New medium oxacyclic O,N-acetals and related open analogues: biological activities

Attention is increasingly being focussed on the cell cycle and apoptosis as potential targets for therapeutic intervention in cancer. Taking 1-[(2-oxepanyl)]-5-fluorouracil previously prepared by us, we committed ourselves to increase the lipophilicity of this upper cyclohomologue of Ftorafur and prepared a series of bioisosteric benzannelated seven-membered 5-FU O,N-acetals to test them against the MCF-7 human breast cancer cell line. Benzo-fused seven-membered O,O-acetals or their acyclic analogues led to the expected 5-FU O,N-acetals (or aminals), in addition to six- and to 14-membered aminal structures and acyclic compounds. All the cyclic aminals provoked a G(o)/G(1)-phase cell cycle arrest, whereas Ftorafur, a known prodrug of 5-FU, and 1-[2-(2-hydroxymethyl-4-nitrophenoxy)-1-methoxyethyl]-5-fluorouracil (51) induced an S-phase cell cycle arrest. Although breast cancer is most often treated with conventional cytotoxic agents it has proved difficult to induce apoptosis in breast cancer cells and, consequently, improved clinical responses may be obtained by identifying therapies that are particularly effective in activating apoptosis. 1-(2,3-Dihydrobenzoxepin-2-yl)-5-fluorouracil (26) may be particularly useful in stimulating apoptosis in breast cancer. This compound is more potent as an apoptotic inductor than paclitaxel (Taxol). Finally, a fact that is worth emphasizing is that the cyclic and acyclic 5-FU O,N-acetals induce neither toxicity nor death in mice after one month's treatment when administered intravenously twice a week, with a 50 mg/kg dose each time. Taken together, the experimental findings provide evidence of specific anti-tumour activity of these new substances and warrant further evaluation in in vivo models of breast cancer to future clinical applications.     

Antitumoural properties of benzannelated seven-membered 5-fluorouracil derivatives and related open analogues. Molecular markers for apoptosis and cell cycle dysregulation

Attention is increasingly being focussed on the cell cycle and apoptosis as potential targets for therapeutic intervention in cancer. We prepared a series of bioisosteric benzannelated seven-membered 5-FU O,N-acetals to test them against the MCF-7 human breast cancer cell line. Benzo-fused seven-membered O,O-acetals or their acyclic analogues led to the expected 5-FU O,N-acetals (or aminals), in addition to six- and 14-membered aminal structures and acyclic compounds. All the cyclic aminals provoked a G0/G1-phase cell cycle arrest, whereas Ftorafur, a known prodrug of 5-FU, and 1-[2-(2-hydroxymethyl-4-nitrophenoxy)-1-methoxyethyl]-5-fluorouracil (11) induced an S-phase cell cycle arrest. Although breast cancer is most often treated with conventional cytotoxic agents it has proved difficult to induce apoptosis in breast cancer cells, but improved clinical responses may be obtained by identifying therapies that are particularly effective in activating apoptosis. 1-(2,3-Dihydrobenzoxepin-2-yl)-5-fluorouracil (5) may be particularly useful in stimulating apoptosis in breast cancer.     

Studies on antitumor agents, 2. Syntheses and antitumor activities of 1-(tetrahydro-2-furanyl)-5-fluorouracil and 1,3-bis(tetrahydro-2-furanyl)-5-fluorouracil.

Convenient and efficient methods were developed for preparing 1-(tetrahydro-2-furanyl)-5-fluorouracil (Thf-FU, 3) [trade name, Futraful (Ftorafur) or FT-207], which is used clinically as an antitumor agent, and 1,3-bis(tetrahydro-2-furanyl)-5-fluorouracil (Thf2-FU, 4). For the syntheses, 2,4-bis(trimethylsily)-5-fluorouracil (Me3Si-FU, 1) and 2-acetoxytetrahydrofuran (Thf-OAc, 2) were condensed in the presence of Friedel-Crafts catalysts, such as SnCl4 and BF3-Et2O in dichloromethane, or in the presence of NaI in acetonitrile to give Thf-Fu or Thf2-FU depending on the reaction conditions and workup procedure. A trace of 3-(tetrahydro-2-furanyl)-5-fluorouracil (3-Thf-FU, 5) was formed in these reactions. Thf2-FU was easily hydrolyzed to Thf-FU. 2-Methoxytetrahydrofuran can be used instead of Thf-OAc for preparation of Thf-FU under similar conditions. The optimal ratios of Me3Si-FU, Thf-OAc, and SnCl4 or NaI for preparation of Thf-FU and Thf2-FU were determined. In all cases, 2-2.5 equiv of Thf-OAc with respect to Me3Si-FU gave the best results. The yields of Thf-FU and more especially of Thf2-FU were greatly dependent on the relative amount of SnCl4, and 0.01-0.1 equiv of the catalyst with respect to Me3Si-FU gave the best results. Thf2-FU was found to be effective against murine solid tumors and it was less toxic than Thf-FU when given orally. The antitumor activity of 3-Thf-FU is also reported.     

