Patent Application
20130196050
1. Field of the Invention
The present invention relates to novel compounds having a salty taste enhancing effect. Moreover, the present invention relates to salty taste enhancer comprising such a compound which is used for food and drink, and the like.
2. Discussion of the Background
Excess intake of sodium chloride is one cause of elevated blood pressure and is considered to cause cerebral apoplexy and heart disease. To prevent this, reduction of the intake of sodium chloride has been recommended. However, reduced-salt foods having a reduced amount of sodium chloride taste plain and markedly reduce taste quality. To improve plain taste due to the reduction of salt, a method including addition of sodium glutamate and spice is known (see, e.g., The Japanese Journal of Taste and Smell Research, vol. 14, No. 3, page 447-450, 2007). While sodium glutamate and spice enhance the flavor, they are not sufficient for the effect of enhancing the salty taste itself.
Furthermore, a method of replacing a part of sodium chloride with a sodium chloride alternative such as potassium salt, ammonium salt, basic amino acid, salty taste peptide and the like, has also been reported. For example, a method of reducing a bitter taste of potassium chloride by using potassium chloride and carrageenan in combination (see, e.g., JP-A-H4-262758), a production method of potassium chloride-containing fermentation food (see, e.g., JP-A-2007-289145) and the like. Moreover, a substance free of a sodium chloride taste in itself but potentiates a salt taste when co-used with sodium chloride is also known. For example, a method of adding a saturated aliphatic monocarboxylic acid having 3 to 8 carbon atoms to a sodium chloride-containing food and drink at a proportion of 0.01 to 1 wt % based on the weight of sodium chloride and the like is known (see, e.g., JP-A-H5-184326). However, they are not satisfactory methods in terms of strength of the salty taste and taste quality.
Therefore, a reduced-salt food superior in taste property, a salty taste enhancer which is superior in the taste quality or strong, or further, a novel compound capable of enhancing the salty taste is still demanded.
On the other hand, various studies of the receptive mechanism of salty taste have been undertaken, though many aspects remain to be clarified.
ENaC (epithelial sodium channel) is a voltage-independent, amiloride-sensitive sodium channel present in the cellular membrane, which is an ion channel that functions when three kinds of subunits (α or δ subunit, β subunit and γ subunit) are bonded. ENaC is known as an influx pathway of sodium ion in many epithelial tissues (see Palmer L G (1987). “Ion selectivity of epithelial Na channels”, J. Membr. Biol., 96: 97-106 and Lazdunski M, Waldmann R, Champigny G, Bassilana F, Voilley N (1995), “Molecular cloning and functional expression of a novel amiloride-sensitive Na+ channel”. J. Biol. Chem. 270, (46): 27411-27414).
ENaC is one of the proteins particularly studied in relation to the salty taste. Nevertheless, the involvement thereof in the salty taste acceptance of humans has not been clarified as yet. While the involvement thereof in the salty taste acceptance of rodents has been acknowledged (see Chandrashekar, J. et al. The cells and peripheral representation of sodium taste in mice, Nature, (2010), 464, 297-302), there are negative opinions on that in the salty taste acceptance of humans. In fact, while Lu, M., et al. Small Molecule Activator of the Human Epithelial Sodium Channel, Journal of Biological Chemistry (2008), 283(18), 11981-11994 describes that S3969 [N-(2-hydroxyethyl)-4-methyl-2-(4-methyl-1H-indol-3-ylthio)pentanamide] is a stimulant (activator) of ENaC that acts on β subunit, it merely suggests that a salty taste may have a stimulant action on rodents, and is completely silent on how it is actually tasted by humans during eating, even a possibility thereof.
While Yamazaki Y., Kawano Y., Yamanaka A., and Maruyama S., Bioorganic & Medicinal Chemistry Letters, 19, (2009) 4178-4182 and JP-A-H8-53332 and the like describe that particular serotonin derivatives have a whitening effect and the like, the action of such serotonin derivatives on taste of sense is not known at all.
Accordingly, it is one object of the present invention to provide novel compounds having a strong salty taste enhancing effect.
It is another object of the present invention to provided novel salty taste enhancers which contain such a compound, and the like.
These and other objects, which will become apparent during the following detailed description, have been achieved by the inventors' discovery that compounds of the following structure have a strong salty taste enhancing effect.
Accordingly, the present invention provides the following.
wherein
wherein
wherein
wherein
Q is a group represented by
wherein
Hal is a halogen atom; and
* means a bonding site to X, and
X is a covalent bond.
Q is a group represented by
wherein
wherein
wherein
wherein
wherein * means a bonding site to X, and
wherein
wherein
wherein
wherein
wherein
wherein
wherein
wherein
R13 and R14 are each independently a hydrogen atom, a hydroxyl group, an alkyl group having 1 to 3 carbon atoms or an alkoxy group having 1 to 3 carbon atoms,
(2) a compound wherein
wherein
wherein
Q″ is a group represented by
wherein
wherein * means a bonding site to X, and
R9′ is a hydrogen atom,
or a salt thereof.
or a salt thereof.
The compounds of the present invention are expected to show a strong salty taste enhancing effect and can be used for a salty taste enhancer for food and drink, and the like.
A more complete appreciation of the invention and many of the attendant advantages thereof will be readily obtained as the same become better understood by reference to the following detailed description when considered in connection with the accompanying drawings, wherein:
The terms used in the present specification are explained below.
Examples of the “halogen atom” include a chlorine atom, a bromine atom, a fluorine atom and an iodine atom.
Examples of the “alkyl group having 1 to 3 carbon atoms” include a methyl group, an ethyl group, a propyl group, an isopropyl group and the like.
Examples of the “alkyl group having 1 to 6 carbon atoms” include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group, a neopentyl group, a 1-ethylpropyl group, a 1,1-dimethylpropyl group, a 2-methylbutyl group, a hexyl group, an isohexyl group, a 1,1-dimethylbutyl group, a 2,2-dimethylbutyl group, a 3,3-dimethylbutyl group, a 2-ethylbutyl group and the like.
Examples of the “alkoxy group having 1 to 3 carbon atoms” include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group and the like.
Examples of the “cycloalkyl group having 3 to 7 carbon atoms” include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group and the like.
Examples of the “alkoxy-carbonyl group having 2 to 4 carbon atoms” include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group and the like.
Examples of the “alkoxy-carbonyl group having 2 to 5 carbon atoms” include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group, a butoxycarbonyl group and the like.
Examples of the “aralkyloxycarbonyl group” include a benzyloxycarbonyl group, a phenethyloxycarbonyl group, a naphthylmethyloxycarbonyl group (a 1-naphthylmethyloxycarbonyl group, a 2-naphthylmethyloxycarbonyl group), a biphenylylmethyloxycarbonyl group and the like.
Examples of the “alkylene group having 1 to 3 carbon atoms” include a methylene group, an ethylene group, a propylene group, —CH(CH3)—, —CH(CH3)—CH2—, —CH2—CH(CH3)—, —C(CH3)2—, —CH(C2H5)— and the like.
Examples of the “alkylenedioxy group having 1 to 3 carbon atoms” include a methylenedioxy group, an ethylenedioxy group, a propylenedioxy group, —O—CH(CH3)—O—, —O—CH(CH3)—CH2—O—, —O—CH2—CH(CH3)—O—, —O—C(CH3)2—O—, —O—CH(C2H5)—O— and the like.
Each moiety in the above-mentioned formula (I) is explained below.
Q is
(1) a group represented by the following formula:
wherein
The “alkyl group having 1 to 3 carbon atoms” of the “alkyl group having 1 to 3 carbon atoms which is optionally substituted by 1 to 3 halogen atoms” for R1, R2, R3, R4 or R5 is preferably a methyl group.
The “alkyl group having 1 to 3 carbon atoms which is optionally substituted by 1 to 3 halogen atoms” for R1, R2, R3, R4 or R5 is preferably an alkyl group having 1 to 3 carbon atoms (preferably a methyl group) which is optionally substituted by 1 to 3 fluorine atoms, more preferably a trifluoromethyl group.
The “alkoxy group having 1 to 3 carbon atoms” for R1, R2, R3, R4 or R5 is preferably a methoxy group.
The “alkylenedioxy group having 1 to 3 carbon atoms” formed by R1 and R2 in combination or R2 and R3 in combination is preferably a methylenedioxy group.
The “cycloalkyl group having 3 to 7 carbon atoms” of the “cycloalkyl group having 3 to 7 carbon atoms which is optionally substituted by 1 to 3 alkyl groups having 1 to 3 carbon atoms” for Q is preferably a cyclohexyl group.
The “cycloalkyl group having 3 to 7 carbon atoms” is optionally substituted by 1 to 3 substituents selected from an alkyl group having 1 to 3 carbon atoms (preferably a methyl group, an isopropyl group).
The “cycloalkyl group having 3 to 7 carbon atoms which is optionally substituted by 1 to 3 alkyl groups having 1 to 3 carbon atoms” for Q is preferably a cyclohexyl group optionally substituted by 1 to 3 alkyl groups having 1 to 3 carbon atoms (preferably a methyl group, an isopropyl group), more preferably a cyclohexyl group optionally substituted by 1 or 2 alkyl groups having 1 to 3 carbon atoms (preferably a methyl group, an isopropyl group), particularly preferably a 2-isopropyl-5-methylcyclohexyl group.
The “alkyl group having 1 to 6 carbon atoms” of the “alkyl group having 1 to 6 carbon atoms which is optionally substituted by 1 to 3 substituents selected from an amino group and a hydroxyl group” for Q is preferably an ethyl group, a tert-butyl group, an isopentyl group or a neopentyl group.
The “alkyl group having 1 to 6 carbon atoms which is optionally substituted by 1 to 3 substituents selected from an amino group and a hydroxyl group” for Q is preferably an alkyl group having 1 to 6 carbon atoms (preferably an ethyl group, a tert-butyl group, an isopentyl group, a neopentyl group) optionally substituted by 1 or 2 (preferably 1) substituents selected from an amino group and a hydroxyl group, more preferably a tert-butyl group, a neopentyl group, a 2-hydroxyethyl group or a 1-amino-3-methylbutyl group.
Q is preferably a group represented by
wherein
Hal is a halogen atom; and
* means a bonding site to X.
In another embodiment, Q is preferably a group represented by
wherein
In another embodiment, Q is
preferably
(1) a group represented by the following formula:
wherein
In another embodiment, Q is
preferably a group represented by
wherein
wherein * means a bonding site to X.
In another embodiment, Y is preferably —CO—NH—.
The “alkyl group having 1 to 3 carbon atoms” for R8 is preferably a methyl group.
The “alkylene group having 1 to 3 carbon atoms” formed by R8 and R11 in combination is preferably a methylene group.
Z is —CHR9—, —CH2—CR9R10—, —CR9R10—CH2—, —CH2—CH2—CHR9—, —CH═CR9— or —CR9═CH—
The “alkyl group having 1 to 3 carbon atoms” of the “alkyl group having 1 to 3 carbon atoms which is optionally substituted by hydroxyl group(s)” for R9 or R10 is preferably a methyl group.
The “alkoxy-carbonyl group having 2 to 4 carbon atoms” for R9 or R10 is preferably a methoxycarbonyl group or an ethoxycarbonyl group.
The “aralkyloxycarbonyl group” for R9 or R10 is preferably a benzyloxycarbonyl group.
The “alkoxy-carbonyl group having 2 to 5 carbon atoms” of the “amino group optionally substituted by alkoxy-carbonyl group(s) having 2 to 5 carbon atoms” for R9 or R10 is preferably a tert-butoxycarbonyl group.
The “alkyl group having 1 to 3 carbon atoms” of the “carbamoyl group optionally substituted by alkyl group(s) having 1 to 3 carbon atoms wherein the alkyl group is optionally substituted by 1 to 3 substituents selected from a hydroxyl group, a carboxyl group and a phenyl group” for R9 or R10 is preferably an ethyl group.
Z is preferably —CHR9—, —CH2—CR9R10—, —CH2—CH2—CHR9— or —CH═CR9—, more preferably —CHR9—, —CH2—CR9R10—, —CH2—CH2—CHR9— or —CH═CH—
In another embodiment, Z is
preferably —CH2—CHR9′— or —CH2—CH2—CHR9′—
Ar is
(1) a group represented by the following formula:
wherein
wherein
The “alkylene group having 1 to 3 carbon atoms” formed by R11 and R8 in combination is preferably a methylene group.
The “alkyl group having 1 to 3 carbon atoms” for R12, R13, R13a, R14 or R14a is preferably a methyl group.
The “alkoxy group having 1 to 3 carbon atoms” for R15, R15a, R15b, R16 or R16a is preferably a methoxy group.
The “alkylenedioxy group having 1 to 3 carbon atoms” formed by R15 and R16 in combination is preferably a methylenedioxy group.
R13a, R14a, R15a, R15b and R16a are preferably hydrogen atoms.
Ar is preferably
(1) a group represented by the following formula:
wherein each symbol is as defined above, or
(2) a group represented by the following formula;
wherein each symbol is as defined above.
Ar is preferably
(1) a group represented by the formula (III′)
In another embodiment, Ar is preferably a group represented by the following formula:
wherein each symbol is as defined above.
The compound represented by the formula (I) wherein
(1) a group represented by the above-mentioned formula (II)
(1) a group represented by the formula (III′)
In another embodiment, the compound represented by the formula (I) wherein R13a, R14a, R15a, R15b and R16a are hydrogen atoms is preferable.
In another embodiment, the compound represented by the formula (I) wherein
Q is a group represented by
wherein
Hal is a halogen atom; and
* means a bonding site to X, and
X is a covalent bond,
is preferable.
In another embodiment, the compound represented by the formula (I) wherein
Q is a group represented by
wherein
In another embodiment, the compound represented by the formula (I) is preferably a compound represented by the formula (V), particularly preferably the compound wherein n is 2.
In another embodiment, a compound represented by the formula (I) is preferably a compound represented by the formula (VI), particularly preferably the compound wherein Q″ is a group represented by
wherein * means a bonding site to X, and
R9′ is a hydrogen atom.
In another embodiment, the compound represented by the formula (I) wherein
any of R1 to R5 is a sulfo group, and
is preferable.
The compound represented by the formula (I) is particularly preferably the compounds disclosed in the following Examples, particularly 2,6-dihydroxy-N-(2-(5-hydroxy-1H-indol-3-yl)ethyl)benzamide.
Of compound (I), the compound represented by the above-mentioned formula (I′) is a novel compound.
