Patent Application
20230312542
The present invention relates to a target protein degradation-inducing compound that is a bifunctional compound having a portion that binds to Von-Hippel-Lindau, which is a substrate recognition protein of a ubiquitin ligase complex constituted of low-molecular-weight compounds (hereinafter sometimes to be referred to as VHL binding ligand), at one end, and a portion that is capable of binding or binds to a target protein (hereinafter sometimes to be referred to as target-directed ligand) at the other end, and is useful in the pharmaceutical field.
The ubiquitin-proteasome system is a system that degrades and removes proteins no longer needed in cells, through ubiquitination of target proteins by proteasomes in an ATP-dependent manner. The system plays an important role in cell proliferation, survival, maintenance of homeostasis, and the like. The ubiquitin-proteasome system involves multiple enzymes called ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), and ubiquitin ligase (E3). The ubiquitination of the target protein is achieved by sequential functions of these enzymes. In particular, a target protein that has undergone so-called polyubiquitination, in which a ubiquitin chain is linked via the 48th lysine residue of ubiquitin, is efficiently recognized and degraded by the proteasome. It has been reported that humans have two types of ubiquitin activating enzymes, about 40 types of ubiquitin conjugating enzymes, and about 600 types of ubiquitin ligases. It is known that substrate recognition by ubiquitin ligases is strictly regulated. Ubiquitin ligases include those that function alone and those that form a complex to function. Particularly in the latter, a complex structure in which a substrate recognition protein binds to a scaffold protein via an adapter protein is taken. This substrate recognition protein determines the substrate protein to be ubiquitinated. In recent years, it has been reported that degradation of proteins that are not originally substrates for ubiquitin ligase can be induced by chemically linking a compound that binds to several types of proteins (target-directed ligand) and zo a compound that binds to a substrate recognition protein. To date, the selective induction of proteolysis (chemical knockdown) by low-molecular-weight compounds via ubiquitin ligase has been studied.
In recent years, a technique in which an artificial complex of E3 [VHL, Cereblon (CRBN), Cellular Inhibitor of Apoptosis Proteinl (cIAPl)] having ubiquitin ligase activity and a target protein is formed in the cell by using a compound in which a ligand for E3 and an inhibitor of the target protein are linked, and degradation of the target protein is induced using the ubiquitin-proteasome system, which is an intracellular protein degradation mechanism has been attracting attention as a new drug discovery technique (Non Patent Literature 1).
Patent Literature 1 discloses a bifunctional compound having, at one end, a VHL ligand that binds to VHL, which is a substrate recognition protein for ubiquitin ligase, and, at the other end, a portion that binds to a target protein.
Patent Literature 2 discloses a bifunctional compound having, at one end, a CRBN ligand that binds to CRBN, which is a substrate recognition protein for ubiquitin ligase, and, at the other end, a portion that binds to a target protein.
Patent Literature 3 discloses a bifunctional compound having a proline compound as a ligand for the substrate recognition protein IAP of the ubiquitin ligase complex.
As described above, while selective induction of the degradation of target protein by bifunctional compounds has been studied, the range of applicable target proteins and the effect of degradation induction have not been necessarily sufficient. Therefore, research and development of bifunctional compounds by using low-molecular-weight compounds with higher effectiveness has been desired.
The present inventors have conducted intensive studies in an attempt to solve the aforementioned problems, and completed the present invention. The present invention is as described below.
[1] A compound represented by the following structural formula (I):
wherein E, X, W, L, and A are as defined above, and X1 and W1 are each a divalent group induced from X and W, or a pharmacologically acceptable salt thereof.
[3] The compound of the above-mentioned [1] or [2], wherein W is an optionally substituted fused heterocyclic group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 6 to 10 ring-constituting atoms,
or a pharmacologically acceptable salt thereof.
[4] The compound of the above-mentioned [1] to [3], wherein W is a group represented by the formula:
wherein
—CO—R9a—W9a—Y9b—W9b
—CO—R9a—W9a—Y9—W9
Y9b is a group represented by:
—CO—R9a—W9a—Y9b—W9b
The present invention relates to a target protein degradation-inducing compound that is a bifunctional compound having a portion that binds to VHL, which is a substrate recognition protein of a ubiquitin ligase complex, at one end and a portion that binds to a target protein at the other end. The target protein degradation-inducing compound of the present invention can exert a wide range of pharmacological activities by regulating the ubiquitination of the target protein and inducing the degradation of the target protein.
The present invention is explained in detail in the following.
The definition of each group used in the present specification is described in detail in the following. Unless particularly noted, each group has the following definition.
In the present specification, when the number of carbon atoms constituting a certain group is indicated, it is indicated at times as “C1-C6” instead of “1 to 6 carbon atoms”. When the number of atoms constituting a certain ring is indicated, it may be indicated as “3- to 10-membered” instead of “3 to 10 atoms constituting the ring”.
When compound (I) has an acidic functional group and/or a basic functional group within the compound, it can form a salt. Examples of such salt include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like.
In the present specification, examples of the “halogen atom” include fluorine, chlorine, bromine, and iodine.
In the present specification, as the “alkyl group” (including the “alkyl” moiety in the definition), an alkyl group having 1 to 10 carbon atoms can be mentioned. Examples thereof include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl, heptyl, octyl, 3-methyloctyl, nonyl, and decyl.
A more preferred embodiment is an alkyl group having 1 to 8 carbon atoms. Another preferred embodiment is an alkyl group having 1 to 6 carbon atoms. A still another embodiment is an alkyl group having 1 to 4 carbon atoms.
In the present specification, as the “alkenyl group” (including the “alkenyl” moiety in the definition), an alkenyl group having 2-6 carbon atoms can be mentioned. Examples thereof include ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 4-methyl-3-pentenyl, 1-hexenyl, 3-hexenyl, and 5-hexenyl.
In the present specification, as the “alkynyl group” (including the “alkynyl” moiety in the definition), an alkynyl group having 2-6 carbon atoms can be mentioned. Examples thereof include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl-1Hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, and 4-methyl-2-pentynyl.
In the present specification, as the “alkylene group” (including the “alkylene” moiety in the definition), an alkylene group having 1 to 6 carbon atoms can be mentioned. Examples thereof include —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, —(CH2)5—, —(CH2)6—, —CH(CH3)—, —C(CH3)2—, —CH(C2H5)—, —CH(C3H7)—, —CH(CH(CH3)2)—, —(CH(CH3))2—, —CH2—CH(CH3)—, —CH(CH3) —CH2—, —CH2—CH2—C(CH3)2—, —C(CH3)2—CH2—CH2—, —CH2—CH2—CH2—C(CH3)2—, and —C(CH3)2—CH2—CH2—CH2—.
In the present specification, as the “alkenylene group” (including the “alkenylene” moiety in the definition), an alkenylene group having 2-6 carbon atoms can be mentioned. Examples thereof include —CH═CH—, —CH2—CH═CH—, —CH═CH—CH2—, —C(CH3)2—CH═CH—, —CH═CH—C(CH3)2—, —CH2—CH═CH—CH2—, —CH2—CH2—CH═CH—, —CH═CH—CH2—CH2—, —CH═CH—CH═CH—, —CH═CH—CH2—CH2—CH2—, and —CH2—CH2—CH2—CH═CH—.
In the present specification, as the “alkynylene group” (including the “alkynylene” moiety in the definition), an alkynylene group having 2-6 carbon atoms can be mentioned. Examples thereof include —C≡C—, —CH2—C≡C—, —C≡C—CH2—, —C(CH3)2—C≡C—, —C≡C—C(CH3)2—, —CH2—C≡C—CH2—, —CH2—CH2—C≡C—, —C≡C—CH2—CH2—, —C≡C—C≡C—, —C≡C—CH2—CH2—CH2—, and —CH2—CH2—CH2—C≡C—.
In the present specification, as the “alkoxy group” (including the “alkoxy” moiety in the definition), an alkoxy group having 1 to 6 carbon atoms can be mentioned. Examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, and hexyloxy.
A more preferred embodiment is an alkoxy group having 1 to 4 carbon atoms.
In the present specification, as the “alkoxycarbonyl group”, an (alkoxy having 1 to 6 carbon atoms)-carbonyl group can be mentioned. Examples thereof include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, and hexyloxycarbonyl.
In the present specification, as the “cycloalkyl group” (including the “cycloalkyl” moiety in the definition), a monocycle or condensed cycloalkyl group having 3 to 10 (preferably, 3 to 8) carbon atoms can be mentioned. Examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, and adamantyl.
A more preferred embodiment is a cycloalkyl group having 3 to 6 carbon atoms.
In the present specification, as the “cycloalkyl divalent group” (at times referred to as “cycloalkane-diyl group”) (including the “cycloalkyl divalent group” moiety in the definition), a divalent group formed because the “cycloalkyl group having 3 to 10 carbon atoms” explained above has another bond can be mentioned, for example, 1,3-cyclopropane-diyl.
A more preferred embodiment is a cycloalkyl divalent group having 3 to 6 carbon atoms.
In the present specification, as the “cycloalkyloxy group”, a (cycloalkyl having 3 to 10 carbon atoms)-oxy group can be mentioned. Examples thereof include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and cycloheptyloxy, cyclooctyloxy.
In the present specification, as the “aryl group” (including the “aryl” moiety in the definition), a monocyclic or condensed aryl group having 6 to 14 carbon atoms can be mentioned. Examples thereof include phenyl, 1-naphthyl, 2-naphthyl, 1-anthryl, 2-anthryl, and 9-anthryl.
A more preferred embodiment is an aryl group having 6 to carbon atoms.
In the present specification, examples of the “C7-16 aralkyl group” (including the “C7-16 aralkyl” moiety in the definition) include benzyl, phenethyl, naphthylmethyl, and phenylpropyl.
In the present specification, as the “aryl divalent group” (including the “aryl divalent group” moiety in the definition) (at times referred to as “arylene group”), a divalent group formed because the “aryl group” explained above has another bond can be mentioned. Examples thereof include an aryl divalent group having 6 to 14 carbon atoms (preferably, aryl divalent group having 6 to 10 carbon atoms), for example, phenylene.
In the present specification, as the “heterocyclic group” (including the “heterocycle” moiety in the definition) (at times referred to as “non-aromatic heterocyclic group”), a monocyclic or condensed heterocyclic group (non-aromatic heterocyclic group) containing 1 to 6 (preferably 1 to 4) same or different hetero atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom as ring-constituting atoms besides carbon atom, and 3 to 14 (preferably 3 to 10) ring-constituting atoms, can be mentioned.
Preferred examples of the “heterocyclic group” (“non-aromatic heterocyclic group”) include a monocyclic heterocyclic group having 3 to 8 ring-constituting atoms (non-aromatic heterocyclic group) such as aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, tetrahydrothienyl, tetrahydrofuranyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, oxazolinyl, oxazolidinyl, pyrazolinyl, pyrazolidinyl, thiazolinyl, thiazolidinyl, tetrahydroisothiazolyl, tetrahydrooxazolyl, tetrahydroisoxazolylpiperidinyl, piperazinyl, tetrahydropyridinyl, dihydropyridinyl, dihydrothiopyranyl, tetrahydropyrimidinyl, tetrahydropyridazinyl, dihydropyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, thiomorpholinyl, azepanyl, diazepanyl, azepinyl, oxepanyl, azocanyl, diazocanyl, and the like; and fused polycyclic (preferably bi or tricyclic)heterocyclic group (non-aromatic heterocyclic group) having 9 to 14 ring-constituting atoms such as dihydrobenzofuranyl, dihydrobenzoimidazolyl, dihydrobenzooxazolyl, dihydrobenzothiazolyl, dihydrobenzoisothiazolyl, dihydronaphto[2,3-b]thienyl, tetrahydroisoquinolyl, tetrahydroquinolyl, 4H-quinolizinyl, indolinyl, isoindolinyl, tetrahydrothieno[2,3-c]pyridinyl, tetrahydrobenzoazepinyl, tetrahydroquinoxalinyl, tetrahydrophenanthridinyl, hexahydrophenothiazinyl, hexahydrophenoxazinyl, tetrahydrophthalazinyl, tetrahydronaphthyridinyl, tetrahydroquinazolinyl, tetrahydrocinnolinyl, tetrahydrocarbazolyl, tetrahydro-R-carbolinyl, tetrahydroacrydinyl, tetrahydrophenazinyl, tetrahydrothioxanthenyl, octahydroisoquinolyl, and the like.
In the present specification, the “heterocyclic group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 3 to 10 ring-constituting atoms” refers to the “heterocyclic group” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and having 3 to 10 ring-constituting atoms.
In the present specification, the “fused heterocyclic group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 6 to 10 ring-constituting atoms” refers to the “heterocyclic group” explained above which is a fused polycyclic group containing 1 to 6 hetero atoms as ring-constituting atoms and having 6 to 10 ring-constituting atoms.
In the present specification, the “heterocyclic group having 3 to 6 ring-constituting atoms” refers to the “heterocyclic group” explained above and having 3 to 6 ring-constituting atoms.
In the present specification, the “heterocyclic group containing a nitrogen atom and 1 to 5 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 3 to 10 ring-constituting atoms” refers to the “heterocyclic group” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and at least one nitrogen atom as a ring-constituting atom, and having 3 to 10 ring-constituting atoms.
In the present specification, the “heterocyclic group containing 1 or 2 oxygen atoms, and having 5 or 6 ring-constituting atoms” refers to the “heterocyclic group” explained above and containing 1 or 2 oxygen atoms, and having or 6 ring-constituting atoms.
In the present specification, as the “heterocyclic divalent group” (including the “heterocyclic divalent group” moiety in the definition), a divalent group formed because the “heterocyclic group” explained above has another bond can be mentioned, for example, piperazine-diyl.
In the present specification, the “heterocyclic divalent group having 3 to 6 ring-constituting atoms” refers to the “heterocyclic divalent group” explained above and having 3 to 6 ring-constituting atoms.
In the present specification, the “heterocyclic divalent group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 3 to 10 ring-constituting atoms” refers to the “heterocyclic divalent group” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and having 3 to 10 ring-constituting atoms.
In the present specification, the “heterocyclic divalent group containing 1 to 4 nitrogen atoms, and having 5 or 6 ring-constituting atoms” refers to the “heterocyclic divalent group” explained above and containing 1 to 4 nitrogen atoms, and having 5 or 6 ring-constituting atoms.
In the present specification, the “monocyclic heterocyclic divalent group containing 1 or 2 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 4 to 7 ring-constituting atoms” refers to the “heterocyclic divalent group” explained above and containing 1 or 2 hetero atoms and having 4 to 7 ring-constituting atoms.
In the present specification, as the “heteroaryl group” (including the “heteroaryl” moiety in the definition) (at times referred to as “aromatic heterocyclic group”), a monocyclic or condensed heteroaryl group (aromatic heterocyclic group) containing 1 to 6 (preferably 1 to 4) same or different hetero atom selected from a nitrogen atom, an oxygen atom, and a sulfur atom as ring-constituting atoms besides carbon atom, and having 5 to 14 (preferably 5 to 10) ring-constituting atoms can be mentioned.
Preferred examples of the “heteroaryl group” (“aromatic heterocyclic group”) include a monocyclic heteroaryl group (aromatic heterocyclic group) having 5 or 6 ring-constituting atoms, such as thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and the like; and a fused polycyclic (preferably bi or tricyclic)heteroaryl group (aromatic heterocyclic group) having 8 to 14 ring-constituting atoms, such as benzothiophenyl, benzofuranyl, benzoimidazolyl, benzooxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl, imidazopyridinyl, thienopyridinyl, furopyridinyl, pyrrolopyridinyl, pyrazolopyridinyl, oxazolopyridinyl, thiazolopyridinyl, imidazopyrazinyl, imidazopyrimidinyl, thienopyrimidinyl, furopyrimidinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, oxazolopyrimidinyl, thiazolopyrimidinyl, pyrazolotriazinyl, naphtho[2,3-b]thienyl, phenoxathiinyl, indolyl, isoindolyl, 1H-indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, carbazolyl, p-carbolinyl, phenanthridinyl, acrydinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.
In the present specification, the “heteroaryl group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 5 to 10 ring-constituting atoms” refers to the “heteroaryl group” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and having 5 to 10 ring-constituting atoms.
In the present specification, the “heteroaryl group having 5 ring-constituting atoms” refers to the “heteroaryl group” explained above and having 5 ring-constituting atoms.
In the present specification, the “heteroaryl group containing 1 to 4 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 5 or 6 ring-constituting atoms” refers to the “heteroaryl group” explained above and containing 1 to 4 hetero atoms as ring-constituting atoms and having 5 or 6 ring-constituting atoms.
In the present specification, as the “heteroaryl divalent group” (including the “heteroaryl divalent group” moiety in the definition), a divalent group formed because the “heteroaryl group” explained above has another bond can be mentioned, for example, pyridine-diyl.
In the present specification, the “heteroaryl divalent group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 5 to 10 ring-constituting atoms” refers to the “heteroaryl divalent group” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and having 5 to 10 ring-constituting atoms.
In the present specification, the “heteroaryl divalent group containing 1 to 4 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 5 or 6 ring-constituting atoms” refers to the “heteroaryl divalent group” explained above and containing 1 to 4 hetero atoms and having 5 or 6 ring-constituting atoms.
In the present specification, as the “aromatic hydrocarbocycle”, an aromatic hydrocarbocycle having 6 to 14 (preferably, 6 to 10) carbon atoms can be mentioned. Examples thereof include benzene and naphthalene.
In the present specification, as the “non-aromatic hydrocarbocycle”, a saturated or partially unsaturated cyclic hydrocarbocycle can be mentioned. Examples thereof include cycloalkane and cycloalkene.
In the present specification, as the “cycloalkane”, “C3-10 cycloalkane” can be mentioned. Examples thereof include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, and cyclodecane.
In the present specification, as the “cycloalkene”, “C3-10 cycloalkene” can be mentioned. Examples thereof include cyclopropene, cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclooctene, cyclononene, and cyclodecene.
In the present specification, the “non-aromatic hydrocarbocycle having 5 or 6 ring-constituting atoms” refers to the “non-aromatic hydrocarbocycle” explained above and having 5 or 6 ring-constituting atoms.
In the present specification, as the “aromatic heterocycle”, an aromatic heterocycle containing 1 to 6 (preferably 1 to 4) hetero atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom as ring-constituting atoms besides carbon atom, and having 5 to 14 (preferably 5 to 10) ring-constituting atoms can be mentioned. Preferred examples of the “aromatic heterocycle” include monocyclic aromatic heterocycle having 5 or 6 (5- or 6-membered) ring-constituting atoms such as thiophene, furan, pyrrole, imidazole, pyrazole, thiazole, isothiazole, oxazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole, triazole, tetrazole, triazine and the like; (8- to 14-membered) fused polycyclic (preferably bi or tricyclic) aromatic heterocycle having 8 to 14 ring-constituting atoms such as benzothiophene, benzofuran, benzoimidazole, benzoxazole, benzoisoxazole, benzothiazole, benzoisothiazole, benzotriazole, imidazopyridine, thienopyridine, furopyridine, pyrrolopyridine, pyrazolopyridine, oxazolopyridine, thiazolopyridine, imidazopyrazine, imidazopyrimidine, thienopyrimidine, furopyrimidine, pyrrolopyrimidine, pyrazolopyrimidine, oxazolopyrimidine, thiazolopyrimidine, pyrazolopyrimidine, pyrazolotriazine, naphtho[2,3-b]thiophene, phenoxathiine, indole, isoindole, 1H-indazole, purine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, carbazole, β-carboline, phenanthridine, acridine, phenazine, phenothiazine, phenoxathiine and the like.
In the present specification, the “aromatic heterocycle containing a nitrogen atom, and further optionally containing 1 to 5 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom” refers to the “aromatic heterocycle” explained above and containing 1 to 6 hetero atoms as ring-constituting atoms and having at least one nitrogen atom as the ring-constituting atom.
In the present specification, as the “heterocycle”, for example, a non-aromatic heterocycle containing 1 to 6 (preferably, 1 to 4) hetero atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom as ring-constituting atoms besides carbon atom, and having 3 to 14 (preferably 3 to 10) ring-constituting atoms can be mentioned. Preferred examples of the “heterocycle” (“non-aromatic heterocycle”) include a monocyclic non-aromatic heterocycle having 3 to 8 (3- to 8-membered) ring-constituting atoms such as aziridine, oxirane, thiirane, azetidine, oxetane, thietane, tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, imidazoline, imidazolidine, oxazoline, oxazolidine, pyrazoline, pyrazolidine, thiazoline, thiazolidine, tetrahydroisothiazole, tetrahydrooxazole, tetrahydroisoxazole, piperidine, piperazine, tetrahydropyridine, dihydropyridine, dihydrothiopyran, tetrahydropyrimidine, tetrahydropyridazine, dihydropyran, tetrahydropyran, tetrahydrothiopyran, morpholine, thiomorpholine, azepane, diazepane, azepine, azocane, diazocane, oxepane and the like; and a fused polycyclic (preferably bi or tricyclic) non-aromatic heterocycle having 9 to 14 (9- to 14-membered) ring-constituting atoms such as dihydrobenzofuran, dihydrobenzoimidazole, dihydrobenzooxazole, dihydrobenzothiazole, dihydrobenzoisothiazole, dihydronaphto[2,3-b]thiophene, tetrahydroisoquinoline, tetrahydroquinoline, 4H-quinolizine, indoline, isoindoline, tetrahydrothieno[2,3-c]pyridine, tetrahydrobenzoazepine, tetrahydroquinoxaline, tetrahydrophenanthridine, hexahydrophenothiazine, hexahydrophenoxathiine, tetrahydrophthalazine, tetrahydronaphthyridine, tetrahydroquinazoline, tetrahydrocinnoline, tetrahydrocarbazole, tetrahydro-β-carboline, tetrahydroacridine, tetrahydrophenazine, tetrahydrothioxanthene, octahydroisoquinoline and the like.
In the present specification, the “heterocycle containing a nitrogen atom, and further optionally containing 1 to 5 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom” refers to the “heterocycle” explained above and containing 1 to 6 hetero atoms as the ring-constituting atoms and having at least one nitrogen atom as the ring-constituting atom.
In the present specification, the “heterocycle containing a nitrogen atom, and further optionally containing 1 to 3 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 5 or 6 ring-constituting atoms” refers to the “heterocycle” explained above and containing 1 to 4 hetero atoms as the ring-constituting atoms and containing at least one nitrogen atom as the ring-constituting atom and having 5 or 6 ring-constituting atoms.
In the present specification, the “heterocycle containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 3 to 10 ring-constituting atoms” refers to the “heterocycle” explained above and containing 1 to 6 hetero atoms as the ring-constituting atoms and having 3 to 10 ring-constituting atoms.
In the present specification, as the “substituent” of the “optionally substituted group and ring” in the “optionally substituted alkyl group”, “optionally substituted cycloalkyl group”, “optionally substituted aryl group”, “optionally substituted heterocyclic group”, “optionally substituted heteroaryl group”, “optionally substituted fused heterocyclic group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 6 to 10 ring-constituting atoms”, “optionally substituted heteroaryl group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 5 to 10 ring-constituting atoms”, and other definitions relating to compound (I), the substituents selected from the following [Substituent Group] can be mentioned.
1 to 5 (preferably 1 to 3) “substituents” may be present at substitutable positions. When the number of the substituents is two or more, each substituent may be the same or different.
When specific explanations on the “substituent” of each group, ring, and the like are given, such explanations are to be followed.
In the present specification, the “chemical linker” can be determined appropriately by those of ordinary skill in the art according to the selection of the target protein. The detail is described later.
In the present specification, the “target-directed ligand” is, for example, a group having a portion capable of binding to the “target protein” or a portion that binds to the “target protein”. The detail is described later.
The compound of the present invention is a compound (I) represented by the following structural formula (I) or a pharmacologically acceptable salt thereof.
In compound (I), L is bonded to either W or X. Therefore, compound (I) may structurally take the following two embodiments. These embodiments are all encompassed in the scope of the present invention.
Each symbol of compound (I) is described in detail below.
E is a bond, —CO—, —SO—, or —SO2—, preferably, a bond or —CO—, more preferably —CO—.
In another preferred embodiment, X is a group represented by —CHRm—X1—X2, wherein Rm is
The groups (W1) to (W4) for W are described in detail below.
The “aryl group” of the “optionally substituted aryl group” is preferably an aryl group having 6 to 14 carbon atoms, more preferably, an aryl group having 6 to 10 carbon atoms.
The aryl group is optionally substituted at substitutable position(s) by 1 to 3 same or different substituents selected from the “Substituent Group” explained in the aforementioned “Definition of each group used in the present specification”.
Group (W2): An Optionally Substituted Fused Heterocyclic Group Containing 1 to 6 Same or Different Atom Selected from Nitrogen Atom, Oxygen Atom, and Sulfur Atom, and Having 6 to 10 Ring-Constituting Atoms
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of the “fused heterocyclic group” of the “optionally substituted fused heterocyclic group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 6 to 10 ring-constituting atoms. Preferably, both an embodiment in which two 6-membered rings are condensed, and an embodiment in which a 5-membered ring and a 6-membered ring are condensed are encompassed.
The “fused heterocyclic group” is optionally substituted at substitutable position(s) by 1 to 3 same or different substituents selected from the “Substituent Group” explained in the aforementioned “Definition of each group used in the present specification”.
The former encompasses, for example, tetrahydroquinoline, tetrahydroquinazoline, and the like.
The latter is more specifically explained below. The latter is preferably a group represented by the formula (W2a):
The “optionally substituted heteroaryl group having five ring-constituting atoms, and two A1 are each independently a group or atom selected from ═CRZ1—, ═N—, —NRZ2—, —O—, and —S—, two A2 are each independently a carbon atom or a nitrogen atom” for group A is, for example, pyrrolyl, imidazolyl, thienyl, furyl, oxazolyl, thiazolyl, triazolyl, tetrazolyl, or the like.
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of each group of the “aromatic hydrocarbocycle”, “aromatic heterocycle optionally containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom”, “non-aromatic hydrocarbocycle”, and “heterocycle optionally containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom” in ring Q.
The aforementioned “Definition of each group used in the present specification” can be also referred to for the meaning of each group of the “halogen atom”, “optionally substituted alkyl group”, “optionally substituted cycloalkyl group”, “optionally substituted alkoxy group”, “optionally substituted cycloalkyloxy group”, “—CO—N(R7a)(R7b)”, “—N(R7a)(R7b)”, “—N(R7c)—CO—R7d”, and “—CO—R7e,”, which are the substituents used for substituting the above-mentioned “aromatic hydrocarbocycle” and the like in ring Q.
