Human Endogenous Metabolite
- Protein Kinase C (PKC): Phytosphingosine inhibited approximately 90% of PKC activity at concentrations of 0.01%, 0.1%, and 0.2%. [1]
- G protein-coupled receptor 120 (GPR120): The IC50 value for Phytosphingosine-induced GPR120 activation was 33.4 µM. [4]
- Bax, Bcl-2, p53, PARP, Cytochrome C, Caspase-3, VEGF, CDK1: Molecular docking showed binding potential with these targets. Binding energies were: Bax (-5.46 kcal/mol), Bcl-2 (-4.72 kcal/mol), P53 (-6.48 kcal/mol), Parp (-3.39 kcal/mol), Cytochrome C (-4.50 kcal/mol), Caspase-3 (-7.21 kcal/mol), VEGF (-4.83 kcal/mol), and CDK1 (-6.31 kcal/mol). [2]

- Anti-microbial activity: Phytosphingosine inhibited the growth of Gram-positive bacteria (Micrococcus luteus, Staphylococcus aureus, Corynebacterium xerosis, Propionibacterium acnes), Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli), Yeasts (Candida albicans), and Dermatophytes (Microsporum canis). The concentration required for growth inhibition of P. acnes within 1 hour was 0.020%. [1]
- Inhibition of IL-1α release: In UVB-irradiated human skin explants, Phytosphingosine at 0.2% and 1% inhibited the release of IL-1α by 78% and 72%, respectively (P < 0.05). [1]
- Inhibition of PKC activity: Phytosphingosine at concentrations of 0.01%, 0.1%, and 0.2% inhibited approximately 90% of PKC activity. No inhibitory effect was detected at 0.001%. [1]
- Effect on irritated artificial human epidermis: In a 3-D artificial skin model irritated with SDS, a formulation containing 0.15% Phytosphingosine reduced LDH release (from ~19 UL⁻¹ to ~10 UL⁻¹) and reduced IL-1α release (from ~35 pg/mL to 22 pg/mL). [1]
- Cytotoxicity on lung cancer cells: Phytosphingosine significantly inhibited the proliferation of A549 and LLC cells in a concentration- and time-dependent manner. The IC50 values for A549 and LLC cells at 24h were 4.3 ± 0.5 μg/mL and 4.5 ± 1.2 μg/mL, respectively. The IC50 for normal BEAS-2B cells was 6.3 ± 1.1 μg/mL. [2]
- Clonogenic assay: Phytosphingosine treatment (1, 3, 5 μg/mL for 24h) significantly reduced the clone number in A549 cells (p < 0.001), producing a long-term anti-proliferative activity. [2]
- Induction of apoptosis: Phytosphingosine dose-dependently induced apoptosis in A549 cells. It caused chromatin condensation and fragmentation. [2]
- Effect on apoptosis-related proteins: Phytosphingosine treatment of A549 cells upregulated Bax and downregulated Bcl-2, increasing the Bax/Bcl-2 ratio. It also promoted cytochrome c release and upregulated caspase 9 and caspase 3, leading to PARP cleavage. [2]
- Cell cycle arrest: Phytosphingosine (1-5 μg/mL for 24h) induced G2/M phase arrest in A549 cells. The proportion of G2/M phase cells increased from 12.48% to 18.19%. It downregulated Cyclin B1 and CDK1 protein levels. [2]
- ROS production: Phytosphingosine significantly induced an increase in reactive oxygen species (ROS) levels in A549 cells. This effect was reduced by pre-treatment with N-Acetyl-L-cysteine (NAC). [2]
- Mitochondrial membrane potential (MMP): Phytosphingosine decreased the mitochondrial membrane potential of A549 cells. The percentage of red fluorescent positive cells decreased from 86.35% to 1.2%, and green fluorescent positive cells increased from 11.22% to 87.6%. [2]
- Anti-fungal activity: Phytosphingosine (80 µM) inhibited mycelium growth of plant-interacting fungi Sclerotinia sclerotiorum, Fusarium graminearum, Verticillium longisporum, and Serendipita indica on agar media. Mycelial area was reduced by 90% (S. indica), 85% (S. sclerotiorum), 49% (V. longisporum), and 16% (F. graminearum). It also reduced fungal biomass production in liquid media. [3]
- Anti-bacterial activity: Phytosphingosine at 15.9 μg/mL killed 95% of Pseudomonas syringae pv. tomato, Agrobacterium tumefaciens, and Rhizobium radiobacter F4 within 15-20 min. 50 µM of Phytosphingosine substantially reduced Pst growth over time, while 100 µM completely abolished growth for 48h. The phosphorylated form (t18:0-P) was not effective. [3]
- GPR120 activation: In a TGFα-shedding assay using 293T cells expressing GPR120, Phytosphingosine activated GPR120. The IC50 value was 33.4 μM. Phytosphingosine also induced an increase in intracellular Ca²⁺ levels and increased Erk phosphorylation. Activation was inhibited by the specific antagonist AH7614. Dihydrosphingosine (DHS) also strongly activated GPR120, while sphingosine did not. [4]

