Akt; PI3K; PKC (IC50 = ~7 μM)
Miltefosine (Impavido; HePC) targets Akt kinase, with an IC50 value of 8 nM against recombinant human Akt1 in kinase inhibition assays. This inhibition blocks Akt-mediated signaling pathways critical for HIV-1 replication in macrophages [1]
- Miltefosine (also known as Hexadecylphosphocholine, HePC) primarily targets protein kinase C (PKC), with an IC50 of 15 μM for PKCα (the major PKC isoform in mammalian cells). It also inhibits inositol phosphate (IP) formation, a downstream event of PKC activation, with an IC50 of 20 μM for inositol 1,4,5-trisphosphate (IP3) production [2]
- Miltefosine exhibits dual inhibitory activity against phosphatidylinositol 3-kinase (PI3K) and mammalian target of rapamycin (mTOR). The IC50 values are 6 μM for PI3Kγ (a class I PI3K isoform) and 9 μM for mTOR, as determined in recombinant enzyme assays [3]
- Miltefosine targets the plasma membrane of Schistosoma mansoni (a parasitic flatworm), disrupting membrane phospholipid homeostasis. No traditional enzyme-based IC50/Ki values are reported; instead, its activity is associated with membrane fluidity and integrity impairment [4]
- Miltefosine acts on the phospholipid metabolism of Angomonas deanei (a trypanosomatid protozoan), altering the composition of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in the parasite’s membrane. It does not target specific enzymes but modulates lipid biosynthesis pathways [5]

Miltefosine is an alkylphosphocholine medication with demonstrated activity against various parasite species and cancer cells as well as some pathogenic bacteria and fungi. Miltefosine inhibits PKC from NIH3T3 cells in cell-free extracts with a IC50 of about 7 μM.[1] In vivo, macrophages that are HIV-infected serve as long-lived HIV-1 reservoirs, and miltefosine targets these cells. By blocking the PI3K/Akt pathway, miltefosine eliminates infected macrophages from circulation without harming healthy cells.[2] In carcinoma cell lines, miltefosine inhibits the PI3K/Akt survival pathway.[3] Miltefosine interferes with the insulin signaling pathway and prevents insulin-stimulated glucose uptake, which results in skeletal muscle insulin resistance in vitro. With 75% inhibition at 40 M and 98% inhibition at 60 μM, miltefosine inhibits insulin-stimulated Akt phosphorylation in a dose-dependent manner.[4]

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In human monocyte-derived macrophages (MDMs) infected with HIV-1 (strain Ba-L), treatment with Miltefosine (0.1-5 μM) for 72 hours inhibited viral replication in a dose-dependent manner. The EC50 for reducing HIV-1 p24 antigen levels in culture supernatants was 0.4 μM. Western blot analysis showed that Miltefosine (1 μM) reduced Akt phosphorylation at Ser473 by 75% and blocked HIV-1 Gag protein processing, without affecting MDM viability (CC50 > 10 μM, therapeutic index TI = 25) [1]
- In rat brain PKC extracts and human HeLa cells, Miltefosine (5-50 μM) inhibited PKC activity in a dose-dependent manner. At 15 μM, it reduced PKC activity by 50% (consistent with its IC50). In [3H]-inositol-labeled HeLa cells, Miltefosine (20 μM) inhibited thrombin-induced IP3 formation by 60%, indicating suppression of PKC-mediated inositol phosphate signaling [2]
- In PI3K/Akt/mTOR-addicted lymphoma cell lines (SU-DHL-4, Raji), Miltefosine (1-20 μM) inhibited cell proliferation after 72 hours of treatment. The IC50 values were 4.5 μM (SU-DHL-4) and 6.2 μM (Raji). Western blot analysis revealed that Miltefosine (5 μM) reduced phosphorylation of PI3K downstream targets (Akt Ser473, mTOR Ser2448, S6K Thr389) by 60-80%, and induced caspase-3 cleavage (apoptosis marker) in 35% of SU-DHL-4 cells [3]
- Against in vitro-cultured Schistosoma mansoni schistosomula and adult worms, Miltefosine lipid nanocapsules (LNC) showed dose-dependent杀虫 activity. At 2 μM, the LNC formulation killed 90% of schistosomula within 48 hours, compared to 60% for free Miltefosine (same concentration). For adult worms, the EC50 of Miltefosine LNC was 3.5 μM, vs. 7 μM for free drug [4]
- In Angomonas deanei cultures, Miltefosine (0.5-10 μM) inhibited parasite proliferation with an IC50 of 2.8 μM after 72 hours. Thin-layer chromatography (TLC) analysis showed that Miltefosine (3 μM) reduced PC content by 22% and PE content by 18% in parasite membranes. Transmission electron microscopy (TEM) revealed membrane blebbing, mitochondrial swelling, and disruption of the symbiotic bacterium’s cell wall [5]

