Scientific Reports
Label-free integrative pharmacology on-target of drugs at the2-adrenergic receptor
Ann M. Ferrie, Haiyan Sun, &Ye Fang*
Supplementary Table S1 / DrugBank ID, name, indications, and mechanism of action and targets of adrenergic receptor drugsSupplementary Table S2 / Source, catalogue number, and CAS number of adrenergic receptor drugs
Supplementary Table S3 / The AR drugs and their potency acting on 2AR
Supplementary Fig.S1 / Time domain clustering of all -drug DMR signals
Supplementary Fig.S2 / Heat map of all drugs using single time point DMR
Supplementary Fig.S3 / The -AR drug DMR in the DMSO treated cells
Supplementary Fig.S4 / The epinephrine DMR in the -AR drug treated cells
Supplementary Fig.S5 / The -AR drug DMR in the 1hr epinephrine activated cells
Supplementary Fig.S6 / The -AR drug DMR in the 5hr epinephrine pretreated cells
Supplementary Fig.S7 / The -AR drug DMR in the 1hr propranolol treated cells
Supplementary Fig.S8 / The propranolol DMR in the 5hr -AR drug-pretreated cells
Supplementary Fig.S9 / The DMR of forskolin in the absence and presence of a -AR drug
Supplementary Fig.S10 / The -AR drug DMR in the cholera toxin pretreated cells
Supplementary Fig.S11 / The -AR drug DMR in the U0126 pretreated cells
Supplementary Fig.S12 / The -AR drug DMR in the TBB pretreated cells
Supplementary Fig.S13 / Bar chart of DMR signals under 12 assay conditions
Supplementary Fig. S14 / The potency in DMR and cAMP of isoetharine
References
Supplementary Table S1.DrugBank ID, name, indications, and mechanism of action and targets of adrenergic receptor drugs examined [1].
Drug Bank ID / Name / Indication / Mechanism of Action / TargetsDB01193 / Acebutolol / For the management of hypertension and ventricular premature beats in adults. / Acebutolol is a selective 1-receptor antagonist. Activation of 1-receptors by epinephrine increases the heart rate and the blood pressure, and the heart consumes more oxygen. Acebutolol blocks these receptors which reverses the effects of epinephrine, lowering the heart rate and blood pressure. In addition, beta blockers prevent the release of renin, which is a hormone produced by the kidneys which leads to constriction of blood vessels. / 1-AR
DB00866 / Alprenolol / For the treatment of hypertension, angina, and arrhythmia. / Alprenolol non-selectively blocks 1 receptors mainly in the heart, inhibiting the effects of epinephrine and norepinephrine resulting in a decrease in heart rate and blood pressure. By binding 2 receptors in the juxtaglomerular apparatus, alprenolol inhibits the production of renin, thereby inhibiting angiotensin II and aldosterone production and therefore inhibits the vasoconstriction and water retention due to angiotensin II and aldosterone, respectively. / 1-AR
2-AR
DB00335 / Atenolol / For the management of hypertention and long-term management of patients with angina pectoris. / Atenolol competes with sympathomimetic neurotransmitters such as catecholamines for binding at 1 adrenergic receptors in the heart and vascular smooth muscle, inhibiting sympathetic stimulation. This results in a reduction in resting heart rate, cardiac output, systolic and diastolic blood pressure, and reflex orthostatic hypotension. Higher doses of atenolol also competitively block 2 adrenergic responses in the bronchial and vascular smooth muscles. / 1-AR
DB00195 / Betaxolol / For the management of hypertension. / Betaxolol selectively blocks catecholamine stimulation of 1-adrenergic receptors in the heart and vascular smooth muscle. This results in a reduction of heart rate, cardiac output, systolic and diastolic blood pressure, and possibly reflex orthostatic hypotension. Betaxolol can also competitively block 2-adrenergic responses in the bronchial and vascular smooth muscles, causing bronchospasm. / 1-AR
DB00612 / Bisoprolol / For the management of hypertension and prophylaxis treatment of angina pectoris and heart failure. / Bisoprolol selectively blocks catecholamine stimulation of 1-adrenergic receptors in the heart and vascular smooth muscle. This results in a reduction of heart rate, cardiac output, systolic and diastolic blood pressure, and possibly reflex orthostatic hypotension. Bisoprolol can also competitively block 2-adrenergic responses in the bronchial and vascular smooth muscles, causing bronchospasm. / 1-AR
