Patent Application
20060211636
The present invention relates to novel semi-synthetic macrolides having antibacterial activity. More particularly this invention relates to 11,12 y lactone ketolides, to processes for their preparation, to compositions containing them and to their use in medicine.
EP 1114826 inter alia generically discloses macrolide compounds of formula (A) having antibacterial activity
wherein R1 is hydrogen or a hydroxyl protecting group; R4 is inter alia an optionally substituted C1-0 alkyl, X1 is inter alia oxygen, X2 is inter alia CH2, Y is NH, O or S, R5 is inter alia C(O) and R13 is hydrogen or halo.
We have now found novel 11,12 γ lactone ketolides having antibacterial activity.
Thus, the present invention provides compounds of general formula (I)
wherein
R is hydrogen, cyano, (CH2)nA-X—R4 or (CH2)nR5;
R4 is selected from:
R4 is an optionally substituted 9 or 10 membered fused bicyclic heterocyclic having at least one heteroatom selected from oxygen, sulphur or nitrogen;
R5 is a 5 or 6 membered heterocyclic containing at least one nitrogen, optionally substituted by one or two groups selected from oxo or 9 or 10 membered fused bicyclic heterocyclic having at least one heteroatom selected from oxygen, sulphur or nitrogen;
R6 and R7 are independently hydrogen, C1-4 alkyl or phenyl which is optionally substituted by one or two C1-4 alkyl groups;
A further embodiment of the invention provides compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof,
wherein
R is (CH2)nA-X—R4;
A is a group selected from —N(R5)—, —N(R5)C(O)—, —N(R5)S(O)2—, or —N(R5)C(Y)N(R6)—;
R1 is hydrogen, C1-6 alkyl or C3-6 alkenyl;
R2 is hydrogen or a hydroxyl protecting group;
R3 is hydrogen or halogen;
X is optionally substituted C1-10 alkylene chain interrupted by a bivalent radical group selected from —O—, —N(R5)—, —C(O)—, —N(R5)C(Y)N(R6)—, —S(O)m-, —N(R5)C(O)—, —C(O)N(R5)—, —N(R5)C(O)C(O)—, —C(O)O— or —C(NOR7)— or
X is optionally substituted C2-10 alkenylene or optionally substituted C2-10 alkynylene chain wherein said C2-10 alkenylene or C2-10 alkynylene chains are optionally interrupted by a bivalent radical group selected from —O—, —N(R5)—, —C(O)—, —N(R5)C(Y)N(R6)—, —S(O)m-, —N(R5)C(O)—, —C(O)N(R5)—, —N(R5)C(O)C(O)—, —C(O)O— or —C(NOR7)—;
R4 is selected from:
R4 is optionally substituted fused bicyclic heteroaryl groups containing 9 or 10 ring members having at least one heteroatom selected from oxygen, sulphur or nitrogen;
R7 is hydrogen, C1-4 alkyl or phenyl;
Y is an oxygen or a sulphur atom;
n is 0 or an integer from 1 to 5;
m is 0, 1 or 2;
Yet a further embodiment of the invention provides compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof,
wherein
R is hydrogen, cyano or (CH2)nA(CH2)mR4;
R1 is C1-6 alkyl or C3-6 alkenyl;
R2 is hydrogen or a hydroxyl protecting group;
R3 is hydrogen or halogen;
R4 is selected from:
A is a bond or a group selected from N(R5), N[C(O)R5], N(R5)C(O), N(R5)S(O2), N(R5)C(O)O, N═C(R6) or N(R5)C(X)N(R6);
R5 and R6 are independently hydrogen, phenyl, or C1-4 alkyl;
X is an oxygen or a sulphur atom;
n or m are independently 0 or an integer from 1 to 5 with the proviso that the sum of n and m is 0 or an integer from 1 to 5;
and pharmaceutically acceptable salts and solvates thereof.
Suitable pharmaceutically acceptable salts of the compounds of general formula (I) include acid addition salts formed with pharmaceutically acceptable organic or inorganic acids, for example hydrochlorides, hydrobromides, sulphates, alkyl- or arylsulphonates (e.g. methanesulphonates or p-toluenesulphonates), phosphates, acetates, citrates, succinates, tartrates, fumarates and maleates.
The compound of formula (I) and salts thereof may form solvates and the invention includes all such solvates. The solvates may, for example, be hydrates.
References hereinafter to a compound according to the invention include both compounds of formula (I) and their pharmaceutically acceptable acid addition salts together with pharmaceutically acceptable solvates.
In the general formula (I) as drawn, the solid wedge shaped bond indicates that the bond is above the plane of the paper. The broken bond indicates that the bond is below the plane of the paper.
It will be appreciated by those skilled in the art that the compounds of formula (I) when R is not hydrogen contain at least one chiral centre (namely the carbon atom shown as 21 in formula (I)) and this may be represented by the formulae (1a) and (1b).
The configuration for the carbon atom shown as 21 in formula la is hereinafter referred to as the β configuration and in formula 1b as the 21α configuration.
It is to be understood that the two diastereoisomers (1a, 1b) and mixtures thereof are encompassed within the scope of the present invention.
Compounds wherein R2 represents a hydroxyl protecting group are in general intermediates for the preparation of other compounds of formula (I).
When the group OR2 is a protected hydroxyl group this is a non-toxic protecting group, conveniently OR2 is an acyloxy group (i.e. acetoxy or benzyloxy).
