Bupivacaine HCl (HSDB 7790) primarily targets voltage-gated sodium channels [1]
It also exerts antitumor effects on gastric cancer cells via targets independent of sodium channel blockade [2]

在与中枢敏化密切相关的脊髓背角区域，盐酸布比卡因可阻断 NMDA 受体介导的突触传递 [1]。盐酸布比卡因将半最大激活/失活膜电位移向稍微更负的膜电位，这对通道激活和稳态失活的电压依赖性有影响。 SCN5A 通道对盐酸布比卡因的 IC50 为 2.18±0.16 μM，这表明在非活性状态下有轻微的敏感性[2]。盐酸布比卡因的 IC50 为 16.5 μM，剂量依赖性且可逆地抑制 SK2 通道 [3]。

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作为局部麻醉药，Bupivacaine HCl（1-100 μM）剂量依赖性阻滞神经元细胞的电压门控钠通道，抑制钠离子内流和动作电位传导；10 μM 时在静息膜电位下实现 90% 钠通道阻滞 [1]
- 在人胃癌细胞系（MGC-803、SGC-7901）中，Bupivacaine HCl 抑制细胞增殖，72 小时 IC50 值分别为：MGC-803（1.2 mM）、SGC-7901（1.5 mM）[2]
- 1 mM Bupivacaine HCl 处理 48 小时后，45% 的 MGC-803 细胞和 38% 的 SGC-7901 细胞发生凋亡，表现为 Bax/Bcl-2 比值升高（分别为 3.2 倍和 2.8 倍）、半胱天冬酶 -3 激活（分别为 4.5 倍和 3.9 倍）及 PARP 切割 [2]
- 0.8 mM Bupivacaine HCl 使 MGC-803 和 SGC-7901 细胞的克隆形成率分别降低 65% 和 58%（相对于溶媒对照组）[2]
- Western blot 分析显示，Bupivacaine HCl（0.5-1.5 mM）剂量依赖性下调胃癌细胞中磷酸化 AKT（p-AKT）和磷酸化 mTOR（p-mTOR）：1 mM 时 MGC-803 细胞中 p-AKT 降低 62%，p-mTOR 降低 57% [2]
- 对正常人胃上皮细胞（GES-1）毒性较低：1 mM 浓度处理 72 小时后细胞活力仍 >80% [2]

Bupivacaine 不仅能诱导大鼠肌浆网 (SR) 释放 Ca2+，还能抑制 SR 对 Ca2+ 的摄取，这主要受 SR Ca2+ 三磷酸腺苷酶活性的调节。

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在裸鼠 MGC-803 胃癌异种移植模型中，腹腔注射 Bupivacaine HCl（20 mg/kg，隔日一次，连续 21 天）的肿瘤生长抑制率（TGI）达 56%，肿瘤重量从溶媒组的 1.1 g 降至 0.49 g；肿瘤组织中 TUNEL 阳性凋亡细胞比例达 35%（溶媒组为 7%），p-AKT/p-mTOR 表达降低 [2]
- 在大鼠坐骨神经阻滞模型中，鞘内注射 Bupivacaine HCl（0.5 mg/kg）产生感觉麻醉持续 3.5 小时，运动阻滞持续 2.2 小时 [1]
- 人体临床中，硬膜外注射 Bupivacaine HCl（0.25-0.5% 浓度，10-20 mL）提供术后镇痛 4-8 小时，92% 患者疼痛缓解有效 [1]

电压门控钠通道阻滞实验：培养神经元细胞，采用全细胞膜片钳技术记录。加入系列浓度的 Bupivacaine HCl（1-100 μM），在电压钳模式下记录钠电流。通过比较给药前后的钠电流峰值，计算钠通道阻滞百分比 [1]

细胞活力测定[3]
细胞类型：转染SK2基因的HEK 293细胞（转染细胞命名为SK2细胞）
测试浓度：10、 100, 1000 µM
孵育时间：
实验结果： IC50 值为 16.5 µM。

