MXPA00011184A - Thienopyridine compounds, their production and use - Google Patents

Thienopyridine compounds, their production and use

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Publication number
MXPA00011184A
MXPA00011184A MXPA/A/2000/011184A MXPA00011184A MXPA00011184A MX PA00011184 A MXPA00011184 A MX PA00011184A MX PA00011184 A MXPA00011184 A MX PA00011184A MX PA00011184 A MXPA00011184 A MX PA00011184A
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Mexico
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substituted
group
compound
salt
alkoxy
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MXPA/A/2000/011184A
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Spanish (es)
Inventor
Shuichi Furuya
Nobuo Choh
Nobuhiro Suzuki
Takashi Imada
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Nobuo Choh
Shuichi Furuya
Takashi Imada
Nobuhiro Suzuki
Takeda Chemical Industries Ltd
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Application filed by Nobuo Choh, Shuichi Furuya, Takashi Imada, Nobuhiro Suzuki, Takeda Chemical Industries Ltd filed Critical Nobuo Choh
Publication of MXPA00011184A publication Critical patent/MXPA00011184A/en

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Abstract

The compound of the present invention possesses excellent gonadotropin-releasing hormone antagonizing activity, and is useful for preventing or treating sex hormone-dependent diseases, e.g. , sex hormone-dependent cancers (e.g., prostatic cancer, uterine cancer, breast cancer, pituitary tumor), prostatic hypertrophy, hysteromyoma, endometriosis, precocious puberty, amenorrhea syndrome, multilocular ovary syndrome, pimples etc., or as a pregnancy regulator (e.g., contraceptive), infertility remedy or menstruation regulator.

Description

DESCRIPTION Thienopyridine Compounds, Their Production and Use TECHNICAL FIELD The present invention relates to thieno [2, 3-b] pyridine derivatives, which exhibit antagonistic activity to gonadotropin releasing hormone (GnRH), its production and use.
BACKGROUND TECHNIQUE The secretion of the hormone of the hypophysial anterior lobe is regulated by the peripheral hormone secreted by each target organ and the promotion of the secretion or suppression of the secretion of the hormone secreted by the hypothalamus, which is the superior center to the anterior lobe. pituitary, and this group of hormones will be referred to hereinafter to this group of hormones as hypothalamic hormone in this specification. To date, nine hypothalamic hormones have been identified, for example, the hormone releasing the thyroid stimulating hormone (THR), and the gonadotropin releasing hormone [GnHR, also known as the hormone releasing the luteinizing hormone (LH-RH)], etc. It is surmised that these hypothalamic hormones exhibit their Ref. No. 124427 hormonal actions, etc. via receptors that are added are present in the anterior hypophysial lobe, and are underway, analysis of specific receptor genes for these hormones, including humans. Antagonists or agonists that act specifically and selectively on these receptors, should therefore regulate the action of the hypothalamic hormones and, therefore, regulate the secretion of the hormone of the anterior hypophysial lobe. As a result, it is expected that such antagonists or agonists, prevent or treat diseases that depend on this hormone of the anterior hypophysial lobe.
Known compounds possessing GnRH-antagonistic activity include linear peptides derived from GnHR (USP 5,140,009 and USP 5, 171,835), a cyclic hexapeptide derivative (JP-A-61-191698), a bicyclic peptide derivative [ Journal of Medicinal Chemistry, Vol. 36, pp. 3265-3273 (1993)], and so on. Non-peptide compounds, which possess GnRH antagonizing activity, include the compounds described in WO 95/28405, WO 97/14697, WO 97/14682, WO 97/41126 and so on.
Peptide compounds have a large number of problems to be solved, with respect to ABSORBABILITY, dosage form, dosage volume, drug stability, sustained action, metabolic stability, etc. There is a strong demand for an oral GnRH antagonist, especially one based on a non-peptide compound, which has an excellent therapeutic effect on hormone-dependent cancers, for example prostate cancer, endometriosis, precocious puberty, etc. And that they do not show transitory hypophysial gonadotropic action (acute action).
BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagrammatic representation of the relieving effect of the suppression of LH plasma concentrations in castrated monkeys, after oral administration of the compound of the following Example 1-2, in which ••••• •• represents the control (1) (vehicle), ••• ••• represents the control (2) (vehicle), ... [3 ... represents the control (3) (vehicle), -p- represents the compound (1) [the test administered to the animal the compound of Ex. No. 1-2] and -p- represents the compound (2) [the test compound administered to the animal, the compound of ex. No. 1-2].
DESCRIPTION OF THE INVENTION The present inventors produced various thienopyridine derivatives, investigated their action, and found that some compounds possess excellent GnRH antagonizing activity. The inventors have conducted other investigations based on these discoveries, and developed the present invention.
Accordingly, the present invention relates to: [1] a compound of the formula: wherein R1 represents an alkyl group of C? -, which may be substituted, a cycloalkyl group of C3-, which may be substituted, an alkoxyamino group of C? _6, which may be substituted or a hydroxyamino group, which may be substituted; and R2 represents an alkyl group of O.- ?, which may be substituted or a phenyl group, which may be substituted; When R1 is an unsubstituted C1-7 alkyl group, then R2 is a C1-7 alkyl group or a substituted phenyl group, or a salt thereof [sometimes called hereafter for brevity as the compound (I) ]: [2] a compound of (I) above or a salt thereof, wherein R1 is (1) a C1-7 alkyl group, which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) C 1-7 acyloxy, (iii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C 1-6 alkoxycarbonyl, benzyloxycarbonyl, C 1-3 acyl , C 1 -C 3 alkylsulfonyl and C 1 -C 3 alkyl, (iv) C 1-10 alkoxy, which may be substituted by 1 to 3 secstituted substituents of the group consisting of C 3 7 cycloalkyloxycarbonyl and C 1 -C 7 alkoxy 3 and (v) C? -6-carbonyl alkoxy, (2) a C3-7 cycloalkyl group, which may be substituted by 1 to 3 substituents selected from the group consisting of (i) hydroxy, (ii) C? _7 acyloxy, (iii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C?-6-carbonyl alkoxy, benzyloxycarbonyl, C ac acyl? _3, C ?_3 alkylsulfonyl and C ?_alkyl, (iv) C 1-10 alkoxy, which may be substituted by 1 to 3 substituents selected from the group consisting of C 3-7 cycloalkyloxy-carbonyl and C-alkoxy; -3 and (v) C6-6-carbonyl alkoxy, (3) an alkoxyamino group of C6-6, which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) ) C 1-7 acyloxy, (iii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C?-6-carbonyl alkoxy, benzyloxycarbonyl, C 1-3 acyl, C 1-3 alkylsulfonyl and C 1 -3 alkyl, (iv) C 1 0 alkoxy, which may be substituted by 1 to 3 substituents selected from the group consisting of C 3 7 -carbonyl cycloalkyloxy and C 1-3 alkoxy and (v) C6-6-carbonyl alkoxy, or (4) a hydroxyamino group, which may be substituted by 1 or 2 substituents selected from the group consisting of (i) C1-7 acyloxy, (ii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C6-C6-alkynyl, benzyloxycarbonyl, C1-3 acyl, C1-3 alkylsulfonyl and C3_3 alkyl, (iii) alkoxy C1-10, which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkyloxycarbonyl and C1-3alkoxy and (iv) C6-6alkyl; and R2 is (1) an alkyl group of C7_7 which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) C1-7 acyloxy, (iii) amino, to which it may be substituted by 1 or 2 substituents selected from the group consisting of C?-6-carbonyl alkoxy, benzyloxycarbonyl, C?-3 acyl, C 1-3 alkylsulfonyl and C? _3 alkyl, (iv) C 1 - alkoxy 10, which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkyloxy-carbonyl and C1-3.alkoxy and (v) Ci-β-carbonyl-alkoxy, or (2) a group phenyl, which may be substituted by 1 to 5 substituents selected from the group consisting of halogen, C 1-3 alkyl and C 1 -3 alkoxy; [3] a compound of (1) above or a salt thereof, wherein R1 is a C1-7 alkyl group, which may be substituted or a C3-7 cycloalkyl group, which may be substituted; [4] a compound of (1) above or a salt thereof, wherein R1 is a substituted, branched C3-7 alkyl group or a substituted C3-7 cycloalkyl group; [5] a compound of (1) above or a salt thereof, wherein R1 is an alkyl group of C? -? substituted by hydroxy, or a C3-7 cycloalkyl group substituted by hydroxy; [6] a compound of (1) above or a salt thereof, wherein R1 is a substituted C3-7 cycloalkyl group; [7] a compound of (1) above or a salt thereof, wherein R1 is a cyclopropyl group, which may be substituted by hydroxy; [8] a compound of (1) above or a salt thereof, wherein R2 is a branched C3_7 alkyl group, which may be substituted; [9] a compound of (1) above or a salt thereof, wherein R2 is a phenyl group, which may be substituted; [10] a compound of (1) above or a salt thereof, wherein Rz is a phenyl group; [11] a compound of (1) above or a salt thereof, wherein R1 is a cycloalkyl group of C3-7 and R2 is an alkyl group of C? -6, '[12] a compound of (1) above or a salt thereof, wherein R1 is (1) a C1-4 alkyl group, substituted 1 or 2 hydroxy, (2) a C3_7 cycloalkyl group substituted by hydroxy, or (3) an alkoxyamino group of C ?-3; Y R2 is an isopropyl group or a phenyl group; [13] a compound of (1) above or a salt thereof, wherein R1 is (1) a C1-7 alkyl group, which may be substituted by i2 substituents selected from the group consisting of hydroxy, alkylcarbonyl of C? -3, amino, benzyloxycarbonylamino, C? _3 alkoxy, C? _3 alkoxy-C1-3 alkoxy and C? -3-carbonyl alkoxy, (2) a C3-7 cycloalkyl group, which may be substituted by a hydroxy or a C 1 -3-carbonyloxy alkyl, or (3) an alkoxyamino group of C 3 -3; and Rz is (1) an isopropyl group, which may be substituted by a hydroxy or (2) a phenyl group; [14] 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4-oxothiene [2, 3 -b] pyridine or a salt thereof; [15] 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- (3-hydroxy-2-methylpropionylamino) phenyl ] -4-oxothieno [2, 3-b] pyridine or a salt thereof; [16] 3- (N-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2, 3-b] pyridine or a salt thereof; [17] 3- (N-Benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2, 3-b] pyridine or a salt thereof; [18] a process for the production of a compound of [1] above or a salt thereof, which comprises the reaction of a compound of the formula: wherein R2 represents an alkyl group of C? _7, which may be substituted or a phenyl group, which may be substituted, or a salt thereof [sometimes called for brevity as compound (II)], with i) a compound of the formula: RlaC00H wherein Rla represents an alkyl group of C? _7, which may be substituted or a C3-7 cycloalkyl group, which may be substituted, or a salt thereof or a reagent derived therefrom; or ii) carbonyldiimidazole, phosgene or a chloroformate, followed by reaction with a compound of the formula: RlbH wherein R 1b represents an alkoxyamino group of C 6 -6, which may be substituted or a hydroxyamino group, which may be substituted, or a salt thereof; [19] a process for the production of a compound of [3] above or a salt thereof, which comprises the reaction of a compound (.II) with a compound of the formula: RlaCOOH wherein Rla represents an alkyl group of C? _7, which may be substituted or a cycloalkyl group, which may be substituted, or a salt thereof or a reagent derived therefrom; [20] a pharmaceutical composition, which comprises a compound of (1) above or a salt thereof; [21] a pharmaceutical composition of (20) above, which is to antagonize gonadotropin releasing hormone; [22] a pharmaceutical composition of (21) above, which is to prevent or treat a sex hormone-dependent disease; [23] a method for antagonizing gonadotropin releasing hormone in a mammal in need thereof, which comprises administering to said mammal an effective amount of a compound (1) above or a salt thereof, with a pharmaceutically acceptable excipient, carrier or diluent; [24] the use of a compound (1) above or a salt thereof, for the manufacture of a pharmaceutical composition for antagonizing the gonadotropin releasing hormone, and so on.
Each of the symbols in the formulas above is described hereafter, in more detail.
The "C 1-7 alkyl group" of the "Ci-7 alkyl group, which may be substituted", for R 1 includes, for example, a straight chain C 1-7 alkyl group, such as methyl, ethyl, propyl , butyl, pentyl, hexyl, heptyl, etc .; a branched chain C3_7 alkyl group, such as, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, neopentyl, etc. And so on. Among others, the preferred is a branched C3-7 alkyl group. More preferably it is isopropyl.
The "substituents" of the "C 1-7 alkyl group, which may be substituted", for R 1 include, for example, (i) hydroxy, (ii) C 1-7 acyloxy (e.g., alkylcarbonyloxy) C? -6, such as acetoxy, propionyloxy, etc., benzyloxy, etc.), (iii) amino, which may be substituted by 1 02 substituents selected from the group consisting of C? _6 alkoxycarbonyl (e.g. methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, etc.), benzyloxycarbonyl, C ?3 acyl (for example, C? _-carbonyl alkyl such as acetyl, propionyl, etc.), C? _3 alkylsulfonyl (e.g. methanesulfonyl, etc.) and C? -3 alkyl. { for example, methyl, ethyl, etc.), etc. [e.g., amino, methoxycarbonylamino, ethoxycarbonylamino, tert-butoxycarbonylbenzyloxycarbonylamino, acetylamino, methanesulphonylamino, methylamino, dimethylamino, etc.), (iv) Ci -io alkoxy (preferably C? -4), which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7 cycloalkylcarbonyl (e.g., cyclohexylcarbonyloxy, etc.) and C 1-3 alkoxy (e.g., methoxy, ethoxy, etc.) [e.g. methoxy, ethoxy, propoxy, terbutoxy, cyclohexyloxycarbonyloxy-1-ethoxy, methoxymethoxy, ethoxymethoxy, etc.], (v) C? -6-carbonyl alkoxy (eg, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, etc.), and so on . Among others, the preferred one is hydroxy.
The "C 1-7 alkyl group" may have 1 to 5, preferably 1 to 3 substituents as mentioned above, at the possible positions and, when the number of substituents is two or more, these substituents may be the same or different ones of others.
The "C3-7 cycloalkyl group" of the "C3_7 cycloalkyl group, which may be substituted", for R1 includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. Among others, it is preferably cyclopropyl.
The "substituents" of the "C3_7 cycloalkyl group, which may be substituted", for R1, are the same as those mentioned above for the "substituents" of the "C7_7 cycloalkyl group, which may be substituted", for R1. the number of substituents is 1 to 3. when the number of substituents is two or more, those substituents may be the same or different from each other.
The "C6-6 alkoxyamino group" of the "C6-C6 alkoxyamino group, which may be substituted" for R1 includes, for example, C- or β-diaminoalkylamino, (eg, methoxyamino, ethoxyamino) , dimethoxyamino, diethoxyamino, ethoxymethoxyamine, etc.), etc. Among others, the preferred is C 1 -3 mono-alkoxyamino (eg, methoxyamino, etc.).
The "substituents" of the "C 1 -C 6 alkoxyamino group, which may be substituted", for R 1 and its number, are the same as those mentioned for the "substituents" of the C 1-7 alkyl group, which may be substituted "for R1, when the number of substituents is two or more, these substituents may be the same or different from each other: the" C6-6 alkoxy "or the" amino group nitrogen atom "of the Ci-alkoxyamino group -ß, may be replaced by the "substituents" above.
Such "C1-6 alkoxyamino group, which may be substituted" is exemplified by methoxyamino, N-methyl-N-methoxyamino, n-ethyl-N-methoxyamino, ethoxyamino, dimethoxyamino, diethoxyamino, ethoxymethoxyamino, etc. Preferred is C 1-3 alkoxyamino, N-C 1-3 alkyl N-alkoxyamino, etc.
