α-glucosidase

α-葡萄糖苷酶抑制活性[2]
几十年来，研究人员已经证明，桑叶提取物对大鼠和人α-葡萄糖苷酶有很强的抑制作用（Anno et al., 2004, Miyahara et al., 2004, Oku et al., 2006）。α-葡萄糖苷酶位于肠细胞的刷状缘表面膜上，被认为是淀粉和其他碳水化合物消化过程中最重要的酶（Herscovics, 1999）。通过膳食食物和药物改变碳水化合物代谢可能具有治疗价值。1-Deoxynojirimycin (Duvoglustat)/DNJ结合到α-葡萄糖苷酶的活性中心，在小肠中是该酶的有效抑制剂（Junge, Matzke, & Stoltefuss, 1996）。 对于营养保健品的商业开发，应该知道目标化合物及其在产品中的浓度，以达到最佳的治疗效果。在桑树干茶中，我们认为DNJ是关键化合物，因为它能强烈抑制α-葡萄糖苷酶，桑叶中含有高浓度的DNJ（占总亚氨基糖的50%）（Asano et al., 2001）。α-葡萄糖苷酶抑制与纯DNJ (r = 0.96)（图4B）和桑叶DNJ含量（r = 0.84）（图4A）高度相关。在1-Deoxynojirimycin (Duvoglustat)/DNJ浓度下，桑叶提取物对α-葡萄糖苷酶的抑制活性高于DNJ标准：如在5 μg DNJ/ml时，桑叶提取物对α-葡萄糖苷酶的抑制作用为27%，纯DNJ对α-葡萄糖苷酶的抑制作用为23%。这种额外的抑制作用可以解释为桑树提取物中存在其他亚氨基糖（即n -甲基- dnj， 2-O-α-d-半乳糖酰氨基- dnj和fagomine）和其他成分，如异槲皮素，槲皮素和芦丁。

1-Deoxynojirimycin (Duvoglustat)（20 -80 mg/kg；静脉注射；每天一次，持续 4 周）具有抗生理作用[3]。 1-Deoxynojirimycin 通过触发 db/db shark 鲨鱼模式显着增强胰岛素症状。

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1-Deoxynojirimycin (Duvoglustat)/DNJ作为肠道α-葡萄糖苷酶抑制剂广泛用于糖尿病的治疗。然而，关于其对改善胰岛素敏感性的作用的报道很少。本研究的目的是探讨DNJ是否通过改善胰岛素敏感性来降低高血糖。建立了一种制备大量DNJ的经济方法。然后，db/db小鼠静脉注射DNJ(20、40和80 mg·kg（-1）·d(-1)) 4周。通过血糖和生化分析来评价其对高血糖的治疗效果，并探讨骨骼肌的相关分子机制。DNJ显著降低体重、血糖和血清胰岛素水平。DNJ治疗也改善了葡萄糖耐量和胰岛素耐量。此外，虽然骨骼肌中总蛋白激酶B （AKT）、磷脂酰肌醇3激酶（PI3K）、胰岛素受体β (IR-β)、胰岛素受体底物-1 （IRS1）和葡萄糖转运蛋白4 （GLUT4）的表达不受影响，但DNJ处理显著增加了Ser473-AKT、p85-PI3K、Tyr1361-IR-β和Tyr612-IRS1的GLUT4易位和磷酸化。这些结果表明，DNJ通过激活db/db小鼠骨骼肌中胰岛素信号通路PI3K/AKT显著改善胰岛素敏感性。[3]

α-葡萄糖苷酶抑制试验[1]
α-葡萄糖苷酶抑制活性是通过Ma， Hattori， Daneshtalab和Wang（2008）描述的程序的修改来测量的。简单地说，将大鼠肠丙酮粉（1 g）悬浮在100 mM磷酸钾缓冲液（pH 7.0）中，超声振荡20 min。在3000 rpm下离心30 min后，将上清液作为α-葡萄糖苷酶的来源。底物（2 mM 4-硝基苯-α-d-葡萄糖吡喃苷）在100 mM磷酸钾缓冲液（pH 7.0）中移液至96孔板（40 μl/孔）。加入5 μl桑树样品或对照溶液（乙醇与蒸馏水的比例为50:50）混合。加入酶（5 μl）后，37℃孵育20 min，测定405 nm的紫外吸光度。计算桑树样品和标准1-Deoxynojirimycin (Duvoglustat)/DNJ α-葡萄糖苷酶抑制活性的百分比为：（ΔAcontrol-ΔAsample） × 100/ΔAcontrol，其中ΔA为405 nm吸光度。

