Hepatic epoxide hydrase (increased activity)
Hepatic glutathione-S-transferase (increased activity)
Cytochrome P-450 (reduced level)
Aminopyrine-N-demethylase (reduced activity)
Arylhydrocarbon hydroxylase (AHH) (reduced activity) [2]

In vitro addition of Seneciphylline to rat liver microsomal preparations at concentrations ranging from 10 to 1000 mM caused a slight stimulation of epoxide hydrase activity, with activity increasing from 4.7 ± 0.08 to 5.7 ± 0.09 nmol product formed/min per mg protein at 1000 mM (p < 0.001).
At 10 mM, epoxide hydrase activity was 5.5 ± 0.08 (p < 0.05); at 50 mM, 6.7 ± 0.09 (p < 0.05); at 100 mM, 6.7 ± 0.10 (p < 0.05); at 500 mM, 7.0 ± 0.06 (p < 0.05); at 1000 mM, 7.5 ± 0.03 (p < 0.001). Seneciphylline did not significantly affect glutathione-S-transferase activity in vitro at any concentration tested (10-1000 mM), with values ranging from 238 ± 9 to 257 ± 4 compared to control 238 ± 9.
In vitro, Seneciphylline had no effect on aminopyrine demethylase activity at concentrations from 10 to 1000 mM, with activities ranging from 8.9 ± 0.02 to 9.5 ± 0.06 compared to control 9.5 ± 0.06.
Similarly, Seneciphylline did not affect arylhydrocarbon hydroxylase (AHH) activity in vitro at concentrations from 10 to 1000 mM, with activities ranging from 0.44 ± 0.02 to 0.48 ± 0.01 compared to control 0.44 ± 0.02. [2]

Oral administration of Seneciphylline to male albino rats at a daily dose of 40 mg/kg for 3 days significantly increased hepatic epoxide hydrase activity from 4.6 ± 0.8 to 19.0 ± 1.1 nmol product formed/min per mg protein (p < 0.01). At 80 mg/kg per day for 3 days, epoxide hydrase activity increased further to 24.2 ± 1.4 nmol product formed/min per mg protein (p < 0.01).
Seneciphylline also significantly increased hepatic glutathione-S-transferase activity at 40 mg/kg per day (from 177 ± 6 to 252 ± 8 nmol product formed/min per mg protein, p < 0.01) and at 80 mg/kg per day (240 ± 17, p < 0.05).
Cytochrome P-450 levels were reduced by Seneciphylline from 0.65 ± 0.08 to 0.42 ± 0.07 nmol/mg protein at 40 mg/kg (p < 0.05) and to 0.32 ± 0.06 at 80 mg/kg (p < 0.05).
Aminopyrine demethylase activity decreased from 7.6 ± 0.3 to 6.5 ± 0.3 nmol/min per mg protein at 40 mg/kg (p < 0.05) and to 3.7 ± 0.3 at 80 mg/kg (p < 0.01).
Arylhydrocarbon hydroxylase (AHH) activity decreased from 1.12 ± 0.03 to 0.50 ± 0.08 nmol/min per mg protein at 40 mg/kg (p < 0.01) and to 0.22 ± 0.04 at 80 mg/kg (p < 0.01). [2]

Microsomal epoxide hydrase activity was measured using [14C]styrene oxide as substrate. Rat liver microsomes were incubated with [14C]styrene oxide, and the formation of styrene glycol was quantified radiometrically. For in vivo studies, livers were removed from treated animals, homogenized in 1.15% KCl with 0.02 M HEPES (pH 7.4), centrifuged at 10,000 × g for 20 min, and the supernatant further centrifuged at 105,000 × g for 1 hour to obtain microsomal pellets. The microsomal pellet was washed and resuspended in HEPES buffer. For in vitro studies, various concentrations of Seneciphylline (10-1000 mM) were added directly to the incubation mixture containing control liver microsomes. [2]
Glutathione-S-transferase activity was determined radiometrically using [8-14C]styrene oxide as substrate in the cytosol fraction (105,000 × g supernatant). The formation of glutathione conjugates was measured. For in vivo studies, cytosol was prepared from livers of treated rats. For in vitro studies, Seneciphylline at concentrations of 10-1000 mM was added to the incubation mixture. [2]
Cytochrome P-450 content was analyzed by the method of Omura and Sato, based on the carbon monoxide difference spectrum of reduced microsomes. [2]
Microsomal aminopyrine-N-demethylase activity was assayed by measuring the amount of formaldehyde formed using the Nash colorimetric method, with a final substrate concentration of 12 mM aminopyrine. [2]
Arylhydrocarbon hydroxylase (AHH) activity was determined using benzo[a]pyrene as substrate (final concentration 0.6 mM) by the method of Nebert and Gelboin, measuring the formation of fluorescent phenolic products. [2]