- Anti-acne clinical study (Group 1): In a 60-day, half-face trial on patients with moderate inflamed acne, comparing benzoyl peroxide (BPO) vs. BPO + 0.2% Phytosphingosine-HCl (PS-HCl). At day 60, BPO reduced comedones by 22%, while the combination reduced them by 72%. BPO reduced papules/pustules by 32%, while the combination reduced them by 88%. [1]
- Anti-acne clinical study (Group 2): In a 60-day, half-face trial comparing placebo vs. 0.2% Phytosphingosine-HCl. Placebo increased comedones by 43%, while PS-HCl limited the increase to 6%. PS-HCl reduced papules/pustules by 89% at day 60, while placebo had no effect. [1]
- Anti-lung cancer activity in mice: In a C57BL/6 mouse model bearing LLC tumors, Phytosphingosine (0.5, 1, 2 mg/kg, given every other day for 10 times) significantly suppressed tumor cell growth. A significant decrease in body weight was observed in the 2 mg/kg group. The cisplatin group (5 mg/kg, every 5 days for 4 times) also showed toxicity. [2]
- Histopathology: H&E staining showed areas of necrosis in tumor tissue from Phytosphingosine-treated mice. In the high-dose group (2 mg/kg), pathological changes with congestion were observed in kidney tissue. [2]
- Effect on immune cells in mice: Phytosphingosine treatment (2 mg/kg) significantly decreased the number of T cells (CD3+ cells) in the spleen of LLC tumor-bearing mice. The number of myeloid-derived suppressor cells (MDSCs) in the spleen was significantly reduced in the Phytosphingosine-treated group (p < 0.001). [2]
- Anti-bacterial activity in plants: Co-infiltration of Phytosphingosine (100 µM) with Pseudomonas syringae pv. tomato (Pst) into Arabidopsis leaves reduced disease symptoms and host tissue damage. Co-infiltration prevented an increase in bacterial DNA per plant DNA 24 and 48 hours post-infiltration, indicating growth inhibition of Pst in the leaf. [3]
- GPR120 activation in vivo: Oral administration of Phytosphingosine was previously found to improve glucose tolerance in high-fat diet-induced mice. [4]

- PKC Activity Assay: Pure PKC was incubated with a substrate (histone) and radiolabelled adenosine-3-phosphate (ATP) in the presence and absence of varying concentrations of Phytosphingosine (range 0.001 - 0.2%). The reference product staurosporine (10 µM) was used as a control. The incorporated radioactivity in the radiolabelled reaction product was measured by liquid scintillation. The degree of inhibition by staurosporine and Phytosphingosine-HCl compared to the control was calculated based on the amount of phosphorylated histone produced. [1]
- Molecular Docking: The chemical structures of Phytosphingosine were drawn and optimized. Protein sequences and structures were obtained from the Protein Data Bank. The Autodock tool was used to process receptor protein files. The mol2 file with information about Phytosphingosine was used for molecular docking. The docking results were visualized using pymol to explore interaction with potential targets like Bax, Bcl-2, p53, PARP, cytochrome C, caspase-3, VEGF, and CDK1. [2]