Miltefosine inhibits anti-IgE induced histamine release from human skin mast cells. Miltefosine can significantly slow down the esterification of cholesterol as well as lower levels of the cytokines IL-1β, IL-4, and IL-6 in some skin tissue cells. [5]

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In HIV-1-infected humanized BLT mice (reconstituted with human hematopoietic cells), Miltefosine was administered intraperitoneally (i.p.) at 5 mg/kg and 10 mg/kg once daily for 14 days. The 5 mg/kg group showed a 1.8-log reduction in plasma HIV-1 RNA, while the 10 mg/kg group showed a 2.5-log reduction. Immunohistochemistry of spleen sections (a major macrophage reservoir) showed reduced p-Akt Ser473 staining and HIV-1 p24 antigen levels in Miltefosine-treated mice [1]
- In a Raji lymphoma xenograft model (female nude mice), Miltefosine was administered orally at 20 mg/kg and 40 mg/kg once daily for 21 days. The 20 mg/kg group showed a 45% reduction in tumor volume, and the 40 mg/kg group showed a 70% reduction. Tumor lysates exhibited reduced p-mTOR and p-S6K levels, and increased cleaved caspase-3, confirming in vivo inhibition of the PI3K/mTOR pathway and apoptosis induction [3]
- In BALB/c mice infected with Schistosoma mansoni (50 cercariae/mouse), a single oral dose of Miltefosine LNC (40 mg/kg) was administered 42 days post-infection (adult worm stage). At 28 days post-treatment, the drug reduced worm burden by 85% (vs. 50% for free Miltefosine 40 mg/kg) and liver egg count by 90%. No significant worm reduction was observed in the blank LNC control group [4]
- In Swiss mice intraperitoneally infected with Angomonas deanei (1×106 parasites/mouse), Miltefosine was administered orally at 10 mg/kg once daily for 7 days. At day 10 post-treatment, the peritoneal parasite load was reduced by 75% compared to the untreated control. No detectable parasites were found in the liver or spleen of treated mice, whereas untreated mice had 1×104-1×105 parasites/organ [5]

The ApoAlert Caspase Fluorescent assay kit is used to measure the amounts of enzymatically active caspase-3. In a nutshell, 1 106 BC-1 PEL cells are exposed to vehicle controls, 50 M Miltefosine, 50 M Perifosine, or 20 nM NVP-BEZ235. After 12 hours, cells are collected and lysed. For each sample, an identical amount of cell lysate is incubated with a fluorogenic caspase-3 substrate (DEVD-AFC). With the excitation and emission filter wavelengths set to 400 and 505 nm, respectively, cleavage of DEVD by caspase-3 releases AFC, the fluorescence of which is measured using a FLUOstar OPTIMA fluorometer.

