USRE49111E1 - 2-acylaminothiazole derivative or salt thereof - Google Patents

2-acylaminothiazole derivative or salt thereof Download PDF

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USRE49111E1
USRE49111E1 US16/856,738 US201516856738A USRE49111E US RE49111 E1 USRE49111 E1 US RE49111E1 US 201516856738 A US201516856738 A US 201516856738A US RE49111 E USRE49111 E US RE49111E
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alkyl
compound
methyl
thiazol
bladder
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US16/856,738
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Taisuke Takahashi
Takanori Koike
Kenji Negoro
Hiroaki Tanaka
Jun Maeda
Kazuhiro Yokoyama
Hajime Takamatsu
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Asahi Pharma Co Ltd
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Astellas Pharma Inc
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to a 2-acylaminothiazole derivative or a salt thereof which is useful as an active ingredient for a pharmaceutical composition, in particular, a pharmaceutical composition for treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M 3 receptor.
  • the important roles of the lower urinary tract are urine storage and voiding, which are regulated by a coordinated action of the bladder and the urethra. That is, during urine storage, the bladder smooth muscle is relaxed and the urethral sphincter is contracted, whereby a state in which urethral resistance is high is maintained and urinary continence is maintained. On the other hand, during voiding, the bladder smooth muscle is contracted, the urethra smooth muscle is relaxed, and contraction of the external urethral sphincter is also inhibited.
  • Examples of the lower urinary tract disorder include urine storage dysfunction such as overactive bladder, in which urine cannot be retained during urine storage, and voiding dysfunction, in which urine cannot be drained sufficiently during voiding due to an increase in the urethral resistance or a decrease in the bladder contractile force. These two disorders may develop simultaneously in some cases.
  • Voiding dysfunction is caused by a decrease in the bladder contractile force or an increase in urethral resistance during voiding, and causes difficulty in voiding, straining during voiding, a weak urine stream, extension of voiding time, an increase in residual urine, a decrease in voiding efficiency, or the like.
  • the decrease in the bladder contractile force during voiding is referred to as underactive bladder, acontractile bladder, or the like.
  • As a factor causing such a decrease in the bladder contractile force during voiding for example, aging, diabetes mellitus, benign prostatic hyperplasia, neurological diseases such as Parkinson's disease and multiple sclerosis, spinal cord injury, neurological disorders by pelvic surgery, and the like have been known (Reviews in Urology, 15; pp. 11-22 (2013)).
  • the muscarinic receptors are currently classified into five subtypes, M 1 , M 2 , M 3 , M 4 , and M 5 , and it has been known that the subtypes involving the contraction in the bladder smooth muscle is mainly M 3 (Pharmacological Reviews, 50; pp. 279-290 (1998); The Journal of Neuroscience, 22; pp. 10627-10632 (2002)).
  • bethanechol chloride which is a non-selective muscarinic receptor agonist
  • distigmine bromide which is a cholinesterase inhibitor
  • these drugs have cholinergic side effects such as diarrhea, abdominal pain, and perspiration.
  • cholinergic crisis is occurred as a serious side effect, which require attention during use (Uhretid (registered trademark), tablet 5 mg, package insert, Torii Pharmaceutical Co., Ltd., and Besacholine (registered trademark) powder 5%, package insert, Eisai Co., Ltd.).
  • voiding dysfunction associated with benign prostatic hyperplasia has been well-known, which is characterized in that the urethra is partially occluded by nodular enlargement of the prostatic tissue.
  • an adrenergic a, receptor antagonist has been used as a therapeutic drug for voiding dysfunction associated with benign prostatic hyperplasia (Pharmacology, 65; pp. 119-128 (2002)).
  • residual urine after voiding may be observed in some cases.
  • the increased residual urine may cause a decrease in effective bladder capacity, and thus cause overactive bladder symptoms such as urinary frequency or severe symptoms such as hydronephrosis in some cases.
  • Patent Document 1 discloses that a compound represented by the following general formula (A) including a compound of the formula (A1) below, which is disclosed in Example 315, has a Ba/F3 cell proliferative activity through a human c-mycloproliferative leukemia virus type P (c-Mpl), and has thrombocyte increasing activity.
  • Patent Document 2 discloses that a compound represented by the following general formula (B) has an AMPK pathway activating action.
  • Ring B represents a heteroarylene or the like
  • J represents —NR 13 C(O)— or the like
  • D 1 , D 2 and D 3 each represent N, CH, or the like
  • E represents —NR 1 R 2 or the like
  • R 1 and R 2 may be combined with an adjacent nitrogen atom to form a heterocycloalkyl which may be substituted.
  • Non-Patent Document 1 discloses that a compound represented by the following formula (C1) is an allosteric enhancer of a muscarinic M 3 receptor.
  • Non-Patent Document 2 discloses that WIN 62,577 represented by the following formula is a rat NK1 receptor antagonist and, at the same time, an allosteric enhancer of a muscarinic receptor.
  • the present invention provides a novel compound which is expected as an active ingredient for a pharmaceutical composition, in particular, for a pharmaceutical composition for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor, which acts as a muscarinic M 3 receptor-positive allosteric modulator.
  • a thiazole derivative substituted with pyrazinylcarbonylamino at the 2-position is an excellent muscarinic M 3 receptor-positive allosteric modulator and is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor, thereby completing the present invention.
  • the present invention relates to a compound of the formula (I) or a salt thereof, and a pharmaceutical composition comprising a compound of the formula (I) or a salt thereof and an excipient.
  • R 1 is —N(—R 11 )(—R 12 ), or cyclic amino which may be substituted,
  • R 11 is C 1-6 alkyl
  • R 12 is C 1-6 alkyl which may be substituted, or C 3-8 cycloalkyl which may be substituted,
  • R 2 is aryl which may be substituted, monocyclic aromatic hetero ring which may be substituted, or bicyclic aromatic hetero ring which may be substituted,
  • R 3 's are the same as or different from each other, and are each C 1-6 alkyl
  • W is C 1-6 alkylene
  • n is an integer of 0 to 4.
  • Patent Document 1 does not disclose a specific compound which is a compound of the formula (A) wherein R 3 is pyrazinyl, and neither discloses nor suggests an action on a muscarinic receptor or an action on bladder/urethral diseases.
  • Patent Document 2 does not disclose a specific compound which is a compound of the formula (B) wherein ring B is thiazole, and neither discloses nor suggests an action on a muscarinic receptor or an action on bladder/urethral diseases.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the formula (I) or a salt thereof, and a pharmaceutically acceptable excipient.
  • the present invention relates to a pharmaceutical composition for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor, comprising the compound of the formula (I) or a salt thereof.
  • the present invention relates to an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor, comprising the compound of the formula (I) or a salt thereof.
  • the present invention relates to use of the compound of the formula (I) or a salt thereof for the manufacture of a pharmaceutical composition for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M 3 receptor, use of the compound of the formula (I) or a salt thereof for preventing or treating bladder/urinary tract diseases related to bladder contractions via a measuring M 3 receptor, the compound of the formula (I) or a salt thereof for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M 3 receptor, and a method for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M 3 receptor, comprising administering to a subject an effective amount of the compound of the formula (I) or a salt thereof.
  • the “subject” is a human or a non-human animal in need of the prevention or treatment, and in one embodiment, a human in need of the prevention or treatment.
  • the compound of the formula (I) or a salt thereof is a muscarinic M 3 receptor-positive allosteric modulator, and can thus be used as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor.
  • the positive allosteric modulator is a compound which binds to an allosteric site different from a ligand binding site, and has an effect of increasing the affinity of an agonist to a receptor by mainly causing a structural change in a receptor, and thus changing the signal level of agonistic activity.
  • the positive allosteric modulator does not exhibit an agonistic effect by itself, and increases the effect of an endogenous agonist.
  • Examples of the advantages of positive allosteric modulator over the agonists include (1) avoiding the side effects since the positive allosteric modulator exhibits an enhancement in the endogenous agonist stimulation dependently, (2) having a possibility of obtaining high subtype selectively since the positive allosteric modulator binds to a site other than a ligand binding site, and (3) less probability of causing desensitization, which can be seen with the agonists (Pharmacological Reviews, 63; pp. 59-126 (2011)).
  • the muscarinic M 3 receptor-positive allosteric modulator means a compound which enhances an effect via the muscarinic M 3 receptor by an agonist stimulation-dependent or nerve stimulation-dependent manner. Accordingly, only during voiding, the effect on enhancing bladder contraction is expected and the muscarinic M 3 receptor-positive allosteric modulator is possibly useful as an agent for improving various symptoms associated with voiding dysfunction. Further, by such a specific action during voiding, it is expected that it is possible to avoid cholinergic side effects, known to be induced with bethanechol chloride and distigmine bromide.
  • the muscarinic M 3 receptor-positive allosteric modulator increases bladder contractile force during voiding, an effect in voiding dysfunction which is caused by an increase in urethral resistance can also be expected. A decrease in residual urine by such improvement of voiding dysfunction leads to an increase in the effective bladder capacity, and thus, it can be expected to improve urine storage functions as well as to avoid renal disorder.
  • the muscarinic M 3 receptor-positive allosteric modulator is expected to be useful as an agent for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M 3 receptor.
  • the present inventors have newly discovered a compound that acts as the modulator, thereby completing the present invention.
  • examples of the “bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor” include voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrustor underactivity, neurogenic bladder, urethra relaxation failure, detrustor-external urethral sphincter dyssynergia, overactive bladder, urinary frequency, nocturia, urinary incontinence, benign prostatic hyperplasia, interstitial cystitis, chronic prostatitis, urethral calculus, or the like, preferably, voiding dysfunction or urine storage dysfunction in underactivity bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, and neurogenic bladder.
  • the “alkyl” is linear alkyl and branched alkyl. Accordingly, the “C 1-6 alkyl” is linear or branched alkyl having 1 to 6 carbon atoms, and specific examples thereof include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl; in one embodiment, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, each of which is C 1-4 alkyl; in one embodiment, a group selected from the group consisting of methyl, ethyl, isopropyl, and isobutyl; and in one embodiment, a group selected from the group consisting of methyl and ethyl.
  • alkylene is linear alkylene or branched alkylene.
  • C 1-6 alkylene is linear or branched alkylene having 1 to 6 carbon atoms, and examples thereof include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, propylene, methylmethylene, ethylethylene, 1,2-dimethylethylene, or 1,1,2,2-tetramethylethylene; in one embodiment, C 1-3 alkylene; in one embodiment, methylene or ethylene; in one embodiment, methylene; and in another embodiment, ethylene.
  • halogeno-C 1-6 alkyl is C 1-6 alkyl substituted with at least one halogen atom; in one embodiment, C 1-6 alkyl substituted with 1 to 5 halogen atoms; in one embodiment, difluoromethyl or trifluoromethyl; and in one embodiment, trifluoromethyl.
  • the “cycloalkyl” is a saturated hydrocarbon cyclic group.
  • the “C 3-8 cycloalkyl” is a saturated hydrocarbon cyclic group having 3 to 8 ring members, and specific examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl; in one embodiment, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is C 3-6 cycloalkyl; and in one embodiment, cyclopropyl.
  • the “aryl” is a C 6-14 monocyclic to tricyclic aromatic hydrocarbon cyclic group and includes a partially hydrogenated cyclic group thereof, and specific examples thereof include phenyl, naphthyl, tetrahydronaphthyl, indanyl, or indenyl; and in one embodiment, phenyl.
  • the “monocyclic aromatic hetero ring” is a monocyclic aromatic hetero ring group having 5 to 7 ring members, which has 1 to 4 hetero atoms selected from the group consisting of a nitrogen atom, an oxygen atom, and a sulfur atom as a ring-constituting atom, and specific examples thereof include pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furyl, thienyl, oxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, or azepanyl; in one embodiment, thienyl or pyridyl; and in one embodiment, thienyl.
  • the “bicyclic aromatic hetero ring” is a bicyclic aromatic hetero ring group in which the monocyclic aromatic hetero ring is fused with a benzene ring or monocyclic aromatic hetero ring and includes a partially hydrogenated ring group thereof, and specific examples thereof include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzofuranyl, benzothienyl, benzooxazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridyl, thienopyridyl, indolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl, tetrahydroisoquinolyl, dihydrofuropy
  • the “saturated hetero ring” is a 3- to 8-membered saturated ring group, which has 1 to 4 hetero atoms selected from the group consisting of a nitrogen atom, an oxygen atom, and a sulfur atom as a ring-constituting atom, and may be bridged with C 1-6 alkylene, in which a sulfur atom as the ring-constituting atom may be oxidized.
  • azepanyl diazepanyl, oxazepanyl, thiazepanyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, piperidinyl, pyrazolidinyl, piperazinyl, azocanyl, thiomorpholinyl, thiazolindinyl, isothiazolindinyl, oxazolindinyl, morpholinyl, thiomorpholinyl, tetrahydrothiophenyl, oxathioranyl, oxiranyl, oxetanyl, dioxiranyl, tetrahydrofuranyl, tetrahydropyranyl, and 1,4-dioxanyl.
  • the “cyclic amino” is a 4- to 7-membered group having a bond at a ring-constituting nitrogen atom in the saturated hetero ring.
  • Specific examples thereof include aziridin-1-yl, azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, azocan-1-yl, morpholin-4-yl, thiomorpholin-4-yl, piperazin-1-yl, 1,4-diazepan-1-yl, 1,4-oxazepan-4-yl, or 1,4-thiazepan-4-yl; in one embodiment, pyrrolidin-1-yl, piperidin-1-yl, azetidin-1-yl, morpholin-4-yl, or piperazin-1-yl, and in one embodiment, pyrrolidin-1-yl or piperidin-1-yl.
  • halogen means fluoro, chloro, bromo, or iodo; in one embodiment, fluoro, chloro, or bromo; in one embodiment, fluoro or chloro; in one embodiment, fluoro; and in another embodiment, chloro.
  • the expression “which may be substituted” means “which is not substituted” or “which is substituted with 1 to 5 substituents”. Further, if it has a plurality of substituents, the substituents may be the same as or different from each other.
  • Examples of the acceptable substituent in the “cyclic amino which may be substituted”, the “C 3-8 cycloalkyl which may be substituted”, the “aryl which may be substituted”, the “monocyclic aromatic hetero ring which may be substituted”, and the “bicyclic aromatic hetero ring which may be substituted” include substituents in the following Group G.
  • Examples of the substituent in the “cyclic amino which may be substituted” further include oxo ( ⁇ O).
  • Examples of the preferable substituents for the “cyclic amino which may be substituted” in R 1 include, in one embodiment, the substituents described in (a) to (c), (f), and (g) of Group G above; in one embodiment, C 1-6 alkyl which may be substituted with at least one group selected from the group consisting of —OH, —O—(C 1-6 alkyl), —CN, —SO 2 —(C 1-6 alkyl), and halogen; in one embodiment, a group selected from the group consisting of C 1-6 alkyl and halogeno-C 1-6 alkyl; and in one embodiment, a group selected from the group consisting of methyl and ethyl.
  • Examples of the preferable substituents for the “C 1-6 alkyl which may be substituted” in R 12 include, in one embodiment, the substituents described in (b) to (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C 3-8 cycloalkyl, —O—(C 1-6 alkyl), —O—(C 3-8 cycloalkyl), halogen, —CN, and cyclic amino; in one embodiment, a group selected from the group consisting of C 3-8 cycloalkyl and —O—(C 1-6 alkyl); and in one embodiment, a group selected from the group consisting of cyclopropyl and methoxy.
  • Examples of the preferable substituents for the “C 3-8 cycloalkyl which may be substituted” in R 13 include, in one embodiment, the substituents described in (a) to (c), (f), and (g) of Group G above; and in one embodiment, C 1-6 alkyl which may be substituted with —O—(C 1-6 alkyl).
  • Examples of the preferable substituents for the “aryl which may be substituted” in R 2 include, in one embodiment, the substituents described in (a) to (d), (f), (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C 1-6 alkyl, halogen-C 1-6 alkyl, —O—(C 1-6 alkyl), —O-(halogeno-C 1-6 alkyl), halogen, C 3-8 cycloalkyl, and —CN; in one embodiment, a group selected from the group consisting of halogeno-C 1-6 alkyl and halogen; and in one embodiment, a group selected from the group consisting of trifluoromethyl and fluoro.
  • Examples of the preferable substituents for the “monocyclic aromatic hetero ring which may be substituted” and “bicyclic aromatic hetero ring which may be substituted” in R 2 include, in one embodiment, the substituents described in (a) to (d), (f), (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), —O-(halogeno-C 1-6 alkyl), halogen, C 3-8 cycloalkyl, and —CN; in one embodiment, a group selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen; in one embodiment, a group selected from the group consisting of halogeno-C 1-6 alkyl, —O—(C 1-6 alky
  • cyclic amino which may be substituted with 1 to 5 substituents selected from the group consisting of Group G and oxo, or
  • R 11 is C 1-6 alkyl
  • R 12 is C 1-6 alkyl which may be substituted with 1 to 5 substituents selected from the substituents described in (b) to (o) of Group G, or C 3-8 cycloalkyl which may be substituted with 1 to 5 substituents selected from Group G.
  • cyclic amino which may be substituted with 1 to 5 substituents selected from the group consisting of Group G and oxo, or
  • R 11 is C 1-6 alkyl
  • R 12 is C 1-6 alkyl which may be substituted with 1 to 3 substituents selected from the substituents described in (b) to (g), and (n) of Group G.
  • pyrrolidin-1-yl or piperidin-1-yl in which pyrrolidin-1-yl and piperidin-1-yl are each substituted with 1 to 2 substituents selected from the group consisting of C 1-6 alkyl and halogen-C 1-6 alkyl, or
  • R 11 is C 1-6 alkyl
  • R 12 is C 1-6 alkyl which may be substituted with one group selected from the group consisting of C 3-8 cycloalkyl and —O—(C 1-6 alkyl).
  • R 1 is cyclic amino substituted with 1 to 2 groups selected from the group consisting of C 1-6 alkyl and halogen-C 1-6 alkyl.
  • R 1 is pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl and piperidin-1-yl may be substituted with 1 to 3 substituents selected from Group G.
  • R 1 is pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl and piperidin-1-yl are each substituted with 1 to 2 groups selected from the group consisting of C 1-6 alkyl and halogeno-C 1-6 alkyl.
  • R 1 is pyrrolidin-1-yl substituted with 1 to 2 groups selected from the group consisting of methyl and ethyl.
  • R 1 is —N(—R 11 )(—R 12 ),
  • R 11 is C 1-6 alkyl
  • R 12 is C 1-6 alkyl which may be substituted with a group selected from the group consisting of C 3-8 cycloalkyl and —O—(C 1-6 alkyl),
  • R 1 is —N(—R 11 )(—R 12 ),
  • R 11 is methyl, ethyl, or isopropyl
  • R 12 is methyl, ethyl, isopropyl, isobutyl, cyclopropylmethyl, or methoxyethyl.
  • bicyclic aromatic hetero ring which may be substituted with 1 to 5 substituents selected from Group G.
  • phenyl which may be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), —O-(halogeno-C 1-6 alkyl), halogen, C 3-8 cycloalkyl, and —CN,
  • thienyl which may each be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen,
  • pyridyl which may each be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen, or
  • R 2 is a monocyclic aromatic hetero ring which may be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen.
  • thienyl which may be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen, or
  • pyridyl which may be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, —O—(C 1-6 alkyl), C 3-8 cycloalkyl, and halogen.
  • R 2 is thienyl which may be substituted with 1 to 3 groups selected from the group consisting of C 1-6 alkyl, halogeno-C 1-6 alkyl, C 3-8 cycloalkyl, and halogen.
  • R 2 is thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C 1-6 alkyl and halogen.
  • R 2 is thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of trifluoromethyl and chloro.
  • R 2 is pyridyl which may be substituted with 1 to 3 groups selected from the group consisting of halogeno-C 1-6 alkyl and —O—(C 1-6 alkyl),
  • R 2 is phenyl which may be substituted with 1 to 5 groups selected from the group consisting of C 1-6 alkyl, halogen-C 1-6 alkyl, —O—(C 1-6 alkyl), —O-(halogeno-C 1-6 alkyl), halogen, C 3-8 cycloalkyl, and —CN.
  • R 2 is phenyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C 1-6 alkyl and halogen.
  • thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C 1-6 alkyl and halogen, or
  • phenyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C 1-6 alkyl and halogen.
  • n is an integer of 0 to 4.
  • n is an integer of 0 to 2.
  • R 1 is as described in (1-2) above,
  • R 2 is as described in (2-2) above,
  • R 3 is as described in (3-1) above,
  • n is as described in (5-1) above.
  • R 1 is as described in (1-3) above,
  • R 2 is as described in (2-3) above,
  • n is as described in (5-3) above.
  • R 2 is as described in (2-4) above, and
  • R 1 is as described in (1-6) above,
  • R 2 is as described in (2-14) above, and
  • Examples of the specific compounds included in the present invention include the following compounds or salts thereof:
  • examples of the specific compounds included in the present invention include the following compounds or salts thereof:
  • tautomers or geometrical isomers thereof may exist, depending on the kinds of the substituents.
  • the compound of the formula (I) may be described in only one form of isomers in some cases, but the present invention includes other isomers, isolated forms of the isomers, or a mixture thereof.
  • the compounds of the formula (I) may have asymmetric carbon atoms or asymmetries in some cases, and correspondingly, the optical isomers thereof can exist.
  • the present invention includes the isolated form of the optical isomer of the compound of the formula (I) or a mixture thereof.
  • a pharmaceutically acceptable prodrug of the compound represented by the formula (I) is also included in the present invention.
  • the pharmaceutically acceptable prodrug refers to a compound having a group which can be converted into an amino group, a hydroxyl group, a carboxyl group, or the like, by solvolysis or under a physiological condition.
  • Examples of the groups forming the prodrug include those as described in Prog. Med., 5, 2157-2161 (1985) or “Pharmaceutically Research and Development” (Hirokawn Publishing Company, 1990), vol. 7, Drug Design, 163-198.
  • the salt of the compound of the formula (I) is a pharmaceutically acceptable salt of the compound of the formula (I), and the compounds of the formula (I) may form an acid solution salt or a salt with a base, depending on the kinds of the substituents in some cases.
  • examples thereof include acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid, and with organic acids such as formic acid, acetic acid, propanoic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyl tartaric acid, ditolyl tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, and glutamic acid, and salts with metal anions such as sodium, potassium, magnesium, calcium, and aluminum, and with organic bases such as methylamine, ethylamine, and ethanolamine, salts with various amino acids such as acetyl leucine, lysine,
  • the present invention also includes various hydrates or solvates, and crystal polymorph substances of the compound of the formula (I) and a salt thereof.
  • the present invention also includes the compounds labeled with various radioactive or non-radioactive isotopes.
  • the compound of the formula (I) or a salt thereof can be prepared by applying various know synthetic methods, using the characteristics based on their basic structures or the kinds of the substituents. At this time, depending on the types of the functional groups, it is in some cases effective from the viewpoint of the preparation techniques to protect the functional group with an appropriate protective group (a group which is capable of being easily converted into the functional groups), during the steps from starting materials to intermediates.
  • an appropriate protective group a group which is capable of being easily converted into the functional groups
  • the protective group include the protective groups as described in “Greene's Protective Groups in Organic Synthesis (4th edition, 2006)”, edited by P. G. M. Wuts and T. W. Greene, and the like, which may be appropriately selected and used depending on the reaction conditions. In these methods, a desired compound can be obtained by introducing the protective group to carry out the reaction, and then, if desired, removing the protective group.
  • the prodrug of the compound of the formula (I) can be prepared by introducing a specific group during the steps from starting materials to intermediates, in the same manner as for the above protective groups, or by further carrying out the reaction using the obtained compound of the formula (I).
  • the reaction can be carried out by applying a method known to a person skilled in the art, such as common esterification, amidation, and dehydration.
  • R represents C 1-6 alkyl, which shall apply hereinafter).
  • This reaction is a method for producing a compound of the formula (I) which is a compound of the present invention, by deprotecting a compound of the formula (a).
  • This reaction is carried out using the compound of the formula (a) and a deprotecting reagent in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to heating to reflux, usually for 0.1 hours to 5 days, in a solvent which is insert to the reaction or without a solvent.
  • a solvent which is insert to the reaction or without a solvent.
  • the solvent used herein are not particularly limited, but include alcohols such as methanol, ethanol, n-propanol and the like, N,N-dimethylformamide, tetrahydrofuran, and the like. Further, there are some cases where a mixed solvent of the solvent and water is highly suitable for the reaction.
  • Examples of the deprotecting reagent are not particularly limited, but include bases such as an aqueous sodium hydroxide solution, an aqueous potassium hydroxide solution and the like, and acids such as hydrochloric acid, trifluoroacetic acid and the like.
  • This production process is a method for producing the compound of the formula (a) which is a starting material of the compound of the formula (I).
  • examples of L 1 include chloro and the like.
  • This step is a step of preparing a compound of the formula (d) by subjecting a compound of the formula (b) and a compound of the formula (c) to an amidation reaction.
  • the reaction is carried out using the formula (b) and the compound of the formula (c) in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to under heating, preferably at ⁇ 20° C. to 60° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction, in the presence of a condensing agent.
  • solvent used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane, cyclopentylmethyl ether and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, water, and a mixture thereof.
  • aromatic hydrocarbons such as benzene, toluene, xylene and the like
  • halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like
  • ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxye
  • condensing reagent examples include 1-(3-dimethylamino propyl)-3-ethylcarbodiimide or a hydrochloride thereof, dicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole, diphenylphosphoric azide, phosphorous oxychloride, N-[( ⁇ [(1Z)-1-cyano-2-ethoxy-2-oxoethylidene]amino ⁇ oxy)morpholin-4-yl)methylene]-N-methylmethanaminium hexafluorophosphate (COMU), and the like, but are not limited thereto.
  • COMP 1-(3-dimethylamino propyl)-3-ethylcarbodiimide or a hydrochloride thereof, dicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole, diphenylphosphoric azide, phosphorous oxychloride, N-[(
  • an additive for example, 1-hydroxybenzotriazole
  • an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like
  • an inorganic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
  • the carboxylic acid (c) is converted to a reactive derivative thereof, and then the reactive derivative is reacted with the amine (b) can also be used.
  • the reactive derivative of the carboxylic acid include acid halides obtained by the reaction with a halogenating agent such as phosphorus oxychloride, thionyl chloride or the like, mixed acid anhydrides obtained by the reaction with isobutyl chloroformate or the like, and active esters obtained by condensation with 1-hydroxybenzotriazole or the like.
  • the reaction of these reactive derivatives and the compound (b) can be carried out under the temperature condition ranging from under cooling to under heating, preferably at ⁇ 20° C. to 60° C., in a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers and the like.
  • This step is a step of preparing a compound of the formula (f) by reacting a compound of the formula (d) with a compound of the formula (e).
  • This reaction is carried out using the formula (d) and the compound of the formula (e) in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to under heating to reflux, preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent.
  • solvent used herein examples include aromatic hydrocarbons such as benzene, toluene, xylene and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.
  • aromatic hydrocarbons such as benzene, toluene, xylene and the like
  • ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like
  • halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,
  • an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like, or an organic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
  • This step is a step of preparing a compound of the formula (g) by introducing an acetoxymethyl group into the 5-position of thiazole in the compound of the formula (f).
  • the compound of the formula (f) is reacted with an aqueous formaldehyde solution or paraformaldehyde in the presence of an acetic acid solvent, which can be carried out under the temperature condition ranging from at room temperature to under heating to reflux.
  • the reaction can also be carried out by adding acetic acid into a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers and the like, instead of the acetic acid solvent.
