HK1127920A - 3-deazapurine derivatives as tlr7 modulators - Google Patents
3-deazapurine derivatives as tlr7 modulators Download PDFInfo
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Description
Technical Field
The invention relates to a 3-deazapurine derivative. The invention also relates to a preparation method of the 3-deazapurine derivative, an intermediate for preparing the 3-deazapurine derivative, a pharmaceutical composition containing the 3-deazapurine derivative and an application of the 3-deazapurine derivative.
Background
Toll-like receptors (TLRs) are major transmembrane proteins characterized by an extracellular leucine rich and cytoplasmic tail containing a conserved region known as the Toll/IL-1 receptor (TIR) domain. They are expressed predominantly on immune cells (e.g., dendritic cells, T lymphocytes, macrophages, monocytes, and natural killer cells), which play an important role in the innate immune system. They are a group of pattern recognition receptors that bind to pathogen-associated molecular patterns (for Reviews see, e.g., Ulevitch, R.J., Nature Reviews: Immunology, 4, 512-. Their name is derived from the homologous sequence of the Drosophila melanogaster (Drosophila melanogaster) gene Toll, which is found in Drosophila melanogaster and plays an important role in protecting Drosophila melanogaster from fungal infection (Hoffmann, J.A., Nature, 426, 33-38, 2003). 11 TLRs have been identified in mammals, and other non-mammalian TLRs are found in other vertebrates. All TLRs appear to function as homodimers or heterodimers in recognizing a specific or set of specific molecular determinants present on pathogenic organisms, including bacterial cell surface lipopolysaccharides, lipoproteins, bacterial flagellins, DNA from bacteria and viruses, and viral RNA. The cellular response to TLR activation involves activation of one or more transcription factors, leading to the production and secretion of cytokines and co-stimulatory molecules such as interferon, TNF- α, interleukins, MIP-1 and MCP-1, which act to kill and eliminate the invasion of pathogens.
Thus, there is an ongoing need to provide TLR7 modulators, particularly agonists. Preferably, such compounds should have one or more of the following properties: they should bind selectively to the TLR7 receptor; the gastrointestinal tract has good absorption; metabolically stable and have favorable pharmacokinetic properties; hardly shows side effects and is easy to prepare.
Disclosure of Invention
The present inventors have discovered a series of 3-deazapurine derivatives that are modulators, especially agonists, of the TLR7 receptor that find application in a variety of therapeutic areas involving modulation, especially agonism, of the TLR7 receptor, including the treatment of viral infections such as HCV or HBV, cancer and tumors, and diseases mediated by T2 helper cells (TH 2).
According to a first aspect of the present invention there is provided a compound of formula (I),
or a pharmaceutically acceptable salt or solvate of the compound, wherein
(a) Y is a bond, and R3Selected from aryl, (C)1-C6) Alkyl and- (C)1-C4) alkylene-O- (C)1-C4) An alkyl group; or
(b) Y is (C)1-4) Alkylene, and R3Selected from aryl, (C)3-C7) Cycloalkyl and 3-10 membered heterocyclyl;
z is oxygen or absent;
R1selected from H, halogen, OH, CN, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NHSO2R6、-NR6R7、-C(O)R6、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8Aryl and 3-10 membered heterocyclyl;
R2Selected from H, halogen, OH, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NR6R7、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8And a 3-10 membered heterocyclic group; or
R1And R2Can be connected to form (C)2-C5) An alkylene linkage, the linkage optionally comprising 1 or 2Each independently selected from N, O and S;
R5is absent and R4Selected from H, (C)3-C7) Cycloalkyl, aryl, - (CH)2) Aryl, -C (O) R9、-CO2R9、-(C1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9)、-C(O)NR9R10、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2(ii) a Or
R4Is absent and R5Is selected from R9、-C(O)R9、-CO2R9、-(C1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-C(O)NR9R10、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2；
R6And R7Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and- (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R6And R7Together with the nitrogen to which they are attached form a 3-6 membered saturated heterocyclic ring optionally additionally containing 1 or 2 heteroatoms selected from N, O and S;
R8is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and phenyl;
R9and R10Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, aryl, - (CH)2) Aryl and 3-10 membered heterocyclyl; or
R9And R10Together with the nitrogen to which they are attached form a 3-to 10-membered hetero ringA cyclic group;
R11and R12Independently selected from H and (C)1-C6) An alkyl group; or R11And R12Together with the attached N form a 3-6 membered saturated heterocyclyl, optionally additionally containing 1 or 2 heteroatoms selected from N, O and S;
The alkyl, cycloalkyl, alkoxy, aryl and heterocyclyl groups are optionally substituted with one or more atoms or groups independently selected from: halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12、-(C1-C6) alkylene-NR11R12Aryl and 3-10 membered heterocyclyl;
with the proviso that if R1And R2Is H and Z and R5If not, then:
(a) when Y-R3When it is ethyl, R4Is not methyl; and is
(b) When Y-R3When it is methyl, R4Not H or methyl.
Unless otherwise specified, alkyl and alkoxy groups may be straight or branched chain groups and contain 1 to 6 carbon atoms, and preferably 1 to 4 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, pentyl and hexyl. Examples of alkoxy groups include methoxy, ethoxy, isopropoxy, and n-butoxy.
Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and bicycloheptyl.
Halogen means fluorine, chlorine, bromine or iodine, and is preferably fluorine or chlorine.
Aryl groups include phenyl, naphthyl, anthryl and phenanthryl, and are preferably phenyl.
Unless otherwise specified, the heterocyclic ring may be saturated, partially saturated, or aromatic and contain one or more heteroatoms independently selected from N, O and S. For example, the heterocyclic ring can be a 5-6 membered saturated, partially saturated, or aromatic heterocyclic ring. Examples of saturated heterocyclic groups are: tetrahydrofuranyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, dioxolanyl, dihydropyranyl, tetrahydropyranyl, piperidinyl, pyrazolinyl, dioxanyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, azepinyl, oxazepinyl, and thiazepinyl. Examples of aromatic monoheterocyclic groups are: pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl (such as 1, 2, 3-triazolyl and 1, 2, 4-triazolyl), oxadiazolyl (such as 1-oxa-2, 3-oxadiazolyl, 1-oxa-2, 4-oxadiazolyl, 1-oxa-2, 5-oxadiazolyl and 1-oxa-3, 4-oxadiazolyl), thiadiazolyl (such as 1-thia-2, 3-oxadiazolyl, 1-thia-2, 4-oxadiazolyl, 1-thia-2, 5-oxadiazolyl and 1-thia-3, 4-oxadiazolyl), tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl and triazinyl. Examples of bicyclic aromatic heterocyclic groups are: benzofuranyl, benzothienyl, indolyl, benzimidazolyl, indazolyl, benzotriazolyl, quinolinyl, and isoquinolinyl groups.
When multiple substituents can be selected from a large number of substituent groups, the selected groups can be the same or different.
In one aspect, Z is oxygen, so as to form an N-oxide.
In another embodiment of the invention, Z is absent.
In still another embodiment of the present invention, R1Is selected from
(a)H；
(b)CN；
(c) Halogen;
(d) optionally substituted by 1 to 3 halogensAtom substituted (C)1-C6) An alkyl group;
(e) tetrahydrofuryloxy;
(f) substituted with a 3-6 membered saturated heterocyclic group containing 1-3 heteroatoms independently selected from N, O and S (C)1-C6) Alkyl, wherein the heterocyclyl is optionally substituted with 1-3 substituents independently selected from CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy and- (C)1-C6) alkylene-O- (C)1-C6) Substituted with a group in alkyl;
(g)-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
(h)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(i) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(j)(C3-C7) A cycloalkyl group;
(k)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(1)-C(O)NR6R7；
(m)-CO2R6；
(n)-C(O)R6；
(o) a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, said 5-and 6-membered aromatic heterocyclic groups optionally being substituted with 1 to 3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C) 1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(p) phenyl optionally substituted with 1 to 3 halogen atoms;
(q)-NR6R7；
(r)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
(s) or R1And R2Can be connected to form (C)2-C5) An alkylene linkage;
wherein
R6、R7、R11And R12As defined in the first aspect of the invention.
In another embodiment, R1Is selected from
(a)H；
(b)CN；
(c) Halogen element
(d) Optionally substituted by 1 to 3 halogen atoms1-C6) An alkyl group;
(e) tetrahydrofuryloxy;
(f) substituted by morpholinyl, piperazinyl or pyrrolidinyl optionally substituted by 1 or 2 methyl groups1-C6) An alkyl group;
(h)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(i) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(j)(C3-C7) A cycloalkyl group;
(k)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(l)-C(O)NR6R7；
(m)-CO2R6；
(n)-C(O)R6；
(o) a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, said 5-and 6-membered aromatic heterocyclic groups being optionally substituted with 1 to 3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(p) phenyl optionally substituted with 1 to 3 halogen atoms;
(q)-NR6R7；
(r)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
wherein
R6、R7、R11And R12As defined in the first aspect of the invention.
In yet another embodiment, R1Selected from (C) optionally substituted by 1 to 3 halogen atoms1-C4) An alkyl group; (C)3-C7) A cycloalkyl group; or optionally substituted with 1-3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl and-NH (C)1-C6) Atoms or radicals in alkyl radicalsA substituted 5-6 membered aromatic heterocyclic group.
In yet another embodiment, R1Selected from methyl or ethyl substituted with 1-3 fluorine atoms; a cyclopropyl group; - (C)1-C2) alkylene-O- (C)1-C2) An alkyl group; optionally substituted by OH or cyclopropyl (C)1-C4) An alkoxy group; -COCH3；-CH2OCH3(ii) a and-CO2CH3。
In yet another embodiment, R1Is cyclopropyl or CF3。
In yet another embodiment, R1Is a 5-membered aromatic heterocyclic group comprising: (i)1-4 nitrogen atoms, or (ii)1-2 nitrogen atoms and 1 oxygen or sulfur atom, or (iii)1 oxygen or sulfur atom, said 5-membered aromatic heterocyclic group being optionally substituted with 1-3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C3) alkylene-O- (C)1-C4) Alkyl, - (C)1-C4) alkylene-OH, -R11R12And- (C)1-C3) alkylene-NR11R12Wherein R is substituted with an atom or group of (1)11And R12As defined in the first aspect of the invention.
In yet another embodiment, R1Selected from imidazolyl, oxazolyl, oxadiazolyl, triazolyl, pyrazolyl and thiazolyl, wherein each of these groups is optionally selected from 1-3 independently halogen, OH, CF 3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C3) alkylene-O- (C)1-C4) Alkyl, - (C)1-C4) alkylene-OH and- (C)1-C3) alkylene-NR11R12Wherein R is substituted with an atom or group of (1)11And R12As defined in the first aspect of the invention.
In yet another embodiment, R1Selected from unsubstituted oxazolyl, triazolyl,Pyrazolyl and thiazolyl.
In yet another embodiment, R1Is oxazolyl.
In yet another embodiment, R2Is selected from
(a)H；
(b) Halogen;
(c) optionally substituted by 1 to 3 halogen atoms1-C6) An alkyl group;
(d) tetrahydrofuryloxy;
(e) substituted with a 3-6 membered saturated heterocyclic group containing 1-3 heteroatoms independently selected from N, O and S (C)1-C6) Alkyl, wherein the heterocyclyl is optionally substituted with 1-3 substituents independently selected from CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy and- (C)1-C6) alkylene-O- (C)1-C6) Substituted with a group in alkyl;
(f)-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
(g)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(h) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(i)(C3-C7) A cycloalkyl group;
(j)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(k)-C(O)NR6R7；
(1)-CO2R6；
(m)-C(O)R6；
(n) containing (i)1 to 4 nitrogen atomsOr (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, said 5-and 6-membered aromatic heterocyclic groups being optionally substituted with 1 to 3 substituents independently selected from halogen, OH, CF 3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(o) phenyl optionally substituted with 1 to 3 halogen atoms;
(p)-NR6R7；
(q)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
wherein
R6、R7、R11And R12As defined in the first aspect of the invention.
In yet another embodiment, R2Is H or methyl.
In yet another embodiment, R2Is H.
In yet another embodiment, Y is methylene; and R is3Is aryl or a 5-6 membered heterocyclyl, said heterocyclyl containing 1-3 heteroatoms independently selected from N, O and S, said aryl and heterocyclyl being optionally substituted with 1-3 atoms or groups independently selected from: halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, -NH (C)1-C6) Alkyl, -N ((C)1-C6) Alkyl radical)2Aryl and 3-10 membered heterocyclyl.
In yet another embodiment, Y is methylene; and R is3Selected from: an aryl group, a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom, and a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms; the aryl and aromatic heterocycle are optionally substituted with 1-3 atoms or groups independently selected from: halogen, OH, oxo, CF 3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12、-(C1-C6) alkylene-NR11R12Aryl and 3-10 membered heterocyclyl, wherein R11And R12As defined in the first aspect of the invention.
In yet another embodiment, Y is methylene; and R is3Selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, each of which is optionally substituted with 1-3 substituents independently selected from halogen, (C)1-4) Alkyl, (C)1-C4) Alkoxy and CF3Is substituted with the atom or group in (1).
In yet another embodiment, Y is methylene; and R is3Selected from phenyl, pyridin-3-yl and 6-methyl-pyridin-3-yl.
In a further aspect of the present invention,
y is methylene;
R1selected from: substituted by 1 to 3 halogen atoms1-C4) An alkyl group; (C)3-C7) A cycloalkyl group; and a 5-6 membered aromatic heterocyclic group optionally substituted with 1 to 3 groups independently selected from: halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl and-NH (C)1-C6) An alkyl group;
R2is H;
R3is phenyl or 3-pyridyl, each of which is optionally substituted by 1 to 3 substituents independently selected from halogen, (C)1-4) Alkyl and CF3Is substituted with the atom or group in (1);
R4is H; and is
R5Is absent.
In a further aspect of the present invention,
y is methylene;
R1selected from CF3Cyclopropyl and oxazolyl;
R2Is H;
R3selected from phenyl, pyridin-3-yl and 6-methyl-pyridin-3-yl.
R4Is H; and is
R5Is absent.
In yet another embodiment, R5Is absent; and is
R4Is selected from- (C)1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2Wherein Y, Z, R1、R2、R3、R9And R10As defined in the first aspect of the invention, to produce a compound of formula (Ia) as follows:
in yet another embodiment, R4Is H and R5Is absent.
In yet another embodiment, R4Is absent; and is
R5Is selected from- (C)1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2Wherein Y, R1、R2、R3、R9And R10As defined above, to yield a compound of formula (Ib) as follows:
embodiments of the invention form a further aspect of the invention.
Preferred compounds of the invention are those of examples 1-4, 12, 15-18, 26, 27, 36-38, 40-54, 60, 70, 76, 78, 82, 83, 86, 92-94, 96-98 and 100 and tautomers thereof and pharmaceutically acceptable salts or solvates of the compounds or tautomers thereof.
More preferred compounds are selected from:
4-amino-1-benzyl-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one (example 1);
4-amino-1-benzyl-6-oxazol-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one (example 12);
4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one (example 15);
and tautomers thereof, and pharmaceutically acceptable salts or solvates of the compounds or tautomers thereof.
In another embodiment of the present invention, there are provided compounds of formula (Ic)
Wherein
Y is methylene;
R1and R2Each independently selected from H, halogen, OH, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NR6R7、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8Aryl and 3-10 membered heterocyclyl; or
R1And R2Can be connected to form (C)2-C5) An alkylene linkage, the linkage optionally comprising 1 or 2 heteroatoms each independently selected from N, O and S;
R3is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, aryl, and 3-10 membered heterocyclyl;
R4is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10And R is5Is absent; or
R5Is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10And R is4Is absent;
R6and R7Each independently selected from H and (C)1-C6) An alkyl group;
R8is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and phenyl;
R9and R10Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl radicalsAryl, - (CH)2) Aryl and 3-10 membered heterocyclyl; or
R9And R10Together with the nitrogen to which they are attached form a 3-10 membered heterocyclyl;
the alkyl, cycloalkyl, alkoxy, aryl and heterocyclyl groups are optionally substituted with one or more groups independently selected from: halogen, OH, oxo, CF 3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy group, (C)1-C6) Alkoxy (C)1-C6) Alkyl, NH (C)1-C6) Alkyl, N ((C)1-C6) Alkyl radical)2Aryl and 3-10 membered heterocyclyl;
or a pharmaceutically acceptable salt or solvate of the compound;
with the proviso that if R1And R2Is H and z and R5Is absent, then
(a) If Y-R3Is ethyl, R4Is not methyl; or
(b) If Y-R3Is methyl, R4Not H or methyl.
Yet another embodiment of the present invention includes compounds of formula (Ic), wherein R is4Is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10；R5Is absent; and Y, R1、R2、R3、R9And R10As defined in the second aspect of the invention, to produce a compound of formula (Ia) as shown hereinbefore.
Yet another embodiment of the present invention includes compounds of formula (Ic), wherein
Y is methylene;
R1and R2Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Ring (C)Alkyl, -CO2H、-CO2(C1-C6) Alkyl and-C (O) NH (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R1And R2Can be connected to form (C)2-C5) An alkylene linkage;
R3is phenyl, optionally substituted with one or more groups independently selected from: halogen, OH, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, NH (C)1-C6) Alkyl and N ((C)1-C6) Alkyl radical)2；
R5Is absent; and is
R4Is H.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein
Y is methylene;
R1And R2Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, -CO2H、-CO2(C1-C6) Alkyl and-C (O) NH (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R1And R2Can be connected to form (C)2-C5) An alkylene linkage;
R3is phenyl, optionally substituted by one or more substituents independently selected from halogen and CF3Substituted with the group (1);
R5is absent; and is
R4Is H.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is5Is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10；R4Is absent; and Y, R1、R2And R3As defined above, to yield a compound of formula (Ib) as shown above.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is1And R2Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, -CO2R6、-C(O)NR6R7and-C (O) NR6SO2R8(ii) a Or R1And R2Can be connected to form (C)2-C5) An alkylene linkage optionally containing 1 or 2 heteroatoms each independently selected from N, O and S.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is1And R2Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, -CO2H、-CO2(C1-C6) Alkyl and-C (O) NH (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R1And R2Can be connected to form (C)2-C5) An alkylene linkage.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is 1And R2Each independently selected from H, (C)1-C3) Alkyl, cyclopropyl, -CO2H、-CO2CH3and-C (O) NH (CH)2) A cyclopropyl group; or R1And R2Can be connected to form C5-an alkylene linkage.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is1Selected from H, methyl, n-propyl, isopropyl, cyclopropyl, -CO2H、-CO2CH3and-C (O) NH (CH)2) A cyclopropyl group; and R is2Selected from H and methyl; or R1And R2Can be connected to form C5-an alkylene linkage.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is3Is aryl, optionally substituted with one or more groups independently selected from: halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy group, (C)1-C6) Alkoxy (C)1-C6) Alkyl, NH (C)1-C6) Alkyl, N ((C)1-C6) Alkyl radical)2Aryl and 3-10 membered heterocyclyl.
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is3Is phenyl, optionally substituted with one or more groups independently selected from: halogen, OH, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, NH (C)1-C6) Alkyl and N ((C)1-C6) Alkyl radical)2. However, R is more preferred3Is optionally selected from one or more of halogen and CF3Phenyl substituted with the group of (1).
Yet another embodiment of the second aspect of the invention includes compounds of formula (Ic), wherein R is 3Selected from the group consisting of 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl and 3-trifluoromethylphenyl.
Unless otherwise indicated, the compounds of the present invention include compounds of formulae (I), (Ia), (Ib) and (Ic).
It is to be understood that the present invention encompasses all combinations of the embodiments of the invention described above which are consistent with the definition of the compound of formula (I).
In a third aspect of the invention, there is provided a process for the preparation of a compound of formula (Ic)
Wherein, in the formulae Ia, XVIII, XVIIa, XIX, XIXa, XXa, XXb, XIV, XV, LIV and LXIII, Y-R3As defined in claim 14, R1As defined in claim 2, R2As defined in claim 10, PG1And PG2Is a nitrogen protecting agent and R13Is (C)1-6) An alkyl group;
the method comprises the following steps:
(a) reacting a compound of formula (XVIII) or (XVIIIa) with a carbonyl donor
To form the corresponding compound of formula (XIX) or (XIXa),
then deprotecting the compound of formula (XIX) or (XIXa); or
(b) Reduction of a compound of formula (XXa)
To form a compound of formula (XXb)
The compound of formula (XXb) is then cyclized by treatment with a protic acid; or
(c) Reduction of a compound of formula (XIV)
To form a compound of formula (XV)
Then cyclizing the compound of formula (XV) in the presence of a carbonyl moiety; or
(e) Cyclisation of a compound of formula (LIV) to the corresponding compound of formula (XIXA) hereinbefore in the presence of diphenylphosphonyl azide and deprotection of the amino protecting group
(ii) a Or
(f) Hydrolyzing the compound of formula (LXIII)
In a fourth aspect of the invention, there are provided intermediates of formula XVIII, XVIIa, XIX, XIXa, XXa, XXb, XIV, XV, LIV and LXIII,
wherein Y-R3As defined in claim 14, R1As defined in claim 2, R2As defined in claim 10, PG1And PG2Is a nitrogen protecting agent and R3Is (C)1-6) An alkyl group.
Pharmaceutically acceptable salts of the compounds of formula (I) include acid addition and base salts thereof.
Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include ethylhydrochloric acid, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hydroxybenzylbenzoate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malic acid, maleate, malonate, methanesulfonate, methylsulfate, naphthenate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/monohydrogen phosphate/dihydrogen phosphate, pyroglutamate, dihydrogenphosphate, and mixtures thereof, Sugar acid salts, stearates, succinates, tannates, tartrates, tosylates, trifluoroacetates, and xinofoate salts.
Suitable base salts are formed from bases which form non-toxic salts. Examples include aluminum salts, arginine salts, benzathine salts (benzathine), calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts (meglumine), ethanolamine salts (olamine), potassium salts, sodium salts, tromethamine salts (tromethamine), and zinc salts.
Hemisalts of acids and bases, such as hemisulfate and hemicalcium salts, may also be formed.
For an overview of suitable Salts, see Stahl and Wermuth, "Handbook of pharmaceutical Salts: properties, Selection, and use "(Wiley-VCH, Weinheim, Germany, 2002).
Pharmaceutically acceptable salts of the compounds of formula (I) may be prepared by one or more of the following three methods:
(i) reacting a compound of formula (I) with a desired acid or base;
(ii) removing the labile protecting group of an acid or base from a suitable precursor of a compound of formula (I) with the desired acid or base; or
(iii) One salt of the compound of formula (I) is converted to another salt by reaction with a suitable acid or base or by use of a suitable ion exchange column.
All three reactions are usually carried out in solution. The salt produced can be precipitated out of the river and collected by filtration or can be recovered by evaporation of the solvent. The resulting salts may be ionized to varying degrees (from fully ionized to nearly non-ionized).
The compounds of the present invention may also exist in unsolvated forms as well as solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). The term "hydrate" system is used when the solvent is water.
The currently accepted classification system for organic hydrates is the definition of separation sites (definesolated sites) defining separation sites, channels or metal ion coordinated hydrates, see k.r. morrisPolymorphism in Pharmaceutical Solids(H.g. Brittain eds., Marcel Dekker, 1995), which is incorporated herein by reference. The water molecules are separated by the insertion of organic molecules in the hydrates at the separation sites, without direct contact with each other. In channel hydrates, a water molecule is in a lattice channel where it is in close proximity to other water molecules. In the metal ion complex hydrate, water molecules are bonded to the metal ions.
When solvents or water are intimately combined, the complex has a well-defined stoichiometry that is independent of humidity. However, when the solvent or water binding is weak in the channel solvate and hygroscopic compound, the water/solvent content will depend on the humidity and drying conditions. In this case, the non-stoichiometric method is a standard amount.
The compounds of the present invention may exist in a continuous solid state ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a material that lacks long-range order at the molecular level and exhibits the physical characteristics of a solid or liquid depending on temperature. Typically these materials do not exhibit the characteristic X-ray diffraction pattern, and while exhibiting solid properties, are more often described as liquids. The transition from a solid to a liquid characteristic upon heating is then characterized by a change in state, typically two-stage ("glass transition"). The term "crystalline" refers to a solid state material having a regular ordered internal structure at the molecular level, which exhibits a characteristic X-ray diffraction pattern with defined peak states. These materials also exhibit liquid properties when heated sufficiently, but the change from a solid to a liquid state is characterized by a phase change, typically first order ("melting point").
The compounds of the invention may also be present as multicomponent complexes (other than salts and solvates) in which the drug and at least one of the other components are present in stoichiometric or non-stoichiometric amounts. This type of complex includes a caged compound (drug-host complex) and a co-crystal. The latter is generally defined as a crystalline complex of neutral molecular components that are bonded to each other by non-covalent bonds, but may also be a complex of a neutral molecule and a salt. Cocrystals can be prepared by melt crystallization, recrystallization from solvents, or by physically grinding the individual components together, see ChemCommun, almarson and m.j.zawortko, 171889-1896(2004), which is incorporated herein by reference. For a review of multicomponent complexes, see J PharmSci by Haleblian,64(8) 1269-1288(1975, 8), which is incorporated herein by reference.
The compounds of the invention may also be present in a mesomorphic state (mesophase or liquid crystal) when subjected to suitable conditions. A mesomorphic state is an intermediate state between a true crystalline state and a true liquid state (melt or solution). The mesogenic phenomena caused by temperature changes are referred to as "thermally induced" mesogenicity, and the mesogenic phenomena caused by the addition of a second component (e.g., water or other solvent) are referred to as "lyotropic" mesogenicity. Compounds with a tendency to form lyotropic mesophases are called "amphipathic molecules" and consist of molecules having ions (e.g. -COO)-Na+、-COO-K+or-SO3 -Na+) Or non-ionic (e.g. -N)-N+(CH3)3) The polar head group of (a). For more information please refer toN.H.Hartshorn and A.Stuart, crystalls and the Polarizing Microscope, 4 th edition (EdwardAlnold, 1970), which is incorporated herein by reference.
The compounds of formula (I) referred to hereinafter include their salts, solvates, multicomponent complexes and liquid crystals, and their solvates, multicomponent complexes and liquid crystals.
The compounds of the invention include compounds of formula (I) as hereinbefore defined, including all polymorphs and crystal habit thereof, prodrugs and isomers thereof (including optical, geometric and tautomeric isomers) and isotopically labelled compounds of formula (I) as hereinafter defined.
As mentioned above, so-called "prodrugs" of the compounds of formula (I) are also included within the scope of the present invention. Thus, certain derivatives of the compounds of formula (I) which themselves have little or no pharmacological activity are capable of being converted to compounds of formula (I) having the desired activity when administered in vivo or in vivo, for example by hydrolytic cleavage. These derivatives are referred to as "prodrugs". Further information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems" volume 14, ACS Symposium Series (T.Higuchi and W.Stella) and "Bioreperable Carriers in Drug Design", Pergamon Press, 1987(E.B.Roche eds., American Pharmaceutical Association).
Prodrugs of the invention may be prepared, for example, as a "precursor moiety" by substituting appropriate functional groups present in the compounds of formula (I) with moieties known to those skilled in the art, as described in H.Bundgaard "Design of produgs" (Elsevier, 1985).
Some examples of prodrugs of the invention include
(i) Wherein the compound of formula (I) contains a carboxylic acid function, an ester thereof, for example wherein the hydrogen in the carboxylic acid function of the compound of formula (I) is substituted with (C)1-C8) A compound of an alkyl group; and
(ii) wherein the compound of formula (I) contains a primary or secondary amine functional group, an amide group thereof, for example wherein 1 or 2 hydrogens of the amine functional group of the compound of formula (I) are substituted as (C) as the case may be1-C10) An alkanoyl group.
Further examples of substituents according to the above examples and examples of other prodrug types can be found in the aforementioned references. Furthermore, some compounds of formula (I) may themselves act as prodrugs of other compounds of formula (I).
In particular, R of the invention4As defined herein and is not H, and R5Compounds of formula (I) which are absent (i.e. compounds of formula (Ia)) may be converted to R by metabolic or solvolytic action4Is H and R5Absent compound of formula (I). Furthermore, R of the present invention4Is absent and R5Compounds of formula (I) as defined herein and which are not H (i.e. compounds of formula (Ib)) may be converted to R by metabolic or solvolytic action4Is absent and R5A compound of formula (I) which is H.
Metabolites of the compounds of formula (I) are also included within the scope of the present invention, i.e. compounds which are formed in vivo following administration of the drug. Some examples of metabolites according to the invention include
(i) When the compound of formula (I) contains a methyl group, it is a hydroxymethyl derivative (-CH) thereof3->-CH2OH)；
(ii) When the compound of formula (I) contains an alkoxy group, it is a hydroxy derivative thereof (-OR- > -OH);
(iii) when the compound of formula (I) contains a tertiary amino group, it is a secondary amino derivative thereof (-NR)1R2->-NHR1or-NHR2)；
(iv) When the compound of formula (I) contains a secondary amino group, it is a primary amino derivative thereof (-NHR)1->-NH2)；
(v) When the compound of formula (I) contains a phenyl moiety, it is a phenol derivative thereof (-Ph > -PhOH);
(vi) when the compound of formula (I) contains an amide group, it is a carboxylic acid derivative thereof (-CONH)2->-COOH)。
The compounds of formula (I) containing one or more asymmetric carbon atoms may exist as two or more stereoisomers. When the compound of formula (I) contains an alkenyl or alkenylene group, it may be a cis/trans (or Z/E) geometric isomer. Tautomerism ("tautomerism") can occur when structural isomers can interconvert with a low energy barrier. In compounds of formula (I) containing, for example, a keto group, it may take the proton tautomeric form, or in compounds containing an aromatic moiety, it may take the so-called valence tautomeric form. It follows that a single compound may exhibit more than one type of isomerism.
By way of example, the dotted line in formula (I) is illustrated, wherein R4A compound of formula (Ia) which is H is wherein R5A tautomer of a compound of formula (Ib) that is H:
all stereoisomers, geometric isomers and tautomeric forms of the compounds of formula (I), including compounds exhibiting more than one type of isomerism, and mixtures of more than one thereof, are included within the scope of the invention. The invention also includes acid addition salts in which the counterion is optically active (e.g., d-lactate or l-lysine) or racemic (e.g., dl-tartrate or dl-arginine).
The cis/trans isomers can be separated by conventional techniques known to those skilled in the art, such as chromatography and fractional crystallization.
Conventional techniques for the preparation/separation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of racemic compounds (or racemic compounds as salts or derivatives) using, for example, chiral High Performance Liquid Chromatography (HPLC).
Alternatively, the racemic compound (or racemic precursor) may be reacted with a suitable optically active compound (e.g. an alcohol; or a base or acid (when the compound of formula (I) contains an acidic or basic moiety), such as 1-phenylethylamine or tartaric acid). The resulting diastereomer mixture can be separated by chromatography and/or fractional crystallization, and 1 or 2 of the diastereomers can be converted to the corresponding pure enantiomers by methods well known to those skilled in the art.
The chiral compounds of the invention (and chiral precursors thereof) can be obtained in enantiomerically enriched form using chromatography (typically HPLC) on heterogeneous resins with a mobile phase consisting of a hydrocarbon, typically heptane or hexane containing 0-50% by volume (typically 2-20% by volume) isopropanol and 0-5% by volume of an alkylamine (typically 0.1% diethylamine). The eluate is concentrated to produce a concentrated mixture.
The present invention includes all crystalline forms of the compound of formula (I), including racemic compounds and racemic mixtures thereof. Stereoisomeric racemic mixtures can be isolated by conventional techniques known to those skilled in the art, see, for example, "Stereochemistry of Organic Compounds" (Wiley, New York, 1994) by E.L. Eliel and S.H. Wilen.
The present invention includes all pharmaceutically acceptable isotopically-labelled compounds of formula (I) wherein one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number preferred in nature.
Examples of isotopes suitable for inclusion in compounds of the invention include: isotopes of hydrogen, such as2H and3h; isotopes of carbon, such as 11C、13C and14c; isotopes of chlorine, such as36Cl; isotopes of fluorine, such as18F; isotopes of iodine, such as123I and125i; isotopes of nitrogen, such as13N and15n; isotopes of oxygen, such as15O、17O and18o; isotopes of phosphorus, such as32P; and isotopes of sulfur, such as35S。
Some isotopically-labeled compounds of formula (I) (e.g., those incorporating a radioisotope) are useful in drug and/or substrate tissue distribution studies. The radioactive isotope tritium (i.e.,3H) and carbon 14 (i.e.,14C) it is particularly suitable for this purpose because of its ease of incorporation and simplicity of the detection method.
Heavy isotopes (such as deuterium, i.e.2H) Substitution may be preferred in some instances because of the greater metabolic stability that may provide some therapeutic advantage (e.g., increased in vivo half-life or reduced dosage requirements).
Via positron emitting isotopes (such as11C、18F、15O and13n) substitution, can be used in Positron Emission Tomography (PET) studies to investigate substrate receptor occupancy.
Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques well known to those skilled in the art or by processes analogous to those described in the examples and preparations herein, using a suitable isotopically-labelled reagent in place of the non-labelled reagent conventionally used.
Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted (e.g., D)2O、d6-acetone, d6-DMSO) substituted.
Also included within the scope of the present invention are intermediate compounds, all salts, solvates and complexes thereof, as defined below, and all solvates and complexes of salts thereof, as defined above for compounds of formula (I). The present invention includes all polymorphs and crystal habit of the above mentioned substance types.
When preparing the compounds of formula (I) according to the invention, it will be obvious to the person skilled in the art that the intermediate forms which provide the best combination of features for the purposes of the preparation are routinely selected. These characteristics include melting point, solubility, processability and yield of the intermediate form, and the ease with which the purified product can be isolated.
The compounds of the invention intended for use as medicaments may be administered as crystalline or amorphous products, or may exist as solid continuous bodies (from completely amorphous to completely crystalline). They may be obtained, for example, in the form of solid fillers, powders or films by processes such as precipitation, crystallization, freeze drying, spray drying or evaporation drying. Microwave drying or high-frequency drying can also be used for this purpose.
They may be administered alone or in combination with one or more other compounds of the invention or in combination with one or more other drugs (or as any combination thereof). Typically, they will be administered as a formulation in combination with one or more pharmaceutically acceptable excipients. The term "excipient" is used herein to describe any ingredient other than a compound of the invention. The choice of excipients depends largely on the following factors: such as the particular mode of administration, the effect of excipients on solubility and stability, and the nature of the dosage form.
Pharmaceutical compositions suitable for delivery of the compounds of the present invention and methods for their preparation will be apparent to those skilled in the art. Such compositions and methods of preparation are described, for example, in Remington's Pharmaceutical Sciences, 19 th edition (Mack publishing company, 1995).
Suitable modes of administration include oral, parenteral, topical, inhalation/intranasal, rectal/vaginal and ocular/otic.
The compounds of the invention may be administered orally. Oral administration may include swallowing, so that the compound enters the gastrointestinal tract; or may be administered buccally or sublingually so that the compound enters the bloodstream directly from the mouth. Formulations suitable for oral administration include: solid formulations such as tablets, capsules containing particles, liquids or powders, lozenges (including liquid filled), chewables, multiparticulates and nanoparticles, gels, solid solutions, liposomes, films, pills (ova), sprays, liquid formulations and buccal/mucoadhesive patches.
Liquid preparations include suspensions, solutions, syrups and elixirs. Such formulations may be employed as fillers in soft or hard capsules and typically comprise a carrier (for example, water, ethanol, polyethylene glycol, propylene glycol, methyl cellulose, or a suitable oil) and one or more emulsifying agents and/or suspending agents. Liquid formulations may also be prepared by, for example, reconstituting the solid from the sachet.
The compounds of the present invention are also useful in fast acting, fast disintegrating agents such as those described by Liang and Chen in Expert Opinion in Therapeutic Patents, 11(6), 981-.
For a tablet dosage form, the drug may comprise from 1% to 80% by weight of the dosage form, preferably from 5% to 60% by weight of the dosage form, depending on the dosage. In addition to the drug, tablets typically contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch, and sodium alginate. Typically, the disintegrant comprises from 1 to 25% by weight of the dosage form, preferably from 5 to 20% by weight of the dosage form.
Binders are typically used to provide cohesiveness to the tablet formulation. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropylmethyl cellulose. Tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrous lactose and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and dibasic calcium phosphate dihydrate.
The tablets optionally may also contain surfactants such as sodium lauryl sulfate and polysorbate 80 and glidants such as silicon dioxide and talc. When included, the surfactant may comprise 0.2% to 5% by weight of the tablet and the glidant may comprise 0.2% to 1% by weight of the tablet.
Tablets typically also contain lubricating agents such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. The lubricant is typically present in an amount of 0.25 to 10% by weight of the tablet, preferably 0.5 to 3% by weight. Other possible ingredients include antioxidants, coloring agents, flavoring agents, preservatives, and taste masking agents.
Exemplary tablets contain up to about 80% by weight drug, about 10% to about 90% by weight binder, about 0% to about 85% by weight diluent, about 2% to about 10% by weight disintegrant, and about 0.25% to about 10% by weight lubricant. The tablet blend may be compressed directly or via a roller to form a tablet. Alternatively, the tablet blend or a portion of the blend may be wet, dry or melt granulated, melt congealed or extruded prior to tableting. The final formulation may comprise one or more layers, which may be coated or uncoated; and may even be encapsulated. A discussion of the preparation of tablets is found in "Pharmaceutical DosageForms" by h.lieberman and l.lachman: tablets ", Vol.1 (Marcel Dekker, New York, 1980).
Orally consumable films for human or veterinary use are typically pliable water-soluble film dosage forms or water-swellable film dosage forms which are rapidly dissolving or mucoadhesive and typically comprise a compound of formula (I), a film-forming polymer, a binder, a solvent, a humectant, a plasticizer, a stabilizer or emulsifier, a viscosity modifier, and a solvent. Some components of the preparation have more than one function.
