Neuroprotective Properties of Mildronate, a Small Molecule, in a Rat Model of Parkinson’s Disease

Previously, we have found that mildronate [3-(2,2,2-trimethylhydrazinium) propionate dihydrate], a small molecule with charged nitrogen and oxygen atoms, protects mitochondrial metabolism that is altered by inhibitors of complex I and has neuroprotective effects in an azidothymidine-neurotoxicity mouse model. In the present study, we investigated the effects of mildronate in a rat model of Parkinson’s disease (PD) that was generated via a unilateral intrastriatal injection of the neurotoxin 6-hydroxydopamine (6-OHDA). We assessed the expression of cell biomarkers that are involved in signaling cascades and provide neural and glial integration: the neuronal marker TH (tyrosine hydroxylase); ubiquitin (a regulatory peptide involved in the ubiquitin-proteasome degradation system); Notch-3 (a marker of progenitor cells); IBA-1 (a marker of microglial cells); glial fibrillary acidic protein, GFAP (a marker of astrocytes); and inducible nitric oxide synthase, iNOS (a marker of inflammation). The data show that in the 6-OHDA-lesioned striatum, mildronate completely prevented the loss of TH, stimulated Notch-3 expression and decreased the expression of ubiquitin, GFAP and iNOS. These results provide evidence for the ability of mildronate to control the expression of an array of cellular proteins and, thus, impart multi-faceted homeostatic mechanisms in neurons and glial cells in a rat model of PD. We suggest that the use of mildronate provides a protective effect during the early stages of PD that can delay or halt the progression of this neurodegenerative disease

Матер. III съезда фармакологов России, 23-27 сентября 2007 г. (Тез 533)

Pupure J., Fernandes M.A.S., Oliveira C.R., Moreno A.J.M., Santos M.S., Kalvinsh I., Isajevs S., Klusa V. Mitochndria-regulating effects of mildronate in azidothymidine-induced toxicity Proceedings of the III Congress of Pharmacology Russia, Saint-Petersburg, 23–27 September 2007. Abstr 533. Psychopharmacol Biol Narcol 2007 Sept; 7 Suppl (Pt 2, M–Ja): 2-1911–2-1911 [in English]AZT (azidothymidine) is the first antiMHIV drug used in AIDS treatment, and it is still most commonly used in drug combinations. AZT inhibits polymerase g which is responsible for HIVM1 mitochondrial DNA replication, changes the structure of mitochondria by its incorporation into mitochondria [Peters et al., 1993]. Therefore the use of AZT is limited due to its severe side effects — myopathies, especially cardiomyopathies, lipodystrophy, hepatic steatosis, lactic acidosis and others [Badley et al., 2003]. We have suggested that AZTMinduced toxicity may be prevented by drugs capable of regulating mitochondrial processes. Hence, we have used mildronate, a cardioprotective drug of azaMbutyrobetaine class, which was shown to protect rat heart mitochondria from free fatty acid detergentMlike action by inhibiting carnitine palmitoyl transferase 1 [Dambrova et al., 2002]. Besides, mildronate protects the energy metabolism against the H2O2Minduced derangement in isolated rat heart (Akahira et al., 1997) and improves cardiac sarcoplasmic reticulum Ca2+ uptake activity [Hayashi et al., 2000]. In the present study, AZT and mildronate were administered in male mice intraperitoneally, per se and in combinations during a twoMweek period at doses previously found as the most active ones: AZT 50 mg/kg, and mildronate 50, 100 and 200 mg/kg. After termination of drug administration, mice were sacrificed and their heart and brain tissue were removed. Immunohistochemical assessment of nuclear factor kappa B (NFMkBp65) in the heart, and morphological assessment of heart and brain tissue was performed. The obtained results showed that AZT treatment significantly increased NFMkBp65 expression, indicating manifestation of oxidative stress leading to mitochondrial damage. AZT caused also morphological — inflammatory and degenerative — alterations in mice heart tissue. In brain tissue AZT caused degeneration of neurons in the molecular layer and external granular layer. Mildronate in combination with AZT reduce these AZTMinduced pathologic actions both in heart and brain tissue, demonstrating its powerful protective action. One may suggest that mildronate’s protective activity is provided via targeting mitochondrial processes. That indicates usefulness of mildronate’s combination with antiMHIV and other mitochondriaMcompromized drugs.  Pupure J., Fernandes M.A.S., Oliveira C.R., Moreno A.J.M., Santos M.S., Kalvinsh I., Isajevs S., Klusa V. . Матер. III съезда фармакологов России. Психофармакол биол наркол 2007 Спец Вып; 7 (Ч 2, М–Я): 2-1911–2-1911 (Тез 533

Матер. III съезда фармакологов России, 23-27 сентября 2007 г. (Тез 533)