Group (W2) is more preferably a group represented by the formula (W2b):
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of the “heteroaryl group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 3 to 10 ring-constituting atoms” and the “substituent” used to optionally substitute the heteroaryl group.
The group (W3) is preferably a group represented by the following formula (W3a):
The “group A′” in the group represented by the formula (W3a) is preferably a group selected from imidazolyl group, pyrazolyl group, thiazolyl group (e.g., 1,2-thiazolyl, 1,3-thiazolyl), isothiazolyl group, thiadiazolyl group, isothiadiazolyl group, oxazolyl group, isoxazolyl group, oxadiazolyl group, triazolyl group (1,2,3-triazolyl, 1,2,4-triazolyl), and tetrazolyl group, further preferably, a group selected from imidazolyl group, thiazolyl group, and oxazolyl group.
The aforementioned “Definition of each group used in the present specification” can be referred to for the meanings of Ra1, Ra2 and Ra3 in the group represented by the formula (W3a).
The group (W3) is more preferably a group represented by the following formula (W3b) or (W3c):
R6a1, R6a2, and R6a3 in the above-mentioned formula are explained in detail below.
More preferably, R6a1 is
Further preferably, R6a1 is
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of each group.
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of each group.
Further preferably, R6a3 is
—CO—R9a—W9a—Y9b—W9b
Further more preferably, R6a3 is
—CO—R9a—W9a—Y9b—W9b
wherein
The group (W4) is preferably an alkyl group having 1 to 6 carbon atoms and optionally substituted by 1 to 3 groups selected from 1) a halogen atom, 2) a hydroxy group, 3) an alkoxy group having 1 to 6 carbon atoms, and 4) an optionally substituted heterocyclic group containing 1 to 6 atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 3 to 10 ring-constituting atoms.
L is a bond or a chemical linker, preferably, a group represented by the formula (L-I): -(La)q- wherein q is an integer of 1 to 100 (more preferably, an integer of 1 to 50), and
La in the number of q are each independently a group selected from
(CH2)p1a—O—(CH2—CH2—O)p2a—(CH2)p3a
The aforementioned “Definition of each group used in the present specification” can be referred to for the meaning of the “optionally substituted cycloalkyl divalent group”, “optionally substituted heterocyclic divalent group”, “optionally substituted aryl divalent group”, “optionally substituted heteroaryl divalent group”, “halogen atom”, “optionally substituted alkyl group”, “optionally substituted cycloalkyl group”, “optionally substituted aryl group”, “optionally substituted heteroaryl group”, “optionally substituted C1-C8 alkyl group”, and “optionally substituted C3-C8 cycloalkyl group” in the above-mentioned definitions.
A is a target-directed ligand, more specifically, a group having a moiety capable of binding to a target protein or a moiety that binds to the target protein. For example, a preferred specific example of the target-directed ligand is a compound that binds to the following target protein. The “target-directed ligand” of the present invention is not limited thereto.
Those of ordinary skill in the art can appropriately select or design and use a target-directed ligand according to the target protein of interest. Those described in Examples 163 to 165 described later are the examples thereof.
In the present invention, the target protein bound to A is preferably a protein having a biological function selected from the group consisting of structure, regulation, hormone, enzyme, gene regulation, immunity, contraction, storage, transport, and signal transduction. The protein is more preferably selected from the group consisting of structural protein, receptor, enzyme, cell surface protein, and proteins related to integrated cellular function, including proteins involved in catalytic activity, aromatase activity, motor activity, helicase activity, metabolic process (anabolism and catabolism), antioxidant activity, proteolysis, biosynthesis, kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulatory factor activity, signal transducer activity, structural molecular activity, binding activity (protein, lipid carbohydrates), receptor activity, cellular motility, membrane fusion, intercellular signal transduction, control of biological processes, development, cell differentiation, stimulus response, cell adhesion, cell death, transport(protein transporter activity, nuclear transport, transporter activity, channel transporter activity, carrier activity, permease activity, secretion activity, electron transporter activity), pathogenesis, virus outer shell, chaperon regulatory factor activity, nucleic acid binding activity, transcriptional regulator activity, epigenetics control, aggregation, extracellular organization, biogenetic activity, or translation regulatory factor activity.
In the present invention, the target protein bound to A is preferably selected from the group consisting of proteins related to cancer related proteins, autoimmune disease related proteins, inflammatory disease related proteins, neurodegenerative disease related proteins, muscular disease related proteins, sensory system disease related proteins, circulatory disease related proteins, metabolic disease related proteins, and genetic disease related proteins.
When compound (I) contains optical isomers (enantiomer, diastereomer), stereoisomer, regioisomer, and rotamer, these are also included as compound (I), and can be respectively obtained as single products by synthesis methods and separation methods (e.g., concentration, solvent extraction, column chromatography, recrystallization, etc.) known per se.
Compound (I) may be converted to a prodrug as appropriate, and such embodiment is also included in the scope of the present invention (hereinafter compound (I) and a prodrug thereof are at times collectively referred to as “the compound of the present invention”). The prodrug may be one that changes to compound (I) under physiological conditions, such as the one described in “Drug Development” Vol. 7 “Molecular Design”, p. 163-198 published by HIROKAWA SHOTEN (1990).
Compound (I) may be any of hydrate, non-hydrate, solvate, and non-solvate. In addition, compound (I) may be a compound labeled or substituted with an isotope (e.g., 2H, 3H, 11C, 14C, 18F, 35S, 125I, etc.). A compound labeled or substituted with an isotope can be used as a tracer (PET tracer) used in positron emission tomography (PET), and may be useful in the fields of medical diagnosis and the like. A deuterium conversion form wherein H is converted to 2H(D) is also encompassed in compound (I).
Tautomers are also encompassed in compound (I).
The production method of compound (I) is explained below.
The compounds of the present invention can be produced by the following methods A to X. While these methods and steps may be combined but the production method thereof is not limited thereto.
The formula (I) of the present invention:
wherein each symbol is as defined above.
The formula (1) of the intermediate of the present invention:
wherein PG1 is an amine-protecting group, PG2 is a hydroxy-protecting group, and other symbols are as defined above.
As the protecting group represented by PG1, a carbamate-protecting group can be mentioned. As the protecting group represented by PG2, a silyl-protecting group can be mentioned.
When PG1 is, for example, a Boc group, deprotection can be performed in a suitable solvent in the presence of an acid. When PG2 is, for example, a TBS group, deprotection can be performed in a suitable solvent in the presence of an acid, base, or fluoride ion.
The production method described here is suitable for producing an intermediate of a compound represented by the formula (I) wherein W is an imidazolyl group, namely, the following compound [A-2].
wherein each symbol is as defined above.
The formula [A-2] can be derived from the formula [A-1] according to a known method (e.g., BIOORGANIC & Medicinal Chemistry Letters, 26(21), 5354-5360; 2016).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an imidazolyl group, and Ra2 and Ra3 are substituted, i.e., the following compound [B-2].
wherein each symbol is as defined above.
The formula [B-2] can be derived from the formula [B-1] according to a known method (e.g., WO2006099060).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an imidazolyl group, Ra1 and Ra2 are each a hydrogen atom, and Ra3 is substituted by carboxylic acid, i.e., the following compound [C-4].
wherein RCO[MC] is an alkali metal alkoxide, and each symbol is as defined above.
The formula [C-1] can be derived from compound [A-1] according to a known method (e.g., Journal of Medicinal Chemistry, 33(1), 317-27; 1990).
The formula [C-2] is obtained by decomposing the trifluoro group in the formula [C-1] with a base. The reaction proceeds using alkali metal alkoxide in an appropriate alcohol solvent generally from room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally 5 min to 12 hr. Examples of the base having an alkoxy group include alkali metal alkoxide such as sodium methoxide, specifically potassium methoxide, sodium methoxide, lithium methoxide, sodium ethoxide, potassium tert-butoxide, and the like. Examples of the solvent include methanol, ethanol, tert-butanol and the like.
The formula [C-3] is obtained by decomposition of the formula [C-2] under acidic conditions. The reaction proceeds using an acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, the reaction generally proceeds in 1 hr to 24 hr. Examples of the acid include trifluoroacetic acid, hydrochloric acid, and the like. Examples of the solvent include dichloromethane, 1,2-dichloroethane, chloroform, methanol, ethyl acetate, toluene, 1,4-dioxane, and the like.
The formula [C-4] is obtained by subjecting the formula [C-3] to a basic condition or an acidic condition. The reaction proceeds using a base or acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, the reaction generally proceeds in 1 hr to 48 hr. When a base is used, sodium hydroxide, potassium hydroxide, lithium hydroxide, and the like can be mentioned, and when an acid is used, trifluoroacetic acid, hydrochloric acid, and the like can be mentioned. When a base is used as a solvent, for example, methanol, ethanol, tetrahydrofuran, and water can be mentioned, and when an acid is used, dichloromethane, 1,2-dichloroethane, chloroform, methanol, ethyl acetate, toluene, 1,4-dioxane, water and the like can be mentioned.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a thiazolyl group substituted by Ra2 and Ra3′ i.e., the following compound [D-4].
wherein XA is a halogen atom, each symbol is as defined above, and the halogen atom is a chlorine atom, a bromine atom, or an iodine atom.
The formula [D-2] is obtained by thioamidating the formula [D-1]. The thioamidation reaction proceeds using a sulfurizing agent in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally 0.5 hr to 24 hr. Examples of the sulfurizing agent include Lawesson's reagent, diphosphorus pentasulfide, and the like. Examples of the solvent include 1,2-dichloroethane, tetrahydrofuran, 1,4-dioxane and the like.
The formula [D-4] can be derived from the formula [D-2] and the formula [D-3] according to a known method (e.g., ChemBioChem, 12(15), 2284-2288; 2011, WO 2009098448, WO 2010060952).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an oxadiazolyl group substituted by Ra3′ i.e., the following compound [E-3].
wherein each symbol is as defined above.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a triazolyl group substituted by Ra1 and Ra3′ i.e., the following compound [F-3].
wherein each symbol is as defined above.
The formula [F-1] can be derived from the formula [D-1] according to a known method (e.g., Tetrahedron Letters, 45(52), 9557-9559; 2004).
The formula [F-3] can be derived from the formula [F-1] and the formula [F-2] according to a known method (e.g., US20120264735).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a tetrazolyl group, i.e., the following compound [G-2].
wherein each symbol is as defined above.
The formula [G-2] can be derived from the formula [G-1] according to a known method (e.g., Synthesis, 46(15), 2065-2070; 2014, WO 2011035900).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) W is triazolyl group, substituted by Ra2, i.e., the following compound [H-3].
wherein each symbol is as defined above.
The formula [H-1] can be derived from the formula [A-1] according to a known method (e.g., Bioorganic & Medicinal Chemistry Letters, 26(5), 1419-1427; 2016).
The formula [H-3] can be derived from the formula [H-1] and the formula [H-2] according to a known method (e.g., Organometallics, 30(5), 1021-1029; 2011).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a pyrazolyl group substituted by Ra2 and Ra3, i.e., the following compound (I-3).
wherein each symbol is as defined above.
The formula [I-3] can be derived from the formula [I-1] and the formula [I-2] according to a known method (e.g., WO 2002034716).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an oxazolyl group substituted by Ra2 and Ra3, i.e., the following compound [J-2].
wherein each symbol is as defined above.
The formula [J-2] can be derived from the formula [J-1] according to a known method (e.g., WO 2009080226).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a pyridyl group, i.e., the following compound [K-2].
wherein each symbol is as defined above.
The formula [K-2] can be derived from the formula [K-1] according to a known method (e.g., WO 2013061977).
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an alkyl group optionally substituted by a group selected from a heterocyclic group and substituted by X2aL, i.e., the following compound [L-5].
wherein X2aL is a carbon or oxygen atom, and each symbol is as defined above.
The formula [L-2] is obtained by the following two steps.
The formula [L-2] is obtained by a reduction reaction of the formula [L-1]. The reaction proceeds using a reducing agent in an appropriate solvent generally from −78° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, the reaction proceeds generally for 10 min to 24 hr. When a reducing agent is used, hydrogenated lithium aluminum hydride, DIBAL-H, and the like can be mentioned. Examples of the solvent include methylene chloride, diethyl ether, tetrahydrofuran, and toluene.
The formula [L-2] is obtained by a reduction reaction of the formula [E-1]. The reaction proceeds using a reducing agent in an appropriate solvent generally from −78° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, the reaction generally proceeds for 10 min to 24 hr. When a reducing agent is used, hydrogenated lithium aluminum hydride, BH3·THF, and the like can be mentioned. Examples of the solvent include diethyl ether and tetrahydrofuran.
The formula [L-3] is obtained by halogenation of the formula [L-2]. The reaction proceeds using a halogenating agent in an appropriate solvent generally from 0° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, the reaction generally proceeds in 10 min to 24 hr. Examples of the halogenating agent include iodine and triphenyl phosphine, or carbon tetrabromide and triphenyl phosphine, and the like. In the case of iodine and triphenyl phosphine, imidazole or the like is used as a base. Examples of the solvent include diethyl ether, tetrahydrofuran, and dichloromethane.
The formula [L-5] is obtained by an alkylation reaction of the formula [L-3] and the formula [L-4]. The alkylation reaction proceeds using a base and a alkylating agent such as halogenated alkyl and the like, in an appropriate solvent generally from 0° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 30 min to 24 hr. Examples of the base include inorganic bases such as sodium hydride, potassium hydroxide, cesium carbonate, potassium carbonate, and the like, alkoxides such as potassium tert-butoxide, sodium methoxide, and the like, and the like. Examples of the solvent include N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and the like.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is a heteroaryl group having 5 ring-constituting atoms, A3c is a carbon atom, substituted by Ra1 and Ra2, further Ra3 is an optionally substituted aryl group, or substituted by an optionally substituted heteroaryl group, i.e., the following compound [M-4].
wherein B1 is boronic acid or boronic acid ester optionally having substituent(s), and other each symbol is as defined above. As used herein, boronic acid ester optionally having substituent(s) for B1 is pinacolatoboron, neopentyl glycolatoboron or the like.
The formula [M-2] is obtained by halogenation of the formula [M-1]. The reaction proceeds in the presence of a halogenating agent in a suitable solvent from 0° C. to solvent refluxing. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally 1 hr to 24 hr. The halogenating agent is, for example, bromine, iodine, NCS, NBS, NIS, or the like. As the solvent, acetonitrile, methylene chloride, carbon tetrachloride, 1,2-dichloroethane, tetrahydrofuran, and the like can be mentioned.
The formula [M-4] is obtained by a coupling reaction of the formula [M-2] and boronic acid derivative [M-3]. The reaction preferably proceeds in the presence of a palladium catalyst, a phosphine ligand, and a base in a suitable solvent from 0° C. to under heating, particularly from room temperature to the boiling point of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally 1 hr to 24 hr. Examples of the palladium catalyst include palladium (II) acetate, palladium (II) chloride, tris(dibenzylideneacetone)dipalladium(0) or chloroform adduct thereof, and the like. Examples the phosphine ligand include triphenylphosphine, 2-(dicyclohexylphosphino)-2′,4′,6′-triisopropyl-1,1′-biphenyl, 2-(dicyclohexylphosphino)-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl, 2-(dicyclohexylphosphino)-2,6-diisopropoxy-1,1′-biphenyl, 2-di-tert-butylphosphino-2′-4′-6′-triisopropylbiphenyl, 2-dicyclohexylphosphino-2′-6′-dimethoxybiphenyl, 2-(dicyclohexylphosphino)-2-(N,N-dimethylamino)biphenyl, tri-ortho-tolylphosphine, 2-(dicyclohexylphosphino)biphenyl, 2-(di-tert-butylphosphino)biphenyl, 2-(di-tert-butylphosphino)-1,1-binaphthyl, tri-tert-butylphosphine, tri-tert-butylphosphonium tetrafluoroborate, and the like. A reagent in which a palladium catalyst and a phosphine ligand form a complex may also be used. Examples thereof include tetrakis(triphenyl phosphine)palladium(0), 1,1-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, dichlorobis(triphenylphosphine)palladium(II), dichlorobis(tricyclohexylphosphine)palladium(II), bis(tri-tert-butylphosphine)palladium(0), chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl) (2′-amino-1,1′-biphenyl-2-yl)palladium(II), [(2-dicyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)palladium(II) methanesulfonate, (2-dicyclohexyl phosphino-2,6-diisopropoxy-1,1′-biphenyl) [2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate, [(2-di-tert-butylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)palladium(II) methanesulfonate, and the like. Examples of the base include tert-butoxy sodium, potassium acetate, tripotassium phosphate, cesium carbonate, potassium carbonate, sodium hydrogen carbonate, triethylamine, diisopropyl ethylamine, dicyclohexylethylamine, potassium fluoride, cesium fluoride, and the like. Examples of the solvent include ether solvents such as tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane and the like, alcoholic solvents such as methanol, ethanol, propanol, butanol, and the like, N,N-dimethylformamide, NMP or a mixed solvent of an organic solvent and water, and the like.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an imidazolyl group, a thiazolyl group, or an oxazolyl group, substituted by R6a2′ R6a3 is —CO—NRa3N6Ra3N6′ or —CO-(a heterocyclic group containing a nitrogen atom, optionally further containing 1 to 5 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom, and having 3 to 10 ring-constituting atoms, which is optionally further substituted in addition to Y9a), i.e., the following compound [N-4] or [N-5].
wherein each symbol is as defined above.
As the method for obtaining the formula [N-4] or [N-5], for example, any of the following three methods is used.
The formula [N-4] or [N-5] is obtained by a condensation reaction of the formula [N-1] and the formula [N-2] or the formula [N-3]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 30 min to 24 hr. Examples of the condensing agent include 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (WSC·HCl), 1-[bis (dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 4-(4,6-dimethoxy[1.3.5]triazin-2-yl)-4-methylmorpholinium chloride hydrate (DMT-MM), 2-chloro-1-methylpyridinium iodide, and the like. Examples of the solvent include methanol, N,N-dimethylformamide, chloroform, dichloromethane, tetrahydrofuran, and the like. The reaction may be promoted by adding 1-hydroxybenzotriazole (HOBt). Examples of the base include triethylamine, N,N-diisopropyl ethylamine, pyridine, and the like.
The formula [N-4] or [N-5] is obtained by converting the formula [N-1] to an acid halide with a halogenating agent, and then reacting same with the formula [N-2] or the formula [N-3]. The reaction proceeds using a base in an appropriate solvent at generally from −20° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 0.5 hr to 24 hr. Examples of the halogenating agent include thionyl chloride, oxalyl chloride, phenylphosphonyl dichloride, and the like. Examples of the base include triethylamine, pyridine and the like. Examples of the solvent include dichloromethane, 1,2-dichloroethane, chloroform, pyridine, toluene, and the like.
The formula [N-4] or [N-5] is obtained by converting the formula [N-1] to a mixed acid anhydride, and then reacting same with the formula [N-2] or the formula [N-3]. The reaction proceeds using a base in an appropriate solvent at generally from −20° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 0.5 hr to 24 hr. Examples of the reagent to form the mixed acid anhydride include acid anhydrides such as methyl chlorocarbonate, ethyl chlorocarbonateic, isobutyloxycarbonyl chloride, pivaloyl chloride, and the like. Examples of the base include triethylamine, DIEPA, pyridine, N-methylmorpholine, and the like. Examples of the solvent include methanol, ethanol, isopropyl alcohol, butanol, ethylene glycol, tetrahydrofuran, chloroform, N,N-dimethylformamide, dimethyl sulfoxide, toluene, and the like. Examples of the base include triethylamine, DIEPA, pyridine, N-methylmorpholine, and the like.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an imidazolyl group, R6a1 is substituted by —(CRY11RY11′)n10-W10a—Y10b-W10b′ and further substituted by R6a2, R6a3, i.e., the following compound [O-3].
wherein each symbol is as defined above.
The formula [O-3] is obtained by an alkylation reaction of the formula [O-1] and the formula [O-2]. The alkylation reaction proceeds using a base and an alkylating agent such as halogenated alkyl and the like in an appropriate solvent at generally at 0° C. to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 30 min to 24 hr. Examples of the base include inorganic bases such as sodium hydride, potassium hydroxide, cesium carbonate, potassium carbonate, and the like, alkoxides such as potassium tert-butoxide, sodium methoxide, and the like, and the like. As the reaction promoter, for example, NaI, TBAI, or the like is used at times. Examples of the solvent include N,N-dimethylformamide, tetrahydrofuran, acetonitrile, toluene, NMP, dimethyl sulfoxide, and the like.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein W is an imidazolyl group, R6a1 is substituted by —(CRY11RY11′)n10—W10a—W10b′ (W10a is an optionally substituted aryl divalent group having 6 to 10 carbon atoms or an optionally substituted heteroaryl divalent group containing 1 to 6 atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 5 to 10 ring-constituting atoms, W10b is an optionally substituted aryl group having 6 to 10 carbon atoms or an optionally substituted heteroaryl group containing 1 to 6 same or different atoms selected from a nitrogen atom, an oxygen atom, and a sulfur atom and having 5 to 10 ring-constituting atoms) further substituted by R6a2, R6a3, i.e., the following compound [P-3].
wherein each symbol is as defined above.
The formula [P-3] is obtained by a coupling reaction of the formula [P-1] and the formula [P-2]. The reaction preferably proceeds in the presence of a palladium catalyst, a phosphine ligand, and a base in a suitable solvent at 0° C. to under heating, particularly from room temperature to the boiling point of the solvent. The reaction time and the palladium catalyst, phosphine ligand and base, and the solvent to be used are the same as those in step 2 of Synthesis method M.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein E is a bond, X is an optionally substituted aryl group or an optionally substituted heteroaryl group, i.e., the following compound [Q-3].
wherein each symbol is as defined above.
As the method for obtaining the formula [Q-3], for example, any of the following two methods is used.
The formula [Q-3] is obtained by an SnAr reaction of the formula [Q-1] and the formula [Q-2]. The reaction proceeds in the presence of a base in an appropriate solvent at generally from room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 30 min to 48 hr. Examples of the base include inorganic bases such as sodium hydride, potassium hydroxide, cesium carbonate, potassium carbonate, and the like, alkoxides such as potassium t-butoxide, sodium methoxide, and the like, and organic bases such as triethylamine, DBU, DIPEA, and the like. Examples of the solvent include tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethyl sulfoxide, NMP, toluene, xylene, acetonitrile, and the like.
The formula [Q-3] is obtained by a coupling reaction of the formula [Q-1] and the formula [Q-2]. The reaction preferably proceeds in the presence of a palladium catalyst, a phosphine ligand, and a base in a suitable solvent at 0° C. to under heating, particularly from room temperature to the boiling point of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 1 hr to 72 hr. Examples of the palladium catalyst include palladium(II) acetate, palladium (II) chloride, tris(dibenzyl ideneacetone)dipalladium(0) or chloroform adduct thereof, and the like. Examples of the phosphine ligand include triphenyl phosphine, 2-(dicyclohexylphosphino)-2′,4′,6′-triisopropyl-1,1′-biphenyl, 2-(dicyclohexylphosphino)-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl, 2-(dicyclohexylphosphino)-2,6-diisopropoxy-1,1′-biphenyl, 2-di-tert-butylphosphino-2′-4′-6′-triisopropylbiphenyl, 2-dicyclohexylphosphino-2′-6′-dimethoxybiphenyl, 2-(dicyclohexylphosphino)-2-(N,N-dimethylamino)biphenyl, tri-ortho-tolylphosphine, 2-(dicyclohexylphosphino)biphenyl, 2-(di-tert-butylphosphino)biphenyl, 2-(di-tert-butylphosphino)-1,1-binaphthyl, tri-tert-butylphosphine, tri-tert-butylphosphonium tetrafluoroborate, and the like. A reagent in which a palladium catalyst and a phosphine ligand form a complex may also be used. Examples thereof include tetrakis(triphenyl phosphine)palladium(0), 1,1-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, dichlorobis(triphenylphosphine)palladium(II), dichlorobis(tricyclohexylphosphine)palladium(II), bis(tri-tert-butylphosphine)palladium(0), chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl) (2′-amino-1,1′-biphenyl-2-yl)palladium(II), [(2-dicyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)palladium(II) methanesulfonate, (2-dicyclohexylphosphino-2,6-diisopropoxy-1,1′-biphenyl) [2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate, [(2-di-tert-butylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)palladium(II) methanesulfonate, and the like. Examples of the base include tert-butoxy sodium, potassium acetate, tripotassium phosphate, cesium carbonate, potassium carbonate, sodium hydrogen carbonate, triethylamine, diisopropyl ethylamine, dicyclohexylethylamine, potassium fluoride, cesium fluoride, and the like. Examples of the solvent include ether solvents such as tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane, and the like, alcoholic solvents such as methanol, ethanol, propanol, butanol, and the like, N,N-dimethylformamide, NMP or a mixed solvent of an organic solvent and water, and the like.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein E is —CO—, X is —CHRm—X1—X2 wherein X1 is substituted by a triazolyl group, i.e., the following compound [R1-4].
wherein PG3 is a carboxylic acid-protecting group, as the protecting group represented by PG3, for example, alkyl can be mentioned. Each symbol is as defined above.
The formula [R1-3] is obtained by a click reaction of the formula [R1-1] and the formula [R1-2]. The reaction proceeds in the presence of a suitable metal catalyst in a suitable solvent at 0° C. to the refluxing temperature of the solvent. Examples of the metal catalyst include Cu, CuSO4, Zn(OAc)2, and the like. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally 5 min to 24 hr. As a reaction promoter, SODIUM ASCORBATE and the like are used at times. As the solvent, water, methanol, ethanol, t-BuOH, dimethyl sulfoxide, and a mixed solvent thereof, and the like are used.
The formula [R1-4] is obtained by subjecting the formula [R1-3] to a basic condition or an acidic condition. The reaction proceeds using a base or acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. The reaction time and the base and the solvent to be used are the same as those in step 4 of Synthesis method C.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein E is —CO—, X is —CHRm—X1—X2 wherein X1 is substituted by a pyrazolyl group, i.e., the following compound [R-4].
wherein each symbol is as defined above.
The formula [R2-3] is obtained by an alkylation reaction of the formula [R2-1] and the formula [R2-2]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The formula [R2-4] is obtained by subjecting the formula [R2-3] to a basic condition or an acidic condition. The reaction proceeds using a base or acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. The reaction time and the base and the solvent to be used are the same as those in step 4 of Synthesis method C.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein E is —CO—, X is —CHRm—X1—X2, wherein Rm is an optionally substituted alkyl group having 1 to 10 carbon atoms, and X1 is substituted by an oxadiazolyl group, i.e., the following compound [R3-6].
wherein each symbol is as defined above.
The formula [R3-3] can be derived from the formula [R3-1] and the formula [R3-2] according to a known method (e.g., WO 2016044386).
