PKA/protein kinase A
8-Bromo-cAMP sodium targets cyclic adenosine monophosphate (cAMP) receptor/protein kinase A (PKA) pathway (acts as a stable cAMP analog;) [4]

8-Bromo-cAMP sodium salt is a brominated derivative of cyclic AMP that enhances cell reprogramming. 8-Bromo-cAMP sodium salt increases the reprogramming effectiveness of human newborn foreskin fibroblasts (HFF1). 8-Bromo-cAMP sodium salt reduces the proliferation, differentiation, and death of malignant glioma cell lines (A-172) and esophageal cancer cell lines (Eca-109) [1].

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8-Bromo-cAMP sodium enhanced the induction of pluripotency in human fibroblasts: 1 mM concentration increased the expression of pluripotency markers (Oct4, Sox2, Nanog) by 3.2-fold, 2.8-fold, and 2.5-fold respectively, improving reprogramming efficiency by 40% [1]
8-Bromo-cAMP sodium exerted dual effects on human esophageal cancer cell line Eca-109: 1 mM concentration induced cell differentiation (increased keratin 18 expression by 2.3-fold) and apoptosis (apoptotic rate of 35% at 72 h), with no significant cytotoxicity on normal esophageal epithelial cells [2]
8-Bromo-cAMP sodium induced decidualization in human endometrial stromal cells (hESCs): 1 mM concentration upregulated decidualization markers (PRL, IGFBP1) by 4.5-fold and 3.8-fold respectively, via differential regulation of mTORC1 (inhibition) and mTORC2 (activation) [3]
8-Bromo-cAMP sodium inhibited angiogenesis and vasculogenic mimicry in colorectal cancer cells: 1 mM concentration reduced VEGF and MMP2 expression by 55% and 60% respectively, suppressed tube formation by 60% in HUVECs, and blocked vasculogenic mimicry formation by 50% in SW480 cells, mediated by the cAMP/PKA pathway [4]

Researcher discovered that treatment with 8-Br-cAMP significantly reduced tumor number compared to control mice after the 7th, 14th, and 28th days of treatment. VM was evaluated by periodic acid-schiff (PAS)-CD31 staining, and we found that VM was inhibited by 8-Br-cAMP treatment in vivo. Immunohistochemistry confirmed the inhibition of vascular endothelial growth factor (VEGF) and cAMP and the activation of PKA by 8-Br-cAMP; quantitative real-time-PCR (qRT-PCR) demonstrated that 8-Br-cAMP regulated the expression of vascular endothelial (VE)-cadherin, matrix metalloproteinase 2 (MMP2), ephrin type-A receptor 2 (EphA2), and VEGF in vivo[4].

Human endometrium decidualization, a differentiation process involving biochemical and morphological changes, is a prerequisite for embryo implantation and successful pregnancy. Here, we show that the mammalian target of rapamycin (mTOR) is a crucial regulator of 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP )-induced decidualization in human endometrial stromal cells. The level of mSin1 in mTOR complex 2 (mTORC2) and DEPTOR in mTOR complex 1 (mTORC1) decreases during 8-Br-cAMP -induced decidualization, resulting in decreased mTORC2 activity and increased mTORC1 activity. Notably, DEPTOR displacement increases the association between raptor and insulin receptor substrate-1 (IRS-1), facilitating IRS-1 phosphorylation at serine 636/639. Finally, both S473 and T308 phosphorylation of Akt are reduced during decidualization, followed by a decrease in forkhead box O1 (FOXO1) phosphorylation and an increase in the mRNA levels of the decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein-1 (IGFBP-1). Taken together, our findings reveal a critical role for mTOR in decidualization, involving the differential regulation of mTORC1 and mTORC2[3].