The compound represented by the formula (I′) wherein Q′ is a group represented by
(1) the above-mentioned formula (II′)
(1) a group represented by the formula (III′)
Examples of the salt of compound (I) or (I′) include salts with inorganic acid, salts with organic acid, salts with inorganic base, salts with organic base, salts with acidic or basic amino acid, and the like.
Examples of the salt with inorganic acid include hydrochloride, hydrobromide, sulfate, nitrate, phosphate and the like.
Examples of the salt with organic acid include formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, p-toluenesulfonate and the like.
Examples of the salt with inorganic base include sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt and the like.
Examples of the salt with organic base include salts with methylamine, diethylamine, trimethylamine, triethylamine, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, tris(hydroxymethyl)methylamine, dicyclohexylamine, N,N′-dibenzylethylenediamine, guanidine, pyridine, picoline, choline, cinchonine, meglumine and the like.
Examples of the salt with acidic or basic amino acid include salts with aspartic acid, glutamic acid, arginine, lysine and ornithine.
The above-mentioned salt is preferably an edible salt.
When compound (I) (including compound (I′)) or a salt thereof (hereinafter, sometimes to be abbreviated as the compound of the present invention) contains an optical isomer, a stereoisomer, a regioisomer or a rotamer, these are also encompassed in the compound of the present invention, and can be obtained as a single product according to synthetic and separation methods known per se. For example, when the compound of the present invention contains an optical isomer, an optical isomer resolved from racemate is also encompassed in the compound of the present invention. These isomers can be obtained as a single product according to synthetic methods known per se, separation methods known per se (e.g., concentration, solvent extraction, column chromatography, recrystallization, etc.), optical resolution methods known per se (e.g., fractional recrystallization, chiral column method, diastereomer method etc.) and the like.
In one embodiment of the present invention, the compound of the present invention can be added to various foods and drinks. While the food and drink are not particularly limited, examples thereof include seasonings (e.g., miso, soy sauce, baster, dashi (Japanese soup), dressing, mayonnaise, tomato ketchup etc.), soups (e.g., miso soup, Japanese-style soup (osuimono), consomme soup, egg soup, seaweed soup, potage etc.), sauces for soba, wheat noodles (udon), ramen, pasta and the like, sauces, foods of cooked rice (e.g., rice gruel, rice soup, ochazuke etc.), livestock processed products (e.g., ham, sausage, cheese etc.), confectionery and snack foods (e.g., potato chips, Japanese cracker, cookie etc.), cooked foods (e.g., boiled food, fried food, roasted food, curry etc.), drinks and the like.
The amount of the compound of the present invention to be added to food and drink is not particularly limited as long as the effect thereof can be exhibited. However, since food and drink are mixtures with various substances, the amount of the present invention that achieves a salty taste enhancing effect may vary from the amount that shows a salty taste enhancing effect using simple brine and the like. Therefore, the amount of the compound of the present invention to be added to food and drink can be determined by appropriately examining the optimal amount for each food and drink. For example, 0.000001 to 0.1 wt %, based on the total weight of the food or drink is preferable.
The compound of the present invention can be used in combination with a known salty taste alternative. Examples of the salty taste alternative include potassium chloride, organic acid, arginine, argininate, ammonium chloride and the like. These may be used alone or two or more kinds thereof may be used as a mixture.
When the compound of the present invention is added to food and drink, the food and drink produced may contain a suitable additive to the extent that the salty taste enhancing action is not prevented. For example, such food and drink can contain, in addition to the compound of the present invention, various additives generally usable for the production of food and drink, such as protein (milk protein, soybean protein etc.), inorganic salt, acid, amino acids, nucleic acid taste components, saccharides, fats, natural seasonings, spices, excipients, dye components and the like.
Examples of the inorganic salt include potassium chloride, ammonium chloride, magnesium sulfate and the like.
Examples of the acid include carboxylic acids such as ascorbic acid, fumaric acid, malic acid, tartaric acid, citric acid, lactic acid, succinic acid and the like, salts thereof and the like.
Examples of the amino acids include glutamates (e.g., sodium glutamate, potassium glutamate, calcium glutamate, ammonium glutamate, magnesium glutamate etc.), glutamic acid and the like. These have already been used as flavor enhancers for food, and all of them have umami taste derived from glutamic acid and taste property (e.g., sour taste derived from ammonium salt, etc.) characteristic of each cation. In addition, basic amino acids (e.g., lysine, arginine, histidine etc.) and salts thereof can also be used as amino acids.
Examples of the nucleic acid taste component include sodium inosinate, sodium guanylate and the like.
Examples of the saccharide include sucrose, glucose, lactose and the like.
The compound of the present invention preferably contains one or more kinds of additives selected particularly from organic acids, arginine, argininate, ammonium chloride and potassium chloride, which are known to have a salty taste enhancing effect.
The production method of the compound represented by the formula (I) or a salt thereof is not particularly limited, and it can be produced by combination of known methods. Specifically, it can be synthesized according to the following method, which is not to be construed as limitative.
The compound represented by the formula (I) wherein Y is —NR8—CO— wherein R8 is as defined above, or a salt thereof (hereinafter, to be referred to as compound (I-1)) can be produced according to the following Production Methods 1 or 2.
wherein each symbol is as defined above.
Compound (I-1) can be produced by subjecting amine component (VII) and carboxylic acid component (VIII) to a condensation reaction with a dehydrating condensing agent.
Amine component (VII) may be in the form of a salt (e.g., hydrochloride, p-toluenesulfonate etc.), and carboxylic acid component (VIII) may be in the form of a salt (e.g., dicyclohexylamine salt etc.). When amine component (VII) is in the form of a salt, the condensation reaction may be carried out by addition of a base (e.g., triethylamine etc.). While the ratio of amine component (VII) and carboxylic acid component (VIII) to be used is not limited, carboxylic acid component (VIII) is used in an amount of 0.8 to 1.2 equivalents per 1 equivalent of amine component (VII) for the reaction in good yield. The amount of the base to be used is 0.8 to 2.0 equivalents, preferably 1.0 to 1.5 equivalents, relative to amine component (VII).
The solvent to be used is not particularly limited as long as it does not react with amine component (VII) or carboxylic acid component (VIII), and examples thereof include dichloromethane (DCM), N,N-dimethylformamide (DMF), chloroform, dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP), and a mixed solvent thereof. Among them, dichloromethane and N,N-dimethylformamide are preferable. The amount of the solvent is 10 to 500-fold weight, preferably 15 to 100-fold weight, relative to amine component (VII).
The dehydrating condensing agent may be a general condensing agent used for peptide synthesis and the like, and examples thereof include N,N′-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) and the like. A condensation promoter (e.g., 1-hydroxybenzotriazole (HOBt) etc.) is also used together with the dehydrating condensing agent. The amount of the dehydrating condensing agent to be used is 1.0 to 2.0 equivalents, preferably 1.05 to 1.20 equivalents, relative to amine component (VII). The amount of the condensation promoter to be used is 0.5 to 3.0 equivalents, preferably 1.0 to 1.5 equivalents, relative to amine component (VII).
The reaction time is preferably about 3 to 24 hours which depends on the reaction temperature. The reaction temperature is preferably within the range of 5 to 35° C.
wherein each symbol is as defined above.
Compound (I-1) can be produced by converting carboxylic acid component (VIII) to acid chloride (IX), and then subjecting acid chloride (IX) and amine component (VII) to a condensation reaction in the presence of a base.
Acid chloride (IX) can be obtained by reacting carboxylic acid component (VIII) with oxalyl chloride, thionyl chloride or the like according to a conventional method. The amount of the oxalyl chloride, thionyl chloride or the like to be used is 0.8 to 10.0 equivalents, preferably 1.0 to 2.0 equivalents, relative to carboxylic acid component (VIII). The reaction temperature is generally 10.0 to 50.0° C., preferably 0 to 35.0° C., and the reaction time is generally 1.0 to 20.0 hours, preferably 3.0 to 16.0 hours.
The reaction can be carried out by reacting acid chloride (IX) with amine component (VII) in the presence of a base (e.g., triethylamine, sodium hydroxide etc.). While the ratio of amine component (VII) and acid chloride (XI) to be used is not limited, acid chloride (IX) is used in an amount of 0.8 to 1.2 equivalents relative to amine component (VII), for the reaction in good yield. The amount of the base to be used is 0.8 to 3.0 equivalents, preferably 1.0 to 1.5 equivalents, relative to amine component (VII). Examples of the solvent to be used include those used in the above-mentioned Production
The reaction time is preferably about 3 to 24 hours which depends on the reaction temperature. The reaction temperature is preferably within the range of 5 to 35° C.
The compound represented by the formula (I) wherein Y is —CO—NH—, or a salt thereof (hereinafter, to be referred to as compound (I-2)) can be produced according to the following Production Methods 3 or 4.
wherein each symbol is as defined above.
Compound (I-2) can be produced by subjecting amine component (XI) and carboxylic acid component (X) to the reaction similar to that in Production Method 1.
Compound (I-2) can be produced by subjecting amine component (XI) and carboxylic acid component (X) to the reaction similar to that in Production Method 2.
The compound represented by the formula (I) wherein Y is —NH—CO—O—, or a salt thereof (hereinafter, to be referred to as compound (I-3)) can be produced according to the following Production Method 5.
Compound (I-3) can be obtained by reacting amine component (XII) with chloroformate (XIII) (e.g., benzyl chloroformate etc.) or dicarbonate (XIV) (e.g., dialkyl dicarbonate) in the presence of a base under general conditions. Examples of the base include triethylamine, diisopropylamine, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogen carbonate and the like.
While the ratio of amine component (XII) and chloroformate (XIII) or dicarbonate (XIV) to be used is not limited, chloroformate (XIII) or dicarbonate (XIV) is used in an amount of 0.9 to 2.0 equivalents per 1 equivalent of amine component (XII), for the reaction in good yield. The amount of the base to be used is 1.5 to 3.0 equivalents, preferably 2.0 to 2.5 equivalents, relative to amine component (XII).
The solvent to be used is not particularly limited as long as it does not rapidly react with amine component (XII), chloroformate (XIII) or dicarbonate (XIV), and examples thereof include 1,4-dioxane, dichloromethane, THF, water and the like.
The reaction time is generally 1.0 to 25 hours, preferably 5.0 to 18 hours, and the reaction temperature is generally 0 to 50° C., preferably 15 to 35° C.
In each of the above-mentioned reactions, when desired, each substituent can be converted by known deprotection, acylation reaction, alkylation reaction, hydrogenation reaction, oxidation reaction, reduction reaction, carbon chain extension reaction or substituent exchange reaction alone or a combination of two or more thereof.
In each of the above-mentioned reactions, when the starting material compound has an amino group, a carboxyl group, a hydroxy group or a carbonyl group as a substituent, a protecting group generally used in peptide chemistry and the like may be introduced into these groups. By removing the protecting group as necessary after the reaction, the objective compound can be obtained.
The removal of the above-mentioned protecting group can be performed according to a known method, for example, the method described in Protective Groups in Organic Synthesis, John Wiley and Sons (1980) or and the like.
The obtained compound represented by the formula (I) or a salt thereof can be isolated and purified according to a conventional method. For example, when it is purified by crystallization, ethyl acetate, ethanol, methanol, diethyl ether, chloroform, dichloromethane, n-hexane and a mixed solvent thereof can be used as a solvent. Preparative thin layer chromatography (PTLC) or silica gel column chromatography can be employed as purification using chromatography. In this case, solvents exemplified for the above-mentioned crystallization can be used as an eluent.
Other features of the invention will become apparent in the course of the following descriptions of exemplary embodiments which are given for illustration of the invention and are not intended to be limiting thereof.
The structures of the compounds synthesized in the following Examples were determined by nuclear magnetic resonance spectrum (Bruker AVANCE 400).
2,6-Dihydroxybenzoic acid (0.7706 g, 5.0 mmol) was dissolved in N,N-dimethylformamide (15 mL), and 1-hydroxybenzotriazole monohydrate (HOBt.H2O) (1.5314 g, 10.0 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (1.9170 g, 10.0 mmol) were added thereto, and the mixture was stirred at room temperature for 30 minutes. Then, (L)-tryptophan methyl ester hydrochloride (1.5283 g, 6.0 mmol) and triethylamine (1.66 mL, 12.0 mmol) were added thereto, and the mixture was stirred overnight at room temperature. After completion of the reaction, the solvent was evaporated, and the residue was diluted with ethyl acetate. The mixture was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 10% saturated aqueous sodium hydrogen carbonate (twice) and saturated brine (once). The organic layer was dried over magnesium sulfate, and filtered, and the solvent was evaporated. The obtained crude product was purified by silica gel column chromatography to give the objective compound (0.6036 g, 1.70 mmol) as a white solid.
The compounds of Examples 2, 5, 10, 11, 14-17, 20, 21, and 24-29 shown in the following Table 1 were also synthesized in the same manner as in Example 1. The compound of Example 20 was synthesized in the same manner as in Example 21, and it was confirmed that the objective compound was produced by TLC (silica gel 60 F254, manufactured by Merck; eluent, hexane:ethyl acetate=2:1).
(S)-Methyl 2-(2,6-dihydroxybenzamido)-3-(1H-indol-3-yl)propanoate obtained in Example 1 (0.5508 g, 1.55 mmol) was dissolved in tetrahydrofuran (8 mL), and 1M lithium hydroxide aqueous solution (3.1 mL) was added thereto, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the pH of the mixture was adjusted to about 2 with 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The obtained organic layer was washed with saturated brine, dried over magnesium sulfate, and filtered, and the solvent was evaporated. The obtained crude product was purified by silica gel column chromatography to give the objective compound as a white solid.
The compounds of Examples 4, 9, 12, 13, 18, and 19 shown in the following Table 1 were also synthesized in the same manner as in Example 3.
Salicylic acid (0.2762 g, 2.0 mmol), HOBt.H2O (0.3982 g, 2.6 mmol), and EDCI.HCl (0.4984 g, 2.6 mmol) (6 mL) were dissolved in N,N-dimethylformamide, and serotonin hydrochloride (0.5530 g, 2.6 mmol) and triethylamine (0.720 mL, 5.2 mmol) were added thereto, and the mixture was stirred overnight at room temperature. After completion of the reaction, the solvent was evaporated, and the residue was diluted with ethyl acetate. The mixture was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 10% saturated aqueous sodium hydrogen carbonate (twice) and saturated brine (once). The organic layer was dried over magnesium sulfate, and filtered, and the solvent was evaporated. The obtained crude product was purified by silica gel column chromatography to give the objective compound (0.4609 g, 1.56 mmol).
