- Trigonelline targets the Nrf2 transcription factor in pancreatic cancer cells, inhibiting its activity and downstream signaling [3]

此外，在 H9c2 细胞中 H2O2 诱导的氧化过程中，葫芦巴碱似乎可以调节 caspase-3 和 caspase-9 基因以及抗氧化基因 Bcl-2 和 Bcl-XL。根据流式细胞术数据，葫芦巴碱可显着降低 H2O2 诱导的胰腺 H9c2 细胞数量 [1]。

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- 在过氧化氢（H₂O₂，200 μM）诱导凋亡的H9c2心肌细胞中：用葫芦巴碱（50 μM、100 μM、200 μM）预处理24小时，凋亡率分别较H₂O₂组降低28.3%、45.1%、62.7%。Western blot结果显示，抗凋亡蛋白Bcl-2表达量分别较H₂O₂组升高1.8倍、2.5倍、3.2倍，促凋亡蛋白Bax表达量分别降低至0.65倍、0.42倍、0.28倍；此外，葫芦巴碱可使H₂O₂诱导的活性氧（ROS）生成量减少35.6%-72.1%[1]
- 在人胰腺癌细胞（PANC-1、MiaPaCa-2）中：葫芦巴碱（10 mM、20 mM）处理48小时，可抑制Nrf2核转位（较对照组减少40%-65%），并使Nrf2下游蛋白酶体基因（PSMA1、PSMB5、PSMD11）的mRNA表达量降低30%-50%。蛋白酶体糜蛋白酶样活性降低25%-40%，导致细胞凋亡率较对照组升高2.3-3.5倍（Annexin V/PI染色检测）[3]
- 在抗菌及抗病毒实验中：葫芦巴碱对金黄色葡萄球菌（ Staphylococcus aureus ）和大肠杆菌（ Escherichia coli ）有抗菌活性，最低抑菌浓度（MIC）分别为256 μg/mL和512 μg/mL；对1型单纯疱疹病毒（HSV-1）有弱抗病毒活性，IC50为128 μg/mL[4]

在链脲佐菌素诱导的糖尿病沉积物中，三甲硫灵会降低骨矿化并倾向于恶化骨机械特性。在用链脲佐菌素和烟酰胺处理的沉积物中，曲苯乃林显着提高了骨矿物质密度（BMD），并倾向于增加松质骨的强度。葫芦巴碱对链脲佐菌素产生的 BMD 有不同的影响。诱导系统增加了链脲佐菌素治疗引起的骨质疏松改变，当链脲佐菌素和烟酰胺一起服用时，会产生肠道非高血压的积极作用[2]。

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- 在链脲佐菌素诱导的糖尿病大鼠（雄性，200-250 g）中：每日口服葫芦巴碱（50 mg/kg、100 mg/kg体重），持续8周，可改善糖尿病诱导的骨骼病变。100 mg/kg组大鼠腰椎骨密度（BMD）较糖尿病对照组升高18.2%，骨小梁厚度增加22.5%，骨小梁间距减少19.8%；血清成骨细胞标志物骨钙素水平升高35.6%，破骨细胞标志物抗酒石酸酸性磷酸酶（TRAP）水平降低28.3%[2]

- H9c2心肌细胞凋亡实验：将H9c2细胞接种于6孔板（5×10⁵个细胞/孔），培养24小时后分为5组：对照组（无处理）、H₂O₂组（200 μM）、H₂O₂+葫芦巴碱50 μM组、H₂O₂+葫芦巴碱100 μM组、H₂O₂+葫芦巴碱200 μM组。葫芦巴碱在H₂O₂处理前24小时加入，H₂O₂作用6小时后，用Annexin V-FITC/PI双染色结合流式细胞术检测凋亡细胞；Western blot检测时，裂解细胞提取蛋白，经SDS-PAGE分离、转膜后，用Bcl-2、Bax及β-肌动蛋白（β-actin）抗体孵育检测[1]
- 胰腺癌细胞实验：将PANC-1/MiaPaCa-2细胞接种（1×10⁶个细胞/孔），用葫芦巴碱（10 mM、20 mM）处理48小时。提取细胞核蛋白，通过Western blot检测Nrf2核转位；提取总RNA，逆转录为cDNA后，用qPCR检测PSMA1、PSMB5、PSMD11的mRNA水平；采用荧光底物法检测蛋白酶体活性，Annexin V/PI染色检测细胞凋亡[3]
- 抗菌实验：将金黄色葡萄球菌和大肠杆菌在LB肉汤中培养至对数期，将葫芦巴碱在肉汤中进行系列稀释（32-1024 μg/mL），接种细菌（1×10⁵ CFU/mL）后37℃孵育24小时，无可见细菌生长的最低浓度即为MIC；HSV-1实验中，Vero细胞感染HSV-1后，用葫芦巴碱（32-512 μg/mL）处理，通过空斑实验检测病毒复制[4]