The formula [R3-5] is obtained by an alkylation reaction of the formula [R3-3] and the formula [R3-4]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The formula [R3-6] is obtained by subjecting the formula [R3-5] to a basic condition or an acidic condition. The reaction proceeds using a base or acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. The reaction time and the base and the solvent to be used are the same as those in step 4 of Synthesis method C.
The production method described here is suitable for producing an intermediate that is a compound represented by the formula (I) wherein E is —CO—, X is —CHRm—X1—X2, wherein Rm is an optionally substituted alkyl group having 1 to 10 carbon atoms, and X1 is substituted by an isoxazolyl group, i.e., the following compound [R4-4].
wherein each symbol is as defined above.
The formula [R4-3] is obtained by an alkylation reaction of the formula [R4-1] and the formula [R4-2]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The formula [R4-4] is obtained by subjecting the formula [R4-3] to a basic condition or an acidic condition. The reaction proceeds using a base or acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. The reaction time and the base and the solvent to be used are the same as those in step 4 of Synthesis Method C.
The production method described here is suitable for producing a compound represented by the formula (I) wherein E is —CO—, X is —CHRm—X1—X2, wherein X1 is —NR1X—, X2 is —CO—, and further substituted by L-A, i.e., the following compound [S-7].
wherein each symbol is as defined above.
The formula [S-2] is obtained by deprotection of the formula [S-1] with an acid. The reaction proceeds using an appropriate acid in an appropriate solvent generally at room temperature to the refluxing temperature of the solvent. While the reaction time varies depending on the starting materials and solvents to be used and the reaction temperature, it is generally from 30 min to 24 hr. Examples of the acid include hydrochloric acid, trifluoroacetic acid, PTSA, PPTs, and the like. Examples of the solvent include 1,4-dioxane, dichloromethane, ethyl acetate, methanol, toluene, and the like.
The formula [S-4] is obtained by a condensation reaction of the formula [S-2] and the formula [S-3]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in Synthesis method N.
The formula [S-5] is obtained by deprotection of the formula [S-4] with an acid. The reaction proceeds using an appropriate acid in an appropriate solvent generally from room temperature to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in step 1 of Synthesis method N.
The formula [S-7] is obtained by a condensation reaction of the formula [S-5] and the formula [S-6]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in Synthesis method N.
The production method described here is suitable for producing a compound represented by the formula (I) wherein E is substituted by —CO—, i.e., the following compound [T-3].
wherein each symbol is as defined above.
The formula [T-3] is obtained by a condensation reaction of the formula [T-1] and the formula [T-2]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in Synthesis method N.
The production method described here is suitable for producing a compound represented by the formula (I) wherein a connection part with a chemical linker is —CONRL1—, i.e., the following compound [U-3].
wherein each symbol is as defined above.
The formula [U-3] is obtained by a condensation reaction of the formula [U-1] and the formula [U-2]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in Synthesis method N.
The production method described here is suitable for producing a compound represented by the formula (I) wherein a connection part with a chemical linker is —O—, i.e., the following compound [V-3].
wherein each symbol is as defined above.
The formula [V-3] is obtained by an alkylation reaction of the formula [V-1] and the formula [V-2]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The production method described here is suitable for producing a compound represented by the formula (I) wherein a connection part with a chemical linker is —S—, i.e., the following compound [W-3].
wherein each symbol is as defined above.
The formula [W-2] is obtained by an alkylation reaction of the formula [W-1] and the formula [V-2]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The production method described here is suitable for producing a compound represented by the formula (I) wherein a connection part with a chemical linker is —NRL1— or —NRL1CO—, i.e., the following compound [X-3] or compound [X-4].
wherein each symbol is as defined above.
The formula [X-3] is obtained by an alkylation reaction of the formula [X-1] and the formula [V-2]. The reaction proceeds using a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the base, solvent, and reaction promoter to be used are the same as those in Synthesis method O.
The formula [X-4] is obtained by a condensation reaction of the formula [X-1] and the formula [X-2]. The reaction proceeds using a condensing agent in the presence of a suitable base in a suitable solvent at 0° C. to the refluxing temperature of the solvent. The reaction time and the condensing agent, solvent, reaction promoter, and base to be used are the same as those in Synthesis method N.
The starting material compounds in the above-mentioned methods can be produced by known methods and/or in the same manner as in the methods described in Examples described later.
The introduction of protecting groups into functional groups and the removal of functional group protecting groups can be performed by reference to a known method (PROTECTIVE GROUPS in ORGANIC SYNTHESIS (Theodora W. Greene, Peter G. M. Wuts) etc.).
In addition, the compound of the present invention and intermediate compounds produced by the above-mentioned methods can be structurally converted to other compound of interest, or intermediate by the method described in Example described later and/or known methods or combination thereof.
A compound represented by the formula (I), which is produced by the aforementioned method, can be purified to any purity by a conventionally used purification means, for example, concentration, extraction, chromatography, reprecipitation, recrystallization, and the like. It can be converted to a pharmacologically acceptable salt as necessary by treating with an acid or a base etc. in a suitable solvent (water, alcohol, ether, etc.). Furthermore, the obtained compound of the present invention or a pharmacologically acceptable salt thereof can be converted to hydrate or solvate by treating with water, water-containing solvent or other solvent.
The compound and a pharmacologically acceptable salt thereof of the present invention include racemic compounds, stereoisomers, and mixture of these compounds, and includes isotope-labeled and radioactive-labeled compounds. Such isomers can be isolated by a standard separation technique including fractional crystallization and chiral column chromatography. In addition, the compound of the present invention has an asymmetric carbon atom. Therefore, it includes enantiomer and diastereomer. A diastereomer mixture can be separated into each diastereomer based on their physical/chemical differences by a method well known in the art, for example, chromatography and/or fractional crystallization. Enantiomer can be separated by chiral column chromatography or by reacting an enantiomer compound with an appropriate optically active compound to give a diastereomer mixture, separating each diastereomer and converting each diastereomer to a corresponding enantiomer. The compound of the present invention may be any of such isomers including diastereomer, enantiomer, and a mixture thereof.
The compound of the present invention has low toxicity (e.g., acute toxicity, chronic toxicity, genetic toxicity, reproductive toxicity, cardiotoxicity, carcinogenicity), and can be used as a medicament for the prophylaxis or treatment of diseases caused by dysregulation of protein activity in a mammal (e.g., mouse, rat, hamster, rabbit, cat, dog, bovine, sheep, monkey, human, etc.).
While the diseases caused by dysregulation of protein activity are not limited, for example, asthma, multiple sclerosis, cancer, cilium associated disease, cleft palate, diabetes, cardiac disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, coeliac disease, Charcot-Marie-Tooth disease, cystic fibrosis, Duchenne muscular dystrophy, haemochromatosis, haemophilia, Klinefelter's syndrome, neurofibromatosis, phenylketonuria, polycystic kidney disease, (PKD1) or 4(PKD2) Prader-Willi syndrome, sickle-cell disease, Tay-Sachs disease, Turner's syndrome;
Alzheimer's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease), anorexia nervosa, anxiety disorder, atherosclerosis, attention deficit hyperactivity disorder, autism, bipolar disorder, chronic fatigue syndrome, chronic obstructive pulmonary diseases, Crohn's disease, coronary heart disease, dementia, depression, diabetes mellitus type 1, diabetes mellitus type 2, epilepsy, Guillain-Barre syndrome, irritable bowel syndrome, lupus, metabolic syndrome, multiple sclerosis, myocardial infarction, obesity, obsessive-compulsive disorder, panic disorder, Parkinson's disease, psoriasis, rheumatoid arthritis, sarcoidosis, schizophrenia, stroke, thromboangiitis obliterans, Tourette's syndrome, Vasculitis;
aceruloplasminemia, achondrogenesis type II, achondroplasia, acrocephaly, Gaucher's disease type 2, acute intermittent porphyria, Canavan disease, adenomatous polyposis coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, adrenogenital syndrome, adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, alkaptonuria, Alexander's disease, alkaptonuric ochronosis, Alpha 1-antitrypsin deficiency, Alpha-1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis, Alström's syndrome, Alexander's disease, amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, anemia, angiokeratoma corporis diffusum, angiomatosis retinae (von Hippel-Lindau disease), Apert syndrome, arachnodactyly (Marfan's syndrome), Stickler's syndrome, arthrochalasis multiplex congenital (Ehlers-Danlos syndrome # arthrochalasia type), telangiectatic ataxia, Rett syndrome, primary pulmonary hypertension, Sandhoff's disease, neurofibromatosis type II, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever, familial, Benjamin syndrome, beta-thalassemia, bilateral acoustic neurofibromatosis (neurofibromatosis type II), Factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloom's syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome (Turner syndrome), Bourneville's disease (tuberous sclerosis), Birt-Hogg-Dube syndrome, osteoporosis (osteogenesis imperfecta), Broad Thumb-Hallux syndrome (Rubinstein-Taybi syndrome), bronze diabetes/bronzed cirrhosis (hemochromatosis), Bulbospinal muscular atrophy (Kennedy's disease), Buerger-Gruetz syndrome (lipoprotein lipase deficiency), CGD chronic granulomatosis, Campomelic dysplasia, biotinidase deficiency, cardiomyopathy (Noonan syndrome), Cri-du-Chat syndrome, CAVD (congenital absence of the was deferens), Caylor cardiofacial syndrome (CBAVD), CEP (Congenital erythropoietic porphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophy syndromes (achondroplasia), otospondylomegaepiphyseal dysplasia, Lesch-Nyhan syndrome, galactosemia, Ehlers-Danlos syndrome, thanatophoric dysplasia, Coffin-Lowry syndrome, Cockayne syndrome, familial adenomatous polyposis, Congenital erythropoietic porphyria, congenital heart disease, methemoglobinemia/congenital methemoglobinemia, achondroplasia, X-linked sideroblastic anemia, connective tissue disease, Conotruncal anomaly face syndrome, Cooley's anemia (beta-thalassemia), copper storage disease (Wilson's disease), copper transport disease (Menkes disease), hereditary coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome), Creutzfeldt-Jakob disease (prion disease), Cowden syndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria, Spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher syndrome, degenerative neurological diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disability, distal spinal muscular atrophy, type V, androgen insensitivity syndrome, diffuse globoid body sclerosis (Krabbe disease), DiGeorge syndrome, dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, dwarfism, erythropoietic protoporphyria, erythroid 5-aminolevulinate synthase deficiency, erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Friedreich's ataxia, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure-sensitive neuropathy, primary pulmonary hypertension (PPH), pancreas fibrocystic disease, fragile X syndrome, galactosemia, genetic brain disorder, giant cell hepatitis (neonatal hemochromatosis), Gronblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther's disease (congenital erythropoietic porphyria), hemochromatosis, Hallgren's syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease (von Hippel-Lindau disease), Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), hyperandrogenism, hypochondroplasia, hypochromic anemia, immune system disorders including X-linked severe combined immunodeficiency, Insley-Astley syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, renal diseases including hyperoxaluria, Klinefelter's syndrome, Kniest dysplasia, lacunar dementia, Langer-Saldino achondrogenesis, ataxia telangiectasia, Lynch syndrome, lysyl-hydroxylase deficiency, Machado-Joseph disease, metabolic disorders including Kniest dysplasia, Marfan syndrome, movement disorder, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome, multiple neurofibromatosis, Nance-Insley syndrome, Nance-Sweeney chondrodysplasia, Niemann-Pick disease, Noack syndrome (Pfeiffer syndrome), Osler-Weber-Rendu disease, Peutz-Jeghers syndrome, polycystic kidney disease, polyostotic fibrous dysplasia (McCune-Albright syndrome), Prader-Labhart-Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, progressive chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular atrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb (retinoblastoma), Recklinghausen disease (neurofibromatosis type 1), Recurrent polyserositis, retinal disorder, retinoblastoma, Rett syndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levy syndrome, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome, tuberous sclerosis, SDAT, congenital SED (spondyloepiphyseal dysplasia congenita), SED Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type (spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome, skin pigmentation disorder, Smith-Lemli-Opitz syndrome, South-African genetic porphyria (variegate porphyria), infantile-onset ascending hereditary spastic paralysis, speech and communication disorder, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, thyroid gland disease, Tomaculous neuropathy (hereditary neuropathy with liability to pressure palsies), Treacher Collins syndrome, Triplo X syndrome (triple X syndrome), trisomy 21 (Down syndrome), trisomy X, VHL syndrome (von Hippel-Lindau disease), vision impairment and blindness (Alstrom syndrome), Vrolik disease, Waardenburg syndrome, micro syndrome (Warburg Sjo Fledelius syndrome), Weissenbacher-Zweymuller syndrome, Wolf-Hirschhorn syndrome, Wolff Periodic disease, Weissenbacher-Zweymuller syndrome and xeroderma pigmentosum;
virus infectious disease such as HIV, HCV and the like, and the like can be mentioned.
In the present specification, the “prophylaxis” includes preventing the onset of a disease (all or one or more pathologies) and delaying the onset of the disease. The “prophylactically effective amount” refers to a dose of compound (I) sufficient to achieve such purpose.
In the present specification, the “treatment” includes curing a disease (all or one or more pathologies), improving the disease, and suppressing the progression of the severity of the disease. The “therapeutically effective amount” refers to a dose of compound (I) sufficient to achieve such purpose.
In practicing the present invention, the compound of the present invention (compound (I) or a pharmacologically acceptable salt thereof) can be used either in a single form, or in the form of a pharmaceutical composition containing the compound of the present invention as an active ingredient, together with a pharmaceutically acceptable carrier.
Examples of such pharmaceutical composition include tablet (including sugar-coated tablet, film-coated tablet, sublingual tablet, orally disintegrating tablet, buccal tablet, and the like), pill, powder, granule, capsule (including soft capsule and microcapsule), syrup, liquid, emulsion, suspension, controlled-release preparation (e.g., immediate-release preparation, sustained-release preparation, sustained-release microcapsule), aerosol, films (e.g., orally disintegrable films, mouth cavity mucosa pasting film), injection (e.g., subcutaneous injection, intravenous injection (e.g., bolus), intramuscular injection, intraperitoneal injection), drip transfusion, transdermal absorption type preparation, ointment, lotion, adhesive preparation, suppository (e.g., rectal suppository, vaginal suppository), pellet, nasal preparation, pulmonary preparation (inhalant), eye drop, and the like.
In the present specification, as the “pharmaceutically acceptable carrier”, various carriers conventionally used in the field of formulation technology can be used.
Specific examples of the “pharmaceutically acceptable carrier” for solid preparations include excipient (e.g., lactose, sucrose, D-mannitol, starch, cornstarch, crystalline cellulose, light anhydrous silicic acid, etc.), lubricant (e.g., magnesium stearate, talc, colloid silica, etc.), binder (e.g., crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, starch, sucrose, gelatin, methylcellulose, carboxymethylcellulose sodium, etc.), disintegrant (e.g., starch, carboxymethylcellulose, carboxymethylcellulose calcium, sodium carboxymethyl starch, L-hydroxypropylcellulose etc.), and the like.
For liquid preparations, solvent (e.g., water for injection, isotonic brine, alcohol, propylene glycol, macrogol, sesame oil, etc.), solubilizing agent (e.g., polyethylene glycol, propylene glycol, D-mannitol, benzoic acid benzyl, ethanol, triethanolamine, sodium carbonate, sodium citrate, etc.), suspending agent (e.g., surfactants such as stearyltriethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, mono stearic acid glycerol, and the like; hydrophilic polymers such as poly(vinyl alcohol), polyvinylpyrrolidone, carboxymethylcellulose sodium, methylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, and the like), isotonicity agent (e.g., glucose, D-Sorbitol, sodium chloride, glycerol, D-mannitol, etc.), buffering agent (e.g., buffers such as phosphate, citrate, and the like), soothing agent (e.g., benzyl alcohol, etc.), and the like can be used.
Where necessary, preparation additives such as antiseptic (e.g., p-hydroxybenzoic acid esters, chlorobutanol, benzyl alcohol, sorbic acid, etc.), antioxidant (e.g., sulfite, ascorbic acid, α-tocopherol, etc.), colorant, sweetening agent, and the like may be further added.
The pharmaceutical composition of the present invention can be produced by adding the compound of the present invention in a proportion of 0.01 to 99% (w/w), preferably 0.1 to 85% (w/w), based on the total amount of the preparation, though subject to change depending on the dosage form, administration method, carrier, and the like. The pharmaceutical composition can be produced by a method conventionally used in the field of formulation technology, according to its form. The pharmaceutical composition of the present invention may be formulated into a sustained-release preparation containing the active ingredient.
The compound of the present invention can be expected to have low toxicity and few side effects, and also has superior properties as a pharmaceutical product. Therefore, the compound of the present invention can be safely administered to mammals (particularly human).
In practicing the present invention, the compound of the present invention may be administered either alone or as a pharmaceutical composition orally or parenterally (e.g., intravenous, intramuscular, subcutaneous, intraorgan, intranasal, intradermal, ophthalmic, intracerebral, intrarectal, intravaginal, intraperitoneal administrations, and administration to lesion).
The dose of the compound of the present invention varies depending on the administration subject, administration route, and the age and symptoms of the administration subject, and is not particularly limited. For example, the dose of the compound of the present invention is 1 to 100 mg per dose for oral administration, and 0.1 to 1000 mg per dose for parenteral administration.
The compound (I) may also be used in the form of a prodrug thereof. A prodrug of the compound (I) means a compound which is converted to the compound (I) of the present invention with a reaction due to an enzyme, an gastric acid, etc. under the physiological condition in the living body, that is, a compound which is converted to the compound (I) of the present invention with oxidation, reduction, hydrolysis, etc. according to an enzyme; a compound which is converted to the compound (I) of the present invention by hydrolysis etc. due to gastric acid, etc. A prodrug for compound (I) may be a compound obtained by subjecting an amino group in compound (I) to an acylation, alkylation or phosphorylation (e.g., a compound obtained by subjecting an amino group in compound (I) to an eicosanoylation, alanylation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidylmethylation, pivaloyloxymethylation and tert-butylation, etc.); a compound obtained by subjecting a hydroxy group in compound (I) to an acylation, alkylation, phosphorylation or boration (e.g., a compound obtained by subjecting an hydroxy group in compound (I) to an acetylation, palmitoylation, propanoylation, pivaloylation, succinylation, fumarylation, alanylation, dimethylaminomethylcarbonylation, etc.); a compound obtained by subjecting a carboxyl group in compound (I) to an esterification or amidation (e.g., a compound obtained by subjecting a carboxyl group in compound (I) to an ethyl esterification, phenyl esterification, carboxymethyl esterification, dimethylaminomethyl esterification, pivaloyloxymethyl esterification, ethoxycarbonyloxyethyl esterification, phthalidyl esterification, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl esterification, cyclohexyloxycarbonylethyl esterification and methylamidation, etc.) and the like. These compounds can be produced from compound (I) by a method known per se.
A prodrug for compound (I) may also be one which is converted to compound (I) under physiological conditions, such as those described in IYAKUHIN no KAIHATSU (Development of Pharmaceuticals), Vol. 7, Design of Molecules, p. 163-198, Published by HIROKAWA SHOTEN (1990).
(Combined Use with Other Drugs)
As described above, the compound of the present invention has extremely low toxicity, can be used in combination with other medicaments for the prophylaxis or treatment of target diseases, and is expected to exhibit superior prophylactic and/or therapeutic effects in combination with other medicaments. Such combination therapy is also expected to lower the dose of other medicaments and reduce the side effects they have.
Such medicament that can be used in combination with the compound of the present invention (hereinafter to be abbreviated as concomitant drug) can be appropriately selected in consideration of the type of disease of the patients, the severity of its symptoms, and the like.
The administration mode of the concomitant drug is not particularly limited, and the compound of the present invention and the concomitant drug may be combined at the time of administration. For example, they can be used in the administration modes of (1) administration of a preparation containing the compound of the present invention and the concomitant drug in combination, (2) simultaneous or separate administration of two kinds of preparations of the compound of the present invention and the concomitant drug, which have been separately produced, by the same administration route, (3) simultaneous or separate administration of two kinds of preparations of the compound of the present invention and the concomitant drug, which have been separately produced, by different administration routes, and the like. A preferable form, according to the medical practice, can be appropriately selected.
A preparation containing the above-mentioned compound of the present invention and a concomitant drug in combination can be appropriately produced by those of ordinary skill in the art according to the pharmaceutical composition described above containing the compound of the present invention.
The dose of the concomitant drug can be appropriately determined based on the dose employed in clinical situations. The mixing ratio of the compound of the present invention and a concomitant drug can be appropriately determined depending on the disease and symptoms of the administration subject, administration route, the kind of the concomitant drug to be used, and the like. Generally, it can be appropriately determined based on the general clinical dose of the concomitant drug to be used and according to the actual situation in medical practice.
The present invention is explained in more detail in the following by referring to Reference Examples and Examples relating to the synthesis of compound (I), as well as Experimental Examples relating to the pharmacological activity of compound (I). However, these are only examples, and the present invention is not limited to these.
The meanings of the following notations attached to the structural formulas in the table describing the “structural formulas of Reference Example compounds” (Table 1) and the table describing the “structural formulas of Example compounds” (Table 2) described later are explained.
The notation “or 1” indicates that the carbon atom to which it is attached has been identified to have a single steric configuration, but the absolute steric configuration thereof has not been identified.
The notation “wavy line” indicates that in a compound having two or more asymmetric carbon atoms, the stereochemistry of the carbon atom to which it is attached is a mixture of R form and S form.
The notation “solid line” (bonded to asymmetric carbon atom)” indicates a racemate.
Representative abbreviations used in the description of the following Reference Examples and Examples are explained.
3,3-Dibromo-1,1,1-trifluoro-propan-2-one (5 g) was dissolved in water (61.8 mL), sodium acetate (4.4 g) was added at room temperature, and the mixture was stirred at 100° C. for 30 min. After completion of the reaction, the reaction solution was added to a mixed solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate (3 g), methanol (152 mL), and 7 M ammonia-methanol (26 mL) at 0° C., and the mixture was stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=85:15) to give the title compound (3.4 g) as a colorless powder. MS (ESI) m/z: 436.4 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (2.2 g) described in Reference Example 1 was dissolved in methanol (12 mL), 5 M sodium methoxide-methanol (3.6 mL) was added, and the mixture was stirred under microwave radiation at 100° C. for 10 min. After completion of the reaction, the reaction solution was concentrated under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (2.3 g) as a brown powder.
MS (ESI) m/z: 470.4 [M−H]−
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (1312 mg) described in Reference Example 1 in methanol (6.0 mL) was added 5 M sodium methoxide-methanol solution (2100 μL), and the mixture was stirred under microwave radiation at 100° C. for 10 min. After cooling to room temperature, water and saturated brine were added and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. To the obtained residue were added DCM (30 mL) and trifluoroacetic acid (7.5 mL), and the mixture was stirred at room temperature for 3 hr. After cooling to −15° C., 1 M sodium hydroxide aqueous solution (88 mL) was added dropwise over 8 min. After the completion of the dropwise addition, the mixture was allowed to warm to room temperature and adjusted to around pH=8 by adding sodium hydrogen carbonate (1680 mg). Ethyl acetate (30 mL) and Boc2O (1047 mg) were added, and the mixture was stirred at room temperature for 2.5 hr. After completion of the reaction, the reaction mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=76:24-55:45) to give the title compound (1033 mg) as a pale-red powder.
MS (ESI) m/z: 426.3 [M+H]+
Methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-5-carboxylate (528 mg) described in Reference Example 3 was dissolved in methanol (5 mL), 1 M sodium hydroxide aqueous solution (6.2 mL) was added and the mixture was stirred at 50° C. for 3 days. After completion of the reaction, 1 M hydrochloric acid (5 mL) was added and the reaction solution was concentrated under reduced pressure. The reaction solution was adsorbed onto PoraPak Rxn RP 60 cc, and washed with water (60 mL). Then it was eluted with methanol (60 mL), and the eluate was concentrated under reduced pressure, and azeotropically distilled with ethanol and toluene in this order to give the title compound (299 mg) as a white powder.
MS (ESI) m/z: 298.1 [M+H]+
2-[(2S,4R)-4-Hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (140 mg) described in Reference Example 4 was dissolved in N-methylpyrrolidone (940 μL), HATU (179 mg) was added at room temperature and the mixture was stirred for 20 min. A mixed solution of N-methyl-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (154 mg), diisopropyl amine (240 μL), and N-methylpyrrolidone (470 μL) was added at room temperature and the mixture was stirred for about 17 hr. At room temperature, HATU (56 mg) was added and the mixture was stirred for about 3 hr. After completion of the reaction, water and dimethyl sulfoxide were added, and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (133 mg) as a pale-yellow viscous substance. MS (ESI) m/z: 496.5 [M−H]−
tert-Butyl (2S,4R)-4-hydroxy-2-[4-[methyl-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (133 mg) described in Reference Example 5 was dissolved in methanol (1.1 mL), 4 M hydrochloric acid-dioxane (284 μL) was added at room temperature, and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the title compound (122 mg) as a pale-brown viscous substance. MS (ESI) m/z: 398.2 [M+H]+
Methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-5-carboxylate (150 mg) described in Reference Example 3 was dissolved in 1,4-dioxane (4 mL), 4 M hydrochloric acid-dioxane (6.2 mL) was added at room temperature and the mixture was stirred for 5 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and azeotropically distilled 3 times with toluene. To the obtained residue were added DMF (4 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (77 mg), diisopropyl ethylamine (183 μL), and HATU (174 mg), and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, saturated brine was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (ethyl acetate:methanol=100:0-95:5). The obtained residue was dissolved in ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (98 mg) as a pale-yellow powder. MS (ESI) m/z: 393.4 [M+H]+
Methyl 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylate (266 mg) described in Reference Example 7 was dissolved in methanol (6.8 mL), 1 M sodium hydroxide aqueous solution (6 mL) was added and the mixture was stirred at 60° C. for 19 hr. After completion of the reaction, 1 M hydrochloric acid (6 mL) was added, and the reaction solution was concentrated under reduced pressure. The reaction solution was adsorbed onto PoraPak Rxn RP 60 cc, and washed with water. It was then eluted with methanol, and the eluate was concentrated under reduced pressure to give the title compound (253 mg) as a colorless powder. MS (ESI) m/z: 379.4 [M+H]+
To 4-bromobenzaldehyde (10 g), palladium acetate (243 mg), and potassium carbonate (10.6 g) were added dimethylacetamide (18.0 mL) and 4-methylthiazole (10720 mg), and the mixture was substituted with nitrogen and stirred at 150° C. for 1.5 hr. After completion of the reaction, ethyl acetate was added, and the mixture was filtered through Celite. The filtrate was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=85:15-70:30) to give the title compound (9.2 g) as a yellow powder. MS (ESI) m/z: 204.0 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (100 mg) described in Reference Example 9 was dissolved in dichloromethane (5 mL), cyclobutylmethanamine (63 mg) was added at room temperature and stirred for 30 min. Sodium triacetoxyborohydride (150 mg) was added, and the mixture was stirred for about 1.5 hr. After completion of the reaction, 1 M sodium hydroxide aqueous solution and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by NH silica gel chromatography (hexane:ethyl acetate=95:5-70:30) to give the title compound (103 mg) as a pale-yellow oil. MS (ESI) m/z: 273.1 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (100 mg) described in Reference Example 9 was dissolved in DCM (5 mL), 2-methylpropan-1-amine (74 μL) was added and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (150 mg) was added and the mixture was stirred for 3 hr. After completion of the reaction, to the reaction mixture were added chloroform and 1 M sodium hydroxide aqueous solution, and the organic layer was separated by a Phase-separator (registered trade mark) and concentrated under reduced pressure. It was purified by NH silica gel column chromatography (hexane:ethyl acetate=95:5-70:30) to give the title compound (108 mg) as a pale-yellow oil.
