Spermine NONOate

Spermine NONOate is a diazeniumdiolate nitric oxide donor [1].

Nitric oxide (NO) increases intracellular guanosine 3' 5' cyclic monophosphate and activates guanylate cyclase, which then cause relaxation of vascular smooth muscle [1].

Spermine NONOate is a diazeniumdiolate nitric oxide donor. In rat pulmonary artery rings, spermine NONOate (100 μM) increased guanosine 3'5' cyclic monophosphate (cyclic GMP) by 35 fold and caused cyclic GMP-independent relaxation by activation of sarco-endoplasmic reticulum Ca2+-ATPase, Na+/K+-ATPase and calcium-activated potassium channels [1]. Spermine NONOate inhibited human low-density lipoprotein (LDL) oxidation induced by the copper(II) sulfate [2].

In rats with bilateral uterine horn injury, spermine NONOate (1 mg) inhibited the formation of adhesions. Also, The N,O-carboxymethylchitosan (NOCC) plus spermine NONOate group had a lower severity score and extent score than the spermine NONOate group. These results suggested that spermine NONOate reduced the postoperative adhesions [3].

References:
[1].  Homer KL, Wanstall JC. Cyclic GMP-independent relaxation of rat pulmonary artery by spermine NONOate, a diazeniumdiolate nitric oxide donor. Br J Pharmacol, 2000, 131(4): 673-682.
[2].  Goss SP, Hogg N, Kalyanaraman B. The antioxidant effect of spermine NONOate in human low-density lipoprotein. Chem Res Toxicol, 1995, 8(5): 800-806.
[3].  Duran B, Ak D, Cetin A, et al. Reduction of postoperative adhesions by N,O-carboxymethylchitosan and spermine NONOate in rats. Exp Anim, 2003, 52(4): 267-272.

A comparative study of NONOate based NO donors: spermine NONOate is the best suited NO donor for angiogenesis.[Pubmed:24333563]

Nitric Oxide. 2014 Jan 30;36:76-86.

Nitric oxide (NO) is a known modulator of angiogenesis. The NONOate subfamily of NO donors has long been used in experimental and clinical studies to promote angiogenesis. However, no studies have been conducted yet to compare the angiogenesis potential of these NO donors in respect to their pattern of NO release. We hypothesize that having different pattern of NO release, each of the NO donors in NONOate subfamily can promote key stages of angiogenesis in differential manner. To verify our hypothesis, NO donors with half life ranging from seconds to several hours and having very different pattern of NO release were selected to evaluate their efficacy in modulating angiogenesis. Endothelial tube formation using EAhy926 cells was maximally increased by Spermine NONOate (SP) treatment. SP treatment maximally induced both ex vivo and in vivo angiogenesis using egg yolk and cotton plug angiogenesis models respectively. Experiment using chick embryo partial ischemia model revealed SP as the best suited NO donor to recover ischemia driven hampered angiogenesis. The present study elaborated that differential release pattern of NO by different NO donors can modulate angiogenesis differentially and also suggested that SP have a unique pattern of NO release that best fits for angiogenesis.

Effects of spermine NONOate and ATP on protein aggregation: light scattering evidences.[Pubmed:23289636]

BMC Biophys. 2013 Jan 4;6:1.

UNLABELLED: BACKGROUND AND OBJECTIVE: Regulating protein function in the cell by small molecules, provide a rapid, reversible and tunable tool of metabolic control. However, due to its complexity the issue is poorly studied so far. The effects of small solutes on protein behavior can be studied by examining changes of protein secondary structure, in its hydrodynamic radius as well as its thermal aggregation. The study aim was to investigate effects of adenosine-5'-triphosphate (ATP), Spermine NONOate (NO donor) as well as sodium/potassium ions on thermal aggregation of albumin and hemoglobin. To follow aggregation of the proteins, their diffusion coefficients were measured by quasi-elastic light scattering (QELS) at constant pH (7.4) in the presence of solutes over a temperature range from 25 degrees C to 80 degrees C. RESULTS AND DISCUSSION: 1) Spermine NONOate persistently decreased the hemoglobin aggregation temperature Tairrespectively of the Na+/K+ environment, 2) ATP alone had no effect on the protein's thermal stability but it facilitated protein's destabilization in the presence of Spermine NONOate and 3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. CONCLUSION: The ATP effect on protein aggregation was ambiguous: ATP alone had no effect on the protein's thermal stability but it facilitated protein's destabilization in the presence of nitric oxide. The magnitude and direction of the observed effects strongly depended on concentrations of K+ and Na+ in the solution.

Effects of spermine NONOate and ATP on the thermal stability of hemoglobin.[Pubmed:22929146]

BMC Biophys. 2012 Aug 28;5:16.

