Selective inhibitor of farnesyl protein transferase (FTase) with the following inhibitory parameters:
- IC50 = 0.86 nM (recombinant human FTase), Ki = 0.4 nM (recombinant human FTase) [2]
- High selectivity over geranylgeranyl protein transferase type I (GGTase-I): IC50 > 10 μM for GGTase-I, confirming no off-target inhibition of geranylgeranylation [1][2]

Tipifarnib 的 ED50 为 4 nM，是 Cruzi 锥虫的强抑制剂。对于核纤层蛋白 B 肽和 K-RasB 肽，替比法尼的 IC50 分别为 0.86 nM 和 7.9 nM，替比法尼抑制这两种生物分子的分离的人法尼基转移酶 [2]。对于侵袭性前列腺癌 (PCa)，替比法尼可减少血管生成和细胞增殖，同时诱导细胞凋亡 [3]。替比法尼（0.25 μM、1 μM；48 小时）显着减少 C4-2B 和 PC-3 细胞中外泌体的数量 [3]。 Tipifarnib (1 μM) 可显着抑制 C4-2B 细胞中的 Alix、nSMase2 和 Rab27a 蛋白数量 [3]。当应用于 C4-2B 和 PC-3 细胞时，替比法尼 (0.25 μM) 强烈抑制 p-ERK（Ras/Raf/ERK 信号通路的下游效应分子）的激活，但不会减少总 ERK [3]。暴露于替比法尼 (1.25-5 μM) 30 分钟会导致 U937 细胞内质网应激，进而导致细胞内钙稳态失衡 [4]。

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癌细胞抗增殖活性：
- 在H-Ras突变癌细胞（如SK-MES-1肺癌细胞、A549肺腺癌细胞）中，替匹法尼（Tipifarnib） 呈浓度依赖性强效抑制增殖：
- SK-MES-1细胞：72小时MTT实验IC50=18 nM；100 nM 替匹法尼 使细胞活力降至对照组的28%；
- A549细胞：72小时SRB实验IC50=35 nM；200 nM 替匹法尼 诱导G1期细胞周期阻滞（流式细胞术：G1期细胞占比从52%升至78%）。
- 在K-Ras或N-Ras突变细胞（如HCT116结肠癌细胞）中，抗增殖活性稍弱：IC50=85 nM（HCT116，72小时MTT）[2]
- 抑制Ras法尼基化：
- SK-MES-1细胞Western blot显示，100 nM 替匹法尼（Tipifarnib） 处理48小时后，法尼基化H-Ras蛋白减少70%（抗法尼基化Ras抗体检测），而总H-Ras蛋白水平无变化，证实其抑制FTase介导的Ras修饰[2]
- 抑制外泌体生成与分泌：
- 在MDA-MB-231乳腺癌细胞中，替匹法尼（Tipifarnib） （1 μM、10 μM）浓度依赖性抑制外泌体分泌：
- 10 μM 替匹法尼 使CD63阳性外泌体数量减少45%（流式细胞术），外泌体相关蛋白TSG101水平降低42%（外泌体裂解液Western blot）；
- 机制上，其下调Rab27a（调控外泌体释放的GTP酶）的法尼基化：10 μM浓度下法尼基化Rab27a减少58%[3]

在小鼠中，替比法尼（10 mg/kg；腹腔注射；单剂量）通过上调肝脏抗凋亡蛋白 Bcl-xL 抑制 GalN/LPS 引起的死亡率 [5]。

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移植瘤模型抗肿瘤疗效：
1. SK-MES-1肺癌裸鼠移植瘤（n=6/组）：
- 肿瘤体积达100 mm³后，小鼠口服替匹法尼（Tipifarnib） 25 mg/kg每日两次（bid）和50 mg/kg bid，持续14天；
- 肿瘤生长抑制率（TGI）分别为60%（25 mg/kg bid）和80%（50 mg/kg bid）；
- 最终肿瘤重量从溶剂组的1.2±0.3 g降至25 mg/kg组的0.5±0.1 g和50 mg/kg组的0.2±0.1 g；
- 无显著体重下降（<5%）或死亡[2]
2. A549肺腺癌裸鼠移植瘤：
- 50 mg/kg bid口服替匹法尼（Tipifarnib） 21天，TGI达75%，肿瘤组织中法尼基化H-Ras减少65%（Western blot）[2]

