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Quinazoline-based tricyclic compounds that regulate programmed cell death, induce neuronal differentiation, and are curative in animal models for excitotoxicity and hereditary brain disease

View Article: PubMed Central - PubMed

ABSTRACT

Expanding on a quinazoline scaffold, we developed tricyclic compounds with biological activity. These compounds bind to the 18 kDa translocator protein (TSPO) and protect U118MG (glioblastoma cell line of glial origin) cells from glutamate-induced cell death. Fascinating, they can induce neuronal differentiation of PC12 cells (cell line of pheochromocytoma origin with neuronal characteristics) known to display neuronal characteristics, including outgrowth of neurites, tubulin expression, and NeuN (antigen known as ‘neuronal nuclei’, also known as Rbfox3) expression. As part of the neurodifferentiation process, they can amplify cell death induced by glutamate. Interestingly, the compound 2-phenylquinazolin-4-yl dimethylcarbamate (MGV-1) can induce expansive neurite sprouting on its own and also in synergy with nerve growth factor and with glutamate. Glycine is not required, indicating that N-methyl-D-aspartate receptors are not involved in this activity. These diverse effects on cells of glial origin and on cells with neuronal characteristics induced in culture by this one compound, MGV-1, as reported in this article, mimic the diverse events that take place during embryonic development of the brain (maintenance of glial integrity, differentiation of progenitor cells to mature neurons, and weeding out of non-differentiating progenitor cells). Such mechanisms are also important for protective, curative, and restorative processes that occur during and after brain injury and brain disease. Indeed, we found in a rat model of systemic kainic acid injection that MGV-1 can prevent seizures, counteract the process of ongoing brain damage, including edema, and restore behavior defects to normal patterns. Furthermore, in the R6-2 (transgenic mouse model for Huntington disease; Strain name: B6CBA-Tg(HDexon1)62Gpb/3J) transgenic mouse model for Huntington disease, derivatives of MGV-1 can increase lifespan by >20% and reduce incidence of abnormal movements. Also in vitro, these derivatives were more effective than MGV-1.

No MeSH data available.


Related in: MedlinePlus

Localization of tubulin 3β (a–e) and NeuN (f–j) in cell bodies and neurites of differentiated PC12 (strain #3). This figure shows that our different protocols not only result in extensive sprouting and outgrowth of neurites of PC12 cells in culture (as shown in Figure 3), but also labeling of these cells with the neuronal markers tubulin 3β (magenta in a–e) and NeuN (yellow in f–j). The cell nuclei are labeled with DAPI (cyan in a–j). (a) Tubulin 3β labeling can be detected first of all in the cell bodies of the undifferentiated vehicle control PC12 cells (control). Inducing differentiation with MGV-1 (b), MGV-1 plus glutamate (c), NGF (d), as well as MGV-1 plus NGF plus glutamate (e) enhanced tubulin 3β labeling not only of the cell body but also intensely of neurites. (f) NeuN expression is indicated with yellow fluorescent immunocytochemical labeling of the cell bodies, both in the nuclei and the cytoplasm of undifferentiated cells (control). Nuclei and cytoplasm both are typical locations for NeuN.91 NeuN labeling can also appear in the neurites of cells differentiated with MGV-1 (g), MGV-1 plus glutamate (h), NGF (i), as well as MGV-1 plus NGF plus glutamate (j). NeuN labeling can also appear in the neurites. In undifferentiated as well as differentiated cells doubly labeled for DAPI and NeuN, the cell nuclei can appear whitish, indicating the presence of NeuN in the cell nuclei. The same is true Tubulin for cells doubly labeled for DAPI and tubulin. The scale bars in are 100 μM.
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fig4: Localization of tubulin 3β (a–e) and NeuN (f–j) in cell bodies and neurites of differentiated PC12 (strain #3). This figure shows that our different protocols not only result in extensive sprouting and outgrowth of neurites of PC12 cells in culture (as shown in Figure 3), but also labeling of these cells with the neuronal markers tubulin 3β (magenta in a–e) and NeuN (yellow in f–j). The cell nuclei are labeled with DAPI (cyan in a–j). (a) Tubulin 3β labeling can be detected first of all in the cell bodies of the undifferentiated vehicle control PC12 cells (control). Inducing differentiation with MGV-1 (b), MGV-1 plus glutamate (c), NGF (d), as well as MGV-1 plus NGF plus glutamate (e) enhanced tubulin 3β labeling not only of the cell body but also intensely of neurites. (f) NeuN expression is indicated with yellow fluorescent immunocytochemical labeling of the cell bodies, both in the nuclei and the cytoplasm of undifferentiated cells (control). Nuclei and cytoplasm both are typical locations for NeuN.91 NeuN labeling can also appear in the neurites of cells differentiated with MGV-1 (g), MGV-1 plus glutamate (h), NGF (i), as well as MGV-1 plus NGF plus glutamate (j). NeuN labeling can also appear in the neurites. In undifferentiated as well as differentiated cells doubly labeled for DAPI and NeuN, the cell nuclei can appear whitish, indicating the presence of NeuN in the cell nuclei. The same is true Tubulin for cells doubly labeled for DAPI and tubulin. The scale bars in are 100 μM.