2-AR
DB01136 / Carvedilol / For the treatment of mild or moderate (NYHA class II or III) heart failure of ischemic or cardiomyopathic origin. / Carvedilol is a racemic mixture in which nonselective beta-adrenoreceptor blocking activity is present in the S(-) enantiomer and -adrenergic blocking activity is present in both R(+) and S(-) enantiomers at equal potency. Carvedilol's -adrenergic receptor blocking ability decreases the heart rate, myocardial contractility, and myocardial oxygen demand. Carvedilol also decreases systemic vascular resistance via its -adrenergic receptor blocking properties. Carvedilol and its metabolite BM-910228 (a less potent beta blocker, but more potent antioxidant) have been shown to restore the inotropic responsiveness to Ca2+ in OH- free radical-treated myocardium. Carvedilol and its metabolites also prevent OH- radical-induced decrease in sarcoplasmic reticulum Ca2+-ATPase activity. Therefore, carvedilol and its metabolites may be beneficial in chronic heart failure by preventing free radical damage. / 1-AR
1-AR
2-AR
Supplementary Table S1 (Continued)
Drug Bank ID / Name / Indication / Mechanism of Action / TargetsDB01407 / Clenbuterol / Used as a bronchodilator in the treatment of asthma patients. / 2-AR
DB00841 / Dobutamine / For inotropic support in the short- term treatment of patients with cardiac decompensation due to depressed contractility resulting either from organic heart disease or from cardiac surgical procedures. / Dobutamine directly stimulates 1 receptors of the heart to increase myocardial contractility and stroke volume, resulting in increased cardiac output. / 1-AR
DB00988 / Dopamine / For the correction of hemodynamic imbalances present in the shock syndrome due to myocardial infarction, trauma, endotoxic septicemia, open-heart surgery, renal failure, and chronic cardiac decompensation as in congestive failure. / Dopamine is a precursor to norepinephrine in noradrenergic nerves and is also a neurotransmitter in certain areas of the central nervous system. Dopamine produces positive chronotropic and inotropic effects on the myocardium, resulting in increased heart rate and cardiac contractility. This is accomplished directly by exerting an agonist action on -adrenoceptors and indirectly by causing release of norepinephrine from storage sites in sympathetic nerve endings. / 1-AR
DB00668 / Epinephrine / Used to treat anaphylaxis and sepsis. / Epinephrine works via the stimulation of alpha and 1 adrenergic receptors, and a moderate activity at 2 adrenergic receptors. / 1A-AR
1-AR
1-AR
DB01288 / Fenoterol / For the treatment of asthma. / 2-receptor stimulation in the lung causes relaxation of bronchial smooth muscle, bronchodilation, and increased bronchial airflow. / 2-AR
DB00983 / Formoterol / For use as long-term maintenance treatment of asthma in patients 6 years of age and older with reversible obstructive airways disease, including patients with symptoms of nocturnal asthma, who are using optimal corticosteroid treatment and experiencing regular or frequent breakthrough symptoms requiring use of a short-acting bronchodilator. / The pharmacologic effects of 2-adrenoceptor agonist drugs, including formoterol, are at least in part attributable to stimulation of intracellular adenyl cyclase, the enzyme that catalyzes the conversion of adenosine triphosphate (ATP) to cyclic-3', 5'-adenosine monophosphate (cyclic AMP). Increased cyclic AMP levels cause relaxation of bronchial smooth muscle and inhibition of release of mediators of immediate hypersensitivity from cells, especially from mast cells. In vitro tests show that formoterol is an inhibitor of the release of mast cell mediators, such as histamine and leukotrienes, from the human lung. Formoterol also inhibits histamine-induced plasma albumin extravasation in anesthetized guinea pigs and inhibits allergen-induced eosinophil influx in dogs with airway hyper-responsiveness. The relevance of these in vitro and animal findings to humans is unknown. / 2-AR
