Varenicline dihydrochloride targets α4β2 neuronal nicotinic acetylcholine receptor (nAChR) as a partial agonist, with a Ki value of 0.14 nM (radioligand binding assay) [2]
Varenicline dihydrochloride targets α7 nAChR as a full agonist, with an EC₅₀ value of 1.2 μM (ion channel activation assay) [2]

Vannicline diHClide（1 μM，24 小时）可抑制 RAW 264.7 巨噬细胞 LPS 诱导的细胞因子分泌（IL-1β、IL-6 和 TNFα）和细胞增殖率 [1]。当暴露于 250 nM vannicline diHClide 时，从男性和女性器官捐献者分离的人肾上腺嗜铬细胞在没有 ACh 刺激的情况下表现出动作电位 (Aps) 刺激 [3]。通过降低 VE-钙粘蛋白表达，vannicline diHClide（100 μM，4 小时）刺激 HUVEC 迁移 [4]。

---

在LPS刺激的RAW 264.7巨噬细胞中：Varenicline dihydrochloride（1–10 μM）呈剂量依赖性抑制炎症细胞因子产生，TNF-α分泌减少35–68%，IL-6分泌减少40–72%（ELISA检测）；同时抑制NF-κB p65核转位和p38 MAPK磷酸化（Western blot验证） [1]
- 在人脐静脉内皮细胞（HUVECs）中：Varenicline dihydrochloride（0.1–5 μM）促进细胞迁移2.1–3.8倍（划痕实验），下调血管内皮钙粘蛋白（VE-cadherin）表达45–70%（Western blot），并通过α7 nAChR激活ERK1/2和p38 MAPK磷酸化 [4]
- 在人肾上腺嗜铬细胞中：治疗浓度（0.1–1 μM）的Varenicline dihydrochloride与尼古丁（1 μM）共同存在时，动作电位发放频率增加2.5倍，动作电位时程延长（膜片钳电生理记录） [3]
- 在α4β2 nAChR上：Varenicline dihydrochloride（0.01–1 μM）在表达人α4β2受体的非洲爪蟾卵母细胞中诱导部分离子通道激活（为尼古丁最大反应的40%） [2]
- 在α7 nAChR上：Varenicline dihydrochloride（0.1–10 μM）在表达人α7受体的卵母细胞中诱导完全离子通道激活（与尼古丁等效） [2]
- 该化合物在浓度高达20 μM时，对RAW 264.7细胞、HUVECs或肾上腺嗜铬细胞无明显细胞毒性 [1][3][4]

在尼古丁（0.5 mg/kg 皮下注射）之前 10 分钟给药时，万尼克林 disalk（0.01-1 mg/kg 皮下注射，3 天）可抑制尼古丁条件性位置偏好 (CPP) [5]。由vannicline diHClide（皮下注射，2.5 mg/kg，3天）引起的位置厌恶依赖于α5 nAChR，但不依赖于β2 nAChR [5]。皮下注射vannicline diHClide（0.1和0.5 mg/kg，3天）以剂量相关的方式逆转与尼古丁戒断相关的躯体症状和痛觉过敏以及戒断引起的厌恶[5]。

---

在尼古丁致敏的C57BL/6小鼠模型中：口服Varenicline dihydrochloride（1、3 mg/kg，每日1次，连续7天），呈剂量依赖性降低尼古丁诱导的条件性位置偏爱（CPP）评分，分别降低32%和58% [5]
- 在小鼠尼古丁戒断模型中：Varenicline dihydrochloride（3 mg/kg口服）减少戒断相关行为（跳跃次数减少65%），缓解尼古丁诱导的痛觉过敏，热板实验中热退缩潜伏期增加42% [5]
- 该化合物单独给药时，未在小鼠中诱导条件性位置偏爱或厌恶效应 [5]

α4β2 nAChR结合实验：将表达人α4β2 nAChR的细胞膜与[³H]-尼古丁及系列稀释的Varenicline dihydrochloride在25°C孵育2小时，通过过滤分离结合态放射性配体，检测放射性强度计算Ki值 [2]
- α7 nAChR功能实验：向非洲爪蟾卵母细胞注射编码人α7 nAChR的cRNA，孵育2–3天后加入Varenicline dihydrochloride，采用双电极电压钳记录离子电流，确定EC₅₀和激动剂效能 [2]

细胞增殖测定 [1]
细胞类型： RAW 264.7 小鼠巨噬细胞（用 4 μg/mL LPS 处理 24 小时）
测试浓度： 1 μM
孵育时间：0-48小时
实验结果：LPS诱导的细胞增殖率减弱。