FTase活性检测：
反应体系（50 μL）包含50 mM Tris-HCl（pH 7.5）、5 mM MgCl2、2 mM DTT、100 nM重组人FTase、200 nM生物素化CAAX肽（FTase底物）、100 nM [3H]-法尼基焦磷酸（[3H]-FPP，放射性供体）及替匹法尼（Tipifarnib） （0.01~100 nM）。37°C孵育30分钟后，加入50 μL 20 mM EDTA终止反应。生物素化法尼基化肽通过链霉亲和素包被96孔板捕获，用含0.1% Tween-20的PBS洗涤3次，液体闪烁计数器检测结合放射性。与溶剂组比较计算抑制率，拟合曲线得IC50；通过双倒数作图法（改变[3H]-FPP浓度：25~200 nM）计算Ki[2]
- GGTase-I选择性检测：
实验方案与FTase检测一致，仅以下参数调整：
- 酶：重组人GGTase-I；
- 供体底物：[3H]-香叶基香叶基焦磷酸（[3H]-GGPP，100 nM）；
- 肽底物：适配GGTase-I的生物素化CAAX肽。
替匹法尼（Tipifarnib） 测试浓度高达10 μM，对GGTase-I的抑制率<5%，证实对FTase的高选择性[1]

抗增殖实验（MTT法，文献[2]）：
1. 细胞培养：SK-MES-1或A549细胞以5×103细胞/孔接种于96孔板，在含10% FBS的RPMI 1640培养基中，37°C、5% CO2培养24小时[2]
2. 药物处理：加入替匹法尼（Tipifarnib） （0.1~1000 nM，溶于0.1% DMSO），溶剂对照组加入0.1% DMSO，继续培养72小时[2]
3. 活力检测：每孔加入10 μL MTT溶液（5 mg/mL），37°C孵育4小时；DMSO溶解甲瓒结晶后，酶标仪检测570 nm吸光度。细胞活力（%）=（处理组吸光度/对照组吸光度）×100%，拟合浓度-活力曲线得IC50[2]
- Ras法尼基化Western blot：
1. SK-MES-1细胞（2×105细胞/孔，6孔板）用替匹法尼（Tipifarnib） （10~200 nM）处理48小时[2]
2. 含蛋白酶抑制剂的RIPA缓冲液裂解细胞，4°C、12,000×g离心15分钟，BCA法测蛋白浓度[2]
3. 每泳道30 μg蛋白经12% SDS-PAGE分离后转移至PVDF膜，加入抗法尼基化H-Ras一抗（1:1000）和抗总H-Ras一抗（1:1000，内参）孵育；HRP标记二抗（1:5000）结合后ECL显影，ImageJ定量条带强度[2]
- 外泌体分泌实验：
1. MDA-MB-231细胞（1×106细胞/皿，10 cm培养皿）在无血清培养基中用替匹法尼（Tipifarnib） （1 μM、10 μM）处理48小时[3]
2. 收集细胞上清，300×g离心10分钟（去除细胞）、10,000×g离心30分钟（去除细胞碎片），随后4°C、100,000×g超速离心70分钟分离外泌体[3]
3. 外泌体沉淀用PBS重悬，流式细胞术（抗CD63-PE抗体染色）和Western blot（抗TSG101抗体）检测外泌体量，相对溶剂组计算分泌抑制率[3]

Animal/Disease Models: GalN/LPS challenge mouse[5]
Doses: 10 mg/kg; while chanllenge with GalN (400 mg/kg; IP) and LPS (32 g/kg)
Route of Administration: IP; 60 min before challenge
Experimental Results: Protected primary hepatocytes from GalN/tumor necrosis factor-induced cell death. Inhibited caspase 3 activation and upregulating antiapoptotic proteins.

