Antitumor activities of D-glucosamine and its derivatives

The growth inhibitory effects of D-glucosamine hydrochloride （GlcNH2-HCl）, D-glucosamine （GlcNH2） and N-acetyl glucosamine （NAG） on human hepatoma SMMC-7721 cells in vitro were investigated. The results showed that GlcNH2.HCl and GlcNH2 resulted in a concentration-dependent reduction in hepatoma cell growth as measured by MTT （3-（4,5-dimethylthiazol- 2-yl）-2,5-diphenyltetrazolium bromide） assay. This effect was accompanied by a marked increase in the proportion of S cells as analyzed by flow cytometry. In addition, human hepatoma SMMC-7721 cells treated with GlcNH2-HCl resulted in the induction of apoptosis as assayed qualitatively by agarose gel electrophoresis. NAG could not inhibit the proliferation of SMMC-7721 cells. GlcNH2-HCl exhibited antitumor activity against Sarcoma 180 in Kunming mice at dosage of 125-500 mg/kg, dose of 250 mg/kg being the best. GlcNH2-HCl at dose of 250 mg/kg could enhance significantly the thymus index, and spleen index and could promote T lymphocyte proliferation induced by ConA. The antitumor effect of GlcNH2-HCl is probably host-mediated and cytocidal.

Whole-process optimization for industrial production of glucosamine sulfate sodium chloride based on QbD concept

The double salt of glucosamine sulfate sodium chloride(glucosamine-SP) is an important pharmaceuticals ingredient for healing osteoarthritis. However, the study about its industrial production is rarely documented, let alone the optimization over the whole process to produce glucosamine-SP using glucosamine hydrochloride and anhydrous sodium sulfate as synthetic raw materials. In order to improve the production efficiency, this study screened the process parameters based on the concept of quality by design(QbD), optimized 13 operational parameters related to reaction and separation in the process, and finally proposed the mixed dropping process. The reaction conditions for the preparation of glucosamineSP were found as follows: the molar ratio of anhydrous sodium sulfate to glucosamine hydrochloride is 0.42, the mass ratio of water to glucosamine hydrochloride is is 2.0, the reaction temperature is 50 ℃ and the reaction time is 1 h. Through step-by-step scaling up following QbD, the mixed dropping process was successfully applied to achieve a trial production of 200 kg products satisfying national quality standards.In all, the results of this study have high technical value and guiding significance for the industrial mass production of glucosamine-SP.

杏鲍菇废弃菌渣中D-氨基葡萄糖盐酸盐的制备工艺及生物学活性分析

本文以杏鲍菇废弃菌渣为原料,探究了D-氨基葡萄糖盐酸盐(D-glucosamine hydrochloride,GAH)的制备工艺、液相-质谱(HPLC-MS)、红外光谱、理化指标及其对斑马鱼胚胎发育的影响。采用单因素和响应面优化试验,获得盐酸水解制备GAH的最佳条件:盐酸浓度31%,水解时间4 h,水解温度82℃,液固比5 mL/g,此时GAH得率可达23.61%。液相-质谱、红外光谱和理化指标分析显示,GAH纯化样品纯度是标准品的101.9%,质谱和红外光谱图与标准品一致,各项指标均符合甚至优于美国药典43-国家处方集38(USP43-NF38)的质量标准,砷含量仅0.21μg/g。斑马鱼实验结果表明,中高浓度(0.3%~0.5%)的GAH引发的斑马鱼胚胎发育异常比例超过50%,且0.3%的GAH对转化生长因子-β信号通路具有明显的促进作用。本研究结果可为未来杏鲍菇产品产业化生产及应用提供参考。

Effects of Activating Blood Circulation to Resolve Blood Stasis and Glucosamine Capsules Therapy on the Expression of Matrix Metalloproteinase in Post-traumatic Knee Osteoarthritis

