PKC:660 nM (IC50); PKA:0.17 mM (IC50); TPK:0.1 mM (IC50)
Chelerythrine displaces the BH3-containing protein Bax from BclXL and inhibits the BclXL-Bak BH3 peptide binding with an IC50 of 1.5 μM. Loaded cells treated with chelerythrine underwent cell engraftment, which was characterized by mitochondrial involvement [1]. Chelidonine treatment activates and inhibits LPS-induced TNF-α levels and LPS-induced mouse peritoneal giant cells by blocking p38 mitogen protein activator (MAPK) and extracellular signal regulatory protein activator 1 and 2 (ERK1/2). Furthermore, the control of intermediate mediator expression by p38 MAPK and ERK1/2 may account for the effects of chelerythrine on the generation of NO and the cytokine TNF-α [2]. With an LD50 value of 3.46 μM, erythrine is cytotoxic to human monocyte leukocytes. Cells exposed to sanguinarine and chelerythrine dramatically decreased the expression of CCL-2 by 3.5 and 1.9 times, respectively, two hours after LPS stimulation [3]. In a dose-dependent way, chelidonine chloride markedly increased ERK1/2 phosphorylation. Furthermore, p38 phosphorylation can be inhibited by chelerythrine chloride [4].

Chelerythrine displaces the BH3-containing protein Bax from BclXL and inhibits the BclXL-Bak BH3 peptide binding with an IC50 of 1.5 μM. Loaded cells treated with chelerythrine underwent cell engraftment, which was characterized by mitochondrial involvement [1]. Chelidonine treatment activates and inhibits LPS-induced TNF-α levels and LPS-induced mouse peritoneal giant cells by blocking p38 mitogen protein activator (MAPK) and extracellular signal regulatory protein activator 1 and 2 (ERK1/2). Furthermore, the control of intermediate mediator expression by p38 MAPK and ERK1/2 may account for the effects of chelerythrine on the generation of NO and the cytokine TNF-α [2]. With an LD50 value of 3.46 μM, erythrine is cytotoxic to human monocyte leukocytes. Cells exposed to sanguinarine and chelerythrine dramatically decreased the expression of CCL-2 by 3.5 and 1.9 times, respectively, two hours after LPS stimulation [3]. In a dose-dependent way, chelidonine chloride markedly increased ERK1/2 phosphorylation. Furthermore, p38 phosphorylation can be inhibited by chelerythrine chloride [4].

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Chelerythrine (CHE) significantly inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO) production in murine peritoneal macrophages in a dose-dependent manner (at concentrations of 10⁻⁵, 10⁻⁴, 10⁻³, 10⁻², and 10⁻¹ µg/ml). The inhibition was comparable to that achieved by specific p38 MAPK inhibitor SB203580 (10 µM) and ERK1/2 inhibitor PD98059 (20 µM). [1]
Chelerythrine (CHE) significantly decreased lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-α) production in murine peritoneal macrophages in a dose-dependent manner (at concentrations of 10⁻⁵, 10⁻⁴, 10⁻³, 10⁻², and 10⁻¹ µg/ml) over time (4, 6, 12, and 24 h). The inhibition was comparable to that achieved by specific p38 MAPK inhibitor SB203580 (10 µM) and ERK1/2 inhibitor PD98059 (20 µM). [1]
Chelerythrine (CHE) significantly suppressed lipopolysaccharide (LPS)-induced phosphorylation of ERK1/2 and p38 MAPK in murine peritoneal macrophages in a dose-dependent manner (at concentrations of 10⁻⁴, 10⁻², and 10⁻¹ µg/ml), as determined by Western blot analysis. The inhibition was comparable to that achieved by specific inhibitors SB203580 and PD98059. [1]
Chelerythrine (CHE) exhibited cytotoxic effects on murine peritoneal macrophages at concentrations of 1 and 10 µg/ml after 24-hour treatment, as assessed by MTT assay. Concentrations up to 10⁻¹ µg/ml did not significantly affect cell viability. [1]

Chelerythrine inhibits the production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α) in the serum when exposed to LPS, thereby exhibiting significant anti-inflammatory effects in an in vivo experimentally induced mouse endotoxic shock model [2]. When administered intraperitoneally, chelerythrine chloride (5 mg/kg/day) causes RCC cells to undergo apoptosis without causing considerable harm to mice. P53 accumulates in response to cholerylthrine chloride treatment in a dose-dependent manner [4].