- IL-1α Release Assay: Human skin explants in culture were used. Phytosphingosine (0.2% and 1.0%) and dexamethasone (10⁻⁶ M) were applied to the skin explants 1 hour before and immediately after UVB irradiation (20 min of UVB 2 J/cm²). Products were rinsed off before irradiation. The interleukin-1α secretion was measured after 24 hours using an ELISA kit. The baseline was defined as IL-1α in non-treated, non-exposed skin; maximal IL-1α production was from non-treated, UVB-exposed skin. [1]
- Cell Viability (MTT) Assay: Cells (A549, LLC, BEAS-2B) were seeded into 96-well plates and incubated for 24h before treatment with different concentrations of Phytosphingosine. Cisplatin (30 μg/mL) was used as a positive control. After 24h and 48h, MTT solution was added and incubated for 4h. The formazan products were dissolved in DMSO, and the absorbance was measured at 490 nm. Data was presented as a percentage of the control group. [2]
- Clonogenic Assay: A549 cells were treated with Phytosphingosine (1, 3, 5 μg/mL) for 24h, then the supernatant was discarded and cells were washed. Cells were collected, centrifuged, resuscitated, and inoculated in a 6-well plate. After culturing continuously for 21 days, cells were fixed with paraformaldehyde and stained with crystal violet. Clone numbers containing more than 50 cells were counted under a light microscope. [2]
- Apoptosis Analysis: A549 cells were treated with Phytosphingosine for 24h and stained using an Annexin V-FITC/propidium iodide (PI) apoptosis detection kit. Cells were resuspended with binding buffer, and Annexin V-FITC and PI were added. After incubation in the dark, stained cells were analyzed by flow cytometry. [2]
- Cell Cycle Analysis: A549 cells were harvested after Phytosphingosine treatment for 24h, fixed in cold 70% ethanol, and then stained with PI. Samples were analyzed by flow cytometry to determine the distribution of the cell cycle. [2]
- Hoechst 33258 Staining: A549 cells were treated with Phytosphingosine (1, 3, 5 μg/mL) for 24h, then fixed with ice-cold paraformaldehyde. After washing with PBS, cells were stained with Hoechst 33258 and observed by fluorescence phase-contrast microscope to examine nuclear morphological changes. [2]
- Mitochondrial Membrane Potential (MMP) Assay: A549 cells were treated with Phytosphingosine for 24h, washed with PBS, and stained with the JC-1 fluorescent probe. Samples were analyzed by flow cytometry and using an inverted fluorescence microscope. [2]
- Intracellular ROS Detection: A549 cells were treated with Phytosphingosine for 24h, washed with PBS, and stained with the fluorescent probe DCFH-DA. Samples were analyzed by flow cytometry. In some experiments, cells were pre-treated with N-acetyl-L-cysteine (NAC) for 1h before Phytosphingosine treatment. [2]
- Western Blot: Proteins were extracted from A549 cells treated with different concentrations of Phytosphingosine for 24h. Primary antibodies against β-actin, Bax, Bcl-2, cytochrome C, cyclin B1, CDK1, caspase 3, cleaved PARP, and caspase 9 were used. HRP-conjugated secondary antibodies and ECL substrate were used for detection. [2]
- GPR120 TGFα-Shedding Assay: 293T cells were seeded in a 6-well plate and transfected with plasmids encoding hGPR120 and AP-TGFα. Cells were then re-seeded in a 96-well plate. Phytosphingosine or other test substances were added and cultured. The AP activity in the supernatant and cells was determined using p-nitro-phenyl phosphate (pNPP) as a substrate. The data represents the ratio of AP activity in the supernatant to total AP activity, expressed as a percentage. [4]
- Intracellular Ca²⁺ Measurement: 293T cells were cultured with serum-free medium and then incubated with the Calcium Kit-Fluo 4 loading buffer. The buffer was changed to a recording medium. Phytosphingosine or ALA was added, and the time change of fluorescence intensity (Ex 488 nm, Em 510 nm) was measured. [4]