The "substituents" of the hydroxyamino group, which may be substituted "for R1, may be located in the" hydroxy group "of the hydroxyamino group or the" nitrogen atom of an amino group "of the hydroxyamino group. "hydroxy group" include, for example, (i) C1-7 acyloxy (eg, C6-6-carbonyloxy alkyl, such as acetoxy, propionyloxy, etc., benzyloxy, etc.), (ii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C6-6-carbonyl alkoxy (e.g., methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, etc.), benzyloxycarbonyl, C1-3 acyl (e.g. , Ci-2-carbonyl alkyl, such as acetyl, propionyl, etc.), C 1-3 alkylsulfonyl, (e.g., methanesulfonyl, etc.) and C 1-3 alkyl (e.g., methyl, ethyl, etc.). ) . etc. [for example; amino, methoxycarbonylamino, ethoxycarbonylamino, tert-butoxycarbonylbenzyloxycarbonylamino, acetylamino, methanesulfonylamino, methylamino, dimethylamino, etc.], (ii) C 1-10 alkoxy (preferably C 1-4), which may be substituted by 1 to 3 substituents selected from the group consisting of a group consisting of cycloalkyloxycarbonyl of C3-7 (for example, cyclohexyloxycarbonyloxy, etc.) and C1-3alkoxy (for example, methoxy, ethoxy, etc.) [for example; methoxy, ethoxy, propoxy, tert-butoxy, cyclohexyloxycarbonyloxy, methoxymethoxy, ethoxymethoxy, etc.], and so on. The number of substituents is 1 to 5, preferably 1 to 3. When the number of substituents is two or more, those substituents may be the same or different from each other. Such "substituents" on the "amino group nitrogen atom" include, for example, (1) each of the groups described in subparagraphs (i) to (iii) above and (2) C? _6 alkyl ( for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, etc. The number of substituents is from 1 to 5, preferably from 1 to 3. When the number of substituents is two or more, these substituents may be the same or different from each other.
Preferred examples of the "hydroxyamino group, which may be substituted" include N-alkyl-6-N-hydroxyamino-alkyl (eg, N-methyl-N-hydroxyamino, N-ethyl-N-hydroxyamino, etc.) and so on The most preferred is N-alkyl of C1- .3-N-hydroxyamino, etc.
The "C? _7 alkyl group" of the "Ci-7 alkyl group, which may be substituted" for R2 includes, for example, an alkyl group of C? -? straight or branched chain, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, etc. Among others, the preferred is C1-3 alkyl (eg, methyl, ethyl, propyl, isopropyl, etc.). The most preferred is isopropyl.
The "substituents" of the "C" _7 alkyl group which may be substituted "by R2 and their number are the same as those mentioned above for the" substituents "of the" C1_7 alkyl group, which may be substituted " for R1. When the number of substituents is two or more, these substituents may be the same or different from each other.
The "substituents" of the "phenyl group, which may be substituted" for R 2, include, for example, halogen (e.g., fluorine, chlorine, bromine, iodine, etc.), C 1-3 alkyl (e.g., methyl) , ethyl, propyl, isopropyl, etc.), C1-3 alkoxy (eg, methoxy, ethoxy, propoxy, isopropoxy, etc.), etc. Among others, the preferred one is halogen, the most preferred being fluorine.
The "phenyl group" may have from 1 to 5, preferably from 1 to 3 s, as mentioned above, in the possible positions and, when the number of substituents is two or more, those substituents may be the same or different ones. of others.
R1 is preferably a substituted, branched C3_7 alkyl group or a substituted C3-7 cycloalkyl group, more preferably a C7_7 alkyl group substituted by hydroxy or a C3_7 cycloalkyl group, substituted by hydroxy. Among others, the preferred one is a C3-7 cycloalkyl group, substituted. Also, an alkyl group of C? -3 / which may be substituted by hydroxy, a C3-7 cycloalkyl group, which may be substituted by a hydroxy, C1-3 mono-alkoxyamino, C1- N-alkyl 3-N-hydroxyamino, hydroxyamino, is preferred.
Especially preferred R1 is (i) cyclopropyl, which may be substituted by a hydroxy or (ii) methoxyamino, etc. The most preferred is cyclopropyl, which may be substituted by a hydroxy group.
R 2 is preferably a C 1-7 alkyl group, which may be substituted. Most preferred is a C1-3 alkyl group, which may be substituted by a hydroxy group. Isopropyl is especially preferred. Phenyl is also preferred.
Preferred examples of the compound (I), include a compound wherein, R 1 is an alkyl group of C 3 -3 / which may be substituted by a hydroxy group, a C 3-7 cycloalkyl group, which may be substituted by a hydroxy group or a mono-alkoxyamino group of C? -3; and R2 is an alkyl group of C? -3 or a salt thereof.
The most preferred is a compound, wherein, R1 is (1) an alkyl group of C? -4, substituted by 1 or 2 hydroxy, (2) a C3-7 cycloalkyl group, substituted by hydroxy, or (3) a C1-3 alkoxyamino group; and R2 is an isopropyl group or a phenyl group, or a salt thereof.
A compound wherein R 1 is (1) a C 1-7 alkyl group, which may be substituted by 1 or 2 substituents selected from the group consisting of hydroxy, C 3 -carbonyl, amino, benzyloxycarbonylamino, alkoxy, C3-C3-alkoxy and C3-carbonyl-alkoxy, (2) a C3_7 cycloalkyl group, which may be substituted by a hydroxy group or a C3-carbonyloxy alkyl, or (3) a C1-3 alkoxyamino group; and R2 is (1) an isopropyl group, which may be substituted by a hydroxy group or (2) a phenyl group, or a salt thereof, is also preferred.
As compound (I), specifically mentioned are 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4 -oxothieno [2, 3-b] pyridine or a salt thereof, 5-benzoyl-3- (N-benzyl-N-methylaminomethyl) -7- (2,6-difluorobenzyl) -4,7-dihydro-4- oxo-2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] thieno [2, 3-b] pyridine, 5- (4-fluorobenzoyl) -3- (N-benzyl-N-methylaminomethyl) -7- (2,6-difluorobenzyl) -4,7-dihydro-4-oxo-2- (4-cyclopropanecarbonylaminophenyl) thieno [2,3-b] pyridine, 3- (N-benzyl-N-methylaminomethyl) -4,7 -dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -4-oxothieno [2, 3-b] pyridine, 3- (N -benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-N 1 -methoxyureidophenyl) -4-oxothieno [2,3-b] pyridine, 3- (N-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] - 4-oxothieno [2, 3-b] pyridine, (R) -4,7-dihydro-2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3 - (N-benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2,3-b] pyridine, 4,7-dihydro-2- [4- (2-hydroxy-2-methylpropionylamino) phenyl] - 7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2,3-b] pyridine, 4,7-dihydro-2- [4- ( 3-hydroxy-3-methylbutyrylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine, ( R) -4,4-dihydro-2- [4- (2,3-dihydroxypropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyri-1 4-oxothieno [2, 3-b] pyridine, 3- (N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-2- [4- [ (1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2, 3-b] pyridine, and salts thereof.
The compound (I) can be produced in any manner known per se, for example, according to the methods described in WO 95/28405 or methods analogous thereto. Mentioned concretely are, the following Production Method 1 and Production Method 2.
Production method 1: The compound (I) is produced by the reaction of the compound (II) with a compound of the formula, RlaCOOH, where Rla represents a C1-7 alkyl group, which may be substituted or a cycloalkyl group of c3_7, which may be substituted, or a salt thereof or a reagent derived therefrom [sometimes called hereafter, for brevity as compound (III)].
The "C1-7 alkyl group, which may be substituted" and the "C3-7 cycloalkyl group, which may be substituted" for Rla, are the same as those mentioned above for the "C1-7 alkyl group". , which may be substituted "and the" cycloalkyl group of C3-7, which may be substituted "by R1, respectively.
The "derivatized reagent" of the "compound of the formula: RlaCOOH, or a salt thereof or a reagent derived therefrom" includes, for example, a compound of the formula: RlaCOY wherein, Y represents halogen and R is the same as was defined above.
The amount of compound (III) to be subjected to reaction is about 3 moles, relative to one mole of compound (II).
This reaction is also carried out in the presence of a base. The "base" is exemplified by inorganic bases, such as sodium carbonate, sodium hydrogen carbonate, potassium carbonate, potassium carbonate hydroxide, sodium hydroxide, potassium hydroxide and thallium hydroxide, and organic bases such as triethylamine and pyridine, etc.
The amount of the "base" is about 1 to 10 moles, preferably about 2 to 5 moles, relative to one mole of the compound (II).
In the case where a compound of the formula is used; RlaCOOH or a salt thereof, a condensing reagent is employed, which is generally used in peptide chemistry, such as benzotronazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBop), to form an amide bond.
This reaction is advantageously carried out in a solvent inert to the reaction. Examples of the solvent include ethers (e.g., ethyl ether, dioxane, dimethoxyethane, tetrahydrofuran, etc.), aromatic hydrocarbons (e.g., benzene, toluene, etc.), amides (e.g., dimethylformamide, dimethylacetamide, etc.) . halogenated hydrocarbons (for example, chloroform, dichloromethane, etc.). etc.
The reaction temperature is usually about 0 to 150 ° C, preferably at room temperature (about 15 to 25 ° C). The reaction time is usually around 1 to 12 hours.
The compound (II) can be produced in any manner known per se, for example, by the methods described in WO 95/28405, or methods analogous thereto.
The compound (I) wherein R 1 is an alkoxyamino group of C 6 -6, which may be substituted or a hydroxyamino group, which may be substituted, may be produced according to the method of the following, Production method 2.
Production method 2: The compound (I) is produced by the reaction of the compound (II) with carbonyldiimidazole (N, N'-carbonyldiimidazole; CDI), phosgene (monomer, dimer or trimer) or a chloroformate, followed by the reaction with a compound of the formula: RlbH, wherein Rlb represents an alkoxyamino group of C? -6, which may be substituted or a hydroxyamino group, which may be substituted, or a salt thereof [sometimes called hereafter for brevity as the compound (IV)].
The "chloroformate" includes, for example, a compound of the formula: C1-C00Y ', wherein Y' represents an alkyl group of C? -β, such as ethyl chloroformate, etc.
The "alkoxyamino group of C? _6, which may be substituted" and the "hydroxyamino group, which may be substituted", by Rlb, are the same as those mentioned above for the "C? -6 alkoxyamino group, the which may be substituted "and the" hydroxyamino group, which may be substituted "by R1, respectively.
In the reaction of the compound (II) with carbonyldiimidazole, phosgene or chloroformates, the carbonyldiimidazole, phosgene or the chloroformates are used in an amount of about 1 to 3 moles, relative to one mole of the compound (II).
This reaction is advantageously carried out in a solvent inert to the reaction. Examples of the solvent include ethers (for example ethyl ether, dioxane, dimethoxyethane, tetrahydrofuran, etc.), aromatic hydrocarbons (for example, benzene, toluene, etc.), amides (for example, dimethylformamide, dimethylacetamide, etc.) , halogenated hydrocarbons (for example, chloroform, dichloromethane, etc.), etcetera.
The reaction temperature is usually about 0 to 150 ° C, preferably at room temperature (about 15 to 25 ° C). The reaction time is usually around 1 to 36 hours.
This reaction is also carried out in the presence of a base. The "base" is exemplified by organic bases, such as sodium carbonate, sodium hydrogen carbonate, potassium carbonate, potassium hydrogen carbonate, sodium hydroxide potassium hydroxide and thallium hydroxide and organic bases, such as triethylamine and pyridine, etc.
The amount of the "base" is about 2 to 20 moles, preferably about 5 to 12 moles, relative to one mole of the compound (II).
The following reaction with the compound (IV) can be carried out under the same conditions of the above reaction of the compound (II) with carbonyldiimidazole, phosgene or a chloroformate. The amount of the compound (IV) is from about 2 to 20 moles, preferably from about 5 to 10 moles, relative to one mole of the compound (II).
The reaction temperature is usually around 0 to 150 ° C, preferably at room temperature (about 15 to 25 ° C). The reaction time is usually around 1 to 6 hours.
The compound (I) of the present invention can be isolated and purified by ordinary means of separation, such as recrystallization, distillation and chromatography, etc. When the compound of the formula (I) is obtained in the free form, it can be converted to a salt by methods known per se or methods analogous thereto. When the compound (I) is obtained in the salt form, it can be converted to the free form or to another salt, by methods known per se or methods analogous thereto.
The salts of the compound (I) are preferably the physiologically acceptable acid addition salts. Such salts include, for example, salts with inorganic acids (for example, hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid) and physiologically acceptable acid addition salts with organic acids (for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid). When the compound (I) has an acidic group, it can form a pharmaceutically acceptable salt with an inorganic base (for example, alkali metals, such as sodium, potassium, calcium and magnesium, alkaline earth metals, ammonia) or an organic base (e.g. , trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethylamine, dicyclohexylamine, NN '-dibenzylethylenediamine).
The compound (I) can be a hydrate, or a non-hydrate. The hydrate is exemplified by monohydrates, sesquihydrate and dihydrate.
When the compound (I) is obtained as a mixture (racemate) of optically active configurations, can be solved to the forms (R) - and (s) - by conventional optical resolution techniques.
The compound (I) of the present invention (hereinafter also referred to as "the compound of the present invention") possesses excellent GnRH antagonizing activity and low toxicity. In short, it is excellent in its oral absorption capacity, its ability to maintain action, stability and pharmacokinetics.
In addition, it can be easily produced. The compound of the present invention can therefore be used safely in a mammal (e.g., human, monkey, bovine, horse, dog, cat, rabbit, rat, mouse) for the prevention and / or treatment of diseases dependent on female hormones, diseases due to excess of these hormones, etc., by suppressing the secretion of gonadotropin, through its antagonistic action of the GnRH receptor to control the concentrations of sex hormones in the blood.
For example, the compound of the present invention is useful for the prevention and / or treatment of sex hormone-dependent cancers (e.g., prostate cancer, uterine cancer, breast cancer, pituitary tumor, etc.), prostatic hypertrophy , hysteromyoma, endometriosis, precocious puberty, amenorrhea, premenstrual syndrome, multilocular ovary syndrome, pimples, etc. The compound of the present invention is also useful for the regulation of reproduction in men and women (e.g., pregnancy regulators, menstrual cycle regulators, etc.). The compound of the present invention is also used as a male or female contraceptive, or as an inducer of female ovulation. Based on its rebound effect after being removed, the compound of the present invention can be used to treat infertility.
In sum, the compound of the present invention is useful for the regulation of animal estrus, the improvement of the quality of meat, and the promotion of animal growth in the field of animal husbandry. The compound of the present invention is also useful as a promoter of fish roe deposition.
Although the compound of the present invention can be used alone, it is effective to use it in combination with an antiandrogen agent or an anti-estrogen, non-steroidal or steroidal agent. The compound of the present invention can also be used to suppress the transient rise in blood of testosterone concentrations (spreading phenomenon), observed in the administration of a superagonist, such as Leuprorelin Acetate. The compound of the present invention can be used in combination with a superagonist, such as leuprorelin acetate, gonadrelin, buserelin, triptorelin, goserelin, nafarelin, histrelin, deslorelin, meterelin, lecirelin, and the like. Among others, the preferred one is leuprorelin acetate. The compound of the present invention can also be used with a chemotherapeutic agent for cancer. A preferred example of such a combination is the compound of the present invention, in combination with chemotherapeutic agents, such as iforpha ida, UTF, adriamycin, peplomycin and cisplatin for prostate cancer. For breast cancer, the compound of the present invention can be used with chemotherapeutic agents, such as cyclophosphamide, 5-FU, UFT, methotrexate, adriamycin, mitomycin C, and mitoxantrone.
When the compound of the present invention is used for the prevention and / or treatment (as a prophylactic and / or a therapeutic agent for) the above mentioned diseases or is used in the field of animal or fish breeding, it can be administered orally or non-orally, when formulated with a pharmaceutically acceptable carrier, usually in the form of solid preparations, such as tablets, granules and powders for oral administration, or in the form of intravenous, subcutaneous, intramuscular or other injections Suppositories or sublingual tablets for non-oral administration. It can also be sublingually, subcutaneously, intramuscularly or otherwise administered in the form of sustained release preparations of sublingual tablets, microcapsules, etc. Depending on the severity of the symptoms, the age of the subject, sex, weight and sensitivity; duration and administration intervals; properties, administration and type of pharmaceutical preparation; type of active ingredient, etc., the daily dose is not subject to limitation. For use in the treatment of sex hormone-dependent cancers, mentioned above (eg, prosthetic cancer, uterine cancer, breast cancer, pituitary tumor), prostatic hypertrophy, hysteromycin, endometriosis, precocious puberty, etc. The daily dose is usually from about 0.01 to 30 mg, preferably from about 0.02 to 10 mg, and more preferably from 0.1 to 10 mg, per kg of weight of 1 mammal, usually in 1 to 4 divided doses.
