A systematic review of pharmacological activities, toxicological mechanisms and pharmacokinetic studies on Aconitum alkaloids

The tubers and roots of Aconitum(Ranunculaceae)are widely used as heart medicine or analgesic agents for the treatment of coronary heart disease,chronic heart failure,rheumatoid arthritis and neuropathic pain since ancient times.As a type of natural products mainly extracted from Aconitum plants,Aconitum alkaloids have complex chemical structures and exert remarkable biological activity,which are mainly responsible for significant effects of Aconitum plants.The present review is to summarize the progress of the pharmacological,toxicological,and pharmacokinetic studies of Aconitum alkaloids,so as to provide evidence for better clinical application.Research data concerning pharmacological,toxicological and pharmacokinetic studies of Aconitum alkaloids were collected from different scientific databases(PubMed,CNKI,Google Scholar,Baidu Scholar,and Web of Science)using the phrase Aconitum alkaloids,as well as generic synonyms.Aconitum alkaloids are both bioactive compounds and toxic ingredients in Aconitum plants.They produce a wide range of pharmacological activities,including protecting the cardiovascular system,nervous system,and immune system and anti-cancer effects.Notably,Aconitum alkaloids also exert strong cardiac toxicity,neurotoxicity and liver toxicity,which are supported by clinical studies.Finally,pharmacokinetic studies indicated that cytochrome P450 proteins(CYPs)and efflux transporters(ETs)are closely related to the low bioavailability of Aconitum alkaloids and play an important role in their metabolism and detoxification in vivo.

Microcalorimetry studies of the antimicrobial actions of Aconitum alkaloids

The metabolic activity of organisms can be measured by recording the heat output using microcalorimetry. In this paper, the total alkaloids in the traditional Chinese medicine Radix Aconiti Lateralis were extracted and applied to Eschenchia coil and Staphylococcus aureus. The effect of alkaloids on bacteda growth was studied by microcalorimetry. The power-time curves were plotted with a thermal activity monitor （TAM） air isothermal microcalorimeter and pa- rameters such as growth rate constant （p）, peak-time （Trn）, inhibitory ratio （I）, and enhancement ratio （E） were cal- culated. The relationships between the concentration of Aconitum alkaloids and p of E. coil or S. aureus were discussed. The results showed that Aconitum alkaloids had little effect on E. coil and had a potentially inhibitory effect on the growth of S. aureus.

Effect of processing on the alkaloids in Aconitum tubers by HPLC-TOF/MS

According to the Chinese Pharmacopoeia 2015, only processed Aconitum tubers can be clinically applied, and the effect of processing is unclear. This research aimed to explore the effect of processing on cardiac efficacy of alkaloids in Aconitum tubers. First, the chemical ingredients in unprocessed and processed Aconitum tubers were identified and compared by using high performance liquid chromatography time-of-flight mass spectrometry(HPLC-TOF/MS) and multivariate pattern recognition methods. Then the representative alkaloids in Aconitum tubers, aconitine, benzoylaconine, and aconine, which belong to diester-diterpenoid alkaloids,monoester-diterpenoid alkaloids, and amine-diterpenoid alkaloids, respectively, were selected for further validation of attenuated mechanism. Subsequent pharmacological experiments with aconitine, benzoylaconine,and aconine in SD rats were used to validate the effect of processing on cardiac functions. After processing the Aconitum tubers, it was found that the contents of diester-diterpenoid alkaloids were reduced, and those of monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids were increased, suggesting that diesterditerpenoid alkaloids were transformed into monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids.Through further decocting the aconitine in boiling water, it was confirmed that the three alkaloids could be progressively transformed. Pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats showed that aconitine at a dose of 0.01 mg/kg and aconine at a dose of 10 mg/kg enhanced the cardiac function, while benzoylaconine at a dose of 2 mg/kg weakened the cardiac function. The effect of processing is attributed to the transformation of the most toxic diester-diterpenoid alkaloids into less toxic monoesterditerpenoid alkaloids and amine-diterpenoid alkaloids.