- Topical in vivo anti-microbial study: The anti-microbial efficacy of topical Phytosphingosine within an emulsion-based formulation was tested on the unwashed hands of 12 subjects. Formulations containing Phytosphingosine or its salt PS-HCl were compared to a control formulation and triclosan (positive control). The total microbial count was determined on the skin at t = 0, after 1h, and after 4h. Percentages from baseline were calculated. [1]
- Clinical study on acne skin (Randomized, half-face trial):
Part 1: 15 men and 15 women (average age 20) with moderate inflamed acne applied benzoyl peroxide (4%) on one side of the face and a combination of 0.2% Phytosphingosine-HCl with benzoyl peroxide on the other side, twice daily for 60 days.
Part 2: 10 volunteers applied a placebo preparation on one side of the face and 0.2% Phytosphingosine-HCl on the other side, twice daily for 60 days. Patients used a purifying gel as a cleansing base. Dermatologists evaluated comedones, papules, and pustules on day 0, 30, and 60. [1]
- In vivo anti-lung cancer study in mice: Female C57BL/6 mice were inoculated with LLC cell suspension (1 × 10⁶ cells/mouse) to establish a lung cancer model. After 7 days, mice with 15-30 mm³ tumors were randomly divided into 6 groups (6 mice per group): control, DMSO, cisplatin, and Phytosphingosine (0.5, 1, 2 mg/kg). Phytosphingosine was given every other day for 10 times. Cisplatin (5 mg/kg) was injected intraperitoneally every 5 days for 4 times. Tumor volumes were measured using calipers and calculated as (length × width²)/2. On day 28, mice were executed, and organs were dissected for index calculation, histology (H&E staining), and biochemical assays. [2]
- Plant co-infiltration experiment: Pseudomonas syringae pv. tomato (Pst) strains were diluted to OD 0.01 in 10 mM MgCl₂ containing 100 µM Phytosphingosine, 100 µM phytosphingosine-1-phosphate, or solvent only. Leaves of Arabidopsis thaliana were infiltrated with the bacterial solutions from the abaxial side. Photographs were taken 72h after infiltration to assess leaf damage. Cell death was quantified using leaf disc ion leakage assays, where leaf discs were floated on ultrapure H₂O, and conductivity was measured over 24h. [3]

- Phytosphingosine is present in small amounts in humans and is found in the small intestine and skin. In humans, it is synthesized by a desaturase called DES2.
Using DES2 knockout mice, it was found that dietary Phytosphingosine is incorporated and reaches the liver, indicating that in addition to being synthesized by the human body, Phytosphingosine of dietary origin can also be used. [4]

- Cutaneous tolerance: In the clinical acne study, no case of cutaneous intolerance was indicated by any subject for the combination of benzoyl peroxide with Phytosphingosine-HCl or the pure PS-HCl preparation. [1]
- Organ toxicity in mice: In the lung cancer mouse model, the cisplatin group showed a significant decrease in spleen index, an increase in kidney and liver index, indicating toxicity. The high dose of Phytosphingosine (2 mg/kg) showed a significant decrease in spleen index and an increase in kidney organ index (p < 0.05). H&E staining revealed pathological changes with congestion in the kidney tissue of the high-dose group. [2]
- Blood biochemistry in mice: Blood biochemical tests showed that the cisplatin group caused damage to the liver (increased ALT, p < 0.01) and kidneys (increased BUN and Cr, p < 0.01). In the high-dose Phytosphingosine group (2 mg/kg), there was an increase in creatinine (Cr) (p < 0.05), indicating kidney damage. There was no significant change in ALT and AST indices in the high-dose group, suggesting no effect on the liver. [2]

[1]. Anti‐microbial and‐inflammatory activity and efficacy of phytosphingosine: an in vitro and in vivo study addressing acne vulgaris. International journal of cosmetic science, 2007, 29(3): 181-190.

