JAK1/2, STAT5[1]
Bax and Bcl-2 (mitochondrial pathway)[1]
Not specifically defined - acts as a bioactive compound with anticancer activity targeting the JAK/STAT5 signaling pathway.

SK-MEL-2 melanoma cells are inhibited in their ability to proliferate by picrocrocin. Following a 24-hour incubation period, the picrocrocin IC50 against SK-MEL-2 cells was 20 μM. Apoptosis and cell cycle arrest are the causes of picrocrocin's antiproliferative action. Furthermore, in SK-MEL-2 cells, picrocrocin can lower MMP levels and increase ROS levels. In conclusion, picrocrocin suppresses the SK-MEL-2 melanoma cells' JAK/STAT5 signaling pathway [1].

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Picrocrocin exhibited concentration-dependent growth inhibitory effects on SK-MEL-2 human malignant melanoma cells with an IC50 of 20 µM at 24 h incubation as determined by MTT assay (0-100 µM, 24 h)[1].
Picrocrocin induced apoptosis in SK-MEL-2 cells as shown by DAPI staining (0, 10, 20, 40 µM, 24 h), with apoptotic cell population increasing from 3.2% (control) to 56.5% at 40 µM dose, quantified by Annexin V/PI flow cytometry[1].
Picrocrocin triggered G2/M cell cycle arrest in SK-MEL-2 cells as determined by flow cytometric analysis after treatment with 0, 10, 20, 40 µM for 24 h, with the percentage of cells in G2 phase increasing dose-dependently[1].
Picrocrocin enhanced reactive oxygen species (ROS) levels in SK-MEL-2 cells up to 205% at 40 µM compared to control, measured by DCFH-DA staining and flow cytometry[1].
Picrocrocin decreased mitochondrial membrane potential (MMP) in SK-MEL-2 cells to 44% at 40 µM compared to control, measured by DiOC6 staining and flow cytometry[1].
Picrocrocin increased the protein expression of pro-apoptotic Bax and decreased anti-apoptotic Bcl-2 in a dose-dependent manner (0, 10, 20, 40 µM, 24 h) as assessed by western blotting[1].
Picrocrocin inhibited the phosphorylation of p-JAK1/2 and p-STAT5 in SK-MEL-2 cells without altering total JAK1/2 and STAT5 protein levels, as shown by western blotting[1].
In vitro studies demonstrate that Picrocrocin exhibits growth inhibitory effects against SKMEL-2 human malignant melanoma cells. Its mechanism of action involves targeting the JAK/STAT5 signaling pathway, which leads to cell cycle arrest and mitochondrial-mediated apoptosis. The compound's anticancer activity has been reported, though detailed mechanistic studies are limited. Picrocrocin is also studied for its role as a bioactive compound in saffron, contributing to its bitter taste and serving as a precursor to safranal.

In vivo studies on Picrocrocin are limited, as the compound is primarily studied for its role as a natural product and quality marker in saffron. The compound's anticancer activity has been demonstrated in vitro, suggesting potential for further in vivo evaluation in cancer models. However, detailed pharmacokinetic and efficacy data in animal models are not well characterized. The compound's natural occurrence in saffron and its role as a quality marker make it of interest for food and nutraceutical research, though therapeutic applications require further investigation.

For analytical chemistry applications, Picrocrocin is analyzed by high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC-MS) to determine its concentration in saffron extracts and to assess saffron quality. The compound is identified based on its retention time and mass spectrum. For biological activity studies, in vitro assays such as cell viability assays (MTT or CCK-8) are performed using cancer cell lines such as SKMEL-2 human malignant melanoma cells. Apoptosis is evaluated using Annexin V/PI staining, caspase activity assays, or Western blotting for apoptotic markers. JAK/STAT5 signaling is assessed by Western blotting for phosphorylated STAT5.

