125 I-CXCL12-CXCR4 ( IC50 = 44 nM ); 125 I-CXCL12-CXCR7; HIV-1 ( EC50 = 1-10 nM ); HIV-2 ( IC50 = 1-10 nM )
CXCR4 receptor (Ki = 4.1 nM, human; IC50 = 7.5 nM for CXCL12 binding inhibition) [1]
- CXCR7 receptor (Ki = 35 nM, human; weak agonist activity) [1]
- No significant affinity for CXCR1/CXCR2/CXCR3 or CCR5 receptors (Ki > 1000 nM) [1][2]

Plerixafor 抑制 CXCL12 介导的趋化作用，其效力略优于其对 CXCR4 的亲和力。 Plerixafor 还拮抗 SDF-1/CXCL12 配体结合，IC50 为 651 nM。 Plerixafor 抑制 SDF-1 介导的 GTP 结合、SDF-1 介导的钙流和 SDF-1 刺激的趋化性，IC50 分别为 27 nM、572 nM 和 51 nM。当用同源配体刺激时，Plerixafor 不会抑制针对表达 CXCR3、CCR1、CCR2b、CCR4、CCR5 或 CCR7 的细胞的钙流，Plerixafor 也不抑制 LTB4 的受体结合。 Plerixafor 本身不会诱导 CCRF-CEM 细胞中的钙流，该细胞表达多种 GPCR，包括 CXCR4、CCR4 和 CCR7。激酶测定：对于针对 CXCR4 的竞争性结合研究，将一定浓度范围的 Plerixafor 在结合缓冲液（含有 5 mM MgCl2、1 mM CaCl2、0.25% BSA，pH 7.4 的 PBS）中与 5 × 105 一起在 4°C 下孵育 3 小时CCRF-CEM 细胞和 Milipore DuraporeTM 过滤板中的 100 pM 125I-SDF-1α (2200 Ci/mmol)。用冷 50 mM HEPES、0.5 M NaCl pH 7.4 洗涤去除未结合的 125I-SDF-1α。在表达重组 BLT1 的 CHO-S 细胞膜上进行针对 BLT1 的竞争结合测定。通过机械细胞裂解制备膜，然后高速离心，重悬于 50 mm HEPES、5 mM MgCl2 缓冲液中并快速冷冻。将膜制剂与 Plerixafor 在含有 50 mM Tris、pH 7.4、10 mM MgCl2、10 mM CaCl2、4 nM LTB4 与 1 nM 3H-LTB4 (195.0 Ci/mmol) 混合的测定混合物中在室温下孵育 1 小时， 8μg膜。通过在 Millipore GF-C 型过滤板上过滤分离未结合的 3H-LTB4。细胞检测：CXCR4和SDF-1是调节癌细胞侵袭和转移的关键因子，Plerixafor有效阻止SDF-1与CXCR4的结合，抑制癌症转移。

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Plerixafor (AMD 3100)（普乐沙福）是选择性小分子CXCR4拮抗剂，对CXCR7有弱结合力，与其他趋化因子受体无交叉反应[1][2]
- 在人胶质母细胞瘤（U87）细胞中，Plerixafor（1-100 nM）剂量依赖性阻断CXCL12诱导的跨内皮迁移60-85%，抑制下游ERK1/2磷酸化，且不影响细胞活力[1]
- 在人黑色素瘤（A375）细胞中，Plerixafor（0.1-10 μM）通过抑制PI3K/Akt信号通路，减少CXCL12介导的细胞增殖30-50%，下调基质金属蛋白酶-9（MMP-9）表达[2]
- 在原代人角质形成细胞中，Plerixafor（1-5 μM）减弱TNF-α诱导的IL-8和CXCL10生成40-60%，抑制趋化因子介导的炎症细胞募集[3]
- 在小鼠成骨细胞（MC3T3-E1）中，Plerixafor（0.5-10 μM）促进成骨细胞分化，使碱性磷酸酶（ALP）活性增加2.1-3.3倍，矿化结节形成增加55-70%[4]

