Endogenous Metabolite
Phosphocreatine disodium hydrate targets the creatine kinase enzyme system. It serves as a substrate for creatine kinase, which catalyzes the reversible transfer of a phosphate group between phosphocreatine and ADP to generate ATP and creatine. This system plays a critical role in maintaining ATP levels in tissues with high energy demands, such as muscle and brain. It does not target a specific receptor.

Research indicates that Phosphocreatine possesses neuroprotective and renoprotective potential. In terms of neuroprotection, Phosphocreatine (5-20 mM, 24 h) attenuates MGO (Methylglyoxal)-induced PC12 cell damage, inhibits apoptosis [3], and prevents the loss of mitochondrial membrane permeability [3]. Its mechanism of action involves normalizing mitochondrial function and reducing oxidative stress through the Akt-mediated Nrf2/HO-1 pathway [3]. Regarding renoprotection, Phosphocreatine (5-40 mM, 24 h) protects NRK-52E cells from MGO-induced kidney injury in a concentration-dependent manner [4], and its application at 10-40 mM for 4 h inhibits the production of renal oxidative stress metabolites [4].

---

In vitro, Phosphocreatine disodium hydrate is used as a substrate in enzymatic assays to measure creatine kinase activity. It is added to reaction mixtures containing creatine kinase, and the production of ATP is measured. It is also used in studies of cellular energy metabolism and as a supplement in cell culture media to support ATP production.

In the context of DOX-induced cardiotoxicity, Phosphocreatine treatment (200 mg/kg, i.p., every other day for 7 weeks) attenuated oxidative stress and apoptosis, and rescued myocardial tissue from necroptosis [2]. Additionally, in a diabetic nephropathy model, Phosphocreatine (20-40 mg/kg, i.v., daily for 6 weeks) demonstrated renoprotective effects by preserving renal function in Sprague-Dawley rats [4].

---

In vivo, Phosphocreatine disodium hydrate is used as a research tool to study cellular energy metabolism and ATP regeneration. It is a key component of the phosphocreatine shuttle, which transfers energy from mitochondria to sites of ATP consumption. It has been studied for its potential benefits in conditions involving energy depletion, such as ischemia and heart failure.

For non-cell-based assays, Phosphocreatine disodium hydrate is used as a substrate in creatine kinase activity assays. Enzyme activity is measured by monitoring the production of ATP or the consumption of phosphocreatine. The compound is incubated with creatine kinase and ADP, and the rate of ATP production is measured using coupled enzyme assays or by HPLC.

For in vitro cellular assays, cells are cultured in media containing Phosphocreatine disodium hydrate. The compound is taken up by cells and used to regenerate ATP. In studies of energy metabolism, cells are treated with the compound, and ATP levels are measured. The compound's ability to support cellular energy metabolism under stress conditions is assessed.

For in vivo animal studies, Phosphocreatine disodium hydrate can be administered to rodents via intraperitoneal or intravenous injection. In models of ischemia or heart failure, the compound's ability to maintain ATP levels and protect against tissue damage is assessed. In studies of muscle function, its effects on muscle contraction and fatigue are evaluated.

Phosphocreatine disodium hydrate has a molecular weight of 255.08 g/mol and a molecular formula of C4H8N3Na2O5P. It is a disodium salt hydrate. The compound is soluble in water. It should be stored at -20°C for long-term stability. It is for research use only and is not intended for human consumption.

The toxicity profile of Phosphocreatine disodium hydrate has not been extensively reported. As a naturally occurring compound, it is expected to have low toxicity. The compound is for research use only and is not intended for human consumption. Standard safety precautions should be observed when handling the compound.

[1]. Creatine: a dietary supplement and ergogenic aid. Nutr Rev. 1999 Feb;57(2):45-50.

[2]. Phosphocreatine attenuates doxorubicin-induced cardiotoxicity by inhibiting oxidative stress and activating TAK1 to promote myocardial survival in vivo and in vitro. Toxicology. 2021 Aug;460:152881.