Novel derivatives of 5-fluorouridine and 5-fluorouracil having potent antitumor and lower immunosuppressive activities.

We studied the biological activities of several 5-fluorouridine (5-FUR) and 5-fluorouracil (5-FU) derivatives to find novel antitumor drugs with lower immunosuppressive effects. We examined 5-FUR and 5-FU derivatives acylated with (2-n-propyl-n-pentanoyl)glycine (KN-539). Among the examined compounds, we found satisfactory activities in a derivative of 5-FUR, 2',3',5'-tris-O-[N-(2-n-propyl-n-pentanoyl)glycyl]-5-fluorouridine (UK-21), and a derivative of 5-FU, 1-(6-[N-(2-n-propyl-n-pentanoyl)glycyl] amino-n-hexylcarbamoyl)-5-fluorouracil (UK-25). UK-21 (0.05-0.2 mmole/kg, p.o., 10 days) and UK-25 (0.1-0.4 mmole/kg, p.o., 10 days) suppressed Meth A and E.L.4 tumor growths in the corresponding syngeneic hosts (BALB/c mice and C57BL/6 mice, respectively) without decreasing body weight and blood leukocyte count. UK-21 and UK-25 suppressed the proliferation of KB tumor cells in vitro (IC50: 3.0 x 10(-11) M and 4.4 x 10(-7) M, respectively) at concentrations almost equivalent to those of 5-FUR and 5-FU, respectively. These results suggest that UK-21 and UK-25 express their antitumor activity as 5-FUR and 5-FU, respectively. Neither UK-21 nor UK-25 suppressed thymus weight and humoral antibody production against sheep red blood cells (SRBC) in ddY mice, although 1-(2-tetrahydrofuryl)-5-fluorouracil (FT-207) and 5-FU suppressed them in their respective therapeutic dose ranges for tumors. Thus, UK-21 and UK-25 are expected to develop into anticancer drugs with lower immunotoxicological effects.     

5-氟尿嘧啶衍生物的合成及其体外抗肿瘤活性研究

目的设计合成5-氟尿嘧啶衍生物,并对其抗肿瘤活性进行评价。方法以5-氟尿嘧啶(5-FU)结构为基础,化学合成2-苄氧基-5-氟-4(3H)-嘧啶酮(2-BF),采用质谱(MS)、核磁共振氢谱(1H-NMR)及碳谱(13C-NMR)对其结构进行表征;MTT比色分析法比较2-BF与5-FU作用于人结肠癌细胞(SW620)及正常人肠上皮细胞(HIEC)后对细胞生长抑制率的影响及差异。结果 MS、1H-NMR和13C-NMR的结果确证合成化合物为目标产物;体外实验结果表明,2-BF(0.01～100μmol·L-1)作用于SW620细胞24h和48h后,其对细胞的生长抑制率分别为29.20%～64.96%、32.85%～72.26%,呈浓度、时间依赖性。在HIEC细胞中,高浓度2-BF(1、10、100μmol·L-1)的细胞生长抑制作用明显低于5-FU(P<0.01)。结论本实验成功合成了5-氟尿嘧啶衍生物——2-苄氧基-5-氟-4(3H)-嘧啶酮,不仅具有较好的抗肿瘤活性,且细胞毒性明显低于5-FU,具有潜在的临床应用价值。     

3-全乙酰化半乳吡喃糖基-5-氟尿嘧啶的合成及其体外抗肿瘤活性研究

目的设计合成5-氟尿嘧啶半乳糖衍生物,并对其抗肿瘤活性进行评价。方法以5-氟尿嘧啶结构为基础,化学合成3-全乙酰化半乳吡喃糖基-5-氟尿嘧啶(3-PGF),采用质谱(MS)及核磁共振氢谱(1H-NMR)对其结构进行表征;MTT比色分析法比较3-PGF与5-FU作用于人结肠癌细胞(SW-1116)及正常人肠上皮细胞(HIEC)后,其对细胞生长抑制率的差异。结果 MS和1H-NMR的结果确证合成化合物为目标产物;体外实验结果表明,3-PGF(0.01～100μmol.L-1)作用于SW-1116细胞48 h后,其对细胞的生长抑制率为8.45%～65.53%,呈浓度依赖性。在HIEC细胞中,3-PGF的细胞生长抑制作用明显低于5-FU。结论本实验成功合成了5-氟尿嘧啶半乳糖衍生物——3-全乙酰化半乳吡喃糖基-5-氟尿嘧啶,不仅具有较好的抗肿瘤活性,同时,该衍生物的毒性明显低于5-FU,为5-FU衍生物的设计合成提供了新的思路。     