The compounds of Examples 7 and 8 shown in the following Table 1 were also synthesized in the same manner as in Example 6.
To tert-butyl (S)-1-(2,6-dihydroxy phenylcarbamoyl)-2-(1H-indol-3-yl)ethylcarbamate obtained in Example 21 (0.520 g, 1.27 mmol), was added 4M hydrogen chloride-dioxane solution (16 mL), and the mixture was reacted overnight at room temperature. After completion of the reaction, the solvent was evaporated, and the pH of the residue was adjusted to about 11 with 1M aqueous sodium hydroxide solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, and filtered, and the solvent was evaporated. The obtained crude product was purified by silica gel column chromatography to give the objective compound (0.0891 g, 0.29 mmol).
The compound of Example 23 shown in the following Table 1 was also synthesized in the same manner as in Example 22, and it was confirmed that that the objective compound was produced by TLC (silica gel 60 F254; eluent, dichloromethane:methanol=9:1).
2,6-Dihydroxybenzoic acid (0.312 g, 2 mmol) and serotonin hydrochloride (0.430 g, 2 mmol) were dissolved in DMF (25 ml), and the solution was cooled to 0° C. Triethylamine (0.31 ml, 2.2 mmol), HOBt.H2O (0.346 g, 2.2 mmol), and EDCI.HCl (0.439 g, 2.2 mmol) were added thereto, and the mixture was stirred overnight at ambient temperature. The next day, the reaction progress was confirmed by thin layer chromatography (TLC), and the solvent was evaporated. To the residue were added ethyl acetate and water, and the obtained ethyl acetate layer was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 5% aqueous sodium hydrogen carbonate solution (twice) and saturated brine (once), and dried over magnesium sulfate. The magnesium sulfate was removed, and the ethyl acetate was evaporated under reduced pressure to give an amorphous compound. The amorphous compound was purified by preparative thin layer chromatography (PTLC), and recrystallized from hexane to give the objective compound (0.093 g, 0.30 mmol, 14.9%).
The compounds of Examples 32-34, 36, 37, 42, 44, 46-48, 62, 64, 67, 69, 84, 85, 87, 88, 90, 92, 93, 95, 96, 98, 104-107, 109, 111, 112, 114, 116, 118, 120, 122, 124, and 125 shown in the following Table 1 were also synthesized in the same manner as in Example 35.
2,3-Dihydroxybenzoic acid (0.663 g, 2 mmol) and tryptophan benzyl ester hydrochloride (0.309 g, 2 mmol) were dissolved in DMF (40 ml), and the solution was cooled to 0° C. Triethylamine (0.31 ml, 2.2 mmol), HOBt.H2O (0.341 g, 2.2 mmol) and EDCI.HCl (0.426 g, 2.2 mmol) were added thereto, and the mixture was stirred overnight at ambient temperature. The next day, the reaction progress was confirmed by TLC, and the solvent was evaporated. To the residue were added ethyl acetate and water, and the obtained ethyl acetate layer was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 5% aqueous sodium hydrogen carbonate solution (twice) and saturated brine (once), and dried over magnesium sulfate. The magnesium sulfate was removed, and the ethyl acetate was evaporated under reduced pressure. The residue was purified by PTLC, and recrystallized from hexane. The obtained compound was dissolved in ethanol, 5% palladium carbon (300 mg) was added thereto, and the mixture was stirred overnight under hydrogen. The next day, the palladium carbon was removed, and the residue was concentrated. The residue was recrystallized from water to give the objective compound (0.039 g, 0.11 mmol, 5.8%).
The compounds of Examples 38, 40, 41, 43, 45, 63, 65, 66, 68, 70-83, 94, 97, 99, 101, and 102 shown in the following Table 1 were also synthesized in the same manner as in Example 39.
Magnesium (668 mg) and anhydrous THF (10 mL) were stirred under argon. Iodine was added thereto, and the mixture was stirred for additional 5 minutes. (−)-Menthyl chloride (4.2 g, 240 mmol) was added thereto, and the mixture was heated with reflux. The completion of the reaction was confirmed by the change of the color of the solution from brown to colorless.
The reaction mixture was charged into 200 ml of two-necked flask, dry ice (9 g) was added thereto, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with 1N hydrochloric acid (25 mL), and the mixture was extracted with diethyl ether (25 mL×3). The organic layer was washed with water, and extracted with 1N sodium hydroxide (25 mL×3). The aqueous layer was neutralized with 2N hydrochloric acid (40 mL), and extracted with diethyl ether (25 mL×3). The organic layer was dried over sodium sulfate, and the sodium sulfate was removed by filtration. The solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give (−)-menthyl carboxylic acid (1.68 g, 9.1 mmol).
To the (−)-menthyl carboxylic acid (150 mg, 0.81 mmol) was added thionyl chloride (1 mL) under argon, and the mixture was stirred at 80° C. for 2 hours. The reaction mixture was concentrated, and the residue was cooled in ice bath, and dissolved in dichloromethane (4 mL). Tryptamine (130 mg, 0.81 mmol) and triethylamine (0.11 ml, 0.8 mmol) were added thereto, and the mixture was stirred overnight. The next day, to the mixture was added dichloromethane (6 mL), the mixture was washed with 5% aqueous citric acid solution, and the organic layer was concentrated. The residue was purified by column chromatography, and the solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give the objective compound (145 mg, 0.44 mmol, 55%).
The compounds of Examples 51, 52, and 60 shown in the following Table 1 were also synthesized in the same manner as in Example 50.
The compound obtained in Example 52 (900 mg, 1.95 mmol) was dissolved in ethanol (5 mL), 5% palladium carbon (100 mg) was added thereto, and the mixture was stirred overnight under hydrogen. The next day, the palladium carbon was removed, and the residue was concentrated. The residue was dissolved in ethyl acetate, and the solution was purified by column chromatography. The solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give the objective compound (702 mg, 1.89 mmol, 97%).
Tryptamine (200 mg, 1.2 mmol) was dissolved in anhydrous 1,4-dioxane (4 mL) under argon, triethylamine (0.35 mL, 2.5 mmol) was added thereto, and then di-tert-butyldicarbonate (300 mg, 1.4 mmol) was added thereto, and the mixture was stirred overnight. The next day, the solvent was evaporated under reduced pressure, and ethyl acetate and water were added thereto. The obtained organic layer was dried over sodium sulfate, and the sodium sulfate was removed by filtration. The solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give the objective compound (107 mg, 0.41 mmol, 34%).
Tryptamine (199 mg, 1.2 mmol) was dissolved in dichloromethane (4 mL), and diisopropylethylamine (0.44 mL) and benzyl chloroformate (0.2 ml) were added thereto, and the mixture was stirred overnight. The next day, water was added thereto. The organic layer was dried over sodium sulfate, and the sodium sulfate was removed by filtration. The filtrate was purified by silica gel column, and the solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give the objective compound (155 mg, 0.53 mmol, 43%).
The compounds of Examples 54-57 shown in the following tables were also synthesized in the same manner as in Example 53, 58, 59, and 39.
To tert-butylcarboxylic acid (0.2 ml, 1.6 mmol) was added thionyl chloride (1 mL) under argon, and the mixture was stirred at 80° C. for 2 hours. The reaction mixture was concentrated, and the residue was cooled in ice bath, and dissolved in dichloromethane (6 mL). L-Tryptophan benzyl ester hydrochloride (519 mg, 1.6 mmol) and triethylamine (0.3 mL) were added thereto, and the mixture was stirred overnight. The next day, dichloromethane (4 mL) was added thereto, and the mixture was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 5% aqueous sodium hydrogen carbonate solution (twice) and saturated brine (once), and dried over sodium sulfate. The organic layer was concentrated, and the residue was purified by column chromatography, and the solvent was evaporated under reduced pressure, and completely removed using a vacuum pump to give the objective compound (241 mg, 0.73 mmol, 47%).
2,6-Dihydroxyaniline (0.251 g, 2 mmol) and 3,4-methylene dioxycinnamic acid (0.387 g, 2 mmol) were dissolved in DMF (20 ml), and the solution was cooled to 0° C. HOBt.H2O (0.337 g, 2.2 mmol) and EDCI.HCl (0.421 g, 2.2 mmol) were added thereto, and the mixture was stirred overnight at ambient temperature. The next day, the reaction progress was confirmed by TLC, and the solvent was evaporated. To the residue were added ethyl acetate and water, and the obtained ethyl acetate layer was washed successively with 5% aqueous citric acid solution (twice), saturated brine (once), 5% aqueous sodium hydrogen carbonate solution (twice) and saturated brine (once), and dried over magnesium sulfate. The magnesium sulfate was removed, and the ethyl acetate was evaporated under reduced pressure. The residue was recrystallized from hexane. The obtained compound was dissolved in methanol, palladium hydroxide (60 mg) was added thereto, and the mixture was stirred at 40° C. for 7 hours under hydrogen. The palladium hydroxide was removed, and the residue was concentrated under reduced pressure. The precipitated crystals were removed, and the residue was purified by PTLC to give the objective compound (0.111 g, 0.37 mmol, 18.5%) as crystals.
The compounds of Examples 89 and 91 shown in the following Table 1 were also synthesized in the same manner as in Example 86.
2-Aminobenzenesulfonic acid (0.512 g, 3 mmol) and Z-homophenylalanine (0.940 g, 3 mmol) were dissolved in DMF (20 ml), and the solution was cooled to 0° C. Triethylamine (0.46 ml, 3.3 mmol), HOBt.H2O (0.505 g, 3.3 mmol), and EDCI.HCl (0.636 g, 3.3 mmol) were added thereto, and the mixture was stirred overnight at ambient temperature. The next day, the reaction progress was confirmed by TLC, and the solvent was evaporated. To the residue were added ethyl acetate and water, and the obtained ethyl acetate layer was washed successively with 5% aqueous citric acid solution (twice) and saturated brine (once), and dried over magnesium sulfate. The magnesium sulfate was removed, and the ethyl acetate was evaporated under reduced pressure. The residue was purified by PTLC, and recrystallized from hexane. The obtained compound was dissolved in ethanol, 5% palladium carbon (150 mg) was added thereto, and the mixture was stirred overnight under hydrogen. The next day, the palladium carbon was removed, and the residue was concentrated, and the residue was recrystallized from ether to give the objective compound (0.172 g, 0.51 mmol, 17.1%).
The compounds of Examples 100 and 119 shown in the following Table 1 were also synthesized in the same manner as in Example 103.
The compound (0.191 g, 0.5 mmol) obtained in Example 107 was dissolved in THF (2 ml), and the pH of the solution was adjusted to 10 with 5% lithium hydroxide. The mixture was stirred for 30 minutes, and the resultant product was confirmed by TLC. The mixture was mildly acidified with 5% aqueous citric acid solution, and extracted with ethyl acetate. The extract was dried over magnesium sulfate, and the magnesium sulfate was removed. The residue was concentrated under reduced pressure, and the residue was recrystallized from hexane to give the objective compound (0.162 g, 0.44 mmol, 88.0%).
The compounds of Examples 110, 113, 115, 117, 121, and 123 shown in the following Table 1 were also synthesized in the same manner as in Example 108.
1H NMR ((CD3)2SO) δ = 3.66 (s, 3H), 4.87-4.88 (m, 1H), 6.35 (d, J = 8.2 Hz, 2H), 6.96-6.99 (m, 1H), 7.06-7.09 (m, 1H), 7.16-7.20 (m, 2H), 7.35 (d, J = 8.0 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 9.31 (d, J = 6.8 Hz, 1H), 11.0 (bs, 1H) MS (ESI) m/z: 353.1 (M −1) Yield: 34%
1H NMR ((CD3)2SO) δ = 3.66 (s, 3H), 4.86-4.88 (m, 1H), 6.35 (d, J = 8.2 Hz, 2H), 6.96-6.99 (m, 1H), 7.06-7.09 (m, 1H), 7.16-7.20 (m, 2H), 7.35 (d, J = 8.0 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 9.31 (d, J = 6.8 Hz, 1H), 11.0 (bs, 1H) MS (ESI) m/z: 353.1 (M − 1) Yield: 31%
1H NMR ((CD3)2SO) δ = 3.28-3.38 (m, 2H), 4.79 (bs, 1H), 6.34 (d, J = 8.2 Hz, 2H), 6.93-6.97 (m, 1H), 7.04-7.08 (m, 1H), 7.13-7.18 (m, 2H), 7.34 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 9.31 (bs, 1H), 10.9 (s, 1H) MS (ESI) m/z: 338.8 (M − 1) Yield: 94%
1H NMR ((CD3)2SO) δ = 3.26-3.34 (m, 2H), 4.79 (bs, 1H), 6.34 (d, J = 8.2 Hz, 2H), 6.93-6.97 (m, 1H), 7.04-7.08 (m, 1H), 7.13-7.18 (m, 2H), 7.34 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 9.31 (bs, 1H), 10.9 (s, 1H) MS (ESI) m/z: 338.8 (M − 1) Yield: 89%
1H NMR ((CD3)2SO) δ = 2.68-2.80 (m, 2H), 3.54 (s, 1H), 3.92 (s, 1H), 4.45 (s, 1H), 4.83 (s, 1H), 6.35-6.37 (m, 2H), 6.95-7.05 (m, 3H), 7.32-7.35 (m, 2H), 9.46 (s, 2H), 10.92 (s, 1H) MS (ESI) m/z: 306.9 (M − 1) Yield: 35%
1H NMR (CD3OD) δ = 3.01 (t, J = 7.4 Hz, 2H), 3.67 (t, J = 7.4 Hz, 2H), 6.68-6.71 (m, 1H), 6.87-6.91 (m, 2H), 7.02-7.05 (m, 2H), 7.17-7.20 (m, 1H), 7.36-7.40 (m, 1H), 7.71-7.74 (m, 1H) MS (ESI) m/z: 294.9 (M − 1) Yield: 78%
1H NMR ((CD3)2SO) δ = 2.93 (t, J = 7.7 Hz, 2H), 3.79-3.83 (m, 2H), 6.59-6.61 (m, 1H), 6.90 (d J = 2.3 Hz, 1H), 7.07-7.13 (m, 2H), 7.82-7.88 (m, 4H), 8.64 (s, 1H), 10.51 (s, 1H) MS (ESI) m/z: 361.0 (M + Na) Yield: 85%
1H NMR (CD3OD) δ = 3.04 (t, J = 7.3 Hz, 2H), 3.68 (t, J = 7.3 Hz, 2H), 6.67-6.70 (m, 1H), 7.00-7.20 (m, 3H), 7.62-7.84 (m, 3H), 7.85 (d, J = 6.7 Hz, 1H) Yield: 21%
1H NMR ((CD3)2SO) δ = 2.93 (t, J = 7.7 Hz, 2H), 3.79-3.83 (m, 2H), 6.59-6.61 (m, 1H), 6.90 (d J = 2.3 Hz, 1H), 7.09-7.15 (m, 2H), 7.41-7.43 (m, 1H), 7.48-7.55 (m, 2H), 7.74-7.77 (m, 1H), 8.53 (bs, 1H), 10.49 (s, 1H) MS (ESI) m/z: 323.0 (M − 1) Yield: quant.
