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 P7C3-type aminopropylcarbazole compounds, which have significant neuroprotective effects on newly formed neural progenitor cells in the adult hippocampus and mature neurons in other regions of the central nervous system. This study is the first to examine whether the highly active compound P7C3-A20 has neuroprotective effects, promotes hippocampal neurogenesis, and improves functional outcomes after experimental TBI. Quantitative immunohistochemistry and behavioral assessments were performed on Sprague-Dawley rats that underwent moderate fluid-induced brain injury to observe post-traumatic changes. P7C3-A20 (10 mg/kg) or its solvent was administered intraperitoneally 30 minutes post-surgery, twice daily for 7 days. Following administration of P7C3-A20, the total volume of brain contusion was significantly reduced, vulnerable anti-neuronal nucleus (NeuN)-positive cortical neurons surrounding the contusion were protected, and sensorimotor function improved one week post-traumatically. P7C3-A20 treatment also significantly increased the number of bromideoxyuridine (BrdU) and dicortin (DCX) positive cells in the ipsilateral subgranular region of the dentate gyrus, a result that also occurred 1 week after traumatic brain injury (TBI). 5 weeks after TBI, compared with TBI control animals, the number of BrdU/NeuN double-labeled neurons in the P7C3-A20 treatment group was significantly increased, and cognitive function was improved in the Morris water maze test. These results suggest that P7C3-A20 has a neuroprotective effect and can promote endogenous repair mechanisms after TBI. We propose that the chemical framework represented by P7C3-A20 lays the foundation for optimizing and developing novel drugs that can protect patients from the early and chronic sequelae of traumatic brain injury (TBI). [2] Enhancing hippocampal neurogenesis is a potential new strategy for treating depression. Here, we validated this possibility by comparing hippocampal neurogenesis in ghrelin-releasing peptide receptor (Ghsr) knockout mice and wild-type littermates, and by measuring the antidepressant efficacy of P7C3-like neuroprotective compounds. Compared to wild-type littermates exposed to chronic social frustration disorder (CSDS), Ghsr knockout mice exhibited more severe depressive-like behaviors; however, 60% calorie restriction in Ghsr knockout mice failed to induce antidepressant-like behaviors. Chronic stress disorder (CSDS) significantly reduced cell proliferation and survival in the subgranular region of the ventral dentate gyrus (DG) in Ghsr knockout mice compared to wild-type littermates. Furthermore, calorie restriction increased apoptosis in the subgranular region of the DG in Ghsr knockout mice, while producing the opposite effect in wild-type littermates. Systemic administration of P7C3 during CSDS improved the survival of proliferating DG cells, which eventually developed into mature (NeuN+) neurons. It is noteworthy that P7C3 exhibited significant antidepressant-like effects in Ghsr gene knockout mice exposed to CSDS or calorie restriction, while the more active analog P7C3-A20 also showed antidepressant-like effects in wild-type littermates. Local ablation of hippocampal stem cells with radiation eliminated this antidepressant effect, further confirming that the mechanism of action of P7C3 antidepressants lies in their neuroprotective properties and the resulting promotion of hippocampal neurogenesis. In addition, the neurogenesis-promoting effect of P7C3-A20 is superior to that of many currently marketed antidepressants. In summary, our data confirm that abnormal hippocampal neurogenesis is an important factor in the etiology of depression and suggest that P7C3 compounds with neuroprotective effects may provide a new strategy for treating 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.)