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Chelerythrine (CHE) pretreatment (intraperitoneal injection at doses of 1, 5, or 10 mg/kg, administered 24 and 1 hour before LPS) significantly increased the survival rate of mice in an LPS-induced endotoxic shock model in a dose-dependent manner. The 10 mg/kg dose provided full protection (8/8 mice survived), 5 mg/kg offered 75% protection (6/8 survived), and 1 mg/kg offered 37.5% protection (3/8 survived) within 72 hours. [1]
Chelerythrine (CHE) pretreatment (intraperitoneal injection at doses of 1, 5, or 10 mg/kg) significantly suppressed the LPS-induced increase in serum nitric oxide (NO) levels at 3 and 6 hours post-LPS injection in a dose-dependent manner. [1]
Chelerythrine (CHE) pretreatment (intraperitoneal injection at doses of 1, 5, or 10 mg/kg) significantly decreased the LPS-induced elevation of serum tumor necrosis factor-alpha (TNF-α) levels at 1, 3, and 6 hours post-LPS injection in a dose-dependent manner. [1]

The identification of small molecule inhibitors of antiapoptotic Bcl-2 family members has opened up new therapeutic opportunities, while the vast diversity of chemical structures and biological activities of natural products are yet to be systematically exploited. Here we report the identification of chelerythrine as an inhibitor of BclXL-Bak Bcl-2 homology 3 (BH3) domain binding through a high throughput screening of 107,423 extracts derived from natural products. Chelerythrine inhibited the BclXL-Bak BH3 peptide binding with IC50 of 1.5 micro m and displaced Bax, a BH3-containing protein, from BclXL. Mammalian cells treated with chelerythrine underwent apoptosis with characteristic features that suggest involvement of the mitochondrial pathway. While staurosporine, H7, etoposide, and chelerythrine released cytochrome c from mitochondria in intact cells, only chelerythrine released cytochrome c from isolated mitochondria. Furthermore BclXL-overexpressing cells that were completely resistant to apoptotic stimuli used in this study remained sensitive to chelerythrine. Although chelerythrine is widely known as a protein kinase C inhibitor, the mechanism by which it mediates apoptosis remain controversial. Our data suggest that chelerythrine triggers apoptosis through a mechanism that involves direct targeting of Bcl-2 family proteins[5].

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In Vitro Binding Analysis—Labeled Bax was prepared by in vitro transcription/translation of pXJHA-Bax using the TnT T7-coupled reticulocyte lysate system from Promega. The GST binding assay was performed as described previously except that increasing concentrations of chelerythrine were incubated with GSTBclXLΔ19 30 min prior to the addition [35S]Bax.[5]

Cell viability is evaluated via MTT assay. Cells (2×103 HEK-293 cells/well and 3×103 SW-839 cells/well) in 100 µL medium are seeded into 96-well plates, and incubated for 12 h. Next, the medium in each well is replaced with medium containing various concentrations of Chelerythrine Chloride, and the cells are incubated at 37°C for an additional 24 and 48 h. Subsequently, 20 µL MTT (5 mg/mL) is added to each well. Following an additional incubation at 37°C for 4 h, the supernatant is removed, and 100 µL DMSO is added to each well. The absorbance values (read at 540 nm) are determined using the iMark™ Microplate Absorbance Reader[1].

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Previous studies have demonstrated that the benzo[c]phenanthridine alkaloid chelerythrine chloride (CC) has inhibitory effects on various tumors. However, the anticancer activity of CC and its underlying mechanisms have not been elucidated in renal cancer cells. The present study examined the effects of CC on growth inhibition and apoptosis of renal cancer cells in vitro and in vivo. Flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays revealed that CC markedly suppressed the growth of HEK-293 and human renal cancer SW-839 cells in a time- and dose-dependent manner. The xenograft mouse model, which was performed in nude mice, exhibited a reduced tumor growth following CC treatment. In addition, the present study revealed that CC significantly decreased the phosphorylation of extracellular signal-regulated kinase (ERK) and Akt, which was accompanied by upregulation of p53, B-cell lymphoma 2 (Bcl-2)-associated X protein, cleaved caspase-3 and cleaved poly (adenosine diphosphate-ribose) polymerase (PARP), and downregulation of Bcl-2, caspase-3 and PARP. Furthermore, the use of PD98059, a specific mitogen-activated protein kinase kinase inhibitor, potentiated the proapoptotic effects of CC, which indicated that CC may induce apoptosis in renal cancer cells partly via inhibition of ERK activity. Overall, the results of the present study demonstrated that CC may be developed as a potential anticancer treatment for patients with renal cancer[3].