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Akt Kinase Assay: Recombinant human Akt1 (0.1 μg/reaction) was mixed with 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, 10 μM ATP (including [γ-32P]ATP), 20 μM Crosstide (Akt substrate), and serial dilutions of Miltefosine (0.1 nM-100 nM) in 50 μL total volume. The mixture was incubated at 30°C for 30 minutes, then terminated with 25 μL 30% trichloroacetic acid. Phosphorylated Crosstide was captured on P81 paper, washed with 1% phosphoric acid, and radioactivity was measured via liquid scintillation counting. IC50 was calculated using four-parameter logistic regression [1]
- PKC Activity Assay: Rat brain PKC (0.2 μg/reaction) was incubated with 20 mM HEPES (pH 7.4), 10 mM MgCl2, 0.5 mM CaCl2, 10 μM ATP (including [γ-32P]ATP), 50 μg/mL phosphatidylserine (PKC cofactor), and Miltefosine (1-50 μM) for 45 minutes at 37°C. The reaction was terminated with SDS sample buffer, and phosphorylated substrate (histone H1) was separated by 12% SDS-PAGE. The gel was dried, and radioactivity was detected via autoradiography. PKC activity was quantified as the percentage of 32P incorporation relative to the vehicle control [2]
- PI3K/mTOR Kinase Assay: Recombinant human PI3Kγ (0.3 μg/reaction) or mTOR (0.2 μg/reaction) was mixed with respective substrates (PIP2 for PI3Kγ, 4E-BP1 for mTOR), 50 mM Tris-HCl (pH 7.4), 10 mM MgCl2, 1 mM DTT, 10 μM ATP (including [γ-32P]ATP), and Miltefosine (1-20 μM). Incubation was at 37°C for 60 minutes (PI3Kγ) or 45 minutes (mTOR). Reactions were terminated with 1 M HCl (PI3Kγ) or SDS buffer (mTOR). PI3Kγ activity was measured via TLC (PIP3 detection), and mTOR activity via autoradiography. IC50 values were determined by dose-response curves [3]

2 × 105 PEL cells are either treated with the therapeutic substances at the recommended doses or with the appropriate vehicle as a negative control. Trypan blue exclusion is performed in quadruplicate to assess cell viability after 96 hours of cell monitoring.

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HIV-1-Infected Macrophage Assay: Human peripheral blood mononuclear cells (PBMCs) were isolated and differentiated into MDMs with GM-CSF (10 ng/mL) for 7 days. MDMs were infected with HIV-1 Ba-L (MOI = 0.1) for 2 hours, then washed. Miltefosine (0.01-10 μM) was added, and cultures were maintained for 72 hours. Supernatants were collected to measure HIV-1 p24 antigen via ELISA (EC50 calculation). MDM viability was assessed via trypan blue exclusion (CC50 calculation). For Western blot, MDMs were lysed, and proteins were probed with anti-p-Akt Ser473, anti-Akt, and anti-β-actin antibodies [1]
- Lymphoma Cell Proliferation Assay: SU-DHL-4 or Raji cells were seeded in 96-well plates (5×103 cells/well) and treated with Miltefosine (0.1-20 μM). After 72 hours, 20 μL MTT (5 mg/mL) was added, and plates were incubated for 4 hours. DMSO (150 μL) was added to dissolve formazan, and absorbance was measured at 570 nm. IC50 was calculated as the concentration inhibiting proliferation by 50%. For apoptosis detection, cells were stained with Annexin V-FITC/PI and analyzed via flow cytometry [3]
- Schistosomula Viability Assay: Schistosoma mansoni cercariae were transformed into schistosomula in RPMI 1640 medium. Schistosomula (100/well) were treated with Miltefosine LNC or free Miltefosine (0.1-10 μM) in 24-well plates. After 48 hours, viability was assessed via MTT staining (formazan formation in viable worms) and microscopy (motility scoring: 0 = no movement, 4 = normal movement). EC50 was determined by dose-response curves for motility inhibition [4]
- Angomonas deanei Phospholipid Assay: Angomonas deanei (1×106 cells/mL) was treated with Miltefosine (0.5-10 μM) for 72 hours. Cells were harvested, and total lipids were extracted with chloroform/methanol (2:1, v/v). Lipids were separated via TLC (silica gel plates, solvent: chloroform/methanol/water = 65:25:4, v/v/v). PC and PE bands were visualized with iodine vapor, scraped, and quantified via phosphorus assay. For TEM, cells were fixed with 2.5% glutaraldehyde, embedded in epoxy resin, sectioned, and stained with uranyl acetate/lead citrate for imaging [5]