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Nude mouse xenograft model :
1. Animals and grouping: Female BALB/c nude mice (6–8 weeks old, 18–22 g) were randomly divided into 3 groups (n=6 per group): vehicle control (0.5% CMC-Na), Tipifarnib 25 mg/kg bid, Tipifarnib 50 mg/kg bid [2]
2. Tumor induction: 1×106 SK-MES-1 cells (suspended in 100 μL PBS:Matrigel = 1:1) were subcutaneously injected into the right flank of each mouse. Experiments started when tumors reached ~100 mm³ (day 0) [2]
3. Drug preparation and administration: Tipifarnib was dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) to prepare suspensions. Mice received oral gavage twice daily (8:00 and 20:00) for 14 days, with a dosing volume of 10 mL/kg [2]
4. Sample collection and detection:
- Tumor volume: Measured every 3 days using calipers, calculated as V = (length × width²) / 2.
- Body weight: Recorded every 3 days to monitor toxicity.
- Tumor weight: On day 14, mice were euthanized, tumors were dissected and weighed. Tumor growth inhibition (TGI) rate = [1 – (tumor weight of treated group / tumor weight of vehicle group)] × 100%.
- Tumor tissue analysis: A portion of tumor tissue was lysed for Western blot detection of farnesylated H-Ras [2]

Oral absorption:
- In rats, oral administration of Tipifarnib (10 mg/kg) showed an oral bioavailability (F) of 35%, with a time to reach maximum concentration (Tmax) of 1.5 hours and maximum plasma concentration (Cmax) of 89 ng/mL [1]
- Distribution:
- In nude mice bearing SK-MES-1 xenografts, 2 hours after oral administration of 50 mg/kg Tipifarnib, tumor tissue concentration was 2.3-fold higher than plasma concentration (tumor: 156 ng/g; plasma: 68 ng/mL) [2]
- Elimination:
- In rats, the elimination half-life (t1/2) of Tipifarnib was 4.2 hours (intravenous dose: 5 mg/kg). Fecal excretion accounted for 65% of the dose, and urinary excretion accounted for 12% within 72 hours [1]

In vitro cytotoxicity selectivity:
- In normal human lung fibroblasts (MRC-5), Tipifarnib showed low cytotoxicity: IC50 = 500 nM (72-hour MTT), which was ~28-fold higher than that in SK-MES-1 cancer cells (18 nM), indicating selective antitumor activity [2]
- In vivo safety:
- In nude mice treated with Tipifarnib (up to 50 mg/kg bid for 14 days):
- No significant changes in serum liver function markers (ALT, AST) or kidney function markers (BUN, creatinine);
- Peripheral blood leukocyte count remained within the normal range (no myelosuppression);
- Body weight loss was <5% (no severe toxicity) [2]
- Plasma protein binding:
- Tipifarnib showed high plasma protein binding (>97%) in human, rat, and mouse plasma, determined by equilibrium dialysis (37°C, pH 7.4) [1]

[1]. Inhibition of farnesyltransferase: a rational approach to treat cancer? J Enzyme Inhib Med Chem. 2007 Apr;22(2):127-40.

[2]. Characterization of the antitumor effects of the selective farnesyl protein transferase inhibitor R115777 in vivo and in vitro. Cancer Res. 2001 Jan 1;61(1):131-7.

[3]. High-throughput screening identified selective inhibitors of exosome biogenesis and secretion: A drug repurposing strategy for advanced cancer. Sci Rep. 2018 May 25;8(1):8161.

Tipifarnib is a quinolone that is 1-methylquinolin-2-one which carries a 3-chlorophenyl and an amino(4-chlorophenyl)(1-methyl-imidazol-5-yl)methyl groups at the 4 and 6 positions, respectively (the R-isomer). It has a role as an antineoplastic agent, an EC 2.5.1.58 (protein farnesyltransferase) inhibitor and an apoptosis inducer. It is a quinolone, a member of monochlorobenzenes, a member of imidazoles and a primary amino compound.
Tipifarnib (R-115777) is a substance that is being studied in the treatment of acute myeloid leukemia (AML) and other types of cancer. It belongs to the family of drugs called farnesyltransferase inhibitors. It is also called Zarnestra. In June 2005, the FDA issued a Not Approvable Letter for Zarnestra.
Tipifarnib is a nonpeptidomimetic quinolinone with potential antineoplastic activity. Tipifarnib binds to and inhibits the enzyme farnesyl protein transferase, an enzyme involved in protein processing (farnesylation) for signal transduction. By inhibiting the farnesylation of proteins, this agent prevents the activation of Ras oncogenes, inhibits cell growth, induces apoptosis, and inhibits angiogenesis. (NCI04)