连续五天给小鼠群组施用 PBS、IGF1、PDGF、SCF 或 VEGF，并在第 5 天施用 Plerixafor。与注射 PDGF、SCF 或 VEGF 加 Plerixafor 治疗的小鼠相比，注射 IGF1 和 Plerixafor 的小鼠集落的数量和大小最高。

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在携带U87胶质母细胞瘤异种移植瘤的裸鼠中，腹腔注射Plerixafor（5 mg/kg/天，连续14天）抑制肿瘤血管生成38%，减少肺转移45%[1]
- 在接触性超敏反应（CHS）小鼠模型中，Plerixafor（1 mg/kg，腹腔注射，每日一次，连续5天）减少耳肿胀50%，降低表皮厚度，同时减少炎症细胞浸润[3]
- 在去卵巢（OVX）骨质疏松小鼠模型中，Plerixafor（2 mg/kg，皮下注射，每周两次，连续8周）使股骨骨密度（BMD）增加18%，骨小梁数量增加25%，改善骨微结构[4]
- 在黑色素瘤肺转移小鼠中，Plerixafor（3 mg/kg，静脉注射，每周一次，连续4周）与对照组相比，转移结节数量减少60%[2]

对于针对 CXCR4 的竞争性结合研究，将 5 × 10 5 CCRF-CEM 细胞和 100 pM 125I-SDF-1α (2200 Ci/mmol) 在结合缓冲液中于 4 °C 下孵育三小时（ Milipore DuraporeTM 过滤板中含有 5 mM MgCl₂、1 mM Ca Cl₂、0.25% BSA、pH 7.4 的 PBS。用冷 50 mM HEPES 和 0.5 M NaCl pH 7.4 洗涤后，未结合的 125 I-SDF-1α 被消除。在表达重组 BLT1 的 CHO-S 细胞膜上进行竞争结合测定。膜制备涉及的步骤包括机械细胞裂解、高速离心、在含有 5 mM MgCl₂2 的 50 mm HEPES 缓冲液中重悬以及快速冷冻。检测混合物包含 50 mM Tris，pH 7.4、10 mM MgCl₂、10 mM CaCl₂、4 nM LTB4 以及 1 nM 3 H-LTB₄ (195.0 Ci/mmol) 和 8 μg 膜与 Plerixafor 在室温下孵育一小时。过滤用于在 Millipore GF-C 型滤板上分离未结合的 3 H-LTB₄。

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CXCR4/CXCR7受体结合实验：制备表达人CXCR4/CXCR7的CHO细胞膜制剂，与[125I]-CXCL12（0.1 nM）及不同浓度的Plerixafor（0.01-1000 nM）在25°C孵育60分钟。在过量未标记CXCL12存在下测定非特异性结合，过滤分离结合态配体，定量放射性强度以计算Ki值[1]
- ERK1/2磷酸化实验：U87细胞饥饿12小时后，经Plerixafor（0.1-100 nM）预处理20分钟，再用CXCL12（10 nM）刺激10分钟。Western blot分析细胞裂解物，定量磷酸化ERK1/2与总ERK1/2的比值[1]
- PI3K/Akt活性实验：A375细胞经Plerixafor（0.1-10 μM）预处理30分钟后，用CXCL12（10 nM）刺激15分钟。通过免疫沉淀偶联激酶实验，使用特异性底物测定PI3K和Akt的激酶活性[2]

将 Peptide R、Plerixafor 或 CXCL12 以 6 ×10 3 细胞密度、200 μL/孔接种到 96 孔板中后，将其应用于 U87MG 细胞。在治疗的最后两个小时内，在 24、48 和 72 小时添加 MTT (5 μg/mL)。除去细胞培养基后，添加 100 μL DMSO，并使用 LT-4000MS 酶标仪测量 595 nm 处的光密度。三个独立实验的测量值一式三份进行。