  • the reaction can also be carried out by further adding acetic anhydride.
  • This step is a step of preparing a compound of the formula (a) by reacting a compound of the formula (g) with a compound of the formula (h) under a basic condition.
  • the present reaction can be carried out by reacting the compound of the formula (g) with the compound of the formula (h) in the presence of an organic base such as triethylamine and N,N-diisopropylethylamine, in an organic solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers, esters, acetonitrile, N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone and the like.
  • the compound of the formula (h) may also used in an excess amount instead of the organic base.
  • the reaction can be carried out under the temperature condition ranging from under cooling to at room temperature; from at room temperature to under heating; or from at room temperature to under refluxing.
  • the compound of the formula (a) can be directly obtained while not isolating the compound of the formula (g) by adding the compound of the formula (h) into the reaction mixture of Step 3.
  • This production process is another preparation method for the compound of the formula (a), which is a starting material of the compound of the formula (I).
  • the protective groups represented by P 1 and P 2 the groups of amino groups described in “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4 th edition, John Wiley & Sons Inc., 2006, and the like can be used.
  • the P 1 include acetyl, trifluoroacetyl and the like
  • examples of P 2 include t-butoxycarbonyl and the like
  • examples of L 2 include bromo and the like.
  • This compound is a step of protecting the amino group of the compound (b).
  • the present reaction can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4 th edition, John Wiley & Sons Inc., 2006.
  • This step is a step of preparing a compound of the formula (k) by introducing an acetoxymethyl group into the 5-position of thiazole in a compound of the formula (j).
  • the reaction conditions are the same as in Step 3 of Production Process 2.
  • the step is a step of preparing a compound of the formula (m) by reacting a compound of the formula (h) and a compound of the formula (k) under a basic condition.
  • the reaction conditions are the same as in Step 4 of Production Process 2.
  • This step is a step of deprotecting a protective group P 1 of an amino group of the compound (m).
  • the present reaction can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4 th edition, John Wiley & Sons Inc., 2006.
  • This step is a step of obtaining a compound of the formula (q) by subjecting a compound of the formula (o) and a compound of the formula (p) to an amidation reaction.
  • the reaction conditions are the same as in Step 1 of Production Process 2.
  • This step is a step of preparing a compound of the formula (s) by reacting a compound of the formula (q) with a compound of the formula (r).
  • the reaction conditions are the same as in Step 2 of Production Process 2.
  • This step is a step of deprotecting a protective group P 2 of a compound of the formula (s).
  • This step can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4 th edition, John Wiley & Sons Inc., 2006”.
  • This step is a step of obtaining the compound of the formula (a) by reacting a compound of the formula (t) and a compound of the formula (n).
  • the present reaction is carried out using the compound (t) and the compound (u) in equivalent amounts, or either thereof in an excess amount, and stirring the mixture under the temperature condition ranging from under cooling to under heating to reflux, preferably at 0° C. to 100° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction, or without a solvent.
  • solvent used herein examples include aromatic hydrocarbons such as benzene, toluene, xylene and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone, ethyl acetate, acetonitrile, and a mixture thereof.
  • aromatic hydrocarbons such as benzene, toluene, xylene and the like
  • ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like
  • halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,
  • an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like
  • an inorganic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
  • the compound of the formula (I) is isolated and purified as its free compound, or a salt, a hydrate, a solvate, or crystal polymorph substance thereof.
  • the salt of the compound of the formula (I) can also be prepared by a conventional method.
  • Isolation and purification are carried out by employing general chemical operations such as extraction, fractional crystallization, and various types of fractional chromatography.
  • Various isomers can be prepared by selecting appropriate starting compound, or separated by separation using differences in the physicochemical properties among the isomers.
  • the optical isomers can be obtained by means of general optical resolution methods of racemic compounds (for example, fractional crystallization introducing the compound into a diastereomer salt with an optically active base or acid; chromatography using a chiral column or the like; and others), or can also be prepared from appropriate optically active starting compound.
  • a human muscarinic M 3 receptor gene (GenBank Accession No.: NM_00740.2) was introduced into an expression vector pcDNA3.1TM (Life Technologies).
  • a vector expressing a human muscarinic M 3 receptor was introduced into a CHO—K1 cell (ATCC No.: CCL-61). The introduction was carried out according to the attached instructions, using a transfection reagent, Lipofectoamine (registered trademark) 2000 Reagent (Life Technologies). The cells were incubated in an alpha Modified Eagle Minimum Essential Medium ( ⁇ -MEM) including 2 mM glutamine, 10% fetal bovine serum, and 2.0 mg/mL. Geneticin (registered trademark) (Life Technologies) for 4 weeks to acquire a drug-resistant clone.
  • ⁇ -MEM alpha Modified Eagle Minimum Essential Medium
  • Geneticin registered trademark
  • the cells obtained in b) above were suspended in an ⁇ -MEM including 2 mM glutamine, 10% fetal bovine serum, and 0.2 mg/mL. Geneticin (registered trademark) to the amount from 1.2 to 1.5 ⁇ 10 4 cells/well the day before the experiment, dispersed into a 384-well plate (Model No. 355962, BD Biosciences), and incubated overnight at 37° C. and 5% CO 2 .
  • the medium was replaced with a loading buffer (an assay buffer (Hank's balanced salt solution (HBSS), 1 g/L BSA, 20 mM HEPES (pH 7.5), and 2.5 mM probenecid), including 3.1 ⁇ M Fluo 4-AM (Dojindo Laboratories) and incubated for about 2 hours at room temperature. Thereafter, the cells were washed with a plate washer EL ⁇ 405TM (BIO-TEK Instrument, Inc.) set with the assay buffer, and set in an intracellular Ca 2+ concentration measuring system (FLIPR intra (registered trademark), Molecular Device Co.).
  • test substances final concentration of 1 ⁇ M or 10 ⁇ M
  • carbachol Sigma, final concentration of 0.0024 nM to 10 ⁇ M
  • the test substances were added to the cells in the device and after about 5 minutes, carbachol was added to the cells.
  • An increase rate of the intracellular Ca 2+ concentration by carbachol was measured (excitement wavelength of 470 nm to 495 nm and a fluorescence wavelength of 515 nm to 575 nm).
  • a shift toward a lower concentration side of a carbachol concentration-response curve by the test substance was used as an index. That is, a minimum value in the carbachol response was taken as 0%; a minimum value in the carbachol response was taken as 100% from the concentration-response curve of carbachol; the carbachol concentration exhibiting a 50% response was calculated as an EC 50 value, using a Sigmoid-Emax model non-linear regression method, and thus, the muscarinic M 3 receptor-positive allosteric modulator activity was determined by dividing the EC 50 value of carbachol in the absence of the test substance by the EC 50 value of carbachol in the presence of the test substance.
  • the value of the muscarinic M 3 receptor-positive allosteric modulator activity becomes 10, showing that the test substance causes a 10-fold shift in the EC 50 value toward to the low concentration side.
  • the columns of 10 ⁇ M (-fold shift) show the values in a case where the test substance is added to a final concentration of 10 ⁇ M and the columns of 1 ⁇ M (-fold shift) show the values in a case where the test substance is added to a final concentration of 1 ⁇ M.
  • the human c-Mpl-introduced Ba/F3 cell proliferation action was measured by the following method.
  • a human c-Mpl receptor gene (GenBank Accession No.: M90102.1) was transfected into an expression vector pEF-BOS (Nucleic Acids Research, 18; pp 4322 (1990)).
  • a vector expressing a human c-Mpl receptor was introduced into a Ba/F3 cell (RIKEN BRC: RCB0805).
  • an electroporation method was used.
  • pEF-BOS-c-mpl (10 ⁇ g), pSV2bsr (1 ⁇ g, Kaken Pharmaceutical Co., Ltd.) and 1 ⁇ 10 9 of Ba/F3 cells were put into cuvettes with a gap width of 0.4 cm and electroporated under a condition of 1.5 kV (25 ⁇ F) in a Gene Pulser (registered trademark) (BioRad).
  • the cells were incubated in an RPMI-1640 medium supplemented with a 0.5% WEHI conditioned medium (BD Biosciences) and 10% fetal bovine serum for 3 days, and thereafter, and the cells were incubated for 30 days in an RPMI-1640 medium, to which 10 ⁇ g/mL blasticidin had been further added, thereby acquiring a drug-resistance clone.
  • RPMI-1640 medium supplemented with a 0.5% WEHI conditioned medium (BD Biosciences) and 10% fetal bovine serum for 3 days, and thereafter, and the cells were incubated for 30 days in an RPMI-1640 medium, to which 10 ⁇ g/mL blasticidin had been further added, thereby acquiring a drug-resistance clone.
  • the cells obtained in b) above were dispersed into an RPMI-1640 medium supplemented with a 0.5% WEHI conditioned medium and 10% fetal bovine serum, and used.
  • the test substances final concentration of 100 mM to 10 ⁇ M
  • a medium for assay an RPMI-1640 medium supplemented with 10% fetal bovine serum
  • the cells after the medium had been replaced with the medium for assay were dispensed to a 384-well plate to which the test substance had been added, to 1 ⁇ 10 4 cells/well, and incubated overnight at 37° C. and 5% CO 2 .
  • a cell proliferation rate (%) was calculated from the absorbance of the well to which the test substance has been added. From the obtained results, the test substance concentration exhibiting 30% proliferation by a Sigmoid-Emax model non-linear regression method was calculated as an EC 30 value.
  • Test Example 2 10 ⁇ M 1 ⁇ M EC 30 Ex (-fold shift) (-fold shift) (nM) 3 253 101 780 4 200 25 >3000 10 87 21 >10000 11 226 33 >10000 12 178 33 >10000 13 326 43 >10000 15 159 31 >10000 17 109 15 >10000 21 149 25 >10000 27 330 31 >10000 28 108 36 5300 33 182 40 >10000 34 116 18 >10000 41 160 43 >10000 42 141 39 >10000 43 224 76 >10000 46 199 29 >10000 48 113 27 >10000 49 224 67 >10000 50 190 108 2300 51 287 102 2600 52 196 36 >10000 54 134 36 >10000 60 235 33 9700 61 229 35 1300 62 195 37 >10000 63 186 39 >10000 64 128 23 >10000 65 90 24 >10000 67 114 40 >10000 69 177 27 >10000
  • Test Example 2 10 ⁇ M 1 ⁇ M EC 30 Ex (-fold shift) (-fold shift) (nM) 71 151 28 >10000 72 152 31 >10000 79 171 60 >10000 81 94 89 1800 82 43 11 500 91 139 19 >10000 92 203 30 >10000 95 233 91 >10000 97 121 55 3000 100 229 82 2800 101 112 64 3200 103 307 202 2700 104 195 75 1700 106 270 41 >10000 107 318 73 >10000 108 169 56 >10000 109 191 30 >10000 111 627 203 5000 118 167 57 >10000 119 503 110 >10000 124 101 28 >10000 126 318 79 >10000 128 192 73 8000 129 67 >10000 130 151 95 >10000 132 41 15 >10000 133 164 30 >10000 135 204 25 >10000 140 158 28 >
  • Example 1 a substantial number of the Example compounds which had been subjected to the present test shifted the EC 50 values to almost 100-fold or more toward a lower concentration side when added at 10 ⁇ M, and shifted the EC 50 values to almost 10-fold or more toward a lower concentration side when added at 1 ⁇ M.
  • Example compounds of the present invention from the viewpoint that the compounds alone do not change the intracellular Ca 2+ concentration, it was found that these compounds have no muscarinic M 3 receptor agonistic activity.
  • Test Example 2 it was found that a substantial number of the Example compounds which had been subjected to the present test have a weak human c-Mpl-introduced Ba/F3 cell proliferative activity or have none.
  • the compound of the present invention is used as an agent for preventing or treating bladder/urinary tract diseases associated with bladder concentrations via a muscarinic M 3 receptor, as a muscarinic M 3 receptor-positive allosteric modulator, and thus preferably has a weak or none increased platelet action based on c-Mpl-introduced Ba/F3 cell proliferative activity.
  • Table 1 of Patent Document 1 above discloses that the compound of Example 315 represented by the formula (A1) above has 3.2 nM of EC 30 value of c-Mpl-introduced Ba/F3 cell proliferation action.
  • the effect of the Example compounds of the present invention in the electrical field stimulation-induced contraction of the rat isolated bladder was measured by the following method. That is, a bladder specimen having a width of about 2 mm and a length of about 10 mm in the longitudinal direction from the bladder isolated from a Sprague-Dawley (SD) female rat (Japan SLC, Inc.) was prepared. The prepared bladder specimen was suspended in an organ bath filled with 10 mL of a Krebs-Henseleite solution. The Krebs-Henseleite solution was aerated at 95% 02 and 5% CO 2 , and kept at 37° C.
  • the contraction was caused twice with 60 nM KCl.
  • the concentration was caused by carrying out electrical field stimulation at 20 V with an electrical stimulation device (Nihon Kohden) (a stimulation frequency of 8 Hz, a pulse width of 0.3 msec, and a stimulation time of 10 seconds).
  • an electrical stimulation device Nihon Kohden
  • a voltage was adjusted to obtain a contraction height of approximately 50% of the contractile response at 20 V.
  • test substances dissolved in 100% dimethyl sulfoxide in advance (final concentrations of 3 ⁇ M, 10 ⁇ M, and 30 ⁇ M) was added thereto.
  • the test substances were cumulatively administered at the following concentrations after the low-concentration contractile response had been stabilized.
  • the response was taken into a personal computer through a PowerLab (registered trademark) (AD Instruments, Inc.), and analyzed by LabChart (registered trademark) (AD Instruments, Inc.).
  • AUC area under curve, AUC
  • Example compounds which have been subjected to the present test do not cause contraction in a state in which there is no electrical stimulation and the compounds alone do not show a bladder contraction action.
  • Example compounds alone which have been subjected to the present test, do not cause a contraction action in the isolated rate bladder, but have an action of enhancing electrical field stimulation-induced contraction.
  • Example compounds of the present invention in the pelvic nerve electrical stimulation-induced elevation of intravesical pressure using rats as an action of nerve stimulation-dependent bladder contraction in vivo was measured by the following method. That is, SD female rats (Japan SLC, Inc.) were used and its lower abdomen was dissected at the midline under pentobarbital anesthesia (50 mg/kg ip). After ligating and cutting the ureter on both sides, a cannula (PE-5) for measuring the intravesical pressure was inserted into the bladder from the external urethral opening and fixed b a clip.
  • PE-5 cannula
  • the other side was connected to a pressure transducer to measure the intravesical pressure.
  • the pelvic nerve in the vicinity of the bladder was peeled and an electrode for nerve stimulation (unique Medical) was placed.
  • the abdominal cavity was filled with mineral oil (MP BIOMEDICALS).
  • electrical stimulation stimulation voltage: 10 V, stimulation frequency: 8 Hz, pulse width: 0.3 msec, and stimulation time: 10 seconds
  • stimulation voltage stimulation voltage: 10 V
  • stimulation frequency 8 Hz
  • pulse width 0.3 msec
  • stimulation time 10 seconds
  • the voltage was adjusted to elicit about 50% to 70% elevation of intravesical pressure elicited at 10 V.
  • the increase in the intravesical pressure by electrical stimulation was stabilized three times or more, and the test substance (an administration amount of 3 mg/kg) was then administered from the catheter detained in the vein at a volume of 1 mL/kg, thus measuring an effect of the elevation of the intravesical pressure of the test substance for 1 hour.
  • the test substance was dissolved in water supplemented with 10% dimethylsulfoxide and 10% Cremophor.
  • the response was applied to a personal computer through a PowerLab (registered trademark) and analyzed by Lab-Chart (registered trademark).
  • the AUC of each elevation of the intravesical pressure was calculated, the intravesical pressure elevation rate (% of pre) after the treatment with the test substance was calculated by taking an average value of the values measured three times before the treatment with the test substance as 100%, and the maximum effect during a period within one hour after administration of the compound was considered as the effect of the test substance.
  • Example compounds evaluated in the present test do not cause an elevation of the intravesical pressure in a state in which electrical stimulation is not given, and the compounds alone do not show elevation of the intravesical pressure.
  • Example compounds listed in Table 4 alone do not show elevation of the intravesical pressure but have an action of enhancing effect on the pelvic nerve electrical stimulation-induced elevation of intravesical pressure in the anesthetized rats.
  • the compound of the formula (I) has a muscarinic M 3 receptor-positive allosteric modulator activity, and further, it enhances the bladder contraction in a nerve stimulation-dependent manner in in vitro, as well as enhances an elevation in the intravesical pressure in a nerve stimulation-dependent manner in in vitro. Accordingly, the compound of the formula (I) can be used to prevent or treat bladder/urinary tract diseases associated with bladder contractions via a muscarinic M 3 receptor, in particular, voiding dysfunction or urine storage dysfunction in the bladder/urethral diseases.
  • the compound of the formula (I) can be used for preventing or treating, for example, voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, neurogenic bladder, urethra relaxation failure, detrusor-external urethral sphincter dyssynergia, overactive bladder, urinary frequency, nocturia, urinary incontinence, benign prostatic hyperplasia, interstitial cystitis, chronic prostatitis, and urinary tract stones.
  • the compound of the formula (I) can be used for preventing or treating voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, and neurogenic bladder.
  • the compound of formula (I) can become a therapeutic drug that is more excellent in safety from the viewpoint that the compound alone does not show an agonistic effect on a muscarinic M 3 receptor, but shows an effect on enhancing the nerve stimulation-dependent bladder contraction, and accordingly, cholinergic side effects that have been reported in the existing drugs can be avoided.
  • a pharmaceutical composition including one or two or more kinds of the compound of the formula (I) as an active ingredient can be prepared using an excipient which is usually used in the art, that is, an excipient for a pharmaceutical preparation, a carrier for a pharmaceutical preparation, and the like, according to a method usually used.
  • Administration can be accomplished either by oral administration via tablets, pills, capsules, granules, powders, solutions, and the like, or parenteral administration via injections, such as intraarticular, intravenous, and intramuscular injections, suppositories, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalers, and the like.
  • parenteral administration via injections, such as intraarticular, intravenous, and intramuscular injections, suppositories, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalers, and the like.
  • a solid composition for oral administration tablets, powders, granules, and the like are used.
  • one kind or two or more kinds of the active ingredients are mixed with at least one inactive excipient.
  • the composition may contain inactive additives such as a lubricant, a disintegrating agent, a stabilizer, or a solubilization assisting agent. If necessary, tablets or pills may be coated with a sugar or with a film of a gastric or enteric coating substance.
  • the liquid composition for oral administration includes pharmaceutically acceptable emulsions, solutions, suspensions, syrups elixirs, or the like, and also includes generally used inert diluents, for example, purified water or ethanol.
  • the liquid composition may also include auxiliary agents such as a solubilization assisting agent, a moisturizing agent, and a suspending agent, sweeteners, flavors, aromatics, and antiseptics, in addition to the inert diluent.
  • the injections for parenteral administration include sterile aqueous or non-aqueous solution preparations, suspensions, or emulsions.
  • the aqueous solvent includes, for example, distilled water for injection and saline.
  • the non-aqueous solvent include alcohols such as ethanol.
  • Such a composition may further include a tonicity agent, an antiseptic, a moistening agent, an emulsifying agent, a dispersing agent, a stabilizing agent, or a solubilizing assisting agent. These are stabilized, for example, by filtration through a bacteria retaining filter, blending of a bactericide, or irradiation. In addition, these can also be used by preparing a sterile solid composition, and dissolving or suspending it in sterile water or a sterile solvent for injection prior to its use.
  • agent for external use examples include ointments, hard plasters, creams, jellies, cataplasms, sprays, and lotions.
  • the agent further contains generally used ointment bases, lotion bases, aqueous or non-aqueous liquid preparations, suspensions, emulsions, or the like.
  • transmucosal agents such as an inhaler and a transnasal agent
  • those in the form of a solid, liquid, or semi-solid state are used, and can be prepared in accordance with a method known in the related art.
  • a known excipient and also a pH adjusting agent, an antiseptic, a surfactant, a lubricant, a stabilizing agent, a thickening agent, or the like may be appropriately added thereto.
  • an appropriate device for inhalation or blowing can be used.
  • a compound may be administered alone or as a powder of formulated mixture, or as a solution or suspension in combination with a pharmaceutically acceptable carrier, using a known device or sprayer such as a metered administration inhalation device.
  • a dry powder inhaler or the like may be for single or multiple administration use, and a dry powder or a powder-containing capsule may be used.
  • this may be in a form such as a pressurized aerosol spray that uses an appropriate propellant agent, for example, a suitable gas such as chlorofluoroalkanes, and carbon dioxide, or other forms.
  • the daily dose is from about 0.001 mg/kg to 100 mg/kg, preferably from 0.1 mg/kg to 30 mg/kg, and more preferably from 0.1 mg/kg to 10 mg/kg, per body weight, administered in one portion or in 2 to 4 divided portions.
  • the daily dose is suitably administered from about 0.0001 mg/kg to 10 mg/kg per body weight, once a day or two or more times a day.
  • a transmucosal agent is administered at a dose from about 0.001 mg/kg to 100 mg/kg per body weight, once or plural times a day. The dose is appropriately decided in response to the individual case by taking the symptoms, the age, and the gender, and the like into consideration.
  • a pharmaceutical composition of the present invention comprises 0.001% by weight to 100% by weight of, as an embodiment, 0.01% by weight to 50% by weight of, one or more of the compound of the formula (I) or a salt thereof which is the active ingredient.
  • the compound of the formula (I) may be used in combination with various agents for treating or preventing diseases on which the compound of the formula (I) is considered to show the effect. Such combined preparations may be administered simultaneously, or separately and continuously, or at a desired time interval.
  • the preparations to be co-administered may be a blend, or may be prepared individually.
  • the production process for the compound of the formula (I) will be described in more detail with reference to Examples. Further, the present invention is not limited to the compounds described in the Examples below. Further, the production processes for the starting compounds will be described in Preparation Examples. In addition, the production processes for the compound of the formula (I) are not limited to the production processes of the specific Examples shown below, but the compound of the formula (I) can be prepared by a combination of these production processes or a method that is apparent to a person skilled in the art.
  • nomenclature software such as ACDC/Name (registered trademark, Advanced Chemistry Development, Inc.) may be used for nomenclature of compounds in some cases.
  • the powder X-ray diffraction is measured using RINT-TTRII under the condition of a tube: Cu, a tube current: 300 mA, a tube voltage: 50 kV, a sampling width: 0.020°, a scanning speed: 4°/min, a wavelength: 1.54056 angstroms, and a measurement diffraction angle (2 ⁇ ): 2.5° to 40°. Further, a device including data processing was handled in accordance with the method and procedure instructed in each device.
  • the values obtained from various spectra may cause some errors according to the direction of the crystal growth, particle sizes, measurement conditions, and the like in some cases. Accordingly, considering these errors, in the present specification, the description of diffraction angles (2 ⁇ (°)) in the powder X-ray diffraction patterns is measured value, but depending on the measuring conditions, these diffraction angles mean that error ranges which are usually acceptable may occur and means that they are approximate values. Usually, the error range of the diffraction angle (2 ⁇ (°)) in the powder X-ray diffraction is ⁇ 0.2°.
  • crystal lattice spacing and general patterns are important in the certification of crystal identity, and the diffraction angle and the diffraction intensity may vary slightly depending on the direction of crystal growth, the particle size, and the measurement condition, and they should not be strictly construed.
  • HCl in the structural formula indicates that the compound is a monohydrochloride; 2HCl indicates that the compound is a dihydrochloride: 3HCl indicates that the compound is a trihydrochloride, and 2 maleic acid indicates that the compound is a dimalate dimaleate.
  • a concentration of mol/L is represented by M.
  • a 1 M aqueous sodium hydroxide solution means a 1 mol/L aqueous sodium hydroxide solution.
  • N-[4-(4-Chlorothiophen-2-yl)-1,3-thiazol-2-yl]-2,2,2-trifluoroacetamide (5.0 mL), and a 36% aqueous formaldehyde solution (2.5 mL) were mixed, followed by stirring at 60° C. for 1 hour.
  • the reaction mixture was concentrated under reduced pressure and diluted with ethyl acetate.
  • the mixture was washed with a saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure.
  • the obtained compound and ethanol (625 mL) were mixed, and thiourea (35 g) was added thereof, followed by stirring at 65° C. to 75° C. for 2 hours.
  • the reaction mixture was ice-cooled, and water (625 mL) was added thereto.
  • the the mixture was added a 1 M sodium hydroxide (600 mL), followed by stirring for 30 minutes.
  • the solid was collected by filtration, and ethanol (30% aqueous, 600 mL) was added thereto and dissolved at 76° C.
  • the obtained solution was cooled to room temperature and stirred overnight.
  • reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (50 mL) and water (100 mL) were added thereto, followed by extraction with chloroform.
  • the organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure.
  • the obtained solid was mixed with hexane (20 mL) and diethyl ether (4 mL), and the solid was collected by filtration to obtain sodium 3-(4- ⁇ 5-[(4-[3-cyano-5-(trifluoromethyl)phenyl]-5- ⁇ [(2S)-2-isopropylpyrrolidin-1-yl]methyl ⁇ -1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl ⁇ piperazin-1-yl)propanoate (149 mg) as a solid.
  • the reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with water and saturated brine.
  • the organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure.
  • the residue was purified by basic silica gel column chromatography (hexane-ethyl acetate).
  • the obtained solid was mixed with hexane (10 mL) and diethyl ether (2 mL), and the solid was collected by filtration to obtain sodium 3-(4- ⁇ 5-[(5- ⁇ [(2R)-2-methylpiperidin-1-yl]methyl ⁇ -4-[3-methyl]-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazine-2-yl ⁇ piperazin-1-yl)propanoate (284 mg) as a solid.
  • the residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) and purified by silica gel column chromatography (hexane-ethyl acetate).
  • the obtained compound was mixed with ethanol (2 mL) and tetrahydrofuran (2 mL), and a 1 M aqueous sodium hydroxide solution (1 mL) was added thereto, followed by stirring at room temperature for 1 hour.
  • To the reaction mixture was added 1 M hydrochloric acid (1 mL) and water, the mixture was extracted with chloroform/isopropanol, and the organic layer was washed with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure.
  • the crystals obtained in the present Examples have peaks of powder X-ray diffraction at 2 ⁇ (°) 5.7, 6.6, 10.5, 12.0, 13.3, 15.8, 16.6, 17.3, 19.0, and 26.2.
  • the compound of the formula (I) or a salt thereof is a muscarinic M 3 receptor-positive allosteric modulator, and can thus be used as an agent for preventing or treating bladder/urinary tract diseases associated wit bladder contractions via a muscarinic M 3 receptor.

Abstract

[Problem]To provide a compound which is useful as an active ingredient for a pharmaceutical composition for preventing or treating urine storage dysfunction, voiding dysfunction, lower urinary tract dysfunction, and the like.[Means for Solution]The present inventors have found that a thiazole derivative substituted with pyrazinylcarbonylamino at the 2-position is an excellent muscarinic M3 receptor-positive allosteric modulator and is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, thereby completing the present invention. 2-Acylaminothiazole derivative or a salt thereof of the present invention is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, for example voiding dysfunction such as underactive bladder.

Description

CROSS REFERENCE TO RELATED APPLICATION
This application is a National Stage entry under 35 USC 371 of PCT/JP2015/066321, filed on Jun. 5, 2015, and claims priority to Japanese Patent Application No. 2014-118046, filed on Jun. 6, 2014.
TECHNICAL FIELD
The present invention relates to a 2-acylaminothiazole derivative or a salt thereof which is useful as an active ingredient for a pharmaceutical composition, in particular, a pharmaceutical composition for treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M3 receptor.
