G-CSF; IL-6; IL-8; TNFα; Neurokinin-1 receptor( IC50 = 0.1 nM)
Aprepitant decreases metabolic activity with an estimated IC50 value of 20 µM. Aprepitant causes G1 cell cycle arrest and inhibition of cell growth. In Nalm-6 cells, apropitant strongly induces apoptosis, which is mediated by activating caspase-3. Pro-apoptotic p53 target gene expression and p53 accumulation are induced by aprepitant (20 µM)[2]. In a dose-dependent manner, aprepitant (1, 5, 10 µM) inhibits HIV infection in MDM from HIV-negative individuals who are depressed and those who are not. Apreciant's IC50 value is approximately 5 μM, while its IC90 value is 10 μM[4].

Aprepitant decreases metabolic activity with an estimated IC50 value of 20 µM. Aprepitant causes G1 cell cycle arrest and inhibition of cell growth. In Nalm-6 cells, apropitant strongly induces apoptosis, which is mediated by activating caspase-3. Pro-apoptotic p53 target gene expression and p53 accumulation are induced by aprepitant (20 µM)[2]. In a dose-dependent manner, aprepitant (1, 5, 10 µM) inhibits HIV infection in MDM from HIV-negative individuals who are depressed and those who are not. Apreciant's IC50 value is approximately 5 μM, while its IC90 value is 10 μM[4].

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Aprepitant inhibits HIV-1 Bal infection of human monocyte-derived macrophages (MDM) ex vivo in a dose-dependent manner, with an ED50 of ~5 µM. A concentration of 10 µM is approximately equivalent to the IC90. Depression status, sex, age, or race had no effect on this inhibition. Assay of aprepitant concentrations in culture media showed little or no degradation during week-long infectivity experiments.
Incubation of peripheral blood mononuclear cells (PBMC) from healthy donors with Substance P (SP) upregulated production of pro-inflammatory cytokines and chemokines (G-CSF, IL-6, IL-8, TNFα, MIP-1α, and MCP-1). Aprepitant (10 µM) blocked this SP-induced upregulation. DMSO vehicle had no effect.
Incubation of PBMC with SP also resulted in increased PD-1 expression on CD4+ T-cells, an effect blocked by co-incubation with aprepitant.
Exposure of human monocytes to SP resulted in rapid, time- and dose-dependent release of soluble CD163 (sCD163), which was inhibited by pre-incubation with aprepitant.

Aprepitant inhibits the upregulation of NK-1R expression brought on by B. burgdorferi infection of NHP in vivo. B. burgdorferi-induced elevations in CCL2 protein levels in NHP CSF are inhibited by aprepitant treatment. Aprepitant treatment inhibits B. burgdorferi-induced increases in the mRNA expression of CCL2 and CXCL13 in the NHPs' dorsal root ganglia as well as CCL2, CXCL13, IL-17A, and IL-6 in the NHPs' spinal cord. The reductions in astrocyte activity/numbers caused by B. burgdorferi infection are lessened by aparepitant treatment[1]. In mice, the locomotor activation and CPP expression induced by AMPH and cocaine are significantly reduced by aparepitant (10 mg/kg, i.p.). Significant CPP, conditioned place aversion, locomotor activation, or suppression are not induced by appetizing substances[3]. Comparing treated with aparepitant (125 mg/day, p.o.), the levels of viral RNA in plasma are reduced by 1 log[4].

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In a rhesus macaque model of Lyme neuroborreliosis, oral administration of aprepitant (average dose 28 ± 6 mg/kg/day) significantly attenuated B. burgdorferi-induced neuroinflammatory responses.
Aprepitant treatment prevented infection-associated increases in NK-1R mRNA and protein expression in the brain cortex at 2 weeks post-infection.
Aprepitant treatment significantly attenuated B. burgdorferi-induced elevations of CCL2 protein levels in the cerebrospinal fluid (CSF) at 2 weeks post-infection in one experimental series.
Aprepitant treatment significantly attenuated infection-associated increases in mRNA expression of inflammatory mediators in specific central nervous system (CNS) tissues: it reduced CXCL13 and CCL2 mRNA expression in dorsal root ganglia (DRG) at 2 and 4 weeks post-infection; it reduced CXCL13, CCL2, and IL-17A mRNA expression in cervical spinal cord and dura mater at 2 weeks post-infection; and it reduced IL-6 mRNA expression in cervical spinal cord at 4 weeks post-infection.
Aprepitant treatment attenuated infection-induced reductions in the expression of the astrocyte marker glial fibrillary acidic protein (GFAP) in the frontal cortex at 2 and 4 weeks post-infection, as determined by immunofluorescence.
No adverse events attributable to aprepitant treatment were recorded in any animal during the study. [1]

Appetitant's inhibitory effect on Nalm-6 cells' metabolic activity is measured by the viable cells' uptake of thiazolyl blue tetrazolium bromide (MTT). At a density of 5000 cells per well, cells are plated onto 96-well plates. Following 24–36–48 hours of aprepitant treatment at 5, 10, 15, 20, and 30 µM, the cells are then incubated for 3 hours at 37°C with 100 μL of MTT (0.5 mg/mL). The control group consists of untreated cells. An ELISA reader is used to measure the optical densitometry at a wavelength of 578 nm after solubilizing the precipitated formazan with 100 μL of DMSO.

