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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

The MGV-1 derivatives 2-Cl-MGV-1 and 2-Cl-MGV-2 increase the lifespan of R6-2 mice, which are a transgenic animal model for Huntington disease. (a) The quinazoline-based, tricyclic compound 2-Cl-MGV-1 (which includes the Cl halogen substituent on the third rotatable carbocycle, that is, a halogenated MGV-1) increases median lifespan of R6-2 mice (n=5), compared with the vehicle DMSO-treated R6-2 mice (n=5). In more detail, 50% of untreated mice died before their 11th week, whereas 50% of the 2-Cl-MGV-1-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-1-treated mice). Applying Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-1-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 40% survival of the control mice (week 12 of age), the difference between the 2-Cl-MGV-1-treated R6-2 mice and the vehicle-injected R6-2 mice is significant: P<0.05 for week 12. Linear regression applied to weeks 8–13 shows that the intercepts are not equal (F=11.7, P<0.01), that is, 2-Cl-MGV-1-treated R6-2 mice start dying significantly later than vehicle-injected R6-2 mice. The slopes are very equal (F=3.23, NS), that is, at any given point in time fewer treated animals have died than untreated. (b) The quinazoline-based, tricyclic compound 2-Cl-MGV-2 (which in addition to the halogenation of 2-Cl-MGV-1, includes asymmetrical side chains, methyl and ethyl) increases median lifespan of the R6-2 mice (n=12), compared with the vehicle-treated R6-2 mice (n=10, control). In more detail, 50% of vehicle-treated R6-2 mice died before their 10th week, whereas 50% of the 2-Cl-MGV-2-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-2-treated mice). Applying ANOVA and Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-2-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 50% survival of the 2-Cl-MGV-2-treated R6-2 mice (week 12 of age) and in the week after that, the differences between the 2-Cl-MGV-2-treated R6-2 mice and the vehicle-injected R6-2 mice were significant P<0.05 for each of these two weeks. To determine whether the death rate of the vehicle-injected R6-2 mice is steeper than the death rate of 2-Cl-MGV-2-treated R6-2 mice we applied linear regression. Looking over the whole survival periods of both groups (F=12.5), and also over the restricted period from the week of diagnosis (week 8, after which the first animals have died) till all of the vehicle-treated R6-2 mice have died (week 13) (F=20.8), in both instances a significant difference between slopes is seen, P<0.01. The intercepts are not significantly different, meaning that the animals of the vehicle-treated and -untreated animals start dying from the same age. Thus, it appears that 2-Cl-MGV-1 is the superior agent for treatment of this animal model for Huntington disease.
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fig7: The MGV-1 derivatives 2-Cl-MGV-1 and 2-Cl-MGV-2 increase the lifespan of R6-2 mice, which are a transgenic animal model for Huntington disease. (a) The quinazoline-based, tricyclic compound 2-Cl-MGV-1 (which includes the Cl halogen substituent on the third rotatable carbocycle, that is, a halogenated MGV-1) increases median lifespan of R6-2 mice (n=5), compared with the vehicle DMSO-treated R6-2 mice (n=5). In more detail, 50% of untreated mice died before their 11th week, whereas 50% of the 2-Cl-MGV-1-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-1-treated mice). Applying Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-1-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 40% survival of the control mice (week 12 of age), the difference between the 2-Cl-MGV-1-treated R6-2 mice and the vehicle-injected R6-2 mice is significant: P<0.05 for week 12. Linear regression applied to weeks 8–13 shows that the intercepts are not equal (F=11.7, P<0.01), that is, 2-Cl-MGV-1-treated R6-2 mice start dying significantly later than vehicle-injected R6-2 mice. The slopes are very equal (F=3.23, NS), that is, at any given point in time fewer treated animals have died than untreated. (b) The quinazoline-based, tricyclic compound 2-Cl-MGV-2 (which in addition to the halogenation of 2-Cl-MGV-1, includes asymmetrical side chains, methyl and ethyl) increases median lifespan of the R6-2 mice (n=12), compared with the vehicle-treated R6-2 mice (n=10, control). In more detail, 50% of vehicle-treated R6-2 mice died before their 10th week, whereas 50% of the 2-Cl-MGV-2-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-2-treated mice). Applying ANOVA and Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-2-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 50% survival of the 2-Cl-MGV-2-treated R6-2 mice (week 12 of age) and in the week after that, the differences between the 2-Cl-MGV-2-treated R6-2 mice and the vehicle-injected R6-2 mice were significant P<0.05 for each of these two weeks. To determine whether the death rate of the vehicle-injected R6-2 mice is steeper than the death rate of 2-Cl-MGV-2-treated R6-2 mice we applied linear regression. Looking over the whole survival periods of both groups (F=12.5), and also over the restricted period from the week of diagnosis (week 8, after which the first animals have died) till all of the vehicle-treated R6-2 mice have died (week 13) (F=20.8), in both instances a significant difference between slopes is seen, P<0.01. The intercepts are not significantly different, meaning that the animals of the vehicle-treated and -untreated animals start dying from the same age. Thus, it appears that 2-Cl-MGV-1 is the superior agent for treatment of this animal model for Huntington disease.

