Neuroprotective agent

In PC12 cells, P7C3-A20 (10-100 μM; 8 hours) treatment lessens the cytotoxicity generated by oxygen-reduction enhanced (OGD) [1].

In the HI paradigm, P7C3-A20 (5–10 mg/kg; intraperitoneal; daily; 7 days; Sprague-Dawley rats) decreases the number of infarcts, reverses the loss of cells in the retina and hippocampus, and enhances motor function. However, P7C3–A20 cannot stop HI-induced neuronal damage by turning on PI3K/AKT/GSK3β signaling [1].

Hypoxic-ischemic encephalopathy (HIE) in neonates can lead to severe long-term disabilities including cerebral palsy and brain injury. The small molecule P7C3-A20 has been shown to exert neuroprotective effects in various disorders such as ischemic stroke and neurodegenerative diseases. However, it is unclear whether P7C3-A20 has therapeutic potential for the treatment of HIE, and the relationship between P7C3-A20 and neuronal apoptosis is unknown. To address these questions, the present study investigated whether P7C3-A20 reduces HI injury in vitro using a PC12 cell oxygen-glucose deprivation (OGD) model and in vivo in postnatal day 7 and 14 rats subjected to HI, along with the underlying mechanisms. We found that treatment with P7C3-A20 (40-100 µM) alleviated OGD-induced apoptosis in PC12 cells. In HI model rats, treatment with 5 or 10 mg/kg P7C3-A20 reduced infarct volume; reversed cell loss in the cortex and hippocampus and improved motor function without causing neurotoxicity. The neuroprotective effects were abrogated by treatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. These results demonstrate that P7C3-A20 exerts neuroprotection by activating PI3K/protein kinase B/glycogen synthase kinase 3β signaling and can potentially be used to prevent brain injury in neonates following HIE[1].

Cell Viability Assay[1]
Cell Types: PC12 cells
Tested Concentrations: 10 μM, 20 μM, 40 μM, 60 μM, 80 μM, 100 μM
Incubation Duration: 8 hrs (hours)) treatment can attenuate OGD-induced PC12 cell sterility[1].
Experimental Results: Mitigated oxygen glucose deprivation (OGD)-induced cytotoxicity in PC12 cells.

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Apoptosis analysis [1]
Cell Types: PC12 Cell
Tested Concentrations: 40 μM, 60 μM, 80 μM, 100 μM
Incubation Duration: 8 hrs (hours)
Experimental Results: Reduce oxygen glucose deprivation (OGD)-induced PC12 cell apoptosis.

Animal/Disease Models: SD (SD (Sprague-Dawley)) rat (200-250 g) induced hypoxic-ischemic (HI) injury [1]
Doses: 5 mg/kg, 10 mg/kg
Route of Administration: intraperitoneal (ip) injection; daily; continued for 7 Day
Experimental Results: Reduction in infarct volume; reversal of cell loss in cortex and hippocampus and improvement in motor function without causing neurotoxicity.

[1]. The Small Molecule P7C3-A20 Exerts Neuroprotective Effects in a Hypoxic-Ischemic Encephalopathy Model via Activation of PI3K/AKT/GSK3β Signaling. Neuroscience. 2020 Jun 3;S0306-4522(20)30353-5.

[2]. Neuroprotective efficacy of a proneurogenic compound after traumatic brain injury. J Neurotrauma. 2014 Mar 1;31(5):476-86.

[3]. The P7C3 class of neuroprotective compounds exerts antidepressant efficacy in mice by increasinghippocampal neurogenesis. Mol Psychiatry. 2015 Apr;20(4):500-8.