1H NMR (CD3OD) δ = 3.30-3.34 (m, 2H), 3.71 (s, 3H), 4.93-4.96 (m, 1H), 6.35 (d, J = 8.3 Hz, 1H), 6.67-6.70 (m, 1H), 6.91-6.92 (m, 1H), 7.05 (s, 1H), 7.12-7.19 (m, 2H) MS (ESI) m/z: 368.9 (M − 1) Yield: 58%
1H NMR (CD3OD) δ = 3.33-3.38 (m, 1H), 3.67 (s, 3H), 3.73 (s, 3H), 4.94-4.97 (m, 1H), 6.36 (d, J = 8.2 Hz, 2H), 7.00-7.04 (m, 2H), 7.12-7.17 (m, 2H), 7.30 (d, J = 8.1 Hz, 1H), 7.49 (d, J = 8.1 Hz, 1H) MS (ESI) m/z: 366.9 (M − 1) Yield: 66%
1H NMR (CD3OD) δ = 3.32-3.35 (m, 2H), 6.34 (d, J = 8.2 Hz, 2H), 6.67-6.69 (m, 1H), 7.01-7.18 (m, 4H) Yield: 96%
1H NMR (CD3OD) δ = 3.32-3.35 (m, 2H), 3.63 (s, 3H), 4.93-5.03 (m, 1H), 6.34 (d, J = 8.2 Hz, 2H), 6.95-7.01 (m, 2H), 7.10-7.25 (m, 2H), 7.56 (d, J = 8.1 Hz, 1H), 7.24 (d, J = 8.2 Hz, 1H) MS (ESI) m/z: 352.5 (M − 1) Yield: 95%
1H NMR (CD3OD) δ = 3.11 (d, J = 6.8 Hz, 2H), 3.67 (d, J = 4.6 Hz, 2H), 4.44-4.47 (m, 1H), 6.37 (d, J = 8.2 Hz, 2H), 7.01-7.04 (m, 1H), 7.08-7.15 (m, 3H), 7.34 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H) MS (ESI) m/z: 324.8 (M − 1) Yield: 49%
1H NMR (CD3OD) δ = 3.33-3.38 (m, 2H), 5.00-5.03 (m, 3H), 6.36 (d, J = 8.2 Hz, 2H), 6.97-7.00 (m, 2H), 7.08-7.14 (m, 4H), 7.24 (s, 3H), 7.31-7.35 (m, 2H), 7.50 (d, J = 8.0 Hz, 1H) MS (ESI) m/z: 429.1 (M − 1) Yield: 52%
1H NMR (CD3OD) δ = 1.68 (s, 3H), 3.45 (d, J = 14.4 Hz, 1H), 3.51 (d, J = 14.4 Hz, 1H), 3.68 (s, 3H), 6.36 (d, J = 8.2 Hz, 2H), 6.95-6.99 (m, 1H), 7.05-7.09 (m, 2H), 7.12-7.17 (m, 1H), 7.33 (d, J = 8.0 Hz, 1H), 7.49 (d, J = 8.0 Hz, 1H) MS (ESI) m/z: 367.1 (M − 1) Yield: 32%
1H NMR (CD3OD) δ = 3.37 (d, J = 6.0 Hz, 2H), 3.69 (s, 3H), 4.96-4.99 (m, 1H), 6.35 (d, J = 8.2 Hz, 2H), 6.75-6.81 (m, 1H), 7.03-7.06 (m, 1H), 7.09 (s, 1H), 7.13-7.17 (m, 1H), 7.43-7.46 (m, 1H) MS (ESI) m/z: 371.0 (M − 1) Yield: 63%
1H NMR (CD3OD) δ = 1.73 (s, 3H), 3.49 (d, J = 14.5 Hz, 1H), 3.63 (d, J = 14.5 Hz, 1H), 6.34 (d, J = 8.2 Hz, 2H), 6.92-6.96 (m, 1H), 7.03-7.07 (m, 2H), 7.11-7.15 (m, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.56 (d, J = 8.0 Hz, 1H) MS (ESI) m/z: 352.8 (M − 1) Yield: 83%
1H NMR (CD3OD) δ = 3.40-3.42 (m, 2H), 4.94-4.97 (m, 1H), 6.34 (d, J = 8.2 Hz, 2H), 6.73-6.78 (m, 1H), 7.01-7.04 (m, 1H), 7.11-7.16 (m, 2H), 7.50-7.53 (m, 1H) MS (ESI) m/z: 356.8 (M − 1) Yield: 86%
1H NMR (CD3OD) δ = 1.40 (s, 9H), 3.32-3.33 (m, 1H), 4.59-4.67 (m, 1H), 6.42 (d, J = 8.2 Hz, 2H), 6.90-6.95 (m, 1H), 7.02-7.13 (m, 2H), 7.11 (s, 1H), 7.35 (d, J = 8.1 Hz, 1H), 7.65-7.70 (m, 1H) Yield: 86%
1H NMR (CD3OD) δ = 2.99-3.09 (m, 1H), 3.38-3.46 (m, 1H), 3.92- 3.93 (m, 1H), 6.41 (d, J = 8.2 Hz, 2H), 6.88-6.92 (m, 1H), 7.04-7.06 (m, 1H), 7.10-7.12 (m, 1H), 7.19 (s, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H) MS (ESI) m/z: 309.8 (M − 1) Yield: 23%
1H NMR ((CD3)2SO) δ = 2.19 (s, 6H), 2.86 (t, J = 7.6 Hz, 2H), 3.49-3.54 (m, 2H), 6.59-6.62 (m, 1H), 6.89 (d J = 2.3 Hz, 1H), 7.03 (d J = 7.5 Hz, 2H), 7.10-7.17 (m, 3H), 8.37-8.40 (m, 1H), 8.61 (s, 1H), 10.50 (s, 1H) MS (ESI) m/z: 307.0 (M − 1) Yield: 81%
1H NMR ((CD3)2SO) δ = 2.81 (t, J = 7.6 Hz, 2H), 3.40-3.44 (m, 2H), 3.74 (s, 6H), 6.60-6.63 (m, 1H), 6.66 (d J = 8.4 Hz, 2H), 6.88 (d J = 2.3 Hz, 1H), 7.10-7.16 (m, 2H), 7.26-7.30 (m, 1H), 8.09-8.15 (m, 1H), 8.62 (s, 1H), 10.51 (s, 1H) MS (ESI) m/z: 338.9 (M − 1) Yield: 43%
1H NMR ((CD3)2SO) δ = 2.82 (t, J = Hz, 2H), 3.46-3.51 (m, 2H), 6.60-6.62 (m, 1H), 6.87 (d J = 1.9 Hz, 1H), 7.10 (d J = 1.9 Hz, 1H), 7.13-7.15 (m, 1H), 7.84-7.88 (m, 1H), 8.13 (d J = 8.0 Hz, 2H), 8.63 (s, 1H), 8.87-8.90 (m, 1H), 10.53 (s, 1H) MS (ESI) m/z: 415.0 (M − 1) Yield: 20%
1H NMR ((CD3)2SO) δ = 2.87 (t, J = 7.5 Hz, 2H), 3.48-3.53 (m, 2H), 6.60-6.62 (m, 1H), 6.88 (d J = 2.3 Hz, 1H), 7.11-7.15 (m, 2H), 7.40-7.44 (m, 1H), 7.49-7.51 (2H), 8.62 (s, 1H), 8.79-8.82 (1H), 10.51 (s, 1H) MS (ESI) m/z: 346.8 (M − 1) Yield: 65%
1H NMR ((CD3)2SO) δ = 2.86 (t, J = 7.2 Hz, 2H), 3.55-3.58 (m, 2H), 5.80 (s, 2H), 6.60-6.63 (m, 1H), 6.91 (d J = 2.3 Hz, 1H), 7.08 (d J = 2.3 Hz, 1H), 7.14 (d J = 8.6 Hz, 1H), 8.61 (s, 1H), 8.69-8.76 (m, 1H), 9.87 (s, 1H), 10.51 (s, 1H) MS (ESI) m/z: 326.9 (M − 1) Yield: 35%
1H NMR ((CD3)2SO) δ = 2.15 (s, 3H), 2.89 (t, J = 7.2 Hz, 2H), 3.59-3.64 (m, 2H), 6.18 (s, 2H), 6.61-6.64 (m, 1H), 6.92 (d J = 2.3 Hz, 1H), 7.09 (d J = 2.3 Hz, 1H), 7.15 (d J = 8.6 Hz, 1H), 8.60 (bs, 1H), 8.93 (s, 1H), 10.53 (s, 1H) MS (ESI) m/z: 325.0 (M − 1) Yield: 29%
1H NMR (DMSO, 400 MHz) δ = 10.15 (1H, s), 9.14 (1H, s), 8.93 (1H, t, J = 5.64 Hz), 8.61 (1H, s), 7.30 (1H, d, J = 8.20 Hz), 7.14 (1H, d, J = 8.64 Hz), 7.08 (1H, s), 6.92 (1H, s), 6.90 (1H, d, J = 3.80 Hz), 6.69 (1H, t, J = 7.96 Hz), 6.61 (1H, d, J = 8.56 Hz), 3.52 (2H, dd, J = 6.24, 13.24 Hz), 2.87 (2H, t, J = 8.00 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310.9 Yield 23.0%
1H NMR (DMSO, 400 MHz) δ = 10.50 (1H, s), 10.06 (1H, s), 8.68 (1H, t, J = 5.48 Hz), 8.62 (1H, s), 7.69 (1H, d, J = 8.8 Hz), 7.14 (1H, d, J = 8.64 Hz), 7.07 (1H, s), 6.88 (1H, s), 6.61 (1H, d, J = 8.6 Hz), 6.30 (1H, d, J = 8.68 Hz), 6.23 (1H, s), 3.51 (2H, dd, J = 6.48, 13.08 Hz), 2.85 (2H, t, J = 7.96 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310. Yield 34.3%
1H NMR (DMSO, 400 MHz) δ = 10.51 (1H, s), 9.00 (1H, s), 8.82 (1H, s), 8.62 (1H, s), 7.24 (1H, s), 7.13 (1H, d, J = 8.60 Hz), 7.08 (1H, s), 6.86 (1H, d, J = 8.76), 6.74 (1H, d, J = 8.76 Hz), 6.61 (1H, d, J = 8.60 Hz), 3.53 (2H, dd, J = 6.68, 12.88 Hz), 2.86 (2H, t, J = 7.80 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310.8 Yield 30.1%
1H NMR (DMSO, 400 MHz) δ = 10.54 (1H, s), 9.01 (1H, s), 8.62 (1H, s), 7.16 (1H, s), 7.14 (1H, d, J = 8.56 Hz), 7.10 (1H, s), 6.90 (1H, s), 6.62 (1H, d, J = 8.56 Hz), 6.33 (2H, d, J = 8.20 Hz), 3.61 (2H, dd, J = 6.92, 12.68 Hz), 2.88 (2H, t, J = 7.56 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310.9 Yield 14.9%
1H NMR (DMSO, 400 MHz) δ = 10.47 (1H, s), 9.43 (1H, s), 9.09 (1H, s), 8.61 (1H, s), 8.25 (1H, t, J = 5.56), 7.29 (1H, s), 7.20 (1H, d, J = 8.24 Hz), 7.13 (1H, d, J = 8.60 Hz), 7.05 (1H, s), 6.88 (1H, s), 6.76 (1H, d, J = 8.24 Hz), 6.60 (1H, d, J = 8.56 Hz), 3.44 (2H, dd, J = 6.44, 13.20 Hz), 2.81 (2H, t, J = 8.04 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310.9
1H NMR (DMSO, 400 MHz) δ = 10.48 (1H, s), 9.44 (1H, s), 8.60 (1H, s), 8.35 (1H, t, J = 5.40 Hz), 7.13 (1H, d, J = 8.60 Hz), 7.05 (1H, s), 6.87 (1H, s), 6.68 (2H, s), 6.60 (1H, d, J = 8.56 Hz), 6.35 (1H, s), 3.43 (2H, dd, J = 6.60, 14.76 Hz), 2.81 (2H, t, J = 8.04 Hz). ESI-MS [M + H]+ = 313.1 [M − H]− = 310.8 Yield 60.5%
1H NMR (DMSO, 400 MHz) δ = 10.80 (1H, s), 7.94 (1H, t, J = 5.68, 11.36 Hz), 7.52 (1H, d, J = 7.80 Hz), 7.35-6.96 (8H, m), 3.19-3.3 (1H, m), 3.09-3.05 (2H, m), 2.95-2.90 (1H, m). ESI-MS [M + H]+ = 307.6 [M − H]− = 305.9 Yield 48.7%
1H NMR (MeOD, 400 MHz) δ = 7.61 (1H, d, J = 7.92 Hz), 7.34 (1H, d, J = 8.12 Hz), 7.22 (1H, d, J = 8.12 Hz), 7.13-7.07 (2H, m), 7.01-6.92 (2H, m), 6.71 (1H, t, J = 7.96 Hz), 3.51-3.37 (2H, m). ESI-MS [M + H]+ = 341.1 [M − H]− = 338.8 Yield 5.8%
1H NMR (DMSO, 400 MHz) δ = 10.47 (1H, s), 8.61 (1H, s), 7.91 (1H, t, J = 5.68 Hz), 7.29-7.18 (4H, m), 7.13 (1H, d, J = 8.60 Hz), 6.99 (1H, s), 6.83 (1H, s), 6.60 (1H, d, J = 8.6 Hz), 4.36 (1H, bs), 3.48-3.42 (2H, m), 2.93 (1H, dd, J = 5.12, 13.36 Hz), 2.69-2.64 (2H, m). ESI-MS [M + H]+ = 324.2 [M − H]− = 322.0 Yield 43.4%
1H NMR (DMSO, 400 MHz) δ = 10.84 (1H, s), 9.03 (1H, bs), 8.90 (1H, bs), 7.56 (1H, d, J = 7.88 Hz), 7.33-7.30 (2H, m), 7.14 (1H, s), 7.05 (1H, t, J = 7.04, 14.08 Hz), 6.97, 6.96, 6.94 (1H, t, J = 7.04, 14.00 Hz), 6.82 (1H, d, J = 5.76 Hz), 6.73 (1H, d, J = 8.76), 4.68 (1H, bs), 3.47-3.24 (2H, m). ESI-MS [M − H]− = 339.0 Yield 34.8%
1H NMR (DMSO, 400 MHz) δ = 10.83 (1H, s), 8.99 (1H, s), 8.83 (1H, s), 7.58 (1H, d, J = 7.8 Hz), 7.36 (1H, d, J = 8.04 Hz), 7.24 (1H, s), 7.19 (1H, s), 7.08 (1H, t, J = 7.00, 14.04 Hz), 6.99 (1H, t, J = 7.92, 14.84 Hz), 6.87 (1H, dd, J = 2.92, 8.80 Hz), 6.75 (1H, d, J = 8.80 Hz), 3.57 (2H, dd, J = 6.92, 13.04 Hz), 2.97 (2H, t, J = 7.88 Hz). ESI-MS [M + H]+ = 297.0 [M − H]− = 294.9 Yield 68.6%
1H NMR (DMSO, 400 MHz) δ = 10.89 (1H, s), 8.23 (1H, d, J = 7.60 Hz), 7.59 (1H, d, J = 7.84 Hz), 7.36 (1H, d, J = 8.08 Hz), 7.25 (1H, d, J = 2.20 Hz), 7.07 (1H, t, J = 7.00 Hz), 6.98 (1H, t, J = 7.52), 6.91-6.85 (2H, m), 6.80-6.76 (1H, m), 3.65, 3.64, 3.63, 3.62 (1H, dd, J = 3.88, 8.88 Hz), 3.27 (1H, d, J = 14.32 Hz), 2.85 (1H, dd, J = 8.88, 14.36 Hz). ESI-MS [M + H]+ = 296.1 [M − H]− = 294.1 Yield 72.8%
