 (1-28), human, porcine Acetate Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C129H207N45O41S3 |
| Molecular Weight | 3140.49580216408 |
| Exact Mass | 3139.469 |
| CAS # | 1366000-58-9 |
| Related CAS # | Carperitide;89213-87-6 |
| PubChem CID | 146160094 |
| Appearance | White to off-white solid powder |
| Hydrogen Bond Donor Count | 54 |
| Hydrogen Bond Acceptor Count | 50 |
| Rotatable Bond Count | 75 |
| Heavy Atom Count | 218 |
| Complexity | 6970 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | C(=O)(O)C.[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)NC([C@H]1NC(CNC([C@@H](NC(CNC([C@@H](NC([C@@H](NC([C@@H](NC(CNC([C@@]([H])(NC(=O)C(CCCNC(N)=N)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@@H](CCCNC(N)=N)NC(=O)CNC(=O)CNC(=O)[C@]([H])(CC2C=CC=CC=2)NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CO)CSSC1)[C@@H](C)CC)=O)=O)C)=O)CCC(=O)N)=O)CO)=O)=O)CC(C)C)=O)=O)=O)(C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(=O)O)CC1C=CC(O)=CC=1)CC1C=CC=CC=1 |
| InChi Key | NYSSIVMKYRVLPL-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C127H203N45O39S3.C2H4O2/c1-9-64(6)99-121(209)150-52-94(182)151-65(7)100(188)155-76(34-35-91(129)179)109(197)167-85(56-174)104(192)149-53-96(184)153-78(43-62(2)3)102(190)148-54-97(185)154-89(119(207)164-82(48-92(130)180)114(202)169-86(57-175)116(204)163-81(46-67-23-14-11-15-24-67)113(201)158-73(27-18-39-143-125(135)136)107(195)166-84(122(210)211)47-68-30-32-69(178)33-31-68)60-213-214-61-90(171-118(206)88(59-177)170-117(205)87(58-176)168-108(196)74(28-19-40-144-126(137)138)156-106(194)72(26-17-38-142-124(133)134)157-112(200)79(44-63(4)5)161-101(189)70(128)55-173)120(208)162-80(45-66-21-12-10-13-22-66)103(191)147-50-93(181)146-51-95(183)152-71(25-16-37-141-123(131)132)105(193)160-77(36-42-212-8)110(198)165-83(49-98(186)187)115(203)159-75(111(199)172-99)29-20-41-145-127(139)140;1-2(3)4/h10-15,21-24,30-33,62-65,70-90,99,173-178H,9,16-20,25-29,34-61,128H2,1-8H3,(H2,129,179)(H2,130,180)(H,146,181)(H,147,191)(H,148,190)(H,149,192)(H,150,209)(H,151,182)(H,152,183)(H,153,184)(H,154,185)(H,155,188)(H,156,194)(H,157,200)(H,158,201)(H,159,203)(H,160,193)(H,161,189)(H,162,208)(H,163,204)(H,164,207)(H,165,198)(H,166,195)(H,167,197)(H,168,196)(H,169,202)(H,170,205)(H,171,206)(H,172,199)(H,186,187)(H,210,211)(H4,131,132,141)(H4,133,134,142)(H4,135,136,143)(H4,137,138,144)(H4,139,140,145);1H3,(H,3,4) |
| Chemical Name | acetic acid;2-[[2-[[2-[[2-[[4-amino-2-[[52-[[2-[[2-[[2-[[2-[[2-[(2-amino-3-hydroxypropanoyl)amino]-4-methylpentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-19-(3-amino-3-oxopropyl)-49-benzyl-28-butan-2-yl-31,40-bis(3-carbamimidamidopropyl)-34-(carboxymethyl)-16-(hydroxymethyl)-22-methyl-10-(2-methylpropyl)-37-(2-methylsulfanylethyl)-6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51-hexadecaoxo-1,2-dithia-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50-hexadecazacyclotripentacontane-4-carbonyl]amino]-4-oxobutanoyl]amino]-3-hydroxypropanoyl]amino]-3-phenylpropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(4-hydroxyphenyl)propanoic acid |
| HS Tariff Code | 2934.99.9001 |
| Storage |
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. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| ln Vitro |
ANP is a diuretic, natriuretic, and vasodilatory peptide hormone derived from the mammalian heart. In cultivated swine endothelial cells, pig ANP (1-28) inhibited immunoreactive endothelin-1 release while increasing cellular cGMP levels after angiotensin II stimulation. pig ANP (1-28) reduces immunoreactive endothelin-1 production in the pig aorta following Ang II stimulation [1]. ANP is a cardiac hormone that regulates electrolyte and fluid balance. ANP inhibits angiotensin II (ANGII) and thrombin-stimulated endothelin-1 production from cultured human umbilical vein endothelial cells. Human ANP (1-28) suppresses ir-endothelin-1 secretion while increasing cyclic GMP in human umbilical vein endothelial cells [2]. In normal rat glomeruli, human 125I-ANP (1-28) binds to a single high-affinity receptor population with an average equilibrium dissociation constant of 0.46 nM. Human ANP (1-28) binds to glomerular ANP receptors with high affinity, stimulating cGMP buildup. Human ANP (1–28) promotes cGMP synthesis but not cAMP production in normal rat glomeruli [3]. In porcine aortic strips with intact endothelium, porcine ANP-(1-28) dose-dependently inhibited angiotensin II (Ang II, 10^-8 M)-stimulated immunoreactive endothelin-1 secretion during a 4-hour incubation at 37°C. At concentrations of 10^-8 M and 10^-6 M, porcine ANP-(1-28) significantly reduced the secreted endothelin-1 levels compared to Ang II stimulation alone. It did not significantly