The above doses are applicable to the use of the compound of the present invention in the field of raising animals or fish. The daily dose is around 0.01 to 30 mg. Preferably from about 0.1 to 10 mg, per kg of body weight treated, usually in 1 to 3 divided doses.
In the pharmaceutical composition of the present invention, the amount of compound (I) is from about 0.01 to 100% by weight or little more or less, of the total weight of the composition. Pharmaceutically acceptable carriers are various organic and inorganic substances commonly used as pharmaceutical materials, including excipients, lubricants, binders and disintegrators for solid preparations, and solvents, dissolving aids, suspending agents, isotonizing agents, buffers and soothing agents for preparations. liquid. Other pharmaceutical additives, such as preservatives, antioxidants, coloring agents and sweetening agents, may be used, when necessary.
Preferred excipients include, for example, lactose, sucrose, D-mannitol, starch, crystalline cellulose and light salicylic anhydride. Preferred lubricants include, for example, magnesium stearate, calcium stearate, talc and colloidal silica. Preferred binders include, for example, crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose and polyvinylpyrrolidone. Preferred disintegrants include, for example, starch, carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium and carboxymethyl sodium starch. Preferred solvents include, for example, water for injection, alcohol, propylene glycol, magrogol, sesame oil and corn oil. Preferred dissolution aids include, for example, polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethylamine, sodium carbonate and sodium citrate. Preferred suspending agents include, for example, surfers, such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride and glycerol monostearic; and hydrophilic polymers, such as polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose. Isotonizing agents include, for example, sodium chloride glycerol and D-mannitol. Preferred buffers include, for example, buffer solutions of phosphates, acetates, carbonates, citrates, etc. Preferred soothing agents include, for example, benzyl alcohol. Preferred preservatives include, for example, paraoxybenzoic acid, esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and ascorbic acid. Preferred antioxidants include, for example, sulfites and ascorbic acid.
By the addition of suspending agents, dissolution aids, stabilizers, ISOTONIZING agents, preservatives, etc., the compound of the present invention can be prepared as an intravenous, subcutaneous or intramuscular injection, by a commonly known method. In such cases, the compound of the present invention can be dehydrated by freezing when necessary, by a commonly known method. In administration to humans, for example, the compound of the present invention can be administered safely, orally or non-orally, such as a pharmaceutical composition prepared by mixing with a pharmacologically acceptable carrier, excipient or diluent, selected as appropriate.
Such pharmaceutical compositions include oral preparations (e.g., powders, granules, capsules, tablets), injections, drop infusions, external preparations (e.g., nasal preparations, transdermal preparations) and suppositories (e.g., rectal suppositories, vaginal suppositories). .
These preparations can be produced by methods commonly known in the common use of pharmaceutical manufacturing processes.
An injection can be produced by, for example, the preparation of the compound of the present invention, as an aqueous injection together with a dispersing agent (for example, Tween 80, produced by Atlas Powder Company, USA HCO 60, produced by Nikko Chemicals Co., Ltd prolyethylene glycol, carboxymethyl cellulose, sodium alginate), a preservative (e.g., methyl paraben, propyl paraben, benzyl alcohol), an isotonizing agent (e.g., sodium chloride, mannitol, sorbitol, glucose) and other additives, or as an oily injection in solution, suspension or emulsion in a vegetable oil such as olive oil, sesame oil, cottonseed oil or corn oil, propylene glycol or the like.
An oral preparation can be produced by a method commonly known after the addition of an excipient (e.g., lactose, sucrose, starch), a disintegrator (e.g., starch, calcium carbonate), a binder (e.g., starch, gum arabic, carboxymethyl cellulose), polyvinylpyrrolidine, hydroxypropyl cellulose), a lubricant (eg, talc, magnesium stearate, polyethylene glycol 6000) and other additives, and where it is necessary to coat the appearance of the product for the purpose of masking the taste, enteric or sustained release solution by a commonly known method. The coating agents for this purpose include, for example, hydroxypropylmethyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxypropylcellulose, polyoxyethylene glycol, Tween 80, Prulonic F68, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, hydroxymethyl cellulose acetate succinate, Eudragit (produced by Rohm Company, Germany; methylacrylic acid / acrylic acid copolymer) and dyes (for example, iron oxide, titanium dioxide). For an enteric preparation, an intermediate phase between the enteric phase and the phase containing the drug can be provided, for the purpose of separating the two phases by a commonly known method.
An external preparation can be produced by the composition of the compound of the present invention as a solid, semi-solid or liquid composition, by a commonly known method Such a solid composition is produced by, for example, spraying the compound of the present invention. invention such as or in a mixture with an excipient (for example, glycol, mannitol, starch, microcrystalline cellulose), a thickening agent (for example, natural gum, cellulose derivative, acrylic acid polymer) and other additives. produced by the preparation of the compound of the present invention as an oily or aqueous suspension in almost the same manner as with injection The semi-solid composition is preferably an aqueous or oily gel, or an ointment. pH 8 for example, carbonic acid, phosphoric acid, citric acid, hydrochloric acid, hydroxide sodium), preservatives, (for example, esters of para-oxybenzoic acid, chlorobutanol, benzalkonium chloride) and other additives.
A suppository is produced by the preparation of the compound of the present invention as an oily or aqueous, solid, semi-solid, or liquid composition, by a commonly known method. Useful oily bases for such compositions include glycerides or higher fatty acids (eg cocoa butter, uitepols, produced by Dinamite Nobel Company, Germany), moderate fatty acids (eg, MIGLYOL, produced by Dinamite Nobel Company, Germany), and vegetable oils (for example, sesame oil, soybean oil, cottonseed oil).
Aqueous bases include, for example, polyethylene glycols and propylene glycol. Bases for aqueous gels include, for example, natural gums, cellulose derivatives, vinyl polymers and acrylic acid polymers.
BEST MODE FOR CARRYING OUT THE INVENTION The present invention is hereinafter described in more detail by means of, but not limited to, the following reference examples, examples, preparation examples and experimental examples. 1 H-NMR spectra were determined with tetramethylsilylamine as the internal standard, using the GEMINI 200 (200 MHz) spectrometer (produced by Varian, Ltd.) the LAMBDA 300 (300 MHz) spectrometer (produced by JEOL, Ltd) or the Bruker spectrometer AM500 (500 MHz) (produced by Bruker): all values of d are shown in ppm. Unless specifically indicated otherwise, "%" is by weight. The yield is indicated in% mol / mol.
The symbols used here have the following definitions: s: singlet d: doublet t: triplet dt: doublet triplet m: multiplet br: broad The term "at room temperature" indicates the range of around 15 to 25 ° C, but not is built as strictly limiting.
EXAMPLES Reference Example 1 Production of 2-amino-5-phenylthiophene-3-carboxylic acid ethyl ester To a mixture of ethyl cyanoacetate (6.1 g, 50 mmol), sulfur (1.61 g, 50 mmol), triethylamine (3.5 mL) , 25 mmol), and dimethylformamide (10 mL), phenylacetaldehyde (50% solution in diethyl phthalate, 12.05 g, 50 mmol) was added dropwise with stirring at 45 ° C for 20 minutes. After stirring at 45 ° C for 9 hours, the reaction mixture was concentrated and the residue obtained was extracted with ethyl acetate and dried (MgSO 4), the solvent was distilled under reduced pressure. The residue was chromatographed on silica gel and recrystallized from ether-hexane to give pale yellow tubular crystals (5.55 g, 45%). mp 124.5-125.5 ° c (lit., 123-124 ° C). Elemental analysis for C? 3H? 3N02S C (%) H (%) N (%) Calculated: 63.13; 5.30; 5.66 Found: 62.99; 5.05; 5.63 XH-NMR (300MHz, CDCl3) 6: 1.37 (3H, t, J = 7.1Hz), 4.30 (2H, d, J = 7.1Hz), 5.97 (2H, br), 7.17-7.46 (6H, m) . IR (KBr): 3448, 3320, 1667, 1590, 1549 cm -i Reference Example 2 Production of 2-amino-4-methyl-5- (4-methoxyphenyl) thiophene-3-carboxylic acid ethyl ester A mixture of 4-methoxyphenylacetone (16.5 g, 0.10 mol), ethyl cyanoacetate (12.2 g) , 0.10 mol), ammonium acetate (1.55 g, 20 mmol), acetic acid (4.6 mL, 80 mmol) and benzene (20 mL) was heated for 24 hours, while the water that was produced was removed using a Dean apparatus. -Stark. After cooling, the reaction mixture was concentrated under reduced pressure and the residue was partitioned between dichloromethane and aqueous sodium bicarbonate. Then, the organic layer was washed with saline and dried (MgSO 4), the solvent was distilled under reduced pressure. To a solution in methanol (30 mL) of the obtained residue, sulfur was added (3.21 g, 0.10 mol) and diethylane (10.44 mL, 10.10 mol), followed by stirring at 50 to 60 ° C for 2 hours, successively the reaction mixture was concentrated. The residue obtained was extracted with ethyl acetate, washed with saline and dried (MgSO), the solvent was distilled under reduced pressure. The residue was chromatographed on silica gel and recrystallized from ether-hexane to give light yellow tubular crystals (11.5 g, 40%). mp 79-80 ° C. Elemental analysis for C15H17NO3S C (%) H (%) N (%) S (%) Calculated: 61.83; 5.88; 4.81; 11.01 Found; 6X81; 5.75; 4.74; 10.82 XH-NMR (200MHz, CDCI3) d: 1.37 (3H, t, J = 7.1Hz), 2.28 (3H, s), 3.83 (3H, s), 4.31 (2H, q, J = 7.1Hz), 6.05 (2H, brs), 6.91 (2H, d, J = 8.8Hz), 7.27 (2H, d, J = 8.8Hz). IR (KBr): 3426, 3328, 1651, 1586, 1550, 1505, 1485 can "1. FAB-MS m / z: 291 (M +).
Reference Example 3 Using some acetone derivatives in place of 4-methoxyphenylacetone, the following compound was obtained in the same manner as in Reference Example 2.
Yield: 40% mp 64-65 ° C.
Reference Example 4 Production of [3-ethoxycarbonyl-5- (4-methoxyphenyl) -4-methylthiophen-2-yl] aminomethylene-malonic acid diethyl ester To the compound obtained in Reference Example 2 (10 g, 34.3 mmol), diethyl ether of ethoxymethylenemalonic acid (7.45 g, 34.5 mmol) was added, followed by stirring at 120 ° C for 2 hours. After the reaction mixture was cooled, the precipitated crystals were collected by filtration via ether addition, washed again with ether, and dried over phosphorus pentaoxide, under reduced pressure, to give yellow crystals (14.2 g, 90 %). mp 122-123 ° C. -? - NMR (200MHz, CDC13) d: 1.32 (3H, t, J = 7.1Hz), 1.38 (3H, t, J = 7.2Hz), 1.41 (3H, t, J = 7.2Hz), 2.34 (3H , s), 3.85 (3H, s), 4.25 (2H, q, J = 7.1Hz), 4.38 (2H, q, J = 7.2Hz), 4.45 (2H, q, J = 7.2Hz), 6.95 (2H , d, J = 8.8Hz), 7.31 (2H, d, J = 8.8Hz), 8.22 (1H, d, J = 13.4Hz), 12.74 (1H, d, J = 13.1Hz). IR (KBr): 2984, 1720, 1707, 1688, 1653, 1599, 1518, 1499 c '-1 Reference Example 5 Using the compound obtained in Reference Example 3 as starting material, the following compound was obtained in the same manner as in Reference Example 4.
Yield 92% Mp 108-109 ° C.
Reference Example 6 Production of 3-carboxy-5- [(4-methoxyphenyl) -4-methylthiophen-2-yl] aminomethylenemalonic acid diethyl ester To a solution in dioxane (20 mL) of the compound obtained in Reference Example 4 (7.0 g, 15.2 mmol), a solution of potassium hydroxide (5.0 g, 75.7 mmol) in ethanol (30 mL) was added with stirring at the temperature between 60 to 70 ° C. After stirring at this temperature for 1 hr, the reaction mixture was allowed to stand at room temperature for 1 hour. To the mixture was added 2 N hydrochloric acid (40 mL, 80 mmol) with ice cooling, the reaction mixture was concentrated under reduced pressure. The yellow precipitate was collected by filtration, washed with cold water-ethanol, and dried over phosphorus pentaoxide under reduced pressure, to give yellow powders (6.1 g, 93%). mp 184-187 ° C. - H-NMR (200MHz, DMSO-d6) d: 1.24 (3H, t, J-7.1Hz), 1.28 (3H, t, J = 7.2Hz), 2.30 (3H, s), 3.80 (3H, s ), 4.15 (2H, q, J = 7.1Hz), 4.24 (2H, q, J = 7.2Hz), 7.03 (2H, d, J = 8.7Hz), 7.37 < 2H, d7 J = 8.7Hz), 8.08 (1H, d, J = 13.6Hz), 12.41 (1H, d, J = 13.6Hz). IR (KBr): 3422, 2980, 1719, 1653, 1607, 1551, 1512 c "1.
Reference Example 7 Using the compound obtained in the Example of Reference 5 as the starting material, the following compound was obtained in the same manner as in Reference Example 6.
Rendimi in t o: 98% mp 187-190 ° C.
Reference Example 8 Production of 4-hydroxy-2- (4-α-ethoxyphenyl) -3-methylthioeno [2, 3-b] pyridine-5-carboxylic acid ethyl ester Small portions of the compound obtained in Reference Example 6 ( 6.0 g, 13.8 mmol) were added with stirring at 120 ° C to the polyphosphoric acid ester (PPE) (90 mL). After stirring at the same temperature for 30 min, the reaction mixture was poured into ice water, extracted with ethyl acetate. The combined extracts were washed with saline, and dried (MgSO 4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel, to give yellow powders (3.65 g, 77%) for a sample for elemental analysis, the obtained powders were recrystallized from ethanol to obtain yellow crystals, mp 162-163 ° C. Elemental analysis for Ci8Hi7N04S C (%) H (%) N (%) S (%) Calculated: 62.96; 4.99; 4.08; 9.34 Found: 62.89; 5.04; 4.01; 9.34 - • -H-NMR (200MHz, CDC13) d: 1.47 (3H, t, J = 7.1Hz), 2.63 (3H, s), 4.87 (3H, s), 4.49 (2H, q, J = 7.1Hz ), 6.99 (2H, d, J = 8.8Hz), 7.44 (2H, d, J = 8.8Hz), 8.84 (lH, s) r 12.11 (1H, s). IR (KBr): 3434, 2992, 1692, 1601, 1582, 1535, 1504 cm "1. FAB-MS m / z: 344 (MH +).
Reference Example 9 Using the compound obtained in Reference Example 7 as an initial material, the following compound was obtained in the same manner as in Reference Example 8.
Yield: 60% Mp 155-157 ° C.
Reference Example 10 Production of 4-hydroxy-2- (4-nitrophenyl) -3-methylthieno [2, 3-b] pyridine-5-carboxylic acid ethyl ester To a solution in concentrated sulfuric acid (10 mL) of the compound obtained in Reference Example 9 (3.76 g, 12.0 mmol) was added dropwise a solution of sodium nitrate (1.27 g, 15.0 mmol) in concentrated sulfuric acid (5 mL) with ice cooling. After stirring at this temperature for 30 minutes, the reaction mixture was poured into ice water and extracted with chloroform. The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The obtained residue was subjected to chromatography on silica gel, to obtain yellow powders, which were recrystallized from ethanol, to obtain yellow crystals (1.75 g, 41%) mp 260-261 ° C. Elemental analysis for Ci7Hi4N205S C (%) H (%) N (%) Calculated: 56.98; 3.94; 7.82 Found: 56.66; 3.91; 7.86 -? - NMR {200MHz, CDC13) d: 1.49 (3H, t, J = 7.1Hz), 2.70 (3H, s), 4.51 (2H, q, J = 7.1Hz), 7.70 (2H, d, J = 8.8Hz), 8.3 (2H, d, J = 8.8Hz), 8.89 (1H, s), 12.27 (1H, s). IR (KBr): 3002, 1692, 1605, 1514, 1350, 1290 cm "1. FAB-MS m / z: 358 (MH +).