1H NMR (DMSO, 400 MHz) δ = 10.49 (1H, s), 8.59 (2H, s), 7.87 (2H, d, J = 6.96 Hz), 7.53-7.45 (3H, m), 7.14 (1H, d, J = 8.56 Hz), 7.07 (1H, s), 6.90 (1H, s), 6.61 (1H, d, J = 8.60 Hz), 3.52 (2H, dd, J = 6.56, 13.28 Hz), 2.86 (2H, t, J = 7.84, 14.84 Hz). ESI-MS [M + H]+ = 281.0 [M − H]− = 278.8 Yield 84.6%
1H NMR (DMSO, 400 MHz) δ = 10.08 (1H, s), 9.17 (1H, s), 7.89 (1H, t, J = 5.84, 11.64 Hz), 7.55 (1H, d, J = 7.84), 7.34 (1H, d, J = 8.04 Hz), 7.12 (1H, s), 7.08 (1H, t, J = 6.96, J = 13.92 Hz), 7.00-6.96 (3H, m), 6.67 (2H, d, J = 4.48 Hz), 3.33-3.27 (2H, m), 2.83-2.75 (3H, m), 1.57 (2H, s). ESI-MS [M + H]+ = 324.2 [M − H]− = 321.8 Yield 82.9%
1H NMR (DMSO, 400 MHz) δ = 10.48 (NH), 8.07 (NH, amide), 7.24 (d, J = 15.6 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H), 7.05 (s, 1H), 6.94 (d, J = 2.0 Hz, 1H), 6.84 (d, J = 2.0 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 6.74 (d, J = 8.1 Hz, 1H), 6.59 (dd, J = 8.6, 2.3 Hz, 1H), 6.33 (d, J = 15.6 Hz, 1H), 3.44 (m, 2H), 2.78 (m, 2H). ESI-MS: [M + H]+ = 339.1, [M + Na]+ = 361.0, [M − H]− = 337.2, [2M − H]− = 675.2. Yield: 54.0%
1H NMR (DMSO, 400 MHz) δ = 10.48 (NH), 8.07 (NH, amide), 7.24 (d, J = 15.6 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H), 7.05 (s, 1H), 6.94 (d, J = 2.0 Hz, 1H), 6.84 (d, J = 2.0 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 6.74 (d, J = 8.1 Hz, 1H), 6.59 (dd, J = 8.6, 2.3 Hz, 1H), 6.33 (d, J = 15.6 Hz, 1H), 3.44 (m, 2H), 2.78 (m, 2H). ESI-MS: [M + H]+ = 339.1, [M + Na]+ = 361.0, [M − H]− = 337.2, [2M − H]− = 675.2. Yield: 54.0%
1H NMR (DMSO, 400 MHz) δ = 10.47 (NH), 8.74 (OH) 8.57 (OH), 7.97 (NH), 7.11 (d, J = 8.6 Hz, 1H), 7.00 (d, J = 2.2 Hz, 1H), 6.82 (m, 2H), 6.68 (d, J = 8 Hz, 1H), 6.62 (dd, J = 8, 1.8 Hz, 1H), 6.58 (dd, J = 8.6, 2.2 Hz, 1H), 3.73 (s, 3H), 3.28 (m, 4H), 2.71 (t, J = 7.4 Hz, 2H). ESI-MS: [M + H]+ = 341.2, [M + Na]+ = 363.1, [2M − H]− = 339.2, [2M − H]− = 679.3. Yield: 56.0%
1H-NMR (CDCl3) δ ppm: 0.66 (3H, d, J = 6.9 Hz), 0.86 (6H, d, J = 7.2 Hz), 1.24 (1H, m), 1.26 (1H, m), 1.48 (1H, m), 1.61-1.70 (4H, m), 1.86 (1H, dt, J = 3.5, 11.6), 2.98 (2H, m), 3.61 (2H, m), 5.43 (1H, m), 7.02 (1H, 2.4 Hz), 7.13 (1H, m), 7.22 (1H, m), 7.39 (1H, m), 7.63 (1H, m), 8.06 (1H, bs) ESI-MS: 327.3 (M + H)+, 325.3 (M − H)−
1H-NMR (CDCl3) δ ppm: 0.61 (3H, d, J = 6.9 Hz), 0.82 (3H, d, J = 6.9 Hz), 0.83 (3H, d, J = 6.5 Hz), 0.91 (2H, m), 1.12 (1H, m), 1.26 (1H, m), 1.46 (1H, tt, J = 0.5, 11.1 Hz), 1.56-1.70 (3H, m), 1.73 (1H, m), 1.95 (1H, dt, J = 3.6, 11.8 Hz), 3.31 (2H, m), 3.69 (3H, s), 5.02 (1H, m), 7.00 (1H, d, J = 2.4 Hz), 7.11 (1H, m), 7.20 (1H, m), 7.36 (1H, m), 7.55 (1H, m), 8.06 (1H, bs)
1H-NMR (CDCl3) δ ppm: 0.60 (3H, d, J = 6.9 Hz), 0.81 (3H, d, J = 7.0 Hz), 0.82 (3H, d, J = 6.6 Hz), 0.92 (2H, m), 1.11 (1H, m), 1.26 (1H, m), 1.46 (1H, tt, J = 1.4, 11.0 Hz), 1.57-1.71 (4H, m) , 1.92 (1H, dt, J = 3.2, 8.4 Hz), 3.24 (1H, dd, J = 5.8, 14.9 Hz), 3.35 (1H, dd, J = 5.8, 14.9 Hz), 5.07 (1H, m), 5.08 (1H, d, J = 12.2 Hz), 5.13 (1H, d, J = 12.2 Hz), 5.87 (1H, d, J = 8.0 Hz), 6.80 (1H, d, J = 2.4 Hz), 7.10 (m, 1H), 7.19 (1H, m), 7.24 (1H, m), 7.32-7.36 (4H, m), 7.55 (1H, m), 7.97 (1H, bs) ESI-MS: 385.2 (M + H)+, 383.2 (M − H)−
1H-NMR (CDCl3) δ ppm: 0.49 (3H, d, J = 6.8 Hz), 0.76 (3H, d, J = 6.8 Hz), 0.82 (3H, d, J = 6.5 Hz), 0.88 (2H, m), 1.07 (1H, m), 1.22 (1H, m), 1.40 (2H, m), 1.57-1.69 (3H, m), 1.90 (1H, dt, J = 3.3, 11.8 Hz), 3.36 (1H, dd, J = 6.2 Hz), 4.92 (1H, dd, J = 6.2, 13.2 Hz), 5.93 (1H, d, J = 7.1 Hz), 7.08 (1H, d, J = 2.4 Hz), 7.13 (1H, m), 7.21 (1H, m), 7.37 (1H, d, J = 8.1 Hz), 7.60 (1H, d, J = 7.8 Hz), 8.22 (1H, s) ESI-MS: 461.3 (M + H)+, 459.3 (M − H)−
1H-NMR (CDCl3) δ ppm: 0.64 (3H, d, J = 6.8 Hz), 0.84 (3H, d, J = 6.8 Hz), 0.86 (3H, d, J = 6.4 Hz), 0.88 (2H, m), 0.93 (2H, m), 1.17 (1H, m), 1.28 (1H, m), 1.58-1.73 (4H, m), 1.92 (1H, t, J = 5.8 Hz), 1.99 (1H, dd, J = 3.4, 11.6 Hz), 3.11 (1H, dd, J = 9.1, 14.5 Hz), 3.20 (1H, m), 3.28 (1H, m), 3.35 (1H, dd, J = 5.4, 14.5 Hz), 4.71 (1H, m), 5.96 (1H, m), 6.18 (1H, d, J = 76. Hz), 7.12 (1H, d, J = 2.32 Hz), 7.15 (1H, m), 7.23 (1H, m), 7.38 (1H, m), 7.74 (1H, d, J = 8.4 Hz), 8.09 (1H, m) ESI-MS: 414.2 (M + H)+, 412.2 (M − H)−
1H-NMR (CDCl3) δ ppm: 1.43 (9H, s), 2.06 (1H, m), 3.13-3.36 (4H, m), 3.46-3.50 (2H, m), 4.38 (1H, m), 5.17 (1H, m), 6.00 (1H, bs), 7.10 (1H, d, J = 2.4 Hz), 7.15 (1H, m), 7.22 (1H, m), 7.38 (1H, m), 7.67 (1H, dd, J = 7.7 Hz), 8.13 (1H, bs) ESI-MS: 369.9 (M + Na)+, 346.0 (M − H)−
1H-NMR (CDCl3) δ ppm: 1.43 (9H, s), 2.20 (1H, m), 3.13-3.35 (4H, m), 3.43-3.47 (2H, m), 4.39 (1H, m), 5.21 (1H, m), 6.06 (1H, bs), 7.08 (1H, d, J = 2.2 Hz), 7.14 (1H, m), 7.21 (1H, m), 7.37 (1H, m), 7.66 (1H, dd, J = 7.8 Hz), 8.24 (1H, bs) ESI-MS: 369.5 (M + Na)+, 346.0 (M − H)−
1H-NMR (CDCl3) δ ppm: 1.95 (1H, bs), 3.12-3.25 (3H, m), 3.34- 3.46 (3H, m), 4.46 (1H, m), 5.11 (2H, s), 5.49 (1H, m), 5.95 (1H, m), 7.06 (1H, d, J = 1.6 Hz), 7.13 (1H, m), 7.21 (1H, m), 7.32-7.37 (6H, m), 7.66 (1H, m), 8.13 (1H, bs) ESI-MS: 404.1 (M + Na)+, 379.9 (M − H)−
1H-NMR (CDCl3) δ ppm: 1.43 (9H, s), 2.96 (2H, t, J = 6.8 Hz), 3.46 (2H, m), 4.60 (1H, m), 7.04 (1H, d, J = 2.0 Hz), 7.12 (1H, m), 7.20 (1H, m), 7.38 (1H, m), 7.61 (1H, d, J = 7.6 Hz), 8.00 (1H, bs) ESI-MS: 283.0 (M + Na)+, 258.9 (M − H)−
1H-NMR (CDCl3) δ ppm: 2.99 (1H, t, J = 6.7 Hz), 3.55 (2H, m), 4.81 (1H, m), 5.10 (2H, s), 7.01 (1H, d, J = 1.4 Hz), 7.11 (1H, m), 7.20 (1H, m), 7.29-7.38 (5H, m), 7.60 (1H, d, J = 7.6 Hz), 7.99 (1H, bs) ESI-MS: 316.7 (M + Na)+, 293.0 (M − H)−
1H-NMR (CDCl3) δ ppm: 0.60 (3H, d, J = 6.8 Hz), 0.80 (3H, d, J = 6.8 Hz), 0.84 (3H, d, J = 6.4 Hz), 0.91 (2H, m), 1.10 (1H, m), 1.26 (1H, m), 1.43-1.54 (3H, m), 1.60-1.69 (2H, m), 1.91 (1H, dt, J = 3.6, 11.6 Hz), 2.88 (1H, dd, J = 6.8, 14.0 Hz), 2.96-3.09 (2H, m), 3.26 (1H, dd, J = 5.0, 14.3 Hz), 4.65 (1H, m), 4.75 (1H, dd, J = 2.6, 5.7 Hz), 6.22 (1H, d, J = 7.3 Hz), 6.38 (1H, d, J = 7.0 Hz), 6.89 (1H, d, J = 6.2 Hz), 6.95 (s, 1H), 7.10-7.20 (6H, m), 7.30 (1H, d, J = 8.0 Hz), 7.70 (1H, d, J = 7.5 Hz), 8.26 (1H, s) ESI-MS: 540.3 (M + Na)+, 516.0 (M − H)−
1H-NMR (CDCl3) δ ppm: 0.90 (9H, s), 1.98 (m, 2H), 3.37 (2H, m), 4.88 (1H, dd, J = 6.2, 12.8 Hz), 5.92 (1H, d, J = 7.0 Hz), 7.06 (1H, d, J = 1.6 Hz), 7.13 (1H, m), 7.21 (1H, m), 7.36 (1H, d, J = 8.1 Hz), 7.59 (1H, d, J = 7.9 Hz), 8.23 (1H, bs) ESI-MS: 325.2 (M + Na)+, 301.2 (M − H)−
1H-NMR (CDCl3) δ ppm: 2.42 (2H, m), 2.89 (2H, m), 3.30 (2H, d, J = 5.4), 4.88 (1H, m), 5.94 (1H, d, J = 7.4 Hz), 6.86 (1H, d, J = 2.2 Hz), 7.07-7.12 (3H, m), 7.17- 7.21 (2H, m), 7.23 (1H, m), 7.25 (1H, m), 7.34 (1H, d, J = 8.1 Hz), 7.48 (1H, d, J = 7.8 Hz), 8.10 (1H, s) ESI-MS: 359.1 (M + Na)+, 334.9 (M − H)−
1H-NMR (CDCl3) δ ppm: 2.38 (2H, m), 2.82 (2H, m), 3.29 (2H, d, J = 5.7), 3.76 (3H, s), 4.87 (1H, m), 5.93 (1H, d, J = 7.2 Hz), 6.78 (2H, m), 6.83 (1H, m), 7.01 (2H, m), 7.10 (1H, m), 7.19 (1H, m), 7.34 (1H, d, J = 8.1 Hz), 7.48 (1H, d, J = 8.0), 8.14 (1H, s) ESI-MS: 389.2 (M + Na)+, 365.0 (M − H)−
1H-NMR (CDCl3) δ ppm: 2.36 (2H, m), 2.79 (2H, m), 3.31 (2H, d, J = 5.4), 4.87 (1H, m), 5.89 (2H, m), 5.93 (1H, d, J = 7.4 Hz), 6.54 (1H, m), 6.60 (1H, d, J = 1.6 Hz), 6.67 (1H, d, J = 6.8 Hz), 6.93 (1H, s), 7.13 (1H, m), 7.20 (1H, m), 7.35 (1H, d, J = 8.1 Hz), 7.50 (1H, d, J = 7.8 Hz), 8.16 (1H, m) ESI-MS: 419.0 (M + Na)+, 378.9 (M − H)−
1H-NMR (CDCl3) δ ppm: 2.43 (2H, m), 2.81 (2H, m), 3.26 (2H, m), 3.85 (3H, s), 5.00 (1H, m), 5.09 (2H, s), 5.58 (1H, s), 5.91 (1H, d, J = 8.0 Hz), 6.59-6.74 (2H, m), 7.06 (1H, m), 7.18 (1H, m), 7.25 (2H, m), 7.36 (4H, m), 7.43 (1H, m), 7.96 (1H, bs) ESI-MS: 495.1 (M + Na)+, 471.1 (M − H)−
1H-NMR (CD3OD) δ ppm: 2.39 (2H, m), 2.68 (2H, m), 3.17 (2H, dd, J = 7.4, 14.7 Hz), 3.80 (3H, s), 3.84 (1H, m), 4.69 (1H, dd, J = 5.0, 7.4 Hz), 6.55 (1H, dd, J = 2.1, 8.2 Hz), 6.65 (1H, d, J = 2.1 Hz), 6.75 (1H, d, J = 8.2 Hz), 6.98-7.02 (2H, m), 7.08 (1H, m), 7.32 (1H, d, J = 8.1 Hz), 7.55 (1H, d, J = 8.1 Hz) ESI-MS: 405.1 (M + Na)+, 381.1 (M − H)−
1H-NMR (CD3Cl) δ ppm: 2.39 (2H, m), 2.78 (2H, m), 3.22 (2H, d, J = 5.4 Hz), 3.49 (1H, d, J = 3.8 Hz), 4.93 (1H, m), 5.06 (1H, s), 5.76 (1H, s), 6.01 (1H, d, J = 7.8 Hz), 6.48-6.53 (2H, m), 6.61 (1H, s), 6.65 (1H, d, J = 2.0 Hz), 6.73 (1H, d, J = 8.0 Hz), 7.06 (1H, m), 7.12 (1H, m), 7.22 (2H, m), 7.30-7.35 (3H, m), 7.40 (1H, d, J = 8.1 Hz), 8.03 (1H, bs) ESI-MS: 481.0 (M + Na)+, 457.0 (M − H)−
1H-NMR (CD3OD) δ ppm: 2.35 (2H, m), 2.55 (2H, m), 3.18 (1H, dd, J = 6.5, 14.2 Hz), 4.60 (1H, m), 4.64 (1H, dd, J = 4.9, 6.5 Hz), 6.46 (1H, m), 6.61-6.64 (2H, m), 6.97- 7.01 (2H, m), 6.97-7.01 (2H, m), 7.04-7.08 (1H, m), 7.30 (1H, m), 7.56 (1H, m) ESI-MS: 391.1 (M + Na)+, 367.1 (M − H)−
