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]

In vitro activity: Mavorixafor (AMD-070) is an effective and readily available oral CXCR4 antagonist. It inhibits the replication of T-tropic HIV-1 (NL4.3 strain) in MT-4 cells and PBMCs with an IC50 of 1 and 9 nM, respectively, and has an IC50 value of 13 nM against CXCR4 ¹²⁵I-SDF binding. Regarding other chemokine receptors (CCR1, CCR2b, CCR4, CCR5, CXCR1, and CXCR2), mavorixafor (AMD-070) exhibits no effect[1]. The growth, migration, and matrigel invasion of B88-SDF-1 cells are significantly suppressed by mavorixafor (AMD-070) at 6.6 µM[2].

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Mavorixafor (AMD070) inhibited the replication of T-tropic HIV-1 (NL4.3 strain) in MT-4 cells with an IC₅₀ of 2 nM and in PBMCs with an IC₅₀ of 26 nM; the compound was noncytotoxic to cells at concentrations exceeding 23 μM [1]
- Mavorixafor (AMD070) had no effect on the anchorage-dependent growth of B88-SDF-1 oral cancer cells, but significantly suppressed their anchorage-independent growth [3]
- Mavorixafor (AMD070) significantly inhibited the SDF-1/CXCR4-dependent migration and invasion of B88-SDF-1 oral cancer cells [3]

Mavorixafor (AMD-070) (2 mg/kg, p.o.) decreases the expression of human Alu DNA in mice and dramatically reduces the number of metastatic lung nodules in mice without causing a decrease in body weight[2].

The compounds are first preincubated in SUP-T1 T cells for 30 minutes on ice, with 1 serving as a control. Following this, the cells are washed with PBS containing 2% FCS and then incubated with PE-conjugated anti-CXCR4 mAb for an additional 30 minutes on ice. The cell samples are first fixed with 1% paraformaldehyde in PBS and then examined using an FACS Calibur flow cytometer after being cleaned in PBS. The average fluorescence intensity values are used to calculate the compounds' dose-dependent inhibitory effects on mAb binding.

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CXCR4 receptor binding assay [1]
: A radioligand binding assay was conducted using ¹²⁵I-SDF as the labeled ligand to evaluate the binding affinity of Mavorixafor (AMD070) to CXCR4. Cells expressing CXCR4 were incubated with varying concentrations of the compound along with a fixed amount of ¹²⁵I-SDF, and the amount of bound radioligand was measured after separating free and bound ligand. The IC₅₀ value for the inhibition of ¹²⁵I-SDF binding to CXCR4 by Mavorixafor (AMD070) was calculated from the dose-response curve.

On a 96-well plate, 5 × 10³ cells/well are seeded with DMEM containing 10% FCS. The cells are treated with or without 2 µM AMD3100 or 6.6 µM AMD-070 after a 24-hour period. An assay employing MTT is used to quantify the number of cells after 24 or 48 hours[2].

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HIV-1 replication inhibition assay in MT-4 cells and PBMCs [1]
: MT-4 cells and PBMCs were infected with T-tropic HIV-1 (NL4.3 strain) and then treated with different concentrations of Mavorixafor (AMD070). After a specific incubation period, the viral replication level was determined by measuring viral p24 antigen production or viral RNA levels. The IC₅₀ values for inhibiting HIV-1 replication in MT-4 cells and PBMCs were calculated based on the dose-response relationship. Additionally, cell viability was assessed using a cytotoxicity assay to determine the non-toxic concentration range of the compound.
- Anchorage-dependent and anchorage-independent growth assay of oral cancer cells [3]
: For the anchorage-dependent growth assay, B88-SDF-1 oral cancer cells were seeded in culture plates and treated with Mavorixafor (AMD070) at different concentrations; cell proliferation was measured by counting cells or using a colorimetric assay after a set incubation time. For the anchorage-independent growth assay, the cells were suspended in soft agar containing the drug and plated in culture dishes; the number of colonies formed was counted after several weeks of incubation to evaluate the inhibitory effect of the compound on colony formation.
- Cell migration and invasion assay [3]
: For the migration assay, B88-SDF-1 cells were seeded in the upper chamber of transwell inserts, and the lower chamber contained SDF-1 with or without Mavorixafor (AMD070); the number of cells that migrated through the membrane was counted after incubation. For the invasion assay, the transwell inserts were pre-coated with extracellular matrix, and the same procedure as the migration assay was followed to count the number of invasive cells, assessing the inhibitory effect of the drug on SDF-1/CXCR4-dependent invasion.