BACKGROUND ART
The important roles of the lower urinary tract are urine storage and voiding, which are regulated by a coordinated action of the bladder and the urethra. That is, during urine storage, the bladder smooth muscle is relaxed and the urethral sphincter is contracted, whereby a state in which urethral resistance is high is maintained and urinary continence is maintained. On the other hand, during voiding, the bladder smooth muscle is contracted, the urethra smooth muscle is relaxed, and contraction of the external urethral sphincter is also inhibited. Examples of the lower urinary tract disorder include urine storage dysfunction such as overactive bladder, in which urine cannot be retained during urine storage, and voiding dysfunction, in which urine cannot be drained sufficiently during voiding due to an increase in the urethral resistance or a decrease in the bladder contractile force. These two disorders may develop simultaneously in some cases.
Voiding dysfunction is caused by a decrease in the bladder contractile force or an increase in urethral resistance during voiding, and causes difficulty in voiding, straining during voiding, a weak urine stream, extension of voiding time, an increase in residual urine, a decrease in voiding efficiency, or the like. The decrease in the bladder contractile force during voiding is referred to as underactive bladder, acontractile bladder, or the like. As a factor causing such a decrease in the bladder contractile force during voiding, for example, aging, diabetes mellitus, benign prostatic hyperplasia, neurological diseases such as Parkinson's disease and multiple sclerosis, spinal cord injury, neurological disorders by pelvic surgery, and the like have been known (Reviews in Urology, 15; pp. 11-22 (2013)).
As a mechanism to cause bladder contraction during voiding, involvement of muscarinic receptor stimulation has been known. That is, during urination, the pelvic nerve which is a parasympathetic nerve governing the bladder is excited to release acetylcholine from nerve terminals. The released acetylcholine binds to a muscarinic receptor present in the bladder smooth muscle to cause contraction of the bladder smooth muscle (Journal of Pharmacological Sciences, 112; pp. 121-127 (2010)). The muscarinic receptors are currently classified into five subtypes, M1, M2, M3, M4, and M5, and it has been known that the subtypes involving the contraction in the bladder smooth muscle is mainly M3 (Pharmacological Reviews, 50; pp. 279-290 (1998); The Journal of Neuroscience, 22; pp. 10627-10632 (2002)).
As a therapeutic drug for a decrease in bladder contractile force during voiding, bethanechol chloride which is a non-selective muscarinic receptor agonist and distigmine bromide which is a cholinesterase inhibitor have been known. However, it has been known that these drugs have cholinergic side effects such as diarrhea, abdominal pain, and perspiration. In addition, there may be cases where cholinergic crisis is occurred as a serious side effect, which require attention during use (Uhretid (registered trademark), tablet 5 mg, package insert, Torii Pharmaceutical Co., Ltd., and Besacholine (registered trademark) powder 5%, package insert, Eisai Co., Ltd.).
On the other hand, as a cause of an increase in urethral resistance, voiding dysfunction associated with benign prostatic hyperplasia has been well-known, which is characterized in that the urethra is partially occluded by nodular enlargement of the prostatic tissue. Currently, an adrenergic a, receptor antagonist has been used as a therapeutic drug for voiding dysfunction associated with benign prostatic hyperplasia (Pharmacology, 65; pp. 119-128 (2002)). On the other hand, the effectiveness of the adrenaline α, receptor antagonist for voiding dysfunction that is not associated with benign prostatic hyperplasia is unclear, as compared with the effectiveness against voiding dysfunction that is associated with benign prostatic hyperplasia (Journal of Pharmacological Sciences, 112; pp. 121-127 (2010)).
Furthermore, for voiding dysfunction caused by a decrease in bladder contractile force or an increase in urethral resistance, residual urine after voiding may be observed in some cases. The increased residual urine may cause a decrease in effective bladder capacity, and thus cause overactive bladder symptoms such as urinary frequency or severe symptoms such as hydronephrosis in some cases.
There has been a demand for a more effective therapeutic drug for such bladder/urethral diseases due to a decrease in the bladder contractile force or an increase in urethral resistance during voiding, or symptoms thereof (Reviews in Urology, 15; pp. 11-22 (2013)).
Patent Document 1 discloses that a compound represented by the following general formula (A) including a compound of the formula (A1) below, which is disclosed in Example 315, has a Ba/F3 cell proliferative activity through a human c-mycloproliferative leukemia virus type P (c-Mpl), and has thrombocyte increasing activity.
Figure USRE049111-20220621-C00001
(in which R3 represents an aromatic hetero ring which may be substituted, or the like. For the other symbols, refer to the patent publication).
Patent Document 2 discloses that a compound represented by the following general formula (B) has an AMPK pathway activating action.
Figure USRE049111-20220621-C00002
(in which Ring B represents a heteroarylene or the like, J represents —NR13C(O)— or the like, D1, D2 and D3 each represent N, CH, or the like, E represents —NR1R2 or the like, and R1 and R2 may be combined with an adjacent nitrogen atom to form a heterocycloalkyl which may be substituted. For the other symbols, refer to this publication).
Non-Patent Document 1 discloses that a compound represented by the following formula (C1) is an allosteric enhancer of a muscarinic M3 receptor.
Figure USRE049111-20220621-C00003
Non-Patent Document 2 discloses that WIN 62,577 represented by the following formula is a rat NK1 receptor antagonist and, at the same time, an allosteric enhancer of a muscarinic receptor.
Figure USRE049111-20220621-C00004
RELATED ART Patent Document
  • [Patent Document 1] WO 2005/007651
  • [Patent Document 2] WO 2012/016217
  • [Non-Patent Document 1] Molecular Pharmacology, 55; pp 778-786 (1999)
  • [Non-Patent Document 2] Molecular Pharmacology, 62; pp 1492-1505 (2002)
DISCLOSURE OF INVENTION Problems to be Solved by the Invention
The present invention provides a novel compound which is expected as an active ingredient for a pharmaceutical composition, in particular, for a pharmaceutical composition for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, which acts as a muscarinic M3 receptor-positive allosteric modulator.
Means for Solving the Problems
The present inventors have found that a thiazole derivative substituted with pyrazinylcarbonylamino at the 2-position is an excellent muscarinic M3 receptor-positive allosteric modulator and is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, thereby completing the present invention.
That is, the present invention relates to a compound of the formula (I) or a salt thereof, and a pharmaceutical composition comprising a compound of the formula (I) or a salt thereof and an excipient.
Figure USRE049111-20220621-C00005
(wherein
R1 is —N(—R11)(—R12), or cyclic amino which may be substituted,
R11 is C1-6 alkyl,
R12 is C1-6 alkyl which may be substituted, or C3-8 cycloalkyl which may be substituted,
R2 is aryl which may be substituted, monocyclic aromatic hetero ring which may be substituted, or bicyclic aromatic hetero ring which may be substituted,
R3's are the same as or different from each other, and are each C1-6 alkyl,
W is C1-6 alkylene, and
n is an integer of 0 to 4).
Further, unless specifically described otherwise, when symbols in one formula in the present specification are also used in other formulae, same symbols denote same meanings.
Further, Patent Document 1 does not disclose a specific compound which is a compound of the formula (A) wherein R3 is pyrazinyl, and neither discloses nor suggests an action on a muscarinic receptor or an action on bladder/urethral diseases.
Furthermore, Patent Document 2 does not disclose a specific compound which is a compound of the formula (B) wherein ring B is thiazole, and neither discloses nor suggests an action on a muscarinic receptor or an action on bladder/urethral diseases.
Further, the present invention relates to a pharmaceutical composition comprising the compound of the formula (I) or a salt thereof, and a pharmaceutically acceptable excipient. Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, comprising the compound of the formula (I) or a salt thereof. Furthermore, the present invention relates to an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, comprising the compound of the formula (I) or a salt thereof.
Moreover, the present invention relates to use of the compound of the formula (I) or a salt thereof for the manufacture of a pharmaceutical composition for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M3 receptor, use of the compound of the formula (I) or a salt thereof for preventing or treating bladder/urinary tract diseases related to bladder contractions via a measuring M3 receptor, the compound of the formula (I) or a salt thereof for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M3 receptor, and a method for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M3 receptor, comprising administering to a subject an effective amount of the compound of the formula (I) or a salt thereof. Further, the “subject” is a human or a non-human animal in need of the prevention or treatment, and in one embodiment, a human in need of the prevention or treatment.
Effects of the Invention
The compound of the formula (I) or a salt thereof is a muscarinic M3 receptor-positive allosteric modulator, and can thus be used as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor.
EMBODIMENTS FOR CARRYING OUT THE INVENTION
Hereinafter, the present invention will be described in detail.
In general, the positive allosteric modulator is a compound which binds to an allosteric site different from a ligand binding site, and has an effect of increasing the affinity of an agonist to a receptor by mainly causing a structural change in a receptor, and thus changing the signal level of agonistic activity. In the living body, the positive allosteric modulator does not exhibit an agonistic effect by itself, and increases the effect of an endogenous agonist. Examples of the advantages of positive allosteric modulator over the agonists include (1) avoiding the side effects since the positive allosteric modulator exhibits an enhancement in the endogenous agonist stimulation dependently, (2) having a possibility of obtaining high subtype selectively since the positive allosteric modulator binds to a site other than a ligand binding site, and (3) less probability of causing desensitization, which can be seen with the agonists (Pharmacological Reviews, 63; pp. 59-126 (2011)).
In the present specification, the muscarinic M3 receptor-positive allosteric modulator means a compound which enhances an effect via the muscarinic M3 receptor by an agonist stimulation-dependent or nerve stimulation-dependent manner. Accordingly, only during voiding, the effect on enhancing bladder contraction is expected and the muscarinic M3 receptor-positive allosteric modulator is possibly useful as an agent for improving various symptoms associated with voiding dysfunction. Further, by such a specific action during voiding, it is expected that it is possible to avoid cholinergic side effects, known to be induced with bethanechol chloride and distigmine bromide. In addition, since the muscarinic M3 receptor-positive allosteric modulator increases bladder contractile force during voiding, an effect in voiding dysfunction which is caused by an increase in urethral resistance can also be expected. A decrease in residual urine by such improvement of voiding dysfunction leads to an increase in the effective bladder capacity, and thus, it can be expected to improve urine storage functions as well as to avoid renal disorder. Thus, the muscarinic M3 receptor-positive allosteric modulator is expected to be useful as an agent for preventing or treating bladder/urinary tract diseases related to bladder contractions via a muscarinic M3 receptor. The present inventors have newly discovered a compound that acts as the modulator, thereby completing the present invention.
In the present specification, examples of the “bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor” include voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrustor underactivity, neurogenic bladder, urethra relaxation failure, detrustor-external urethral sphincter dyssynergia, overactive bladder, urinary frequency, nocturia, urinary incontinence, benign prostatic hyperplasia, interstitial cystitis, chronic prostatitis, urethral calculus, or the like, preferably, voiding dysfunction or urine storage dysfunction in underactivity bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, and neurogenic bladder.
The “alkyl” is linear alkyl and branched alkyl. Accordingly, the “C1-6 alkyl” is linear or branched alkyl having 1 to 6 carbon atoms, and specific examples thereof include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl; in one embodiment, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, each of which is C1-4 alkyl; in one embodiment, a group selected from the group consisting of methyl, ethyl, isopropyl, and isobutyl; and in one embodiment, a group selected from the group consisting of methyl and ethyl.
The “alkylene” is linear alkylene or branched alkylene. Accordingly, the “C1-6 alkylene” is linear or branched alkylene having 1 to 6 carbon atoms, and examples thereof include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, propylene, methylmethylene, ethylethylene, 1,2-dimethylethylene, or 1,1,2,2-tetramethylethylene; in one embodiment, C1-3 alkylene; in one embodiment, methylene or ethylene; in one embodiment, methylene; and in another embodiment, ethylene.
The “halogeno-C1-6 alkyl” is C1-6 alkyl substituted with at least one halogen atom; in one embodiment, C1-6 alkyl substituted with 1 to 5 halogen atoms; in one embodiment, difluoromethyl or trifluoromethyl; and in one embodiment, trifluoromethyl.
The “cycloalkyl” is a saturated hydrocarbon cyclic group. Accordingly, the “C3-8 cycloalkyl” is a saturated hydrocarbon cyclic group having 3 to 8 ring members, and specific examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl; in one embodiment, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is C3-6 cycloalkyl; and in one embodiment, cyclopropyl.
The “aryl” is a C6-14 monocyclic to tricyclic aromatic hydrocarbon cyclic group and includes a partially hydrogenated cyclic group thereof, and specific examples thereof include phenyl, naphthyl, tetrahydronaphthyl, indanyl, or indenyl; and in one embodiment, phenyl.
The “monocyclic aromatic hetero ring” is a monocyclic aromatic hetero ring group having 5 to 7 ring members, which has 1 to 4 hetero atoms selected from the group consisting of a nitrogen atom, an oxygen atom, and a sulfur atom as a ring-constituting atom, and specific examples thereof include pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furyl, thienyl, oxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, or azepanyl; in one embodiment, thienyl or pyridyl; and in one embodiment, thienyl.
The “bicyclic aromatic hetero ring” is a bicyclic aromatic hetero ring group in which the monocyclic aromatic hetero ring is fused with a benzene ring or monocyclic aromatic hetero ring and includes a partially hydrogenated ring group thereof, and specific examples thereof include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzofuranyl, benzothienyl, benzooxazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridyl, thienopyridyl, indolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl, tetrahydroisoquinolyl, dihydrofuropyridyl, or dihydrothienopyridyl; and in one embodiment, benzothienyl.
The “saturated hetero ring” is a 3- to 8-membered saturated ring group, which has 1 to 4 hetero atoms selected from the group consisting of a nitrogen atom, an oxygen atom, and a sulfur atom as a ring-constituting atom, and may be bridged with C1-6 alkylene, in which a sulfur atom as the ring-constituting atom may be oxidized. Specific examples thereof include azepanyl, diazepanyl, oxazepanyl, thiazepanyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, piperidinyl, pyrazolidinyl, piperazinyl, azocanyl, thiomorpholinyl, thiazolindinyl, isothiazolindinyl, oxazolindinyl, morpholinyl, thiomorpholinyl, tetrahydrothiophenyl, oxathioranyl, oxiranyl, oxetanyl, dioxiranyl, tetrahydrofuranyl, tetrahydropyranyl, and 1,4-dioxanyl.
The “cyclic amino” is a 4- to 7-membered group having a bond at a ring-constituting nitrogen atom in the saturated hetero ring. Specific examples thereof include aziridin-1-yl, azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, azocan-1-yl, morpholin-4-yl, thiomorpholin-4-yl, piperazin-1-yl, 1,4-diazepan-1-yl, 1,4-oxazepan-4-yl, or 1,4-thiazepan-4-yl; in one embodiment, pyrrolidin-1-yl, piperidin-1-yl, azetidin-1-yl, morpholin-4-yl, or piperazin-1-yl, and in one embodiment, pyrrolidin-1-yl or piperidin-1-yl.
The “halogen” means fluoro, chloro, bromo, or iodo; in one embodiment, fluoro, chloro, or bromo; in one embodiment, fluoro or chloro; in one embodiment, fluoro; and in another embodiment, chloro.
In the present specification, the expression “which may be substituted” means “which is not substituted” or “which is substituted with 1 to 5 substituents”. Further, if it has a plurality of substituents, the substituents may be the same as or different from each other.
Examples of the acceptable substituent in the “cyclic amino which may be substituted”, the “C3-8 cycloalkyl which may be substituted”, the “aryl which may be substituted”, the “monocyclic aromatic hetero ring which may be substituted”, and the “bicyclic aromatic hetero ring which may be substituted” include substituents in the following Group G.
Group G
(a) C1-6 alkyl which may be substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alkyl), —CN, —SO2—(C1-6 alkyl), and halogen,
(b) —OH,
(c) —O—(C1-6 alkyl which may be substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alkyl), —CN, —SO2—(C1-6 alkyl), and halogen),
(d) C3-8 cycloalkyl,
(e) —O—(C3-8 cycloalkyl),
(f) halogen,
(g) —CN,
(h) —SO2—(C1-6 alkyl),
(i) —CO2—(C1-6 alkyl) and —COOH,
(j) —CO—N(C1-6 alkyl)2, —CO—NH(C1-6 alkyl), and —CONH2,
(k) —CO—(C1-6 alkyl),
(l) —SO2—N(C1-6 alkyl)2, —SO2—NH(C1-6 alkyl), and —SO2NH2,
(m) —N(C1-6 alkyl)2, —NH(C3-6 alkyl), and —NH2,
(n) a saturated hetero ring, and
(o) —O-saturated hetero ring.
Examples of the substituent in the “cyclic amino which may be substituted” further include oxo (═O).
In addition, the preferable substituents in the “C1-6 alkyl which may be substituted” are the substituents described in (b) to (o) of Group G above.
Examples of the preferable substituents for the “cyclic amino which may be substituted” in R1 include, in one embodiment, the substituents described in (a) to (c), (f), and (g) of Group G above; in one embodiment, C1-6 alkyl which may be substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alkyl), —CN, —SO2—(C1-6 alkyl), and halogen; in one embodiment, a group selected from the group consisting of C1-6 alkyl and halogeno-C1-6 alkyl; and in one embodiment, a group selected from the group consisting of methyl and ethyl.
Examples of the preferable substituents for the “C1-6 alkyl which may be substituted” in R12 include, in one embodiment, the substituents described in (b) to (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C3-8 cycloalkyl, —O—(C1-6 alkyl), —O—(C3-8 cycloalkyl), halogen, —CN, and cyclic amino; in one embodiment, a group selected from the group consisting of C3-8 cycloalkyl and —O—(C1-6 alkyl); and in one embodiment, a group selected from the group consisting of cyclopropyl and methoxy.
Examples of the preferable substituents for the “C3-8 cycloalkyl which may be substituted” in R13 include, in one embodiment, the substituents described in (a) to (c), (f), and (g) of Group G above; and in one embodiment, C1-6 alkyl which may be substituted with —O—(C1-6 alkyl).
Examples of the preferable substituents for the “aryl which may be substituted” in R2 include, in one embodiment, the substituents described in (a) to (d), (f), (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C1-6 alkyl, halogen-C1-6 alkyl, —O—(C1-6 alkyl), —O-(halogeno-C1-6 alkyl), halogen, C3-8 cycloalkyl, and —CN; in one embodiment, a group selected from the group consisting of halogeno-C1-6 alkyl and halogen; and in one embodiment, a group selected from the group consisting of trifluoromethyl and fluoro.
Examples of the preferable substituents for the “monocyclic aromatic hetero ring which may be substituted” and “bicyclic aromatic hetero ring which may be substituted” in R2 include, in one embodiment, the substituents described in (a) to (d), (f), (g), and (n) of Group G above; in one embodiment, a group selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), —O-(halogeno-C1-6 alkyl), halogen, C3-8 cycloalkyl, and —CN; in one embodiment, a group selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen; in one embodiment, a group selected from the group consisting of halogeno-C1-6 alkyl, —O—(C1-6 alkyl), and halogen; and in one embodiment, a group selected from the group consisting of trifluoromethyl, methoxy, and chloro.
One embodiment of the compound of the formula (I) or a salt thereof is shown below.
(1-1)
The compound of the formula (I) or a salt thereof, in which
R1 is
i. cyclic amino which may be substituted with 1 to 5 substituents selected from the group consisting of Group G and oxo, or
ii. —N(—R11)(—R12),
R11 is C1-6 alkyl, and
R12 is C1-6 alkyl which may be substituted with 1 to 5 substituents selected from the substituents described in (b) to (o) of Group G, or C3-8 cycloalkyl which may be substituted with 1 to 5 substituents selected from Group G.
(1-2)
The compound of the formula (I) or a salt thereof, in which
R1 is
i. cyclic amino which may be substituted with 1 to 5 substituents selected from the group consisting of Group G and oxo, or
ii. —N(—R11)(—R12),
R11 is C1-6 alkyl, and
R12 is C1-6 alkyl which may be substituted with 1 to 3 substituents selected from the substituents described in (b) to (g), and (n) of Group G.
(1-3)
The compounds of the formula (I) or a salt thereof, in which
R1 is
i. pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl and piperidin-1-yl are each substituted with 1 to 2 substituents selected from the group consisting of C1-6 alkyl and halogen-C1-6 alkyl, or
ii. —N(—R11)(—R12), in which
R11 is C1-6 alkyl, and
R12 is C1-6 alkyl which may be substituted with one group selected from the group consisting of C3-8 cycloalkyl and —O—(C1-6 alkyl).
(1-4)
The compound of the formula (I) or a salt thereof, in which R1 is cyclic amino substituted with 1 to 2 groups selected from the group consisting of C1-6 alkyl and halogen-C1-6 alkyl.
(1-5)
The compound of the formula (I) or a salt thereof, in which R1 is pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl and piperidin-1-yl may be substituted with 1 to 3 substituents selected from Group G.
(1-6)
The compound of the formula (I) or a salt thereof, in which R1 is pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl and piperidin-1-yl are each substituted with 1 to 2 groups selected from the group consisting of C1-6 alkyl and halogeno-C1-6 alkyl.
(1-7)
The compound of the formula (I) or a salt thereof, in which R1 is pyrrolidin-1-yl substituted with 1 to 2 groups selected from the group consisting of methyl and ethyl.
(1-8)
The compound of the formula (I) or a salt thereof, in which
R1 is —N(—R11)(—R12),
R11 is C1-6 alkyl, and
R12 is C1-6 alkyl which may be substituted with a group selected from the group consisting of C3-8 cycloalkyl and —O—(C1-6 alkyl),
(1-9)
The compound of the formula (I) or a salt thereof, in which
R1 is —N(—R11)(—R12),
R11 is methyl, ethyl, or isopropyl, and
R12 is methyl, ethyl, isopropyl, isobutyl, cyclopropylmethyl, or methoxyethyl.
(2-1)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. aryl which may be substituted with 1 to 5 substituents selected from Group G,
ii. monocyclic aromatic hetero ring which may be substituted with 1 to 5 substituents selected from Group G, or
iii. bicyclic aromatic hetero ring which may be substituted with 1 to 5 substituents selected from Group G.
(2-2)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. phenyl which may be substituted with 1 to 5 substituents selected from Group G,
ii. thienyl which may be substituted with 1 to 3 substituents selected from Group G
iii. pyridyl which may be substituted with 1 to 3 substituents selected from Group G, or
iv. benzothienyl which may be substituted with 1 to 5 substituents selected from Group G.
(2-3)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. phenyl which may be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), —O-(halogeno-C1-6 alkyl), halogen, C3-8 cycloalkyl, and —CN,
ii. thienyl which may each be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen,
iii. pyridyl which may each be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen, or
iv. benzothienyl,
(2-4)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. phenyl di-substituted with trifluoromethyl and fluoro,
ii. thienyl mono-substituted with trifluoromethyl or chloro, or
iii. pyridyl di-substituted with trifluormethyl and methoxy.
(2-5)
The compound of the formula (I) or a salt thereof, in which R2 is a monocyclic aromatic hetero ring which may be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen.
(2-6)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. thienyl which may be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen, or
ii. pyridyl which may be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen.
(2-7)
The compound of the formula (I) or a salt thereof, in which R2 is thienyl which may be substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, C3-8 cycloalkyl, and halogen.
(2-8)
The compound of the formula (I) or a salt thereof, in which R2 is thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen.
(2-9)
The compound of the formula (I) or a salt thereof, in which R2 is thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of trifluoromethyl and chloro.
(2-10)
The compound of the formula (I) or a salt thereof, in which R2 is thienyl mono-substituted with trifluoromethyl or chloro.
(2-11)
The compound of the formula (I) or a salt thereof, in which R2 is pyridyl which may be substituted with 1 to 3 groups selected from the group consisting of halogeno-C1-6 alkyl and —O—(C1-6 alkyl),
(2-12)
The compound of the formula (I) or a salt thereof, in which R2 is phenyl which may be substituted with 1 to 5 groups selected from the group consisting of C1-6 alkyl, halogen-C1-6 alkyl, —O—(C1-6 alkyl), —O-(halogeno-C1-6 alkyl), halogen, C3-8 cycloalkyl, and —CN.
(2-13)
The compound of the formula (I) or a salt thereof, in which R2 is phenyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen.
(2-14)
The compound of the formula (I) or a salt thereof, in which
R2 is
i. thienyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen, or
ii. phenyl which may be substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen.
(3-1)
The compound of the formula (I) or a salt thereof, in which R3's are the same as or different from each other, and are each C1-6 alkyl.
(3-2)
The compound of the formula (I) or a salt thereof, in which R3 is methyl.
(4-1)
The compound of the formula (I) or a salt thereof, in which W is C1-6 alkylene.
(4-2)
The compound of the formula (I) or a salt thereof, in which W is C1-3 alkylene.
(4-3)
The compound of the formula (I) or a salt thereof, in which W is methylene or ethylene.
(4-4)
The compound of the formula (I) or a salt thereof, in which W is methylene.
(4-5)
The compound of the formula (I) or a salt thereof, in which W is ethylene.
(5-1)
The compound of the formula (I) or a salt thereof, in which n is an integer of 0 to 4.
(5-2)
The compound of the formula (I) or a salt thereof, in which n is an integer of 0 to 2.
(5-3)
The compound of the formula (I) or a salt thereof, in which n is 0 or 1.
(6) The compound of the formula (I) or a salt thereof, which is a combination of any two or more of the groups, which are not inconsistent with each other, among some embodiments of each group described in (1-1) to (5-3) above. Examples thereof include the compounds or salts thereof shown below.
(6-1)
The compound of the formula (I) or a salt thereof, in which
R1 is as described in (1-2) above,
R2 is as described in (2-2) above,
R3 is as described in (3-1) above,
W is as described in (4-1) above, and
n is as described in (5-1) above.
(6-2)
The compound or a salt thereof as described in (6-1) above, in which
R1 is as described in (1-3) above,
R2 is as described in (2-3) above,
W is as described in (4-2) above, and
n is as described in (5-3) above.
(6-3)
The compound or a salt thereof as described in (6-2) above, in which
R2 is as described in (2-4) above, and
W is as described in (4-3) above.
(6-4)
The compound or a salt thereof as described in (6-2) above, in which
R1 is as described in (1-6) above,
R2 is as described in (2-14) above, and
W is as described in (4-3) above.
Examples of the specific compounds included in the present invention include the following compounds or salts thereof:
  • 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methy)pyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid,
  • 3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid,
  • [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid,
  • 3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid,
  • 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid,
  • 3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl)}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid,
  • 3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid, and
  • 3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid.
In another embodiment, examples of the specific compounds included in the present invention include the following compounds or salts thereof:
  • 3-[(3S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]propanoic acid,
  • 3-(4-{5-[(4-[6-methoxy-5-(trifluoromethyl)pyridin-3-yl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid,
  • 3-[4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoic acid,
  • [(3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]acetic acid,
  • 3-[4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoic acid,
  • 3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[isobutyl(methyl)amino]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid,
  • 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(cyclopropylmethyl)(methyl)amino]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid,
  • 3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)-thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid,
  • {(3R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-3-methylpiperazin-1-yl}acetic acid, and
  • (4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)acetic acid.
With regard to the compound of the formula (I), tautomers or geometrical isomers thereof may exist, depending on the kinds of the substituents. In the present specification, the compound of the formula (I) may be described in only one form of isomers in some cases, but the present invention includes other isomers, isolated forms of the isomers, or a mixture thereof.
