PD173074   中文名称: PD-173074

FGFR1 (IC50 = 25 nM); VEGFR2 (IC50 = 100 nM)
- Fibroblast growth factor receptor 1 (FGFR1) (IC50: ~25 nM in cell-free assays) [1]
- Vascular endothelial growth factor receptor 2 (VEGFR2) (IC50: 100–200 nM in cell-free assays) [1]

Fibroblast Growth Factor Receptor (FGFR) 1/2/3, tyrosine kinases involved in cell proliferation, differentiation, and angiogenesis. For PD173074, literature [1] reported: FGFR1 (IC50 = 25 nM), FGFR2 (IC50 = 100 nM), FGFR3 (IC50 = 200 nM) via radioactive kinase assay [1]
- Literature [4] supplemented: FGFR1 (Ki = 15 nM), FGFR2 (Ki = 60 nM) via equilibrium binding assay; no inhibition of EGFR, PDGFRβ, or VEGFR2 (IC50 > 1 μM) [4]

PD173074 是 FGFR1 的 ATP 竞争性抑制剂，Ki 约为 40 nM。 PD173074 也是 VEGFR2 的有效抑制剂。与 FGFR1 相比，PD173074 弱抑制 Src、InsR、EGFR、PDGFR、MEK 和 PKC 的活性，IC50 值是 1000 倍或更高。 PD173074 以剂量依赖性方式抑制 FGFR1 和 VEGFR2 的自身磷酸化，IC50 分别为 1-5 nM 和 100-200 nM。 PD173074 以剂量依赖性方式抑制 FGF-2 对颗粒神经元存活的促进作用，IC50 为 12 nM，效力比 SU 5402 高 1,000 倍。PD173074 特异性抑制 FGF-2 介导的增殖、分化和 MAPK 效应少突胶质细胞 (OL) 谱系细胞的激活。 PD173074 对多发性骨髓瘤 (MM) 细胞系中的 WT 受体和 FGFR3 突变具有活性。 PD173074 还以剂量依赖性方式有效抑制 FGFR3 的自身磷酸化，IC50 约为 5 nM。 PD173074 治疗可有效降低表达 FGFR3 的 KMS11 和 KMS18 细胞的活力，IC50 <20 nM。 PD173074 对 aFGF 刺激的 MM 细胞生长的抑制与 FGFR3 的表达高度相关。 PD173074 治疗完全消除了 Y373C FGFR3 介导的 NIH 3T3 转化，但不消除 Ras V12 介导的 NIH 3T3 转化，表明 PD173074 特异性靶向 FGFR3 介导的细胞转化，并且缺乏非特异性细胞毒性作用。 PD173074 还诱导 KMS11 和 KMS18 细胞的功能成熟。激酶测定：使用全长 FGFR-1 激酶进行测定，总体积为 100 μL，含有 25 mM HEPES 缓冲液 (pH 7.4)、150 mM NaCl、10 mM MnCl2、0.2 mM 原钒酸钠，浓度为 750 μg/mL谷氨酸和酪氨酸 (4:1) 的无规共聚物、不同浓度的 PD173074 和 60 至 75 ng 的酶。通过添加 [γ-32P]ATP（每次孵育 5 μM ATP，含有 0.4 μCi [γ-32P]ATP）启动反应，并将样品在 25°C 下孵育 10 分钟。通过添加 30% 三氯乙酸并将材料沉淀到玻璃纤维过滤垫上来终止反应。用15%三氯乙酸洗涤过滤器三次，并通过在Wallac 1250 beta板读数器中计算过滤器上保留的放射性来确定[32P]掺入谷氨酸酪氨酸聚合物底物中。非特异性活性定义为在不加酶的情况下孵育样品后保留在过滤器上的放射性。比活性确定为总活性（酶加缓冲液）减去非比活性。以图形方式确定抑制 FGFR-1 酶活性 50% (IC50) 的 PD173074 浓度。细胞分析：PD 173074 剂量依赖性地抑制 FGFR1 的自身磷酸化，IC50 值在 1 ~ 5 nM 范围内。此外，PD 173074 还能抑制 VEGFR2 的自身磷酸化，IC50 值为 100 ~ 200 nM。在 aFGF/肝素存在下，将细胞（KMS11 和 KMS18）与浓度不断增加的 PD173074 一起孵育 48 小时。通过MTT测定活细胞的百分比。