Cell viability assay (MTT): SK-MEL-2 cells were seeded at a density of 1×10^6 cells per well in 96-well plates and incubated for 12 h. The cells were then treated with varying concentrations of picrocrocin (0-100 µM) for 24 h. Subsequently, MTT solution (20 µL) was added to each well, followed by addition of 500 µL DMSO to solubilize formazan crystals. Absorbance was measured using an ELISA plate reader. The IC50 was determined as 20 µM[1].
Apoptosis assay (DAPI staining): SK-MEL-2 cells (1×10^6 per well in 6-well plates) were treated with 0, 10, 20, and 40 µM picrocrocin for 24 h. Cells were then stained with DAPI and examined under a fluorescence microscope to visualize apoptotic cells[1].
Apoptosis assay (Annexin V/PI flow cytometry): SK-MEL-2 cells were seeded in 6-well plates, treated with different doses of picrocrocin (0, 10, 20, 40 µM) for 24 h, harvested, washed with PBS, and then stained with Annexin V/FITC and propidium iodide (PI) for 20 min. The percentage of apoptotic cells was analyzed by flow cytometry[1].
Cell cycle analysis: Approximately 1×10^5 SK-MEL-2 cells per well in 6-well plates were allowed to adhere overnight, then treated with various doses of picrocrocin (0, 10, 20, 40 µM) for 24 h. Cell cycle phase distribution was determined by flow cytometry[1].
ROS assay: SK-MEL-2 cells (2×10^5 per well in 6-well plates) were treated with 0, 10, 20, 40 µM picrocrocin for 24 h at 37°C in 5% CO2. Cells were harvested, washed with PBS, resuspended in 500 µL of 10 µM DCFH-DA, and incubated for 30 min at 37°C in the dark. ROS levels were then measured by flow cytometry[1].
MMP assay: SK-MEL-2 cells prepared as above were resuspended in 500 µL of 1 µmol/L DiOC6 and incubated for 30 min at 37°C in the dark. Mitochondrial membrane potential (MMP) was assessed by flow cytometry[1].
Western blotting: SK-MEL-2 cells treated with picrocrocin were lysed, and protein concentration in each lysate was determined by BCA assay. Protein expression of Bax, Bcl-2, JAK1/2, p-JAK1/2, STAT5, and p-STAT5 was detected by western blotting following previously described protocols[1].
Cellular assays for Picrocrocin typically involve culturing cancer cell lines such as SKMEL-2 human malignant melanoma cells and treating them with the compound at various concentrations. Cell viability is assessed using MTT or CCK-8 assays. Apoptosis is evaluated using Annexin V/PI staining, caspase-3/7 activity assays, or Western blotting for apoptotic markers (cleaved PARP, cleaved caspase-3). Cell cycle analysis is performed by flow cytometry following propidium iodide staining. JAK/STAT5 signaling is assessed by Western blotting for phosphorylated STAT5 and total STAT5. Cytotoxicity against normal cells is assessed in parallel to evaluate selectivity.

In vivo efficacy of Picrocrocin is evaluated in animal models of cancer, though studies are limited. For anticancer studies, xenograft mouse models using melanoma or other cancer cell lines may be used to assess the compound's ability to inhibit tumor growth. Endpoints include tumor growth inhibition, assessment of apoptosis and proliferation markers in tumor tissues by immunohistochemistry, and evaluation of JAK/STAT5 signaling. Pharmacokinetic studies are needed to determine the compound's bioavailability, half-life, and tissue distribution. The compound's natural origin and presence in saffron suggest a potential favorable safety profile for nutraceutical applications.

Pharmacokinetic properties of Picrocrocin have been studied in the context of its role as a natural product and its presence in saffron. As a glycoside, Picrocrocin is likely metabolized in the gastrointestinal tract by gut microbiota or intestinal enzymes, releasing the aglycone. The compound's molecular weight of 330.37 and polar nature influence its absorption, distribution, metabolism, and excretion characteristics. Picrocrocin is soluble in water and alcohol, and slightly soluble in chloroform. Detailed pharmacokinetic data are limited, as the compound is not typically studied for pharmaceutical applications.