Male Swiss albino rats weighing 155-175 g were used. Seneciphylline was first dissolved in 0.2 N HCl, neutralized with NaOH, and brought to volume with normal saline. The alkaloid was administered orally (p.o.) once daily for 3 consecutive days at doses of 40 mg/kg or 80 mg/kg body weight per day. Control animals received normal saline only. Animals were killed 24 hours after the last dose. Livers were removed, weighed, and processed for microsomal and cytosolic fractions. [2]

Absorption, Distribution and Excretion
Animal studies have shown that the highest concentrations are found in the liver, lungs, kidneys, and spleen. Following injection, the radioactive material is rapidly excreted in urine and feces within 16 hours (84% or more). More than 1.5% of the dose remains in the liver after 16 hours. A small amount (0.04%) of the dose is transferred to milk within 16 hours; most of the radioactive material is present in skim milk, indicating that the pyrrolizidine alkaloids are transferred to milk as water-soluble metabolites. …The binding to calf thymus DNA and microsomal macromolecules was determined in vitro. Binding was weakened in the absence of oxygen or NADPH-generating systems, or by boiling the microsomes. No inhibitory effect of potassium cyanide on binding was observed. Pyrrolizidine alkaloids…To investigate its transfer to milk, a cow was orally administered a single dose of 1 mg/kg body weight of (3H) senna. The presence of radioactive substances from this compound in blood and milk was subsequently monitored. Based on senglitin, the blood concentration exceeded 100 ng/mL within the first 18 hours after administration. At 54 hours, the blood concentration remained at 11 ng/mL. Similar alkaloid levels were observed in milk. At 64 hours, the milk concentration was still 5 ng/mL. A total of 0.16% of the dose was excreted through milk. Three weeks after administration, 40 ng/g of alkaloids (0.06% of the administered dose) were detected in the liver. In addition to unchanged senglitin and retroline, N-oxides were also detected in milk casserotonin metabolites (11.2% at 27 hours). Metabolites/Metabolites Typically, hepatotoxic pyrrolizidine alkaloids are metabolized in the rat liver to form hydrolysates, N-oxides, and dehydropyrrolizidine (pyrrole) derivatives. ...Dehydroalkaloids are highly reactive alkylating agents... /Pyrrolizidine alkaloids/
Dehydroretronine...water-soluble pyrrole metabolites of senna...have been shown to be carcinogenic. /Pyrrolizidine alkaloids/
In animals, the main metabolic pathways of pyrrolizidine alkaloids are: (a) ester hydrolysis; (b) N-oxidation; (c) nuclear dehydrogenation of pyrrolizidine to form pyrrole derivatives. Pathways (a) and (b) are considered detoxification mechanisms. Pathway (c) produces toxic metabolites. Pathway (a) occurs in the liver and blood; pathways (b) and (c) are mediated by the hepatic microsomal mixed-function oxidase system. /Pyrrolizidine alkaloids/

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Biological half-life
Only a relatively small proportion of the administered dose remains in the body within a few hours. The majority of this remains as metabolites bound to tissue components. Following intravenous injection in animals, pyrrolizidine N-oxide disappears from the serum, with an initial half-life of 3-20 minutes. /Pyrrolizidine alkaloids/

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Non-Human Toxicity Values
Mouse intravenous LD50: 90 mg/kg Rat intravenous LD50: 80 mg/kg Male rat intraperitoneal LD50: 77 mg/kg (95% confidence interval: 71-86 mg/kg) Female rat intraperitoneal LD50: 83 mg/kg (95% confidence interval: 77-90 mg/kg)

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Oral administration of Seneciphylline at 40 and 80 mg/kg per day for 3 days caused significant reduction in hepatic cytochrome P-450 levels and monooxygenase activities (aminopyrine demethylase and AHH), indicating hepatotoxic effects on drug metabolizing enzymes. The compound is a pyrrolizidine alkaloid known to be metabolized by hepatic monooxygenases to N-oxide and pyrrole derivatives, which are strong alkylating agents that can bind to nucleophilic sites of cellular macromolecules. Seneciphylline may inhibit protein synthesis, as pyrrolizidine alkaloids have been shown to impair synthesis of monooxygenase enzymes and cytochrome P-450. The reduction in enzyme activities could also be due to inactivation of cytochrome P-450 by reactive pyrrole metabolites. [2]

[1]. Cytotoxicity of Senecio in macrophages is mediated via its induction of oxidative stress. Res Vet Sci. 2009 Aug;87(1):85-90.

