [¹⁴C]‐2‐(3‐chlorophenyloxy)‐3‐[3‐(3‐hydroxy)pyridin‐4‐yl propoxy]pyridine (1), a phosphodiesterase 4 inhibitor. [¹⁴C]‐Labeled 1 was prepared in three steps from [¹⁴C]‐2‐bromopyridin‐3‐ol in an overall yield of 32%. Preparation of [¹⁴C]‐labeled 2 and 3, two metabolites of 1, is also described. Copyright © 2014 John Wiley & Sons, Ltd.