Two New Norditerpenoid Alkaloids from Aconitum spicatum Stapf

Two new norditerpenoid alkaloids, spicatine A (1) and spicatine B (2) were isolated from the root of Aconitum spicatum. The new compounds were deduced on the basis of their spectral data (IR, HREIMS, EIMS, 1D, 2D-NMR). This is the first whole report on the isolation of diterpenoid alkaloids from the A.spicatum Stapf.

Two New Norditerpenoid Alkaloids of Aconitum nagarum

Two new norditerpenoid alkaloids were isolated from the roots of Aconitum nagarum.Their structures were elucidated as 13-hydroxyfranchetine 1 and 10-dehydroxyflavaconitine 2,respectively, by 1D and 2D NMR experiments.

Two new C18-norditerpenoid alkaloids from Aconitum delavayi

Further phytochemical investigation of the unique C 18-norditerpenoid alkaloids from the roots ofAconitum delavayi Franch led to the isolation of two new norditerpenoid alkaloids, delavaconitine F 1 and delavaconitine G 2. Their structures were determined from spectroscopic evidence.

Analysis of Norditerpenoid Alkaloids Extracted from Aconitum sinomantanum Nakai by Electrospray Ionization Tandem Mass Spectrometry

Electrospray ionization mass spectrometry（ESI-MS） was applied simultaneously in determining norditerpenoid alkaloids from the roots of Aconitum sinomantanum Nakai （RAS） based on molecular mass information. The tandem mass spectra（ ESI-MS^n） provided the alkaloidal structural information, through which the existence of these alkaloids was further confirmed. Accordingly, six known norditerpenoid alkaloids were simultaneously determined on the basis of their ESI-MS^n spectra. Furthermore, based on the diagnostic fragmentation pathways of alkaloidal MS^n, a rapid method for direct detection and characterization of alkaloids from an ethanolic extract of RAS was described.

Diterpenoid alkaloids from a Tibetan medicinal plant Aconitum richardsonianum var. pseudosessiliflorum and their cytotoxic activity

The chemical constituents from Aconitum richardsonianum var.pseudosessiliflorum were investigated.The roots of this plant were extracted three times with 90% EtOH at the room temperature.The ethanol extracts were combined and concentrated under reduced pressure to yield residue,which was suspended in water and successively partitioned with chloroform.The chloroform extraction was isolated and purified by silica gel and Sephadex LH-20 column chromatography.Six compounds were isolated and elucidated as delelatine（1）,isodelpheline（2）,3-acetylaconitine（3）,isoatisine（4）,nordhagenine A（5）and yunaconitine（6）.Compounds 1-5 were obtained from Aconitum Brunneum for the first time.Compound（1）showed significant cytotoxic activities（IC50=4.36 μM）against the human tumor cell line P388.

Matrix Solid-phase Dispersion Extraction of Alkaloids from the Roots of Aconitum kusnezoffii Reichb

Matrix solid-phase dispersion（MSPD） was developed for the extraction of four alkaloids, including aconitine, mesaconitine, hypaconitine and deoxyaconitine, from the roots ofAconitum kusnezoffii Reichb. The determination of the analyte was carried out by high performance liquid chromatography with UV detection. The alkaline alumina was used as sorbent. The mixture of acetonitrile and water was used as elution solvent. Several extraction parameters, such as type of sorbent, the ratio of sample to solid support material, type of the elution solvent and the volume of the elution solvent were tested. Mean recoveries ranged from 93.16% to 102.73%, with relative standard deviations from 0.27% to 4.17%. With the extraction efficiency and time expenditure taken into account, MSPD extraction should be a comparatively good method.

Two new C_(19)-diterpenoid alkaloids from roots Aconitum hemsleyanium var. atropurpureum

A new franchetine-type C19-diterpenoid alkaloid 3-hydroxyfranchetine 1 and a new aconitine-type C19-diterpenoid alkaloid atropurpursine 2 have been isolated from the roots of Aconitum hemsleyanium var. atropurpureum. The structures of these new alkaloids were established on the basis of spectral data.

