¹²⁵I-SDF-CXCR4 ( IC50 = 13 nM ); HIV-1 (NL4.3 strain) ( IC50 = 1 nM ); HIV-1 (NL4.3 strain) ( IC50 = 9 nM ); HIV-1 (NL4.3 strain) ( IC50 = 3 nM ); HIV-1 (NL4.3 strain) ( IC50 = 26 nM )
Mavorixafor (AMD070) targets CXCR4 (IC₅₀ = 13 nM in CXCR4 ¹²⁵I-SDF inhibition binding assay) [1]
Mavorixafor (AMD070) acts as an antagonist of the CXCL12/CXCR4 signaling axis [2]

体外活性：AMD-070 对 MT-4 细胞中的 HIV-1、HIV-1 IIIb X4 株有活性，AMD-070 的 IC50 值高出 9 倍（0.009 μM vs 0.001 μM）和 8.7-与 MT-4 细胞相比，PBMC 中的浓度高出一倍（0.003 μM vs 0.026 μM）。 AMD-070具有抗病毒能力，IC50值为15.5 nM。激酶测定：首先将 SUP-T1 T 细胞与化合物（以 1 作为对照）在冰上预孵育 30 分钟，用含 2% FCS 的 PBS 洗涤，并与 PE 缀合的抗 CXCR4 mAb 在冰上孵育 30 分钟。用 PBS 洗涤后，用 1% 多聚甲醛的 PBS 溶液固定细胞样品，并在 FACS Calibur 流式细胞仪上进行分析。使用平均荧光强度值确定化合物对 mAb 结合的剂量依赖性抑制作用。细胞测定：激活的细胞（PHA刺激的母细胞）用PBS洗涤3次，完成病毒感染。 HIV 感染或模拟感染的 PHA 刺激的母细胞在 25 U/mL IL-2 和不同浓度的化合物存在下培养。第10天收集上清液，并通过p24病毒Ag ELISA试剂盒分析培养物上清液中的HIV-1核心抗原。如前所述进行MT-4细胞中HIV-1复制的抑制。抗 HIV-1 活性和细胞毒性测量同时进行。它们基于感染 HIV-1 的 MT-4 细胞在不同浓度的测试化合物存在下的活力。 ICsub>50定义为抑制50%的病毒感染细胞抵抗病毒细胞病变所需的浓度。

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Mavorixafor (AMD070)在MT-4细胞中抑制T嗜性HIV-1（NL4.3株）复制的IC₅₀为2 nM，在PBMCs中为26 nM；该化合物在浓度超过23 μM时，对细胞无细胞毒性[1]
- Mavorixafor (AMD070)对B88-SDF-1口腔癌细胞的贴壁依赖性生长无影响，但能显著抑制其非贴壁依赖性生长[3]
- Mavorixafor (AMD070)可显著抑制B88-SDF-1口腔癌细胞由SDF-1/CXCR4介导的迁移和侵袭能力[3]

AMD-070 在大鼠和狗中显示出良好的口服生物利用度。清除率取决于物种，与大鼠相比，AMD-070 在狗体内的清除率较低。

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在CD-1小鼠中，经口给予200 μg/只或400 μg/只剂量的Mavorixafor (AMD070)，可显著增加血液中白细胞的动员，且主要表现为淋巴细胞数量增多；药物同时升高白细胞计数，对红细胞和血小板无显著影响[2]
- 在博来霉素（BLM）诱导的小鼠肺纤维化模型中，Mavorixafor (AMD070)处理未能缓解肺组织的炎症（H&E染色）和细胞外基质沉积（Masson三色染色），但可显著降低BLM诱导的肺损伤相关死亡率[2]
- 在四氯化碳（CCl₄）诱导的C57BL/6小鼠肝纤维化模型中，Mavorixafor (AMD070)对肝纤维化（天狼星红染色）无改善作用，也未改变肝脏中Acta2和Col1a1的转录水平及血清AST水平[2]
- 裸鼠中每日经口给予Mavorixafor (AMD070)，可显著抑制B88-SDF-1口腔癌细胞的肺转移[3]