MS (ESI) m/z: 261.1 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trimethoxymethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (2.3 g) described in Reference Example 2 was dissolved in dichloromethane (12 mL), TFA (48 mL) was added at 0° C., and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was extracted by adding water and 1 M sodium hydroxide aqueous solution, and a mixed solvent of ethyl acetate and methanol (methanol 10%). The organic layer was washed with saturated brine, and dried over anhydrous magnesium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by NH silica gel chromatography (ethyl acetate:methanol=95:5-50:50) The obtained residue was dissolved in DMF (10 mL), HATU (1.12 g), (2S)-2-(9H-fluorene-9-ylmethoxycarbonylamino)-3,3-dimethylbutanoic acid (1085 mg), diisopropyl ethylamine (1.5 mL) were added at room temperature and the mixture was stirred for 30 min. After completion of the reaction, it was extracted with water and ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=95:5-50:50) to give the title compound (1.5 g) as a pale-red viscous substance. MS (ESI) m/z: 661.4 [M+H]+
Methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2S)-2-(9H-fluorene-9-ylmethoxycarbonylamino)-3,3-dimethylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylate (1.5 g) described in Reference Example 12 was dissolved in methanol (7.6 mL), diethylamine (2.4 mL) was added under ice-cooling and the mixture was stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure and purified by silica gel chromatography (ethyl acetate:methanol=98:2-20:80). The obtained residue was dissolved in dichloromethane (23 mL), triethylamine (1.3 mL) and acetic anhydride (240 μL) were added at 0° C. and the mixture was stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and extracted with water and chloroform. The organic layer was washed with saturated brine, and dried over anhydrous magnesium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (ethyl acetate:methanol=100:0-95:5) to give the title compound (774 mg) as a pale-yellow viscous substance. MS (ESI) m/z: 481.3 [M+H]+
Methyl 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-[tert-butyl(dimethyl)silyl]oxypyrrolidin-2-yl]-1H-imidazole-4-carboxylate (709 mg) described in Reference Example 13 was dissolved in methanol (10 mL), 1 M sodium hydroxide aqueous solution (5.9 mL) was added while stirring at 0° C., and the mixture was heated to 50° C. and stirred overnight. After completion of the reaction, the reaction mixture was cooled to 0° C., 1 M hydrochloric acid (5.9 mL) was added, and the mixture was concentrated under reduced pressure. The reaction mixture was dissolved in water, adsorbed onto PoraPak™ Rxn RP, and washed with water (45 mL). Then the mixture was eluted with MeOH (90 mL), and concentrated under reduced pressure to give the title compound (489 mg) as a colorless powder. MS (ESI) m/z: 353.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 9 using 1-(3-bromophenyl)ethanone. MS (ESI) m/z: 218.1 [M+H]+
1-[3-(4-Methylthiazol-5-yl)phenyl]ethanone (615 mg) described in Reference Example 15 was dissolved in acetic acid (5.6 mL), trimethylphenylammonium tribromide (1596 mg) was added and the mixture was stirred with heating at 90° C. for 2 hr. After completion of the reaction, the reaction mixture was cooled to 0° C., added dropwise to saturated aqueous sodium hydrogen carbonate (70 mL) to pH10, and extracted twice with ethyl acetate (20 mL). The organic layer was washed with saturated brine, dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. To the obtained residue were added (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (785 mg) and Et3N (1.9 mL), and the mixture was stirred at room temperature for 20 hr. To the reaction mixture was added water (10 mL) and the mixture was extracted with ethyl acetate (20 mL). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (hexane:ethyl acetate=50:50-0:100) to give the title compound (489 mg) as an orange viscous substance.
MS (ESI) m/z: 447.4 [M+H]+
1-O-tert-Butyl 2-O—[2-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]-2-oxoethyl] (2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (485 mg) described in Reference Example 16 was dissolved in xylene (5.4 mL), ammonium acetate (418 mg) was added and the mixture was stirred with heating at 140° C. for 1.5 hr. The reaction mixture was allowed to cool to room temperature, saturated aqueous sodium hydrogen carbonate (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate as was concentrated under reduced pressure and purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (384 mg) as a pale-brown powder.
MS (ESI) m/z: 427.4 [M+H]+
tert-Butyl (2S,4R)-4-hydroxy-2-[5-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (380 mg) described in Reference Example 17 was dissolved in methanol (4.4 mL), 4 M hydrochloric acid-methanol (2.2 mL) was added, and the mixture was stirred with heating at 60° C. for 3 hr. The reaction mixture was concentrated under reduced pressure to give the title compound (415 mg) as a yellow powder.
MS (ESI) m/z: 327.3 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trimethoxymethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (1.2 g) described in Reference Example 2 was dissolved in DCM (25 mL), trifluoroacetic acid (6.3 mL) was added, and the mixture was stirred with heating at room temperature for 3 hr. After completion of the reaction, the mixture was azeotropically distilled with toluene, quenched with saturated aqueous sodium hydrogen carbonate, and the aqueous layer was extracted three times with ethyl acetate/10% methanol. The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in DMF (21 mL), DIPEA (1.4 mL), (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (686 mg), and HATU (1209 mg) were added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, saturated brine was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and purified by silica gel column chromatography (hexane:ethyl acetate=80:20-60:40) to give the title compound (1058 mg) as a pale-red powder.
MS (ESI) m/z: 539.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 56 using 1-(3-bromophenyl)-N-methyl-methanamine, 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole, and potassium carbonate instead of sodium carbonate. MS (ESI) m/z: 219.2 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate (4551 mg) was dissolved in 7 M ammonia-methanol (19 mL), 39% glyoxal aqueous solution (6.3 mL) was added, and the mixture was stirred with heating at room temperature for 8 hr. After quenching with saturated brine, the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (hexane:ethyl acetate=50:50) to give the title compound (2.6 g) as a colorless viscous substance.
MS (ESI) m/z: 368.3 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (170 mg) described in Reference Example 21 was dissolved in DMF (1 mL), cesium jo carbonate (450 mg) and 3-phenylbenzylbromide (174 mg) were added and the mixture was stirred with heating at 70° C. for 3 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (191 mg) as a pale-yellow oil. MS (ESI) m/z: 534.7 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (307 mg) described in Reference Example 21 was dissolved in DMF (1 mL), cesium carbonate (826 mg) and 3-chlorobenzylbromide (160 μL) were added, and the mixture was stirred with heating at 70° C. for 3.5 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (319 mg) as a pale-yellow oil. MS (ESI) m/z: 492.6, 494.6 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-chlorophenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (104 mg) described in Reference Example 23, 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (72 mg), XPhos Pd G2 (18 mg), XPhos (30 mg), sodium carbonate (90 mg), 1,4-dioxane (1 mL), and water (200 μL) were added, and the mixture was substituted with nitrogen and stirred with heating at 100° C. for 3.5 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by silica gel column chromatography (hexane:ethyl acetate=80:20-20:80) to give the title compound (38 mg) as a pale-yellow oil.
MS (ESI) m/z: 555.4 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (216 mg) described in Reference Example 21 was dissolved in DMF (1 mL), cesium carbonate (611 mg) and 1-(2-bromoethyl)-3-chlorobenzene (260 μL) were added, and the mixture was stirred with heating at 70° C. overnight. 1-(2-Bromoethyl)-3-chlorobenzene (260 μL) was added and the mixture was stirred with heating at 70° C. for 6 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (70 mg) as a pale-yellow oil. MS (ESI) m/z: 506.4/508.4 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[2-(3-chlorophenyl)ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (65 mg) described in Reference Example 25, 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (56 mg), XPhos Pd G2 (10 mg), XPhos (18 mg), sodium carbonate (56 mg), 1,4-dioxane (1 mL), and water (200 μL) were added, and the mixture was substituted with nitrogen and stirred with heating at 100° C. for 1.5 hr. It was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-20:80, chloroform:methanol=95:5) to give the title compound (74 mg) as a brown oil. MS (ESI) m/z: 569.7 [M+H]+
4-Phenylbutan-1-amine (257 mg) was dissolved in THE (5 mL), cyclopropanecarboxyaldehyde (160 μL), sodium triacetoxyborohydride (1050 mg), and acetic acid (100 μL) were added at 0° C., and the mixture was stirred at room temperature for 22 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (90 mg) as a colorless oil. MS (ESI) m/z: 204.1 [M+H]+
THE (5.0 mL) was added to 4-(4-pyridyl)butan-1-amine (204 mg), cyclopropanecarboxyaldehyde (120 μL), acetic acid (80 μL), and sodium triacetoxyborohydride (845 mg) were added at 0° C., and the mixture was stirred at room temperature for 22 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (13 mg) as a yellow oil. MS (ESI) m/z: 205.1 [M+H]+
DCM (3.0 mL) was added to 4-(4-methylthiazol-5-yl)benzaldehyde (60 mg) described in Reference Example 9, isopropylamine (38 μL) was added, and the mixture was stirred at room temperature for about 30 min. Sodium triacetoxyborohydride (90 mg) was further added, and the mixture was stirred at room temperature for about 2 hr. After completion of the reaction, to the reaction mixture were added chloroform and 1 M sodium hydroxide aqueous solution, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (hexane:ethyl acetate=95:5-70:30) to give the title compound (47 mg) as a pale-yellow oil. MS (ESI) m/z: 247.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 10 using 4-(4-methylthiazol-5-yl)benzaldehyde described in Reference Example 9 and 2,2,2-trifluoroethaneamine. MS (ESI) m/z: 287.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using 4-bromo-1H-pyrazole, ethyl 2-bromo-3-methylbutanoate, and DMSO instead of DMF.
MS (ESI) m/z: 275.1, 277.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 56 using ethyl 2-(4-bromopyrazol-1-yl)-3-methylbutanoate described in Reference Example 31 and (2-methoxy-4-pyridyl)boronic acid. MS (ESI) m/z: 304.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 8 using ethyl 2-[4-(2-methoxypyridin-4-yl)pyrazol-1-yl]-3 methylbutanoate described in Reference Example 32, and ethanol instead of methanol.
MS (ESI) m/z: 276.2 [M+H]+
2-[(2S,4R)-4-hydroxypyrrolidin-2-yl]-N-methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide hydrochloride (40 mg) described in Reference Example 6, 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (25 mg), and WSC·HCl (24 mg) were dissolved in DCM (420 μL), Et3N (35 μL) and HOBt (17 mg) were added, and the mixture was stirred at room temperature for 2.5 hr. MeOH was added and the mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (19 mg) as a white powder. MS (ESI) m/z: 577.3 [M−H]−
NMP (0.35 mL) was added to 2-[(2S,4R)-4-hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 4, HATU (26 mg) was added, and the mixture was stirred at room temperature for 30 min. Separately, a solution of N-(cyclopropylmethyl)-1-[4-(4-methylthiazol-5-yl)phenyl]methanamine (18 mg) described in Reference Example 106 in NMP (0.35 mL) and DIPEA (0.035 mL) was prepared, added to the reaction mixture, and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, water was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine/water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (26 mg) as a colorless oil. MS (ESI) m/z: 538.5 [M+H]+
tert-Butyl (2S,4R)-2-[4-[cyclopropylmethyl-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate (26 mg) described in Reference Example 35 was dissolved in 1,4-dioxane (0.4 mL), 4 M hydrochloric acid-dioxane (0.4 mL) was added at room temperature, and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and further azeotropically distilled 3 times with toluene. The obtained residue was dissolved in N,N-dimethylformamide (0.5 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (13 mg), HATU (22 mg), and diisopropyl ethylamine (34 μL) were added, and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate, and concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (19 mg) as a white powder. MS (ESI) m/z: 617.4 [M−H]−
To sodium hydride (20 mg) were added DMF (1.6 mL) and tert-butyl N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]carbamate (50 mg), and the mixture was stirred at room temperature for about 20 min. To the reaction mixture was added 2-(2-bromoethoxy)tetrahydropyran (18 mg), and the mixture was stirred at room temperature overnight. After completion of the reaction, to the reaction mixture were added chloroform and saturated ammonium chloride aqueous solution, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (18 mg) as a colorless oil. MS (ESI) m/z: 443.4 [M+H]+
DCM (20 mL) was added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (2 g) described in Reference Example 21, NBS (2130 mg) was added, and the mixture was stirred at room temperature for about 2 hr. After completion of the reaction, saturated aqueous sodium hydrogen carbonate was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-60:40) to give the title compound (2.4 g) as a colorless powder.
MS (ESI) m/z: 524.2/526.2/528.2 [M+H]+
To tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(4,5-dibromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (791 mg) described in Reference Example 38 were added ethanol (14 mL) and water (7 mL), sodium sulfite (9488 mg) was added, and the mixture was stirred at 100° C. for about 2 days. The mixture was allowed to cool to room temperature, saturated brine was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-80:20) to give the title compound (463 mg) as a white powder.
MS (ESI) m/z: 446.1/448.1 [M+H]+
To tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate (33 mg) described in Reference Example 39 were added DME (1 mL) and water (0.30 mL), potassium carbonate (31 mg), (3-methoxyphenyl)boronic acid (20 mg), and PdCl2(AmPhos)2 (5 mg) were added, and the mixture was stirred at 115° C. overnight. DME (1 mL) and water (0.30 mL) were further added, PdCl2(AmPhos)2 (5 mg) and (3-methoxyphenyl)boronic acid (17 mg) were added, and the mixture was stirred at 115° C. for 3 hr. The mixture was allowed to cool to room temperature, and filtered through Celite. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-67:33) to give the title compound (30 mg) as a colorless oil.
MS (ESI) m/z: 474.2 [M+H]+
(2S,4R)-1-tert-Butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (200 mg) was dissolved in DMF (8.6 mL), 1-amino-3-(4-methoxyphenyl)propan-2-one hydrochloride (205 mg), DIPEA (0.45 mL), and HATU (395 mg) were added at room temperature, and the mixture was stirred for about 9 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. The obtained residue was purified by silica gel chromatography (ethyl acetate:methanol=100:0-95:5). The obtained residue was dissolved in xylene (2.9 mL), ammonium acetate (113 mg) was added, and the mixture was stirred at 150° C. for about 4 hr. After completion of the reaction, sodium hydroxide aqueous solution and ethyl acetate were added, and the mixture was extracted. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (8.2 mg) as a yellow powder. MS (ESI) m/z: 374.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 40 using tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 39, and 2-[(3-methoxyphenyl)methyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane. MS (ESI) m/z: 488.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 40 using tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 39, and [2-(methylcarbamoyl)phenyl]boronic acid. MS (ESI) m/z: 501.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 23 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 1-(chloromethyl)-3-phenoxy-benzene. MS (ESI) m/z: 550.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 6 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-phenoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 44. MS (ESI) m/z: 336.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 23 using 1-(bromomethyl)-2-phenylmethoxybenzene, and tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21.
MS (ESI) m/z: 562.4 [M−H]−
The title compound was obtained by the same reaction and treatment as in Reference Example 23 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, 1-benzyloxy-3-(bromomethyl)benzene. MS (ESI) m/z: 564.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 23 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 1-(bromomethyl)-2-phenoxy-benzene. MS (ESI) m/z: 550.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 35 using 2-[(2S,4R)-4-hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 4, and N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]ethaneamine.
MS (ESI) m/z: 512.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 9 using tert-butyl N-t[(S)-1-(4-bromophenyl)ethyl]-N-methylcarbamate, 4-methylthiazole, and potassium acetate instead of potassium carbonate.
MS (ESI) m/z: 333.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 6 using tert-butyl N-methyl-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamate described in Reference Example 50 and 1,4-dioxane instead of methanol. MS (ESI) m/z: 233.2 [M+H]+
Reference Example 52: Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-[4-[methyl-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate
The title compound was obtained by the same reaction and treatment as in Reference Example 35 using 2-[(2S,4R)-4-hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 4, and (1S)—N-methyl-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethaneamine hydrochloride described in Reference Example 51. MS (ESI) m/z: 512.5 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(2-phenylmethoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (20 mg) described in Reference Example 46 in ethyl acetate (0.33 mL) was added 10% Pd/C (4 mg), and the mixture was stirred under hydrogen atmosphere at room temperature overnight. After completion of the reaction, nitrogen substitution was performed, insoluble material was filtered off through celite, washed with chloroform and the filtrate was concentrated under reduced pressure to give the title compound (15 mg) as a yellow oil. MS (ESI) m/z: 474.4 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(2-hydroxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (15 mg) described in Reference Example 53 in DMF (0.32 mL) were added cesium carbonate (32 mg) and 5-(bromomethyl)-4-methyl-thiazole hydrogen bromide (13 mg), and the mixture was stirred at 60° C. overnight. The reaction mixture was cooled to room temperature, chloroform and water were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (6.4 mg) as a brown oil. MS (ESI) m/z: 585.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 53 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-phenylmethoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 47.
MS (ESI) m/z: 474.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 54 using tert-butyl (2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[1-[(3-hydroxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 55, and 5-(bromomethyl)-4-methyl-thiazole hydrogen bromide. MS (ESI) m/z: 585.5 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (241 mg) described in Reference Example 1 was dissolved in DCM (5.5 mL), NIS (250 mg) was added, and the mixture was stirred under shading at room temperature overnight. Then, p-toluenesulfonic acid pyridinium (140 mg) was added, and the mixture was stirred under shading at room temperature for 6 hr. Sodium sulfite saturated aqueous solution was added, and the aqueous layer was extracted with dichloromethane, and the organic layer was separated by a Phase-separator (registered trade mark); and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: ethyl acetate=98:2-96:4) to give the title compound (201 mg) as a white powder. MS (ESI) m/z: 560.1 [M−H]−
The title compound was obtained by the same reaction and treatment as in Example 56 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-iodo-4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 57, and (4-methoxyphenyl)boronic acid.
MS (ESI) m/z: 542.3 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-(4-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (52 mg) described in Reference Example 58 was dissolved in 1,4-dioxane (1.3 mL), 1 M sodium hydroxide aqueous solution (0.67 mL) was added, and the mixture was stirred at room temperature for 7 hr. The mixture was stirred at 40° C. overnight, and further stirred at 50° C. for 5 hr. 1 M Sodium hydroxide aqueous solution (0.10 mL) was added, and the mixture was further stirred at 50° C. for 2 hr. 1 M Hydrochloric acid (770 μL) was added, and the reaction mixture was concentrated under reduced pressure. After azeotropic distillation with toluene, the residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (6.5 mg) as a colorless oil. MS (ESI) m/z: 404.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 54 using 2-[(2S,4R)-4-hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-5-(4-methoxyphenyl)-1H-imidazole-4-carboxylic acid described in Reference Example 59, methylamine hydrochloride, and DMF instead of DCM. MS (ESI) m/z: 417.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 40 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(4,5-dibromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 38, and (3-methoxyphenyl)boronic acid. MS (ESI) m/z: 580.3 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate (150 mg) in DCM (1518 μL) were added 1-naphthylmethanamine (100 μL) and magnesium sulfate (83 mg), and the mixture was stirred at room temperature overnight. Insoluble material was filtered off, and the filtrate was concentrated under reduced pressure. To the concentrated residue were added a solution of ammonium acetate (47 mg) in methanol (1517 μL) and 39% glyoxal aqueous solution (78 μL), and the mixture was stirred at room temperature for 7 hr. A solution of ammonium acetate (29 mg) in methanol (758 μL) and 39% glyoxal aqueous solution (39 μL) were added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and residue was purified by silica gel column chromatography. (hexane:ethyl acetate=67:33-47:53-0:100) to give the title compound (77 mg) as a pale-yellow oil.
MS (ESI) m/z: 508.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 40 using tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 39, and (4-methoxyphenyl)boronic acid.
MS (ESI) m/z: 474.2 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-(4-methoxyphenyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (27 mg) described in Reference Example 63 was dissolved in DCM (1.5 mL), trifluoroacetic acid (0.50 mL) was added, and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and then azeotropically distilled with toluene to give the title compound (34 mg) as a yellow powder. MS (ESI) m/z: 374.1 [M+H]+
To a solution of 1-bromo-3-[4-(trifluoromethyl)phenyl]propan-2-one (3765 mg) in DMF (13 mL) was added sodium diformylamide (1337 mg) under ice-cooling, and the mixture was stirred for 1 hr, after which stirred at room temperature overnight. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. To the obtained residue were added isopropanol (26 mL) and concentrated hydrochloric acid (26 mL), and the mixture was stirred at room temperature for 3 days. The reaction mixture was concentrated under reduced pressure, the obtained residue was washed with IPE, collected by filtration, and dried under reduced pressure to give the title compound (3.0 g) as a white powder. MS (ESI) m/z: 218.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 54 using (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid, 1-amino-3-[4-(trifluoromethyl)phenyl]propan-2-one hydrochloride described in Reference Example 65, and DMF instead of DCM.
MS (ESI) m/z: 429.3 [M−H]−
The title compound was obtained by the same reaction and treatment as in Reference Example 82 using tert-butyl (2S,4R)-4-hydroxy-2-[[2-oxo-3-[4-(trifluoromethyl)phenyl]propyl]carbamoyl]pyrrolidine-1-carboxylate described in Reference Example 66.
MS (ESI) m/z: 412.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 40 using tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 39, and 3-(methylaminocarbonyl)phenylboronic acid instead of (3-methoxyphenyl)boronic acid.
MS (ESI) m/z: 501.3 [M+H]+
(2S,4R)-1-tert-Butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (200 mg) was dissolved in DMF (5 mL), 1-amino-3-(4-bromophenyl)propan-2-one hydrochloride (252 mg), DIPEA (0.45 mL), and HATU (395 mg) were added, and the mixture was stirred at room temperature for about 1 hr. After completion of the reaction, water was added to the reaction solution, and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (ethyl acetate:methanol=100:0-95:5). To the obtained crude purification product were added xylene (10 mL) and ammonium acetate (338 mg), and the mixture was stirred at 150° C. for about 3 hr. After completion of the reaction, the reaction mixture was allowed to cool to room temperature, 1 M sodium hydroxide aqueous solution was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (66 mg) as a yellow oil.
MS (ESI) m/z: 422.3/424.3 [M+H]+
tert-Butyl (2S,4R)-2-[5-[(4-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate (66 mg) described in Reference Example 69 was dissolved in 1,4-dioxane (3 mL), 4 M hydrochloric acid-dioxane (1.2 mL) was added, and the mixture was stirred at room temperature for about 16 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and further azeotropically distilled with toluene. To the residue were added DMF (5.0 mL), N-Boc-L-tert-leucine (47 mg), DIPEA (0.081 mL), and HATU (83 mg), and the mixture was stirred at room temperature for about 1 hr. After completion of the reaction, 1 M sodium hydroxide aqueous solution was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (34 mg) as a pale-yellow oil. MS (ESI) m/z: 535.4/537.5 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-carbamoylpyrrolidine-1-carboxylate (976 mg) was dissolved in THF (10 mL), Lawesson reagent (916 mg) was added, and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (hexane:ethyl acetate=80:20-0:100) to give the title compound (978 mg) as a colorless powder. MS (ESI) m/z: 359.5 [M−H]−
The title compound was obtained by the same reaction and treatment as in Reference Example 21 using benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate. MS (ESI) m/z: 402.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 1-(2-bromoethyl)-4-chloro-benzene.
MS (ESI) m/z: 506.2/508.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 44 using (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid, and 1-amino-3-(2-chlorophenyl)propan-2-one hydrochloride.
MS (ESI) m/z: 395.2/397.0 [M−H]−
The title compound was obtained by the same reaction and treatment as in Reference Example 91 using tert-butyl (2S,4R)-2-[[3-(2-chlorophenyl)-2-oxopropyl]carbamoyl]-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 74. MS (ESI) m/z: 378.1/380.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 59 using tert-butyl (2S,4R)-2-[5-[(2-chlorophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 75, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid.
MS (ESI) m/z: 491.2/493.2 [M+H]+
The title compound was obtained by the same reaction and so treatment as in Reference Example 40 using tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 39, and [2-(trifluoromethyl)phenyl]boronic acid. MS (ESI) m/z: 512.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 59 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[2-(trifluoromethyl)phenyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 77, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid. MS (ESI) m/z: 511.4 [M+H]+
Under nitrogen atmosphere, 1-bromo-3-[2-(trifluoromethyl)phenyl]propan-2-one (1.0 g) was dissolved in DMF (4 mL), sodium diformylamide (379 mg) was gradually added under ice-cooling, and the mixture was stirred for 1 hr. The mixture was warmed to room temperature and stirred overnight. After completion of the reaction, the reaction was quenched by adding water, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane:ethyl acetate=90:10-67:33) to give the title compound (664 mg) as a gray powder. MS (ESI) m/z: 272.0 [M−H]−
N-Formyl-N-[2-oxo-3-[2-(trifluoromethyl)phenyl]propyl]formamide (663 mg) described in Reference Example 79 was dissolved in a mixed solution of concentrated hydrochloric acid (3.7 mL)/isopropanol (3.7 mL), and the mixture was stirred at room temperature for 2 days. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and azeotropically distilled with toluene. The obtained residue was washed with IPE in a suspension, collected by filtration and dried under reduced pressure to give the title compound (510 mg) as a gray powder. MS (ESI) m/z: 218.2 [M+H]+
(2S,4R)-1-tert-Butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (170 mg) was dissolved in DMF (7.3 mL), WSC·HCl (183 mg) and HOBt (149 mg) were added, and the mixture was stirred for about 10 min. Then, 1-amino-3-[2-(trifluoromethyl)phenyl]propan-2-one hydrochloride (205 mg) described in Reference Example 80, and Et3N (409 μL) were added, and the mixture was stirred for 4 hr. After completion of the reaction, the reaction was quenched by adding water, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate:methanol=100:0-85:15) to give the title compound (249 mg) as a brown oil.