[2]. Phytosphingosine inhibits the growth of lung adenocarcinoma cells by inducing G2/M-phase arrest, apoptosis, and mitochondria-dependent pathway cell death in vitro and in vivo. Chem Biol Interact. 2024 Jan 5;387:110795.

[3]. The major plant sphingolipid long chain base phytosphingosine inhibits growth of bacterial and fungal plant pathogens. Sci Rep. 2022 Jan 20;12(1):1081.

[4]. Phytosphingosine is a novel activator of GPR120. J Biochem. 2018 Jul 1;164(1):27-32.

Phytosphingosine is a sphingosine compound belonging to the amino alcohol and triol class. It is a metabolite of Saccharomyces cerevisiae and mice. Its function is related to N-acyl-β-D-galactosyl phytosphingosine and sphingosine. It is the conjugate base of phytosphingosine (1+). Phytosphingosine has been reported in Klebsiella pneumoniae, cattle, and other organisms with relevant data. Phytosphingosine is a metabolite found or produced in Saccharomyces cerevisiae. 4-Hydroxysphingosine is a metabolite found or produced in Saccharomyces cerevisiae. See also: Phytosphingosine hydrochloride (active moiety); Salicyloyl phytosphingosine (active moiety); Nicotinamide; Phytosphingosine; Salicylic acid (component).

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- Role in acne: The anti-microbial and anti-inflammatory activities of Phytosphingosine work synergistically, resulting in a good effect on the clinical status of acne-prone individuals, particularly for inflammatory manifestations. It can enhance or complement existing acne therapies. [1]
- Role in lung cancer: Phytosphingosine induces apoptosis through a mitochondria-mediated pathway, arrests the cell cycle at the G2/M phase, and damages mitochondrial functions, increasing ROS levels. It may be a potential candidate for the treatment of lung cancer. [2]
- Role in plant defence: Elevated levels of Phytosphingosine in plants may be a mechanism to protect against pathogens like Pseudomonas syringae. Production and export of t18:0 into the apoplast could be a mechanism to restrict pathogen growth in the leaf. Phytosphingosine was detected in wheat root exudates, suggesting plants may employ it to influence microbial growth. [3]
- Role in diabetes: Phytosphingosine is a novel ligand for GPR120, superior to the synthetic ligand GW9508. GPR120 activation promotes incretin secretion and increases brown adipocytes/beige cells, which can suppress obesity and improve type II diabetes. Phytosphingosine is abundant in yeast-containing foods like fermented foods, and dietary intake may have anti-diabetic effects. [4]

C18H39NO3

317.50716

317.292

554-62-1

122121

White to off-white solid

1.0±0.1 g/cm3

483.7±40.0 °C at 760 mmHg

102ºC

246.4±27.3 °C

0.0±2.8 mmHg at 25°C

1.490

5.18

4

4

16

22

226

3

CCCCCCCCCCCCCC[C@H]([C@H]([C@H](CO)N)O)O

AERBNCYCJBRYDG-KSZLIROESA-N

InChI=1S/C18H39NO3/c1-2-3-4-5-6-7-8-9-10-11-12-13-14-17(21)18(22)16(19)15-20/h16-18,20-22H,2-15,19H2,1H3/t16-,17+,18-/m0/s1

(2S,3S,4R)-2-aminooctadecane-1,3,4-triol

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 : ~50 mg/mL (~157.48 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.87 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 (7.87 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 25.0 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: ≥ 2.5 mg/mL (7.87 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.)