BACKGROUND: Minor changes in protein structure induced by small organic and inorganic molecules can result in significant metabolic effects. The effects can be even more profound if the molecular players are chemically active and present in the cell in considerable amounts. The aim of our study was to investigate effects of a nitric oxide donor (Spermine NONOate), ATP and sodium/potassium environment on the dynamics of thermal unfolding of human hemoglobin (Hb). The effect of these molecules was examined by means of circular dichroism spectrometry (CD) in the temperature range between 25 degrees C and 70 degrees C. The alpha-helical content of buffered hemoglobin samples (0.1 mg/ml) was estimated via ellipticity change measurements at a heating rate of 1 degrees C/min. RESULTS: Major results were: 1) Spermine NONOate persistently decreased the hemoglobin unfolding temperature Tuirrespectively of the Na + /K + environment, 2) ATP instead increased the unfolding temperature by 3 degrees C in both sodium-based and potassium-based buffers and 3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. Moreover, the presence of potassium facilitated a partial unfolding of alpha-helical structures even at room temperature. CONCLUSION: The obtained data might shed more light on molecular mechanisms and biophysics involved in the regulation of protein activity by small solutes in the cell.

Pharmacological profile of a nitric oxide donor spermine NONOate in the mouse corpus cavernosum.[Pubmed:25551923]

Turk J Med Sci. 2014;44(4):569-75.

BACKGROUND/AIM: To investigate the effects of Spermine NONOate in the cavernous tissue obtained from mice treated or untreated with sildenafil. MATERIALS AND METHODS: We studied the effects of Spermine NONOate on the tone and nitrergic relaxation responses of isolated mouse corpus cavernosum and compared them with sodium nitroprusside in the absence or presence of L-nitroarginine, hydroxocobalamin, pyrogallol, diethyldithiocarbamate (DETCA), or 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). The neurogenic contractions and relaxations of the tissues were induced by electrical field stimulation. Some mice received a single oral dose of sildenafil and after 1 h the effects of Spermine NONOate were evaluated by in vitro studies. RESULTS: Spermine NONOate relaxed mouse corpus cavernosum in a concentration-dependent manner. Spermine-NONOate-induced relaxation was relatively slow to develop and it was reversible and reproducible. These relaxations were significantly suppressed by hydroxocobalamin, diethyldithiocarbamate, or ODQ, but not by L-nitroarginine or pyrogallol. Spermine NONOate potentiated the nitrergic relaxations to electrical field stimulation (EFS), whereas it significantly reduced EFS-induced contractions. Sildenafil treatment can enhance the relaxant responses to Spermine NONOate and EFS. CONCLUSION: These findings suggest that Spermine NONOate has a potent relaxant action in cavernous tissue and this effect can be potentiated by oral sildenafil treatments. Spermine NONOate may be considered an attractive treatment for erectile dysfunction in pathologic disorders with a lack of endogenous NO production.

Comparative effects of several nitric oxide donors on intracellular cyclic GMP levels in bovine chromaffin cells: correlation with nitric oxide production.[Pubmed:10401570]

Br J Pharmacol. 1999 Jun;127(3):779-87.

1. Sodium nitroprusside, S-nitroso-N-acetyl-D,L-penicillamine, Spermine NONOate and DEA NONOate raised cyclic GMP levels in bovine chromaffin cells in a time and concentration dependent manner with different potencies, the most potent being DEA/NO with an EC50 value of 0.38 +/- 0.02 microM. 2. Measurements of NO released from these donors revealed that DEA/NO decomposed with a half-life (t1/2) of 3.9 +/- 0.2 min. The t1/2 for SPER/NO was 37 +/- 3 min. SNAP decomposed more slowly (t1/2 = 37 +/- 4 h) and after 60 min the amount of NO produced corresponded to less than 2% of the total SNAP present. The rate of NO production from SNAP was increased by the presence of glutathione. 3. For DEA/NO and SPER/NO there was a clear correlation between nitric oxide production and cyclic GMP increases. Their threshold concentrations were 0.05 microM and maximal effective concentration between 2.5 and 5 microM. 4. For SNAP, threshold activation was seen at 1 microM, whereas full activation required a higher concentration (500-750 microM). The dose-response for SNAP increases in cyclic GMP was shifted nearly two orders of magnitude lower in the presence of glutathione. At higher concentrations an inhibition of cyclic GMP accumulation was found. This effect was not observed with either the nitric oxide-deficient SNAP analogue or other NO donors. 5. Although NO-donors are likely to be valuable for studying NO functions, their effective concentrations and the amount of NO released by them are very different and should be assessed in each system to ensure that physiological concentrations of NO are used.

In vitro comparison of two NONOates (novel nitric oxide donors) on rat pulmonary arteries.[Pubmed:9761423]

Eur J Pharmacol. 1998 Aug 28;356(1):49-57.