Mentions: We also applied our treatments to cells with the more standard looking PC12 cells, strains #2 and #3. We could differentiate these cells by NGF by itself as described previously by others (Figures 3d and 4). A new finding is that we could also differentiate these cells with glutamate by itself (Figures 3d and 4). Moreover, these cells could be differentiated by MGV-1 by itself as well. We also found that NGF, MGV-1, and glutamate can enhance each other's differentiating capabilities in these cells (Figure 3d). In applications with glutamate shown here, glycine is routinely applied. Noteworthy, however, omission of glycine does not modify the response of any of the cell strains (data not shown). Similarly, comparing the vehicle controls with and without alcohol (1% final concentration) also does not show a difference whatsoever (data not shown).


Quinazoline-based tricyclic compounds that regulate programmed cell death, induce neuronal differentiation, and are curative in animal models for excitotoxicity and hereditary brain disease
Localization of tubulin 3β (a–e) and NeuN (f–j) in cell bodies and neurites of differentiated PC12 (strain #3). This figure shows that our different protocols not only result in extensive sprouting and outgrowth of neurites of PC12 cells in culture (as shown in Figure 3), but also labeling of these cells with the neuronal markers tubulin 3β (magenta in a–e) and NeuN (yellow in f–j). The cell nuclei are labeled with DAPI (cyan in a–j). (a) Tubulin 3β labeling can be detected first of all in the cell bodies of the undifferentiated vehicle control PC12 cells (control). Inducing differentiation with MGV-1 (b), MGV-1 plus glutamate (c), NGF (d), as well as MGV-1 plus NGF plus glutamate (e) enhanced tubulin 3β labeling not only of the cell body but also intensely of neurites. (f) NeuN expression is indicated with yellow fluorescent immunocytochemical labeling of the cell bodies, both in the nuclei and the cytoplasm of undifferentiated cells (control). Nuclei and cytoplasm both are typical locations for NeuN.91 NeuN labeling can also appear in the neurites of cells differentiated with MGV-1 (g), MGV-1 plus glutamate (h), NGF (i), as well as MGV-1 plus NGF plus glutamate (j). NeuN labeling can also appear in the neurites. In undifferentiated as well as differentiated cells doubly labeled for DAPI and NeuN, the cell nuclei can appear whitish, indicating the presence of NeuN in the cell nuclei. The same is true Tubulin for cells doubly labeled for DAPI and tubulin. The scale bars in are 100 μM.
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Related In: Results  -  Collection