Mentions: Surprisingly, MGV-1 did not appear to have an effect in the transgenic mouse model (R6-2 mice) for the hereditary human disease of Huntington, regarding behavior and lifespan.20 Also the classical TSPO ligand PK 11195 did not show such effects. However, derivatives from MGV-1 considerably and consistently extended average lifespan of these mice by 20%. In particular, in Figures 7a and b the effects of 2-Cl-MGV-1 and 2-Cl-MGV-2 on R6-2 mice are shown. MGV-2 had effects reminiscent of those of 2-Cl-MGV-1 and 2-Cl-MGV-2, but less consistent (data not shown). Interestingly, behavioral data suggested a reduction of uncontrollable tremor activity of R6-2 mice treated with 2-Cl-MGV-1 and 2-Cl-MGV-2, indicative of positive effects on motor control (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
The MGV-1 derivatives 2-Cl-MGV-1 and 2-Cl-MGV-2 increase the lifespan of R6-2 mice, which are a transgenic animal model for Huntington disease. (a) The quinazoline-based, tricyclic compound 2-Cl-MGV-1 (which includes the Cl halogen substituent on the third rotatable carbocycle, that is, a halogenated MGV-1) increases median lifespan of R6-2 mice (n=5), compared with the vehicle DMSO-treated R6-2 mice (n=5). In more detail, 50% of untreated mice died before their 11th week, whereas 50% of the 2-Cl-MGV-1-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-1-treated mice). Applying Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-1-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 40% survival of the control mice (week 12 of age), the difference between the 2-Cl-MGV-1-treated R6-2 mice and the vehicle-injected R6-2 mice is significant: P<0.05 for week 12. Linear regression applied to weeks 8–13 shows that the intercepts are not equal (F=11.7, P<0.01), that is, 2-Cl-MGV-1-treated R6-2 mice start dying significantly later than vehicle-injected R6-2 mice. The slopes are very equal (F=3.23, NS), that is, at any given point in time fewer treated animals have died than untreated. (b) The quinazoline-based, tricyclic compound 2-Cl-MGV-2 (which in addition to the halogenation of 2-Cl-MGV-1, includes asymmetrical side chains, methyl and ethyl) increases median lifespan of the R6-2 mice (n=12), compared with the vehicle-treated R6-2 mice (n=10, control). In more detail, 50% of vehicle-treated R6-2 mice died before their 10th week, whereas 50% of the 2-Cl-MGV-2-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-2-treated mice). Applying ANOVA and Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-2-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 50% survival of the 2-Cl-MGV-2-treated R6-2 mice (week 12 of age) and in the week after that, the differences between the 2-Cl-MGV-2-treated R6-2 mice and the vehicle-injected R6-2 mice were significant P<0.05 for each of these two weeks. To determine whether the death rate of the vehicle-injected R6-2 mice is steeper than the death rate of 2-Cl-MGV-2-treated R6-2 mice we applied linear regression. Looking over the whole survival periods of both groups (F=12.5), and also over the restricted period from the week of diagnosis (week 8, after which the first animals have died) till all of the vehicle-treated R6-2 mice have died (week 13) (F=20.8), in both instances a significant difference between slopes is seen, P<0.01. The intercepts are not significantly different, meaning that the animals of the vehicle-treated and -untreated animals start dying from the same age. Thus, it appears that 2-Cl-MGV-1 is the superior agent for treatment of this animal model for Huntington disease.