Reference Example 11 Production of 4,7-dihydro-7- (2-methoxybenzyl) -2- (4-methoxyphenyl) -3-methyl-4-oxothieno [2, 3-b] pyridin-5-ethyl ester carboxylic acid To a suspension of sodium hydride (60% dispersion in oil;, 123 mg, 3.08 mmol) in dimethylformamide (3 mL) was added a solution of the compound obtained in the Reference Example 8 (1.0 g, 2.91 mol) in dimethylformamide (20 mL) under a stream of nitrogen with ice cooling. After stirring at this temperature for 30 minutes, a solution of 2-methoxybenzyl chloride (0.92 g, 5.87 mmol) in dimethylformamide (3 mL) was added dropwise to the mixture. After stirring at room temperature for 23 hours and at 70 ° C for 2 hours, the reaction mixture was concentrated, and then the obtained residue was partitioned between ethyl acetate and an aqueous solution of ammonium chloride. The aqueous layer was extracted with ethyl acetate. The combined extracts were washed with saline, and dried (MgSO 4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give yellow amorphous powders (0.95 g, 70%). For a sample for elemental analysis, the obtained powders were recrystallized from dichloromethane-ether to obtain yellow prismatic crystals. mp 165-167 ° C. Elemental analysis for: C26H25NO5S 0.5H2O C (%) H (%) N (%) Calculated: 66.08; 5.55; 2.96 Found: 66.33; 5.44; 2.74 Trl-NMR (200MHz, CDCI3) d: 1.41 (3H, t, J = 7.1Hz), 2.65 (3H, s), 3.85 (3H, s), 3.86, (3H, s), 4.39 (2H, q , J = 7.1Hz), 5.16 (2H, s), 6.92-7.00 (4H,), 7.21-7.41 (4H, m), 8.41 (1H, s). GO . { KBr): 2980, 1727, 1684, 1609, 1590, 1497, 1464 c "1.
Reference Example 12 The following compound was produced by the same method as described in Reference Example 11, using benzyl 2,6-difluorochloride in place of 2-methoxybenzyl chloride with the compound obtained in Reference Example 10 as the initial material.
Yield 81% mp 215-217 ° C.
Reference Example 13 Production of 3-bromoacetyl-4,7-dihydro-7- (2-methoxybenzyl) -2- (4-methoxyphenyl) -4-oxothieno [2, 3-b] pyridin-5-ethyl ester carboxylic A mixture of the compound obtained in Reference Example 11 (0.35 g, 0.755 mol), N-bromosuccinimide (0.135 g, 0.758 mmol), a, a'-azobisisobutyronitrile (13 mg, 0.079 mmol), and carbon tetrachloride ( 5 mL) was heated and refluxed for 2 hours. After cooling, the insoluble substances were filtered, then the filtrate was diluted with chloroform. After, the organic layers were washed with saline and dried (MgSO 4), the solvent was distilled under reduced pressure. The residue obtained was recrystallized from ethyl acetate to obtain colorless needles (0.272 g, 66%). mp 200-201 ° C. Elemental analysis for CaeH ^ NOsSBr C (%) H (%) N (%) Calculated: 57.57; 4.46; 2.58 Found: 57.75; 4.31; 2.31 1H-HMR (200MHz, CDC13) d: 1.40 (3H, t, J = 7.1Hz), 3.86 (6H, s), 4.40. { 2H, q, J = 7.1Hz), 5.05 (2H, s), 5.16 (2H, s), 6.92-7.04 (4H,), 7.23-7.28 (lH,), 7.34-7.43 (1H, m), 7.57 . { 2H, d, J = 8.9Hz), 8.46 (1H, s). IR (KBr): 2980, 1725, 1607, 1588, 1497 cm "1.
Reference Example 14 Using the compounds obtained in the Examples of Reference 12, 22 and 27 as initial materials, respectively, the following compounds were obtained (Reference Examples 14-1 to 14-3), in the same manner as in Reference Example 13.
Reference Example 14-1: Rendimi ent o: 81% Pí 200-202 ° c.
Reference Example 14-2: Yield: 80% Amorphous Reference Example 14-3 Yield: 79% Mw 189-192 ° C.
Reference Example 15 Production of 3- ± ethyl acid ester > Cilaminomethyl-4, 7-dihydro-7-. { 2-methoxybenzyl) -2- (4-methoxyphenyl) -4-oxothieno [2,3-b] pyridine-5-carboxylic acid To a solution of the compound obtained in Reference Example 13 (0.245 g, 0.452 mmol) in dimethylformamide ( 5 mL) triethylamine (0.10 mL) was added., 0.717 mmol) and benzylamine (80 μl, 0.732 mmol) with ice cooling. After stirring at room temperature for 1.5 hours, the reaction mixture was concentrated, and then the residue obtained was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous layer was extracted with ethyl acetate. The combined organic extracts were dried (MgSO), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to obtain a colorless oil (0.135 g, 53%). To a solution of this oil in ethanol (4 mL) was added ethanolic hydrochloric acid ION (0.2 mL) with ice cooling, followed by stirring at this temperature for 10 minutes. The reaction mixture was concentrated, and then, the obtained residue was crystallized from ethyl acetate-ether to obtain the hydrochloride (0.113 g) as white crystals. pf [Hydrochloride] 118-119 ° C. Elemental analysis for C33H32N205S HCl 0.9H20 C (%) H (%) N (%) Calculated: 63.79; 5.64; 4.51 Found: 64.03; 5.44; 4.51 - H-NMR. { 300MHz, CDC13) [Free amine] d: 1.40. { 3H, t, J = 7.1Hz), 2.05 (1H, br), 3.81 (3H, s), 3.86 (3H, s), 3.87 (2H, s), 3.94 (2H, s), 4.40 (2H, q , J = 7.1Hz), 5.18 (2H, s), 6.80 (2H, d, J = 8.8Hz), 6.91-6.99 (2H, m), 7.20-7.42 (9H, m), 8.45 (1H, s) . IR (KBr) [Hydrochloride]: 3422.2938, 1719.1605, 1560.1545, Reference Example 16 Production of 4-methyl-2- [(4-methyl-3-oxo-l-penten-1-yl) amino] -5-phenylthiophene-3-carboxylic acid ethyl ester The compound obtained in Example of Reference 3 (10 g), l-methoxy-4-methyl-l-penten-3-one with 85% content (6.9 g), p-toluenesulfonic acid monohydrate (0.219 g), and toluene (100 mL) they were mixed and stirred at room temperature for 2.5 hours. Then, the reaction mixture was diluted with ethyl acetate and washed with saturated aqueous sodium hydrogen carbonate solution, the aqueous layer was extracted with ethyl acetate. The combined extracts were washed with saturated saline, the organic layers were dried over anhydrous magnesium sulfate. The residue was crystallized via the addition of a seed crystal, after which it was triturated with hexane, collected by filtration, and washed, to give the title compound (12.64 g, 92%), mp 104-108 °. C.
Reference Example 17 Production of 4,7-dihydro-7- (2-methoxybenzyl) -2- (4-methoxyphenyl) -3-methyl-4-oxothieno [2,3-b] pyridine-N-benzylpiperazinyl-5- carboxamide A 1-benzylpiperazine (0.77 g, 4.37 mmol) was added dropwise a solution of diisobutyl aluminum hydride (DIBAL) in toluene (1.5 N, 2.9 mL, 4.37 mmol) with ice cooling. After the addition was complete, the solution was allowed to warm to room temperature and was stirred for another 0.5 hour.To this solution was added a solution of the compound obtained in Reference Example 11 (0.50 g, 1.08 mmol) in toluene (5 mL) at room temperature, then the mixture was stirred again at room temperature for 15 hours, methylene chloride 830 mL was added) After washing with water, the solution was dried over sodium sulfate, and then the solvent was concentrated under reduced pressure to give a solid (1.03 g), which was recrystallized from methylene chloride-n-hexane, to give the title compound. {0.48 g, 78%) .pf 233-235 ° C Elemental analysis for C35H35N3O4S 1 / 2H20 C {%) H (%) N (%) S (%) Calculated: 69.75; 6.02: 6.97; 5.32. Found: 69.88; 6.06: 6.98; 5.39 .1H-NMR (300MHz, CDCI3) d: 2.45-2.55 (4H, m), 2.63 (3H, s), 3.43-3.45 {2H, m), 3.55 (2H, s), 3.73-3.82 (2H ,), 3.84 (6 H, s), 5.11 (2H, s), 6.89-6.98 (4H, m), 7.21-7.40 (9H, m), 7.79 (1H, s).
Reference Example 18 Using the compound obtained in Reference Example 12 as starting material and N, 0-dimethylhydroxy, instead of 1-benzylpiperazine, the following compound was obtained in the same manner as in Reference Example 17.
Yield: 80% Pf 223-224 ° C.
Reference Example 19 Production of 4-hydroxy-5-isobutyryl-3-methyl-2-phenyltiene [2, 3-b] pyridine The compound obtained in Reference Example 16 (50 g) and diphenylether (500 mL) was they mixed, and then, this mixture was heated and refluxed for 4 hours, while the methanol that was produced with the progress of the reaction was distilled. After cooling, the diphenylether was distilled under reduced pressure, and then the precipitated crude crystals were washed with n-hexane to give the title compound. { 35.1 g, 81%). Mp 114-117 ° C.
Reference Example 20 Using the compound obtained in Reference Example 14-1 as starting material and N-methoxybenzylamine instead of benzylamine, the following compounds were obtained (Reference Examples 20-1 to 20-3), in the same manner as in Reference Example 15.
Reference Example 20-1 Ren i mi ent o: 83% Pf 140-144 ° C.
Reference Example 20-2; Yield 91% Mp 145-147 ° C.
Reference Example 20-3: Yield: 78% mp 175-177 ° C.
Reference Example 21 Using the compounds obtained in Reference Example 20 as the starting materials (Reference Examples 21-1 to 21-3), were obtained in the same manner as in Reference Example 28.
Reference Example 21-1: Yield: 79% mp 158-160 ° C.
Reference Example 21-2: Yield: 96% mp 195-196 ° C.
Reference Example 21-3: Yield :: 71% mp 144-146 ° C.
Reference Example 22 Production of 7- (2,6-difluorobenzyl) -4,7-dihydro-5-isobutyl-3-methyl-4-oxo-2-phenyl-thieno [2,3-b] pyridine The compound obtained in Reference Example 19 (35 g), potassium carbonate (18.6 g), and N, N-dimethylformamide (280 mL), were mixed, and then 2,6-difluorobenzyl bromide (27.9 g) was added, followed by by stirring at room temperature for 4 hours. Water was added dropwise to the reaction mixture. { 560 mL), followed by stirring for 30 minutes, and then the mixture was stirred for 1 hour with ice cooling. The crude crystals were collected by filtration, washed with water, and dried, successively suspended in a 1: 1 solvent mixture. { 250 mL) of ethyl acetate or diisopropyl ether and stirred at 25 to 40 ° C for 1 hour, then stirred with ice cooling for 1 hour, the residual crystals were collected by filtration and washed with the solvent mixture. from above (125 mL) to give the title compound (44.6 g, 92%). mp 205-207 ° C.
Reference Example 23 Using the compounds obtained in Reference Example 14-2 and Reference Example 24 as the starting materials and N-methylbenzylamine, instead of benzylamine, the following compounds were obtained (Reference Examples 23-1 to 23 -2), in the same manner as in Reference Example 15.
Reference Example 23-1: Yield: 83% mp 197-199 ° C.
Reference Example 23-2 Retreatment: 66% mp 151-152 ° C.
Reference Example 24 * Production of 3-bromomethyl-7- (2,6-difluorobenzyl) -4,7-dihydro-5-isobutyryl-2- (4-nitrophenyl) -4-oxothieno [2,3-b] pyridine After the compound obtained in the Example of Reference 14-3 (1 g), dissolved in methanesulfonic acid (5 mL) with a cooling of 10 to 12 ° C, a solution of sodium nitrate (0.165 g) in methanesulfonic acid (2.5 mL) was added dropwise to this mixture. After being stirred for 2 hours, the mixture was poured into cold water, and then the crystalline precipitate was collected by filtration, washed with water and diisopropyl ether, and dried to give a crude product (1.04 g), which it was then triturated with ethyl acetate (15 mL), then cooled with ice, filtered, and washed with cold ethyl acetate to give the title compound (0.647 g, 59%). Mp 202 to 204 ° C (recrystallized from methanol).
Reference Example 25 Production of 4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-nitrophenyl) -3- (N-benzyl-N-methylaminomethyl) -4-oxothiene [2, 3- b} j > iridine-5- (N-isopropyl) carboxamide To a solution of isopropylamine (0.296 g, 5 mmol) in anhydrous methylene chloride (5 mL) was added dropwise a solution of trimethylaluminum in hexane (15%, 2.41 mL, 5.0 mmol) at 0 ° C. After the addition was complete, the solution was allowed to warm to room temperature and further stirred for 1 hr. To this solution was added a solution of the compound obtained in Reference Example 20-2 (0.12 g, 0.25 mmol) in anhydrous methylene chloride (3 mL) with ice cooling (0 ° C) for 30 minutes. Then, the mixture was stirred at room temperature for another hour, chloroform was added, followed by washing with water. The combined organic layers were dried with sodium sulfate, and then the solvent was concentrated under reduced pressure, to give a solid, which was recrystallized from chloroformyl acetate-ethyl ether, to give colorless crystals (0.96 g, 70% ). mp 200-202 ° C. 1H-NMR (500MHz, CDC13) [Free amine] d: 1.30 (6H, d, J = 6.7Hz), 2.15 (3H, s), 3.66 (2H, s), 4.18 (2H, s), 4.18-4.31 (1H, m), 5.32 (2H, s), 7.00 (2H, t, J = 7.26Hz), 7.13-7.25 (5H, m), 7.42 (1H, t, J = 7.3Hz), 8.02 (2H, d, J = 8.9Hz), 8.26 (2H, d, J = 8 .9Hz), 8.73 (1H, s), 10.02 (1H, d, J = 9.1Hz). IR (KBr): 2974, 1661, 1597, 1547, 1497, 1346, 1212, 1035 cm "1 FAB-Mass m / z 617 (MH) +.
Reference Example 26 Using the compound obtained in Reference Example 20-2 as an initial material and N, 0-dimethylhydroxyamine, the following compound was obtained, in the same manner as in Reference Example 25.
Yield: 96% mp 100-102 ° C.
Reference Example 27 Production of 4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-nitrophenyl) -5-benzyl-3-methyl-4-oxothieno [2,3-b] pyridine The The compound obtained in Reference Example 18 (3.93 g, 7.87 mmol) was dissolved in anhydrous tetrahydrofuran (THF) with slight heating. While this solution was maintained at 0 ° C, a solution of phenylmagnesium bromide in THF (1M, 15.7 mL, 15.7 mmol) was added dropwise to the mixture over 10 minutes. After the addition was complete, the solution was stirred for an additional 1 hour. The reaction mixture was partitioned between ethyl acetate (200 mL) and water. { 50 L), and then the aqueous layer was extracted again with ethyl acetate. The combined organic layers were dried with magnesium sulfate, the solvent was concentrated under reduced pressure, to give a residue, which was chromatographed on silica gel, to give yellow crystals (3.00 g, 74%), which they were recrystallized from ethyl acetate-hexane, to give yellow crystals. mp 114-116 ° C. Elemental analysis for C28H? 8N204SF2 0.7H20 C (%) H (%) N (%) Calculated: 63.56; 3.70; 5.29 Found: 63.83; 3.95; 5.08 H-NMR (500MHz, CDC13) d: 2.68 (3H, s), 5.30 (2H, s), 7.02 (2H, t, J = 8.lHz), 7.43 (3H, t, J = 7.2Hz), 7.52-7.63 (3H, m), 7.86. { 2H, d, J = 7.5Hz), 7.99 (1H, s), 8.30 (2H, d, J = 8.7Hz). IR (KBr): 3422, 3068, 1665, 1615, 1491, 1473, 1346, 853 cm "1. FAB-Mass m / z 517 (MH) +.
