A Novel Antifolate, MX-68, Inhibits the Development of Autoimmune Disease in MRL/lpr Mice

Mihara, Masahiko; Takagi, Nobuyuki; Urakawa, Kazumi; Moriya, Yoichiro; Takeda, Yasuhisa

doi:10.1159/000237622

Cited by 7 publications

(2 citation statements)

References 14 publications

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“…Like methotrexate, MX-68 has a high affinity for the enzyme dihydrofolate reductase and inhibits the proliferation of human peripheral blood mononuclear cells, endothelial cells, and synovial fibroblasts in vitro (5). In vivo, MX-68 prevents collageninduced arthritis in mice (5) and rats (6), adjuvantinduced arthritis in rats (4), and autoimmune nephritis in lupus mice (7,8). Furthermore, both MX-68 and methotrexate increased the release of adenosine from Daudi cells, an effect that is mediated by inhibition of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) transformylase, and inhibited the accumulation of leukocytes into the murine air pouch after carrageenan injection, an effect that was abolished by injection of the adenosine A 2 receptor antagonist 3,7-dimethyl-1-propargylxanthine (9).…”

Section: Mx-68 (N-[[4-[(24-diaminopteridin-6-yl)methyl]-34-dihydro-mentioning

confidence: 99%

Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Montesinos¹,

Desai²,

Delano³

et al. 2003

Arthritis & Rheumatism

189

116

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Objective. Low-dose weekly methotrexate therapy remains a mainstay in the treatment of inflammatory arthritis. Results of previous studies demonstrated that adenosine, acting at one or more of its receptors, mediates the antiinflammatory effects of methotrexate in animal models of both acute and chronic inflammation. We therefore sought to establish which receptor ( Results. Low-dose weekly methotrexate treatment increased the adenosine concentration in the exudates of all mice studied and reduced leukocyte and tumor necrosis factor ␣ accumulation in the exudates of wildtype mice, but not in those of A 2A or A 3 receptor knockout mice. Dexamethasone, an agent that suppresses inflammation by a different mechanism, was equally effective at suppressing leukocyte accumulation in A 2A knockout, A 3 knockout, and wild-type mice, indicating that the lack of response was specific for methotrexate and MX-68.Conclusion. These findings confirm that adenosine, acting at A 2A and A 3 receptors, is a potent regulator of inflammation. Moreover, these results provide strong evidence that adenosine, acting at either or both of these receptors, mediates the antiinflammatory effects of methotrexate and its analog MX-68.Low-dose weekly methotrexate is the "gold standard" of therapy in rheumatoid arthritis and other inflammatory diseases. Methotrexate's mechanism of action in the treatment of inflammatory diseases has been the subject of some controversy, although in previous studies, investigators in our group have demonstrated that adenosine mediates the antiinflammatory effects of methotrexate treatment in models of acute and chronic inflammation (1,2). Adenosine, whether released from injured cells or tissues or applied exogenously, regulates inflammation via interaction with one or more of the 4 known receptors for adenosine (A 1 , A 2A , A 2B , and A 3 ), as demonstrated by many in vitro and in vivo pharmacologic studies (for review, see ref.3). The demonstration that adenosine mediates the antiinflammatory effects of methotrexate in in vivo models of acute inflammation rests upon reversal of the antiinflammatory effects of methotrexate, either by enzymatic hydrolysis of adenosine by adenosine deaminase or by administration of adenosine receptor antagonists to reverse the antiinflammatory effects of methotrexate treatment (1,2). Although the antiinflammatory effects of methotrexate are mediated by multiple adenosine receptors in the adjuvant arthritis model of inflammation (2), the identity of the receptor(s) involved in the suppression of inflammation in models of acute inflammation has not been so well characterized.