[Objectives] To observe the clinical effects of activating blood circulation to resolve blood stasis and glucosamine hydrochloride capsules therapy on post-traumatic knee osteoarthritis treatment,to measure the expression of matrix metalloproteinase in post-traumatic knee osteoarthritis patients before and after treatment,and to explore its relevant molecular mechanisms. [Methods]A total of 60 patients with posttraumatic knee osteoarthritis in early stage in Affiliated Hospital of Shannxi University of Chinese Medicine from January 2015 to December2017 were selected and randomly divided into the control group( n = 30) and the observation group( n = 30). The control group was given oral administration of glucosamine hydrochloride capsules for 6 courses of the treatment. The observation group was using the activating blood circulation to resolve blood stasis therapy for 6 courses of treatment. The recovery of traumatic knee osteoarthritis and the symptom of traditional Chinese medicine( TCM) in 2 groups was compared before and after treatment. The symptoms and the quantitative assessment rating scale of knee osteoarthritis,the pain index of knee and the TCM symptom scores were recorded in both groups. And the synovial fluid and serum of patients were collected to detect the expression of matrix metalloproteinase. [Results]The symptoms and the quantitative assessment rating scale of knee osteoarthritis,the pain index of knee and TCM symptom scores of the 2 groups in the 1 st week,the 12 th week and the 24 th week after the treatment were significantly lower than those before the treatment,and the difference was statistically significant( P < 0. 05). Two therapeutic methods had certain therapeutic effects on the recovery from traumatic knee osteoarthritis,and could significantly improve the TCM syndrome. There were better therapeutic effects on oral decoction of traditional Chinese medicine than oral glycosaminoglycans hydrochloride capsules on treating traumatic knee osteogenesis( P < 0. 05). The expression level of MMP3 was decreased significantly in both groups( P <0. 001),while the expression level of MMP13 had no significant change in both groups( P >0. 05). The expression level of MMP1 was decreased significantly in the observation group( P < 0. 001),but there was no significant change in the control group after treatment( P >0. 05). [Conclusions]Two treatment methods have been effective in this clinical study. We found that the expression level of MMP3 was decreased significantly after treatment in 2 therapies,which can be clinically applied.

滑膜成纤维细胞中盐酸氨基葡萄糖含量的高效液相色谱法测定

目的建立一种快速、准确测定滑膜成纤维细胞中盐酸氨基葡萄糖含量的高效液相色谱法。方法采用乙腈萃取-超声-离心的方法提取滑膜成纤维细胞中盐酸氨基葡萄糖;SB-C18色谱柱分离,乙腈(10 mmol/L)∶戊烷磺酸钠(10∶90)做流动相,柱温常温,流速0.8 mL/min,检测波长192 nm。结果该方法在0.5~2.5 mg/mL浓度范围内具有良好的线性关系(r>0.999);滑膜成纤维细胞中盐酸氨基葡萄糖检出限为0.11 mg/mL;定量限为0.23 mg/mL;回收率为88.7%~95.4%。结论该方法简便、快速、灵敏,是一种检测滑膜成纤维细胞中盐酸氨基葡萄糖含量的优良方法。

氨糖美辛肠溶片中盐酸氨基葡萄糖有关物质研究

目的 建立测定氨糖美辛肠溶片中盐酸氨基葡萄糖有关物质的高效液相色谱法,并分析其影响因素。方法 色谱柱为纳谱ChromCore AQ C18柱(250 mm×4.6 mm,3μm),流动相为水-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为280 nm,柱温为30℃,进样量为20μL。采用加速稳定性试验和辅料相容性试验,结合生产工艺分析有关物质的影响因素。结果 已知杂质果糖嗪、2,5-脱氧果糖嗪、吡嗪、5-羟甲基糠醛、糠醛、2-甲基吡嗪、吡咯-2-甲醛、5-甲基糠醛质量浓度分别在0.20~3.30μg/mL、0.20~3.00μg/mL、0.25~7.54μg/mL、0.18~7.04μg/mL、0.25~10.10μg/mL、0.25~10.18μg/mL、0.25~10.17μg/mL、0.26~10.50μg/mL范围内与峰面积线性关系良好(R2≥0.999);检测限为0.3~5.0μg/mL,定量限为0.7~20.1μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%;平均加样回收率为98.24%~101.20%,RSD为0.33%~2.19%(n=9)。影响制剂有关物质产生的因素为工艺、温度和时间,影响有关物质含量增长速率的因素除工艺外,还有辅料(主要为羟甲基淀粉钠)和水分。结论 该方法简便、准确、专属性好,可用于氨糖美辛肠溶制剂中盐酸氨基葡萄糖有关物质的检测。建议优化工艺中干燥温度及时间,考察添加辅料的合理性,并控制片剂中的水分,进一步提高制剂质量的均一性和稳定性。