Reference Example 28 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-aminophenyl) -5-benzoyl-4-oxothiene [ 2, 3-b] pyridine To a mixture of the compound obtained in Reference Example 23-1 (0.30 g, 0.47 mmol), in ethyl alcohol (6 mL), water (2 mL) was added, and then a drop of concentrated hydrochloric acid was added to give a uniform solution. - To this solution were added iron powder. (0.105 g, 2.0 mmol) and concentrated hydrochloric acid (0.39 mL, 4.7 mmol) carefully. After the addition was complete, the mixture was stirred at room temperature for 5 hours and filtered through Celite. A small amount of aqueous ammonia was added, and then, the reaction filtrate was concentrated under reduced pressure. The obtained residue was poured into ice water, neutralized with sodium bicarbonate, and extracted with ethyl acetate. After, the combined organic layers were washed with saline, and dried (MgSO4), the solvent was distilled off under reduced pressure. The residue obtained was subjected to chromatography on silica gel and recrystallized from isopropyl ether, to give yellow needles (0.24 g, 84%). mp 126-128 ° C. Elemental analysis for C36H29N302SF2 1 / 2H20 C (%) H (%) N (%) Calculated: 68.93; 5.04; 6.70 Found: 68.71; 5.18; 6.62 - H-EuMR. { 300MHz, CDC13) d: 2.13 (3H, s), 3.65 (2H, s), 3.87 (2H, br s), 4.14 (2H, s), 5.28 (2H, s), 6.74 (2H, d, J = 8.7Hz), 7.00. { 2H, t, J = 7.8Hz), 7.16-7.24 (5H, m), 7.36-7.46 (3H, m), 7.53 (1H, t, J = 7.2Hz), 7.62 (2H, d, J = 8.4Hz ), 7.89 (2H, d, J = 7.2Hz), 7.94 (1H, s). IR (KBr): 3358, 1607, 1495, 1473, 1035 cm "1. FAB-Mass m / z 606 (MH) +.
Reference Example 29 Production of 2- (4-aminophenyl) -7-. { 2,6-difluorobenzyl) -4,7-dihydro-5-isobutyryl-3- (N-benzyl-N-methylaminomethyl) -4-oxothieno [2,3-b] pyridine To a solution of the compound obtained in Reference Example 23-2 (0.25 g, 0.415 mmol) in methanol (5 mL) were added dropwise (0.093 g, 1.66 mmol) and concentrated hydrochloric acid. { 0.8 mL) with ice cooling. After the addition was complete, the mixture was stirred at room temperature for 1 hour and filtered through Ceuta. A small amount of aqueous sodium hydrogen carbonate solution was added, followed by extraction with methylene chloride (30 mL x 3). The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give light yellow amorphous powders (0.203 g, 86%).
^ -NMR (300MHz, CDC13) d: 1.18 (6H, d), 2.11 (3H, s), 3.65 (2H, s), 3. 85 { 2H, br s), 4.17 (2H, s), 4.18 (1H, m), 5.25 (2H, s), 6. 73 (2H, d), 6.95 (2H, t), 7.10-7.26 (5H,), 7.42 (1H, m), 7.58. { 2H, d), 8.27. { 1H, s).
Reference Example 30 Production of 5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-3- (N-benzyl-N-methylaminomethyl) -2- (4-propionylaminophenyl) -4-oxothiene [ 2, 3-b] pyridine The compound obtained in Reference Example 28 (0.14 g, 0.23 mol) was dissolved in anhydrous methylene chloride (2 mL), and then triethylamine (0.038 L) was added to the mixture with cooling of ice (0 ° C). After this solution was stirred for a moment, propionyl chloride (0.021 mL, 0.243 mmol) was added to the mixture, after the addition was complete, the solution was further stirred with ice cooling (0 ° C) for 40 minutes. minutes The reaction mixture was partitioned between methylene chloride (25 mL) and barely covered with a layer of water (10 mL). The aqueous layer was extracted again with methylene chloride (25 mL). The combined organic layers were washed with saline, and dried (MgSO 4), the solvent was distilled under reduced pressure. The solid obtained was recrystallized from ethyl acetate-isopropyl ether, to give yellow needles (0.10 g, 65%). mp 226-228 ° C. Elemental analysis for C39H33N303SF2 0.7H20 C (%) H (%) N (%) Calculated: 69.46; 5.14; 6.23 Found: 69.60; 5.18; 6.04 The compound thus obtained was dissolved in ethyl acetate, and then a saturated solution of hydrogen chloride (HCl) in ether was added, in an amount equal to slightly in excess, and then the crystalline precipitate was recrystallized from ether isopropyl, to give light yellow needles (0.095 g, 61%). mp 218-220 ° C. Elemental analysis for C39H33N3O3SF2 HCl 3.5H20 C (%) H (%) N (%) Calculated: 61.53; 5.43; 5.52 Found: 61.83; 5.33; 5.30 1H-HMR (300MHz, DMSO-d6) d: 1.11 (3H, t, J = 7.2Hz), 1.93 (3H, s), 2.35 (2H, q, J = 7.5Hz), 3.44 (2H, s) , 4.00 (2H, s), 5.62 (2H, s), 7.11-7.25 (6H, m), 7.43-7.72 (10H, m), 7.79. { 2H, d, J = 7.5Hz), 8.40 (1H, s), 10.03 (1H, s). IR (KBr): 3422, 3068, 1603, 1502, 1473, 1035 c "1. FAB-Mass m / z 662 (MH) +.
Reference Example 31 The following compounds (Reference Example 31-1 to 31-10) were produced by the same method as that described in Reference Example 30, using various chlorides of acid, isocyanates, and esters of chlorocarbonic acid, in place of propionyl chloride, and, also using the amines shown in Example 16 and carbonyldiimidazole, with the compounds obtained in Reference Examples 28 and 29 as the starting materials. The following compounds (Reference Examples 31-11 to 31-16) can be produced in the same manner.
Reference Example 31-1-1: Yield: 68% Mp 238-240 ° C.
Reference Example 31-1-2: mp 230-232 ° C.
Reference Example 31-2-1: yield: 64% mp 201-204 ° C.
Reference Example 31-2-2: Reference Example 31-3-1: Yield: 55% Mw 207-210 ° C.
Reference Example 31-3-2: mp 222-226 ° C, Reference Example 31-4: Yield: 49% mp 185-187 ° C Reference Example 31-5: Yield: 79% Mp 216-218 ° C.
Reference Example 31-6; Yield: 73% Mp 180-183 ° C.
Reference Example 31-7 Yield: 65% Mw 245-247 ° C.
Reference Example 31-8: Performance 65% Reference Example 31-9: Rendimi at t o: 70% Mw 232-234 ° C.
Reference Example 31-10: Yield: 73% Mw 192-197 ° C.
Reference Example 31-11; Reference Example 31-12: Reference Example 31-13: Reference Example 31-14 Reference Example 31-15: Reference Example 31-16: Reference Example 32 Production of 3- (-N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-nitrophenyl) -5-benzoyl-4-oxothiene [2, 3-b] pyridine The compound obtained in Reference Example 26. { 1.91 g, 3.09 mmol) was dissolved in anhydrous tetrahydrofuran (THF) (30 mL), with heating, and then added to the. drop by drop, a solution of phenylmagnesium bromide in THF (1M, 6.18 mL, 6.2 mmol) with ice cooling (0 ° C) for 10 minutes. After stirring under the conditions of ice cooling for 1 hr, the reaction mixture was partitioned between ethyl acetate (100 mL) and aqueous hydrochloric acid (0.5N, 100 mL), and then, the organic layer was washed again once with saturated saline solution (10 mL). After the organic layer was dried (MgSO), the solvent was distilled under reduced pressure. The obtained residue was subjected to chromatography on silica gel, to give yellow crystals (1.00 g, 51%), which were then recrystallized from isopropyl ether, to give yellow needles. mp 197-199 ° C. Elemental analysis for C36H27N304SF2 0.7H20 C (%) H (%) N (%) Calculated: 66.70; Four . 42; 6.48 Found: 66. 59; Four . 48; 6. 42 1 H-NMR (300MHz, CDCl 3) d: 2.17 (3H, s), 3.61 (2H, s), 4.16 (2H, s), 5.30 (2H, s), 7.03 (2H, t, J; 8.1 Hz), 7.19-7.25. { 5H,), 7.40-7.47 (3H, m), 7.56 (1H, t, J = 7.5Hz), 7.88 (2H, d, J = 6.9Hz), 7.96 (1H, s), 8.10 (2H, d, J = 8.7Hz), 8.28 (2H, d, J = 8.7Hz). IR (KBr): 3430.1663, 1611, 1518, 1473, 1348, 853 c "1. FAB-Mass m / z 636 (MH) +.
Reference Example 33 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-isobutyrylaminophenyl) -5- (2-bromoisobutyryl) - 4-Oxothieno [2, 3-b] pyridine The compound obtained in Reference Example 31-10 (0.48 g, 0.75 mmol) was dissolved in dichloromethane (15 mL). To this solution was added dropwise 47% hydrobromic acid 80.35 L, 3.0 mmol) at room temperature for 1 minute, followed by further stirring for 10 minutes. To this reaction mixture was added dropwise bromine (39 μl, 0.75 mmol), followed by further stirring at room temperature for 24 hours. The reaction mixture was partitioned between dichloromethane (30 mL) and aqueous sodium bicarbonate (saturated, 60 mL), and then the organic layers were washed again with saturated saline (100 mL). After the organic layer was dried (MgSO 4) the solvent was distilled under reduced pressure to give brown amorphous crystals (0.53 g).
Reference Example 34 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-isobutyrylaminophenyl) -5-methacryloyl-4-oxothiene [ 2, 3-b] pyridine The compound obtained in Reference Example 33. { 0.52 g, 0.72 mmol) was dissolved in dimethylformamide (DMF) (30 mL), and then, to this solution was added potassium acetate (2.0 g, 20 mmol) at room temperature, followed by stirring at 100 ° C for 20 minutes. hours. The reaction mixture was partitioned between ethyl acetate (30 mL) and water (30 mL), and then, the aqueous layer was washed again with ethyl acetate (10 mL). The combined organic layers were washed again with saturated saline (30 mL). After, the organic layer was dried (MgSO), the solvent was distilled under reduced pressure to give a brown oil (0.56 g).
Example 1 Example 1-1: Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4 -oxotiene [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (2.57 g, 4.5 mmol) in dichloromethane (30 mL) were added cyclopropanecarboxylic acid (861 mg, 10 mmol) and diisopropylethylamine (2.59 g, 20 mmol), and then benzotriazole acid -1-yl-oxy triprrolidino phosphonium hexafluorophosphoric acid (PyBop) (5.40 g, 10 mmol), was added to it in a small portion with ice cooling. After stirring at room temperature for 1 day, the reaction mixture was poured into a 0.1 N aqueous solution of potassium hydroxide and extracted with chloroform. The combined extracts were washed with saline, and dried (Na2SO), the solvent was distilled under reduced pressure. The obtained residue was chromatographed on silica gel and recrystallized from ethyl acetate to give white powdered crystals (2.15 g, 79%). 1 H-NMR (300MHz, CDCl 3) d: 0.85-0.90 (2H, m), 1.10-1.15 (2H, m), 1.18 (6H, d), 1.53-1.59. { 1H, m), 2.09 (3H, s), 4.12-4.20 (1H, m), 4.16. { 2H, s), 5.25 (2H, s), 6.99. { 2H, t), 7.10-7.25 (6H, m), 7.35-7.45 (lH, m), 7.61 (2H, d), 7.67 (1H, br s), 7.76. { 2H, d), 8. 27 (1H, s). FAB-Mass m / z 608 (MH) +.
JExample 1-2: Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4 hydrochloride -oxotiene [2, 3-b] pyridine The white powdered crystals obtained in Example 1-1 were dissolved in ethanol, and then, a solution of hydrogen chloride in ethyl ether was added in a small amount in excess, followed by recrystallization from ethanol-ethyl ether. , to give powdered crystals of white color (2.20 g, 72%). mp 254-256 ° C. Elemental analysis for C37H35N303SF2 HCl C. { %) H (%) N (%) Calculated: 65.72; 5.37; 6.21 Found: 65.61; 5.38; 6.25 IR (KBr): 2950, 1673, 1595, 1502, 1473, 1408, 1313, 1183 cm "1.
Example 2 Production of 4,7-dihydro-2- [4- (3-acetoxy-2-methylpropionylamino) phenyl] -7- (2,6-dicluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5 -isobutyryl-4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 20 (1.14 g, 2.0 mmol) in dichloromethane (30 mL) were added dropwise, triethylamine (500 mg, 5 mmol) and 3-acetoxyisobutyryl chloride (0.5 g, 3 g). mmol) with ice cooling for one minute, followed by further stirring at room temperature for 1 hr. The reaction mixture was partitioned between an aqueous solution of sodium hydrogen carbonate (30 mL) and dichloromethane (30 mL), and then the organic layer was extracted again with dichloromethane (10 mL). The combined extracts were washed with saline, and dried (Na2SO4), the solvent was distilled off under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give brown crystals. { 1.4 g). XH-NMR (300MHz, CDC13) d: 1.18 (6H, d), 1.30 (3H, d), 2.08 (3H, s), 2.12 < 3H, s), 2.73-2.84. { lH, m), 3.65 (2H, s), 4.11-4.36 (3H, m), 4.17 (2H, s), 5.27 (2H, s), 7.00 (2H, t), 7.13-7.22 (5H, m) , 7.36-7.46. { lH, m), 7.52 (lH, br s), 7.62-7.82 (4H, q), 8.29 (1H, s).
EXAMPLE 3 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -4-oxothieno [2, 3-b] pyridine the compound obtained in Example 2 (1.40 g) was dissolved in methanol (100 mL), and then potassium carbonate (2.0 g, 15 mmol) was added to the solution, followed by stirring at room temperature for 15 minutes. The reaction mixture was partitioned between ethyl acetate (200 mL) and distilled water (300 mL), and then the aqueous layer was extracted with ethyl acetate (100 mL). The combined extracts were washed with saline, and dried (Na2SO4), the solvent was distilled off under reduced pressure. The residue obtained was subjected to chromatography on silica gel from ether to give white crystals (0.95 g, 72%). mp 161-162 ° C. Elemental analysis for C37H3N30SF2 0.5H20 C (%) H (%) N (%) Calculated: 66.65; 5.74; 6.30 Found: 66.70; 5.52; 6.40 -? - NMR (300MHz, CDC13) d: 1.17 (6H, d), 1.18 (3H, d), 2.06 (3H, s), 2.67-2.78. { lH, m), 3.61 (2H, s), 3.57-3.84 (2H, m), 3.99- .08 (1H, m), 4.15 (2H, s), 5.52 (2H, s), 7.08 (2H, t ), 7.15 (5H, s), 7.44-7.54 (lH, m), 7.58. { 2H, d), 7.69 (2H, d), 8.46 (1H, s). FAB-Mass m / z 658 (MH) +.
Example 4 Using the compound obtained in Reference Example 29, the following compounds were obtained (Examples 4-1 to 4-3), in the same manner as in Example Example 4-1: Yield: 67% Pf 201-203 ° C Example 4-2: Yield: 58% Mp 135-137 ° C.
