Endogenous Metabolite; P2X7 receptor

Co-treating LPS (1 μg/mL) with ATP (5 mM) for one hour has a positive impact on HGFs' NLRP3 inflammasome activation [3]. IL-1β, KC, and MIP-2 are all required for the caspase-1 activation-dependent induction of BMDM mortality by ATP (2 mM; 0.5-24 hours) [4]. In vitro, ATP stimulates neutrophil chemotaxis [4].

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- Cytokine release modulation: Adenosine-5'-Triphosphate (ATP) at concentrations of 10–100 μM significantly enhanced lipopolysaccharide (LPS)-induced interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) release in stimulated whole blood. This effect was abrogated by P2X7 receptor antagonists, indicating P2X7-mediated signaling [1].
- NLRP3 inflammasome activation: In human gingival fibroblasts, ATP (5 mM) co-administered with Porphyromonas gingivalis LPS triggered NLRP3 inflammasome activation, as evidenced by caspase-1 cleavage and IL-1β secretion. This process was inhibited by doxycycline [3].

ATP (50 mg/kg; ip) protects the outer shell against infecting bacteria [4]. ATP covers IL-1β, KC and MIP-2 in the periphery as well as neutrophil recruitment [4].

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- Bacterial infection protection: Intraperitoneal injection of ATP (20 mg/kg) in mice significantly improved survival rates (60% vs. 20% in control) and reduced bacterial burden in a peritoneal Escherichia coli infection model. Protection was abolished in NLRP3 knockout mice, confirming NLRP3-dependent mechanism [4].

Human health is under constant threat of a wide variety of dangers, both self and nonself. The immune system is occupied with protecting the host against such dangers in order to preserve human health. For that purpose, the immune system is equipped with a diverse array of both cellular and non-cellular effectors that are in continuous communication with each other. The naturally occurring nucleotide adenosine 5'-triphosphate (ATP) and its metabolite adenosine (Ado) probably constitute an intrinsic part of this extensive immunological network through purinergic signaling by their cognate receptors, which are widely expressed throughout the body. This review provides a thorough overview of the effects of ATP and Ado on major immune cell types. The overwhelming evidence indicates that ATP and Ado are important endogenous signaling molecules in immunity and inflammation. Although the role of ATP and Ado during the course of inflammatory and immune responses in vivo appears to be extremely complex, we propose that their immunological role is both interdependent and multifaceted, meaning that the nature of their effects may shift from immunostimulatory to immunoregulatory or vice versa depending on extracellular concentrations as well as on expression patterns of purinergic receptors and ecto-enzymes. Purinergic signaling thus contributes to the fine-tuning of inflammatory and immune responses in such a way that the danger to the host is eliminated efficiently with minimal damage to healthy tissues[2].

In vitro studies suggest that extracellular nucleotides and nucleosides may be important regulators of inflammatory and immune responses. Most studies with adenosine 5'-triphosphate (ATP) have been performed in cell lines, which are remote from the human situation. The purpose of the present study was to determine the effects of ATP on TNF-alpha, IL-6 and IL-10 release in stimulated whole blood. Blood samples were drawn from healthy volunteers and incubated with ATP and lipopolysaccharide (LPS) + phytohemagglutinin (PHA) for 24 h. Contrary to expectations, ATP at 100 microM and 300 microM induced a reduction in TNF-alpha secretion by 32+/-8% (mean +/- SEM) and 65+/-4%, respectively. Furthermore, these ATP concentrations induced an increase in IL-10 secretion by 48+/-5% and 62+/-7% in whole blood. The ATP analogue adenosine 5'-O-(3-thiotriphosphate) (ATP-gamma-S) and adenosine 5'-diphosphate (ADP) also inhibited TNF-alpha release, but only ADP showed a stimulatory effect on IL-10. Co-treatment with adenosine deaminase did not reverse the ATP effect on TNF-alpha and IL-10. These results show, for the first time, that ATP inhibits the inflammatory response in stimulated whole blood as indicated by inhibition of TNF-alpha and stimulation of IL-10 release and that this effect is predominantly mediated by ATP and not by adenosine[1].