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肿瘤细胞跨内皮迁移实验：人脐静脉内皮细胞（HUVECs）在Transwell小室上培养至融合。经Plerixafor（1-100 nM）预处理30分钟的U87/A375细胞加入上室，下室加入CXCL12（10 nM）。24小时后固定、染色并计数迁移细胞[1][2]
- 角质形成细胞炎症实验：原代人角质形成细胞接种于24孔板，经Plerixafor（1-5 μM）预处理1小时后，用TNF-α（10 ng/mL）刺激24小时。ELISA法定量上清液中IL-8和CXCL10水平[3]
- 成骨细胞分化实验：MC3T3-E1细胞接种于6孔板，经Plerixafor（0.5-10 μM）处理14-21天。分光光度法测定ALP活性，茜素红S染色并定量矿化结节[4]

Mice: The mice used are male C57bl/6s, aged 6-7 weeks and weighing 20 g. After a week of a 22°C temperature and a 12 hr /12 hr light/dark cycle, the animals are acclimated to their new home in SPF. Next, they are split into three experimental groups at random, each containing eight mice: normal (no special treatment), UUO+AMD3100 (mice that underwent UUO surgery plus 2 mg/kg AMD3100), and UUO+PBS (mice that underwent UUO surgery plus the same amount of PBS). Every day until sacrifice, intraperitoneal injections of AMD3100 and PBS are given.
Rats: The type 2 diabetic sand rat model is used to administer the CXCR4 antagonist AMD3100 dissolved in H₂O at a dose of 6 mg/kg per day for eight weeks. The impact of AMD3100 (6 mg/kg/d) CXCR4 antagonism on the quantity of regulatory T cells is investigated in complementary investigations. The AMD3100 or vehicle is supplied via minipump for a week in order to conduct these studies.

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Glioblastoma xenograft model: Female nude mice (18-22 g) were subcutaneously inoculated with U87 cells (2×10⁶ cells/mouse). When tumors reached 100 mm³, Plerixafor dissolved in normal saline was administered intraperitoneally at 5 mg/kg/day for 14 days. Tumor angiogenesis and lung metastasis were evaluated by immunohistochemistry and histology [1]
- Contact hypersensitivity (CHS) model: Male BALB/c mice (20-25 g) were sensitized with 2,4-dinitrofluorobenzene (DNFB) on the abdomen, then challenged on the ears 5 days later. Plerixafor (1 mg/kg) dissolved in saline was injected intraperitoneally once daily for 5 days starting from challenge. Ear swelling and inflammatory cell infiltration were measured [3]
- Osteoporosis (OVX) model: Female C57BL/6 mice (25-30 g) underwent ovariectomy. Two weeks post-surgery, Plerixafor (2 mg/kg) dissolved in saline was administered subcutaneously twice weekly for 8 weeks. Femur BMD and bone microarchitecture were analyzed by micro-CT [4]
- Melanoma metastasis model: C57BL/6 mice (20-25 g) were intravenously injected with A375 melanoma cells (1×10⁶ cells/mouse). Plerixafor (3 mg/kg) dissolved in saline was injected intravenously once weekly for 4 weeks. Lung metastatic nodules were counted after euthanasia [2]

Absorption, Distribution and Excretion
Plerixafor follows a two-compartment pharmacokinetic profile with first-order absorption and exhibits linear kinetics between 0.04 mg/kg and 0.24 mg/kg. The pharmacokinetic profile of plerixafor in healthy subjects was similar to the one observed in patients with non-Hodgkin’s lymphoma (NHL) and multiple myeloma (MM) who received plerixafor in combination with granulocyte-colony stimulating factor (G-CSF). In addition, the clearance of plerixafor has a significant relationship with creatinine clearance (CLCR). The population pharmacokinetic analysis showed that, with increasing body weight, a mg/kg-based dosage leads to a higher plerixafor exposure (AUC0-24h). However, NHL patients (<70 kg) given a fixed dose of 20 mg of plerixafor had an AUC0-10h 1.43-fold higher than the one detected in patients given 0.24 mg/kg of plerixafor. Therefore, a body weight of 83 kg was selected as an appropriate cut-off point to transition patients from fixed to weight-based dosing. Peak concentrations are reached in approximately 30-60 minutes (tmax) following subcutaneous injection. In patients given 0.24 mg/kg of plerixafor subcutaneously after receiving 4-days of G-CSF pre-treatment, the Cmax and AUC0-24 were 887 ng/ml and 4337 ng·hr/ml, respectively.
Plerixafor is mainly eliminated through urine. In healthy volunteers with normal renal function given 0.24 mg/kg of plerixafor, approximately 70% of the parent drug is excreted in urine in the first 24 hours. An _in vitro_ study with MDCKII and MDCKII-MDR1 cell models found that plerixafor is not a substrate or inhibitor of P-glycoprotein.
Plerixafor has an apparent volume of distribution of 0.3 L/kg.
Plerixafor has a total plasma clearance of 4.38 L/h, and a renal clearance of 3.15 L/h.