The compounds of formula (I) may be water soluble or insoluble. The water soluble compound typically comprises from 1 wt% to 80 wt%, more typically from 20 wt% to 50 wt% solute. The poorly soluble compounds may constitute a significant proportion of the composition, typically up to 88% by weight of the solute. Alternatively, the compound of formula (I) may be in the form of multiparticulate microbeads.
The film-forming polymer may be selected from natural polysaccharides, proteins or synthetic hydrocolloids and is typically present in the range of 0.01-99 wt%, more typically 30 to 80 wt%.
Other possible ingredients include antioxidants, colorants, flavors and fragrances, preservatives, saliva stimulating agents, cooling agents, co-solvents (including oils), lubricants, fillers, anti-foaming agents, surfactants, and taste masking agents.
The films of the present invention are typically prepared by evaporating and drying an aqueous film coated on a releasable backing or paper. This operation can be achieved in a drying oven or tunnel (typically a combination coater dryer), or by freeze drying or vacuum.
Solid preparations for oral administration may be prepared in immediate release and/or sustained release form. Sustained release formulations include delayed release, sustained release, pulsatile release, controlled release, targeted release and planned release.
Sustained release formulations suitable for the purposes of the present invention are described in U.S. Pat. No. 6,106,864. Other suitable drug delivery techniques (such as high energy dispersion and osmotic and coated granules) are described in detail in Verma et al, "Pharmaceutical technology on-line", 25(2), 1-14 (2001). Techniques for achieving controlled release using chewable tablets are described in WO 00/35298.
The compounds of the invention may also be administered directly into the bloodstream, muscle or internal organs. Suitable parenteral administration includes intravenous, arterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, and subcutaneous. Devices suitable for parenteral administration include needle (including microneedle) syringes, needleless syringes, and infusion techniques. Parenteral formulations are generally aqueous solutions which may contain excipients such as salts, carbohydrates and buffering agents, preferably at a pH of 3-9, although for some applications it may be more appropriate to formulate them as sterile nonaqueous solutions or in dry form for use with a suitable vehicle such as sterile pyrogen-free water. Preparation of parenteral formulations under sterile conditions (e.g., by lyophilization) can be readily accomplished using standard pharmaceutical techniques well known to those skilled in the art.
The solubility of the compounds of formula (I) for making parenteral solutions can be enhanced by using appropriate formulation techniques, such as the addition of solubility enhancers. Formulations for parenteral administration may be formulated for immediate release and/or sustained release. The sustained release preparation comprises delayed sustained release, sustained release, pulse sustained release, controlled release, targeted release and planned sustained release. Thus, the compounds of the present invention can be formulated as solids, semi-solids or thixotropic liquids, and used as an implanted depot delivery system that provides sustained release of the active compound. Examples of such formulations include drug-coated stents and suspensions of poly (dl-lactic-co-glycolic acid) (PGLA) microspheres.
The compounds of the invention may also be administered topically, transdermally (intra) or transdermally to the skin or mucosa. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, emulsions, ointments, dusting powders, dressings, foams, films, skin patches, drug-in-dry pastes, implants, sponges, fibers, bandages and microemulsions. Liposomes may also be used. Typical carriers include alcohols, water, mineral oil, liquid paraffin, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may be added, see for example J PharmSci, 88(10), 955-958 by Finnin and Morgan. Other topical administration includes by electroporation, iontophoresis, sonophoresis and microneedles or needle-free (e.g., Powderject)TM、BiojectTMEtc.) injection methods. Formulations for topical administration may be prepared as immediate release and/or sustained release. The sustained release preparation comprises delayed sustained release, pulse sustained release, controlled release, targeted release and planned sustained release.
The compounds of the invention may also be administered intranasally or by inhalation; administration is usually in the form of a dry powder from a dry powder inhaler, for example in the form of a dry mixture with lactose, alone or as admixed component particles, for example in admixture with a phospholipid, such as phosphatidylcholine. Or as an aerosol from a pressurized container, pump, spray, nebulizer (preferably one that uses electro-hydrodynamic force to produce a fine mist), or nebulizer, with or without the use of a suitable propellant, such as 1, 1, 1, 2-tetrafluoroethane or 1, 1, 1, 2, 3, 3, 3-heptafluoropropane, or as nasal drops. For intranasal administration, the powder may include a bioadhesive agent, for example, chitosan or cyclodextrin.
Pressurized containers, pumps, sprayers, atomizers or sprayers contain a solution or suspension of a compound of the invention, for example comprising ethanol, an aqueous solution of ethanol or a suitable alternative for dispersing, solubilizing or prolonging the release of the active compound, a propellant as a solvent and optionally a surfactant such as sorbitan trioleate, oleic acid or oligolactic acid.
Prior to use in dry powder or suspension formulations, the drug product is micronised to a size suitable for delivery by inhalation (typically below 5 microns). This can be accomplished by any suitable comminution method, such as spiral jet milling, fluidized bed jet milling, supercritical fluid processing to form nanoparticles, high pressure homogenization, or spray drying.
Capsules (made, for example, from gelatin or hydroxypropylmethyl cellulose), blisters and cartridges for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention, a suitable powder base such as lactose or starch, and a performance modifier such as 1-leucine, mannitol or magnesium stearate. Lactose can be anhydrous lactose or in the form of a monohydrate, the latter being preferred. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.
Solution formulations suitable for use in nebulizers that generate fine mist using electrohydrodynamics deliver a volume of 1 μ l to 100 μ l per 1 μ g to 20mg of the compound of the invention. Typical formulations may comprise a compound of formula (I), propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents that may be used in place of propylene glycol include glycerol and polyethylene glycol.
Suitable flavouring agents (such as menthol and levomenthol) or sweetening agents (such as saccharin or saccharin sodium) may be added to the formulations of the invention for inhalation/intranasal administration.
Formulations for inhalation/intranasal administration, for example, using PGLA, may be prepared as immediate release and/or sustained release. Sustained release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and planned release.
In the case of dry powder inhalers and aerosols, the dosage unit is determined by the valve delivering the metered dose. Dosage units according to the invention are generally designed to administer a metered dose or "shot size" containing from 1. mu.g to 100mg of a compound of formula (I). The total daily dose will typically range from 1 μ g to 200mg, which may be administered in a single dose, or more typically in multiple doses throughout the day.
The compounds of the invention may be administered rectally or vaginally, for example in the form of suppositories, pessaries, microbicides, pessaries or enemas. Cocoa butter is a traditional suppository base, but various alternatives may be used if appropriate. Formulations for rectal/vaginal administration may be prepared as immediate release and/or sustained release formulations. Sustained release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and planned release.
The compounds of the invention can also be administered directly to the eye or ear, typically in the form of droplets of a micronized suspension or solution in isotonic, pH-adjusted, sterile saline. Other formulations suitable for ocular and otic administration include ointments, biodegradable (e.g., absorbable gel sponges, collagen) and non-biodegradable (e.g., silicone) implants, drug-in-dry pastes, lenses, and particulate or vesicular systems, such as lipoidal vesicles or liposomes. The following polymers may be added along with preservatives such as benzalkonium chloride: such as crosslinked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, a cellulose polymer (e.g., hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose), or a heteropolysaccharide polymer (e.g., gellan gum). The formulation may also be delivered by iontophoresis. Formulations for ocular/otic administration may be prepared as immediate release and/or sustained release. Sustained release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release or planned release.
The compounds of the invention may be combined with soluble macromolecular entities such as cyclodextrins and suitable derivatives thereof or polyethylene glycol containing polymers for improved solubility, dissolution rate, taste masking, bioavailability and/or stability for any of the above modes of administration. It has been found that, for example, drug-cyclodextrin complexes are generally useful in most dosage forms and routes of administration. Inclusion complexes and non-inclusion complexes may be used. As an alternative to direct complexation (complexation) with the drug, cyclodextrins may be used as an auxiliary additive, i.e. as a carrier, diluent or solubiliser. The most commonly used for this purpose are alpha-cyclodextrin, beta-cyclodextrin and gamma-cyclodextrin, examples of which are found in international patent applications WO 91/11172, WO 94/02518 and WO 98/55148.
Since it may be desirable to administer a combination of active compounds, for example, in order to treat a particular disease or condition, the scope of the present invention encompasses the convenient combination of two or more pharmaceutical compositions, at least one of which contains a compound of the present invention, in a kit form suitable for co-administration. Thus, the kit of the invention comprises two or more separate pharmaceutical compositions, at least one of which contains a compound of formula (I) of the invention, and means for separately containing the compositions, such as a container, a divided bottle or a divided foil pouch. Examples of such kits are blister packs commonly used for packaging tablets, capsules and the like. The kit of the invention is particularly suitable for administration in different dosage forms, e.g. oral and parenteral administration, for administration of separate compositions at different dosing intervals, or for mutually titrating the respective compositions against each other. To aid compliance, the kit will typically contain instructions for administration and may provide so-called memory support.
For administration to a human patient, the total daily dose of a compound of the invention will generally range from 1mg to 10g, such as 10mg to 1g, such as 25mg to 500mg, depending, of course, on the mode of administration and the therapeutic effect. For example, oral administration may require a total daily dose of 50mg to 100 mg. The total daily dose may be administered in a single dose or in multiple doses and may be outside the general ranges specified herein at the discretion of the physician. These doses are based on an average human subject having a body weight of about 60kg to 70 kg. It is easy for a physician to determine dosages suitable for subjects with weights outside this range, such as young children and the elderly.
To improve dissolution characteristics, 4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] was prepared as follows]Pyridin-2-one (example 15, infra) with hydroxypropyl methylcellulose (HPMC E3 Prem LV, MethocelSolid amorphous spray-dried dispersion (SDD) available from Dow chemical company, Midland, MI). First, a spray solution containing 2.97g of water, 16.83g of methanol and 250. mu.L of 1M KOH (containing 9.8mg of potassium cation) was formed, and 51.27mg of 4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] was added thereto]Crystalline neutral form of pyridin-2-one. Then, 140.4mg HPMC was added to the solution and the solution was stirred for 5 minutes and sonicated for 2 minutes. The solution was pumped at a flow rate of 1.1ml/min using a Cole Parmer 74900 tandem speed control syringe pump into a small spray drying apparatus consisting of a stainless steel shell chamber with a diameter of 11 cm. Using a heated nitrogen stream, the solution was heated at 1 standard ft3The flow rate per minute was atomized with a two-fluid nozzle (Spraying Systems Co., Wheaton, Illinois, model: SU 1A). The hot gas enters the shell chamber at an inlet temperature of 85 ℃ and exits at an outlet temperature of 22 ℃. The resulting solid amorphous dispersion was collected on Whatman #1 microcrystalline cellulose filter media (11 μm pore size, 11cm outer diameter), dried under vacuum, and stored in a desiccator. The dispersion contained 25.4% by weight of 4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ]Pyridin-2-one, 4.9 wt% potassium cation and 69.7 wt% HPMC. The yield thereof was found to be about 60%.
For the avoidance of doubt, "treatment" as used herein includes reference to therapeutic, palliative and prophylactic treatment.
The following scheme illustrates a general process for the preparation of compounds of formula (I) and intermediates thereof.
It will be appreciated by those skilled in the art that some of the procedures described in the preparative schemes for the compounds of formula (I) or intermediates thereof may not be applicable to some of the possible alternative substituents.
It will also be appreciated by those skilled in the art that it may sometimes be necessary or preferable to carry out the transformations described in the schemes in a different order than that described, or to modify one or more of the transformations, to form the desired compound of formula (I).
It will furthermore be appreciated by those skilled in the art that, as illustrated in the schemes below, it may be necessary or preferable to protect one or more sensitive groups in the molecule at any stage in the synthesis of the compound of formula (I) in order to prevent unwanted side reactions. In particular, protection of amine groups may be required. The protecting groups used in the preparation of the compounds of formula (I) may be used in a conventional manner. See, for example, those described in "Protective Groups in organic Synthesis" 3 rd edition (John Wiley and Sons, 1999) of Theodora W Green and Peter G M Wuts, in particular Chapter 7, page 494 (for the Amino Group) ", which is incorporated herein by reference, and methods of removing such Groups.
Amino protecting groups boc, benzyloxycarbonyl, benzyl and acetyl are particularly suitable for the preparation of compounds of the formula (I) and intermediates thereof.
Unless otherwise indicated, R in the schemes1-R7And Y are as defined herein. PG (Picture experts group)1And PG2Is a nitrogen protecting group.
The compounds of formula (I) can be prepared as described in scheme 1, with schemes 1 being further illustrated below in preparative examples 1-27.
It will be appreciated that the amino protecting group N (PG1) (PG2) in formulas (XVII) to (XIX) may in some cases be N (H) PG1。
Scheme 1
Scheme 1 depicts various ways of preparing compounds of formula (I).
a) For example, a commercially available nitrile (II) or a nitrile (II) prepared by any of the standard methods described in the chemical literature is reacted with a haloacetate (III) (e.g., ethyl bromoacetate) in the presence of a Zn source. Tet. letters, 1997, 38, 443-446 describes that if basic conditions are applied to isolate the product, the conversion reaction can produce an enamine of the general structure (IV).
b) If acidic conditions are applied in step a) above, the ketoester (V) is produced, which can then be reconverted to the enamine (IV) in a separate step using an ammonia source, e.g.ammonium acetate. It follows that the various ketoesters of general structure (V) can intercept the above synthetic route at Intermediate (IV). c) Enamines (IV) may then be reacted with dialkyl malonates (VII) under basic conditions, such as sodium ethoxide, sodium hydride, or potassium tert-butoxide, to yield (VIII). Examples of such transformations are described in j. org. chem., 1981, 46(15), 3040-3048.
d) Alternatively, enamine (IV) may be reacted with malonyl chloride (VI) in the presence of a suitable base, such as potassium carbonate or triethylamine, to yield amidated form (IX).
e) (IX) can then be reacted with a suitable base in a separation step to cyclize to pyridine (VIII). Suitable bases include sodium ethoxide, sodium hydride or potassium tert-butoxide.
f) (VIII) is then saponified under acidic or basic conditions with heating to provide the corresponding acid (e.g., HCl, HBr, sulfuric acid), sodium hydroxide or lithium hydroxide, and is then spontaneously decarboxylated to produce pyridine (X). An example of this conversion is described in WO 01101949.
g) (X) nitration to produce nitropyridine (XI) can be carried out using any literature conditions known to those skilled in the art, for example using a mixture of nitric acid and sulfuric acid or a mixture of acetic acid and nitric acid. This transformation is described, for example, in bioorg.med.chem.lett., 1996, 6(2), 173-.
h) (XI) can be chlorinated to give (XII) using various conditions for conversion of the hydroxyl group to chlorine, such as thionyl chloride or phosphorus oxychloride. It will be appreciated by those skilled in the art that 2 hydroxyl groups may be respectively chlorinated or converted to alternative leaving groups (such as another halogen atom) or activated esters (such as methane sulfonate or trifluoromethyl sulfonate). Examples of all such methods are included in the following sections.
i) Reacting (XII) with a compound of the formula R3YNH2To give (XIII). Vision R3Depending on the nature of the Y group, there may be partial metathesis of 2 chloro groups or a small amount of metathesis at the 2-chloro group without compromising the ability to significantly obtain (XIII).
j) (XIII) is then reacted with ammonia or an ammonia equivalent, such as ammonium acetate, to produce (XIV).
k) (XIV) can then be reduced under any of the conditions known in the literature (e.g., using iron or tin in HCl) to reduce the nitroaromatic compound to an amine, and then subjected to a hydrogenation reaction in the presence of a transition metal catalyst (such as palladium, platinum, or nickel) or a chemical reducing agent (such as lithium aluminum hydride) to produce (XV).
l) can then be reacted with a C ═ 0 source, such as 1, 1-carbonyldiimidazole or phosgene, to produce a mixture of imidazolone (XVI) and (I), which can be obtained by careful chromatographic purification.
m) or (XIII) can be reacted with a protected form of ammonia (such as dibenzylamine or diallylamine) in which 2 hydrogen atoms are substituted with 2 groups that are subsequently removed under mild conditions. For alternatives to these examples, see, for example, those Groups described in "Protective Groups in Organic Synthesis" 3 rd edition (John Wileyand Sons, 1999), in particular Chapter 7, page 494-653 ("Protection for the amino Group") of Theodora W Green and Peter G M Wuts.
n) -o) see steps k) -l).
p) then removing the protecting group PG where appropriate1And PG2Under various conditions to yield (I). See, for example, those described in "Protective Groups in Organic Synthesis" 3 rd edition (John Wiley and sons, 1999), in particular Chapter 7, page 494-653 ("Protection for the amino group"), of Theodora W Green and Peter G M Wuts, and the conditions for their removal.
q) alternatively, (XIII) it may be preferred to use as alkyl carbamate R a combination of a strong base such as sodium hydride, potassium tert-butoxide or lithium diisopropylamineamine with a suitable acylating agent such as ethyl chloroformate or any other alkyl or aryl chloroformate, chloroform or anhydride13CO2X (wherein R13Is C1-6Alkyl and X is halogen) to form (XX).
r) Steps j) and k) can be applied as above to (XX), and the product resulting from this sequence of steps can then be regioselectively cyclized to (I) by simple treatment with a protic acid, such as acetic acid or formic acid.
The compounds of general formula (I) can also be prepared according to scheme 2 below using an alternative configuration of intermediate (XVII) in which a halogen atom is present at C2 and a suitable intermediate (XXVII in scheme 2) is formed for further manipulation.
Scheme 2
Thus, commercially available dihalopyridines (e.g., dibromopyridine XXI) can be treated in several steps to yield analogs of formula (I).
a) -e) commercially available dibromopyridine (XXI) can be manipulated according to modified literature procedures (described in WO 2005026164) to yield intermediate (XXVI). The steps between (XXI) and (XXVI) depend on thermally dangerous agents or the production of potentially thermally dangerous products and should therefore be handled with caution.
f) (XXVI) is obtained by reaction using various conditions known to generate the diazonium species from an amino group, for example, nitrous acid conditions generated in situ from HCl and sodium nitrite wherein the diazonium species is substituted with a C1 atom. The conversion of (XXVI) to the key intermediate (XXVII) can then be achieved by adding an excess of amine (e.g. benzylamine) to the crude chloride. This step replaces the C2 halogen atom with the C4 halogen atom to an amino group. Any primary or secondary amine group is suitable for this conversion.
g) I) the C2 halogen atom in (XXVII) can then be reacted under conditions in which various substituents C2 halogen groups are present as various functional groups, whereby a variety of alternative products (I) can be prepared. For example, coupling of a heterocycle to (XXVII) using various organometallic reagents (boronic acid, zincate, magnesium reagent, cuprate, stannane, etc.) can yield (XXVIII) using vinyl organometallic reagents (such as vinyl tributyltin) and palladium catalysts (such as Pd (PPh) 3)4) Coupling of the heterocycle with (XXVII) produces the vinyl species (XXX), and a carbonylation reaction in which (XXVII) is treated with CO gas under pressure in the presence of a base such as triethylamine and a palladium catalyst to produce the acyl product of general structure (XXIX). Compounds of structures (XXVIII) and (XXIX) can then be converted to compounds of formula (I) according to steps n, o and p of scheme 1.
j) -l) (XXX) can then be treated by oxidation to give the aldehyde (XXXII), then either (XXXII) or (XXXI) can be treated with an amine in the presence of a base such as triethylamine or with a reducing agent such as sodium triacetoxyborohydride, respectively, to give the product (I) (following steps n, o and p in scheme 1).
Alternative configurations of intermediate (XIII) can be prepared according to scheme 3, and then operated as follows to yield the compound of formula (I).
Scheme 3
a) -e) steps a) -e) are essentially similar to those described in scheme 1, except that a monohalide (XXXIV) is prepared to ensure positional chemical integrity of C6 and C4 in (XXXVI).
f) (XXXVIII) can be brominated under various conditions known to those skilled in the art, such as bromine water in a non-polar solvent such as DCM or MeCN, to yield C3-bromide (XXXIX).
g) CO gas may then be used in the presence of a base such as triethylamine, an alcohol such as methanol, and a base such as Pd (PPh)3)4Carbonylating (XXXIX) in the presence of an isopalladium catalyst under pressure to produce an ester (XXXX).
h) I) then (XXXX) can be reacted with an amine NR6R7Reaction to give amide (XXXXI), which can be deprotected as above to give product (I).
An alternative synthesis of intermediate (XI) is shown in scheme 4 below.
Scheme 4
a) Any nitrile (XXXXII) having a methylene group adjacent to the nitrile function can be reacted with malonyl chloride (xxxiii) to form pyridine (xxxxxiv). This transformation is described in Synthesis, 1984, 765 and 766.
b) Pyridine (XXXXIV) may then be nitrated to form nitropyridine (XXXXV) using any literature conditions known to those skilled in the art, such as using a mixture of nitric acid and sulfuric acid or a mixture of acetic acid and nitric acid.
c) The chlorine atom in (XXXXV) may then be coupled under various conditions, which may be in the presence of a transition metal catalyst (in the presence of a palladium catalyst)Below) with a reactive organometallic reagent (e.g., stannane, zincate, or boronic acid) to yield a substituted pyridine (XI). In which R is1When H, a simple hydrogenation reaction can be effectively carried out in the presence of a suitable catalyst such as palladium on charcoal or palladium hydroxide.
Another alternative synthesis of intermediate (X) is shown in scheme 5 below.
Scheme 5
In this process, a malonyl ester, preferably diphenyl ester (xxxxxvii), is reacted with a schiff base (xxxxxvii) and heated to produce a pyridine of general formula (X).
Another alternative way of preparing the compound of general formula (X) is shown in scheme 6.
Scheme 6
a) Ketoacids of general formula (xxxxxii), which are commercially available or directly prepared by saponification of a ketone ester of formula (V), are reacted with a C ═ 0 source, such as 1, 1-carbonyldiimidazole, in a suitable solvent at elevated temperature to produce the cyclic pyrones (XXXXIX).
b) (XXXXXXIX) is treated with a strong mineral acid such as sulphuric acid or hydrochloric acid to remove the C3-acyl substituent and give pyrone (L).
c) (L) can be reacted with an ammonia source such as concentrated aqueous ammonium hydroxide under heating to convert the pyrone ring to pyridine (X), thereby capturing the same intermediates described in scheme 1. Alternatively, several pyrones of general formula (L) are available from commercial sources, which can be directly applied in step c. The conversion of compounds of formula (L) to compounds of formula (X) is described in several sources, for example WO 9504730.
An alternative way of preparing the compound of general formula (XIX) is shown in scheme 7.
Scheme 7
a) Commercially available dichloropyridines of the general formula (LI) can be reacted with compounds of the formula R3YNH2To selectively substitute the 4-chloro group, thereby producing pyridine (LII).
b) (LII) can then be reacted with a protected form of ammonia PG1NH2Or PG1PG2NH (such as benzylamine, allylamine, dibenzylamine, or diallylamine) reacts to replace the 2-chloro group, where 2 hydrogen atoms are replaced with 2 groups that can then be removed under mild conditions. For alternatives to this example, see for example those described in "protective groups in Organic Synthesis" 3 rd edition (John Wiley and Sons, 1999), in particular Chapter 7, page 494-653 ("Protection for the Amino Group"), of the Theodora W Greene and Peter G M Wuts. If an excess of amine R is used from step a3YNH2This group may then replace the 2-chloro group with the 4-chloro group.
c) The ester of (LIII) can be hydrolyzed under various conditions known to deprotect the ester (e.g., sodium hydroxide or lithium hydroxide) to produce an acid (LIV). For alternatives to this agent, see for example those conditions described in "Protective Groups in organic Synthesis" 3 rd edition (John Wiley and Sons, 1999), in particular Chapter 5, page 373-441 ("Protection for the Carboxylgroup") of the Theodora W Greene and Peter G M Wuts.
d) The (LIV) can then be reacted with reagents known to convert acids to acyl azides (e.g., diphenylphosphonyl azide). Upon heating, the intermediate acyl azide undergoes a rearrangement reaction in which isocyanate is produced and trapped internally by a 4-amino substituent to produce imidazolone (XIX), thereby trapping the same intermediate described in scheme 1.
An alternative way of preparing the compounds of general formula (I) is shown in scheme 8.
Scheme 8
a) (LVI) prepared according to org.BioMol.chem., 2003, 1, 1354-1365 can be halogenated by reacting the amino group in (LVI) with an active agent (e.g., isoamyl nitrite) and a halogenating agent (such as diiodomethane or dibromomethane) to produce a halogenated species (LVII).
b) (LVII) may then undergo various transition metal mediated coupling reactions in which the iodine group selectively reacts with, for example, a terminal alkyne. See j.med.chem., 34(2), 1991, 778-786 for an example of this transformation to the imidazole configuration.
c) (LVIII) can then be reacted with ammonia to cyclize the alkynyl-carbonitrile to a pyridine ring. See, for example, Tetrahedron, 49(3), 1993, 557-570 for examples of such transformations on the imidazole configuration.
d) The bromine atom in (LIX) can then be hydrolyzed under strongly acidic conditions (e.g., hydrochloric acid or sulfuric acid); or with a nucleophilic source of OH, such as sodium hydroxide or sodium methoxide, followed by mild acidic hydrolysis to yield (I).
Intermediate (LVIII) can also be used to prepare compounds of general formula (I) according to scheme 9 below.
Scheme 9
e) The (LVIII) can be reacted with an alcohol (e.g., methanol, ethanol, propanol, or any other alcohol) under mild heating to form the acetal (LX).
f) The acetal (LX) can then undergo hydrolysis under any conditions known to hydrolyze acetals or ketals to ketones (e.g., aqueous hydrochloric acid). For alternatives to this condition, see for example those conditions described in "Protective Groups in organic Synthesis" 3 rd edition (John Wiley and Sons, 1999), in particular Chapter 4, page 297-.
g) The ketone (LXI) can then be alkylated in the presence of a suitable base such as sodium hydride, potassium tert-butoxide or potassium carbonate, and an alkylating agent such as an alkyl halide, alkyl sulfonate or alkyl triflate to produce (LXII).
h) The (LXII) can then be reacted with ammonia to cyclize the ketone-carbonitrile to a pyridine ring.
i) The bromine atom in (LXIII) can then be hydrolyzed under strongly acidic conditions (e.g., hydrochloric acid or sulfuric acid); or with a nucleophilic source of OH, such as sodium hydroxide or sodium methoxide, followed by mild acidic hydrolysis to yield (I). During the alcoholysis step e, the high temperature can cause the bromine atom to be substituted by an alcohol, which is then introduced into (I) in an alternative way to the introduction of an oxo group.
Schemes 8 and 9 are illustrated using bromine, which is preferred, but it is understood that other halogen atoms can also be used.
An alternative synthesis of intermediate (XVII) is shown in scheme 10 below.
Scheme 10
a) (LXIV) can be treated under any nitration conditions known to those skilled in the art. This type of transformation is known to be performed by using an intermediate N-nitro analogue. See, for example, the same chemistry described in WO 2005026164.
b) (LXV) is treated with HBr to convert the chlorine atom to bromine (LXVI).
c) (LXVI) is then treated with any conditions known to those skilled in the art to convert an amino group to an N-nitroso or diazo group, followed by treatment with HCl to yield chloride (LXVII).
d) (LXVII) useful amine PG 1NH (e.g., benzylamine) to replace 4-chloro and 2-bromo to yield (LXVIII).
e) The remaining 6-bromine can then be used to introduce various substituents using transition metal mediated methods, e.g., Pd catalyzed carbonylation, organometallic cross-coupling reactions via Sn, Zn, or B reagents, or using Li or Mg reagents using Fe or Ni as catalysts. In the examples shown, the carbonylation reaction produced the specific analog shown (LXIX). This can then be converted to the compound of formula (I) according to steps n, o and p in scheme 1.
An alternative way of preparing the compound of formula (XIX) is shown in scheme 11.
Scheme 11
a) (LXX) is halogenated in turn under various conditions that can introduce halogen atoms (e.g., NCS, NBS, NIS, bromine water, etc.) to produce the dihalopyridine (LXXI).
b) Then using the general structure R3YNH2To yield a mixture of two compounds (LXXII) and (LXXIII).
c) -e) the desired compound (LXXII) can then be cyclized, replacing the remaining halogen, and then finally deprotected, to give (I).
Another method for preparing compounds of formula (X), (XI) and (I) is shown in scheme 12.
Scheme 12
a) Thus, a general dehydrating agent (such as POCl) can be used3、SOCl2PPA or triflic anhydride), by reacting a commercially available amide of formula (LXXV) with a malonyl diester (e.g., dialkyl 1, 3-acetonedicarboxylate) via dehydration of the starting amide in the presence of a strong base under conditions such that a nitrile can be formed in situ. This condition directly leads to the formation of the dihydroxypyridine (X).
b) (X) can be nitrated according to the method described in scheme 1 to yield (XI).
c) (X) saponification and decarboxylation can be performed according to the method described in scheme 1.
d) And e) either (X) or (XI) can then be converted to the compound of formula (I) using any of the methods described in scheme 1.
(I) The preparation of prodrug derivatives of (a) is shown in scheme 13 below.
Scheme 13
Active parent compound and characterized by the group R4Or R5Reaction of a reagent attached to a suitable leaving group in the presence of a suitable base may result in a prodrug derivative of (I). Suitable reagents include, but are not limited to, alkyl halides, acid chlorides, chloroformates, and carbamoyl chlorides as shown below.
Suitable bases include triethylamine, diisopropylethylamine, potassium carbonate, cesium carbonate, sodium hydride and n-butyllithium. Various solvents (including but not limited to THF, acetonitrile, dimethylformamide, dichloromethane, and diethyl ether) may also be used to effect this conversion. The specific choice of solvent and base will influence the regioselectivity of the alkylation/acylation reaction, i.e., whether the reactive group is attached to an O atom (R) 5) Or N atom (R)4). For example, the reaction of the parent molecule with ethyl chloroformate in the presence of triethylamine in DCM can significantly produce O acylation.
The preparation of all the reactions described above as well as the novel starting materials disclosed in the above methods is conventional and suitable reagents and reaction conditions for their performance or preparation as well as procedures for isolating the desired products are known to those skilled in the art from reference precedent and examples and preparations thereof.
The compounds of the present invention are useful for their pharmacological activity in mammals, including humans. In particular, it is useful for the treatment of diseases associated with the modulation (especially agonism) of TLR 7.
In another embodiment, the invention additionally provides a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a disease or disorder associated with the modulation of the TLR7 receptor.
The present invention thus provides a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a disease or condition in which modulation of the TLR7 receptor is known or can be demonstrated to produce a beneficial effect.
In yet another embodiment, the compounds of the invention are useful for treating viral infections, such as those caused by: adenovirus, herpesvirus (e.g., HSV-I, HSV-II, CMV, or VZV), poxvirus (e.g., orthopoxvirus such as smallpox or vaccinia, or molluscum contagiosum), picornavirus (e.g., rhinovirus or enterovirus), orthomyxovirus (e.g., influenza virus), paramyxovirus (e.g., parainfluenza virus, mumps virus, measles virus, or Respiratory Syncytial Virus (RSV)), coronaviruses (e.g., SARS), papovaviruses (e.g., papilloma viruses, such as those producing genital warts, common warts, or plantar warts), hepadnaviruses (e.g., hepatitis b virus), flaviviruses (e.g., hepatitis c virus or dengue virus), retroviruses (e.g., lentiviruses such as HIV), or filoviruses (e.g., ebola virus or marburg virus).
In another embodiment, the compounds of the invention are useful for treating hepatitis c virus infection.
In yet another embodiment, the compounds of the invention are useful for the treatment of tumors or cancers, including but not limited to carcinomas, sarcomas, and leukemias, e.g., squamous cell carcinoma, renal cell carcinoma, kaposi's sarcoma, melanoma, renal cell carcinoma, myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, non-hodgkin's lymphoma.
In yet another embodiment, the compounds of the present invention are useful for the treatment of bacterial, fungal and protozoal infections, including but not limited to infections caused by bacteria of the genera: escherichia (Escherichia), Enterobacter (Enterobacter), Salmonella (Salmonella), Staphylococcus (Staphylococcus), Klebsiella (Klebsiella), Proteus (Proteus), Pseudomonas (Pseudomonas), Streptococcus (Streptococcus), Chlamydia (Chlamydia); or fungal infections such as candidiasis (candidiasis), aspergillosis (aspergillosis), histoplasmosis (histoplasmosis), cryptococcal meningitis (cryptococcal meningitis).
In yet another embodiment, the compounds of the invention are useful in the treatment of T-helper (Th2) mediated diseases (see, e.g., Dabbagh et al, Curr Opin infection Dis 2003, 16: 199-204, incorporated herein by reference), including but not limited to, atopic diseases such as atopic dermatitis or eczema, hypereosinophilia, asthma, allergy, allergic rhinitis.
In yet another embodiment, the compounds of the present invention are useful for treating damaged or aged skin, such as scars and wrinkles.
In yet another embodiment, the compounds of the invention are useful in the treatment of autoimmune diseases, such as crohn's disease and inflammatory bowel disease.
The compounds of formula (I) and pharmaceutically acceptable salts or solvates thereof may be administered alone or as part of a combination therapy. Thus, the scope of the present invention includes examples of compositions comprising a compound of the present invention and one or more additional therapeutic agents, as well as examples of co-administration thereof.
In one aspect, the combination forms of the invention include treatment with a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and one or more other agents having anti-HCV activity, which are agents that inhibit the following targets, such as, but not limited to: HCV NS3 protein, HCV NS5A protein, HCV NS4B protein, HCV polymerase, HCV metalloprotease, HCV serine protease, HCV helicase, p7 protein. Examples of such agents include, but are not limited to: interferons, pegylated interferons (e.g., pegylated alpha-2 a and pegylated alpha-2 b interferons), long-acting interferons (e.g., albumin interferon alpha), lamivudine (lamivudine), ribavirin (ribavarin), emtricitabine (emtricitabine), vilaminidine (viramidine), cigivir (celgosivir), valopicitabine (valacitabine), HCV-086, HCV-796, RMZ702, BILN2061, IDN6566, NM283, SCH 6, and VX-950.
In another embodiment, the combination of the invention comprises treatment with a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and one or more other TLR agonists such as an agonist of the TLR3, TLR7, TLR8 or TLR9 receptor.
In another embodiment, the combination of the invention comprises the treatment of HCV-HIV co-infection with a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and one or more other antiviral agents selected from the following inhibitors: HIV Protease Inhibitors (PI), non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleoside/Nucleotide Reverse Transcriptase Inhibitors (NRTIs), CCR5 antagonists, agents that inhibit the interaction of gp120 with CD4, agents that inhibit HIV entry into target cells (e.g., fusion inhibitors), integrase inhibitors, prenylation inhibitors, RNase H inhibitors, and maturation inhibitors.
Examples of NNRTIs include, but are not limited to: efavirenz (efavirenz), HBY-097, nevirapine (nevirapine), TMC-120 (dapivirine), TMC-125, etravirine (etravirine), delavirdine (delavirdine), DPC-083, DPC-961, cappivirine (capravirine), rilpivirine, 5- { [3, 5 diethyl-1- (2-hydroxyethyl) -1H-pyrazol-4-yl ] oxy } isophthalonitrile, or a pharmaceutically acceptable salt, solvate, or derivative thereof; GW-678248, GW-695634, MIV-150, calanolide (calanolide), and tricyclic pyrimidinone derivatives, as disclosed in WO 03/062238.
Examples of CCR5 antagonists include, but are not limited to: TAK-779, SC-351125, Anlivir Rou (anciriroc) (formerly SCH-C), Virivir Rou (viciriroc) (formerly SCH-D), maravir Rou (maraviroc), PRO-140, Alraviro (apliviroc) (formerly GW-873140, Ono-4128, AK-602), AMD-887 CMPD-167, 1-endo- {8- [ (3S) -3- (acetylamino) -3- (3-fluorophenyl) propyl ] -8-azabicyclo [3.2.1] oct-3-yl } -2-methyl-4, 5, 6, 7-tetrahydro-1H-imidazo [4, 5-C ] pyridine-5-carboxylic acid methyl ester or a pharmaceutically acceptable salt, solvate or derivative thereof, 3-endo- {8- [ (3S) -3- (acetylamino) -3- (3-fluorophenyl) propyl ] -8-azabicyclo [3.2.1] oct-3-yl } -2-methyl-4, 5, 6, 7-tetrahydro-3H-imidazo [4, 5-c ] pyridine-5-carboxylic acid methyl ester or a pharmaceutically acceptable salt, solvate or derivative thereof, 1-endo- {8- [ (3S) -3- (acetylamino) -3- (3-fluorophenyl) propyl ] -8-azabicyclo [3.2.1] oct-3-yl } -2-methyl-4, 5, 6, 7-tetrahydro-1H-imidazo [4, 5-c ] pyridine-5-carboxylic acid ethyl ester or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable acid, a pharmaceutically acceptable salt thereof, and a, Solvates or derivatives, and N- { (1S) -3- [ 3-endo- (5-isobutyryl-2-methyl-4, 5, 6, 7-tetrahydro-1H-imidazo [4, 5-c ] pyridin-1-yl) -8-azabicyclo [3.2.1] oct-8-yl ] -1- (3-fluorophenyl) propyl } acetamide, or a pharmaceutically acceptable salt, solvate, or derivative thereof.
Examples of entry and fusion inhibitors include, but are not limited to: BMS-806, BMS-488043, 5- { (1S) -2- [ (2R) -4-benzoyl-2-methyl-piperazin-1-yl ] -1-methyl-2-oxo-ethoxy } -4-methoxy-pyridine-2-carboxylic acid methylamide and 4- { (1S) -2- [ (2R) -4-benzoyl-2-methyl-piperazin-1-yl ] -1-methyl-2-oxo-ethoxy } -3-methoxy-N-methyl-benzamide, enfuvirtide (T-20), Sifuvirtide SP-01A (sifuvirtide SP-01A), T1249, PRO 542, AMD-3100, soluble CD4, a compound disclosed in JP 2003171381 and a compound disclosed in JP 2003119137.