蛋白质印迹[3]
为了研究1-脱氧野尻霉素/1-Deoxynojirimycin (Duvoglustat)对胰岛素信号通路的影响，如前所述进行了蛋白质印迹分析。简而言之，骨骼肌组织（0.1 g）在裂解缓冲液（50 mM Tris（pH 7.4）、150 mM NaCl、0.1%SDS、0.5%脱氧胆酸钠、1%NP40、10μL磷酸酶抑制剂、1μL蛋白酶抑制剂和5μL 100 mM PMSF）中裂解，在4°C下以16000×g离心15分钟，并通过双辛可宁酸蛋白测定法定量蛋白质浓度。将等量的蛋白质（70μg）装载在10%SDS-PAGE上，并转移到PVDF膜上。膜被阻断后，它们与抗IR-β、对-Tyr1361-IR-β，IRS1、对-Tur612-IRS1、PI3K、对-p85-PI3K、AKT、对-Ser473-AKT、GLUT4、β-actin或Na+K+-ATP酶α1的一抗在4°C下孵育过夜，然后在室温下与HRP偶联的二抗孵育2小时。使用ECL检测试剂盒观察蛋白质条带。以β-actin为对照进行总蛋白表达的正常化。以Na+K+-ATP酶α1为对照进行m-GLUT4表达的正常化。

Animal/Disease Models: db/db mice[3]
Doses: 20, 40, 80 mg/kg
Route of Administration: intravenous (iv) (iv)injection; signal load PI3K/AKT[ 3]. one time/day for four weeks
Experimental Results: Significant reduction in body weight, blood glucose, and serum insulin levels; improved glucose tolerance and insulin tolerance.

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At the end of ten weeks, wild-type C57BLKS mice, which received intravenously normal saline, served as a normal control (N control) (n = 6). The db/db mice were divided into four groups (n = 6): Group I served as a diabetic control and received intravenously normal saline (D control). Group II, III, and IV were treated intravenously with 1-Deoxynojirimycin (Duvoglustat)/DNJ 20, 40, and 80 mg·kg−1·day−1, respectively. An intravenous injection was selected to avoid the function of DNJ as an α-Glycosidase inhibitor inthe gastrointestinal tract. For DNJ doses selection, in our previous study, we screened a large number of Chinese traditional medicines including mulberry leaves by glucose tolerance test of ICR mice. We found the alkaloids (DNJ 40 mg·kg−1) isolated from mulberry leaves could improve the glucose tolerance test of ICR mice (Figure A1). We then tested doses of 10, 20, and 40 mg·kg−1, but both 10 and 20 mg·kg−1 did not have any effect (Figure A2). Therefore, we selected the 1-Deoxynojirimycin (Duvoglustat)/DNJ doses as 20, 40, and 80 mg·kg−1·day−1. All these doses were given for 4 weeks. The blood glucose, body weight and average food intake, water intake, and urine output were measured every week. At the end of the experimental period, the mice were anesthetized with chloral hydrate after withholding food for 12 h, and blood samples were taken to determine the serum insulin levels. Besides, skeletal muscle were removed after the blood was collected, then rinsed with a physiological saline solution, and immediately stored at −80 °C [3].

rat LD50 oral >5 gm/kg BEHAVIORAL: SOMNOLENCE (GENERAL DEPRESSED ACTIVITY); KIDNEY, URETER, AND BLADDER: URINE VOLUME INCREASED; SKIN AND APPENDAGES (SKIN): HAIR: OTHER International Journal of Toxicology., 16(Suppl

[1]. 1-Deoxynojirimycin: Occurrence, Extraction, Chemistry, Oral Pharmacokinetics, Biological Activities and In Silico Target Fishing. Molecules. 2016 Nov 23;21(11). pii: E1600.

[2]. Development of high 1-deoxynojirimycin (DNJ) content mulberry tea and use of response surface methodology to optimize tea-making conditions for highest DNJ extraction. LWT - Food Science and Technology. Volume 45, Issue 2, March 2012, Pages 226-232.

[3]. 1-Deoxynojirimycin Alleviates Insulin Resistance via Activation of Insulin Signaling PI3K/AKT Pathway in Skeletal Muscle of db/db Mice. Molecules. 2015 Dec 4;20(12):21700-14.