Investigation of Aconitine-type Alkaloids from Processed Tuber of Aconitum carmiechaeli by HPLC-ESI-MS/MS^n

Introduction Aconitine-type alkaloids isolated from the roots of Aconitum carmiechaeli show a potential toxicity and a broad spectrum of bioactivity[1-4].On the basis of the C8-substituent of C19-diterpenoid skeleton,aconitine-type alkaloids can be divided into diester-diterpenoid alkaloids（DDAs）,monoester-diterpenoid alkaloids（MDAs）,and lipo-alkaloids（Fig.1）.

乌头属中药的毒性及毒代动力学研究进展

包括草乌、川乌、附子等在内的乌头属中药因其药理作用独特、药理活性显著,长期以来被用作治疗疾病的药物,或者作为一些新药开发设计的出发点,因此,乌头属中药应用广泛,具有一定的研究价值。尽管乌头属中药的药效显著,有难以替代的疗效,但其具有的毒性作用依旧不容忽视,对其的临床使用使得医、患担心,因此人们长期以来都在尽力研究乌头安全使用的方法和手段,致力于挖掘运用中药乌头独特的药理活性的同时,也在深入研究预防乌头属植物中毒的手段。毒代动力学的研究是评估药物药效、毒性作用、药物安全性评估及中毒推断的一种重要手段,用以解释药物暴露毒性实验结果和预测人用药物治疗不良反应的方法之一。因此,综述了中药乌头的毒性和毒代动力学研究方法,以期为乌头属中药的增效减毒的研究,如降低心脏毒性,保留其抗风湿性关节炎等药理活性,安全使用和中毒法医相关推断的研究提供参考,并对乌头属植物的使用前景进行展望。

乌头生物碱及其代谢物在家兔体内的死后弥散规律

目的探究乌头生物碱及其代谢物在中毒家兔体内的死后弥散规律,为乌头生物碱中毒相关死亡案件生前服毒或死后染毒的判别提供参考依据。方法将24只家兔经气管夹闭处死1 h后,以生川乌水煎液灌胃(以生川乌水煎液中乌头碱的LD50计算)。灌胃后将家兔以仰卧位保存于25℃气候箱中,分别于0 h、4 h、8 h、12 h、24 h、48 h、72 h、96 h随机各取3只进行解剖,取心血、外周血、尿液、心脏、肝、脾、肺和肾组织,采用高效液相色谱-串联质谱法对乌头生物碱及其代谢物在各生物检材中的含量进行检测。结果灌胃后4 h,乌头生物碱及其代谢物在心血、外周血和主要器官组织中均可检出,且含量随保存时间的延长发生动态变化。其中,脾、肝和肺中乌头生物碱及其代谢物含量均较高,尤其距离胃较近的脾中乌头生物碱及其代谢物含量最高。灌胃后48 h,家兔脾中代谢物苯甲酰新乌头原碱的平均质量分数最高;肾中乌头生物碱及其代谢物含量均偏低;尿液中未检出乌头生物碱及其代谢物。结论乌头生物碱及其代谢物在中毒家兔体内存在死后弥散现象,且从含量高的器官(胃)扩散到其他主要器官组织及心血中,其主要机制是顺浓度梯度的弥散,而尿液不受死后弥散的影响,可考虑将其作为乌头生物碱生前服毒和死后染毒的鉴别依据。

雪上一枝蒿生物碱可溶性微针制备

目的 制备雪上一枝蒿生物碱可溶性微针。方法 离心注模法制备可溶性微针。在单因素试验基础上,以硫酸软骨素用量、PVP K120用量、40%乙醇用量为影响因素,刺孔率为评价指标,Box-Behnken响应面法优化处方,考察形态、穿刺性能、含药量、体外透皮性能。结果 最优处方为硫酸软骨素用量123 mg, PVP K120用量298 mg, 40%乙醇用量2.4 mL,刺孔率为98.3%。可溶性微针为黄色正方形贴片,针体呈圆锥形,可刺穿铝箔和大鼠皮肤,含药量为(0.94±0.025)mg。可溶性微针组24 h内累积渗透率为91.4%,明显高于凝胶膏剂组,渗透符合Higuchi方程。结论 雪上一枝蒿可溶性微针机械强度较好,可实现药物的有效经皮递送。