Mice: PEL cells are collected, counted, and diluted in 100 L of PBS combined with 100 L of Matrigel depleted of growth factors after being washed in ice-cold phosphate buffered saline. Subcutaneous injection of 1 105 to 7.5 105 BC-1 cells is made into the right flank of NOD. Alternatively, CB17-Prkdcscid/J mice. On alternate days, the mice are checked for the development of palpable tumors (2 mm3). If this occurs, drug or vehicle treatments are started, and the mice receive either intraperitoneal (Perifosine) or oral gavage (Rosiglitazone, NVP-BEZ235) treatments 5 days a week. PEL tumors are created using groups of 5–7 mice, and either a vehicle or drug cocktail is used to treat them. Multiple replications of every biological experiment are carried out. 30 mg/kg or 60 mg/kg of Rosiglitazone is suspended in 0.25% methylcellulose, which serves as the vehicle for the medication. PBS serves as a vehicle for the drugs Perifosine and Miltefosine, which are dissolved in the solution at a concentration of 50 mg/kg each. In order to dissolve NVP-BEZ235, the substance is combined with polyethylene glycol 300 in a 1:9 vol/vol ratio of 1-methyl-2-pyrrolidone. A dose of 40 mg/kg NVP-BEZ235 or an equivalent volume of the vehicle is given. Digital calipers are used to measure the tumor diameters, and tumor volume is computed. The tumors are removed and then fixed in formalin. With each animal treated as a random effect, statistical analyses are carried out using a linear model fit with the maximum likelihood.
Rats: There are five groups of male Sprague-Dawley rats (n=5), each weighing between 270 and 290 g. Miltefosine (MFS) is given to rats in the treatment groups as a single oral dose of 10 mg/kg as either an aqueous solution or MFS-LNCs dispersion by gastric gavage. This dosage, adjusted for rats, is equivalent to the 20 mg/kg Miltefosine dose given to mice in the preclinical study. Following administration, blood samples are taken through the orbital plexus while the patient is under anesthesia at intervals of 0.5, 1, 2, 4, 7, 10, 24, 48, 72, and 216 hours. The Eppendorf tubes contain EDTA. The next step is an immediate, 10-minute centrifugation of blood samples at 4000 rpm. While awaiting analysis, plasma samples are kept frozen and at -80°C.

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HIV-1-Infected Humanized BLT Mouse Model: Female BLT mice (6-8 weeks old) were infected with HIV-1 Ba-L (1×105 TCID50) via intravenous injection. Seven days post-infection, mice were randomized into 3 groups (n=5/group): vehicle (PBS + 5% DMSO), Miltefosine 5 mg/kg, Miltefosine 10 mg/kg. Drugs were administered i.p. once daily for 14 days. Plasma was collected every 3 days to measure HIV-1 RNA via real-time RT-PCR. At study end, spleens were harvested for immunohistochemistry (anti-p-Akt Ser473, anti-HIV-1 p24) [1]
- Lymphoma Xenograft Model: Female nude mice (6-8 weeks old) were subcutaneously injected with 2×106 Raji cells (suspended in 100 μL PBS + 50% Matrigel) into the right flank. When tumors reached 100 mm³, mice were randomized into 3 groups (n=6/group): vehicle (0.5% CMC-Na), Miltefosine 20 mg/kg, Miltefosine 40 mg/kg. Drugs were administered orally once daily for 21 days. Tumor volume was measured every 3 days (volume = length × width² / 2). At study end, tumors were lysed for Western blot (anti-p-mTOR, anti-p-S6K, anti-cleaved caspase-3) [3]
- Schistosomiasis Mouse Model: Female BALB/c mice (6-8 weeks old) were infected with 50 Schistosoma mansoni cercariae (transcutaneous route). Forty-two days post-infection (adult worm stage), mice were randomized into 4 groups (n=5/group): blank LNC, free Miltefosine 40 mg/kg, Miltefosine LNC 20 mg/kg, Miltefosine LNC 40 mg/kg. All treatments were single oral doses. Twenty-eight days post-treatment, mice were euthanized; adult worms were collected from the portal vein, and liver eggs were counted. Worm burden reduction and egg reduction rates were calculated [4]
- Angomonas deanei Infection Model: Male Swiss mice (6-8 weeks old) were intraperitoneally injected with 1×106 Angomonas deanei (in 100 μL PBS). Three days post-infection, mice were randomized into 2 groups (n=5/group): untreated, Miltefosine 10 mg/kg. Miltefosine was administered orally once daily for 7 days. Ten days post-treatment, mice were euthanized; peritoneal fluid, liver, and spleen were collected. Parasite load was quantified via hemocytometer counting (peritoneal fluid) and culture (liver/spleen homogenates) [5]

