Patent Grant
7504421
The present invention relates to substituted 2-thio-3,5-dicyano-4-aryl-6-amino-pyridines, to a process for their preparation and to their use as active compounds for medicaments.
The present invention furthermore provides the use of adenosine-receptor-selective ligands for the prophylaxis and/or treatment of various disorders.
Adenosine, a nucleoside consisting of adenine and D-ribose, is an endogenous factor having cell-protective activity, in particular under cell-damaging conditions with limited oxygen and substrate supply, such as, for example, in the case of ischemia in various organs (for example heart and brain).
Adenosine is formed intracellularly as an intermediate during the degradation of adenosine 5′-monophosphate (AMP) and S-adenosylhomocysteine, but it can be released from the cell, in which case it acts as a hormone-like substance or neurotransmitter by binding to specific receptors.
Under normoxic conditions, the concentration of free adenosine in the extracellular space is very low. However, under ischemic or hypoxic conditions, the extracellular concentration of adenosine in the affected organs is increased dramatically. Thus, it is known, for example, that adenosine inhibits platelet degradation and increases the blood supply to the coronary vessels of the heart. Furthermore, it acts on the heart rate, on the release of neurotransmitters and on lymphocyte differentiation.
The aim of these actions of adenosine is to increase the oxygen supply of the affected organs and/or to reduce the metabolism of these organs in order to adjust the metabolism of the organ to the blood supply of the organ under ischemic or hypoxic conditions.
The action of adenosine is mediated via specific receptors. To date, subtypes A1, A2a, A2b and A3 are known. The actions of these adenosine receptors are mediated intracellularly by the messenger cAMP. In the case of the binding of adenosine to the A2a or A2b receptors, the intracellular cAMP is increased via activation of the membrane-bond adenylate cyclase, whereas binding of adenosine to A1 or A3 receptors results in a decrease of the intracellular cAMP concentration via inhibition of adenylate cyclase.
According to the invention, “Adenosine-receptor-selective ligands” are substances which bind selectively to one or more subtypes of the adenosine receptors, thus either mimicking the action of adenosine (adenosine agonists) or blocking its action (adenosine antagonists).
According to their receptor selectivity, adenosine-receptor-selective ligands can be divided into different categories, for example ligands which bind selectively to the A1 or A2 receptors of adenosine and in the case of the latter also, for example, those which bind selectively to the A2a or the A2b receptors of adenosine. Also possible are adenosine receptor ligands which bind selectively to a plurality of subtypes of the adenosine receptors, for example ligands which bind selectively to the A1 and the A2, but not to the A3 receptors of adenosine.
The abovementioned receptor selectivity can be determined by the effect of the substances on cell lines which, after stable transfection with the corresponding cDNA, express the receptor subtypes in question (see the publication M. E. Olah, H. Ren, J. Ostrowski, K. A. Jacobson, G. L. Stiles, “Cloning, expression, and characterization of the unique bovine A1 adenosine receptor. Studies on the ligand binding site by site-directed mutagenesis.” in J. Biol. Chem. 267 (1992) pages 10764-10770, the disclosure of which is hereby fully incorporated by way of reference).
The effect of the substances on such cell lines can be monitored by biochemical measurement of the intracellular messenger cAMP (see the publication K. N. Klotz, J. Hessling, J. Hegler, C. Owman, B. Kull, B. B. Fredholm, M. J. Lohse, “Comparative pharmacology of human adenosine receptor subtypes—characterization of stably transfected receptors in CHO cells” in Naunyn Schmiedebergs Arch. Pharmacol. 357 (1998) pages 1-9, the disclosure of which is hereby fully incorporated by way of reference).
The “adenosine-receptor-specific” ligands known from the prior art are mainly derivatives based on natural adenosine (S.-A. Poulsen and R. J. Quinn, “Adenosine receptors: new opportunities for future drugs” in Bioorganic and Medicinal Chemistry 6 (1998) pages 619-641). However, most of the adenosine ligands known from the prior art have the disadvantage that their action is not really receptor-specific, that their activity is less than that of natural adenosine or that they have only very weak activity after oral administration. Thus, because of the disadvantages mentioned above, they are mainly only used for experimental purposes.
It is now an object of the following invention to find or provide compounds which have a wide therapeutic range and can serve as active compounds for the prophylaxis and/or treatment of various diseases.
In particular, it is an object of the present invention to find or provide substances which preferably act as adenosine-receptor-selective ligands and are suitable for the prophylaxis and/or treatment of various disorders, in particular disorders of the cardiovascular system (cardiovascular disorders) or inflammatory disorders, but additionally also disorders of the urogenital system, the respiratory tract, the central nervous system, the diabetes (in particular diabetes mellitus) and cancer.
It is a further object of the present invention to find or provide adenosine-receptor-selective ligands having a high specificity of action for the abovementioned purposes.
Accordingly, the present invention relates to compounds of the general formula (I)
in which:
Some of the abovementioned substances which can be used in accordance with the present invention for the prophylaxis and/or treatment of disorders are novel, and some are also known from the literature (see Dyachenko et al., Russian Journal of Chemistry, Vol. 33, No. 7, 1997, pages 1014-1017 and Vol. 34, No. 4, 1998, pages 557-563; Dyachenko et al., Chemistry of Heterocyclic Compounds, Vol. 34, 1998, pages 188-194; Elnagdi et al., Zeitschrift für Naturforschung, Vol. 47b, 1992, pages 572-578; Riguera et al., Eur. J. Med. Chem. 33, 1998, pages 887-897; J. Vaquero, Thesis, University of Alcala de Henares, Madrid, Spain, 1981). However, in the literature, a therapeutic use of the known compounds has hitherto not been described. The first time this has happened is in the context of the present invention.
