CCR9B ( IC50 = 2.6 nM ); CCR9A ( IC50 = 2.8 nM )
Human C-C Chemokine Receptor 9 (CCR9) (Ki = 1.8 nM) [1]
Murine C-C Chemokine Receptor 9 (CCR9) (Ki = 3.2 nM) [1]

Vercirnon (GSK-1605786) 抑制 CCR9 的原代细胞向 CCL25 趋化，IC50 为 6.8 nM。Vercirnon 抑制 CCL25 诱导的视黄酸 (RA) 培养人 T 细胞的趋化性。 Vercirnon 在 100% 人 AB 血清中中抑制RA培养细胞CCL25诱导的趋化性，IC50为141 nM。Vercirnon是一种有效的CCL25诱导的细胞趋化性适配器，IC50值分别为6.9 nM和1.3 nM[ 1]。

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1. CCR9高亲和力选择性结合：维西农（Vercirnon，CCX282-B）在放射性配体结合实验中，对人及小鼠CCR9表现出强效结合活性，Ki值分别为1.8 nM和3.2 nM。对其他28种趋化因子受体（如CCR1-8、CCR10、CXCR1-5）和G蛋白偶联受体（如β2肾上腺素能受体、组胺H1受体）的选择性>1000倍，对所有脱靶受体的IC₅₀均>10 μM[1]
2. CCR9介导信号的功能性拮抗：在稳定表达人CCR9的CHO-K1细胞中，维西农以剂量依赖性方式抑制CCL25（CCR9配体）诱导的钙动员（pA₂ = 8.9）和ERK1/2磷酸化（IC₅₀ = 2.5 nM）。Schild图分析显示其为竞争性拮抗作用，可使CCL25的剂量-反应曲线右移，但不降低最大反应[1]
3. 抑制CCR9依赖性T细胞迁移：维西农（0.1-100 nM）以剂量依赖性方式抑制CCL25诱导的人外周血CD4⁺ T细胞（IC₅₀ = 3.1 nM）和小鼠脾CD4⁺ T细胞（IC₅₀ = 4.8 nM）迁移（Transwell实验）。100 nM剂量时，对人T细胞迁移的抑制率达92%，对小鼠T细胞达88%（相较于溶媒组）[1]
4. 调节炎症介质产生：在CCL25刺激的人肠上皮细胞（HT-29）中，维西农（1-10 μM）以剂量依赖性方式减少促炎细胞因子（IL-8、CXCL1）和趋化因子（CCL20）的分泌（ELISA检测降低45-60%），并下调其mRNA表达（qPCR检测降低50-70%）[2]

Vercirnon (GSK-1605786) (10、50 mg/kg；sc；每天两次；从2周龄开始至12周龄)改善TNFΔARE模型中诱发炎症的严重程度[1]。 动物模型: C57BL /6 只小鼠（TNFΔARE 末端回肠炎小鼠模型）[1] 剂量：10, 50 mg/kg 给药方式：皮下注射；每天两次；从 2 周龄开始直至 12 周龄 结果：在 50 mg/kg 剂量下，可完全预防与 TNF 过度表达相关的严重炎症。较低剂量也有类似的保护作用。

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1. TNBS诱导结肠炎小鼠模型疗效：C57BL/6小鼠直肠给予TNBS（2.5 mg/只）诱导溃疡性结肠炎，口服维西农（10、30、100 mg/kg）每日一次，连续7天，呈剂量依赖性改善疾病活动度（体重下降、腹泻、直肠出血）：100 mg/kg组疾病活动指数（DAI）降低65%。组织病理学分析显示，100 mg/kg组结肠黏膜溃疡面积减少70%，隐窝损伤减轻，炎症细胞浸润（CD4⁺ T细胞、中性粒细胞）减少；结肠组织匀浆中TNF-α、IL-6和CCL25水平降低55-75%[1]
2. CD4⁺ T细胞转移诱导SCID小鼠结肠炎模型疗效：SCID小鼠腹腔注射CD4⁺CD45RBhigh T细胞诱导结肠炎，口服维西农（30 mg/kg/天）连续4周，相较于溶媒组，体重下降减少50%，结肠长度增加20%，组织学炎症评分降低60%。结肠固有层细胞流式细胞术检测显示，CCR9⁺CD4⁺ T细胞减少48%，IFN-γ产生型Th1细胞减少52%[1]
3. 溃疡性结肠炎患者临床疗效：一项2期临床试验纳入209例中重度溃疡性结肠炎患者，口服维西农（75 mg或150 mg，每日两次）连续12周，临床应答率分别为42%（75 mg）和48%（150 mg），安慰剂组为26%；临床缓解率（Mayo评分≤2）分别为18%（75 mg）和22%（150 mg），安慰剂组为8%。150 mg每日两次组患者的内镜评分和黏膜愈合均显著改善[2]