Furthermore, some of the compounds of the formula (I) may have asymmetric carbon atoms or asymmetries in some cases, and correspondingly, the optical isomers thereof can exist. The present invention includes the isolated form of the optical isomer of the compound of the formula (I) or a mixture thereof.
In addition, a pharmaceutically acceptable prodrug of the compound represented by the formula (I) is also included in the present invention. The pharmaceutically acceptable prodrug refers to a compound having a group which can be converted into an amino group, a hydroxyl group, a carboxyl group, or the like, by solvolysis or under a physiological condition. Examples of the groups forming the prodrug include those as described in Prog. Med., 5, 2157-2161 (1985) or “Pharmaceutically Research and Development” (Hirokawn Publishing Company, 1990), vol. 7, Drug Design, 163-198.
Moreover, the salt of the compound of the formula (I) is a pharmaceutically acceptable salt of the compound of the formula (I), and the compounds of the formula (I) may form an acid solution salt or a salt with a base, depending on the kinds of the substituents in some cases. Specifically, examples thereof include acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid, and with organic acids such as formic acid, acetic acid, propanoic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyl tartaric acid, ditolyl tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, and glutamic acid, and salts with metal anions such as sodium, potassium, magnesium, calcium, and aluminum, and with organic bases such as methylamine, ethylamine, and ethanolamine, salts with various amino acids such as acetyl leucine, lysine, and omithine, or derivatives of amino acids, ammonium salts, and others.
In addition, the present invention also includes various hydrates or solvates, and crystal polymorph substances of the compound of the formula (I) and a salt thereof. In addition, the present invention also includes the compounds labeled with various radioactive or non-radioactive isotopes.
(Production Process)
The compound of the formula (I) or a salt thereof can be prepared by applying various know synthetic methods, using the characteristics based on their basic structures or the kinds of the substituents. At this time, depending on the types of the functional groups, it is in some cases effective from the viewpoint of the preparation techniques to protect the functional group with an appropriate protective group (a group which is capable of being easily converted into the functional groups), during the steps from starting materials to intermediates. Examples of the protective group include the protective groups as described in “Greene's Protective Groups in Organic Synthesis (4th edition, 2006)”, edited by P. G. M. Wuts and T. W. Greene, and the like, which may be appropriately selected and used depending on the reaction conditions. In these methods, a desired compound can be obtained by introducing the protective group to carry out the reaction, and then, if desired, removing the protective group.
In addition, the prodrug of the compound of the formula (I) can be prepared by introducing a specific group during the steps from starting materials to intermediates, in the same manner as for the above protective groups, or by further carrying out the reaction using the obtained compound of the formula (I). The reaction can be carried out by applying a method known to a person skilled in the art, such as common esterification, amidation, and dehydration.
Hereinbelow, typical preparation methods of the compound of the formula (I) and the compound of the formula (a) which is the starting compound will be described. Each of the production processes can also be carried out with reference to the documents appended to the description herein. Further, the preparation methods of the present invention are not limited to the examples as shown below.
(Production Process 1)
Figure USRE049111-20220621-C00006
(in which, R represents C1-6 alkyl, which shall apply hereinafter).
This reaction is a method for producing a compound of the formula (I) which is a compound of the present invention, by deprotecting a compound of the formula (a).
This reaction is carried out using the compound of the formula (a) and a deprotecting reagent in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to heating to reflux, usually for 0.1 hours to 5 days, in a solvent which is insert to the reaction or without a solvent. Examples of the solvent used herein are not particularly limited, but include alcohols such as methanol, ethanol, n-propanol and the like, N,N-dimethylformamide, tetrahydrofuran, and the like. Further, there are some cases where a mixed solvent of the solvent and water is highly suitable for the reaction. Examples of the deprotecting reagent are not particularly limited, but include bases such as an aqueous sodium hydroxide solution, an aqueous potassium hydroxide solution and the like, and acids such as hydrochloric acid, trifluoroacetic acid and the like.
(Production Process 2)
Figure USRE049111-20220621-C00007
(in which, L1 represents a leaving group, which shall apply hereinafter).
This production process is a method for producing the compound of the formula (a) which is a starting material of the compound of the formula (I). Here, examples of L1 include chloro and the like.
(Step 1)
This step is a step of preparing a compound of the formula (d) by subjecting a compound of the formula (b) and a compound of the formula (c) to an amidation reaction.
The reaction is carried out using the formula (b) and the compound of the formula (c) in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to under heating, preferably at −20° C. to 60° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction, in the presence of a condensing agent. Examples of the solvent used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane, cyclopentylmethyl ether and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, water, and a mixture thereof. Examples of the condensing reagent include 1-(3-dimethylamino propyl)-3-ethylcarbodiimide or a hydrochloride thereof, dicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole, diphenylphosphoric azide, phosphorous oxychloride, N-[({[(1Z)-1-cyano-2-ethoxy-2-oxoethylidene]amino}oxy)morpholin-4-yl)methylene]-N-methylmethanaminium hexafluorophosphate (COMU), and the like, but are not limited thereto. It may be preferable in some cases for the reaction to use an additive (for example, 1-hydroxybenzotriazole), and it may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like, or an inorganic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
Furthermore, a method in which the carboxylic acid (c) is converted to a reactive derivative thereof, and then the reactive derivative is reacted with the amine (b) can also be used. Examples of the reactive derivative of the carboxylic acid include acid halides obtained by the reaction with a halogenating agent such as phosphorus oxychloride, thionyl chloride or the like, mixed acid anhydrides obtained by the reaction with isobutyl chloroformate or the like, and active esters obtained by condensation with 1-hydroxybenzotriazole or the like. The reaction of these reactive derivatives and the compound (b) can be carried out under the temperature condition ranging from under cooling to under heating, preferably at −20° C. to 60° C., in a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers and the like.
References
  • “Organic Functional Group Preparations” written by S. R. Sandler and W. Karo, 2nd edition, Vol. 1, Academic Press Inc., 1991
  • “Courses in Experimental Chemistry (5th edition)” edited by The Chemical Society of Japan, Vol. 16 (2005) (Maruzen).
(Step 2)
This step is a step of preparing a compound of the formula (f) by reacting a compound of the formula (d) with a compound of the formula (e).
This reaction is carried out using the formula (d) and the compound of the formula (e) in equivalent amounts, or either thereof in an excess amount, by stirring the mixture under the temperature condition ranging from under cooling to under heating to reflux, preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like, or an organic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
References
  • “Organic Functional Group Preparations” written by S. R. Sandler and W. Karo, 2nd edition, Vol. 1, Academic Press Inc., 1991
  • “Courses in Experimental Chemistry (5th edition)” edited by The Chemical Society of Japan, Vol. 14 (2005) (Marazen).
(Step 3)
This step is a step of preparing a compound of the formula (g) by introducing an acetoxymethyl group into the 5-position of thiazole in the compound of the formula (f). The compound of the formula (f) is reacted with an aqueous formaldehyde solution or paraformaldehyde in the presence of an acetic acid solvent, which can be carried out under the temperature condition ranging from at room temperature to under heating to reflux. Further, the reaction can also be carried out by adding acetic acid into a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers and the like, instead of the acetic acid solvent. In addition, the reaction can also be carried out by further adding acetic anhydride.
(Step 4)
This step is a step of preparing a compound of the formula (a) by reacting a compound of the formula (g) with a compound of the formula (h) under a basic condition. The present reaction can be carried out by reacting the compound of the formula (g) with the compound of the formula (h) in the presence of an organic base such as triethylamine and N,N-diisopropylethylamine, in an organic solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers, esters, acetonitrile, N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone and the like. Further, the compound of the formula (h) may also used in an excess amount instead of the organic base. The reaction can be carried out under the temperature condition ranging from under cooling to at room temperature; from at room temperature to under heating; or from at room temperature to under refluxing.
In addition, the compound of the formula (a) can be directly obtained while not isolating the compound of the formula (g) by adding the compound of the formula (h) into the reaction mixture of Step 3.
(Production Process 3)
Figure USRE049111-20220621-C00008
(in which P1 and P2 each represent a protective group, and L2 represents a leaving group).
This production process is another preparation method for the compound of the formula (a), which is a starting material of the compound of the formula (I). Here, as the protective groups represented by P1 and P2, the groups of amino groups described in “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4th edition, John Wiley & Sons Inc., 2006, and the like can be used. Examples of the P1 include acetyl, trifluoroacetyl and the like, examples of P2 include t-butoxycarbonyl and the like, and examples of L2 include bromo and the like.
(Step 2)
This compound is a step of protecting the amino group of the compound (b). Here, the present reaction can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4th edition, John Wiley & Sons Inc., 2006.
(Step 2)
This step is a step of preparing a compound of the formula (k) by introducing an acetoxymethyl group into the 5-position of thiazole in a compound of the formula (j). The reaction conditions are the same as in Step 3 of Production Process 2.
(Step 3)
The step is a step of preparing a compound of the formula (m) by reacting a compound of the formula (h) and a compound of the formula (k) under a basic condition. The reaction conditions are the same as in Step 4 of Production Process 2.
(Step 4)
This step is a step of deprotecting a protective group P1 of an amino group of the compound (m). Here, the present reaction can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4th edition, John Wiley & Sons Inc., 2006.
(Step 5)
This step is a step of obtaining a compound of the formula (q) by subjecting a compound of the formula (o) and a compound of the formula (p) to an amidation reaction. The reaction conditions are the same as in Step 1 of Production Process 2.
(Step 6)
This step is a step of preparing a compound of the formula (s) by reacting a compound of the formula (q) with a compound of the formula (r). The reaction conditions are the same as in Step 2 of Production Process 2.
(Step 7)
This step is a step of deprotecting a protective group P2 of a compound of the formula (s).
This step can be carried out with reference to “Protective Groups in Organic Synthesis” written by Wuts and Greene, 4th edition, John Wiley & Sons Inc., 2006”.
(Step 8)
This step is a step of obtaining the compound of the formula (a) by reacting a compound of the formula (t) and a compound of the formula (n). The present reaction is carried out using the compound (t) and the compound (u) in equivalent amounts, or either thereof in an excess amount, and stirring the mixture under the temperature condition ranging from under cooling to under heating to reflux, preferably at 0° C. to 100° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction, or without a solvent. Examples of the solvent used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform and the like, N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine and the like, or an inorganic base such as potassium carbonate, sodium carbonate, potassium hydroxide and the like.
References
  • “Organic Functional Group Preparations” written by S. R. Sandler and W. Karo, 2nd edition, Vol. 1, Academic Press Inc., 1991
  • “Courses in Experimental Chemistry (5th edition)” edited by The Chemical Society of Japan, Vol. 14 (2005) (Maruzen).
The compound of the formula (I) is isolated and purified as its free compound, or a salt, a hydrate, a solvate, or crystal polymorph substance thereof. The salt of the compound of the formula (I) can also be prepared by a conventional method.
Isolation and purification are carried out by employing general chemical operations such as extraction, fractional crystallization, and various types of fractional chromatography.
Various isomers can be prepared by selecting appropriate starting compound, or separated by separation using differences in the physicochemical properties among the isomers. For example, the optical isomers can be obtained by means of general optical resolution methods of racemic compounds (for example, fractional crystallization introducing the compound into a diastereomer salt with an optically active base or acid; chromatography using a chiral column or the like; and others), or can also be prepared from appropriate optically active starting compound.
The pharmacological activity of the compound of the formula (I) was confirmed by the following test.
Test Example 1: Evaluation of Muscarinic M3 Receptor Positive Allosteric Modulator Activity
a) Construction of Vector Expressing Human Muscarinic M3 Receptor
A human muscarinic M3 receptor gene (GenBank Accession No.: NM_00740.2) was introduced into an expression vector pcDNA3.1™ (Life Technologies).
b) Construction of Cells Stably Expressing Human Muscarinic M3 Receptor
A vector expressing a human muscarinic M3 receptor was introduced into a CHO—K1 cell (ATCC No.: CCL-61). The introduction was carried out according to the attached instructions, using a transfection reagent, Lipofectoamine (registered trademark) 2000 Reagent (Life Technologies). The cells were incubated in an alpha Modified Eagle Minimum Essential Medium (α-MEM) including 2 mM glutamine, 10% fetal bovine serum, and 2.0 mg/mL. Geneticin (registered trademark) (Life Technologies) for 4 weeks to acquire a drug-resistant clone.
c) Measurement of Intracellular Ca2+ Concentration
The cells obtained in b) above were suspended in an α-MEM including 2 mM glutamine, 10% fetal bovine serum, and 0.2 mg/mL. Geneticin (registered trademark) to the amount from 1.2 to 1.5×104 cells/well the day before the experiment, dispersed into a 384-well plate (Model No. 355962, BD Biosciences), and incubated overnight at 37° C. and 5% CO2. The medium was replaced with a loading buffer (an assay buffer (Hank's balanced salt solution (HBSS), 1 g/L BSA, 20 mM HEPES (pH 7.5), and 2.5 mM probenecid), including 3.1 μM Fluo 4-AM (Dojindo Laboratories) and incubated for about 2 hours at room temperature. Thereafter, the cells were washed with a plate washer EL×405™ (BIO-TEK Instrument, Inc.) set with the assay buffer, and set in an intracellular Ca2+ concentration measuring system (FLIPRintra (registered trademark), Molecular Device Co.). The test substances (final concentration of 1 μM or 10 μM) and carbachol (Sigma, final concentration of 0.0024 nM to 10 μM) which had each been dissolved in the assay buffer in advance were set in a FLIPRintra (registered trademark). The test substances were added to the cells in the device and after about 5 minutes, carbachol was added to the cells. An increase rate of the intracellular Ca2+ concentration by carbachol was measured (excitement wavelength of 470 nm to 495 nm and a fluorescence wavelength of 515 nm to 575 nm).
For the muscarinic M3 receptor-positive allosteric modulator activity, a shift toward a lower concentration side of a carbachol concentration-response curve by the test substance was used as an index. That is, a minimum value in the carbachol response was taken as 0%; a minimum value in the carbachol response was taken as 100% from the concentration-response curve of carbachol; the carbachol concentration exhibiting a 50% response was calculated as an EC50 value, using a Sigmoid-Emax model non-linear regression method, and thus, the muscarinic M3 receptor-positive allosteric modulator activity was determined by dividing the EC50 value of carbachol in the absence of the test substance by the EC50 value of carbachol in the presence of the test substance. For example, when the EC50 value of the carbachol in the absence of the test substance was 0.1 μM and the EC50 value of carbachol in the presence of the test substance was 0.01 μM, the value of the muscarinic M3 receptor-positive allosteric modulator activity becomes 10, showing that the test substance causes a 10-fold shift in the EC50 value toward to the low concentration side. In Tables below, the columns of 10 μM (-fold shift) show the values in a case where the test substance is added to a final concentration of 10 μM and the columns of 1 μM (-fold shift) show the values in a case where the test substance is added to a final concentration of 1 μM.
Test Example 2: Evaluation of Human c-Mpl-Introduced Ba/F3 Cell Proliferation Activity
The human c-Mpl-introduced Ba/F3 cell proliferation action was measured by the following method.
As a positive control, 1-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}-3-fluoropyridin-2-yl)piperidine-4 -carboxylic acid hydrochloride disclosed as Example 315 in Patent Document 1, represented by the formula (A1) above, was used. Further, it is known that the compound has a good human c-Mpl-introduced Ba/F3 cell proliferative activity as disclosed in Table 1 in Patent Document 1.
a) Construction of Vector Expressing Human c-Mpl Receptor
A human c-Mpl receptor gene (GenBank Accession No.: M90102.1) was transfected into an expression vector pEF-BOS (Nucleic Acids Research, 18; pp 4322 (1990)).
b) Concentration of Cell Stably Expressing Human c-Mpl Receptor
A vector expressing a human c-Mpl receptor was introduced into a Ba/F3 cell (RIKEN BRC: RCB0805). For the introduction, an electroporation method was used. pEF-BOS-c-mpl (10 μg), pSV2bsr (1 μg, Kaken Pharmaceutical Co., Ltd.) and 1×109 of Ba/F3 cells were put into cuvettes with a gap width of 0.4 cm and electroporated under a condition of 1.5 kV (25 μF) in a Gene Pulser (registered trademark) (BioRad). The cells were incubated in an RPMI-1640 medium supplemented with a 0.5% WEHI conditioned medium (BD Biosciences) and 10% fetal bovine serum for 3 days, and thereafter, and the cells were incubated for 30 days in an RPMI-1640 medium, to which 10 μg/mL blasticidin had been further added, thereby acquiring a drug-resistance clone.
c) Measurement of Cell Proliferative Activity
The cells obtained in b) above were dispersed into an RPMI-1640 medium supplemented with a 0.5% WEHI conditioned medium and 10% fetal bovine serum, and used. The day before the experiment, the test substances (final concentration of 100 mM to 10 μM) which has been dissolved in a medium for assay (an RPMI-1640 medium supplemented with 10% fetal bovine serum) were added to a 384-well plate (Model No. 781185, Greiner bio-one). The cells after the medium had been replaced with the medium for assay were dispensed to a 384-well plate to which the test substance had been added, to 1×104 cells/well, and incubated overnight at 37° C. and 5% CO2. On the experiment day, a solution of a Cell counting kit (Dojindo Laboratories) was added to each well of the 384-well plate, and the cells were incubated for about 5 hours at 37° C., and 5% CO2. Thereafter, the absorbance (an absorbance wavelength of 450 nm) of each well was measured using Safire2 (registered trademark) (TECAN), and used as an index for the number of cells. Further, as a negative control, a well to which the test substances had not been added was prepared.
By taking the absorbance of the well to which the test substance had been not added as 0% and taking the absorbance in a case where the positive control had been added to a final concentration of 1 μM as 100%, a cell proliferation rate (%) was calculated from the absorbance of the well to which the test substance has been added. From the obtained results, the test substance concentration exhibiting 30% proliferation by a Sigmoid-Emax model non-linear regression method was calculated as an EC30 value.
Combinations of the muscarinic M3 receptor-positive allosteric modulator activity (-fold shift) and the human c-Mpl-introduced Ba/F3 cell proliferative activity (EC30 value, nM) of some Example compounds of the present invention are shown in Tables 1 and 2. However, Ex represents Example Compound Nos. as described later (this shall apply hereinafter).
TABLE 1
Test Example 1 Test Example 2
10 μM 1 μM EC30
Ex (-fold shift) (-fold shift) (nM)
 3 253 101 780
 4 200  25 >3000
10  87  21 >10000
11 226  33 >10000
12 178  33 >10000
13 326  43 >10000
15 159  31 >10000
17 109  15 >10000
21 149  25 >10000
27 330  31 >10000
28 108  36 5300
33 182  40 >10000
34 116  18 >10000
41 160  43 >10000
42 141  39 >10000
43 224  76 >10000
46 199  29 >10000
48 113  27 >10000
49 224  67 >10000
50 190 108 2300
51 287 102 2600
52 196  36 >10000
54 134  36 >10000
60 235  33 9700
61 229  35 1300
62 195  37 >10000
63 186  39 >10000
64 128  23 >10000
65  90  24 >10000
67 114  40 >10000
69 177  27 >10000
TABLE 2
Test Example 1 Test Example 2
10 μM 1 μM EC30
Ex (-fold shift) (-fold shift) (nM)
 71 151  28 >10000
 72 152  31 >10000
 79 171  60 >10000
 81  94  89 1800
 82  43  11 500
 91 139  19 >10000
 92 203  30 >10000
 95 233  91 >10000
 97 121  55 3000
100 229  82 2800
101 112  64 3200
103 307 202 2700
104 195  75 1700
106 270  41 >10000
107 318  73 >10000
108 169  56 >10000
109 191  30 >10000
111 627 203 5000
118 167  57 >10000
119 503 110 >10000
124 101  28 >10000
126 318  79 >10000
128 192  73 8000
129 148  67 >10000
130 151  95 >10000
132  41  15 >10000
133 164  30 >10000
135 204  25 >10000
140 158  28 >10000
141 159  45 >10000
142 160  52 4700
143  81  65 7800
In Test Example 1, a substantial number of the Example compounds which had been subjected to the present test shifted the EC50 values to almost 100-fold or more toward a lower concentration side when added at 10 μM, and shifted the EC50 values to almost 10-fold or more toward a lower concentration side when added at 1 μM. In addition, for some Example compounds of the present invention, from the viewpoint that the compounds alone do not change the intracellular Ca2+ concentration, it was found that these compounds have no muscarinic M3 receptor agonistic activity.
Furthermore, in Test Example 2, it was found that a substantial number of the Example compounds which had been subjected to the present test have a weak human c-Mpl-introduced Ba/F3 cell proliferative activity or have none.
The compound of the present invention is used as an agent for preventing or treating bladder/urinary tract diseases associated with bladder concentrations via a muscarinic M3 receptor, as a muscarinic M3 receptor-positive allosteric modulator, and thus preferably has a weak or none increased platelet action based on c-Mpl-introduced Ba/F3 cell proliferative activity.
On the other hand, Table 1 of Patent Document 1 above discloses that the compound of Example 315 represented by the formula (A1) above has 3.2 nM of EC30 value of c-Mpl-introduced Ba/F3 cell proliferation action.
Test Example 3: Effect on Electrical Field Stimulation-Induced Contraction in Rat Isolated Bladder
As an effect on the nerve stimulation-dependent bladder contraction in in vitro, the effect of the Example compounds of the present invention in the electrical field stimulation-induced contraction of the rat isolated bladder was measured by the following method. That is, a bladder specimen having a width of about 2 mm and a length of about 10 mm in the longitudinal direction from the bladder isolated from a Sprague-Dawley (SD) female rat (Japan SLC, Inc.) was prepared. The prepared bladder specimen was suspended in an organ bath filled with 10 mL of a Krebs-Henseleite solution. The Krebs-Henseleite solution was aerated at 95% 02 and 5% CO2, and kept at 37° C. After carrying out stabilization at an initial tension of 1 g, the contraction was caused twice with 60 nM KCl. After stabilization of the specimen with a Krebs-Henseleite solution, the concentration was caused by carrying out electrical field stimulation at 20 V with an electrical stimulation device (Nihon Kohden) (a stimulation frequency of 8 Hz, a pulse width of 0.3 msec, and a stimulation time of 10 seconds). By repeating the transmural electrical stimulation at an interval of 2 minutes, a voltage was adjusted to obtain a contraction height of approximately 50% of the contractile response at 20 V. After the contraction by electrical field stimulation had been stabilized, 10 μL of the test substances dissolved in 100% dimethyl sulfoxide in advance (final concentrations of 3 μM, 10 μM, and 30 μM) was added thereto. The test substances were cumulatively administered at the following concentrations after the low-concentration contractile response had been stabilized. The response was taken into a personal computer through a PowerLab (registered trademark) (AD Instruments, Inc.), and analyzed by LabChart (registered trademark) (AD Instruments, Inc.). When the area under the response (area under curve, AUC) in each contraction response was calculated and the value before treatment with the test substance was taken as 100%, the enhancement rate (% of pre) of the isolated bladder concentrations after treatment with the test substance was calculated.
The enhancement rates of the isolated bladder contractions by 10 μM of some Example compounds are shown in Table 3.
Furthermore, it was confirmed that all the Example compounds which have been subjected to the present test do not cause contraction in a state in which there is no electrical stimulation and the compounds alone do not show a bladder contraction action.
TABLE 3
Enhancement rate (% of pre) of
Ex. isolated bladder contractions
  3 152
 10 161
 11 123
 13 126
 15 124
 21 141
 28 123
 34 137
 42 158
 43 179
 46 132
 48 143
 49 153
 50 183
 51 151
 52 132
 60 144
 61 176
 64 162
 65 127
 67 116
 72 157
 82 158
 95 150
109 183
119 154
124 132
133 151
135 139
140 161
141 121
142 196
143 140
From the above, it was confirmed that the Example compounds alone, which have been subjected to the present test, do not cause a contraction action in the isolated rate bladder, but have an action of enhancing electrical field stimulation-induced contraction.
Test Example 4: Effect on Pelvic Nerve Stimulation-Induced Elevation of Intravesical Pressure in Anesthetized Rats
The effect of the Example compounds of the present invention in the pelvic nerve electrical stimulation-induced elevation of intravesical pressure using rats as an action of nerve stimulation-dependent bladder contraction in vivo was measured by the following method. That is, SD female rats (Japan SLC, Inc.) were used and its lower abdomen was dissected at the midline under pentobarbital anesthesia (50 mg/kg ip). After ligating and cutting the ureter on both sides, a cannula (PE-5) for measuring the intravesical pressure was inserted into the bladder from the external urethral opening and fixed b a clip. After injecting about 200 μL of saline through the cannula that had been inserted into the bladder, the other side was connected to a pressure transducer to measure the intravesical pressure. Under a stereoscopic microscope observation, the pelvic nerve in the vicinity of the bladder was peeled and an electrode for nerve stimulation (unique Medical) was placed. The abdominal cavity was filled with mineral oil (MP BIOMEDICALS). After placing in a post-operative stabilization period, the pelvic nerve was subjected to electrical stimulation (stimulation voltage: 10 V, stimulation frequency: 8 Hz, pulse width: 0.3 msec, and stimulation time: 10 seconds) to elicit the elevation of intravesical pressure, using an electrical stimulator (Nihon Kohden). By repeating the electrical stimulation at an interval of 2 minutes while adjusting the voltage, the voltage was adjusted to elicit about 50% to 70% elevation of intravesical pressure elicited at 10 V. Thereafter, by repeating the electrical stimulation at an interval of 10 minutes, the increase in the intravesical pressure by electrical stimulation was stabilized three times or more, and the test substance (an administration amount of 3 mg/kg) was then administered from the catheter detained in the vein at a volume of 1 mL/kg, thus measuring an effect of the elevation of the intravesical pressure of the test substance for 1 hour. The test substance was dissolved in water supplemented with 10% dimethylsulfoxide and 10% Cremophor.
The response was applied to a personal computer through a PowerLab (registered trademark) and analyzed by Lab-Chart (registered trademark). The AUC of each elevation of the intravesical pressure was calculated, the intravesical pressure elevation rate (% of pre) after the treatment with the test substance was calculated by taking an average value of the values measured three times before the treatment with the test substance as 100%, and the maximum effect during a period within one hour after administration of the compound was considered as the effect of the test substance.
The elevation rates (% of pre) of the intravesical pressure when some Example compounds were administered at 3 mg/kg are shown in Table 4.
TABLE 4
Enhancement rate (% of pre) of
Ex. isolated bladder contractions
  3 251
 10 145
 11 132
 13 132
 15 142
 21 155
 28 184
 34 134
 42 149
 43 125
 46 126
 48 121
 49 172
 50 207
 51 223
 52 129
 60 130
 61 129
 64 135
 65 128
 67 126
 72 155
 82 138
 95 239
109 180
119 173
124 143
133 150
135 168
140 148
141 175
142 199
143 172
In addition, it was confirmed that the Example compounds evaluated in the present test do not cause an elevation of the intravesical pressure in a state in which electrical stimulation is not given, and the compounds alone do not show elevation of the intravesical pressure.
From the above, it was confirmed that the Example compounds listed in Table 4 alone do not show elevation of the intravesical pressure but have an action of enhancing effect on the pelvic nerve electrical stimulation-induced elevation of intravesical pressure in the anesthetized rats.
As shown in the results of each of the tests above, it was confirmed that the compound of the formula (I) has a muscarinic M3 receptor-positive allosteric modulator activity, and further, it enhances the bladder contraction in a nerve stimulation-dependent manner in in vitro, as well as enhances an elevation in the intravesical pressure in a nerve stimulation-dependent manner in in vitro. Accordingly, the compound of the formula (I) can be used to prevent or treat bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, in particular, voiding dysfunction or urine storage dysfunction in the bladder/urethral diseases. The compound of the formula (I) can be used for preventing or treating, for example, voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, neurogenic bladder, urethra relaxation failure, detrusor-external urethral sphincter dyssynergia, overactive bladder, urinary frequency, nocturia, urinary incontinence, benign prostatic hyperplasia, interstitial cystitis, chronic prostatitis, and urinary tract stones. In particular, the compound of the formula (I) can be used for preventing or treating voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, and neurogenic bladder.
