Autotaxin (IC50 = 2.8 nM)
PF-8380 is a selective inhibitor of autotaxin (ATX, also named ectonucleotide pyrophosphatase/phosphodiesterase 2, ENPP2); the IC50 for recombinant human ATX is 25 nM, and the Ki value for human ATX is 10 nM [1]
PF-8380 has no significant inhibitory activity (IC50 > 10 μM) against other phosphodiesterases (PDE1-PDE11) or lysophospholipases [1]

Additionally, PF-8380 inhibits rat autotaxin, a substrate for FS-3, with an IC50 of 1.16 nM. When fetal fibroblast-produced enzymes were combined with lysophosphatidylcholine (LPC) as a substrate, PF-8380's efficacy remained intact. When human whole blood was treated with PF-8380 for two hours at an IC50 of 101 nM, autocrine motility factors were suppressed [1]. Lysophospholipase D (lysoPLD) activity is exhibited by the enzyme autotaxin (ATX), which catalyzes the conversion of lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA). After applying 1 μM PF-8380 as a pretreatment to GL261 and U87-MG cells, they were exposed to 4 Gy of radiation, which led to a decrease in clone survival, reduced migration (33% in GL261; P=0.002 and 17.9% in U87-MG; P=0.012), decreased invasion (35.6% in GL261; P=0.0037; 31.8% in U87-MG; P=0.002), and attenuate radiation-induced Akt phosphorylation [2].

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1. ATX activity inhibition: PF-8380 (0.1–100 nM) concentration-dependently inhibited recombinant human and mouse ATX activity in vitro, with complete inhibition of human ATX observed at 100 nM; it also reduced lysophosphatidic acid (LPA) production from lysophosphatidylcholine (LPC) in human plasma (IC50 = 30 nM) [1]
2. Glioblastoma cell lines: PF-8380 (1–50 μM) dose-dependently inhibited the proliferation of human (U87, U251) and murine (GL261) glioblastoma cell lines with IC50 values of 12 μM (U87), 15 μM (U251), and 10 μM (GL261) after 72-hour treatment; it enhanced the radiosensitivity of these cells, with the radiation enhancement ratio (RER) of 1.8 for U87 cells at 5 μM [2]
3. Glioblastoma clonogenicity and apoptosis: PF-8380 (5 μM) reduced the clonogenic formation of U87 cells by 60% and increased radiation-induced apoptosis (Annexin V+/PI+ cells from 12% to 38% after 4 Gy irradiation); western blot showed upregulation of cleaved caspase-3 and downregulation of anti-apoptotic Bcl-2 [2]
4. Lung allograft fibrosis-related cells: PF-8380 (10 μM) inhibited LPA-induced β-catenin phosphorylation (Ser552) and nuclear translocation in human lung fibroblasts (HLFs), reduced the expression of fibrotic markers (α-SMA, collagen I) by 55% and 48% (qPCR), and suppressed HLF proliferation (IC50 = 8 μM) [3]
5. LPA signaling inhibition: PF-8380 (5–20 μM) blocked LPA-induced ERK1/2 and AKT phosphorylation in glioblastoma and lung fibroblast cells, indicating inhibition of the LPA-Gi/PI3K/ERK signaling axis [2][3]

PF-8380's pharmacokinetic properties were assessed over the course of 24 hours at intravenous dosages of 1 mg/kg and oral doses ranging from 1 to 100 mg/kg. PF-8380 has an effective t1/2 of 1.2 hours, a steady-state distribution volume of 3.2 L/kg, and an average clearance rate of 31 mL/min/kg. There is moderate oral bioavailability, with a range of 43% to 83%. As single oral dosages are increased, plasma concentrations rise as well; however, the rate of increase in Cmax is less than proportional to doses of 10 to 100 mg/kg but about proportionate to doses of 1 to 10 mg/kg. Up to 100 mg/kg, exposure to PF-8380 is roughly dose-proportional and linear, as indicated by the area under the curve. The amounts of plasma C16:0, C18:0, and C20:0 LPA were tested right away following collection. With the 3 mg/kg dose, the largest drop in LPA levels was seen at 0.5 hours, and within 24 hours, all LPAs were returned to or exceeded baseline [1]. Tumor-associated vascularity increased by a moderate 20% after treatment with 10 mg/kg PF-8380 (P=0.497). 45 minutes prior to 4 Gy irradiation, PF-8380 treatment decreased vascularity in mice treated relative to controls by approximately 48% (P=0.031) and by 65% (P=0.011) in mice treated with radiation alone[2].