这些化合物首先在冰上的 SUP-T1 T 细胞中预孵育 30 分钟，其中 1 个作为对照。随后，用含有 2% FCS 的 PBS 洗涤细胞，然后与 PE 缀合的抗 CXCR4 mAb 在冰上再孵育 30 分钟。细胞样品首先用 PBS 中的 1% 多聚甲醛固定，然后在 PBS 中清洗后使用 FACS Calibur 流式细胞仪进行检查。平均荧光强度值用于计算化合物对 mAb 结合的剂量依赖性抑制作用。

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CXCR4受体结合实验 [1]
: 采用放射性配体结合实验，以¹²⁵I-SDF作为标记配体，评估Mavorixafor (AMD070)与CXCR4的结合亲和力。将表达CXCR4的细胞与不同浓度的该化合物及定量的¹²⁵I-SDF共同孵育，分离游离和结合的配体后检测结合的放射性配体量，根据剂量-反应曲线计算出Mavorixafor (AMD070)抑制¹²⁵I-SDF与CXCR4结合的IC₅₀值。

在 96 孔板上，以 5 × 10³ 细胞/孔接种含有 10% FCS 的 DMEM。 24 小时后，用或不用 2 µM AMD3100 或 6.6 µM AMD-070 处理细胞。采用 MTT 的测定用于量化 24 或 48 小时后的细胞数量[2]。

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MT-4细胞和PBMCs中HIV-1复制抑制实验 [1]
: 将MT-4细胞和PBMCs感染T嗜性HIV-1（NL4.3株）后，用不同浓度的Mavorixafor (AMD070)处理。孵育特定时间后，通过检测病毒p24抗原产生量或病毒RNA水平确定病毒复制水平，依据剂量-反应关系计算药物在MT-4细胞和PBMCs中抑制HIV-1复制的IC₅₀值。同时采用细胞毒性实验评估细胞活力，确定化合物的无毒浓度范围。
- 口腔癌细胞贴壁依赖性和非贴壁依赖性生长实验 [3]
: 贴壁依赖性生长实验中，将B88-SDF-1口腔癌细胞接种于培养板，加入不同浓度的Mavorixafor (AMD070)，孵育一定时间后通过细胞计数或比色法检测细胞增殖情况。非贴壁依赖性生长实验中，将细胞悬浮于含药软琼脂中并接种于培养皿，培养数周后计数形成的集落数，评估化合物对集落形成的抑制作用。
- 细胞迁移和侵袭实验 [3]
: 迁移实验中，将B88-SDF-1细胞接种于transwell小室上室，下室加入含或不含Mavorixafor (AMD070)的SDF-1，孵育后计数穿过膜的细胞数。侵袭实验中，先对transwell小室进行细胞外基质包被，再按迁移实验流程操作并计数侵袭细胞数，以此评估药物对SDF-1/CXCR4介导的侵袭的抑制作用。

Mice: In a pathogen-free environment, BALB/c nude mice are raised. As soon as the mice reach eight weeks of age, the experiments begin. In summary, 1×10⁶ nude mice have their blood vessels inoculated with the cells. Day 49 is the time of the mice's sacrifice. H&E staining is used to determine whether distant metastases are present or absent. The mice are given 0.2 mL of saline as a vehicle or the same volume of Mavorixafor (AMD-070) (2 mg/kg) orally every day as part of their experimental chemotherapy treatment[2].