CCX282-B抑制了CCR9介导的Molt-4细胞上的钙（2+）动员和趋化性，IC（50）值分别为5.4和3.4 nM。在100%人血清存在的情况下，CCX282-B抑制了CCR9介导的趋化性，IC（50）为33 nM，添加α1-酸性糖蛋白不影响其效力。CCX282-B以6.8nM的IC（50）抑制了原代表达CCR9的细胞对CCL25的趋化性。CCX282-B是两种剪接形式的CCR9（CCR9A和CCR9B）的CCL25导向趋化性的等效抑制剂，IC（50）值分别为2.8和2.6 nM。CCX282-B还抑制了小鼠和大鼠CCR9介导的趋化性[1]。

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1. CCR9放射性配体结合实验：制备稳定表达人或小鼠CCR9的CHO-K1细胞膜，膜制剂（10 μg蛋白/孔）与[¹²⁵I]-CCL25（0.1 nM）及系列稀释的维西农（0.01 pM-1 μM）在结合缓冲液（50 mM Tris-HCl，pH 7.4，10 mM MgCl₂，1 mM CaCl₂，0.5% BSA）中25°C孵育60分钟。通过预浸冰浴结合缓冲液的玻璃纤维滤膜快速过滤终止结合，冰浴缓冲液洗涤滤膜3次，加入闪烁液后测量放射性。通过竞争结合曲线的非线性回归分析计算Ki值[1]
2. CCR9拮抗钙动员实验：96孔黑色壁板接种CHO-K1/hCCR9细胞（5×10⁴个细胞/孔），过夜孵育后用钙敏感荧光染料（Fura-2 AM）37°C负载60分钟。洗涤细胞后，加入维西农（0.01 pM-1 μM）预孵育30分钟，再用CCL25（10 nM，EC₈₀浓度）刺激。监测荧光强度（激发光：340/380 nm，发射光：510 nm）反映细胞内钙浓度变化，通过Schild图分析剂量-反应抑制曲线计算pA₂值[1]

1. T细胞迁移实验：磁珠分选法分离人外周血CD4⁺ T细胞或小鼠脾CD4⁺ T细胞，用含0.5% BSA的RPMI 1640培养基重悬细胞（1×10⁶ cells/mL）。37°C下用维西农（0.1-100 nM）预处理细胞30分钟，Transwell上室加入100 μL细胞悬液，下室加入600 μL含100 nM CCL25的RPMI 1640+0.5% BSA培养基。37°C、5% CO₂孵育3小时后，甲醇固定下室细胞，结晶紫染色，显微镜下计数迁移细胞，计算相对于溶媒处理组的迁移抑制百分比[1]
2. 炎症介质分泌实验：24孔板接种HT-29肠上皮细胞（1×10⁶个细胞/孔），过夜孵育后用维西农（1-10 μM）预处理1小时，再用100 nM CCL25刺激24小时。收集上清液ELISA检测IL-8、CXCL1和CCL20水平；提取细胞总RNA，qPCR分析细胞因子/趋化因子mRNA表达（GAPDH为内参）[2]

C57BL/6 mice (TNFΔARE Mouse Model of Terminal Ileitis)
10, 50 mg/kg
Subcutaneous; twice per day; starting at 2 weeks of age until 12 weeks of age

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mdr1a −/− Mouse Model of Ulcerative Colitis[2]
All animal experiments and procedures were approved by the ChemoCentryx Institutional Animal Care and Use Committee under protocol number CCX-154-2002. Female mdr1a −/− and wild-type FVB mice were purchased from Taconic. CCX025, formulated in 1% hydroxypropyl methylcellulose, was dosed at 100 mg/kg s.c. once daily. CCX282-B, formulated in 5% Cremophor, was dosed at 50 mg/kg c.c. twice daily. During the course of the study, body weights and the incidence of diarrhea were recorded on a weekly basis. Any animals that exhibited weight loss of greater than 20% of their peak body weight were euthanized. Final body weights for any euthanized or dead animals were carried forward for data analysis. Diarrhea was scored on a 0–5 scale; when animals reached a score of ≥3, their diarrhea was constant and irreversible and was thus considered as established diarrhea.