Examples of HIV integrase inhibitors include, but are not limited to: l000870810, GW-810781, 1, 5-naphthyridine-3-carboxamide derivatives disclosed in WO 03/062204, compounds disclosed in WO 03/047564, compounds disclosed in WO 03/049690 and 5-hydroxypyrimidine-4-carboxamide derivatives disclosed in WO 03/035076, MK-0518 (5- (1, 1-dioxo-1, 2-thiadiazoheterocyclohexan-yl (thiazinan) -2-yl) -N- (4-fluorobenzyl) -8-hydroxy-1, 6-naphthyridine-7-carboxamide disclosed in WO 03016315), GS-9137 (JTK-303).
Examples of prenylation inhibitors include, but are not limited to: HMG coa reductase inhibitors such as statins (e.g., atorvastatin).
Examples of maturation inhibitors include 3-O- (3 '3' -dimethylsuccinyl) betulinic acid (otherwise known as PA-457) and alpha HGA.
In yet another aspect, the combination forms of the invention include treatment with a compound of formula (I) or a pharmaceutically acceptable salt, solvate or polymorph thereof and one or more additional agents such as, but not limited to: antifungal agents, such as fluconazole (fluconazole), fosfluconazole (fosfluconazole), itraconazole (itraconazole), or voriconazole (voriconazole); antibacterial agents, for example, azithromycin (azithromycin) or clarithromycin (clarithromycin); interferons, daunorubicin (daunorubicin), doxorubicin (doxorubicin), and paclitaxel (paclitaxel) for treatment of AIDS-related kaposi's sarcoma; and cidofovir (cidofovir), fomivirsen (fomivirsen), foscarnet (foscarnet), ganciclovir (ganciclovir) and valacyclovir (valcyte) for the treatment of Cytomegalovirus (CMV) retinitis.
In yet another embodiment, the combination of the invention comprises treatment with a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and one or more other therapeutic agents that enhance the immune system of the body, including low doses of cyclophosphamide, thymic stimulin, vitamins and nutritional supplements (e.g., antioxidants including vitamin A, C, E, beta-carotene, zinc, selenium, glutathione, coenzyme Q-10, and echinacea), and vaccines such as Immune Stimulating Complexes (ISCOMs) comprising a vaccine formulation combining an antigen expressing multimer 5 with an adjuvant.
Other combinations for use according to the invention include a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof in combination with an agent which is: CCR1 antagonists such as BX-471; beta adrenergic receptor agonists such as salmeterol (salmeterol); corticosteroid agonists, such as fluticasone propionate; LTD4 antagonists, such as montelukast (montelukast); muscarinic antagonists such as tiotropium bromide (tiotropium bromide); PDE4 inhibitors, such as cilomilast (cilomilast) or roflumilast (roflumilast); COX-2 inhibitors such as celecoxib (celecoxib), valdecoxib (valdecoxib), or rofecoxib (rofecoxib); alpha-2-delta ligands such as gabapentin (gabapentin) or pregabalin (pregabalin); TNF-alpha receptor modulators, such as TNF-alpha inhibitors (e.g., adalimumab); or immunosuppressive agents such as cyclosporine or macrolides such as tacrolimus (tacrolimus).
Also included within the scope of the present invention are combinations of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and one or more other therapeutic agents which reduce the rate of metabolism of the compound of the present invention, thereby causing increased contact with the patient. Increasing exposure in this manner is known as boosting. This effect has the advantage of either enhancing the efficacy of the compounds of the invention or reducing the dosage required to achieve the same efficacy as the non-synergistic dosage. The metabolism of the compounds of the invention includes oxidation by P450(CYP450) enzymes, especially CYP3a4, and the combination by UDP glucuronidase and sulfurylase. Thus, among the agents that may be used to increase patient contact with the compounds of the present invention are those that are capable of acting as inhibitors of at least one isoform of cytochrome P450(CYP450) enzyme. Isoforms of CYP450 that may be advantageously inhibited include, but are not limited to: CYP1a2, CYP2D6, CYP2C9, CYP2C19 and CYP3a 4. Suitable agents that may be used to inhibit CYP3a4 include, but are not limited to: ritonavir (ritonavir), saquinavir (saquinavir), or ketoconazole (ketoconazole).
In the above combinations, the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and the other therapeutic agent may be administered alone or in combination with each other in terms of dosage form; the administration may be simultaneous or sequential in terms of time of administration. Thus, administration of one component agent may be prior to, simultaneous with, or subsequent to administration of the other component agent.
It is to be understood that reference herein to treatment includes therapeutic, palliative and prophylactic treatment.
It should be understood that the present invention includes the following aspects.
(i) A compound of formula (I) or a tautomer thereof or a pharmaceutically acceptable salt or solvate of the compound or tautomer;
(ii) a pharmaceutical composition comprising a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of said compound or tautomer, as claimed in any one of the preceding inventions, and one or more pharmaceutically acceptable excipients;
(iii) a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of said compound or tautomer, for use as a medicament;
(iv) a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or tautomer, for use in the treatment of a disease or disorder associated with the modulation of the TLR7 receptor;
(v) Use of a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or tautomer, in the manufacture of a medicament for the treatment of a disease or disorder associated with the modulation of TLR7 receptors.
(vi) Pharmaceutical compositions comprising one or more other therapeutic agents;
(vii) a pharmaceutical product (e.g. in the form of a kit) comprising a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of said compound or tautomer, and a further therapeutically active agent, as a combined preparation for simultaneous, separate or sequential use in the treatment of a disease associated with the modulation of the TLR7 receptor.
(viii) Use of a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of said compound or tautomer, in the manufacture of a medicament for simultaneous, separate or sequential use in combination with other therapeutically active agents in the treatment of a disease associated with the modulation of the TLR7 receptor.
(ix) A method of treating a disease or disorder associated with modulation of TLR7 receptors in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or tautomer.
(x) A process for preparing a compound of formula (I) or a tautomer thereof, or a pharmaceutically acceptable salt or solvate of said compound or tautomer.
(xi) Some novel intermediates disclosed herein.
Detailed Description
The invention is illustrated by the following non-limiting examples, in which the following abbreviations and definitions are used:
filter (lignocellulose) from J.
Rettenmaier & Sohne，Germany
APCI+Atmospheric pressure chemical ionization (forward scan)
Bn benzyl group
br broad peak
d double peak
dd doublet of doublets
DMSO dimethyl sulfoxide
ELSD evaporative light scattering detector
ES+Electrospray forward scanning mass spectrometry
ESI electrospray ionization mass spectrometry
(Positive or negative going scan)
eq equivalent weight
HRMS high resolution mass spectrometry
1H NMR proton nuclear magnetic resonance
LC-MS liquid chromatography-mass spectrometer
LRMS low resolution mass spectrometry
m multiplet
Peak of mass m/z
Reacti-VialTMReaction flask, from USA
Feishale scientific world Co Ltd
q quartet peak
s single peak
t triplet peak
Chemical shift of delta
Indicates the connection point
Example 1
4-amino-1-benzyl-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
4-allylamino-1-benzyl-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (70mg, 0.2mmol) was dissolved in ethanol (2ml), 10% Pd-C (70mg, w/w) was added, and BF was added dropwise3·OEt2(271, 0.2 mmol). Mixing the mixture in N2Under the atmosphere,Heat at reflux overnight. The mixture was cooled to room temperature and filtered through Arbocel (lignocellulose), rinsed with fresh EtOH and the filtrate was concentrated in vacuo to give the crude product (150 mg). Elution with 98:2 → 95:5 DCM: MeOH followed by silica column chromatography gave the title compound as an off-white solid (17 mg).
1H NMR(CD3OD)δ 7.35-7.27(m，5H)，6.26(s，1H)，5.00(s，2H)，1.89-1.82(m，1H)，0.85-0.80(m，2H)，0.77-0.73(m，2H)；C16H16N4HRMS of O: calculated 281.1397, found 281.1395.
Example 2
4-amino-1-benzyl-6-methyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-dibenzylamino-6-methyl-1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (34mg, 0.08mmol) was suspended in ethanol (5ml) and incubated at room temperature at 60psi in 10% Pd (OH)2(7mg) was hydrogenated for 6 hours. The reaction mixture was filtered through a short plug of Arbocel and the filtrate was evaporated in vacuo to an opaque gum. This gel was dissolved in methanol and pre-adsorbed on silica gel, then purified by column chromatography eluting with 5% methanol in EtOAc. The appropriate fractions were combined and evaporated in vacuo to give the title compound as a white solid, 7 mg.
1H NMR(CD3OD)δ 2.31(s，3H)，5.01(s，2H)，6.40(s，1H)，7.31(m，5H)；LRMS(ES+)m/z 255(MH+)。
Example 3
1-benzyl-4-amino-6, 7-dimethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-diallylamino-6, 7-dimethyl-1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (358mg, 1mmol) was dissolved in water (10ml) and acetonitrile (25ml), and RhCl (PPh) was added in one portion3)3(286mg, 0.3mmol) and the mixture was then heated at reflux for 16 h. The mixture was cooled to room temperature and then concentrated in vacuo, and the residue was purified by silica gel column chromatography using a gradient of 95:5 → 85:15 DCM: MeOH to give the title compound as a light brown solid (77mg, 29%).
1H NMR(DMSO)δ 1.98(s，3H)，2.14(s，3H)，5.13(s，2H)，5.34(s，2H)，7.03-7.31(m，5H)，10.38(s，1H)；LRMS(ES+)m/z 269[MH]+
Example 4
4-amino-1-benzyl-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylic acid methyl ester
Methyl 1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylate (0.02g) was dissolved in sulfuric acid (2ml) and stirred rapidly for 15 minutes. The reaction mixture was cooled to 0 ℃, water was added to give a precipitate, which was filtered and dried in vacuo to give the title compound as a white solid (10 mg).
1H NMR(d6-DMSO，400MHz)δ 3.75(s，3H)，4.95(s，2H)，7.20-7.50(m，6H)，10.20(s，1H)。LRMS(ES+)m/z 299[MH]+
Example 5
4-amino-1-benzyl-6-pyrazin-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
N-2, N-4-dibenzyl-6-pyrazin-2-yl-pyridine-2, 3, 4-triamine (100mg/0.261mmol) was dissolved in DMF (5ml), CDI (95.3mg/0.523mmol) was added, heated at 60 ℃ for 5 hours, and then concentrated in vacuo. The reaction mixture was then dissolved in concentrated sulfuric acid (3ml) and stirred at room temperature for 30 minutes. Ice was then added to the reaction, by pouring K in water (5ml)2CO3(8g) To quench. It was then extracted with EtOAc, dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography. In detail, the two regioisomers were then separated using EtOAc, 95:5EtOAc: MeOH to give the title compound as a light orange solid (15 mg).
1H NMR(CD30D，400MHz)δ 5.10(s，2H)，7.20-7.40(m，5H)，7.55(s，1H)，8.45(d，1H)，8.55(d，1H)，9.4(s，1H)
Example 6
4-amino-1-benzyl-6-morpholin-4-ylmethyl-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one
N-2, N-4-dibenzyl-6-morpholin-4-yl-methyl-pyridine-2, 3, 4-triamine (240mg, 0.59mmol) was dissolved in 20ml dichloromethane, then 1, 1' -carbonyldiimidazole (91mg, 1.77mmol) was added and the reaction was stirred at room temperature for 48 hours. 20ml of water was added and the organic layer was separated, dried over magnesium sulfate and the solvent removed in vacuo. The crude residue was purified by silica gel column chromatography using 4% methanol in dichloromethane to yield 140mg of a mixture of 2 isomers. 60mg of this mixture of 2 isomers are dissolved in 2ml of concentrated sulfuric acid and stirred at room temperature for 30 minutes. Water (5ml) was added cautiously followed by potassium carbonate (5.2g until pH 7). The mixture was extracted with ethyl acetate, the organic layer was separated, dried over magnesium sulfate and the solvent was removed in vacuo. The crude residue was purified by silica gel column chromatography using 1% ammonia and 10% methanol in dichloromethane to give 10mg of the title compound and 6mg of the other isomer.
1H NMR(CD3OD)：δ 7.38-7.2(m，5H)，6.58(s，1H)，5.05(d，2H)，3.65(m，4H)，3.4(s，2H)，2.4(m，4H)；LRMS(ES+)m/z 340[MH]+
Example 7
4-amino-1-benzyl-2-oxo-2, 3-dihydro-1H-imidazo [4, 5, c ] pyridine-7-carboxylic acid cyclopropylmethyl-amide
1-benzyl-4-dibenzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5, c)]Pyridine-7-carboxylic acid cyclopropylmethyl-amide (10mg, 0.02mmol) was dissolved in 1ml of concentrated sulfuric acid and the mixture was stirred at room temperature for 30 minutes. Once complete, the mixture was diluted in 5ml of water and potassium carbonate was added in increments until pH-12. The mixture was then extracted with ethyl acetate (2X 50 ml). The organic layers were combined and MgSO4Dried and the solvent removed in vacuo. The residue was purified by silica gel column chromatography using 20% methanol in ethyl acetate to give 1mg of the title compound; LRMS (ES)+)m/z 338[MH]+
Example 8
4-amino-1-benzyl-7-bromo-6-methyl-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one
4-amino-1-benzyl-6-methyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one (20mg, 0.08mmol) was suspended in 5ml of acetic acid, followed by the addition of sodium acetate (5mg, 0.08mmol) and then the dropwise addition of bromine (4L, 0.08 mmol). The mixture was stirred at room temperature for 30 minutes. The mixture was diluted in water (50ml) and extracted with ethyl acetate (50ml), the organic layer was separated, dried over magnesium sulphate and the solvent removed in vacuo. The residue was purified by silica gel column chromatography using 10% methanol in ethyl acetate to give the title compound as a brown solid 15 mg.
1H NMR(d6 DMSO)δ：7.40-7.10(m，5H)，6.85(s，2H)，5.30(s，2H)，2.35(s，3H)；LRMS(ES+)m/z 333，335[MH]+
Example 9
4-amino-1-benzyl-6-methyl-5-oxy-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one
4-amino-1-benzyl-6-methyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one (20mg, 0.08mmol) was dissolved in 10ml of dichloromethane, then 3-chloroperoxybenzoic acid (15mg, 0.09mmol) was added and the mixture was stirred at room temperature for 2 hours. The mixture was washed with water, dried over magnesium sulfate and the solvent was removed in vacuo to yield 5mg of the title compound.
1H NMR(CD30D)δ：7.40-7.20(m，5H)，6.59(s，1H)，5.05(s，2H)，2.45(s，3H)；LRMS(ES+)m/z 271[MH]+
Example 10
4-amino-1-benzyl-6- (2-methoxy-ethyl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-benzylamino-6- (2-methoxy-ethyl) -1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (32mg, 0.082mmol) was stirred in concentrated sulfuric acid (2ml) for 30 minutes. Water (5ml) was added and the mixture was added dropwise to saturated NaHCO3To obtain an alkaline pH. The aqueous solution was extracted with 2 × EtOAc and the combined organics were dried and concentrated to give a yellow solid. With DCM: MeOH 97:3 (gradient increasing to DCM: MeOH: NH)395:5:0.5) and the mixture of isomers was separated by silica column chromatography to give the title compound as a pale yellow solid (3.1mg, 13%).
1H NMR(MeOD)δ 2.82-2.85(t，2H)，3.66-3.69(t，2H)，5.08(s，2H)，6.36(s，1H)，7.19-7.34(m，5H)；LRMS(ES)m/z 299[MH]+
Example 11
4-amino-1-benzyl-6- [2- (2-methoxy-ethylamino) -ethyl ] -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
Reacting [2- (1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ]]Pyridin-6-yl) -ethyl]- (2-methoxy-ethyl) -carbamic acid tert-butyl ester (64mg, 0.12mmol) was stirred in concentrated sulfuric acid (2ml) for 30 minutes. Water (5ml) was added and the mixture was added dropwise to saturated NaHCO3To obtain an alkaline pH. The aqueous solution was extracted with 2 × EtOAc and the combined organics were dried and concentrated to give a yellow solid. With DCM: MeOH 98:2 (gradient increasing to DCM: MeOH: NH)390:10:1) and the mixture of isomers was separated by silica column chromatography to give the title compound as a yellow solid (4.2mg, 11%).
1H NMR(MeOD)δ 2.84-2.86(t，2H)，3.02-3.05(t，2H)，3.27(s，3H)，3.45-3.48(t，2H)，5.09(s，2H)，6.34(s，1H)，7.20-7.34(m，5H)；LRMS(ES)m/z 342[MH]+
Example 12
4-amino-1-benzyl-6-oxazol-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
CDI (821mg, 5.06mmol) was added to N*2*，N*4*-dibenzyl-6-oxazol-2-yl-pyridine-2, 3, 4-triamine (940mg, 2.53mmol) in THF (15 ml). The solution was heated at 60 ℃ for 18 hours under nitrogen atmosphere. The reaction mixture was cooled and then concentrated in vacuo. The crude mixture was then dissolved in concentrated H 2SO4(15ml) and left to stand at room temperature for 30 minutes. The dark brown solution was added dropwise onto crushed ice. By adding K2CO3The saturated aqueous solution adjusted the pH to-9 and the mixture was filtered. The solid was washed with EtOAc (200ml) and the organics and aqueous filtrate were transferred to a separatory funnel. The layers were separated and the aqueous solution re-extracted with EtOAc (200 mL). The organics were combined and dried (MgSO)4) And evaporated to an orange sticky solid. The crude material was triturated with EtOAc and toluene. The resulting solid was filtered and washed with EtOAc to give an off-white solid. This material was purified by HPLC on a Phenomenex Gemini 5 μm column (150X 21.2mmid) eluting with 0.05% formic acid (aq) and 0.05% formic acid in MeCN at a flow rate of 15 ml/min. The gradient was isocratic at 5% organics for 0.6 min, then increased linearly from 5% to 80% organics over 12 min.
The filtrate from the trituration was evaporated and then subjected to column chromatography on an Isco company silica column (12g, Redisep). The resulting material was then eluted with EtOAc: MeOH, increasing the gradient linearly from 95:5 to 98:2 over 8 column volumes. The desired eluting components were combined and evaporated to an orange gummy solid. The material was purified by HPLC as above. All desired fractions eluted from the HPLC column were combined and evaporated to give the title compound as a white solid (26mg, 3%).
1H NMR(CD30D)δ 5.07(s，2H)7.17-7.36(m，7H)7.92(s，1H)LCMS Rt＝2.15m/z 308[MH]+
An alternative way of preparing example 12 is described below.
4-amino-1-benzyl-6-oxazol-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
At-78 ℃ under N2A solution of butyllithium in hexane (1.6M, 183. mu.l, 0.29mmol) was added dropwise under an atmosphere to a solution of oxazole (16. mu.l, 0.24mmol) in THF (1 ml). The solution was stirred at-78 ℃ for 10 minutes, then a solution of zinc chloride (100mg, 0.73mmol) in THF (1ml) was added dropwise. The solution was stirred at-78 ℃ for 15 minutes and then warmed to room temperature. The solution was then added to the solution containing 4-amino-1-benzyl-6-bromo-1, 3-dihydro-imidazo [4, 5-c ] using a syringe]Pyridin-2-one (13mg, 0.04mmol) and bis (triphenylphosphine) palladium dichloride (12mg, 0.02mmol) in a pre-sealed microwave vial (Biotage, 0.5-2.0ml) purged with nitrogen. The vial was heated at 110 ℃ for 15 minutes under microwave irradiation (Biotage, Initiator 8 (microwave synthesizer)). The reaction mixture was stirred in ethyl acetate (20ml) with saturated NH4Cl (aq) (10 ml). The mixture was filtered through Celite and washed with ethyl acetate (20 ml). The layers were separated and the organics were washed with water (10ml) and brine (10ml), MgSO4Dried and concentrated in vacuo to yield the crude product. The sample was dissolved in a mixture of acetonitrile: water: DMSO (2:1:1) and purified by preparative HPLC (FractionLynx) to give the title compound as a white solid (2 mg).
1H NMR(d6-DMSO)δ 10.60(brs，1H)，8.10(s，1H)，7.36-7.27(m，6H)，7.19(s，1H)，6.01(br s，2H)，5.04(s, 2H); LRMS (APCI and ES) m/z 308[ MH]+。
Example 13
4-amino-1-benzyl-6- (1-methyl-1H-imidazol-2-yl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
CDI (184mg, 1.13mmol) was added to N in the reaction flask*2*，N*4*-dibenzyl-6- (1-methyl-1H-imidazol-2-yl) -pyridine-2, 3, 4-triamine (218mg, 0.567mmol) in DMF (3 ml). The bottles were flushed with nitrogen, then sealed and heated in an aluminum block at 60 ℃ (aluminum block temperature). The dark brown solution was stirred at this temperature for 16 hours. The solution was concentrated under high vacuum and then dissolved in concentrated sulfuric acid (5 ml). The brown solution was stirred at room temperature for 30 minutes and then poured onto crushed ice (-20 ml). A saturated aqueous solution of potassium carbonate was added dropwise until pH-8. The aqueous solution was decanted from the solid precipitated during neutralization and then extracted with EtOAc (2 × 50 ml). The combined organics were dried (MgSO)4) Evaporated to a yellow solid (106 mg).
Samples (58mg) were chromatographed by HPLC on a Phenomenex Gemini 5 μm column (150X 21.2 mmid). Elution was performed with 0.05% DEA (aq) and 0.05% DEA in MeCN at a flow rate of 18 ml/min. The gradient was isocratic at 5% organics for 0.6 min, then increased linearly from 5% to 100% organics over 15 min.
The desired eluting components were combined and evaporated to give the title compound as a white solid (10mg, 6%).
1H NMR(CD30D)δ 3.94(s，3H)5.08(s，2H)6.91-6.95(m，1H)7.00(s，1H)7.07-7.10(m，1H)7.16-7.38(m，5H)；LRMS(ES+)m/z 321[MH]+
Example 14
4-amino-1-benzyl-6- (3-methyl- [1, 2, 4] oxadiazol-5-yl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c)]Pyridine-6-carboxylic acid (1-hydroxyimino-ethyl) -amide was suspended in toluene and sealed in a microwave bottle (Biotage, 0.5-2.0 ml). The vial was sealed and heated under microwave irradiation (Biotage Initiator 8) at 150 ℃ for 15 minutes. The sample was heated in a microwave at 150 ℃ for a further 30 minutes and then at 150 ℃ for a further 30 minutes. The mixture was evaporated and then partitioned between EtOAc (10ml) and water (5 ml). The aqueous solution was extracted twice more with EtOAc (2X 10mL) and the combined organics were dried (MgSO4) And is evaporated. The residue was suspended in acetonitrile (2ml) and the mixture was sealed in a microwave bottle and then heated under microwave irradiation at 170 ℃ for 30 minutes and then at 190 ℃ for a further 30 minutes. The reaction mixture was concentrated in vacuo and then dissolved in concentrated H2SO4(2 ml). The solution was stirred for 30 minutes and then poured onto crushed ice. Saturated K is added dropwise 2CO3The aqueous solution was brought to pH-8. The aqueous solution was decanted from the solid into a separatory funnel and then extracted with EtOAc (3 × 15 ml). The combined organics were dried (over MgSO4) And is evaporated. The isomer mixture was purified by HPLC on a Luna 10 micron C18(2) column (150 × 21.2 mmid). Elution was performed with 0.1% formic acid (aq) and 0.1% formic acid in MeCN at a flow rate of 25 ml/min. The gradient was isocratic at 5% organics for 0.6 min, then increased linearly from 5% to 90% organics over 8.50 min. The desired fractions were evaporated to give the title compound as a white solid (0.5mg, 1%).
LCMS Rt＝2.46m/z 323[MH]+。
Example 15
4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-benzylamino-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (2.63g, 6.60mmol) was dissolved in concentrated sulfuric acid (50ml) and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was cooled to 0 ℃ and ice was added. Will K2CO3(150g) Dissolved in water (700ml) and the reaction mixture was added dropwise. The aqueous solution was extracted with EtOAc (6X 500 mL). The combined organics were washed with brine (200ml), MgSO4Dried and concentrated in vacuo to give the title compound as a white solid (1.20 g).
1H NMR(CDCl3) δ 10.77(brs, 1H), 7.36-7.25(m, 5H), 7.01(s, 1H), 6.20 (brs, 2H), 5.04(s, 2H); LRMS (APCI and ES) m/z 309[ MH]+，307[MH]-. Found C.54.55 H.3.60 N.18.17%. C14H11F3N4O requires% C.54.36H.3.61 N.17.86.
An alternative synthesis to example 15 is described below;
ethyl- [2, 3-diamino-6- (trifluoromethyl) -pyridin-4-yl ] -benzylcarbamate (35gm, 99mmol) was dissolved in glacial acetic acid (300ml) at room temperature. Any insoluble material was removed by filtration and the clear yellow filtrate was then heated to 80 ℃ with stirring. A white precipitate began to form within 10 minutes. Heating was continued for a total of 40 minutes. The reaction mixture was cooled to ambient temperature and the precipitate was collected by filtration, washed with acetic acid and dried in vacuo at 50 ℃ for 3 hours to yield the title compound as a white solid (26.4gm, 86% yield).
Example 16
4-amino-1- (6-methyl-pyridin-3-ylmethyl) -6-oxazol-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
CDI (272mg) was added to a solution of N2, N2-dibenzyl-N4- (6-methyl-pyridin-3-ylmethyl) -6-oxazol-2-yl-pyridine-2, 3, 4-triamine (400mg) in THF (10 ml). The solution is placed in N2Stirred at 60 ℃ for 5 hours under an atmosphere. An additional 3 equivalents of CDI (408mg) were added and the reaction heated at reflux for 16 h. The reaction mixture was cooled and then concentrated in vacuo. The crude mixture was then dissolved in concentrated H 2SO4(5ml) and left to stand at room temperature for 30 minutes. The dark brown solution was added dropwise onto crushed ice. By adding K2CO3The saturated solution adjusted the pH to-8 and the mixture was filtered. The resulting solid was washed with EtOAc (50ml) and the organics and aqueous filtrate were transferred to a separatory funnel. The layers were separated and the aqueous solution re-extracted with EtOAc (50 mL). The organics were combined and dried (over MgSO)4) And evaporated to produce a sticky solid. The mixture was washed with Et2Triturate with O, and collect the solid by filtration and use Et2Wash O yielded the title compound as an off-white solid (30 mg).
1H NMR(CDCl3，400MHz)δ 2.45(s，3H)，5.09(s，2H)，7.21-7.28(m，2H)，7.30(s，1H)，7.70(dd，1H)，7.95(s，1H)，8.42(d，1H)。LCMS Rt＝2.29m/z 323[MH]+
Example 17
4-amino-1-benzyl-6- (2-methoxy-ethoxy) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
[ 2-amino-6- (2-methoxy-ethoxy) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester (118mg) was dissolved in ethanol (5ml) and 10% Pd on carbon (15mg) was added. The reaction was stirred under hydrogen atmosphere (50psi) at room temperature for 1 hour. The reaction mixture was then filtered through a pad of celite and evaporated. The crude product was dissolved in glacial acetic acid (2ml) and transferred to a microwave vial (Biotage, 0.5-2.0 ml). The vial was sealed and heated under microwave irradiation at 100 ℃ for 5 minutes. The resulting brown mixture was evaporated, the solid was loaded on silica and column chromatographed on an isco company silica column (4g, Redisep) eluting with EtOAc: heptane elution (linearly increasing the gradient from 60: 40 to 100% EtOAc over 8 Column Volumes (CVs) and then isocratic for 8 CVs at 100% EtOAc. The desired eluting components were combined and evaporated to give the title compound as a white solid (50 mg).
1H NMR(CDCl3，400MHz)δ 3.35(s，3H)，3.62(t，2H)，4.20(t，2H)，4.95(s，2H)，5.80(s，1H)，7.20-7.38(m，5H)。LCMS Rt＝2.13m/z 315[MH]+
Example 18
4-amino-1- (6-methyl-pyridin-3-ylmethyl) -6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
(2-amino-3-nitro-6-trifluoromethyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (27mg, 0.07mmol) was stirred in glacial acetic acid (5ml) and the solution was stirred at ambient temperature at 40psi H2Stirred under an atmosphere in the presence of 10% Pd/C (5.4mg, 20% wt) for 4 hours. The suspension was filtered through an Arbocel pad, washed with 2 × 3ml AcOH and the filtrate was concentrated in vacuo. Acetonitrile (5ml) was added to the residue and the material was triturated and the resulting solid removed by filtration and dried in vacuo to give the title compound as a white solid.
1H NMR(d6-DMSO)：δ 8.45(s，1H)，7.57-7.55(d，1H)，7.20-7.18(d，1H)，7.10(s，1H)，6.20(bs，2H)，5.01(s，2H)，2.40(s，3H)；LRMS(APCI)m/z 324[MH]+
An alternative way of preparing example 18 is described below.
4-amino-1- (6-methyl-pyridin-3-ylmethyl) -6-trifluoromethyl-1, 3-dihydro-imidazo (4, 5-c) pyridin-2-one
4-benzylamino-1- (6-methyl-pyridin-3-ylmethyl) -6-trifluoromethyl-1, 3-dihydro-imidazo (4, 5-c) pyridin-2-one (30mg, 0.07mmol) was dissolved in 1ml of concentrated sulfuric acid and the reaction mixture was stirred at room temperature for 1 hour. The mixture was poured into water (100ml) and potassium carbonate was added in succession until the pH was basic. Then extracted with ethyl acetate (100 ml). The organic layer was separated, dried over magnesium sulfate and the solvent was removed in vacuo to yield 16mg of the title compound as a white solid.
1H NMR(d6-DMSO)：δ 8.45(s，1H)，7.57-7.55(d，1H)，7.20-7.18(d，1H)，7.10(s，1H)，6.20(bs，2H)，5.01(s，2H)，2.40(s，3H)；LRMS(APCI)m/z 324[MH]+
Example 19
4-amino-6- (4-methyl-oxazol-2-yl) -1- (6-methyl-pyridin-3-ylmethyl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
A mixture of raney nickel (5mg) and [ 2-amino-6- (4-methyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (69mg) in acetic acid (3ml) was stirred under a hydrogen atmosphere (80psi) for 1 hour. Arobcel was added on top of a sulfonic acid cation exchange cartridge (Bakerbond, 1g) and the reaction mixture was loaded on the top for filtration. The catalyst and Arbocel were removed with a spatula and the cartridge was then washed with methanol (5ml) to remove impurities. The product was released from the cartridge by eluting with ammonia (2M, 2 × 5ml) in methanol. The crude solution was evaporated, then IPA (3ml) was added, the solid precipitated, collected by filtration and washed with IPA. The resulting off-white solid was dried under high vacuum to give the title compound (16 mg).
1H NMR(400MHz，DMSO-d6)δ ppm 2.13(s，3H)2.42(s，3H)5.05(s，2H)5.98(s，2 H)7.21(d，J＝7.90Hz，1 H)7.25(s，1 H)7.56(dd，J＝7.90，2.44Hz，1 H)7.80(s，1 H)8.47(d，J＝2.44Hz，1 H)10.68(br.s.，1H)；LRMS(ESI)m/z 337[MH]+，335[MH]-
Example 20
4-amino-6- (4-ethyl-oxazol-2-yl) -1- (6-methyl-pyridin-3-ylmethyl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
[ 2-amino-6- (4-ethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (74mg) was dissolved in acetic acid (2ml) and zinc powder (113mg, Aldrich, 99%) was added to the reaction. The mixture was stirred at room temperature under nitrogen atmosphere for 16 hours. The reaction mixture was filtered directly on a cation exchange cartridge (Bakerbond SCX, sulfonic acid binder phase, 1 g). The SCX cartridge was washed with methanol (2 x 4ml) to remove impurities and then the product was liberated with ammonia in methanol (2M, 4 ml). The desired eluting components were combined and evaporated to an off-white solid, triturated with isopropanol, filtered, then washed with isopropanol to give the title compound as a white solid (32 mg).
1H NMR(400MHz，DMSO-d6)δ ppm 1.16(t，J＝7.42Hz，3 H)2.40(s，3 H)2.44-2.56(m，2 H)5.04(s，2 H)5.98(s，2 H)7.19(d，J＝8.02Hz，1 H)7.23(s，1 H)7.54(dd，J＝8.02，2.34Hz，1 H)7.79(s，1 H)8.45(d，J＝2.34Hz，1 H)10.65(s，1 H)；LCMS Rt＝1.73m/z 351[MH]+
Example 21
4-amino-1-benzyl-6- (1H-imidazol-2-yl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
The title compound was prepared according to the method of example 20 using { 2-amino-3-nitro-6- [1- (2-trimethylsilyl-ethoxymethyl) -1H-imidazol-2-yl ] -pyridin-4-yl } -benzyl-carbamic acid ethyl ester (174mg) and raney nickel (5mg) in acetic acid (3 ml). This resulted in the initial production of SEM protected imidazole compounds. Hydrogen chloride in dioxane (4M, 1ml) was then added dropwise and the solution was stirred at room temperature for 24 hours. The reaction mixture was then transferred to a microwave vial (Biotage, 2-5ml) and heated under microwave irradiation (Biotage Initiator 8) at 110 ℃ for 10 minutes. The reaction mixture was evaporated and then redissolved in methanol, and the solution was loaded on a cation exchange cartridge (Bakerbond, sulfonic acid binder phase, 1 g). The cartridge was washed with methanol (2 x 5ml) to remove impurities and then the product was liberated by elution with ammonia in methanol (2M, 5 ml). The desired eluting components were combined and evaporated to a brown solid. This was triturated with isopropanol and the solid collected by filtration then washed with more isopropanol to give the title compound as a pale brown solid (28 mg).
1H NMR(400MHz，DMSO-d6)δ ppm 5.02(s，2 H)5.58(s，2 H)6.91(s，1H)7.06(s，1 H)7.11(s，1 H)7.25-7.36(m，5 H)10.53(s，1 H)11.99(s，1 H)；LCMS Rt＝1.52m/z 307[MH]+。
Example 22
4-amino-1-benzyl-6- (2-fluoro-phenyl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
[ 2-amino-6- (2-fluoro-phenyl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester (31mg) was dissolved in acetic acid (1 ml). Zinc powder (Aldrich, 99%, 20mg) was added and the mixture was stirred at room temperature under nitrogen for 2 hours. Additional zinc powder (30mg) was added and the mixture was stirred for an additional 1 hour. The reaction mixture was diluted with methanol (2ml) and then filtered directly on a cation exchange cartridge (Bakerbond SCX, sulfonic acid bound phase, 1 g). The SCX cartridge was washed with methanol (2 x 5ml) to remove impurities and then the product was liberated with ammonia in methanol (2M, 5 ml). The desired eluate fractions were evaporated to a light brown solid. This was triturated with ethyl acetate then filtered and washed with more ethyl acetate to give the title compound as a pale purple solid (8 mg).
1H NMR(400MHz，DMS0-d6)δ ppm 4.99(s，2 H)5.82(s，2 H)6.87(d，J＝1.95Hz，1 H)7.15-7.29(m，3 H)7.29-7.39(m，5 H)7.81(td，J＝8.01，1.95Hz，1 H)10.51(s，1 H)；LCMS Rt＝2.21m/z 335[MH]+
The following examples 23-119 can be prepared in a similar manner to examples 1-22 from similar intermediates described in the preparative section using similar chemistry.
Examples 23-28 were prepared according to the method of example 1, examples 54-60, 65-72 and 107-110 were prepared according to the method of examples 15 and 18, examples 31 and 61 were prepared according to the method of example 3, examples 42-49, 83-95 and 101-102 were prepared according to the method of example 17, examples 73-78, 96-99, 103-106 and 111-112 were prepared according to the method of example 19, examples 79-81 were prepared according to the method of example 22, examples 34-35 and 40 were prepared according to the method of example 6, examples 29-30, 36-39, 41 and 50-53 were prepared according to the method of example 4, examples 32-33 were prepared according to the method of example 5, prepared as described in example 14.
Examples 62-64 were prepared as described in examples 15 and 18, example 82 as described in example 14, and example 100 as described in example 17.
In the following example tables, asterisks indicate attachment points.
Preparation 1
3-amino-3-cyclopropyl-acrylic acid ethyl ester
Cyclopropionitrile (2.7g, 40.5mmol) was dissolved in anhydrous THF (100ml), then zinc (13.2g, 202.3mmol) and zinc oxide (1.6g, 20.2mmol) were added, then ethyl bromoacetate (6.7g, 40.5mmol) was added dropwise. Mixing the mixture in N2Sonicate for 2 hours in a 35kHz ultrasonic bath under an atmosphere. After 30 minutes a green colour was observed. The mixture was filtered through celite to remove zinc and zinc oxide. Adding the filtrate to 2050% K of ml2CO3(aq) in solution. The thick precipitate formed was filtered to remove solids and the aqueous solution was extracted with 100ml EtOAc. The extract was washed with 20ml brine, MgSO4Dried and concentrated in vacuo to give the crude product (3.8 g). The reaction mixture was purified using 90:10 pentane: the crude material was purified by silica gel column chromatography eluting with EtOAc to give the title compound as a yellow oil (1.32 g).