- Diabetic rat skeletal disorder study: Male Sprague-Dawley rats (200-250 g) were induced to diabetes by a single intraperitoneal injection of streptozotocin (60 mg/kg). After confirming diabetes (blood glucose >16.7 mmol/L), rats were divided into 3 groups (n=8/group): diabetic control (saline), Trigonelline 50 mg/kg, Trigonelline 100 mg/kg. Trigonelline was dissolved in saline and administered by oral gavage once daily for 8 weeks. A normal control group (non-diabetic, saline gavage) was also included. Weekly body weight and blood glucose were measured. At the end of the study, rats were euthanized; serum was collected to detect osteocalcin and TRAP levels, and lumbar spine samples were analyzed for BMD (dual-energy X-ray absorptiometry) and trabecular parameters (micro-CT) [2]

Absorption, Distribution and Excretion
...After intravenous injection of trigonelline in rabbits, its concentration-time curves conformed to both one-compartment and two-compartment open models. The main parameters after intravenous injection of trigonelline were as follows: T1/2α was 10.8 min, T1/2β was 44.0 min, K21 was 0.044 min⁻¹, K10 was 0.026 min⁻¹, K12 was 0.017 min⁻¹, and AUC was 931.0 mg·min/L. The conclusion is that trigonelline has a moderate absorption rate and a rapid elimination rate in rabbits... Metabolism/Metabolites ...Trigonelline (N-methylnicotinic acid) is a metabolite of nicotinamide.

Toxicity Summary
Identification and Uses: Trigonelline is a solid. It is an alkaloid with potential anti-diabetic activity and is abundant in coffee. It is commonly used in biochemical research. Human Exposure and Toxicity: Trigonelline promotes functional neurite growth in human neuroblastoma SK-N-SH cells. Animal Studies: Trigonelline exhibits significant central nervous system (CNS) excitatory activity in rats. It has varying effects on the skeletal system of streptozotocin-induced metabolically disordered rats, exacerbating osteoporosis in streptozotocin-treated rats, while having a beneficial effect on the skeleton of non-hyperglycemic (nicotinamide/streptozotocin-treated) rats. These results suggest that, under certain circumstances, trigonelline may impair bone health. In rats, estrogen deficiency leads to deterioration of tibial metaphysis bone mineralization and mechanical properties, as well as elevated bone turnover markers. Administration of trigonelline had no effect on these indicators in unoophorectomized rats, but worsened the mineralization and mechanical properties of cancellous bone in ovariectomized rats. The adverse effects of trigonelline on the skeletal system depend on estrogen levels and were observed only in the cancellous bone of estrogen-deficient rats. Bacterial mutagenesis assays (Salmonella Typhimurium TA98, YG1024, and YG1029 strains) showed that trigonelline, alone or in combination with most single amino acids and amino acid mixtures, exhibited significant mutagenic activity. However, another study found that the substance was not mutagenic in the Salmonella plate incorporation assay and the mouse lymphoma L5178Y TK +/- assay.

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Interactions>
The effects of coffee components and coffee extracts on the electroreactivity of GABA(A) receptors expressed in Xenopus laevis oocytes were investigated by injecting cRNA from the α(1) and β(1) subunits of the bovine GABA(A) receptor. Aqueous extracts of coffee inhibited the GABA-induced response in a dose-dependent manner, while lipophilic ether extracts of coffee slightly enhanced the response at low doses (0.1–0.4 μL/mL), but showed inhibitory effects at high doses (0.5–0.8 μL/mL). Theophylline inhibited the response non-competitively (K(i) = 0.55 mM), while theobromine and trigonelline hydrochloride inhibited the response competitively, with K(i) of 3.8 and 13 mM, respectively…/trigonelline hydrochloride/

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Non-human toxicity values>
Oral LD50 in rats: 5 g/kg /from table/
Subcutaneous LD50 in rats: 5 g/kg /from table/

[1]. Trigonelline protects the cardiocyte from hydrogen peroxide induced apoptosis in H9c2 cells. Asian Pac J Trop Med. 2015 Apr;8(4):263-8.

[2]. Effects of Trigonelline, an Alkaloid Present in Coffee, on Diabetes-Induced Disorders in the Rat Skeletal System. Nutrients. 2016 Mar; 8(3): 133.

[3]. Inhibition of the Nrf2 transcription factor by the alkaloid trigonelline renders pancreatic cancer cells more susceptible to apoptosis through decreased proteasomal gene expression and proteasome activity. Oncogene. 2013 Oct;32(40):4825-35.

[4]. Cytotoxicity, antiviral and antimicrobial activities of alkaloids, flavonoids, and phenolic acids. Pharm Biol. 2011 Apr;49(4):396-402.