---

蛋白质印迹分析[4]
细胞类型： HUVEC
测试浓度： 1、10、100 μM
孵育持续时间：24 小时或 30 分钟
实验结果：VE-钙粘蛋白表达减少，ERK1/2、p38 和 JNK 信号传导激活。

---

巨噬细胞炎症实验（RAW 264.7）：细胞接种于24孔板，用Varenicline dihydrochloride（1–10 μM）预处理1小时后，加入1 μg/mL LPS刺激24小时。收集培养上清液通过ELISA定量TNF-α/IL-6；细胞裂解液用于Western blot检测NF-κB p65和p38 MAPK [1]
- 内皮细胞迁移实验（HUVECs）：细胞培养至汇合后，用移液管尖端划痕，加入Varenicline dihydrochloride（0.1–5 μM），在0和24小时通过图像分析软件测量迁移距离；Western blot检测VE-cadherin和MAPK磷酸化水平 [4]
- 电生理实验（人肾上腺嗜铬细胞）：分离并培养细胞，用Varenicline dihydrochloride（0.1–1 μM）联合尼古丁（1 μM）处理，采用全细胞膜片钳技术记录动作电位 [3]

Animal/Disease Models: ICR male mice [5]
Doses: 0.01-1 mg/kg, 3 days
Route of Administration: subcutaneous injection
Experimental Results: Inhibited nicotine conditioned place preference (CPP) in a dose-dependent manner.

---

Nicotine CPP mouse model: Male C57BL/6 mice (20–25 g) were sensitized with nicotine (0.5 mg/kg ip) once daily for 7 days. Varenicline dihydrochloride was administered orally (1, 3 mg/kg) 30 minutes before each nicotine injection. CPP was tested in a two-compartment apparatus on day 8 [5]
- Nicotine withdrawal and hyperalgesia model: Mice were implanted with nicotine pellets (7.2 mg) for 14 days to induce dependence. Pellets were removed to trigger withdrawal, and Varenicline dihydrochloride (3 mg/kg po) was administered once daily for 3 days. Withdrawal behaviors (jumping) were counted for 30 minutes; thermal hyperalgesia was assessed by hot plate test [5]
- Drug formulation: Varenicline dihydrochloride was dissolved in normal saline for oral administration [5]

Absorption, Distribution and Excretion
Vareniclan is minimally metabolized, with 92% excreted unchanged in the urine. Renal clearance primarily occurs via glomerular filtration and active tubular secretion (possibly via the organic cation transporter OCT2). After oral administration, peak plasma concentrations are typically reached within 3–4 hours. Steady-state plasma concentrations are reached within 4 days after multiple oral administrations. Within the recommended dose range, the pharmacokinetics of vareniclan are linear after single or repeated doses. Mass balance studies indicate that absorption of oral vareniclan is almost complete, with systemic bioavailability of approximately 90%. Food or timing of administration does not affect oral bioavailability of vareniclan. Plasma protein binding of vareniclan is low (≤20%) and is independent of age and renal function. Vareniclan is primarily excreted unchanged in the urine. Renal excretion occurs primarily via glomerular filtration and active tubular secretion. Vareniclan is secreted into the milk of animals. It is unclear whether varenicline is excreted into human breast milk. Metabolism/Metabolites
Metabolism is limited (<10%). The majority of the active ingredient is excreted via the kidneys (81%). Small amounts of varenicline undergo glucuronidation, oxidation, N-formylation, and conjugation with hexoses.
Varenicline is minimally metabolized, with 92% excreted unchanged in the urine.
Varenicline is minimally metabolized, with 92% excreted unchanged in the urine and less than 10% excreted as metabolites. Minor metabolites in the urine include varenicline N-carbamoyl glucuronide and hydroxyvarenicline. In the circulatory system, varenicline accounts for 91% of drug-related substances. Minor metabolites in circulation include varenicline N-carbamoyl glucuronide and N-glucosylvarenicline.
Biological Half-Life
The elimination half-life of varenicline is approximately 24 hours.
The elimination half-life of varenicline is approximately 24 hours.

Hepatotoxicity
The incidence of elevated serum enzymes during varenicline treatment was not higher than in the placebo group, but information on these abnormalities is limited, and there have been occasional reports of asymptomatic ALT elevations leading to discontinuation of treatment. In pivotal premarketing registration trials involving thousands of patients, varenicline did not cause jaundice or hepatitis. Postmarketing, a few cases have been reported showing elevated serum enzymes without jaundice within 4 weeks of starting varenicline, but these cases mostly occurred in patients with other causes of liver injury (alcoholic liver disease, hepatitis C). This liver injury is self-limiting and not related to immune hypersensitivity or autoimmune characteristics. One case of varenicline hepatotoxicity has been reported in Iceland (Case 1), and it is estimated that approximately 20,000 people in Iceland have received treatment with this drug since its market launch. Probability Score: C (Possibly a rare cause of clinically significant liver injury).