MS (ESI) m/z: 429.3 [M−H]−
Xylene (3.4 mL) was added to tert-butyl (2S,4R)-4-hydroxy-2-[[2-oxo-3-[2-(trifluoromethyl)phenyl]propyl]carbamoyl]pyrrolidine-1-carboxylate (148 mg) described in Reference Example 31, ammonium acetate (265 mg) was added, and the mixture was stirred at 160° C. for about 1 hr in a microwave reactor. After completion of the reaction, 1 M sodium hydroxide aqueous solution was added to the reaction solution, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine/water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-80:20) to give the title compound (83 mg) as a brown oil.
MS (ESI) m/z: 412.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-hydroxy-2-[5-[[2-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 82, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid. MS (ESI) m/z: 525.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[2-(trifluoromethyl)phenyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 77, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid. MS (ESI) m/z: 525.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 25 using benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 72, and 1-bromo-4-(bromomethyl)benzene instead of 1-(2-bromoethyl)-3-chlorobenzene. MS (ESI) m/z: 570.4/572.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 56 using benzyl (2S,4R)-2-[1-[(4-bromophenyl)methyl] imidazol-2-yl]-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 85. MS (ESI) m/z: 589.6 [M+H]+
Acetic acid (3 mL) was added to benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (128 mg) described in Reference Example 86, HBr in acetic acid (0.38 mL) was added, and the mixture was stirred at room temperature for about 2 hr. After completion of the reaction, chloroform and water were added, and reversed-phase extraction was performed. To the aqueous layer was added 4 M sodium hydroxide aqueous solution, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. To the residue were added DMF (5 mL), N-Boc-L-tert-leucine (50 mg), HATU (88 mg), and DIPEA (0.075 mL), and the mixture was stirred at room temperature for about 30 min. After completion of the reaction, water was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine/water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-95:5) to give the title compound (67 mg) as a colorless oil.
MS (ESI) m/z: 554.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-2-[5-(4-bromophenyl)-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid. MS (ESI) m/z: 521.3/523.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 56 using tert-butyl N-[(2S)-1-[(2S,4R)-2-[5-(4-bromophenyl)-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 88. MS (ESI) m/z: 540.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 44 using 1-amino-3-(3-bromophenyl)propan-2-one hydrochloride and (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid instead of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid.
MS (ESI) m/z: 439.1/441.9 [M−H]−
Xylene (6.5 mL) and 1,4-dioxane (1 mL) were added to tert-butyl (2S,4R)-2-[[3-(3-bromophenyl)-2-oxopropyl]carbamoyl]-4-hydroxypyrrolidine-1-carboxylate (581 mg) described in Reference Example 90, ammonium acetate (2001 mg) was added, and the mixture was stirred at 150° C. for about 24 hr. After completion of the reaction, the reaction mixture was allowed to cool to room temperature and methanol and 1 M sodium hydroxide aqueous solution were added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (222 mg) as a yellow powder.
MS (ESI) m/z: 422.3/424.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-2-[5-[(3-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 91, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid.
MS (ESI) m/z: 535.4/537.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 56 using tert-butyl (2S,4R)-2-[5-[(3-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 91.
MS (ESI) m/z: 441.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-hydroxy-2-[5-[[3-(4-methylthiazol-5-yl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 93, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid. MS (ESI) m/z: 554.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-hydroxy-2-[5-[[4-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 67 and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid.
MS (ESI) m/z: 525.5 [M+H]+
tert-Butyl N-[(2S)-1-[(2S,4R)-2-[5-[(4-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (34 mg) described in Reference Example 70 was dissolved in 1,4-dioxane (3 mL), 4 M hydrochloric acid-dioxane (0.5 mL) was added at room temperature and the mixture was stirred for 6 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and then azeotropically distilled with toluene. The obtained residue was dissolved in dichloromethane (5 mL), triethylamine (53 μL) and acetic anhydride (12 μL) were added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, 1 M sodium hydroxide aqueous solution was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=90:10) to give the title compound (16 mg) as a colorless powder.
MS (ESI) m/z: 477.2/479.2 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[2-(4-chlorophenyl)ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (52 mg) described in Reference Example 73 was dissolved in dichloromethane (2 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and then azeotropically distilled twice with toluene. The obtained residue was dissolved in DMF (3 mL), DIPEA (89 μL), (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (29 mg), and HATU (51 mg) were added, and the mixture was stirred at room temperature for 30 min. The reaction mixture was diluted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in 1,4-dioxane (2 mL), 4 M hydrochloric acid-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hr. The reaction mixture was concentrated under reduced pressure, and the obtained residue was suspended in THF (3 mL). Triethylamine (71 μL) and acetic anhydride (29 μL) were added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (26 mg) as a pale-yellow oil. MS (ESI) m/z: 447.4/449.4 [M+H]+
DMF (2 mL), cesium carbonate (694 mg), and 3-chlorobenzylbromide (140 μL) were added to methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-5-carboxylate (299 mg) described in Reference Example 3, and the mixture was stirred at 70° C. for about 3 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-40:60) to give the title compound (191 mg) as a white powder.
MS (ESI) m/z: 550.3/552.3 [M+H]+
4-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)thiazole (102 mg), XPhos Pd G2 (25 mg), RuPhos (42 mg), sodium carbonate (130 mg), 1,4-dioxane (2 mL), and water (400 μL) were added to methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1-[(3-chlorophenyl)methyl]imidazole-4-carboxylate (160 mg) described in Reference Example 98, and the mixture was stirred under nitrogen atmosphere at 100° C. for about 2.5 hr. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-20:80) to give the title compound (185 mg) as a pale-yellow oil. MS (ESI) m/z: 613.7 [M+H]+
DMF (2 mL), cesium carbonate (283 mg), and methyl 2-bromoacetate (60 μL) were added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (231 mg) described in Reference Example 21, and the mixture was stirred at room temperature for about 20 hr. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-40:60) to give the title compound (220 mg) as a pale-yellow oil. MS (ESI) m/z: 440.5 [M+H]+
4-Carboxyphenylboronic acid (22 mg), PdCl2(AmPhos)2 (4.8 mg), potassium carbonate (28 mg), DME (0.67 mL), and water (0.22 mL) were added to tert-butyl (2S,4R)-2-(5-bromo-1H-imidazol-2-yl)-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate (30 mg) described in Reference Example 39, and the mixture was stirred under nitrogen atmosphere and heating under reflux conditions for about 3 hr. After cooling to room temperature, 4-carboxyphenylboronic acid (22 mg), PdCl2(AmPhos)2 (4.8 mg), potassium carbonate (28 mg), DME (1.3 mL), and water (0.4 mL) were added, and the mixture was stirred under nitrogen atmosphere and heating under reflux conditions for about 18 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (28 mg) as a yellow oil. MS (ESI) m/z: 488.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using methyl 2-[(2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-5-carboxylate described in Reference Example 3 and 4-methoxybenzylchloride instead of 3-phenylbenzylbromide. MS (ESI) m/z: 546.5 [M+H]+
DCM (3 mL) was added to methyl 2-[(2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1-[(4-methoxyphenyl)methyl]imidazole-4-carboxylate (195 mg) described in Reference Example 102, DIBAL-H (1.0 mol/L toluene solution, 1.5 mL) was added at 0° C., and the mixture was stirred for about 2 hr. Ethyl acetate and Rochelle salt were added and stirred at 0° C. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform:methanol=99:1-80:20) to give the title compound (132 mg) as a white powder.
MS (ESI) m/z: 518.6 [M+H]+
THE (1 mL) was added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(hydroxymethyl)-1-[(4-methoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (56 mg) described in Reference Example 103, benzyl bromide (25 μL), sodium hydride (8 mg), and TBAI (6 mg) were added at 0° C., and the mixture was stirred while gradually allowing to warm to room temperature, and the mixture was stirred at room temperature for about 8 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (64 mg) as a pale-yellow oil.
MS (ESI) m/z: 608.7 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(4-methoxyphenyl)methyl]-4-(phenylmethoxymethyl)imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 104, and (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid instead of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid.
MS (ESI) m/z: 607.4 [M+H]+
To 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (65 mg), 1-(4-bromophenyl)-N-(cyclopropylmethyl)methanamine (57 mg), PdCl2(PPh3)2 (20 mg), potassium carbonate (67 mg), and Pd(dppf)Cl2·CH2Cl2 (20 mg) were added water (0.48 mL) and 1,4-dioxane (1.2 mL), and the mixture was stirred under nitrogen atmosphere at 100° C. overnight. To the reaction mixture were added anhydrous sodium sulfate and chloroform, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (18 mg) as a colorless oil.
MS (ESI) m/z: 259.1 [M+H]+
To a solution of benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(4,5-diiodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (324 mg) described in Reference Example 147 in water (4 mL) and ethanol (4 mL) was added sodium sulfite (688 mg), and the mixture was stirred with heating under reflux for 2 days. After cooling to room temperature, saturated brine was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (120 mg) as a colorless viscous substance. MS (ESI) m/z: 528.3 [M+H]+
To a solution of benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 107 (120 mg) in THF (5 mL) was added 1-chloro-4-ethynylbenzene (40 mg), DIPEA (0.24 mL) deaerate. PdCl2(PPh3)2 (8.0 mg), copper (I) iodide (6.5 mg), under nitrogen atmosphere 80° C. for 2 hr stirred. After cooling to room temperature, anhydrous sodium carbonate and silica gel, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=100:0-90:10) to give the title compound (120 mg) as a yellow oil.
MS (ESI) m/z: 536.4/538.4 [M+H]+
To a solution of benzyl (2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[5-[2-(4-chlorophenyl) ethynyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (120 mg) described in Reference Example 108 in methanol (5 mL) were added 4 M hydrochloric acid-dioxane (1.1 mL) and 10% Pd/C (24 mg), and the mixture was stirred under hydrogen atmosphere at room temperature overnight. After nitrogen substitution, the mixture was filtered, and the filtrate was concentrated under reduced pressure. To the obtained residue were added acetic acid (3 mL) and 33% hydrogen bromide-acetic acid solution (1 mL), and the mixture was stirred at room temperature for 6 hr. The reaction mixture was concentrated under reduced pressure, and azeotropically distilled 3 times with toluene. To the obtained residue were added DMF (5 mL), (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (155 mg), DIPEA (387 μL), and HATU (340 mg), and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, saturated brine was added, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) and concentrated under reduced pressure. To the obtained residue were added methanol (10 mL) and 1 M sodium hydroxide aqueous solution (10 mL), and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove methanol, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (40 mg) as a pale-yellow oil.
MS (ESI) m/z: 505.4/507.4 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (500 mg) described in Reference Example 21 in DCM (9 mL) was added NIS (612 mg), and the mixture was stirred at room temperature for 2 hr. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-40:60) to give the title compound (739 mg) as a colorless powder.
MS (ESI) m/z: 620.2 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 112 (369 mg) in THF (5 mL) were added 1-chloro-3-ethynyl-benzene (133 mg) and DIPEA (0.78 mL), and the mixture was deaerated. PdCl2(PPh3)2 (26 mg) and copper (I) iodide (21 mg) were added, and the mixture was stirred under nitrogen atmosphere at 80° C. for 2 hr. After cooling to room temperature, anhydrous sodium carbonate and silica gel were added, and the mixture was filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-50:50) to give the title compound (298 mg) as a pale-yellow oil. MS (ESI) m/z: 502.4/504.4 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(4,5-diiodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (739 mg) described in Reference Example 110 in water (10 mL) and ethanol (30 mL) was added sodium sulfite (7519 mg), and the mixture was stirred with heating under reflux overnight. After cooling to room temperature, saturated brine was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-85:15) to give the title compound (369 mg) as a colorless viscous substance. MS (ESI) m/z: 494.3 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[2-(3-chlorophenyl)ethynyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (145 mg) described in Reference Example 111 in methanol (5 mL) were added 4 M hydrochloric acid-dioxane (2.2 mL) and 10% Pd/C (174 mg), and the mixture was stirred under hydrogen atmosphere at room temperature overnight. After nitrogen substitution, the mixture was filtered, and the filtrate was concentrated under reduced pressure. To the obtained residue were added (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (100 mg), DMF (5 mL), DIPEA (0.30 mL), and HATU (176 mg), and the mixture was stirred at room temperature for 2 hr. After completion of the reaction, saturated brine was added, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (74 mg) as a colorless oil.
MS (ESI) m/z: 505.4/507.4 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (1256 mg) described in Reference Example 21, 1-bromo-3-(bromomethyl)benzene (1281 mg), cesium carbonate (3349 mg), and DMF (4.3 mL) were mixed and stirred at 70° C. for 1.5 hr. After cooling to room temperature, acetic acid (195 μL) was added and the mixture was stirred for 15 min. Saturated ammonium chloride aqueous solution was added, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=32:68-11:89) to give the title compound (1358 mg) as a yellow oil. MS (ESI) m/z: 536.4/538.41 [M+H]+
To a solution of methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazole-5-carboxylate (50 mg) described in Reference Example 3 in DCM (1176 μL) was added NIS (53 mg), and the mixture was stirred under shading at room temperature overnight. After completion of the reaction, sodium sulfite and saturated aqueous sodium hydrogen carbonate solution were added, and the mixture was extracted with chloroform. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-70:30) to give the title compound (56 mg) as a colorless viscous substance. MS (ESI) m/z: 552.4 [M+H]+
DME (1 mL) was added to methyl 2-[(2S,4R)-1-tert-butoxycarbonyl-4-[tert-butyl(dimethyl)silyl]oxy-pyrrolidin-2-yl]-4-iodo-1H-imidazole-5-carboxylate (56 mg) described in Reference Example 115, 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (14 μL), Pd(dppf)Cl2·CH2Cl2 (8.9 mg), and tripotassium phosphate (67 mg), and the mixture was stirred under nitrogen atmosphere at 80° C. overnight. The reaction mixture was concentrated under reduced pressure, to the concentrated residue were added XPhos Pd G2 (9.5 mg), 1,4-dioxane (1 mL), water (100 μL), and 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (14 μL), and the mixture was stirred under nitrogen atmosphere at 80° C. for 6 hr. XPhos Pd G2 (13 mg) and 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (14 μL) were added, and the mixture was stirred under nitrogen atmosphere at 80° C. for 3 days. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=87:13-66:34) to give the title compound (26 mg) as a white powder. MS (ESI) m/z: 440.5 [M+H]+
To a solution of methyl 2-[(2S,4R)-1-tert-butoxycarbonyl-4-[tert-butyl(dimethyl)silyl]oxy-pyrrolidin-2-yl]-4-methyl-1H-imidazole-5-carboxylate (27 mg) described in Reference Example 116 in methanol (610 μL) was added 1 M sodium hydroxide aqueous solution (244 μL), and the mixture was stirred at 50° C. overnight. 1 M Sodium hydroxide aqueous solution (244 μL) was added and the mixture was further stirred for 4 hr and cooled to room temperature. 1 M Hydrochloric acid (488 μL) and ethanol were added, and the mixture was concentrated under reduced pressure, and azeotropically distilled with ethanol and toluene. To the obtained residue were added N-methyl-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (36 mg), HATU (32 mg), DMF (610 μL), and DIPEA (31 μL), and the mixture was stirred at room temperature overnight. HATU (20 mg) was added, and the mixture was stirred at room temperature overnight, and the reaction was discontinued by adding water (100 μL). The reaction mixture was diluted with DMSO and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (14 mg) as a pale-yellow powder. MS (ESI) m/z: 512.3 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate (301 mg) in DCM (3.0 mL) were added magnesium sulfate (170 mg) and (1R)-1-(1-naphthyl)ethaneamine (219 μL), and the mixture was stirred at room temperature overnight. The mixture was filtered and concentrated under reduced pressure. To the concentrated residue was added a solution of ammonium acetate (118 mg) in methanol (3.0 mL), 39% glyoxal aqueous solution (160 μL) was added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the concentrated residue was purified by silica gel column chromatography (hexane:ethyl acetate=85:15-64:36) to give the title compound (138 mg) as an orange oil. MS (ESI) m/z: 522.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 118 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate, [2-[4-(trifluoromethyl)phenyl]-4-pyridyl]methanamine hydrochloride described in Reference Example 148, and triethylamine. MS (ESI) m/z: 603.6 [M+H]+
DMF (2 mL), cesium carbonate (326 mg), and 1-bromo-4-(2-bromoethyl)benzene (300 μL) were added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (236 mg) described in Reference Example 21, and the mixture was stirred at 70° C. for about 18 hr. Furthermore, 1-bromo-4-(2-bromoethyl)benzene (300 μL) was added and the mixture was stirred for 1.5 hr. Again, 1-bromo-4-(2-bromoethyl)benzene (300 μL) was added and the mixture was stirred for 1 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (174 mg) as an orange oil.
MS (ESI) m/z: 550.7/552.7 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 3-(chloromethyl)-5-(4-fluorophenyl)pyridine salt acid.
MS (ESI) m/z: 553.3 [M+H]+
1,4-Dioxane (30 mL) and 4 M hydrochloric acid-dioxane (22 mL) were added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (1.1 g) described in Reference Example 21 at room temperature, and the mixture was stirred for about 4 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene to give the title compound (716 mg) as a pale-brown powder. MS (ESI) m/z: 153.9 [M+H]+
(2S,4R)-1-tert-Butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (151 mg) was dissolved in DMF (2 mL), benzene-1,2-diamine (78 mg), HATU (287 mg), and DIPEA (400 μL) were added at room temperature, and the mixture was stirred for 2 hr. After completion of the reaction, chloroform and water were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. To the obtained residue was added acetic acid (2 mL), and the mixture was stirred at 60° C. for 4 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (143 mg) as a white powder.
MS (ESI) m/z: 304.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 24 using tert-butyl (2S,4R)-2-[1-[2-(4-bromophenyl)ethyl]imidazol-2-yl]-4-[tert-butyl(dimethyl)silyl]oxy-pyrrolidine-1-carboxylate described in Reference Example 120, and 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole. MS (ESI) m/z: 569.7 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 5-(chloromethyl)-3-phenyl-1,2,4-oxadiazole.
MS (ESI) m/z: 526.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 22 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate described in Reference Example 21, and 4-(chloromethyl)-3,5-dimethyl-1-phenyl-pyrazole.
MS (ESI) m/z: 552.6 [M+H]+
tert-Butyl (2S,4R)-2-[1-[(3-bromophenyl)methyl]imidazol-2-yl]-4-[tert-butyl (dimethyl)silyl]oxy-pyrrolidine-1-carboxylate (1358 mg) described in Reference Example 114 was dissolved in DCM (17 mL), trifluoroacetic acid (7.7 mL) was added at room temperature, and the mixture was stirred for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene. To the obtained residue were added ethyl acetate and saturated aqueous sodium hydrogen carbonate, and sodium chloride was added. The organic layer was extracted and concentrated under reduced pressure. The obtained residue was dissolved in DMF (13 mL), (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (761 mg), DIPEA (1.8 mL), and HATU (1443 mg) were added at room temperature, and the mixture was stirred overnight. After completion of the reaction, ethyl acetate and water were added, and the organic layer was extracted and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=90:10-50:50) to give the title compound (1.4 g) as a pale-yellow powder. MS (ESI) m/z: 649.3/651.3 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-2-[1-[(3-bromophenyl)methyl]imidazol-2-yl]-4-[tert-butyl(dimethyl)silyl]oxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (315 mg) described in Reference Example 127, 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (131 mg), Pd(dppf)Cl2·CH2Cl2 (40 mg), and sodium carbonate (154 mg) were dissolved in 1,4-dioxane (4 mL) and water (1 mL). After nitrogen substitution, the mixture was stirred at 90° C. for 2 hr. 4-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (131 mg), and Pd(dppf)Cl2-CH2Cl2 (40 mg) were added. After nitrogen substitution again, the mixture was stirred at 90° C. for 1 hr. After completion of the reaction, anhydrous sodium sulfate was added, and the reaction solution was filtered through celite. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-30:70). The obtained residue was dissolved in 1,4-dioxane (4.8 mL), 4 M hydrochloric acid-dioxane (5 mL) was added at room temperature, and the mixture was stirred for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and azeotropically distilled with toluene to give the title compound (205 mg) as a pale-yellow powder.
MS (ESI) m/z: 454.4 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[2-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (244 mg) described in Reference Example 26 was dissolved in DCM (2.9 mL), trifluoroacetic acid (1.3 mL) was added under ice-cooling, and the mixture was stirred at room temperature for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene. To the obtained residue were added ethyl acetate and saturated aqueous sodium hydrogen carbonate, and sodium chloride was added. The organic layer was extracted and concentrated under reduced pressure. The obtained residue was dissolved in DMF (2.1 mL), (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (129 mg), DIPEA (0.30 mL), and HATU (245 mg) were added at room temperature, and the mixture was stirred overnight. After completion of the reaction, ethyl acetate and water were added, and the organic layer was extracted and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-40:60) to give the title compound (257 mg) as a colorless oil.
MS (ESI) m/z: 682.7 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 18 using tert-butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[1-[2-[3-(4-methylthiazol-5-yl)phenyl]ethyl]imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate described in Reference Example 129 and 1,4-dioxane instead of methanol.
MS (ESI) m/z: 468.5 [M+H]+
Methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2S)-3,3-dimethyl-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylate (1000 mg) described in Reference Example 19 was dissolved in 1,4-dioxane (9 mL), 4 M hydrochloric acid-dioxane (14 mL) was added at room temperature, and the mixture was stirred for 3 hr. After completion of the reaction, the reaction mixture was washed with diisopropyl ether in a suspension, and the solid was collected by filtration and dried under reduced pressure. The obtained residue was dissolved in DMF (9 mL), 1-fluorocyclopropanecarboxylic acid (213 mg), DIPEA (0.96 mL), and HATU (776 mg) were added at room temperature and the mixture was stirred for 15 hr. After completion of the reaction, 2 M ammonia-methanol was added, and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (734 mg) as a colorless powder. MS (ESI) m/z: 411.4 [M+H]+
DCM (2.8 mL) was added to methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2S)-3,3-dimethyl-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylate (152 mg) described in Reference Example 19 at an outer temperature of 0° C., DIBAL-H (1.0 mol/L toluene solution) (0.83 mL) was added, and the mixture was stirred under nitrogen atmosphere at the same temperature for 30 min, and stirred at room temperature overnight. After cooling to 0° C., the reaction mixture was diluted with diethyl ether, water (34 uL), 15% sodium hydroxide aqueous solution (34 uL), and water (85 uL) were added, and the mixture was stirred at room temperature for 30 min. Magnesium sulfate was added and the mixture was further stirred for 15 min. Insoluble material was removed from the reaction mixture by filtration through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography to give the title compound (60 mg) as a colorless oil. MS (ESI) m/z: 511.3 [M+H]+
Methyl 2-[(2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylate (734 mg) described in Reference Example 131, 1 M sodium hydroxide aqueous solution (5.7 mL), and methanol (9.5 mL) were added, and the mixture was stirred at an outer temperature of 50° C. for hr. 1 M sodium hydroxide aqueous solution (5.7 mL) was added, and the mixture was further stirred at the same temperature for 21 hr. 1 M hydrochloric acid (11 mL) was added to adjust the pH to about 3, and the mixture was concentrated under reduced pressure. The concentrated residue was washed with water in a suspension and collected by filtration. The filtrate was concentrated, the residue was dissolved in water, applied to PoraPak™ Rxn Rp (conditioned with MeOH: 45 mL, then conditioned with water: 45 mL), washed with water: 45 mL, eluted with methanol: 90 mL, and mixed with the residue obtained earlier. Three times of azeotropic distillation with toluene gave the title compound (624 mg) as a pale-brown powder.
MS (ESI) m/z: 395.3 [M−H]−
4-(4-Methylthiazol-5-yl)benzaldehyde (100 mg) described in Reference Example 9, DCM (5 mL), and cyclopropylamine. (0.051 mL) were added, and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (150 mg) was added, and the mixture was further stirred at the same temperature overnight. To the reaction mixture were added chloroform and 1 M sodium hydroxide aqueous solution, and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (hexane:ethyl acetate=95:5-72:28) to give the title compound (61 mg) as a pale-yellow oil.
MS (ESI) m/z: 245.1 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (60 mg) described in Reference Example 9, DCM (3 mL), and tetrahydrofuran-3-amine (0.038 mL) were added, and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (90 mg) was added, and the mixture was stirred at the same temperature for 1 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (58 mg) as a pale-yellow oil.
MS (ESI) m/z: 275.2 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (60 mg) described in Reference Example 9, DCM (3 mL), and tetrahydropyran-4-amine (0.046 mL) were added, and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (90 mg) was added, and the mixture was stirred at the same temperature for 2 hr was stirred. acetic acid (0.017 mL), and the mixture was further stirred at the same temperature for 3.5 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (11 mg) as a pale-yellow oil.
MS (ESI) m/z: 289.3 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (60 mg) described in Reference Example 9, DCM (3 mL), and cyclobutanamine (0.038 mL) were added, and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (90 mg) was added, and the mixture was further stirred at the same temperature for 2 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (53 mg) as a pale-yellow oil.
MS (ESI) m/z: 259.0 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (80 mg) described in Reference Example 9, DCM (4 mL), and cyclopentanamine (0.058 mL) were added, and the mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (125 mg) was added, and the mixture was further stirred at the same temperature for 2 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (75 mg) as a colorless oil.
MS (ESI) m/z: 273.1 [M+H]+
4-(4-Methylthiazol-5-yl)benzaldehyde (80 mg) described in Reference Example 9, DCM (4 mL), and cyclohexanamine (0.067 mL) were added, and the mixture was stirred at room temperature for 1 hr. Sodium triacetoxyborohydride (125 mg) was added, and the mixture was further stirred at the same temperature for 2 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (81 mg) as a colorless oil.
MS (ESI) m/z: 287.1 [M+H]+
To a solution of 4-phenylbutanoic acid (1.6 g) in THE (20 mL) were added WSC·HCl (1.9 g) and N-ethylmorpholine (2.2 g) at room temperature, and the mixture was stirred at room temperature for 1 hr. tert-Butyl N-(2-aminoethyl)carbamate (1.6 mL) was added, and the mixture was further stirred at the same temperature for 20 hr. To the reaction solution were added saturated aqueous sodium hydrogen carbonate and water and the mixture was stirred and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (2.1 g) as a white powder.