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Pharmacokinetic study in CD-1 and C57BL/6 mice [2]
: CD-1 mice were given a single oral dose of Mavorixafor (AMD070) at 400 μg/animal, and lung tissue samples were collected at different time points to measure the drug concentration using a quantitative analytical method. C57BL/6 mice received an intraperitoneal (IP) injection of the drug, and plasma, liver, and lung samples were collected at various time points to determine the drug concentration in these tissues, with the mean concentration and standard error of the mean (SEM) calculated for each time point.
- Leukocyte mobilization study in mice [2]
: CD-1 mice were administered Mavorixafor (AMD070) orally at doses of 200 μg/mouse or 400 μg/mouse, and blood samples were collected at different time points to count WBCs, RBCs, platelets, and perform differential cell counting (lymphocytes, neutrophils, monocytes, eosinophils) using standard hematological methods. C57BL/6 mice were given an IP injection of 400 μg/mouse of the drug, and blood cell counts were conducted following the same protocol to assess leukocyte mobilization.
- Bleomycin-induced pulmonary fibrosis model in mice [2]
: CD-1 mice were administered bleomycin (BLM) to induce pulmonary fibrosis, and Mavorixafor (AMD070) was given orally at a specified dose starting from a certain time after BLM treatment. Control groups received PBS plus acetate buffer or BLM plus acetate buffer. At the end point, lung tissues were collected for H&E staining (to evaluate inflammation) and Masson’s Trichrome staining (to assess fibrosis), and mouse survival was monitored over time to analyze the effect of the drug on mortality.
- Carbon tetrachloride-induced hepatic fibrosis model in mice [2]
: C57BL/6 mice were treated with carbon tetrachloride (CCl₄) to induce hepatic fibrosis, and Mavorixafor (AMD070) was administered intraperitoneally at a set dose. Control groups received oil plus PBS or CCl₄ plus PBS. After the treatment period, liver tissues were collected for Picosirius red staining to quantify fibrosis, and the transcription levels of Acta2 and Col1a1 were measured by qPCR; serum AST levels were also determined to evaluate liver injury.
- Oral cancer metastasis model in nude mice [3]
: B88-SDF-1 oral cancer cells were injected into nude mice to establish a metastasis model. Mavorixafor (AMD070) was administered orally on a daily basis at a specific dose starting from the day of tumor cell injection or a predetermined time point. Control mice received a vehicle solution. After a certain period, the mice were euthanized, and lung tissues were examined to count the number of metastatic nodules, evaluating the inhibitory effect of the drug on lung metastasis.

Absorption
In adults with WHIM syndrome, the mean (CV%) Cmax at steady-state is 3304 (58.6%) ng/mL and the AUC from 0 to 24 hours (AUC0-24h) is 13970 (58.4%) ngxh/mL following 400 mg once daily. Mavorixafor demonstrates nonlinear pharmacokinetics with greater than dose-proportional increases in Cmax and AUC0-24h over a dose range of 50 mg (0.125 times the recommended dosage) to 400 mg. Mavorixafor steady-state is reached after approximately 9 to 12 days at the highest approved recommended dosage in healthy subjects. Mavorixafor median (range) Tmax is 2.8 hours (1.9 to 4 hours) at the highest approved recommended dosage. Food decreases Cmax and AUC.

Route of Elimination
After a single oral dose of radiolabeled mavorixafor, 74.2% of the administered dose was recovered out of which 61.0% of administered radioactivity was recovered in feces and 13.2% (3% unchanged) was recovered in the urine over the 240-hour collection period in healthy subjects.

Volume of Distribution
Mavorixafor volume of distribution was 768 L in adults with WHIM syndrome.

Clearance
The mean (CV%) apparent clearance was 62 L/h (40%) following a single-dose administration of mavorixafor 400 mg in healthy subjects. Mavorixafor exhibits at least partial nonlinear apparent clearance; however, this is not clinically significant at the approved recommended dosage.

Protein Binding
In vitro, mavorixafor is >93% bound to human plasma proteins.

Metabolism / Metabolites
Mavorixafor is metabolized by CYP3A4 and, to a lesser extent, CYP2D6.

Biological Half-Life
The mean (CV%) terminal half-life was 82 h (34%) following a single-dose administration of mavorixafor 400 mg in healthy subjects.