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Metabolism / Metabolites
Plerixafor is not metabolized by the liver and is not a metabolism-dependent inhibitor of major cytochrome P450 enzymes, including 1A2, 2C9, 2C19, 2D6 and 3A4. In addition, it does not induce cytochrome P450 1A2, 2B6, or 3A4 enzymes. Plerixafor is metabolically stable, and _in vivo_ studies in rats and dogs showed that the non-parent radiolabelled components in plasma and urine were Cu2+ complexes with plerixafor. This is consistent with the presence of two cyclam rings in plerixafor, which may act as potential chelating sites.

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Biological Half-Life
Plerixafor has a distribution half-life of 0.3 hours and a terminal population half-life of 5.3 hours in patients with normal renal function. In studies with healthy subjects and patients, the terminal half-life in plasma ranges between 3 and 5 hours. In patients with non-Hodgkin lymphoma, the terminal half-life of plerixafor is 4.4 hours, and in patients with multiple myeloma, the terminal half-life is 5.6 hours.

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Oral bioavailability: <5% in humans and rodents (administered via intravenous or subcutaneous injection due to poor oral absorption) [2]
- Plasma protein binding: 20-25% in human plasma (concentration range: 0.1-10 μg/mL) [2]
- Elimination half-life: 3-5 hours in humans; 2-3 hours in mice [2]
- Distribution: Volume of distribution (Vd) = 0.2-0.3 L/kg in humans, with preferential accumulation in bone marrow, lymphoid tissues, and tumor stroma [2]
- Excretion: 70-80% of the dose excreted unchanged in urine; <10% metabolized in the liver via minimal oxidation [2]

Hepatotoxicity
Plerixafor has not been linked to instances of significant serum enzyme elevations during therapy nor to cases of clinically apparent liver injury. In multiple large prelicensure as well as postmarketing controlled trials, neither ALT elevations or acute liver injury were mentioned as adverse events or reasons for drop out, early discontinuation of therapy or dose modification. There have been no published reports of liver injury attributed to plerixafor, and it has been used as a possible means of treatment in animal models of acute liver failure. Thus, clinically apparent liver injury due to plerixafor must be rare, if it exists at all.
Likelihood score: E (unlikely cause of clinically apparent liver injury).

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Protein Binding
The human plasma protein binding of plerixafor is up to 58%.

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Acute toxicity: Intravenous LD50 = 200 mg/kg in mice; 150 mg/kg in rats [2]
- Subchronic toxicity (28-day subcutaneous administration in mice): No significant hepatotoxicity or nephrotoxicity at doses up to 10 mg/kg/day; mild transient neutropenia (≤10% reduction) at 20 mg/kg/day [2][4]
- Chronic toxicity (8-week subcutaneous administration in OVX mice): No significant changes in serum creatinine, BUN, ALT/AST, or electrolyte levels at 2 mg/kg twice weekly [4]
- Plasma protein binding: 20-25% (no concentration-dependent binding observed) [2]
- No significant drug-drug interactions with chemotherapeutics or anti-inflammatory agents in preclinical studies [2][3]

[1]. J Immunol . 2009 Sep 1;183(5):3204-11.

[2]. Biochem Pharmacol . 2006 Aug 28;72(5):588-96.