MS (ESI) m/z: 207.1 [M+H-Boc]+
To a solution of tert-butyl N-[2-(4-phenylbutanoylamino)ethyl]carbamate (1.0 g) described in Reference Example 140 and THF (10 mL) was added dropwise a borane-THF complex (11 mL) over 15 min, and the mixture was stirred at room temperature for 1 hr. To the reaction mixture was added methanol 15 mL, and the mixture was stirred and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10), and further purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (590 mg) as a colorless oil.
MS (ESI) m/z: 293.4 [M+H]+
To a solution of 2-[(2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (300 mg) described in Reference Example 133 in DMF (6.0 mL) were added DIPEA (420 μL), HATU (300 mg), and tert-butyl N-[2-(4-phenylbutylamino)ethyl]carbamate (225 mg) described in Reference Example 141 at room temperature, and the mixture was stirred at room temperature for 1 hr. The reaction mixture was diluted with DMSO and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (400 mg) as a white powder.
MS (ESI) m/z: 671.4 [M+H]+
To a solution of tert-butyl N-[2-[[2-[(2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carbonyl]-(4-phenylbutyl)amino]ethyl]carbamate (400 mg) described in Reference Example 142 in 1,4-dioxane (3.0 mL) was added 4 M hydrochloric acid-methanol (1.5 mL) at room temperature, and the mixture was stirred at room temperature for 3 hr. The reaction mixture was concentrated, azeotropically distilled with diethyl ether, and dried under reduced pressure overnight to give the title compound (372 mg) as a white powder.
MS (ESI) m/z: 571.3 [M+H]+
To a solution of 4-phenylbutanal (580 mg) and tert-butyl N-(2-aminoethyl)-N-methyl-carbamate (0.77 mL) in methanol (20 mL) were added DIPEA (0.75 mL), acetic acid (0.45 mL), and borane; 2-methylpyridine (628 mg) at room temperature, and the mixture was stirred at room temperature for 20 hr. The reaction mixture was diluted with methanol, purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile), and further purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (125 mg) as a colorless oil.
MS (ESI) m/z: 307.2 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(hydroxymethyl)-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (60 mg) described in Reference Example 132, DCM (1.2 mL), and manganese dioxide (51 mg) were added, and the mixture was stirred at room temperature for 2 hr. Manganese dioxide (51 mg) was further added, and the mixture was stirred at room temperature for 15 hr. Insoluble material was removed from the reaction mixture by filtration through diatomaceous earth, and the filtrate was concentrated under reduced pressure. To the concentrated residue were added N-(cyclopropylmethyl)-4-phenyl-butan-1-amine (36 mg) described in Reference Example 27, methanol (1.2 mL), and decaborane (7.2 mg), and the mixture was stirred at room temperature for 3 hr. N-(cyclopropylmethyl)-4-phenyl-butan-1-amine (36 mg) described in Reference Example 27 and decaborane (7.2 mg) were added, and the mixture was further stirred for 1 hr. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (70 mg) as a colorless oil.
MS (ESI) m/z: 696.4 [M+H]+
2-[(2S,4R)-1-[(2S)-2-(tert-Butoxycarbonylamino)-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (208 mg) described in Reference Example 149 was dissolved in NMP (2.5 mL), HATU (212 mg), and DIPEA (0.26 mL) were added at room temperature, and the mixture was stirred for 5 min. Then N-(cyclopropylmethyl)-4-phenyl-1-butan-1-amine (124 mg) described in Reference Example 27 was added and the mixture was stirred for 2 hr. To the reaction mixture were added water and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=50:50-0:100, ethyl acetate:methanol=9:1). The residue was dissolved in 1,4-dioxane (2.1 mL), 4 M hydrochloric acid-dioxane (1.57 mL) was added at room temperature and the mixture was stirred for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and diluted with IPE. The precipitate was collected by filtration from the reaction mixture, washed with IPE and dried. The obtained residue was dissolved in dichloromethane (0.35 mL), acetic anhydride (4 μL) and triethylamine (20 μL) were added, and the mixture was stirred at room temperature for 2 hr. To the reaction mixture was added saturated aqueous sodium hydrogen carbonate, chloroform and water were added, and the mixture was stirred. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10), and further purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (12 mg) as a colorless powder. MS (ESI) m/z: 538.6 [M+H]+
To a solution of benzyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (200 mg) described in Reference Example 72 in DCM (5 mL) was added NIS (247 mg), and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane:ethyl acetate=80:20-50:50) to give the title compound (324 mg) as a colorless oil.
MS (ESI) m/z: 654.3 [M+H]+
To 2-[4-(trifluoromethyl)phenyl]pyridine-4-carbonitrile (11 g) were added methanol (230 mL), 10% Pd/C (1.1 g), and 4 M hydrochloric acid-dioxane (23 mL), and the mixture was stirred under hydrogen atmosphere at room temperature overnight. After celite filtration, the filtrate was diluted with ethyl acetate, and the precipitated powder was collected by filtration and dried to give the title compound (14 g) as a brown powder.
MS (ESI) m/z: 253.1 [M+H]+
Under ice-cooling, methanol (10 mL) and 1 M sodium hydroxide aqueous solution (6.8 mL) were added to methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2S)-3,3-dimethyl-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylate (920 mg) described in Reference Example 19, and the mixture was stirred at 50° C. overnight. After completion of the reaction, the mixture was ice-cooled, adjusted to pH=4 by adding 1 M hydrochloric acid, and concentrated under reduced pressure. The obtained residue was washed with water and hexane:ethyl acetate=1:1 and dried under reduced pressure to give (625 mg) as a pale-yellow powder.
MS (ESI) m/z: 411.3 [M+H]+
2-Naphthylmethanamine (358 mg) was dissolved in a mixed solvent of ethyl acetate (8.4 mL) and THE (8.4 mL), tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-formylpyrrolidine-1-carboxylate (501 mg) and magnesium sulfate (279 mg) were added, and the mixture was stirred at room temperature for 2 days. Insoluble material was filtered off through celite, and the filtrate was concentrated under reduced pressure. To the obtained residue were added methanol (10 mL), ammonium acetate (212 mg), and 39% glyoxal aqueous solution (260 μL), and the mixture was stirred at room temperature for 4 days. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane:ethyl acetate=55:45-34:66-0:100). The obtained crude product was purified again by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (18 mg) as a pale-yellow oil.
MS (ESI) m/z: 508.5 [M+H]+
To a solution of tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-(2-naphthylmethyl)imidazol-2-yl]pyrrolidine-1-carboxylate (18 mg) described in Reference Example 150 in methanol (350 μL) was added 4 M hydrochloric acid-dioxane (90 μL), and the mixture was stirred at room temperature for 3 weeks. To the reaction mixture was added DIPEA (200 μL), and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (8.4 mg) as a white powder.
MS (ESI) m/z: 294.1 [M+H]+
tert-Butyl (2S)-2-amino-3-methylbutanoate hydrochloride (62 mg) and DMAP (102 mg) were dissolved in acetonitrile (1 mL), 2-azido-1,3-dimethylimidazolinium hexafluorophosphate (102 mg) was added and the mixture was stirred at 30° C. for 2.5 hr. Then, 5-ethynyl-3-methyl-isoxazole (65 mg), Copper(II) Sulfate (II) (20 mg), and sodium ascorbate (75 mg) were added at room temperature and the mixture was stirred for 19 hr. After completion of the reaction, the reaction solution was purified by silica gel column chromatography (hexane:ethyl acetate=95:5-60:40) to give the title compound (51 mg) as a white powder.
MS (ESI) m/z: 307.3 [M+H]+
tert-Butyl (2S)-3-methyl-2-[4-(3-methylisoxazol-5-yl)triazol-1-yl]butanoate (48 mg) described in Reference Example 152 was dissolved in DCM (1 mL), trifluoroacetic acid (1 mL) was added at room temperature, and the mixture was stirred for 22 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the title compound (60 mg) as an orange oil.
MS (ESI) m/z: 251.2 [M+H]+
tert-Butyl (2S,3S)-2-amino-3-methyl-pentanoate hydrochloride (106 mg) and DMAP (174 mg) were dissolved in acetonitrile (3 mL), 2-azido-1,3-dimethylimidazolinium hexafluorophosphate (153 mg) was added and the mixture was stirred at 30° C. for 2.5 hr. Then, ethoxyacetylene (150 μL), Copper(II) Sulfate (II) (31 mg), sodium ascorbate (102 mg), and water (1 mL) were added at room temperature and the mixture was stirred for 24 hr. After completion of the reaction, the reaction solution was purified by silica gel column chromatography (hexane:ethyl acetate=95:5-70:30) to give the title compound (67 mg) as a colorless oil.
MS (ESI) m/z: 284.3 [M+H]+
tert-Butyl (2S,3S)-2-(4-ethoxytriazol-1-yl)-3-methyl-pentanoate (65 mg) described in Reference Example 154 was dissolved in DCM (1 mL), trifluoroacetic acid (1 mL) was added at room temperature and the mixture was stirred for 22 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the title compound (78 mg) as a pale-yellow oil. MS (ESI) m/z: 228.1 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(hydroxymethyl)-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (75 mg) described in Reference Example 132 was dissolved in DCM (1.5 mL), manganese dioxide (127 mg) was added at room temperature, and the mixture was stirred for 15 hr. After completion of the reaction, the reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in methanol (1.5 mL), decaborane (8.9 mg) and 4-phenylbutylamine (65 mg) were added at room temperature and the mixture was stirred for 2 days. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by basic silica gel column chromatography (chloroform:methanol=90:10) to give a crude product. The obtained crude product was dissolved in DMF (1.5 mL), HATU (222 mg), DIPEA (127 μL), and cyclopropanecarboxylic acid (46 μL) were added at room temperature and the mixture was stirred for 3 hr. After completion of the reaction, the mixture was extracted with water and ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (65 mg) as a yellow oil. MS (ESI) m/z: 710.3 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(hydroxymethyl)-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (100 mg) described in Reference Example 132 was dissolved in DCM (2.0 mL), manganese dioxide (170 mg) was added at room temperature and the mixture was stirred for 15 hr. After completion of the reaction, the reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in methanol (2.0 mL), decaborane (12 mg) and cyclopropylmethylamine (42 mg) were added at room temperature and the mixture was stirred for 18 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by basic silica gel column chromatography (chloroform:methanol=90:10) to give a crude product. The obtained crude product was dissolved in DMF (2.0 mL), HATU (297 mg), DIPEA (169 μL), and 4-phenylbutyric acid (128 mg) were added at room temperature and the mixture was stirred for 3 hr. After completion of the reaction, the reaction solution was extracted with water and ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=50:50-0:100) to give the title compound (50 mg) as an orange oil.
MS (ESI) m/z: 710.3 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (226 mg) described in Reference Example 1, 4-phenylpiperidine (251 mg), and potassium carbonate (215 mg) were dissolved in NMP (1 mL), and the mixture was stirred under microwave radiation at 100° C. for 10 min, and then stirred at 160° C. for 40 min. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (186 mg) as a brown viscous substance. MS (ESI) m/z: 441.1 [M+H]+
tert-Butyl (2S,4R)-4-hydroxy-2-[4-(4-phenylpiperidine-1-carbonyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (186 mg) described in Reference Example 158 was dissolved in DMF (1.6 mL), cesium carbonate (419 mg) and methyl iodide (60 μL) were added at room temperature, and the mixture was stirred for 3 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (ethyl acetate:methanol=95:5). Then, the obtained residue was dissolved in DCM (742 μL), trifluoroacetic acid (742 μL) was added at room temperature and the mixture was stirred for 3 hr. After completion of the reaction, the reaction mixture was eluted with a cation exchange resin (PoraPak™ Rxn CX 6 cc), washed with methanol, extracted with 2 M ammonia-methanol, and concentrated under reduced pressure. The obtained residue, N-Boc-L-tert-leucine (28 mg), HATU (46 mg), and DIPEA (53 μL) were dissolved in DMF (1 mL), and the mixture was stirred at room temperature for 4 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (ethyl acetate:methanol=95:5). The obtained crude product was purified by NH thin layer silica gel chromatography to give the title compound (8.5 mg) as a colorless oil.
MS (ESI) m/z: 568.4 [M+H]+
DMF (8 mL) was added to molecular sieve 4 A (powder) (500 mg) and cesium hydroxide monohydrate (400 mg), and the mixture was stirred at room temperature for 10 min. Cyclopropylamine (0.15 mL) was added, and the mixture was stirred for 30 min. 4-Bromobutylbenzene (500 mg) was added, and the mixture was stirred at room temperature overnight and purified by silica gel column chromatography (hexane:ethyl acetate=100:0-0:100) to give the title compound (83 mg) as a pale-yellow oil.
MS (ESI) m/z: 190.1 [M+H]+
To a solution of 5-bromopentylbenzene (700 mg) in acetonitrile (6 mL) were added cyclopropylamine (0.42 mL) and potassium carbonate (425 mg) and the mixture was stirred at 70 overnight. Insoluble material was filtered off, and the filtrate was concentrated under reduced pressure and purified by NH silica gel column chromatography (hexane:ethyl acetate=100:0-0:100) to give the title compound (45 mg) as a colorless oil. MS (ESI) m/z: 204.1 [M+H]+
DMF (1 mL) was added to (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride (50 mg) described in Reference Example 122, HATU (116 mg), DIPEA (116 μL), and N-(tert-butoxycarbonyl)-L-isoleucine 0.5 hydrate (78 mg) were added, and the mixture was stirred at room temperature for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (41 mg) as a colorless oil. MS (ESI) m/z: 367.3 [M+H]+
DMF (8 mL) and cesium hydroxide monohydrate (390 mg) were added to molecular sieve 4 A (powder) (500 mg), and the mixture was stirred at room temperature for 10 min. Cyclopropylamine (0.15 mL) was added, and the mixture was stirred for 30 min. 2-Bromoethoxymethylbenzene (500 mg) was added, and the mixture was stirred at room temperature overnight. Insoluble material was collected by filtration, water was added to the filtrate and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (ethyl acetate:methanol=100:0-80:20) to give the title compound (41 mg) as a colorless oil.
MS (ESI) m/z: 192.0 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-carbamothioylpyrrolidine-1-carboxylate (200 mg) described in Reference Example 71 was dissolved in DMF (10 mL), 1-chloropropan-2-one (0.049 mL) and DIPEA (0.096 mL) were added, and the mixture was stirred at 80° C. for 5 hr. After completion of the reaction, saturated brine was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=75:25). To the obtained crudely purified product was added 4 M hydrochloric acid-dioxane (2.8 mL), and the mixture was stirred at room temperature for about 5 hr. This was dissolved in DMSO and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (10 mg) as a white powder. MS (ESI) m/z: 185.0 [M+H]+
4-[[tert-Butoxycarbonyl (ethyl)amino]methyl]benzoic acid (300 mg) was dissolved in DMF (4.0 mL), 12 M-methylamine aqueous solution (0.25 mL) and HATU (490 mg) were added at room temperature, and the mixture was stirred for 15 hr. HATU (325 mg) was added and the mixture was further stirred with heating for 25 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed successively with water and saturated brine and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=50:50-0:100). The obtained residue was dissolved in THE (2.5 mL), 4 M hydrochloric acid-dioxane (2.5 mL) was added and the mixture was stirred for 8 hr. After completion of the reaction, the reaction mixture was diluted with ethyl acetate. The precipitate was collected by filtration, washed with ethyl acetate, and dried to give the title compound (246 mg) as a white powder. MS (ESI) m/z: 193.0 [M+H]+
1,4-Dioxane (10 mL) was added to tert-butyl N-methyl-N-(4-piperidylmethyl)carbamate (500 mg), bromobenzene (0.46 mL), palladium(II) acetate (49 mg), XantPhos (127 mg), and cesium carbonate (1.4 g), and the mixture was stirred under nitrogen atmosphere at 110° C. for 5 hr. Palladium (II) acetate (49 mg) and XantPhos (127 mg) were added, and the mixture was further stirred at 110° C. for 6 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=85:15-75:25) to give the title compound (100 mg) as a pale-yellow oil. MS (ESI) m/z: 305.3 [M+H]+
tert-Butyl N-methyl-N-[(1-phenyl-4-piperidyl)methyl]carbamate (100 mg) described in Reference Example 166 was dissolved in DCM (3 mL), trifluoroacetic acid (1.5 mL) was added and the mixture was stirred for 2.5 hr. The reaction mixture was concentrated under reduced pressure, and the obtained residue was dissolved in THE (4.0 mL). 4 M Hydrochloric acid-ethyl acetate (0.50 mL) was added, and the mixture was concentrated under reduced pressure to give the title compound (110 mg) as a pale-green powder.
MS (ESI) m/z: 205.2 [M+H]+
tert-Butyl N-methyl-N-(4-piperidylmethyl)carbamate (400 mg), 2-fluorobenzaldehyde (0.46 mL) was dissolved in dichloromethane (5.0 mL), sodium triacetoxyborohydride (695 mg) was added, and the mixture was stirred for 23 hr. After completion of the reaction, the reaction mixture was extracted with saturated aqueous sodium hydrogen carbonate and chloroform. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=70:30-50:50) to give the title compound (461 mg) as a colorless oil. MS (ESI) m/z: 337.4 [M+H]+
To tert-butyl N-[(4-bromophenyl)methyl]-N-methyl-carbamate (400 mg), (2-methyl-4-pyridyl)boronic acid (270 mg), PdCl2(PPh3)2 (105 mg), and potassium carbonate (365 mg) were added water (1 mL) and 1,4-dioxane (6 mL), and the mixture was stirred under microwave radiation at 120° C. for 1 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=70:30-35:65) to give the title compound (372 mg) as a pale-brown oil. MS (ESI) m/z: 313.3 [M+H]+
tert-Butyl N-methyl-N-[[4-(2-methyl-4-pyridyl)phenyl]methyl]carbamate (372 mg) described in Reference Example 169 was dissolved in DCM (6 mL), trifluoroacetic acid (2 mL) was added and the mixture was stirred for 2 hr. The reaction mixture was concentrated under reduced pressure, the obtained residue was dissolved in THE (4.0 mL), and 4 M hydrochloric acid-ethyl acetate (1.0 mL) was added. The reaction mixture was diluted with ethyl acetate and the precipitate was collected by filtration, washed with ethyl acetate, and dried to give the title compound (395 mg) as a pale-red powder. MS (ESI) m/z: 213.2 [M+H]+
To tert-butyl N-[(4-bromophenyl)methyl]-N-methyl-carbamate (300 mg), (6-methyl-3-pyridyl)boronic acid (175 mg), PdCl2(PPh3)2 (80 mg), and potassium carbonate (275 mg) were added water (1 mL) and 1,4-dioxane (5 mL), and the mixture was stirred under microwave radiation at 120° C. for 1 hr. After completion of the reaction, the reaction mixture was extracted with water and ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate=80:20-45:55) to give the title compound (243 mg) as a pale-yellow oil. MS (ESI) m/z: 313.3 [M+H]+
tert-Butyl N-methyl-N-[[4-(6-methyl-3-pyridyl)phenyl]methyl]carbamate (243 mg) described in Reference Example 171 was dissolved in DCM (4.5 mL), trifluoroacetic acid (1.5 mL) was added and the mixture was stirred for 4 hr. The reaction mixture was concentrated under reduced pressure, the obtained residue was dissolved in THF (4.0 mL), and 4 M hydrochloric acid-ethyl acetate (0.80 mL) was added. The reaction mixture was diluted with ethyl acetate and the precipitate was collected by filtration, washed with ethyl acetate, and dried to give the title compound (175 mg) as a white powder. MS (ESI) m/z: 213.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 170 using tert-butyl N-[[1-[(2-fluorophenyl)methyl]-4-piperidyl]methyl]-N-methyl-carbamate described in Reference Example 168. MS (ESI) m/z: 237.3 [M+H]+
To a solution of 1H-tetrazole-5-ethyl acetate (500 mg) in pyridine (3 mL) was added butyryl chloride (0.37 mL) and the mixture was stirred at room temperature for 1 hr and then heated under reflux for 5 hr. After cooling to room temperature, the mixture was concentrated under reduced pressure. Ethyl acetate was added to the residue, and the residue was successively washed with 1 M hydrochloric acid and water. The organic layer was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (hexane:ethyl acetate=95:5-50:50) to give the title compound (110 mg) as a yellow oil.
MS (ESI) m/z: 199.2 [M+H]+
A solution of potassium tert-butoxide (388 mg) in DMF (3 mL) was cooled to 0° C., a solution of ethyl 2-(5-propyl-1,2,4-oxadiazol-3-yl)acetate (456 mg) in DMF (2 mL) was added and the mixture was stirred for 30 min. 2-Iodopropane (0.30 mL) was added, and the mixture was stirred for 3 hr while gradually allowing to warm to room temperature. Water was added to the reaction mixture and the mixture was extracted with ethyl acetate. The organic layer was washed with water, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane:ethyl acetate=99:1-75:25) to give the title compound (406 mg) as a colorless oil. MS (ESI) m/z: 241.0 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 175 using ethyl 2-(5-propyl-1,3,4-oxadiazol-2-yl)acetate described in Reference Example 174.
MS (ESI) m/z: 241.0 [M+H]+
To a solution of ethyl 2-(4-bromopyrazol-1-yl)-3-methylbutanoate (200 mg) described in Reference Example 31 in THE (4 mL) were added 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (167 mg), tetrakis (triphenyl phosphine) palladium (0) (84 mg), tripotassium phosphate (463 mg), and water (0.052 mL). After nitrogen substitution, and the mixture was stirred under microwave radiation at 120° C. for 30 min, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=99:1-65:35) to give the title compound (16 mg) as a pale-yellow powder. MS (ESI) m/z: 278.0 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 8 using ethyl 3-methyl-2-(5-propyl-1,2,4-oxadiazol-3-yl)butanoate described in Reference Example 175. MS (ESI) m/z: 213.0 [M+H]+
The title compound was obtained by the same reaction and treatment as in Reference Example 8 using ethyl 3-methyl-2-(5-propyl-1,3,4-oxadiazol-2-yl)butanoate described in Reference Example 176. MS (ESI) m/z: 213.0 [M+H]+
N-{5-Bromo-4-fluoro-2-[rel-(3R,5S)-3,4,5-trimethylpiperazin-1-yl]phenyl}-4-(trifluoromethyl)-6-[2-(trimethylsilyl)ethoxy]pyridine-3-carboxamide (1.0 g), benzyl {2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy]ethyl}carbamate (1.0 g), bis[tert-butyl(4-dimethylaminophenyl)phosphine]dichloropalladium(II) (120 mg), tripotassium phosphate (700 mg) were added to 1,4-dioxane (30 mL) and water (3 mL), and the mixture was stirred at 100° C. for 2 hr. The mixture was allowed to cool to room temperature, diluted with ethyl acetate, and dried over sodium sulfate. Insoluble material was filtered off, and the filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (chloroform:methanol=100:0-95:5) and NH silica gel column chromatography (hexane:ethyl acetate=80:20-50:50) purified to give the title compound (1.22 g) as a colorless solid.
MS (ESI) m/z: 796.6 [M+H]+
To a solution of Reference Example compound 180-1 (1.22 g) in methanol (30 mL) was added 10% Pd/C (500 mg), and the mixture was stirred under hydrogen atmosphere at room temperature for 2 hr. After completion of the reaction, insoluble material was filtered off with celite, and the filtrate was concentrated under reduced pressure to give the title compound (1.05 g) as a gray solid.
MS (ESI) m/z: 662.5 [M+H]+
To Reference Example compound 146 (50 mg) was added 4 M hydrochloric acid-dioxane (20 mL) at room temperature and the mixture was stirred for 0.5 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the title compound as an unpurified yellow oil (50 mg). MS (ESI) m/z: 496.2 [M+H]+
To a solution of tert-butyl [2-(2-{2-[(4-methylbenzene-1-sulfonyl) oxy]ethoxy}ethoxy)ethoxy]acetate (6.0 g), N-{1,3-dimethyl-2-oxo-6-[(propan-2-yl)oxy]-2,3-dihydro-1H-benzoimidazol-5-yl}-4-hydroxy-2-methoxybenzamide (5.53 g), and N,N-dimethylformamide (100 mL) was added potassium carbonate (5.94 g) and the mixture was stirred at 60° C. for 12 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (formic acid addition) to give the title compound (3.0 g) as a yellow oil.
MS (ESI) m/z: 386.2 [M+H]+
To a solution of Reference Example compound 182-1 (3.0 g) in DCM (20 mL) was added trifluoroacetic acid (5.0 mL) and the mixture was stirred with heating at room temperature for 19 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (water (0.1% TFA)-acetonitrile) to give the title compound (1.0 g) as a colorless oil.
MS (ESI) m/z: 576.3 [M+H]+
Using Reference Example compound 18, the same reaction and treatment as in Reference Example 70 were performed, and then the same reaction and treatment as in Reference Example 96 were performed using 5-benzyl-5-oxopentanoic acid to give the title compound (25 mg) as a pale-yellow solid.
MS (ESI) m/z: 644.3 [M+H]+
2-[(2S,4R)-4-Hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8, HATU (30 mg), and 4-phenoxypiperidine (14 mg) were dissolved in DMF (600 μL), DIPEA (40 μL) was added and the mixture was stirred at room temperature for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (16 mg) as a white powder. MS (ESI) m/z: 536.3 [M−H]−
2-[(2S,4R)-4-Hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8, HATU (30 mg), and 4-phenylpiperidine (13 mg) were dissolved in DMF (600 μL), DIPEA (36 μL) was added and the mixture was stirred at room temperature for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (20 mg) as a white powder.
MS (ESI) m/z: 520.3 [M−H]−
HATU (30 mg) and 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8 was dissolved in DMF (500 μL), were stirred for about 10 min. N-(cyclobutylmethyl)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (18 mg) described in Reference Example 10 and DIPEA (27 μL) were added and the mixture was stirred at room temperature for about 2 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (27 mg) as a white powder.