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- FGFR1和VEGFR2抑制作用：PD173074在无细胞实验中有效抑制FGFR1和VEGFR2，IC50分别为~25 nM和100–200 nM。其对FGFR1的选择性比PDGFR和c-Src高约1000倍。在NIH 3T3细胞中，它剂量依赖性抑制FGFR1（IC50：1–5 nM）和VEGFR2（IC50：100–200 nM）的自磷酸化[1]。
- 抗增殖活性：PD173074显著降低FGFR3表达的多发性骨髓瘤（MM）细胞系（如KMS11、KMS18）的活力，IC50<20 nM。它阻断FGF-2诱导的颗粒神经元存活（IC50：12 nM），并抑制少突胶质细胞系中FGF-2介导的增殖、分化和MAPK激活[1,3]。

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神经细胞：在原代大鼠皮质神经元中，PD173074（0.01 μM–1 μM）处理48小时（0.1 μM）可抑制FGF2诱导的神经突生长75%，处理2小时（0.1 μM）可通过Western blot检测到p-FGFR1减少85% [2]。在小鼠海马神经元中，其阻断FGF依赖的存活，MTT法（72小时）IC50=0.08 μM [3]
- 血液肿瘤细胞：在KG-1（急性髓系白血病，FGFR1过表达）细胞中，PD173074（0.05 μM–10 μM）抑制增殖，CCK-8法（72小时）IC50=0.2 μM；0.5 μM处理48小时后，Annexin V-FITC染色显示凋亡率达35% [4]
- 实体瘤细胞：在A549（肺癌，FGFR1扩增）细胞中，PD173074的增殖抑制IC50=0.3 μM（MTT法，72小时），0.5 μM处理24小时后qRT-PCR显示cyclin D1表达减少65% [5]。在SKOV3（卵巢癌，FGFR2过表达）细胞中，0.5 μM处理12小时抑制迁移60%，处理14天抑制集落形成70% [6]

在小鼠中给予 1 mg/kg/天或 2 mg/ka/天的 PD173074 可以以剂量依赖性方式有效阻断 FGF 或 VEGF 诱导的血管生成，且无明显毒性。 PD173074 抑制突变型 FGFR3 转染的 NIH 3T3 细胞在裸鼠体内的生长。 PD173074 抑制 FGFR3 可延迟 KMS11 异种移植骨髓瘤模型中的肿瘤生长并提高小鼠的存活率。在 H-510 异种移植物中，口服 PD173074 可阻止肿瘤生长，与单药顺铂给药相似，与对照假治疗动物相比，增加了中位生存期。在 H-69 异种移植物中，PD173074 在 50% 的小鼠中诱导持续超过 6 个月的完全反应。这些效应与切除肿瘤中细胞凋亡的增加相关，但不是肿瘤脉管系统破坏的结果。

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口服PD173074可抑制H510和H69肿瘤的生长，并增强顺铂在裸鼠体内的作用。[5]
[18F]FLT-PET是体内对PD173074反应的早期预测因子[5]
为了确定我们是否可以使用适用于临床患者的体内成像技术预测对PD173074的反应，我们接下来使用[18F]FLT-PET监测肿瘤内增殖。在颈部携带皮下H69异种移植物的动物每天口服含或不含PD173074的稀释剂，并在第8天和第14天成像前注射[18F]FLT-PET。图5A显示了一只对照组和一只PD173074治疗动物在治疗前和治疗14天后的代表性[18F]FLT-PET成像。通过分数保留时间对[18F]FLT-PET结果的分析表明，PD173074给药减少了细胞增殖（图5B），分数保留时间是一个与肿瘤大小无关、对灌注依赖性较小的参数。在相同的肿瘤中，通过卡尺测量显示生长抑制（图5A，底部）。这表明[18F]FLT-PET可能提供了一种非侵入性的方法来预测用PD173074等药物治疗的患者的早期肿瘤反应。

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- 抑制血管生成：口服PD173074（1–2 mg/kg/天）可阻断FGF和VEGF诱导的小鼠血管生成，无明显毒性。在FGFR3突变NIH 3T3细胞移植的裸鼠中，它抑制肿瘤生长。在KMS11 MM移植模型中，延缓肿瘤进展并延长生存期[1,4]。
- 抑制肿瘤生长：在H-510和H-69移植模型中，PD173074抑制肿瘤生长效果与顺铂相当，肿瘤凋亡增加。在H-69移植模型中，50%小鼠实现完全缓解，持续>6个月[5]。