affect spontaneous (basal) immunoreactive endothelin-1 secretion at these concentrations. [1] In cultured porcine aortic endothelial cells, porcine ANP-(1-28) (10^-10 to 10^-6 M) dose-dependently inhibited immunoreactive endothelin-1 secretion stimulated by Ang II (10^-8 M) during a 4-hour incubation. This inhibitory effect was paralleled by a dose-dependent increase in cellular cyclic GMP (cGMP) levels when cells were co-incubated with ANP and the phosphodiesterase inhibitor IBMX for 30 minutes. A negative correlation was observed between the percent decrease in immunoreactive endothelin-1 secretion and the percent increase in cellular cGMP. [1] In competitive radioligand binding assays using cultured porcine endothelial cells, porcine ANP-(1-28) at 10^-6 M did not affect the binding of 125I-labeled Ang II to its receptor. Saturation binding studies and Scatchard analysis showed that porcine ANP-(1-28) (10^-6 M) did not alter the dissociation constant (Kd) or the maximum number of binding sites (Bmax) for Ang II. [1] |
| Enzyme Assay | Cellular cGMP levels were measured in cultured porcine endothelial cells to assess the activation of the ANP receptor/guanylyl cyclase pathway. After pre-washing, cell monolayers were stimulated for 30 minutes at 37°C with different concentrations of porcine ANP-(1-28) dissolved in serum-free culture medium containing 0.5 mM 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor). The reaction was stopped by rapid aspiration and addition of ice-cold 65% ethanol. The cGMP content in the cell extracts was then quantified using a commercial cGMP assay kit according to the manufacturer's instructions. [1] |
| Cell Assay |
For measuring Ang II-stimulated endothelin-1 secretion in cultured porcine endothelial cells, confluent cells (passages 3-6) in tissue culture flasks were washed twice with serum-free medium. The cells were then incubated for 4 hours at 37°C in 2 ml of serum-free medium containing Ang II (10^-8 M) with or without various concentrations of porcine ANP-(1-28). After incubation, the medium was collected, centrifuged, and stored for subsequent radioimmunoassay of immunoreactive endothelin-1. [1] For the radioligand binding assay to assess ANP's effect on Ang II receptors, confluent porcine endothelial cells grown in multi-well plates were used. Cells were washed and then incubated at 37°C with a buffer containing 5 x 10^-10 M 125I-labeled Ang II in the presence or absence of 10^-6 M porcine ANP-(1-28) for 15 minutes to reach equilibrium. After incubation, cells were washed with ice-cold buffer to remove unbound ligand. To distinguish surface-bound ligands, cells were treated with 0.2 M acetic acid (pH 2.5) containing 0.5 M NaCl for 6 minutes at 4°C. The extracted radioactivity was counted to determine specific binding. Non-specific binding was determined in the presence of 10^-5 M unlabeled Ang II. [1] |
| References |
[1]. Atrial and brain natriuretic peptides inhibit the endothelin-1 secretory response to angiotensin II in porcine aorta. Circ Res. 1992 Feb;70(2):241-7. [2]. Inhibition by atrial and brain natriuretic peptides of endothelin-1 secretion after stimulation with angiotensin II and thrombin of cultured human endothelial cells. J Clin Invest. 1991 Jun;87(6):1999-2004. [3]. Physiologic regulation of atrial natriuretic peptide receptors in rat renal glomeruli. J Clin Invest. 1985 Dec;76(6):2049-56. |
| Additional Infomation |
Porcine ANP-(1-28) is the full-length, biologically active form of atrial natriuretic peptide isolated from porcine heart. [1] In this study, porcine ANP-(1-28), along with brain natriuretic peptide (BNP), was found to inhibit angiotensin II-stimulated endothelin-1 secretion from vascular endothelium, likely through a cGMP-dependent mechanism, as its effects were mimicked by a cGMP analogue (8-bromo-cGMP) and correlated with increases in intracellular cGMP levels. [1] The study suggests that ANP and BNP may act as physiological antagonists to the renin-angiotensin system in regulating vascular tone, not only through direct vasodilatory effects but also by modulating the secretion of the vasoconstrictor peptide endothelin-1. [1] |
Solubility Data
| Solubility (In Vitro) | H2O : ~50 mg/mL (~15.92 mM) |
| Solubility (In Vivo) |
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 Formulations (e.g. IP/IV/IM/SC) 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). 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 Formulations 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). 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.3184 mL | 1.5921 mL | 3.1842 mL | |
| 5 mM | 0.0637 mL | 0.3184 mL | 0.6368 mL | |
| 10 mM | 0.0318 mL | 0.1592 mL | 0.3184 mL |