Stable isotope‐labelled morphine to study <i>in vivo</i> central and peripheral morphine glucuronidation and brain transport in tolerant mice

Weinsanto, Ivan; Laux-Biehlmann, Alexis; Mouheiche, Jinane; Maduna, Tando; Delalande, François; Chavant, Virginie; Gabel, Florian; Darbon, Pascal; Charlet, Alexandre; Poisbeau, Pierrick; Lamshöft, Marc; Dorsselaer, Alain Van; Cianférani, Sarah; Parat, Marie-Odile; Goumon, Yannick

doi:10.1111/bph.14454

Cited by 9 publications

(7 citation statements)

References 58 publications

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“…Then, morphine is taken up by glial cells including microglial cells and astrocytes through the action of the organic cation transporter OCT1 (Tzvetkov et al, 2013). After conversion by UDP-glucuronosyl-transferase (UGT) enzymes (King et al, 2000; Weinsanto et al, 2018), glucuronides are released through the efflux MRP3 transporter (multidrug resistance protein 3; Zelcer et al, 2005). In this context, it is plausible that after administration and cellular uptake, morphine can form complexes with cytoplasmic CK-B and therefore negatively modulate its activity.…”

Section: Discussionmentioning

confidence: 99%

Morphine Binds Creatine Kinase B and Inhibits Its Activity

Weinsanto

Mouheiche

Laux-Biehlmann

et al. 2018

Front. Cell. Neurosci.

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Morphine is an analgesic alkaloid used to relieve severe pain, and irreversible binding of morphine to specific unknown proteins has been previously observed. In the brain, changes in the expression of energy metabolism enzymes contribute to behavioral abnormalities during chronic morphine treatment. Creatine kinase B (CK-B) is a key enzyme involved in brain energy metabolism. CK-B also corresponds to the imidazoline-binding protein I2 which binds dopamine (a precursor of morphine biosynthesis) irreversibly. Using biochemical approaches, we show that recombinant mouse CK-B possesses a μM affinity for morphine and binds to morphine in vitro. The complex formed by CK-B and morphine is resistant to detergents, reducing agents, heat treatment and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). CK-B-derived peptides CK-B1–75 and CK-B184–258 were identified as two specific morphine binding-peptides. In vitro, morphine (1–100 μM) significantly reduces recombinant CK-B enzymatic activity. Accordingly, in vivo morphine administration (7.5 mg/kg, i.p.) to mice significantly decreased brain extract CK-B activity compared to saline-treated animals. Together, these results show that morphine strongly binds CK-B and inhibits its activity in vitro and in vivo.

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Section: Discussionmentioning

confidence: 99%

Morphine Binds Creatine Kinase B and Inhibits Its Activity

Weinsanto

Mouheiche

Laux-Biehlmann

et al. 2018

Front. Cell. Neurosci.

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“…Therefore, our main hypothesis is that the behavioral differences in the analgesic effect of morphine between male and female mice rely on the central metabolism of morphine. Indeed, several in vitro studies have shown the capability of brain homogenates and glial cells to metabolize morphine into M3G in both mice and humans (13, 37). In addition, even though M3G displays low BBB permeability (38), we showed here that higher levels of M3G were present in different brain regions following an i.p.…”

Section: Discussionmentioning

confidence: 99%

“…Morphine analgesic tolerance resulting from chronic treatment corresponds to the loss of morphine efficacy and the need for higher doses to achieve sufficient analgesia (13). Although several mechanisms implicating MORs have been previously described to explain this phenomenon (for review, see (14)), neuroinflammatory processes have been proposed to be involved in tolerance mechanisms (15).…”

Section: Introductionmentioning

confidence: 99%

Central metabolism as a potential origin of sex differences in morphine analgesia but not in the induction of analgesic tolerance in mice

Gabel

Hovhannisyan

Andry

et al. 2020

Preprint

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In rodents, morphine analgesia is influenced by sex. However, conflicting results exist regarding the interaction between sex and morphine analgesic tolerance. Morphine is metabolized in the liver and brain into morphine-3-glucuronide (M3G). Sex differences in morphine metabolism and differential metabolic adaptations during tolerance development might explain the behavioral discrepancies. The present article investigates the differences in peripheral and central morphine metabolism after acute and chronic morphine treatment in male and female mice.The first experiment aimed to determine whether morphine analgesia and tolerance differ between male and female mice using the tail-immersion test. The second experiment evaluated morphine and M3G metabolic kinetics in the blood using LC-MS/MS. Morphine and M3G were also quantified in several central nervous system (CNS) regions after acute and chronic morphine treatment. Finally, the blood-brain barrier permeability of M3G was assessed in male and female mice.This study demonstrated that female mice showed weaker morphine analgesia. In addition, tolerance appeared earlier in females but the sex discrepancies observed seemed to be due to the initial differences in morphine analgesia rather than to sex-specific mechanisms involving metabolism. Additionally, compared to male mice, female mice showed higher levels of M3G in the blood and in several CNS regions, whereas lower levels of morphine were observed in these brain regions. These differences are attributable mainly to morphine central metabolism, which differed between males and females in pain-related brain regions, consistent with the weaker analgesic effect in females. However, the role of morphine metabolism in analgesic tolerance seems rather limited.