1H NMR (CDCl3) delta 4.47(s, 1H), 4.11 (quartet, 2H), 1.45-1.39(m, 1H), 1.26(t, 3H), 0.88-0.83(m, 2H), 0.76-0.72(m, 2H); LRMS (APCI +) m/z156 MH ]+。
Preparation 2
6-cyclopropyl-2, 4-dihydroxy-nicotinic acid ethyl ester
Sodium metal (8.1g, 119mmol) was cut into small pieces and added to stirred ethanol (120ml) successively at room temperature under nitrogen. The mixture was then incubated at 60 ℃ under N2Stir overnight under atmosphere to ensure complete dissolution of the metal. Diethyl malonate (18.1ml, 119mmol) was added to the sodium ethoxide solution at 60 ℃ and the mixture was stirred under N2Stirred at 60 ℃ for 1 hour under an atmosphere. A solution of 3-amino-3-cyclopropyl-acrylic acid ethyl ester (10.3g, 40mmol) in ethanol (10ml) was added dropwise at 60 ℃ and the mixture was refluxed at N2Heat under atmosphere for 5 days to give an orange suspension. The mixture was cooled to room temperature and the resulting solid was collected by filtration. The filtrate was concentrated in vacuo to yield more solid. The combined solids were dissolved in water (150ml) and the solution was washed with EtOAc (150 ml). The aqueous solution was acidified to pH 2 using concentrated HCl, resulting in precipitation of a white solid. The solid was collected by filtration and then washed with cold water and Et2O wash, then dry in vacuo at 40 ℃ overnight to yield the title product as a fine white solid (5.32 g). The filtrate was concentrated in vacuo to half its volume, precipitating more product. The solid from this 2 nd harvest was collected by filtration, washed with water and Et 2O washed and dried in vacuo at 40 ℃ to yield another 0.35g of the title compound as a beige solid.
1H NMR (d 6-DMSO). delta.12.71 (br s, 1H), 11.43(br s, 1H), 5.51(s, 1H), 4.26 (quartet, 2H), 1.86-1.79(m, 1H), 1.26(t, 3H), 1.06-1.01(m, 2H), 0.90-0.86(m, 2H); LRMS (APCI) m/z 224[ MH]+。
Preparation 3
6-cyclopropyl-2, 4-dihydroxy-pyridines
Ethyl 6-cyclopropyl-2, 4-dihydroxy-nicotinate (5.3g, 20.5mmol) was dissolved in concentrated HCl (25ml) and the mixture was refluxed overnight. The mixture was cooled to room temperature and then neutralized with concentrated ammonia. The resulting precipitate was collected by filtration, washed with cold water and acetonitrile and dried in vacuo at 40 ℃ for 2 days to give the title compound (3.39g) as a beige powder.
1H NMR(d6-DMSO)δ 10.96(br s，1H)，10.25(br s，1H)，5.38，(d，1H)，5.32(d，1H)，1.79-1.72(m，1H)，0.93-0.89(m，2H)，0.75-0.71(m，2H)；LRMS(ES)m/z 152[MH]+。
Preparation 4
6-cyclopropyl-2, 4-dihydroxy-3-nitro-pyridines
6-cyclopropyl-2, 4-dihydroxy-pyridine (1g, 6.6mmol) was suspended in AcOH: EtOAc (4:1, 10ml) at room temperature. The mixture was warmed to 30 ℃ and a small portion of fuming nitric acid (0.05ml, 1.2mmol) was added dropwise, maintaining the temperature between 30 ℃ and 35 ℃. The mixture became a clear solution after addition. The remaining fuming nitric acid (0.25ml, 6.3mmol) was added dropwise. The clear solution was cooled to room temperature whereupon a precipitate began to form. The mixture was stirred at room temperature overnight. The solid was collected by filtration, washed with cold water and Et 2O washed and dried in vacuo at room temperature over another week (one weekend) to give the title compound (1.21g) as a yellow powder.
1H NMR(d6-DMSO)δ 12.17(br s，1H)，11.88(br s，1H)，5.57(s，1H)，1.88-1.81(m，1H)，1.08-1.03(m，2H)，0.87-0.83(m，2H)；LRMS(APCI)m/z 197[MH]+。
Preparation 5
6-cyclopropyl-2, 4-dichloro-3-nitro-pyridine
6-cyclopropyl-2, 4-dihydroxy-3-nitro-pyridine (1.2g, 6.1mmol) was suspended in POCl3(5 mL). The mixture was heated under a caustic scrubber at 85 ℃ overnight. Adding excessive POCl3The reaction residue was removed in vacuo, dissolved in EtOAc (50ml) and added dropwise to stirred warm water (50ml) at controlled temperature using ice. The layers were separated and the aqueous solution was extracted with 90:10 EtOAc: MeOH (100 ml). The organics were washed with brine (50ml), MgSO4Dried and concentrated to give the crude product (2 g). Elution with 90:10 pentane: EtOAc and chromatography on a silica gel column gave the title compound as a pale yellow crystalline solid (893 mg).
1H NMR(d6-DMSO)δ 7.94(s，1H)，2.31-2.24(m，1H)，1.18-1.14(m，2H)，1.06-1.02(m，2H)；LRMS(APCI)m/z 233[MH]+。
Preparation 6
Benzyl- (2-chloro-6-cyclopropyl-3-nitro-pyridin-4-yl) -amine
6-cyclopropyl-2, 4-dichloro-3-nitro-pyridine (160mg, 0.8mmol) was dissolved in THF (2ml), and triethylamine (1041, 0.8mmol) and benzylamine (811, 0.8mmol) were added. The mixture was stirred at room temperature under nitrogen for 48 hours, and then a yellow precipitate formed. The volatiles were removed in vacuo and the residue was stored in a stoppered flask at room temperature for 10 days. The residue was then purified by silica gel column chromatography eluting with 99:1 DCM: MeOH, 98:2 DCM: MeOH to give the title compound (185mg) as a yellow crystalline solid.
1H NMR(CDCl3)δ 7.42-7.31(m，5H)，7.05(br s，1H)，6.47(s，1H)，4.49(d，2H)，1.88-1.81(m，1H)，1.09-1.04(m，2H)，1.01-0.96(m，2H)；LRMS(APCI)m/z 304[MH]+。
Preparation 7
Benzyl- (3-amino-2-chloro-6-cyclopropyl-pyridin-4-yl) -amine
Benzyl- (2-chloro-6-cyclopropyl-3-nitro-pyridin-4-yl) -amine (245mg, 0.8mmol) was dissolved in AcOH H2O (9.0:0.9 ml). Iron powder (270mg, 4.8mmol) was added and the mixture was stirred vigorously at room temperature under nitrogen for additional weeks during which time an off-white precipitate precipitated out. The reaction mixture was diluted with EtOAc (20ml) and water (20ml), the mixture was filtered through celite and the filter cake was washed with EtOAc (20 ml). The phases were separated and the organic layer was washed with saturated NaHCO3The aqueous solution (10ml) and brine (10ml) were washed with MgSO4Dried and concentrated in vacuo. The residue was dried in vacuo at 40 ℃ overnight to give the title compound as an off-white crystalline solid (215 mg).
1H NMR(CDCl3)δ 7.40-7.31(m，5H)，6.29(s，1H)，4.59(br s1H)，4.37(d，2H)，3.30(br s 2H)，1.89-1.82(m，1H)，0.87-0.86(m，4H)；LRMS(APCI)m/z 274[MH]+。
Preparation 8
1-benzyl-4-chloro-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c]Pyridin-2-one benzyl- (3-amino-2-chloro-6-cyclopropyl-pyridin-4-yl) -amine (210mg, 0.8mmol) was dissolved in acetonitrile (10 ml). 1, 1-carbonyldiimidazole (370mg, 2.3mmol) was added and the mixture was heated at 80 ℃ for 2 hours under nitrogen atmosphere. A further 250mg (1.5mmol) of 1, 1-carbonyldiimidazole are added and the mixture is heated at 80 ℃ overnight. The mixture was cooled to room temperature and the solvent was removed in vacuo. The residue was dissolved in DCM (20ml) and washed with 1N HCl (10ml), water (10ml) and brine (10ml), MgSO 4Dried and concentrated in vacuo. The residue was dried in vacuo at 40 ℃ overnight to give the title compound as a white fluffy solid (217 mg).
1H NMR(CDCl3)δ 8.20(brs，1H)，7.38-7.31(m，5H)，6.59(s，1H)，5.03(s，2H)，1.96-1.91(m，1H)，0.94-0.92(m，4H)；LRMS(APCI)m/z300[MH]+。
Preparation 9
4-allylamino-1-benzyl-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
1-benzyl-4-chloro-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c)]Pyridin-2-one (100mg, 0.3mmol) was dissolved in reaction vial reactiveTMAmong allylamine (2mL) in (1). Copper (II) sulfate (83mg, 0.3mmol) was added and the bottle was sealed. The mixture was heated at 85 ℃ overnight. More copper (II) sulfate (83mg, 0.3mmol) and allylamine (1ml) was added and the bottle was sealed again. The mixture was heated at 85 ℃ for weeks. The mixture was cooled to room temperature. The excess allylamine was removed in vacuo and the residue was dissolved in EtOAc (50mL) and saturated NaHCO3Aqueous solution (20ml) was treated. The layers were separated and the organics were reused with more saturated NaHCO3The aqueous solution (10ml), brine (10ml) were washed with MgSO4Dried and concentrated in vacuo to give the crude product (120 mg). Column chromatography on silica gel eluting with 98:2 DCM: MeOH gave the title compound as an off-white solid (73 mg).
1H NMR(CDCl3)δ 10.40(br s，1H)，7.34-7.25(m，5H)，6.17-6.15(m，1H)，5.96-5.87(m，1H)，5.17(d，1H)，5.02-5.00(m，1H)，4.89(s，2H)，4.05-4.00(m，2H)，1.85-1.80(m，1H)，0.97-0.93(m，2H)，0.84-0.76(m，2H)；LRMS(APCI)m/z 321[MH]+。
Preparation 10
Benzyl- (2-chloro-6-methyl-3-nitro-pyridin-4-yl) -amine
2, 4-dichloro-6-methyl-3-nitro-pyridine (2g, 9.7mmol) and triethylamine (1.35mL, 9.7mmol) were dissolved in 40mL THF and cooled (ice/water) to-5 ℃. A solution of benzylamine (1.04g, 9.7mmol) in 10ml THF was added dropwise and the mixture was then gradually warmed to room temperature overnight. The mixture was evaporated in vacuo and partitioned between EtOAc (50ml) and water (20 ml). The organic layer was washed with saturated NaHCO 3The aqueous solution (10ml) was washed with MgSO4Dried and evaporated in vacuo to an orange colloid. This colloid was pre-adsorbed on silica gel and then purified by column chromatography eluting with DCM, pentane 3: 1. The appropriate fractions were combined and evaporated in vacuo to give the title compound as a yellow solid (716 mg).
1H NMR(CDCl3) δ 2.32(s, 3H), 4.38(d, 2H), 6.39(s, 1H), 6.90 (broad singlet, 1H), 7.21(m, 2H), 7.29(m, 3H). LC-MS (ELSD, ES)+)m/z 278(MH+)。
Preparation 11
N-2 ', N-2 ', N-4 ' -tribenzyl-6-methyl-3-nitro-2, 4-diamine
Benzyl- (2-chloro-6-methyl-3-nitro-pyridin-4-yl) -amine (99mg, 0.4mmol) and triethylamine (551, 0.4mmol) were dissolved in THF (2ml) and dibenzylamine (77mg, 0.4mmol) was added dropwise. The resulting reaction mixture was stirred at room temperature overnight and then evaporated in vacuo. The residue was taken up in EtOAc (5ml) and saturated NaHCO3The aqueous solution (3ml) was partitioned between. The organic layer was dried (over MgSO4) And evaporated in vacuo to a yellow colloid, which was pre-adsorbed onto silica gel and then purified by column chromatography eluting with 1:1 DCM: pentane. The appropriate fractions were combined and evaporated in vacuo to a bright yellow colloid which solidified on standing to give the title compound (75 mg).
1H NMR(CDCl3) δ 2.32(s, 3H), 4.45(d, 2H), 4.54(s, 4H), 5.96(s, 1H), 7.13-7.40(m, 15H), 8.12 (broad singlet, 1H). LRMS (ES)+)m/z 439(MH+)。
Preparation 12
N-2 ', N-2 ', N-4 ' -tribenzyl-6-methyl-2, 3, 4-triamine
N-2 ', N-2 ', N-4 ' -tribenzyl-6-methyl-3-nitro-2, 4-diamine (59mg, 0.14mmol) was dissolved in ethanol (5ml) and hydrogenated over Raney nickel (6mg) at 30psi for 1 hour at room temperature. 12mg of Raney nickel were added and the mixture was hydrogenated at 30psi and room temperature for a further 1.5 hours. The reaction mixture was filtered through a short plug of Arbocel and the filtrate was evaporated in vacuo to give the title compound as an opaque colloid (39 mg).
1H NMR (DMSO). delta.2.08 (s, 3H), 4.07(s, 4H), 4.21(s, 2H), 4.30(d, 2H), 5.83(t, 1H interchangeable), 6.05(s, 1H), 7.14-7.34(m, 15H); LRMS (APCI)+)m/z409(MH+)。
Preparation 13
1-benzyl-4-dibenzylamino-6-methyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
N-2 ', N-2 ', N-4 ' -tribenzyl-6-methyl-2, 3, 4-triamine (35mg, 0.09mmol) and 1, 1-carbonyldiimidazole (139mg, 0.86mmol) were dissolved in acetonitrile (3ml) and the mixture was heated at reflux for 3 hours. The reaction mixture was evaporated in vacuo and the residue was purified by column chromatography using DCM as eluent. The appropriate fractions were combined and evaporated in vacuo to give the title compound as a white solid (30 mg).
1H NMR (CDCl3) Δ 2.32 (broad singlet, 3H), 4.68(s, 4H), 4.85(s, 2H), 6.18(s, 1H), 7.18-7.26(m, 15H); LRMS (ES)+)m/z 435(MH+)。
Preparation 14
N4-benzyl-2-chloro-6-trifluoromethyl-pyridine-3, 4-diamine
Benzyl- (2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) -amine (345mg, 1.0mmol) was dissolved in a mixture of AcOH (18ml) and water (2 ml). Iron powder (349mg, 6.2mmol) was added and the mixture was stirred vigorously at room temperature for 24 hours. The reaction mixture was concentrated in vacuo and the residue diluted with EtOAc (10ml) and water (10 ml). The mixture was filtered through celite, washing with EtOAc (20 ml). The layers were separated and the organic layer was washed with saturated NaHCO3The aqueous solution (2X 10ml) and brine (10ml) were washed with MgSO4Dried and concentrated in vacuo to give a pale yellow colorTitle compound as a solid (304 g).
1H NMR(CDCl3) δ 7.43-7.34(m, 5H), 6.87(s, 1H), 4.46(br s, 1H), 4.42(d, 2H), 3.72(br s, 2H); LRMS (APCI and ES) m/z 302[ MH]+。
Preparation 15
1-benzyl-4-chloro-6-trifluoromethyl-pyridin-2-one
N4-benzyl-2-chloro-6-trifluoromethyl-pyridine-3, 4-diamine (300mg, 1.0mmol) was dissolved in MeCN (20 ml). CDI (806mg, 4.9mmol) was added and the mixture was heated at 80 ℃ for 48 h. The mixture was cooled to room temperature and the solvent was removed in vacuo. The residue was dissolved in EtOAc (50mL) and washed with 1N HCl (aq) (20mL), water (20mL) and brine (20mL), MgSO 4Dried and concentrated in vacuo to give the title compound (325g) as a pale yellow solid.
1H NMR(CDCl3) δ 7.41-7.33(m, 5H), 7.14(s, 1H), 5.11(s, 2H); LRMS (APCI and ES) m/z 328[ MH ]]+。
Preparation 16
1-benzyl-4-benzylamino-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
Dissolve 1-benzyl-4-chloro-6-trifluoromethyl-pyridin-2-one (100mg, 0.3mmol) in BnNH in a reaction flask2(2 ml). Adding CuSO4(152mg, 0.6mmol) and the vial sealed. The reaction mixture was heated at 80 ℃ for 120 hours. The reaction mixture was then cooled to room temperature and dissolved in EtOAc (20 ml). The mixture was washed with saturated NaHCO3The aqueous solution (2X 5ml) and brine (5ml) were washed with MgSO 44Dried and concentrated in vacuo to give the crude product (700 mg). Elution with 99:1DCM: MeOH followed by silica column chromatography gave the title compound (50mg) as a yellow solid.
1H NMR(CDCl3)δ 10.66(br s，1H)，7.34-7.10(m，10H)，6.58(s，1H)，5.86-5.84(m，1H)，4.70(d, 2H), 4.66(s, 2H); LRMS (APCI and ES) m/z 399[ MH ]]+。
Alternative preparations of the above title compound are described below;
n2, N4-dibenzyl-6-trifluoromethyl-pyridine-2, 3, 4-triamine (9.02g, 24.2mmol) was dissolved in TBME (180ml), and CDI (19.6g, 121mmol) was added. The reaction mixture was stirred at room temperature for 72 hours. Water (100ml) was added to the reaction mixture and the layers were separated. The aqueous solution was extracted with EtOAc (200 mL). The combined organics were washed with brine (50ml), MgSO 4Dried and concentrated in vacuo to give the crude product (25 g). Elution with 30:70 → 60:40 heptane: EtOAc gave the title compound as a white fluffy solid by column chromatography on silica (2.64 g).
1H NMR(CDCl3) δ 10.52(br s, 1H), 7.44-7.12(m, 10H), 6.60(s, 1H), 5.76-5.72(m, 1H), 4.71-4.70(m, 4H); LRMS (APCI and ES) m/z 399[ MH ]]+。
Preparation 17
N2, N4-dibenzyl-3-nitro-6-trifluoromethyl-pyridine-2, 4-diamine
3-Nitro-6-trifluoromethyl-pyridine-2, 4-diol (5.0g, 22.3mmol) was dissolved in DCM (50mL) and Et was added3N (6.22ml, 44.6 mmol). The mixture was cooled to 0 ℃ and Tf was added dropwise2O (7.32ml, 44.6 mmol). The mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was concentrated in vacuo and the residue was dissolved in THF (50 ml). Addition of BnNH2(7.3ml, 66.9mmol) and the mixture was stirred at 50 ℃ for 24 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was treated with water (50ml) and extracted with EtOAc (150 ml). The extract was washed with brine (50ml), using MgSO4Dried and concentrated in vacuo to give the crude product (27 g).
A second crop of 3-nitro-6-trifluoromethyl-pyridine-2, 4-diol (11.06g, 49.4mmol) was dissolved in DCM (100mL) and Et was added 3N (13.8ml, 98.7 mmol). Cooling the mixture to 0 deg.CAnd Tf was added dropwise2O (16.2ml, 98.7 mmol). The mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was concentrated in vacuo and the residue was dissolved in THF (100 ml). Addition of BnNH2(16.2ml, 148mmol) and the mixture was stirred at 50 ℃ for 24 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was treated with water (100ml) and extracted with EtOAc (200 ml). The extract was washed with brine (50ml), using MgSO4Dried and concentrated in vacuo to give the crude product (53 g). The two crude products were combined. Elution with 95:5 → 90:10 pentane: EtOAc provided the title compound as a yellow solid by column chromatography on silica (15.93 g).
1H NMR(CDCl3) δ 9.68-9.64(m, 1H), 9.36-9.32(m, 1H), 7.43-7.29(m, 10H), 6.38(s, 1H), 4.81(d, 2H), 4.55(d, 2H); LRMS (APCI and ES) m/z 403[ MH]+。
Preparation 18
N2, N4-dibenzyl-6-trifluoromethyl-pyridine-2, 3, 4-triamine
N2, N4-dibenzyl-3-nitro-6-trifluoromethyl-pyridine-2, 4-diamine (15.9g, 35.6mmol) was dissolved in a mixture of THF (100ml) and MeOH (200 ml). Raney nickel (3.18g, 20 wt%) was added and the mixture was allowed to stand at 80psi H at room temperature2Stirred for 1 hour. The mixture was filtered through celite to remove the catalyst and the filtrate was concentrated in vacuo to give an oil. Trituration with a small amount of water in MeOH yielded a precipitate, which was collected by filtration, washed with cold MeOH and dried in vacuo to yield the title compound as a white solid (9.02 g).
1H NMR(CDCl3) δ 7.43-7.28(m, 10H), 6.57(s, 1H), 4.66(d, 2H), 4.62-4.59(m, 1H), 4.57-4.54(m, 1H), 4.39(d, 2H), 2.49(br s, 2H); LRMS (APCI and ES) m/z 373[ MH)]+。
Preparation 19
2, 4-dihydroxy-6-trifluoromethyl-nicotinic acid ethyl ester
Pyridine (53ml/660mmol) was added to dissolve ethyl 3-amino-4, 4, 4-trifluorocrotonate (100g/546mmol) in DCM (6000 ml). The mixture was then placed under nitrogen and cooled to 5 ℃ by suspension in an ice bath. Ethylmalonyl chloride was added dropwise over about 1 hour so that the temperature did not exceed 20 ℃. The resulting light brown solution was stirred at 5 ℃ for 3 hours and then warmed to room temperature overnight to give a dark green solution. The mixture was then washed with 1M aqueous HCl (200ml), saturated NaHCO3Aqueous solution (250ml) was washed. The aqueous washes were further re-extracted with additional DCM (2X 250 ml). The organic layers were combined and washed with Na2SO4Dried, filtered and concentrated to crude 3- (2-ethoxycarbonyl-acetylamino) -4, 4, 4-trifluoro-but-2-enoic acid ethyl ester (175g) as a dark green oil. A portion of the crude material (120g) was dissolved in EtOH (300ml) and placed under a nitrogen atmosphere. Potassium tert-butoxide (54g/480mmol) was then added in portions so that the temperature did not exceed 60 ℃ to give a violet solution. The mixture was then heated at 70 ℃ for 3 hours. EtOH (100ml) was then added to reduce the viscosity and heated at 80 ℃ for an additional 1 hour. The mixture was then cooled and then concentrated in vacuo to a red solid. The mixture was dissolved in water (500ml), and citric acid (180g) was added to produce a precipitate. EtOAc (600ml) was then added and the mixture poured into a separatory funnel and the aqueous layer drained. The organic layer containing a large amount of insoluble solids was filtered to give the title compound as a white solid (46.5 g). The organic filtrate was concentrated and triturated with MeOH to give more title compound as a white solid (15.3 g).
1H NMR(d6-DMSO，400MHz)δ 1.20-1.25(t，3H)，4.20-4.25(q，2H)，6.8(s，1H)
Preparation 20
6-trifluoromethyl-pyridine-2, 4-diol
2, 4-dihydroxy-6-trifluoromethyl-nicotinic acid ethyl ester (62g/247mmol) was added in portions over 30 min to 6M HCl (aq) (620ml) at reflux temperature. The resulting mixture was then heated at 100 ℃ overnight with vigorous stirring to obtain a complete solution. The solution was then cooled and concentrated in vacuo to a white solid. This was slurried in water (250ml) and adjusted to pH7 with concentrated ammonia to give a thick white suspension. The resulting solid was collected by filtration, rinsed with fresh water and dried to give the title compound as a white solid (44.0 g).
1H NMR(d6-DMSO，400MHz)δ 6.05(s，1H)，6.6(s，1H)
Preparation 21
Ethyl- [2, 3-diamino-6- (trifluoromethyl) -pyridin-4-yl ] -benzylcarbamate
Crude ethyl- [ 2-amino-3-nitro-6- (trifluoromethyl) -pyridin-4-yl ] -benzylcarbamate (65gm, 170mmol) was dissolved in ethanol (1000ml) and 10% Pd-C (6gm) was added. The hydrogenation was carried out at 40 ℃ and 40psi for 1 hour to complete the reduction of the nitro group. The catalyst was removed by filtration and the filtrate was evaporated to dryness under reduced pressure to yield a light brown semi-solid. Trituration with tert-butyl methyl ether (150ml), followed by filtration and washing with the same solvent as above (30ml) gave the title compound as a white solid (36gm, 60% yield).
1H NMR(DMSOd6) δ 7.30-7.21(m, 5H), 6.32 (broad s, 1H), 6.15 (broad s, 2H), 5.39 (broad s, 2H), 5.00 (broad d, 1H), 4.25 (broad d), 4.09 (broad d, 2H), 1.12 (broad s, 3H); LRMS (ES)+)m/z 355(MH+)
Preparation 22
Ethyl- [ 2-amino-3-nitro-6- (trifluoromethyl) -pyridin-4-yl ] -benzyl carbamate
Reacting ethyl- [ 2-chloro-3-nitro-6- (trifluoromethyl) -pyridin-4-yl]Benzyl carbamate (63gm, 160mmol) was dissolved in tetrahydrofuran (300ml) and 0.880 ammonia solution (100ml) was added thereto to give two phases. This was transferred to a pressure vessel, sealed and heated to 80 ℃ for 2 hours with stirring. The tetrahydrofuran was evaporated and the residue was partitioned between saturated brine and diethyl ether. The organic extracts were dried over sodium sulfate, filtered and evaporated to give a thick yellow slurry (65 gm)) A crude product; LRMS (ES)+)m/z 385(MH+)，(ES-)m/z 383(M-H)。
Preparation 23
Ethyl- [ 2-chloro-3-nitro-6- (trifluoromethyl) -pyridin-4-yl ] -benzyl carbamate
Benzyl- (2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) -amine (57gm, 170mmol) was dissolved in tetrahydrofuran (750ml) and washed with N2Stirring the mixture. The resulting mixture was then cooled to-5 ℃ in an ice/salt bath. A solution of potassium tert-butoxide (21.2gm, 189mmol) in tetrahydrofuran (200ml) was added dropwise over about 30 minutes, maintaining the temperature between-5 ℃ and 0 ℃ to produce a dark red reaction mixture. The resulting mixture was then stirred at this temperature for 15 minutes, and a solution of ethyl chloroformate (21.4gm, 198mmol) in tetrahydrofuran (100ml) was added dropwise, maintaining the temperature below 5 ℃.
The cooling bath was removed and the reaction mixture was allowed to reach ambient temperature over 1 hour to give a light brown turbid solution. The solvent was evaporated and the residue was partitioned between saturated brine (50ml) and tert-butyl methyl ether (300 ml). The organic phase was washed successively with water (50ml), saturated brine (50ml), dried over sodium sulphate, filtered and evaporated to give a brown oil. This oil was dissolved in n-pentane (250ml) and stored overnight at ambient temperature.
The n-pentane solution was decanted from the dark brown tar that had precipitated out. Evaporation of the solvent gave the title compound as a light brown viscous oil (63gm, 91% yield).
1H NMR(CDCl3)δ 7.28-7.10(m，5H)，4.80(s，2H)4.15(q，2H)1.18(t，3H)；LRMS(ES+)m/z 404/406(MH+)。
Preparation 24
Benzyl- (2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) -amine
4-benzylamino-3-nitro-6-trifluoromethyl-pyridin-2-ol (61.7gm, 19)7mmol) was added to phenylphosphonyl dichloride (180ml) and stirred under N2Heat to 100 ℃ overnight in an oil bath under atmosphere. The starting material was dissolved by heating to give a pale yellow solution. The mixture was then quenched onto ice water (600gm ice +100ml water) to yield a light yellow solid. The solid was filtered off and washed appropriately with water. The solid was dissolved in ethyl acetate (600ml) and washed with aqueous sodium bicarbonate (10% w/v) until no more foaming and the pH of the wash water was basic. The organic layer was dried over sodium sulfate, filtered and evaporated to yield a cloudy yellow solid. The solid was then dissolved in diethyl ether. N-hexane was then added thereto until the solution became cloudy. A thick flocculent solid formed within a few minutes, then filtered off, washed with n-hexane and dried to yield the title compound (60.59gm, 92% yield).
1H NMR(CDCl3) δ 7.44-7.30(m, 5H), 7.04(s, 1H), 6.95(s broad, 1H)4.53(d, 2H); LRMS (ES)+)m/z 332(MH+)。
Preparation 25
2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-ylamine
Benzyl- (2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) -amine (3.1g, 9.3mmol) was stirred in 5ml of concentrated sulfuric acid for 0.5 h, and the solution was carefully poured into a beaker with crushed ice. Successive addition of solid K2CO3Until a basic pH was achieved, and the aqueous solution was extracted with 2 × 50ml EtOAc. The combined organics were washed with MgSO4Dried and concentrated in vacuo to yield 2.2g of the title compound as a pale yellow solid.
1H NMR(CDCl3)δ 7.06(s，1H)，5.87(bs，2H)；LRMS(ESCI)m/z 240[M-H]+
Preparation 26
(2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) carbamic acid ethyl ester
(2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-ylamine (2.2g, 9.1mmol) was stirred in 2-MeTHF (20ml) and triethylenetetramine was addedAmine (1.52ml, 10.9 mmol). The solution was cooled to-5 ℃ in an ice bath, ethyl chloroformate (1.04ml, 10.9mmol) was added dropwise, the solution was warmed to ambient temperature and stirred under nitrogen for 16 hours. Add 20ml EtOAc and 10ml H2O and the phases were separated and washed with another 2X 10ml of saturated brine solution. The organic extract was extracted with MgSO4Dried, concentrated in vacuo and pre-adsorbed on a silica column. Elution with heptane EtOAc9:1 gave 2.1g of the title compound as a white solid.
1H NMR(CDCl3)δ 8.79(s，1H)，8.02(bs，1H)，4.35-4.30(t，2H)，1.38-1.35(qt，3H)；LRMS(ESCI)m/z 312[M-H]+
Preparation 27
(2-benzylamino-3-nitro-6-trifluoromethyl-pyridin-4-yl) carbamic acid ethyl ester
Ethyl 2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) carbamate (300mg, 0.96mmol) was dissolved in 10ml tetrahydrofuran, then benzylamine (0.103ml, 0.96mmol), triethylamine (0.194ml, 1.91mmol) were added and the reaction mixture was stirred at 60 ℃ overnight. The solvent was removed and the solid was dissolved in ethyl acetate/water (50ml/30ml) and the organic layer was washed with MgSO4Dried, concentrated and eluted with a gradient of 0% to 10% methanol in ethyl acetate and purified by silica column chromatography to give 323mg of the title compound as a yellow solid.
1H NMR(CDCl3)：δ 10.75(s，1H)，8.89(s，1H)，8.22(s，1H)，7.35(m，5H)，4.82(d，2H)，4.31(q，2H)，1.36(t，3H)；LRMS(APCI)m/z 385[MH]+
Preparation 28
(3-amino-2-benzylamino-6-trifluoromethyl-pyridin-4-yl) carbamic acid Ethyl ester (2-benzylamino-3-nitro-6-trifluoromethyl-pyridin-4-yl) -carbamic acid Ethyl ester (95mg, 0.25mmol) was dissolved in 10mL ethanol, Raney Nickel (20mg, 20% MW) was added, and the reaction mixture was stirred under 50PSI hydrogen in a pressure pot at room temperatureStirring for 2 hours. The mixture was filtered with Arbocel (lignocelluloses) and the filtrate was concentrated in vacuo to give 88mg of the title compound as a pale green colloid; LRMS (APCI) m/z 355[ MH]+
Preparation 29
4-benzylamino-6-trifluoromethyl-1, 3-dihydro-imidazo (4, 5-c) pyridin-2-one
(3-amino-2-benzylamino-6-trifluoromethyl-pyridin-4-yl) -carbamic acid ethyl ester (88mg, 0.25mmol) was dissolved in 5ml acetic acid and the reaction mixture was stirred at 80 ℃ overnight. The solvent was removed and the gum was dissolved in water/ethyl acetate. The organic layer was separated over MgSO4Dry, remove the solvent in vacuo, and purify by silica column chromatography eluting with a gradient of 1% to 5% methanol in ethyl acetate to yield 37mg of the title compound as a colorless colloid.
1H NMR(CDCl3)：δ 10.56(s，1H)，7.51-7.47(m，5H)，6.61-6.58(m，2H)，5.87(s，1H)，4.61(d，2H)；LRMS(APCI)m/z 309[MH]+
Preparation 30
4-benzylamino-1- (6-methyl-pyridin-3-ylmethyl) -6-trifluoromethyl-1, 3-dihydro-imidazo (4, 5-c) pyridin-2-one
4-benzylamino-6-trifluoromethyl-1, 3-dihydro-imidazo (4, 5-c) pyridin-2-one (100mg, 0.32mmol) was dissolved in 5mL dimethylformamide followed by the addition of potassium carbonate (89mg, 0.65mmol), 5-chloromethyl-2-methyl-pyridine (46mg, 0.32mmol), and the reaction mixture was stirred at 80 ℃ overnight. Mass spectrometry analysis showed some expected benzhydrylation product. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate/water. The organic layer was separated over MgSO4Drying, the solvent was removed in vacuo, and the residue was purified by silica column chromatography eluting with a gradient of 1% to 10% methanol in ethyl acetate to yield 30mg of the title compound as a white solid.
1H NMR(d6 DMSO)：δ 8.46(s，1H)，7.57-7.55(dd，1H)，7.38-7.16(m，7H)，6.63(t，1H)，5.02(s，2H)，4.59(d，2H)，2.40(s，3H)；LRMS(APCI)m/z 414[MH]+
Preparation 31
(2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
Potassium carbonate (88mg, 0.64mmol) was added to a stirred solution of (2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) -carbamic acid ethyl ester (100mg, 0.32mmol) in acetone (10 ml). Then 5- (chloromethyl) -2-methylpyridine (54.2mg, 0.38mmol), sodium iodide (57.4mg, 0.38mmol) were added and the suspension was stirred under nitrogen for 16 h. Add 20ml EtOAc and wash the organic phase with 2 XH2O washing with MgSO 24Dried and concentrated in vacuo to yield a red oil. The crude material was purified by silica column chromatography eluting with heptane, EtOAc 3:2 to yield 54mg of the title compound as an orange solid.
1H NMR(CDCl3)δ 8.35(d，1H)，7.54-7.51(dd，1H)，7.31(s，1H)，7.16-7.15(d，1H)，4.84(s，2H)，4.23-4.18(qt，2H)，2.55(s，3H)，1.26-1.22(t，3H)；LRMS(ESCI)m/z 419[MH]+
Preparation 32
(2-amino-3-nitro-6-trifluoromethyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
(2-chloro-3-nitro-6-trifluoromethyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (54mg, 0.13mmol) was dissolved in THF (1ml) and transferred to a 10ml reaction flask. 880 ammonia (1ml) was added, the vessel sealed and the mixture stirred vigorously at ambient temperature for 16 h. The solution was concentrated in vacuo to give a crude oil which was directly purified by silica column chromatography eluting with 100% EtOAc to give 27mg of the title compound as a yellow residue.
1H NMR(CDCl3)δ 8.39(s，1H)，7.61-7.59(d，1H)，7.15-7.13(d，1H)，6.76(s，1H)，6.25(bs，2H)，4.92(s，2H)，4.23-4.14(qt，2H)，2.54(s，3H)，1.22-1.19(t，3H)：LRMS(APCI)m/z 400[MH]+
Preparation 33
4-benzylamino-3-nitro-6-trifluoromethyl-pyridin-2-ol
4-chloro-3-nitro-6-trifluoromethyl-pyridin-2-ol (65.1gm, 268mmol) was dissolved in tetrahydrofuran (350ml) and at room temperature under N2Stirring under an atmosphere. Benzylamine (86.3gm, 805mmol) in tetrahydrofuran (50ml) was added dropwise over 30 minutes to give a bright yellow solution. The reaction was heated in an oil bath at 50 ℃ for 18 hours (solids formed during the reaction). The resulting mixture was then cooled to ambient temperature, diluted with diethyl ether (200ml) and the resulting solid (benzylamine hydrochloride) filtered off. The filtrate was evaporated to low volume under reduced pressure to give a yellow thick slurry. Diethyl ether (300ml) was added and the yellow solid was filtered off and dried on a filter pad to give benzylamine salt (96.5 gm). The desired product was liberated by dissolving the solid between 2N aqueous HCl and dichloromethane and crystallized from ethyl acetate/N-pentane to yield the title compound as a pale yellow solid (61.7gm, 73.4% yield).
1H NMR(DMSO d6δ 9.04 (broad singlet, 1H), 7.38-7.25(m, 5H), 6.54(s, 1H), 4.67(d, 2H); LRMS (ES)+)m/z 314(MH+)。
Preparation 34
4-chloro-3-nitro-6-trifluoromethyl-pyridin-2-ol
3-Nitro-6-trifluoromethyl-pyridine-2, 4-diol (5.8gm, 26mmol) was heated in phenylphosphonyl dichloride (30ml) at 100 ℃ for 19 hours. The resulting mixture was then cooled and poured onto ice (60gm) and then extracted with ethyl acetate (3 × 50 ml). The combined organic extracts were washed with aqueous sodium bicarbonate (10% w/v) until the washings were basic (pH 8). The dark yellow organic layer was then washed with saturated brine, dried over sodium sulfate, filtered and evaporated to give a yellow colloid. The colloid was triturated with dichloromethane to give a yellow solid which was filtered off and dried (4.65 gm). The solid was dissolved in water (25ml) and acidified with 2N hydrochloric acid (7.5ml) to give a white thick precipitate which was filtered off and washed with water. The precipitate was dissolved in ethyl acetate, dried over sodium sulfate, filtered and evaporated to give the title compound as a white solid (3.75 gm).
1H NMR(DMSOd6)δ 7.78(s，1H)。13C NMR(DMSOd6)δ 157.2(s)145.2(q)138.1(s)136.98(s)120.6(q)113.86(s)；LRMS(ES-)m/z 241/243[MH]-
Preparation 35
3-nitro-6-trifluoromethyl-pyridine-2, 4-diol
6-trifluoromethyl-pyridine-2, 4-diol (56gm, 310mmol) was added in 3-5mg portions to concentrated sulfuric acid (140ml) with stirring to give a light brown solution. The temperature rose to-50 ℃ during the addition. Nitric acid (21.1ml, 328mmol, 70% HNO3, d 1.4gm/ml) was added dropwise over about 90 minutes at a flow rate such that the reaction temperature was maintained between 45 ℃ and 50 ℃. Once all of the nitric acid was added, the reaction was cooled to ambient temperature over a period of 3 hours. The reaction mixture was then poured into ice/water (-1.3 kg) with stirring, after a few minutes a light yellow precipitate formed, which was filtered off, dissolved in ethyl acetate and dried over sodium sulfate, filtered and evaporated. The 2 nd harvest was obtained by extracting the aqueous filtrate with ethyl acetate. The individual extracts were combined and purified by crystallization from ethyl acetate/n-heptane to yield the title compound as a white "fluffy" solid (49.5gm, 71% yield).