[2]. Effect of seneciphylline and senecionine on hepatic drug metabolizing enzymes in rats. J Ethnopharmacol. 1984 Dec;12(3):271-8.

Senecio alkaloid is a white powder. (NTP, 1992)
LSM-2853 is a citrate compound.
Sensio alkaloid has been reported to exist in Senecio carniolica, Senecio rodriguezii, and other organisms with relevant data.
Mechanism of Action

Mixed-function oxidases activate the alkaloid to produce pyrrole dehydroalkaloids, which are active alkylating agents. Metabolites bind to hepatocytes, leading to liver necrosis. Some metabolites are released into the bloodstream and are believed to reach the lungs via the liver, causing vascular damage. Pyrrole metabolites are cytotoxic, acting on hepatocytes and vascular endothelial cells in the liver and lungs. Pyrrolizidine Alkaloids
This study investigated the effects of oral administration of the pyrrolizidine alkaloid, Senecio scandens (from the plant Senecio scandens), on the activities of epoxide hydrolase, glutathione S-transferase, aminopyrine N-demethylase, and aryl hydrocarbon hydroxylase (AHH) in the liver microsomes of young male albino rats. The results showed that Senecio scandens significantly increased the activities of epoxide hydrolase and glutathione S-transferase, but decreased the activities of cytochrome P-450 and its associated monooxygenases. …Senecio scandens…had no significant in vitro effects on the studied hepatic drug-metabolizing enzymes; both alkaloids slightly stimulated the activity of epoxide hydrolase, while Senecio scandens slightly decreased the activity of aminopyrine demethylase.

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Seneciphylline is one of the pyrrolizidine alkaloids found in Senecio vulgaris (Compositae), a plant used in traditional medicine in India for dysmenorrhoea, amenorrhoea, as a diaphoretic, diuretic, tonic, emmenagogue, and for chronic mastitis, haemorrhoids, gout, helminthiasis, and as a purgative and emetic. The present investigation suggests that medicinal use of Senecio vulgaris is doubtful due to the significant effects of its alkaloids on hepatic drug metabolizing enzymes. [2]
The chemical structure of Seneciphylline differs from senecionine in stereochemistry: at position 8, the H atom is in β-position (vs. α in senecionine); at position 12, the CH3 group has β-configuration (vs. α in senecionine); and position 13 is saturated in seneciphylline whereas it is unsaturated in senecionine. These differences may account for the distinct effects on epoxide hydrase and glutathione-S-transferase. [2]

C18H23NO5

333.3789

333.157

480-81-9

5281750

White to off-white solid

1.3±0.1 g/cm3

577.7±50.0 °C at 760 mmHg

217ºC

303.2±30.1 °C

0.0±3.6 mmHg at 25°C

1.581

0.81

1

6

0

24

650

3

O1C(/C(=C(/[H])\C([H])([H])[H])/C([H])([H])C(=C([H])[H])[C@](C([H])([H])[H])(C(=O)OC([H])([H])C2=C([H])C([H])([H])N3C([H])([H])C([H])([H])[C@]1([H])[C@]32[H])O[H])=O

FCEVNJIUIMLVML-QPSVUOIXSA-N

InChI=1S/C18H23NO5/c1-4-12-9-11(2)18(3,22)17(21)23-10-13-5-7-19-8-6-14(15(13)19)24-16(12)20/h4-5,14-15,22H,2,6-10H2,1,3H3/b12-4-/t14-,15-,18-/m1/s1

(1R,4Z,7R,17R)-4-ethylidene-7-hydroxy-7-methyl-6-methylidene-2,9-dioxa-14-azatricyclo[9.5.1.014,17]heptadec-11-ene-3,8-dione

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 : ~7.69 mg/mL (~23.07 mM)

Solubility in Formulation 1: ≥ 0.77 mg/mL (2.31 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 7.7 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: ≥ 0.77 mg/mL (2.31 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 7.7 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: ≥ 0.77 mg/mL (2.31 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 7.7 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.)