Pupure J., Fernandes M.A.S., Oliveira C.R., Moreno A.J.M., Santos M.S., Kalvinsh I., Isajevs S., Klusa V. Mitochndria-regulating effects of mildronate in azidothymidine-induced toxicity Proceedings of the III Congress of Pharmacology Russia, Saint-Petersburg, 23–27 September 2007. Abstr 533. Psychopharmacol Biol Narcol 2007 Sept; 7 Suppl (Pt 2, M–Ja): 2-1911–2-1911 [in English]AZT (azidothymidine) is the first antiMHIV drug used in AIDS treatment, and it is still most commonly used in drug combinations. AZT inhibits polymerase g which is responsible for HIVM1 mitochondrial DNA replication, changes the structure of mitochondria by its incorporation into mitochondria [Peters et al., 1993]. Therefore the use of AZT is limited due to its severe side effects — myopathies, especially cardiomyopathies, lipodystrophy, hepatic steatosis, lactic acidosis and others [Badley et al., 2003]. We have suggested that AZTMinduced toxicity may be prevented by drugs capable of regulating mitochondrial processes. Hence, we have used mildronate, a cardioprotective drug of azaMbutyrobetaine class, which was shown to protect rat heart mitochondria from free fatty acid detergentMlike action by inhibiting carnitine palmitoyl transferase 1 [Dambrova et al., 2002]. Besides, mildronate protects the energy metabolism against the H2O2Minduced derangement in isolated rat heart (Akahira et al., 1997) and improves cardiac sarcoplasmic reticulum Ca2+ uptake activity [Hayashi et al., 2000]. In the present study, AZT and mildronate were administered in male mice intraperitoneally, per se and in combinations during a twoMweek period at doses previously found as the most active ones: AZT 50 mg/kg, and mildronate 50, 100 and 200 mg/kg. After termination of drug administration, mice were sacrificed and their heart and brain tissue were removed. Immunohistochemical assessment of nuclear factor kappa B (NFMkBp65) in the heart, and morphological assessment of heart and brain tissue was performed. The obtained results showed that AZT treatment significantly increased NFMkBp65 expression, indicating manifestation of oxidative stress leading to mitochondrial damage. AZT caused also morphological — inflammatory and degenerative — alterations in mice heart tissue. In brain tissue AZT caused degeneration of neurons in the molecular layer and external granular layer. Mildronate in combination with AZT reduce these AZTMinduced pathologic actions both in heart and brain tissue, demonstrating its powerful protective action. One may suggest that mildronate’s protective activity is provided via targeting mitochondrial processes. That indicates usefulness of mildronate’s combination with antiMHIV and other mitochondriaMcompromized drugs.  Pupure J., Fernandes M.A.S., Oliveira C.R., Moreno A.J.M., Santos M.S., Kalvinsh I., Isajevs S., Klusa V. . Матер. III съезда фармакологов России. Психофармакол биол наркол 2007 Спец Вып; 7 (Ч 2, М–Я): 2-1911–2-1911 (Тез 533

Ethyl 4-(5-bromo-2-hydroxyphenyl)-2,7,7-trimethyl-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate

In the title compound, C21H24BrNO4, the dihedral angle between the heterocyclic ring and the pendant aromatic ring is 80.20 (13)°. The hexahydroquinone [i.e. the one with the C=O group] ring adopts a sofa conformation. An intramolecular O—H...O hydrogen bond generates an S(6) ring motif. The ethyl group is disordered over two sets of sites with a refined site occupancy ratio of 0.633 (10):0.366 (10). In the crystal, molecules are linked by N—H...O interactions, forming chains parallel to [101]. There are no significant C—H...π or π–π interactions in the crystal structure

Characterization of the transporterB0AT3 (Slc6a17) in the rodent central nervous system

Background: The vesicular B(0)AT3 transporter (SLC6A17), one of the members of the SLC6 family, is a transporter for neutral amino acids and is exclusively expressed in brain. Here we provide a comprehensive expression profile of B(0)AT3 in mouse brain using in situ hybridization and immunohistochemistry. Results: We confirmed previous expression data from rat brain and used a novel custom made antibody to obtain detailed co-labelling with several cell type specific markers. B(0)AT3 was highly expressed in both inhibitory and excitatory neurons. The B(0)AT3 expression was highly overlapping with those of vesicular glutamate transporter 2 (VGLUT2) and vesicular glutamate transporter 1 (VGLUT1). We also show here that Slc6a17mRNA is up-regulated in animals subjected to short term food deprivation as well as animals treated with the serotonin reuptake inhibitor fluoxetine and the dopamine/noradrenaline reuptake inhibitor bupropion. Conclusions: This suggests that the B(0)AT3 transporter have a role in regulation of monoaminergic as well as glutamatergic synapses