Antitumor activity of 2-amino-4,4alpha-dihydro-4alpha, 7-dimethyl-3H-phenoxazine-3-one against Meth A tumor transplanted into BALB/c mice

We examined the in vivo effect of 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3- one (Phx) on Meth A carcinoma cells transplanted into BALB/c mice, in terms of both antitumor activity and side effects. Phx, which was synthesized by the reaction of 2-amino-5-methylphenol with bovine hemolysates, was administered i.p. at doses of 1 and 5 mg/kg to BALB/c mice transplanted with Meth A tumor cells. Phx exerted a strong antitumor activity to Meth A tumor growing in the mice as 5-fluorouracil (5-FU) did. The antitumor activity of Phx at the dose of 5 mg/kg was comparable to that of 5-FU at the dose of 7.8 mg/kg. In contrast, unlike 5-FU, Phx did not cause leukopenia while showing a strong antitumor activity. The compound also produced little changes in body weight and no wasting of mice developed. These results show that Phx has strong anti-tumor activity, but exerts lower side effects and suggest that Phx is available for therapeutic purposes in the future.     

Induction of cytokines and killer cell activities by cisplatin and 5-fluorouracil in head and neck cancer patients

It has been suggested that certain antitumor agents stimulate antitumor immunity. In the present study, we examined whether cisplatin and 5-fluorouracil (5-FU) accelerate the antitumor host responses in head and neck cancer patients. Two groups of patients were studied, i.e. an untreated (UT) group and a treated, disease-free (TDF) group that received chemo-immunotherapy in combination with radiotherapy and operation. When peripheral blood mononuclear cells (PBMC) derived from head and neck cancer patients were treated with cisplatin or with 5-FU, interferon-gamma, tumor necrosis factor (TNF)-alpha, TNF-beta, interleukin (IL)-1beta, IL-6, IL-12 and IL-18 as well as killer cell activities were significantly induced in both groups. In this case, these activities induced by cisplatin in UT showed lower levels than those in TDF, whereas the activities induced by 5-FU in the UT group demonstrated almost similar levels to those in TDF. These activities were significantly inhibited by anti-asialo-GM1 antibody. Furthermore, cytokine levels in sera and killer activities of PBMC derived from the cancer patients were significantly increased after cisplatin administration. These findings suggest that cisplatin and 5-FU increase anticancer immunity mediated by induction of cytokines and killer cell activities in patients with head and neck cancer.     

2-(芳基哌嗪)乙酰氨基-5-取代-苯并噻唑衍生物的合成及抗肿瘤活性

目的合成新型苯并噻唑衍生物并研究其抗肿瘤活性。方法以3-取代苯胺为原料合成系列2-（芳基哌嗪）乙酰氨基-5-取代-苯并噻唑衍生物,采用MTT法测试了化合物对肿瘤细胞的抑制作用。结果合成了12个新的苯并噻唑衍生物,化合物结构经’H-NMR、ESI-MS和元素分析确证。结论多数目标化合物对5种肿瘤细胞株具抗增殖作用,部分化合物显示出与阳性对照药物5-氟尿嘧啶相当的抗肿瘤活性。     

Studies of antitumor agents. 1. Resolution of racemic 1-(tetrahydro-2-furanyl)-k-fluorouracil into the R and S isomers and examination of the biological activities of the isomers.

1-(Tetrahydro-2-furanyl)-5-fluorouracil (Thf-FU), which is named Ftorafur or FT-207 and is used clinically as an antitumor agent, was conveniently synthesized by condensation of the trimethylsilyl derivative of 5-fluorouracil with 2-acetoxytetrahydrofuran using NaI as a catalyst. This optically inactive Thf-FU was resolved into optically active (R)-(+)- and (S)-(-)-Thf-FU in high optical purity and excellent yield by formation of diastereoisomers with brucine. 13C NMR data were obtained on Thf-FU and related compounds and the antibacterial activities and in vivo antitumor activities of these isomers were tested. The degradations of these isomers to 5-fluorouracil by liver microsomes were also examined. No significant differences were found in any of these properties of these isomers.     