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抗增殖实验：胃癌细胞（MGC-803、SGC-7901）和正常胃上皮细胞（GES-1）接种于 96 孔板（3×10³ 个细胞 / 孔），用系列浓度的 Bupivacaine HCl（0.1-5 mM）处理 72 小时。MTT 法评估细胞活力，计算 IC50 值 [2]
- 凋亡实验：MGC-803/SGC-7901 细胞用 Bupivacaine HCl（0.5-1.5 mM）处理 48 小时，用膜联蛋白 V-FITC/碘化丙啶染色，流式细胞术分析凋亡率；Western blot 检测 Bax、Bcl-2、切割型半胱天冬酶 -3 及 PARP 表达 [2]
- 克隆形成实验：胃癌细胞用 Bupivacaine HCl（0.4-1.2 mM）处理 24 小时后，接种于 6 孔板（1×10³ 个细胞 / 孔），孵育 14 天。结晶紫染色计数菌落，相对于溶媒对照组计算抑制率 [2]
- 信号通路分析：MGC-803 细胞用 Bupivacaine HCl（0.5-1.5 mM）处理 24 小时，制备细胞裂解液，SDS-PAGE 分离蛋白（AKT、p-AKT、mTOR、p-mTOR），用特异性抗体孵育，密度计量法定量蛋白表达 [2]

Rats

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Gastric cancer xenograft model: 6-8-week-old nude mice were subcutaneously implanted with 5×10⁶ MGC-803 cells. When tumors reached 100-150 mm³, mice were randomized (n=8/group) and treated with: (1) vehicle (DMSO + sterile saline, DMSO ≤5%) via intraperitoneal injection; (2) Bupivacaine HCl (20 mg/kg) via intraperitoneal injection every other day for 21 days. Tumor volume and body weight were measured every 3 days, and tumor tissues were collected for apoptosis and protein expression analysis [2]
- Rat sciatic nerve block model: Adult Sprague-Dawley rats (200-250 g) were anesthetized, and Bupivacaine HCl (0.5 mg/kg, 0.25% concentration) was injected intrathecally near the sciatic nerve. The duration of sensory anesthesia (response to pinprick) and motor block (ability to walk) was recorded [1]
- Bupivacaine HCl was dissolved in sterile saline for animal administration; clinical formulations were sterile injectable solutions (0.25-0.75% concentration) [1][2]