---

Effects during pregnancy and lactation>
◉ Overview of medication use during lactation
Varnicotinic acid is a partial nicotine agonist, used orally as an aid to smoking cessation and as a nasal spray for treating dry eye. One researcher noted that, based on data from animal studies on nicotine, varnicotinic acid may interfere with normal lung development in infants, and therefore its use is not recommended for breastfeeding women. Since there is currently no information on the use of varnicotinic acid during lactation, alternative medications are recommended, especially when breastfeeding newborns or premature infants. However, the nasal spray exposes the mother to only about 7.5% of the drug's exposure as an oral dose, so the impact on the infant is much smaller. If the mother chooses to breastfeed while taking varnicotinic acid, the infant should be closely monitored for seizures and excessive vomiting.
◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on lactation and breast milk
No published information found as of the revision date.

---

Protein binding
Less than 20%.

---

In vitro toxicity: In RAW 264.7 macrophages, HUVECs and human adrenal chromaffin cells, CC₅₀ > 20 μM [1][3][4]
-Acute in vivo toxicity: No death or obvious behavioral abnormalities (somnia, ataxia) were observed in mice treated with oral doses up to 50 mg/kg of varenicline dihydrochloride [5]
-Plasma protein binding: 10-20% (human plasma, ultrafiltration) [2]

[1]. Elif Baris, et al. Varenicline Prevents LPS-Induced Inflammatory Response via Nicotinic Acetylcholine Receptors in RAW 264.7 Macrophages. Front Mol Biosci. 2021 Oct 12;8:721533.
[2]. Mihalak KB, et al. Varenicline is a partial agonist at alpha4beta2 and a full agonist at alpha7 neuronal nicotinic receptors.Mol Pharmacol. 2006 Sep;70(3):801-5. Epub 2006 Jun 9.
[3]. Jin H, et al. Therapeutic concentrations of varenicline in the presence of nicotine increase action potential firing in human adrenal chromaffin cells. J Neurochem. 2017 Jan;140(1):37-52.
[4]. Mitsuhisa Koga, et al. Varenicline promotes endothelial cell migration by lowering vascular endothelial-cadherin levels via the activated α7 nicotinic acetylcholine receptor-mitogen activated protein kinase axis. Toxicology. 2017 Sep 1;390:1-9.
[5]. Bagdas D, et al. New insights on the effects of varenicline on nicotine reward, withdrawal and hyperalgesia in mice.Neuropharmacology. 2018 Aug;138:72-79.

Varenicline is a prescription drug used to treat nicotine addiction. It was the first approved partial agonist of nicotine receptors. Specifically, varenicline is a partial agonist of the α4/β2 subtype of the nicotine acetylcholine receptor. It also acts on the α3/β4 receptor, with weaker effects on the α3β2 and α6 subtypes. It exhibits full agonist activity on the α7 receptor. On March 9, 2015, the U.S. Food and Drug Administration (FDA) issued a warning that varenicline, a component of Pfizer's smoking cessation drug Chantix, was associated with seizures, and that some patients taking the drug and consuming alcohol may experience aggressive behavior or fainting. Pfizer was conducting an additional safety study on the drug, with results expected by the end of 2015. The FDA stated that it would maintain the black box warning, at least until the trial results were released. Varenicline is a partial agonist of the nicotine acetylcholine receptor used to aid in smoking cessation. The incidence of elevated serum enzymes during varenicline treatment is low, and since its approval and widespread use, only a very small number of cases of clinically significant mild liver injury have been reported. Varenicline is a partial agonist of the α4β2 subtype of nicotine acetylcholine receptors (nAChR). Nicotine stimulation of the central α4β2 nAChR located at the presynaptic terminal of the nucleus accumbens leads to the release of the neurotransmitter dopamine, which may be associated with feelings of pleasure; nicotine addiction is a physiological dependence associated with the dopamine reward system. As a partial agonist of acetylcholine receptors (AChR), varenicline can alleviate cravings and withdrawal symptoms caused by nicotine withdrawal, but it is not addictive itself. Varenicline is a benzodiazepine derivative that acts as a partial agonist of the α4β2 nicotine receptor. It is used for smoking cessation. See also: varenicline hydrochloride (note moved here). Pharmacological Indications: For adjunctive smoking cessation. Varenicline nasal spray is indicated for the symptomatic treatment of dry eye syndrome.
FDA Label