DB00221 / Isoetharine / For the treatment of asthma, wheezing, and chronic asthmatic bronchitis. / The pharmacologic effects of isoetharine are attributable to stimulation through -adrenergic receptors of intracellular adenyl cyclase, the enzyme that catalyzes the conversion of adenosine triphosphate (ATP) to cyclic AMP. Increased cyclic AMP levels are associated with relaxation of bronchial smooth muscle and inhibition of release of mediators of immediate hypersensitivity from cells, especially from mast cells. / 1-AR
2-AR
Supplementary Table S1 (Continued)
Drug Bank ID / Name / Indication / Mechanism of Action / TargetsDB01064 / Isoproterenol / For the treatment of mild or transient episodes of heart block that do not require electric shock or pacemaker therapy also used in management of asthma and chronic bronchitis. / The pharmacologic effects of isoproterenol are at least in part attributable to stimulation through -adrenergic receptors of intracellular adenyl cyclase, the enzyme that catalyzes the conversion of adenosine triphosphate (ATP) to cyclic AMP. Increased cyclic AMP levels are associated with relaxation of bronchial smooth muscle and inhibition of release of mediators of immediate hypersensitivity from cells, especially from mast cells. / 1-AR
2-AR
DB00598 / Labetalol / For the management of hypertension. / Labetalol has two asymmetric centers and therefore, exists as a molecular complex of two diastereoisomeric pairs. Dilevalol, the R,R' stereoisomer, makes up 25% of racemic labetalol. Labetalol HCl combines both selective, competitive, 1-adrenergic blocking and nonselective, competitive, 1-adrenergic blocking activity in a single substance. In man, the ratios of - to - blockade have been estimated to be approximately 1:3 and 1:7 following oral and intravenous (IV) administration, respectively. 2-agonist activity has been demonstrated in animals with minimal beta-1-agonist (ISA) activity detected. In animals, at doses greater than those required for alpha- or beta- adrenergic blockade, a membrane stabilizing effect has been demonstrated. / 1A-AR
1B-AR
1-AR
2-AR
DB00968 / Methyldopa (methylnor-epinephrine) / For use in the treatment of hypertension. / Although the mechanism of action has yet to be conclusively demonstrated, the antihypertensive effect of methyldopa probably is due to its metabolism to alpha-methylnorepinephrine, which then lowers arterial pressure by stimulation of central inhibitory alpha-adrenergic receptors, false neurotransmission, and/or reduction of plasma renin activity. Methyldopa has been shown to cause a net reduction in the tissue concentration of serotonin, dopamine, norepinephrine, and epinephrine. / 2A-AR
DB00264 / Metoprolol / For the treatment of hypertension and angina pectoris. / Metoprolol competes with adrenergic neurotransmitters such as catecholamines for binding at 1-adrenergic receptors in the heart and vascular smooth muscle. 1-receptor blockade results in a decrease in heart rate, cardiac output, and blood pressure. / 1-AR
DB01203 / Nadolol / Used in cardiovascular disease to treat arrhythmias, angina pectoris, and hypertension. / Nadolol competes with adrenergic neurotransmitters such as catecholamines for binding at sympathetic receptor sites. Nadolol binds at 1-adrenergic receptors in the heart and vascular smooth muscle, inhibiting the effects of the catecholamines epinephrine and norepinephrine and decreasing heart rate, cardiac output, and systolic and diastolic blood pressure. It also blocks 2 adrenergic receptors located in bronchiole smooth muscle, causing vasoconstriction. By binding 2 receptors in the juxtaglomerular apparatus, nadolol inhibits the production of renin, thereby inhibiting angiotensin II and aldosterone production. Nadolol therefore inhibits the vasoconstriction and water retention due to angiotensin II and aldosterone, respectively. / 1-AR
2-AR
DB00368 / Norepinephrine / Mainly used to treat patients in vasodilatory shock states such as septic shock and neurogenic shock and has shown a survival benefit over dopamine. Also used as a vasopressor medication for patients with critical hypotension. / Norepinephrine functions as a peripheral vasoconstrictor (alpha-adrenergic action) and as an inotropic stimulator of the heart and dilator of coronary arteries (beta-adrenergic action). / 1A-AR
1B-AR
1D-AR
2A-AR
2B-AR
2C-AR
1-AR
2-AR
3-AR
Supplementary Table S1 (Continued)