Absorption, Distribution and Excretion
Systemic absorption of local anesthetics depends on the administered dose and concentration, as well as the total amount administered. Other factors affecting the rate of systemic absorption include the route of administration, blood flow at the administration site, and the presence of adrenaline in the anesthetic solution. When bupivacaine reconstituted with meloxicam is administered via infusion, systemic parameters vary after a single dose. In patients undergoing hallux valgus resection, the Cmax of 60 mg bupivacaine was 54 ± 33 ng/mL, the median Tmax was 3 hours, and the AUC∞ was 1718 ± 1211 ng·h/mL. The corresponding values for a 300 mg dose used in hernia repair were 271 ± 147 ng/mL, 18 hours, and 15,524 ± 8921 ng·h/mL, respectively. Finally, the 400 mg dose used in total knee arthroplasty achieved plasma concentrations of 695 ± 411 ng/mL at 21 hours and 38,173 ± 29,400 ng/mL at 21 hours. Only 6% of bupivacaine was excreted unchanged in the urine. After absorption into the bloodstream, bupivacaine hydrochloride exhibits higher plasma protein binding rates than any other local anesthetic; reported binding rates range from 82-96%. Bupivacaine hydrochloride has the lowest placental translocation among all parenteral local anesthetics, and therefore may have the least inhibitory effect on the fetus. Pregnant rats received intravenous infusions of bupivacaine at a rate of 0.33 mg·kg⁻¹·min⁻¹ over 15 minutes. The fetus was delivered at the end of the infusion or 2 or 4 hours after administration. The concentrations of bupivacaine and its metabolites in maternal and fetal blood and tissue samples were determined using capillary gas chromatography-mass spectrometry. The elimination half-life of bupivacaine was 37.7 minutes. The major metabolite was 3'-hydroxybupivacaine. At the end of administration, bupivacaine and 3'-hydroxybupivacaine were detected in all samples. The fetal-to-maternal concentration ratio of bupivacaine in plasma was 0.29, and in the placenta it was 0.63. The highest concentration of bupivacaine was found in the amnion, which was 3 times that of maternal plasma and 11 times that of fetal plasma. Four hours after administration, bupivacaine was undetectable in all maternal and fetal samples, while 3'-hydroxybupivacaine remained in all tissues except fetal plasma and the heart. These data indicate that significant amounts of bupivacaine were absorbed by the placenta, amnion, and myometrium. 3'-hydroxybupivacaine was present in all tissues except fetal plasma and the heart, even when the maternal compound was undetectable. Following tail, epidural, or peripheral nerve block with bupivacaine hydrochloride, peak blood concentrations of bupivacaine are reached within 30 to 45 minutes, subsequently declining to negligible levels over the next 3 to 6 hours. Plasma pharmacokinetic studies following direct intravenous injection of bupivacaine hydrochloride have shown that it conforms to a three-compartment open model. The first compartment represents the rapid intravascular distribution of the drug. The second compartment represents the equilibrium of the drug in highly perfused organs such as the brain, myocardium, lungs, kidneys, and liver. The third compartment represents the equilibrium of the drug with poorly perfused tissues such as muscle and fat. Elimination of the drug from tissue distribution depends primarily on the ability of its binding sites in circulation to transport it to the liver for metabolism. For more complete data on the absorption, distribution, and excretion of bupivacaine (6 in total), please visit the HSDB records page. Metabolites/Metabolites Amide local anesthetics (e.g., bupivacaine) are primarily metabolized in the liver via glucuronide conjugation. The major metabolite of bupivacaine is 2,6-piperidinimide, primarily catalyzed by cytochrome P450 3A4. Pregnant rats received intravenous infusion of bupivacaine at a rate of 0.33 mg·kg⁻¹·min⁻¹ over 15 minutes. The fetus was delivered at the end of the infusion or 2 or 4 hours after administration. Blood and tissue samples were collected from both mother and fetus, and bupivacaine and its metabolites were determined by capillary gas chromatography-mass spectrometry. The elimination half-life of bupivacaine was 37.7 minutes. The major metabolite was 3'-hydroxybupivacaine. Bupivacaine and 3'-hydroxybupivacaine were detected in all samples at the end of administration. The fetal-to-maternal concentration ratio of bupivacaine in plasma was 0.29, and in the placenta it was 0.63. The highest concentration of bupivacaine was found in the amnion: 3 times higher than in maternal plasma and 11 times higher than in fetal plasma. Four hours after administration, bupivacaine was undetectable in all maternal and fetal samples, while 3'-hydroxybupivacaine remained in all tissues except fetal plasma and the heart. These data indicate that significant amounts of bupivacaine were absorbed bilaterally by the placenta, as well as in the amnion and myometrium. Even though the maternal compound was undetectable, 3'-hydroxybupivacaine remained in all tissues except fetal plasma and the heart. Bupivacaine hydrochloride is primarily metabolized to piperidinyl dimethylamine (PPX) via N-dealkylation, a process that may occur in the liver. Bupivacaine is primarily excreted in the urine as a small amount of PPX, the unchanged drug (5%), and other unidentified metabolites. Amide-type local anesthetics (such as bupivacaine) are primarily metabolized in the liver via glucuronide conjugation. The major metabolite of bupivacaine is 2,6-piperidinimide, primarily catalyzed by cytochrome P450 3A4. Elimination pathway: Only 6% of bupivacaine is excreted unchanged in the urine. Half-life: 2.7 hours in adults and 8.1 hours in newborns. The median half-life of bupivacaine in combination with meloxicam for postoperative analgesia is 15-17 hours, depending on the dose and administration site. Pregnant rats received intravenous infusion of bupivacaine at a rate of 0.33 mg·kg⁻¹·min⁻¹ over 15 minutes. The fetus was delivered at the end of the infusion or 2 or 4 hours after administration. Blood and tissue samples were collected from the mother and fetus, and bupivacaine and its metabolites were determined by capillary gas chromatography-mass spectrometry. The elimination half-life of bupivacaine is 37.7 minutes. The elimination half-life of bupivacaine hydrochloride is 1.5-5.5 hours in adults and 8.1 hours in newborns.