Accordingly, the present invention also provides the use of the abovementioned compounds of the general formula (I), including the compounds excluded above, for the prophylaxis and/or treatment of disorders.
Depending on the substitution pattern, the compounds of the formula (I) can exist in stereoisomeric forms which are either like image and mirror image (enantiomers) or not like image and mirror image (diastereomers). The invention relates both to the enantiomers or diastereomers and to their respective mixtures. The racemic forms, like the diastereomers, can be separated in a known manner into the stereoisomerically uniform components. Likewise, the present invention also relates to the other tautomers of the compounds of the formula (I) and their salts.
Physiologically acceptable salts of the compounds of the formula (I) can be salts of the substances according to the invention with mineral acids, carboxylic acids or sulfonic acids. Particular preference is given, for example, to salts with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, trifluoroacetic acid, acetic acid, propionic acid, lactic acid, tartaric acid, citric acid, fumaric acid, maleic acid or benzoic acid.
Salts which may be mentioned include salts with customary bases, such as, for example, alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example calcium or magnesium salts) or ammonium salts, derived from ammonia or organic amines such as, for example, diethylamine, triethylamine, ethyldiisopropylamine, procaine, dibenzylamine, N-methylmorpholine, dihydroabietylamine, 1-ephenamine or methylpiperidine.
Definitions in the Context of the Present Invention:
Compounds which are preferred in the context of the invention are compounds of the general formula (I),
in which:
except for the following compounds of the general formula (I), in which the radicals R1, R2, R3 and R4 are as defined below:
Particularly preferred compounds are the compounds of the general formula (I)
in which:
except for the following compounds of the general formula (I), in which the radicals R1, R2, R3 and R4 are as defined below:
Particular preference according to the invention is given to compounds of the general formula (I)
in which:
or
except for the following compounds of the general formula (I), in which the radicals R1, R2, R3 and R4 are as defined below:
Very particular preference according to the invention is given to compounds of the general formula (I)
in which:
or
except for the following compounds of the general formula (I), in which the radicals R1, R2, R3 and R4 are as defined below:
The present invention also provides a process for preparing the compounds of the general formula (I).
According to a first variant of the process according to the invention, the compounds of the general formula (I) are prepared by
reacting compounds of the general formula (II)
in which the radicals R1, R2 and R3 are as defined above
with compounds of the general formula (III)
R4—X (III),
in which R4 is as defined above
and
X represents a nucleofugic group (preferably halogen, in particular chlorine, bromine or iodine, or mesylate, tosylate, triflate or 1-imidazolyl),
in inert solvents, if appropriate in the presence of a base.
The process described above can be illustrated in an exemplary manner by the equation below:
If the radical R4 in the general formula (I) has the meaning of
alkyl, substituted by the radicals —NR6—C(O)—R8, —NR6—C(O)—NR6R7, —NR6—SO2—R8,
where the radicals R6, R7 and R8 are as defined above,
it is alternatively, according to a second variant of the process according to the invention, also possible to prepare the compounds of the general formula (I) by initially reacting the compounds of the general formula (II) with 2-bromoethylamine to give compounds of the general formula (IV)
which are then reacted with compounds of the general formula
R9—Y (V),
in which
The second variant, described above, of the process according to the invention can be illustrated in an exemplary manner by the following equation:
The nucleofugic group X, which is sometimes also referred to as leaving group, can be introduced into the reaction separately or else be generated in situ by customary methods, for example by the “Mitsunobu reaction”.
Suitable solvents for the process according to the invention are all organic solvents which are inert under the reaction conditions. These include alcohols such as methanol, ethanol and isopropanol, ketones such as acetone and methyl ethyl ketone, acyclic and cyclic ethers such as diethyl ether and tetrahydrofuran, esters such as ethyl acetate or butyl acetate, hydrocarbons such as benzene, xylene, toluene, hexane or cyclohexane, dimethylformamide, acetonitrile, pyridine, dimethyl sulfoxide (DMSO), chlorinated hydrocarbons such as dichloromethane, chlorobenzene or dichloroethane or hexamethylphosphoric triamide. Water is also suitable for use as solvent. Particular preference is given to dimethylformamide. It is also possible to use mixtures of the solvents mentioned above.
Suitable bases are the customary inorganic or organic bases. These preferably include alkali metal hydroxides, such as, for example, sodium hydroxide or potassium hydroxide, or alkali metal carbonates, such as sodium carbonate or potassium carbonate or sodium bicarbonate or potassium bicarbonate, or sodium methoxide or potassium methoxide or sodium ethoxide or potassium ethoxide or potassium tert-butoxide, or else amides, such as sodium amide, lithium bis-(trimethylsilyl)amide or lithium diisopropylamide, or organometallic compounds, such as butyllithium or phenyllithium, or else amines, such as triethylamine and pyridine. Preference is given to the alkali metal carbonates and alkali metal bicarbonates.
Here, the base can be employed in an amount of from 1 to 10 mol, preferably from 1 to 5 mol, in particular from 1 to 4 mol, based on 1 mol of the compounds of the general formula (II) or (IV).
The reaction generally takes place in a temperature range of from −78° C. to reflux temperature. preferably in the range from −78° C. to +40° C., in particular at room temperature.
The reaction can be carried out at atmospheric, elevated or reduced pressure (for example in the range from 0.5 to 5 bar). In general, the reaction is carried out at atmospheric pressure.
The person skilled in the art is familiar with numerous modifications of the conditions mentioned above which are within the knowledge of the average expert and within the scope of the present invention.