Drug Bank ID / Name / Indication / Mechanism of Action / TargetsDB01580 / Oxprenolol / Used in the treatment of hypertension, angina pectoris, arrhythmias, and anxiety. / Oxprenolol competes with adrenergic neurotransmitters such as catecholamines for binding at sympathetic receptor sites. Oxprenolol binds at 1adrenergic receptors in the heart and vascular smooth muscle, inhibiting the effects of the catecholamines epinephrine and norepinephrine and decreasing heart rate, cardiac output, and systolic and diastolic blood pressure. It also blocks 2 adrenergic receptors located in bronchiole smooth muscle, causing vasoconstriction. By binding 2 receptors in the juxtaglomerular apparatus, oxprenolol inhibits the production of renin, thereby inhibiting angiotensin II and aldosterone production. Oxprenolol therefore inhibits the vasoconstriction and water retention due to angiotensin II and aldosterone, respectively. / 1-AR
2-AR
DB00388 / Phenylephrine / For the treatment of ophthalmic disorders (hyperaemia of conjunctiva, posterior synechiae, acute atopic), nasal congestion, hemorrhoids, hypotension, shock, hypotension during spinal anesthesia, paroxysmal supraventricular tachycardia. It is also used as an aid in the diagnosis of heart murmurs and for prolongation of spinal anesthesia. / Phenylephrine produces its ophthalmic and systemic actions by acting on 1 adrenergic receptors in the pupillary dilator muscle and the vascular smooth musle, resulting in contraction of the dilator muscle and contraction of the smooth muscle in the arterioles of the conjunctiva and peripheral vasoconstriction. Phenylephrine decreases nasal congestion by acting on 1 adrenergic receptors in the arterioles of the nasal mucosa to produce constriction. / 1A-AR
1B-AR
DB00960 / Pindolol / For the management of hypertension, edema, ventricular tachycardias, and atrial fibrillation. / Pindolol non-selectively blocks 1 adrenergic receptors mainly in the heart, inhibiting the effects of epinephrine and norepinephrine resulting in a decrease in heart rate and blood pressure. By binding 2 receptors in the juxtaglomerular apparatus, Pindolol inhibits the production of renin, thereby inhibiting angiotensin II and aldosterone production and therefore inhibits the vasoconstriction and water retention due to angiotensin II and aldosterone, respectively. / 1-AR
2-AR
3-AR
DB01297 / Practolol / Used in the emergency treatment of cardiac arrhythmias. / Practolol competes with adrenergic neurotransmitters such as catecholamines for binding at sympathetic receptor sites. Practolol binds at 1-adrenergic receptors in the heart and vascular smooth muscle, inhibiting the effects of the catecholamines epinephrine and norepinephrine and decreasing heart rate, cardiac output, and systolic and diastolic blood pressure. / 1-AR
DB01366 / Procaterol / For the treatment of asthma and chronic obstructive pulmonary disease (COPD). / 2-receptor stimulation in the lung causes relaxation of bronchial smooth muscle, bronchodilation, and increased bronchial airflow. / 2-AR
DB00571 / Propranolol / For the prophylaxis of migraine. / Propranolol competes with sympathomimetic neurotransmitters such as catecholamines for binding at 1 adrenergic receptors in the heart and vascular smooth muscle, inhibiting sympathetic stimulation. This results in a reduction in resting heart rate, cardiac output, systolic and diastolic blood pressure, and reflex orthostatic hypotension. Higher doses of atenolol also competitively block 2 adrenergic responses in the bronchial and vascular smooth muscles. / 1-AR
2-AR
3-AR
Supplementary Table S1 (Continued)
Drug Bank ID / Name / Indication / Mechanism of Action / TargetsDB00867 / Ritodrine / For the treatment and prophylaxis of premature labor. / Ritodrine is 2 adrenergic agonist. It binds to 2 adrenergic receptors on outer membrane of myometrial cell, activates adenyl cyclase to increase the level of cAMP which decreases intracellular calcium and leads to a decrease of uterine contractions. / 2-AR