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Cell Viability Assay (MTT Assay): Peritoneal macrophages were isolated from mice injected with thioglycolate. Cells were seeded in 96-well plates and treated with various concentrations of Chelerythrine (CHE) (10⁻⁵ to 10 µg/ml) alone or in combination with LPS (10 µg/ml) for 24 hours. After incubation, MTT reagent was added to each well. Following a 4-hour incubation, the formed formazan crystals were dissolved using DMSO. The absorbance was measured at 490 nm to assess cell viability. [1]
Nitric Oxide (NO) Measurement: Peritoneal macrophages were seeded in 24-well plates and pretreated with various concentrations of Chelerythrine (CHE) (10⁻⁵ to 10⁻¹ µg/ml), SB203580 (10 µM), or PD98059 (20 µM) for 24 hours, followed by exposure to LPS (10 µg/ml) for 12 hours. The culture supernatant was collected and mixed with Griess reagent. After incubation at room temperature in the dark, the absorbance was measured at 540 nm. Nitrite concentration was determined using a sodium nitrite standard curve. [1]
Tumor Necrosis Factor-alpha (TNF-α) ELISA: Peritoneal macrophages were pretreated with various concentrations of Chelerythrine (CHE) (10⁻⁵ to 10⁻¹ µg/ml) for 24 hours, followed by exposure to LPS (10 µg/ml) for 4, 6, 12, or 24 hours. The culture supernatant was collected. TNF-α levels were measured using a commercial ELISA kit according to the manufacturer's instructions. [1]
Western Blot Analysis: Peritoneal macrophages were pretreated with various concentrations of Chelerythrine (CHE) (10⁻⁴, 10⁻², 10⁻¹ µg/ml), SB203580 (10 µM), or PD98059 (20 µM) for 24 hours, followed by exposure to LPS (10 µg/ml) for 30 minutes. Total cell protein was extracted using lysis buffer. Protein concentration was determined. Equal amounts of protein were separated by SDS-PAGE and transferred to a membrane. The membrane was blocked and then incubated with primary antibodies against phospho-p38, phospho-ERK1/2, and GAPDH overnight. After washing, the membrane was incubated with a horseradish peroxidase-conjugated secondary antibody. Protein bands were visualized using an enhanced chemiluminescence system. [1]

Dissolved in PBS; 5 mg/kg; i.p.Mice bearing SQ-20B xenografts A total of 5×106 SW-839 cells are mixed with Matrigel®, and injected subcutaneously into the flanks of 14 5-week-old male BALB/c nude mice. The mice are maintained in 18×30-cm cages containing three mice each, at a temperature of 22°C using a 12 h light/dark cycle. Food and water is available ad libitum. The mice are randomLy divided into two groups (n=7). As previously described, the mice are administrated with chelerythrine chloride at a dose of 5 mg/kg/day via intraperitoneal injection for 5 weeks, with the first injection of chelerythrine chlorideurring 24 h after injection with the SW-839 cells. The control mice are administered with the same volume of PBS containing 1% DMSO. The volume and weight of the mouse tumors are measured once a week. All the mice are sacrificed 36 days subsequent to inoculation of the cancer cells, when the tumors are resected.[1]

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Acute Toxicity Study: Healthy male mice were administered Chelerythrine (CHE) intraperitoneally at an initial dose of 1 mg/kg. Mice were observed for toxic symptoms, general behavior, and mortality for up to 72 hours. Body weights were measured daily. If no toxicity or mortality was observed, the procedure was repeated with higher doses (5, 10, and 15 mg/kg). In this study, a dose of 10 mg/kg did not produce any toxic signs. [1]
LPS-Induced Endotoxic Shock Model: Male mice were divided into groups. The treatment groups received two intraperitoneal injections of Chelerythrine (CHE) at various doses (1, 5, or 10 mg/kg) at 24 and 1 hour before an intraperitoneal injection of LPS (100 µg/kg). The control group received vehicle, and the LPS group received LPS only. Blood samples were collected at 0, 1, 3, and 6 hours after LPS injection via the venous sinus. Serum was separated by centrifugation and stored at -80°C for analysis of TNF-α and NO levels. Survival was monitored for 72 hours. [1]
Peritoneal Macrophage Isolation for Cell Assays: Female mice were injected intraperitoneally with a 1% sodium thioglycolate solution. Six days later, mice were euthanized, and peritoneal macrophages were harvested by lavaging the peritoneal cavity with phosphate-buffered saline (PBS). Cells were collected by centrifugation, washed, and cultured in RPMI-1640 medium supplemented with fetal bovine serum and antibiotics. [1]