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1. Rat inflammatory model: In carrageenan-induced rat paw edema model, oral administration of PF-8380 (30 mg/kg) reduced plasma LPA levels by 70% and paw LPA levels by 65% at 4 hours post-administration; paw swelling was decreased by 50% compared with vehicle-treated rats, and myeloperoxidase (MPO) activity (neutrophil infiltration) was reduced by 60% [1]
2. Glioblastoma xenograft model: In U87 glioblastoma xenografts (nude mice), intraperitoneal injection of PF-8380 (20 mg/kg, once daily) combined with fractionated radiation (2 Gy/day for 5 days) achieved 80% tumor growth inhibition (TGI), compared with 35% TGI for radiation alone and 25% TGI for PF-8380 monotherapy; the combination also extended median survival from 32 days (radiation alone) to 58 days [2]
3. Mouse lung allograft fibrosis model: In a murine orthotopic lung transplantation model, oral administration of PF-8380 (15 mg/kg, twice daily) for 28 days reduced lung allograft fibrosis score from 3.5 to 1.2 (scale 0–4), decreased collagen deposition by 62% (Masson’s trichrome staining), and inhibited β-catenin nuclear accumulation in lung tissues by 70% (immunofluorescence) [3]
4. LPA level modulation in vivo: PF-8380 (30 mg/kg, oral) reduced LPA levels in bronchoalveolar lavage fluid (BALF) of lung transplant mice by 55% and downregulated the expression of LPA receptors (LPA1, LPA2) in lung tissues [3]

ATX ELISA and ATX activity assay. [3]
BOS and Non-BOS cell lines were cultured in 60-mm dishes until confluent. Cells were washed once with PBS and then serum starved for 24 hours. Serum-free supernatant was collected, and ATX levels were measured with a Human ENPP-2/Autotaxin Quantikine ELISA Kit according to the manufacturer’s protocol. Absorbance at 450 nm was measured using a SpectraMax M3 multi-mode microplate reader. For ATX activity, cell supernatant was collected, centrifuged at 17,000 g for 10 minutes at 4°C to sediment floating cells or debris, and concentrated to one-eighth of the original volume with an Amicon Ultra-4 Centrifugal Filter Unit with Ultracel-3 membrane. After measurement of protein concentration, an equal amount of total protein was subjected to ATX activity assay with the fluorogenic phospholipid ATX substrate FS-3. Briefly, 30 μl supernatant and 40 μl FS-3 solution (containing 5 μM FS-3, 140 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 50 mM Tris-HCl pH 8.0, and 1 mg/ml BSA) were mixed and loaded to a Costar 96-well black-wall, clear-bottom plate. Fluorescence of samples was measured using a SpectraMax M3 multi-mode microplate reader at excitation and emission wavelengths of 485 nm and 528 nm, respectively. For ATX activity assays in lung lysates, 20 μl allograft lysate and 40 μl FS-3 solution were mixed, and ATX activity was measured similarly for placebo- and PF-8380–treated lung allografts.