盐酸氨基葡萄糖片联合中药制剂制香片预防及治疗轻中度膝骨关节炎的效果观察

目的观察盐酸氨基葡萄糖片联合中药制剂制香片预防及治疗轻中度膝骨关节炎(KOA)的效果。方法选取2021年5月至2023年12月四川省骨科医院收治的110例轻中度KOA患者。完全随机分为常规组和联合组,各55例。其中常规组给予盐酸氨基葡萄糖片治疗,联合组在常规组的基础上加用中药制剂制香片治疗。比较2组患者膝关节疼痛视觉模拟量表(VAS)评分、西安大略和麦克马斯特大学骨关节炎指数(WOMAC)评分、膝关节功能、日常生活活动能力、临床疗效及不良反应。结果治疗后2组VAS评分均低于治疗前且联合组明显低于常规组[(2.4±0.6)分比(3.2±1.3)分],差异有统计学意义(t=3.491,P=0.003)。治疗后联合组WOMAC评分低于常规组,Lysholm膝关节功能评分中活动范围评分大于常规组,差异均有统计学意义(均P<0.05)。治疗6个月时联合组日常生活活动能力量表评分高于常规组[(95±5)分比(77±5)分],差异有统计学意义(t=19.986,P<0.001)。联合组总有效率高于常规组[90.9%(50/55)比76.4%(42/55)],差异有统计学意义(χ^(2)=4.251,P=0.039)。2组均无严重不良反应发生。结论盐酸氨基葡萄糖片联合中药制剂制香片预防及治疗轻中度KOA可有效促进患者疼痛缓解,改善日常生活活动能力和膝关节功能,减少膝关节活动功能受限,安全有效。

中西医结合治疗腰椎小关节骨关节炎临床观察

目的 外搽舒活酒外贴伤科止痛膏联合口服盐酸氨基葡萄糖配合运动治疗腰椎小关节骨关节炎的临床疗效观察。方法 选择145例腰椎小关节骨关节炎患者,随机分为试验组73例、对照组72例。对照组予红外线照射、电针、外贴膏药治疗,观察组在对照组的基础上增加外搽舒活酒外贴伤科止痛膏联合口服盐酸氨基葡萄糖片配合运动治疗;对比分析治疗前、治疗后6周、治疗后12周、治疗周期结束后3个月的CDTY评分及临床疗效。结果 治疗周期结束后3个月,试验组的CDTY评分、痊愈率、显效率均优于对照组(P<0.05)。结论 外搽舒活酒外贴伤科止痛膏联合口服盐酸氨基葡萄糖配合运动治疗腰椎小关节骨关节炎能明显改善症状,且远期疗效明显,值得临床推广。