The compounds of the general formulae (II) are likewise known per se to the person skilled in the art or can be prepared by customary methods known from the literature. Reference may be made in particular to the following publications, the respective content of which is expressly incorporated herein by way of reference:
It is also possible to prepare the compounds of the general formula (II) from compounds of the general formula (VI) by reaction with an alkali metal sulfide. This preparation method can be illustrated in an exemplary manner by the equation below:
The alkali metal sulfide used is preferably sodium sulfide in an amount of from 1 to 10 mol, preferably from 1 to 5 mol, in particular from 1 to 4 mol, based on 1 mol of the compounds of the general formula (VI).
Suitable solvents are all organic solvents which are inert under the reaction conditions. These include N,N-dimethylformamide, N-methylpyrrolidinone, hexamethylphosphoric triamide, pyridine and acetonitrile. Particularly preference is given to N,N-dimethylformamide. It is also possible to use mixtures of the solvents mentioned above.
The reaction is generally carried out in a temperature range of from +20° C. to reflux temperature, preferably in the range from +20° C. to +120° C., in particular at from +60° C. to +100° C.
The reaction can be carried out at atmospheric, elevated or reduced pressure (for example in the range from 0.5 to 5 bar). In general, the reaction is carried out at atmospheric pressure.
The person skilled in the art is familiar with numerous modifications of the conditions mentioned above which are within the knowledge of the average expert and within the scope of the present invention.
The compounds of the general formulae (VI) are likewise known per se to the person skilled in the art or can be prepared by customary methods known from the literature. Reference may be made in particular to the publication Kambe et al., Synthesis, 531 (1981) the content of which is expressly incorporated herein by way of reference.
The compounds of the general formulae (III) or (V) are either commercially available or known per se to the person skilled in the art or can be prepared by customary methods.
Surprisingly, the compounds of the general formula (I) have an unforeseeable useful pharmacological activity spectrum and are therefore suitable in particular for the prophylaxis and/or treatment of disorders.
This is because it has now been found, unexpectedly, that the substances of the formula (I) above are suitable for the prophylaxis and/or treatment of a large number of disorders, i.e. in particular, for example, disorders of the cardiovascular system (cardiovascular disorders); urogenital disorders; respiratory disorders; inflammatory and neuroinflammatory disorders; diabetes, in particular diabetes mellitus; cancer; and finally also neurodegenerative disorders, such as, for example, Parkinson's disease, and also pain.
In the context of the present invention, cardiovascular disorders are to be understood as meaning, in particular, for example the following disorders: coronary heart disease; hypertension (high blood pressure); restinosis such as, for example, restinosis after balloon dilatation of peripheral blood vessels; arteriosclerosis; tachycardia; arrhythmias; peripheral and cardiovascular disorders; stable and unstable angina pectoris; and atrial fibrillation.
The compounds of the general formula (I) are furthermore also suitable for reducing the myocardial area effected by an infarct.
The compounds of the general formula (I) are furthermore suitable for the treatment and prophylaxis of thromboembolic disorders and ischemias such as myocardial infarction, stroke and transitory ischemic attacks.
A further area of indication for which the compounds of the general formula (I) are suitable is the prophylaxis and/or therapy of urogenital disorders, such as, for example, irritable bladder, erectile dysfunction and female sexual dysfunction, and additionally also the prophylaxis and/or treatment of inflammatory disorders, such as, for example, asthma and inflammable dermatoses, of neuroinflammatory disorders of the central nervous system, such as, for example, conditions after cerebral infarction, Alzheimer's disease, furthermore also neurodegenerative disorders such as Parkinson's disease, and also pain.
A further area of indication is respiratory disorders such as, for example, asthma, chronic bronchitis, pulmonary emphysema, bronchiectases, cystic fibrosis (mucoviscidosis) and pulmonary hypertension.
The compounds of the general formula (I) are furthermore also suitable for the prophylaxis and/or therapy of hepatic fibrosis and cirrhosis of the liver.
Finally, the compounds of the general formula (I) are also suitable for the prophylaxis and/or therapy of diabetes, in particular diabetes mellitus.
Accordingly, the present invention also relates to the use of the substances of the general formula (I) for preparing medicaments and pharmaceutical compositions for the prophylaxis and/or treatment of the clinical features mentioned above.
The present invention furthermore relates to a method for the prophylaxis and/or the treatment of the clinical pictures mentioned above using substances of the general formula (I).
The pharmaceutical activity of the abovementioned compounds of the general formula (I) can be explained by their action as selective ligands on individual subtypes or a plurality of subtypes of the adenosine receptors, in particular as selective ligands on adenosine A1, adenosine A2a and/or adenosine A2b receptors, preferably as selective ligands on adenosine A1 and/or adenosine A2b receptors.
In the context of the present invention, “selective” are adenosine receptor ligands where, firstly, a clear effect on one or more adenosine receptor subtypes and, secondly, no or a considerably weaker effect on one or more other adenosine receptor subtypes can be observed, where, with respect to the test methods for the selectivity of action, reference is made to the test methods described in section A. II.
Compared to adenosine receptor ligands of the prior art, the substances of the general formula (I) are much more selective. Thus, for example, compounds of the general formula (I) in which R4 represents (C1-C4)-alkyl which is substituted by a group of the formula —C(O)NR6R7, where R6 and R7 are identical or different and are hydrogen or optionally substituted (C1-C3)-alkyl, are generally selective on adenosine A2b receptors.
On the other hand, compounds of the general formula (I) in which R4 represents (C1-C4)-alkyl substituted by one or more hydroxyl groups generally act selectively on adenosine A1 receptors.