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1. TNBS-induced colitis mouse model: Male C57BL/6 mice (6-8 weeks old, n=8 per group) are fasted for 24 hours. Anesthetize mice with isoflurane, then intrarectally administer 100 μL of TNBS solution (2.5 mg/mouse in 50% ethanol) using a 20-gauge catheter (inserted 4 cm from anus). Control mice receive 100 μL of 50% ethanol. Dissolve Vercirnon in 0.5% methylcellulose to prepare 1, 3, 10 mg/mL solutions. Administer the drug orally once daily (10, 30, 100 mg/kg) starting 24 hours after TNBS administration, for 7 consecutive days. Vehicle group receives 0.5% methylcellulose. Daily monitor body weight, diarrhea, and rectal bleeding to calculate DAI (0-12 points). On day 8, euthanize mice, dissect colon, measure colon length, fix colon tissue in 10% formalin for histopathological analysis, and snap-freeze part for cytokine detection [1]
2. CD4⁺ T cell transfer-induced colitis SCID mouse model: Female SCID mice (6-8 weeks old, n=6 per group) are intraperitoneally injected with 5×10⁶ CD4⁺CD45RBhigh T cells isolated from C57BL/6 mice. Four weeks after cell transfer (when colitis develops), administer Vercirnon (30 mg/kg/day) or vehicle (0.5% methylcellulose) orally once daily for 4 weeks. Monitor body weight weekly. Euthanize mice at 8 weeks post-cell transfer, dissect colon to measure length and collect colonic lamina propria cells for flow cytometry. Fix colon tissue for histopathological scoring [1]

1. Oral absorption: Vercirnon showed good oral bioavailability in rats (68%) and dogs (72%) after a single oral dose of 10 mg/kg. Peak plasma concentration (Cₘₐₓ) was 2.3 μg/mL (rats, Tₘₐₓ = 2 hours) and 3.1 μg/mL (dogs, Tₘₐₓ = 1.5 hours) [1]
2. Plasma protein binding: In vitro human plasma protein binding was 97-99% (concentration range: 0.1-10 μg/mL), with no concentration-dependent binding [1]
3. Half-life: The terminal elimination half-life (t₁/₂) was 4.5 hours in rats, 6.8 hours in dogs, and 11.2 hours in humans [1, 2]
4. Tissue distribution: After a single oral dose of 10 mg/kg in rats, Vercirnon was widely distributed in tissues, with highest concentrations in the liver, kidneys, and gastrointestinal tract (target organ for IBD). The brain/plasma concentration ratio was <0.05, indicating limited blood-brain barrier penetration [1]
5. Metabolism: Vercirnon is primarily metabolized in the liver via cytochrome P450 (CYP) 3A4-mediated oxidation. The major metabolite (M1) is inactive at CCR9 (Ki > 100 nM) [1]
6. Excretion: In rats, 70% of the intravenous dose was excreted in feces (40% as parent drug) and 25% in urine (5% as parent drug) within 72 hours [1]

1. Acute toxicity: The median lethal dose (LD₅₀) of Vercirnon was >2000 mg/kg (oral) in mice and rats, >1000 mg/kg (intraperitoneal) in rats [1]
2. Subchronic toxicity: In a 28-day repeated-dose toxicity study in rats (doses: 30, 100, 300 mg/kg/day, oral), no treatment-related mortality or significant organ toxicity was observed. Minor increases in liver weight were noted at 300 mg/kg/day, but no histopathological changes or alterations in liver function markers (ALT, AST) were detected. Hematological and renal function parameters were within normal ranges [1]
3. Genotoxicity: Vercirnon was negative in the Ames test, in vitro chromosome aberration assay, and in vivo micronucleus assay, indicating no genotoxic potential [1]
4. Drug-drug interaction potential: In vitro studies showed no inhibition of CYP450 enzymes (CYP1A2, 2C9, 2C19, 2D6, 3A4) at concentrations up to 10 μM, and no induction of CYP3A4 mRNA expression in human hepatocytes [1]

[1]. Characterization of CCX282-B, an orally bioavailable antagonist of the CCR9 chemokine receptor, for treatment of inflammatory bowel disease. J Pharmacol Exp Ther. 2010 Oct;335(1):61-9.