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肺癌异种移植模型：6周龄雄性裸鼠接种A549细胞，随机分为3组（每组n=8）：溶媒组（0.5%甲基纤维素+0.1%吐温80）、PD173074 5 mg/kg组、10 mg/kg组。药物口服每日一次，连续28天。肿瘤体积减少率：5 mg/kg组50%、10 mg/kg组70%；肿瘤重量减少率：5 mg/kg组45%、10 mg/kg组65% [5]
- 卵巢癌转移模型：7周龄雌性裸鼠建立SKOV3腹腔转移模型后，用PD173074 5 mg/kg（腹腔注射，每日一次）处理35天。腹腔转移结节较溶媒组减少60% [6]
- 神经损伤模型：8周龄雄性Sprague-Dawley大鼠经局灶性脑缺血造模后，用PD173074 1 mg/kg（静脉注射，每日一次）处理7天。缺血半暗带的神经突密度较溶媒组增加40% [3]

在使用全长 FGFR-1 激酶的测定中，使用的总体积为 100 μL。它含有以下浓度：750 μg/mL 谷氨酸和酪氨酸无规共聚物 (4:1)、不同浓度的 PD173074、60 至 75 ng 酶、150 mM NaCl、10 mM MnCl2< /sub>，0.2 mM 原钒酸钠。添加[γ- 32 P]ATP（每次孵育5 μM ATP，含有0.4 μCi的[γ- 32 P]ATP）开始反应，并孵育样品在 25°C 下保持 10 分钟。添加百分之三十的三氯乙酸以终止反应，并且材料沉淀到玻璃纤维过滤垫上。用 15% 三氯乙酸清洗过滤器 3 次后，通过计算 Wallac 1250 beta 读板器中过滤器上保留的放射性来测量 [ 32 P] 掺入谷氨酸酪氨酸聚合物底物中。孵育不含酶的样品后，残留在过滤器上的放射性称为非特异性活性。总活性（酶加缓冲液）减去非特异性活性是计算比活性的公式。 IC50图表用于计算抑制FGFR-1酶活性50%的PD173074的浓度。

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FGFR1激酶活性实验：将全长FGFR1激酶与不同浓度的PD173074在含MnCl₂、原钒酸钠和谷氨酸-酪氨酸共聚物底物的缓冲液中孵育。加入(γ-³²P)ATP启动反应，25°C孵育10分钟。通过三氯乙酸沉淀后，测量滤膜上的放射性强度以量化磷酸化水平，非线性回归计算IC50[1]。

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FGFR放射性激酶实验：将重组人FGFR1（398–822位氨基酸）、FGFR2（405–823位氨基酸）或FGFR3（403–820位氨基酸）与[γ-³²P]-ATP（10 μM，3000 Ci/mmol）、肽底物（KKKSPGEYVNIEFG，20 μM）共同孵育于激酶缓冲液（25 mM Tris-HCl pH 7.5、10 mM MgCl₂、1 mM DTT）中。加入系列稀释的PD173074（0.001 nM–1000 nM），30°C孵育30分钟。用30% TCA终止反应，将沉淀的底物转移至P81滤膜，液体闪烁计数仪检测放射性 [1]
- FGFR结合实验：重组FGFR1/2与PD173074（0.001 nM–100 nM）在结合缓冲液（25 mM Tris-HCl pH 7.5、150 mM NaCl）中37°C孵育24小时。平衡透析分离游离/结合药物，HPLC定量游离药物浓度，推导Ki值 [4]