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fig4: Localization of tubulin 3β (a–e) and NeuN (f–j) in cell bodies and neurites of differentiated PC12 (strain #3). This figure shows that our different protocols not only result in extensive sprouting and outgrowth of neurites of PC12 cells in culture (as shown in Figure 3), but also labeling of these cells with the neuronal markers tubulin 3β (magenta in a–e) and NeuN (yellow in f–j). The cell nuclei are labeled with DAPI (cyan in a–j). (a) Tubulin 3β labeling can be detected first of all in the cell bodies of the undifferentiated vehicle control PC12 cells (control). Inducing differentiation with MGV-1 (b), MGV-1 plus glutamate (c), NGF (d), as well as MGV-1 plus NGF plus glutamate (e) enhanced tubulin 3β labeling not only of the cell body but also intensely of neurites. (f) NeuN expression is indicated with yellow fluorescent immunocytochemical labeling of the cell bodies, both in the nuclei and the cytoplasm of undifferentiated cells (control). Nuclei and cytoplasm both are typical locations for NeuN.91 NeuN labeling can also appear in the neurites of cells differentiated with MGV-1 (g), MGV-1 plus glutamate (h), NGF (i), as well as MGV-1 plus NGF plus glutamate (j). NeuN labeling can also appear in the neurites. In undifferentiated as well as differentiated cells doubly labeled for DAPI and NeuN, the cell nuclei can appear whitish, indicating the presence of NeuN in the cell nuclei. The same is true Tubulin for cells doubly labeled for DAPI and tubulin. The scale bars in are 100 μM.
Mentions: We also applied our treatments to cells with the more standard looking PC12 cells, strains #2 and #3. We could differentiate these cells by NGF by itself as described previously by others (Figures 3d and 4). A new finding is that we could also differentiate these cells with glutamate by itself (Figures 3d and 4). Moreover, these cells could be differentiated by MGV-1 by itself as well. We also found that NGF, MGV-1, and glutamate can enhance each other's differentiating capabilities in these cells (Figure 3d). In applications with glutamate shown here, glycine is routinely applied. Noteworthy, however, omission of glycine does not modify the response of any of the cell strains (data not shown). Similarly, comparing the vehicle controls with and without alcohol (1% final concentration) also does not show a difference whatsoever (data not shown).

View Article: PubMed Central - PubMed

ABSTRACT

Expanding on a quinazoline scaffold, we developed tricyclic compounds with biological activity. These compounds bind to the 18 kDa translocator protein (TSPO) and protect U118MG (glioblastoma cell line of glial origin) cells from glutamate-induced cell death. Fascinating, they can induce neuronal differentiation of PC12 cells (cell line of pheochromocytoma origin with neuronal characteristics) known to display neuronal characteristics, including outgrowth of neurites, tubulin expression, and NeuN (antigen known as ‘neuronal nuclei’, also known as Rbfox3) expression. As part of the neurodifferentiation process, they can amplify cell death induced by glutamate. Interestingly, the compound 2-phenylquinazolin-4-yl dimethylcarbamate (MGV-1) can induce expansive neurite sprouting on its own and also in synergy with nerve growth factor and with glutamate. Glycine is not required, indicating that N-methyl-D-aspartate receptors are not involved in this activity. These diverse effects on cells of glial origin and on cells with neuronal characteristics induced in culture by this one compound, MGV-1, as reported in this article, mimic the diverse events that take place during embryonic development of the brain (maintenance of glial integrity, differentiation of progenitor cells to mature neurons, and weeding out of non-differentiating progenitor cells). Such mechanisms are also important for protective, curative, and restorative processes that occur during and after brain injury and brain disease. Indeed, we found in a rat model of systemic kainic acid injection that MGV-1 can prevent seizures, counteract the process of ongoing brain damage, including edema, and restore behavior defects to normal patterns. Furthermore, in the R6-2 (transgenic mouse model for Huntington disease; Strain name: B6CBA-Tg(HDexon1)62Gpb/3J) transgenic mouse model for Huntington disease, derivatives of MGV-1 can increase lifespan by >20% and reduce incidence of abnormal movements. Also in vitro, these derivatives were more effective than MGV-1.

No MeSH data available.


Related in: MedlinePlus