Compounds of the general formula (I) in which R4 represents (C1-C4)-alkyl substituted by imidazolyl or optionally substituted benzyl, in turn, generally act selectively on adenosine A1 and adenosine A2b receptors.
This receptor selectivity can be determined by biochemical measurement of the intracellular messenger cAMP in cells which specifically only express one subtype of the adenosine receptors. In the case of agonists, an increase in the intracellular cAMP concentration is observed; in the case of antagonists, a decrease in the intracellular cAMP concentration after prior stimulation with adenosine or adenosine-like substances is observed (see the publications B. Kull, G. Arslan, C. Nilsson, C. Owman, A. Lorenzen, U. Schwabe, B. B. Fredholm, “Differences in the order of potency for agonists but not antagonists at human and rat adenosine A2A receptors”, Biochem. Pharmacol., 57 (1999) pages 65-75; and S. P. Alexander, J. Cooper, J. Shine, S. J. Hill, “Characterization of the human brain putative A2B adenosine receptor expressed in Chinese hamster ovary (CHO.A2B4) cells”, Br. J. Pharmacol., 119 (1996) pages 1286-90, the respective disclosure of which is hereby expressly incorporated by way of reference).
Accordingly, the present invention also provides the use of selective adenosine receptor ligands, in particular of selective adenosine A1, adenosine A2a and/or adenosine A2b receptor ligands, for preparing medicaments and pharmaceutical compositions for the prophylaxis and/or treatment of disorders, in particular, for example, disorders of the cardiovascular system (cardiovascular disorders); urogenital disorders; inflammatory and neuroinflammatory disorders; neurodegenerative disorders; respiratory disorders; hepatic fibrosis, cirrhosis of the liver; cancer; and finally diabetes, in particular diabetes mellitus, where, with respect to the individual areas of indication, reference is also made to what has been said above.
Thus, compounds of the general formula (I) which bind selectively to adenosine A1 receptors are preferably suitable for myocardial protection and for the prophylaxis and/or treatment of tachycardias, atrial arrhythmias, cardiac insufficiency, of acute kidney failure, diabetes and of pain. Compounds of the general formula (I) which bind selectively to adenosine A2a receptors, on the other hand, are preferably suitable for the prophylaxis and/or treatment of thromboembolic disorders, of neurodengenerative disorders such as Parkinson's disease and also for wound healing. Compounds of the general formula (I) which bind selectively to adenosine A2b receptors, in turn, are preferably suitable for the prophylaxis and/or therapy of hepatic fibrosis, of myocardial infarction, of neuroinflammatory disorders, Alzheimer's disease, of urogenital incontinence and also of respiratory disorders such as, for example, asthma and chronic bronchitis.
The present invention also provides medicaments and pharmaceutical compositions comprising at least one selective adenosine and/or adenosine A2b receptor ligand, preferably at least one compound of the general formula (I), together with one or more pharmacologically acceptable excipients or carriers, and their use for the purposes mentioned above.
Suitable for administering the compounds of the general formula (I) are all customary administration forms, i.e. oral, parenteral, inhalative, nasal, sublingual, rectal or external, such as, for example, transdermal, with particular preference oral or parenteral. In the case of parenteral administration, particular mention may be made of intravenous, intramuscular and subcutaneous administration, for example as a subcutaneous depot. Very particular preference is given to oral administration.
Here, the active compounds can be administered on their own or in the form of preparations. Suitable preparations for oral administration are inter alia tablets, capsules, pellets, sugar-coated tablets, pills, granules, solid and liquid aerosols, syrups, emulsions, suspensions and solutions. Here, the active compound has to be present in such a quantity that a therapeutic effect is obtained. In general, the active compound can be present in a concentration of from 0.1 to 100% by weight, in particular from 0.5 to 90% by weight, preferably from 5 to 80% by weight. In particular, the concentration of the active compound should be 0.5-90% by weight, i.e. the active compound should be present in quantitites sufficient to achieve the dosage range mentioned.
To this end, the active compounds can be converted in a manner known per se into the customary preparations. This is achieved using inert nontoxic pharmaceutically suitable carriers, excipients, solvents, vehicles, emulsifiers and/or dispersants.
Excipients which may be mentioned are, for example: water, nontoxic organic solvents such as, for example, paraffins, vegetable oils (for example sesame oil), alcohols (for example ethanol, glycerol), glycols (for example polyethylene glycol), solid carriers, such as natural or synthetic ground minerals (for example talc or silicates), sugars (for example lactose), emulsifiers, dispersants (for example polyvinylpyrrolidone) and glidants (for example magnesium sulfate).
In the case of oral administration, tablets may, of course, also contain additives such as sodium citrate, together with adjuvants such as starch, gelatin and the like. Aqueous preparations for oral administration may furthermore be admixed with flavor enhancers or colorants.
In general, it has been found to be advantageous to administer, in the case of parenteral administration, quantities of from about 0.1 to about 10 000 μg/kg, preferably from about 1 to about 1 000 μg/kg, in particular from about 1 μg/kg to about 100 μg/kg, of bodyweight, to obtain effective results. In the case of oral administration, the quantity is from about 0.1 to about 10 mg/kg, preferably from about 0.5 to about 5 mg/kg, in particular from about 1 to about 4 mg/kg, of bodyweight.
In spite of this, it may still be required, depending on bodyweight, administration route, individual response to the active compound, the type of preparation and the time or interval at which administration takes place, to deviate from the quantities mentioned.