Toxicological evaluation of Picrocrocin is limited, as the compound is primarily studied as a natural product and food component. Saffron and its constituents, including Picrocrocin, have a long history of use in food and traditional medicine, suggesting a favorable safety profile at typical dietary levels. Standard toxicology assessments for pharmaceutical applications would include in vitro cytotoxicity assays, acute and repeated-dose toxicity studies, and genotoxicity assays. The compound's natural origin and presence in saffron support its potential safety for nutraceutical and research applications, though comprehensive toxicology data may be limited.

[1]. Picrocrocin exhibits growth inhibitory effects against SKMEL- 2 human malignant melanoma cells by targeting JAK/ STAT5 signaling pathway, cell cycle arrest and mitochondrial mediated apoptosis. J BUON. 2018 Jul-Aug;23(4):1163-1168.

Picrocrocin is a β-D-glucoside of β-cyclic citral and a precursor of saffron aldehyde. It is the main source of the bitterness in saffron. Its function is related to β-cyclic citral. Picrocrocin has been reported to be present in saffron (Crocus sativus), saffron (Crocus tommasinianus), and spring saffron (Crocus vernus), and relevant data are available for reference.

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Melanoma is one of the lethal types of skin malignancies responsible for significant morbidity and mortality worldwide. Current chemotherapy creates many adverse effects and results are often unsatisfactory. Picrocrocin as a saffron apocarotenoid has shown antiproliferative effects against various cancer cells (e.g., cervical cancer, breast cancer, gastric adenocarcinoma) in previous studies. In this study, picrocrocin induced apoptosis via mitochondrial pathway (altered Bax/Bcl-2 ratio, increased ROS, decreased MMP), triggered G2/M cell cycle arrest, and inhibited JAK/STAT5 signaling in SK-MEL-2 melanoma cells. The authors concluded that picrocrocin may be an important lead molecule for melanoma treatment, and further in vivo evaluation and semi-synthesis of more potent derivatives are required[1].
Picrocrocin is a naturally occurring monoterpene glycoside found in saffron that serves as a key quality marker and contributes to the spice's characteristic bitter taste and aroma. The compound is a degradation product of zeaxanthin and a precursor of safranal. Picrocrocin has been reported to exhibit anticancer activity against melanoma cells through targeting the JAK/STAT5 signaling pathway. The compound is not a drug and is not approved for therapeutic use, but it is studied as a bioactive natural product for its potential health benefits and as a quality marker for saffron authentication. Its primary applications are in food science, natural product research, and analytical chemistry.

C16H26O7

330.3734

330.168

C, 58.17; H, 7.93; O, 33.90

138-55-6

130796

White to off-white solid powder

1.31g/cm3

520.4ºC at 760mmHg

196 ºC (ethyl acetate )

187.1ºC

5.27E-13mmHg at 25°C

1.561

-0.5

4

7

4

23

473

6

O([C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[H])[C@]1([H])C([H])([H])C(C([H])([H])[H])=C(C([H])=O)C(C([H])([H])[H])(C([H])([H])[H])C1([H])[H]

WMHJCSAICLADIN-WYWSWGBSSA-N

InChI=1S/C16H26O7/c1-8-4-9(5-16(2,3)10(8)6-17)22-15-14(21)13(20)12(19)11(7-18)23-15/h6,9,11-15,18-21H,4-5,7H2,1-3H3/t9-,11-,12-,13+,14-,15-/m1/s1

(4R)-2,6,6-trimethyl-4-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycyclohexene-1-carbaldehyde

Picrocrocin; Picrocrocine; Saffron-bitter;

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 : ~100 mg/mL (~302.69 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.57 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 (7.57 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 (7.57 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.)