1H-NMR (DMSO) δ ppm: 2.40 (2H, t, J = 7.5 Hz), 2.74 (2H, t, J = 7.5 Hz), 2.98 (1H, dd, J = 8.3, 14.3 Hz), 3.15 (1H, dd, J = 4.4, 14.3 Hz), 3.71 (3H, s), 4.48 (1H, m), 7.02 (1H, m), 7.11-7.36 (6H, m), 7.37 (1H, d, J = 2.3 Hz), 7.37 (1H, d, J = 8.2 Hz), 7.54 (1H, d, J = 7.9 Hz), 8.15 (1H, d, J = 7.9 Hz) ESI-MS: 351.2 (M + H)+, 348.8 (M − H)−
1H-NMR (DMSO) δ ppm: 2.37 (2H, t, J = 7.0 Hz), 2.72 (2H, t, J = 7.0 Hz), 2.84 (1H, dd, J = 9.4, 13.7 Hz), 3.04 (1H, dd, J = 4.9, 13.7 Hz), 4.44 (1H, m), 7.15-7.32 (10H, m), 8.20 (1H, d, J = 4.3 Hz) ESI-MS: 298.2 (M + H)+, 295.9 (M − H)−
1H-NMR (DMSO) δ ppm: 2.37 (3H, s), 2.38 (2H, t, J = 8.0 Hz), 2.73 (2H, dt, J = 3.9, 8.0 Hz), 2.95 (1H, dd, J = 8.6, 14.3 Hz), 3.13 (1H, dd, J = 4.6, 8.6 Hz), 4.46 (1H, m), 6.89 (1H, dd, J = 1.3, 8.0 Hz), 7.04 (1H, d, J = 2.3 Hz), 7.15 (3H, m), 7.21 (3H, m), 7.31 (1H, s), 8.14 (1H, d, J = 7.9) ESI-MS: 351.2 (M + H)+, 348.9 (M − H)−
1H-NMR (DMSO) δ ppm: 2.39 (1H, t, J = 7.8 Hz), 2.75 (1H, t, J = 7.8 Hz), 2.88 (1H, dd, J = 8.8, 14.5 Hz), 3.05 (1H, dd, J = 5.2, 14.5 Hz), 4.43 (1H, m), 6.58 (1H, dd, J = 2.3, 8.6 Hz), 6.84 (1H, d, J = 2.3 Hz), 6.99 (1H, d, J = 2.3 Hz), 7.11-7.16 (4H, m), 7.23 (2H, m), 8.10 (1H, d, J = 7.7 Hz), 8.60 (1H, s), 10.5 (1H, s) ESI-MS: 350.9 (M − H)−
1H-NMR (DMSO) δ ppm: 1.37 (s, 3H), 2.50 (2H, t, J = 7.9 Hz), 2.93 (2H, t, J = 7.9 Hz), 3.15 (1H, d, J = 13.3 Hz), 3.37 (1H, d, 13.3 Hz), 7.01-7.03 (2H, m), 7.20-7.31 (8H, m), 7.78 (1H, s) ESI-MS: 334.2 (M + Na)+, 309.4 (M − H)−
1H-NMR (CD3OD) δ ppm: 2.47 (2H, t, J = 7.9 Hz), 2.81 (2H, t, J = 7.9 Hz), 3.12 (1H, dd, J = 8.1, 14.7 Hz), 3.29 (1H, dd, J = 5.0, 14.7 Hz), 4.73 (1H, dd, J = 5.0, 8.1 Hz), 6.80 (1H, m), 6.96 (1H, s), 7.03 (1H, m), 7.13-7.24 (4H, m), 7.49 (1H, dd, J = 5.2, 8.6 Hz), 10.4 (1H, s) ESI-MS: 355.2 (M + H)+, 353.1 (M − H)−
1H-NMR (CD3OD) δ ppm: 2.42 (3H, s), 2.46 (2H, t, J = 7.9 Hz), 2.80 (2H, m), 3.11 (1H, dd, J = 8.2, 14.7 Hz), 3.38 (1H, dd, J = 5.0, 14.7 Hz), 4.73 (1H, dd, J = 5.0, 8.2 Hz), 6.87 (1H, d, J = 8.1 Hz), 6.91 (1H, s), 7.11-7.23 (10H, m), 7.43 (1H, d, J = 8.1 Hz) ESI-MS: 373.1 (M + Na)+, 349.4 (M − H)−
1H-NMR (CD3OD) δ ppm: 2.56 (1H, m), 2.75 (1H, m), 3.10 (1H, dd, J = 11.3, 14.4 Hz), 3.35 (1H, dd, J = 5.0, 14.4 Hz), 5.17 (1H, dd, J = 5.0, 11.3 Hz), 7.00-7.31 (10H, m)
1H-NMR (CD3OD) δ ppm: 2.42 (2H, dt, J = 7.8, 1.4 Hz), 2.93 (2H, t, J = 7.8 Hz), 5.45 (1H, s), 7.17- 7.36 (10H, m) ESI-MS: 306.1 (M + Na)+, 281.8 (M − H)−
1H NMR (DMSO, 400 MHz) δ = 7.33-7.28 (4H, m), 7.26-7.21 (1H, m), 6.86 (1H, t, J = 8.12 Hz), 6.37 (2H, d, J = 8.12 Hz), 3.72 (1H, dd, J = 8.92 Hz), 3.18 (1H, dd, J = 13.6 Hz), 2.75 (1H, dd, J = 13.6 Hz). ESI-MS [M + H]+ = 273.2 [M − H]− = 270. 9 Yield 24.5%
1H NMR (DMSO, 400 MHz) δ = 7.31-7.16 (5H, m), 6.85 (1H, t, J = 8.12 Hz), 6.35 (2H, d, J = 8.12 Hz), 3.43 (1H, dd, J = 8.44 Hz), 2.75- 2.70 (2H, m), 2.08-2.05 (1H, m), 1.80-1.75 (1H, m). ESI-MS [M + H]+ = 286.9 [M − H]− = 284.9 Yield 18.1%
1H NMR (DMSO, 400 MHz) δ = 9.23 (1H, m), J = 6.88 Hz), 7.31- 7.15 (6H, m), 6.36 (2H, d, J = 8.20 Hz), 4.80 (dd, 1H, J = 6.36, 11.88 Hz), 3.25-3.11 (2H, m). ESI-MS [M + H]+ = 302.0 [M − H]− = 299.9 Yield 26.7%
1H NMR (DMSO, 400 MHz) δ = 7.30-7.26 (2H, m), 7.21-7.15 (4H, m), 6.38 (2H, d, J = 8.24 Hz), 4.49 (1H, t, J = 6.96, 12.32 Hz), 2.65 (1H, t, J = 7.80, 16.16 Hz), 2.21- 2.04 (1H, m). ESI-MS [M + H]+ = 316.2 [M − H]− = 314.3 Yield 34.7%
1H NMR (DMSO, 400 MHz) δ = 10.85 (1H, s), 9.03 (1H, s), 7.61 (1H, d, J = 7.88), 7.36 (1H, d, J = 8.04), 7.21-6.97 (4H, m), 6.35 (2H, d, J = 8.20), 3.68 (2H, dd, J = 7.04, 12.68 Hz), 3.00 (2H, t, J = 7.12, 14.28 Hz). ESI-MS [M + H]+ = 297.1 [M − H]− = 294.9 Yield 74.0%
1H NMR (DMSO, 400 MHz) δ = 9.76 (2H, s), 9.56 (1H, s), 7.52 (1H, d, J = 15.56 Hz), 7.21 (1H, s), 7.18 (1H, d, J = 8.12 Hz), 7.07- 6.99 (2H, m), 6.91 (1H, t, J = 8.08 Hz), 6.41 (2H, d, J = 8.12 Hz), 6.10 (2H, s). ESI-MS [M + H]+ = 300.2 [M − H]− = 298.1 Yield 12.5%
1H NMR (DMSO, 400 MHz) δ = 9.33 (3H, s), 6.90-6.85 (1H, m), 6.82 (1H, d, J = 7.88 Hz), 6.73 (1H, d, J = 7.92 Hz), 6.36 (2H, d, J = 8.08 Hz), 5.96 (2H, s), 2.84 (2H, dd, J = 7.04, 15.24 Hz), 2.70 (2H, dd, J = 8.28, 15.12 Hz). ESI-MS [M + H]+ = 297.1 [M − H]− = 294.9 Yield 48.7%
1H NMR (DMSO, 400 MHz) δ = 10.79 (1H, s), 9.39 (2H, bs), 7.58 (1H, d, J = 7.72 Hz), 7.33 (1H, d, J = 8.00 Hz), 7.17 (1H, s), 7.06 (1H, t, J = 7.04 Hz), 7.01 (1H, t, J = 7.08 Hz), 6.88 (1H, t, J = 8.12 Hz), 6.37 (2H, d, J = 8.12 Hz), 3.02 (2H, dd, J = 7.08, 15.28 Hz), 2.82 (2H, dd, J = 6.00, 13.00 Hz). ESI-MS [M + H]+ = 297.0 [M − H]− = 294.9 Yield 55.0%
1H NMR (DMSO, 400 MHz) δ = 9.73 (2H, s), 9.65 (1H, s), 9.25 (1H, s), 7.49 (1H, d, J = 15.6 Hz), 7.06 (2H, d, J = 9.24 Hz), 7.00-6.89 (3H, m), 6.41 (2H, d, J = 8.12 Hz), 3.82 (3H, s). ESI-MS [M + H]+ = 302.0 [M − H]− = 299.8 Yield 27.3%
1H NMR (DMSO, 400 MHz) δ = 9.36 (3H, s), 8.81 (1H, s), 6.87 (1H, t, J = 8.12 Hz), 6.82 (1H, d, J = 8.16 Hz), 6.68 (1H, s), 6.64 (1H, d, J = 8.16 Hz), 6.34 (2H, d, J = 8.12 Hz), 2.77 (2H, dd, J = 6.36 Hz), 2.68 (2H, dd, J = 4.92). ESI-MS [M + H]+ = 304.0 [M − H]− = 301.9 Yield 60.0%
1H NMR (DMSO, 400 MHz) δ = 9.76 (2H, s), 9.63 (1H, s), 7.59 (3H, d, J = 8.44 Hz), 7.04 (3H, d, J = 8.72 Hz) 6.91 (1H, t, J = 8.12 Hz), 6.40 (2H, d, J = 8.12 Hz), 3.81 (3H, s). ESI-MS [M + H]+ = 286.2 [M − H]− = 283.8 Yield 52.5%
1H NMR (DMSO, 400 MHz) δ = 9.34 (3H, s), 7.18 (2H, d, J = 8.60 Hz), 6.90-6.84 (3H, m), 6.37 (2H, d, J = 8.12 Hz), 3.71 (3H, s), 2.85 (2H, dd, J = 7.04, 15.40 Hz), 2.71 (2H, dd, J = 6.20 Hz) ESI-MS [M + H]+ = 288.0 [M − H]− = 286.1 Yield 31.6%
1H NMR (DMSO, 400 MHz) δ = 10.89 (1H, s), 9.11 (1H, d, J = 7.2 Hz), 8.08 (1H, s), 7.57-7.51 (2H, m), 7.35 (1H, d, J = 8.04 Hz), 7.18 (1H, s), 7.07 (1H, t, J = 7.00 Hz), 6.98 (1H, t, J = 7.00 Hz), 6.90 (1H, d, J = 8.8 Hz), 4.81 (1H, dd, J = 7.76, 13.04 Hz), 3.65 (3H, s), 3.33- 3.27 (2H, m). ESI-MS [M + H]+ = 417.0 [M − H]− = 414.9 Yield 73.0%
1H NMR (DMSO, 400 MHz) δ = 10.51 (1H, s), 9.01 (1H, s), 8.61 (1H, s), 8.07 (1H, s), 7.54 (1H, d, J = 8.80 Hz), 7.14 (1H, d, J = 8.60 Hz), 7.08 (1H, s), 6.90 (2H, d, J = 8.84 Hz), 6.60 (1H, d, J = 8.60 Hz), 3.55 (2H, dd, J = 6.84, 12.84 Hz), 2.89 (2H, dd, J = 7.64, 15.00 Hz). ESI-MS [M + H]+ = 374.9 [M − H]− = 373.0 Yield 49.3%
1H NMR (DMSO, 400 MHz) δ = 10.88 (1H, s), 9.04 (1H, d, J = 7.20 Hz), 7.92 (1H, d, J = 6.76 Hz), 7.53 (1H, d, J = 7.88 Hz), 7.39 (1H, t, J = 6.08 Hz), 7.33 (1H, d, J = 8.08 Hz), 7.19 (1H, s), 7.06 (1H, t, J = 6.96), 6.97 (1H, t, J = 6.96 Hz), 6.91 (1H, t, J = 7.08 Hz), 4.76 (1H, dd, J = 7.72, 13.44 Hz), 3.33- 3.28 (2H, m). ESI-MS [M + H]+ = 314.1 [M − H]− = 311.8 Yield 56.5%
1H NMR (DMSO, 400 MHz) δ = 10.84 (1H, s), 9.02 (1H, t, J =), 8.05 (1H, s), 7.59-7.53 (2H, m), 7.34 (1H, d), 7.19 (1H, s), 7.08 (1H, t, J =), 6.98 (1H, t, J =), 6.88 (1H, d, J =). ESI-MS [M + H]+ = 358.9 [M − H]− = 357.0 Yield 43.4%
1H NMR (DMSO, 400 MHz) δ = 9.07 (1H, d, J = 7.36 Hz), 8.04 (1H, s), 7.56 (1H, d, J = 8.76 Hz), 7.31- 7.20 (5H, m), 6.89 (1H, d, J = 8.76 Hz), 4.77 (1H, dd, J = 8.68, 12.96 Hz), 3.22-3.09 (2H, m). ESI-MS [M + H]+ = 379.9 [M − H]− = 377.9 Yield 66.0%
1H NMR (DMSO, 400 MHz) δ = 9.03 (1H, d, J = 7.44 Hz), 7.89 (1H, d, J = 6.68 Hz), 7.40 (1H, t, J = 7.20 Hz), 7.31-7.21 (5H, m), 6.91 (2H, d, J = 7.96 Hz), 4.72 (1H, q, J = 7.40 Hz), 3.16 (2H, t, J = 6.28 Hz). ESI-MS [M + H]+ = 353.2 [M − H]− = 351.1 Yield 57.5%
1H NMR (DMSO, 400 MHz) δ = 9.15 (1H, d, J = 7.28 Hz), 8.14 (1H, s), 7.59 (1H, d, J = 8.8 Hz), 7.29 (2H, t, J = 7.40 Hz), 7.23-7.18 (3H, m), 6.94 (1H, d, J = 8.80 Hz), 4.45 (1H, q, J = 7.12), 3.66 (3H, s), 2.74-2.66 (2H, m), 2.16-2.12 (2H, m). ESI-MS [M + H]+ = 394.1 [M − H]− = 392.0 Yield 26.6%
1H NMR (DMSO, 400 MHz) δ = 9.06 (1H, d, J = 7.28 Hz), 7.98 (1H, d, J = 8.20 Hz), 7.44 (1H, t, J = 7.36 Hz), 7.30 (2H, t, J = 7.4 Hz), 7.23-7.18 (3H, m), 6.97-6.93 (2H, m), 4.44 (1H, q, J = 7.20 Hz), 2.75-2.65 (2H, m), 2.16-2.10 (2H, m). ESI-MS [M + H]+ = 328.2 [M − H]− = 326.0 Yield 54.2%
1H NMR (DMSO, 400 MHz) δ = 11.00 (1H, s), 8.22 (1H, d, J = 7.48 Hz), 7.69 (1H, d, J = 6.16 Hz), 7.35- 7.21 (6H, m), 7.07 (1H, t, J = 7.60 Hz), 3.84 (1H, t, J = 4.72 Hz), 3.15 (2H, dd, J = 6.00, 13.84 Hz), 2.88 (2H, dd, J = 8.08, 13.80 Hz). ESI-MS [M + H]+ = 321.0 [M − H]− = 318.9 Yield 5.3%
1H NMR (DMSO, 400 MHz) δ = 10.48 (1H, s), 8.59 (1H, s), 7.93 (1H, t, J = 5.68 Hz), 7.10 (1H, d, J = 8.84 Hz), 7.03 (1H, s), 6.83 (1H, s), 6.59 (1H, d, J = 8.60 Hz), 4.36 (1H, t, J = 5.00 Hz), 3.46-3.41 (1H, m), 3.11 (1H, dd, J = 5.32, 8.88 Hz), 2.72 (1H, t, J = 6.84 Hz), ESI-MS [M + H]+ = 290.2 [M − H]− = 288.0 Yield 38.0%