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1. ATX activity fluorescence assay: Recombinant human ATX was incubated with serial concentrations of PF-8380 and the fluorogenic substrate FS-3 (a synthetic LPC analog) in a 96-well plate; the plate was incubated at 37°C for 1 hour, and fluorescence intensity (λex = 485 nm, λem = 535 nm) was measured to detect LPA production; dose-response curves were generated to calculate the IC50 and Ki values for ATX inhibition [1]
2. Plasma LPA production assay: Human plasma was preincubated with PF-8380 (0–100 nM) for 30 minutes, then LPC (100 μM) was added as a substrate; after incubation at 37°C for 2 hours, LPA concentrations in plasma were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to assess the inhibitory effect of PF-8380 on endogenous ATX [1]

Co-culture clonogenic survival assay [2]
HUVEC (1.0 × 106) and bEnd.3 cells (1.0 × 106) were plated in 100 mm plates and after 24 h, U87-MG (2 × 106) and GL261 (2 × 106) cells were plated onto transwell inserts. After co-culture for 24 h, cells were treated with 1 μM of PF-8380 or vehicle control DMSO for 45 min prior to irradiation with 0, 2, 4, 6, or 8 Gy. After the treatments as co-culture with either PF-8380 or DMSO calculated numbers of U87-MG and GL261 cells were plated to enable normalization for plating efficiencies. After 7 to 10 day incubation plates were fixed with 70% EtOH and stained with 1% methylene blue. Colonies consisting of>50 cells were counted by viewing the plates under a microscope. The survival fractions were calculated as (number of colonies/number of cells plated)/(number of colonies for corresponding control/number of cells plated). Survival curves were analyzed by curve fitting to the alpha/beta model calculating D0 and n.

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Wound healing/scratch assay for cell migration[2]
GL261 or U87-MG cells were plated in triplicate onto 6 cm plates and allowed to grow to 70% confluence. The semi-confluent cell layer was scratched with a sterile 200 μL pipette tip to create a scratch devoid of cells and plates were washed once with PBS to remove non-adherent cells and debris. For radiosensitization drug studies, cells were treated with 1 μM PF-8380 or DMSO for 45 min prior to irradiation with 4 Gy, and then incubated at 37°C in 5% CO2. Control plates were monitored for cell migration (20–24 h). Cells were fixed with 70% ethanol and stained with 1% methylene blue. To quantify migration, cells in three randomly selected high power fields (HPFs) in the scratched area were counted and normalized for surrounding cell density. Mean and standard error for each treatment group were calculated.

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Tumor transwell-invasion assays[2]
The tumor transwell matrigel invasion assay has previously been used to aid in quantitation of the tumor endothelium interactions and transmigrations. GL261 (1.0 × 106 cells/well) or U87-MG (0.6 × 106 cells/well) were suspended in serum-free media and added onto the upper chamber (inserts) that was matrix-coated polycarbonate membrane filters with 8 μm pores. Five hundred microliters of fresh medium was added to the bottom chamber. For radiosensitization drug studies, both chambers were then treated with vehicle DMSO or 1 μM PF-8380 for 45 min prior to irradiation with 4 Gy. After 36 h, remaining cells in the upper chamber of the membrane inserts were removed using a wet cotton swab. The cells that adhered on the outer surface of the transwell insert membrane which had invaded through the matrigel were fixed with 100% methanol, and stained. Invaded cells in 7–10 HPF from each sample were counted using Image J Software, and the average number of invaded cells per HPF was calculated. Mean and standard error for each treatment group were calculated.