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“…As 4OH-tamoxifen is the major active metabolite of tamoxifen in mice, in vitro experiments were performed on mouse liver microsomes to study the impact of 500 µM of morphine on the glucuronidation of 4OH-tamoxifen. Morphine was used at 500 µM to determine the K m of the glucuronidation of 4OH-tamoxifen as this concentration corresponds to the K m previously determined for morphine glucuronidation in mice (12,19). As shown in Figure 3, morphine significantly affects the production of 4OH-tamoxifen-glucuronide.…”

Section: Enzymatic Study In Vitromentioning

confidence: 96%

“…Its metabolism in the Abbreviations: ACN, acetonitrile; ADME, absorption, distribution, metabolism, and/or excretion; AF, formic acid; AI, aromatase-inhibitors; CYP, cytochrome P450; ER, estrogen receptor; i.p., injected intraperitoneally; IS, internal standard; LC-MS/MS, liquid chromatography-mass spectrometry/mass spectrometry; M3G, morphine-3-glucuronide; M6G, morphine-6-glucuronide; MOR, Mu opioid receptors; SEM, standard error of the mean; SERM, selective estrogen receptor modulator; TDM, therapeutic drug monitoring; UGT, UDP-glucuronosyltransferases; SULT, sulfotransferase. liver and brain leads mainly to the formation of morphine-3glucuronide (M3G) (10) and morphine-6-glucuronide (M6G) (10)(11)(12). In human, morphine-glucuronidation is catalyzed by UGT2B7 and to a lower extent by a number of other UGT isoforms (UGT1A10, UGT1A1, 1A3, 1A6, 1A8, 1A9, 2A1, and UGT2B21) (9,13,14).…”

Section: Introductionmentioning

confidence: 99%

Unveiling the Impact of Morphine on Tamoxifen Metabolism in Mice in vivo

et al. 2020

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Background: Tamoxifen is used to treat breast cancer and cancer recurrences. After administration, tamoxifen is converted into two more potent antitumor compounds, 4OH-tamoxifen and endoxifen by the CYP3A4/5 and 2D6 enzymes in human. These active compounds are inactivated by the same UDP-glucuronosyltransferase isoforms as those involved in the metabolism of morphine. Importantly, cancer-associated pain can be treated with morphine, and the common metabolic pathway of morphine and tamoxifen suggests potential clinically relevant interactions. Methods: Mouse liver microsomes were used to determine the impact of morphine on 4OH-tamoxifen metabolism in vitro. For in vivo experiments, female mice were first injected with tamoxifen alone and then with tamoxifen and morphine. Blood was collected, and LC-MS/MS was used to quantify tamoxifen, 4OH-tamoxifen, N-desmethyltamoxifen, endoxifen, 4OH-tamoxifen-glucuronide, and endoxifen-glucuronide. Results: In vitro, we found increased K m values for the production of 4OH-tamoxifen-glucuronide in the presence of morphine, suggesting an inhibitory effect on 4OH-tamoxifen glucuronidation. Conversely, in vivo morphine treatment decreased 4OH-tamoxifen levels in the blood while dramatically increasing the formation of inactive metabolites 4OH-tamoxifen-glucuronide and endoxifen-glucuronide. Conclusions: Our findings emphasize the need for caution when extrapolating results from in vitro metabolic assays to in vivo drug metabolism interactions. Importantly, morphine strongly impacts tamoxifen metabolism in mice. It suggests that tamoxifen efficiency could be reduced when both drugs are co-administered in a clinical setting, e.g., to relieve pain in breast cancer patients. Further studies are needed to assess the potential for tamoxifen-morphine metabolic interactions in humans.

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Stable isotope‐labelled morphine to study in vivo central and peripheral morphine glucuronidation and brain transport in tolerant mice

Abstract: Our data suggests that tolerance to the analgesic effects of morphine is not linked to increased glucuronidation to M3G or to altered global BBB permeability of morphine.

Cited by 9 publications

References 58 publications

Morphine Binds Creatine Kinase B and Inhibits Its Activity

Morphine Binds Creatine Kinase B and Inhibits Its Activity

Central metabolism as a potential origin of sex differences in morphine analgesia but not in the induction of analgesic tolerance in mice

Unveiling the Impact of Morphine on Tamoxifen Metabolism in Mice in vivo

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