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Section: Mx-68 (N-[[4-[(24-diaminopteridin-6-yl)methyl]-34-dihydro-mentioning

confidence: 99%

Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Montesinos¹,

Desai²,

Delano³

et al. 2003

Arthritis & Rheumatism

189

116

View full text Add to dashboard Cite

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“…Recently, several reports have appeared on the usefulness of MX-68 and describe the inhibition of the development of collagen-induced arthritis (CIA) in mice (Mihara et al 1996) and autoimmune kidney disease in lupus mice (Mihara et al 1997a). Moreover, MX-68 suppresses immunoglobulin production in-vivo and in-vitro (Mihara et al 1997b).…”

Section: Introductionmentioning

confidence: 99%

Effect of MX-68 on airway inflammation and hyperresponsiveness in mice and guinea-pigs

Nagao

Akabane

Masuda

et al. 2004

Journal of Pharmacy and Pharmacology

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MX-68 is a newly synthesized antifolate, which is a derivative of methotrexate (MTX). In this paper, the effect of MX-68 on allergic airway responses in mice and guinea-pigs was studied. In the first experiment, antigen-induced airway inflammation and airway hyperresponsiveness (AHR) to acetylcholine in mice were examined and compared with the effects of classical antifolate methotrexate and prednisolone. Mice were sensitized with ovalbumin as an antigen and challenged with ovalbumin inhalation three times. After the last inhalation, AHR and airway inflammation were observed. An increase in Th2 cytokines (IL-4 and IL-5) and a decrease in a Th1 cytokine (IFN-gamma) in the bronchoalveolar lavage fluid (BALF), as well as an elevation of the immunoglobulin level in serum, were observed in sensitized mice. Oral administration of MX-68 had no effect on changes of body weight, but prednisolone reduced body weight during the experiment. The antigen-induced AHR and increases in the number of eosinophils and lymphocytes in BALF were significantly inhibited by MX-68. MX-68 interfered with the elevation of IL-4 and IL-5 levels in BALF, but had no effect on the decrease in IFN-gamma. Moreover, MX-68 significantly inhibited the elevation of serum IgE and IgG levels. In the guinea-pig model for bronchial asthma, biphasic increases in airway resistance (immediate asthmatic response, IAR, and late asthmatic response, LAR), as well as accumulated inflammatory cells in BALF, were observed after repeated antigen challenge. These asthmatic responses and inflammatory signs were significantly decreased by administration of MX-68. These results suggest that MX-68 obviously has an anti-inflammatory effect in an animal model of asthma and would be useful clinically for the treatment of bronchial asthma.

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Binding and transport of methotrexate and its derivative, MX-68, across the brush-border membrane in rat kidney

Han

Kato

Sugiyama

1999

Biopharm. Drug Dispos.

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Binding and transport properties of methotrexate (MTX) and its novel derivative, MX-68, were examined in brush-border membrane vesicles (BBMVs) isolated from rat kidneys. The uptake of MTX, MX-68 and folic acid by BBMVs was stimulated by an inwardly-directed H(+) gradient. Such H(+)-dependent uptake of folic acid is compatible with a previous report (Bhandari et al., Biochim Biophys Acta 1988; 937: 211). The MTX uptake exhibits saturation with a K(m) of 0.834 microM. Although the uptake of these three compounds at optimal pH depended on the osmolarity of the medium, a substantial portion of the uptake was osmolarity-insensitive. By changing the medium osmolarity, the uptake by BBMVs could be separately discriminated as osmolarity-sensitive and insensitive portions, representing transport into the intravesicular space and binding to the surface of BBMVs, respectively. For all three compounds, the binding increased in a time-dependent manner, while the amount transported reached a maximum after a relatively short incubation period. The transport of folic acid, but not its binding, exhibited an overshoot phenomenon under conditions of an inward H(+) gradient. The present results suggest that reabsorption of MTX and MX-68 in the kidney is governed by both their binding and transport mechanisms, with a similar kinetic profile to that of folic acid. The involvement of a transport system seems to make a relatively small contribution to the reabsorption of MTX assessed in BBMVs compared with MX-68 and folic acid.

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A Novel Antifolate, MX-68, Inhibits the Development of Autoimmune Disease in MRL/lpr Mice

Cited by 7 publications

References 14 publications

Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Effect of MX-68 on airway inflammation and hyperresponsiveness in mice and guinea-pigs

Binding and transport of methotrexate and its derivative, MX-68, across the brush-border membrane in rat kidney

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A Novel Antifolate, MX-68, Inhibits the Development of Autoimmune Disease in MRL/lpr Mice

Cited by 7 publications

References 14 publications

Adenosine A2A or A3 receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Adenosine A2A or A3 receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Effect of MX-68 on airway inflammation and hyperresponsiveness in mice and guinea-pigs

Binding and transport of methotrexate and its derivative, MX-68, across the brush-border membrane in rat kidney

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Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68

Adenosine A_2A or A₃ receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68