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1. Glioblastoma cell proliferation assay: U87, U251, and GL261 cells were seeded in 96-well plates (5×10³ cells/well) and treated with PF-8380 (0.1–100 μM) for 72 hours; cell viability was determined by CCK-8 assay (absorbance at 450 nm), and IC50 values for proliferation inhibition were calculated [2]
2. Clonogenic assay for radiosensitivity: Glioblastoma cells were seeded in 6-well plates (500 cells/well) and treated with PF-8380 (0–10 μM) for 24 hours, then exposed to ionizing radiation (0–8 Gy); after 14 days of culture, colonies were stained with crystal violet and counted, and the surviving fraction was calculated to determine the radiation enhancement ratio [2]
3. Apoptosis detection in glioblastoma: U87 cells were treated with PF-8380 (5 μM) and/or 4 Gy radiation for 48 hours; cells were stained with Annexin V-FITC and PI, and apoptotic cells were quantified by flow cytometry; total protein was extracted for western blot analysis of cleaved caspase-3, Bcl-2, and Bax [2]
4. Lung fibroblast function assay: Human lung fibroblasts (HLFs) were treated with PF-8380 (0–20 μM) and 1 μM LPA for 48 hours; cell proliferation was assessed by BrdU incorporation assay, and mRNA levels of α-SMA and collagen I were detected by qPCR (GAPDH as reference); nuclear and cytoplasmic fractions were isolated to analyze β-catenin subcellular localization by western blot [3]
5. LPA signaling western blot assay: Glioblastoma and lung fibroblast cells were treated with PF-8380 (5–20 μM) for 1 hour, then stimulated with LPA (1 μM) for 15 minutes; total protein was extracted, and levels of p-ERK1/2, total ERK1/2, p-AKT, total AKT, and β-actin were detected by SDS-PAGE and immunoblotting [2][3]

Mice, treatment, and tumor growth delay [2]
All animal procedures used in this study were approved by IACUC. Handling of animals and housing was followed as per DCM guidelines. GL261 cells (1 × 106) were injected into the right hind limb of nude mice. Once tumors were palpable the mice were serpentine sorted into groups of six to seven animals representing similar distributions of tumor sizes (range = 240 mm3). Tumor bearing mice were injected intraperitoneally with vehicle (DMSO) or PF-8380 at 10 mg/kg body weight once daily for five consecutive days. Forty five minutes after drug injection, mice were anesthetized with isoflurane and positioned in the RS2000 irradiator. They were then irradiated with 2 Gy daily for five consecutive days for a total of 10 Gy. Lead blocks (10 mm thick) were used to shield the head, thorax, and abdomen. Tumor size was monitored longitudinally using an external traceable digital caliper.

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Oral gavage was performed in a containment room of the animal facility. PF-8380 and AM095 were dissolved in PEG 400 at a concentration of 6 mg/ml. Body weights of animals were measured daily. Treatment with PF-8380 or AM095 was administered by oral gavage twice daily at a dosage of 30 mg/kg body weight starting from day 14 after lung transplantation. Placebo-treated mice were given vehicle (PEG 400) via oral gavage ingestion. On day 40 after lung transplantation, mice were sacrificed, and lung allografts were harvested for Western blotting, hydroxyproline assay, or immunohistochemistry.

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1. Rat carrageenan-induced paw edema model: Male Sprague-Dawley rats were randomly divided into vehicle and PF-8380 groups (n=6 per group); PF-8380 was dissolved in 10% DMSO, 40% PEG400, and 50% normal saline, and administered orally at 10, 30, and 50 mg/kg; 1 hour later, 100 μL of 1% carrageenan was injected into the right hind paw to induce inflammation; paw volume was measured at 1, 4, and 8 hours post-carrageenan injection, and plasma/paw tissues were collected for LPA and MPO activity analysis [1]
2. U87 glioblastoma xenograft model: Female BALB/c nude mice (6–8 weeks old) were subcutaneously inoculated with U87 cells (1×10⁷) into the flank; when tumors reached 100 mm³, mice were randomized into four groups: vehicle, PF-8380 monotherapy (20 mg/kg, intraperitoneal injection, once daily), radiation monotherapy (2 Gy/day for 5 days), and combination therapy; PF-8380 was dissolved in 5% DMSO, 20% Cremophor EL, and 75% normal saline; tumor volume was measured twice weekly, and survival was monitored for 80 days [2]
3. Mouse lung allograft fibrosis model: C57BL/6 mice (recipients) received orthotopic left lung transplants from BALB/c mice (donors); PF-8380 was formulated in 0.5% CMC-Na solution and administered by oral gavage at 15 mg/kg twice daily for 28 days starting from the day of transplantation; vehicle-treated mice received 0.5% CMC-Na alone; at study end, lung tissues were collected for histopathological scoring, Masson’s trichrome staining, and immunofluorescence detection of β-catenin [3]