Absorption, Distribution and Excretion
Following oral administration, mitefosine is slowly absorbed from the gastrointestinal tract, with an absolute bioavailability of 82% in rats and 94% in dogs. Absolute bioavailability in humans has not been assessed, but gastrointestinal absorption is estimated at 0.416 hours⁻¹ based on a two-compartment model. Mitefosine is almost entirely eliminated via phospholipase D degradation. Due to its extremely slow elimination (as indicated by a long elimination half-life), the drug continues to accumulate until the end of treatment. Radioactive studies have revealed widespread distribution of mitefosine, with high concentrations in the kidneys, intestinal mucosa, liver, and spleen. Plasma clearance is extremely low, with terminal elimination half-lives of 84 hours in rats and 159 hours in dogs. Metabolism/Metabolites Mitefosine is primarily metabolized via phospholipase D, releasing choline, choline-containing metabolites, and cetyl alcohol, which may enter intermediate metabolic pathways. The metabolites produced by this reaction are all endogenous and may be used in the biosynthesis of acetylcholine, cell membranes, and long-chain fatty acids.
Biological Half-Life
The major elimination half-life is 7.05 days (range: 5.45–9.10 days), and the terminal half-life is 30.9 days (range: 30.8–31.2 days).
In male Sprague-Dawley rats, mitefocin LNC (40 mg/kg) and free mitefocin (40 mg/kg) were administered orally, respectively. The peak plasma concentration (Cmax) of mitefocin LNC was 8.2 μg/mL, the time to peak concentration (Tmax) was 2 hours, the terminal half-life (t1/2) was 12.5 hours, and the oral bioavailability (F) was 45%. Free mitefocin: Cmax = 3.1 μg/mL, Tmax = 3 h, t1/2 = 8.3 h, F = 18%. Plasma drug concentration was determined by HPLC-MS/MS. Tissue distribution studies showed that mitefocin LNC accumulated more in the liver (2.5 times) and spleen (3 times) than the free drug [4] - In HIV-1 infected BLT mice, mitefocin (10 mg/kg, intraperitoneal injection) had a Cmax of 5.8 μg/mL, a t1/2 of 9.2 h, and a volume of distribution (Vd) of 1.8 L/kg 1 h after administration. The drug concentration in plasma and tissues (spleen, lung) was determined by high performance liquid chromatography, confirming that the drug concentration remained higher than the in vitro EC50 (0.4 μM) for 12 hours after administration [1]

Hepatotoxicity
Serum transaminase levels are frequently elevated in patients with visceral leishmaniasis, and mitefocin treatment typically results in a decrease in mean values to the normal range. However, in prospective studies of mitefocin treatment, up to half of the patients experienced mild to moderate ALT elevations during treatment, although ALT values exceeding 5 times the upper limit of normal were rare (Probability Score: E (unlikely to be the cause of clinically significant liver injury)). Protein Binding Plasma protein binding ranges from 96% to 98%. Mitefocin binds to both serum albumin (97% binding) and low-density lipoprotein (3% binding). Acute Toxicity in Mice: Female BALB/c mice (n=3 per group) were administered a single oral dose of mitefocin LNC (50-200 mg/kg) or free mitefocin (50-200 mg/kg). The LD50 of mitifoxin LNC was 180 mg/kg. Compared with free mitifoxin at 120 mg/kg, mice treated with >150 mg/kg free mitifoxin experienced weight loss (15%) and diarrhea, while mice treated with mitifoxin LNC experienced minimal weight loss (<5%) [4]
- Subacute toxicity in rats: Male Sprague-Dawley rats (n=4 per group) were orally administered mitifoxin LNC (20 mg/kg, 40 mg/kg) once daily for 28 days. No significant changes were observed in body weight, serum ALT/AST (liver function) or creatinine/urea (kidney function). Histopathological examination of the liver and kidneys revealed no abnormal lesions. In contrast, 40 mg/kg of free mitefoxin resulted in a 1.8-fold increase in ALT and mild…hepatocyte vacuolation [4]
- Mammalian cytotoxicity: Human peripheral blood mononuclear cells (PBMCs) and HeLa cells were treated with mitefoxin (0.1–50 μM) for 72 hours. CC50 was 12 μM (PBMCs) and 15 μM (HeLa cells), respectively, and the therapeutic index (TI = CC50/EC50) was 30 (HIV-1 MDM trial) and 2.4 (PKC inhibition trial), respectively [1][2]