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- Whole blood stimulation: Human whole blood was incubated with LPS (1 μg/mL) and ATP (10–100 μM) for 6 hours. Cytokine levels in supernatants were measured by enzyme-linked immunosorbent assay (ELISA). P2X7 antagonist oxidized ATP (100 μM) blocked ATP-induced cytokine release [1].
- Gingival fibroblast stimulation: Human gingival fibroblasts were primed with LPS (1 μg/mL) for 3 hours, followed by ATP (5 mM) treatment for 30 minutes. Cell lysates were analyzed by Western blot for caspase-1 p20 and IL-1β, while supernatants were assayed for secreted IL-1β [3].

Animal/Disease Models: Fourweeks old Kunming mice (18-22 g) [4]
Doses: 50 mg/kg
Route of Administration: intraperitoneal (ip) injection before bacterial (E. coli) challenge
Experimental Results: Protect mice from bacterial infection.

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- Mouse infection model: ATP was dissolved in sterile saline and administered intraperitoneally (20 mg/kg) to C57BL/6 mice 1 hour before intraperitoneal injection of E. coli (1×10⁹ CFU). Survival was monitored for 72 hours, and peritoneal lavage fluid was cultured to quantify bacterial counts [4].

Metabolism / Metabolites
Organophosphate metabolism primarily occurs through oxidation, esterase hydrolysis, and reactions with glutathione. Demethylation and glucuronidation may also occur. Oxidation of organophosphate pesticides can produce moderately toxic products. Generally, thiophosphates themselves are not directly toxic and require oxidative metabolism to be converted into proximal toxins. Products produced by glutathione transferase reactions are generally less toxic. Paraoxygenase (PON1) is a key enzyme in organophosphate metabolism. PON1 can inactivate certain organophosphates through hydrolysis. PON1 hydrolyzes active metabolites in various organophosphate pesticides and nerve agents such as soman, sarin, and VX. The existence of PON1 polymorphism leads to differences in the enzyme level and catalytic efficiency of this esterase, which in turn suggests that different individuals may be more susceptible to the toxic effects of organophosphate exposure.

Toxicity Summary
ATP stores and transports chemical energy within cells. It also plays a crucial role in nucleic acid synthesis. ATP is produced through various cellular processes, most commonly in mitochondria, via oxidative phosphorylation catalyzed by ATP synthase. The total amount of ATP in the human body is approximately 0.1 moles. Human cells require the hydrolysis of 200 to 300 moles of ATP daily to meet their energy needs. This means that each ATP molecule is recycled 2000 to 3000 times per day. ATP cannot be stored, therefore its consumption must closely follow its synthesis. Toxicity Data
Oral LD50 (Rats): > 2 g/kg.

[1]. Immunoregulatory effects of adenosine 5'-triphosphate on cytokine release from stimulated whole blood. Eur J Immunol. 2005 Mar;35(3):852-8.

[2]. Adenosine 5'-triphosphate and adenosine as endogenous signaling molecules in immunity and inflammation. Pharmacol Ther. 2006 Nov;112(2):358-404.

[3]. Doxycycline inhibits NAcht Leucine-rich repeat Protein 3 inflammasome activation and interleukin-1β production induced by Porphyromonas gingivalis-lipopolysaccharide and adenosine triphosphate in human gingival fibroblasts. Arch Oral Biol. 2019 Nov;107:104514.

[4]. Adenosine-5'-Triphosphate (ATP) Protects Mice against Bacterial Infection by Activation of the NLRP3 Inflammasome. PLoS One. 2013; 8(5): e63759.