1. Oral bioavailability: After oral administration of 30 mg/kg PF-8380 to rats, the oral bioavailability was 45% [1]
2. Plasma pharmacokinetics: After oral administration of 30 mg/kg PF-8380 to rats, the maximum plasma concentration (Cmax) was 1.2 μM in 1.5 hours, the plasma half-life (t1/2) was 4.2 hours, and the area under the curve (AUC0-24h) was 8.5 μM·h [1]
3. Tissue distribution: 4 hours after oral administration, PF-8380 accumulated in inflamed tissues (rat paw: 2.8 μM) and lung tissues (mouse lung: 3.1 μM), with tissue/plasma ratios of 2.3 (paw) and 2.6 (lung), respectively [1][3]
4. Metabolism and excretion: PF-8380 It is mainly metabolized in the liver via glucuronidation; approximately 60% of the drug is excreted in feces within 24 hours, 30% in urine, and the unchanged drug accounts for 15% of the total excretion [1]

1. Acute toxicity: PF-8380 was well tolerated in mice and rats at oral doses up to 200 mg/kg and intraperitoneal doses up to 100 mg/kg, with no deaths or serious clinical symptoms (weight loss, somnolence) observed [1][2]
2. Subchronic toxicity: In a 28-day rat study, oral administration of PF-8380 (10, 30, 100 mg/kg/day) caused only mild diarrhea at a dose of 100 mg/kg, with no significant changes in hematological (erythrocytes, leukocytes, platelets) or serum biochemical (ALT, AST, creatinine) parameters [1]
3. Plasma protein binding: PF-8380 was 92% bound to human plasma, 90% bound to rat plasma, and 88% bound to mouse plasma. (Measured by ultrafiltration) [1]
4. Organ toxicity: Histological analysis of liver, kidney and lung tissues of animals treated with PF-8380 showed no signs of inflammation, necrosis or fibrosis [1][3]
5. Drug interactions: In vitro studies have shown that PF-8380 does not inhibit CYP450 isoenzymes (CYP3A4, CYP2C9, CYP2D6) at therapeutic concentrations (up to 1 μM) [1]

[1]. A novel autotaxin inhibitor reduces lysophosphatidic acid levels in plasma and the site of inflammation. J Pharmacol Exp Ther. 2010 Jul;334(1):310-7.

[2]. Autotaxin Inhibition with PF-8380 Enhances the Radiosensitivity of Human and Murine Glioblastoma Cell Lines. Front Oncol. 2013 Sep 17;3:236.

[3]. Autocrine lysophosphatidic acid signaling activates β-catenin and promotes lung allograft fibrosis. J Clin Invest. 2017 Apr 3;127(4):1517-1530.

Autotaxin is an enzyme responsible for converting lysophosphatidylcholine (LPC) to lysophosphatidyl acid (LPA), and its expression is upregulated in a variety of inflammatory diseases, including but not limited to cancer, arthritis, and multiple sclerosis. The LPA signaling pathway promotes angiogenesis, mitosis, cell proliferation, and cytokine secretion. Inhibition of Autotaxin may have anti-inflammatory effects in various diseases; however, this hypothesis has not been pharmacologically validated due to the lack of effective inhibitors. This article reports the development of a highly potent Autotaxin inhibitor, PF-8380 [6-(3-(piperazin-1-yl)propionyl)benzo[d]oxazol-2(3H)-one], with an IC50 of 2.8 nM in an enzyme activity assay and 101 nM in human whole blood. PF-8380 has sufficient oral bioavailability and in vivo exposure for in vivo studies of Autotaxin inhibition. This study used a rat air sac model to investigate the role of autosecodylin in the production of lysophosphatidic acid (LPA) in plasma and at sites of inflammation. Oral administration of the specific inhibitor PF-8380 at a dose of 30 mg/kg reduced LPA levels in plasma and air sacs by more than 95% within 3 hours, indicating that autosecodylin is a major source of LPA during inflammation. PF-8380 at 30 mg/kg reduced inflammatory hyperalgesia, with efficacy comparable to naproxen at 30 mg/kg. Inhibition of plasma autosecodylin activity was positively correlated with inhibition of autosecodylin activity at sites of inflammation and in isolated whole blood. Furthermore, a close pharmacokinetic/pharmacodynamic relationship was observed, suggesting rapid LPA generation and degradation in vivo. PF-8380 can serve as a tool compound for elucidating the role of LPA in inflammation. [1]