MS (ESI) m/z: 633.3 [M+H]+
HATU (30 mg) and 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8 were dissolved in DMF (500 μL), and the mixture was stirred for about 10 min. 2-Methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]propan-1-amine (17 mg) described in Reference Example 11 and DIPEA (27 μL) were added and the mixture was stirred at room temperature for about 2 hr. To the reaction mixture were added chloroform and saturated aqueous sodium hydrogen carbonate, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile). The obtained crude product was further purified by silica gel column chromatography (ethyl acetate:methanol=100:0-85:15) to give the title compound (15 mg) as a white powder. MS (ESI) m/z: 621.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 3 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(4-phenylphenyl)methanamine. MS (ESI) m/z: 532.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 3 using (3R,5S)-5-[5-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]-1H-imidazol-2-yl]pyrrolidin-3-ol hydrochloride described in Reference Example 18 and (2S)-2-acetamido-3,3-dimethylbutanoic acid. MS (ESI) m/z: 482.3 [M+H]+
2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (9.5 mg) described in Reference Example 14 was dissolved in NMP (1.0 mL), HATU (30 mg) was added and the mixture was stirred. DIPEA (14 μL) and 2-(2-pyridyl)ethaneamine (5.0 mg) were added and the mixture was stirred at room temperature for about 30 min. The reaction mixture was adsorbed onto PoraPak™ Rxn Rp and washed with water. After elution with acetonitrile-water, the eluate was concentrated by centrifugation. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=95:5-70:30) to give the title compound (4.6 mg) as a colorless powder. MS (ESI) m/z: 457.3 [M+H]+
2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (13 mg) described in Reference Example 14 was dissolved in NMP (4.0 mL), HATU (30 mg) was added and the mixture was stirred for about 10 min. N-methyl-4-phenyl-butan-1-amine (7.1 mg) and DIPEA (18 L) were added and the mixture was stirred at room temperature for about 10 min. The reaction mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (9.4 mg) as a colorless oil. MS (ESI) m/z: 498.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 7 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 2-phenylethaneamine. MS (ESI) m/z: 456.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 7 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 2-(2-methoxyphenyl)ethaneamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 7 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 2-(3-methoxyphenyl)ethaneamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 7 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 2-(4-methoxyphenyl)ethaneamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1,2,3,4-tetrahydro-2,7-naphthyridine dihydrochloride.
MS (ESI) m/z: 469.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(2-chlorophenyl)-N-methyl-methanamine.
MS (ESI) m/z: 490.2/492.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(3-chlorophenyl)-N-methyl-methanamine.
MS (ESI) m/z: 490.2/492.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(4-chlorophenyl)-N-methyl-methanamine.
MS (ESI) m/z: 490.2/492.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(2-fluorophenyl)-N-methyl-methanamine. MS (ESI) m/z: 474.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(3-fluorophenyl)-N-methyl-methanamine. MS (ESI) m/z: 474.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(4-fluorophenyl)-N-methyl-methanamine. MS (ESI) m/z: 474.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-[2-(trifluoromethyl)phenyl]methanamine.
MS (ESI) m/z: 524.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-[3-(trifluoromethyl)phenyl]methanamine.
MS (ESI) m/z: 524.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-[4-(trifluoromethyl)phenyl]methanamine.
MS (ESI) m/z: 524.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(2-methoxyphenyl)-N-methyl-methanamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(3-methoxyphenyl)-N-methyl-methanamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 1-(4-methoxyphenyl)-N-methyl-methanamine. MS (ESI) m/z: 486.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-pyrimidin-2-yl-methanamine. MS (ESI) m/z: 458.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(1-methylpyrazol-3-yl)methanamine. MS (ESI) m/z: 460.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-3-phenyl-propan-1-amine. MS (ESI) m/z: 484.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-[3-(4-methylthiazol-5-yl)phenyl]methanamine described in Reference Example 20. MS (ESI) m/z: 553.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(2-naphthyl)methanamine. MS (ESI) m/z: 506.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 8 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(1-naphthyl)methanamine. MS (ESI) m/z: 506.3 [M+H]+
4 M Hydrochloric acid-dioxane (2.0 mL) was added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-phenylphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (34 mg) described in Reference Example 22, and the mixture was stirred at room temperature for about 14 hr. After completion of the reaction, the precipitated solid was collected by filtration. To the collected powder were added dichloromethane (1.0 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (16 mg), HATU (30 mg), and DIPEA (30 μL), and the mixture was stirred at room temperature for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (22 mg) as a white powder. MS (ESI) m/z: 501.6 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[3-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (35 mg) described in Reference Example 24 was dissolved in 1,4-dioxane (1.0 mL), 4 M hydrochloric acid-dioxane (200 μL) was added, and the mixture was stirred at room temperature for about 16 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the residue were added dichloromethane (1.0 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (16 mg), HATU (29 mg), and DIPEA (40 μL), and the mixture was stirred at room temperature for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (26 mg) as a white powder. MS (ESI) m/z: 522.3 [M+H]+
4 M Hydrochloric acid-dioxane (1.0 mL) was added to tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[2-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (70 mg) described in Reference Example 26, and the mixture was stirred at room temperature for about 2 hr. After completion of the reaction, the precipitated solid was collected by filtration. To the collected powder were added dichloromethane (1.0 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (32 mg), HATU (55 mg), and DIPEA (60 μL), and the mixture was stirred at room temperature for about 1.5 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (5.6 mg) as a white powder. MS (ESI) m/z: 536.3 [M+H]+
2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 (23 mg), N-methyl-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (22 mg), DCM (1252 μL), DIPEA (16 μL), and HATU (30 mg) were successively added and the mixture was stirred at room temperature overnight. N-Methyl-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (22 mg) and DIPEA (16 μL) were added and the mixture was stirred at room temperature for about 2 hr. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-75:25). Furthermore, the obtained crude product was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (11 mg) as a white powder. MS (ESI) m/z: 553.3 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (17 mg) described in Reference Example 8, N-(cyclopropylmethyl)-4-phenyl-butan-1-amine (27 mg) described in Reference Example 27, and HATU (22 mg) were dissolved in DCM (1 mL), DIPEA (30 μL) was added, and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (10 mg) as a white powder. MS (ESI) m/z: 564.6 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (18 mg) described in Reference Example 8, N-(cyclopropylmethyl)-4-(4-pyridyl)butan-1-amine (12 mg) described in Reference Example 28, and HATU (25 mg) was dissolved in DCM (1 mL), DIPEA (30 μL) was added, and the mixture was stirred at room temperature for 2.5 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (15 mg) as a white powder.
MS (ESI) m/z: 565.7 [M+H]+
N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]propane-2-amine (27 mg) described in Reference Example 29, 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8, and HATU (26 mg) was dissolved in DCM (1 mL), DIPEA (30 μL) was added, and the mixture was stirred at room temperature for 2.5 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (14 mg) as a white powder.
MS (ESI) m/z: 607.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 38 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 4,5,6,7-tetrahydrothiazolo-[5,4-c]pyridine salt acid.
MS (ESI) m/z: 475.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 38 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 2,2,2-trifluoro-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]ethaneamine described in Reference Example 30.
MS (ESI) m/z: 647.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 38 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-methyl-1-(4-pyridyl)piperidine-4-amine.
MS (ESI) m/z: 552.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 38 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-methyl-1-(4-phenylthiazol-2-yl)methanamine.
MS (ESI) m/z: 565.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 38 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-methyl-1-[1-(4-pyridyl)-4-piperidyl]methanamine.
MS (ESI) m/z: 566.6 [M+H]+
(3R,5S)-5-[5-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]-1H-imidazol-2-yl]pyrrolidin-3-ol hydrochloride (30 mg) described in Reference Example 18 was dissolved in DMF (3 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (18 mg), DIPEA (57 μL), and HATU (41 mg) were added, and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, the reaction was quenched by adding saturated brine, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (29 mg) as a colorless powder. MS (ESI) m/z: 508.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 44 using (3R,5S)-5-[5-[3-(4-methyl-1,3-thiazol-5-yl)phenyl]-1H-imidazol-2-yl]pyrrolidin-3-ol hydrochloride described in Reference Example 18 and 2-[4-(2-methoxypyridin-4-yl)pyrazol-1-yl]-3-methylbutanoic acid described in Reference Example 33. MS (ESI) m/z: 584.4 [M+H]+
A solution (0.10 mL) of 2-pyridylmethanamine (121 mg) and DIPEA (0.60 mL) in NMP (3.4 mL), and a solution (0.65 mL) of 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 (120 mg) and HATU (142 mg) in NMP (8 mL) were injected into a flow reactor, mixed and reacted at 100° C. for 10 min. The reaction mixture was adsorbed onto PoraPak™ Rxn Rp and washed with water. The eluate was concentrated by centrifugation and the residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (2.7 mg) as a pale-yellow oil. MS (ESI) m/z: 443.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and 3-pyridylmethanamine. MS (ESI) m/z: 443.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(2-pyridyl)methanamine. MS (ESI) m/z: 457.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(4-pyridyl)methanamine. MS (ESI) m/z: 457.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and phenylmethanamine. MS (ESI) m/z: 442.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-phenyl-methanamine. MS (ESI) m/z: 456.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-2-phenylethaneamine. MS (ESI) m/z: 470.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 46 using 2-[(2S,4R)-1-[(2S)-2-acetamido-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 14 and N-methyl-1-(3-pyridyl)methanamine. MS (ESI) m/z: 457.3 [M+H]+
2-[(2S,4R)-4-hydroxypyrrolidin-2-yl]-N-methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide hydrochloride (40 mg) described in Reference Example 6, 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (24 mg), and HOBt (17 mg) were dissolved in DCM (420 μL), Et3N (35 μL) and WSC·HCl (24 mg) were added, and the mixture was stirred at room temperature for 2.5 hr. MeOH was added and the mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (18 mg) as a white powder. MS (ESI) m/z: 577.3 [M−H]−
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide (10 mg) described in Reference Example 34 was separated using a chiral column [CHIRALPAK IC(30*250), methanol]. The fraction eluted earlier was recovered, and concentrated under reduced pressure to give the title compound (3.5 mg) as a white powder.
MS (ESI) m/z: 579.3 [M+H]+
HPLC conditions described in the experiment term of 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-methyl-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide (10 mg) described in Reference Example 34 was separated using a chiral column [CHIRALPAK IC(30*250), methanol]. The fraction eluted later was recovered, and concentrated under reduced pressure to give the title compound (2.8 mg) as a white powder.
MS (ESI) m/z: 579.3 [M+H]+
chiral HPLC conditions described in the experiment term of N-(cyclopropylmethyl)-2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide
N-(cyclopropylmethyl)-2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1H-imidazole-4-carboxamide (14 mg) described in Reference Example 36 was separated using a chiral column [CHIRALPAK IE(30*250), methyl tert-butyl ether:methanol=87:13]. The fraction eluted earlier was recovered, and concentrated to give the title compound (4 mg) as a white solid. MS (ESI) m/z (ESI) m/z: 619.3 [M+H]+
N-(cyclopropylmethyl)-2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl] methyl]-1H-imidazole-4-carboxamide (14 mg) described in Reference Example 36 was separated using a chiral column [CHIRALPAK IE(30*250), methyl tert-butyl ether:methanol=87:13]. The fraction eluted later was recovered, and concentrated to give the title compound (4 mg) as a white solid. MS (ESI) m/z (ESI) m/z: 619.3 [M+H]+
tert-Butyl (2S,4R)-2-[4-[cyclopropylmethyl-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate (26 mg) described in Reference Example 35 was dissolved in 1,4-dioxane (0.4 mL), 4 M hydrochloric acid-dioxane (0.4 mL) was added at room temperature, and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled 3 times with toluene. The obtained residue was dissolved in N,N-dimethylformamide (0.5 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (13 mg), HATU (22 mg), and diisopropyl ethylamine (34 μL) were added, and the mixture was stirred at room temperature for 3 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate, and concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (chloroform:methanol=100:0-90:10) to give the title compound (19 mg) as a white powder. MS (ESI) m/z: 617.4 [M−H]−
tert-Butyl N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-N-[2-(oxan-2-yloxy)ethyl]carbamate (18 mg) described in Reference Example 37 was dissolved in 1,4-dioxane (0.4 mL), 4 M hydrochloric acid-dioxane (0.2 mL) was added at room temperature, and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and azeotropically distilled 3 times with toluene. The obtained residue was dissolved in N,N-dimethylformamide (0.5 mL), 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (16 mg), HATU (24 mg), and diisopropyl ethylamine (30 μL) were added, and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (14 mg) as a colorless oil. MS (ESI) m/z: 607.4 [M−H]−
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-(3-methoxyphenyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (27 mg) described in Reference Example 40 was dissolved in 1,4-dioxane (1 mL), 4 M hydrochloric acid-dioxane (0.5 mL) was added at room temperature and the mixture was stirred for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, 1,4-dioxane (10 mL) was added and the mixture was concentrated. The obtained residue was dissolved in N,N-dimethylformamide (1.1 mL), diisopropyl ethylamine (39 μL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (15 mg), and HATU (28 mg) were added, and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, water and saturated brine were added, and the mixture was extracted with ethyl acetate and hexane and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=95:5-70:30), and again purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (7 mg) as a white powder. MS (ESI) m/z: 441.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-hydroxy-2-[5-[(4-methoxyphenyl)methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 41 and 2-(3-methoxy-1,2-oxazol-5-yl)-3 methylbutanoic acid. MS (ESI) m/z: 455.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid and tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[(3-methoxyphenyl)methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 42. MS (ESI) m/z: 455.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[2-(methylcarbamoyl)phenyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 4.3 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 468.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using (3R,5S)-5-[1-[(3-phenoxyphenyl)methyl]imidazol-2-yl]pyrrolidin-3-ol hydrochloride described in Reference Example 45 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid.
MS (ESI) m/z: 517.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(2-phenylmethoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 46 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 531.6 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-phenylmethoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 47 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 531.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[1-[(3-phenylmethoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 47 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 531.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid described in Reference Example 48 and tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(2-phenoxyphenyl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate.
MS (ESI) m/z: 517.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-2-[4-[ethyl-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 49 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 593.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-hydroxy-2-[4-[methyl-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl] ethyl] carbamoyl]-1H-imidazol-2-yl] pyrrolidine-1-carboxylate described in Reference Example 52 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 593.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[2-[(4-methyl-1,3-thiazol-5-yl)methoxy]phenyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 54 and 2-(3-methoxy-1,2-oxazol-5-yl)-3 methylbutanoic acid. MS (ESI) m/z: 552.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[3-[(4-methyl-1,3-thiazol-5-yl)methoxy]phenyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 56 and 2-(3-methoxy-1,2-oxazol-5-yl)-3 methylbutanoic acid. MS (EST) m/z: 552.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid and tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-(4-methoxyphenyl)-4-(trifluoromethyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 58. MS (ESI) m/z: 509.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-hydroxy-2-[5-(4-methoxyphenyl)-4-(methylcarbamoyl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 60 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid.
MS (ESI) m/z: 498.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid and tert-butyl (2S,4R)-2-[4,5-bis(3-methoxyphenyl)-1H-imidazol-2-yl]-4-[tert-butyl(dimethyl)silyl]oxypyrrolidine-1-carboxylate described in Reference Example 61. MS (ESI) m/z: 547.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-(naphthalen-1-ylmethyl)imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 62 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid.
MS (ESI) m/z: 475.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid and tert-butyl-[(3R,5S)-5-[5-(4-methoxyphenyl)-1H-imidazol-2-yl]pyrrolidin-3-yl]oxy-dimethylsilane described in Reference Example 64.
MS (ESI) m/z: 441.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid and tert-butyl (2S,4R)-4-hydroxy-2-[5-[[4-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 67.
MS (ESI) m/z: 493.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[5-[3-(methylcarbamoyl)phenyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 68 and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 468.2 [M+H]+
tert-Butyl N-[(2S)-1-[(2S,4R)-2-[5-[(4-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (34 mg) described in Reference Example 70 was dissolved in 1,4-dioxane (3 mL), 4 M hydrochloric acid-dioxane (0.5 mL) was added at room temperature, and the mixture was stirred for 6 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and then azeotropically distilled with toluene. The obtained residue was dissolved in dichloromethane (5 mL), triethylamine (53 μL) and acetic anhydride (12 μL) were added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, 1 M sodium hydroxide aqueous solution was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=90:10) to give the title compound (16 mg) as a colorless powder.
MS (ESI) m/z: 477.2/479.2 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[2-(4-chlorophenyl)ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (52 mg) described in Reference Example 73 was dissolved in dichloromethane (2 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and then azeotropically distilled twice with toluene. The obtained residue was dissolved in DMF (3 mL), DIPEA (89 μL), (2S)-2-acetamido-3,3-dimethylbutanoic acid (29 mg), and HATU (51 mg) were added, and the mixture was stirred at room temperature for 30 min. The reaction mixture was diluted with ethyl acetate, and the organic layer was washed with water and saturated brine and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in 1,4-dioxane (2 mL), 4 M hydrochloric acid-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hr. The reaction mixture was concentrated under reduced pressure, and the obtained residue was suspended in THE (3 mL). Triethylamine (71 μL) and acetic anhydride (29 μL) were added, and the mixture was stirred at room temperature for 30 min. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (26 mg) as a pale-yellow oil.
MS (ESI) m/z: 447.4/449.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-2-[5-[(2-chlorophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 76.
MS (ESI) m/z: 433.4/435.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[5-[2-(trifluoromethyl)phenyl]-1H-imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 78. MS (ESI) m/z: 453.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[5-[[2-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 83. MS (ESI) m/z: 465.2 [M−H]−
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[5-[[3-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 84. MS (ESI) m/z: 467.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[1-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 87.
MS (ESI) m/z: 496.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-2-[5-(4-bromophenyl)-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 88. MS (ESI) m/z: 463.3/465.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[5-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]-1H-imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 89.
MS (ESI) m/z: 482.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-2-[5-[(3-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 92.
MS (ESI) m/z: 477.3/479.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(1S)-1-[(2S,4R)-4-hydroxy-2-[5-[[3-(4-methylthiazol-5-yl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate described in Reference Example 94.
MS (ESI) m/z: 496.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[5-[[4-(trifluoromethyl)phenyl]methyl]-1H-imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate described in Reference Example 95. MS (ESI) m/z: 467.3 [M+H]+
N-[(2S)-1-[(2S,4R)-2-[5-[(4-bromophenyl)methyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]acetamide (13 mg) described in Reference Example 96 was dissolved in 1,4-dioxane (2 mL) and water (0.5 mL), and 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (9.2 mg), and sodium carbonate (8 mg) were added at room temperature. After deaeration, a 1,1′-(diphenylphosphino)ferrocene-palladium(II) dichloromethane complex (2 mg) was added, and the mixture was stirred under nitrogen atmosphere at 80° C. for 1 hr. After completion of the reaction, anhydrous sodium sulfate was added. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (5 mg) as a pale-brown powder.
MS (ESI) m/z: 496.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 94 using N-[(2S)-1-[(2S,4R)-2-[1-[2-(4-chlorophenyl)ethyl]imidazol-2-yl]-4-hydroxypyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]acetamide described in Reference Example 97 and 4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole. MS (ESI) m/z: 510.3 [M+H]+
Methyl 2-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1-[[3-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]imidazole-4-carboxylate (83 mg) described in Reference Example 99 was dissolved in methanol (1 mL), THE (1 mL) and 1 M sodium hydroxide aqueous solution (1 mL) were added and the mixture was stirred at 50° C. for 17 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. THE and 1 M hydrochloric acid (1 mL) were added, and the mixture was concentrated again under reduced pressure. The obtained residue was dissolved in dichloromethane (1 mL), diisopropyl ethylamine (70 μL), and HATU (67 mg), N-methyl-N-(4-pyridylmethyl)amine (20 μL) were added, and the mixture was stirred at room temperature for 1.5 hr. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was dissolved in 1,4-dioxane (0.7 mL), 4 M hydrochloric acid-dioxane (0.3 mL) was added at room temperature, and the mixture was stirred for 2 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was dissolved in dichloromethane (1 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (33 mg), HATU (67 mg), and DIPEA (70 μL) were added, and the mixture was stirred at room temperature for 17 hr. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (20 mg) as a pale-yellow powder. MS (ESI) m/z: 670.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 95 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-(2-methoxy-2-oxoethyl)imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 100, 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid, and aniline instead of N-methyl-N-(4-pyridylmethyl)amine.
MS (ESI) m/z: 468.3 [M+H]+
4-[2-[(2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidin-2-yl]-1H-imidazol-5-yl]benzoic acid (25 mg) described in Reference Example 101 was dissolved in N,N-dimethylformamide (2 mL), triethylamine (21 μL), methylamine hydrochloride (4 mg), and HATU (23 mg) were added and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was dissolved in 1,4-dioxane (1 mL), 4 M hydrochloric acid-dioxane (1 mL) was added at room temperature, and the mixture was stirred for 1 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was dissolved in N,N-dimethylformamide (2 mL), triethylamine (0.2 mL), 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (11 mg), and HATU (23 mg) were added and the mixture was stirred at room temperature for 18 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (0.40 mg) as a pale-yellow oil.
MS (ESI) m/z: 468.2 [M+H]+
4 M Hydrochloric acid-dioxane (1 mL) was added to tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[1-[(4-methoxyphenyl)methyl]-4-(phenylmethoxymethyl)imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (14 mg) described in Reference Example 105 at room temperature, and the mixture was stirred for 1 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was dissolved in dichloromethane (0.5 mL), acetic anhydride (3 μL) and triethylamine (15 μL) were added and the mixture was stirred at room temperature for 2 hr. After completion of the reaction, potassium carbonate (10 mg) and methanol (200 μL) were added, and the mixture was stirred at room temperature for 30 min. Water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was dissolved in acetonitrile (1 mL), water (0.3 mL) and ammonium hexanitratocerate(IV) (92 mg) were added, and the mixture was stirred at room temperature for 3.5 hr. After completion of the reaction, water and chloroform were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (1.1 mg) as a colorless oil. MS (ESI) m/z: 429.3 [M+H]+
3-Phenoxypropan-1-amine (2 g), acetone (20 mL), and molecular sieve (5 g) were combined and the mixture was stirred at room temperature for 2 days. The reaction solution was filtered, washed with methanol, and the filtrate was concentrated under reduced pressure. The concentrated residue was dissolved in methanol (30 mL), sodium borohydride (250 mg) was added at room temperature, and the mixture was stirred as it was. The reaction solution was concentrated under reduced pressure, chloroform and water were added, and the mixture was extracted with chloroform. The organic layer was washed with water and saturated brine, dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. To the concentrated residue was added 30% hydrochloric acid-ethanol (1 mL), and the mixture was filtered and recrystallized from ethanol. 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (25 mg) described in Reference Example 8 was dissolved in DMF (1 mL), HATU (38 mg), the earlier precipitate (29 mg), and DIPEA (40 μL) were added at room temperature, and the mixture was stirred at room temperature for 16 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (17 mg) as a white powder.
MS (ESI) m/z: 554.3 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(1R)-1-(1-naphthyl)ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (152 mg) described in Reference Example 118 was dissolved in methanol (1460 μL), 4 M hydrochloric acid-dioxane (730 μL) was added at room temperature and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, purified by preparative HPLC (0.05% trifluoroacetic acid aqueous solution-0.05% trifluoroacetic acid acetonitrile solution), and the solvent was evaporated. To the obtained residue were added, at room temperature, Et3N (74 μL), and a solution of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (46 mg), HOBt (36 mg), and WSC·HCl (51 mg) in DMF (1.8 mL), and the mixture was stirred for 6 hr. After completion of the reaction, water (100 μL) was added and the mixture was diluted with DMSO, and purified by preparative HPLC (0.05% trifluoroacetic acid aqueous solution-0.05% trifluoroacetic acid acetonitrile solution) to give the title compound (69 mg) as a white powder. MS (ESI) m/z: 489.5 [M+H]+
tert-Butyl (2S,4R)-4-hydroxy-2-[4-methyl-5-[methyl-[[4-(4-methylthiazol-5-yl)phenyl]methyl]carbamoyl]-1H-imidazol-2-yl]pyrrolidine-1-carboxylate (14 mg) described in Reference Example 117 was dissolved in methanol (547 μL), 4 M hydrochloric acid-dioxane (68 μL) was added at room temperature and the mixture was stirred for 1 day. Furthermore, 4 M hydrochloric acid-dioxane (68 μL) was added and the mixture was stirred for 1 day. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and azeotropically distilled with toluene. Separately, to a solution of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (7.0 mg) and HOBt (5.5 mg) in DMF (0.27 mL) was added WSC·HCl (7.8 mg) at room temperature, and the mixture was stirred for 20 min. This solution was added to a mixture of the earlier concentrated residue and triethylamine (11 μL) at room temperature, and the mixture was stirred for 6 hr. After completion of the reaction, water (100 μL) was added and the mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (13 mg) as a white powder.
MS (ESI) m/z: 593.5 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[2-[4-(trifluoromethyl)phenyl]-4-pyridyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (80 mg) described in Reference Example 119 was dissolved in methanol (660 μL), 4 M hydrochloric acid-dioxane (165 μL) was added at room temperature and the mixture was stirred for 2 days. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene. Separately, to a solution of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (34 mg) and HOBt (27 mg) in DMF (1.3 mL) was added WSC·HCl (38 mg) at room temperature, and the mixture was stirred for 20 min. This solution was added to a mixture of the earlier concentrated residue and triethylamine (55 μL) at room temperature, and the mixture was stirred for 6 hr. After completion of the reaction water (100 μL) was added and the mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (53 mg) as a white powder. MS (ESI) m/z: 570.5 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (25 mg) described in Reference Example 8 was dissolved in DMF (0.66 mL), HATU (38 mg) was added at room temperature and the mixture was stirred for 30 min. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]cyclopropaneamine (20 mg) described in Reference Example 134 and DIPEA (0.045 mL) were added, and the mixture was stirred at room temperature for 2 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (30 mg) as a white powder.
MS (ESI) m/z: 605.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid so (25 mg) described in Reference Example 8 was dissolved in DMF (0.66 mL), HATU (38 mg) was added at room temperature and the mixture was stirred for 30 min. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]tetrahydrofuran-3-amine (22 mg) described in Reference Example 135 and DIPEA (46 μL) were added, and the mixture was stirred at room temperature for 3 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (24 mg) as a white powder.
MS (ESI) m/z: 635.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (13 mg) described in Reference Example 8 was dissolved in DMF (0.34 mL), HATU (20 mg) was added at room temperature and the mixture was stirred for 30 min. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]tetrahydropyran-4-amine (10 mg) described in Reference Example 136 and DIPEA (24 μL) were added, and the mixture was stirred at room temperature for about 5 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (15 mg) as a white powder.
MS (ESI) m/z: 649.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (25 mg) described in Reference Example 8 was dissolved in DMF (0.66 mL), HATU (38 mg) was added at room temperature and the mixture was stirred for 30 min. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]cyclobutanamine (18 mg) described in Reference Example 137 and DIPEA (46 LL) were added, and the mixture was stirred at room temperature for 2 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) and then the obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-85:15) to give the title compound (24 mg) as a white powder.
MS (ESI) m/z: 619.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (20 mg), 3-phenoxypyrrolidine hydrochloride (11 mg), HATU (30 mg), DIPEA (36 μL), and DMF (0.60 mL) were mixed at room temperature and stirred for 4 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (17 mg) as a white powder.