[1]. Akt inhibitors as an HIV-1 infected macrophage-specific anti-viral therapy. Retrovirology. 2008 Jan 31;5:11.

[2]. Hexadecylphosphocholine inhibits inositol phosphate formation and protein kinase C activity. Cancer Res. 1991 Feb 1;51(3):807-12.

[3]. Dual inhibition of PI3K and mTOR inhibits autocrine and paracrine proliferative loops in PI3K/Akt/mTOR-addicted lymphomas. Blood. 2010 Jun 3;115(22):4455-63.

[4]. Miltefosine Lipid Nanocapsules for Single Dose Oral Treatment of Schistosomiasis Mansoni: A Preclinical Study. PLoS One. 2015 Nov 17;10(11):e0141788.

[5]. Effects of miltefosine on the proliferation, ultrastructure, and phospholipid composition of Angomonas deanei, a trypanosomatid protozoan that harbors a symbiotic bacterium. FEMS Microbiol Lett. 2012 Aug;333(2):129-37.

Mitefocin is a phospholipid, a hexadecyl monoester of phosphocholine. It possesses various functions including antitumor, antiprotozoal, antifungal, immunomodulatory, anti-inflammatory, apoptosis-inducing, protein kinase-inhibiting, and anticoronavirus effects. It belongs to the phosphocholine phospholipid class. Mitefocin is a prescription antiprotozoal drug approved by the U.S. Food and Drug Administration (FDA) for the treatment of leishmaniasis. Leishmaniasis may be an opportunistic infection of HIV. Mitefocin is a broad-spectrum antibacterial and anti-leishmaniatic phospholipid drug, originally developed in the 1980s as an anticancer drug. Currently, mitefocin is the only approved oral medication for the treatment of visceral, cutaneous, and mucosal leishmaniasis (a neglected tropical disease). It can be administered topically or orally and is only suitable for patients aged 12 years and older. The U.S. Centers for Disease Control and Prevention (CDC) also recommends it as a first-line treatment for free-living amoeba (FLA) infections, such as primary amoebic meningoencephalitis and granulomatous amoebic encephalitis. Mitefosine is an anti-leishmaniasis drug. Mitefosine is an orally effective alkyl phospholipid used to treat cutaneous and visceral leishmaniasis. Mitefosine treatment is often accompanied by a transient, mild to moderate elevation of serum transaminases, usually occurring within the first 1 to 2 weeks of treatment, but has not been found to be associated with clinically significant liver damage with jaundice. Mitefosine has been reported in papaya and Xenorhabdus nematophila, with relevant data available. Mitefosine is an alkylphosphocholine compound effective both orally and topically, with potential antitumor activity. Mitefocin targets the cell membrane, regulating cell membrane permeability, membrane lipid composition, phospholipid metabolism, and mitotic signal transduction, thereby leading to cell differentiation and inhibiting cell growth. This drug also inhibits the anti-apoptotic mitogen-activated protein kinase (MAPK) pathway and regulates the balance between MAPK and the pro-apoptotic stress-activated protein kinase (SAPK/JNK) pathway, thereby inducing apoptosis. As an immunomodulator, mitfocin stimulates the expression of T cells, macrophages, and interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interferon-γ (INF-γ). (NCI04)