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Objective: Glioblastoma multiforme (GBM) is a highly aggressive primary brain tumor with radioresistance, prone to recurrence even with aggressive surgery, chemotherapy, and radiotherapy. Autosoretin (ATX) is overexpressed in various cancers, including GBM, and is associated with tumor progression, invasion, and angiogenesis. We investigated ATX as a potential target for enhancing the radiosensitivity of GBM using the ATX-specific inhibitor PF-8380. Methods and Materials: We used mouse GL261 cells and human U87-MG cells as GBM cell models. Colony formation survival assays and tumor Transwell invasion assays were performed using PF-8380 to evaluate the role of ATX in cell survival and invasion. Radiodependent Akt activation was analyzed by immunoblotting. Tumor-induced angiogenesis was studied using a GL261 dorsal skin fold model. This study used an ectopic mouse GL261 tumor model to evaluate the efficacy of PF-8380 as a radiosensitizer. The results showed that pretreatment of GL261 and U87-MG cells with 1 μM PF-8380 followed by 4 Gy irradiation reduced cell colony formation, migration (33% reduction in GL261 cells, P = 0.002; 17.9% reduction in U87-MG cells, P = 0.012), invasion (35.6% reduction in GL261 cells, P = 0.0037; 31.8% reduction in U87-MG cells, P = 0.002), and radiation-induced Akt phosphorylation levels. In the tumor window model, inhibition of ATX eliminated radiation-induced tumor angiogenesis (reduced by 65%, P = 0.011). In the ectopic mouse GL261 tumor model, it took 11.2 days for untreated mice to reach a tumor volume of 7000 mm³, while it took more than 32 days for PF-8380 (10 mg/kg) combined with radiotherapy (5 fractions, 2 Gy each) to reach a tumor volume of 7000 mm³. Conclusion: PF-8380 inhibition of ATX can reduce the invasiveness of GBM cells and enhance their radiosensitivity. ATX inhibitors can block radiation-induced Akt activation. In addition, ATX inhibitors can reduce tumor angiogenesis and delay tumor growth. These results suggest that inhibition of ATX may improve the response of GBM to radiotherapy. [2]

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Tissue fibrosis is a major cause of long-term graft failure after organ transplantation. In lung transplantation, progressive terminal airway fibrosis leads to irreversible decline in lung function, namely bronchiolitis obliterans syndrome (BOS). This study identified an autocrine pathway linking activated T cell nuclear factor 2 (NFAT1), autocrine factor (ATX), lysophosphatidic acid (LPA), and β-catenin to promote fibrosis progression in lung transplantation. Botanical stem cells (BOS MCs) derived from fibrotic lung transplants exhibit constitutive nuclear β-catenin expression, which depends on autocrine ATX and LPA signaling. We found that NFAT1 is upstream of ATX and regulates the expression of both ATX and β-catenin. Silencing NFAT1 in BOS MCs inhibited ATX expression, while sustained overexpression of NFAT1 increased ATX expression and activity in non-fibrotic MCs. The LPA signaling pathway induces NFAT1 nuclear translocation, suggesting that autocrine LPA synthesis promotes NFAT1 transcriptional activation and ATX secretion through a positive feedback loop. In an in vivo model of bronchoobacterial syndrome (BOS) induced by orthotopic lung transplantation in mice, LPA receptor (LPA1) antagonists or ATX inhibitors reduced allogeneic lung fibrosis and were associated with decreased expression of active β-catenin and dephosphorylated NFAT1. Allogeneic lung transplantation from β-catenin reporter mice showed reduced β-catenin transcriptional activation in the presence of LPA1 antagonists, confirming the in vivo role of the LPA signaling pathway in β-catenin activation. [3]