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig7: The MGV-1 derivatives 2-Cl-MGV-1 and 2-Cl-MGV-2 increase the lifespan of R6-2 mice, which are a transgenic animal model for Huntington disease. (a) The quinazoline-based, tricyclic compound 2-Cl-MGV-1 (which includes the Cl halogen substituent on the third rotatable carbocycle, that is, a halogenated MGV-1) increases median lifespan of R6-2 mice (n=5), compared with the vehicle DMSO-treated R6-2 mice (n=5). In more detail, 50% of untreated mice died before their 11th week, whereas 50% of the 2-Cl-MGV-1-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-1-treated mice). Applying Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-1-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 40% survival of the control mice (week 12 of age), the difference between the 2-Cl-MGV-1-treated R6-2 mice and the vehicle-injected R6-2 mice is significant: P<0.05 for week 12. Linear regression applied to weeks 8–13 shows that the intercepts are not equal (F=11.7, P<0.01), that is, 2-Cl-MGV-1-treated R6-2 mice start dying significantly later than vehicle-injected R6-2 mice. The slopes are very equal (F=3.23, NS), that is, at any given point in time fewer treated animals have died than untreated. (b) The quinazoline-based, tricyclic compound 2-Cl-MGV-2 (which in addition to the halogenation of 2-Cl-MGV-1, includes asymmetrical side chains, methyl and ethyl) increases median lifespan of the R6-2 mice (n=12), compared with the vehicle-treated R6-2 mice (n=10, control). In more detail, 50% of vehicle-treated R6-2 mice died before their 10th week, whereas 50% of the 2-Cl-MGV-2-treated R6-2 mice were still alive until the 12th week. The y axis presents the percentage of surviving animals per week. The x axis presents the number of weeks from birth. The 50% survival cut off is marked with a horizontal arrow. The week where the 50% survival cut off is reached is marked with a vertical arrow (gray for control; black for 2-Cl-MGV-2-treated mice). Applying ANOVA and Wilcoxon matched-pairs signed rank test regarding the number of surviving animals indicates a significant difference between the 2-Cl-MGV-2-treated R6-2 mice and the DMSO (vehicle)-injected R6-2 mice: P<0.01. Applying Mann–Whitney to each week of treatment shows that at the week of 50% survival of the 2-Cl-MGV-2-treated R6-2 mice (week 12 of age) and in the week after that, the differences between the 2-Cl-MGV-2-treated R6-2 mice and the vehicle-injected R6-2 mice were significant P<0.05 for each of these two weeks. To determine whether the death rate of the vehicle-injected R6-2 mice is steeper than the death rate of 2-Cl-MGV-2-treated R6-2 mice we applied linear regression. Looking over the whole survival periods of both groups (F=12.5), and also over the restricted period from the week of diagnosis (week 8, after which the first animals have died) till all of the vehicle-treated R6-2 mice have died (week 13) (F=20.8), in both instances a significant difference between slopes is seen, P<0.01. The intercepts are not significantly different, meaning that the animals of the vehicle-treated and -untreated animals start dying from the same age. Thus, it appears that 2-Cl-MGV-1 is the superior agent for treatment of this animal model for Huntington disease.
Mentions: Surprisingly, MGV-1 did not appear to have an effect in the transgenic mouse model (R6-2 mice) for the hereditary human disease of Huntington, regarding behavior and lifespan.20 Also the classical TSPO ligand PK 11195 did not show such effects. However, derivatives from MGV-1 considerably and consistently extended average lifespan of these mice by 20%. In particular, in Figures 7a and b the effects of 2-Cl-MGV-1 and 2-Cl-MGV-2 on R6-2 mice are shown. MGV-2 had effects reminiscent of those of 2-Cl-MGV-1 and 2-Cl-MGV-2, but less consistent (data not shown). Interestingly, behavioral data suggested a reduction of uncontrollable tremor activity of R6-2 mice treated with 2-Cl-MGV-1 and 2-Cl-MGV-2, indicative of positive effects on motor control (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&thinsp;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 &lsquo;neuronal nuclei&rsquo;, 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 &gt;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