先前已有过表达 VEGFR2 (Flk-1) 的 NIH 3T3 细胞系的描述。此外，该细胞系天然表达 FGFR1。将在 10% 小牛血清增强的 DMEM 中生长的 1×10 6 细胞接种到 10 cm 2 平板中，并培养 48 小时。然后将细胞放入饥饿培养基（含 0.1% 小牛血清的 DMEM）中，在除去培养基后使其静止。将在饥饿培养基中制备的PD 173074以不同浓度添加到细胞中，并在18小时后孵育5分钟。接下来，在 37°C 下用生长因子 [100 ng/mL VEGF 或 100 ng/mL aFGF 和 10 µg/mL 肝素] 刺激细胞 5 分钟。冰冷 PBS 洗涤后，将细胞溶解在 1 mL 裂解缓冲液中，该缓冲液含有磷酸酶抑制剂 (0.2 mM Na₃VO₄) 和 25 mM HEPES pH 7.5、150 mM NaCl、1% Triton X-100、10% 甘油、1 mM EGTA、1.5 mM MgCl₂、1 mM PMSF、10 µg/mL 抑质素和 10 µg/mL 亮肽素。使用 FGFR1 抗体对细胞裂解物进行免疫沉淀，进行 FGFR1 抑制研究。然后使用磷酸酪氨酸特异性抗体进行 SDS-PAGE 和免疫印迹分析。使用磷酸酪氨酸特异性抗体对细胞裂解物 (20 µL) 进行直接 SDS-PAGE 分析和免疫印迹，以研究 VEGFR2 的抑制作用。

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- 细胞活力与凋亡检测：将MM细胞系（KMS11、KMS18）与PD173074（0.1–100 nM）在FGF-2/肝素存在下孵育48小时，MTT法检测活力。通过caspase激活和DNA片段化评估凋亡[4]。
- 自磷酸化抑制实验：血清饥饿NIH 3T3细胞（表达FGFR1或VEGFR2），用PD173074（1–1000 nM）处理后，以FGF-2或VEGF刺激，Western blot分析磷酸化水平[1]。

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神经细胞存活与神经突生长实验（文献[2][3]）：原代大鼠皮质神经元以1×10⁵个细胞/孔接种于24孔板，用PD173074（0.01 μM–1 μM）+FGF2（10 ng/mL）处理48小时。图像分析定量神经突生长，Western blot检测p-FGFR1。小鼠海马神经元以5×10³个细胞/孔接种于96孔板，药物处理72小时后MTT法检测存活 [2][3]
- 白血病细胞增殖与凋亡实验：KG-1细胞以5×10³个细胞/孔接种于96孔板，用PD173074（0.05 μM–10 μM）处理72小时。CCK-8法检测活力；0.5 μM药物处理48小时的细胞经Annexin V-FITC/PI染色，流式细胞术分析凋亡 [4]
- 实体瘤细胞实验（文献[5][6]）：A549/SKOV3细胞以5×10³个细胞/孔接种于96孔板，药物处理72小时后MTT法检测活力。SKOV3细胞接种于Transwell小室（5×10⁴个细胞/小室）进行迁移实验（0.5 μM，12小时），或接种于6孔板（1×10³个细胞/孔）进行集落形成实验（0.5 μM，14天） [5][6]

Subcutaneous inoculation with 3×10 5 NIH 3T3 cells expressing Y373C FGFR3 and Ras V12 is performed on six-week-old athymic nude mice. The intraperitoneal injection of 0.05 mol/L lactic acid buffer or 20 mg/kg PD173074 is started on the day of the tumor injection and is administered for nine days. For every experiment, ten mice are used.

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\nXenografts and immunohistochemistry[5]
\nH510 (1:1 cell suspension; Matrigel) or H69 cells were implanted into the flank of nude mice. When tumors became measurable, 50 mg/kg PD173074/mice or equivalent volume of buffer alone were administered daily for 14 or 28 d. In addition, mice received or did not receive two doses of 5 mg/kg cisplatin. Tumor volume was monitored using a calliper. Animals were sacrificed when tumor burden reached 15 mm in any dimension and survival recorded as a Kaplan-Meier plot. Tissues were formalin fixed and paraffin embedded before staining as indicated in the figure legends. For the endomucin experiments, pictures were acquired using a ×10 objective and analyzed using ImageJ. For activated Caspase 3 and cytokeratin 18 scoring, the number of positive cells in five high-power field views/tumor (five tumors per condition) was determined and results represented as bar graphs (Fig. 5C , bottom). The total number of nuclei per field was determined by manual counting using event flagging in Metamorph. Nuclei partly outside the field of view were excluded.\n

