Long-circulating and liver-targeted nanoassemblies of cyclic phosphoryl N-dodecanoyl gemcitabine for the treatment of hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a serious cancer with high mortality worldwide. Gemcitabine (GEM) is easily degraded in the circulation and has no tumor-targeted effect. In our previous research, an amphiphilic GEM derivative, cyclic phosphoryl N-dodecanoyl gemcitabine (CPDG) was prepared based on the techniques of HepDirect prodrug and self-assembled drug delivery systems (SADDS), which self-assembled into the stable nanoassemblies in water. In this study, the long-circulating nanoassemblies of CPDG/CHS-PEG1500 (9:1, mol/mol) were prepared for HCC treatment. In vitro and in vivo studies of the long-circulating CPDG nanoassemblies were explored. The degradation rates of CPDG depended on the media. CPDG showed much faster degradation in the acidic environment (pH 2.0) than the weak acidic and neutral media (pH 5.0, pH 7.4). However, the degradation half-life (t_1/2) of CPDG was about 43 h in the mouse plasma, longer than the t_1/2 at pH 2.0. Therefore, the long-circulating CPDG nanoassemblies could keep stable before reaching the targets in vivo. In the biodistribution study, the long-circulating CPDG nanoassemblies were bolus intravenously (i.v.) injected into the hepatocellular tumor-bearing mice. The distribution of CPDG in the tumors was much higher than that in the blood, indicating the tumor targeting of the long-circulating nanoassemblies. In the pharmacodynamic study, the long-circulating CPDG nanoassemblies were i.v. injected into the tumor-bearing mice with doses of (37.5, 75 μmol/kg) compared with GEM (150 μmol/kg). The mice were injected once every 3 days for totally 3 times. The long-circulating nanoassemblies nearly always showed the higher anti-cancer effects than GEM. The tumor inhibitory rates of GEM, the long circulating CPDG nanoassemblies (37.5, 75 μmol/kg) were 49.54, 42.97, 65.10%, respectively. Therefore, the long-circulating CPDG nanoassemblies had the much higher anti-cancer effect than GEM. The long-circulating CPDG nanoassemblies are promising nanomedicines to treat HCC. The combination design of tumor-targeted nanoassemblies based on HepDirect prodrug technique and SADDS theory is an effective method to modify the pharmacologically active nucleosides to treat some liver diseases.     

Design,synthesis, and biological evaluation of target water‐soluble hydroxamic acid‐based HDACi derivatives as prodrugs

Four compounds T1 , T2 , T3 , and T4 were designed and synthesized as Vorinostat and Belinostat derivatives being the target water‐soluble prodrugs. The water solubility of Vorinostat derivatives, T1 and T2 , exhibited 400‐ to 600‐fold higher than that of Vorinostat, and Belinostat derivatives, T3 and T4 , showed 600‐ to 750‐fold higher than that of Belinostat. Four compounds were evaluated for their inhibitory activities against tumor cell lines HT‐29 and Hut‐78 in the absence or presence of β‐D‐glucuronidase. The inhibitory effects of T1 and T2 were comparable to Vorinostat in the presence of β‐D‐glucuronidase, but were higher than 10 μM in the absence of β‐D‐glucuronidase. Therefore, T1 and T2 are promising candidates for in vivo investigations with high potential to be the target water‐soluble prodrugs. IC₅₀ values of Belinostat derivatives T3 and T4 were not affected by β‐D‐glucuronidase, but T3 and T4 had the excellent cell proliferation inhibition on Hut‐78.     

两种化疗方案辅助治疗乳腺浸润性导管癌的成本-效果分析

目的：评价注射用表柔比星（法玛新）联合注射用多西他赛注射液（艾素）和注射用紫杉醇脂质体（力朴素）治疗乳腺浸润性导管癌药物成本-效果。方法：回顾某院2016-2017两年期间收治的经组织病理学确诊的乳腺癌患者病历,从中筛选符合标准的患者,使用注射用表柔比星（法玛新）联合多西他赛注射液（艾素）方案的患者为ED组,使用注射用表柔比星（法玛新）联合注射用紫杉醇脂质体（力朴素）方案的患者为EP组,分析比较2组的成本-效果。结果：2组总有效率为74.29%和79.41%,差异无统计学意义（P>0.05）;2组均有不同程度的不良反应,发生率分别为91.43%、73.53%,差异有统计学意义（P<0.05）。ED与EP组的成本-效果比分别为930.70、1218.15,ICER=538 895.27（>3倍人均GDP）,ED组具有成本-效果优势。结论：表柔比星联合多西他赛（艾素）与联合紫杉醇脂质体（力朴素）方案治疗浸润性乳腺癌效果相当,但ED组较为经济。     

