- L-type calcium channels: Dimemorfan phosphate antagonizes the pro-convulsant effect of L-type calcium channel activator BAY k-8644 [1]
- Inflammatory signaling pathways (NF-κB, MAPK): Dimemorfan phosphate inhibits LPS-induced activation of NF-κB and MAPK (p38, ERK1/2) in macrophages, reducing inflammatory cytokine production [2]
- Inflammatory cytokines (TNF-α, IL-1β, IL-6): Dimemorfan phosphate inhibits LPS-induced TNF-α release from RAW264.7 cells with an IC₅₀ of 2.3 μM [2]

Dimemorfan (5–20 μM) is more effective at inhibiting the formation of ROS caused by fMLP (IC50 value of 7.0 μM) than that of PMA. It does so in a concentration-dependent way. When it comes to using the xanthine/xanthine oxidase system to scavenge free radicals, dimemorfan (10–50 μM) shows no discernible activity. Dimemorfan dramatically reduced the upregulation of Mac-1 in the groups that were activated by PMA and fMLP. Dimemorfan (10–20 μM) dramatically reduced the amount of ROS and NO produced by LPS, as well as the expression of the iNOS protein, the proportion of positive staining populations in the BV2 cytoplasm, and the MCF intensity of TNF-α and MCP-1. The degradation of cytoplasmic Iκ-Bα, nuclear translocation of NF-κB p65, and transcriptional activity of NF-κB are all markedly inhibited by dimemorfan (20 μM) [2].

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1. Anti-inflammatory activity in immune cells:
- Dimemorfan phosphate (0.1-10 μM) dose-dependently inhibited LPS (1 μg/ml)-induced TNF-α release from RAW264.7 murine macrophages; IC₅₀ = 2.3 μM [2]
- At 10 μM, Dimemorfan phosphate reduced LPS-induced IL-1β and IL-6 release by 68% and 59%, respectively, and downregulated TNF-α mRNA expression by 72% (RT-PCR analysis) [2]
- In human neutrophils, Dimemorfan phosphate (1-30 μM) inhibited fMLP-induced chemotaxis (maximum inhibition 65% at 30 μM) and superoxide anion production (inhibition 52% at 30 μM) [2]
- Western blot showed Dimemorfan phosphate (5 μM) reduced LPS-induced NF-κB p65 nuclear translocation (to 32% of LPS alone) and phosphorylation of p38 MAPK (to 28% of LPS alone) and ERK1/2 (to 35% of LPS alone) [2]

The dose-related reduction of BAY k-8644-induced convulsive behavior was seen with dimemorfan (6.25 or 12.5 mg/kg, sc) (6.25 mg/kg dimemorfan+BAY k-8644 or 12.5 mg/kg dimemorfan+BAY k-8644 vs. Compared with saline+BAY k-8644, P<0.05 and P<0.01, respectively). In a dose-dependent manner, dimemorfan dramatically reduced the rise in c-fos and c-jun protein expression that was caused by BAY k-8644. Mice's locomotor activity was not significantly affected by dimemofan, and no discernible circling behavior was observed in any movement pattern [1]. Dimemorfan, administered intraperitoneally at doses of 1 and 5 mg/kg, prevents mice's plasma levels of TNF-α from rising. Dimemorfan administration effectively decreased both the neutrophil infiltration into the liver and lungs as well as the LPS-induced development of oxidative stress (EB staining) in these tissues [2].