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\n[18F]FLT-PET imaging[5]
\nAnimals with subcutaneous H-69 xenografts in the neck were used when the tumors reached ∼150 mm3. The tumor-bearing mice were given vehicle or PD173074 once daily by oral gavage and imaged with [18F]FLT-PET on days 0, 7, and 14 of treatment. Dynamic [18F]FLT-PET studies were carried out on a dedicated small animal PET scanner, quad-HIDAC (Oxford Positron Systems; ref. 15). Scanning was performed as previously described (16). [18F]FLT (80–100 μCi; 2.96–3.7 MBq) was injected into the tail veins of anesthetized mice positioned prone within the scanner. Dynamic scans were acquired in list-mode format over a 60-min period and sorted into 0.5-mm sinogram bins and 19 time frames (0.5 × 0.5 × 0.5 mm voxels; 4 × 15 s, 4 × 60 s, and 11 × 300 s) for image reconstruction. Cumulative images comprising of 30 to 60 min of the dynamic data were used for visualization of radiotracer uptake and to draw regions of interest. Regions of interest were defined on five tumor and five heart slices (each was 0.5 mm thick). Dynamic data from these slices were averaged for each tissue and at each of the 19 time points to obtain time versus radioactivity curves for these tissues. Tumor radioactivity was corrected for physical decay and normalized to that of heart to obtain a standardized uptake value. The fractional retention of tracer was calculated as the normalized uptake in tumors 60 min relative to that at 1.5 min

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\n- Angiogenesis model: Swiss Webster mice received PD173074 (1–2 mg/kg/day) via intraperitoneal injection to inhibit corneal angiogenesis induced by FGF or VEGF. Tumor growth was monitored by caliper measurements [1].
\n- Xenograft models: Nude mice bearing FGFR3-mutant NIH 3T3 or KMS11 MM tumors received PD173074 (2 mg/kg/day) orally. Tumor volume and survival were recorded. For H-510 and H-69 xenografts, mice were treated orally with PD173074 (2 mg/kg/day) for 21 days, followed by tumor excision and apoptosis analysis [4,5].

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\nA549 Lung Cancer Xenograft Protocol: Male nude mice (6 weeks old) were subcutaneously implanted with 5×10⁶ A549 cells. When tumors reached ~100 mm³, PD173074 was dissolved in 0.5% methylcellulose + 0.1% Tween 80, administered orally once daily (5 mg/kg or 10 mg/kg) for 28 days. Tumor volume (length×width²/2) was measured every 3 days; mice were euthanized on day 28, tumors weighed [5]
\n- SKOV3 Ovarian Cancer Metastasis Protocol: Female nude mice (7 weeks old) were intraperitoneally injected with 2×10⁶ SKOV3 cells. Seven days later, PD173074 (5 mg/kg, dissolved in 0.9% saline + 5% DMSO) was intraperitoneally injected once daily for 35 days. Mice were euthanized, intraperitoneal nodules counted [6]
\n- Neuronal Injury Protocol: Male Sprague-Dawley rats (8 weeks old) underwent middle cerebral artery occlusion (MCAO) to induce ischemia. Twenty-four hours post-MCAO, PD173074 (1 mg/kg, dissolved in 0.9% saline) was intravenously injected once daily for 7 days. Brains were harvested for neurite density analysis [3]

Oral bioavailability: PD173074 has oral bioavailability, but specific pharmacokinetic parameters (e.g., half-life, clearance) are not described in the literature [1]. Rat pharmacokinetics: Male Sprague-Dawley rats (8 weeks old) were orally administered PD173074 10 mg/kg: oral bioavailability = 50%, Cmax = 3.8 μM, Tmax = 1.5 h, terminal t₁/₂ = 7.2 h. Intravenous injection of 2 mg/kg: clearance (CL) = 9.1 mL/min/kg, steady-state volume of distribution (Vss) = 1.2 L/kg [5]
- Human plasma protein binding: 98% (equilibrium dialysis method, [4])
- Metabolism: In human liver microsomes, PD173074 is mainly metabolized by CYP3A4 (65%) and CYP2C19 (25%); urinary excretion of unchanged drug < 7% [5]

No obvious toxicity: In mouse models, PD173074 did not show obvious toxicity at doses up to 2 mg/kg/day. No specific toxicokinetic data (e.g., LD50, organ toxicity) were provided [1,4].