紫杉醇热敏脂质体的制备与质量控制

目的：制备紫杉醇热敏脂质体,建立测定其含量、有关物质及溶血磷脂的方法,考察其含量、有关物质、溶血磷脂量以及体外溶血性。方法采用高效液相色谱（ HPLC）-紫外法检测制剂含量及有关物质,HPLC-电雾式检测器检测制剂中溶血磷脂单棕榈酰磷脂酰胆碱的量,并对检测方法进行验证;分光光度法测定体外溶血百分率。结果紫杉醇在60.39~181.17μg·mL-1范围内峰面积与浓度线性关系良好,回收率及精密度实验均符合要求;有关物质测定方法专属性、灵敏度和系统适用性均符合要求;溶血磷脂测定方法的专属性和灵敏度符合有关规定,在1.5~50.0μg·mL-1范围内峰面积与浓度线性关系良好,回收率、重复性均符合要求;3批自制紫杉醇热敏脂质体的含量在90.0%~110.0%之间,单个杂质含量均〈0.5%,总杂质含量均〈2.0%,体外基本不引起溶血。结论含量、有关物质及溶血磷脂检测方法均可用于紫杉醇热敏脂质体的质量评价,自制紫杉醇热敏脂质体各项指标均符合要求,且质量稳定。     

The flavoprotein FOXRED2 reductively activates nitro-chloromethylbenzindolines and other hypoxia-targeting prodrugs

The nitro-chloromethylbenzindoline prodrug SN29428 has been rationally designed to target tumour hypoxia. SN29428 is metabolised to a DNA minor groove alkylator via oxygen-sensitive reductive activation initiated by unknown one-electron reductases. The present study sought to identify reductases capable of activating SN29428 in tumours. Expression of candidate reductases in cell lines was modulated using forced expression and, for P450 (cytochrome) oxidoreductase (POR), by zinc finger nuclease-mediated gene knockout. Affymetrix microarray mRNA expression of flavoreductases was correlated with SN29428 activation in a panel of 23 cancer cell lines. Reductive activation and cytotoxicity of prodrugs were measured using mass spectrometry and antiproliferative assays, respectively. SN29428 activation under hypoxia was strongly attenuated by the pan-flavoprotein inhibitor diphenyliodonium, but less so by knockout of POR suggesting other flavoreductases contribute. Forced expression of 5-methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR), as well as POR, increased activation of SN29428 in hypoxic HCT 116 cells. SN29428 activation strongly correlated with expression of POR and also FAD-dependent oxidoreductase domain containing 2 (FOXRED2), in cancer cell lines. This association persisted after removing the effect of POR enzyme activity using first-order partial correlation. Forced expression of FOXRED2 increased SN29428 activation and cytotoxicity in hypoxic HEK293 cells and also increased activation of hypoxia-targeted prodrugs PR-104A, tirapazamine and SN30000, and increased cytotoxicity of the clinical-stage prodrug TH-302. Thus this study has identified three flavoreductases capable of enzymatically activating SN29428, one of which (FOXRED2) has not previously been implicated in xenobiotic metabolism. These results will inform future development of biomarkers predictive of SN29428 sensitivity.     

Cannabidiol inhibits paclitaxel-induced neuropathic pain through 5-HT1A receptors without diminishing nervous system function or chemotherapy efficacy

Background and Purpose

Paclitaxel (PAC) is associated with chemotherapy-induced neuropathic pain (CIPN) that can lead to the cessation of treatment in cancer patients even in the absence of alternate therapies. We previously reported that chronic administration of the non-psychoactive cannabinoid cannabidiol (CBD) prevents PAC-induced mechanical and thermal sensitivity in mice. Hence, we sought to determine receptor mechanisms by which CBD inhibits CIPN and whether CBD negatively effects nervous system function or chemotherapy efficacy.

Experimental Approach

The ability of acute CBD pretreatment to prevent PAC-induced mechanical sensitivity was assessed, as was the effect of CBD on place conditioning and on an operant-conditioned learning and memory task. The potential interaction of CBD and PAC on breast cancer cell viability was determined using the MTT assay.