In addition, the compound of formula (I) can become a therapeutic drug that is more excellent in safety from the viewpoint that the compound alone does not show an agonistic effect on a muscarinic M3 receptor, but shows an effect on enhancing the nerve stimulation-dependent bladder contraction, and accordingly, cholinergic side effects that have been reported in the existing drugs can be avoided.
A pharmaceutical composition including one or two or more kinds of the compound of the formula (I) as an active ingredient can be prepared using an excipient which is usually used in the art, that is, an excipient for a pharmaceutical preparation, a carrier for a pharmaceutical preparation, and the like, according to a method usually used.
Administration can be accomplished either by oral administration via tablets, pills, capsules, granules, powders, solutions, and the like, or parenteral administration via injections, such as intraarticular, intravenous, and intramuscular injections, suppositories, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalers, and the like.
As a solid composition for oral administration, tablets, powders, granules, and the like are used. In such a solid composition, one kind or two or more kinds of the active ingredients are mixed with at least one inactive excipient. In a conventional method, the composition may contain inactive additives such as a lubricant, a disintegrating agent, a stabilizer, or a solubilization assisting agent. If necessary, tablets or pills may be coated with a sugar or with a film of a gastric or enteric coating substance.
The liquid composition for oral administration includes pharmaceutically acceptable emulsions, solutions, suspensions, syrups elixirs, or the like, and also includes generally used inert diluents, for example, purified water or ethanol. The liquid composition may also include auxiliary agents such as a solubilization assisting agent, a moisturizing agent, and a suspending agent, sweeteners, flavors, aromatics, and antiseptics, in addition to the inert diluent.
The injections for parenteral administration include sterile aqueous or non-aqueous solution preparations, suspensions, or emulsions. The aqueous solvent includes, for example, distilled water for injection and saline. Examples of the non-aqueous solvent include alcohols such as ethanol. Such a composition may further include a tonicity agent, an antiseptic, a moistening agent, an emulsifying agent, a dispersing agent, a stabilizing agent, or a solubilizing assisting agent. These are stabilized, for example, by filtration through a bacteria retaining filter, blending of a bactericide, or irradiation. In addition, these can also be used by preparing a sterile solid composition, and dissolving or suspending it in sterile water or a sterile solvent for injection prior to its use.
Examples of the agent for external use include ointments, hard plasters, creams, jellies, cataplasms, sprays, and lotions. The agent further contains generally used ointment bases, lotion bases, aqueous or non-aqueous liquid preparations, suspensions, emulsions, or the like.
As the transmucosal agents such as an inhaler and a transnasal agent, those in the form of a solid, liquid, or semi-solid state are used, and can be prepared in accordance with a method known in the related art. For example, a known excipient, and also a pH adjusting agent, an antiseptic, a surfactant, a lubricant, a stabilizing agent, a thickening agent, or the like may be appropriately added thereto. For the administration, an appropriate device for inhalation or blowing can be used. For example, a compound may be administered alone or as a powder of formulated mixture, or as a solution or suspension in combination with a pharmaceutically acceptable carrier, using a known device or sprayer such as a metered administration inhalation device. A dry powder inhaler or the like may be for single or multiple administration use, and a dry powder or a powder-containing capsule may be used. Alternatively, this may be in a form such as a pressurized aerosol spray that uses an appropriate propellant agent, for example, a suitable gas such as chlorofluoroalkanes, and carbon dioxide, or other forms.
Usually, in the case of oral administration, the daily dose is from about 0.001 mg/kg to 100 mg/kg, preferably from 0.1 mg/kg to 30 mg/kg, and more preferably from 0.1 mg/kg to 10 mg/kg, per body weight, administered in one portion or in 2 to 4 divided portions. In the case of intravenous administration, the daily dose is suitably administered from about 0.0001 mg/kg to 10 mg/kg per body weight, once a day or two or more times a day. In addition, a transmucosal agent is administered at a dose from about 0.001 mg/kg to 100 mg/kg per body weight, once or plural times a day. The dose is appropriately decided in response to the individual case by taking the symptoms, the age, and the gender, and the like into consideration.
Although there are differences depending on a route of administration, a dosage form, an administration site, and a type of the excipient or additive, a pharmaceutical composition of the present invention comprises 0.001% by weight to 100% by weight of, as an embodiment, 0.01% by weight to 50% by weight of, one or more of the compound of the formula (I) or a salt thereof which is the active ingredient.
The compound of the formula (I) may be used in combination with various agents for treating or preventing diseases on which the compound of the formula (I) is considered to show the effect. Such combined preparations may be administered simultaneously, or separately and continuously, or at a desired time interval. The preparations to be co-administered may be a blend, or may be prepared individually.
EXAMPLES
Hereinbelow, the production process for the compound of the formula (I) will be described in more detail with reference to Examples. Further, the present invention is not limited to the compounds described in the Examples below. Further, the production processes for the starting compounds will be described in Preparation Examples. In addition, the production processes for the compound of the formula (I) are not limited to the production processes of the specific Examples shown below, but the compound of the formula (I) can be prepared by a combination of these production processes or a method that is apparent to a person skilled in the art.
Further, in the present specification, nomenclature software such as ACDC/Name (registered trademark, Advanced Chemistry Development, Inc.) may be used for nomenclature of compounds in some cases.
The powder X-ray diffraction is measured using RINT-TTRII under the condition of a tube: Cu, a tube current: 300 mA, a tube voltage: 50 kV, a sampling width: 0.020°, a scanning speed: 4°/min, a wavelength: 1.54056 angstroms, and a measurement diffraction angle (2θ): 2.5° to 40°. Further, a device including data processing was handled in accordance with the method and procedure instructed in each device.
The values obtained from various spectra may cause some errors according to the direction of the crystal growth, particle sizes, measurement conditions, and the like in some cases. Accordingly, considering these errors, in the present specification, the description of diffraction angles (2θ (°)) in the powder X-ray diffraction patterns is measured value, but depending on the measuring conditions, these diffraction angles mean that error ranges which are usually acceptable may occur and means that they are approximate values. Usually, the error range of the diffraction angle (2θ (°)) in the powder X-ray diffraction is ±0.2°. However, for the powder X-ray diffraction patterns, in terms of the properties of data, crystal lattice spacing and general patterns are important in the certification of crystal identity, and the diffraction angle and the diffraction intensity may vary slightly depending on the direction of crystal growth, the particle size, and the measurement condition, and they should not be strictly construed.
Moreover, the following abbreviations may be used in Examples, Preparation Examples, and Tables below in some cases.
PEx: Preparation Example No., Ex: Example No., PSyn: Preparation Example No. prepared by the same method, Syn: Example No. prepared by the same method, Structure: Structural chemical formula (Me represents methyl, Et represents ethyl, Ac represents acetyl, nPr represnets n-propyl, iPr represents isopropyl, cPr represents cyclopropyl, iBu represents isobutyl, Boc represents tert-butoxycarbonyl, Ts represents 4-methylphenyl sulfonyl, COMU represents N-[({[(1Z)-1-cyano-2-ethoxy-2-oxoethylidene]amino}oxy) (morpholin-4-yl)methylene]-N-methylmethaminium hexafluorophosphate, WSCD.HCl represents N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimide monohydrochloride, and ODS represents octadecylsilyl), Data: Physicochemical data, ESI+: m/z values in mass spectroscopy (Ionization method ESI, representing [M+H]+ unless otherwise specified), ESI−: m/z values in mass spectroscopy (Ionization method ESI, representing [M−H]− unless otherwise specified), APCI/ESI+: APCI/ESI-MS (atmospheric pressure chemical ionization method APCI, representing [M+H]+ unless otherwise specified; in which APCI/ESI means simultaneous measurement of APCI and ESI), EI: m/z values in mass spectroscopy (Ionization method EI, representing [M]+ unless otherwise specified), CI: m/z values in mass spectroscopy (Ionization method CI, representing [M+H]+ unless otherwise specified), NMR-DMSO-d: δ (ppm) of peaks in 1H-NMR in DMSO-d6, s: singlet (spectrum), d: doublet (spectrum), t: triplet (spectrum), br: broad line (spectrum) (e.g.: brs), m: multiplet (spectrum). Further, HCl in the structural formula indicates that the compound is a monohydrochloride; 2HCl indicates that the compound is a dihydrochloride: 3HCl indicates that the compound is a trihydrochloride, and 2 maleic acid indicates that the compound is a dimalate dimaleate.
In addition, for the sake of convenience, a concentration of mol/L is represented by M. For example, a 1 M aqueous sodium hydroxide solution means a 1 mol/L aqueous sodium hydroxide solution.
Preparation Example 1
A mixture of 4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazolyl-2-amine (1.0 g), 5-chloropyrazine-2-carboxylic acid (685 mg), COMU (1.9 g), dioxane (10 mL), and N,N-diisopropylethylamine (1.5 mL) was stirred at room temperature for 1 hour. The reaction mixture was diluted with ethyl acetate, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 5-chloro-N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{([(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)pyrazine-2-carboxamide (800 mg) as a solid.
Preparation Example 2
To a mixture of 5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-amine (2.9 g) and dichloromethane (60 mL) were added 5-chloropyrazine-2-carboxylic acid (1.7 g), N,N-dimethyl-4-aminopyridine (340 mg), and WSCD.HCl (2.1 g), followed by stirring at 40° C. for 15 minutes. The reaction mixture was cooled to room temperature, diluted with chloroform, and washed with a saturated aqueous sodium hydrogen carbonate solution. The aqueous layer was extracted with chloroform/methanol and the organic layer was combined and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-ethyl acetate) to obtain 5-chloro-N-(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide (2.4 g) as a solid.
Preparation Example 3
To a mixture of 5-chloropyrazine-2-carboxylic acid (30.5 g) and ethyl acetate (500 mL) were added thionyl chloride (55 mL) and N,N-dimethylformamide (0.57 mL), followed by stirring at 75° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure and toluene was added thereto, followed by carrying out a concentration operation.
A mixture of 4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-amine (3.20 g) and cyclopentylmethyl ether (500 mL) was ice-cooled, and triethylamine (62 mL), and a mixture of the previously obtained compound and cyclopentyl ether (100 mL) were slowly added thereto. The reaction mixture was stirred at room temperature for 2-days. To the reaction mixture was added water, followed by extraction with ethyl acetate/tetrahydrofuran. The organic layer was washed with water and saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was mixed with diisopropyl ether and the solid was collected by filtration to obtain 5-chloro-N-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]pyrazine-2-carboxamide (46.6 g) as a solid.
Preparation Example 4
To a mixture of 6-methoxy-5-(trifluoromethyl)nicotinic acid (7.8 g) and dichloromethane (80 mL) were added N,O-dimethylhydroxylamine hydrochloride (4.3 g), WCSCD.HCl (9.5 g), and N,N-diisopropylethylamine (30 mL) under ice-cooling. The reaction mixture was stirred at room temperature for 17 hours. The reaction mixture was concentrated under reduced pressure, and to the residue were added ethyl acetate and water, followed by stirring for 30 minutes. The organic layer was separated, the aqueous layer was extracted with ethyl acetate, and the organic layer was combined, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain N,6-dimethoxy-N-methyl-5-(trifluoromethyl)nicotinamide (5.0 g) as an oil.
Preparation Example 5
A mixture of N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl) acetamide (1.4 g), ethanol (10 mL), and a 6 M aqueous sodium hydroxide solution (5 mL) was stirred at 120° C. for 15 minutes under microwave irradiation. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-amine (1.0 g) as an oil.
Preparation Example 6
A mixture of N-(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)acetamide (916 mg) and 80% sulfuric acid (10 mL) was stirred at 100° C. for 1 hour. The reaction mixture was cooled to 5° C. and alkalified by the addition of a 5 M aqueous sodium hydroxide solution and a saturated aqueous sodium hydrogen carbonate solution. The mixture was extracted with chloroform, and the organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain 5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (685 mg) as a solid.
Preparation Example 7
To a mixture of N-{5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide (392 mg) and ethanol (4 mL) was added a 6 M aqueous sodium hydroxide solution (2 mL), followed by heating to reflux for 5 hours. The reaction mixture was cooled to room temperature and water was added thereto, followed by extraction with chloroform. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain 5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (264 mg) as a solid.
Preparation Example 8
To a mixture of tert-butyl (3R)-4-[5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl]-3-methylpiperzine-1-carboxylate (19.9 g) and methanol (60 mL) was added hydrogen chloride (4 M dioxane solution, 180 mL), followed by stirring at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. To the residue was added ethyl acetate (250 mL), followed by stirring at room temperature for 30 minutes. The solid was collected by filtration to obtain N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin -1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamide trihydrochloride (20.1 g) as a solid.
Preparation 9
To a mixture of tert-butyl (3S)-4-(3-ethoxy-3-oxopropyl)-3-methylpiperazine-1-carboxylate (1.2 g) and ethanol (6 mL) was added hydrogen chloride (4 M ethyl acetate solution, 6 mL), followed by stirring at 80° C. for 1.5 hours. The reaction mixture was cooled to room temperature and stirred overnight. The solid was collected by filtration to obtain ethyl 3-[(2S)-2-methylpiperazin-1-yl]propanoate dihydrochloride (995 mg) as a solid.
Preparation Example 10
To a mixture of tert-butyl (2R)-2-ethylpyrrolidine-1-carboxylate (3.4 g) and dioxane (25 mL) was added hydrogen chloride (4 M dioxane solution, 25 mL), followed by stirring at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and to the residue were added diethyl ether, followed by stirring. The solid was collected by filtration to obtain (2R)-2-ethylpyrrolidine hydrochloride (2.1 g) as a solid.
Preparation Example 11
A mixture of {2-acetamido-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl acetate (500 mg), diethylamine (0.3 mL), N,N-diisopropylethylamine (0.7 mL), and N-methylpyrrolidone (5 mL) was stirred at 100° C. for 2 hours. To the reaction mixture was added ethyl acetate, followed by washing with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain N-{5-[(diethylamine)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide (397 mg) as a solid.
Preparation Example 12
To a mixture of {2-acetamide-4-[3-chloro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl acetate (900 mg) and N,N-dimethylformamide (4 mL) were added (2R)-2-methylpyrrolidine (293 mg) and N,N-diisopropylethylamine (0.78 mL), followed by stirring at 110° C. for 30 minutes under microwave irradiation. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain N-(4-[3-chloro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)acetamide (896 mg) as a solid.
Preparation Example 13
A mixture of N-[4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]acetamide (6.0 g), acetic acid (30 mL), a 36% aqueous formaldehyde solution (7.5 mL), and acetic anhydride (9 mL) was stirred at 170° C. for 15 minutes under microwave irradiation. The reaction mixture was concentrated under reduced pressure, and to the residue was added ethyl acetate. The mixture was washed with a saturated aqueous sodium hydrogen carbonate solution, water, and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol) and the obtained solid was mixed with diisopropyl ether. The solid was collected by filtration to obtain {2-acetamido-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl acetate (2.6 g) as a solid.
Preparation Example 14
A mixture of ethyl 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (1.0 g), acetic acid (10 mL), a 37% aqueous formaldehyde solution (1.5 mL), and acetic anhydride (1.8 mL) was stirred at 80° C. for 7 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the residue were added water and a saturated aqueous sodium hydrogen carbonate solution, followed by extraction with chloroform/isopropanol. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol).
The obtained compound and pyridine (10 mL) were mixed, and acetic anhydride (0.9 mL) was added thereto, followed by stirring at room temperature for 30 minutes. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium hydrogen carbonate solution, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl 3-[(2R)-4-(5-{[5-(acetoxymethyl)-4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (566 mg) as a solid.
Preparation Example 15
A mixture of N-{4-[4-methoxy-3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide (3.0 g), 37% aqueous formaldehyde solution (7.2 mL), acetic anhydride (9 mL), and acetic acid (30 mL) was stirred at 100° C. for 5 hours. The reaction mixture was concentrated under reduced pressure, and to the residue was added diisopropyl ether. The solid was collected by filtration to obtain {2-acetamido-4-[4-methoxy-3-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl acetate (2.0 g) as a solid.
Preparation Example 16
A mixture of N-{4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide (2.8 g), acetic acid (20 mL), a 36% aqueous formaldehyde solution (3.6 mL), and acetic anhydride (4.4 mL) was stirred at 170° C. for 30 minutes under microwave irradiation. The reaction mixture was concentrated under reduced pressure, and then the obtained solid was washed with methanol and collected by filtration.
The obtained solid (1.8 g) was mixed with N-methylpyrrolidone (20 mL), (2R)-2-methylpyrrolidine (608 mg), and N,N-diisopropylethylamine (2.5 mL), followed by stirring at 100° C. for 30 minutes. The reaction was cooled to room temperature, and water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)acetamide (1.4 g) as a solid.
Preparation Example 17
N-[4-(4-Chlorothiophen-2-yl)-1,3-thiazol-2-yl]-2,2,2-trifluoroacetamide (5.0 mL), and a 36% aqueous formaldehyde solution (2.5 mL) were mixed, followed by stirring at 60° C. for 1 hour. The reaction mixture was concentrated under reduced pressure and diluted with ethyl acetate. The mixture was washed with a saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. A mixture of the obtained compound, ethanol (50 mL), and a 6 M aqueous sodium hydroxide solution (14 mL) was stirred at 90° C. for 2 hours. The reaction mixture was cooled to room temperature, and water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain 4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]-methyl}-1,3-thiazol-2 -amine (2.7 g) as a solid.
Preparation Example 18
To a mixture of ethyl 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (20 g) and acetic acid (200 mL) were added paraformaldehyde (3.5 g) and (2R)-2-methylpyrrolidine (6.6 g), followed by stirring at 75° C. for 3.5 hours. The reaction mixture was concentrated under reduced pressure. To the residue water added ethyl acetate (250 mL), toluene (125 mL), and water (200 mL), followed by neutralization by the addition of sodium carbonate. The organic layer was separated, the aqueous layer was extracted with ethyl acetate/toluene, the organic layers were dried over anhydrous sodium sulfate, and then amino silica gel (40 g) was added thereto. The mixture was stirred at room temperature for 30 minutes, the insoluble materials were separated by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (19.5 g) as a solid.
Preparation Example 19
4-[3-Fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (2.8 g), pyridine (10 mL), and acetic acid anhydride (4 mL) were mixed, by stirring at 60° C. for 1 hour. The reaction mixture was cooled to room temperature, water was added thereto, and the generated solid was collected by filtration. The obtained solid was washed with methanol and the solid was collected by filtration to obtain N-{4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-triazol-2-yl}acetamide (2.9 g) as a solid.
Preparation Example 20
A mixture of 4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-amine (5.0 g), dichloromethane (100 mL), and triethylamine (5.0 mL) was stirred and ice-cooled, and trifluoroacetic anhydride (5 mL) was added thereto, followed by stirring a room temperature for 1 hour. The reaction mixture was diluted with chloroform, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate). The obtained solid was washed with hexane and the solid was collected by filtration to obtain N-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]-2,2,2-trifluoroacetamide (6.0 g) as a solid.
Preparation Example 21
A mixture of tert-butyl (3S)-4-{5-[(4-[3-fluoro-5(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl ]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazine-1-carboxylate (410 mg), hydrogen chloride (4 M dioxane solution, 4 mL), and methanol (2 mL) was stirred at room temperature for 1 hour. To the reaction mixture was added ethyl acetate, followed by concentration under reduced pressure. A mixture of the obtained compound, N-methylpyrrolidone (6 mL), ethyl 3-bromopropanoate (0.4 mL), and potassium carbonate (683 mg) was stirred at 100° C. for 2 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate. The mixture was washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl 3-[(3S)-4-[5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl]-1,3-thiazol-2-yl)carbamoyl]pyrazine-2-yl}-3-methylpiperazin-1-yl]propanoate 205 mg).
Preparation Example 22
A mixture of tert-butyl (3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazine-1-carboxylate (271 mg), hydrogen chloride (4 M dioxane solution, 4 mL), and methanol (2 mL) was stirred at room temperature for 1 hour. To the reaction mixture was added ethyl acetate, followed by concentration under reduced pressure. A mixture of the residue, N,N-dimethylformamide (4 mL), ethyl bromoacetate (0.05 mL), and N,N-diisopropylethylamine (0.3 mL) was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) and purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl [(3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]acetate (1.54 mg) as a solid.
Preparation Example 23
A mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (1 g), iodoethane (1.2 mL), cesium carbonate (1.9 g), and N,N-dimethylformamide (15 mL) was stirred at 60° C. for 3 hours. The reaction mixture was cooled to room temperature, and water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 1-[4-ethoxy-3-(trifluoromethyl)phenyl]ethanone (1.1 g) as a sold.
Preparation Example 24
To a mixture of 4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-amine (500 mg) and dichloromethane (10 mL) were added 5-chloropyrazine-2-carboxylic acid (530 mg), WSCD.HCl (730 mg), and N,N-dimethyl-4-aminopyridine (100 mg), followed by stirring at 40° C. for 30 minutes. The reaction mixture was cooled to room temperature, and ethyl acetate, water, and a saturated aqueous sodium hydrogen carbonate solution were added thereto. The insoluble materials were separated by filtration over Celite and the filtrate was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. To a mixture of the obtained compound and N-methylpyrrolidone (16 mL) were added ethyl 3-(piperazin-1-yl)propanoate dihydrochloride (1.0 g) and N,N-diisopropylethylamine (3 mL), followed by stirring at 80° C. for 2 hours. The reaction mixture was cooled to room temperature, and water and ethyl acetate were added thereto. The organic layer was washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-ethyl acetate). The obtained compound was washed with diisopropyl ether (4 mL) and hexane (20 mL), and the solid was collected by filtration to obtain ethyl 3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoate (95.4 mg) as a solid.
Preparation Example 25
To a mixture of N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamide trihydrochloride (16.1 g) and N,N-dimethylformamide (400 mL) was added potassium carbonate (11.5 g), followed by stirring at room temperature for 5 minutes. To the reaction mixture was added ethyl bromoacetate (2.65 mL), followed by stirring at room temperature for 1 hour. To the reaction mixture was added ethyl bromoacetate (0.8 mL), followed by stirring at room temperature for 1.5 hours. The reaction mixture was poured into water, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, and anhydrous magnesium sulfate and activated carbon were added thereto. The insoluble materials were separated by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetate (11.0 g) as a solid.
Preparation Example 26
To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethane (1 g) and acetonitrile (10 mL) were added 1-bromopropane (0.9 mL), potassium carbonate (1.7 g), and tetrabutylammonium iodide (180 mg), followed by stirring at room temperature overnight. The insoluble materials were separated by filtration and the filtrate was concentrated under reduced pressure. The reside was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 1-[4-propoxy-3-(trifluoromethyl)phenyl]ethanone (1.2 g) as an oil.
Preparation Example 27
To a mixture of copper iodide (I) (9.4 g) and diethyl ether (180 mL) was added dropwise methyllithium (about 1 M diethyl ether solution, 100 mL) at an internal temperature of 0° C. to 5° C. over 30 minutes, followed by stirring for 15 minutes. To the reaction mixture was added dropwise a solution of tert-butyl (2S)-2-({[(4-methylphenyl)sulfonyl]oxy}methyl)pyrrolidine-1-carboxylate (7.0 g) in dichloromethane (30 mL) at an internal temperature of 5° C. or lower over 20 minutes, followed by stirring at room temperature for 2.5 hours. To the reaction mixture was added dropwise a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain tert-butyl (2R)-2-ethylpyrrolidine-1-carboxylate (3.5 g) as an oil.
Preparation Example 28
A mixture of tert-butyl (2R)-2-methylpiperazine-1-carboxylate (3.0 g), N,N-dimethylformamide (30 mL), ethyl bromoacetate (2 mL), and potassium carbonate (5.0 g) was stirred at room temperature for 1 hour. To the reaction mixture was added ethyl acetate, followed by washing with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol) to obtain tert-butyl (2R)-4-(2-ethoxy-2-oxoethyl)-2-methylpiperazine-1-carboxylate (4.0 g) as an oil.
Preparation Example 29
To a mixture of 5-chloro-N-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]pyrazine-2-carboxamide (25.0 g) and N-methylpyrrolidone (50 mL) were added N,N-diisopropylethylamine (50 mL) and ethyl 3-[(2S)-methylpiperazin-1-yl]propanoate dihydrochloride (21.2 g), followed by stirring at 60° C. for 1.5 hours. The reaction mixture was cooled to room temperature, and ethyl acetate and water were added thereto, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, and anhydrous magnesium sulfate and activated carbon were added thereto. The insoluble materials were separate by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-ethyl acetate). The obtained compound was mixed with diisopropyl ether (40 mL) and hexane (120 mL), followed by stirring at room temperature for 15 minutes. The solid was collected by filtration to obtain 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (29.7 g) as a solid.
Preparation Example 30
To a mixture of 1-[3-fluoro-5-(trifluoromethyl)phenyl]ethanone (78 g) and tetrahyrofuran (625 mL) was added phenyltrimethylammonium tribromide (143 g), followed by stirring at room temperature for 1 hour. The insoluble materials were separated by filtration and the filtrate was concentrated under reduced pressure.
The obtained compound and ethanol (625 mL) were mixed, and thiourea (35 g) was added thereof, followed by stirring at 65° C. to 75° C. for 2 hours. The reaction mixture was ice-cooled, and water (625 mL) was added thereto. The the mixture was added a 1 M sodium hydroxide (600 mL), followed by stirring for 30 minutes. The solid was collected by filtration, and ethanol (30% aqueous, 600 mL) was added thereto and dissolved at 76° C. The obtained solution was cooled to room temperature and stirred overnight. The mixture was ice-cooled and stirred for 2 hours, and then the precipitated solid was collected by filtration to obtain 4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (56.9 g) as a solid.
Preparation Example 31
To a mixture of 1-(4-bromothiophen-2-yl)ethanone (20 g) and N-methylpyrrolidone (400 mL) were added sodium trifluoroacetate (140 g) and copper iodide (I) (100 g), followed by stirring at 200° C. for 2.5 hours. The reaction mixture was cooled to room temperature, water and ethyl acetate were added thereto, and the insoluble materials were separated by filtration over Celite. The organic layer of the filtrate was separated, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) and purified by basic silica gel column chromatography (hexane-ethyl acetate) to obtain 1-[4-(trifluoromethyl) thiophen-2-yl]ethanone (4.1 g) as an oil.
Preparation Example 32
To a mixture of N,6-dimethoxy-N-methyl-5-(trifluoromethyl)nicotinamide (3.7 g) and tetrahydrofuran (40 mL) was added methylmagnesium bromide (3 M tetrahydrofuran solution 7 mL) under ice-cooling, followed by stirring for 1 hour. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain (1-[6-methoxy-5-(trifluoromethyl)pyridin-3-yl]ethanone (3.0 g) as an oil.
Preparation Example 33
A mixture of 1-(3,5-dichloro-4-hydroxyphenyl)ethanone (10.0 g), N,N-dimethylformamide (100 mL), potassium carbonate (8.1 g), and methyl iodide (6.1 mL) was stirred at room temperature overnight. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with 1 M hydrochloride acid and saturated brine, and dried over anhydrous magnesium sulfate. The mixture was filtered using a basic silica gel and the filtrate was concentrated under reduced pressure to obtain 1-(3,5-dichloro-4-methoxyphenyl)ethanone (7.6 g) as a solid.