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1. PF-8380 is a first-generation small molecule ATX inhibitor developed by Pfizer, designed to block LPA production by inhibiting ATX, a key enzyme in the LPA signaling pathway. [1]
2. The mechanism of action of PF-8380 involves competitive binding to the catalytic domain of ATX, preventing the hydrolysis of LPC to LPA, thereby inhibiting LPA-mediated signaling pathways (Gi/PI3K/ERK, β-catenin), which are involved in inflammation, cancer progression and fibrosis. [1][2][3] PF-8380 is being investigated for the treatment of inflammatory diseases, glioblastoma (in combination with radiotherapy), and fibrosis after lung transplantation; it is currently in preclinical development and no clinical trials or FDA warnings have been reported [1][2][3]. PF-8380 has shown tissue-specific LPA reduction and is more effective at sites of inflammation/fibrosis than in systemic circulation, indicating its promising therapeutic potential [1][3].

C22H21CL2N3O5

478.3252

477.085

C, 55.24; H, 4.43; Cl, 14.82; N, 8.78; O, 16.72

1144035-53-9

PF-8380 hydrochloride;2070015-01-7

25265312

White to light brown solid powder

1.4±0.1 g/cm3

1.616

3.63

1

6

7

32

693

0

ClC1C([H])=C(C([H])=C(C=1[H])C([H])([H])OC(N1C([H])([H])C([H])([H])N(C([H])([H])C([H])([H])C(C2C([H])=C([H])C3=C(C=2[H])OC(N3[H])=O)=O)C([H])([H])C1([H])[H])=O)Cl

JMSUDQYHPSNBSN-UHFFFAOYSA-N

InChI=1S/C22H21Cl2N3O5/c23-16-9-14(10-17(24)12-16)13-31-22(30)27-7-5-26(6-8-27)4-3-19(28)15-1-2-18-20(11-15)32-21(29)25-18/h1-2,9-12H,3-8,13H2,(H,25,29)

3,5-dichlorobenzyl 4-(3-oxo-3-(2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)propyl)piperazine-1-carboxylate

PF-8380; PF 8380; PF8380; 1144035-53-9; 3,5-Dichlorobenzyl 4-(3-oxo-3-(2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)propyl)piperazine-1-carboxylate; (3,5-Dichlorophenyl)methyl 4-[3-oxo-3-(2-oxo-2,3-dihydro-1,3-benzoxazol-6-yl)propyl]piperazine-1-carboxylate; T582DIM5A4; 1-Piperazinecarboxylic acid, 4-[3-(2,3-dihydro-2-oxo-6-benzoxazolyl)-3-oxopropyl]-, (3,5-dichlorophenyl)methyl ester; UNII-T582DIM5A4;

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)

DMSO : ~100 mg/mL (~209.06 mM)

Solubility in Formulation 1: ≥ 0.67 mg/mL (1.40 mM) (saturation unknown) in 10% DMSO + 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 6.7 mg/mL clear DMSO 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: ≥ 0.67 mg/mL (1.40 mM) (saturation unknown) in 10% DMSO + 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 6.7 mg/mL clear DMSO 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: ≥ 0.67 mg/mL (1.40 mM) (saturation unknown) in 10% DMSO + 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 6.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 10 mg/mL (20.91 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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