The present invention is illustrated by the examples below; however, these examples are only intended to facilitate a better understanding of the invention and not to restrict the invention in any way.
I. Demonstration of the Cardiovascular Action
Langendorff Heart of the Rat:
After opening the thoracic cage of anesthetized rats, the heart is removed quickly and introduced into a conventional Langendorff apparatus. The coronary arteries are perfused at a constant volume (10 ml/min), and the resulting perfusion pressure is recorded via an appropriate pressure sensor. In this arrangement, a decrease of the perfusion pressure corresponds to a relaxation of the coronary arteries. At the same time, the pressure which is developed by the heart during each contraction is measured via a balloon introduced into the left ventricle and a further pressure sensor. The frequency of the isolated beating heart is calculated from the number of contractions per time unit.
In this test arrangement, the following values were obtained for the coronary perfusion pressure (the stated percentage refers to the reduction of the coronary perfusion pressure in percent at the respective concentration):
At the stated concentrations, the substances tested had no effect on the pressure developed during the contraction in the left ventricle and no effect on the heart rate. Thus, it was demonstrated that the substances act selectively only on coronary perfusion.
II. Demonstration of the Receptor Selectivity (Adenosine A1, A2a. A2b and A3 Receptor Selectivity)
Cells of the permanent line CHO (Chinese Hamster Ovary) were stably transfected with cDNA for the adenosine receptor subtypes A1, A2a, A2b and A3. Binding of the substances to the A2a or A2b receptor subtypes was determined by measuring the intracellular cAMP concentration in these cells using a conventional radioimmunological assay (cAMP-RIA, IBL GmbH, Hamburg, Germany).
In the case of the action of the substances as agonists, binding of the substances is expressed in an increase of the intracellular cAMP concentration. In these experiments, the adenosine analogue NECA (5-N-ethylcarboxamido-adenosine), which binds unselectively, but with high affinity, to all adenosine receptor subtypes and has agonistic action (Klotz, K. N., Hessling, J., Hegler, J., Owman, C., Kull, B., Fredholm, B. B., Lohse, M. J., Comparative pharmacology of human adenosine receptor subtypes—characterization of stably transfected receptors in CHO cells, Naunyn Schmiedebergs Arch Pharmacol, 357 (1998), 1-9) was used as reference compound.
The adenosine receptors A1 and A3 are coupled to a Gi protein, i.e. stimulation of these receptors results in an inhibition of adenylate cyclase and thus a reduction of the intracellular cAMP level. To identify A1/A3 receptor agonists, the adenylate cyclase is stimulated using forskolin. However, additional stimulation of the A1/A3 receptors inhibits adenylate cyclase, so that it is possible to detect A1/A3 receptor agonists by a comparatively low concentration of cAMP in the cell.
To demonstrate an antagonistic effect on adenosine receptors, the recombinant cells transfected with the corresponding receptor were prestimulated with NECA, and the effect of the substances on a reduction of the intracellular cAMP concentration by this prestimulation was examined. In these experiments, XAC (xanthine amine congener), which binds unselectively, but with high affinity, to all adenosine receptor subtypes and has antagonistic action (Müller, C. E., Stein, B., Adenosine receptor antagonists: structures and potential therapeutic applications, Current Pharmaceutical Design, 2 (1996), 501-530) was used as reference compound.
In the experiments below, the intracellular cAMP concentration in CHO cells which had been transfected with cDNA for the A2b receptor was determined. What is stated is the cAMP concentration in percent in all cells in a well of a microtiter plate, based on the control value obtained without any substances acting on the cells:
In these experiments, it was possible to block the action of all substances by the antagonist XAC, which is unselective, but highly specific for adenosine receptors.
In the experiments below, the intracellular cAMP concentration in CHO cells which had been transfected with cDNA for the A2a receptor was determined. What is stated is the cAMP concentration in percent in all cells in a well of a microtiter plate, based on the control value obtained without any substances acting on the cells:
In these experiments, it was possible to block the action of all substances by the antagonist XAC, which is unselective, but highly specific for adenosine receptors.
In the following experiments, the intracellular cAMP concentration in CHO cells transfected with the cDNA for the A1 receptor was determined. What is stated is the cAMP concentration in percent in all cells of a well of a microtiter plate, based on the control value obtained without the action of any substance but after prestimulation with 1 μM of forskolin for 15 min (in these measurements, the cAMP concentration without prestimulation with forskolin is 18%):
Thus, the compound of example A 1 has a clear agonistic effect on cells which express the adenosine receptor A2b and virtually no effect on cells with the A2a receptor. In contrast, the compounds from example A 43 and A 104 have a clear agonistic effect on cells with the A 1 receptor, virtually no effect on cells with A2a receptors and a considerably weaker effect on cells with the A2b receptor and are therefore selective adenosine A1 receptor agonists. The compound of example A 379, on the other hand, has a clear agonistic effect on cells with the A2b receptor, virtually no effect on cells with A2a receptors and a comparably weaker effect on cells with the A1 receptor and is thus a selective adenosine A2b receptor agonist.
At room temperature (RT), 53.6 mg (0.2 mmol) of 2-amino-4-(4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile and 45.6 mg (0.3 mmol) of N-methylbromo-acetamide are stirred in 0.5 ml of dimethylformamide (DMF) together with 33.6 mg (0.4 mmol) of NaHCO3 for 4 hours. Thin-layer chromatography (TLC) (CH2Cl2/CH3OH 10:1) shows complete conversion. The entire mixture is diluted with water and ethyl acetate (EA) and the EA phase is dried with MgSO4 and concentrated under reduced pressure. The residue crystallizes from methanol.