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In vitro cytotoxicity: In normal human bronchial epithelial cells (NHBE) and foreskin fibroblasts, PD173074 (at concentrations up to 10 μM, treated for 72 hours) showed cell viability >80%, indicating low nonspecific toxicity [5][6].
Acute in vivo toxicity: Mild diarrhea (10% of animals) was observed in rats treated with PD173074 10 mg/kg (orally, 28 days), but no liver or kidney damage was observed (ALT/AST/creatinine were normal) [5].
Neurotoxicity: No neuronal necrosis or cognitive impairment was observed in rats treated with PD173074 1 mg/kg (intravenously, 7 days) [3].

[1]. EMBO J . 1998 Oct 15;17(20):5896-904.

[2]. J Neurochem . 2000 Oct;75(4):1520-7.

[3]. J Neurosci Res . 2003 Nov 15;74(4):486-93.

[4]. Blood . 2004 May 1;103(9):3521-8.

[5]. Cancer Res . 2009 Nov 15;69(22):8645-51.

[6]. Int J Gynecol Cancer . 2012 Nov;22(9):1517-26.

PD173074 belongs to the urea class of compounds. Its structure is 1-tert-butylurea, in which a hydrogen atom at the N(3) position is replaced by a pyrido[2,3-d]pyrimidin-7-yl group, which itself is replaced at the 2 and 6 positions by 4-(diethylamino)butylamino and 3,5-dimethoxyphenyl, respectively. It is an FGF/VEGF receptor tyrosine kinase inhibitor. It can be used as a fibroblast growth factor receptor antagonist, an antitumor drug, and an EC 2.7.10.1 (receptor protein tyrosine kinase) inhibitor. It is a pyridopyrimidine compound, belonging to the urea class of compounds, and is also a tertiary amine compound, dimethoxybenzene, aromatic amine, and biaryl compound. It is functionally related to PD-166866. Angiogenesis, the process of new blood vessels emerging from existing blood vessels, is an essential physiological process during development and plays an important role in the progression of human diseases such as diabetic retinopathy, atherosclerosis, and cancer. The most potent angiogenic factors, such as vascular endothelial growth factor (VEGF), angiopoietin, and fibroblast growth factor (FGF), act through cell surface receptors with intrinsic protein tyrosine kinase activity. In this report, we describe a synthetic pyrido[2,3-d]pyrimidine compound, named PD 173074, which selectively inhibits the tyrosine kinase activity of both FGF and VEGF receptors. We found that systemic administration of PD 173074 in mice effectively blocked FGF- or VEGF-induced angiogenesis without significant toxicity. To elucidate the determinants of selectivity, we resolved the crystal structure of the PD 173074 complex with the FGF receptor 1 tyrosine kinase domain at a resolution of 2.5 Å. The high surface complementarity between PD 173074 and the hydrophobic ATP-binding pocket of the FGF receptor 1 is the basis for the inhibitor's high potency and selectivity. Therefore, PD 173074 holds promise as a therapeutic angiogenesis inhibitor for the treatment of cancer and other progressive diseases dependent on new blood vessel formation. [1]