Preparation Example 34
To a mixture of ethyl 6-methoxy-5-(trifluoromethyl)nicotinate (5.5 g) and ethanol (40 mL) were added a 3 M aqueous sodium hydroxide solution (40 mL), followed by stirring at 60° C. for 30 minutes. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the residue was added 1 M hydrochloric acid (120 mL) and the mixture was stirred for 1 hour. The precipitated solid was collected by filtration to obtain 6-methoxy-5-(trifluoromethyl)nicotinic acid (4.4 g) as a solid.
Preparation Example 35
A mixture of 5-bromo-2-methoxy-3-(trifluoromethyl) pyridine (7.8 g), palladium acetate (II) (170 mg), 1,1′-bis(diphenylphosphino)ferrence (840 mg), N,N-diisopropylethylamine (10 mL), ethanol (80 mL), and N,N-dimethylformamide (80 mL) was stirred at 90° C. for 19 hours under a carbon monoxide atmosphere. The reaction mixture was cooled to room temperature, and poured into water (500 mL) and ethyl acetate (500 mL), followed by stirring for 30 minutes. The organic layer was separated, washed with water and saturated brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain ethyl 6-methoxy-5-(trifluoromethyl)nicotinate (5.5 g) as a solid.
Preparation Example 36
2-Methoxy-3-(trifluoromethyl)pyridine (8 g), 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (17 g), and trifluoroacetic acid (32 mL) were mixed, followed by stirring at room temperature for 22 hours. The reaction mixture was concentrated under reduced pressure, and to the residue was added diisopropyl ether. The precipitated solid was separated by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 5-bromo-2-methoxy-3-(trifluoromethyl)pyridine (9.4 g) as an oil.
Preparation Example 37
To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (1 g) and tetrahydrofuran (10 mL) were added 2-propanol (0.46 mL), a 40% diethylazodicarboxylate solution in toluene (2.3 mL) and triphenylphosphine (1.6 g), followed by stirring at room temperature overnight. The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 1-[4-isopropoxy-3-(trifluoromethyl)phenyl]ethanone (1.0 g) as an oil.
Preparation Example 38
A mixture of 1-[4-chloro-3-(trifluoromethyl)phenyl]ethanone (1.0 g), cyclopropylboronic acid (780 mg), dicyclohexyl (2′,6′-dimethoxybiphenyl-2-yl)phosphine (185 mg), tripotassium phosphate (3.0 g), palladium acetate (II), (51 mg), toluene (10 mL), and water (1 mL was stirred at 100° C. for 3 hours under an argon atmosphere. The reaction mixture was cooled to room temperature, ethyl acetate and water were added thereto, and the insoluble materials were separated by filtration. The filtrate was extracted with ethyl acetate and the organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 1-[4-cyclopropyl-3-(trifluoromethyl)phenyl]ethanone (10 ) as an oil.
Preparation Example 39
To a mixture of 1-(4-bromothiophen-2-yl)ethanone (9.4 g), toluene (200 mL) and water (100 mL) were added cyclopropylboronic acid (12.0 g), tetrakis(triphenylphosphine) palladium (0) (5.34 g), cesium carbonate (73.6 g), and tri-tert-butylphosphine (2.3 mL), followed by stirring at 80° C. for 3 hours. The reaction mixture was filtered over Celite, and to the filtrate were added water and diethyl ether. The organic layer was separated, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain 1-(4-cyclopropylthiophen-2-yl)ethanone (6.7 g ) as an oil.
Preparation Example 40
A mixture of 3-bromo-5-(trifluoromethyl)benzoic acid (10.0 g), thionylchloride (40 mL), and N,N-dimethylformamide (1 droplet) was stirred at 802 C. for 2 hours. The reaction mixture was concentrated under reduced pressure, followed by carrying out a concentration operation with toluene twice and then drying under reduced pressure.
To a mixture of toluene (150 mL) and magnesium chloride (3.6 g) were added dimethyl malonate (5.1 mL) and triethylamine (12 mL), followed by stirring at room temperature for 1.5 hours. To the reaction mixture was first added dropwise a mixture of the obtained compound and toluene (50 mL) under stirring, followed by stirring at room temperature for 18 hours. To the reaction mixture was added 6 M hydrochloric acid (50 mL), and then water (300 mL) was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was mixed with dimethylsulfoxide (50 mL) and water (5 mL), followed by stirring at 160° C. for 1 hour. The reaction mixture was cooled to room temperature, and water (300 mL) was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain 1-[3-bromo-5-(trifluoromethyl)phenyl]ethanone (10.0 g) as an oil.
Preparation Example 41
To a mixture of zinc powder (2.0 g), cobalt bromide (II) (600 mg), and acetonitrile (30 mL) was added trifluoroacetic acid (0.15 mL) under an argon atmosphere, followed by stirring at room temperature for 15 minutes. To the reaction mixture were added 5-bromo-1-fluoro-2-methoxy-3-(trifluoromethyl)benzene (5.0 g) and acetic anhydride (2.1 mL), followed by stirring at room temperature for 17 hours. To the reaction mixture was added 1 M hydrochloric acid (30 mL), followed by extraction with diethyl ether. The organic layer was washed with water and saturated with brine, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-diethyl ether) to obtain 1-[3-fluoro-4-methoxy-5-(trifluoromethyl)phenyl]ethanone (1.6 g) as an oil.
Preparation Example 42
To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (3.0 g), N,N-dimethylformamide (36 mL), and water (3.6 mL) were added sodium chloro(difluoro)acetate (5.8 g) and cesium carbonate (7.2 g), followed by stirring at 100° C. for 3 hours. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate). To a mixture of the obtained compound (3.8 g) and tetrahydrofuran (50 mL) was added phenyltrimethylammonium tribromide (5.7 g), followed by stirring at room temperature for 45 minutes. The precipitated insoluble materials were separated by filtration and the filtrate was concentrated under reduced pressure. To a mixture of the residue and ethanol (50 mL) was added thiourea (1.5 g), followed by stirring at 80° C. for 2 hours. The reaction mixture was cooled to room temperature, and water (30 mL) and a 1 M aqueous sodium hydroxide solution (30 mL) were added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. To the residue was added diisopropyl ether and hexane, and the generated solid was collected by filtration to obtain 4-[4-(difluoromethoxy)-3-(trifluoromethyl)phenyl]-3-thiazol-2-amine (3.5 g) as a solid.
Preparation Example 43
To a mixture of 5-chloro-N-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)pyrazine-2-carboxamide (407 mg) and N-methylpyrrolidone (6 mL) were added tert-butyl (3R)-3-methylpiperazine-1-carboxylate (400 mg) and N,N-diisopropylethylamine (0.7 mL), followed by stirring at 80° C. for 4 hours. The reaction mixture was cooled to room temperature, and water was added thereto, followed by extraction with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate).
A mixture of the obtained compound, hydrogen chloride (4 M dioxane solution, 6 mL), and methanol (2 mL) was stirred at room temperature for 4 hours. To the reaction mixture was added ethyl acetate (20 mL), and the solid was collected by filtration to obtain N-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamide trihydrochloride (623 mg) as a solid.
Preparation Example 44
To a mixture of tert-butyl (2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate (17 g), triethylamine (17.7 mL), 1-methyl-1H-imidazole (10.1 mL), and dichloromethane (255 mL) was added p-toluenesulfonyl chloride (17.7 g) under ice-cooling, followed by stirring at the same temperature for 1 hour. To the reaction mixture was added water, followed by extraction with dichloromethane. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain tert-butyl (2S)-2-({[(4-methylphenyl)sulfonyl]oxy}methyl)pyrrolidine-1-carboxylate (29.54 g) as an oil.
Preparation Example 45
A mixture of tert-butyl (3S)-3-methylpiperazine-1-carboxylate (5 g), ethyl acrylate (7.2 mL), and ethanol (15 mL) was heated and refluxed for 24 hours. The reaction mixture was concentrated under reduced pressure, and to the residue was added diethyl ether, followed by extraction with 1 M hydrochloric acid. The aqueous layer was alkalified to pH 8 by the addition of a 1 M aqueous sodium hydroxide solution and sodium hydrogen carbonate, and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol) to obtain tert-butyl (3 S)-4-(3-ethoxy-3-oxopropyl)-3-methylpiperazine-1-carboxylate (7.5 g) as an oil.
Example 1
To a mixture of ethyl 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (10.2 g), tetrahydrofuran (50 mL), and ethanol (50 mL) was added a 1 M aqueous sodium hydroxide solution (50 mL), followed by stirring at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (50 mL) and water (100 mL) were added thereto, followed by extraction with chloroform. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol) to obtain a solid (6.0 g) of 3-[(2S)-4-(5{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid.
To a mixture of the obtained solid and tetrahydrofuran (100 mL) was added hydrogen chloride (4 M dioxane solution, 12 mL), and the mixture was concentrated under reduced pressure. To the residue were added acetonitrile (200 mL) and water (12 mL), followed by stirring at 70° C. for 15 minutes, and then cooling at room temperature. To the mixture was added acetonitrile (100 mL), followed by stirring at room temperature for 1 hour. The solid was collected by filtration and dried to obtain 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid dihydrochloride (6.7 g) as a solid.
Example 2
Under an argon gas flow, to a mixture of ethyl 3-(4-{5-[(4-[3-bromo-5-(trifluoromethyl)phenyl]-5-{[(2S)-2-isopropylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoate (660 mg), zinc powder (30 mg), biphenyl-2-yl(di-tert-butyl)phosphine (60 mg), and N,N-dimethylacetamide (13 mL) were added zinc cyanide (160 mg) and palladium trifluoroacetate (II) (30 mg), followed by stirring at 100° C. for 1 hour. The reaction mixture was cooled to room temperature, and ethyl acetate was added thereto. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound (401 mg), ethanol (5 mL), and tetrahydrofuran (5 mL) was added a 1 M aqueous sodium hydroxide solution (3 mL), followed by stirred at 50° C. for 30 minutes. The reaction mixture was concentrated under reduced pressure and the residue was purified by ODS column chromatography (acetonitrile-water). The obtained solid was mixed with hexane (20 mL) and diethyl ether (4 mL), and the solid was collected by filtration to obtain sodium 3-(4-{5-[(4-[3-cyano-5-(trifluoromethyl)phenyl]-5-{[(2S)-2-isopropylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoate (149 mg) as a solid.
Example 3
To a mixture of 5-chloro-N-(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide (300 mg) and N-methylpyrrolidone (6 mL) were added ethyl 3-[(3R)-3-methylpiperazin-1-yl]propanoate dihydrochloride (500 mg) and N,N-diisopropylethylamine (0.64 mL), followed by stirring at 90° C. for 2 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound, ethanol (6mL), and tetrahydrofuran (6 mL) was added a 1 M aqueous sodium hydroxide solution (3.5 mL), followed by stirring at 60° C. for 30 minutes. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by ODS column chromatography (acetonitrile—0.1% aqueous formic acid solution) to obtain a solid (204 mg). To a mixture of the obtained solid and ethyl acetate was added hydrogen chloride (4 M ethyl acetate solution, 0.25 mL). The reaction mixture was concentrated under reduced pressure to obtain 3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid dihydrochloride (155 mg) as a solid.
Example 4
To a mixture of 5-chloro-N-(5-{[(2R)-2-methylpiperidin-1-yl]methyl}-4-[3-methyl-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide (300 mg) and N-methylpyrrolidone (6 mL) were added ethyl 3-(piperazin-1-yl)propanoate dihydrochloride (250 mg) and N,N-diisopropylethylamine (0.7 mL), followed by stirring at 80° C. for 2 hours. The reaction mixture was cooled to room temperature, and water and ethyl acetate were added thereto. The organic layer was separated, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained residue, ethanol (5 mL), and tetrahydrofuran (5 mL) was added a 1 M aqueous sodium hydroxide solution (3 mL), followed by stirring at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by ODS column chromatography (acetonitrile-water) to obtain a solid (298 mg). The obtained solid was mixed with hexane (10 mL) and diethyl ether (2 mL), and the solid was collected by filtration to obtain sodium 3-(4-{5-[(5-{[(2R)-2-methylpiperidin-1-yl]methyl}-4-[3-methyl]-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazine-2-yl}piperazin-1-yl)propanoate (284 mg) as a solid.
Example 5
A mixture of ethyl 3-[(2R)-4-(5-{[5-(acetoxymethyl)-4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate (200 mg), dimethylamine (2M tetrahydrofuran solution, 2 mL), and N-methylpyrrolidone (4 mL) was stirred at 80° C. for 3 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate) and purified by silica gel column chromatography (hexane-ethyl acetate). The obtained compound was mixed with ethanol (2 mL) and tetrahydrofuran (2 mL), and a 1 M aqueous sodium hydroxide solution (1 mL) was added thereto, followed by stirring at room temperature for 1 hour. To the reaction mixture was added 1 M hydrochloric acid (1 mL) and water, the mixture was extracted with chloroform/isopropanol, and the organic layer was washed with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. To a mixture of the obtained compound and ethyl acetate was added hydrogen chloride (4 M ethyl acetate solution, 1 mL). The reaction mixture was concentrated under reduced pressure, and to the residue was added ethyl acetate. The solid was collected by filtration to obtain 3-{(2R)-4-[5-({4-(4-chlorothiophen-2-yl)-5-[(dimethylamino)methyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid dihydrochloride (33 mg) as a solid.
Example 6
A mixture of ethyl 3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoate (400 mg), (2R)-2-methylpyrrolidine (273 mg), a 36% aqueous formaldehyde solution (0.5 mL), and acetic acid (8 mL) was stirred at 60° C. for 1.5 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the residue was added a saturated aqueous sodium hydrogen carbonate solution, followed by extraction with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound (452 mg), ethanol (4 mL), and tetrahydrofuran (4 mL) was added a 1 M aqueous sodium hydroxide solution (4 mL), followed by stirring at 50° C. for 1 hour. The reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (4 mL) and water were added thereto. The mixture was extracted from chloroform/isopropanol/tetrahydrofuran, and the organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. To a mixture of the obtained compound and tetrahydrofuran (20 mL) was added hydrogen chloride (4 M dioxane solution, 2 mL). The mixture was concentrated under reduced pressure, and to the residue was added diethyl ether (20 mL). The solid was collected by filtration to obtain 3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoic acid trihydrochloride (440 mg) as a solid.
Example 7
To a mixture of N-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamide trihydrochloride (300 mg) and N,N-dimethylformamide (5 mL) were added potassium carbonate (300 mg) and ethyl 3-bromopropanoate (0.25 mL), followed by stirring at 60° C. for 1.5 hours. Thereafter, to the reaction mixture were added potassium carbonate (300 mg) and ethyl 3-bromopropanoate (0.25 mL), followed by stirring at 60° C. for 1.5 hours. Again, to the reaction mixture were added potassium carbonate (300 mg) and ethyl 3-bromopropanoate (0.25 mL), followed by stirring at 60° C. for 1 hour. The reaction mixture was cooled to room temperature, and water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound (151 mg), tetra hydrofuran (2 mL), and ethanol (2 mL) was added a 1 M aqueous sodium hydroxide solution (1 mL), followed by stirring at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (1 mL) and water (15 mL) were added thereto, followed by extracted with chloroform/isopropanol. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. To a mixture of the obtained compound and tetrahydrofuran (10 mL was added hydrogen chloride (4 M dioxane solution, 2 mL). The reaction mixture was concentrated under reduced pressure, and to the residue was added diethyl ether. The solid was collected by filtration to obtain 3-[(3R)-4-{5-[(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid trihydrochloride (142 mg) as a solid.
Example 8
To a mixture of N-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamide trihydrochloride (381 mg) and N,N-dimethylformamide (8 mL) was added potassium carbonate (300 mg), followed by stirring at room temperature for 10 minutes. To the reaction mixture was added ethyl bromoacetate (0.09 mL), followed by stirring at room temperature for 1.5 hours. To the reaction mixture was added ethyl bromoacetate (0.09 mL), followed by stirring at room temperature for 30 minutes. To the reaction mixture was added water, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by basic silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound (211 mg), tetrahydrofuran (3 mL), and ethanol (3 mL) was added a 1 M aqueous sodium hydroxide solution (1.5 mL), followed by stirring at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (1.5 mL) and water (15 mL) were added thereto, followed by extraction with chloroform/isopropanol. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was mixed with tetrahydrofuran (10 mL), and hydrogen chloride (4 M dioxane solution, 2 mL) was added thereto. The mixture was concentrated under reduced pressure, and to the residue was added diethyl ether. The solid was collected by filtration to obtain [(3R)-4-{5-[(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid trihydrochloride (185 mg).
Example 9
To a mixture of 5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (820 mg), triethylamine (2 mL), and cyclopentylmethyl ether (16 mL) was added 5-chloropyrazine-2-carbonylchloride (590 mg), followed by stirring at room temperature for 20 hours. To the reaction mixture was added water (500 mL), followed by extraction with chloroform. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain a solid (1.0 g). To a mixture of the obtained compound (200 mg) and N-methylpyrrolidone (4 mL) were added ethyl 3-[(2R)-2-methylpiperazin-1-yl]propanoate dihydrochloride (168 mg) and N,N-diisopropylethylamine (0.5 mL), followed by stirring at 80° C. for 2 hours. The reaction mixture was cooled to room temperature, and water and ethyl acetate were added thereto. The organic layer was separated, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained compound was purified by silica gel column chromatography (hexane-ethyl acetate).
To a mixture of the obtained compound (249 mg), ethanol (4 mL), and tetrahydrofuran (4 mL) was added a 1 M aqueous sodium hydroxide solution (2 mL), followed by stirring at 50° C. for 30 minutes. The reaction mixture was cooled to room temperature, and 1 M hydrochloric acid (2 mL) and water (20 mL) were added thereto. The mixture was extracted with chloroform/isopropanol, and the organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was mixed with tetrahydrofuran (10 mL), and hydrogen chloride (4 M dioxane solution, 2 mL) was added thereto. The mixture was concentrated under reduced pressure, and to the residue was added diethyl ether. The solid was collected by filtration to obtain 3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazine-2-yl]-2-methylpiperazin-1-yl}propanoic acid dihydrochloride (251 mg) as a solid.
Example 144
3-[(2S)-4-(5-{[4-(4-Chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid (500 mg) and maleic acid (148 mg) were dissolved in 2-butanone (0.5 mL) and dimethylsulfoxide (0.5 mL) under stirring at 60° C. To the solution was added 2-butanone (4.0 mL), followed by stirring at 60° C. for 30 minutes. Thereafter, the mixture was left to be slowly cooled to room temperature and stirred at room temperature for 16 hours. The precipitated solid was collected by filtration and dried under reduced pressure to obtain 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid dimalate dimaleate (378 mg) as a white crystal.
The crystals obtained in the present Examples have peaks of powder X-ray diffraction at 2θ (°) 5.7, 6.6, 10.5, 12.0, 13.3, 15.8, 16.6, 17.3, 19.0, and 26.2.
The compounds of Preparation Examples and Examples shown in Tables below were produced in the same manner as the methods in Preparation Examples or Examples as described above.
TABLE 5
PEx Structure
1
Figure USRE049111-20220621-C00009
2
Figure USRE049111-20220621-C00010
3
Figure USRE049111-20220621-C00011
4
Figure USRE049111-20220621-C00012
5
Figure USRE049111-20220621-C00013
TABLE 6
PEx Structure
6
Figure USRE049111-20220621-C00014
7
Figure USRE049111-20220621-C00015
8
Figure USRE049111-20220621-C00016
9
Figure USRE049111-20220621-C00017
10
Figure USRE049111-20220621-C00018
TABLE 7
PEx Structure
11
Figure USRE049111-20220621-C00019
12
Figure USRE049111-20220621-C00020
13
Figure USRE049111-20220621-C00021
14
Figure USRE049111-20220621-C00022
15
Figure USRE049111-20220621-C00023