Yield: 45 mg (66.3% of theory), white crystals
Mass spectrum: molecular mass calculated: 339, found [M+H]+=340.3
At RT, 53.6 mg (0.2 mmol) of 2-amino-4-(4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile and 58.2 mg (0.3 mmol) of N,N-diethylbromoacetamide are stirred in 0.5 ml of DMF together with 33.6 mg (0.4 mmol) of NaHCO3 for 4 hours. TLC (CH2Cl2/CH3OH 10:1) shows complete conversion. The entire mixture is diluted with water and ethyl acetate and the EA phase is dried with MgSO4 and concentrated under reduced pressure. The residue crystallizes from methanol.
Yield: 50 mg (65.5% of theory), white crystals
Mass spectrum: molecular mass calculated: 381, found [M+H]+=382
At RT 0.76 g (2 mmol) of 2-amino-4-(4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile and 0.5 g (3 mmol) of N-ethylbromoacetamide are stirred in 5 ml of DMF together with 0.34 g (4 mmol) of NaHCO3 for 4 hours. After dilution with water, the mixture is extracted with ethyl acetate and the ethyl acetate phase is dried with MgSO4 and concentrated under reduced pressure. The solid residue obtained after concentration is stirred with methanol. The crystals are filtered off with suction and dried under reduced pressure.
Yield: 0.49 g (69.5% of theory), crystals
Mass spectrum: molecular mass calculated: 353, found [M+H]+=354.2
268 mg (1 mmol) of 2-amino-4-(4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile, 105 mg (1 mmol) of 2-bromoethylamine hydrobromide and 168 mg (2 mmol) of NaHCO3 are stirred in 1 ml of DMF for 1 hour. The entire mixture is diluted with a few milliliters of 1N HCl. The crystals are filtered off with suction and dried under reduced pressure.
Yield: 200 mg (64.2% of theory), yellow crystals
Mass spectrum: molecular mass calculated: 311, found [M+H]+=312
At RT 60 mg (0.2 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile and 30 mg (0.3 mmol) of N-acetylimidazole are stirred in 0.5 ml of DMF for 1 hour. Water is slowly added dropwise, the mixture becomes slightly turbid and the crude product crystallizes out. The product is filtered off with suction, washed with water and dried under reduced pressure. This gives 53 mg of yellow crystals. The crystals are dissolved in 1 ml of a 1:1 mixture of CH2Cl2/CH3OH, and a few drops of concentrated ammonia are added (removal of diacetylated byproduct). The mixture is stirred at RT for 5 hours. The product crystallizes out when the reaction solution is concentrated. The product is filtered off with suction and washed with methanol.
Yield: 37 mg (52.3% of theory), almost white crystals
Mass spectrum: molecular mass calculated: 353, found [M+H]+=354
Under argon, 31.1 mg (0.1 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended at RT in 1 to 2 ml of dichloromethane, and the mixture is cooled to from −20 to −25° C. At this temperature, 30.3 mg (0.3 mmol) of triethylamine and 28,3 mg (0.3 mmol) of methyl chloroformate are added. The mixture is stirred at −20 C for 30 minutes and then allowed to warm to 0° C. over a period of 1 hour. The mixture is concentrated under reduced pressure, 4 ml of a 2 molar NH3 solution in methanol are added and the mixture is stirred at RT for 1 hour. The mixture is then concentrated, dissolved in 600 μl of DMSO and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 600 μl of DMSO solution
Yield: 21.7 mg (58.7% of theory) of product
Mass spectrum: molecular mass calculated: 369, found [M+H]+=370.1
Under argon, 31.1 mg (0.1 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended at RT in 1 to 2 ml of dichloromethane, and the mixture is cooled to from −20 to −25° C. At this temperature, 30.3 mg (0.3 mmol) of triethylamine and 32.6 mg (0.3 mmol) of ethyl chloroformate are added. The mixture is stirred at −20° C. for 30 minutes and then allowed to warm to 0° C. over a period of 1 hour. The mixture is concentrated under reduced pressure, 4 ml of a 2 molar NH3 solution in methanol are added and the mixture is stirred at RT for 1 hour. The mixture is then concentrated, dissolved in 600 μl of DMSO and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 600 μl of DMSO solution
Yield: 20.5 mg (53.5% of theory) of product
Mass spectrum: molecular mass calculated: 383, found [M+H]+=384.2
Under argon, 31.1 mg (0.1 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended at RT in 1 to 2 ml of dichloromethane, and the mixture is cooled to from −20 to −25° C. At this temperature, 10.1 mg (0.1 mmol) of triethylamine and 9.4 mg (0.1 mmol) of methyl chloroformate are added. The mixture is stirred at −20° C. for 30 minutes and then allowed to warm to 0° C. over a period of 1 hour. The mixture is then concentrated dissolved in 600 μl of DMSO and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 600 μl of DMSO solution
Yield: 11.2 mg (26.2% of theory) of product
Mass spectrum: molecular mass calculated: 427, found [M+H]+=428.2
Under argon 31.1 mg (0.1 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended at RT in 1 to 2 ml of dichloromethane, and the mixture is cooled to from −20 to −25° C. At this temperature, 10.1 mg (0.1 mmol) of triethylamine and 10.9 mg (0.1 mmol) of ethyl chloroformate are added. The mixture is stirred at −20° C. for 30 minutes and then allowed to warm to 0° C. over a period of 1 hour. The mixture is then concentrated, dissolved in 600 μl of DMSO and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 600 μl of DMSO solution
Yield: 15.2 mg (33.4% of theory) of product
Mass spectrum: molecular mass calculated: 455, found [M+H]+=456.2
31.1 mg (0.1 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended in 0.91 ml of 1N HCl, and 8.1 mg (0.1 mmol) of potassium cyanate are added. After the addition of a few drops of methanol, the mixture is stirred at 50° C. for a total of 10 hours. The crystals are filtered off with suction and washed with water and ether.