Mavorixafor (AMD070) is an orally bioavailable small molecule; good oral bioavailability was observed in both rats and dogs [1]
- In CD-1 mice, after oral administration of Mavorixafor (AMD070) at 400 μg/animal, the drug was detected in the lung tissue, and its concentration varied over time, with the EC₉₀ (44 ng/mL) used as a reference threshold for pharmacodynamic activity [2]
- In C57BL/6 mice, following intraperitoneal administration of Mavorixafor (AMD070), the drug was distributed in plasma, liver, and lung tissues, with measurable concentrations at different time points [2]

Mavorixafor (AMD070) was noncytotoxic to MT-4 cells and PBMCs at concentrations exceeding 23 μM in vitro [1]

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the use of mavorixafor during breastfeeding. The manufacturer recommends not breastfeeding treatment and for three weeks after the final dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

[1]. Discovery of novel small molecule orally bioavailable C-X-C chemokine receptor 4 antagonists that are potent inhibitors of T-tropic (X4) HIV-1 replication. J Med Chem. 2010 Apr 22;53(8):3376-88.

[2]. Impact of a CXCL12/CXCR4 Antagonist in Bleomycin (BLM) Induced Pulmonary Fibrosis and Carbon Tetrachloride (CCl4) Induced Hepatic Fibrosis in Mice. PLoS One. 2016 Mar 21;11(3):e0151765.

[3]. Effect of a novel orally bioavailable CXCR4 inhibitor, AMD070, on the metastasis of oral cancer cells. Oncol Rep. 2018 Jul;40(1):303-308.

AMD 070 is an aminoquinoline.
Mavorixafor is a small molecule drug candidate that belongs to a new investigational class of anti-HIV drugs known as entry (fusion) inhibitors. Currently there is only one FDA-approved entry inhibitor, enfuvirtide (Fuzeon), that is available for the treatment of HIV infection. Several experimental entry inhibitors are now in early stage testing, including mavorixafor. Mavorixafor is a selective allosteric antagonist of the CXCR4 receptor on HIV, preventing the virus from entering and infecting healthy cells.
Mavorixafor is an orally bioavailable inhibitor of C-X-C chemokine receptor type 4 (CXCR4), with potential antineoplastic and immune checkpoint inhibitory activities. Upon administration, mavorixafor selectively binds to CXCR4 and prevents the binding of CXCR4 to its ligand, stromal cell-derived factor 1 (SDF-1 or CXCL12). This inhibits receptor activation and results in decreased proliferation and migration of CXCR4-overexpressing tumor cells. In addition, inhibition of CXCR4 prevents the recruitment of regulatory T-cells and myeloid-derived suppressor cells (MDSCs) to the tumor microenvironment, thereby abrogating CXCR4-mediated immunosuppression and enabling the activation of a cytotoxic T-lymphocyte-mediated immune response against cancer cells. The G protein-coupled receptor CXCR4, which is upregulated in several tumor cell types, induces the recruitment of immunosuppressive cells in the tumor microenvironment, suppresses immune surveillance, and promotes tumor angiogenesis and tumor cell proliferation. It is also a co-receptor for HIV entry into T-cells.

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Drug Indication
Investigated for use/treatment in HIV infection.

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Mechanism of Action
Chemokine receptors expressed on the surface of immune cells are known to play a critical role in virus infection and transmission. CXCR4, and another chemokine receptor CCR5, are involved in HIV infection. The process of HIV entry begins with binding of the viral envelope glycoprotein to both the CD4 receptor and one of only two chemokine receptors, and ends with fusion of viral and cell membranes. Viral entry provides novel therapeutic targets against HIV. To date, at least 3 sub classes of HIV viral entry/fusion inhibitors have emerged: 1. CD4 binding or attachment - targets initial recognition and binding of the viral glycoprotein gp120 to the cell-surface CD4 antigen. 2. Chemokine co-receptor binding - targets binding of virus to the CCR5 or CXCR4 co-receptor. 3. Fusion Inhibition - targets the viral glycoprotein gp41 inhibiting the fusion of virus with the cell. Different strains of HIV prefer one receptor or the other, or may use either receptor to infect cells. * 35% of strains use both CXCR4 and CCR5 * 5% of strains are pure CXCR4 using * 60% of strains are pure CCR5 using * An infected individual may harbor different levels of both CXCR4 and CCR5 using virus * CXCR4 using virus independently predicts CD4 decline and HIV clinical progression and is associated with earlier mortality