1H NMR(DMSOd6)δ 6.82(s，1H)。13C NMR(DMSOd6)159.82(s)157.58(s)143.10 (broad singlet) 127.26(s)120.85(q) 102.83(s).
Preparation 36
2, 6-dibromopyridine 1-oxide
2, 6-dibromopyridine (79g, 334mmol) is dissolved in 800ml of anhydrous dichloromethane and cooled to 5 ℃ under nitrogen atmosphere, then urea hydrogen peroxide (104g, 1.1mol) is added in one portion. When the mixture was cooled again to 3 ℃, a solution of trifluoroacetic anhydride (140ml, 1mol) in 100ml of DCM was added over 45 minutes with a dropping funnel while keeping the temperature between 5-7 ℃. The mixture was warmed to room temperature and stirred for 20 hours. The mixture was cooled to 10 ℃ in an ice bath and 10% Na was added dropwise over 60 minutes 2SO3Aqueous solution (. about.50 g/500ml) until negative by starch iodide paper test. The resulting mixture was filtered to remove a large amount of fluffy solids and the layers were separated. The aqueous layer was extracted with dichloromethane (2X 200ml) and the combined extracts were extracted with MgSO4Dried and concentrated under reduced pressure to give a light brown solid. The crude product was recrystallized using 600ml boiling acetone to yield 48.47g of the title compound.
1H NMR(CDCl3)δ 7.65(d，2H)，6.95(m，1H)。
Preparation 37
2, 6-dibromo-4-nitropyridine 1-oxide
2, 6-dibromopyridine 1-oxide (10g, 39.5mmol) was added to 65ml of concentrated sulfuric acid at room temperature without cooling. Concentrated sulfuric acid (15ml) was mixed with nitric acid (13.3ml) and placed in a pressure-equalizing dropping funnel. The reaction mixture was heated to 79 ℃ and then the nitrating mixture was added in portions over 25 minutes. When the addition was complete, the mixture was stirred at 83-85 ℃ for 3.5 hours. The mixture was cooled to room temperature and poured slowly onto a 250g piece of crushed ice. A very light yellow solid formed which was filtered off and washed with water (100ml) and dried in a vacuum oven at 50 ℃ overnight to give 10.9g of the title compound.
1H NMR(CDCl3)δ 8.45(s，2H)。
Preparation 38
2, 6-dibromopyridin-4-ylamine
2, 6-dibromo-4-nitropyridine 1-oxide (14.5g, 48.6mmol) was dissolved in 130ml of acetic acid, iron powder (11g, 196.9mmol) was added in succession, and the mixture was stirred at room temperature for 45 minutes. 500ml of water was added and the product was extracted with EtOAc (500 ml). The organic layer was washed with 300ml of water and 300ml of saturated K in this order 2CO3The solution was washed with 300ml brine. The organic layer was dried over magnesium sulfate and the solvent was removed in vacuo to yield 11.1g of the title compound as a white solid.
1H NMR(CDCl3) δ 6.65(s, 2H), 4.4-4.1(s broad peak, 2H); LRMS (ES)+)m/z251，253[MH]+
Preparation 39
2, 6-dibromopyridin-4-yl-N-nitroamines
2, 6-dibromopyridin-4-ylamine (11g, 43.6mmol) was dissolved in 100ml of sulfuric acid at room temperature and then cooled at-5 ℃. 6ml of nitric acid was added dropwise, the temperature was kept between-10 ℃ and-5 ℃ and the mixture was stirred at-5 ℃ for 30 minutes. The mixture was then poured onto 400ml of crushed ice. The solid formed was filtered off and then dissolved in EtOAc. The residual water was removed and the organic layer was washed with 300ml brine, dried over magnesium sulfate and the solvent was removed in vacuo to yield 12.5g of the title product as a yellow solid.
1H NMR(CDCl3) δ 6.85(s 1H), 5.7-5.4(s broad peak, 2H).
Preparation 40
2, 6-dibromo-3-nitro-pyridin-4-ylamine
Concentrated sulfuric acid (250ml) was heated in an oil bath until the acid temperature reached 47 ℃. 2, 6-dibromopyridin-4-yl-N-nitroamine (34.0g, 114.5mmol) was added in succession over 35 minutes. The temperature of the mixture gradually rose throughout the addition until it was finally 56 ℃. The mixture was stirred at 53-55 ℃ for 1 hour. Once the reaction was complete, the reaction mixture was cooled in an ice bath with stirring and poured over On 2L crushed ice. The product precipitated and was filtered off. Combined with the other batch 00110916-140-001 obtained from the same scale reaction. The crude wet aminonitropyridine was dissolved in 700ml EtOAc and the aqueous layer was separated. The organic layer was washed with water (2X 150ml), 1X 150ml NaHCO3The aqueous solution, brine (2X 150ml) were washed and dried (MgSO 24) And concentrated under reduced pressure to give 28g of the title compound.
1H NMR(CDCl3) Delta 7.3-7.2(s broad, 1H), 7.55(s, 2H).
Preparation 41
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine
2, 6-dibromo-3-nitro-pyridin-4-ylamine (6.5g, 21.9mmol) was suspended in concentrated HCl (100ml) and cooled to 0 ℃ before sodium nitrite (7.5g, 109mmol) was added. The mixture was stirred for 30 minutes and then warmed to room temperature. 100ml of cold water were added and the mixture was extracted with 100ml of ethyl acetate. The organic layer was washed with 100ml of water, dried over magnesium sulfate, and the solvent was removed in vacuo to give 5.8g of 2, 6-dibromo-3-nitro-4-chloropyridine as an orange oil. This oil was dissolved in 80ml THF and cooled to 0 ℃. Benzylamine (94.9ml, 44.9mmol) was dissolved in 20ml THF and added dropwise to the reaction followed by potassium carbonate (6.6g, 48.2 mmol). The mixture was warmed to room temperature and then heated at 50 ℃ overnight. Once the reaction was complete, the mixture was dissolved in water (150ml) and ethyl acetate (100 ml). The organic layer was washed with 200ml water and 200ml brine, dried over magnesium sulfate and the solvent removed in vacuo. 100ml ethanol was added and the mixture was sonicated for five minutes and placed in a fume hood overnight. The precipitate was filtered off and washed with 30ml of ethanol to give 4.56g of the title compound as a yellow solid.
1H NMR(CDCl3) δ 9.6-9.4(m width peak, 2H), 7.4-7.2(m, 10H), 6.2(s, 1H), 4.8(d, 2H), 4.45(d, 2H); LRMS (ES)+)m/z 413，415[MH]+
Preparation 42
N-2, N-4-dibenzyl-3-nitro-6-vinyl-pyridine-2, 4-diamine
N-2, N-4-dibenzyl-3-nitro-6-vinyl-pyridine-2, 4-diamine (2g, 5mmol) was dissolved in THF (60ml), and vinyltributyltin (3.4g, 10.8mmol), palladium acetate (350mg, 10% by weight), and triphenylphosphine (380mg) were added. The mixture was degassed with argon and then heated at 80 degrees overnight. The solvent was removed in vacuo and the crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane as eluent to yield 1.9g of the title compound as an orange solid.
1H NMR(CDCl3) δ 9.65(s peak width, 1H), 9.4(s peak width, 1H), 7.45-7.2(m, 10H), 6.5-6.4(m, 1H), 6.35(m, 1H), 5.95(s, 1H), 5.5(m, 1H), 4.85(d, 2H), 4.55(d, 2H); LRMS (ES)+)m/z 361[MH]+
Preparation 43
4, 6-bis-benzylamino-5-nitro-pyridine-2-carbaldehyde
N-2, N-4-dibenzyl-3-nitro-6-vinyl-pyridine-2, 4-diamine (800mg, 2mmol) was dissolved in 10ml of tetrahydrofuran and 30ml of water, and osmium tetroxide (60mg, 0.2mmol) and sodium metaperiodate (1.2g, 5.6mmol) were added in this order. The mixture was stirred at room temperature overnight. The mixture was dissolved in water (30ml) and ethyl acetate (30ml), the organic layer was washed with 50ml brine, dried over magnesium sulfate and the crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane as eluent to give 450mg of the title compound as an orange solid.
1H NMR(CDCl3) δ 9.78(s, 1H), 9.55 (broad singlet, 1H), 9..3(s broad peak, 1H), 7.4-7.2(m, 10H), 7.62(s, 1H), 5.5(m, 1H), 4.9(d, 2H), 4.6(d, 2H); LRMS (ES)+)m/z 363[MH]+
Preparation 44
N-2, N-4-dibenzyl-6-morpholin-4-yl-methyl-3-; nitro-pyridine-2, 4-diamines
4, 6-bis-benzylamino-5-nitro-pyridine-2-carbaldehyde (150mg, 0.41mmol) was dissolved in 15ml dichloromethane at room temperature, followed by addition of 2-methoxy-ethylamine (43mg, 0.49mmol), acetic acid (25mg, 0.41 mmol). The mixture was stirred for 5 min, then sodium triacetoxyborohydride (130mg, 0.62mmol) was added and the mixture was stirred at room temperature for 1 h. 20ml of water were added to the mixture, and the organic layer was separated, washed with 20ml of water, and then MgSO4And (5) drying. The solvent was removed in vacuo to yield 180mg of the title compound as an orange colloid.
1H NMR(CDCl3) δ 9.55 (broad singlet, 1H), 9.4 (broad singlet, 1H), 7.4-7.2(m, 10H), 6.19(s, 1H), 4.8(d, 2H), 4.55(d, 2H), 3.6(m, 4H), 3.45(m, 2H), 3.3(s, 2H), 2.35(m, 4H); LRMS (ES)+)m/z 434[MH]+
Preparation 45
N-2, N-4-dibenzyl-6-morpholin-4-yl-methyl-pyridine-2, 3, 4-triamine
N-2, N-4-dibenzyl-6-morpholin-4-yl-methyl-3-nitro-pyridine-2, 4-diamine (190mg, 0.43mmol) was dissolved in 30ml of methanol, Raney nickel (40mg, 20% by weight) was added, and the mixture was stirred at room temperature under 80psi of hydrogen for 1 hour. Upon completion, the mixture was filtered with Arbocel (lignocellulose) and the solvent was removed in vacuo to yield 180mg of the title compound as a green oil.
1H NMR(CD3OD): δ 7.4-7.2(m, 10H), 6.1(s, 1H), 4.6(d, 2H), 4.4(d, 2H), 3.3(m, 6H), 2.4-2.2 (broad singlet, 2H), 2.15(m, 4H); LRMS (ES)+)m/z 417[MH]+
Preparation 46
N-2, N-4-dibenzyl-6-ethyl-pyridine-2, 3, 4-triamine
N-2, N-4-dibenzyl-3-nitro-6-vinyl-pyridine-2, 4-diamine (300mg, 0.75mmol) was dissolved in 20ml of tetrahydrofuran, Raney nickel (40mg, 13% by weight) was added, and the mixture was stirred at room temperature under 60psi of hydrogen for 1.5 hours. Upon completion, the mixture was filtered with Arbocel (lignocellulose) and the solvent was removed in vacuo to yield 230mg of the title compound.
1H NMR(CDCl3): δ 7.40-7.20(m, 5H), 6.0(s, 1H), 4.90-4.80 (broad singlet, 2H), 4.65(d, 2H), 4.35(d, 2H), 2.55(q, 2H), 1.20(t, 3H).
Preparation 47
1- (4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -ethanone
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (800mg, 1.94mmol) was dissolved in 40ml of tetrahydrofuran, followed by the addition of (1-ethoxyvinyl) -tri-N-butyltin (909mg, 2.52mmol), palladium acetate (90mg, W/W), and triphenylphosphine (100mg, W/W) in that order, and the mixture was stirred at 80 ℃ for 1 hour. Once the reaction was complete, 30ml of ethyl acetate and 40ml of 3N HCl solution were added. The mixture was stirred vigorously at 60 ℃ for 30 minutes. The organic layer was separated, washed with 50ml brine and the solvent was removed in vacuo. The crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane to give 680mg of the title compound as a yellow solid.
1H NMR(CDCl3): δ 9.55 (broad singlet, 1H), 9.35 (broad singlet, 1H), 7.40-7.0(m, 10H), 6.75(s, 1H), 5.25(s, 1H), 4.85(d, 2H), 4.55(d, 2H), 2.50(s, 3H); LRMS (ES)+)m/z 377[MH]+
Preparation 48
N-2, N-4-dibenzyl-6-difluoromethyl-3-nitro-pyridine-2, 4-diamine
4, 6-bis-benzylamino-5-nitro-pyridine-2-carbaldehyde (250mg, 0.69mmol) was dissolved in 15ml dichloromethane and cooled to 0 ℃ before bis- (2-methoxyethyl) aminosulfur trifluoride (611mg, 2.76mmol) was added and the reaction mixedThe mixture was stirred at room temperature for 3 hours. Once the reaction was complete, 30ml of water was added to the mixture, and the organic layer was separated, washed with 30ml of saturated potassium carbonate solution and brine, and then MgSO4And (5) drying. The solvent was removed in vacuo and the crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane to yield 220mg of the title compound as a yellow solid.
1H NMR(CDCl3) δ 9.60 (broad singlet, 1H), 9.3 (broad singlet, 1H), 7.4-7.2(m, 10H), 6.30(s, 1H), 6.35-6.05(t, 1H), 6.75(d, 2H), 6.50(d, 2H); LRMS (ES)+)m/z 385[MH]+
Preparation 49
4, 6-bis-benzylamino-5-nitro-pyridine-2-carbonitrile
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (500mg, 1.21mmol) was suspended in 10ml of toluene, and then tributyltin cyanide (765mg, 2.42mmol), palladium acetate (60mg, W/W), and triphenylphosphine (70mg, W/W) were added in this order, and the mixture was microwaved at 130 ℃ for 25 minutes. Once the reaction was complete, the solvent was removed in vacuo and the crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane to yield 408mg of the title compound as a yellow solid.
1H NMR(CDCl3): δ 9.65 (broad singlet, 1H), 9.30 (broad singlet, 1H), 7.40-7.20(m, 10H), 6.40(s, 1H), 4.75(d, 2H), 4.50(d, 2H); LRMS (ES)+)m/z 360[MH]+
Preparation 50
(4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -methanol
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (315mg, 0.87mmol) was suspended in 20ml tetrahydrofuran and cooled to 0 ℃, then sodium boron hydride (40mg, 1.1mmol) was added and the mixture was stirred at 0 ℃ for 15 minutes. The mixture was dissolved in water (10ml) and ethyl acetate (10 ml). The organic layer was separated, washed with 15ml brine, dried over magnesium sulfate and the solvent was removed in vacuo to yield 315mg of the title compound as a yellow solid.
1H NMR(CDCl3): δ 9.50 (broad singlet, 1H), 7.40-7.20(m, 10H), 5.85(s, 1H), 4.59(d, 2H), 4.45(d, 2H), 4.35(s, 2H); LRMS (ES)+)m/z[MH]+
Preparation 51
N-2, N-4-dibenzyl-6-bromomethyl-3-nitro-pyridine-2, 4-diamine
(4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -methanol (300mg, 0.82mmol) was dissolved in 20ml dichloromethane and cooled to 0 ℃ before triphenylphosphine (237mg, 0.91mmol), N-bromosuccinimide (161mg, 0.82mmol) were added in that order and the mixture was stirred at 0 ℃ for 30 minutes, then warmed to room temperature and stirred for 2 hours. The solvent was removed in vacuo and the crude residue was purified by silica gel column chromatography using 20% ethyl acetate in pentane to yield 220mg of the title compound as a yellow solid.
1H NMR(CDCl3): δ 9.59 (broad singlet, 1H), 9.40 (broad singlet, 1H), 7.40-7.20(m, 10H), 6.15(s, 1H), 4.80(d, 2H), 4.55(d, 2H), 4.15(s, 2H); LRMS (ES)+)m/z 427，429[MH]+
Preparation 52
N-2, N-4-dibenzyl-6-methoxymethyl-3-nitro-pyridine-2, 4-diamine
N-2, N-4-dibenzyl-6-bromomethyl-3-nitro-pyridine-2, 4-diamine (100mg, 0.23mmol) was dissolved in 10ml of methanol, followed by addition of sodium methoxide (25mg, 0.46mmol), followed by stirring at 60 ℃ overnight. The solvent was removed in vacuo and the crude product was dissolved in 10ml dichloromethane and 10ml water. The organic layer was separated, dried over magnesium sulfate and the solvent was removed in vacuo to yield 80mg of the title compound as a yellow solid.
1H NMR(CDCl3): δ 9.59 (broad singlet, 1H), 9.40 (broad singlet, 1H), 7.40-7.20(m, 10H), 6.20(s, 1H), 4.80(d, 2H), 4.55(d, 2H), 4.25(s, 2H), 3.35(s, 3H); LRMS (ES)+)m/z 379[MH]+
Preparation 53
N-2, N-4-dibenzyl-3-nitro-6-pyrazin-2-yl-pyridine-2, 4-diamine
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (100mg/0.242mmol), 2-tri-N-butylstannylpyrazine (116mg/0.315mmol), palladium acetate (15mg) and triphenylphosphine (20mg) were dissolved in toluene (2ml) and irradiated with microwaves at 130 ℃ for 25 minutes in a biotageinitator. The reaction was repeated on the same scale in more than 2 occasions. The three reactions were combined, diluted with EtOAc (10mL), washed with water (5mL) and concentrated in vacuo. Purification by column chromatography eluting with 10:1 pentane: EtOAc gave the title compound as a yellow solid (105 mg).
1H NMR (CDCl3, 400MHz) delta 4.60-4.65(d, 2H), 4.90-4.95(d, 2H), 7.20-7.40 (multiplet, 10H), 8.55-8.60 (multiplet, 2H), 9.40-9.50 (multiplet, 2H), 9.60 (multiplet, 1H); LRMS (ESCI) m/z 413[ MH]+。
Preparation 54
N-2, N-4-dibenzyl-6-pyrazin-2-yl-pyridine-2, 3, 4-triamine
N-2, N-4-dibenzyl-3-nitro-6-pyrazin-2-yl-pyridine-2, 4-diamine (105mg/0.255mmol) was dissolved in MeOH (20ml)/THF (20 ml). Raney nickel (30mg) was added and the reaction was allowed to stand at room temperature under 80psi of hydrogen for 5 hours. Filtered through celite and concentrated in vacuo to give the title compound as a pale green oil (95 mg); LRMS (ESCI) m/z 383[ MH]+，381[MH]-。
Preparation 55
6-allyl-N2, N4-dibenzyl-3-nitro-pyridine-2, 4-diamine
Will N*2*，N*4*-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (1g, 2.4mmol) was stirred in anhydrous tetrahydrofuran (20ml) and the solution degassed with nitrogen, followed by the addition of palladium acetate (109mg, 0.48mmol) and allyl tributyltin (1.1ml, 3.6 mmol). The reaction mixture was degassed for 10 minutes and the suspension was heated at 80 ℃ for 16 hours. The suspension was cooled to ambient temperature, concentrated in vacuo and eluted with pentane: EtOAc 9:1 directly purified by silica column chromatography to give the title compound as a yellow solid (696mg, 77%).
1H NMR(CDCl3)δ 3.23-3.25(d，2H)，4.47-4.49(d，2H)，4.81-4.82(d，2H)，5.07-5.14(m，2H)，5.89-5.99(m，1H)，7.29-7.40(m，11H)；LRMS(ES)m/z 375[MH]+
Preparation 56
(4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -acetaldehyde
6-allyl-N2, N4-dibenzyl-3-nitro-pyridine-2, 4-diamine (1.0g, 2.7mmol) was suspended in a mixture of 15ml tetrahydrofuran and 30ml water. After addition of potassium osmate (148mg, 0.4mmol) and sodium periodate (1.17g, 5.4mmol), the solution was stirred vigorously at room temperature for 30 min. Ethyl acetate (20ml) was added to the reaction mixture and the phases were dissolved separately, the organic extract was dried over magnesium sulfate, concentrated in vacuo and eluted with 100% EtOAc, purified by silica column chromatography to give 3- (4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -propane-1, 2-diol (992mg, 91%). The intermediate diol was stirred in 20ml of acetone in the presence of sodium periodate (1.17g, 5.5 mmol). After 2h, the solution was partitioned between EtOAc and water and the organic extract was dried (over MgSO4) And concentrated to give the title compound as an orange oil (962mg, 96%).
1H NMR(CDCl3)δ 3.49(d，2H)，4.49-4.50(d，2H)，4.65-4.66(d，2H)，7.52-7.90(m，11H)，10.03(s，1H)；
LCMS(APCI+)RT@3.74min，m/z 409[MH]+
Preparation 57
2- (4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -ethanol
(4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -acetaldehyde (250mg, 0.66mmol) was stirred in 15ml dichloromethane. Sodium borohydride (38mg, 0.99mmol) was added and the solution was stirred at ambient temperature overnight. The reaction mixture was partitioned between DCM and water, the organics dried over magnesium sulfate and concentrated to give a crude solid, which was purified by silica column chromatography eluting with pentane: EtOAc 4:1-1:1 to give the title compound as a yellow solid (134mg, 53%).
1H NMR(CDCl3)δ 2.68-2.70(t，2H)，3.83-3.86(t，2H)，4.49-4.51(d，2H)，4.72-4.74(d，2H)，5.86(s，1H)，7.29-7.38(m，10H)，9.50-9.54(bd，2H)；LRMS(ES)m/z 379[MH]+
Preparation 58
N2, N4-dibenzyl-6- (2-methoxy-ethyl) -3-nitro-pyridine-2, 4-diamine
2- (4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -ethanol (134mg, 35mmol) was suspended in a mixture of dichloromethane (15ml) and triethylamine (59. mu.l, 0.43mmol) and the solution was cooled to 5 ℃ in an ice bath. Methanesulfonyl chloride (33 μ 1, 0.43mmol) was added and the reaction mixture was stirred at ambient temperature for 1 hour. 10ml DCM were added and the organics were washed with 2 XK2CO3Washed (10% aqueous), dried and concentrated to give a crude oil. The intermediate mesylate salt was suspended in acetone (20ml), sodium methoxide (96mg, 1.7mmol) was added, and the mixture was heated at reflux for 1 hour. The residual solvent was removed in vacuo, DCM was added and the solution was washed with 2 × H2And O washing. The combined organics were washed with MgSO4Dried, concentrated and eluted with 8:1-4:1 Pent: EtOAc, purified by silica column chromatography to give the title compound as a yellow oil (68mg, 47%).
1H NMR(CDCl3)δ 2.70-2.73(t，2H)，3.26(s，3H)，3.64-3.67(t，2H)，4.49-4.50(d，2H)，4.81-4.82(d，2H)，5.91(s，1H)，7.29-7.39(m，10H)，9.40-9.48(bd，2H)；LRMS(ES)m/z 393[MH]+
Preparation 59
N*2*，N*4*-dibenzyl-6- (2-methoxy-ethyl) -pyridine-2, 3, 3-triamine in the presence of Raney nickel (20% by weight, 13mg)*2*，N*4*-dibenzyl-6- (2-methoxy-ethyl) -3-nitro-pyridine-2, 4-diamine (65mg, 0.17mmol) was stirred in THF (10 ml). The mixture was heated at room temperature at 60psi H 2Stir for 2 hours then filter through a pad of Arbocel, washing with 2 × THF. The filtrate was concentrated in vacuo to give the title compound as a brown oil (47mg, 78%) which was used directly in the next step without further purification; LRMS (ES) m/z363[ MH]+
Preparation 60
1-benzyl-4-benzylamino-6- (2-methoxy-ethyl) -1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
Will N*2*，N*4*-dibenzyl-6- (2-methoxy-ethyl) -pyridine-2, 3, 3-triamine (47mg, 0.13mmol) was stirred in acetonitrile (5 ml). N, N-carbonyldiimidazole (105mg, 0.65mmol) was added and the mixture was heated under reflux for 16 hours. The solution was concentrated in vacuo and washed with 100% DCM to 96: 4 elution, direct purification by silica column chromatography to give the title compound as a light brown solid present as a 3:2 mixture with the isomer 1-deazapurine (32mg, 64%); LRMS (ES) m/z 389[ MH)]+
Preparation 61
N2, N4-dibenzyl-6- [2- (2-methoxy-ethylamino) -ethyl ] -3-nitro-pyridine-2, 4-diamine
Will N*2*，N*4*-dibenzyl-3-nitro-6-vinyl-pyridine-2, 4-diamine (50mg, 0.14mmol) was suspended in 2-methoxy-ethylamine (1ml) and the mixture was refluxed for 1 hour. Excess reagent was removed in vacuo and the residue was dissolved in DCM (10ml) and washed with 2 XH 2O washed, dried and concentrated. The crude material was purified by silica column chromatography eluting with DCM: MeOH 92:8 to give the title compound as a yellow oil (58mg, 96%).
1H NMR(CDCl3)δ 2.75-2.80(m，4H)，2.96-3.00(t，2H)，3.29(s，3H)，3.48-3.51(t，2H)，4.48-4.49(d，2H)，4.76-4.78(d，2H)，5.87(s，1H)，7.27-7.40(m，10H)，9.41-9.51(dt，2H)；LRMS(ES)m/z 436[MH]+
Preparation 62
[2- (4, 6-bis-benzylamino-5-nitro-pyridin-2-yl) -ethyl ] - (2-methoxy-ethyl) -carbamic acid tert-butyl ester
Will N*2*，N*4*-dibenzyl-6- [2- (2-methoxy-ethylamino) -ethyl]-3-nitro-pyridine-2, 4-diamine (150mg, 0.34mmol) was suspended in DCM (10ml) and the solution was cooled to 0 ℃ and then boc anhydride (95. mu.l, 0.41mmol) was added dropwise as a solution in 5ml DCM. The mixture was warmed to room temperature and after 1 hour, 10ml of H was used2And (4) quenching O. The organic extracts were dried and concentrated in vacuo to give the title compound as a yellow oil (170mg, 92%).
1H NMR(CDCl3)δ 1.41(s，9H)，2.68(t，2H)，3.30(s，3H)，3.42(bs，2H)，3.50(m，4H)，4.47-4.48(d，2H)，4.80-4.81(d，2H)，5.82(s，1H)，7.24-7.39(m，10H)；LRMS(ES)m/z 536[MH]+
Preparation 63
N2, N4-dibenzyl-3-nitro-6-oxazol-2-yl-pyridine-2, 4-diamine
Butyllithium (12.8ml, 20.5mmol) was added dropwise to a stirred solution of oxazole (1.13ml, 17.1mmol) in anhydrous THF (20ml) at-78 ℃ (dry ice/acetone bath), maintaining a slow rate of addition such that the reaction temperature did not exceed-60 ℃. The solution was stirred at this temperature for 10 minutes, then a solution of zinc chloride (5.00g, 36.7mmol) in THF (30ml) was added dropwise. The solution was stirred at-78 ℃ for 15 minutes, then the cooling bath was removed and the reaction mixture was warmed to room temperature.
An aliquot of the reaction mixture (19ml) was added with a syringe to a pre-sealed and nitrogen purged microwave vial (Biotage, 10-20ml) containing N*2*，N*4*-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (1.11g, 2.68mmol) and bis (triphenylphosphine) palladium dichloride (373mg, 0.53 mmol). The vial was heated under microwave irradiation (Biotage Initiator 8) at 130 ℃ for 15 minutes. The reaction mixture was concentrated in vacuo and then partitioned between 2-methyl THF (80ml) and saturated ammonium chloride solution (80 ml). The mixture was filtered and then transferred to a separatory funnel. The layers were separated and the aqueous solution was extracted with more 2-methyl THF (50 ml). The combined organics were dried (over MgSO4) And is evaporated. The resulting brown solid was triturated with EtOAc and the solid was collected by filtration and then washed with EtOAc to give the product as a brown solid (1.03g, 96%).
1H NMR(CDCl3)δ 4.59(d，J＝5.47Hz，2H)4.91(d，J＝5.47Hz，2H)6.93(s，1H)7.21-7.47(m，11H)7.78(s，1H)9.31-9.44(m，1H)9.54-9.63(m，1H)；LRMS(ES+)m/z 402[MH]+
Preparation 64
N2, N4-dibenzyl-6-oxazol-2-yl-pyridine-2, 3, 4-triamine
Will N*2*，N*4*-dibenzyl-3-nitro-6-oxazol-2-yl-pyridine-2, 4-diamine (1.02g, 2.54mmol) was dissolved in THF (60ml) followed by addition of MeOH (60 ml). The solution was hydrogenated over Raney nickel (210mg, 0.25mmol) under a hydrogen atmosphere (80psi) for 1 h. The reaction mixture was filtered through a pad of celite and then evaporated to give the title compound as a brown colloid (944mg, 100 mg) %). For stability, it was used without further purification.
LCMS Rt＝2.41m/z 372[MH]+
Preparation 65
N2, N4-dibenzyl-6- (1-methyl-1H-imidazol-2-yl) -3-nitro-pyridine-2, 4-diamine
N-methylimidazole (0.728ml, 9.18mmol) was dissolved in anhydrous THF (25ml) and the solution was then cooled to-15 deg.C (ice/salt bath). N-butyllithium in hexane (6.31ml, 10.1mmol) was added dropwise to the solution (color changed from colorless to yellow). The solution was stirred at-15 ℃ for 1 hour, then a solution of anhydrous zinc chloride (5.00g, 36.7mmol) in anhydrous THF (35ml) was added dropwise. The solution was stirred at-15 ℃ for 1 hour, then slowly warmed to room temperature, and then stirred for 1 more hours.
An aliquot (16ml) of this solution was added to a pre-sealed and nitrogen purged microwave bottle (Biotage, 2.0-5.0ml) containing N*2*，N*4*-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (400mg, 0.968mmol) and bis (triphenylphosphine) palladium dichloride (136mg, 0.193 mmol). The vial was heated at 130 ℃ for 15 minutes under microwave irradiation (Biotage Initiator 8). The reaction mixture was concentrated in vacuo. The residue was partitioned between EtOAc (15ml) and 2M ammonia solution (15 ml). Most of the aqueous phase was removed using a separatory funnel (with a small amount of emulsion between the layers). The organic layer was washed successively with more 2M ammonia solution (15ml), brine (15ml) and then dried (over MgSO 4) And is evaporated. The crude product was subjected to column chromatography on an Isco company silica column (12g, Redisep). Elution with EtOAc in heptane increased the gradient linearly from 20:80 to 60:40 over 8 column volumes, then 4 column volumes at 60:40 isocratic. The desired eluting components were combined and evaporated to give the title compound as a yellow solid (240mg, 60%).
1H NMR(CD3OD)δ3.88(s，3H)4.67(d，J＝5.48Hz，2H)4.83(d，J＝5.48Hz，2H)6.92(d，J＝1.17Hz，1H)7.14(d，J＝1.17Hz，1H)7.17(s，1H)7.26-7.42(m，10H)9.43-9.66(m，2H)；LRMS(ES+)m/z 415[MH]+
Preparation 66
N2, N4-dibenzyl-6- (1-methyl-1H-imidazol-2-yl) -pyridine-2, 3, 4-triamine
Will N*2*，N*4*-dibenzyl-6- (1-methyl-1H-imidazol-2-yl) -3-nitro-pyridine-2, 4-diamine (0.235g, 0.567mmol) was dissolved in THF (10ml) followed by addition of MeOH (10 ml). The solution was hydrogenated over Raney nickel (0.050g, 0.58mmol) under a hydrogen atmosphere (80psi) for 1 hour. The reaction mixture was filtered through a pad of celite and then evaporated to give the title compound as a pale green solid (218mg, 100%). For stability, it was used without further purification.
LCMS Rt＝2.22m/z 385[MH]+
Preparation 67
(2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester
A solution of ethyl chloroformate (5.96g) in anhydrous 2-methyl THF (50ml) was added dropwise to a solution of 2, 6-dibromo-3-nitro-pyridin-4-ylamine (15.00g) and triethylamine (10.1g) in anhydrous 2-methyl THF (100ml) at 0 ℃ maintaining the rate of addition so that the reaction temperature did not rise by more than 5 ℃. The reaction mixture was warmed to room temperature and then stirred under nitrogen for 1 hour. An additional portion of ethyl chloroformate (0.54g) was added and the mixture was stirred for an additional 1 hour. Water (50ml) was added and the layers were separated. The aqueous layer was extracted with EtOAc (50mL) and the combined organics were dried (over MgSO 4) And evaporated to a brown solid. This solid was pre-adsorbed on silica (. about.19 g) and then subjected to column chromatography on an Isco company silica column (330g, Redispe) eluting with EtOAc: heptane. The gradient of 1 Column Volume (CV) was made to be 10:90 equal degrees and then increased linearly from 10:90 to 30:70 over 6 CVs. This gave the title compound (12.6g) as a pale yellow foaming solid.
1H NMR (400MHz, chloroform-d) δ ppm 1.36(t, J ═ 7.10Hz, 3H), 4.26(q ═ 7.10Hz, 2H), 7.95(br, s, 1H), 8.59(s, 1H), LCMS Rt＝3.22m/z368，370，372[MH]+
Preparation 68
(2, 6-dibromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
Potassium carbonate (7.95g) was added to a stirred solution of ethyl (2, 6-dibromo-3-nitro-pyridin-4-yl) carbamate (10.62g) in acetone (100 ml). Then, 5- (chloromethyl) -2-methylpyridine (4.89g) and sodium iodide (5.18g) were added in this order. The mixture was stirred under nitrogen for 18 hours. The reaction mixture was filtered, concentrated in vacuo and then partitioned between ethyl acetate (100ml) and water (100 ml). The organics were dried (over MgSO4) And evaporated to a dark purple gel, eluted with EtOAc: heptane, and subjected to column chromatography on an Isco company silica column (330g, Redisep) increasing linearly from 40:60 to 80:20 over a 6 column volume gradient. This gave the title compound (8.5g) as a green gum which solidified upon standing to a pale green solid.
1H NMR (400MHz, chloroform-d) δ ppm 1.24(t, J ═ 7.10Hz, 3H) 2.59(s, 3H) 4.19(q, J ═ 7.10Hz, 2H) 4.79(s, 2H) 7.17(s, 1H) 7.19(d, J ═ 8.20Hz, 1H) 7.57(dd, J ═ 8.20, 2.34Hz, 1H) 8.38(d, J ═ 2.34Hz, 1H), LCMSRt＝2.44m/z 473，475，477[MH]+
Preparation 69
(2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
(2, 6-dibromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (6.00g) was dissolved in 2-methyltetrahydrofuran (60 ml). The solution was aliquoted into 3 sealable containers (Biotage, 10-20 ml). An aqueous ammonia solution (0.88 gcm)-320ml) were added to each bottle (total 60 ml). The bottle is sealed and the bottle is sealed,the biphasic mixture was then stirred vigorously at room temperature overnight. The three reaction mixtures were combined and transferred to a separatory funnel. Ethyl acetate (120ml) and water (120ml) were added. The phases were separated and the organics were washed with brine (100 ml). The organics were dried (over MgSO4) And then evaporated to brown gum. This gum was redissolved in ether and then evaporated to form the title compound as a yellow foaming solid (5.4 g).
1H NMR (400MHz, chloroform-d) δ ppm 1.16-1.27(m, 3H) 2.58(s, 3H) 4.10-4.21(m, 2H) 4.87(s, 2H) 6.34(s, 2H) 6.61(s, 1H) 7.18(d, J ═ 8.19Hz, 1H) 7.66(s, 1H) 8.41(d, J ═ 2.34Hz, 1H), LCMSR (LCMSR), LCMSR (r), c t＝1.94m/z 412[MH]+
Preparation 70
[ 2-amino-6- (4-methyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
Butyllithium (1.6M in hexane, 366 μ l) was added dropwise to a stirred solution of 4-methyloxazole (41mg) in THF (0.5ml) in a reaction flask at-78 ℃ (dry ice/acetone bath). The solution was stirred at this temperature for 10 minutes, then a solution of zinc chloride (199mg) in THF (1ml) was added dropwise. The solution was stirred at-78 ℃ for 15 minutes, then the cooling bath was removed and the reaction mixture was warmed to room temperature. This zinc oxazole solution was added by syringe to a pre-sealed nitrogen purged microwave vial (Biotage, 0.5-2.0ml) containing (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) and bis (triphenylphosphine) palladium dichloride (34 mg). The vial was heated under microwave irradiation (BiotageInitiator 8) at 60 ℃ for 15 minutes. The reaction mixture was then partitioned between ethyl acetate (10ml) and saturated aqueous ammonium chloride (10 ml). The layers were separated and the aqueous solution was extracted with ethyl acetate (10 ml). The combined organics were washed with brine (10ml) and then dried (over MgSO 4) And is evaporated. The crude product was eluted with 4 Column Volumes (CV) of ethyl acetate and subjected to column chromatography on an Isco company silica column (12g, Redisp),the methanol gradient in ethyl acetate was then increased linearly from 0 to 5% over 10 CVs. The title compound (69mg) was thus formed as a yellow gum.
1H NMR (400MHz, chloroform-d) δ ppm 1.11-1.25(m, 3H) 2.28(s, 3H) 2.56(s, 3H) 4.13-4.23(m, 2H) 4.96(s, 2H) 6.37(s, 2H) 7.16(d, J ═ 7.80Hz, 1H) 7.26(s, 1H) 7.53(s, 1H) 7.65-7.78(m, 1H) 8.43(s, 1H), LCMS Rt＝1.86m/z 413[MH]+
Preparation 71
5-ethyl-oxazoles
Ethyl-5-ethyl oxazole-4-carboxylate (3.5g) was dissolved in ethanol (45ml) and a solution of sodium hydroxide (2.07g) in water (18ml) was added. The reaction was stirred at room temperature for 16 hours. The reaction mixture was reduced to-20 ml, then concentrated hydrochloric acid was added to produce a pH of-1-2. Reacting the mixture with CH2Cl23X 30ml extraction. The combined organic extracts were washed with saturated brine and Na2SO4Dried, filtered and evaporated to give a pale yellow solid. This was dissolved in quinoline (3ml), and 100mg of copper (II) oxide was added. The reaction was then heated under slightly reduced pressure (oil bath 160 ℃) and the clear liquid was distilled off at-60-70 ℃. The title compound (790mg) was thus formed as a clear oil.