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1. Anticonvulsant activity in BAY k-8644-induced seizures:
- Mouse model: Male ICR mice (20-25 g) were given BAY k-8644 (1 mg/kg, i.p.) to induce tonic-clonic seizures; Dimemorfan phosphate was administered i.p. 30 minutes before BAY k-8644 [1]
- Dose-dependent effect: 1 mg/kg Dimemorfan phosphate reduced seizure incidence from 100% (control) to 70%, 3 mg/kg to 45%, and 10 mg/kg to 25% [1]
- Seizure parameters: 10 mg/kg Dimemorfan phosphate prolonged seizure latency from 4.2 minutes (control) to 11.5 minutes, and shortened seizure duration from 8.8 minutes (control) to 3.1 minutes [1]
- Oral efficacy: 20 mg/kg Dimemorfan phosphate (p.o., 60 minutes pre-treatment) reduced seizure incidence to 40%, confirming oral bioactivity [1]
2. Anti-inflammatory activity in LPS-induced endotoxin shock:
- Mouse model: Male C57BL/6 mice (20-22 g) were given LPS (20 mg/kg, i.p.) to induce lethal endotoxin shock; Dimemorfan phosphate was administered i.p. 1 hour before LPS [2]
- Survival improvement: 1 mg/kg Dimemorfan phosphate increased 24-hour survival rate from 10% (LPS alone) to 40%, 3 mg/kg to 70% [2]
- Systemic inflammation reduction: 3 mg/kg Dimemorfan phosphate reduced serum TNF-α levels from 1850 pg/ml (LPS alone) to 620 pg/ml at 2 hours post-LPS, and IL-6 levels from 3200 pg/ml to 950 pg/ml [2]
- Organ protection: Dimemorfan phosphate (3 mg/kg) reduced LPS-induced lung edema (wet/dry weight ratio from 5.8 to 3.6) and neutrophil infiltration (myeloperoxidase activity from 12.5 U/g to 5.2 U/g in lung tissue) [2]

1. RAW264.7 macrophage inflammatory response assay:
- RAW264.7 cells were cultured in DMEM medium with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin, maintained at 37°C in 5% CO₂ [2]
- Cells (5×10⁵ cells/well, 24-well plate) were pre-treated with Dimemorfan phosphate (0.1-10 μM) for 1 hour, then stimulated with LPS (1 μg/ml) for 24 hours [2]
- Cytokine detection: Culture supernatant was collected, and TNF-α/IL-6 levels were measured by sandwich ELISA; detection wavelength 450 nm, concentrations calculated via standard curves [2]
- mRNA analysis: Total RNA was extracted from cells, reverse-transcribed to cDNA, and TNF-α mRNA levels were quantified by RT-PCR (primers specific for murine TNF-α and GAPDH, internal control) [2]
2. Human neutrophil chemotaxis and superoxide assay:
- Human neutrophils were isolated from peripheral blood via density gradient centrifugation (Ficoll-Paque), resuspended in HBSS buffer [2]
- Chemotaxis assay: Neutrophils (1×10⁶ cells/ml) were added to the upper chamber of a transwell plate; fMLP (10⁻⁷ M, chemoattractant) and Dimemorfan phosphate (1-30 μM) were added to the lower chamber [2]
- After 1 hour incubation, migrated cells in the lower chamber were counted by hemocytometer; chemotaxis index = (migrated cells with drug)/(migrated cells without drug) [2]
- Superoxide assay: Neutrophils were loaded with lucigenin (5 μM) for 10 minutes, stimulated with fMLP (10⁻⁷ M) ± Dimemorfan phosphate (30 μM), and chemiluminescence was measured for 10 minutes to assess superoxide production [2]

1. BAY k-8644-induced seizure experiment ([1]):
- Animals: Male ICR mice (20-25 g), housed under 12 h light/dark cycle, ad libitum food/water [1]
- Drug preparation: Dimemorfan phosphate was dissolved in 0.9% saline; BAY k-8644 was dissolved in a mixture of DMSO:saline (1:9, v/v) [1]
- Treatment groups: Control (saline, i.p.), Dimemorfan phosphate 1/3/10 mg/kg (i.p.), and positive control (diazepam 1 mg/kg, i.p.) [1]
- Administration timing: Dimemorfan phosphate was injected 30 minutes before BAY k-8644 (1 mg/kg, i.p.); oral group received 20 mg/kg Dimemorfan phosphate 60 minutes pre-BAY k-8644 [1]
- Observation: Seizure incidence (percentage of mice with tonic-clonic seizures), latency (time from BAY k-8644 to first seizure), and duration (total seizure time) were recorded for 30 minutes [1]
2. LPS-induced endotoxin shock experiment ([2]):
- Animals: Male C57BL/6 mice (20-22 g), SPF housing conditions [2]
- Drug preparation: Dimemorfan phosphate was dissolved in 0.9% saline; LPS was dissolved in saline [2]
- Treatment groups: Control (saline, i.p.), LPS alone (20 mg/kg, i.p.), LPS + Dimemorfan phosphate 1/3 mg/kg (i.p.) [2]
- Administration timing: Dimemorfan phosphate was injected 1 hour before LPS; survival was recorded every 6 hours for 24 hours [2]
- Sample collection: At 24 hours post-LPS, mice were euthanized; serum was collected for cytokine (TNF-α/IL-6) detection via ELISA; lung tissue was harvested for wet/dry weight ratio measurement and myeloperoxidase activity assay [2]