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Pharmacodynamics
AMD-070 is a small molecule drug candidate that belongs to a new investigational class of anti-HIV drugs known as entry (fusion) inhibitors. Approximately 76% of HIV-patients with measurable viral load are infected with a strain of virus that is resistant to one or more classes of antiretroviral agents, thus reducing treatment options. Unlike many existing HIV drugs that target the virus after it has infected a healthy cell, AMD-070 blocks the virus from entering a healthy cell, thus preventing the replication process. AMD-070 targets the CXCR4 receptor on HIV and prevents the virus from entering and infecting healthy cells. * AMD-070 is specific for the CXCR4 receptor and does not interact with any other chemokine receptors in vitro * AMD-070 strongly inhibits viral infection by all CXCR4 using virus (including virus using CXCR4 alone and/or virus using CXCR4 and CCR5) in vitro * AMD-070 is orally bioavailable in animals * Suitable PK and toxicity profile for oral dosing * AMD-070 shows additive or synergistic effects in vitro in combination with other known anti-HIV agents * AMD-070 is active against CXCR4 using HIV strains that are resistant to existing antiretroviral therapies in vitro * Potent anti-HIV activity against CXCR4-using laboratory strains and clinical isolates

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Mavorixafor (AMD070) is the first small molecule orally bioavailable CXCR4 antagonist developed for the treatment of HIV-1 infection [1]
- Mavorixafor (AMD070) exerts its anti-HIV-1 effect by antagonizing the CXCR4 receptor, thereby inhibiting the entry of T-tropic (X4) HIV-1 into host cells [1]
- In fibrotic disease models, Mavorixafor (AMD070) modulates the CXCL12/CXCR4 signaling axis but does not effectively reduce extracellular matrix deposition in pulmonary or hepatic fibrosis; however, it improves mortality in BLM-induced pulmonary injury, likely by dampening the early inflammatory response and/or vascular leakage [2]
- Mavorixafor (AMD070) is a minimally invasive alternative to AMD3100 (a subcutaneous CXCR4 antagonist) for inhibiting CXCR4-related metastasis of head and neck cancer [3]

C₂₁H₂₇N₅

349.47

349.226

C, 72.17; H, 7.79; N, 20.04

558447-26-0

Mavorixafor trihydrochloride; 2309699-17-8; 880549-30-4

11256587

White to gray solid powder

1.2±0.1 g/cm3

597.0±50.0 °C at 760 mmHg

108-110ºC

314.9±30.1 °C

0.0±1.7 mmHg at 25°C

1.656

2.78

70.83

2

4

7

26

431

1

NCCCCN(CC1=NC2=C(N1)C=CC=C2)[C@@H]3C4=C(CCC3)C=CC=N4

WVLHHLRVNDMIAR-IBGZPJMESA-N

InChI=1S/C21H27N5/c22-12-3-4-14-26(15-20-24-17-9-1-2-10-18(17)25-20)19-11-5-7-16-8-6-13-23-21(16)19/h1-2,6,8-10,13,19H,3-5,7,11-12,14-15,22H2,(H,24,25)/t19-/m0/s1

N'-(1H-benzimidazol-2-ylmethyl)-N'-[(8S)-5,6,7,8-tetrahydroquinolin-8-yl]butane-1,4-diamine

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。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)