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, after a once-daily dose of 400 mg, the steady-state mean (CV%) Cmax was 3304 (58.6%) ng/mL, and the 0-24 hour AUC (AUC0-24h) was 13970 (58.4%) ng·h/mL. The pharmacokinetics of Mavorixafor are non-linear; the increases in Cmax and AUC0-24h are greater than dose-proportional across the dose range of 50 mg (0.125 times the recommended dose) to 400 mg. In healthy subjects, steady-state plasma concentrations of Mavorixafor are reached approximately after 9 to 12 days following administration of the highest approved recommended dose. At the highest approved recommended dose, the median (range) Tmax of Mavorixafor is 2.8 hours (1.9 to 4 hours). Food decreases Cmax and AUC.
Excretion Route
In healthy subjects, following a single oral dose of radiolabeled mavolixafor, 74.2% of the administered dose was recovered within a 240-hour collection period, with 61.0% of the radioactive material recovered in feces and 13.2% (3% unchanged) in urine.
Volume of Distribution
In adults with WHIM syndrome, the volume of distribution of mavolixafor is 768 L.
Clearance
In healthy subjects, following a single dose of 400 mg mavolixafor, the mean (coefficient of variation) apparent clearance was 62 L/h (40%). Mavolixafor exhibits at least partially nonlinear apparent clearance; however, this nonlinear clearance is not clinically significant at the approved recommended dose.
Protein Binding
In vitro studies have shown that mavolixafor binds to human plasma proteins >93%.
Metabolism/Metabolites
Mavolixafor is primarily metabolized via CYP3A4, and secondarily via CYP2D6.
Biological Half-Life
In healthy subjects, the mean (CV%) terminal half-life after a single dose of 400 mg mavolixafor is 82 hours (34%).
Mavorixafor (AMD070) is a small molecule drug with high oral bioavailability; good oral bioavailability was observed in both rats and dogs [1]
- In CD-1 mice, after oral administration of 400 μg/mouse of Mavorixafor (AMD070), the drug was detected in lung tissue, and its concentration varied over time. EC₉₀ (44 ng/mL) was used as the reference threshold for pharmacodynamic activity [2]
- In C57BL/6 mice, after intraperitoneal injection of Mavorixafor (AMD070), the drug was distributed in plasma, liver, and lung tissue, and its concentration was detected at different time points [2]

Mavorixafor (AMD070) showed no cytotoxicity to MT-4 cells and PBMCs at in vitro concentrations exceeding 23 μM[1]
Effects during pregnancy and lactation
◉ Overview of use during lactation
There is currently no information regarding the use of mavorixafor during lactation. The manufacturer recommends against breastfeeding during treatment and for three weeks after the last dose.
◉ Effects on breastfed infants
No published information was found as of the revision date.
◉ Effects on lactation and breast milk
No published information was 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 drug. Mavorixafor is a small molecule candidate drug belonging to a new class of anti-HIV drugs called viral entry (fusion) inhibitors. Currently, only one FDA-approved viral entry inhibitor—enfuvirdi (fuzeon)—is used to treat HIV infection. Several experimental viral entry inhibitors, including mavorixafor, are in early clinical trials. Mavorixafor is a selective CXCR4 receptor allosteric antagonist that inhibits HIV entry into and infection of healthy cells. Mavorixafor is an orally bioavailable CXC chemokine receptor 4 (CXCR4) inhibitor with potential anti-tumor and immune checkpoint inhibitory activity. After administration, mavorixafor selectively binds to CXCR4, preventing CXCR4 from binding to its ligand, stromal cell-derived factor 1 (SDF-1 or CXCL12). This inhibits receptor activation, leading to reduced proliferation and migration of CXCR4-overexpressing tumor cells. Furthermore, inhibiting CXCR4 prevents the recruitment of regulatory T cells and myeloid-derived suppressor cells (MDSCs) to the tumor microenvironment, thereby eliminating CXCR4-mediated immunosuppression and activating cytotoxic T lymphocyte-mediated immune responses against cancer cells. The G protein-coupled receptor CXCR4 is highly expressed in various tumor cell types, inducing the recruitment of immunosuppressive cells to the tumor microenvironment, inhibiting immune surveillance, and promoting tumor angiogenesis and tumor cell proliferation. It is also a co-receptor for HIV entry into T cells.

