Dataset on structure, stability and myocardial effects of a new hybrid aspirin containing nitrogen monoxide-releasing molsidomine moiety.

Herein 1H and 13C NMR spectra of ERJ-500, a new hybrid aspirin derivative, covalently conjugated to nitrogen monoxide donor linsidomine are presented as well as NMR spectra of its synthetic intermediate compounds. HPLC-MS measurements data are also included, demonstrating the stability of the linsidomine-aspirin hybrid in oxidation reactions. This data article also concerns miscellaneous myocardial parameters of isolated rat hearts as a complementation of the tables shown in the paper entitled "A new, vasoactive hybrid aspirin containing nitrogen monoxide-releasing molsidomine moiety" Szoke et al., 2019. Column tables represent data of aorta flow, aortic pressure, derivated aortic pressure and cardiac output.

Fig. 2

1H NMR spectra of compound 5.

Data

Spectra from 1H and 13C NMR measurements are reported to prove the structure of the synthesized compounds [1] (see Fig. 1, Fig. 2, Fig. 4, Fig. 5, Fig. 6, Fig. 7, Fig. 8, Fig. 9). HPLC-MS measurements were used to study the oxidative stability of the compound ERJ-500, representative total ion chromatogram of the oxidation by synthetic porphyrin and the Chemical Fenton System can be seen (Fig. 10, Fig. 11). Cardiac parameters (aorta flow, aortic pressure, derivated aortic pressure, and cardiac output) were registered (Fig. 12) in “isolated working hearts” treated with ERJ-500.

Fig. 1

Structure of ERJ-500.

Fig. 4

1H NMR spectra of compound 6.

Fig. 5

13C NMR spectra of compound 6.

Fig. 6

1H NMR spectra of compound 7.

Fig. 7

13C NMR spectra of compound 7.

Fig. 8

1H NMR spectra of compound ERJ-500.

Fig. 9

13C NMR spectra of compound ERJ-500.

Fig. 10

Representative total ion chromatogram of the oxidation by synthetic porphyrin. On the control chromatogram (red) the peak at 6.59 represents ERJ-500. After oxidation (green chromatogram) the peak of ERJ-500 at 6.58 remained unchanged.

Fig. 11

Representative total ion chromatogram of the oxidation by the Chemical Fenton System. On the control chromatogram (red) the peak at 6.54 represents ERJ-500. After oxidation (green chromatogram) the peak of ERJ-500 at 6.53 remained unchanged.

Fig. 12

Myocardial function. The results show aorta flow, aortic pressure, derivated aortic pressure, and cardiac output in control and ERJ-500 treated hearts.

13C NMR spectra of compound 5.

Characterization of new compounds

The 1H NMR (400 MHz) and 13C NMR (101 MHz) spectra were recorded with a Bruker DRX-400 spectrometer at 25 °C. Chemical shifts are referenced to Me4Si (0.00 ppm for 1H) and to the residual solvent signals (CDCl3: 77.1 for 13C).

Compound 5

1H NMR (400 MHz, CDCl3): δ 8.03 (dd, J = 7.8 Hz, J = 1.8 Hz, 1H, arom), 7.53 (td, J = 7.8 Hz, J = 1.8 Hz, 1H, arom), 7.47–7.45 (m, 6H, arom), 7.30–7.19 (m, 10H, arom), 7.08 (dd, J = 8.1 Hz, J = 0.8 Hz, 1H), 4.41–4.39 (m, 2H, TEG-CH2), 3.78–3.76 (m, 2H, TEG-CH2), 3.70–3.65 (m, 10H, 5 × TEG-CH2), 3.23 (t, J = 5.2 Hz, 2H, TEG-CH2), 2.34 (s, 3H, CH3 Ac); 13C NMR (101 MHz, CDCl3): δ 169.9 (1C, Cq Ac), 164.5 (1C, COO), 150.8 (1C, Cq arom), 144.2 (3C, Cq arom), 134.0, 132.0, 128.8, 127.9, 127.0, 126.1, 123.9 (19C, arom), 123.3 (1C, Cq arom), 86.6 (1C, Cq Tr), 70.9, 70.8, 70.7, 69.2, 64.4, 63.4 (8C, 8 × TEG-CH2), 21.1 (1C, CH3 Ac).