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Human pharmacokinetics: After epidural injection of bupivacaine hydrochloride (15 mL 0.5% solution), the peak plasma concentration (Cmax) was 2.8 μg/mL, the terminal half-life (t1/2) was 2.7 hours, and the area under the curve (AUC0-∞) was 12.6 μg·h/mL [1]
- It is mainly metabolized in the liver by cytochrome P450 enzymes (CYP3A4, CYP2C9), of which 70% of the metabolites are excreted in urine and 30% in feces [1]
- The human plasma protein binding rate of bupivacaine is 95-98% at therapeutic concentrations [1]
- The human volume of distribution (Vd) is 1.4 L/kg [1]

Toxicity Summary
Bupivacaine is a cholinesterase, or acetylcholinesterase (AChE) inhibitor. Cholinesterase inhibitors (or "anticholinesterases") inhibit the activity of acetylcholinesterase. Because acetylcholinesterase plays a vital physiological role, chemicals that interfere with its activity are potent neurotoxins; even low doses can cause excessive salivation and lacrimation, followed by muscle spasms and ultimately death. Substances used in nerve gases and many pesticides have been shown to exert their effects by binding to serine residues at the active site of acetylcholinesterase, thereby completely inhibiting the enzyme's activity. Acetylcholinesterase breaks down the neurotransmitter acetylcholine, which is released at the neuromuscular junction, causing muscle or organ relaxation. The mechanism of action of acetylcholinesterase inhibitors is the accumulation and sustained action of acetylcholine, leading to continuous nerve impulse transmission and unstoppable muscle contractions. The most common acetylcholinesterase inhibitors are phosphorus-containing compounds; these compounds act by binding to the enzyme's active site. Its structural requirements are: one phosphorus atom connected to two lipophilic groups, one leaving group (e.g., a halide or thiocyanate), and one terminal oxygen atom.
Toxicity Data
In rhesus monkeys, the mean epileptogenic dose of bupivacaine was 4.4 mg/kg, and the mean arterial plasma concentration was 4.5 mcg/mL.
LD50: 6 to 8 mg/kg (intravenous, mice)
LD50: 38 to 54 mg/kg (subcutaneous, mice)
Interactions
In patients taking monoamine oxidase inhibitors (MAOIs) or tratriptyline or imipramine antidepressants, extreme caution should be exercised when using bupivacaine hydrochloride solutions containing vasoconstrictors (e.g., epinephrine), as this may cause severe, persistent hypertension.
Bupivacaine with epinephrine hydrochloride (1:200,000) or other vasopressors should not be used concomitantly with ergot oxytocin, as this may cause severe, persistent hypertension.

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Non-human toxicity values
Subcutaneous LD50 in mice: 38-54 mg/kg
Intravenous LD50 in mice: 6-8 mg/kg
Central Nervous System (CNS) Toxicity: In humans, plasma concentrations >4 μg/mL may cause dizziness (25% of patients), tinnitus (18%) and seizures (rare, <1%) [1]
-Cardiovascular toxicity: High doses (intravenous >4 mg/kg) may induce bradycardia, hypotension and arrhythmias; the minimum toxic intravenous dose in humans is approximately 2 mg/kg [1]
-Animal toxicity: The LD50 of bupivacaine hydrochloride is 25 mg/kg (intraperitoneal injection in mice) and 10 mg/kg (intravenous injection in rats) [1]
-With bupivacaine hydrochloride (20 mg/kg, every other day for 21 days) Mice treated with (days) showed no significant histopathological abnormalities in the liver, kidneys, or heart; and weight loss was <4% [2]

[1]. Bupivacaine, levobupivacaine and ropivacaine: are they clinically different? Best Pract Res Clin Anaesthesiol. 2005 Jun;19(2):247-68.

[2]. Inhibition of gastric cancer by local anesthetic bupivacaine through multiple mechanisms independent of sodium channel blockade. Biomed Pharmacother. 2018 Jul;103:823-828.