Example 4-3: Performance: 75 'Pf 205-207 ° C Example 5 Example 5-1: Production of 3-. { N-benzyl-N-methylaminomethyl) -4,1-dihydro-7- (2,6-difluorobenzyl) -2- [4- (2-hydroxyacetylamino) phenyl] -5-isobutyryl-4-oxothiene [2, 3- b] pyridine The compound obtained in Example 4-1 (0.137 g) was dissolved in dichloromethane (1.0 mL), then, to 1 solution, trifluoroacetic acid (1.0 mL) was added dropwise with ice cooling. After stirring under the conditions of ice cooling for 1 hour, the reaction mixture was concentrated to dryness under reduced pressure, and then, the obtained residue was fractionated between chloroform (50 mL) and saturated aqueous sodium bicarbonate (50 mL). ), and then, the aqueous layer was extracted again with chloroform (10 mL). The combined organic layers were washed with saturated saline (30 mL). Then, the organic layer was dried (MgSO4), the solvent was distilled off under reduced pressure, and then, the obtained residue was chromatographed on silica gel and recrystallized from ethanol-ethyl acetate to give white crystalline powders ( 0.12 g, 95%).
Example 5-2: Production of 3- hydrochloride. { N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- [4- (2-hydroxyacetylamino) phenyl] -5-isobutyryl-4-oxothiene [2, 3- b] pyridine The white crystalline powders obtained in Example 5-1, were dissolved in dichloromethane (1.0 mL), and then to this solution was added an ethanolic solution containing IN hydrogen chloride (10 mL), with ice cooling. The solvent was distilled under reduced pressure, and then, the residue was recrystallized from ethanol-ethyl acetate to give white crystalline powders (0.12 g, 84%). mp 197-199 ° C. Elemental analysis for C35H33N304SF2 HCl 2. 5H20 C (%) H (%) N (%) Calculated: 59.11; 5.53; 5.91 Found: 59.06; 5.25; 5.93 - H-NMR (300MHz, CDC13) d: 1.18 (6H, d), 2.10 (3H, s) «. 3.63 (2H, s), 3.72 (lH, br), 4.09-4.17 (lH, m), 4.16 (2H, s), 4.24 (2H, s), 5.24 (2H, s), 6.99 (2H, t) , 7.18-7.23 (5H, m), 7.35-7.45 (lH, m), 7.63. { 2H, d), 7.73 (2H, d), 8.25 (lH, s), 8.59 (lH, s).
Example 6 Using the compounds obtained in Example 4-2 and Example 4-3, the following compounds were obtained (Examples 6-1 to 6-2), in the same manner as in Example 5.
Example 6-1: Yield: 76% Mw 192-194 ° C.
Example 6-2: Yield: 81% Mp 193-195 ° C.
Example 7 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- (2,6-difluorobenzyl) -2- (4-isobutyrylaminophenyl) -5- (2-hydroxy-2-methylpropionyl) ) -4-oxothieno [2, 3-b] pyridine Amorphous brown crystals (0.13 g), obtained by repeating the method described in Reference Example 33, using the compound obtained in Reference Example 31-10, were hydrolysed with potassium acetate by the same method as described in Reference Example 34. The solid obtained was subjected to chromatography on silica gel to remove the compound obtained in Reference Example 34, and the by-product was recrystallized from methanol to give colorless crystalline powders (0.03 g). mp 170-172 ° C.
Elemental analysis for C37H37N304SF2 1 / 6H20 C (%) H (%) N (%) Calculated: 67.26; 5.69; 6.36 Found: 67.18; 5.74; 6.46"-H-NMR (300MHz, CDC13) d: 1.28 (6H, d), 1.49 (6H, s), 2.06 (3H, s), 2.51-2.60 (lH, m), 3.61 { 2H, s ), 4.13 (2H, s), 5.31 (2H, s), 7.01 (2H, t), 7.12-7.21 (5H, m), 7.37-7.47 (2H, m), 7.64 (2H, d), 7.74 {2H, d), 8.04 { LH, s), 8.35 { LH, s). IR (KBr): 3466,1669, 1499, 1313, 1075 cm "1.
Example 8 Production of 3-. { N-benzyl-N-methylaminomethyl) -4,7-dihydro-7- [2,6-difluorobenzyl] -2- (4-isobutyrylaminophenyl) -5- (2-hydroxy-2-methylpropionyl) -4-oxothiene [2 , 3-b] pyridine.
The compound obtained in Reference Example 34 (0.56 g) was dissolved in trifluoroacetic acid (10 mL) and stirred at room temperature for 1 hour. The reaction mixture was concentrated to dryness under reduced pressure, and the residue obtained was partitioned between dichloromethane (30 mL) and saturated aqueous sodium bicarbonate (60 mL), and then, the aqueous layer was extracted again with dichloromethane (10 mL). ). The combined organic layers were washed with saturated saline. { 30 mL) i After, the organic layer was dried (MgSO 4), the solvent was distilled under reduced pressure, to give amorphous brown crystals (0.42 g), which were subjected to chromatography on silica gel, to give crystals yellow (0.07 g, total recovery ratio as calculated from the compound obtained in Reference Example 31-10 12%). ^ - MR (300MHz, CDC13) d: 1.15 (3H, d), 1.26 (6H, d), 2.07 (3H, s), 2.53-2.62 (lH, m), 3.58 (2H, d), 3.66 (2H , d), 3.82 (1H, dd), 3.93 (1H, dd), 4.03 (1H, d), 4.22 (1H, d), 4.27-4.37. { 1H, m), 5.26 (2H, s), 6.99 (2H, t), 7.13-7.21 (5H, m), 7.36-7.46 (1H, m), 7.63 (2H, d), 7.67 (2H, d) 7.79 (1H, s), 8.30 (1H, s).
Example 9 Production of (R) -4,7-dihydro-2- [4- (3-t-butoxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl) N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (1.14 g, 2.0 mmol) in dichloromethane (20 mL), diisopropylamine (520 mg, 4 mmol) and (2R) -3-t-butoxy-2-acid were added. methylpropanoic acid (0.43 g, 3 mmol), followed by stirring with ice cooling. To the solution was added benzotriazol-1-yloxytrimethylaminophosphonium hexafluorophosphate (BOP reagent). { 1.33 g, 3 mmol). After the. stirring with ice cooling, for 1 hour, the solution was subsntly stirred at room temperature for 4 hours. The reaction mixture was concentrated to dryness under reduced pressure, and then the obtained residue was partitioned between water (50 mL) and chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel, to give yellow powders (1.11 g, 78%).
XH-NMR (300MHz, CDCl 3) d: 1.18 (6H, d), 1.25 (3H, d), 1.30 (9H, s), 2.62. { 3H, s), 2.63-2.71 (lH, m), 3.51 (1H, similar at), 3.61 (1H, dd), 3.64 (2H, s), 4.18 (2H, s), 4.18 (1H, quintuplete), 5.27. { 2H, s), 7.00 (2H, t), 7.14-7.22 (5H, m), 7.36-7.46 (1H,), 7.60 (2H, d), 7.76 (2H, d), 8.28 (1H, s), 9.00 (lH, s).
Example 10 Example 10-1: Production of (R) -4,7-dihydro-2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N -benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine The compound obtained in Example 9 (1.11 g,) was dissolved in trifluoroacetic acid (5 mL), under the conditions of ice cooling, and then, it was then stirred for 1 hour, the solution was allowed to warm to the temperature environment and stirred for another 12 hours. The reaction mixture was concentrated to dryness under reduced pressure, and then dissolved in methanol (20 mL) and again cooled with ice, and then a 5N solution of sodium hydroxide was added to reach a pH of 9.0. The reaction mixture was stirred for 30 minutes and then stirred at room temperature for 30 minutes. The reaction mixture was concentrated to dryness under reduced pressure at low temperature, and then the residue obtained was subjected to chromatography on silica gel. The yellow amorphous crystals obtained (0.83 g, 82%) were recrystallized from ether to give light yellow powdered crystals (0.69 g). mp 161-162 ° C. Elemental analysis for C37H37N304SF2 0.5H20 C (%) H (%) N (%) Calculated: 66.65; 5.74; 6.30 Found 66.47 5.73 6.10 -? - NMR (300MHz, CDC13) d: 1.18 (6H, d), 1.28 (3H, d), 2.11 (3H, s), 2.71 (lH, m), 3.65 (2H, s) , 3.83-3.85 (2H, m), 4.16 (1H, quint), 4.18 (2H, s), 5.27 (2H, s), 7.O0 (2H, t), 7.14-7.23 (5H, m), 7.36 -7.44. { lH, m), 7.64 (2H, d), 7.77. { 2H, d), 8.09 (1H, s), 8.28 (lH, s). FAB-Mass m / z 658 (MH) + Example 10-2: Production of (R) -4,7-dihydro-2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-) hydrochloride methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine The title compound was obtained in the same manner as in Example 1. Mp 214-216 ° C.
Example 11 Example 11-1: Production of 4,7-dihydro-2- [4- (2-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N -methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (L14 g, 2.0 mmol) in dichloromethane, (30 mL) were added diisopropylethylamine (1.04 g, 8 mmol) and 2-hydroxyisobutanoic acid (0.416 g, 4 mmol), followed by by stirring under the conditions of ice cooling. To this solution was added benzotriazol-1-yloxytrimethylaminophosphonium hexafluorophosphate (BOP reagent) (1.76 g, 4 mmol). After stirring under the conditions of ice cooling for 1 hour, the solution was further stirred at room temperature for 4 days. The reaction mixture was concentrated to dryness under reduced pressure, and then the residue obtained was partitioned between water (50 mL) and chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The residue obtained was chromatographed on silica gel and recrystallized. from ether, to give crystals μilvei zad acerillos (0.56, 43%) Elemental Jólisis for C H N O SF 0.31 0 3? 37 3 4 Calculated C (%) H (%) N (%) 66.65; .74; 6.30 Ercrptrado 66.54; 5.49; 6.36 1H-NMR (300MHz, CDC13) d: 1.58 (6H, s), 2.10 (3H, s), 3. 05 (1H, brs), 3.64 (2H, s), 4.16 (2H, s ), 4.12-4.22 (lH, m), 5.23 (2H, s), 6. 99 (2H, t), 7.05-7.25 (5H, m), 7.34-7.44 (lH, m), 7.64 (2H, d), 7. 75 (2H, d), 8.25 (1H, s), 8.86 (1H, s).
Example 11-2: Production of 4,7-dihydro-2- [4- (2-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-) hydrochloride methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine The title compound was obtained in the same manner as in Example 1. Mp 213-215 ° C.
Example 12 Example 12-1 Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- [(1-hydroxycyclopropyl) ) carbonylamino] phenyl] -4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (0.57 g, 1.0 mol) in dichloromethane (10 mL) were added diisopropylethylamine (0.52 g, 4 mmol) and 2-hydroxycyclopropanecarboxylic acid (0.204 g, 2 mmol), followed by stirring under the conditions of cooling with ice. To this solution was added benzotriazol-1-iloxitrisdimethylaminophosphonium hexafluorophosphate (BOP reagent) (1.76 g, 4 mmol). After stirring under conditions of ice cooling for 1 hour, the solution was subsequently stirred at room temperature for 4 days. The reaction mixture was concentrated to dryness under reduced pressure, and then the residue obtained was partitioned between water (50 mL) and chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO4) the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel and recrystallized from ether, to give yellow powdered crystals 80.27 g, 41%). XH-NMR (300MHz, CDC13) d: 1. l € -l.20 (2H, m), 1.18 (6H, d), 1.48-1.51 (2H, m), 2.09 (3H, s), 3.64 (2H , s), 3.95 (lH, br s), 4.14. { 2H, s), 4.12-4.19 (lH, m), 5.20 (2H, s), 6.99 (2H, t), 7.10-7.25 (5H, m), 7.34-7.46 (lH, m), 7.57 (2H, d), 7.70 (2H, d), 8.21 (1H, s), 8.82 (1H, s).
Example 12-2: Production of 3- hydrochloride. { N-benzyl-N-ethylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothiene [2 , 3-b] pyridine The title compound, white powdered crystals 80.12 g) were obtained in the same manner, as in Example 1. Solvent for recrystallization: ethanol: ether mp 220-222 ° C. Elemental analysis for C37H35N304SF2 HCl 0.5H20 C (%) H (%) N (%): Calculated: 63.37; 5.32; 5.99 Found: 63.35; 5.24; 5.82 Example 13 Example 13-1: Production of 47-dihydro-2- [4- (3-hydroxy-3-methylbutyrylamino) phenyl] -7- (2,6-ii ".uorcfca? Cyl) -3- (N- benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (1.14 g, 2.0 mmol) in dichloromethane (10 mL) were added diisopropylethylamine (1.04 g, 8 mmol) and 3-hydroxy-3-methylbutanoic acid (0.47 g, 4 mmol). ), followed by stirring under the conditions of ice cooling. To this solution was added benzotriazol-1-yloxytrimethylaminophosphonium hexafluorophosphate (BOP reagent) (1.76 g, 4 mmol). After stirring under the conditions of ice cooling for 1 hour, the solution was subsequently stirred at room temperature for 4 days. The reaction mixture was concentrated to dryness under reduced pressure, and then, the obtained residue was partitioned between water (50 mL) and chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO 4), the solvent was distilled under reduced pressure. The obtained residue was subjected to chromatography on silica gel and recrystallized from ether, to give powdered yellow crystals (0.50 g, 37%). 1H-NMR (300MHz, CDC13) d: 1.17 (6H, d), 1.39 (6H, s), 2.11 (3H, s), 2.58 (2H, s), 3.12 (1H, br s), 3.65 (2H, s), 4.12-4.19 (lH, m), 4.18 (2H, s), 5.27 (2H, s), 7.00 (2H, t), 7.10-7.23 (5H, m), 7.32-7.44 (lH, m) , 7.61 (2H, d), 7.79 (2H, d), 8.24 (1H, s), 8.28 (1H, s).
Example 13-2: Production of 4,7-dihydro-2- [4- (3-hydroxy-3-methylbutyryl-1-amino) phenyl] -7- (2,6-difluorobenzyl) -3- hydrochloride (N -benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine The title compound, powdered white crystals 80.42 g) were obtained in the same manner as in Example 1. Solvent for recrystallization: ethanol mp 216-218 ° C. Elemental analysis for C38H39N3? 4SF2 HCl C H20. { %) H. { %) N (%) Calculated: 62.84: 5.83; 5.79 Found: 62.70; 5.75; 5.82 Example 14 The following compounds (Examples 14-1 to 14-10) were produced by the same method as described in Examples 9, 11, 12 and 13, using the compound obtained in Reference Example 29, and also using several carboxylic acid compounds instead of, for example (2R) -t-butoxy-2-methylpropanoic acid, which was used in Example 9.
Example 14-1: Yield: 46% Amorphous 1H-NMR (300MHz, CDC13) d: 1.18 (6H, d), 1.25 (3H, d), 1.30 (9H, s), 2. 11 (3H, s), 2.60 (lH, br), 3.51 (1H, similar at), 3.59-3.65 (lH, m), 3.65 (2H, s), 4.17 (lH, q), 4.18 (2H, s ), 5.27 (2H, s), 7. 00 { 2H, t), 7.12-7.22 (5H, m), 7.38-7.44 (lH, m), 7.60. { 2H, d), 7.75 (2H, d), 8.28 (1H, s), 9.00 (1H, s).
Example 14-2; Performance: 47% Mp 145-150 ° C.
Example 14-3: Yield: 99% Mw 96-38 ° C.
Example 14-4: Yield 57%: Mw 92-94 ° C.
Example 14-5: Rendimi ent o: 71 'Pf 57-62 ° C Example 14-6 Rendimi ent o: 92% Mw 79-81 ° C.
Example 14-7: Yield: 97% Mp 91-96 ° c.
Example 14-8: Yield: 91% Mw 75-80 ° C.
Example 14-9: Rendimi ent o: 87% Pf 83-88 ° C.
Example 14-10: Yield: 50% Mw 130-135 ° C.
Example 15 Using the compound obtained in Example 14, the following compounds were obtained (Examples 15-1 to 15-6), in the same manner as in Example 10.
Example 15-1: Yield: 69% Mp 161-162 ° C.
Example 15-2 Yield: 94% Mw 153-155 ° C.
Example 15-3: Yield: 62% Mp 224-226 ° C.
Example 15-4: Yield: 56% Mw 230-232 ° C.
Example 15-5: Rendimi at t o: 90% Mw 92-94 ° C.
Example 15-6: Yield: 34% Mw 107-112 ° C.