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HIV Infectivity in MDM: Monocytes were isolated from human blood samples and differentiated into macrophages (MDM) by culture for 7 days. MDMs were pre-incubated with aprepitant (1, 5, 10 µM) or vehicle (0.001% DMSO) for 2 hours before infection with HIV-1 Bal. After overnight incubation, unbound virus was washed away. Culture medium containing aprepitant was replaced twice weekly. At day 7 post-infection, cellular RNA was extracted and HIV gag mRNA expression was quantified using real-time RT-PCR.
Cytokine Production in PBMC: Freshly isolated human PBMCs were cultured overnight. Cultures were then treated with Substance P (10 µM), aprepitant (10 µM), both, or vehicle (DMSO). Supernatants were collected 24 hours later, and cytokine concentrations were measured using a multiplex magnetic bead panel assay.
PD-1 Expression in PBMC: PBMCs were treated with Substance P (10 µM) and/or aprepitant (10 µM). PD-1 expression on CD3+CD4+ cells was monitored daily for up to 8 days using flow cytometry.
sCD163 Release from Monocytes: Freshly isolated human monocytes were treated with varying doses of Substance P (with or without aprepitant pre-incubation). Supernatants were collected at different time points, and soluble CD163 levels were measured by ELISA.

Fifteen rhesus macaques undergo anesthesia and are intrathecally injected with 1×10⁸ live spirochetes into the cisterna magna. The remaining five macaques remain uninfected and are given 1 mL of RPMI 1640 medium after a corresponding amount of CSF is removed. A positive culture from at least a necropsy tissue sample indicates the establishment of an in vivo B. burgdorferi infection. The first group of animals, which were divided into two control groups (one receiving aprepitant treatment), two infected groups receiving no treatment, and two infected groups receiving aprepitant treatment, were all subjected to a two-week study. The second group of animals, which consists of five infected and untreated animals, four infected animals treated with aprepitant, and three control animals, one of which is treated with aprepitant, are all studied for four weeks. Apepitant is given to animals orally twice a day at an average dose of 28 ± 6 mg/kg, and medication treatments begin two days prior to inoculation. These dosages align with typical veterinary regimens for the selected medications in NHP, and the 4-week study period avoids the development of neural pathology, which happens eight weeks after B. burgdorferi infection.

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A rhesus macaque (Macaca mulatta) model of Lyme neuroborreliosis was used.
Animals (2.5 to 5.5 years old) were anesthetized and inoculated intrathecally into the cisterna magna with 1 × 10^8 live B. burgdorferi spirochetes in RPMI 1640 medium.
Aprepitant was administered orally (p.o.) daily at an average dose of 28 ± 6 mg/kg/day.
Drug treatment was initiated 2 days prior to bacterial inoculation and continued for either 2 or 4 weeks until euthanasia.
Control groups included uninfected animals (some treated with aprepitant), infected but untreated animals, and infected animals treated with aprepitant.
At necropsy, tissues were collected including brain cortex, dorsal root ganglia, spinal cord, and cerebrospinal fluid for subsequent analysis (RT-PCR, immunoblot, multiplex ELISA, immunofluorescence). [1]

Absorption, Distribution and Excretion
The mean absolute oral bioavailability of aprepitant is approximately 60% to 65%. Aprepitant is primarily eliminated through metabolism; it is not excreted by the kidneys. Aprepitant is secreted into rat milk. It is unknown whether this drug is secreted into human milk. Apparent plasma clearance = 62-90 mL/min
hr Metabolism/Metabolites Aprepitant is primarily metabolized via CYP3A4, with minor metabolism via CYP1A2 and CYP2C19. Approximately seven aprepitant metabolites have been identified in human plasma, all of which retain weak pharmacological activity.
The known human metabolites of aprepitant include 5-oxo-1,4-dihydro-1,2,4-triazol-3-carboxaldehyde, 1-[3,5-bis(trifluoromethyl)phenyl]acetophenone, 5-{[(2S,3S)-3-(4-fluorophenyl)-2-hydroxymorpholin-4-yl]methyl}-2,4-dihydro-1,2,4-triazol-3-one, and (2R,3S)-2-((R)-1-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(4-fluorophenyl)morpholine.
Biological half-life
9–13 hours.
Aprepitant is a centrally acting NK-1 receptor antagonist that crosses the blood-brain barrier. It crosses the blood-brain barrier after oral administration.
Human positron emission tomography studies support this view, showing that it can occupy brain NK-1 receptors in a dose- and plasma-concentration-dependent manner. [1]