Traumatic brain injury (TBI) is characterized by histopathological damage and long-term sensorimotor and cognitive dysfunction. Recent studies have reported the discovery of the P7C3 class of aminopropyl carbazole agents with potent neuroprotective properties for both newborn neural precursor cells in the adult hippocampus and mature neurons in other regions of the central nervous system. This study tested, for the first time, whether the highly active P7C3-A20 compound would be neuroprotective, promote hippocampal neurogenesis, and improve functional outcomes after experimental TBI. Sprague-Dawley rats subjected to moderate fluid percussion brain injury were evaluated for quantitative immunohistochemical and behavioral changes after trauma. P7C3-A20 (10 mg/kg) or vehicle was initiated intraperitoneally 30 min postsurgery and twice per day every day thereafter for 7 days. Administration of P7C3-A20 significantly reduced overall contusion volume, preserved vulnerable anti-neuronal nuclei (NeuN)-positive pericontusional cortical neurons, and improved sensorimotor function 1 week after trauma. P7C3-A20 treatment also significantly increased both bromodeoxyuridine (BrdU)- and doublecortin (DCX)-positive cells within the subgranular zone of the ipsilateral dentate gyrus 1 week after TBI. Five weeks after TBI, animals treated with P7C3-A20 showed significantly increased BrdU/NeuN double-labeled neurons and improved cognitive function in the Morris water maze, compared to TBI-control animals. These results suggest that P7C3-A20 is neuroprotective and promotes endogenous reparative strategies after TBI. We propose that the chemical scaffold represented by P7C3-A20 provides a basis for optimizing and advancing new pharmacological agents for protecting patients against the early and chronic consequences of TBI.[2]

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Augmenting hippocampal neurogenesis represents a potential new strategy for treating depression. Here we test this possibility by comparing hippocampal neurogenesis in depression-prone ghrelin receptor (Ghsr)-null mice to that in wild-type littermates and by determining the antidepressant efficacy of the P7C3 class of neuroprotective compounds. Exposure of Ghsr-null mice to chronic social defeat stress (CSDS) elicits more severe depressive-like behavior than in CSDS-exposed wild-type littermates, and exposure of Ghsr-null mice to 60% caloric restriction fails to elicit antidepressant-like behavior. CSDS resulted in more severely reduced cell proliferation and survival in the ventral dentate gyrus (DG) subgranular zone of Ghsr-null mice than in that of wild-type littermates. Also, caloric restriction increased apoptosis of DG subgranular zone cells in Ghsr-null mice, although it had the opposite effect in wild-type littermates. Systemic treatment with P7C3 during CSDS increased survival of proliferating DG cells, which ultimately developed into mature (NeuN+) neurons. Notably, P7C3 exerted a potent antidepressant-like effect in Ghsr-null mice exposed to either CSDS or caloric restriction, while the more highly active analog P7C3-A20 also exerted an antidepressant-like effect in wild-type littermates. Focal ablation of hippocampal stem cells with radiation eliminated this antidepressant effect, further attributing the P7C3 class antidepressant effect to its neuroprotective properties and resultant augmentation of hippocampal neurogenesis. Finally, P7C3-A20 demonstrated greater proneurogenic efficacy than a wide spectrum of currently marketed antidepressant drugs. Taken together, our data confirm the role of aberrant hippocampal neurogenesis in the etiology of depression and suggest that the neuroprotective P7C3-compounds represent a novel strategy for treating patients with this disease.[3]

C22H19BR2FN2O

506.20546746254

503.984

C, 52.20; H, 3.78; Br, 31.57; F, 3.75; N, 5.53; O, 3.16

1235481-90-9

P7C3;301353-96-8

46853447

Typically exists as White to yellow solids at room temperature

1.6±0.1 g/cm3

641.3±55.0 °C at 760 mmHg

341.7±31.5 °C

0.0±1.9 mmHg at 25°C

1.647

6.96

1

3

6

28

487

0

FC(CNC1=CC(OC)=CC=C1)CN2C3=CC=C(Br)C=C3C4=CC(Br)=CC=C24

XNLTWMQBJFWQOU-UHFFFAOYSA-N

InChI=1S/C22H19Br2FN2O/c1-28-18-4-2-3-17(11-18)26-12-16(25)13-27-21-7-5-14(23)9-19(21)20-10-15(24)6-8-22(20)27/h2-11,16,26H,12-13H2,1H3

N-[3-(3,6-dibromocarbazol-9-yl)-2-fluoropropyl]-3-methoxyaniline

P7C3-A20; 1235481-90-9; N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-fluoropropyl)-3-methoxyaniline; N-[3-(3,6-dibromocarbazol-9-yl)-2-fluoropropyl]-3-methoxyaniline; CHEMBL2442625; N-[3-(3,6-dibromo-9H-carbazol-9-yl)-2-fluoropropyl]-3-methoxyaniline; 1235481-90-9 (free base); 9H-Carbazole-9-propanamine, 3,6-dibromo-beta-fluoro-N-(3-methoxyphenyl)-; P7C3A20; P7C3 A20

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 100 mg/mL (~197.55 mM)

Solubility in Formulation 1: ≥ 3.85 mg/mL (7.61 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 38.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (4.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)