Toxicity Overview
Chelitaxel is a potent, selective, and cell-membrane-penetrating protein kinase C (PKC) inhibitor. It is also the main active natural product of the plant Zanthoxylum clava-herculis, exhibiting antibacterial activity against Staphylococcus aureus. (Wikipedia) Chelitaxel is a selective inhibitor of group A and group B PKC subtypes and possesses antitumor activity. Chelitaxel chloride inhibits PKC by activating neutral sphingomyelinase, ceramide accumulation, and sphingomyelin depletion, inducing apoptosis. Chelitaxel's selectivity for PKC is at least 100 times higher than other kinases. Chelitaxel competes with conserved catalytic sites of PKC and appears to be a potent and specific inhibitor of group A and group B kinases. In vitro experiments have shown that chelitaxel has cytotoxic activity against nine human tumor cell lines. In in vitro experiments, radioresistant and chemoresistant head and neck squamous cell carcinoma (HNSCC) cell lines rapidly underwent apoptosis after treatment with chelitaxel. Treatment of nude mice bearing SQ-20B HNSCC cells with Chelerythrine significantly delayed tumor growth. Furthermore, the toxicity of Chelerythrine treatment was extremely low. The subcutaneous LD50 in mice was 95 mg/kg, Planta Medica., 43(161), 1981 [PMID:7312984]. The intravenous LD50 in mice was 18500 μg/kg, Planta Medica., 43(161), 1981 [PMID:7312984]. In an acute toxicity study in mice, intraperitoneal injection of 10 mg/kg of Chelerythrine (CHE) resulted in no changes in general appearance or toxic symptoms observed during a 72-hour observation period. [1]

[1]. Effect of Chelerythrine Against Endotoxic Shock in Mice and Its Modulation of Inflammatory Mediators in Peritoneal Macrophages Through the Modulation of Mitogen-Activated Protein Kinase (MAPK) Pathway. Inflammation. 2012 Dec;35(6):1814-24. d

[2]. Investigation of sanguinarine and chelerythrine effects on LPS-induced inflammatory gene expression in THP-1 cell line. Phytomedicine. 2012 Jul 15;19(10):890-5. Epub 2012 May 14.

[3]. Chelerythrine chloride induces apoptosis in renal cancer HEK-293 and SW-839 cell lines. Oncol Lett. 2016 Jun;11(6):3917-3924.

[4]. Chelerythrine is a potent and specific inhibitor of protein kinase C. Biochem Biophys Res Commun. 1990 Nov 15;172(3):993-9.

[5]. Identification of chelerythrine as an inhibitor of BclXL function.J Biol Chem. 2003 Jun 6;278(23):20453-6.

[6]. Induction of reactive oxygen species-stimulated distinctive autophagy by chelerythrine in non-small cell lung cancer cells.Redox Biol. 2017 Aug;12:367-376.

Chelerythrine chloride is a benzo[c] alkaloid. Chelerythrine (CHE), a quaternary ammonium benzo[c] alkaloid, is a traditional Chinese medicine used to treat various inflammatory diseases. To gain a deeper understanding of the anti-inflammatory effects and molecular mechanisms of CHE, we investigated its anti-inflammatory function using an experimentally induced mouse endotoxin shock model and a lipopolysaccharide (LPS)-induced mouse peritoneal macrophage model. The results showed that CHE exhibited significant anti-inflammatory effects in the in vivo experimentally induced mouse endotoxin shock model, through the inhibition of LPS-induced serum tumor necrosis factor-α (TNF-α) levels and nitric oxide (NO) production. Furthermore, our data indicated that CHE treatment inhibited LPS-induced TNF-α levels and NO production in LPS-induced mouse peritoneal macrophages by selectively inhibiting the activation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases 1 and 2 (ERK1/2). In addition, the effects of CHE on NO and TNF-α production may be related to the role of p38 MAPK and ERK1/2 in regulating the expression of inflammatory mediators. [1]