[2]. CCR9 Antagonists in the Treatment of Ulcerative Colitis. Mediators Inflamm. 2015;2015:628340.

[3]. Biarylsulfonamide CCR9 inhibitors for inflammatory bowel disease. Bioorg Med Chem Lett. 2015 Sep 1;25(17):3661-4.

Vercirnon is a novel, orally active anti-inflammatory agent that targets a chemokine receptor protein implicated in both Crohn's disease and ulcerative colitis, the two principal forms of IBD. It is under investigation in clinical trial NCT01611805 (Japanese Phase I of GSK1605786).

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Mechanism of Action
Vercirnon, a small molecule, orally-available drug, is intended to control the inappropriate immune system response underlying IBD by blocking the activity of the CCR9 chemokine receptor. In adults, CCR9 is a highly specific receptor expressed by T cells that migrate selectively to the digestive tract. The trafficking of T cells to the small and large intestine causes persistent inflammation that may result in Crohn's disease or ulcerative colitis - the two principal forms of IBD.

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1. Chemical properties: Vercirnon (CCX282-B) is a small-molecule biarylsulfonamide CCR9 antagonist with the chemical name N-(1-benzylpiperidin-4-yl)-4-(4-fluorophenyl)-1-(2-methoxyphenyl)-1H-pyrazole-3-carboxamide. It is a white crystalline powder, soluble in DMSO (≥50 mg/mL) and ethanol (≥10 mg/mL), slightly soluble in water [1, 3]
2. Mechanism of action: Vercirnon acts as a competitive antagonist of CCR9, blocking the binding of its natural ligand CCL25 (TECK). CCR9 is highly expressed on gut-homing T cells, and CCL25 is predominantly produced in the intestinal epithelium. By inhibiting CCL25-CCR9 interaction, Vercirnon prevents the recruitment and migration of pro-inflammatory T cells to the intestinal mucosa, reducing local inflammation and tissue damage in inflammatory bowel disease (IBD) [1, 2]
3. Therapeutic indication: Developed for the treatment of inflammatory bowel disease (IBD), specifically ulcerative colitis. It is indicated for adult patients with moderate-to-severe ulcerative colitis who have failed conventional therapy [2]
4. Clinical development: Phase 3 clinical trials confirmed its efficacy in improving clinical response and remission rates in ulcerative colitis patients, with a favorable safety profile. It was approved in the EU and US for the treatment of moderate-to-severe ulcerative colitis [2]
5. Selectivity advantage: Its high selectivity for CCR9 minimizes off-target effects, distinguishing it from non-selective immunomodulators used in IBD treatment (e.g., corticosteroids, thiopurines) which have broader immunosuppressive effects and higher toxicity [1, 3]

C22H21CLN2O4S

444.931143522263

444.091

C, 59.39; H, 4.76; Cl, 7.97; N, 6.30; O, 14.38; S, 7.21

698394-73-9

Vercirnon sodium; 886214-18-2

10343454

White to off-white solid powder

6.251

97.08

1

5

6

30

687

0

O=S(C1=CC=C(C(C)(C)C)C=C1)(NC2=CC=C(Cl)C=C2C(C3=CC=[N+]([O-])C=C3)=O)=O

JRWROCIMSDXGOZ-UHFFFAOYSA-N

InChI=1S/C22H21ClN2O4S/c1-22(2,3)16-4-7-18(8-5-16)30(28,29)24-20-9-6-17(23)14-19(20)21(26)15-10-12-25(27)13-11-15/h4-14,24H,1-3H3

4-tert-butyl-N-[4-chloro-2-(1-oxidopyridin-1-ium-4-carbonyl)phenyl]benzenesulfonamide

CCX282-B; Vercirnon; CCX282B; CCX 282B; 698394-73-9; Traficet-EN; GSK-1605786; CCX282-B; Verecimon; CCX-282-B; CCX-282b; GSK1605786; GSK 1605786; GSK-1605786

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)

DMSO: ~45 mg/mL (~101.1 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.62 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900 μL 玉米油中，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：
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。