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].

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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]

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]

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]

[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.

Porcine atrial natriuretic peptide-(1-28) is a bioactive full-length atrial natriuretic peptide isolated from the pig heart. [1] This study found that Porcine atrial natriuretic peptide-(1-28) and brain natriuretic peptide (BNP) jointly inhibited the secretion of angiotensin II-stimulated endothelin-1 in vascular endothelial cells, and its mechanism of action may depend on cGMP. This is because cGMP analogues (8-bromo-cGMP) can mimic the action of ANP-(1-28), and the inhibitory effect of ANP-(1-28) is associated with an increase in intracellular cGMP levels. [1] This study suggests that ANP and BNP may act as physiological antagonists of the renin-angiotensin system, regulating vascular tone by modulating the secretion of the angiotensin-constricting peptide endothelin-1. [1]

C129H207N45O41S3

3140.49580216408

3139.469

1366000-58-9

Carperitide;89213-87-6

146160094

White to off-white solid powder

54

50

75

218

6970

0

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

NYSSIVMKYRVLPL-UHFFFAOYSA-N

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)

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

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)

H2O : ~50 mg/mL (~15.92 mM)

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.

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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).

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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)

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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).

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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

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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.

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