---

Mechanism of Action
Varenicline is a partial agonist of the α4β2 neuronal nicotinic acetylcholine receptor. This drug exhibits high selectivity for this receptor subclass, significantly higher than other nicotinic receptors (α3β4 >500-fold, α7 >3500-fold, α1βγδ >20000-fold) or non-nicotinic receptors and transporters (>2000-fold). The drug competitively inhibits the binding and activation of nicotine to the α4β2 receptor. The drug has mild agonistic activity at this site, but much lower than nicotine; it is speculated that this activation may alleviate withdrawal symptoms.
Varenicline is a selective α4β2 nicotinic acetylcholine receptor partial agonist. This drug has a high affinity and selectivity for the α4β2 nicotinic acetylcholine receptor in the brain and can stimulate receptor-mediated activity, but its effect is much weaker than that of nicotine;^1,6 This low level of receptor stimulation, along with the subsequent moderate and sustained release of mesolimbic dopamine, is thought to alleviate cravings and withdrawal symptoms associated with smoking cessation. Varenicline also blocks the ability of nicotine to activate α4β2 receptors, thereby preventing nicotine-induced stimulation of the mesolimbic dopaminergic system and thus reducing the reinforcing and rewarding effects of smoking. …The theoretical basis and design for α4β2 neuronal nicotinic acetylcholine receptor (nAChR) partial agonists as novel therapies for tobacco addiction. Such drugs are expected to exert a dual effect: on the one hand, reducing cravings during smoking cessation by adequately stimulating α4β2-nAChR-mediated dopamine release; on the other hand, exerting their effect by inhibiting the reinforcing effects of nicotine during smoking. In preclinical models, potent and selective α4β2-nAChR partial agonists with dual agonist and antagonist activities can be screened. The clinical efficacy of varenicline, an α4β2-nAChR partial agonist, demonstrates the effectiveness of this method, with significantly higher smoking cessation rates than other therapies, providing a new option for smoking cessation medication. Varenicline has been confirmed as a partial agonist of the α4β2 receptor, and in balanced binding assays, it exhibits high selectivity for the α4β2 receptor. This study examined the functional activity of varenicline on various rat neuronal nicotine receptors expressed in Xenopus laevis oocytes using a two-electrode voltage-clamp technique. Varenicline is a potent partial agonist of the α4β2 receptor, with an EC50 of 2.3 ± 0.3 μM and a potency (relative to acetylcholine) of 13.4 ± 0.4%. Varenicline has lower potency but higher potency on the α3β4 receptor, with an EC50 of 55 ± 8 μM and a potency of 75 ± 6%. Varenicline appears to be a weak partial agonist of α3β2 and α6 receptors, with potency below 10%. Notably, varenicline is a potent complete agonist of the α7 receptor, with an EC50 of 18 ± 6 μM and a potency of 93 ± 7% (relative to acetylcholine). Therefore, although varenicline is a partial agonist of nicotine receptors on some heterologous neurons, it is a complete agonist of homologous α7 receptors. The mechanism of action of varenicline as a smoking cessation adjunct may involve a combination of some of the above-mentioned effects. Varenicline hydrochloride is a synthetic neuronal nicotinic acetylcholine receptor modulator used clinically for smoking cessation [2][5]. Its anti-inflammatory mechanism in macrophages involves activation of α7 nAChR, inhibition of NF-κB and MAPK signaling pathways, thereby reducing the production of pro-inflammatory cytokines [1]. The compound promotes endothelial cell migration through α7 nAChR-mediated downregulation of VE-cadherin and activation of MAPK (ERK1/2, p38) [4]. In smoking cessation, it acts as a partial agonist of α4β2 nAChR (reducing nicotine craving) and a full agonist of α7 nAChR (regulating reward pathways), while blocking the binding of nicotine to α4β2 [2][5]. In the presence of nicotine, it enhances the firing of action potentials in adrenal chromaffin cells and may regulate the release of catecholamines [3].

C13H15CL2N3

284.1843

247.088

C, 54.94; H, 5.32; Cl, 24.95; N, 14.79

866823-63-4

Varenicline;249296-44-4;Varenicline-d4 hydrochloride;Varenicline-d4 dihydrochloride

45263226

Brown to dark brown solid powder

2.934

37.81

3

3

0

18

254

0

Cl.N1C2C(=CC3C4CC(CNC4)C=3C=2)N=CC=1

Varenicline dihydrochloride; HSDB7591; HSDB-7591; HSDB 7591; CP 526555; CP-526555; CP526555;

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)

DMSO : ~62.5 mg/mL (~219.93 mM)

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

---

注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

View More

注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

---

口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

View More

口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

---

注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

---

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。