复方氟尿嘧啶多相脂质体对S180荷瘤小鼠肿瘤生长抑制及其不良反应

目的:比较复方氟尿嘧啶多相脂质体(Co-5-FU)和氟尿嘧啶(5-FU)对小鼠肉瘤细胞系S180荷瘤小鼠的抗肿瘤作用及不良反应。方法:将小鼠肉瘤S180细胞悬液接种于小鼠背部皮下或腹腔,建立S180荷瘤小鼠实体瘤和腹水瘤模型。取70只小鼠,分成实验组(Co-5-FU)、阳性对照组(5-FU)和正常对照组,分别腹腔注射40 mg·kg~(-1)、20 mg·kg~(-1)、10 mg·kg~(-1)3种不同浓度的Co-5-FU和5-FU,正常对照组腹腔注射生理盐水。比较Co-5-FU和5-FU对实体瘤小鼠的肿瘤抑制率,腹水瘤小鼠的生命延长率,同时通过血生化和病理等检测观察2种药物对各脏器的不良反应。结果:与正常对照组比较,40 mg·kg~(-1)、20 mg·kg~(-1)、10 mg·kg~(-1) Co-5-FU对S180腹水瘤小鼠的生命延长率分别是52．7％,41．4％和31．9％(P＜0．05),对S180实体瘤小鼠的肿瘤抑制率分别是48．8％和39．2％和32．0％(P＜0．01);同等剂量Co-5-FU比5- FU引起的骨髓抑制轻(P＜0．05);40 mg·kg~(-1)5-FU可引起肝脏ALT、AST、GGP升高(P＜0．05)。结论: Co-5-FU比同等剂量的5-FU具有更强的抗肿瘤作用,对心脏、肾脏、肝脏、骨髓的不良反应明显小于5- FU。     

A protein-bound polysaccharide immunomodulator, PSK, does not suppress the conversion from 1-(2-tetrahydrofuryl)-5-fluorouracil to 5-fluorouracil in patients with gastric cancer.

Effects of the immunomodulator PSK on the metabolism of 1-(2-tetrahydrofuryl)-5-fluorouracil (tegafur) to 5-fluorouracil (5-FU) were examined in 10 patients with advanced gastric cancer and who had undergone curative resection. PSK is a protein-bound preparation, extracted from Coriolus versicolor and belongs to Basidiomycetes. The 5-FU concentration in the plasma was 0.024 micrograms/ml at 15 min after the intravenous injection of 400 mg of tegafur and the area under the curve of 5-FU was 0.58 micrograms.h/ml. Following administration of PSK, 3 g/day for 8-14 months, there was no change in the plasma level of 5-FU, in any patient. As the clinical dose of PSK had no apparent influence on the metabolism of tegafur to 5-FU, the combination of PSK and tegafur can be prescribed to treat patients with advanced gastric cancer.     

Synthesis and cytotoxic effects of deoxy-tomentellin

Cannabigerol (1, CBG), methyl 4-[(2E)-3,7-dimethyl-2,6-octadienyl)oxy]-3-methoxybenzoate (2, DTM), 5-fluorouracil (3, FU) as a reference, and cannabidiol (4, CBD) were tested for their growth inhibitory effects against KB(ATCC NO, OCL 17) cell lines using two different assays, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay and the sulforhod-amine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of IC50 values of these compounds in tumor cell lines showed that their susceptibility to these compounds decreases in the following order: DTM > CBD > 5-FU > CBG by MTT assay and DTM = CBD > 5-FU > CBG by SRB assay. CBG 1, DTM 2, 5-FU 3, and CBD 4 were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different assays, the MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of CD50 values of these compounds in NIH 3T3 fibroblasts shows that their susceptibility to these compounds in decreases the following order(:) CBD > 5-FU > DTM > CBG by MTT assay, CBD > 5-FU > CBG > DTM by SRB assay. These results suggest that DTM 2 has the most growth-inhibitory activity against KB cell lines.     