Example 16 Example 16-1: Production of 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-N'-ethoxyureidophenyl) 4-Oxothieno [2,3-b] pyridine To a solution of the compound obtained in Reference Example 29 (0.40 g, 0.7 mmol) in dichloromethane (15 mL) s added, triethylamine (0.20 mL, 1.43 mmol) and N, N'-carbonyldiimidazole (0.228 g, 1.41 mmol), followed by stirring under the conditions of an ice-cooling, then, the solution was allowed to warm to room temperature and then further stirred for 2.5 days, to the mixture O-methylhydroxylamine hydrochloride (0.59 g, 7.06 mmol) and triethylamine (0.98 mL, 7.03 mmol) were added with cooling with ice, while allowing the solution to return to room temperature, stirring for 1.5 hours. After dilution with water (50 mL), the reaction mixture was extracted with chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel, to give amorphous yellow powders. . { 0.349 g, 77%).
^ -NMR (300MHz, CDC13) d: 1.19 (6H, d), 2.17 (3H, s), 3.72 (2H, brs), 3.83 (3H, s), 4.10-4.20 (lH, m), 4.22 (2H , brs), 5.28 (2H, s), 7.00 (2H, t), 7.15-7.24 (5H, m), 7.37-7.46 < 2H, m), 7.59 (2H, d) I 7.75 (2H, d), 7.83 (1H, br), 8.31 (lH, s).
Example 16-2: Production of 4,7-dihydro-2- [4-N'-methoxyureidophenyl) -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl hydrochloride -4-oxothieno [2, 3-b] pyridine The title compound, white amorphous powders (0.335 g), was obtained in the same manner as in Example 1. Solvent for recrystallization: ethyl acetate-ether Pf 225-230 ° C. mp 225-230 ° C. Elemental analysis for C35H34N40SF2 HCl 0.5H20 C (%) H (%) N (%) Calculated: 60.91; 5.26; 8.12 Found: 60.80; 5.07: 8.17 Example 17 Example 17-1: Production of (R) -4,7-dihydro-2- [4- (2, 3-dihydroxypropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl) -N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine (R) - (+) -2,2-Dimethyl-1,3-dioxolan-4-carboxylic acid methyl ester (1.00 g, 6.24 mmol) was added to a 2N aqueous solution of potassium hydroxide, followed by low stirring. the conditions of a cooling with ice for 3 hours. After neutralization with hydrochloric acid, the solution was precipitated and extracted with ethyl acetate (50 mL) to give, acid (R) - (+) -2, 2-dimethyl-1-1, 3-dioxolan-4 -carboxylic as colorless amorphous powders (0.72 g). To a solution of the compound obtained in Reference Example 29 (0.572 g, 1.0 mmol) in dichloromethane (5 mL) were added diisopropylethylamine (0.04 g, 8 mmol) and the (R) - (+) - 2, 2-dimethyl-l, 3-dioxolan-4-carboxylic acid obtained (0.292 g) , 2 mmol), followed by stirring under the conditions of ice cooling. to this solution was added benzotriazol-1-yloxytrimethylaminophosphonium hexafluorophosphate (BOP reagent) (0.885 g, 2 mmol). After stirring under the conditions of ice cooling for 1 hour, the solution was further stirred at room temperature for 4 days. The reaction mixture was concentrated to dryness under reduced pressure, and then, the obtained compound was partitioned between water (50 mL) and chloroform (50 mL). The aqueous layer was extracted again with chloroform (10 mL). The combined extracts were washed with saline, and dried (MgSO 4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give colorless amorphous powders (0.5 g), which were then dissolved in trifluoroacetic acid (10 mL) and stirred at room temperature for 16 hours. The reaction mixture was concentrated to dryness under reduced pressure and treated with an alkali, after which it was partitioned between chloroform (50 mL) and water (50 mL), and then, the aqueous layer was extracted again with chloroform ( 10 mL). The combined extracts were washed with saline, and dried (MgSO4), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give 0.5 g of amorphous colorless powder (76%). • -H-NMR (300MHz, DMSO-de) d: 1.09 (6H, d), 1.95. { 3H, s), 3.51 (2H, br), 3.55-3.72 (2H, m), 4.01-4.08 (4H, m), 4.84 (lH, t), 5.63 (2H, brs), 5.80 (lH, d) , 7.15-7.24 (7H, m), 7.49-7.57 (lH, m), 7.66 (2H, d), 7.84 (2H, d), 8.53 (lH, s), 9.82 (lH, s).
Example 17-2: Production of (R) -4,7-dihydro-2- [4- (2, 3-dihydroxypropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl) hydrochloride -N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2, 3-b] pyridine The title compound, white powdered crystals (0.43 g) were obtained in the same manner as in Example 1. Solvent for recrystallization: ether-ethanol Mp 146-148 ° C.
Example 18 Production of 4,7-dihydro-2- (4-N'-hydroxy-N 'methylureidophenyl) -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl -4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 29 (1.0 g, 1.75 mmol) in dichloromethane. { 40 mL) were added triethanolamine (0.49 mL, 3.50 mmol) N, N'-carbonyldiimidazole (0.567 mg, 3.50 mmol), followed by stirring under the conditions of ice-cooling, then, the solution was allowed to warm to the At room temperature and subsequently stirred for 17 hours, N-methylhydroxylammonium chloride (730 mg, 8.75 mmol) and triethylamine (1.23 mL, 8.75 mmol) were added to the mixture with ice cooling, while the solution was allowed to warm to room temperature. the room temperature was stirred for 5 hours. The reaction mixture was partitioned between chloroform (300 mL) and aqueous sodium bicarbonate (saturated, 200 mL), and then, the organic layer was washed again with saline (200 mL). After the organic layer was dried (MgSO), the solvent was distilled under reduced pressure. The residue obtained was subjected to chromatography on silica gel to give yellow amorphous powder (0.38 g, 34%). - •• H-NMR (300MHz, CDC13) d: 1.18 (6H, d), 2.15 (3H, s), 3.26 (3H, s), 3.71. { 2H, s), 4.19 (2H, s), 5.25 (2H, s), 7.00. { 2H, t), 7.22- 7.26 (5H, m), 7.41 (1H, m), 7.49 (2H, d), 7.60 (2H, d), 8.06 ( 1H, s), 8.26 (1H, s). Elemental analysis for C35H34N404SF2 0. 8H2O C (%) H (%) N (%) Calculated: 63.78; 5.44; 8.50 Found: 63.73; 5.24; 8.41 Example 19 Production of 2- [4 - [(l-acetoxycyclopropyl) carbonylamino] phenyl] -3- (N-benzyl-N-methylaminophenyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4, 7- dihydro-4-oxothieno [2, 3-b] pyridine To a solution of the compound obtained in Reference Example 28 (0.47 g, 0.7 mmol) and triethylamine (0.08 g, 0.8 mmol) in dichloromethane (10 mL), was added 1-acetoxycyclopropanecarbonyl chloride (0.13 g, 0.8 mmol) with Upon ice cooling, the mixture was stirred at 0 ° C for 30 minutes and at room temperature for 30 minutes. The reaction mixture was diluted with chloroform (20 mL) and the solution was washed with aqueous sodium bicarbonate (saturated, 20 mL), and brine. { 20 mL). After the organic layer was dried (MgSO 4), the solvent was distilled under reduced pressure. The residue was chromatographed on silica gel to give yellow amorphous powders (0.34 g, 66%). 1H-NMR (300MHz, CDC13) d: 1.22 (2H, dd), 1.68 (2H, dd), 2.12 (3H, s), 2.21 (3H, s), 3.64 (2H, s), 4.13 (2H, s) ), 5.28 (2H, s), 7.01 (2H, t), 7.17-7.23 (5H, m), 7.37-7.44 (3H, m), 7.51-7.56 (1H, m), 7.60 (2H, d), 7.81 (2H, d), 7.87-7.90 (3H, m), 7.94 (1H, s). FAB-Mass m / z 732 (MH) + Example 20 Using the compound obtained in the Example of Reference 28, the following compound was obtained, Example 20-1 in the same manner as in Example 19 Example 20-1: Yield: 80% 1H-NMR (300MHz, CDC13) d: 1.77 (6H, s), 2.12 (3H, s), 2.17 (3H, s), 3.63 (2H, s), 4.13. { 2H, s), 5.29. { 2H, s), 7.01 (2H, t), 7.15-7.23 (5H, m), 7.38-7.45 (3H, m), 7.51-7.56 (1H, m), 7.64. { 2H, d), 7.85 (2H, d), 7.88-7.90 (3H, m), 7.94 (1H, s). FAB-Mass m / z 734 (MH) + Using the compound obtained in Reference Example 28, the following compound was obtained, Example 20-2 in the same manner as in Example 19 Example 20-2; Yield: 84% -? - NMR (300MHz, CDC13) d: 1.25 (3H, d), 1.30 (9H, s), 2.10 (3H, s), 2.65-2.67 (lH, br m), 3.51 (2H , t), 3.60 (2H, s), 4.12 (2H, s), 5.28 (2H, s), 7.00 (2H, t), 7.16-7.23 (5H, m), 7.37-7.42 (3H, m), 7.50-7.53 (1H, m), 7.61 (2H, d), 7.80 (2H, d), 7.89 (2H, d), 7.9 (1H, s), 9 .01 (1H, s). FAB-Mass m / z 748 (MH) + Example 21 Production of 3- (N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-2- [4- [(1-hydroxycyclopropyl) carbonyl] phenyl ] -4-oxothieno [2, 3-b] pyridine A mixture of the compound obtained in Example 19 (0.24g, 0.33 mmol) and sodium hydroxide (0.07 mL) in ethanol (8 mL) was stirred at room temperature for 1 hour. Then, the solvent was distilled under reduced pressure, the residue obtained was dissolved in chloroform (30 mL) and the solution was washed with aqueous sodium bicarbonate (Saturated, L) and brine (20 mL). After, the organic layer was dried (MgSO), the solvent was distilled under reduced pressure and the residue was recrystallized from chloroform-ether, to give pale yellow powders (0.17 g, 75%). mp 186-188 ° C. XH-NMR (300MHz, CDC13) d: 1.15 (2H, dd), 1.44 (2H, dd), 2.05 (3H, s), 3.56 (2H, s), 4.01 (2H, s), 4.56. { 1H, br s), 5.24 (2H, s), 7.00 (2H, t), 7.14-7.22 (5H, m), 7.41 (3H, t), 7.55 (2H, d), 7.55 (1H, s), 7.70 (2H, d), 7.88 (2H, d), 7.90 (1H, s), 8.88 (1H, s). Elemental analysis for C4OH33N304SF2 1.0H2O C (%) H (%) N (%) Calculated: 67.88; 4.98; 5.34 Found 67.75; 4.70; 5.90 FAB-Mass m / z 690 (MH) + Example 22 Production of 3- (N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-2- [4-. { 2-hydroxy-2-methylpropionylamino) phenyl] -4-oxothieno [2, 3-b] pyridine Using the compound obtained in Example 20-1, the title compound was obtained, in the same manner as in Example 21. Solvent for recrystallization: chloroform-ether mp 222-223 ° C. 1H-NMR (300MHz, CDC13) d: 1.57 (6H, s), 2.09 (3H, s), 2.50 (1H, s>, 3.59 {2H, s), 4.10 (2H, s), 5.28. { 2H, s), 7.01 (2H, t), 7.16-7.25. { 5H, m), 7.38-7.44 (3H,), 7.51-7.56 (1H, m), 7.67 (2H, d), 7.82 (2H, d), 7.89 (2H, d), 7.93 (1H, s), 8.82 (1H, s). FAB-Mass m / z 692 (MH) + Example 23 Production of (R) -3- (N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7-dihydro-2- [4- [. { 3-hydroxy-2-methylbutyrylamino) phenyl] -4-oxothieno [2, 3-b] pyridine Using the compound obtained in Example 20-2, the title compound was obtained in the same manner as in Example 10. Solvent for recrystallization: chloroform-ether mp 145-146 ° C. XH-NMR (300MHz, CDC13) d: 1.25 (3H, d), 2.07 (3H, s), 2.63-2.70 (1H, m), 2.84 (1H, br s), 3.57. { 2H, s), 3.78-3.81 (2H, m), 4.08 (2H, s), 5.27 (2H, s), 7.00 (2H, t), 7.17-7.20 (5H, m), 7.37-7.43. { 3H, m), 7.51-7.56 (1H, m), 7.61 (2H, d), 7.77 (2H, d), 7.89 (2H, d), 7.94 (1H, s), 8.31 (1H, br s).
FAB-Mass m / z 692 (MH) + Preparation Example 1 Using 100 mg of the compound produced in Example 1-2, 165 mg of lactose, 25 mg of corn starch, 4 mg of polyvinyl alcohol and 1 mg of magnesium stearate, tablets are produced by a conventional method.
Preparation Example 2 The compound produced in Example 1-2 (5 g) is dissolved in distilled water for injection, to make a total volume of 100 mL. This solution is aseptically filtered through a 0.22 μm filtration membrane (produced by Sumitomo Electric Industries, Ltd. or Sartorius) and distributed to 2 mL sterile washed ampule, followed by freeze dehydrate by a conventional method, to give a injectable preparation of 100 mg / vial dehydrate by freezing.
Preparation Example 3 Using 100 mg of the compound produced in Example 3, 165 mg of lactose, 25 mg of corn starch, 4 mg of polyvinyl alcohol and 1 mg of magnesium stearate, tablets are produced by a conventional method.
Preparation Example 4 The compound produced in Example 3 (5 g), is dissolved in distilled water for injection, to make a total volume of 100 mL. This solution is aseptically filtered through a 0.22 μm filtration membrane (produced by Sumitomo Electric Industries, Ltd. or Sartorius) and distributed to 2 mL per sterile ampule, followed by dehydration by freezing by a conventional method, to give an injectable preparation of 100 mg / vial dehydrated by freezing.
Preparation Example 5 (1) the compound produced in Example 1-2 5 g (2) Lactose / crystalline cellulose (particles) 330 g (3) D-mannitol 29 g (4) 20 G low substitution hydroxypropyl cellulose (5) ) Talc 25 g (6) Hydroxypropyl cellulose 50 g (7) Aspartame 3 g (8) Dipotassium glycyrrhizinate 3 g. (9) Hidoxypropylmethyl cellulose 2910 30 g (10) Titanium oxide 3.5 g (11) Fiery yellow sesquioxide 0.5 g (12) silicic light anhydride 1 g components (1), (3), (4), (5) , (6), (7) and (8) are suspended or dissolved in purified water and coat the nuclear particles (2), to give a fine base fabric, which is then coated, subsequently with, the components (9). ) to (11) to give coated fabrics, which are then mixed with the composite (12) to give 500 g of 1% of composite tissues. These tissues are divided to 500 mg folded tissues.
Experimental Example 1 Preparation of l25pI-leuprorelin To a tube containing 10 μl of aqueous solution 3xlO-M of leuprorelin and 10 μl of 0.01 mg / mL of lactoperoxidase, 10 μl (37 MBq) of a solution of Na125I was added. After stirring, 10 μl of H202 was added at 0.001%, the reaction was carried out at room temperature for 20 minutes. The reaction was stopped by the addition of 700 μl of a 0.05% solution of TFA (trifluoroacetic acid), followed by purification by reverse phase HPLC. The HPLC conditions used are shown below. I-leuprorelin was eluted at the retention time of 26 to 27 minutes.