Yield: 16 mg (45.1% of theory) of product
Mass spectrum: molecular mass calculated: 354, found [M+H]+=355.1
62.2 mg (0.2 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended in 0.4 ml of DMF, and 11.4 mg (0.2 mmol) of methyl isocyanate are added at room temperature. The mixture is stirred overnight, filtered and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 45.9 mg (62.3% of theory) of product
Mass spectrum: molecular mass calculated: 368, found [M+H]+=369.2
62.2 mg (0.2 mmol) of 2-amino-6-[(2-aminoethyl)sulfanyl]-4-(4-hydroxyphenyl)-3,5-pyridinedicarbonitrile are suspended in 0.4 ml of DMF, and 14.2 mg (0.2 mmol) of ethyl isocyanate are added at room temperature. The mixture is stirred overnight, filtered and purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 37.6 mg (49.2% of theory) of product
Mass spectrum: molecular mass calculated: 382, found [M+H]+=383.2
337.2 mg (1 mmol) of 2-amino-4-(3,5-dichloro-4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile and 207 mg (1.5 mmol) of bromoacetamide are dissolved in 4 ml of DMF, 336 mg (4 mmol) of NaHCO3 are added and the mixture is stirred at RT for 8 hours. The mixture is diluted with water and washed with ethyl acetate. The aqueous phase is acidified with 1N HCl and the resulting crystals are filtered off with suction and dried.
Yield: 180 mg (45.7% of theory) of product
Mass spectrum: molecular mass calculated: 393, found [M+H]+=394.1
84 mg (0.163 mmol) of 2-amino-4-{4-[(4-methylpiperazino)sulfonyl]phenyl}-6-sulfanyl-3,5-pyridinedicarbonitrile N-methylmorpholinium salt together with 53.3 mg (0.244 mmol) of bromoacetamide and 54.7 mg (0.65 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is initially purified by preparative HPLC. The isolated fraction is reconcentrated under reduced pressure and the residue is purified by preparative thin-layer chromatography.
Yield: 14 mg (18.2% of theory) of product
Mass spectrum: molecular mass calculated: 471, found [M+H]+=472.1
82 mg (0.164 mmol) of 2-amino-4-{4-(piperidinosulfonyl)phenyl}-6-sulfanyl-3,5-pyridinedicarbonitrile N-methylmorpholinium salt together with 53.5 mg (0.246 mmol) of bromoacetamide and 55 mg (0.65 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15 μ 10×20 mm
Wavelength: 220 mm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 42.8 mg (57.2% of theory) of product
NMR [400 MHz, DMSO-d6]: 1.4 m (2H), 1.6 m (4H), 3.0 tr (4H), 3.9 s (2H), 7.25 s (1H), 7.5 s (1H), 7.8 d (2H), 7.9 d (2H), 8.1 s broad (2H)
90 mg (0.179 mmol) of 2-amino-4-{4-(morpholinosulfonyl)phenyl}-6-sulfanyl-3,5-pyridinedicarbonitrile N-methylmorpholinium salt together with 58.5 mg (0.269 mmol) of bromoacetamide and 60 mg (0.71 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 mm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 43.7 mg (53.2% of theory) of product
NMR[400 MHz, DMSO-d6]: 2.9 tr (4H), 3.65 tr (4H), 3.9 s (2H), 7.25 s (1H), 7.5 s (1H), 7.85 d (2H), 7.95 d (2H), 8.15 s broad (2H)
135 mg (0.316 mmol) of 2-[4-(2-amino-3,5-dicyano-6-sulfanyl-4-pyridinyl)-phenoxy]acetic acid N-methylmorpholinium salt together with 103.3 mg (0.474 mmol) of bromoacetamide and 106.1 mg (1.263 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is prepurified by preparative HPLC. The isolated fraction is reconcentrated under reduced pressure and the residue is purified by preparative thin-layer chromatography.
Yield: 14 mg (11.6% of theory) of product
Mass spectrum: molecular mass calculated: 383, found [M+Na]+=406.2
72 mg (0.18 mmol) of 2-[4-(2-amino-3,5-dicyano-6-sulfanyl-4-pyridinyl)benzoic acid N-methylmorpholinium salt together with 59.2 mg (0.27 mmol) of bromoacetamide and 60.9 mg (0.72 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is prepurified by preparative HPLC. The isolated fraction is reconcentrated under reduced pressure and the residue is purified by preparative thin-layer chromatography.