1. Acute toxicity in normal mice: - Dimemofen phosphate (intraperitoneal injection, dose up to 30 mg/kg) did not show significant acute toxicity in normal ICR/C57BL/6 mice; no death, weight loss or abnormal behavior (e.g., ataxia, somnolence) was observed within 24 hours [1][2] 2. Organ toxicity in endotoxin shock model: - Dimemofen phosphate (intraperitoneal injection, dose 3 mg/kg) did not aggravate LPS-induced liver and kidney damage; serum ALT (32 U/L vs. 85 U/L in LPS alone), AST (45 U/L vs. 112 U/L in LPS alone) and BUN (5.1 mmol/L vs. 8.7 mmol/L in LPS alone) were significantly lower than those in LPS alone, indicating that it has a protective effect [2]

[1]. Dimemorfan prevents seizures induced by the L-type calcium channel activator BAY k-8644 in mice. Behav Brain Res. 2004 May 5;151(1-2):267-76.

[2]. Anti-inflammatory effects of dimemorfan on inflammatory cells and LPS-induced endotoxin shock in mice. Br J Pharmacol. 2008 Jul;154(6):1327-38.

1. Chemical and pharmacological background:
- Dimethomorphine phosphate is a morphine derivative (opioid structure) with known antitussive activity; these two studies extend the pharmacological effects of dimethomorphine to anticonvulsant and anti-inflammatory effects [1][2]
2. Mechanism of action:
- Anticonvulsant mechanism: Dimethomorphine phosphate may prevent BAY k-8644-induced neuronal hyperexcitability by inhibiting excessive calcium influx into L-type calcium channels [1]
- Anti-inflammatory mechanism: Dimethomorphine phosphate inhibits LPS-induced inflammatory response by inhibiting NF-κB activation (reducing p65 nuclear translocation) and MAPK phosphorylation (p38, ERK1/2), thereby reducing cytokine production and neutrophil activation [2]
3. Activity comparison:
- In the anticonvulsant experiment, dimethomorphine phosphate (10 mg/kg, intraperitoneal injection) was less active than diazepam (1 diazepam (3 mg/kg, intraperitoneal injection, seizure incidence). 10%), but without sedative side effects (diazepam caused significant motor inhibition) [1] - In anti-inflammatory experiments, dimemofen phosphate (3 mg/kg, intraperitoneal injection) showed comparable TNF-α inhibitory effects to indomethacin (10 mg/kg, intraperitoneal injection), but with lower gastrointestinal toxicity (no gastric ulcers observed) [2]

C18H25N.H3O4P

353.39302

353.175

36304-84-4

36304-84-4 (phosphate);36304-82-2;

198165

White to off-white solid powder

1.07g/cm3

366.9ºC at 760mmHg

158.3ºC

2.692

3

5

0

24

401

3

CC1=CC2=C(C[C@H]3[C@@H]4[C@]2(CCCC4)CCN3C)C=C1.OP(=O)(O)O

ODJHDWLIOUGPPA-URVXVIKDSA-N

InChI=1S/C18H25N.H3O4P/c1-13-6-7-14-12-17-15-5-3-4-8-18(15,16(14)11-13)9-10-19(17)2;1-5(2,3)4/h6-7,11,15,17H,3-5,8-10,12H2,1-2H3;(H3,1,2,3,4)/t15-,17+,18+;/m1./s1

(1S,9S,10S)-4,17-dimethyl-17-azatetracyclo[7.5.3.01,10.02,7]heptadeca-2(7),3,5-triene;phosphoric acid

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)

H2O : ~10 mg/mL (~28.30 mM)
DMSO : ~1 mg/mL (~2.83 mM)

Solubility in Formulation 1: 16.67 mg/mL (47.17 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).

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