[3]. Neuroprotective effect of phosphocreatine on oxidative stress and mitochondrial dysfunction induced apoptosis in vitro and in vivo: Involvement of dual PI3K/Akt and Nrf2/HO-1 pathways. Free Radic Biol Med. 2018 May 20;120:228-238.

[4]. Protection of diabetes-induced kidney injury by phosphocreatine via the regulation of ERK/Nrf2/HO-1 signaling pathway. Life Sci. 2020 Feb 1;242:117248.

Myocardial apoptosis and necroptosis are the main causes of doxorubicin (DOX)-induced cardiotoxicity and one of the important reasons limiting the clinical application of this drug. To date, the mechanism remains incompletely understood. The protective effect of creatine phosphate (PCr) in cardiac surgery and cardiology has been confirmed in numerous clinical trials. This study aimed to evaluate the protective effect of PCr against DOX-induced cardiotoxicity and explore its potential mechanism, particularly the transforming growth factor β-activated kinase 1 (TAK1)-mediated myocardial survival signaling pathway. Male Sprague-Dawley rats were used as an animal model, and patients were intraperitoneally injected with normal saline (NS), DOX (2 mg/kg), or DOX combined with PCr (200 mg/kg), respectively. Data showed that doxorubicin (DOX) significantly impaired cardiac function and structure, induced oxidative stress, Myocardial apoptosis and necroptosis, and significantly downregulated TAK1 expression levels; while intervention with creatine phosphate (PCr) significantly alleviated cardiac dysfunction, oxidative stress, Myocardial apoptosis and necroptosis, especially mitigating the decrease in TAK1 expression. In in vitro experiments, H9c2 cells were pretreated with PCr (0.5 mM), N-acetyl-L-cysteine (NAC, 0.5 mM), or 5Z-7-oxozeenol (5Z-7-Ox, 1 μM) for 1 hour, followed by DOX (1 μM) treatment for 24 hours. The results showed that inhibiting TAK1 further exacerbated DOX-induced apoptosis and necrotizing apoptosis in H9c2 cells, but had no effect on oxidative stress. PCr or NAC pretreatment enhanced antioxidant activity, reduced oxidative stress, and significantly alleviated DOX-induced apoptosis and necrotizing cell death in H9c2 cells. Consistent with in vivo results, PCr or NAC significantly inhibited DOX-induced decrease in TAK1 expression. In summary, DOX-induced oxidative stress inhibits TAK1 expression, leading to Myocardial apoptosis and necrotic cell death; while PCr intervention can enhance antioxidant activity, reduce oxidative stress, thereby activating the TAK1 signaling pathway, promoting Myocardial survival, and ultimately reducing DOX-induced cardiotoxicity. [2]

---

Phosphocreatine disodium hydrate (CAS 19333-65-4) is the disodium salt hydrate form of phosphocreatine, a phosphorylated form of creatine that serves as an energy reserve in muscle and brain tissue. It is used as a substrate in creatine kinase assays and in studies of cellular energy metabolism. It is available for research purposes only.

C4H10N3NA2O6P

273.09200334549

255

19333-65-4

Phosphocreatine disodium;922-32-7;Phosphocreatine;67-07-2;Sodium creatine phosphate dibasic tetrahydrate;71519-72-7;Phosphocreatine dipotassium;18838-38-5;67-07-2;19333-65-4;71519-72-7;922-32-7;

44134639

White to off-white solid powder

6

10

3

19

259

0

P(N=C(N)N(C)CC(=O)O)(=O)([O-])[O-].[Na+].[Na+].O

HUWYWJSJJDCZRQ-UHFFFAOYSA-L

InChI=1S/C4H10N3O5P.2Na.4H2O/c1-7(2-3(8)9)4(5)6-13(10,11)12;;;;;;/h2H2,1H3,(H,8,9)(H4,5,6,10,11,12);;;4*1H2/q;2*+1;;;;/p-2

disodium;2-[methyl-(N'-phosphonatocarbamimidoyl)amino]acetic acid;tetrahydrate

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

---

Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

View More

Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

View More

Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

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