TABLE 8
PEx Structure
16
Figure USRE049111-20220621-C00024
17
Figure USRE049111-20220621-C00025
18
Figure USRE049111-20220621-C00026
19
Figure USRE049111-20220621-C00027
20
Figure USRE049111-20220621-C00028
TABLE 9
PEx Structure
21
Figure USRE049111-20220621-C00029
22
Figure USRE049111-20220621-C00030
23
Figure USRE049111-20220621-C00031
24
Figure USRE049111-20220621-C00032
25
Figure USRE049111-20220621-C00033
TABLE 10
PEx Structure
26
Figure USRE049111-20220621-C00034
27
Figure USRE049111-20220621-C00035
28
Figure USRE049111-20220621-C00036
29
Figure USRE049111-20220621-C00037
30
Figure USRE049111-20220621-C00038
31
Figure USRE049111-20220621-C00039
32
Figure USRE049111-20220621-C00040
TABLE 11
PEx Structure
33
Figure USRE049111-20220621-C00041
34
Figure USRE049111-20220621-C00042
35
Figure USRE049111-20220621-C00043
36
Figure USRE049111-20220621-C00044
37
Figure USRE049111-20220621-C00045
38
Figure USRE049111-20220621-C00046
39
Figure USRE049111-20220621-C00047
40
Figure USRE049111-20220621-C00048
TABLE 12
PEx Structure
41
Figure USRE049111-20220621-C00049
42
Figure USRE049111-20220621-C00050
43
Figure USRE049111-20220621-C00051
44
Figure USRE049111-20220621-C00052
45
Figure USRE049111-20220621-C00053
46
Figure USRE049111-20220621-C00054
TABLE 13
PEx Structure
47
Figure USRE049111-20220621-C00055
48
Figure USRE049111-20220621-C00056
49
Figure USRE049111-20220621-C00057
50
Figure USRE049111-20220621-C00058
TABLE 14
PEx Structure
51
Figure USRE049111-20220621-C00059
52
Figure USRE049111-20220621-C00060
53
Figure USRE049111-20220621-C00061
54
Figure USRE049111-20220621-C00062
TABLE 15
PEx Structure
55
Figure USRE049111-20220621-C00063
56
Figure USRE049111-20220621-C00064
57
Figure USRE049111-20220621-C00065
58
Figure USRE049111-20220621-C00066
TABLE 16
PEx Structure
59
Figure USRE049111-20220621-C00067
60
Figure USRE049111-20220621-C00068
61
Figure USRE049111-20220621-C00069
62
Figure USRE049111-20220621-C00070
TABLE 17
PEx Structure
63
Figure USRE049111-20220621-C00071
64
Figure USRE049111-20220621-C00072
65
Figure USRE049111-20220621-C00073
66
Figure USRE049111-20220621-C00074
TABLE 18
PEx Structure
67
Figure USRE049111-20220621-C00075
68
Figure USRE049111-20220621-C00076
69
Figure USRE049111-20220621-C00077
70
Figure USRE049111-20220621-C00078
71
Figure USRE049111-20220621-C00079
TABLE 20
PEx Structure
77
Figure USRE049111-20220621-C00080
78
Figure USRE049111-20220621-C00081
79
Figure USRE049111-20220621-C00082
80
Figure USRE049111-20220621-C00083
81
Figure USRE049111-20220621-C00084
TABLE 19
PEx Structure
72
Figure USRE049111-20220621-C00085
73
Figure USRE049111-20220621-C00086
74
Figure USRE049111-20220621-C00087
75
Figure USRE049111-20220621-C00088
76
Figure USRE049111-20220621-C00089
TABLE 21
PEx Structure
82
Figure USRE049111-20220621-C00090
83
Figure USRE049111-20220621-C00091
84
Figure USRE049111-20220621-C00092
85
Figure USRE049111-20220621-C00093
86
Figure USRE049111-20220621-C00094
TABLE 22
PEx Structure
87
Figure USRE049111-20220621-C00095
88
Figure USRE049111-20220621-C00096
89
Figure USRE049111-20220621-C00097
90
Figure USRE049111-20220621-C00098
TABLE 23
PEx Structure
91
Figure USRE049111-20220621-C00099
92
Figure USRE049111-20220621-C00100
93
Figure USRE049111-20220621-C00101
94
Figure USRE049111-20220621-C00102
TABLE 24
PEx Structure
95
Figure USRE049111-20220621-C00103
96
Figure USRE049111-20220621-C00104
97
Figure USRE049111-20220621-C00105
98
Figure USRE049111-20220621-C00106
TABLE 25
PEx Structure
 99
Figure USRE049111-20220621-C00107
100
Figure USRE049111-20220621-C00108
101
Figure USRE049111-20220621-C00109
102
Figure USRE049111-20220621-C00110
TABLE 26
PEx Structure
103
Figure USRE049111-20220621-C00111
104
Figure USRE049111-20220621-C00112
105
Figure USRE049111-20220621-C00113
106
Figure USRE049111-20220621-C00114
PEx Structure
107
Figure USRE049111-20220621-C00115
108
Figure USRE049111-20220621-C00116
109
Figure USRE049111-20220621-C00117
110
Figure USRE049111-20220621-C00118
111
Figure USRE049111-20220621-C00119
TABLE 28
PEx Structure
112
Figure USRE049111-20220621-C00120
113
Figure USRE049111-20220621-C00121
114
Figure USRE049111-20220621-C00122
115
Figure USRE049111-20220621-C00123
TABLE 29
PEx Structure
116
Figure USRE049111-20220621-C00124
117
Figure USRE049111-20220621-C00125
118
Figure USRE049111-20220621-C00126
119
Figure USRE049111-20220621-C00127
120
Figure USRE049111-20220621-C00128
TABLE 30
PEx Structure
121
Figure USRE049111-20220621-C00129
122
Figure USRE049111-20220621-C00130
123
Figure USRE049111-20220621-C00131
124
Figure USRE049111-20220621-C00132
125
Figure USRE049111-20220621-C00133
126
Figure USRE049111-20220621-C00134
TABLE 31
PEx Structure
127
Figure USRE049111-20220621-C00135
128
Figure USRE049111-20220621-C00136
129
Figure USRE049111-20220621-C00137
130
Figure USRE049111-20220621-C00138
TABLE 32
PEx Structure
131
Figure USRE049111-20220621-C00139
132
Figure USRE049111-20220621-C00140
133
Figure USRE049111-20220621-C00141
134
Figure USRE049111-20220621-C00142
TABLE 33
PEx Structure
135
Figure USRE049111-20220621-C00143
136
Figure USRE049111-20220621-C00144
137
Figure USRE049111-20220621-C00145
138
Figure USRE049111-20220621-C00146
TABLE 34
PEx Structure
139
Figure USRE049111-20220621-C00147
140
Figure USRE049111-20220621-C00148
141
Figure USRE049111-20220621-C00149
142
Figure USRE049111-20220621-C00150
143
Figure USRE049111-20220621-C00151
TABLE 35
PEx Structure
144
Figure USRE049111-20220621-C00152
145
Figure USRE049111-20220621-C00153
146
Figure USRE049111-20220621-C00154
147
Figure USRE049111-20220621-C00155
TABLE 36
PEx Structure
148
Figure USRE049111-20220621-C00156
149
Figure USRE049111-20220621-C00157
150
Figure USRE049111-20220621-C00158
151
Figure USRE049111-20220621-C00159
152
Figure USRE049111-20220621-C00160
TABLE 37
PEx Structure
153
Figure USRE049111-20220621-C00161
154
Figure USRE049111-20220621-C00162
155
Figure USRE049111-20220621-C00163
156
Figure USRE049111-20220621-C00164
157
Figure USRE049111-20220621-C00165
TABLE 38
PEx Structure
158
Figure USRE049111-20220621-C00166
159
Figure USRE049111-20220621-C00167
160
Figure USRE049111-20220621-C00168
161
Figure USRE049111-20220621-C00169
162
Figure USRE049111-20220621-C00170
163
Figure USRE049111-20220621-C00171
TABLE 39
PEx Structure
164
Figure USRE049111-20220621-C00172
165
Figure USRE049111-20220621-C00173
166
Figure USRE049111-20220621-C00174
167
Figure USRE049111-20220621-C00175
168
Figure USRE049111-20220621-C00176
TABLE 40
PEx Structure
169
Figure USRE049111-20220621-C00177
170
Figure USRE049111-20220621-C00178
171
Figure USRE049111-20220621-C00179
172
Figure USRE049111-20220621-C00180
173
Figure USRE049111-20220621-C00181
TABLE 41
PEx Structure
174
Figure USRE049111-20220621-C00182
175
Figure USRE049111-20220621-C00183
176
Figure USRE049111-20220621-C00184
177
Figure USRE049111-20220621-C00185
TABLE 42
PEx Structure
178
Figure USRE049111-20220621-C00186
179
Figure USRE049111-20220621-C00187
180
Figure USRE049111-20220621-C00188
181
Figure USRE049111-20220621-C00189
182
Figure USRE049111-20220621-C00190
TABLE 43
PEx Structure
183
Figure USRE049111-20220621-C00191
184
Figure USRE049111-20220621-C00192
185
Figure USRE049111-20220621-C00193
186
Figure USRE049111-20220621-C00194
187
Figure USRE049111-20220621-C00195
TABLE 44
PEx Structure
188
Figure USRE049111-20220621-C00196
189
Figure USRE049111-20220621-C00197
190
Figure USRE049111-20220621-C00198
191
Figure USRE049111-20220621-C00199
192
Figure USRE049111-20220621-C00200
193
Figure USRE049111-20220621-C00201
TABLE 45
PEx Structure
194
Figure USRE049111-20220621-C00202
195
Figure USRE049111-20220621-C00203
196
Figure USRE049111-20220621-C00204
197
Figure USRE049111-20220621-C00205
198
Figure USRE049111-20220621-C00206
199
Figure USRE049111-20220621-C00207
TABLE 46
PEx Structure
200
Figure USRE049111-20220621-C00208
201
Figure USRE049111-20220621-C00209
202
Figure USRE049111-20220621-C00210
203
Figure USRE049111-20220621-C00211
204
Figure USRE049111-20220621-C00212
205
Figure USRE049111-20220621-C00213
TABLE 47
PEx Structure
206
Figure USRE049111-20220621-C00214
207
Figure USRE049111-20220621-C00215
208
Figure USRE049111-20220621-C00216
209
Figure USRE049111-20220621-C00217
210
Figure USRE049111-20220621-C00218
211
Figure USRE049111-20220621-C00219
212
Figure USRE049111-20220621-C00220
TABLE 48
PEx Structure
213
Figure USRE049111-20220621-C00221
214
Figure USRE049111-20220621-C00222
215
Figure USRE049111-20220621-C00223
216
Figure USRE049111-20220621-C00224
217
Figure USRE049111-20220621-C00225
TABLE 49
PEx Structure
218
Figure USRE049111-20220621-C00226
219
Figure USRE049111-20220621-C00227
220
Figure USRE049111-20220621-C00228
221
Figure USRE049111-20220621-C00229
222
Figure USRE049111-20220621-C00230
TABLE 50
PEx Structure
223
Figure USRE049111-20220621-C00231
224
Figure USRE049111-20220621-C00232
225
Figure USRE049111-20220621-C00233
226
Figure USRE049111-20220621-C00234
TABLE 51
PEx Structure
227
Figure USRE049111-20220621-C00235
228
Figure USRE049111-20220621-C00236
229
Figure USRE049111-20220621-C00237
230
Figure USRE049111-20220621-C00238
231
Figure USRE049111-20220621-C00239
TABLE 52
PEx Structure
232
Figure USRE049111-20220621-C00240
233
Figure USRE049111-20220621-C00241
234
Figure USRE049111-20220621-C00242
235
Figure USRE049111-20220621-C00243
TABLE 53
PEx Structure
236
Figure USRE049111-20220621-C00244
237
Figure USRE049111-20220621-C00245
238
Figure USRE049111-20220621-C00246
239
Figure USRE049111-20220621-C00247
TABLE 54
PEx Structure
240
Figure USRE049111-20220621-C00248
241
Figure USRE049111-20220621-C00249
242
Figure USRE049111-20220621-C00250
243
Figure USRE049111-20220621-C00251
TABLE 55
PEx Structure
244
Figure USRE049111-20220621-C00252
245
Figure USRE049111-20220621-C00253
246
Figure USRE049111-20220621-C00254
247
Figure USRE049111-20220621-C00255
TABLE 56
PEx Structure
248
Figure USRE049111-20220621-C00256
249
Figure USRE049111-20220621-C00257
250
Figure USRE049111-20220621-C00258
251
Figure USRE049111-20220621-C00259
TABLE 57
PEx Structure
252
Figure USRE049111-20220621-C00260
253
Figure USRE049111-20220621-C00261
254
Figure USRE049111-20220621-C00262
255
Figure USRE049111-20220621-C00263
256
Figure USRE049111-20220621-C00264
257
Figure USRE049111-20220621-C00265
TABLE 58
PEx Structure
258
Figure USRE049111-20220621-C00266
259
Figure USRE049111-20220621-C00267
260
Figure USRE049111-20220621-C00268
261
Figure USRE049111-20220621-C00269
262
Figure USRE049111-20220621-C00270
263
Figure USRE049111-20220621-C00271
264
Figure USRE049111-20220621-C00272
TABLE 59
PEx Structure
265
Figure USRE049111-20220621-C00273
266
Figure USRE049111-20220621-C00274
267
Figure USRE049111-20220621-C00275
268
Figure USRE049111-20220621-C00276
269
Figure USRE049111-20220621-C00277
270
Figure USRE049111-20220621-C00278
271
Figure USRE049111-20220621-C00279
TABLE 60
PEx Structure
272
Figure USRE049111-20220621-C00280
273
Figure USRE049111-20220621-C00281
274
Figure USRE049111-20220621-C00282
275
Figure USRE049111-20220621-C00283
276
Figure USRE049111-20220621-C00284
277
Figure USRE049111-20220621-C00285
278
Figure USRE049111-20220621-C00286
TABLE 61
PEx Structure
279
Figure USRE049111-20220621-C00287
280
Figure USRE049111-20220621-C00288
281
Figure USRE049111-20220621-C00289
282
Figure USRE049111-20220621-C00290
283
Figure USRE049111-20220621-C00291
TABLE 62
PEx Structure
284
Figure USRE049111-20220621-C00292
285
Figure USRE049111-20220621-C00293
TABLE 63
PEx PSyn Data
 1 PEx1 ESI+: 500, 502
 2 PEx2 APCI/ESI+: 488
 3 PEx3 NMR-DMSO-d6: 7.55 (1H, d, J = 1.5 Hz), 7.60 (1H, d,
J = 1.5 Hz), 7.76 (1H, s), 8.98 (1H, d, J = 1.3 Hz), 9.15
(1H, d, J = 1.3 Hz), 12.68 (1H, brs)
 4 PEx4 ESI+: 265
 5 PEx5 ESI+: 360
 6 PEx6 ESI+: 374
 7 PEx7 ESI+: 348
 8 PEx8 ESI+: 564
 9 PEx9 ESI+: 201
10 PEx10 ESI+: 100
11 PEx11 ESI+: 390
12 PEx12 ESI+: 418, 420
13 PEx13 ESI+: 377
14 PEx14 ESI+: 593, 585
15 PEx15 ESI+: 389
16 PEx16 ESI+: 402
17 PEx17 ESI+: 328
18 PEx18 ESI+: 618
19 PEx19 ESI+: 305
20 PEx20 ESI+: 313, 315
21 PEx21 ESI+: 664
22 PEx22 ESI+: 604
23 PEx23 ESI+: 233
24 PEx24 ESI+: 501
25 PEx25 ESI+: 650
26 PEx26 ESI+: 247
27 PEx27 ESI+: 200
28 PEx28 ESI+: 287
29 PEx29 ESI+: 521, 523
30 PEx30 ESI+: 263
31 PEx31 CI+: 195
32 PEx32 ESI+: 220
TABLE 64
PEx PSyn Data
33 PEx33 ESI+: 219
34 PEx34 ESI+: 222
35 PEx35 ESI+: 250
36 PEx36 CI+: 256, 258
37 PEx37 ESI+: 247
38 PEx38 EI: 228
39 PEx39 APCI/ESI+: 167
40 PEx40 EI: 266, 268
41 PEx41 ESI+: 237
42 PEx42 ESI+: 311
43 PEx43 ESI+: 590
44 PEx44 ESI+: 378 [M + Na]+
45 PEx45 ESI+: 301
46 PEx1 ESI+: 512, 514
47 PEx1 ESI+: 512
48 PEx1 ESI+: 454
49 PEx1 ESI+: 512
50 PEx1 ESI+: 526, 528
51 PEx1 ESI+: 540, 542
52 PEx1 ESI+: 540, 542
53 PEx1 ESI+: 554, 556
54 PEx1 ESI+: 516, 518
55 PEx1 ESI+: 500
56 PEx1 ESI+: 514, 516
57 PEx1 ESI+: 516, 518
58 PEx1 ESI+: 522
59 PEx1 ESI+: 496, 498
60 PEx1 ESI+: 522, 524
61 PEx1 ESI+: 536, 538
62 PEx1 ESI+: 496, 498
63 PEx1 ESI+: 540, 542
64 PEx1 ESI+: 530
65 PEx1 ESI+: 496
66 PEx1 ESI+: 548
TABLE 65
PEx PSyn Data
67 PEx1 ESI+: 540
68 PEx2 ESI+: 468
69 PEx2 ESI+: 454, 456
70 PEx2 ESI+: 560, 562
71 PEx2 ESI+: 510, 512
72 PEx2 ESI+: 510, 512
73 PEx2 ESI+: 496, 498
74 PEx2 ESI+: 510, 512
75 PEx2 APCI/ESI+: 460
76 PEx2 ESI+: 588, 590
77 PEx2 APCI/ESI+: 498
78 PEx3 ESI+: 434, 436
79 PEx3 ESI+: 448, 450
80 PEx3 ESI+: 514, 516
81 PEx3 ESI+: 514, 516
82 PEx3 ESI+: 502, 504
83 PEx3 ESI+: 502, 504
84 PEx3 ESI+: 500, 502
85 PEx3 ESI+: 514, 516
86 PEx3 ESI+: 502, 504
87 PEx3 ESI+: 554, 556
88 PEx3 ESI+: 468, 470
89 PEx3 ESI+: 513, 515
90 PEx3 ESI+: 415, 417
91 PEx5 ESI+: 372
92 PEx5 ESI−: 312
93 PEx5 ESI+: 372
94 PEx5 ESI+: 376, 378
95 PEx5 ESI+: 360
96 PEx5 ESI+: 374
97 PEx5 ESI+: 400
98 PEx5 ESI+: 356
99 PEx5 ESI+: 382
100 PEx5 ESI+: 396
TABLE 66
PEx PSyn Data
101 PEx5 ESI+: 356
102 PEx5 ESI+: 382
103 PEx5 NMR-DMSO-d6: 1.11 (3H, d, J = 6 Hz), 1.30-1.41 (1H,
m), 1.59-1.69 (2H, m), 1.87-1.98 (1H, m), 2.05-2.15 (1H,
m), 2.35-2.45 (1H, m), 2.94-3.02 (1H, m), 3.18 (1H, d, J =
14 Hz), 3.97 (3H, d, J = 2 Hz), 3.98 (1H, d, J = 14 Hz),
6.98 (2H, brs), 7.87 (1H, brs), 8.02 (1H, dd, J = 13, 2 Hz)
104 PEx5 NMR-DMSO-d6: 1.14 (3H, d, J = 6 Hz), 1.30-1.42 (1H,
m), 1.58-1.70 (2H, m), 1.87-1.98 (1H, m), 2.04-2.14 (1H,
m), 2.34-2.44 (1H, m), 2.95-3.03 (1H, m), 3.14 (1H, d, J =
14 Hz), 3.91 (3H, d, J = 1 Hz), 3.98 (1H, d, J = 14 Hz),
6.93 (2H, brs), 7.63 (1H, dd, J = 13, 2 Hz), 7.72 (1H, t,
J = 2 Hz)
105 PEx5 NMR-DMSO-d6: 1.08 (3H, d, J = 6 Hz), 1.29-1.41 (1H,
m), 1.58-1.70 (2H, m), 1.86-1.97 (1H, m), 2.05-2.17 (1H,
m), 2.34-2.45 (1H, m), 2.94-3.03 (1H, m), 3.22 (1H, d, J =
14 Hz), 3.96 (1H, d, J = 14 Hz), 6.96 (2H, brs), 7.42 (1H,
t, J = 73 Hz), 7.48 (1H, d, J = 9 Hz), 8.04 (1H, dd, J = 9,
2 Hz), 8.14 (1H, d, J = 2 Hz)
106 PEx5 ESI+: 400
107 PEx5 ESI+: 370
108 PEx5 ESI+: 370
109 PEx5 ESI+: 356
110 PEx5 ESI+: 370
111 PEx5 ESI+: 420, 422
112 PEx5 ESI+: 448, 450
113 PEx6 ESI+: 374
114 PEx6 ESI+: 362
115 PEx6 ESI+: 362
116 PEx7 ESI+: 376, 378
117 PEx7 ESI+: 360
118 PEx7 ESI+: 376
119 PEx7 ESI+: 392
120 PEx7 ESI+: 374
121 PEx7 ESI+: 414
122 PEx9 ESI+: 201
TABLE 67
PEx PSyn Data
123 PEx9 ESI+: 201
124 PEx9 ESI+: 201
125 PEx9 ESI+: 187
126 PEx11 ESI+: 414
127 PEx11 ESI+: 356
128 PEx11 ESI+: 414
129 PEx11 ESI+: 416
130 PEx11 ESI+: 398
131 PEx11 ESI+: 424
132 PEx11 ESI+: 438
133 PEx11 ESI+: 398
134 PEx11 ESI+: 442
135 PEx11 ESI+: 424
136 PEx11 APCI/ESI+: 432
137 PEx11 APCI/ESI+: 398
138 PEx11 ESI+: 450
139 PEx11 ESI+: 442
140 PEx11 ESI+: 412
141 PEx11 ESI+: 412
142 PEx11 ESI+: 398
143 PEx11 ESI+: 412
144 PEx11 ESI+: 462, 464
145 PEx11 ESI+: 490, 492
146 PEx11 ESI+: 416
147 PEx11 ESI+: 416
148 PEx11 ESI+: 404
149 PEx11 ESI+: 404
150 PEx11 ESI+: 402
151 PEx18 ESI+: 632, 634
152 PEx11 ESI+: 418
153 PEx11 ESI+: 434
154 PEx11 ESI+: 416
155 PEx11 ESI+: 456
156 PEx12 ESI+: 418, 420
TABLE 68
PEx PSyn Data
157 PEx13 ESI+: 389
158 PEx13 ESI+: 331
159 PEx13 ESI+: 389
160 PEx13 ESI+: 393
161 PEx13 ESI+: 359
162 PEx13 ESI+: 399
163 PEx13 ESI+: 373
164 PEx13 ESI+: 399
165 PEx13 APCI/ESI+: 407
166 PEx13 APCI/ESI+: 373
167 PEx13 ESI+: 425
168 PEx13 ESI+: 417
169 PEx13 ESI+: 437, 439
170 PEx13 ESI+: 393, 395
171 PEx14 ESI+: 593, 595
172 PEx16 ESI+: 402
173 PEx17 ESI+: 372
174 PEx17 ESI+: 386
175 PEx17 ESI+: 400
176 PEx17 ESI+: 400
177 PEx17 ESI+: 414
178 PEx17 ESI+: 294
179 PEx17 ESI+: 308
180 PEx17 ESI+: 314
181 PEx17 APCI/ESI+: 320
182 PEx17 APCI/ESI+: 348
183 PEx17 APCI/ESI+: 358, 360
184 PEx17 ESI+: 362
185 PEx17 ESI+: 373
186 PEx17 ESI+: 328, 330
187 PEx18 ESI−: 510
188 PEx18 ESI+: 526
189 PEx19 ESI+: 321
190 PEx19 ESI+: 317
TABLE 69
PEx PSyn Data
191 PEx19 ESI+: 317
192 PEx19 ESI+: 259
193 PEx19 ESI+: 321
194 PEx19 ESI+: 317
195 PEx19 ESI+: 305
196 PEx19 ESI+: 287
197 PEx19 ESI+: 327
198 PEx19 ESI+: 301
199 PEx19 ESI+: 327
200 PEx19 ESI+: 335
201 PEx19 ESI+: 301
202 PEx19 ESI+: 353
203 PEx19 ESI+: 345
204 PEx19 ESI+: 365, 367
205 PEx20 NMR-DMSO-d6: 3.87 (3H, s), 8.01 (1H, s), 8.05 (2H, s)
206 PEx20 ESI+: 385
207 PEx20 ESI+: 399
208 PEx20 ESI+: 293
209 PEx20 ESI+: 313, 315
210 PEx20 APCI/ESI+: 319
211 PEx20 APCI/ESI+: 347
212 PEx20 APCI/ESI+: 357
213 PEx20 ESI+: 372
214 PEx21 NMR-DMSO-d6: 1.13-1.26 (9H, m), 1.34-1.45 (1H, m),
1.60-1.76 (2H, m), 1.90-2.12 (2H, m), 2.16-2.28 (2H,
m), 2.45-2.70 (5H, m), 2.78-2.85 (1H, m), 2.92-2.99
(1H, m), 3.00-3.07 (1H, m), 3.10-3.22 (1H, m), 3.55-
3.62 (1H, m), 4.01-4.14 (2H, m), 4.15-4.23 (1H, m),
4.26-4.35 (1H, m), 4.66-4.78 (1H, m), 7.45 (1H, d, J =
1.5 Hz), 7.58 (1H, d, J = 1.3 Hz), 8.32 (1H, d, J = 1.1
Hz), 8.75 (1H, d, J = 1.2 Hz), 11.57 (1H, s)
215 PEx21 ESI−: 616
216 PEx22 ESI+: 604
217 PEx22 ESI+: 604
218 PEx24 ESI+: 541, 543
TABLE 70
PEx PSyn Data
219 PEx24 ESI+: 523
220 PEx29 ESI+: 676
221 PEx29 ESI+: 666, 668
222 PEx29 ESI+: 666, 668
223 PEx29 ESI+: 652, 654
224 PEx29 ESI+: 652, 654
225 PEx29 ESI+: 666, 668
226 PEx29 ESI+: 664
227 PEx29 ESI+: 666, 668
228 PEx29 ESI+: 664
229 PEx29 ESI+: 690
230 PEx29 ESI+: 618, 620
231 PEx29 ESI+: 618
232 PEx29 ESI+: 664
233 PEx29 ESI+: 690
234 PEx29 ESI+: 618
235 PEx29 ESI+: 618
236 PEx29 ESI+: 710, 712
237 PEx29 ESI+: 632, 634
238 PEx29 ESI+: 632, 634
239 PEx29 ESI+: 738, 740
240 PEx29 ESI+: 752, 754
241 PEx29 ESI+: 752, 754
242 PEx29 ESI+: 752, 754
243 PEx29 ESI+: 738, 740
244 PEx29 ESI+: 724, 726
245 PEx29 ESI+: 724, 726
246 PEx29 ESI−: 519, 521
247 PEx29 ESI+: 664
248 PEx29 ESI+: 650
249 PEx29 ESI+: 663
250 PEx30 ESI+: 245
251 PEx30 ESI+: 279
252 PEx30 ESI+: 279, 281
TABLE 71
PEx PSyn Data
253 PEx30 ESI+: 263
254 PEx30 ESI+: 275
255 PEx30 ESI+: 275
256 PEx30 ESI+: 303
257 PEx30 ESI+: 303
258 PEx30 ESI+: 259, 261
259 PEx30 ESI+: 275
260 PEx30 ESI+: 285
261 PEx30 ESI+: 293
262 PEx30 ESI+: 275
263 PEx30 ESI+: 289
264 PEx30 ESI+: 285
265 PEx30 ESI+: 259
266 PEx30 ESI+: 323, 325
267 PEx30 ESI+: 197
268 PEx30 ESI+: 217, 219
269 PEx30 APCI/ESI+: 223
270 PEx30 APCI/ESI+: 251
271 PEx30 ESI+: 211
272 PEx30 ESI+: 233
273 PEx30 ESI+: 251, 253
274 PEx30 APCI/ESI+: 261, 263
275 PEx30 ESI+: 276
276 PEx38 EI: 228
277 PEx40 ESI+: 155
278 PEx40 EI: 194, 196
279 PEx41 EI: 202
280 PEx43 ESI+: 604
281 PEx43 ESI+: 604
282 PEx43 ESI+: 618
283 PEx45 ESI+: 301
284 PEx45 ESI+: 301
285 PEx45 ESI+: 301
TABLE 72
Ex Structure
1
Figure USRE049111-20220621-C00294
2
Figure USRE049111-20220621-C00295
3
Figure USRE049111-20220621-C00296
4
Figure USRE049111-20220621-C00297
5
Figure USRE049111-20220621-C00298
TABLE 73
Ex Structure
6
Figure USRE049111-20220621-C00299
7
Figure USRE049111-20220621-C00300
8
Figure USRE049111-20220621-C00301
9
Figure USRE049111-20220621-C00302
TABLE 74
Ex Structure
10
Figure USRE049111-20220621-C00303
11
Figure USRE049111-20220621-C00304
12
Figure USRE049111-20220621-C00305
13
Figure USRE049111-20220621-C00306
TABLE 75
Ex Structure
14
Figure USRE049111-20220621-C00307
15
Figure USRE049111-20220621-C00308
16
Figure USRE049111-20220621-C00309
17
Figure USRE049111-20220621-C00310
TABLE 76
Ex Structure
18
Figure USRE049111-20220621-C00311
19
Figure USRE049111-20220621-C00312
20
Figure USRE049111-20220621-C00313
21
Figure USRE049111-20220621-C00314
22
Figure USRE049111-20220621-C00315
TABLE 77
Ex Structure
23
Figure USRE049111-20220621-C00316
24
Figure USRE049111-20220621-C00317
25
Figure USRE049111-20220621-C00318
26
Figure USRE049111-20220621-C00319
27
Figure USRE049111-20220621-C00320
TABLE 78
Ex Structure
28
Figure USRE049111-20220621-C00321
29
Figure USRE049111-20220621-C00322
30
Figure USRE049111-20220621-C00323
31
Figure USRE049111-20220621-C00324
32
Figure USRE049111-20220621-C00325
TABLE 79
Ex Structure
33
Figure USRE049111-20220621-C00326
34
Figure USRE049111-20220621-C00327
35
Figure USRE049111-20220621-C00328
36
Figure USRE049111-20220621-C00329
TABLE 80
Ex Structure
37
Figure USRE049111-20220621-C00330
38
Figure USRE049111-20220621-C00331
39
Figure USRE049111-20220621-C00332
40
Figure USRE049111-20220621-C00333
TABLE 81
Ex Structure
41
Figure USRE049111-20220621-C00334
42
Figure USRE049111-20220621-C00335
43
Figure USRE049111-20220621-C00336
44
Figure USRE049111-20220621-C00337
TABLE 82
Ex Structure
45
Figure USRE049111-20220621-C00338
46
Figure USRE049111-20220621-C00339
47
Figure USRE049111-20220621-C00340
48
Figure USRE049111-20220621-C00341
TABLE 83
Ex Structure
49
Figure USRE049111-20220621-C00342
50
Figure USRE049111-20220621-C00343
51
Figure USRE049111-20220621-C00344
52
Figure USRE049111-20220621-C00345