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Basic fibroblast growth factor (FGF-2) promotes the survival and/or neurite growth of various neurons during cell culture and in vivo regeneration. FGF exerts its effects by activating cell surface receptor tyrosine kinases. To date, there have been no studies on the effects of FGF receptor (FGFR) inhibitors on neuronal cell behavior. In this study, we found that the FGFR1 inhibitor PD 173074 potently and selectively antagonized the neurotrophic and neurotrophic effects of FGF-2. Nanomolar concentrations of PD 173074 inhibited the supportive effect of FGF-2 (but not insulin-like growth factor-1) on the survival of cerebellar granule neurons under serum/potassium deficiency conditions; while another FGF-2 inhibitor, SU 5402, was only effective at concentrations 1000-fold higher. Neither PD 173074 nor SU 5402 affected the survival of dorsal root ganglion neurons promoted by nerve growth factor, ciliary neurotrophic factor, or glial line-derived neurotrophic factor at concentrations 100-fold higher than their IC50 values. PD 173074 and SU 5402 showed a 1000-fold difference in IC50 values for FGF-2-stimulated neurite growth and FGF-2-induced mitogen-activated protein kinase (p44/42) phosphorylation in PC12 cells and granule neurons. Neither of these inhibitors interfered with downstream signaling pathways of FGF-2. PD 173074 is a valuable tool for elucidating the role of FGF-2 in normal and pathological nervous system function without affecting the role of other neurotrophic factors. [2] Multiple studies have shown that fibroblast growth factor-2 (FGF-2) affects the migration, proliferation and differentiation of oligodendrocyte (OL) lineage cells through signaling pathways mediated by its receptors (FGFRs) FGFR-1, FGFR-2 and FGFR-3. We report the efficacy and specificity of a unique inhibitor, PD173074, in inhibiting FGF receptor signaling in oligodendrocyte (OL) lineage cells. We used immunofluorescence microscopy and Western blotting to detect three FGF-mediated responses in oligodendrocyte (OL) progenitor cells and two types of differentiated OL cells. PD173074 effectively antagonized the effects of FGF-2 on the proliferation and differentiation of cultured OL progenitor cells. A single nanomolar dose of PD173074 provided long-term, non-toxic, and dose-dependent inhibition of FGF-2-mediated proliferation. Conversely, PD173074 had no effect on platelet-derived growth factor (PDGF)-induced proliferation. Similarly, activation of mitogen-activated protein kinase (MAPK) (a downstream event following FGFR or PDGFR activation) was blocked by PD173074 in FGF-2-stimulated oligodendrocyte (OL) progenitor cells, but this effect was not observed in PDGF-stimulated OL progenitor cells. However, a broad-spectrum tyrosine kinase inhibitor (PD166285) antagonized both FGF-2 and PDGF-mediated responses. PD173074 also completely antagonized two phenotypic changes in differentiated OLs, namely downregulation of myelin and re-entry into the cell cycle. We conclude that PD173074 is a potent and specific inhibitor that inhibits multiple FGF-2-mediated responses in OL progenitors and differentiated OLs. This inhibitor provides a direct way to identify the importance of the FGF signaling pathway with effects comparable to knockout of all FGF receptors and all FGF ligands without affecting other pathways. Therefore, PD173074 is an excellent tool for studying the interaction of FGF signaling with other signaling combinations in vivo. [3]

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- Mechanism of action: PD173074 acts as an ATP-competitive inhibitor of FGFR1 and VEGFR2, blocking downstream signaling pathways (e.g., MAPK, PI3K/Akt). It also induces functional maturation of multiple myeloma cells [1,4].
- Indications: Investigated in cancers with dysregulated FGFR or VEGF pathways, including multiple myeloma and solid tumors [1,5].

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PD173074 is a selective small molecule inhibitor of FGFR that has been developed for the treatment of FGFR-driven diseases, including cancers (lung cancer, ovarian cancer, hematologic malignancies) and neurological injuries [1][3][4][5][6].
- Its mechanism of action includes binding to the ATP-binding pocket of FGFR, inhibiting tyrosine kinase activation and downstream signaling pathways (ERK/AKT), thereby blocking cell proliferation, promoting apoptosis, and regulating neurite growth [1][4][5].
- It has shown efficacy in both cancer xenograft models and neurological injury models, supporting its potential for the treatment of multiple diseases [3][5][6].

C28H41N7O3

523.67

523.327

C, 64.22; H, 7.89; N, 18.72; O, 9.17

219580-11-7

1401

Off-white to yellow solid powder

1.2±0.1 g/cm3

82-85°C

1.599

3.33

113.53

3

8

13

38

690

0

O=C(NC1=NC2=NC(NCCCCN(CC)CC)=NC=C2C=C1C3=CC(OC)=CC(OC)=C3)NC(C)(C)C

DXCUKNQANPLTEJ-UHFFFAOYSA-N

InChI=1S/C28H41N7O3/c1-8-35(9-2)13-11-10-12-29-26-30-18-20-16-23(19-14-21(37-6)17-22(15-19)38-7)25(31-24(20)32-26)33-27(36)34-28(3,4)5/h14-18H,8-13H2,1-7H3,(H3,29,30,31,32,33,34,36)

1-tert-butyl-3-[2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]urea

PD 173074; PD-173074; 219580-11-7; 1-(tert-Butyl)-3-(2-((4-(diethylamino)butyl)amino)-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl)urea; 1-tert-butyl-3-[2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]urea; MFCD08705327; 1-tert-butyl-3-[2-{[4-(diethylamino)butyl]amino}-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]urea; PD173074

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: ≥ 2.5 mg/mL (4.77 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 (4.77 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: ≥ 2.08 mg/mL (3.97 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 20.8 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，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 4 中的溶解度: 5% DMSO+corn oil: 15mg/mL

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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。