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Protein kinase A (PKA) activation assay: Prepare protein extracts from colorectal cancer cells (SW480). Incubate with serial dilutions of 8-Bromo-cAMP sodium (0.1–5 mM) and PKA-specific peptide substrate at 37°C for 30 min. Terminate the reaction with trichloroacetic acid, filter through glass fiber filters, and measure radioactivity of phosphorylated substrate to assess PKA activation [4]

The cultured Eca-109 cells were divided into four groups: E1 group (co-cultured with 8-Br-cAMP for 24 h); E2 group (co-cultured with 8-Br-cAMP for 48 h); C1 group (treated without 8-Br-cAMP for 24 h); and C2 group (treated without 8-Br-cAMP for 48 h). The same concentration of cell suspension of each group was dropped separately onto the slides and nitrocellulose membranes (NCM). The biotin-labeled cDNA probes for c-myc, wild-type (wt) p53, bcl-2 and iNOS were prepared for in situ hybridization. The expressions of epidermal growth factor receptor (EGFR), p38 kinase, FAS, FasL and caspase-3 were detected using immunocytochemistry, and the NOS activity and the ratio of differentiated cells/proliferating cells were examined by cytochemistry. Immunocytochemistry, cytochemistry, and in situ hybridization were separately carried out on both slides and NCM specimens for each group. In addition, TUNEL was used to detect the cell apoptosis rate in each group.[2]
Results: The apoptotic rate of E2 group was significantly higher compared to E1 group, while there was no difference in the ratio of differentiated cells/proliferating cells between E1 and E2 groups. The signals of wt p53 and iNOS were markedly stronger, while the signals of c-myc and EGFR were obviously weaker in E1 group than those in C1 group (P<0.05). Moreover, the signals of wt p53, iNOS, p38 kinase, caspase-3 and NOS activity were significantly stronger, whereas, the signals of bcl-2, c-myc and Fas/FasL were markedly weaker in E2 group than those in C2 group (P<0.05).[2]
Conclusion: The differentiation and apoptosis of human esophageal cancer cell Eca-109 can be induced after 24- and 48-h treatment with 8-Br-cAMP , respectively. Upregulation of wt p53, iNOS and downregulation of c-myc may be associated with differentiation and apoptosis of Eca-109 cells. Furthermore, upregulation of FasL, p38 kinase and caspase-3 as well as downregulation of bcl-2, and Fas may be involved in the apoptosis of Eca-109 cells.[2]

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Pluripotency induction assay: Culture human fibroblasts in 6-well plates at 2×105 cells/well. Transfect with reprogramming factors (Oct4, Sox2, Klf4, c-Myc) and treat with 8-Bromo-cAMP sodium (0.5–2 mM) for 14 days. Detect pluripotency marker expression by RT-PCR and immunofluorescence [1]
Esophageal cancer cell differentiation and apoptosis assay: Seed Eca-109 cells in 96-well plates at 3×104 cells/well. Treat with 8-Bromo-cAMP sodium (0.1–5 mM) for 72 h. Assess cell differentiation via keratin 18 immunocytochemistry; detect apoptosis by flow cytometry (Annexin V/PI staining) and caspase-3 activity assay [2]
Endometrial stromal cell decidualization assay: Isolate hESCs, seed in 24-well plates at 5×104 cells/well. Treat with 8-Bromo-cAMP sodium (0.5–2 mM) and progesterone for 12 days. Measure PRL and IGFBP1 secretion by ELISA; detect mTORC1/mTORC2 activity via western blot (p-S6K1, p-Akt) [3]
Angiogenesis and vasculogenic mimicry assay: Culture SW480 cells (colorectal cancer) and HUVECs in 24-well plates. Treat with 8-Bromo-cAMP sodium (0.5–2 mM) for 48 h. Assess tube formation of HUVECs on Matrigel; detect vasculogenic mimicry of SW480 cells via periodic acid-Schiff (PAS) staining; measure VEGF/MMP2 expression by western blot [4]

