¹²⁵I-CXCL12-CXCR4 ( IC50 = 44 nM ); ¹²⁵I-CXCL12-CXCR7; HIV-1 ( EC50 = 1-10 nM ); HIV-2 ( IC50 = 1-10 nM )

In diabetic mice, a single topical application of Plerixafor increases the production of cytokines, mobilizes bone marrow EPCs, and activates fibroblasts, monocytes/macrophages, and fibroblasts, thereby increasing angiogenesis and vasculogenesis.[3] Mice are given PBS, IGF1, PDGF, SCF, or VEGF for five days in a row, and Plerixafor on the fifth day. Compared to groups treated with PDGF, SCF, and VEGF in addition to Plerixafor, mice injected with IGF1 plus Plerixafor exhibited the largest colonies in terms of both number and size.[4]

For the competition binding studies against CXCR4, 5 × 10⁵ CCRF-CEM cells and 100 pM 125I-SDF-1α (2200 Ci/mmol) are incubated for three hours at 4 °C in binding buffer (PBS containing 5 mM MgCl₂, 1 mM Ca Cl₂, 0.25% BSA, pH 7.4) in Milipore DuraporeTM filter plates. After washing with cold 50 mM HEPES and 0.5 M NaCl pH 7.4, unbound ¹²⁵I-SDF-1α is eliminated. On membranes from CHO-S cells expressing recombinant BLT1, the competition binding assay is carried out. Mechanical cell lysis, high-speed centrifugation, resuspension in 50 mm HEPES buffer containing 5 mM MgCl₂2, and flash freezing are the steps involved in the preparation of the membranes. The assay mixture comprising 50 mM Tris, pH 7.4, 10 mM MgCl₂, 10 mM CaCl₂, 4 nM LTB4 combined with 1 nM ³H-LTB₄ (195.0 Ci/mmol) and 8 μg membrane is incubated with Plerixafor for one hour at room temperature. Filtration is used to separate the unbound ³H-LTB₄ on Millipore Type GF-C filter plates.

Peptide R, Plerixafor, or CXCL12 are applied to U87MG cells after they are seeded at a density of 6 ×10³ cells in 200 μL/well in 96-well plates. During the last two hours of treatment, MTT (5 μg/mL) is added at 24, 48, and 72 hours. Following the removal of the cell medium, 100 μL of DMSO is added, and an LT-4000MS Microplate Reader is used to measure the optical densities at 595 nm. Three separate experiments' worth of measurements are taken in triplicate.

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.

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%.

[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.

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

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)

Solubility in Formulation 1: ≥ 3 mg/mL (5.97 mM) (saturation unknown) in 10% EtOH + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear EtOH stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 3 mg/mL (5.97 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 3 mg/mL (5.97 mM) (saturation unknown) in 10% EtOH + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix well.

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Solubility in Formulation 4: 30% Propylene glycol , 5% Tween 80 , 65% D5W: 30 mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)