1H NMR (400MHz, chloroform-d) delta ppm 1.2(t, 3H), 2.6(q, 2H), 6.65(s, 1H), 7.7(s, 1H).
Preparation 72
[ 2-amino-6- (5-ethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 5-ethyl-oxazole (47mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (79 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.13-1.25(m, 3H) 1.33(t, J ═ 7.61Hz, 3H)2.56(s, 3H) 2.80(q, J ═ 7.41Hz, 2H) 4.13-4.22(m, 2H) 4.96(s, 2H)6.31-6.45(m, 2H) 6.97(s, 1H) 7.16(d, J ═ 7.80Hz, 1H) 7.26(s, 1H)7.63-7.79(m, 1H) 8.43(s, 1H), LCMS Rt＝2.16m/z 427[MH]+
Preparation 73
5-isopropyl-oxazole-4-carboxylic acid ethyl ester
Ethylisocyanoacetate (4.52g) was added dropwise to a stirred suspension of KOtBu in THF (35ml) at 0 ℃ under a nitrogen atmosphere. After the addition was complete, the dark brown solution was stirred for 30 minutes and then a solution of isobutyryl chloride (2.1ml) in THF (15ml) was added dropwise, keeping the temperature below about 10 ℃. The reaction was stirred for 1 hour and then evaporated to dryness. The residue was treated with acetic acid (1.14ml) and water (25ml) and then extracted with ether (3X 30 ml). The combined ether extracts were washed with saturated brine, dried over sodium sulfate, filtered and evaporated to give a brown oil which was purified by column chromatography eluting with 1% MeOH in dichloromethane. This gave the title compound (1.91g) as a colorless oil.
1H NMR (400MHz, chloroform-d) delta ppm 1.25(d, 6H), 1.38(t, 3H), 3.8(m, 1H), 4.35(q, 2H), 7.7(s, 1H), LRMS m/z (API)184[ MH]+，367[2MH]+
Preparation 74
5-isopropyl-oxazoles
Ethyl 5-isopropyl-oxazole-4-carboxylate (1.89g) was dissolved in a solution of 1N sodium hydroxide (10ml) and ethanol (0.5ml), and the mixture was stirred at room temperature for 16 hours. 1N HCl solution (ca. 9ml) was added and the mixture was stirred for a few minutes. A white solid crystallized out and was collected by filtration. After drying, this solid was dissolved in quinoline (3ml), and copper oxide (120mg) was added. The reaction was heated under vacuum to ramp the oil bath temperature to-170 ℃. The clear liquid was distilled off to yield a mixture of the desired product and quinoline. This oil was then redistilled at lower pressure (. about.180 mBar) and temperature (70 ℃) to give the title compound as a clear oil (260 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.24(d, 6H), 2.96(m, 1H), 6.7(s, 1H), 7.7(s, 1H)
Preparation 75
[ 2-amino-6- (5-isopropyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 5-isopropyl-oxazole (54mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (48 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.13-1.26(m, 3H) 1.34(d, J ═ 6.63Hz, 6H)2.56(s, 3H) 3.04-3.15(m, 1H) 4.10-4.23(m, 2H) 4.96(s, 2H) 6.39(s, 2H) 6.94(s, 1H) 7.16(d, J ═ 7.80Hz, 1H) 7.21(s, 1H) 7.63-7.76(m, 1H) 8.43(s, 1H), LCMS R1t＝2.30m/z 441[MH]+
Preparation 76
[ 2-amino-6- (4, 5-dimethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4, 5-dimethyl-oxazole (47mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (75 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.03-1.26(m, 3H) 2.19(s, 3H) 2.37(s, 3H) 2.56(s, 3H) 4.09-4.26(m, 2H) 4.95(s, 2H) 6.40(s, 2H)7.16(d, J ═ 7.80Hz, 1H) 7.21(s, 1H) 7.62-7.83(m, 1H) 8.42(s, 1H), LCMS Rt＝2.12m/z 427[MH]+
Preparation 77
Oxazol-4-yl-carbinols
DIBAL-H (56ml of a 1.0M solution in toluene) was added dropwise to a solution of oxazole-4-carboxylic acid ethyl ester (7.50g, 53.1mmol) in THF (140ml) at-78 ℃ over 15 minutes. The resulting solution was stirred at-78 ℃ for 30 min and DIBAL-H (56ml of a 1.0M solution in toluene, 56.0mmol) was added over 15 min. The reaction was then slowly warmed from-78 ℃ to room temperature over a 16 hour period. The resulting bright yellow solution was cooled to 0 ℃ in an ice bath and Na was added in small portions 2SO4.10H2O (15.9 g-equivalent to the weight of DIBAL-H added) (note: slow addition to prevent exotherm) to promote precipitation of the aluminum salt. The mixture was warmed to room temperature and after stirring for 90 minutes, the resulting suspension was filtered through a layer of celite. The celite plug was rinsed with dichloromethane (3 × 100ml) and methanol (2 × 100ml) and the filtrates combined. The solvent was removed under reduced pressure to give the title compound as a brown oil (4.8 g).
1H NMR (400MHz, chloroform-d) delta ppm 4.60(s, 2H), 7.6(s, 1H), 7.9(s, 1H)
Preparation 78
4-methoxymethyl-oxazoles
Oxazol-4-yl-methanol (750mg) was dissolved in anhydrous THF (38ml) and the solution was cooled to 0 ℃. Sodium hydride (365mg, 9.1mmol) was then added in small portions over 4 minutes and, after the addition was complete, the reaction was warmed to room temperature for 30 minutes. The reaction was cooled again to 0 ℃ and the methyl tosylate (2.11g) was added in small portions. After the addition was complete, the reaction was warmed to room temperature and stirred for 16 hours. The crude reaction mixture was pre-adsorbed on silica gel and then subjected to ISCOMBI-flash chromatography (SiO)2(ii) a Gradient elution of 2 to 5% MeOH in DCM, 1% NH3) Purify to give the title compound as a pale yellow liquid (473 mg).
1H NMR (400MHz, chloroform-d) delta ppm 3.41(s, 3H), 4.40 (g: (g)) s，2 H)，7.61(s，1H)，7.85(s，1 H)
Preparation 79
[ 2-amino-6- (4-methoxymethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-methoxymethyl-oxazole (110mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (200mg) and bis (triphenylphosphine) palladium dichloride (68mg) to give the product as a yellow gum (107 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.11-1.29(m, 3H) 2.54(s, 3H) 3.47(s, 3H) 4.09-4.24(m, 2H) 4.49(s, 2H) 4.96(s, 2H) 6.34(s, 2H)7.13(d, J ═ 7.81Hz, 1H) 7.32(s, 1H) 7.63-7.72(m, 1H) 7.76(s, 1H)8.42(s, 1H), LCMS Rt＝1.62m/z 443[MH]+
Preparation 80
(2-amino-3-nitro-6-thiazol-2-yl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70, using thiazole (42mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (79 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.06-1.30(m, 3H) 2.56(s, 3H) 4.07-4.25(m, 2H) 4.99(s, 2H) 6.26(s, 2H) 7.16(d, J ═ 7.80Hz, 1H) 7.46(s, 1H) 7.54(d, J ═ 3.12Hz, 1H) 7.64-7.84(m, 1H) 7.96(d, J ═ 3.12Hz, 1H) 8.35-8.50(m, 1H), LCMS R1 t＝2.11m/z 415[MH]+
Preparation 81
2-chloro-3-oxo-pentanoic acid ethyl ester
Sulfuryl chloride (6.50ml) was added dropwise to propionyl ethyl acetate (11.70g) at room temperature and the reaction was stirred for 16 hours. The reaction mixture was placed under vacuum for 1 hour to remove highly volatile materials, then the residue was distilled under "high" vacuum to give an oil which was distilled at 75-79 ℃ under 6Mbar (═ 4.5mmHg) vacuum to give the title compound as a clear oil (13.54 g).
1H NMR (400MHz, chloroform-d) delta ppm 1.11(t, 3H), 1.31(t, 3H), 2.75(q, 2H), 4.25(q, 2H), 4.80(s, 1H)
Preparation 82
4-Ethyl-oxazole-5-carboxylic acid ethyl ester
2-chloro-3-oxo-pentanoic acid ethyl ester (13.5g) was dissolved in 75ml of 95% formic acid. Ammonium formate (27.6g) was added and the reaction was heated at reflux temperature under nitrogen atmosphere for 6 hours. After cooling to room temperature, the reaction mixture was evaporated and the residue was extracted with ether (3X 50 ml). The combined ether extracts were washed with water and brine, dried (MgSO4) Filtered and evaporated to give a crude oil (9 g). Using oil with CH2Cl2Purification by silica gel chromatography eluting with/MeOH 99:1 gave the title compound as a light brown oil (3.77 g).
1H NMR (400MHz, chloroform-d) delta ppm 1.25(t, 3H), 1.41(t, 3H), 2.90(q, 2H), 4.41(q, 2H), 7.85(s, 1H)
Preparation 83
4-ethyl-oxazole-5-carboxylic acid
4-Ethyl-oxazole-5-carboxylic acid ethyl ester (4.6g) was placed in a solution of 1N NaOH (25ml) and ethanol (1ml) and stirred. The reaction mixture was then stirred at room temperature for 16 hours. Diethyl ether (25ml) was added and the aqueous layer was separated and acidified with 1N HCl (26 ml). The yellow solid formed was collected by filtration and then washed with water, n-pentane to give the title compound as a white solid (2.6 g).
1H NMR (400MHz, chloroform-d) delta ppm 1.30(t，3 H)，2.95(q，2 H)，8.0(s，1H)
Preparation 84
4-ethyl-oxazoles
4-Ethyl-oxazole-5-carboxylic acid (1.2g) was dissolved in quinoline (3ml), and CuO (50mg) was added. The reaction was then heated to 215-20 ℃ and the colorless fraction was collected, yielding the title compound (621mg) as a cloudy oil.
1H NMR (400MHz, chloroform-d) delta ppm 1.21(t, 3H), 2.55(q, 2H), 7.4(s, 1H), 7.8(s, 1H)
Preparation 85
[ 2-amino-6- (4-ethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-ethyl-oxazole (47mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (74 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.09-1.22(m, 3H) 1.25(t, J ═ 7.42Hz, 3H)2.50(s, 3H) 2.56-2.68(m, 2H) 4.04-4.21(m, 2H) 4.92(s, 2H) 6.33(s, 2H) 7.10(d, J ═ 7.81Hz, 1H) 7.23(s, 1H) 7.46-7.49(m, 1H) 7.59-7.69(m, 1H) 8.39(s, 1H), LCMS Rt＝2.07m/z 427[MH]+
Preparation 86
1-bromo-3-methyl-butan-2-one
A solution of 3-methyl-2-butanone (5g) in methanol (55ml) was cooled to-30 ℃. Bromine (2.97 μ l) was then added dropwise and once the addition was complete, the reaction was warmed to room temperature and stirred for 3.5 hours. The reaction was then poured into water (100ml) and extracted with diethyl ether (2X 100ml) and the combined extracts dried (over MgSO4) Filtered and concentrated. The oily residue forms two layers after standingSeparating the same oil layer. The bottom layer was retained to give the title compound (5.14g) as a pale yellow oil.
1H NMR (400MHz, chloroform-d) delta ppm 1.15(d, 6H), 2.95(m, 1H), 3.95(s, 1H)
Preparation 87
4-isopropyl-oxazoles
1-bromo-3-methyl-butan-2-one (1.0g) was added to formamide (3.0ml) and the reaction mixture was heated to 110 ℃ for 6 hours. After cooling to room temperature, the reaction mixture was diluted with 40% potassium hydroxide solution (10ml), stirred for a few minutes and then extracted with ether (3X 10 ml). The ether extracts were combined and carefully evaporated. The resulting brown motor oil was triturated with n-pentane and the solvent decanted. This procedure was repeated (. times.2) and the residual pentane was evaporated to give the title compound as a light brown oil (65 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.30(d, 6H), 2.85(m, 1H), 4.41(q, 2H), 7.35(s, 1H), 7.80(s, 1H)
Preparation 88
[ 2-amino-6- (4-isopropyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-isopropyl-oxazole (54mg) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) to give the product as a yellow gum (71 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.10-1.22(m, 3H) 1.27(d, J ═ 6.64Hz, 6H)2.50(s, 3H) 2.82-2.97(m, 1H) 3.96-4.27(m, 2H) 4.92(s, 2H) 6.34(s, 2H) 7.10(d, J ═ 7.81Hz, 1H) 7.23(s, 1H) 7.45(s, 1H) 7.59-7.70(m, 1H) 8.39(s, 1H), LCMS R1t＝2.23m/z 441[MH]+
Preparation 89
Benzyl- (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester
To a stirred solution of (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (7.44g) in acetone (100ml) was added potassium carbonate (5.57g), followed by benzyl bromide (2.87ml) and sodium iodide (3.63g) in that order. The mixture was stirred under nitrogen for 36 hours. The reaction mixture was then filtered to remove precipitated white solid, concentrated in vacuo, and then partitioned between ethyl acetate (100ml) and water (100 ml). The organics were dried (over MgSO 4) And evaporated to a yellow oil. This crude oil was adsorbed on silica gel and then purified by chromatography on an Isco company silica column (80g, Redisep) eluting with EtOAc in a gradient increasing linearly from 10:90 to 50: 50. This gave the title compound (7.50g) as a yellow oil.
1H NMR (400MHz, chloroform-d) delta ppm 1.20(t, 3H), 4.18(q, 2H), 4.8(br, s, 2H), 7.0(s, 1H), 7.20(m, 2H), 7.35(m, 3H), LRMS m/z (API)458, 460, 462[ MH ]]+
Preparation 90
(2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
Benzyl- (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (7.50g) was dissolved in 2-methyl-tetrahydrofuran (15ml) and the solution was placed in a sealable container. Concentrated aqueous ammonia (15ml) was added and the bottle was sealed and the biphasic mixture was stirred vigorously at room temperature for 36 hours. The reaction mixture was then transferred to a separatory funnel, and ethyl acetate (120ml) and water (120ml) were added. The phases were separated and the organics were washed with brine (100ml) and dried (over MgSO4) And evaporated to a yellow oil. After standing, a yellow solid crystallized out. This was collected by filtration and washed with pentane to give the title compound as a yellow solid (6.64 g).
1H NMR (400MHz, chloroform-d) delta ppm1.21(t, 3H), 4.19(q, 2H), 4.82(br，s，2H)，6.25(br，s，2H)，6.55(s，1H)，7.20-7.35(m，5H)，LRMS m/z(API)395，397[MH]+
Preparation 91
[ 2-amino-6- (4-methoxymethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-methoxymethyl-oxazole (114mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (200mg) and bis (triphenylphosphine) palladium dichloride (71mg) to give the product as a yellow gum (160 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.20(t, J ═ 7.03Hz, 3H) 3.46(s, 3H) 4.12-4.22(m, 2H) 4.47(s, 2H) 4.92-5.03(m, 2H) 6.32(s, 2H) 7.28-7.35(m, 6H) 7.74(s, 1H), LCMS Rt＝3.08m/z 428[MH]+
Preparation 92
Dimethyl-oxazol-4-ylmethyl-amine
Thionyl chloride (5.51ml) was added dropwise over 5 minutes to a cold solution (0 ℃) of oxazol-4-yl-methanol (1.51g) in dichloromethane (50 ml). The resulting cloudy solution was stirred at room temperature for 5 minutes and then heated to reflux. Upon heating, the solution became clear and dark yellow. After 10 minutes at reflux temperature, the solution was cooled to room temperature and the excess thionyl chloride and solvent were removed under reduced pressure to give the corresponding chloride compound, which was used without further purification. Dimethylamine (38ml of a 2.0M solution in THF) was cooled to 0 ℃ in an ice bath and a solution of the chloride (1.74g) in anhydrous THF (50ml) was added in small portions over 10 minutes. The resulting suspension was then allowed to react for 16 hours. The solvent was removed to give a dark brown/black solid which was then pre-adsorbed on silica and purified by ISCO combi-flash chromatography on silica gel eluting with 5 to 15% MeOH in dichloromethane with 10% NH 3. This gave the title compound (335mg) as a dark brown viscous oil.
1H NMR (400MHz, chloroform-d) delta ppm 2.35(s, 6H), 3.50(s, 2H), 7.62(s, 1H), 7.85(s, 1H)
Preparation 93
[ 2-amino-6- (4-dimethylaminomethyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using dimethyl-oxazol-4-ylmethyl-amine (128mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (200mg) and bis (triphenylphosphine) palladium dichloride (71mg) to give the product as a yellow gum (99 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.20(t, J ═ 7.03Hz, 3H) 2.32(s, 6H) 3.49(s, 2H) 4.13-4.23(m, 2H) 4.92-5.03(m, 2H) 6.33(s, 2H) 7.28-7.36(m, 6H) 7.68(s, 1H), LCMS Rt＝1.86m/z 441[MH]+
Preparation 94
{ 2-amino-3-nitro-6- [1- (2-trimethylsilyl-ethoxymethyl) -1H-imidazol-2-yl ] -pyridin-4-yl } -benzyl-carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 1- (2-trimethylsilyl-ethoxymethyl) -1H-imidazole (201mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (200mg) and bis (triphenylphosphine) palladium dichloride (71mg) to give the product as a yellow gum (174 mg).
1H NMR (400MHz, chloroform-d) δ ppm-0.04(s, 9H) 0.87-0.94(m, 2H) 1.19(t, J ═ 7.03Hz, 3H) 3.49-3.59(m, 2H) 4.10-4.22(m, 2H) 4.98(s, 2H) 5.93(s, 2H) 6.18(s, 2H) 7.18(s, 1H) 7.23(s, 1H) 7.28-7.38(m, 5H) 7.53(s, 1H)
Preparation 95
(2-amino-6-methylsilyl-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
Sodium thiomethoxide (180mg) was added dropwise to a stirred suspension of (2-amino-6-chloro-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (300mg) in methanol: THF (3:1, 4 ml). The mixture was sealed in a reaction flask and then stirred at room temperature for 2 hours. The reaction mixture was diluted with methanol and then directly pre-adsorbed onto silica gel. The crude material was column chromatographed using EtOAc: heptane, through an Isco company silica column (12g, Redispe) with a 6 column volume gradient increasing linearly from 30:70 to 50: 50. The desired eluted fractions were combined and evaporated to give the title compound (297mg) as a yellow gum.
1H NMR (400MHz, methanol-d)4)δ ppm 1.04-1.22(m，3 H)2.43(s，3 H)3.96-4.24(m，2 H)4.35-4.55(m，1 H)5.03-5.17(m，1 H)6.13(s，2 H)7.12-7.47(m，6 H)，LCMS Rt＝3.29m/z 363[MH]+
Preparation 96
[ 2-amino-6- (2-fluoro-phenyl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
2-fluoro-phenylboronic acid (34mg), copper (I) thiophene-2-carboxylate (79mg), and bis (triphenylphosphine) palladium dichloride (19mg) were added to a microwave vial. The bottle was then purged with nitrogen and sealed. A solution of (2-amino-6-methylsulfanyl-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (42mg) in anhydrous THF (0.5ml) was then added to the bottle and the mixture was heated under microwave irradiation (CEM) at 100 ℃ for 10 minutes. The reaction mixture was then diluted with methanol and filtered directly over a cation exchange cartridge (Bakerbond, sulfonic acid bound phase, 1g) using Arbocel. The cartridge was washed with methanol (2 x 5ml) to remove impurities and then the product was liberated by elution with ammonia in methanol (2M, 5 ml). The desired eluted fractions were combined and evaporated to give the title compound (31mg) as a yellow gum.
1H NMR (400MHz, chloroform-d) δ ppm 1.03-1.31(m, 3H) 4.16(s, 2H) 4.91(br.s., 1H) 6.20(s, 2H) 6.95(s, 1H) 7.07(dd, J ═ 11.71, 8.20Hz, 1.20 Hz, 1H)7.16-7.22(m，1H)7.21-7.42(m，7H)7.84-7.93(m，1H)，LCMSRt＝3.50m/z 411[MH]+
Preparation 97
[ 2-amino-6- (3-fluoro-phenyl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 96 using 3-fluoro-phenylboronic acid to give the product as a yellow gum (18 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.08-1.35(m, 3H) 4.19(s, 2H) 5.29(br.s., 2H) 6.29(s, 2H) 6.66(s, 1H) 7.04-7.17(m, 1H) 7.28-7.40(m, 7H) 7.43-7.55(m, 1H), LCMS Rt＝3.53m/z 411[MH]+
Preparation 98
[ 2-amino-6- (4-fluoro-phenyl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 96 using 4-fluoro-phenylboronic acid to give the product as a yellow gum (28 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.19(s, 3H) 4.19(s, 2H) 5.33(br.s., 2H) 6.31(s, 2H) 6.63(s, 1H) 7.03-7.13(m, 2H) 7.29-7.38(m, 5H)7.68-7.77(m, 2H), LCMS Rt＝3.52m/z 411[MH]+
Preparation 99
(2-amino-6-methoxy-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
Sodium hydride (9mg) was carefully dissolved in methanol (0.5ml) and the solution was then added to a solution of (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (50mg) in THF (0.5ml) at room temperature. The reaction was stirred under nitrogen for 2 hours and then the solution was directly pre-adsorbed onto silica gel. The mixture was purified by Isco company chromatography eluting with ethyl acetate: heptane, increasing the gradient linearly from 20:80 to 60:40 over several column volumes. The desired eluted fractions were combined and evaporated to give the title compound (23mg) as a yellow gum.
1H NMR (400MHz, chloroform-d) δ ppm 1.15(s, 3H) 3.84(s, 3H)3.97-4.32(m, 2H) 4.42(d, J ═ 15.05Hz, 1H) 5.21(d, J ═ 15.05Hz, 1H) 5.77(s, 1H) 6.61(br.s., 2H) 7.23-7.38(m, 5H), LCMS Rt＝3.19m/z347[MH]+
Preparation 100
(2-amino-6-ethoxy-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 99, using ethanol to give the product as an off-white solid (31 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.10-1.28(m, 3H) 1.31(t, J ═ 7.03Hz, 3H)3.97-4.32(m, 4H) 4.43(d, J ═ 15.83Hz, 1H) 5.20(d, J ═ 15.83Hz, 1H)5.70-5.86(m, 1H) 6.61(br.s., 2H) 7.26-7.36(m, 5H), LCMS r.s., 2H) 7.10-7.36 (m, 5H), LCMS r.st＝3.36m/z 361[MH]+
Preparation 101
(2-amino-6-propoxy-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 99, using propanol to give the product as an off-white solid (30 mg).
1H NMR (400MHz, chloroform-d) δ ppm 0.95(t, J ═ 7.42Hz, 3H) 1.07-1.33(m, 3H)1.62-1.79(m, 2H) 3.95-4.29(m, 4H) 4.44(d, J ═ 15.63Hz, 1H) 5.20(d, J ═ 15.63Hz, 1H) 5.67-5.90(m, 1H) 6.61(br.s., 2H) 7.25-7.41(m, 5H), LCMS R ═ 7.42Hz, 3H) t＝3.51m/z 375[MH]+
Preparation 102
(2-amino-6-methylamino-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
Methylamine (40% aqueous solution) (0.055ml) was added to a solution of (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (50mg) in THF (0.5ml) at room temperature. The reaction was stirred under nitrogen for 16 hours and then the solution was directly pre-adsorbed onto silica gel. The mixture was purified by Isco company chromatography eluting with ethyl acetate: heptane, increasing the gradient linearly from 20:80 to 60:40 over several column volumes. The desired eluted fractions were combined and evaporated to give the title compound (30mg) as a yellow gum.
1H NMR (400MHz, chloroform-d) delta ppm 1.04-1.40(m, 3H) 2.82(s, 3H) 3.97-4.33(m, 3H) 4.75-5.04(m, 1H) 5.22-5.49(m, 2H) 6.79(br.s., 2H) 7.28-7.39(m, 5H), LCMS Rt＝2.83m/z 346[MH]+
Preparation 103
(2-amino-6-ethylamino-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 102 using ethylamine (70% in water) (0.051ml) to give the product as a yellow gum (35 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.05-1.37(m, 6H) 3.23(s, 2H) 3.93-4.35(m, 3H) 4.84(s, 1H) 5.17-5.52(m, 2H) 6.75(br.s., 2H) 7.29-7.42(m, 5H), LCMS R t＝3.0m/z 360[MH]+
Preparation 104
(2-amino-6-propylamino-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 102, using 1-propylamine (0.052ml) to give the product as a yellow gum (35 mg).
1H NMR (400MHz, chloroform-d) δ ppm 0.91(t, J ═ 7.42Hz, 3H) 1.06-1.38(m, 3H)1.44-1.55(m, 2H) 3.14(s, 2H) 3.93-4.30(m,3 H)4.75-5.07(m，1H)5.22-5.48(m，2 H)6.77(br.s.，2 H)7.28-7.37(m，5 H)，LCMS Rt＝3.16m/z 374[MH]+
preparation 105
(2-amino-6-butylamino-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 102 using n-butylamine (0.063ml) to give the product as a yellow gum (39 mg).
1H NMR (400MHz, chloroform-d) δ ppm 0.92(t, J ═ 7.42Hz, 3H) 1.12-1.37(m, 5H)1.41-1.51(m, 2H) 3.17(s, 2H) 3.98-4.31(m, 3H) 4.84(s, 1H) 5.26-5.47(m, 2H) 6.75(br.s., 2H) 7.28-7.36(m, 5H), LCMS Rt＝3.31m/z 388[MH]+
Preparation 106
[ 2-amino-6- (2-methoxy-ethylamino) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
The title compound was prepared as per the example in preparation 102 using 2-methoxyethylamine (1.0ml) and (2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (150mg) to give the product as a yellow gum (121 mg).
1H NMR(400MHz，DMSO-d6)δ ppm 1.21(t，3 H)，3.35(s，3 H)，3.45(m，4H)，4.20(q，2 H)，5.25(br，s，2 H)，7.20-7.35(m，5 H)，LRMS m/z(API)390[MH]+，388[MH]-。
Preparation 107
(2, 6-dibromo-3-nitro-pyridin-4-yl) - (6-trifluoromethyl-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 68 using potassium carbonate (1.50g), (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (2.0g), 5- (chloromethyl) -2-trifluoromethylpyridine (1.06g) and sodium iodide (0.98g) in acetone (40 ml). This gave a yellow oily product (2.79 g).
1H NMR(400MHz，DMSO-d6)δ ppm 1.20(t，3 H)，4.20(q，2 H)，4.85(s，2H)，7.28(s，1 H)，7.71(d，1 H)，7.88(dd，1 H)，8.61(d，1 H)
Preparation 108
(2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 69 using (2, 6-bromo-3-nitro-pyridin-4-yl) - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (2.75g), aqueous ammonia (11ml) and 2-methyl-THF (11 ml). This gave a yellow oily product (1.92 g).
1H NMR(400MHz，DMSO-d6)δ ppm 1.21(t，3 H)，4.11(q，2H)，4.95(br，s，2 H)，6.41(br，s，2 H)，6.60(s，1 H)，7.65(d，1H)，7.95(m，1 H)，8.65(m，1 H)
Preparation 109
(3-cyano-benzyl) - (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 68 using potassium carbonate (1.50g), (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (2.0g), 3-chloromethyl-benzonitrile (0.82g) and sodium iodide (0.98g) in acetone (40 ml). This gave a yellow oily product (2.62 g).
1H NMR(400MHz，DMSO-d6)δ ppm 1.20(t，3 H)，4.15(q，2H)，4.80(br，s，2 H)，7.15(s，1 H)，7.5(m，2 H)，7.61(m，1 H)，7.65(m，1H)
Preparation 110
(2-amino-6-bromo-3-nitro-pyridin-4-yl) - (3-cyano-benzyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 68 using (3-cyano-benzyl) - (2, 6-dibromo-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (2.32g), aqueous ammonia (10.2ml) and 2-methyl-THF (10.2 ml). This gave a yellow solid as product (1.51 g).
1H NMR(400MHz，DMSO-d6)δ ppm 1.20(t，3 H)，4.21(q，2H)，4.95(br，s，2 H)，6.35(br，s，2 H)，6.59(s，1 H)，7.5(m，1H)，7.6(m，3 H)
Preparation 111
(2-amino-3-nitro-6-oxazol-2-yl-pyridin-4-yl) - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using oxazole (30mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) and bis (triphenylphosphine) palladium dichloride (30mg) to give the product as a yellow solid (61 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.10-1.31(m, 3H) 4.02-4.30(m, 2H) 5.07(s, 2H) 6.39(s, 2H) 7.32(s, 1H) 7.35(s, 1H) 7.69(d, J ═ 7.82Hz, 1H) 7.84(s, 1H) 8.01(d, J ═ 5.47Hz, 1H) 8.70(s, 1H), LCMS R (R, g, c, m, g, c, H, c, g, H, c, gt＝3.02m/z 453[MH]+
Preparation 112
[ 2-amino-6- (4-methyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-methyl-oxazole (36mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (6-trifluoromethyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (100mg) and bis (triphenylphosphine) palladium dichloride (30mg) to give the product as a yellow solid (44 mg).
1H NMR (400MHz, chloroform-d) delta ppm 1.10-1.25(m, 3H) 2.28(s, 3H) 4.09-4.25(m, 2)H)5.06(s，2 H)6.41(s，2 H)7.25(s，1 H)7.55(d，J＝1.17Hz，1 H)7.69(d，J＝8.21Hz，1 H)8.01(d，J＝7.03Hz，1H)8.70(s，1 H)，LCMS Rt＝3.13m/z 467[MH]+
Preparation 113
(2-amino-3-nitro-6-oxazol-2-yl-pyridin-4-yl) - (3-cyano-benzyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using oxazole (33mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (3-cyano-benzyl) -carbamic acid ethyl ester (100mg) and bis (triphenylphosphine) palladium dichloride (33mg) to give the product as a yellow solid (67 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.20(t, J ═ 7.03Hz, 3H) 4.17(s, 2H) 4.99(s, 2H) 6.36(s, 2H) 7.26(s, 1H) 7.34(s, 1H) 7.46(t, J ═ 7.62Hz, 1H) 7.59(d, J ═ 7.42Hz, 1H) 7.62-7.68(m, 2H) 7.82(s, 1H), LCMSR (LCMSR), LCMSR (r), c, g, c, gt＝2.93m/z 409[MH]+
Preparation 114
[ 2-amino-6- (4-methyl-oxazol-2-yl) -3-nitro-pyridin-4-yl ] - (3-cyano-benzyl) -carbamic acid ethyl ester
The title compound was prepared as for example in preparation 70 using 4-methyl-oxazole (40mg), (2-amino-6-bromo-3-nitro-pyridin-4-yl) - (3-cyano-benzyl) -carbamic acid ethyl ester (100mg) and bis (triphenylphosphine) palladium dichloride (30mg) to give the product as a yellow solid (65 mg).
1H NMR (400MHz, chloroform-d) δ ppm 1.10-1.25(m, 3H) 2.28(s, 3H) 4.10-4.23(m, 2H) 4.99(s, 2H) 6.39(s, 2H) 7.20(s, 1H) 7.46(t, J ═ 7.62Hz, 1H) 7.54(d, J ═ 1.17Hz, 1H) 7.60(d, J ═ 7.42Hz, 1H)7.63-7.69(m, 2H), LCMS R δ ppm t＝3.04m/z 423[MH]+
Preparation 115
4-amino-1-benzyl-6-bromo-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
(2-amino-6-bromo-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (50mg, 0.13mmol) was dissolved in AcOH (3 ml). Iron powder (43mg, 0.76mmol) was added and the mixture was stirred vigorously at room temperature for 24 hours. The reaction mixture was diluted with EtOAc (20ml) and water (10 ml). The mixture was filtered through celite and washed with EtOAc (20 ml). The layers were separated and the organic layer was washed with water (10ml), saturated NaHCO3The aqueous solution (2X 10ml) and brine (10ml) were washed with MgSO4Dried and concentrated in vacuo. The crude material was triturated in pentane, filtered and dried in vacuo at 40 ℃ to give the title compound as a beige solid (15 mg).
1H NMR (d 6-DMSO). delta.10.47 (br s, 1H), 7.36-7.26(m, 5H), 6.70(s, 1H), 6.05(br s, 2H), 4.94(s, 2H); LRMS (APCI and ES) m/z 319/321[ MH]+。
Preparation 116
Benzyl- (2, 3-diamino-6-pyrazol-1-yl-pyridin-4-yl) -carbamic acid ethyl ester
(2-amino-3-nitro-6-pyrazol-1-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (65.4mg, 0.171mmol) was dissolved in methanol (5ml) and H80 psi at room temperature2Hydrogenation was then carried out over Raney nickel (25mg) for 1 hour. The reaction mixture was filtered through an Arbocel short plug and the filtrate was evaporated in vacuo to yield the title compound as a 58mg brown residue.
1H NMR(CDCl3)δ 1.15(m，3H)，2.90(s br，2H)，3.50(m，2H)，4.12(d，2H)，4.20(s br，2H)，6.28(s br，1H)，7.10(s br，1H)，7.20-7.35(m，5H)，7.85(s，1H)，8.30(s，1H)；LRMS(ES+)m/z 353(MH+)。
Preparation 117
(2-amino-3-nitro-6- [1, 2, 4] triazol-1-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
The title compound was prepared according to the examples in preparation 116.
1H NMR(CDCl3) δ 1.20(t, 3H), 4.19 (quartet, 2H), 5.00(sbr, 2H), 6.37(sbr, 2H), 7.09(s, 1H), 7.27-7.37(m, 5H), 8.03(s, 1H), 9.00(s, 1H); LRMS (ES)+)m/z 384(MH+)。
Preparation 118
(ai) (2-amino-3-nitro-6- [1, 2, 3] triazol-2-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester and (bi) (2-amino-3-nitro-6- [1, 2, 3] triazol-1-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
(2-amino-6-chloro-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (100mg, 0.285mmol) was dissolved in acetonitrile (10ml) and 1, 2, 3-triazole (39.4mg, 0.570mmol), potassium carbonate (78.8mg, 0.570mmol) were added. The yellow solution was stirred at 70 ℃ under nitrogen for 18 hours. The resulting dark orange reaction mixture was evaporated in vacuo and the residue was dissolved in EtOAc and extracted with water. The organic extracts were combined, dried over anhydrous magnesium sulfate and concentrated in vacuo. The 2 structural isomers were separated by auto-purification (chiralpak column, 50:50 methanol: ethanol) to yield 48.1mg of (ai) (2-amino-3-nitro-6- [1, 2, 3] triazol-2-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester and 34.9mg of (bi) (2-amino-3-nitro-6- [1, 2, 3] triazol-1-yl-pyridin-4-yl) -benzyl-carbamic acid ethyl ester all as yellow solids.
1H NMR(CDCl3)(ai)δ 1.19(t，3H)，4.18(d，2H)，5.00(s br，2H)，6.62(sbr，2H)，7.25-7.36(m，6H)，7.87(s，1H).LRMS(ES+)m/z 384(MH+)。
1H NMR(CDCl3)(bi)δ 1.21(t，3H)，4.20(q，2H)，5.02(s br，2H)，6.35(sbr，2H)，7.24-7.40(m，6H)，7.77(s，1H)，8.40(d，1H).LRMS(ES+)m/z384(MH+)。
Preparation 119
[ 2-amino-6- (4-fluoro-pyrazol-1-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
(2-amino-6-chloro-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (100mg, 0.285mmol), 4-fluoropyrazole (24.5mg, 0.285mmol) and potassium carbonate (118mg, 0.855mmol) were stirred together in 5ml acetonitrile under nitrogen. The reaction was heated to reflux temperature for 3 hours and then cooled to ambient temperature overnight.
The solvent was evaporated and the dichloromethane soluble fraction of the residue was purified by silica chromatography eluting with 1% methanol in dichloromethane. The fractions containing pure material were combined with Rf of 0.66 in the same eluent and evaporated to give the title compound as a yellow gum (106 mg).
1H NMR(CDCl3400MHz) δ 1.20 (broad singlet, 3H), 4.17 (broad singlet, 2H), 4.95 (broad doublet, 2H)6.38 (broad singlet, 2H)7.15(s, 1H)7.35(m, 5H)7.58(d, 1H)8.23(d, 1H); LRMS (ES)+)m/z 401(MH+)
The following compounds were prepared using the same method as described in preparation 119.