Column: TSKgel ODS-80 ™ (TM indicates a registered trademark, same applies below) CTR (4.6 nm x 10 machine controller) Eluents: Solvent A (0.05% TFA) Solvent B (CH3CN 'at 40% -TFA 0.05%) 0 minutes (100% solvent A) - 3 minutes (100% solvent A) - 7 minutes (50% solvent A + 50% solvent B) - 40 minutes. { 100% solvent B) Evasion temperature: Ambient temperature Speed of elusion: 1 mL / min Experimental Example 2 Preparation of a rat pituitary anterior lobe membrane fraction containing receptors GnRH The anterior lobes of the pituitary glands were isolated from forty Wistar rats (8 weeks of age, males), and washed with homogenized buffer [25 mM Tris (tris [hydroxymethyl) aminomethane) -HCl, sucrose 0.3M, EGTA lmM ( glycol ether-ether diamine- N, N, N ', N' -tetracetic acid), PMSF 0.25 mM. { phenylmethylsulfonyl fluoride), 10 U / mL of aprotinin, 1 mg / mL of pepestatin, 20 mg / mL of leupéptin, 100 mg / mL of phosphoramidon, 0.03% sodium azide, pH 7.5]. the pituitary tissue was floated in 2 mL of the buffered-homogenate and homogenized using a Polytron homogenizer. The homogenadp was centrifuged at 700 x g for 15 minutes. The supernatant was recovered in an ultra centrifuge tube and centrifuged at 100,000 x g for 1 hour to provide a pellet of the membrane fraction. This pellet was suspended in 2 mL of the assay buffer [25 mM tris-HCl, 1 mM EDTA. { ethylenediaminetetracetic acid), 1% BSA (bovine serum albumin), 0.25 mM PMSF, 1 mg / mL pepestatin, 20 mg / mL leupeptin, 100 mg / mL phosphoramidon, 0.03% sodium azide, pH 7.5 ) and the suspension was centrifuged at 100,000 xg for 1 hour. The membranous fraction recovered as a pellet was suspended in 10 mL of assay buffer, divided into portions, preserved at -80 ° C and thawed when necessary.
Experimental Example 3 Preparation of the membranous fraction of CHO (ovaries from Chinese hamsters) containing the human GnHR receptor CHO cells expressing the GnRH receptor (EP-A-678577) (109 cells) were suspended in phosphate buffered saline supplemented with 5 nM EDTA (ethylenediaminetetracetic acid) (PBS-EDTA) and centrifuged at 100 xg for 5 minutes. To the cell pellets, 10 mL of a cellular homogenous buffer (NaHC03 lOmM, 5 mM EDTA) was added, followed by homogenization using the Polytron homogenizer. After centrifugation at 400 x g for 15 minutes, the supernatant was transferred to an ultra centrifugation tube and centrifuged at 100,100 x g for 1 hour to give a precipitate of the membranous fraction. This precipitate was suspended in 2 mL of a test buffer and centrifuged at 100,000 x g for 1 hour. The membranous fraction recovered as a precipitate was again suspended in 20 mL of the assay buffer, distributed, and stored at -80dC before use was thawed.
EXPERIMENTAL EXAMPLE 4 Determination of the binding inhibition of 125I-leurporelin The membranous fractions of human and rat prepared in Experimental Examples 2 and 3 were diluted with the assay buffer to give a dilution of 200 mg / mL, the which was distributed to 188 mL per tube. Where the membranous fraction of the rat pituitary anterior lobe was used, 2 ml of a 0.1 mM solution of the compound in 60% DMSO (dimethyl sulfoxide) and 10 μl of 38 nM 125I-leuprorelin were added to each tube simultaneously. Where the cell membrane fraction of the CHO was used with the expressed human GnRH receptors, 2 mL of a 2 mM solution of the compound in 60% DSO and 10 μl of 38 nM 125I-leuprorelin were added to each tube simultaneously. To determine the maximum amount of agglutination, a reaction mixture of 2 μl of 60% DMSO and 10 μl of 38 nM 125I-leuprorelin was prepared. To determine the amount of non-specific binding, a reaction mixture of 2 μl of 100 μM of leuprorelin in 60% solution in DMSO of 38 nM 125I-leuprorelin was prepared.
Where the membranous fraction of the anterior lobe of the bovine or rat pituitary was used, the reaction was conducted at 4 ° C for 90 minutes. Where the membranous fraction of CHO with expressed human GnRH receptors was used, the reaction mixture was carried out at 25 ° C reduced pressure 60 minutes. After each reaction, the reaction mixture was aspirated and filtered through a Whatman glass filter (GF-F) treated with polyethylene diamine. After filtration, the radioactivity of the remaining 125I-leuprpreline was measured on the filter paper, with a counter of y. The expression (TB-SB) / (TB-NSB) xlOO was calculated (where SB = radioactivity with the aggregate compound, TB = determined maximum radioactivity, NSB = determined non-specific radioactivity) to find the inhibition of the binding speed ( %) of each test compound. In addition, the rate of inhibition was determined by varying the concentration of the test substance and 50% of the inhibitory concentration (IC50 value) of the compound was calculated from the Hill plot. The results are shown below.
Inhibition of agglutination value IC 50 Speed test compound (%) (μM) Rat (lμM) Human (20 μM) Human Rat Comp. of Ex .1-2 96 102 0.06 0.0001 Comp. of Ex. 3 | 62 NT 0.6 0.0002 NT: not measured EXPERIMENTAL EXAMPLE 5 Suppression of LH from blood in castrated monkeys The compound produced in Example 1-2 was orally administered to castrated male cynomolgus monkeys (Macaca fascicularis), and the LH of the blood was quantified. The male cynomolgus monkeys, used had 3 years 8 months, 7 years 7 months of age at the time of experimentation, have been neutered for more than half a year prior to the examination. To the test animals [n = 2; compound (1) and compound [2) 1, they were given 30 mg / kg (3 mL / kg) of the compound suspended in 0.5% methyl cellulose at a final concentration of 1%, by oral administration, and the control animals (n = 3, control (1), control (2) and control (3)] were given 3 mL / kg of 5% methyl cellulose dispersant only, by oral administration, at 24 hours and immediately before administration. administration and at 2, 4, 6, 8, 24, and 48 hours after the administration, blood was collected for heparinized plasma samples, via the femoral vein and immediately stored under freezing conditions.
Plasma LH concentrations were determined by a bioassay using mouse testicular cells. Testicular cells were collected from male BALB / C mice (8 to 9 weeks of age) and washed three times with 1 mL of modified Dulbecco's Eagle's medium (DMEM-H) containing 20 mM HEPES and 0.2% BSA. testis After incubation at 37 ° C for 1 hour, the cells were passed through a nylon mesh filter (70 μm) and distributed to test tubes at 8 x 10 5 cells / tube. After, that the cells were washed twice with 0.4 mL of DMEM-H, 0.4 mL of a solution of DMEM-H containing, any Equine LH (Sigma Company) was added, as the standard LH, or monkey plasma, diluted previously up to 1,000 folds, as the test sample, followed by the reaction at 37 ° C for 2 hours. The concentration of testosterone in the culture supernatant was determined by a radioinone test (CIS Diagnostics Company), and the LH concentrations in the plasma of the test monkey were calculated from the standard curve for the equine standard LH.
The results are given together in Figure 1. The indicated compound is the compound obtained in Example 2-1. For control (1), control (2), and control (3), changes over time in the LH concentrations in each animal were expressed as percentage ratios to the respective reference values, which comprise the LH concentrations immediately before administration in each control animal (Cynomolgus monkeys). Similarly, for compound (1) and compound (2), changes over time in the concentrations of LH in each animal (cynomolgus monkeys) that received the compounds of Example 1-2 are expressed as ratios of% to respective reference values, which comprise the LH concentrations immediately before the administration of the compound. The administration time was defined as zero (0), and the hours before and after administration are shown by minus and plus signs respectively.
INDUSTRIAL APPLICABILITY The compound of the present invention possesses excellent gonadotropin releasing hormone antagonizing activity. It is also good in its ability to absorb itself and pharmacokinetics. With low toxicity, it is also excellent in safety.
The compound of the present invention can therefore be used as a prophylactic or therapeutic agent for hormone-dependent diseases etc. Specifically, it is effective as a prophylactic or therapeutic agent for sex hormone-dependent cancers (e.g., prostate cancer,, uterine cancer, breast cancer, pituitary tumor), prostatic hypertrophy, hysteromyoma, endometriosis, precocious puberty, amenorrhea syndrome, syndrome of the multiocular ovary, pimples etc., or as a regulator of pregnancy (for example, contraceptive), remedy against infertility or regulator of menstruation. It is also effective as an animal estrus regulator, meat quality enhancer for feed or animal growth regulator in the field of animal husbandry, and as a promoter of roe deposition in the field of the fishing industry.
It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.
Having described the invention as above, property is claimed as contained in the following:

Claims (24)

REIVTNIDICACINES
1. A compound of the formula: characterized in that, R 1 represents a C 1-7 alkyl group, which may be substituted, a C 3-7 cycloalkyl group, which may be substituted, an alkoxyamino group of C1-6, which may be substituted or a hydroxyamino group, which may be substituted; and R2 represents an alkyl group of C7-7, which may be substituted or a phenyl group, which may be substituted; when R1 is an unsubstituted C1-7 alkyl group, then R 2 is a substituted C 1-7 alkyl group or a substituted phenyl group, or a salt thereof.
2. A compound of claim 1 or a salt thereof, characterized in that, R1 is (1) an alkyl group, which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) acyloxy of C1-7, (iii) amino, which may be substituted by 1 to 2 substituents selected from the group consisting of C6-6-carbonyl alkoxy, benzyloxycarbonyl, Ci-3 acyl, C1-3 alkylsulfinyl, and C-alkyl ? -3, (iv) Ci-3 alkoxy, which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkoxy-carbonyl and C1-3-alkoxy and (v) C-alkoxy? _6-carbonyl, (2) a cycloalkyl group of C3-7 / which may be substituted by 1 to 3 substituents selected from the group consisting of (i) hydroxy, (ii) C1-7 acyloxy, (iii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C?-6-carbonyl alkoxy, benzyloxycarbonyl, C?-3 acyl, C 1-3 alkylsulfonyl and C 1-3 alkyl, (iv) C 1-10 alkoxy , which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkyloxy-carbonyl and C1-3-alkoxy and (v) C6-6-carbonyl-alkoxy, (3) an alkoxyamino group of C? -β, which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) C 1-7 acyloxy, (iii) amino, which may be to be substituted by 1 or 2 substituents selected from the group consisting of C?-6-carbonyl alkoxy, benzyloxycarbonyl, C 1-3 acyl, C 1-3 alkylsulfonyl and C? _3 alkyl, (iv) C 1-10 alkoxy , which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkyloxycarbonyl and C1-3alkoxy and (v) C6-6-carbonyl alkoxy, or (4) a hydroxyamino group, which may be substituted by 1 or 2 substituents selected from the group consisting of (i) C1-7 acyloxy, (ii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C-alkoxy? _6-carbonyl, benzyloxycarbonyl, acyl of C? _, Alkylsulfonyl of C1-3 and alkyl of C1-3, (iii) C1-10 alkoxy, which may be substituted by 1 to 3 substituents selected from the group consisting of C3-7-cycloalkyloxycarbonyl and C1-3alkoxy and (iv) C6-6alkyl; and R 2 is (1) a C 1-7 alkyl group which may be substituted by 1 to 5 substituents selected from the group consisting of (i) hydroxy, (ii) acyloxy, (iii) amino, which may be substituted by 1 or 2 substituents selected from the group consisting of C 1-6 alkoxycarbonyl, benzyloxycarbonyl, C 1-3 acyl, C 1-3 alkylsulfonyl and C 3-3 alkyl, (iv) C 1-10 alkoxy which may be to be substituted by 1 to 3 substituents selected from the group consisting of C3-7-carbonyl cycloalkyloxy and C1-3 alkoxy and (v) C6-6-carbonyl alkoxy, or (2) a phenyl group, which may be to be substituted by 1 to 5 substituents selected from the group consisting of halogen, C? _3 alkyl and C1-3 alkoxy.
3. A compound of claim 1 or a salt thereof, characterized in that R1 is an alkyl group of Ci-7, which may be substituted or a cycloalkyl group of C3-7 / which may be substituted.
4. A compound of claim 1 or a salt thereof, characterized in that, R1 is a substituted, branched C3_7 alkyl group, or a substituted C3_7 cycloalkyl group.
5. A compound of claim 1 or a salt thereof, characterized in that, R1 is an alkyl group of Ci-7, substituted by hydroxy, or a cycloalkyl group of C3-7 substituted by hydroxy.
6. A compound of claim 1 or a salt thereof, wherein R 1 is a substituted C 3-7 cycloalkyl group.
7. A compound of claim 1 or a salt thereof, characterized in that, R 1 is a cyclopropyl group, which may be substituted by hydroxy.
8. A compound of claim 1 or a salt thereof, characterized in that R2 is a branched C3-7 alkyl group, which may be substituted.
9. A compound of claim 1 or a salt thereof, characterized in that, R 2 is a phenyl group, which may be substituted.
10. A compound of claim 1 or a salt thereof, characterized in that R2 is a phenyl group.
11. A compound of claim 1 or a salt thereof, characterized in that, R1 is a cycloalkyl group of C3-7 and R2 is an alkyl group of Ci-β.
12. A compound of claim 1 or a salt thereof, characterized in that, R1 is (l) a C1-4 alkyl group, substituted by 1 or 2 hydroxy, (2) a C3-7 cycloalkyl group substituted by hydroxy, or (3) a C1-3 alkoxyamino group; and R2 is an isopropyl group or a phenyl group,
13. A compound of claim 1 or a salt thereof, characterized in that, R1 is (1) an alkyl group of C? -7, which may be substituted by 1 or 2 substituents de-selected from the group consisting of hydroxy, C? -3-carbonyloxy, amino, benzyloxycarbonylamino, C? _3 alkoxy, C? -3-C3-alkoxy alkoxy and C? _3-carbonyl alkoxy, (2) a C3_7 cycloalkyl group, which can to be substituted by a hydroxy, or a C?-3-varbonyloxy alkyl, or (3) an alkoxyamino group of Ci--3; and R2 is (1) an isopropyl group, which may be substituted by a hydroxy or (2) a phenyl group.
14. 3- (N-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4-oxothiene [2, 3-b] pyridine or a salt thereof.
15. 3- (N-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -4 -oxotiene [2, 3-b] pyridine or a salt thereof.
16. 3- (N-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (-2,6-difluorobenzyl) -2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4 -oxotiene [2, 3-b] pyridine or a salt thereof.
17. 3- (N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl-4,7-dihydro-2- [4- [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothiene [2, 3-b] pyridine or a salt thereof.
18. A process for the production of a compound of claim 1 or a salt thereof, characterized in that it comprises the reaction of a compound of the formula: characterized in that R2 represents a C1-7 alkyl group, which may be substituted or a phenyl group, which may be substituted, or a salt thereof, with i) a compound of the formula: RlaCOOH wherein Rla represents a C 1-7 alkyl group, which may be substituted or a C 1-3 cycloalkyl group, which may be substituted, or a salt thereof or a reagent derived therefrom; or ii) carbonyldiimidazole, phosgene or a chloroformate, followed by the reaction with a compound of the formula: RlbH wherein Rlb represents an alkoxyamino group of C? -β, which may be substituted or a hydroxyamino group, which may be substituted , or a salt thereof.
19. A process for producing a compound of claim 3 or a salt thereof, characterized in that it comprises the reaction of a compound of the formula: wherein R represents an alkyl group of C? _7, which may be substituted or a phenyl group, which may be substituted, or a salt thereof, with a compound of the formula: RlaCOOH wherein Rla represents a C 1-7 alkyl group, which may be substituted or a C 3-7 cycloalkyl group, which may be substituted, or a salt thereof or a reagent derived therefrom.
20. A pharmaceutical composition, characterized in that it comprises a compound of claim 1 or a salt thereof.
21. A pharmaceutical composition of claim 20, characterized in that it is for antagonizing the gonadotropin releasing hormone.
22. A pharmaceutical composition of claim 21, characterized in that it is for preventing or treating a sex hormone dependent disease.
23. A method for antagonizing gonadotropin releasing hormone, in a mammal in need thereof, characterized in that it comprises administering to said mammal an effective amount of a compound of claim 1 or a salt thereof with a pharmaceutically acceptable excipient. , carrier or diluent.
24. The use of a compound of claim 1 or a salt thereof to manufacture a pharmaceutical composition for antagonizing the gonadotropin releasing hormone.
MXPA/A/2000/011184A 1998-06-26 2000-11-14 Thienopyridine compounds, their production and use MXPA00011184A (en)

Applications Claiming Priority (2)

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JP10/181263 1998-06-26
JP10/333004 1998-11-24

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MXPA00011184A true MXPA00011184A (en) 2001-09-07

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