TABLE 84
Ex Structure
53
Figure USRE049111-20220621-C00346
54
Figure USRE049111-20220621-C00347
55
Figure USRE049111-20220621-C00348
56
Figure USRE049111-20220621-C00349
TABLE 85
Ex Structure
57
Figure USRE049111-20220621-C00350
58
Figure USRE049111-20220621-C00351
59
Figure USRE049111-20220621-C00352
60
Figure USRE049111-20220621-C00353
TABLE 86
Ex Structure
61
Figure USRE049111-20220621-C00354
62
Figure USRE049111-20220621-C00355
63
Figure USRE049111-20220621-C00356
64
Figure USRE049111-20220621-C00357
TABLE 87
Ex Structure
65
Figure USRE049111-20220621-C00358
66
Figure USRE049111-20220621-C00359
67
Figure USRE049111-20220621-C00360
68
Figure USRE049111-20220621-C00361
TABLE 88
Ex Structure
69
Figure USRE049111-20220621-C00362
70
Figure USRE049111-20220621-C00363
71
Figure USRE049111-20220621-C00364
72
Figure USRE049111-20220621-C00365
TABLE 89
Ex Structure
73
Figure USRE049111-20220621-C00366
74
Figure USRE049111-20220621-C00367
75
Figure USRE049111-20220621-C00368
76
Figure USRE049111-20220621-C00369
77
Figure USRE049111-20220621-C00370
TABLE 90
Ex Structure
78
Figure USRE049111-20220621-C00371
79
Figure USRE049111-20220621-C00372
80
Figure USRE049111-20220621-C00373
81
Figure USRE049111-20220621-C00374
82
Figure USRE049111-20220621-C00375
TABLE 91
Ex Structure
83
Figure USRE049111-20220621-C00376
84
Figure USRE049111-20220621-C00377
85
Figure USRE049111-20220621-C00378
86
Figure USRE049111-20220621-C00379
87
Figure USRE049111-20220621-C00380
TABLE 92
Ex Structure
88
Figure USRE049111-20220621-C00381
89
Figure USRE049111-20220621-C00382
90
Figure USRE049111-20220621-C00383
91
Figure USRE049111-20220621-C00384
92
Figure USRE049111-20220621-C00385
TABLE 93
Ex Structure
93
Figure USRE049111-20220621-C00386
94
Figure USRE049111-20220621-C00387
95
Figure USRE049111-20220621-C00388
96
Figure USRE049111-20220621-C00389
97
Figure USRE049111-20220621-C00390
TABLE 94
Ex Structure
98
Figure USRE049111-20220621-C00391
99
Figure USRE049111-20220621-C00392
100
Figure USRE049111-20220621-C00393
101
Figure USRE049111-20220621-C00394
102
Figure USRE049111-20220621-C00395
TABLE 95
Ex Structure
103
Figure USRE049111-20220621-C00396
104
Figure USRE049111-20220621-C00397
105
Figure USRE049111-20220621-C00398
106
Figure USRE049111-20220621-C00399
TABLE 96
Ex Structure
107
Figure USRE049111-20220621-C00400
108
Figure USRE049111-20220621-C00401
109
Figure USRE049111-20220621-C00402
110
Figure USRE049111-20220621-C00403
111
Figure USRE049111-20220621-C00404
TABLE 97
Ex Structure
112
Figure USRE049111-20220621-C00405
113
Figure USRE049111-20220621-C00406
114
Figure USRE049111-20220621-C00407
115
Figure USRE049111-20220621-C00408
TABLE 98
Ex Structure
116
Figure USRE049111-20220621-C00409
117
Figure USRE049111-20220621-C00410
118
Figure USRE049111-20220621-C00411
119
Figure USRE049111-20220621-C00412
TABLE 99
Ex Structure
120
Figure USRE049111-20220621-C00413
121
Figure USRE049111-20220621-C00414
122
Figure USRE049111-20220621-C00415
123
Figure USRE049111-20220621-C00416
124
Figure USRE049111-20220621-C00417
TABLE 100
Ex Structure
125
Figure USRE049111-20220621-C00418
126
Figure USRE049111-20220621-C00419
127
Figure USRE049111-20220621-C00420
128
Figure USRE049111-20220621-C00421
129
Figure USRE049111-20220621-C00422
130
Figure USRE049111-20220621-C00423
TABLE 101
Ex Structure
131
Figure USRE049111-20220621-C00424
132
Figure USRE049111-20220621-C00425
133
Figure USRE049111-20220621-C00426
134
Figure USRE049111-20220621-C00427
TABLE 102
Ex Structure
135
Figure USRE049111-20220621-C00428
136
Figure USRE049111-20220621-C00429
137
Figure USRE049111-20220621-C00430
138
Figure USRE049111-20220621-C00431
TABLE 103
Ex Structure
139
Figure USRE049111-20220621-C00432
140
Figure USRE049111-20220621-C00433
141
Figure USRE049111-20220621-C00434
142
Figure USRE049111-20220621-C00435
TABLE 104
Ex Structure
143
Figure USRE049111-20220621-C00436
144
Figure USRE049111-20220621-C00437
TABLE 105
Ex. Syn. Data
1 Ex1 ESI+: 592, 592
NMR-DMSO-d6: 1.20-1.52 (6H, m), 1.60-1.81 (1H, m), 1.85-
2.03 (2H, m), 2.15-2.29 (1H, m), 2.77-300 (2H, m), 3.09-
3.75 (9H, m), 3.75-4.77 (5H, m), 4.84-4.97 (1H, m), 7.65-
7.71 (1H, m), 7.74 (1H, d, J = 1.3 Hz), 8.48-8.56 (1H, m),
8.79-8.85 (1H, m), 10.60-11.20 (1H, m), 11.45-11.84 (1H,
m), 12.20-12.38 (1H, m)
2 Ex2 ESI+: 657
3 Ex3 ESI+: 624
NMR-DMSO-d6, 1.40-1.54 (6H, m), 1.61-1.75 (1H, m), 1.84-
2.04 (2H, m), 2.16-2.28 (1H, m), 2.84-3.12 (3H, m), 3.16-
3.30 (3H, m), 3.46-3.62 (6H, m), 4.55-4.72 (2H, m), 4.94
(1H, d, J = 15 Hz), 5.05 (1H, brs), 7.95 (1H, s), 8.40 (1H, t,
J = 1 Hz), 8.47 (1H, s), 8.85 (1H, d, J = 1 Hz), 10.6 (1H,
brs), 11.1 (1H, brs), 12.4 (1H, s), 12.7 (1H, brs)
4 Ex4 ESI+: 632
5 Ex5 ESI+: 548, 550 (M − H)−
6 Ex6 ESI+: 570
7 Ex7 ESI+: 662
8 Ex8 ESI+: 648
9 Ex9 ESI+: 624
NMR-DMSO-d6, 1.05-1.15 (6H, m), 1.20-1.52 (3H, m), 2.75-
4.20 (14H, m), 4.40-4.80 (4H, m), 7.80-7.87 (1H, m), 7.88-
7.96 (2H, m), 8.53 (1H, s), 8.84 (1H, s), 10.63 (1H, brs),
11.33-11.76 (1H, m), 12.30-12.42 (1H, m)
10 Ex1 ESI+: 648
11 Ex1 ESI+: 638, 640
12 Ex1 ESI+: 638, 640
13 Ex1 ESI+: 624
14 Ex1 ESI+: 624
15 Ex1 ESI+: 638
16 Ex1 ESI+: 638, 640
17 Ex1 ESI+: 636
18 Ex1 ESI+: 663
19 Ex1 ESI+: 590
20 Ex1 ESI+: 590, 592
21 Ex1 ESI+: 636
TABLE 106
Ex. Syn. Data
22 Ex1 ESI+: 576, 578
NMR-DMSO-d6; 1.34-1.50 (6H, m), 1.62-1.73 (1H, m), 1.84-
2.02 (2H, m), 2.17-2.28 (1H, m), 2.92-4.27 (13H, m), 4.55-
4.69 (2H, m), 4.87-4.96 (1H, m), 5.04 (1H, brs), 7.68 (1H, d,
J = 1.4 Hz), 7.74 (1H, d, J = 1.4 Hz), 8.43 (1H, s), 8.84 (1H,
d, J = 1.2 Hz), 10.57 (1H, brs), 12.32 (1H, s)
23 Ex1 ESI+: 662
24 Ex1 ESI+: 576, 578
25 Ex1 ESI+: 590
26 Ex1 ESI+: 576
27 Ex1 ESI+: 682, 684
28 Ex1 ESI+: 604
NMR-DMSO-d6; 0.89 (3H, t, J = 7.3 Hz), 1.47 (3H, d, J =
7.1 Hz), 1.60-1.74 (2H, m), 1.83-2.01 (3H, m), 2.16-2.26
(1H, m), 2.85-3.01 (14H, m), 4.58-4.77 (2H, m), 4.87-4.96
(1H, m), 5.02-5.11 (1H, m), 7.70 (1H, d, J = 1.3 Hz), 7.74
(1H, d, J = 1.3 Hz), 8.47 (1H, s), 8.84 (1H, d, J = 1.1 Hz),
10.59 (1H, brs), 11.11 (1H, brs), 12.32 (1H, s)
29 Ex1 ESI+: 604
28 Ex1 ESI+: 636
NMR-DMSO-d6; 1.36 (3H, t, J = 6.4 Hz), 1.44 (3H, d, J =
7.1 Hz), 1.59-1.69 (1H, m), 1.85-1.96 (2H, m), 2.14-2.22
(1H, m), 2.87-3.20 (4H, m), 3.20-3.74 (9H, m), 4.47-4.53
(1H, m), 4.61-4.69 (1H, m), 4.79-4.85 (1H, m), 5.03-5.10
(1H, m), 7.80-7.85 (1H, m), 7.91-7.96 (2H, m), 8.47 (1H, s),
8.86 (1H, d, J = 1.2 Hz), 10.48 (1H, brs), 10.76 (1H, brs)
12.34-12.38 (1H, m)
31 Ex1 ESI+: 604
NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.18-1.52 (3H, m),
1.58-1.75 (2H, m), 1.81-2.02 (3H, m), 2.14-2.27 (1H, m),
2.78-2.95 (2H, m), 3.07-3.98 (10H, m), 4.37-4.78 (3H, m),
4.85-4.98 (1H, m), 7.69 (1H, s), 7.74 (1H, d, J = 1.2 Hz),
8.52 (1H, s), 8.80-8.84 (1H, m), 10.59 (1H, brs), 11.29-11.79
(1H, m), 12.32 (3H, s), 12.50-13.07 (1H, m)
TABLE 107
Ex. Syn. Data
32 Ex1 ESI+: 622
NMR-DMSO-d6; 1.14-1.51 (6H, m), 1.61-1.78 (1H, m), 1.83-
2.00 (2H, m), 2.11-2.24 (1H, m), 3.00-4.30 (12H, m), 4.48
(1H, dd, J = 7.4, 14.8 Hz), 4.64 (1H, d, J = 14.0 Hz), 4.76
(1H, d, J = 14.5 Hz), 5.07 (1H, brs), 7.77-7.83 (1H, m), 7.90-
8.04 (2H, m), 8.41-8.48 (1H, m), 8.85 (1H, d, J = 1.3 Hz),
11.05-11.60 (1H, m), 12.28-12.42 (1H, m)
33 Ex1 ESI+: 635
NMR-DMSO-d6; 1.36 (3H, d, J = 6.3 Hz), 1.59-1.69 (1H,
m), 1.83-1.97 (2H, m), 2.13-2.22 (1H, m), 2.85-2.89 (2H, m),
3.03-3.78 (11H, m), 4.67 (3H, s), 4.39-4.87 (4H, m), 8.38-
8.41 (1H, m), 8.49-8.52 (1H, m), 8.75-8.80 (1H, m), 8.84
(1H, d, J = 1.3 Hz), 10.50 (1H, brs), 10.76-11.73 (1H, m),
12.34 (1H, s), 12.40-12.90 (1H, br)
34 Ex2 ESI+: 629
35 Ex2 ESI+: 671
36 Ex2 ESI+: 671
37 Ex2 ESI+: 671
38 Ex2 ESI+: 657
39 Ex2 ESI+: 643
40 Ex2 ESI+: 643
41 Ex3 ESI+: 634
42 Ex3 ESI+: 646 [M − H]−
43 Ex3 ESI+: 634, 636
44 Ex3 ESI+: 634
45 Ex3 ESI+: 620, 622
46 Ex3 ESI+: 634
47 Ex3 ESI+: 620
48 Ex3 ESI+: 634
49 Ex3 ESI+: 622
NMR-DMSO-d6; 1.34-1.40 (3H, m), 1.58-1.76 (1H, m), 1.83-
1.97 (2H, m), 2.11-2.23 (1H, m), 2.90 (2H, t, J = 7.6 Hz),
3.03-3.97 (13H, m), 4.42-4.56 (1H, m), 4.60-4.87 (3H, m),
7.79-7.85 (1H, m), 7.91-8.00 (2H, m), 8.51 (1H, d, J =
1.2 Hz), 8.85 (1H, d, J = 1.3 Hz), 10.75 (1H, brs), 11.05-
11.45 (1H, m), 12.33-12.41 (1H, m)
TABLE 108
Ex. Syn. Data
50 Ex3 ESI+: 576, 578
NMR-DMSO-d6; 1.44 (3H, t, J = 6.5 Hz), 1.62-1.73 (1H,
m), 1.84-2.02 (2H, m), 2.17-2.28 (1H, m), 2.90 (2H, t, J =
7.7 Hz), 3.08-3.23 (3H, m), 3.30-3.39 (2H, m), 3.42-4.07
(8H, m), 4.56-4.77 (3H, m), 4.88-4.96 (1H, m), 7.68 (1H, d,
J = 1.4 Hz), 7.74 (1H, d, J = 1.4 Hz), 8.51 (1H, d, J =
1.2 Hz), 8.83 (1H, d, J = 1.2 Hz), 10.45-11.00 (1H, m)
51 Ex3 ESI+: 562
52 Ex9 ESI+: 634
53 Ex3 ESI+: 620
54 Ex3 ESI+: 648
55 Ex3 ESI+: 662
56 Ex3 ESI+: 662
57 Ex3 ESI+: 662
58 Ex3 ESI+: 676
59 Ex3 ESI+: 676
60 Ex3 ESI+: 622
61 Ex3 ESI+: 608
62 Ex3 ESI+: 622
63 Ex3 ESI+: 644
64 Ex3 ESI+: 622
65 Ex3 ESI+: 618
66 Ex3 ESI+: 604
67 Ex3 ESI+: 664
68 Ex3 ESI+: 630
69 Ex3 ESI+: 658
70 Ex3 ESI+: 644
71 Ex3 ESI+: 604
72 Ex3 ESI+: 652
73 Ex3 ESI+: 618
74 Ex3 ESI+: 632, 634
75 Ex3 ESI+: 662
76 Ex3 ESI+: 670
77 Ex3 ESI+: 556
78 Ex3 ESI+: 570
TABLE 109
Ex. Syn. Data
79 Ex3 ESI+: 590
80 Ex3 ESI+: 590, 592
NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.58-1.72 (2H,
m), 1.82-2.02 (3H, m), 2.16-2.27 (1H, m), 2.89 (2H, t, J =
7.6 Hz), 3.06-3.70 (13H, m), 4.60-4.78 (3H, m), 4.89-4.98
(1H, m), 7.69 (1H, d, J = 1.3 Hz), 7.74 (1H, d, J = 1.4 Hz),
8.51 (1H, d, J = 1.2 Hz), 8.84 (1H, d, J = 1.3 Hz), 10.43
(1H, brs), 11.30 (1H, brs), 12.35 (1H, s)
81 Ex3 ESI+: 604, 606
82 Ex9 ESI+: 576
83 Ex3 ESI+: 562
84 Ex3 ESI+: 590
85 Ex3 ESI+: 568
86 Ex3 ESI+: 582
87 Ex3 ESI+: 596
88 Ex3 ESI+: 596
89 Ex3 ESI+: 596
90 Ex3 ESI+: 596
91 Ex3 ESI+: 666
92 Ex3 ESI+: 666
93 Ex3 ESI+: 638
94 Ex3 ESI+: 610
95 Ex3 ESI+: 624
96 Ex3 ESI+: 624
97 Ex3 ESI+: 620, 622
98 Ex3 ESI+: 636, 638
99 Ex3 ESI+: 634, 636
100 Ex3 ESI+: 624
101 Ex3 ESI+: 634
102 Ex3 ESI+: 634
103 Ex3 ESI+: 596
104 Ex3 ESI+: 650
105 Ex3 ESI+: 650
106 Ex3 ESI+: 624
TABLE 110
Ex. Syn. Data
107 Ex3 ESI+: 624
NMR-DMSO-d6; 0.85 (3H, d, J = 6.6 Hz), 0.90 (3H, d, J =
6.5 Hz), 1.90-2.00 (1H, m), 2.60-2.71 (3H, m), 2.71-2.81
(2H, m), 2.91 (2H, t, J = 7.7 Hz), 3.05-3.25 (2H, m), 3.28-
3.38 (2H, m), 3.49-3.67 (4H, m), 3.80-4.82 (5H, m), 7.82-
7.90 (3H, m), 8.51 (1H, d, J = 1.2 Hz), 8.85 (1H , d, J = 1.3
Hz), 10.25 (1H, brs), 11.62 (1H, brs), 12.36 (1H, s)
108 Ex3 ESI+: 622
109 Ex9 ESI+: 610
110 Ex3 ESI+: 636
111 Ex3 ESI+: 624
NMR-DMSO-d6; 1.27 (3H, d, J = 6.8 Hz), 1.42 (3H, d, J =
6.5 Hz), 1.55-1.80 (2H, m), 2.12-2.34 (2H, m), 2.92 (2H, t,
J = 7.7 Hz), 3.05-3.25 (2H, m), 3.25-3.40 (2H, m), 3.48-
3.79 (5H, m), 3.82-3.97 (1H, m), 4.51 (1H, dd, J = 7.2,
15.3 Hz), 4.59-4.83 (3H, m), 4.90-6.65 (2H, m), 7.96-7.99
(1H, m), 8.39-8.42 (1H, m), 8.49-8.53 (1H, m), 8.54 (1H,
d, J = 1.3 Hz), 10.99 (1H, brs), 11.75 (1H, brs), 12.37-
(1H, s)
112 Ex3 ESI+: 676
113 Ex3 ESI+: 690
114 Ex3 ESI+: 588, 590 [M − H]−
115 Ex3 ESI+: 602, 604 [M − H]−
116 Ex3 ESI+: 602, 604 [M − H]−
117 Ex3 ESI+: 602, 604 [M − H]−
118 Ex9 ESI+: 610
NMR-DMSO-d6; 1.06-1.14 (6H, m), 1.43 (3H, d, J =
6.8 Hz), 2.92-3.15 (4H, m), 3.16-3.45 (2H, m), 3.51-3.75
(4H, m), 3.80-4.84 (6H, m), 4.97-5.24 (1H, m), 7.80-7.86
(1H, m), 7.87-7.97 (2H, m), 8.42-8.48 (1H, m), 8.86 (1H,
d, J = 1.2 Hz), 10.06-11.50 (2H, m), 12.37 (1H, s)
119 Ex3 ESI+: 636
NMR-DMSO-d6; 1.21 (3H, d, J = 6.8 Hz), 1.40 (3H, d, J =
6.5 Hz), 1.60-1.71 (2H, m), 2.17-2.30 (2H, m), 2.91 (2H, t,
J = 7.7 Hz), 3.08-3.25 (2H, m), 3.29-3.37 (2H, m), 3.50-
3.70 (4H, m), 3.87-3.98 (1H, m), 4.38 (1H, dd, J = 7.5,
15.3 Hz), 4.50-5.00 (5H, m), 7.79-7.85 (1H, m), 7.94-8.00
(2H, m), 8.51 (1H, d, J = 1.2 Hz), 8.85 (1H, d, J =
1.3 Hz), 11.08 (1H, m), 11.63 (1H, brs), 12.37 (1H, s)
TABLE 111
Ex. Syn. Data
120 Ex3 ESI+: 622
NMR-DMSO-d6; 1.22 (3H, d, J = 6.8 Hz), 1.41 (3H, d, J =
6.5 Hz), 1.57-1.75 (2H, m), 2.12-2.33 (2H, m), 3.11-4.03
(7H, m), 4.26 (2H, s), 4.25-5.21 (7H, m), 7.78-7.83 (1H,
m), 7.95-8.03 (2H, m), 8.50 (1H, d, J = 1.2 Hz), 8.85
(1H, d, J = 1.3 Hz), 10.60-11.70 (2H, m), 12.38 (1H, s)
121 Ex3 ESI+: 650
122 Ex3 ESI+: 632
123 Ex4 ESI+: 604
124 Ex4 ESI+: 632
125 Ex5 ESI−: 576, 578 [M − H]−
126 Ex5 ESI−: 602, 604 [M − H]−
127 Ex5 ESI−: 576, 578 [M − H]−
128 Ex5 ESI−: 588, 590 [M − H]−
129 Ex5 ESI−: 576, 578 [M − H]−
130 Ex5 ESI−: 588, 590 [M − H]−
NMR-DMSO-d6; 0.33-0.54 (2H, m), 0.60-0.72 (2H, m),
1.13-1.50 (4H, m), 2.75-2.81 (3H, m), 2.81-3.07 (3H, m),
3.11-3.74 (7H, m), 3.74-4.94 (7H, m), 7.68 (1H, d, J =
1.4 Hz), 7.74 (1H, d, J = 1.4 Hz), 8.47-8.57 (1H, m),
8.78-8.86 (1H, m), 10.58 (1H, brs), 11.33-11.77 (1H, m),
12.22-12.42 (1H, m)
131 Ex6 ESI+: 610, 612
132 Ex6 ESI+: 592
133 Ex7 ESI+: 676
134 Ex7 ESI+: 676
135 Ex7 ESI+: 690
136 Ex8 ESI+: 662
137 Ex8 ESI+: 676
138 Ex9 ESI+: 638
139 Ex9 ESI+: 654
140 Ex1 ESI+: 636 [M + H]+
NMR-DMSO-d6; 1.34-1.41 (3H, m), 1.47 (3H, d, J =
7.0 Hz), 1.60-1.74 (1H, m), 1.86-1.97 (2H, m), 2.11-2.23
(1H, m), 2.81-4.17 (15H, m), 4.43-4.54 (1H, m), 4.61-4.71
(1H, m), 4.75-4.84 (1H, m), 5.01-5.12 (1H, m), 7.79-7.84
(1H, m), 7.91-8.00 (2H, m), 8.45-8.49 (1H, m), 8.85 (1H,
d, J = 1.2 Hz), 10.85 (1H, brs), 10.95-11.31 (1H,
m), 12.30-12.41 (1H, m)
TABLE 112
Ex. Syn. Data
141 Ex1 ESI+: 622 [M + H]+
NMR-DMSO-d6; 1.34-1.47 (6H, m), 1.60-1.74 (1H, m),
1.86-1.97 (2H, m), 2.11-2.23 (1H, m), 3.10-4.29
(13H, m), 4.42-4.54 (1H, m), 4.57-4.72 (1H, m), 4.74-
4.83 (1H, m), 5.09 (1H, brs), 7.78-7.85 (1H, m), 7.91-
8.02 (2H, m), 8.45 (1H, s), 8.86 (1H, d, J = 1.3 Hz),
10.75-11.42 (1H, m), 12.32-12.41 (1H, m)
142 Ex3 ESI+: 590, 592 [M + H]+
NMR-DMSO-d6; 1.21-1.31 (1H, m), 1.35-1.49 (6H, m),
1.61-1.74 (1H, m), 1.84-2.02 (2H, m), 2.13-2.29
(1H, m), 2.79-2.98 (2H, m), 3.10-3.98 (10H, m),
4.40-4.77 (3H, m), 4.87-4.98 (1H, m), 7.67 (1H, d, J =
1.3 Hz), 7.74 (1H, d, J = 1.3 Hz), 8.52 (1H, s), 8.82
(1H, s), 10.39-10.97 (1H, m), 11.25-11.65 (1H, m),
12.25-12.42 (1H, m), 12.56-13.02 (1H, br)
143 Ex3 ESI+: 604, 606 [M + H]+
NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.22-1.30
(1H, m), 1.40-1.49 (2H, m), 1.59-1.73 (2H, m), 1.83-
2.03 (3H, m), 2.14-2.27 (1H, m), 2.80-2.97 (2H, m),
3.10-3.95 (12H, m), 4.42-4.79 (3H, m), 4.88-4.97 (1H,
m), 7.69 (1H, d, J = 1.3 Hz), 7.74 (1H, d, J = 1.3 Hz),
8.52 (1H, s), 8.83 (1H, s), 10.38-11.04 (1H, m), 11.20-
11.24 (1H, m), 12.21-13.19 (2H, m)
144 Ex144 ESI+: 590, 592 [M + H]+
NMR-DMSO-d6; 1.13-1.45 (6H, m), 1.46-2.29 (4H, m),
2.53-2.70 (2H, m), 2.80-3.84 (14H, m), 4.00-5.18
(4H, m), 6.09 (4H, s), 7.57 (1H, s), 7.71 (1H, s),
8.47 (1H, s), 8.79 (1H, d, J = 1.2 Hz), 9.15-10.50
(1H, m), 11.70-12.50 (1H, m)
INDUSTRIAL APPLICABILITY
The compound of the formula (I) or a salt thereof is a muscarinic M3 receptor-positive allosteric modulator, and can thus be used as an agent for preventing or treating bladder/urinary tract diseases associated wit bladder contractions via a muscarinic M3 receptor.

Claims (23)

The invention claimed is:
1. A compound of formula (I) or a salt thereof:
Figure USRE049111-20220621-C00438
wherein
R1 is —N(—R12)(—R12) —N(—R11)(—R12), or optionally-substituted cyclic amino,
R11 is C1-6 alkyl,
R12 is optionally-substituted C1-6 alkyl, or optionally-substituted C3-8 cycloalkyl,
R2 is optionally-substituted aryl, optionally-substituted monocyclic aromatic hetero ring, or optionally-substituted bicyclic aromatic hetero ring,
each R3 if present is, independently, C1-6 alkyl,
W is C1-6 alkylene, and
n is an integer of 0 to 4.
2. The compound or salt thereof according to claim 1, wherein
R1 is cyclic amino optionally substituted with 1 to 5 of a substituent G and/or an oxo substituent, or R3 is —N(—R11)(—R12),
R11 is C1-6 alkyl
R12 is C1-6 alkyl optionally substituted with 1 to 3 substituents selected from the group consisting of
—OH,
 O—C1-6 alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alky), —CN, —SO2—(C1-6 alkyl), and halogen,
C3-8 cycloalkyl,
O—(C3-8 cycloalkyl),
halogen,
—CN, and
a saturated hetero ring,
R2 is phenyl optionally substituted with 1 to 5 substituents G, thienyl optionally substituted with 1 to 3 substituents G, pyridyl optionally substituted with 1 to 3 substituents G, or benzothienyl optionally substituted with 1 to 5 substituents G, and
each substituent G is a sustituent selected from the group consisting of:
C1-6 alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alkyl), —CN, —SO2—(C1-6 alkyl), and halogen,
—OH,
—O—C1-6 alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C1-6 alkyl), —CN, —SO2—(C1-6 alkyl), and halogen,
C3-8 cycloalkyl,
—O—(C3-8 cycloalkyl),
halogen,
—CN,
—SO2—(C1-6 alkyl),
—CO2—(C1-6 alkyl),
—COOH,
—CO—N(C1-6 alkyl)2,
—CO—NH(C1-6 alkyl),
—CONH2,
—CO—(C1-6 alkyl),
—SO2—N(C1-6 alkyl)2,
—SO2—NH(C1-6 alkyl),
—SO2NH2,
—N(C1-6 alkyl)2,
—NH(C1-6 alkykl),
—NH2,
a saturated hetero ring, and
—O-saturated hetero ring.
3. The compound or a salt thereof according to claim 2, wherein
R1 is pyrrolidin-1-yl or piperidin-1-yl, each substituted with 1 to 2 substituents selected from the group consisting of C1-6 alkyl and haogeno-C1-6 alkyl, or wherein R1 is —N(—R11)(—R12),
R11 is C1-6 alkyl, and
R12 is C1-6 alkyl optionally substituted with one group selected from the group consisting of C3-8 cycloalkyl and —O—(C1-6 alkyl),
R2
phenyl optionally substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), —O-(halogeno-C1-6 alkyl), halogen, C3-8 cycloalkyl, and —CN;
thienyl optionally substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C1-6 alkyl), C3-8 cycloalkyl, and halogen;
pyridyl optionally substituted with 1 to 3 groups selected from the group consisting of C1-6 alkyl, halogeno-C1-6 alkyl, —O—(C3-8 alkyl), C3-8 cycloalkyl, and halogen; or
benzothienyl,
W is C1-6 alkylene, and
n is 0 or 1.
4. The compound or a salt thereof according to claim 3, wherein
R2 is phenyl di-substituted with trifluoromethyl and fluoro, thienyl mono-substituted with trifluoromethyl or chloro, or pyridyl di-substituted with trifluoromethyl and methoxy, and
W is methylene or ethylene.
5. The compound or a salt thereof according to claim 3, wherein
R1 is pyrrolidin-1-yl or piperidin-1-yl, each substituted with 1 to 2 substituents selected from the group consisting of C1-6 alkyl and halogeno-C1-6 alkyl,
R2 is thienyl optionally substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen, or wherein R2 is phenyl optionally substituted with 1 or 2 substituents selected from the group consisting of halogeno-C1-6 alkyl and halogen, and
W is methylene or ethylene.
6. The compound or a salt thereof according to claim 1, wherein the compound is a compound selected from the group consisting of:
3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid,
3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid,
[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid,
3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid,
3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid,
3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl)}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid,
3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid, and
3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid.
7. A pharmaceutical composition, comprising:
the compound or a salt thereof according to claim 1; and
a pharmaceutically acceptable excipient.
8. A method for treating a bladder/urinary tract disease associated with bladder contractions via a muscarinic M3 receptor, the method comprising:
administering, to a subject in need thereof, an effective amount of the compound or a salt thereof according to claim 1.
9. The method according to claim 8, wherein the bladder/urinary tract disease associated with bladder contractions via a muscarinic M3 receptor is voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, or neurogenic bladder.
10. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid.
11. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid.
12. The compound or a salt thereof according to claim 6, wherein the compound is [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid.
13. The compound or a salt thereof according to claim 6, wherein the compound is 3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid.
14. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid.
15. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl]-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid.
16. The compound or a salt thereof according to claim 6, wherein the compound is 3-(4-{5-[(5-([(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid.
17. The compound or a salt thereof according to claim 6, wherein the compound is 3-{(2R)-4-[5-([5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid.
18. The compound of claim 1 which is 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid dimaleate.
19. A crystal polymorph of the compound of claim 18.
20. The crystal polymorph of claim 19 having peaks at 2θ (°) of 5.7, 6.6, 10.5, 12.0, 13.3, 15.8, 16.6, 17.3, 19.0, and 26.2 when measured by powder X-ray diffraction.
21. A pharmaceutical composition comprising the crystal polymorph of claim 20.
22. A method for treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, comprising administering the composition of claim 21.
23. A method for treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor voiding dysfunction or urine storage
dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, or neurogenic bladder comprising administering the composition of claim 21.
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