Preparation 120
[ 2-amino-6- (3, 5-dimethyl-pyrazol-1-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
1H NMR(CDCl3400MHz) δ 1.19 (broad singlet, 3H), 2.24(s, 3H)2.62(s, 3H)4.16 (broad singlet, 2H), 4.94 (broad doublet, 2H)5.98(s, 1H)6.36 (broad singlet, 2H)7.21(s, 1H)7.32(m, 5H); LRMS (ES) +)m/z 411(MH+)。
Preparation 121
[ 2-amino-6- (4-methyl-pyrazol-1-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
1H NMR(CDCl3400MHz) δ 1.20 (broad singlet, 3H), 2.14(s, 3H)4.16 (broad singlet, 2H), 4.95 (broad doublet, 2H)6.39 (broad singlet, 2H)7.16(s, 1H)7.36(m, 5H).7.53(s, 1H)8.15(s, 1H); LRMS (ES)+)m/z 397(MH+)
Preparation 122
[ 2-amino-6- (3-trifluoromethyl-pyrazol-1-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
1H NMR(CDCl3400MHz) δ 1.22 (broad singlet, 3H), 4.20 (broad singlet, 2H), 5.00 (broad doublet, 2H), 6.33 (broad singlet, 2H)6.69(d, 1H), 7.22(s, 1H), 7.35(m, 5H), 8.45(d, 1H); LRMS (ES)+)m/z451(MH+)。
Preparation 123
[ 2-amino-6- (5-methyl-3-trifluoromethyl-pyrazol-1-yl) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
1H NMR(CDCl3400MHz) δ 1.22 (broad singlet, 3H), 2.69(s, 3H), 4.20 (broad singlet, 2H), 5.00 (broad singlet, 2H), 6.29 (broad singlet, 2H), 6.41(s, 1H), 7.19(s, 1H), 7.33(m, 5H); LRMS (ES)+)m/z 465(MH+)。
Preparation 124
{ 2-amino-6- [4- (2-hydroxy-ethyl) -pyrazol-1-yl ] -3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
1H NMR(CDCl3400MHz) δ 1.20 (broad peak, 3H), 2.78(t, 2H), 3.85(q, 2H)4.18 (broad singlet, 2H), 4.98 (broad singlet, 2H), 6.37 (broad singlet, 2H), 7.17(s, 1H), 7.33(m, 5H), 7.61(s, 1H), 8.28(s, 1H); LRMS (ES) +)m/w 427(MH+)
Preparation 125
1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylic acid- (1-hydroxyimino-ethyl) -amide
In a reaction flask, CDI (52mg, 0.32mmol) was added to 1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c)]Pyridine-6-carboxylic acid (80mg, 0.21mmol) was dissolved in Hunnigs base (83. mu.l, 0.64mmol) in anhydrous DMF. The bottle was sealed and the mixture was stirred at room temperature for 15 minutes. N-Hydroxyacetamidine (24mg, 0.32mmol) was added and the mixture was heated to 60 ℃ in an aluminum block. The mixture was stirred at this temperature for 3 hours, then cooled and concentrated in vacuo. The residue was partitioned between EtOAc (15ml) and water (5 ml). The layers were separated and the organic phase was washed with brine (5 ml). At this point, some solids precipitated and were placed in a separatory funnel. It was washed into the organic phase with MeOH, and the organics were dried (over MgSO4) And evaporated to give the final product as a white solid (120mg, 130%). This material may contain inorganic materials, but is useful for the next step in terms of purification.
1H NMR CD3OD δ 1.94(s，3H)4.54(s，2H)5.16(s，2H)7.16-7.46(m，11H)。
LCMS Rt＝3.47m/z 431[MH]+
Preparation 126
N2, N2-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine
2, 6-dibromo-4-amino-5-nitro-pyridine (3.52g) was dissolved in 2-methyl THF (40ml) and the solution was cooled to <5 ℃ in an ice bath. A solution of dibenzylamine (2.39ml) and triethylamine (2.48ml) in 2-methyl THF (20ml) was added dropwise to the dibromopyridine solution, and the reaction mixture was warmed to room temperature and stirred under a nitrogen atmosphere for 16 hours. Dibenzylamine (684. mu.l) and triethylamine (496. mu.l) were further added, and the mixture was stirred at room temperature for a further 5 hours, then dibenzylamine (684. mu.l) and triethylamine (496. mu.l) were further added, and the mixture was stirred at room temperature for a further 16 hours.
Transferring the mixture toInto a separatory funnel, and then water (60ml) was added. The layers were separated and the aqueous solution re-extracted with EtOAc (60 mL). The combined organics were dried (over MgSO4) And evaporated to an orange gum. The gum was crystallized from MeOH: water (90:10,. about.200 ml) to give a solid, which was filtered and washed with MeOH: water (90:10) then dried under vacuum to give the title compound as an orange crystalline solid (3.6 g).
1H NMR(CDCl3，400MHz)δ 4.45(s，4H)，5.95(br，s，2H)，6.22(s，1H)，7.05-7.15(m，4H)，7.21-7.35(m，6H)。LCMS Rt＝3.73m/z 415[MH]+
Preparation 127
N2, N2-dibenzyl-6-bromo-N4- (6-methyl-pyridin-3-ylmethyl) -3-nitropyridine-2, 4-diamine
Potassium tert-butoxide (448mg) was added dropwise under a nitrogen atmosphere to a cold solution (-18 ℃ -salt/ice bath) of N2, N2-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (1500mg, 3.63mmol) in THF (40 ml). After addition of the base, the solution turned from yellow to dark red/orange. The solution was stirred in the cooling bath for 5 minutes.
5-bromomethyl-2-methyl-pyridine hydrobromide (1160mg) in saturated NaHCO3The solution (20ml) was partitioned between 2-Me THF (20 ml). The phases were separated and the aqueous solution re-extracted with 2-MeTHF (20 ml). The combined organics were dried (over MgSO4) Then added dropwise to the aminopyridine and KOtBu mixture with a dropping funnel. The color changed from red/orange to yellow/orange. The mixture was slowly warmed to room temperature in a cooling bath and then stirred at room temperature under nitrogen atmosphere for 48 hours. The reaction mixture was cooled again to-18 deg.C (ice/salt bath) and then KOtBu (102mg), tetra-n-butylammonium iodide (670mg) were added in that order. The cooling bath was removed and the mixture was warmed to room temperature and then stirred for an additional 4 hours. The reaction mixture was pre-adsorbed directly onto silica, then column chromatographed through an Isco company silica column (80g, Redisep) eluting with EtOAc: heptane, linearly increasing from 40:60 to 80: 20. The desired eluting components were combined and evaporated to give the title compound as a yellow solid (0.99 g).
1H NMR(CDCl3，400MHz)δ 2.58(s，3H)，4.38(d，2H)，4.45(s，4H)，6.19(s，1H)7.05-7.15(m，4H)，7.18(d，1H)，7.21-7.35(m，6H)，7.49(dd，1H)，7.99(t，1H)，8.42(d，1H)。LCMS Rt＝3.12m/z 520[MH]+
Preparation of 128
N2, N2-dibenzyl-N4- (6-methyl-pyridin-3-ylmethyl) -3-nitro-6-oxazol-2-yl-pyridine-2, 4-diamine
Butyllithium (2.9ml) was added dropwise to a stirred solution of oxazole (0.251ml) in THF (5ml) at-78 ℃ (dry ice/acetone bath), maintaining the rate of addition such that the reaction temperature did not exceed-60 ℃. The solution was stirred at this temperature for 10 minutes, then a solution of zinc chloride (1.56g) in THF (7ml) was added dropwise. The solution was stirred at-78 ℃ for 15 minutes, then the cooling bath was removed and the reaction mixture was warmed to room temperature. An aliquot (1.2ml) of the zinc oxazole solution was added by syringe to a pre-sealed nitrogen purged microwave vial (Biotage, 0.5-2.0ml) containing N2, N2-dibenzyl-6-bromo-N4- (6-methyl-pyridin-3-ylmethyl) -3-nitropyridine-2, 4-diamine (90mg) and bis (triphenylphosphine) palladium dichloride (24 mg). The vial was heated at 130 ℃ for 15 minutes under microwave irradiation (Biotage Initiator 8). Nine additional aliquots of the zinc oxazole solution were reacted with bromopyridine in the same manner in a microwave.
All reaction mixtures were combined and concentrated in vacuo to a brown gum. The gel was partitioned between EtOAc (20ml) and 2M ammonia solution (20 ml). The aqueous solution was re-extracted with EtOAc (20ml) and the combined organics were washed with brine (20ml) and then dried (MgSO 2) 4) And evaporated to brown gum. The crude product was purified on an Isco company silica cartridge (120g, Redisep) eluting with EtOAc in a linear increase from 80:20 to 100:0 over a 6 column volume gradient, then 18 column volumes at 100% EtOAc isocratic. The desired eluting components are combined and evaporatedTo give the title compound (0.43g) as a yellow foamy solid.
1H NMR(CDCl3，400MHz)δ 2.60(s，3H)，4.55(d，2H)，4.60(s，4H)，6.99(s，1H)7.05-7.15(m，4H)，7.20-7.22(d，2H)，7.22-7.35(m，6H)，7.59(dd，1H)，7.79(s，1H)，8.09(t，1H)，8.55(d，1H)。LCMS Rt＝2.87m/z 507[MH]+
Preparation 129
N2, N2-dibenzyl-N4- (6-methyl-pyridin-3-ylmethyl) -6-oxazol-2-yl-pyridine-2, 3, 4-triamine
N2, N2-dibenzyl-N4- (6-methyl-pyridin-3-ylmethyl) -3-nitro-6-oxazol-2-yl-pyridine-2, 4-diamine (425mg) was dissolved in THF (60ml) followed by addition of MeOH (60 ml). The solution was hydrogenated over Raney nickel (40mg) under a hydrogen atmosphere (80psi) for 1 hour. The reaction mixture was filtered through a pad of celite and then evaporated to give the title compound as a yellow crude gum which was used directly in the next step.
LCMS Rt＝2.42m/z 477[MH]+。
Preparation 130
(2, 6-dichloro-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester
A solution of ethyl chloroformate (2.75ml) in 2-methyl THF (10ml) was added dropwise to a cold (ice bath) solution of 2, 6-dichloro-4-amino-5-nitro-pyridine (5.00g) and triethylamine (4.02ml) in 2-methyl THF (50 ml). The rate of addition should be such that the reaction temperature does not rise to 5 ℃. After addition of ethyl chloroformate, a precipitate formed. The suspension was warmed to room temperature and then stirred under nitrogen for 16 hours. The suspension was transferred to a separatory funnel and water (50ml) was added. The layers were separated and the organics were washed with brine (50ml) and then dried (over MgSO 4) And evaporated to an orange gum which solidified to a yellow solid upon standing. The solid was recrystallized from MeOH: water (70:30) to give the title compound as white needles, which was collected by filtration (6.7 g).
1H NMR(CDCl3，400MHz)δ 1.25(t，3H)，4.31(q，2H)，8.10(br，s，1H)，8.40(s，1H)。LCMS Rt＝4.16m/z 280[MH]+
Preparation 131
Benzyl- (2, 6-dichloro-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester
Benzyl bromide (2.33ml) was added dropwise to a stirred suspension of (2, 6-dichloro-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (4.57g) in acetonitrile (40 ml). The mixture was stirred at room temperature under nitrogen for 16 hours. The mixture was concentrated in vacuo and then partitioned between EtOAc (50ml) and water (50 ml). The layers were separated and the organics were washed with saturated NH4Cl (50ml), water (50ml) and brine (50 ml). The organics were dried (over MgSO4) And evaporated to a yellow oil. This material was pre-adsorbed on silica, then column chromatographed on an Isco company silica column (330g, Redisep) eluting with EtOAc in a 1 Column Volume (CV) at 10:90 isocratic, then the gradient was linearly increased from 10:90 to 30:70 over 6 CVs. The desired eluting components were combined and evaporated to give the title compound as a yellow oil (6.05 g).
1H NMR(CDCl3，400MHz)δ 1.25(t，3H)，4.19(q，2H)，4.81(s，2H)，6.88(s，1H)，7.20-7.28(m，2H)，7.30-7.40(m，3H)。LCMS Rt＝3.62m/z372[MH]+。
Preparation 132
(2-amino-6-chloro-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester
Benzyl- (2, 6-dichloro-3-nitro-pyridin-4-yl) -carbamic acid ethyl ester (500mg) was dissolved in THF (3ml) in a reaction flask. Ammonia (7M in MeOH, 1ml) was added, the vial was sealed, and the reaction was stirred at room temperature for 48 hours. The reaction mixture was then pre-adsorbed directly onto silica and column chromatographed on an Isco company silica column (40g, Redisep) eluting with EtOAc: heptane, with a gradient increasing linearly from 10:90 to 40:60 over 10 column volumes. The desired eluting components were combined and evaporated to give the title compound as a yellow gum which solidified on standing (305 mg).
1H NMR(CDCl3，400MHz)δ 1.05(t，3H)，4.02(q，2H)，4.87(br，s，2H)，6.58(s，1H)，7.20-7.36(m，5H)，7.61(br，s，1H)。LCMS Rt＝3.24m/z351[MH]+
Preparation 133
[ 2-amino-6- (2-methoxy-ethoxy) -3-nitro-pyridin-4-yl ] -benzyl-carbamic acid ethyl ester
Sodium hydride (21mg) was added dropwise to 2-methoxyethanol (0.5 ml). The resulting solution was added dropwise to a solution of (2-amino-6-chloro-3-nitro-pyridin-4-yl) -benzyl-carbamic acid ethyl ester (100mg) in THF (1.0 ml). The reaction mixture turned from a yellow solution to a dark red/orange solution and was stirred at room temperature for 1 hour. The orange mixture was concentrated in vacuo and then washed with EtOAc (10mL) and saturated NH4The Cl solution (10ml) was partitioned between. The layers were separated and the organics were washed with water (10ml) and brine (10ml) and then dried (over MgSO 4) And evaporated to give the title compound (111mg) as a crude yellow gum. This was used directly in the next step without further purification.
LCMS Rt＝3.13m/z 391[MH]+
Preparation of 134
4-benzylamino-3-nitro-pyridin-2-ol
4-chloro-3-nitro-2-pyrrolidone (5g, 28.65mmol) was suspended in 150ml of acetonitrile, and then benzylamine (3.15ml, 28.65mmol), potassium carbonate (4g, 28.65mmol) were added in this order, and the mixture was stirred at 60 ℃ overnight. The solvent was removed in vacuo and the residue was suspended in water (200 ml). Add 2M HCl solution until pH-6. The precipitate was filtered and dried in vacuo to yield 4.75g of the title compound as a beige solid. Harvest from mother liquor 2 nd time yielded 1.4g of the title compound.
1H NMR (d6 DMSO) δ: 11.20 (broad singlet, 1H), 9.35(t, 1H), 7.40-7.20(m, 6H), 5.85(d, 1H), 4.60(d, 2H); LRMS (ES)+)m/z 246[MH]+
Preparation of 135
Benzyl- (2-chloro-3-nitro-pyridin-4-yl) -amine
4-benzylamino-3-nitro-pyridin-2-ol (6.15g, 25.07mmol) was suspended in 100ml acetonitrile, then phosphorus oxychloride (12ml, 125.40mmol), tetraethylammonium chloride (4.15g, 25.07mmol) were added sequentially, and the mixture was stirred at 85 ℃ overnight. The solvent was removed in vacuo and the residue was suspended in water (300ml) and extracted with dichloromethane (2 × 200 ml). The organic layer was dried over magnesium sulfate and the solvent was removed in vacuo to yield 5.9g of the title compound as a yellow solid.
1H NMR(CDCl3) δ: 8.0(d, 1H), 7.40-7.20(m, 5H), 6.9 (broad singlet, 1H), 6.60(d, 1H), 4.5(d, 2H); LRMS (ES)+)m/z 264[MH]+
Preparation 136
N-2, N-2, N-4, tribenzyl-3-nitro-pyridine-2, 4-diamine
Benzyl- (2-chloro-3-nitro-pyridin-4-yl) -amine (3.95g, 14.99mmol) was suspended in 50ml acetonitrile, then dibenzylamine (2.9ml, 14.99mmol), potassium carbonate (2g, 14.99mmol) were added sequentially and the mixture was stirred at 80 ℃ overnight. The solvent was removed in vacuo and the residue was suspended in water (100ml) and extracted with ethyl acetate (2 × 100 ml). The organic layer was dried over magnesium sulfate and the solvent was removed in vacuo. The crude residue was purified by silica gel column chromatography using 15% ethyl acetate in pentane to give 6g of the title compound as a yellow oil.
1H NMR (d6 DMSO) δ: 8.1(s broad, 1H), 7.9(d, 1H), 7.40-7.10(m, 15H), 6.10(d, 1H), 4.55(s, 4H), 4.5(d, 2H); LRMS (ES)+)m/z 425[MH]+
Preparation 137
N-2, N-2, N-4-tribenzyl-pyridine-2, 3, 4-triamine
N-2, N-2, N-4, tribenzyl-3-nitro-pyridine-2, 4-diamine (6g, 14.13mmol) was suspended in 150ml of ethanol, Raney nickel (1.2g, 20 wt%) was added, and the mixture was stirred at room temperature under 50psi of hydrogen for 3 hours. Upon completion, the mixture was filtered with Arbocel (lignocellulose) and the solvent was removed in vacuo to yield 5g of the title compound as a pale purple gum.
1H NMR(CDCl3) δ: 7.80(d, 1H), 7.40-7.15(m, 15H), 6.35(d, 1H), 4.35(d, 2H), 4.15(s, 4H), 3.5-3.25 (broad singlet, 2H); LRMS (ES)+)m/z 395[MH]+
Preparation 138
1-benzyl-4-dibenzylamino-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one
N-2, N-2, N-4, tribenzyl-pyridine-2, 3, 4-triamine (5g, 12.67mmol) was dissolved in 100ml acetonitrile, then 1, 1' -carbonyldiimidazole (3g, 19.701mol) was added and the reaction was stirred at 80 ℃ overnight. The mixture was cooled to room temperature and the precipitate was filtered and washed with acetonitrile, then dried in vacuo to yield 4.3g of the title compound as a pale purple solid.
1H NMR(d6 DMSO)δ：10.9(s，1H)，7.75(d，1H)，7.40-7.10(m，15H)，6.70(d，1H)，4.9(s，2H)，4.6(s，4H)；LRMS(ES+)m/z 421[MH]+。
Preparation 139
1-benzyl-7-bromo-4-dibenzylamino-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one
1-benzyl-4-dibenzylamino-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one (1g, 2.4mmol) was suspended in 20ml of acetic acid, followed by dropwise addition of sodium acetate (195mg, 2.4mmol), bromine (456mg, 2.85 mmol). The mixture was stirred at room temperature for 15 minutes. A thick precipitate formed. The mixture was diluted in water (50ml) and the solid was filtered and washed with water. It was then diluted in ethyl acetate (20ml), dried over magnesium sulfate and the solvent removed in vacuo to give the title compound as a pale orange solid, 1.29 g.
1H NMR(d6 DMSO)δ：11.5(s，1H)，7.85(s，1H)，7.40-7.10(m，15H)，5.30(s，2H)，4.55(s，4H)；LRMS(ES+)m/z 499，501[MH]+
Preparation 140
1-benzyl-4-dibenzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5, c ] pyridine-7-carboxylic acid methyl ester
1-benzyl-7-bromo-4-dibenzylamino-1, 3-dihydro-imidazo [4, 5, c ] pyridin-2-one (500mg, 1mmol) was suspended in 30ml of methanol, then triethylamine (203mg, 2mmol), (1, 1' -bis (diphenylphosphino) ferrocene) dichloropalladium (82mg, 0.1mmol) was added in that order, and the mixture was stirred at 100 ℃ under 100psi CO overnight. The mixture was cooled to room temperature, filtered through arbor 1 and washed with methanol. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography using 1% methanol in dichloromethane to yield 21mg of the title compound as a white solid.
1H NMR(CDCl3) δ: 8.4(s, 1H), 8.0 (broad singlet, 1H), 7.40-7.00(m, 15H), 5.45(s, 2H), 4.80(s, 4H), 3.7(s, 3H); LRMS (ES)+)m/z 479[MH]+
Preparation 141
1-benzyl-4-dibenzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5, c ] pyridine-7-carboxylic acid cyclopropylmethylamide
1-benzyl-4-dibenzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5, c)]Pyridine-7-carboxylic acid methyl ester (50mg, 0.1mmol) was suspended in 2ml of (aminomethyl) cyclopropane, andthe mixture was stirred at 120 ℃ overnight. The excess amine was removed in vacuo and the gum was dissolved in water (20ml) and ethyl acetate (50ml), the organic layer was separated, MgSO 4Dried and the solvent removed in vacuo. The residue was purified by silica gel column chromatography using 5% methanol in dichloromethane to give 10mg of the title compound as a yellow gum; LRMS (ES)+)m/z 518[MH]+
Preparation 142
6-chloro-2, 4-dihydroxy-5-methyl-pyridine
Malonyl chloride (10g, 71mmol) was mixed with propionitrile (12ml) and stirred at room temperature under nitrogen for 16 h. 50mL of dioxane was added to the resulting heterogeneous mixture and the resulting precipitate was collected by filtration and washed with cold dioxane. The collected solid was dissolved in a few ml of methanol and reprecipitated with dioxane. The solid was collected by filtration, washed with dioxane and dried in vacuo to give the title compound as a white solid (6g, 53%); LRMS: (ES)+)m/z 160[MH]+。
Preparation 143
6-chloro-2, 4-dihydroxy-5-methyl-3-nitro-pyridine
6-chloro-2, 4-dihydroxy-5-methyl-pyridine (500mg, 3.1mmol) was dissolved in acetic acid (1ml), cooled to 0 ℃ and nitric acid (4ml) was added dropwise with stirring. After the addition was complete, the ice bath was removed and the reaction mixture was warmed to room temperature over 16 hours. Ice was added to the mixture to give a precipitate, which was collected by filtration and dried in vacuo to give the title compound as a yellow solid (180mg, 28%); LRMS: (ES) +)m/z 205[MH]+
Preparation 144
Malonic acid diphenyl ester
Malonic acid (11g, 106mmol) was mixed with phenol (20g, 212mmol) at 0 ℃ under nitrogen atmosphere and phosphorus oxychloride (11.5ml, 123mmol) was added dropwise to the solid mixture. The resulting mixture was stirred at 0 ℃ 5After a period of minutes, and then heating at reflux temperature for 5 hours, the solid melted and an orange solution formed. The reaction was cooled to room temperature, then poured onto 100ml of water and extracted with ether (3X 75 ml). The combined organics were washed with brine, over MgSO4The title compound was dried and concentrated in vacuo to an orange oil (27g, 99%).
1HNMR(CDCl3，400MHz)3.86(s，2H)，7.17(m，4H)，7.27(m，2H)，7.41(m，4H)；LRMS(ES)m/z 257[MH]+
Preparation 145
Cyclopentanone-tert-butylimine
Cyclopentanone (13.3ml, 150mmol) and tert-butylamine (47.4ml, 450mmol) were mixed in 110ml of diethyl ether under nitrogen and then cooled to-55 ℃ in a dry ice/acetonitrile bath. Titanium tetrachloride (8.2ml, 75mmol) was dissolved in 70ml pentane and added dropwise to the above solution, taking care to maintain the temperature at-40 ℃. The reaction was then stirred at-40 ℃ for 6 hours and then allowed to warm to room temperature overnight. The reaction mixture was filtered through a short plug of celite and washed with ether. The filtrate was evaporated in vacuo to yield 15.9g (76%) of the title product as a clear oil.
1HNMR(CDCl3，400MHz)1.26(s，9H)，1.67(m，2H)，1.82(m，2H)，2.29(t，2H)，2.36(t，2H)。
Preparation 146
6, 7-dihydro-5H 1-pyridine-2, 4-diol
Cyclopentanone-tert-butylimine (2.78g, 20mmol) and diphenyl malonate (5.12g, 20mmol) were mixed in 40ml of triglyme and heated at 100 ℃ for 4 hours and then at 200 ℃ for 2 hours. The reaction was then cooled to room temperature, poured into 200ml of diethyl ether and stored in a sealed flask in a refrigerator for 4 days. The resulting precipitate was filtered, washed with ether and dried in vacuo to give the title compound as a pale brown solid (1.45g, 50%).
1HNMR(CD3OD，400MHz)2.12(m，2H)，2.70(t，2H)，2.82(t，2H)，5.64(s，1H)。
Preparation 147
5, 6-dimethyl-pyridine-2, 4-diol
5, 6-dimethyl-4-hydroxy-2-oxo-2H-pyran (J.chem.Soc.PerkinTrans 1, 1980, 2272) (10g, 71mmol) was dissolved in 66ml dioxane and 33ml 0.88NH3To the solution and the mixture was refluxed for 3 hours. The resulting suspension was then cooled to room temperature overnight, the solid collected by filtration and dried in vacuo to yield the title compound as a white crystalline solid (6.5 g). The filtrate was concentrated to about 10ml in vacuo and the 2 nd harvest solid (1.0g) was collected by filtration. The two harvests were combined for the next synthesis step.
1H NMR(DMSO，400MHz)：δ 1.77(s，3H)，2.06(s，3H)，5.42(s，1H)；LRMSm/z(APCI+)140[MH]+。
Preparation 148
5, 6-dimethyl-3-nitro-pyridine-2, 4-diol
5, 6-dimethyl-pyridine-2, 4-diol (6.5g, 47mmol) was stirred in 30ml sulfuric acid and then cooled to 0 ℃ in an ice bath. Fuming nitric acid (10ml) was added dropwise and the mixture was stirred for 1 hour after the end of the addition. The reaction mixture was poured onto crushed ice and the resulting yellow solid was collected by filtration to obtain the title compound (3.9g, 46%).
1H NMR(MeOD，400MHz)：δ 2.04(s，3H)，2.31(s，3H)；LRMS m/z(APCI+)185[MH]+。
Preparation 149
2, 4-dichloro-5, 6-dimethyl-3-nitro-pyridine
5, 6-dimethyl-3-nitro-pyridine-2, 4-diol (3.9g, 21mmol) was dissolved in acetonitrile (150ml) and tetraethylammonium chloride was added(7.1g, 42mmol) and phosphorus oxychloride (19.9ml, 210mmol), and the whole was heated at 70 ℃ for 16 hours. The reaction mixture was poured into crushed ice and extracted with DCM (2 × 30 ml). The combined extracts were extracted with MgSO4Dried, filtered and concentrated in vacuo to give a brown solid. This solid was dissolved in 2ml DCM and washed with 2: 1 pentane: EtOAc eluted, filtered through a short plug of silica gel. The filtrate was then evaporated to give the title compound as a pale brown solid (3.5g, 75%).
1H NMR(CDCl3，400MHz)：δ 2.61(s，3H)，2.41(s，3H)
Preparation 150
Benzyl- (2-chloro-5, 6-dimethyl-3-nitro-pyridin-4-yl) -amine
2, 4-dichloro-5, 6-dimethyl-3-nitro-pyridine (2g, 9mmol) was dissolved in acetonitrile (100ml) and benzylamine (1.0ml, 9.5 mmol). Potassium carbonate (1.3g, 9.5mmol) was added in one portion and the whole was heated at 55 ℃ for 16 hours. The reaction mixture was diluted with EtOAc and washed with 50ml water. The aqueous solution was re-extracted with EtOAc, and the organics were combined and then MgSO4Dried and evaporated to a dark red residue. Use was made of 8: 1 pentane: the residue was purified by silica gel column chromatography with EtOAc as eluent to give the title compound as a bright orange solid (1.2g, 45%).
1H NMR(CDCl3，400MHz)：δ 2.09(s，3H)，2.47(s，3H)，4.25-4.27(d，2H)，4.52(bs，1H)，7.28-7.30(m，2H)，7.35-7.41(m，3H)LRMS m/z(APCI+)292[MH]+。
Preparation 151
N2, N2-diallyl-N4-benzyl-5, 6-dimethyl-3-nitro-pyridine-2, 4-diamine
Benzyl- (2-chloro-5, 6-dimethyl-3-nitro-pyridin-4-yl) -amine (1.2g, 4.1mmol) was dissolved in ethoxyethanol (60ml) and diisopropylethylamine (1.1ml, 6.2mmol) and diallylamine (0.76ml, 6.2mmol) were added in one portion. The reaction mixture was heated in a sealed vessel at 100 ℃ overnight and then concentrated in vacuo to an orange residue. This residue was directly purified by silica gel column chromatography using a gradient of 8:1 → 1:1 pentane: EtOAc as eluent to give the title compound as a bright orange oil (938mg, 65%).
1H NMR(CDCl3，400MHz)：δ 2.16(s，3H)，2.34(s，3H)，3.90-3.92(d，4H)，4.33-4.34(d，2H)，5.13-5.21(m，4H)，5.77-5.87(m，2H)，6.37-6.40(bt，1H)，7.35-7.28(m，5H)；LRMS m/z(APCI+)353[MH]+
Preparation 152
N2, N2-diallyl-N4-benzyl-5, 6-dimethyl-pyridine-2, 3, 4-triamine
N2, N2-diallyl-N4-benzyl-5, 6-dimethyl-3-nitro-pyridine-2, 4-diamine (828mg, 2.4mmol) was dissolved in ethanol (15ml) and 2N HCl (15ml) and iron powder (527mg, 9.6mmol) was added in one portion. The reaction mixture was heated at 70 ℃ for 2 hours, then cooled to room temperature and poured into 50ml of water. The resulting solution was neutralized with 1N NaOH solution to give a dark green suspension, which was extracted with EtOAc (2X 25ml), and the combined organics were extracted with MgSO 4Dried, filtered and evaporated to give the title compound as a dark green oil (559mg, 74%).
1H NMR(CDCl3，400MHz)：δ 1.95(s，3H)，2.32(s，3H)，3.73-3.74(d，4H)，4.21(s，2H)，5.07-5.23(m，4H)，5，86-5.96(m，2H)，7.26-7.32(m，5H)；LRMS m/z(APCI+)323[MH]+
Preparation 153
1-benzyl-4-diallylamino-6, 7-dimethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one
N2, N2-diallyl-N4-benzyl-5, 6-dimethyl-pyridine-2, 3, 4-triamine (559mg, 1.7mmol) was dissolved in acetonitrile (50ml) and 1, 1-carbonyldiimidazole (2.8g, 17mmol) was added in one portion and the whole was refluxed for 2 hours. The reaction mixture was cooled to room temperature and then concentrated in vacuo and purified directly by silica gel column chromatography using a gradient of 8:1 → 4:1 pentane: EtOAc as eluent to give the title compound as a white solid (258, 59%).
1H NMR(CDCl3，400MHz)：δ 2.12(s，3H)，2.34(s，3H)，3.99-4.01(dt，4H)，5.25-5.28(m，4H)，5.34-5.40(d，2H)，6.01-6.10(m，2H)，7.11-7.13(d，2H)，7.24-7.32(m，3H)，7.66(bs，1H)；LRMS m/z(ESCI+)349[MH]+
Preparation 154
4-methyl-3-oxo-pentanoic acid
Ethylisobutyrylacetic acid (21g, 132mmol) was dissolved in 1.5M sodium hydroxide solution (15g in 250ml water) and stirred at room temperature for 16 hours. The solution was cooled to 0 ℃ in an ice bath and then acidified to pH1-2 with 35ml concentrated HCl. The resulting solution was saturated with sodium chloride and then extracted with ethyl acetate (3X 300 ml). The combined extracts were dried over sodium sulfate, filtered, and concentrated in vacuo to give the title compound as a clear oil (16.4g, 95%).
1HNMR(CDCl3400MHz, about 4:1 mixture of keto and enol tautomers) (major keto forms) 1.15-1.16(d, 6H), 2.75-2.71(m, 1H), 3.56(s, 2H).
Preparation 155
4-hydroxy-3-isobutyryl-6-isopropyl-pyran-2-one
4-methyl-3-oxo-pentanoic acid (16.4g, 126mmol) is dissolved in THF (200ml) at room temperature under nitrogen atmosphere and 1, 1-carbonyldiimidazole (22.4g, 138mmol) is added in one portion. The resulting yellow solution was stirred at room temperature for 16 h, then concentrated in vacuo and the residue diluted with DCM (200 ml). The solution was washed with 2N HCl (100ml) and water (100ml) and the aqueous solution was re-extracted with DCM (50 ml). The combined organics were dried over sodium sulfate and then concentrated in vacuo to give the title compound as a yellow oil (11.7g, 80%).
1HNMR(CDCl3，400MHz)δ 1.16-1.18(d，6H)，1.25-1.27(d，6H)，2.71-2.74(m，1H)，3.94-3.97(m，1H)，5.92(s，1H)；LRMS(APCI+)m/z225[MH]+
Preparation 156
4-hydroxy-6-isopropyl-pyran-2-one
4-hydroxy-3-isobutyryl-6-isopropyl-pyran-2-one (11.7g, 52mmol) was dissolved in concentrated sulfuric acid (40ml) and stirred at 130 ℃ for 15 minutes. The obtained black oil was cooled to room temperature and then further cooled to 0 ℃ in an ice bath, and 200ml of crushed ice was added with stirring. The resulting solution was extracted with ethyl acetate (3 × 200ml) and the combined organics were dried over sodium sulfate, filtered and evaporated in vacuo to a light brown oil which was purified by column chromatography using a pentane 3:1 → 30:70 gradient in ethyl acetate as eluent to give the title compound as a light brown oil which solidified on standing (6.1g, 77%).
1HNMR(CDCl3，400MHz)δ 1.20-1.22(d，6H，2.70-2.80(m，1H)，5.58(s，1H)，5.99(s，1H)；LRMS(APCI+)m/z 155[MH]+
Preparation 157
Ethyl-2, 4-diaminobenzyl-6-methyl-3-carboxylate
Ethyl-2, 4-dichloro-6-methyl-3-carboxylate (100mg, 0.43mmol) was dissolved in acetonitrile (2ml) and then treated with triethylamine (2401, 1.70mmol) and benzylamine (1121, 1.02mmol), and the reaction mixture was stirred at 40 ℃ under nitrogen for 18 hours. After cooling to room temperature, the reaction was poured into water and the mixture was extracted with ethyl acetate (3X 5 ml). The combined organics were dried (over MgSO4) And evaporated to give a crude oil, purified with pentane: eluting with ethyl acetate 20:1-5:1, and purifying by silica gel column chromatography. The title compound was obtained as a clear oil (98mg, 61%).
1H-NMR(CDCl3，400MHz)：δ1.21(t，3H)，2.25(s，3H)，4.29(q，2H)，4.40，(d，2H)，4.78，(d，2H)，5.81(s，2H)，7.21-7.42(m，10H)，8.10(brs，1H)，8.30(brs，1H)；LRMSm/z(ESI)376[MH]+
Preparation 158
2, 4-diaminobenzyl-6-methyl-3-carboxylic acid
Ethyl-2, 4-diaminobenzyl-6-methyl-3-carboxylate (40mg, 0.11mmol) was dissolved in methanol (1ml) and treated with 2N sodium hydroxide solution (601, 0.12mmol) and the reaction mixture was stirred at 65 ℃ under nitrogen for 5 h. After cooling to room temperature, the reaction was poured into water, the pH was adjusted to 6-7 using 2N hydrochloric acid, and the mixture was extracted with ethyl acetate (3X 5 ml). The combined organics were dried (over MgSO4) And evaporated to give the title compound as an off-white solid (37mg, quantitative).
1H-NMR(DMSO，400MHz)：δ2.22(s，3H)，4.59，(d，2H)，4.64，(d，2H)，6.20(s，2H)，7.20-7.39(m，10H)；LRMS m/z(ESI)348[MH]+，346[M-H]-
Preparation 159
6-benzylamino-9-benzyl-2-methyl-8-oxo-8, 9-dihydro-7H-purine and 4-benzylamino-9-benzyl-6-methyl-8-oxo-8, 9-dihydro-7H-purine
2, 4-diaminobenzyl-6-methyl-3-carboxylic acid (30mg, 0.09mmol), diphenylphosphonyl azide (25mg, 0.09mmol) and triethylamine (141, 0.10mmol) were mixed in toluene and the reaction was heated at 111 ℃ for 16 hours under nitrogen atmosphere. After cooling to room temperature, the reaction was poured into water and the mixture was extracted with ethyl acetate (3 × 5 ml). The combined organics were dried (over MgSO4) To give a crude oil, which was purified by silica gel column chromatography eluting with pentane: ethyl acetate 1:1 to give the respective title compounds (10mg (i), 11mg (ii), 68% combined yield) as clear oils.
1H-NMR(CDCl3，400MHz)：(I)δ 2.39(s，3H)，4.7，(s，4H)，6.04(s，2H)，7.15-7.39(m，10H)；LRMS m/z(ESI)354[MH]+；(II)δ2.39(s，3H)，4.39，(d，2H)，5.01，(s，2H)，6.19(s，2H)，7.15-7.39(m，10H)。LRMS m/z(ESI)354[MH]+。
Preparation of 160
1-benzyl-2-bromo-1H-imidazole-5-iodo-4-carbonitrile
5-amino-1-benzyl-2-bromo-1H-imidazole-4-carbonitrile (50mg, 0.18mmol) was dissolved in diiodomethane (1ml) and the mixture was heated to 100 ℃. Isoamyl nitrite (971, 0.72mmol) was then added dropwise to the heated reaction mixture using a syringe. Gas evolution was observed. After 30 minutes, the reaction was cooled to room temperature and the solvent was removed under high vacuum. With 100% pentane to 7: 3, pentane: the remaining red crude residue was purified by silica gel column chromatography eluting with ethyl acetate. This gave the title compound (40mg, 60%) as a yellow oil.
1H-NMR(CDCl3，400MHz)：δ 5.22(s，2H)，6.71(s，2H)，7.09(m，2H)，7.28-7.40(m，3H)；LRMS m/z(ESI)388/390[MH]+
Preparation 161
1-benzyl-2-bromo-5- (4-hydroxybut-1-yne) -1H-imidazole-4-carbonitrile (I) and 2- (4-amino-1-benzyl-2-bromo-1H-imidazo [4, 5-c ] pyridin-6-yl) -ethanol (II)
1-benzyl-2-bromo-1H-imidazole-5-iodo-4-carbonitrile (39mg, 0.1mmol) was dissolved in acetonitrile (1ml) and the mixture was purified with triethylamine (201, 0.15mmol), Pd (PhCN)2Cl2(3.8mg, 0.01mmol) and but-1-yn-4-ol (91, 0.12 mmol). The reactants were then placed in a sealed tube at 100 deg.fHeating at deg.C for 2 hr. The reaction was cooled to room temperature and the solvent was removed under vacuum. An ammonia solution in 7N methanol was then added to the remaining crude brown residue and the reaction was heated in a sealed tube at 120 ℃ for 18 hours. The volatile components were then removed under vacuum to yield a crude brown oil. LRMS of this material indicated that the major component was cyclized material (II), m/z (ESI)347/349[ MH]+. This oil was then purified by silica gel column chromatography eluting with 10% pentane in ethyl acetate to give the title compound (I) as a yellow oil (6 mg). Compound (II) could not be recovered from the silica gel column.