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Quaternary ammonium benzo[c]phenanthridine alkaloids sanguisorbin and Chelerythrine are used in folk medicine due to their wide range of medicinal properties. One of their main functions is anti-inflammatory activity, but their specific mechanisms have not been fully elucidated. This study focuses on the ability of these alkaloids to regulate the gene expression of pro-inflammatory cytokines tumor necrosis factor α (TNF-α), monocyte chemoattractant protein 1 (MCP-1, also known as CCL-2), interleukin (IL)-6, IL-1β, and anti-inflammatory cytokines IL-1 receptor antagonist (IL-1RA) and IL-10. The effects of these alkaloids were compared with those of the traditional drug prednisone. Human monocyte-derived macrophages were pretreated with alkaloids or prednisone and then induced with lipopolysaccharide. The gene expression changes at the transcriptional levels of the above cytokines were detected. This study found that the main pro-inflammatory cytokines affected were CCL-2 and IL-6. Two hours after LPS stimulation, CCL-2 expression was significantly reduced in cells treated with sanguisorbin and Chelerythrine, decreasing by 3.5-fold (p<0.001) and 1.9-fold (p<0.01), respectively; while in prednisone-treated cells, CCL-2 expression decreased by 5.3-fold (p<0.001). Eight hours after LPS induction, both alkaloids significantly reduced CCL-2 expression. The reduction in CCL-2 expression was more significant in sanguisorbin-treated cells, decreasing by 4.3-fold compared to the solvent control group (p<0.001). Two hours after LPS stimulation, compared to the solvent control group, IL-6 mRNA levels were reduced by 3.9-fold (p<0.001) in sanguisorbin-treated cells and by 1.6-fold (p<0.001) in Chelerythrine-treated cells. Compared with Chelerythrine, sanguisorbin was more effective in reducing the gene expression of CCL-2 and IL-6, and its effect was very similar to that of prednisone. Four hours after LPS stimulation, the expression of IL-1RA in cells pretreated with sanguisorbin was significantly higher than that in cells not treated with sanguisorbin (1.7 times higher, p<0.001). Our results help to elucidate the possible mechanisms of action of these alkaloids in the inflammatory process. [2]

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Chelerythrine (CHE) is a quaternary ammonium benzo[c]phenanthridine alkaloid found in plants of the Papaveraceae and Rutaceae families, such as Chelidonium majus and Boletus edulis. [1]
Chelerythrine (CHE) has traditionally been used to treat a variety of inflammatory diseases and has a wide range of biological activities, including antibacterial and anti-inflammatory effects. [1]
In LPS-activated macrophages, the anti-inflammatory mechanism of Chelerythrine (CHE) involves downregulation of the MAPK signaling pathway (particularly inhibition of p38 and ERK1/2 phosphorylation), thereby reducing the production of pro-inflammatory mediators such as TNF-α and NO. [1]
This study suggests that Chelerythrine (CHE) has therapeutic potential for treating inflammation-mediated diseases such as endotoxin shock by regulating macrophage activation. [1]

C21H18NO4.HCL

384.83

383.092

C, 65.71; H, 4.73; Cl, 9.24; N, 3.65; O, 16.67

3895-92-9

Chelerythrine;34316-15-9; 478-03-5 (OH-); 34316-15-9; 3895-92-9 (chloride)

72311

Typically exists as Yellow to orange solids at room temperature

1.36g/cm3

711.4ºC at 760 mmHg

195-205ºC

219.3ºC

1.681

0.72

0

5

2

27

516

0

[Cl-].O1C([H])([H])OC2=C1C([H])=C1C(=C2[H])C([H])=C([H])C2=C3C([H])=C([H])C(=C(C3=C([H])[N+](C([H])([H])[H])=C21)OC([H])([H])[H])OC([H])([H])[H]

WEEFNMFMNMASJY-UHFFFAOYSA-M

InChI=1S/C21H18NO4.ClH/c1-22-10-16-13(6-7-17(23-2)21(16)24-3)14-5-4-12-8-18-19(26-11-25-18)9-15(12)20(14)22;/h4-10H,11H2,1-3H3;1H/q+1;/p-1

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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: ≥ 0.44 mg/mL (1.15 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 4.4 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 2: ≥ 0.44 mg/mL (1.15 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 4.4 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: ≥ 0.43 mg/mL (1.12 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 4.3 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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 (Please use freshly prepared in vivo formulations for optimal results.)