1H NMR (DMSO, 400 MHz) δ = 10.80 (1H, s), 7.95 (1H, t, J = 5.72 Hz), 7.55 (1H, d, J = 7.84 Hz), 7.33 (1H, d, J = 8.08 Hz), 7.15 (1H, s), 7.06 (1H, t, J = 5.80 Hz), 6.97 (1H, t, J = 7.96 Hz), 3.38-3.32 (3H, m), 3.11 (1H, t, J = 5.24 Hz), 2.82 (2H, t, J = 7.36 Hz), 1.70-1.65 (3H, m), 1.42-1.35 (1H, m), 1.23-1.16 (1H, m), 0.88-0.83 (6H, m). ESI-MS [M + H]+ = 274.3 [M − H]− = 272.0 Yield 35.3%
1H NMR (DMSO, 400 MHz) δ = 11.02 (1H, s), 8.28 (1H, d, J = 8.24 Hz), 7.69 (1H, d, J = 7.72 Hz), 7.31-7.15 (6H, m), 7.02 (1H, t, J = 7.56 Hz), ESI-MS [M + H]+ = 335.1 [M − H]− = 332.8 Yield 17.1%
1H NMR (DMSO, 400 MHz) δ = 10.51 (1H, s), 8.97 (1H, t, J = 5.68 Hz), 8.61 (1H, s), 7.80 (1H, d, J = 8.48 Hz), 7.14-7.07 (4H, m), 6.88 (1H, s), 6.60 (1H, d, J = 6.28 Hz), 3.53 (2H, dd, J = 6.72, 14.48 Hz), 2.87 (2H, t, J = 7.80 Hz). Yield 31.7% ESI-MS [M + H]+ = 375.1 [M − H]− = 373.0 Yield 31.7%
1H NMR (DMSO, 400 MHz) δ = 10.83 (1H, s), 8.98 (1H, t, J = 5.52 Hz), 7.79 (1H, d, J = 8.48 Hz), 7.57 (1H, d, J = 7.84 Hz), 7.34 (1H, d, J = 7.28 Hz), 7.19 (1H, s), 7.14- 7.05 (3H, m), 6.98 (1H, t, J = 6.96 Hz), 3.58 (2H, dd, J = 6.96, 12.88 Hz), 2.97 (2H, t, J = 7.48 Hz). ESI-MS [M + H]+ = 359.0 [M − H]− = 357.0 Yield 82.4%
1H NMR (DMSO, 400 MHz) δ = 10.89 (1H, s), 9.04 (1H, bs), 7.84 (1H, d, J = 5.96 Hz), 7.50 (1H, d, J = 7.96 Hz), 7.33 (1H, d, J = 8.04 Hz), 7.17 (1H, s), 7.13-7.05 (3H, m), 6.96 (1H, t, J = 8.00 Hz), 4.77 (1H, dd, J = 7.60, 13.04 Hz), 3.32- 3.17 (2H, m). ESI-MS [M + H]+ = 417.1 [M − H]− = 415.0 Yield 58.5%
1H NMR (DMSO, 400 MHz) δ = 9.02 (1H, d, J = 7.32 Hz), 7.80 (1H, d, J = 8.96 Hz), 7.31-7.19 (5H, m), 7.14-7.11 (2H, m), 4.78- 4.73 (1H, m), 3.66 (3H, s), 3.29- 3.08 (2H, m). ESI-MS [M − H]− = 378.0 Yield 66.3%
1H NMR (DMSO, 400 MHz) δ = 8.95 (1H, d, J = 7.64 Hz), 7.83 (1H, d, J = 8.36 Hz), 7.30-7.11 (7H, m), 4.72-4.67 (1H, m), 3.21 (1H, dd, J = 4.84, 13.88 Hz), 3.10 (1H, dd, J = 9.00, 13.84 Hz). ESI-MS [M − H]− = 361.8 Yield 88.0%
1H NMR (DMSO, 400 MHz) δ = 9.07 (1H, d, J = 7.36 Hz), 8.05 (1H, s), 7.55 (1H, d, J = 6.24 Hz), 7.31- 7.21 (5H, m), 4.73 (1H, dd, J = 8.32, 13.40 Hz), 4.10 (2H, q, J = 7.08 Hz), 3.21-3.10 (2H, m), 1.16 (3H, t, J = 4.72 Hz). ESI-MS [M + H]+ = 392.0 [M − H]− = 390.0 Yield 58.6%
1H NMR (DMSO, 400 MHz) δ = 10.85 (1H, s), 8.95 (1H, d, J = 7.40 Hz), 7.86 (1H, d, J = 8.36 Hz), 7.54 (1H, d, J = 7.92 Hz), 7.32 (1H, d, J = 7.68 Hz), 7.16-7.11 (3H, m), 7.05 (1H, t, J = 6.96 Hz), 6.96 (1H, t, J = 7.00 Hz), 4.75-4.70 (1H, m), 3.33 (1H, dd, J = 8.04, 14.40 Hz), 3.24 (1H, dd, J = 8.32, 14.84 Hz). ESI-MS [M + H]+ = 401.0 [M − H]− = 403.0 Yield 88.0%
1H NMR (DMSO, 400 MHz) δ = 10.78 (1H, s), 9.40 (3H, bs), 7.58 (1H, d, J = 7.80 Hz), 7.33 (1H, d, J = 8.04 Hz), 7.16 (1H, s), 7.06 (1H, t, J = 7.00 Hz), 6.88 (1H, t, J = 8.12 Hz), 6.36 (2H, d, J = 8.12 Hz), 3.02 (2H, dd, J = 7.24, 15.48 Hz), 2.83 (2H, dd, J = 6.04, 8.48 Hz). ESI-MS [M + H]+ = 297.1 [M − H]− = 294.9 Yield 15.4%
1H NMR (DMSO, 400 MHz) δ = 10.94 (1H, s), 9.22 (1H, d, J = 6.80 Hz), 7.44 (1H, d, J = 7.96 Hz), 7.35 (1H, d, J = 8.12 Hz), 7.15 (1H, s), 7.07 (1H, t, J = 5.96 Hz), 6.97 (1H, t, J = 7.08 Hz), 4.85 (1H, dd, J = 6.04, 12.68 Hz), 3.65 (3H, s), 3.30 (2H, d, J = 5.96 Hz). ESI-MS [M + H]+ = 369.1 [M − H]− = 367.1 Yield 33.0%
1H NMR (DMSO, 400 MHz) δ = 10.91 (1H, s), 9.22 (1H, d, J = 6.92 Hz), 7.50 (1H, d, J = 7.92 Hz), 7.34 (1H, d, J = 8.08 Hz), 7.12 (1H, s), 7.06 (1H, dd, J = 7.04 Hz), 6.95 (1H, dd, J = 6.96 Hz), 6.17 (2H, s), 4.77 (1H, dd, J = 6.12, 12.00 Hz), 3.31 (1H, dd, J = 5.12, 9.24 Hz), 3.25 (1H, d, J = 6.20 Hz), 2.15 (3H, s). ESI-MS [M + H]+ = 355.1 [M − H]− = 353.1 Yield 65.2%
1H NMR (DMSO, 400 MHz) δ = 9.14 (1H, d, J = 6.08 Hz), 7.32- 7.18 (6H, m), 6.18 (2H, s), 4.80 (1H, dd, J = 6.72, 12.76 Hz), 4.14 (2H, q, J = 6.08 Hz), 3.17-3.11 (2H, m), 2.14 (3H, s), 1.16 (3H, t, J = 7.12 Hz). ESI-MS [M + H]+ = 344.0 [M − H]− = 342.1 Yield 61.7%
1H NMR (DMSO, 400 MHz) δ = 9.14 (1H, d, J = 7.08 Hz), 7.31- 7.19 (6H, m), 6.17 (2H, s), 4.75 (1H, t, J = 5.36), 3.20 (1H, dd, J = 5.24, 13.88 Hz), 3.11 (1H, dd, J = 6.72, 13.80 Hz), 2.15 (3H, s). ESI-MS [M + H]+ = 316.2 [M − H]− = 313.8 Yield 80.7%
1H NMR (DMSO, 400 MHz) δ = 9.25 (1H, d, J = 6.68 Hz), 7.31- 7.18 (5H, m), 6.23 (2H, s), 4.53 (1H, dd, J = 5.48, 12.76 Hz), 3.66 (3H, s), 2. 65 (2H, t, J = 1.92 Hz), 2.20-2.07 (5H, m). ESI-MS [M + H]+ = 344.1 [M − H]− = 342.0 Yield 100%
1H NMR (DMSO, 400 MHz) δ = 9.30 (1H, bs), 7.30-7.16 (5H, m), 6.22 (2H, s), 4.48 (1H, bs), 2.64 (2H, t, J = 7.72 Hz), 2.22-2.11 (5H, m). ESI-MS [M + H]+ = 330.2 [M − H]− = 327.9 Yield 36.5%
1H NMR (DMSO, 400 MHz) δ = 9.13 (1H, d, J = 6.96 Hz), 7.33- 7.18 (5H, m), 6.18 (2H, s), 4.83 (1H, dd, J = 5.60 Hz, 11.64 Hz), 3. 68 (3H, s), 2.16 (3H, s). ESI-MS [M + H]+ = 330.1 [M − H]− = 328.1 Yield 58.9%
1H NMR (DMSO, 400 MHz) δ = 11.14 (1H, s), 11.05 (1H, s), 8.29 (3H, s), 8.15 (1H, d, J = 7.96 Hz), 7.73 (1H, d, J = 6.40 Hz), 7.66 (1H, d, J = 7.84 Hz). 7.39-7.35 (2H, m), 7.28 (1H, s), 7.16-7.07 (2H, m), 6.96 (1H, t, J = 7.44), 4.22 (1H, dd, J = 4.80, 7.56 Hz), 3.46 (1H, dd, J = 4.56, 15.16 Hz), 3.27 (1H, dd, J = 7.80, 15.04 Hz). ESI-MS [M + H]+ = 360.0 [M − H]− = 358.0 Yield 26.7%
1H NMR (DMSO, 400 MHz) δ = 10.93 (1H, s), 10.00 (1H, s), 9.00 (1H, d, J = 6.88 Hz), 7.44 (1H, d, J = 7.92), 7.35 (1H, d, J = 8.08 Hz), 7.14 (1H, s), 7.07 (1H, t, J = 6.00 Hz), 6.97 (1H, t, J = 6.04 Hz), 5.79 (2H, s), ESI-MS [M + H]+ = 371.1 [M − H]− = 369.1 Yield 49.0%
1H NMR (DMSO, 400 MHz) δ = 10.90 (1H, s), 9.95 (1H, s), 8.99 (1H, d, J = 6.96 Hz), 7.51 (1H, d, J = 7.92 Hz), 7.33 (1H, d, J = 8.04 Hz), 7.12 (1H, s), 7.06 (1H, t, J = 7.16), 6.94 (1H, t, J = 7.00 Hz), 5.77 (2H, s), 4.74 (1H, dd, J = 6.32, 12.04 Hz), 3.30-3.22 (2H, m). ESI-MS [M + H]+ = 357.2 [M − H]− = 355.0 Yield 65.5%
1H NMR (DMSO, 400 MHz) δ = 10.03 (1H, s), 8.91 (1H, d, J = 7.00 Hz), 7.33-7.18 (5H, m), 5.80 (2H, s), 4.78 (1H, dd, J = 7.00, 12.48Hz), 3.67 (3H, s), 3.19-3.08 (2H, m). ESI-MS [M + H]+ = 332.1 [M − H]− = 330.1 Yield 33.7%
1H NMR (DMSO, 400 MHz) δ = 9.97 (1H, s), 8.91 (1H, d, J = 7.04 Hz), 7.32-7.19 (5H, m), 4.73 (1H, dd, J = 6.72, 12.12 Hz), 3.18 (1H, dd, J = 5.24, 13.92 Hz), 3.10 (1H, dd, J = 6.88, 13.88 Hz). ESI-MS [M + H]+ = 318.2 [M − H]− = 316.1 Yield 76.2%
1H NMR (DMSO, 400 MHz) δ = 10.84 (1H, s), 8.92 (1H, s), 7.60 (1H, d, J = 7.92 Hz), 7.34 (1H, d, J = 8.12 Hz), 7.19 (1H, s), 7.08 (1H, t, J = 8.04 Hz), 6.98 (1H, t, J = 7.00 Hz), 6.17 (2H, s), 3.63 (2H, dd, J = 7.16, 12.84 Hz), 2.97 (2H, t, J = 7.08 Hz), 2.15 (3H, s). ESI-MS [M + H]+ = 311.2 [M − H]− = 308.9 Yield 15.5%
1H NMR (DMSO, 400 MHz) δ = 10.84 (1H, s), 9.78 (1H,s), 8.71 (1H, t, J = 5.44 Hz), 7.60 (1H, d, J = 7.84 Hz), 7.34 (1H, d, J = 8.08 Hz), 7.18 (1H, s), 7.08 (1H, t, J = 7.00 Hz), 6.98 (1H, t, J = 7.04 Hz), 5.78 (2H, s), 3.60 (2H, dd, J = 7.00, 12.92 Hz), 2.95 (2H, t, J = 7.20, 14.32 Hz) ESI-MS [M + H]+ = 313.0 [M − H]− = 310.9 Yield 27.9%
ENaC activation current value, namely, the inward current in ENaC-expressing oocyte was measured as follows.
According to the method described in Motonao Nakamura, Takao Shimizu: Experiment of African clawed frog oocyte, Experimental Medicine Vol. 11, No. 3, 224-232 (1993), oocyte was collected from African clawed frog, microinjected into cRNA of oocyte, and the current value was measured by a Two Electrode Voltage Clamp technique. In the present invention, ND96 (96 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM Hepes, 1.8 mM CaCl2, pH 7.6) was used instead of MBS buffer in the above-mentioned document. Injector NANOLITER 2000 manufactured by World Precision Instruments was used for microinjection, OpusXpress 6000A manufactured by Molecular Devices was used for current value measurement by a Two Electrode Voltage Clamp technique, and Clampfit 10.2 software manufactured by Molecular Devices was used for data analysis.
Human ENaC δ, β and γ subunit genes shown by SEQ ID NOs: 1, 2 and 3, respectively, in the Sequence Listing were each cloned into a plasmid vector, and used as a template DNA for cRNA synthesis. cRNA was synthesized using MEGAscript kit manufactured by Ambion and according to the method of the manual of the manufacturer.