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Drug Indications
It has been investigated for the treatment of HIV infection.

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Mechanism of Action
Chemokine receptors expressed on the surface of immune cells are known to play a crucial role in viral infection and transmission. CXCR4 and another chemokine receptor, CCR5, are involved in HIV infection. The process of HIV entry into cells begins with the binding of viral envelope glycoproteins to the CD4 receptor and one of only two chemokine receptors, ultimately leading to viral membrane fusion with the cell membrane. Viral entry provides a novel target for anti-HIV therapy. To date, at least three subclasses of HIV entry/fusion inhibitors have emerged: 1. CD4 binding or attachment – targeting the initial recognition and binding of viral glycoprotein gp120 to cell surface CD4 antigens. 2. Chemokine co-receptor binding – targeting viral binding to CCR5 or CXCR4 co-receptors. 3. Fusion inhibition – targeting viral glycoprotein gp41 to inhibit viral fusion with cells. Different HIV strains prefer different receptors or may use either receptor to infect cells. 35% of strains use both CXCR4 and CCR5. 5% of strains use only CXCR4. 60% of strains use only CCR5. Different levels of CXCR4 and CCR5-binding virus may exist in infected individuals. Virus using CXCR4 can independently predict CD4 decline and HIV clinical progression, and is associated with early death. Pharmacodynamics AMD-070 is a small molecule drug candidate belonging to a novel class of investigational anti-HIV drugs called entry (fusion) inhibitors. Approximately 76% of HIV-infected individuals with measurable viral loads are infected with viral strains resistant to one or more antiretroviral drugs, limiting their treatment options. Unlike many existing HIV drugs that work after the virus infects healthy cells, AMD-070 blocks viral replication by blocking entry into healthy cells. AMD-070 targets the CXCR4 receptor on the surface of the HIV virus, preventing the virus from entering and infecting healthy cells. AMD-070 specifically targets the CXCR4 receptor, and in vitro studies have shown that it does not interact with other chemokine receptors. In vitro studies have shown that AMD-070 potently inhibits infection by all viruses that utilize the CXCR4 receptor (including viruses that utilize only the CXCR4 receptor and/or viruses that utilize both CXCR4 and CCR5 receptors). AMD-070 has oral bioavailability in animals. Its pharmacokinetic and toxicological characteristics are suitable for oral administration. In vitro studies have shown that AMD-070 has an additive or synergistic effect when used in combination with other known anti-HIV drugs. AMD-070 is active against HIV strains that are resistant to existing antiretroviral therapies and utilize the CXCR4 receptor in vitro. It has potent anti-HIV activity against both laboratory strains and clinical isolates that utilize the CXCR4 receptor.

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Mavorixafor (AMD070) is the first small molecule oral bioavailable CXCR4 antagonist 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 pathway but does not effectively reduce extracellular matrix deposition in pulmonary or hepatic fibrosis; however, it improves mortality from bleomycin (BLM)-induced lung injury, possibly by inhibiting early inflammatory responses and/or vascular leakage [2]. Mavorixafor (AMD070) is a minimally invasive alternative to AMD3100 (a subcutaneously injected CXCR4 antagonist) for inhibiting CXCR4-related metastases in 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

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

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

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

5%DMSO + 40%PEG300 + 5%Tween 80 + 50%ddH2O: 3.5mg/ml (10.02mM) (Please use freshly prepared in vivo formulations for optimal results.)