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Drug Indication
Investigated for use/treatment in colorectal cancer, leukemia (myeloid), pancreatic cancer, and solid tumors.
Treatment of head and neck epithelial malignant neoplasms

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Mechanism of Action
The farnesyltransferase inhibitors (FTIs) are a class of experimental cancer drugs that target protein farnesyltransferase with the downstream effect of preventing the proper functioning of the Ras protein, which is commonly abnormally active in cancer. After translation, RAS goes through four steps of modification: isoprenylation, proteolysis, methylation and palmitoylation. Isoprenylation involves the enzyme farnesyltransferase (FTase) transferring a farnesyl group from farnesyl pyrophosphate (FPP) to the pre-RAS protein. Also, a related enzyme geranylgeranyltransferase I (GGTase I) has the ability to transfer a geranylgeranyl group to K and N-RAS. Farnesyl is necessary to attach RAS to the cell membrane. Without attachment to the cell membrane, RAS is not able to transfer signals from membrane receptors (Reuter et al., 2000).

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Pharmacodynamics
R115777, a nonpeptidomimetic farnesyl transferase inhibitor, suppresses the growth of human pancreatic adenocarcinoma cell lines. This growth inhibition is associated with modulation in the phosphorylation levels of signal transducers and activators of transcription 3 (STAT3) and extracellular signal-regulated kinases (ERK).

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Tipifarnib (R115777) is a synthetic, oral, and selective farnesyl protein transferase (FTase) inhibitor, developed as the first-in-class FTase inhibitor for the treatment of Ras-mutated cancers [1][2]
- Its core mechanism: FTase catalyzes the farnesylation of Ras proteins (H-Ras, K-Ras, N-Ras), a post-translational modification required for Ras localization to the cell membrane and activation of oncogenic signaling (e.g., MAPK/ERK pathway). Tipifarnib inhibits FTase, blocking Ras farnesylation and membrane localization, thereby suppressing Ras-mediated cancer cell proliferation and survival [1][2]
- The selective inhibition of exosome biogenesis by Tipifarnib (via Rab27a farnesylation inhibition) suggests an additional anti-metastatic effect, as exosomes promote cancer cell invasion, angiogenesis, and immune evasion. This expands its potential therapeutic applications beyond Ras-mutated cancers [3]
- Preclinical studies confirm that Tipifarnib exhibits potent antitumor activity in H-Ras-mutated tumors, with favorable oral bioavailability and manageable toxicity, supporting its clinical development for Ras-driven malignancies [1][2]

C27H22CL2N4O

489.4

488.117

192185-72-1

Tipifarnib (S enantiomer);192185-71-0

159324

White to light yellow solid powder

1.3±0.1 g/cm3

681.7±55.0 °C at 760 mmHg

211-213ºC (dec.)

366.1±31.5 °C

0.0±2.1 mmHg at 25°C

1.672

4.94

65.84

1

3

4

34

785

1

CN1C=NC=C1[C@@](C2=CC=C(C=C2)Cl)(C3=CC4=C(C=C3)N(C(=O)C=C4C5=CC(=CC=C5)Cl)C)N

PLHJCIYEEKOWNM-HHHXNRCGSA-N

InChI=1S/C27H22Cl2N4O/c1-32-16-31-15-25(32)27(30,18-6-9-20(28)10-7-18)19-8-11-24-23(13-19)22(14-26(34)33(24)2)17-4-3-5-21(29)12-17/h3-16H,30H2,1-2H3/t27-/m1/s1

6-[(R)-amino-(4-chlorophenyl)-(3-methylimidazol-4-yl)methyl]-4-(3-chlorophenyl)-1-methylquinolin-2-one

R115777; R 115777; R-115777; LX81; NSC702818; D03720; trade name Zarnestra

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)

配方 1 中的溶解度: ≥ 1.43 mg/mL (2.92 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 14.3 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: 1.43 mg/mL (2.92 mM) 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), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 14.3mg/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: ≥ 1.43 mg/mL (2.92 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 14.3 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 4 中的溶解度: 15% Captisol+citrate vehicle: 5 mg/mL

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配方 5 中的溶解度: 10 mg/mL (20.43 mM) in 20% HP-β-CD/10 mM citrate pH 2.0 (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。