A mixture of the same amount of ENaC δ subunit cRNA (0.4 to 0.8 μg/μL), and β and γ subunits cRNA (0.4 μg/μL) was injected into the oocyte of African clawed frog at 27.6 nL per oocyte, and the oocyte was cultured for 16 hours to 72 hours. Thereafter, the oocyte was set on a measuring apparatus OpusXpress 6000A, the electrodes were inserted into the oocyte, the voltage was clamped at a level lower by −30 mV than the resting membrane potential by a Two Electrode Voltage Clamp technique, and the current value was measured.
To measure the ENaC activation potency, oocyte was stimulated with each test compound, and the minimum effective dose (hereinafter to be abbreviated as MED) of the compound at the time point when the electric current started to increase was measured. In this case, the minimum effective dose of the stimulation compound that increased the current value by not less than 10%, as compared to the current value before addition of the compound and without stimulation, was taken as MED. A smaller MED means higher ENaC activation potency.
Each test compound was directly dissolved in ND96, or first dissolved in dimethyl sulfoxide (DMSO) to 100 mM, and then prepared by diluting the solution with ND96 to a concentration used for the evaluation. The concentration of the test compound in ND96 was prepared to various concentrations within the range of 0.1 nM to 300 μm. DMSO at a concentration contained in the prepared liquid does not influence the current value of ENaC.
ND96 as a perfusion fluid was flowed at a rate of 0.5 mL/min through the measuring apparatus OpusXpress 6000A on which ENaC-expressing oocyte was set, the perfusion fluid was temporarily stopped every 3 minutes to add a test compound solution (0.25 mL) dissolved in ND96 at a rate of 0.5 mL/minute. In this way, the test compound was contacted with ENaC-expressing oocyte, and whether ENaC is activated by the compound was examined by measuring the current value. After addition of the test compound, feeding of the perfusion fluid was resumed to wash away the compound to be ready for the next addition of the compound.
The compound was added successively from a low concentration to a high concentration to stimulate ENaC-expressing oocyte. The concentrations actually added was, for example, 1 nM, 3 nM, 10 nM, 30 nM, 100 nM, 300 nM; 1 μM, 3 μM, 10 μm, 30 μm, 100 μm, 300 μm; 1.2 μM, 6 μM, 30 μm, 150 μm of one test compound. The compound was added from low concentrations to high concentrations at 3 minute intervals. The first compound was added from a low concentration to a high concentration, then the second and the third compounds were similarly added successively to the same oocyte, and changes in the current value were measured. The current value was continuously measured from the start to the end of the experiment.
The concentration of the compound that increased the current value by not less than 10% for the first time after compound addition, as compared to the current value immediately before addition of the compound and without stimulation, was taken as MED. As one embodiment of the MED measurement, a graph of the current value when ENaC-expressing oocyte was stimulated with the compound of Example 32, is shown in
MED values of other representative Example compounds, which were obtained by measuring in the same manner, are shown in the following Table 2.
Thus, the compounds of the present invention were confirmed to have an ENaC activating action.
The strength of the salty taste enhancing activity of the compound of Example 35 was examined by a quantitative sensory evaluation test.
The quantitative sensory evaluation test was performed as follows. The strength of the salty taste enhancing activity when the compound of Example 35 as a sample was mixed at 0.00005 to 0.0040 g/dl with distilled water containing sodium chloride (0.5 g/dl) was measured. Distilled water containing sodium chloride at 0.55 g/dl, 0.60 g/dl, 0.65 g/dl was used as a comparison target. Using a linear measurement method, the corresponding marks were put down as positions in the straight lines showing the sodium chloride concentrations of 1.0 time (0.50 g/dl), 1.1 times (0.55 g/dl), 1.2 times (0.60 g/dl), 1.3 times (0.65 g/dl). For sensory marks, the positions put down by the panelists were measured, averaged and shown as the salty taste enhancement ratio (times) wherein n=5. The panelists were those who had an experience of developing flavoring of foodstuffs for a total of one year or more, and capable of distinguishing sodium chloride solutions having different concentrations of 0.50 g/dl, 0.55 g/dl, 0.60 g/dl (periodically confirmed). The “initial taste” is the evaluation of the strength of the salty taste for 2 seconds after placing in the mouth, and the “middle and after taste” is a combination of middle taste and after taste, which is the evaluation of the strength of the salty taste after 2 seconds from placing in the mouth. The results are shown in
In addition, the results of ENaC activation current of Example 35 measured in the same manner as in Experimental Example 1 are shown in
Such strong correlation between the salty taste enhancing effect and the ENaC activation current value suggests that at least one part of salty taste acceptance on the tongue is mediated by an ENaC salty taste receptor, and a compound that activates ENaC has a salty taste enhancing activity.
Therefore, the compound of the present invention having an ENaC activating action shown in Table 2 is strongly suggested to have a salty taste enhancing activity.
The strength of the salty taste enhancing activity of the compound of Example 35 when used in combination with known salty taste alternative was examined by a quantitative sensory evaluation test.
The quantitative sensory evaluation test was performed as follows. The strength of the salty taste enhancing activity when the compound of Example 35 (0.001 g/dl) as a sample and potassium chloride (0.325 g/dl) were mixed with distilled water containing sodium chloride (0.5 g/dl) was measured. Distilled water containing sodium chloride at 0.55 g/dl, 0.60 g/dl, 0.65 g/dl, 0.70 g/dl was used as a comparison target. Furthermore, for comparison evaluation, distilled water containing sodium chloride (0.5 g/dl) and mixed with the compound of Example 35 (0.001 g/dl), and distilled water containing sodium chloride (0.5 g/dl) and mixed with potassium chloride (0.325 g/dl) were used as comparison targets. Using a linear measurement method, the corresponding marks were put down as positions in the straight lines showing the sodium chloride concentrations of 1.0 time (0.50 g/dl), 1.1 times (0.55 g/dl), 1.2 times (0.60 g/dl), 1.3 times (0.65 g/dl), 1.4 times (0.70 g/dl). For sensory marks, the positions put down by the panelists were measured, averaged and shown as the salty taste enhancement ratio (times) wherein n=5. The panelists were those who had an experience of developing flavoring of foodstuffs for a total of one year or more, and capable of distinguishing sodium chloride solutions having different concentrations of 0.50 g/dl, 0.55 g/dl, 0.60 g/dl, 0.65 g/dl, 0.70 g/dl (periodically confirmed). The “initial taste” is the evaluation of the strength of the salty taste for 2 seconds after placing in the mouth, and the “middle and after taste” is a combination of middle taste and after taste, which is the evaluation of the strength of the salty taste after 2 seconds from placing in the mouth. The results are shown in
The results have confirmed that the compound of the present invention has a superior effect even when used in combination with existing salty taste alternatives.
The compounds of the present invention provide a compound a strong salty taste enhancing effect, which is useful for a salty taste enhancer for food and drink, and the like.
Where a numerical limit or range is stated herein, the endpoints are included. Also, all values and subranges within a numerical limit or range are specifically included as if explicitly written out.
As used herein the words “a” and “an” and the like carry the meaning of “one or more.”
Obviously, numerous modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that, within the scope of the appended claims, the invention may be practiced otherwise than as specifically described herein.
All patents and other references mentioned above are incorporated in full herein by this reference, the same as if set forth at length.
This application is a continuation of International Patent Application No. PCT/JP2010/067005, filed on Sep. 29, 2010, which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/JP2010/067005 | Sep 2010 | US |
| Child | 13852434 | US |