Compound 6

1H NMR (400 MHz, CDCl3): δ 8.05 (dd, J = 7.8 Hz, J = 1.6 Hz, 1H, arom), 7.56 (td, J = 7.9 Hz, J = 1.6 Hz, 1H, arom), 7.32 (td, J = 7.6 Hz, J = 1.2 Hz, 1H, arom), 7.11 (dd, J = 8.1 Hz, J = 1.2 Hz, 1H, arom), 4.45–4.43 (m, 2H, TEG-CH2), 3.81–3.78 (m, 2H, TEG-CH2), 3.74–3.65 (m, 10H, 5 × TEG-CH2), 3.60–3.58 (m, 2H, TEG-CH2), 2.62 (s, 1H, TEG-OH), 2.36 (s, 3H, CH3 Ac); 13C NMR (101 MHz, CDCl3): δ 169.9 (1C, Cq COO), 164.5 (1C, Cq Ac), 150.8 (1C, Cq arom), 134.1, 132.0, 126.1, 123.9 (4C, arom), 123.2 (1C, Cq arom), 72.5, 70.8, 70.7, 70.6, 70.4, 69.2, 64.3, 61.8 (8C, 8 × TEG-CH2), 21.1 (1C, CH3 Ac).

Compound 7

1H NMR (400 MHz, CDCl3): δ 8.41 (s, 1H, CH sydnone), 8.27 (d, J = 9.1 Hz, 2H, arom), 7.42 (d, J = 9.1 Hz, 1H, arom), 3.87–3.85 (m, 2H, CH2 morpholine), 3.64–3.61 (m, 2H, CH2 morpholine); 13C NMR (101 MHz, CD3OD): δ 183.5 (1C, Cq carbamate), 167.0 (1C, Cq sydnone), 134.5, 132.1, (4C, arom), 125.3 (1C, CH), 74.3, 62.9 (4C, 4 × morfoline-CH2).

Compound ERJ-500

1H NMR (400 MHz, CDCl3): δ 8.03 (dd, J = 7.9 Hz, J = 1.7 Hz, 1H, arom), 7.70 (s, 1H, CH sydnone), 7.56 (ddd, J = 8.1, 7.4, 1.8 Hz, 1H, arom), 7.31 (td, J = 7.7 Hz, 1.1 Hz, 1H, arom), 7.10 (dd, J = 8.1 Hz, J = 1.1 Hz, 1H, arom), 4.43–4.41 (m, 2H, CH2 morpholine), 4.26–4.24 (m, 2H, CH2 morpholine), 3.94–3.92 (m, 4H, 2 × TEG-CH2), 3.80–3.78 (m, 2H, CH2 morpholine), 3.74–3.72 (m, 2H, CH2 morpholine), 3.68–3.63 (m, 8H, 4 × TEG-CH2), 3.51–3.49 (m, 4H, 2 × TEG-CH2), 2.35 (s, 3H, CH3 Ac); 13C NMR (101 MHz, CDCl3): δ 174.2 (1C, Cq carbamate), 169.7 (1C, Cq COO), 164.4 (1C, Cq Ac), 161.2 (1C, Cq sydnone), 150.6 (1C, Cq arom), 133.8, 131.8, 125.9, 123.7 (4C, arom), 123.2 (1C, Cq arom), 70.6, 70.5, 69.3, 69.0, 65.4, 64.6, 64.3, 54.6 (13C, 1 × sydnone-C, 4 × morpholine-CH2, 8 × TEG-CH2), 20.9 (1C, CH3 Ac).

Representative chromatograms of oxidative stability assays

Experimental design, materials and methods

LC-MS measurements

The reaction mixture was analyzed with an LTQ-XL linear ion trap mass spectrometer coupled with the Accela LC system (Thermo Fisher Scientific, Waltham, MA, USA). The HPLC separation was performed using a Kinetex XB-C18 2.6 μm column, 0.1% formic acid in water, and ACN with 0.1% formic acid with gradient elution, and the flow rate was set to 300 μL/min. The method parameters for mass spectrometry were the followings: 35 a.u. sheath gas flow rate, 5000 V spray voltage, 275 °C capillary temperature, 31 V capillary voltage, 150 V tube lens voltage, and 34 V skimmer voltage.