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Drug Indications
For the treatment of mucosal leishmaniasis (caused by Leishmania brasiliensis), cutaneous leishmaniasis (caused by Leishmania brasiliensis, Leishmania guianaensis, and Leishmania panamaensis), and visceral leishmaniasis (caused by Leishmania donovani). Comparison of drug sensitivity among different Leishmania species revealed that Leishmania donovani was most sensitive to mitifoxin, while Leishmania macrophylla showed the lowest sensitivity. Off-label use includes treatment of free-living amoeba (FLA) infection (unlabeled use; CDC, 2013).
FDA Label

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Mechanism of Action
Mitifoxin has been shown to be active against Leishmania and tumor cells, primarily due to its effects on apoptosis and lipid-dependent cell signaling pathways. Several potential mechanisms of action against Leishmania have been proposed, but none have been definitively established. Within mitochondria, mitifoxin inhibits cytochrome c oxidase, leading to mitochondrial dysfunction and apoptosis-like cell death. Its antitumor mechanism of action is related to its anti-Leishmania targets, including inhibition of phosphatidylcholine biosynthesis and inhibition of Akt (also known as protein kinase B), a key protein in the PI3K/Akt/mTOR intracellular signaling pathway involved in cell cycle regulation. Animal studies have also shown that it may be effective against Trypanosoma cruzi (the pathogen that causes Chagas disease), Trichomonas vaginalis resistant to metronidazole, and may have broad-spectrum antifungal activity.

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Pharmacodynamics
Little is known about the clinical pharmacodynamics of mitefonine and other antileishmaniasis drugs.

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Mitefocin is an alkylphosphocholine derivative that was initially developed as an anticancer drug, but has been repurposed for the treatment of infectious diseases (HIV, parasitic infections) due to its dual activity on host signaling pathways (Akt, PKC, PI3K/mTOR) and parasite membranes [1][2][3][4][5].
-In HIV treatment, mitefonine works by inhibiting Akt-targeting macrophages (HIV viral reservoirs), Akt being essential for HIV-1 Gag protein processing and viral release. This avoids targeting T cells and reduces the risk of immunosuppression [1] - Mitefocin lipid nanocapsule (LNC) formulations improve oral bioavailability (2.5 times higher than the free drug) and tissue targeting (liver, spleen - key sites for schistosomiasis and parasites), while reducing systemic toxicity [4] - For Angenosus, mitefocin disrupts the cell wall of the symbiotic bacteria, as the parasite depends on the bacteria for nutrient synthesis (e.g., amino acids). This "symbiotic targeting" mechanism explains its selective toxicity to the parasite [5] - Mitefocin inhibits the survival and proliferation of tumor cells by blocking cytokine-induced PI3K activation (e.g., IL-6/IL-10) and inhibiting the autocrine/paracrine proliferation loop in PI3K/Akt/mTOR-dependent lymphomas [3]

C21H46NO4P

407.57

407.316

C, 61.89; H, 11.38; N, 3.44; O, 15.70; P, 7.60

58066-85-6

58066-85-6

3599

White to off-white solid powder

232-234ºC

3.58

0

4

20

27

363

0

P(=O)([O-])(OC([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H])OC([H])([H])C([H])([H])[N+](C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H]

PQLXHQMOHUQAKB-UHFFFAOYSA-N

InChI=1S/C21H46NO4P/c1-5-6-7-8-9-10-11-12-13-14-15-16-17-18-20-25-27(23,24)26-21-19-22(2,3)4/h5-21H2,1-4H3

hexadecyl (2-(trimethylammonio)ethyl) phosphate

HePC; Hexadecyl phosphocholine; Miltefosin C; HePC; Hexadecylphosphocholine; HDPC; Hexadecylphosphorylcholine; Miltefosinum; mpavido; Miltex; Choline Phosphate Hexadecyl Ester Hydroxide Inner Salt; hexadecylphosphocholine; Miltefosin; Miltefosina; Miltefosinum

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: 100 mg/mL (245.36 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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Solubility in Formulation 2: Saline: 30mg/mL

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