Therapeutic Uses
Bupivacaine hydrochloride is used for infiltration anesthesia, as well as peripheral nerve blocks, sympathetic nerve blocks, and epidural (including tail) block anesthesia. A 0.75% bupivacaine solution (dissolved in 8.25% glucose solution) is used for spinal anesthesia. Bupivacaine is not used for obstetric paracervical blocks or local anesthesia. /See US product label for usage/
Bupivacaine hydrochloride is indicated for local or regional anesthesia or analgesia in surgical, dental and oral surgery, diagnostic and therapeutic procedures, and obstetric procedures. Only 0.25% and 0.5% concentrations of bupivacaine are indicated for obstetric anesthesia. /See US product label for instructions for use/
Drug Warnings Because the use of 0.75% bupivacaine hydrochloride solution for obstetric anesthesia has been associated with cardiac arrest, resuscitation difficulties, or death in obstetric patients, its use in obstetric anesthesia is no longer recommended. Cardiac arrest may occur due to a seizure caused by systemic toxicity, which is clearly due to accidental intravascular injection. Local anesthetics should only be used by clinicians proficient in the diagnosis and management of dose-related toxicities and other acute emergencies that may result from anesthetic blockade, and only after ensuring that oxygen, other resuscitation drugs, cardiopulmonary resuscitation equipment, and the necessary personnel resources for proper management of toxic reactions and related emergencies are readily available. Dose-related toxicities, any cause of inadequate ventilation, and/or changes in sensitivity, if not properly managed in a timely manner, can lead to acidosis, cardiac arrest, or even death. /Local Anesthetics/ Until more data are accumulated regarding the use of this drug in pediatric patients, bupivacaine hydrochloride solution should not be used in children under 12 years of age, and the solution used for spinal anesthesia should not be used in children under 18 years of age. Some commercially available bupivacaine hydrochloride preparations contain sodium metabisulfite, a sulfite that may cause allergic reactions in certain susceptible individuals, including anaphylactic shock and life-threatening or mild asthma attacks. The overall prevalence of sulfite allergy in the general population is unclear but likely low; this sensitivity appears to be more common in asthmatic patients than in non-asthmatic patients. For more complete data on drug warnings for bupivacaine (18 in total), please visit the HSDB records page.

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Pharmacodynamics
Bupivacaine is a widely used local anesthetic. It is usually administered via spinal injection before total hip replacement surgery. It is also frequently injected into the surgical wound site to relieve pain for up to 20 hours post-surgery. It has a longer duration of action compared to other local anesthetics. However, it also has the greatest cardiotoxicity at high doses. This issue has led to the use of other long-acting local anesthetics: ropivacaine and levobupivacaine. Levobupivacaine is a derivative of bupivacaine, specifically its enantiomer. Systemic absorption of local anesthetics can affect the cardiovascular and central nervous systems. Within the range of plasma concentrations achieved at therapeutic doses, changes in cardiac conduction, excitability, refractory period, contractility, and peripheral vascular resistance are minimal. However, toxic blood drug concentrations can inhibit cardiac conduction and excitability, potentially leading to atrioventricular block, ventricular arrhythmias, and cardiac arrest, and sometimes even death. In addition, myocardial contractility is reduced, accompanied by peripheral vasodilation, resulting in decreased cardiac output and arterial blood pressure. Local anesthetics, after systemic absorption, can cause excitation, inhibition, or both of the central nervous system.

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Bupivacaine hydrochloride is an amide-type local anesthetic, clinically applicable to epidural anesthesia, spinal anesthesia, peripheral nerve block anesthesia, and postoperative analgesia[1].
Its core anesthetic mechanism involves reversibly blocking voltage-gated sodium channels on neuronal membranes, thereby preventing sodium ion influx and inhibiting nerve impulse conduction[1].
In addition to its anesthetic effect, it also has antitumor activity against gastric cancer through various sodium channel blockade mechanisms: inducing caspase-dependent apoptosis, inhibiting the PI3K/AKT/mTOR signaling pathway, and inhibiting tumor cell proliferation and colony formation[2]. Compared with short-acting local anesthetics, it has higher potency and longer duration of action, but caution is needed to avoid central nervous system and cardiovascular toxicity [1]. This drug exhibits selective toxicity to gastric cancer cells and minimal damage to normal gastric epithelial cells, suggesting its potential as an adjuvant antitumor drug [2].

C18H28N2O.HCL

324.89

324.196

18010-40-7

Bupivacaine;38396-39-3;Bupivacaine hydrochloride monohydrate;73360-54-0

2474

White to off-white solid powder

423.4ºC at 760 mmHg

107.5 to 108ºC

209.9ºC

4.709

32.34

1

2

5

21

321

0

SIEYLFHKZGLBNX-UHFFFAOYSA-N

InChI=1S/C18H28N2O.ClH/c1-4-5-12-20-13-7-6-11-16(20)18(21)19-17-14(2)9-8-10-15(17)3;/h8-10,16H,4-7,11-13H2,1-3H3,(H,19,21);1H

1-butyl-N-(2,6-dimethylphenyl)piperidine-2-carboxamide hydrochloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮和光照。

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

配方 1 中的溶解度: ≥ 2.5 mg/mL (7.69 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 25.0 mg/mL澄清DMSO储备液加入到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: ≥ 2.5 mg/mL (7.69 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 25.0 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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配方 3 中的溶解度: 13 mg/mL (40.01 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。