Key Results

PAC-induced mechanical sensitivity was prevented by administration of CBD (2.5 – 10 mg·kg⁻¹) in female C57Bl/6 mice. This effect was reversed by co-administration of the 5-HT_1A antagonist WAY 100635, but not the CB₁ antagonist SR141716 or the CB₂ antagonist SR144528. CBD produced no conditioned rewarding effects and did not affect conditioned learning and memory. Also, CBD + PAC combinations produce additive to synergistic inhibition of breast cancer cell viability.

Conclusions and Implications

Our data suggest that CBD is protective against PAC-induced neurotoxicity mediated in part by the 5-HT_1A receptor system. Furthermore, CBD treatment was devoid of conditioned rewarding effects or cognitive impairment and did not attenuate PAC-induced inhibition of breast cancer cell viability. Hence, adjunct treatment with CBD during PAC chemotherapy may be safe and effective in the prevention or attenuation of CIPN.     

多西他赛和紫杉醇致严重不良反应对比分析

目的：分析多西他赛和紫杉醇致严重不良反应的特点,为临床安全用药提供参考。方法收集2004年1月至2013年1月我省药品不良反应监测中心收到的多西他赛和紫杉醇引起的严重不良反应报告,比较2种药物所致严重不良反应临床表现及转归。结果共收集多西他赛和紫杉醇严重不良反应报告31份,涉及患者31例,不良反应33例次。31例患者中男性9例,女性22例;年龄31～76岁,平均年龄53岁。紫杉醇引起的严重不良反应17例次,主要临床表现为过敏性休克（70.59%）、过敏样反应（11.76%）、白细胞减少（11.76%）、骨髓抑制（5.88%）;多西他赛引起的严重不良反应16例次,主要临床表现为过敏样反应（31.25%）、骨髓抑制（18.75%）、白细胞减少（18.75%）、腹泻（6.25%）、过敏性休克（6.25%）、胃肠道出血（6.25%）、背痛（6.25%）、胸闷（6.25%）。停药及对症治疗后均好转或痊愈。结论多西他赛与紫衫醇致严重不良反应临床表现有所不同,与多西他赛相比,紫衫醇过敏性休克所占比例较高,使用时更应加强用药指导和监测。     

酯键链接增韧基团增强姜黄素对肿瘤细胞靶向性作用

目的:以酯键链接增强基团,制备姜黄素前体,观察其对前列腺癌细胞和正常二倍体细胞生长活性影响的差异?方法:叔丁氧羰基(Boc)-苯丙氨酸酯姜黄素单酯(BPC)作用于人前列腺癌DU-145细胞6~24 h后,MTT试验检测细胞生长活性,流式细胞术检测细胞凋亡率?计数法测定1~7天细胞生长曲线?人主动脉平滑肌(people aortic smooth muscle cells,HASMC)细胞作为对照组?结果:10~40 μmol/L的BPC作用于DU-145细胞6~24 h后,DU-145细胞生长抑制率为7.37%~66.87%(P < 0.05),呈浓度﹑时间依赖性;部分细胞出现凋亡形态学改变,FSC-SSC散点图可见24 h DU-145细胞凋亡比率为37.84%~47.12%(P < 0.05)?对照组HASMC细胞凋亡比率为0.94%~4.23%(P < 0.05),较同浓度姜黄素降低?结论:BPC能在体外有效诱导人前列腺癌DU-145细胞凋亡,对正常二倍体细胞的抑制作用较低,为深入研究泌尿系肿瘤靶向性治疗提供了新的途径?     

10-乙酰乙酰紫杉醇标准品的制备工艺研究

目的 研究半合成紫杉醇中工艺杂质10-乙酰乙酰紫杉醇的制备方法。方法 选择性地保护紫杉醇的2''位和7位羟基,然后在碱性条件下脱去10位乙酰基,未保护的10位羟基与2,2,6-三甲基-4H-1,3-二噁英-4-酮进行缩合,最后脱去7位和2''位的保护基,制备得到10-乙酰乙酰紫杉醇。结果 以紫杉醇为原料通过5步反应制备得到10-乙酰乙酰紫杉醇,总收率为27%,纯度高达95%。结论 为10-乙酰乙酰紫杉醇标准品的制备提供了简便易行的方法,有利于紫杉醇原料药的质量控制与评价。