Yield: 11 mg (17.2% of theory) of product
Mass spectrum: molecular mass calculated: 353, found [M+H]+=353.9
89 mg (0.216 mmol) of methyl 4-(2-amino-3,5-dicyano-6-sulfanyl-4-pyridinyl)-benzoate N-methylmorpholinium salt together with 70.7 mg (0.324 mmol) of bromoacetamide and 72.7 mg (0.86 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution is purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 mm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 40.4 mg (50.8% of theory) of product
NMR [400 MHz, DMSO-d6]: 3.9 s (2H), 7.25 s (1H), 7.5 s (1H), 7.7 d (2H), 8.1 d (2H), 8.1 s broad (2H)
44 mg (0.11 mmol) of N-[4-(2-amino-3,5-dicyano-6-sulfanyl-4-pyridinyl)phenyl]-acetamide N-methylmorpholinium salt together with 35 mg (0.16 mmol) of bromoacetamide and 36 mg (0.43 mmol) of NaHCO3 are stirred in 0.5 ml of DMF overnight. After filtration, the reaction solution was purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 mm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 400 μl of DMF solution
Yield: 18.3 mg (46.6% of theory) of product
NMR [400 MHz, DMSO-d6]: 2.1 s (3H), 3.9 s (2H), 7.25 s (1H), 7.5 d (3H), 7.7 d (2H), 8.0 s broad (2H), 10.25 s (1H)
26.8 mg (0.1 mmol) of 2-amino-4-(4-hydroxyphenyl)-6-sulfanyl-3,5-pyridinedicarbonitrile are dissolved in 0.2 ml of dimethylformamide. 20 mg (0.238 mmol) of solid sodium bicarbonate are added, followed by a solution of 18.74 mg (0.15 mmol) of 2-bromoethanol in 0.06 ml of dimethylformamide. The reaction mixture is shaken overnight and, after filtration, purified by preparative HPLC.
HPLC Conditions:
Column: GROM-SIL 120 ODS 4 HE 5μ 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 300 μl of DMSO solution
Retention time: 3.97 min
Yield: 14.1 mg (45.1% of theory)
Mass spectrum: molecular mass calculated: 312, found [M+H]+=313
1. Step:
32.6 g (0.2 mol) of 4-acetaminobenzaldehyde and 13.74 g (0.208 mol) of malononitrile are initially charged in 140 ml of ethanol, and 24 drops of piperidine are added. The mixture is stirred at reflux for 30 min. After cooling, the crystals are filtered off with suction and dried.
Yield: 38.6 g (90.6% of theory) of product
Mass spectrum: molecular mass calculated: 211, found [M+H]+=212
2. Step
19 g (0.09 mol) of N-[4-(2,2-dicyanovinyl)phenyl]acetamide, 5.95 g (0.09 mol) of malononitrile and 9.91 g (0.09 mol) of thiophenol are initially charged in 120 ml of ethanol, and 0.4 ml of triethylamine are added. The mixture is stirred at reflux for 2 h, during which the product crystallizes. After cooling, the product is filtered off with suction and dried under reduced pressure.
Yield: 10.25 g (29.6% of theory) of product
Mass spectrum: molecular mass calculated: 385, found [M+H]+=386
3. Step
Under argon, 1.16 g (3 mmol) of N-{4-[2-amino-3,5-dicyano-6-(phenylsulfanyl)-4-pyridinyl]phenyl}-acetamide are dissolved in 10 ml of DMF, 0.78 g (10 mmol) of sodium sulfide are added and the mixture is stirred at 80° C. for 2 h. 20 ml of 1N HCl are then added and the resulting crystals are filtered off with suction and dried under reduced pressure.
Yield: 428 mg (46.1% of theory) of product
Mass spectrum: molecular mass calculated: 309, found [M+H]+=310.1
4. Step
309 mg (1 mmol) of N-[4-(2-amino-3,5-dicyano-6-sulfanyl-4-pyridinyl)phenyl]-acetamide, 241 mg (1 mmol) of 2-(bromomethyl)-1H-imidazole hydrobromide and 336 mg (4 mmol) of NaHCO3 in 2 ml of DMF are stirred at RT. After 2 h, 4 to 5 ml of water are added and the beige crystals are filtered off with suction and dried under reduced pressure. The crystals (310 mg) are dissolved in DMSO and purified by prep. HPLC using 9 injections. The corresponding fraction is concentrated under reduced pressure and the crystalline residue is suspended in water, filtered off with suction and dried under reduced pressure.
HPLC Conditions:
Column: Kromasil 100 C18 5 μm 50×20 mm
Precolumn: GROM-SIL ODS 4 HE 15μ 10×20 mm
Wavelength: 220 nm
Flow rate: 25 ml/min
Gradient: A=acetonitrile+0.1% trifluoroacetic acid
Injection volume: 500 μl of DMSO solution
Retention time: 3.6 min
Yield: 234 mg (60% of theory) of product
Mass spectrum: molecular mass calculated: 389, found [M+H]+=390.1
1H-NMR (300 MHz, DMSO-d6): δ=2.1 s (3H), 4.7 s (1H), 7.4 d (2H), 7.55 s (1H), 7.7 d (2H), 8.1 s broad (2H), 10.25 s (1H), 14.2 s broad (1H)
The compounds listed in the tables below (examples A 1 to A 377, A 378 to A 413 and B 1 to B 375) were prepared analogously to the procedures given above. Identity and purity of the compounds was demonstrated by LC-MS.
The compounds of examples A 1 to A 413 were either isolated as crystals or, if they did not crystallize directly from the reaction solution, purified by preparative HPLC.
The compounds of examples B 1 to B 375 were prepared on a 10 μmol scale, analogously to the procedures above. These compounds were purified and identified by a preparative HPLC-MS system.
In the tables below, structures having a group —N— are in each case understood to contain a group —NH— and structures having a group —N are in each case to be understood as containing a group —NH2.
| Number | Date | Country | Kind |
|---|---|---|---|
| 199 47 154 | Oct 1999 | DE | national |
This application is a divisional of U.S. application Ser. No. 10/110,284 filed Aug. 19, 2002 which is now U.S. Pat. No. 7,135,486, which is a §371 of International Application No. PCT/EP00/09153 filed Sep. 19, 2000.
| Number | Date | Country | |
|---|---|---|---|
| 20060264432 A1 | Nov 2006 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 10110284 | US | |
| Child | 11359927 | US |