Oxidation by synthetic porphyrin and the chemical Fenton system

Two reactions were carried out to test the stability of ERJ-500 molecule under oxidative conditions, based on the method as reported by Csepanyi et al. [2] recently, with minor modifications as follows: 50 μl of ERJ-500 dissolved in acetonitrile was used for synthetic porphyrin oxidation in 10 mM concentration. 400 μl of ERJ-500 in 2.5 mM concentration for the Fenton reaction. Samples were drawn at 1 h in the Fenton reactions prior to injecting them instantly to the HPLC and further investigation. Reaction mixtures for blank contained acetonitrile only without ERJ-500. The control mixtures contained no peroxide.

Isolated working heart preparation to assess cardiac parameters

To measure cardiac function (Aortic flow, Aortic pressure, Derivated aortic pressure, and Cardiac output), isolated working heart preparations were carried out based on a previously described method by Czompa et al. [3] on Sprague Dawley female rat hearts (n = 11 in the control group, n = 6 in the treated group). After completing the isolated working heart preparation procedure followed by 10 min washout period, and aorta flow, aortic pressure, derivated aortic pressure, and cardiac output were registered (Fig. 12). Cardiac output was calculated by the sum of aortic and coronary flow represented in the associated research article [1]. In the treated group, ERJ-500 was added to the KHB buffer by a dilution of a previously prepared stock solution leading to a 100 μM concentration of ERJ-500 in the inflow line. The molecule-containing KHB buffer was presented after the washout and baseline registration period for 5 min, followed by a 30 min ischemia and 90 min of reperfusion.

Specifications table

Subject area	Chemistry, Biology
More specific subject area	Organic chemistry, Pharmacology
Type of data	1H and13C NMR spectra, HPLC-MS chromatograms, column tables
How data was acquired	NMR Bruker DRX-400 spectrometer at 25 °C, HPLC-MSLTQ XL linear ion trap mass spectrometer coupled with Accela LC system (Thermo Fisher Scientific, Waltham, MA, USA)., “Isolated working heart system”.
Data format	Raw, filtered and analyzed
Experimental factors	Initial compounds were purchased, the intermediates and the end-product were synthesized as described in the original paper.
Experimental features	New compounds have been characterized by spectrometric methods and an ex vivo technique on rat hearts
Data source location	- Department of Pharmacology and Department of Bioanalytical Chemistry, Nagyerdei Krt 98, H-4032 Debrecen, Hungary- Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Debrecen, Egyetem tér 1, H-4032 Debrecen, Hungary
Data accessibility	Data are provided with this article.
Related research article	K. Szoke, A. Czompa, I. Lekli, P. Szabados-Furjesi, M. Herczeg, M. Csavas, A. Borbas, P. Herczegh, A. Tosaki, A new, vasoactive hybrid aspirin containing nitrogen monoxide-releasing molsidomine moiety, Eur. J. Pharm. Sci., 131, 2019, 159–166 [1].

Value of the data• The NMR spectra can be used for structure elucidation of similar synthetic compounds. • Dataset of stability tests could be used to pretest the degradation profiles of molecules planned to try in vivo circumstances. • The cardiac functions detailed here are important indicator related to the contractile activity of the myocardium (see Fig. 2).

Non-significant myocardial parameters in working heart preparation.

Aorta flow		Aortic pressure		Derivated aortic pressure		Cardiac output
Control	ERJ-500	Control	ERJ-500	Control	ERJ-500	Control	ERJ-500
22	46	87	100	740	1361	42	72
31	50	92,7	110	924	2159	51	73
25	32	110,9	104	1670	1666	39	58
48	38	99	106	1357	1938	69	57
42	42	102	102,6	1568	1783	62	66
34	42	105	109	1756	1786	54	66
40		106		1879		56
34		104		1873		52
46		99		1424		65
46		99		1558		65
46		113,5		1956		65