乌头不同叶片中化学成分的变化特征及打顶对根的影响

目的基于乌头传统栽培中打顶的生产工艺,对乌头不同位置叶片所含的化学成分进行系统分析,并对打顶前后根进行质量及成分比较,以明确乌头叶片中化学成分的分布,为打顶工艺提供科学依据。方法采用UPLC-Q-TOF-MS技术,以乙腈-0.1%甲酸水为流动相,柱温45℃,流速0.4 ml/min,对乌头顶叶、不同部位的叶片(前6片叶)、打顶前后根部样品中二萜生物碱类成分进行检测,并通过多元统计分析打顶工艺对乌头的影响。结果顶叶中分布着含量与数量极其丰富的二萜生物碱类成分,在顶叶检测出60种化学成分,其中25个为顶叶中的特异性成分,特异性成分类型为单酯型、双酯型及三酯型二萜生物碱。与顶叶比较,相对成熟的叶片(第1~6片)二萜生物碱类成分的个数明显降低,且不同类型化合物降低的程度依次为三酯型≈双酯型>单脂型>醇胺型,即双酯型及三酯型二萜生物碱的含量在成熟叶片中迅速降低。对乌头进行打顶处理后,根的产量增长10%/个,多元统计分析表明打顶前后地下根中存在Songorine、7-deoxylycoctonine、Fuziline、chasmanine、14-acetyltalatizamine、10-OH-mesaconitine、mesaconitine等主要差异成分。结论乌头顶叶中存在大量且特异的二萜生物碱类成分,打顶工艺对不同部位二萜生物碱的分布有影响且对乌头的增产有重要作用。

不同盐浸处理对附子盐制工艺的影响

为了规范附子盐浸工艺,通过研究鲜附子经4种胆巴替代盐的不同浓度溶液浸泡处理后的生物碱含量变化,对比获得盐浸加工的最佳的盐种类、盐溶液浓度及浸泡时间,筛选出最佳仿“浸胆”工艺,为附子盐浸工艺的质量控制提供科学依据。鲜附子经4种不同种类盐(CaCl_(2)、MgCl_(2)、NaCl、CaCl_(2)∶MgCl_(2)(1∶1))的不同质量分数盐溶液浸泡处理7~80 d,定期采样,采用HPLC法测定6种酯型生物碱含量,分析不同盐浸处理对其产生的主要影响及变化规律,以生物碱含量及毒性成分指数综合筛选最优工艺。通过比较酯型生物碱损失量及防腐效果,加工所用的各种盐的最佳质量分数分别为35%CaCl_(2)、30%MgCl_(2)、25%NaCl、35%CaCl_(2)∶MgCl_(2)(1∶1),随浸泡时间延长,附子双酯型生物碱总含量呈持续下降趋势。与鲜附子相比,不同种类、不同浓度的盐溶液浸泡处理显著影响附子中酯型生物碱类成分的组成及含量。鲜附子经过以上浓度的各种盐浸处理,防腐、减毒效果均可达到最佳。综合分析附子饮片实际生产中盐浸处理的时长、双酯型生物碱的减损程度、毒性成分指数和生产用盐辅料的成本,研究得出35%CaCl_(2)为附子盐浸工艺的最优盐溶液,并建议盐浸处理时间控制在7~21 d之内。