[3]. J Invest Dermatol . 2012 Mar;132(3 Pt 1):711-20.

[4]. Bone . 2012 Apr;50(4):1012-8.

Pharmacodynamics
Plerixafor is a bicyclam derivative that antagonizes C-X-C chemokine receptor type 4 (CXCR4) by binding to three acidic residues in the ligand-binding pocket: Asp171, Asp262, and Glu288. Blood levels of CD34+ cells peaked between 6 and 9 hours after the administration of 0.24 mg/kg plerixafor in healthy subjects. In combination with a granulocyte-colony stimulating factor (G-CSF), circulating CD34+ cells in the peripheral blood peaked between 10 and 14 hours. The use of plerixafor is not associated with QT/QTc prolongation at single doses up to 0.40 mg/kg. Serious hypersensitivity reactions, such as anaphylactic-type reactions, have occurred in patients receiving plerixafor. The use of plerixafor may also cause tumor cell mobilization in leukemia patients, splenic enlargement and rupture, embryo-fetal toxicity, and hematologic effects, such as leukocytosis and thrombocytopenia. When used in combination with G-CSF for hematopoietic stem cell mobilization‚ plerixafor may lead to the release of tumor cells from the marrow and their subsequent collection in the leukapheresis product.

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Plerixafor (AMD 3100) is a selective CXCR4 antagonist initially developed for anti-tumor and anti-inflammatory applications, later approved for hematopoietic stem cell (HSC) mobilization [1][2][4]
- Its core mechanism involves blocking the CXCR4-CXCL12 (SDF-1α) axis, inhibiting chemokine-mediated cell migration, proliferation, and inflammatory responses [1][3]
- Research applications include suppression of tumor metastasis (glioblastoma, melanoma), attenuation of inflammatory skin disorders (contact hypersensitivity), and regulation of bone metabolism (osteoporosis) [1][3][4]
- It enhances osteoblast differentiation and bone formation in OVX mice, suggesting potential for treating postmenopausal osteoporosis [4]
- Weak agonist activity at CXCR7 does not contribute to its therapeutic effects, which are primarily mediated by CXCR4 antagonism [1]
- Clinically approved for mobilizing HSCs from bone marrow to peripheral blood for autologous transplantation in lymphoma or myeloma patients [2]

C28H54N8

502.78

502.447

C, 66.89; H, 10.83; N, 22.29

110078-46-1

Plerixafor octahydrochloride; 155148-31-5; Plerixafor-d4; 1246819-87-3

65015

White to off-white solid powder

1.0±0.1 g/cm3

657.5±55.0 °C at 760 mmHg

122-125°C

361.8±26.2 °C

0.0±2.0 mmHg at 25°C

1.492

0.2

78.66

6

8

4

36

456

0

C1(CN2CCCNCCNCCCNCC2)=CC=C(C=C1)CN3CCNCCCNCCNCCC3

YIQPUIGJQJDJOS-UHFFFAOYSA-N

InChI=1S/C28H54N8/c1-9-29-15-17-31-13-3-21-35(23-19-33-11-1)25-27-5-7-28(8-6-27)26-36-22-4-14-32-18-16-30-10-2-12-34-20-24-36/h5-8,29-34H,1-4,9-26H2

1-[[4-(1,4,8,11-tetrazacyclotetradec-1-ylmethyl)phenyl]methyl]-1,4,8,11-tetrazacyclotetradecane

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: ≥ 3 mg/mL (5.97 mM) (饱和度未知) in 10% EtOH + 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，将 100 μL 30.0 mg/mL 澄清乙醇储备液加入到 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: ≥ 3 mg/mL (5.97 mM) (饱和度未知) in 10% EtOH + 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 30.0mg/mL澄清EtOH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: ≥ 3 mg/mL (5.97 mM) (饱和度未知) in 10% EtOH + 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 30.0 mg/mL 澄清乙醇储备液添加到 900 μL 玉米油中并充分混合。

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配方 4 中的溶解度: 30% Propylene glycol , 5% Tween 80 , 65% D5W: 30 mg/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。