The pulmonary vasorelaxant properties of two NONOates (diazeniumdiolates) were examined because this novel group of nitric oxide (NO) donors may be useful in pulmonary hypertension. MAHMA NONOate ((Z)-1- inverted question markN-Methyl-N-[6-(N-methylammoniohexyl)amino] inverted question mark diazen-1-ium-1,2-diolate) and Spermine NONOate ((Z)- 1- inverted question markN-[3-aminopropyl]-N-[4-(3-aminopropylammonio)butyl]-amino inverted question markdi azen-1-ium-1,2-diolate) decomposed at different rates (half-lives 1.3 min and 73 min, respectively; 37 degrees C, pH 7.3) but generated the same total amount of NO. They fully relaxed submaximally contracted ring preparations of main and intralobar pulmonary arteries from rats. Responses were inhibited by the guanylate cyclase inhibitor, ODQ (1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one). Potency was not affected by choice of contractile spasmogen (phenylephrine, endothelin-1, thromboxane-mimetic) or endothelium removal, and tolerance did not develop; thus the drugs had properties important for use in pulmonary hypertension. MAHMA NONOate was 10-40-fold more potent than Spermine NONOate but responses to Spermine NONOate were more sustained (Spermine NONOate > 60 min; MAHMA NONOate < 7 min). It is concluded that the differences in potency and time-course reflect the different rates of NO generation by these NONOates.

Measurement and modeling of nitric oxide release rates for nitric oxide donors.[Pubmed:9114977]

Chem Res Toxicol. 1997 Apr;10(4):408-13.

An accurate model of the nitric oxide (NO)-release rate is essential for predicting the temporal NO-release rate and resulting NO concentrations for NO donors. Knowledge of the NO-release rate and/or the NO concentration is beneficial for assessing the physiological or pathological effects of NO on cell systems. This study describes a method to measure the temporal NO-release rate from NO donor compounds utilizing a modified ultrafiltration cell. For this study, the NO-release rates of Spermine NONOate and diethylamine NONOate were measured and kinetically modeled at pH 7.4 and 37 degrees C. An advantage of this method is that complete dissolution of the NONOate was not necessary for modeling the NO-release rate. One model parameter, which is the number of moles of NO released per mole of decomposed NONOate, is 1.7 +/- 0.1 and 1.5 +/- 0.2 for spermine and diethylamine NONOate, respectively. The other model parameter, which is the NONOate first-order decomposition rate constant, is 0.019 +/- 0.002 min-1 and 0.47 +/- 0.10 min-1 for spermine and diethylamine NONOate, respectively, as determined from NO concentration profiles. The decomposition rate constant measured by spectrophotometry was consistent with the above value for Spermine NONOate but was approximately half the above value for diethylamine NONOate. Preliminary experiments using spectrophotometry showed that for both NONOates the decomposition activation energy was approximately 100 kJ/mol. The NO-release rate model for Spermine NONOate was applied to a model for predicting the NO concentration in oxygenated solution. The NO concentration was measured in phosphate buffer, culture medium, and Tyrode's solution. Excellent agreement was observed between experimental and predicted NO concentrations.

Complexes of .NO with nucleophiles as agents for the controlled biological release of nitric oxide. Vasorelaxant effects.[Pubmed:1956043]

J Med Chem. 1991 Nov;34(11):3242-7.

Selected nucleophile/nitric oxide adducts [compounds which contain the anionic moiety, XN(O-)N = O] were studied for their ability to release nitric oxide spontaneously in aqueous solution and for possible vasoactivity. The diversity of structures chosen included those in which the nucleophile residue, X, was that of a secondary amine [Et2N, as in [Et2NN(N = O)O]Na, 1], a primary amine [iPrHN, as in [iPrHNN(N = O)O]Na, 2], a polyamine, spermine [as in the zwitterion H2N(CH2)3NH2+(CH2)4N[N(N = O)O-](CH2)3NH2, 3], oxide [as in Na[ON(N = O)O]Na, 4], and sulfite [as in NH4[O3SN(N = O)O]NH4, 5]. The rate constants (k) for decomposition in pH 7.4 phosphate buffer at 37 degrees C, as measured by following loss of chromophore at 230-260 nm, were as follows: 1, 5.4 x 10(-3) s-1; 2, 5.1 x 10(-3) s-1; 3, 0.30 x 10(-3) s-1; 4, 5.0 x 10(-3) s-1; and 5, 1.7 x 10(-3) s-1. The corresponding extents of nitric oxide release (ENO) were 1.5, 0.73, 1.9, 0.54, and 0.001 mol/mol of starting material consumed, respectively, as determined from the integrated chemiluminescence response. Vasodilatory activities expressed as the concentrations required to induce 50% relaxation in norepinephrine-constricted aortic rings bathed in pH 7.4 buffer at 37 degrees C (EC50) were as follows: 1, 0.19 microM; 2, 0.45 microM; 3, 6.2 microM; 4, 0.59 microM; and 5, 62 microM. Vasorelaxant potency (expressed as 1/EC50) was strongly correlated with the quantity of .NO calculated from the physicochemical data to be released in the interval required to achieve maximum relaxation at the EC50 doses (r = 0.995). This suggests that such nucleophile/.NO adducts might generally be useful as vehicles for the nonenzymatic generation of nitric oxide, in predictable amounts and at predictable rates, for biological purposes. The particular significance for possible drug design is underscored in the very favorable potency comparison between several of these agents and the established nitrovasodilators sodium nitroprusside and glyceryl trinitrate (EC50 values of 2.0 and greater than 10 microM, respectively) in parallel aortic ring tests.