D-氨基葡萄糖盐酸盐的制备及抗氧化特性的研究

从盘锦河蟹壳制备甲壳素,后者用盐酸水解,再纯化得到D-氨基葡萄糖盐酸盐,同时研究了其对羟自由基(·OH)清除作用和对超氧(O_2^-·)自由基的抑制效果。

D-氨基葡萄糖及其衍生物抗肿瘤活性的初步研究

目的 观察氨基葡萄糖盐酸盐（GlcNH2·HCl）、氨基葡萄糖（GlcNH2）和N-乙酰氨基葡萄糖（NAG）体外抑制人肝癌细胞（SMMC-7721）生长的作用，对小鼠S18。肉瘤的肿瘤抑制作用以及对免疫系统的影响并探讨其作用机制。方法 采用MTT法检测不同浓度的单糖对SMMC-7721细胞生长的影响，倒置显微镜观察细胞形态，琼脂糖凝胶电泳进行DNA断裂分析，流式细胞仪检测对细胞周期的影响。采取灌胃给药的方式，观察GlcNH2·HCl对S18。小鼠瘤重、胸腺重和脾重以及荷瘤小鼠淋巴细胞转化率的影响。结果 GlcNH2·HCl和GlcNH2能明显抑制肝癌细胞SMMC-7721的生长，且存在剂量依赖性，在500μg·mL^-1浓度下，Glc-NH2·HCl和GlcNH2的抑制率为50．24％和52．19％，在1000μg·mL^-1浓度下的抑制率分另0为82．21％和83．2％。GlcNH2·HCl对SMMC-7721细胞存在周期特异性，可以使细胞发生S期阻滞。NAG对肝癌细胞的生长没有明显抑制作用。GlcNH2·HCl在125～500mg·kg^-1的剂量范围内，对小鼠S180肉瘤均有一定的抑制作用，平均抑瘤率在27．84％～34．02％之间，其中250mg·kg^-1剂量的抑制作用最强，在该剂量范围下，GlcNH2·HCl对荷瘤小鼠具有明显增加胸腺指数和脾指数的作用，同时促进荷瘤小鼠脾脏T淋巴细胞转化，无明显毒副作用。结论 GlcNH2·HCl体外呈剂量依赖性抑制人肝癌细胞SMMC-7721的生长，体内可能是通过直接杀伤肿瘤细胞的细胞毒作用和提高S180肉瘤小鼠细胞免疫水平来发挥抑瘤作用。

D-氨基葡萄糖盐酸盐的制备优化

目的优化从甲壳素酸水解制备氨基葡萄糖盐酸盐的制备工艺。方法采用正交试验L9(34),优化水解工艺条件。结果以30%HCl,温度95℃,时间3h效果最佳。纯度>99.0%。结论该优化条件能得到较高产率和纯度的盐酸氨基葡萄糖。

利用斑马鱼肝癌模型探究氨基葡萄糖盐酸盐对肝癌的影响

目的:探究双孢蘑菇来源的氨基葡萄糖盐酸盐(Glucosamine Hydrochloride,GAH)对肝癌的影响。方法:利用斑马鱼肝癌及血管双转基因模型Tg(flk:mCherry;kras^(G12V)),在60 mg/mL盐酸多西环素溶液的诱导下,通过激光共聚焦显微镜追踪观察第5 d到第10 d肝癌的发生发展;在Dox诱导的同时加入不同浓度GAH恢复,在激光共聚焦显微镜下比较分析正常组(Ctr)、模型组(Doxorubicin,Dox)、Dox+0.1%GAH组、Dox+0.3%GAH组中GAH对肝癌及肝癌血管生长的影响;通过qPCR和ELISA实验分析GAH对肝癌及肝癌血管相关调控因子mRNA和蛋白水平表达的影响(prmt5、tiam1、kat5、vegfaa、vegfr2、tgf-β1);最后通过TIMER2在线生物信息学分析验证tgf-β1与prmt5、tiam1、kat5与vegfaa、vegfr2的相关性。结果:Dox持续诱导会引起肝癌的发生发展;与Dox模型组相比,GAH处理极显著抑制肝癌的生长与侵袭(P<0.001),并能极显著改善肝癌血管紊乱(P<0.001);qPCR和ELISA实验结果表明GAH能够显著抑制肝癌及血管相关基因的mRNA和蛋白水平(P<0.05);生物信息学分析结果提示tgf-β1与肝癌及血管相关基因的表达具有明显相关性。结论:GAH可能通过调控TGF-β信号通路影响肝癌相关基因的表达抑制肝癌血管的生成,从而抑制肿瘤的生长与侵袭。

1,3,4,6-四-O-乙酰基-β-D-氨基葡萄糖的合成

分别用对甲氧基苯甲醛和苯甲醛保护D 氨基葡萄糖的氨基 ,再将羟基用乙酰基保护 ,以氯化氢脱去氨基上的保护基团 ,最后脱去氯化氢得到 1,3,4 ,6 四 O 乙酰基 β D 氨基葡萄糖。两种方法的总产率分别为 5 9%和6 1%。道关键中间体 3b未见报 。