DB01001 / Salbutamol / For relief and prevention of bronchospasm due to asthma, emphysema, and chronic bronchitis. / Salbutamol is a 2-adrenergic agonist and thus it stimulates 2-adrenergic receptors. Binding of albuterol to 2-receptors in the lungs results in relaxation of bronchial smooth muscles. It is believed that salbutamol increases cAMP production by activating adenylate cyclase, and the actions of salbutamol are mediated by cAMP. Increased intracellular cyclic AMP increases the activity of cAMP-dependent protein kinase A, which inhibits the phosphorylation of myosin and lowers intracellular calcium concentrations. A lowered intracellular calcium concentration leads to a smooth muscle relaxation. Increased intracellular cyclic AMP concentrations also cause an inhibition of the release of mediators from mast cells in the airways. / 2-AR
DB00938 / Salmeterol / For the treatment of asthma and chronic obstructive pulmonary disease (COPD). / Salmeterol's long, lipophilic side chain binds to exosites near 2-receptors in the lungs and on bronchiolar smooth muscle, allowing the active portion of the molecule to remain at the receptor site, continually binding and releasing. 2-receptor stimulation in the lung causes relaxation of bronchial smooth muscle, bronchodilation, and increased bronchial airflow. / 2-AR
DB00489 / Sotalol / For the maintenance of normal sinus rhythm [delay in time to recurrence of atrial fibrillation/atrial flutter (AFIB/AFL)] in patients with symptomatic AFIB/AFL who are currently in sinus rhythm. Also for the treatment of documented life-threatening ventricular arrhythmias. / Sotalol has both beta-adrenoreceptor blocking (Vaughan Williams Class I) and cardiac action potential duration prolongation (Vaughan Williams Class I) antiarrhythmic properties. Sotalol is a racemic mixture of d- and l-sotalol. Both isomers have similar Class I antiarrhythmic effects, while the l-isomer is responsible for virtually all of the beta-blocking activity. Sotalol inhibits response to adrenergic stimuli by competitively blocking β1-adrenergic receptors within the myocardium and β2-adrenergic receptors within bronchial and vascular smooth muscle. The electrophysiologic effects of sotalol may be due to its selective inhibition of the rapidly activating component of the potassium channel involved in the repolarization of cardiac cells. The class II electrophysiologic effects are caused by an increase in sinus cycle length (slowed heart rate), decreased AV nodal conduction, and increased AV nodal refractoriness, while the class III electrophysiological effects include prolongation of the atrial and ventricular monophasic action potentials, and effective refractory period prolongation of atrial muscle, ventricular muscle, and atrio-ventricular accessory pathways (where present) in both the anterograde and retrograde directions. / KCNH2
1-AR
2-AR
DB00871 / Terbutaline / For the prevention and reversal of bronchospasm in patients 12 years of age and older with asthma and reversible bronchospasm associated with bronchitis and emphysema. / The pharmacologic effects of terbutaline are at least in part attributable to stimulation through -adrenergic receptors of intracellular adenyl cyclase, the enzyme that catalyzes the conversion of adenosine triphosphate (ATP) to cyclic AMP. Increased cyclic AMP levels are associated with relaxation of bronchial smooth muscle and inhibition of release of mediators of immediate hypersensitivity from cells, especially from mast cells. / 2-AR
DB00373 / Timolol / In its oral form it is used to treat high blood pressure and prevent heart attacks, and occasionally to prevent migraine headaches. In its opthalmic form it is used to treat open-angle and occasionally secondary glaucoma. / Timolol competes with adrenergic neurotransmitters such as catecholamines for binding at 1-adrenergic receptors in the heart and vascular smooth muscle and 2-receptors in the bronchial and vascular smooth muscle. 1-receptor blockade results in a decrease in resting and exercise heart rate and cardiac output, a decrease in both systolic and diastolic blood pressure, and, possibly, a reduction in reflex orthostatic hypotension. 2-blockade results in an increase in peripheral vascular resistance. The exact mechanism whereby timolol reduces ocular pressure is still not known. The most likely action is by decreasing the secretion of aqueous humor. / 1-AR
2-AR
Supplementary Table S2.Source, catalogue number, and CAS number of adrenergic receptor drugs examined.