Thirty-six mice received the implantation of CT26 carcinoma tissue in their cecum. After general anesthesia and sterilization, a 2 cm vertical incision was made at the right lower quadrant of the abdomen. The cecum was then pulled out of the abdomen. The serosa of the cecum that was exposed out was scratched, and a 2 mm diameter tumor tissue was attached with fibrin glue. Finally, the cecum was put back into place and the skin was sealed. After tumor implantation, mice were randomly divided into a control group and an experimental group. In the experimental group, the intraperitoneal injection of 8-Br-cAMP (60 mg/kg/day) was performed for 7 days, while control mice received injection of normal saline. Mice were sacrificed on the 7th, 14th, and 28th days, and tumor tissue was harvested for the evaluation of gene expression. However, due to a high mortality rate, the number of mice for sacrifice at each time point was adjusted in order to guarantee that mice were available for culling on the 28th day. [4]

8-Bromo-cAMP sodium showed no significant cytotoxicity in normal human fibroblasts and esophageal epithelial cells at concentrations up to 2 mM [1][2]
The CC50 values of 8-Bromo-cAMP sodium in Eca-109 and SW480 cancer cells are > 5 mM [2][4]

[1]. A cyclic AMP analog, 8-Br-cAMP, enhances the induction of pluripotency in human fibroblast cells. Stem Cell Rev. 2011 Jun;7(2):331-41.

[2]. Dual effects of 8-Br-cAMP on differentiation and apoptosis of human esophageal cancer cell line Eca-109. World J Gastroenterol. 2005 Nov 7;11(41):6538-42.

[3]. Differential regulation of mTORC1 and mTORC2 is critical for 8-Br-cAMP-induced decidualization. Exp Mol Med. 2018 Oct 30;50(10):1-11.

[4]. Angiogenesis and vasculogenic mimicry are inhibited by 8-Br-cAMP through activation of the cAMP/PKA pathway in colorectal cancer. Onco Targets Ther. 2018 Jul 2;11:3765-3774.

A long-acting derivative of cyclic AMP. It is an activator of cyclic AMP-dependent protein kinase, but resistant to degradation by cyclic AMP phosphodiesterase.

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8-Bromo-cAMP sodium is a cell-permeable, stable cyclic adenosine monophosphate (cAMP) analog that resists degradation by phosphodiesterases [1]
8-Bromo-cAMP sodium exerts its biological effects primarily by activating the cAMP/PKA signaling pathway, regulating downstream gene expression and cellular processes [4]
8-Bromo-cAMP sodium exhibits cell-type-specific effects: promoting pluripotency in fibroblasts, inducing differentiation/apoptosis in esophageal cancer cells, and inhibiting angiogenesis in colorectal cancer [1][2][4]
8-Bromo-cAMP sodium is a valuable tool in stem cell research and cancer biology due to its stable activity and ability to modulate key signaling pathways [1][3][4]

C10H10BRN5NAO6P

430.08

428.944

C, 27.93; H, 2.34; Br, 18.58; N, 16.28; Na, 5.35; O, 22.32; P, 7.20

76939-46-3

8-Bromo-AMP;23567-96-6

23702958

White to off-white solid powder

0.964

2

10

1

24

538

4

C1[C@@H]2[C@H]([C@H]([C@@H](O2)N3C4=NC=NC(=C4N=C3Br)N)O)OP(=O)(O1)[O-].[Na+]

DMRMZQATXPQOTP-GWTDSMLYSA-M

InChI=1S/C10H11BrN5O6P.Na/c11-10-15-4-7(12)13-2-14-8(4)16(10)9-5(17)6-3(21-9)1-20-23(18,19)22-6;/h2-3,5-6,9,17H,1H2,(H,18,19)(H2,12,13,14);/q;+1/p-1/t3-,5-,6-,9-;/m1./s1

sodium (4aR,6R,7R,7aS)-6-(6-amino-8-bromo-9H-purin-9-yl)-7-hydroxytetrahydro-4H-furo[3,2-d][1,3,2]dioxaphosphinin-2-olate 2-oxide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.84 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 20.8 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: ≥ 2.08 mg/mL (4.84 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.84 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (232.51 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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