ATP is Adenanthin 5'-phosphate, where the 5'-phosphate group is a triphosphate group. It participates in the transport of chemical energy in metabolic pathways. ATP has multiple functions, including as a nutritional supplement, micronutrient, basic metabolite, and cofactor. It is an Adenanthin 5'-phosphate and purine ribonucleoside 5'-triphosphate. It is the conjugate acid of ATP (3-). Adenanthin triphosphate (ATP) is a metabolite found in or produced by Escherichia coli (K12 strain, MG1655 strain). ATP has also been reported in Jerusalem artichoke, Arabidopsis thaliana, and several other organisms with relevant data. ATP is an adenine nucleotide composed of three phosphate groups esterified onto a glycosyl group and is present in all living cells. ATP participates in energy production in metabolic processes and RNA synthesis. Furthermore, it functions as a neurotransmitter. In cancer research, ATP is synthesized to investigate its applications in weight loss and muscle strength enhancement. Adenanthin triphosphate (ATP) is a nucleotide consisting of a purine base (adenine) linked to the first carbon atom of a ribose sugar (a pentose sugar). Three phosphate groups are esterified at the fifth carbon atom of the ribose. During DNA replication and transcription, polymerases incorporate ATP into nucleic acids. ATP helps maintain cellular energy and participates in overall energy balance, thereby maintaining cellular homeostasis. ATP can mediate a wide variety of physiological processes, such as neurotransmission, inflammation, apoptosis, and bone remodeling, by interacting with specific purinergic receptors. Extracellular ATP and its metabolite Adenanthin have been shown to have multiple effects on almost all cell types in human skin. ATP appears to play a direct role in triggering inflammatory, regenerative, and fibrotic responses in the skin to mechanical injury, an indirect role in melanocyte proliferation and apoptosis, and a complex role in Langerhans cell-mediated adaptive immunity. During exercise, intracellular homeostasis depends on the matching of ATP supply and demand. Metabolites play a crucial role in transferring ATP demand to metabolic supply pathways. Stimulation of blood cells with extracellular ATP triggers various effects, including proliferation or differentiation, chemotaxis, release of cytokines or lysosomal components, and generation of reactive oxygen species or reactive nitrogen species. Many studies have observed elevated Adenanthin triphosphate (ATP) concentrations in erythrocytes of patients with chronic renal failure (CRF), but the mechanisms underlying these abnormalities remain controversial (A3367, A3368, A3369, A3370, A3371). Adenanthin triphosphate (ATP) is a metabolite found or produced in Saccharomyces cerevisiae. It is an adenine nucleotide with its glycosyl moiety esterified with three phosphate groups. Besides playing a crucial role in metabolism, ATP is also a neurotransmitter. Mechanism of action: ATP, as a damage-associated molecular model (DAMP), activates the P2X7 receptor, leading to potassium efflux and NLRP3 inflammasome assembly. This promotes caspase-1 activation and IL-1β/IL-18 secretion, thereby enhancing the host's defense against bacterial infection [2, 4]. - Immunomodulatory effects: Extracellular ATP can promote inflammation through the P2X7 signaling pathway or inhibit inflammation through an Adenanthin-mediated pathway, depending on local concentration and cellular environment [2].

C10H16N5O13P3

507.1810

506.995

C, 23.68; H, 3.18; N, 13.81; O, 41.01; P, 18.32

56-65-5

ATP disodium salt;987-65-5;ATP disodium trihydrate;51963-61-2;ATP dimagnesium;74804-12-9;ATP-13C10,15N5 disodium;ATP disodium salt hydrate;34369-07-8;ATP dipotassium;42373-41-1;ATP ditromethamine;102047-34-7;ATP-13C10,15N5;752972-20-6

5957

White to off-white solid powder

2.6±0.1 g/cm3

951.4±75.0 °C at 760 mmHg

187 - 190ºC (Decomposes)

529.2±37.1 °C

0.0±0.3 mmHg at 25°C

1.904

-4.18

7

17

8

31

800

4

C1=NC(=C2C(=N1)N(C=N2)[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)(O)OP(=O)(O)OP(=O)(O)O)O)O)N

ZKHQWZAMYRWXGA-KQYNXXCUSA-N

InChI=1S/C10H16N5O13P3/c11-8-5-9(13-2-12-8)15(3-14-5)10-7(17)6(16)4(26-10)1-25-30(21,22)28-31(23,24)27-29(18,19)20/h2-4,6-7,10,16-17H,1H2,(H,21,22)(H,23,24)(H2,11,12,13)(H2,18,19,20)/t4-,6-,7-,10-/m1/s1

[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate

Adenosine triphosphate; Ara-ATP; Atipi; Triphosphaden; Triphosphoric acid adenosine ester; Adenosine 5'-triphosphate; ATP; adenosine-5'-triphosphate; Myotriphos; Striadyne; Triadenyl;

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)

H2O : ≥ 100 mg/mL (~197.17 mM)

Solubility in Formulation 1: 100 mg/mL (197.17 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).

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