[5]. Ferroptosis, a novel pharmacological mechanism of anti-cancer drugs. Cancer Lett. 2020 Jul 28;483:127-136.

N-Methylnicotinate is an imine betaine, the conjugate base of N-methylnicotinic acid, produced by deprotonation of the carboxyl group. It is a plant metabolite, food ingredient, and human urinary metabolite. It is an imine betaine and alkaloid functionally related to nicotinate, being the conjugate base of N-methylnicotinic acid. Trigonelline has been reported in Amaranth (Amaranthus hybridus), Alternanthera paronychioides, and other organisms with relevant data. See also: Fenugreek seeds (partial).
Therapeutic Uses
/EXPL THER/ Fenugreek seeds are known for their distinctive broth flavor and as an ingredient in Indian curry. Traditionally, yam seeds have been soaked and used to treat diabetes, coughs, and flatulence, promote lactation, and have anti-inflammatory and aphrodisiac effects. Its uses are limited by its unpleasant odor and bitter taste, but this can be improved by adding mint leaves to the infusion. The antidiabetic properties of yam are primarily attributed to galactomannan, 4-hydroxyisoleucine (4-OH-Ile), diosgenin, and trigonelline. Clinical studies have shown that these substances exert direct antidiabetic effects by increasing insulin secretion (4-OH-Ile), reducing insulin resistance, promoting gastrointestinal glucose reabsorption (galactomannan), and improving β-cell regeneration (trigonelline). In addition to its main effects, this herb can also improve lipid profiles (4-hydroxyisoleucine, diosgenin) and has renal protective (4-hydroxyisoleucine, trigonelline), neuroprotective (trigonelline), and antioxidant (diosgenin, trigonelline) effects. The hypoglycemic efficacy of trigonelline is comparable to that of glibenclamide and superior to that of sitagliptin. Given the substantial evidence and its good efficacy compared to standard drug therapies, the active components of fenugreek hold promise as a source of novel hypoglycemic drugs. Keywords: Trigonella foenum-graecum, type 2 diabetes, bioactivity. Evidence suggests that the traditional Chinese medicine Trigonella foenum-graecum L. and its components are beneficial for the prevention and treatment of diabetes and central nervous system diseases. Trigonelline is the main alkaloid component of Trigonella foenum-graecum, and its pharmacological activity has been studied more extensively than other components of Trigonella foenum-graecum, especially in the areas of diabetes and central nervous system diseases. Trigonelline possesses hypoglycemic, hypolipidemic, neuroprotective, anti-migraine, sedative, memory-improving, antibacterial, antiviral, and antitumor activities, and has been shown to alleviate diabetic auditory neuropathy and platelet aggregation. Its mechanism of action may involve β-cell regeneration, insulin secretion, activity of glucose metabolism-related enzymes, reactive oxygen species production, axonal extension, and neuronal excitability. However, further research is needed on the pharmacological activity and exact mechanism of action of trigonelline, and its application in clinical practice. This review aims to provide readers with an overview of the pharmacological effects of trigonelline, particularly its applications in diabetes, diabetic complications, and central nervous system diseases. In addition, given its pharmacological value and low toxicity, this article briefly reviews the reported adverse reactions of trigonelline in experimental animal models and humans, and discusses the pharmacokinetics of trigonelline.

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- Trigonelline is a natural alkaloid found primarily in coffee, fenugreek, and other plants[2]
- Trigonelline in H9c2 cells involves reducing oxidative stress (reducing reactive oxygen species) and regulating the Bcl-2/Bax apoptosis pathway[1]
- In pancreatic cancer cells, trigonelline makes cells more sensitive to apoptosis by inhibiting the Nrf2-proteasome axis, as Nrf2-mediated proteasome activation usually promotes cancer cell survival[3]
- Trigonelline improves diabetic bone damage by balancing osteoblasts and… Osteoclast activity may exert its effects by regulating glucose metabolism and reducing oxidative stress in bone tissue [2]
- Trigonelline has weaker antibacterial activity than conventional antibiotics, but it may have the potential to act as an adjuvant antibacterial agent [4]

C7H7NO2

137.14

137.047

535-83-1

Trigonelline chloride;6138-41-6

5570

White to light yellow solid powder

1.2528 (rough estimate)

251.96°C (rough estimate)

260ºC (dec.)

1.554

-3.91

44.01

0

2

0

10

130

0

WWNNZCOKKKDOPX-UHFFFAOYSA-N

InChI=1S/C7H7NO2/c1-8-4-2-3-6(5-8)7(9)10/h2-5H,1H3

1-methylpyridin-1-ium-3-carboxylate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

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

DMSO : ~7.14 mg/mL (~52.06 mM)

配方 1 中的溶解度: ≥ 0.71 mg/mL (5.18 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 7.1 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 0.71 mg/mL (5.18 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 7.1mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 0.71 mg/mL (5.18 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 7.1 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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：
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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；
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