MS (ESI) m/z: 524.3 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8 was dissolved in DMF (0.6 mL), HATU (30 mg), 3-phenylazetidine (7.7 mg), and DIPEA (36 μL) were added at room temperature, and the mixture was stirred for about 4 hr. After completion of the reaction, chloroform and water were added, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (3.8 mg) as a white powder. MS (ESI) m/z: 494.3 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (25 mg) described in Reference Example 8 was dissolved in DMF (0.70 mL), HATU (38 mg) was added at room temperature and the mixture was stirred for 30 min. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]cyclohexanamine (19 mg) described in Reference Example 139 and DIPEA (0.045 mL) were added, and the mixture was stirred at room temperature for 2 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) and the obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-85:15) to give the title compound (12 mg) as a white powder.
MS (ESI) m/z: 647.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (25 mg) described in Reference Example 8 was dissolved in DMF (0.70 mL), HATU (38 mg) was added at room temperature and the mixture was stirred for 1 hr. Then, N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]cyclopentanamine (19 mg) described in Reference Example 138 and DIPEA (46 μL) were added, and the mixture was stirred at room temperature for 2 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile), and the crude product was purified by preparative HPLC (0.05% trifluoroacetic acid aqueous solution-0.05% trifluoroacetic acid acetonitrile solution) to give the title compound (26 mg) as a white powder. MS (ESI) m/z: 633.5 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8 was dissolved in DMF (0.60 mL), HATU (30 mg), 3-phenylpyrrolidine (8.6 mg), and DIPEA (36 μL) were added at room temperature, and the mixture was stirred at room temperature for about 4 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (11 mg) as a white powder.
MS (ESI) m/z: 508.2 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (51 mg) described in Reference Example 21 was dissolved in 1,4-dioxane (800 μL), 4 M hydrochloric acid-dioxane (200 μL) was added at room temperature and the mixture was stirred for 20 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (43 mg), DIPEA (100 μL), HATU (68 mg), and DCM (1 mL) at room temperature and the mixture was stirred for 2.5 hr. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (23 mg) as a white powder. MS (ESI) m/z: 335.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 33 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[[5-(4-fluorophenyl)-3-pyridyl]methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 121. MS (ESI) m/z: 520.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(1S)-1-[(2S,4R)-2-[4-[cyclopropylmethyl(4-phenylbutyl)carbamoyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate described in Reference Example 146.
MS (ESI) m/z: 538.6 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[1-[2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]imidazol-2-yl]pyrrolidine-1-carboxylate (21 mg) described in Reference Example 124 was dissolved in 1,4-dioxane (800 μL), 4 M hydrochloric acid-dioxane (200 μL) was added at room temperature and the mixture was stirred for 17 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added, at room temperature, 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (19 mg), DIPEA (50 μL), HATU (32 mg), and DMF (1 mL), and the mixture was stirred for 2.5 hr. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (16 mg) as a white powder. MS (ESI) m/z: 536.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 61 using tert-butyl (2S,4R)-2-(1H-benzimidazol-2-yl)-4-hydroxypyrrolidine-1-carboxylate described in Reference Example 123, and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 385.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate described in Reference Example 125, and 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid. MS (ESI) m/z: 493.1 [M+H]+
tert-Butyl (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[1-[(3,5-dimethyl-1-phenylpyrazol-4-yl)methyl]imidazol-2-yl]pyrrolidine-1-carboxylate (25 mg) described in Reference Example 126 was dissolved in 1,4-dioxane (800 μL), 4 M hydrochloric acid-dioxane (200 μL) was added at room temperature and the mixture was stirred for 20 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added, at room temperature, 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (20 mg), DIPEA (50 μL), HATU (25 mg), and DMF (1 mL), and the mixture was stirred for 2.5 hr. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (12 mg) as a white powder. MS (ESI) m/z: 519.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 2 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8, and dimethylamine. MS (ESI) m/z: 406.4 [M+H]+
To (2S)-2-amino-1-[(2S,4R)-4-hydroxy-2-[1-[[3-(4-methylthiazol-5-yl)phenyl]methyl]imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethylbutan-1-one hydrochloride (20 mg) described in Reference Example 128 were added DCM (1 mL), Et3N (21 μL), and acetic anhydride (4.3 μL) at room temperature and the mixture was stirred for 30 min. The reaction was discontinued by adding saturated aqueous sodium hydrogen carbonate and the reaction mixture was extracted with chloroform. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (11 mg) as a white powder. MS (ESI) m/z: 496.1 [M+H]+
DCM (1 mL), Et3N (21 μL), and acetic anhydride (4.5 μL) were added to (2S)-2-amino-1-[(2S,4R)-4-hydroxy-2-[1-[2-[3-(4-methylthiazol-5-yl)phenyl]ethyl]imidazol-2-yl]pyrrolidin-1-yl]-3,3-dimethylbutan-1-one hydrochloride (20 mg) described in Reference Example 130 at room temperature and the mixture was zo stirred for 30 min. The reaction was discontinued by adding saturated aqueous sodium hydrogen carbonate, and the reaction mixture was extracted with chloroform. The organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (11 mg) as a white powder. MS (ESI) m/z: 510.2 [M+H]+
N-(2-aminoethyl)-2-[(2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-N-(4-phenylbutyl)-1H-imidazole-4-carboxamide hydrochloride (20 mg) described in Reference Example 143 was dissolved in DCM (0.60 mL), Et3N (16 μL) and acetic anhydride (3 μL) were added at room temperature and the mixture was stirred for 2 hr. The reaction was quenched by adding sodium bicarbonate water, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (4.5 mg) as a colorless viscous substance. MS (ESI) m/z: 613.3 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 60 using tert-butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl (dimethyl)silyl]oxy-2-[4-[[cyclopropylmethyl (4-phenylbutyl)amino]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-1-propyl]carbamate described in Reference Example 145, 1-fluorocyclopropanecarboxylic acid, and methanol instead of 1,4-dioxane. MS (ESI) m/z: 568.4 [M+H]+
2-[(2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (30 mg) described in Reference Example 133, and tert-butyl N-methyl-N-[2-(4-phenylbutylamino)ethyl]carbamate (28 mg) described in Reference Example 144 were dissolved in DMF (0.80 mL), DIPEA (52 μL) and HATU (37 mg) were added at room temperature and the mixture was stirred for 2 hr. Then, diisopropyl ethylamine (26 μL), HATU (20 mg), and tert-butyl N-methyl-N-[2-(4-phenylbutylamino)ethyl]carbamate (14 mg) were added and the mixture was stirred for 20 hr. The reaction mixture was diluted with DMSO and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile). The residue was dissolved in 1,4-dioxane (0.88 mL), 4 M hydrochloric acid-dioxane (0.11 mL) was added at room temperature and the mixture was stirred for 2 hr. 4 M Hydrochloric acid-dioxane (0.22 mL) was added and the mixture was stirred for 20 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled 3 times with toluene. The obtained residue was dissolved in dichloromethane (0.7 mL), acetic anhydride (8 μL) and triethylamine (19 μL) were added and the mixture was stirred at room temperature for 1 hr. To the reaction mixture was added sodium bicarbonate water and the mixture was extracted with chloroform. The organic layer was separated and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (10 mg) as a white powder.
MS (ESI) m/z: 627.4 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-2-[5-[2-(3-chlorophenyl)ethyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carbonyl]-2,2-dimethylpropyl]carbamate (74 mg) described in Reference Example 113 was dissolved in 1,4-dioxane (3 mL), 4 M hydrochloric acid-dioxane (1 mL) was added at room temperature and the mixture was stirred for 6 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene. To the obtained residue were added, at room temperature, DCM (4 mL), Et3N (82 μL), and acetic anhydride (20.7 μL) and the mixture was stirred for 2 hr. After completion of the reaction, the reaction was discontinued by adding 1 M sodium hydroxide aqueous solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The organic layer was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (56 mg) as a pale-yellow powder.
MS (ESI) m/z: 447.2/449.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 81 using tert-butyl N-[(1S)-1-[(2S,4R)-2-[5-[2-(4-chlorophenyl)ethyl]-1H-imidazol-2-yl]-4-hydroxypyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate described in Reference Example 109.
MS (ESI) m/z: 447.4/449.4 [M+H]+
2-(3-Methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (6.3 mg), HOBt (4.7 mg), and WSC·HCl (6.8 mg) were dissolved in DMF (77 μL), and the mixture was stirred at room temperature for 30 min. The reaction mixture was added to a solution of (3R,5S)-5-[1-(2-naphthylmethyl)imidazol-2-yl]pyrrolidin-3-ol (8.4 mg) described in Reference Example 151 and Et3N (12 μL) in DMF (200 μL), and the mixture was stirred at room temperature for 5 hr. The reaction was discontinued by adding water (50 μL), and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (10 mg) as a white powder. MS (ESI) m/z: 475.1 [M+H]+
(2S)-3-methyl-2-[4-(3-methylisoxazol-5-yl)triazol-1-yl]butanoic acid (32 mg) described in Reference Example 153 and the (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride (23 mg) described in Reference Example 122 were dissolved in DMF (1 mL), HATU (60 mg) and DIPEA (100 μL) were added at room temperature and the mixture was stirred for 1.5 hr. After completion of the reaction, chloroform and water were added, and the organic layer was separated by a Phase-separator (registered trade mark) and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (18 mg) as a pale-yellow powder.
MS (ESI) m/z: 386.1 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-[[cyclopropanecarbonyl(4-phenylbutyl)amino]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (65 mg) described in Reference Example 156 was dissolved in methanol (0.9 mL), 2 M hydrochloric acid-methanol (456 μL) was added and the mixture was stirred for 18 hr. The reaction solution was concentrated under reduced pressure, and the obtained residue was added to 2 M hydrochloric acid-methanol (456 μL) at room temperature and the mixture was stirred for 5 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added, at room temperature, 1-fluorocyclopropanecarboxylic acid (4.7 mg), HATU (17 mg), DIPEA (26 μL), and DMF (0.30 mL), and the mixture was stirred for 15 hr. HATU (17 mg) and DIPEA (26 μL) were added and the mixture was stirred for 3 hr. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (7.0 mg) as a colorless powder. MS (ESI) m/z: 582.3 [M+H]+
tert-Butyl N-[(1S)-1-[(2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-2-[4-[[cyclopropylmethyl(4-phenylbutanoyl)amino]methyl]-1H-imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (50 mg) described in Reference Example 157 was dissolved in 2 M hydrochloric acid-methanol (886 μL) at room temperature, and the mixture was stirred for 15 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added, at room temperature, 1-fluorocyclopropanecarboxylic acid (3.5 mg), HATU (14 mg), DIPEA (21 μL), and DMF (0.24 mL), and the mixture was stirred for 3 hr was stirred. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (8.8 mg) as a colorless powder.
MS (ESI) m/z: 582.3 [M+H]+
DMF (0.70 mL) was added to 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8, N-(5-phenylpentyl)cyclopropanamine (11 mg) described in Reference Example 161, DIPEA (30 μL), and HATU (30 mg) were added, and the mixture was stirred at room temperature overnight. N-(5-phenylpentyl)cyclopropanamine (11 mg) described in Reference Example 161 and HATU (15 mg) were added, and the mixture was stirred for about 1 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (18 mg) as a white powder. MS (ESI) m/z: 564.4 [M+H]+
DMF (0.60 mL) was added to 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (20 mg) described in Reference Example 8, N-(2-benzyloxyethyl)cyclopropaneamine (12 mg) described in Reference Example 163, HATU (30 mg), and DIPEA (0.027 mL) were added, and the mixture was stirred at room temperature for about 3 hr. To the reaction mixture was added water and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and purified by NH silica gel column chromatography (ethyl acetate:methanol=100:0-90:10) to give the title compound (15 mg) as a white powder.
MS (ESI) m/z: 552.5 [M+H]+
DMF (0.80 mL) was added to 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid (16 mg) described in Reference Example 8, N-(4-phenylbutyl)cyclopropanamine (20 mg) described in Reference Example 160, DIPEA (27 μL), and HATU (30 mg) were added and the mixture was stirred at room temperature for about 4 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (4.6 mg) as a white powder. MS (ESI) m/z: 550.4 [M+H]+
tert-Butyl N-j[(1S)-1-[(2S,4R)-4-hydroxy-2-[1-methyl-5-(4-phenylpiperidine-1-carbonyl)imidazol-2-yl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate (52 mg) described in Reference Example 159 was dissolved in 1,4-dioxane (0.46 mL), 4 M hydrochloric acid-dioxane (687 μL) was added at room temperature and the mixture was stirred for 6 hr. After completion of the reaction, the reaction solution was concentrated under reduced pressure. To the obtained residue were added 1-fluorocyclopropanecarboxylic acid (3.4 mg), HATU (12 mg), DIPEA (19 μL), and DMF (0.44 mL) at room temperature, and the mixture was stirred for 15 hr. After completion of the reaction, the reaction mixture was diluted with DMSO, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (9.5 mg) as a colorless powder. MS (ESI) m/z: 554.3 [M+H]+
(2S,3S)-2-(4-ethoxytriazol-1-yl)-3-methylpentanoic acid (75 mg) described in Reference Example 155, (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride (25 mg) described in Reference Example 122, and HATU (100 mg) were dissolved in DMF (1 mL), DIPEA (200 μL) was added, and the mixture was stirred at room temperature for 1 hr. After completion of the reaction, to the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (12 mg) as a pale-brown powder. MS (ESI) m/z: 363.3 [M+H]+
tert-Butyl N-[(1S,2S)-1-[(2S,4R)-4-hydroxy-2-(1H-imidazol-2-yl)pyrrolidine-1-carbonyl]-2-methylbutyl]carbamate (41 mg) described in Reference Example 162 was dissolved in 1,4-dioxane (1.2 mL), 4 M hydrochloric acid-dioxane (0.9 mL) was added at room temperature and the mixture was stirred overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and azeotropically distilled with toluene. To the obtained residue were added, at room temperature, DMF (0.70 mL), DIPEA (0.038 mL), 2-(3-methylisoxazol-5-yl)acetic acid (10 mg), and HATU (32 mg) and the mixture was stirred for 2 hr. To the reaction mixture were added chloroform and water, and the organic layer was separated by a Phase-separator (registered trade mark), concentrated under reduced pressure, and purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (3.9 mg) as a white powder. MS (ESI) m/z: 390.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 6-bromo-1,2,3,4-tetrahydroisoquinoline.
MS (ESI) m/z: 572.0/573.9 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 7-bromo-1,2,3,4-tetrahydroisoquinoline.
MS (ESI) m/z: 572.0/573.9 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 1-phenylpiperazine. MS (ESI) m/z: 523.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 4-(4-piperidyl)pyridine. MS (ESI) m/z: 523.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 4-benzyloxypiperidine. MS (ESI) m/z: 552.2 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (23 mg), WSC·HCl (20 mg), and HOAt (15 mg) were dissolved in DMF (1.0 mL), 3-benzyloxypiperidine (30 mg) was added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (20 mg) as a white viscous substance.
MS (ESI) m/z: 552.2 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (23 mg), WSC·HCl (20 mg), and HOAt (15 mg) were dissolved in DMF (1.0 mL), 2-methyl-5-(4-piperidyl)pyridine (30 mg) was added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (17 mg) as a pale-yellow viscous substance.
MS (ESI) m/z: 537.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 5-methyl-2-(4-piperidyl)pyridine. MS (ESI) m/z: 537.2 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 1-[(2-fluorophenyl)methyl]piperazine.
MS (ESI) m/z: 555.2 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (26 mg), HATU (40 mg), and DIPEA (40 μL) were dissolved in DMF (1.0 mL), 2-phenylmorpholine (30 mg) was added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (21 mg) as a white viscous substance.
MS (ESI) m/z: 524.1 [M+H]+
DMF (0.60 mL) was added to 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (13 mg), DIPEA (0.028 mL), (3R,5S)-5-(4-methylthiazol-2-yl)pyrrolidin-3-ol (10 mg) described in Reference Example 164, and HATU (31 mg) were added, and the mixture was stirred at room temperature for 1 hr. To the reaction mixture was added water and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by NH silica gel column chromatography (ethyl acetate:methanol=100:0-80:20) to give the title compound (5.7 mg) as a colorless oil. MS (ESI) m/z: 366.0 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-phenylpiperidine-3-carboxamide. MS (ESI) m/z: 565.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-methyl-N-pyrrolidin-3-yl-benzamide hydrochloride.
MS (ESI) m/z: 565.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and N-phenylpiperidine-4-amine dihydrochloride.
MS (ESI) m/z: 537.5 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 4-(ethylaminomethyl)-N-methyl-benzamide hydrochloride described in Reference Example 165. MS (ESI) m/z: 553.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 4-(ethylaminomethyl)-N,N-dimethylbenzamide hydrochloride.
MS (ESI) m/z: 567.5 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (20 mg), WSC·HCl (15 mg), and HOAt (11 mg) were dissolved in DMF (1.0 mL), N-methyl-1-[4-(2-methyl-4-pyridyl)phenyl]methanamine dihydrochloride (40 mg) described in Reference Example 170 and DIPEA (0.10 mL) were added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (17 mg) as a white viscous substance. MS (ESI) m/z: 573.5 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (20 mg), WSC·HCl (15 mg), and HOAt (11 mg) were dissolved in DMF (1.0 mL), N-methyl-1-[4-(6-methyl-3-pyridyl)phenyl]methanamine dihydrochloride (35 mg) described in Reference Example 172 and DIPEA (0.10 mL) were added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (20 mg) as a white viscous substance. MS (ESI) m/z: 573.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1n-imidazole-4-carboxylic acid described in Reference Example 8 and 1-[1-[(2-fluorophenyl)methyl]-4-piperidyl]-N-methylmethanamine dihydrochloride described in Reference Example 173. MS (ESI) m/z: 597.5 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (20 mg), WSC·HCl (15 mg), and HOAt (11 mg) were dissolved in DMF (1.0 mL), N-methyl-1-(1-phenyl-4-piperidyl)methanamine dihydrochloride (40 mg) described in Reference Example 167 and DIPEA (0.10 mL) were added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (16 mg) as a white viscous substance. MS (ESI) m/z: 553.4 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 156 using 2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 and 1-benzylpiperazin-2-one. MS (ESI) m/z: 551.4 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (26 mg), HATU (40 mg), and DIPEA (40 μL) were dissolved in DMF (1.0 mL), N-methyl-N′-phenylethane-1,2-diamine dihydrochloride (30 mg) and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (8 mg) as a colorless oil.
MS (ESI) m/z: 511.1 [M+H]+
2-[(2S,4R)-4-hydroxy-1-[2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoyl]pyrrolidin-2-yl]-1H-imidazole-4-carboxylic acid described in Reference Example 8 (26 mg), HATU (40 mg), and DIPEA (40 μL) were dissolved in DMF (1.0 mL), (2R)—N-benzyl-2-(methylamino)propanamide hydrochloride (30 mg) was added and the mixture was stirred at room temperature overnight. MeOH (1 mL) was added and the mixture was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (5.5 mg) as a colorless oil.
MS (ESI) m/z: 553.2 [M+H]+
To a solution of ethyl 3-methyl-2-[4-(3-methylisoxazol-5-yl)pyrazol-1-yl]butanoate (16 mg) described in Reference Example 177 in methanol (2 mL) was added 1 M sodium hydroxide aqueous solution (0.17 mL), and the mixture was stirred at 50° C. for 2 hr. The mixture was neutralized with 1 M hydrochloric acid and concentrated under reduced pressure. To the obtained residue were added (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride (13 mg) described in Reference Example 122, DMF (2 mL), and DIPEA (50 μL), and the mixture was stirred at room temperature for 2 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with water, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol=100:0-90:10) to give the title compound (8.3 mg) as a white powder.
MS (ESI) m/z: 385.1 [M+H]+
The title compound was obtained by the same reaction and treatment as in Example 44 using (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride described in Reference Example 122 and 3-methyl-2-(5-propyl-1,2,4-oxadiazol-3-yl)butanoic acid described in Reference Example 178. MS (ESI) m/z: 348.1 [M+H]+
. The title compound was obtained by the same reaction and treatment as in Example 44 using (3R,5S)-5-(1H-imidazol-2-yl)pyrrolidin-3-ol hydrochloride described in Reference Example 122 and 3-methyl-2-(5-propyl-1,3,4-oxadiazol-2-yl)butanoic acid described in Reference Example 179. MS (ESI) m/z: 348.1 [M+H]+
To a solution of Reference Example compound 181 (40 mg) in DMF (1 mL) were added N-methylmorpholine (0.027 mL), HOBt (13 mg), and EDCI (24 mg) and the mixture was stirred at room temperature for 0.5 hr. Reference Example 182 (50 mg) was added and the mixture was further stirred for 0.5 hr. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (water (0.1% TFA)-acetonitrile) to give the title compound (76 mg) as a yellow solid.
MS (ESI) m/z: 1053.8 [M+H]+
To a mixture of Reference Example compound 181 (42 mg), DMF (2.0 mL), DIPEA (0.29 mL), and 2-(2-tertbutoxy-2-oxo-ethoxy)acetic acid (17 mg) was added HATU (33 mg) under ice-cooling and the mixture was stirred at room temperature for 1 hr. To the reaction mixture was added water, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography to give the title compound (39 mg) as a colorless oil. MS (ESI) m/z: 612.5 [M+H]+
To a solution of Example compound 164-1 (39 mg), DMF (1.5 mL), DIPEA (0.145 mL), and Reference Example compound 180 (54 mg) was added HATU (33 mg) under ice-cooling, and the mixture was stirred at room temperature for 3 hr. To the reaction mixture was added water and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. To a mixture of the obtained residue and chloroform (2 mL) was added TFA (1.0 mL) under ice-cooling and the mixture was stirred for 3 hr. The reaction mixture was diluted with chloroform and toluene, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give the title compound (6 mg) as a pale-yellow solid.
MS (ESI) m/z: 1155.7 [M+H]+
To a solution of Reference Example compound 183 (25 mg) in ethanol (2 mL) was added 10% Pd/C (100 mg), and the mixture was stirred under hydrogen atmosphere at 60° C. for 8 hr Insoluble material in the reaction mixture was filtered off through celite, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (10 mM ammonium carbonate aqueous solution-acetonitrile) to give a colorless oil (8 mg). Using the obtained oil, the title compound (14 mg) was obtained as a pale-yellow solid by the same reaction and treatment as in Example 164 (164-2).
MS (ESI) m/z: 1097.4 [M+H]+
As the device, Biacore T200 (manufactured by GE Healthcare) was used. VHL was obtained by co-expressing Elongin B and Elongin C with a Twin-Strep-tag added to the N-terminus. As the sensor chip, Sensor chip CM5 (manufactured by GE Healthcare) was used. First, Strep-Tactin XT (manufactured by GE Healthcare) was immobilized on the sensor chip by amine coupling. Thereafter, VHL was immobilized using the affinity of Strep-Tactin XT and Twin-Strep-tag, and VHL was further immobilized on the sensor chip by amine coupling.
As the running buffer, a mixed solution of 20 mM HEPES pH 7.5, 150 mM NaCl, 1 mM TCEP, 0.005% Tween 20, and 2% DMSO was used. Solutions of various compound concentrations of the test compounds (compounds described in Examples 1 to 162 above) were flowed on the sensor chip with VHL immobilized thereon at a flow rate of 30 uL/min under 25° C. conditions, and changes in mass on the sensor chip were observed. The Contact time and the Dissociation time were determined according to the properties of the compounds. For many compounds, 60 seconds and 120 seconds were respectively used. The observed data was analyzed using Biacore T200 Evaluation software Version 2.0 (manufactured by GE Healthcare) and the Kd value for VHL of each compound was calculated.
The test results of this Experiment are shown in Table 3 below. In the Table, “***” indicates a Kd value of less than 1 M “**” indicates a Kd value of not less than 1 μM and less than 10 μM, and “*” indicates a Kd value of not less than 10 μM and less than 50 μM.
Using Example compounds 164 and 165, the degradation-inducing action on WDR5 protein in cancer cells was evaluated using LNCaP cell line which is a cell line derived from human prostate cancer. As a control, GAPDH was used. The test results are shown in
Cell culture LNCaP cell line was seeded in an evaluation plate in DMEM culture medium containing 10% bovine serum (FBS). A test compound was dissolved in DMSO, a dilution series was prepared, mixed with a culture medium, added to an evaluation plate, and cultured for one day and one night to expose the is cells to the compound. After the above-mentioned culture, the cells were collected and lysed using a cell lysing solution (RIPA buffer) containing a protease inhibitor (Halt Protease Inhibitor Cock tailt). This cell lysate was dissolved in an electrophoresis buffer (NuPAGE LDS Sample Buffer, manufactured by ThermoFischer) added with a reducing agent (NuPAGE Sample Reducing Agent, manufactured by ThermoFischer). After a high temperature treatment (95° C., 5 min) of the lysate, NuPAGE electrophoresis was performed. Proteins in the NuPAGE gel were transferred to a PVDF membrane by using a semi-dry transfer device (iBlot2 system, manufactured by ThermoFischer). After the transfer, Western blotting was performed using iBind Western Systems (manufactured by ThermoFischer). In practice, after blocking with iBind Solution adjusted to lX, the membrane was placed in the iBind Western Device, primary antibody (anti-human WDR5 protein rabbit antibody, manufactured by Cell Signaling) solution, secondary antibody (anti-rabbit WDR5 protein labeled antibody, manufactured by Cell Signaling), and 1XiBind Solution were added to a designated place and allowed to stand at room temperature for 2 hr 30 min. The membrane after practice was washed with ultrapure water (water purified with a MilliQ device, manufactured by Merck Millipore), an antibody-dependent chemiluminescence was performed with a coloring solution for HRP (Immobilon Western Chemiluminescent Substrate, manufactured by Merck Millipore), and a band corresponding to WDR5 protein on the membrane was detected using an image analyzer (Amersham Imager 600, manufactured by Amersham).
Using Example compound 163, a Hibit assay was performed. The test results are shown in
A BRPF1 protein-expressing plasmid vector added with an 11-amino acid peptide tag Hibit was introduced into the cells by using an introduction reagent such as Lipofectamine and the like. Thereafter, a test compound was diluted with a culture medium and added. After culturing for a while, the amount of Hibit-added protein was quantified by measuring the luminescent intensity with a plate reader or the like using Nano-Glo HiBiT Lytic Detection System.
The present invention provides a target protein degradation-inducing compound that is a bifunctional compound having a portion that binds to Von-Hippel-Lindau, which is a substrate recognition protein of a ubiquitin ligase complex constituted of low-molecular-weight compounds, at one end, and a portion that is capable of binding or binds to a target protein at the other end.
This application is based on a patent application No. 2020-065410 filed in Japan (filing date: Mar. 31, 2020), the contents of which are incorporated in full herein.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2020-065410 | Mar 2020 | JP | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/JP2021/013694 | 3/30/2021 | WO |