Duvoglustat is an optically active form of 2-(hydroxymethyl)piperidine-3,4,5-triol having 2R,3R,4R,5S-configuration. It has a role as an EC 3.2.1.20 (alpha-glucosidase) inhibitor, an anti-HIV agent, an anti-obesity agent, a bacterial metabolite, a hypoglycemic agent, a hepatoprotective agent and a plant metabolite. It is a 2-(hydroxymethyl)piperidine-3,4,5-triol and a piperidine alkaloid.
An alpha-glucosidase inhibitor with antiviral action. Derivatives of deoxynojirimycin may have anti-HIV activity.
1-Deoxynojirimycin has been reported in Parmotrema austrosinense, Parmotrema praesorediosum, and other organisms with data available.
An alpha-glucosidase inhibitor with antiviral action. Derivatives of deoxynojirimycin may have anti-HIV activity.
See also: Fagomine (annotation moved to).

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1-Deoxynojirimycin (DNJ, C₆H13NO₄, 163.17 g/mol), an alkaloid azasugar or iminosugar, is a biologically active natural compound that exists in mulberry leaves and Commelina communis (dayflower) as well as from several bacterial strains such as Bacillus and Streptomyces species. Deoxynojirimycin possesses antihyperglycemic, anti-obesity, and antiviral features. Therefore, the aim of this detailed review article is to summarize the existing knowledge on occurrence, extraction, purification, determination, chemistry, and bioactivities of DNJ, so that researchers may use it to explore future perspectives of research on DNJ. Moreover, possible molecular targets of DNJ will also be investigated using suitable in silico approach.[1]

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Mulberry 1-deoxynojirimycin (DNJ), a potent α-glucosidase inhibitor, suppresses postprandial blood glucose, thereby possibly preventing diabetes mellitus. At present, mulberry dry teas are commercially supplied as functional foods in many countries, but these products may not provide an effective dose (6 mg DNJ/60 kg human wt) due to their low DNJ content (about 100 mg/100 g of dry wt). Therefore, development of tea with higher DNJ content is desirable. To do this, we investigated distribution of DNJ content and α-glucosidase inhibitory activity in 35 Thai mulberry varieties. DNJ content in young leaves varied among mulberry varieties from 30 to 170 mg/100 g of dry leaves. Varieties having highest DNJ content were Kam, Burirum 60 and Burirum 51. Leaf position affected DNJ content: shoots > young leaves > mature leaves. DNJ concentration and α-glucosidase inhibitory activity were highly correlated (r = 0.84), suggesting that α-glucosidase inhibitory activity of mulberry leaves is mainly due to DNJ. Consequently, high DNJ content mulberry tea was produced from shoots of varieties such as Burirum 60, which contains 300 mg/100 g of dry wt. Tea-making conditions were optimized for highest DNJ extraction using response surface methodology. Approximate 95% of total DNJ in high DNJ content dry tea was extracted when temperature was maintained at 98 °C for 400 s; these conditions could be applicable for preparation of commercial products with high DNJ content. One cup (230 ml, a normal serving) of DNJ-enriched mulberry tea contained enough DNJ (6.5 mg) to effectively suppress postprandial blood glucose.[2]

C₆H₁₃NO₄

163.17

163.084

C, 44.17; H, 8.03; N, 8.58; O, 39.22

19130-96-2

1-Deoxynojirimycin hydrochloride;73285-50-4

29435

White to off-white solid powder

1.5±0.1 g/cm3

361.1±42.0 °C at 760 mmHg

195-196°C

197.3±18.5 °C

0.0±1.8 mmHg at 25°C

1.582

-2.1

92.95

5

5

1

11

132

4

O([H])[C@@]1([H])[C@@]([H])([C@]([H])(C([H])([H])N([H])[C@]1([H])C([H])([H])O[H])O[H])O[H]

LXBIFEVIBLOUGU-JGWLITMVSA-N

InChI=1S/C6H13NO4/c8-2-3-5(10)6(11)4(9)1-7-3/h3-11H,1-2H2/t3-,4+,5-,6-/m1/s1

(2R,3R,4R,5S)-2-(Hydroxymethyl)piperidine-3,4,5-triol

BAY-h5595; Duvoglustat; BAY-h-5595; 1-DEOXYNOJIRIMYCIN; 19130-96-2; DUVOGLUSTAT; (2R,3R,4R,5S)-2-(hydroxymethyl)piperidine-3,4,5-triol; Moranolin; D-1-deoxynojirimycin; 1,5-Deoxy-1,5-imino-D-mannitol; (2R,3R,4R,5S)-2-Hydroxymethyl-piperidine-3,4,5-triol; 1-deoxynojirimycin; BAY-h 5595

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ≥ 34 mg/mL (~208.37 mM)

配方 1 中的溶解度: 100 mg/mL (612.86 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。