N1-(芳)烷酰氧亚甲基-5-氟尿嘧啶的合成及其初步抗肿瘤活性的研究

目的　合成抗肿瘤活性高 ,毒性小的N1- (芳 )烷酰氧亚甲基取代的 5 -氟尿嘧啶衍生物。方法　以 5 -氟尿嘧啶为原料 ,经与甲醛加成反应后 ,和二酸单苄酯反应即得目标物 3a、3b、3c ,并采用MTT法及SRB法评价目标物 3的抗肿瘤活性。结果　合成了 3个目标物 3a、3b、3c ,其结构经1HNMR、IR和MS确证。结论　体外抗肿瘤活性筛选结果显示 3b和 3c有较强的抗肿瘤活性。     

含1，3，4-噻二唑的5-氟尿嘧啶衍生物的合成及其抗肿瘤活性

目的 制备低毒、高抗瘤活性的N<,1>-乙酰氨基-(5-烃基/芳基-1,3,4-噻二唑-2-基)-5-氟尿嘧啶衍生物.方法 以5-氟尿嘧啶为原料,在氢氧化钾的作用下与氯乙酸反应后,与合成的中间体2-氨基-5-取代-1,3,4-噻二唑反应制得目标物,并评价其抗肿瘤活性.结果 和结论 合成了7个未见文献报道的目标物3 a～3 g,并用<'1>HNMR、HRMS、IR确认了结构;目标化合物3 d、3 f和3 g有较好的抗肿瘤活性.     

杂氮硅三环与5-氟尿嘧啶结合物的合成

杂氮硅三环具有免疫增强和抗肿瘤作用，依据药物设计的拼合原理设计了杂氮硅三环与5-氟尿嘧啶结合的协同前药，以期寻找高效低毒的抗肿瘤化合物。合成了2个未见报道的杂氮硅三环与5-氟尿嘧啶的结合物。其结构经质谱、核磁共振氢谱及元素分析确证。     

Ringtransformation von 2-Phenacylidenoxazolidinen: Untersuchungen zur Regiochemie der Addition von Hydroxylamin und zur Ringöffnung spiroanellierter Intermediate

Ring Transformation of 2-Phenacylidenoxazolidines: Investigations Concerning the Regiochemistry of the Addition of Hydroxylamine and the Ring Opening of Spiroannelated Intermediates Acylketene O,N-acetals having the structure of 2-phenacyliden-oxazolidines 2 are formed by reaction of lithiated 2-alkyl-4,5-dihydrooxazoles 1 with benzoic acid esters. Under the conditions used, competitive 1,2- and 1,4-addition is observed. The ring opening of the resulting spiroannelated intermediates ( 3 and 4 , resp.) occurs regiospecifically at the O-atom, leading to 5-(β-hydroxyethyl)-aminoisoxazoles 5 and 3-(β-hydroxyethyl)-aminoisoxazoles 7 , respectively. Possible reaction mechanism are discussed, the structures of the regioisomeric compounds are substantiated by spectrometric methods, especially ms and ¹³C-NMR investigations.     

葡萄糖苯丙苷类化合物的合成及其抗肿瘤活性

目的设计合成葡萄糖苯丙苷衍生物，并寻求具有抗肿瘤活性的新化合物。方法四乙酰基溴代葡萄糖与醇经过成苷、脱保护、与苯甲醛缩合3步反应得到目标化合物。以A431（人表皮鳞癌细胞）、A549（人肺腺癌细胞）、7721（人肝癌细胞）3种肿瘤细胞为测试细胞株，采用M1vr法评价了目标化合物的抗肿瘤活性。结果与结论合成了19个糖苷衍生物，其结构均经。H—NMR确证。体外抗肿瘤活性实验表明，4，6-O-亚苄基β－D－吡喃葡萄糖－3－（4－甲氧基苯基）丙苷（1j）显示出较好的抗肿瘤活性。     

Sulfonyl-containing nucleoside phosphotriesters and phosphoramidates as novel anticancer prodrugs of 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP)

A series of sulfonyl-containing 5-fluoro-2'-deoxyuridine (FdU) phosphotriester and phosphoramidate analogues were designed and synthesized as anticancer prodrugs of FdUMP. Stability studies have demonstrated that these compounds underwent pH dependent beta-elimination to liberate the corresponding nucleotide species with half-lives in the range of 0.33-12.23 h under model physiological conditions in 0.1 M phosphate buffer at pH 7.4 and 37 degrees C. Acceleration of the elimination was observed in the presence of human plasma. Compounds with an FdUMP moiety (4-9) were considerably more potent than those without (1-3) as well as 5-fluorouracil (5-FU) against Chinese hamster lung fibroblasts (V-79 cells) in vitro. Addition of thymidine (10 microM) reversed the growth inhibition activities of only 5-FU and the compounds with an FdUMP moiety, but had no effect on those without. These results are consistent with thymidylate synthase as the target of the prodrugs.