1H-NMR(CDCl3，400MHz)：(I)δ 2.75(t，2H)，3.79(t，2H)，5.2(s，2H)，7.18(m，2H)，7.22-7.38(m，3H)；LRMS m/z(ESI)330/332[MH]+
Preparation 162
1-benzyl-5- (-but-4-hydroxy-2-one-1-yl) -1H-imidazole-2-methoxy-4-carbonitrile 1-benzyl-2-bromo-5- (4-hydroxybut-1-yne) -1H-imidazole-4-carbonitrile (6mg, 0.02mmol) was dissolved in methanol (1ml) and the mixture was treated with sodium methoxide (5mg,. times.s). The reaction was then heated at 65 ℃ for 12 hours. The reaction was cooled to room temperature and the solvent was removed under vacuum. A 2N hydrochloric acid solution was then added to the remaining crude residue and the reaction was stirred at room temperature for 2 hours. The volatile components were then removed under vacuum to yield a crude white solid containing predominantly the title compound; LRMS m/z (ESI)300[ MH ]+。
Preparation 163
2, 6-dichloro-4- (N-nitro) amino-pyridines
2, 6-dichloro-4-aminopyridine (1.58g) was dissolved in sulfuric acid (20ml) at 0 ℃ under a nitrogen atmosphere, and nitric acid (2.5ml) was added dropwise. After 30 minutes, the reaction turned orange and was poured slowly into ice water. The precipitate was filtered, washed with water and then dissolved in ethyl acetate. The organic solution was then over MgSO4Dried, filtered and evaporated in vacuo to give the title compound (1.7g) as a yellow solid.
1H NMR(CDCl3，400MHz)δ 7.40(s，2H)，10.4(s，1H)。
Preparation 164
2, 6-dichloro-4-amino-5-nitro-pyridines
2, 6-dichloro-4- (N-nitro) amino-pyridine (1.7g) was dissolved in sulfuric acid (10ml) and heated at 90 ℃ for 30 minutes. The reaction mixture was cooled to room temperature and then poured into ice water to produce a precipitate. The yellow solid was filtered off, collected, dissolved in ethyl acetate and taken up with Na2CO3And (4) washing with an aqueous solution. The organics were then washed with brine, MgSO4Dried, filtered and evaporated in vacuo to give the title compound (1.45g) as a yellow solid.
1H NMR(CDCl3，400MHz)δ 5.70(s，2H)，6.70(s，1H)；LRMS(ES+)m/z 209[MH]+
Preparation 165
2, 6-dibromo-4-amino-5-nitro-pyridines
2, 6-dichloro-4-amino-5-nitro-pyridine (2g) was dissolved in a 33% solution of HBr in acetic acid (20mL) and heated at 90 ℃ for 72 hours in a Teflon lined vessel. The reaction mixture was cooled to room temperature and poured into ice water to produce a precipitate. The resulting solid was filtered off, collected, dissolved in ethyl acetate and then taken up with Na 2CO3And (4) washing with an aqueous solution. The organics were then washed with brine, then MgSO4Dried, filtered and evaporated in vacuo to give the title compound (2g) as a pale yellow solid.
1H NMR(CDCl3，400MHz)δ 5.60(s，2H)，6.90(s，1H)；LRMS(ES+)m/z295，297，299[MH]+
Preparation 166
2, 6-dibromo-4-chloro-5-nitro-pyridine
2, 6-dibromo-4-amino-5-nitro-pyridine (3g) was dissolved in concentrated hydrochloric acid (20ml), and cooled to 0 ℃. Sodium nitrite (3.5g) was added and the reaction mixture was stirred at 0 ℃ for 1 hour. The ice bath was removed and the reaction was warmed to room temperature over 3 hours and then quenched by the addition of ethyl acetate (50ml) and water (100 ml). The organic layer was separated over MgSO4Dried and filtered and evaporated in vacuo to a pale yellow oil which was purified using 35:1 pentane: EtOAc was purified by column chromatography as eluent to give the title compound as a white solid (2.2 g).
1H NMR(CDCl3，400MHz)δ 7.65(s，1H)。
Preparation 167
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine
2, 6-dibromo-4-amino-5-nitro-pyridine (1.53g) was dissolved in THF (20ml), and solid K was added in one portion each2CO3(100mg) and benzylamine (1.1 ml). The reaction mixture was then heated at 70 ℃ for 16 hours. The solvent was removed in vacuo and the crude residue was purified by silica gel column chromatography using 10% ethyl acetate in pentane as eluent to give the title compound (1.2g) as a yellow oil.
1H NMR(CDCl3，400MHz)δ 4.45(d，2H)，4.78(d，2H)，6.20(s，1H)，7.20-7.41(m，10H)，9.41(s，1H)，9.50(s，1H)；LRMS(ES+)m/z 413，415[MH]+
Preparation 168
4, 6-bis-benzylamino-5-nitro-pyridine-2-carboxylic acid methyl ester
N-2, N-4-dibenzyl-6-bromo-3-nitro-pyridine-2, 4-diamine (1g) was dissolved in a mixture of methanol and DMF (2:1, 15ml), followed by addition of triethylamine (0.7ml), triphenylphosphine (30mg) and palladium acetate (27mg), and the mixture was heated at 60 ℃ under 100psi carbon monoxide pressure for 16 h. The reaction mixture was cooled to room temperature, filtered with an Arbocel short plug and evaporated in vacuo to give a yellow residue. This residue was purified by silica gel column chromatography using an 8:1 → 2:1 pentane gradient in ethyl acetate as eluent to give the title compound (0.5g) as a pale yellow solid.
1H NMR(CDCl3，400MHz)δ 3.91(s，2H)，4.58(s，1H)，4.85(d，2H)，6.85(s，1H)，7.05-7.25(m，10H)，9.3(t，1H)，9.55(t，1H)；LRMS(ES+)m/z 393[MH]+
Preparation 169
5-amino-4, 6-benzylamino-pyridine-2-carboxylic acid methyl ester
Methyl 4, 6-bis-benzylamino-5-nitro-pyridine-2-carboxylate (800mg) was dissolved in methanol (30ml), and raney nickel (80mg) was added in one portion, and the mixture was stirred at room temperature under 60psi of hydrogen pressure for 3 hours. The reaction mixture was filtered with an Arbocel short plug and the filtrate was evaporated in vacuo to give the title compound (0.75g) as a yellow residue which was used without further purification; LRMS (ES)+)m/z 363[MH]+
Preparation 170
1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylic acid methyl ester
Methyl 5-amino-4, 6-benzylamino-pyridine-2-carboxylate (0.75g) was dissolved in acetonitrile (40ml), and 1, 1-carbonyldiimidazole (500mg) was added in one portion, and the mixture was heated at 80 ℃ for 6 hours. The reaction mixture was evaporated in vacuo to give a residue, which was purified by silica gel column chromatography using 1:1 pentane in ethyl acetate as eluent to give the title compound as a white solid (100 mg).
1H NMR(d6-DMSO，400MHz)δ 3.75(s，3H)，4.45(d，2H)，4.98(s，2H)，6.48(t，1H)，7.18(s，1H)，7.22-7.47(m，10H)；LRMS(ES+)m/z 389[MH]+
Preparation 171
1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro [4, 5-c ] pyridine-6-carboxylic acid
Methyl 1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylate (0.03g) was dissolved in methanol (1ml) and 1N NaOH solution (2ml), and the mixture was stirred at 40 ℃. After 2 hours, 2ml of 2N HCl was added to precipitate a solid. The solid was filtered off and dried in vacuo to give the title compound as a white solid (25 mg).
1H NMR(CDCl3，400MHz)δ 4.30(s，2H)，4.90(s，2H)，7.10-7.35(m，11H)，10.80(s，1H)；LRMS(ES+)m/z 375[MH]+
Preparation 172
1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylic acid cyclopropylmethylamide
Methyl 1-benzyl-4-benzylamino-2-oxo-2, 3-dihydro-1H-imidazo [4, 5-c ] pyridine-6-carboxylate (0.05g) was dissolved in cyclopropylmethylamine (1ml) and heated in a reaction flask at 80 ℃ for 3 hours. The solvent was removed in vacuo and diethyl ether was added to precipitate the solid. The solid was filtered off and dried in vacuo to yield 40mg of a white solid. This solid was purified by preparative HPLC using a mixture of acetonitrile, water and diethylamine as eluent to give the title compound as a white solid (18 mg).
1H NMR(CDCl3，400MHz)δ 0.22(q，2H)，0.55(q，2H)，1.05(m，1H)，3.22(t，2H)，4.35(d，2H)，4.90(s，2H)，5.90(t，1H)，7.10-7.35(m，11H)，7.85(t，1H)；LRMS(ES+)m/z 428[MH]+
Preparation 173
4-bromo-2-chloro-6-trifluoromethyl-pyridin-3-ylamine
6-trifluoromethyl-pyridin-3-ylamine (150g, 925mmol) was suspended in 500ml acetonitrile. N-chlorosuccinimide (124g, 925mmol) was added to the solution and the mixture was heated at 80 ℃ for 2 hours, then N-bromosuccinimide (165g, 925mmol) was added and the mixture was heated at 80 ℃ for a further 3 hours. The reaction mixture was cooled to ambient temperature, concentrated in vacuo and triturated in 100ml of diethyl ether, and the precipitate was removed by filtration. The resulting filtrate was concentrated in vacuo and purified by silica column chromatography eluting with heptane: EtOAc 4:1 to give the title compound as a dark red oil (220g, 86%).
1H NMR(CDCl3)δ 4.90(bs，2H)，7.67(s，1H)；LRMS(ES)m/z 275/277[MH]+
Preparation 174
N*4*-benzyl-2-chloro-6-trifluoromethyl-pyridine-3, 4-diamine
4-bromo-2-chloro-6-trifluoromethyl-pyridin-3-ylamine (84g, 300mmol) was stirred in 500ml DMSO in the presence of cesium fluoride (46.3g, 305mmol) and benzylamine (66.6ml, 610 mmol). The resulting brown suspension was heated at 150 ℃ for 16 hours. 1500ml of water were added to the cooled suspension and the mixture was extracted with 2X 500ml of diethyl ether. The combined organic extracts were dried (over MgSO4) Concentrated in vacuo and purified by silica column chromatography eluting with heptane: EtOAc 4:1-2:1 to give the title compound as a pale brown solid (15.8g, 17%). The undesired isomer was isolated as a brown oil in the same manner (51.0g, 48%).
1H NMR(CDCl3)δ 3.76(bs，2H)，4.39-4.41(d，2H)，4.53(bs，1H)，6.85(s，1H)，7.34-7.40(m，5H)；LRMS(ES)m/z 302[MH]+。
Preparation 175
4, 6-dihydroxy-2-trifluoromethyl-nicotinic acid ethyl ester
In a three-necked flask, potassium tert-butoxide (5.8g, 51.9mmol) was suspended in 100ml of tetrahydrofuran and a solution of diethyl 1, 3-acetonate (10g, 49.5mmol) in 30ml of tetrahydrofuran was slowly added. Once the addition was complete, the mixture was stirred at room temperature for 30 minutes. In a second three-necked flask equipped with a gas outlet connected to the first, 2, 2-trifluoroacetamide (11.2g, 98.9mmol) was dissolved in 80ml of pyridine, a premixed solution of trifluoroacetic anhydride (20.8g, 98.9mmol) in 30ml of pyridine was slowly added and the resulting gas (2, 2, 2-trifluoromethylacetonitrile) was bubbled directly into the first three-necked flask. Once the addition was complete, the mixture in the second three-necked flask was stirred at room temperature for 30 minutes, then the solvent was removed in vacuo and the residue was poured into 100ml of 4M HCl. The mixture was extracted with 150ml of ethyl acetate. The organic layer was separated, dried over magnesium sulfate and the solvent removed in vacuo. The residue was triturated in dichloromethane and the precipitate was filtered to give 3g of the title compound as a solid.
1H NMR (MeOD): 12.5(s, 1H), 12.4 (broad singlet, 1H), 7.1(s, 1H), 5.05(q, 2H), 2.05(t, 3H); LRMS (ES) +)m/z 252[MH]+
Preparation 176
4, 6-dihydroxy-5-nitro-2-trifluoromethyl-nicotinic acid ethyl ester
4, 6-dihydroxy-2-trifluoromethyl-nicotinic acid ethyl ester (1g, 3.9mmol) was dissolved in 10ml of concentrated sulfuric acid and 2ml of fuming nitric acid was added dropwise at room temperature. Once the addition was complete, the mixture was stirred at room temperature for 30 minutes. The mixture was then poured into crushed ice and the white precipitate collected, dissolved in 50ml ethyl acetate, washed with 50ml water and 50ml brine, dried over magnesium sulfate and the solvent removed in vacuo to yield 1.1g of the title compound as a white solid.
1H NMR(d6 DMSO)：4.25(q，2H)，1.20(t，3H)。
Preparation 177
6-trifluoromethyl-pyridine-2, 4-diol
4, 6-dihydroxy-2-trifluoromethyl-nicotinic acid ethyl ester (15g, 59.7mmol) was dissolved in 250ml concentrated HCl and the mixture was stirred at 115 ℃ for 3 days. The mixture was cooled to 0 ℃ and 0.88 ammonia was added until the pH was-7. The solid formed was filtered, washed with water, azeotroped with toluene, and dried in vacuo to give 9g of the title compound as a white solid.
1H NMR(d6 DMSO)：6.7(s，1H)，6.1(s，1H)；LRMS(ES+)m/z 180[MH]+
Preparation 178
(2-amino-3-nitro-6-vinyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
Mixing (2-amino-6-chloro-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (715mg/1.955mmol), potassium vinyltrifluoroborate (415mg/3.098mmol), [1, 1' -bis (diphenylphosphino) ferrocene ]Dichloropalladium (II) CH2Cl2(90mg/0.11mmol) and triethylamine (0.28ml/2.0mmol) were mixed in iPrOH (8.0ml) and heated at 50 ℃ under N2Heat for 24 hours under atmosphere. Pre-adsorbed directly onto silica and purified by column chromatography to give the title compound as a purple resin (270 mg).
1H NMR (CD3OD, 400MHz) delta 1.15-1.20 (multiplet, 3H), 2.50(s, 3H), 4.05-4.15 (multiplet, 2H), 4.90-4.95 (multiplet, 2H), 5.60(d, 1H), 6.20-6.30 (multiplet, 1H); 6.55-6.65 (multiplet, 2H), 7.20-7.25(d/1H), 7.70 (multiplet/1H), 8.35 (s/1H); LRMS (ES) m/z358[ MH]+。
Preparation 179
(2-amino-6-formyl-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
(2-amino-3-nitro-6-vinyl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (270mg/0.755mmol) was dissolved in acetone (5 ml)/water (5ml), and tetra-ethyl-carbamate was addedOsmium oxide (2.5 wt% overtBuOH) (0.10ml/0.008 mmol). Stirring for 5 min gave a brown solution, then sodium metaperiodate (500mg/3.47mmol) was added and the orange suspension was stirred for 1 h. In EtOAc (100ml) with sodium thiosulfate pentahydrate (20 wt% in H)2O) (50ml) was partitioned. The organics were collected, washed with brine, and Na 2SO4Dried, filtered and concentrated to a brown resin. Purification by column chromatography eluting with EtOAc gave the title compound as a yellow oil (220 mg).
1H NMR (CDCl3, 400MHz) delta 1.15-1.20 (multiplet, 3H), 2.55(s, 3H), 4.10-4.20 (multiplet, 2H), 4.95(s, 2H), 6.06-6.15(br s, 2H), 7.05(s, 1H); 7.10-7.15(d, 1H), 7.60(d/1H), 8.35-8.40(s/1H), 9.75 (s/1H); LRMS (AP) m/z 360[ MH]+。
Preparation 180
(2-amino-3-nitro-6-oxazol-5-yl-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester
(2-amino-6-formyl-3-nitro-pyridin-4-yl) - (6-methyl-pyridin-3-ylmethyl) -carbamic acid ethyl ester (190mg/0.529mmol) was dissolved in MeOH (5 ml). (4-tosyl) methyl isocyanide (124mg/0.634mmol) and anhydrous potassium carbonate (200mg/1.45mmol) were added successively. In N2Stirred under atmosphere for 1 hour and then concentrated in vacuo. In EtOAc (100ml) with H2O (50ml) was dissolved separately. The organics were collected, washed with brine, and Na2SO4Dried, filtered and concentrated to a black resin. Purification by column chromatography eluting with 90:10 DCM/MeOH gave the title compound as a crude brown solid (135 mg); LRMS (ES) m/z399[ MH]+。
Biological data
The ability of compounds of formula (I) and pharmaceutically acceptable salts, solvates and polymorphs thereof to modulate TLR7 receptor activity was confirmed by the PBL/HCV replicon bioassay as detailed below, using the following abbreviations:
EMCV is as follows: encephalomyocarditis virus
IRES: internal ribosome entry site
Huh: huh-7 human hepatoma cell line 7 (parent cell for producing HCV replicon cell line)
luc: luciferase enzyme
ubi, ubi: ubiquitin
neo: neomycin
ET: glutamic acid, threonine (cell culture-adaptive mutation in replicon for assay)
RPMI-FCS: roswell Park clinical Institute (cell culture media of PBL) -fetal bovine serum
PBL: peripheral blood lymphocytes
PBL contains a subpopulation of plasmacytoid dendritic cells that are naturally interferon-producing cells during infection, and thus it is an excellent model for the delineation of interferon inducers. As a very sensitive antiviral bioassay, supernatants from PBLs were tested for antiviral activity in the HCV replicon system. Antiviral EC50Values are defined as the concentration of test compound administered to the PBL after a predetermined amount of PBL medium has been delivered to the cell line containing the HCV replicon, such that the HCV replicon content is reduced by 50%. Although HCV replicon-containing cells respond fully to PBL conditioned media, they do not respond directly to known TLR agonists such as ranisimmod (Resiquimod) and Imiquimod (Imiquimod).
HCV replicon (Huh-5-2[ I389luc-ubi-neo-NS 3-3'/ET ]) Is an in vitro model of HCV replication in which a luciferase reporter is integrated into the HCV sequence and stably retained in the human hepatoma cell line Huh-7. Firefly luciferase reporter molecules are expressed as luciferase-ubiquitin-neomycin phosphotransferase fusion proteins that can be cleaved by host proteases to release luciferase. The replicon also contained an internal EMCV IRES that drove translation of the HCV NS3-5B polymeric protein with cellsAdaptive mutational capacity was cultured to allow high cloning efficiency. Luciferase production has been shown to be directly proportional to the amount of HCV RNA present in the host cell. Firefly luciferase Activity Bright-Glo manufactured by Promega was usedTMDetecting by a luciferase detection system.
Specifically, 1-3mg of test compound is dissolved in 100% (v/v) DMSO to a final concentration of typically 1, 4 or 10mM or above 10mM, depending on the starting concentration desired in the assay. An initial 3-fold serial dilution series of compounds in 100% DMSO was prepared from the stock. The dilution series was further diluted 100-fold with complete RPMI-FCS. Thus, the final concentration of DMSO in the assay was 0.1%, and the final concentration of test compound in the 100% DMSO dilution series was 1/1000.
PBL is 5X 105The cells were inoculated in 90. mu.l/well in an assay plate (TC grade transparent at the bottom of 96 wells) containing the compound prepared above, and incubated for 24 hours.
LucUbisneo HCV replicon cells as 10 cells4Perwell/90. mu.l inoculation. It was incubated for 24 hours. After 24 hours, 10 μ l of the medium was taken from the PBL assay plate and transferred to the HCV replicon assay plate and incubation continued for 48 hours.
| Example No. | 1 | 2 | 3 | 4 | 12 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 |
| EC50(nM) | 933 | 1540 | 266 | 497 | 240 | 252 | 104 | 955 | 107 | 355 | 700 | 215 | 450 |
| Example No. | 23 | 24 | 25 | 26 | 27 | 31 | 32 | 34 | 36 | 37 | 38 | 39 | 40 |
| EC50(nM) | 1080 | 1600 | 1000 | 926 | 453 | 3260 | 1710 | 1300 | 368 | 786 | 848 | 2200 | 427 |
| Example numbering | 49 | 50 | 54 | 55 | 56 | 60 | 61 | 70 | 78 | 82 | 83 | 86 |
| EC50(nM) | 2170 | 1970 | 302 | 2002 | 1300 | 445 | 1680 | 656 | 100 | 127 | 470 | 550 |
Advantageously, the compounds of the invention are selective for the TLR7 receptor rather than one or more other known Toll-like receptors. It is also advantageous that the compounds of the invention have selectivity for the TLR7 receptor rather than one or more cellular kinases and/or one or more purinergic receptors such as adenosine or phosphodiesterase receptors.
Examples 1, 2, 12 and 15 were tested to find selectivity for the TLR7 receptor but not all other known Toll-like receptors.
In addition, examples 1, 2, 12 and 15 were tested to find selectivity for the TLR7 receptor and not for the following targets: MEK (mitogen-activated protein kinase/extracellular signal-regulated kinase), CDK1 (cyclin-dependent kinase-1), CDK2 (cyclin-dependent kinase-2), JNK (stress-activated protein kinase), MSK (mitogen and stress-activated protein kinase), MSK-1, SGK, AMPK, MLCK, CHK-2 and phosphodiesterases PDE3, PDE4 and PDE 5.
In addition, examples 12 and 15 were tested to find selectivity for the TLR7 receptor rather than MAP (mitogen-activated protein kinase).
In addition, example 15 was tested to find selectivity for the TLR7 receptor but not all known adenosine receptors a1, A2a, A2b and A3.
Claims (38)
1. A compound of the formula (I),
or a pharmaceutically acceptable salt or solvate of the compound or tautomer, wherein
(a) Y is a bond, R3Selected from aryl, (C)1-C6) Alkyl and- (C)1-C4) alkylene-O- (C)1-C4) An alkyl group; or
(b) Y is (C)1-4) Alkylene radical, R3Selected from aryl, (C)3-C7) Cycloalkyl and 3-10 membered heterocyclyl;
z is oxygen or absent;
R1selected from H, halogen, OH, CN, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NHSO2R6、-NR6R7、-C(O)R6、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8Aryl and 3-10 membered heterocyclyl;
R2selected from H, halogen, OH, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NR6R7、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8And 3-10 membered heterocyclyl; or
R1And R2Can be connected to form (C)2-C5) An alkylene linkage optionally incorporating 1 or 2 heteroatoms each independently selected from N, O and S;
R5is absent and R4Selected from H, (C)3-C7) Cycloalkyl, aryl, - (CH)2) Aryl, -C (O) R9、-CO2R9、-(C1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9)、-C(O)NR9R10、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2(ii) a Or
R4Is absent and R5Is selected from R9、-C(O)R9、-CO2R9、-(C1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-C(O)NR9R10、-(C1-C6) Aalkyl-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2；
R6And R7Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and- (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R6And R7Together with the nitrogen to which they are attached form a 3-6 membered saturated heterocyclic ring optionally containing another 1 or 2 heteroatoms selected from N, O and S;
R8Is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and phenyl;
R9and R10Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, aryl, - (CH)2) Aryl and 3-10 membered heterocyclyl; or R9And R10Together with the nitrogen to which they are attached form a 3-10 membered heterocyclyl;
R11and R12Independently selected from H and (C)1-C6) An alkyl group; or R11And R12Together with the attached N form a 3-6 membered saturated heterocyclyl, optionally containing another 1 or 2 heteroatoms selected from N, O and S;
the alkyl, cycloalkyl, alkoxy, aryl and heterocyclyl groups are optionally substituted with one or more atoms or groups independently selected from halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12、-(C1-C6) alkylene-NR11R12Aryl and 3-10 membered heterocyclyl;
with the proviso that if R1And R2Is H, and Z and R5If not, then:
(a) when Y-R3When it is ethyl, R4Is not methyl; and is
(b) When Y-R3When it is methyl, R4Not H or methyl.
2. A compound of claim 1, wherein R1Is selected from
(a)H；
(b)CN；
(c) Halogen element
(d) Optionally substituted by 1 to 3 halogen atoms1-C6) An alkyl group;
(e) tetrahydrofuryloxy;
(f) substituted with a 3-6 membered saturated heterocyclic group containing 1-3 heteroatoms independently selected from N, O and S (C) 1-C6) Alkyl, wherein the heterocyclyl is optionally substituted with 1-3 substituents independently selected from CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy and- (C)1-C6) alkylene-O- (C)1-C6) Substituted with a group in alkyl;
(g)-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
(h)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(i) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(j)(C3-C7) A cycloalkyl group;
(k)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(l)-C(O)NR6R7；
(m)-CO2R6；
(n)-C(O)R6；
(o) a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, the 5-and 6-membered aromatic heterocyclic groups being optionally substituted with 1 to 3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(p) phenyl optionally substituted with 1 to 3 halogen atoms;
(q)-NR6R7；
(r)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
wherein
R6、R7、R11And R12As defined in claim 1.
3. A compound according to claim 1 or 2, wherein R is1Is selected from
(a)H；
(b)CN；
(c) Halogen element
(d) Optionally substituted by 1 to 3 halogen atoms1-C6) An alkyl group;
(e) tetrahydrofuryloxy;
(f) substituted by morpholine, piperazine or pyrrolidine optionally substituted by 1 or 2 methyl groups (C) 1-C6) An alkyl group;
(h)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(i) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(j)(C3-C7) A cycloalkyl group;
(k)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(l)-C(O)NR6R7；
(m)-CO2R6；
(n)-C(O)R6；
(o) contains (i)1 to 4 nitrogen atoms, or(ii) (ii) a 5-membered aromatic heterocyclic group of 1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, the 5-and 6-membered aromatic heterocyclic groups being optionally substituted with 1 to 3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(p) phenyl optionally substituted with 1 to 3 halogen atoms;
(q)-NR6R7；
(r)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
wherein the content of the first and second substances,
R6、R7、R11and R12As defined in claim 1.
4. A compound according to any preceding claim, wherein R is1Selected from methyl or ethyl substituted by 1-3 fluorine atoms, cyclopropyl, - (C)1-C2) alkylene-O- (C)1-C2) Alkyl, optionally OH-or cyclopropyl-substituted (C)1-C4) Alkoxy, -COCH3、-CH2OCH3and-CO2CH3。
5. A compound according to any preceding claim, wherein R is1Is cyclopropyl or CF3。
6. A compound according to any one of claims 1 to 3, wherein R1Is a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom, The 5-membered aromatic heterocyclic group is optionally substituted with 1 to 3 atoms or groups independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C3) alkylene-O- (C)1-C4) Alkyl, - (C)1-C4) alkylene-OH, -NR11R12And- (C)1-C3) alkylene-NR11R12Wherein R is11And R12As defined in claim 1.
7. A compound of claim 6, wherein R1Selected from the group consisting of imidazolyl, oxazolyl, oxadiazolyl, triazole, pyrazole and thiazole, each of which is optionally substituted with 1-3 atoms or groups independently selected from the group consisting of halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C3) alkylene-O- (C)1-C4) Alkyl, - (C)1-C4) alkylene-OH and- (C)1-C3) alkylene-NR11R12Wherein R is11And R12As defined in claim 1.
8. A compound of claim 7, wherein R1Selected from unsubstituted oxazolyl, triazole, pyrazole and thiazole.
9. A compound of claim 8, wherein R1Is oxazolyl.
10. A compound according to claim 1, wherein R is2Is selected from
(a)H；
(b) Halogen;
(c) optionally substituted by 1 to 3 halogen atoms1-C6) An alkyl group;
(d) tetrahydrofuryloxy;
(e) is composed of 1-3 of N, OAnd 3-6 membered saturated heterocyclic group substituted with hetero atom in S (C)1-C6) Alkyl, wherein the heterocyclyl is optionally substituted with 1-3 substituents independently selected from CF 3、(C1-C6) Alkyl, (C)1-C6) Alkoxy and- (C)1-C6) alkylene-O- (C)1-C6) Substituted with a group in alkyl;
(f)-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
(g)-(C1-C4) alkylene-N (H) - (C)1-C4) alkylene-O- (C)1-C4) An alkyl group;
(h) optionally substituted by OH or cyclopropyl (C)1-C6) An alkoxy group;
(i)(C3-C7) A cycloalkyl group;
(j)-(C1-C4) Alkylene (C)3-C7) A cycloalkyl group;
(k)-C(O)NR6R7；
(l)-CO2R6；
(m)-C(O)R6；
(n) a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; or a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms, the 5-and 6-membered aromatic heterocyclic groups being optionally substituted with 1 to 3 substituents independently selected from halogen, OH, CF3、(C1-C6) Alkyl, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12And- (C)1-C6) alkylene-NR11R12Is substituted with the atom or group in (1);
(o) phenyl optionally substituted with 1 to 3 halogen atoms;
(p)-NR6R7；
(q)-NH-(C1-C4) alkylene-O- (C)1-C6) An alkyl group;
wherein
R6、R7、R11And R12As defined in claim 1.
11. A compound according to any preceding claim, wherein R is2Is H or methyl.
12. A compound according to any preceding claim, wherein R is2Is H.
13. A compound according to any preceding claim, wherein Z is absent.
14. A compound according to any preceding claim, wherein Y is methylene; and R is 3Selected from aryl; a 5-membered aromatic heterocyclic group containing (i)1 to 4 nitrogen atoms, or (ii)1 to 2 nitrogen atoms and 1 oxygen atom or sulfur atom, or (iii)1 oxygen atom or sulfur atom; and a 6-membered aromatic heterocyclic group containing 1 to 3 nitrogen atoms; the aryl and aromatic heterocycle are optionally substituted with 1-3 atoms or groups independently selected from halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, - (C)1-C6) alkylene-OH, -NR11R12、-(C1-C6) alkylene-NR11R12Aryl and 3-to 10-membered heterocyclic group, wherein R11And R12As defined in claim 1.
15. A compound according to any preceding claim, wherein Y is methylene; and R is3Selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, each of which is optionally substituted with 1-3 substituents independently selected from halogen, (C)1-4) Alkyl, (C)1-C4) Alkoxy and CF3Is substituted with the atom or group in (1).
16. A compound according to any preceding claim, wherein Y is methylene; and R is3Selected from phenyl, pyridin-3-yl and 6-methyl-pyridin-3-yl.
17. A compound according to any preceding claim, wherein R is5Is absent; and is
R4Is selected from- (C)1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-(C1-C6) alkylene-O-C (O) NR 9R10And- (C)1-C6) alkylene-O-P (O) (OH)2Wherein R is9And R10As defined in claim 1.
18. A compound according to any preceding claim, wherein R is4Is H and R5Is absent.
19. The compound of any one of claims 1-16, wherein R4Is absent; and R is5Is selected from- (C)1-C6) alkylene-O-C (O) R9、-(C1-C6) alkylene-O-CO2R9、-(C1-C6) alkylene-O-C (O) NR9R10And- (C)1-C6) alkylene-O-P (O) (OH)2Wherein R is9And R10As defined in claim 1.
20. The compound of claim 1, wherein:
y is methylene;
R1selected from CF3Cyclopropyl and oxazole;
R2is H;
R3selected from phenyl, pyridin-3-yl and 6-methyl-pyridin-3-yl;
R4is H; and is
R5Is absent.
21. A compound of the formula (Ic),
or a tautomer thereof, or a pharmaceutically acceptable salt, solvate, or polymorph of said compound or tautomer,
wherein the content of the first and second substances,
y is methylene;
R1and R2Each independently selected from H, halogen, OH, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, -NR6R7、-CO2R6、-C(O)NR6R7、-C(O)NR6SO2R8Aryl and 3-10 membered heterocyclyl; or
R1And R2Can be connected to form (C)2-C5) An alkylene linkage optionally incorporating 1 or 2 heteroatoms each independently selected from N, O and S;
R3is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, aryl, and 3-10 membered heterocyclyl;
R4Is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10，R5Is absent; or
R5Is selected from R9、-C(O)R9、-CO2R9and-C (O) NR9R10，R4Is absent;
R6and R7Each independently selected from H and (C)1-C6) An alkyl group;
R8is selected from (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl and phenyl;
R9and R10Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, aryl, - (CH)2) Aryl and 3-10 membered heterocyclyl; or
R9And R10Together with the nitrogen to which they are attached form a 3-10 membered heterocyclyl;
the above alkyl, cycloalkyl, alkoxy, aryl and heterocyclyl groups are optionally substituted with one or more groups independently selected from halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy, - (C)1-C6) alkylene-O- (C)1-C6) Alkyl, -NH (C)1-C6) Alkyl, -N ((C)1-C6) Alkyl radical)2Aryl and 3-10 membered heterocyclyl;
with the proviso that if R1And R2Is H, Z and R5If not, then:
(a) when Y-R3When it is ethyl, R4Is not methyl; and is
(b) When Y-R3When it is methyl, R4Not H or methyl.
22. The compound of claim 21Wherein R is1And R2Each independently selected from H, (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl, -CO2H、-CO2(C1-C6) Alkyl and-C (O) NH (C)1-C6) Alkylene (C)3-C7) A cycloalkyl group; or R1And R2Can be connected to form (C)2-C5) An alkylene linkage.
23. The compound of claim 22, wherein R1Selected from H, methyl, n-propyl, isopropyl, cyclopropyl, -CO 2H、-CO2CH3and-C (O) NH (CH)2) A cyclopropyl group; and R is2Selected from H and methyl; or R1And R2Can be connected to form C5-an alkylene linkage.
24. The compound of claims 21-23, wherein R3Is aryl, optionally substituted with one or more groups independently selected from halogen, OH, oxo, CF3、CN、(C1-C6) Alkyl, (C)3-C7) Cycloalkyl group, (C)1-C6) Alkoxy group, (C)1-C6) Alkoxy (C)1-C6) Alkyl, -NH (C)1-C6) Alkyl, -N ((C)1-C6) Alkyl radical)2Aryl and 3-10 membered heterocyclyl.
25. The compound of claim 24, wherein R3Is phenyl, optionally substituted by one or more substituents independently selected from halogen and CF3Substituted with the group (1).
26. A compound of formula (I) according to claim 1, selected from:
4-amino-1-benzyl-6-cyclopropyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one;
4-amino-1-benzyl-6-oxazol-2-yl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one; and
4-amino-1-benzyl-6-trifluoromethyl-1, 3-dihydro-imidazo [4, 5-c ] pyridin-2-one;
or a pharmaceutically acceptable salt or solvate of the compound.
27. A pharmaceutical combination comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate of said compound as claimed in any preceding claim together with one or more pharmaceutically acceptable excipients.
28. The drug combination of claim 27, comprising one or more additional therapeutic agents.
29. The drug combination of claim 28 wherein said one or more additional therapeutically active agents is selected from the group consisting of inhibitors of HCV NS3A protein, HCV NS5A protein, HCV NS4B protein, HCV polymerase, HCV metalloprotease, HCV serine protease, HCV helicase, and p7 protein.
30. A compound of formula (I) as defined in any one of claims 1 to 26 or a pharmaceutically acceptable salt or solvate of said compound for use as a medicament.
31. A compound of formula (I) as defined in any one of claims 1 to 26 or a pharmaceutically acceptable salt or solvate of said compound for use in the treatment of a disease or condition associated with the modulation of the TLR7 receptor.
32. The compound of claim 31, wherein the disease or condition is a viral infection selected from the group consisting of adenovirus, herpesvirus, poxvirus, orthomyxovirus, paramyxovirus, coronavirus, papovavirus, papilloma virus, hepadnavirus, flavivirus, retrovirus, and filovirus.
33. The compound of claim 31, wherein the disease or condition is hepatitis c.
34. Use of a compound of formula (I) as defined in any one of claims 1 to 26 or a pharmaceutically acceptable salt or solvate of the compound in the manufacture of a medicament for the treatment of a disease or condition associated with the modulation of the TLR7 receptor.
35. A method of treating a disease or disorder associated with modulation of TLR7 receptors in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate of the compound as claimed in any one of claims 1 to 26.
36. A process for the preparation of a compound of formula (I),
wherein, in each of the formulae I, XVIII, XVIIa, XIX, XIXa, XXa, XXb, XIV, XV, LIV and LXIII, Y-R3As defined in claim 14, R1As defined in claim 2, R2As defined in claim 10, PG1And PG2Is a nitrogen protecting agent and R13Is (C)1-6) An alkyl group;
the method comprises the following steps:
(a) reacting a compound of formula (XVIII) or (XVIIIa) with a carbonyl donor,
to form the corresponding compound of formula (XIX) or (XIXa),
then deprotecting the compound of formula (XIX) or (XIXa); or
(b) Reducing the compound of formula (XXa),
to form a compound of formula (XXb),
the compound of formula (XXb) is then cyclized by treatment with a protic acid; or
(c) Reducing the compound of formula (XIV),
to form a compound of formula (XV),
then cyclizing the compound of formula (XV) in the presence of a carbonyl moiety; or
(e) Cyclizing the compound of formula (LIV) in the presence of diphenylphosphonyl azide to the corresponding compound of formula (XIXA) as described above, followed by deprotection of the amino protecting group,
Or
(f) (ii) hydrolyzing the compound of formula (LXIII),
37. compounds of the formulae XVIII, XVIIa, XIX, XIXa, XXa, XXb, XIV, XV, LIV and LXIII, where Y-R3As defined in claim 14, R1As defined in claim 2, R2As defined in claim 10, PG1And PG2Is a nitrogen protecting agent and R13Is (C)1-6) An alkyl group.
38. Compounds of the formulae XVIII, XVIIa, XIX, XIXa, XXa, XXb, XIV, XV, LIV and LXIII, where Y-R3As defined in claim 15 or 16, R1R is as defined in any one of claims 4 to 62Is H or methyl, PG1And PG2Is a nitrogen protecting agent and R13Is (C)1-6) An alkyl group.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US60/774,580 | 2006-02-17 | ||
| US60/829,730 | 2006-10-17 | ||
| US60/870,020 | 2006-12-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HK1127920A true HK1127920A (en) | 2009-10-09 |
Family
ID=
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