基于物质基础的雪上一枝蒿饮片及其水提物体内安全性比较研究

目的建立HPLC同时检测雪上一枝蒿饮片6个乌头类生物碱含量的方法,比较饮片及其水提干膏粉的急性毒性反应特征和严重程度差异,为雪上一枝蒿饮片质量标准提高及合理使用提供依据。方法采用ThermoAcclaim 120 C18色谱柱,以乙腈-四氢呋喃(5∶3,V/V)和0.1 mol/L醋酸铵溶液为流动相,梯度洗脱,柱温30℃,流速1.0 mL/min,测定雪上一枝蒿饮片及其水提干膏粉6个生物碱含量。将100只健康ICR小鼠随机分为溶媒对照组(0.5%羧甲基纤维素钠溶液),雪上一枝蒿饮片6.50、7.65g/kg组及水提干膏粉10.33、12.15 g/kg组,给予相应药物后观察小鼠14 d内的中毒表观特征及死亡情况。结果雪上一枝蒿炮制饮片及其水提干膏粉中苯甲酰乌头原碱的含量为0.10%和0.42%,其余5个生物碱均未检出。毒性试验结果表明,雪上一枝蒿饮片小鼠最大耐受剂量为6.50g/kg(相当于临床人拟用剂量的7831倍),其水提干膏粉小鼠最小致死剂量为30.99g/kg(相当于临床人拟用剂量的193688倍)。结论本研究建立的HPLC同时测定雪上一枝蒿饮片及其水提干膏粉6个乌头类生物碱含量的方法线性关系良好且准确性高,雪上一枝蒿饮片水提干膏粉乌头类生物碱含量更高且安全性更好。

雪上一枝蒿总碱及其主要二萜生物碱的镇痛活性研究

分别采用醋酸扭体法和热板法研究雪上一枝蒿总碱及5个主要二萜生物碱成分乌头碱、3-去氧乌头碱、尼奥灵、12-表-15-乙酰欧乌头碱、12-表-欧乌头碱外周和中枢的镇痛活性.与空白对照组比较,雪上一枝蒿总碱高、中剂量组,乌头碱高、中、低剂量组,尼奥灵高、中剂量组,12-表-15-乙酰欧乌头碱的高剂量组和12-表-欧乌头碱的中剂量组对冰醋酸所致的小鼠扭体均具有抑制作用,差异具有统计学意义(P<0.05或P<0.01);与本组基础痛阈值比较,雪上一枝蒿总碱高、中、低剂量组,乌头碱高、中、低剂量组,3-去氧乌头碱高剂量组,尼奥灵、12-表-15-乙酰欧乌头碱以及12-表-欧乌头碱的高、中剂量组在给药15 min后均能显著延长热板所致小鼠疼痛的痛阈值(P<0.05或P<0.01).由此可得,雪上一枝蒿总碱及5个主要二萜生物碱成分均有明显的镇痛作用.

雪上一枝蒿生物碱水凝胶微针的制备及体外透皮研究

采用化学交联法和离心浇注法制备雪上一枝蒿生物碱水凝胶微针(HMN-AAB)并考察其理化性质及体外透皮性能。单因素考察水凝胶微针的基质用量、干燥条件。通过正交实验优化微针基质用量,最终确定微针的制备工艺参数。采用高效液相色谱(HPLC)法测定其载药量,利用扫描电子显微镜观察其形貌,通过苏木精-伊红(HE)染色法分析其穿刺性能,并结合Franz扩散池考察其体外透皮性能。HMN-AAB的最佳制备参数为:聚乙烯马来酸酐共聚物2 g,聚乙二醇10000(PEG10000)0.75 g,无水碳酸钠0.3 g。制备的HMN-AAB的载药质量分数为4%。采用离心浇注法制备微针,通过程序升温进行干燥。HMN-AAB为正方形贴片,表面光滑,阵列完整,微针针尖呈圆锥形,含药量为(4.50±0.08)mg/片(以雪上一枝蒿甲素计)。体外透皮结果显示,48 h后HMN-AAB中药物的累积渗透率可达93%,明显高于普通凝胶膏剂组,释放曲线符合Higuchi方程。制备的HMN-AAB机械强度好、载药量大,实现了AAB的透皮递送。