Hepatotoxicity
In a pre-registration clinical trial of aprepitant, 6% of patients receiving the treatment experienced elevated serum transaminases, compared to 4.3% in a control group receiving cancer chemotherapy. These elevations were transient, mild to moderate, and without symptoms or jaundice. No convincing cases of clinically significant liver injury caused by aprepitant or fosaprepitant have been published in the literature, and therefore, even if they do exist, serious liver injury from the use of these two drugs is extremely rare.
Probability score: E (unlikely to be the cause of clinically significant liver injury).

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Protein binding
Protein binding has been reported to be >95%.

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In this specific study involving rhesus monkeys, no adverse events attributable to the treatment were recorded in any animals treated with aprepitant.
This study cites other studies (in humans and mice) that have not found any safety issues with the use of aprepitant. [1]

[1]. Aprepitant limits in vivo neuroinflammatory responses in a rhesus model of Lyme neuroborreliosis. J Neuroinflammation. 2017 Feb 15;14(1):37.

[2]. Inhibition of tachykinin NK1 receptor using aprepitant induces apoptotic cell death and G1 arrest through Akt/p53 axis in pre-B acute lymphoblastic leukemia cells. Eur J Pharmacol. 2016 Nov 15;791:274-283.

[3]. Differential effects of aprepitant, a clinically used neurokinin-1 receptor antagonist on the expression of conditioned psychostimulant versus opioid reward. Psychopharmacology (Berl). 2017 Feb;234(4):695-705.

[4]. Pharmacologic rationale for the NK1R antagonist, aprepitant as adjunctive therapy in HIV. J Transl Med. 2016 May 26;14(1):148.

Pharmacodynamics
Aprepitant is an antiemetic, belonging to the substance P/neurokine 1 (NK1) receptor antagonist class. Used in combination with other antiemetics, it is indicated for the prevention of acute and delayed nausea and vomiting (NAV) induced by initial and repeated cycles of highly emetogenic chemotherapy in cancers. Aprepitant is a selective, high-affinity antagonist of the human substance P/neurokine 1 (NK1) receptor. Aprepitant has little affinity for serotonin (5-HT3), dopamine, and corticosteroid receptors, which are the targets of existing chemotherapy-induced nausea and vomiting (CINV) therapies. Aprepitant is a specific non-peptide neurokinin-1 receptor (NK-1R) antagonist. It is a drug approved by the U.S. Food and Drug Administration (FDA) and is currently used clinically (along with its prodrug fosaprepitant) as an antiemetic following chemotherapy.
This study explores the potential use of aprepitant as an adjunctive therapy to limit neuroinflammatory damage associated with bacterial central nervous system infections, particularly Lyme disease.
Its mechanism of action involves antagonizing the substance P (SP)/NK-1R interaction, which is associated with exacerbation of the central nervous system inflammatory response during infection.
In a non-human primate model, aprepitant reduced neuroinflammation induced by Borrelia burgdorferi, supporting further investigation into its potential for treating neuroinflammatory diseases. [1]

C23H21F7N4O3

534.426669836044

534.15

C, 51.69; H, 3.96; F, 24.88; N, 10.48; O, 8.98

170729-80-3

135413536

White solid powder

1.5±0.1 g/cm3

75-76 °C(lit.)

1.564

4.23

2

12

6

37

810

3

O=C1NC(CN2[C@H]([C@@H](O[C@@H](C3=CC(C(F)(F)F)=CC(C(F)(F)F)=C3)C)OCC2)C4=CC=C(F)C=C4)=NN1

ATALOFNDEOCMKK-OITMNORJSA-N

InChI=1S/C23H21F7N4O3/c1-12(14-8-15(22(25,26)27)10-16(9-14)23(28,29)30)37-20-19(13-2-4-17(24)5-3-13)34(6-7-36-20)11-18-31-21(35)33-32-18/h2-5,8-10,12,19-20H,6-7,11H2,1H3,(H2,31,32,33,35)/t12-,19+,20-/m1/s1

3-[[(2R,3S)-2-[(1R)-1-[3,5-bis(trifluoromethyl)phenyl]ethoxy]-3-(4-fluorophenyl)morpholin-4-yl]methyl]-1,4-dihydro-1,2,4-triazol-5-one

L754030; L-754030; L 754030; MK0869; MK 0869; MK-0869; ONO7436; ONO-7436; ONO 7436; Emend; Aponvie; Cinvanti; L-754030; Aprepitant; trade name: Emend

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO: ~107 mg/mL (~200.2 mM)
Ethanol: ~15 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.68 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 25.0 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: ≥ 2.5 mg/mL (4.68 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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