2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖的合成

以D 氨基葡萄糖盐酸盐和丁二酸酐为原料 ,采用简便的方法合成 2 (3 羧基 1 丙酰氨基 ) 2 脱氧 D 葡萄糖 ,产率大于 70 % ,结构经元素分析。

HPLC示差折光分析法测定D-氨基葡萄糖盐酸盐中的N-乙酰-D-氨基葡萄糖

利用高效液相示差折光分析法测定D-氨基葡萄糖盐酸盐中的N-乙酰-D-氨基葡萄糖.方法采用Alltech C18柱（250×4.6 mm，5μm），流动相为乙腈-醋酸盐缓冲液（3∶97），流速1 mL/min，柱温40℃，进样量20μL.N-乙酰-D-氨基葡萄糖在4.18-1 079.33μg/mL范围内呈显著的线性关系，相关系数为0.999 3.方法的加标回收率98.78%，检出限4.18μg/mL.

滴定分析法和分光光度法测定保健食品中D-氨基葡萄糖盐酸盐含量的研究

〔目的〕建立保健食品中D-氨基葡萄糖盐酸盐测定方法。〔方法〕采用滴定分析方法和分光光度法,定量测定保健食品中的D-氨基葡萄糖盐酸盐。〔结果〕滴定分析法精密度（n=6）为94.0％±0.4％,相对标准偏差（RSD）为0.4％回收率为97.1％～99.7％。分光光度法精密度（n=6）为93.9％±1.0％,相对标准偏差（RSD）1.1％,回收率为97.4％～103.0％。对两种实验方法比对实验的结果进行t检验,t=0.243<t（0.05,18）=2.101,P>0.05,两种分析方法所得实验的结果无明显差异。〔结论〕两种方法操作简便,精密度、准确度均较高,适合于保健食品中D-氨基葡萄糖盐酸盐的测定,滴定分析法更适合D-氨基葡萄糖盐酸盐含量较高的保健食品及原料纯度进行测定。

1,3,4,6-四-O-乙酰基-β-D-氨基葡萄糖合成方法的改进

以氨基葡萄糖盐酸盐为原料,在乙酸酐和浓硫酸的作用下以71%的收率合成了1,3,4,6-四-O-乙酰基-α-D-氨基葡萄糖硫酸盐(2),然后再用无水乙酸钠脱除其中的硫酸,以90%的收率得到1,3,4,6-四-O-乙酰基-β-D-氨基葡萄糖(3).通过1H NMR验证产品的结构,并对2和3的构型进行了分析,对脱除硫酸反应中溶液pH值的变化做了讨论.

甲壳素超微粉碎辅助制备D-氨基葡萄糖盐酸盐

研究了甲壳素超微粉碎辅助制备D-氨基葡萄糖盐酸盐(GAH)的工艺。对HCl浓度、反应时间、反应温度和液料比进行单因素考察,根据考察结果进行Box-Benhnken中心组合设计,利用响应面分析法优化工艺参数,然后进行验证实验,并将该工艺条件与传统GAH制备工艺比较。结果表明,甲壳素超微粉碎辅助制备GAH的较佳工艺为:HCl浓度8.8mol/L、液料比4.2∶1(mL∶g)、反应时间3.6h、反应温度83.0℃。此条件下GAH得率为74.82%,与理论值基本相符。与传统制备工艺相比,新工艺反应条件更加温和,同时GAH得率提高了14.77%。

N-乙酰氨基葡萄糖及D-氨基葡萄糖盐酸盐研究进展

随着对甲壳素及其衍生物的不断关注,现已经形成甲壳素化学这一学科,本文着重围绕甲壳素化学体系结构中的N-乙酰D-氨基葡萄糖和D-氨基葡萄糖盐酸盐两种物质在生物领域中的应用和研究作简要概述。

相转移催化法合成1-O-(2,4-二硝基苯)-2-乙酰氨基-β-D-葡萄糖苷

This paper reports that in the presence of cetyltrimethyl ammonium bromide as phase transfer catalyst,D-glucosamine hydrochloride reacts with 2,4-dinitrophenol to synthesize 1-O-(2’,4’-dinitrophenyl)-2-acetamido-β-D-glucoside.Chemical structure of the new compound was confirmed by elemental analysis,IR and UV.