The term C1-4 alkyl as used herein as a group or a part of the group refers to a straight or branched alkyl group containing from 1 to 4 carbon atoms; examples of such groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl.
The term C1-10 alkylene chain refers to straight or branched chain containing from 1 to 10 carbon atoms examples of such group include, but are not limited to methylene, ethylene, propylene, isopropylene, n-butylene, isobutylene, tert-butylene, pentylene, n-heptylene, n-octylene, n-nonylene and n-decylene. The term C2-10 alkenylene chain refers to a straight or branched alkylene chain containing from 2 to 12 carbon atoms and having at least one double bond; examples of such groups include ethylene, 2-propenylene, 1-propenylene, isopropenylene, 2-butenylene, 2-pentenylene, 2-hexenylene and the like.
The term C2-10 alkynylene chain refers to a straight or branched alkylene chain containing from 2 to 12 carbon atoms and having at least one triple bond; examples of such groups include ethynylene, 2-propynylene, 1-propynylene, isopropynylene, 2-butynylene, 2-pentynylene, 2-hexenylene and the like.
The term halogen refers to a fluorine, chlorine, bromine or iodine atom.
When R4 is a 5 or 6 membered heteroaryl group according to the invention this includes furanyl, thiophenyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,3-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-triazolyl, 1,3,4-oxadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-oxadiazolyl, 1,2,5-thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,4 oxadiazolyl, 1,2,5-triazinyl or 1,3,5-triazinyl and the like.
The term 9 to 10 membered fused bicyclic heterocyclic group refers to a 5,6/6,5 or 6,6 bicyclic ring system, containing at least one heteroatom selected from oxygen, sulphur or nitrogen, which may be saturated, unsaturated or aromatic. The term 9 to 10 membered fused bicyclic heterocyclic group also refers to a phenyl fused to one 5 or 6 membered heterocyclic group. Example of such groups include benzofuranyl, benzothiophenyl, indolyl, benzoxazolyl, 3H-imidazo[4,5-c]pyridin-yl, dihydrophthazinyl, 1H-imidazo[4,5-c]pyridin-1-yl ,imidazo[4,5-b]pyridyl, 1,3 benzo[1,3]dioxolyl, 2H-chromanyl, isochromanyl, 5-oxo-2,3 dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidyl, 1,3-benzothiazolyl, 1,4,5,6 tetrahydropyridaziyl, 1,2,3,4,7,8 hexahydropteridinyl, 2-thioxo2,3,6,9-tetrahydro-1H-purin-8-yl, 3,7-dihydro-1H-purin-8-yl, 3,4 dihydropyrimidin-1-yl, 2,3 -dihydro- 1,4-benzodioxinyl, benzo[1,3]dioxolyl, 2H-chromenyl, chromanyl, 3,4-dihydrophthalazinyl, 2,3 dihydro-1H-indolyl, 1,3-dihydro-2H-isoindol-2-yl, 2,4,7-trioxo-1,2,3,4,7,8-hexahydropteridinyl, thieno[3,2-d]pyrimidinyl, 4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidinyl, 1,3 dimethyl-6-oxo-2-thioxo-2,3,6,9-tetrahydro-1H-purinyl, 1,2 dihydroisoquinolinyl, 2-oxo-1,3-benzoxazolyl, 2,3-dihydro-5H-1,3-thiazolo [3,2-a]pyrimidinyl, 5,6,7,8-tetrahydro-quinazolinyl, 4-oxochromanyl, 1,3-benzothiazolyl, benzimidazolyl, benzotriazolyl, purinyl, furylpyridyl, thiophenylpyrimidyl, thiophenylpyridyl, pyrrolylpiridyl, oxazolylpyridyl, thiazolylpiridyl, 3,4 dihydropyrimidin-1-yl imidazolylpiridyl, quinoliyl, isoquinolinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pyrazolyl[3,4]pyridine, 1,2 dihydroisoquinolinyl, cinnolinyl, 2,3dihydro-benzo[1,4]dioxin-6-yl, 4,5,6,7-tetrahydro-benzo[b]thiophenyl-2-yl, 1,8naphthyridinyl, 1,6naphthyridinyl, 3,4 dihydro-2H-1,4-benzothiazine, 4,8-Dihydroxy-quinolinyl, 1-oxo-1,2-dihydro-isoquinolinyl or 4-phenyl-[1,2,3]thiadiazolyl and the like.
The term 5 or 6 membered heterocyclic group refers to 5 or 6 ring member containig at least one heteroatom selected from oxygen, sulphur or nitrogen, which may be saturated, unsaturated. Examples of such groups include piperidyl, 2-oxodihydrofuranyl, piperazinyl, morpholinyl, pyrazolidinyl, 1,2 dihydro-3H-pyrazolyl ,imidazolidinyl or pyrrolidinyl and the like.
The term 9 to 10 membered fused bicyclic carbocyclic group refers to a 5,6/6,5 or 6,6 bicyclic carbocyclic ring system which may be saturated, unsaturated or aromatic. It also refers to a phenyl fused to one 5 or 6 membered saturated or unsaturated carbocyclic group. Examples of such groups include naphthyl, 1, 2, 3, 4 tetrahydronaphthyl, indenyl or indanyl and the like.
The term optionally substituted phenyl, optionally substituted 5-6 membered heterocyclic group, optionally substituted 9 to 10 membered fused bicyclic carbocyclic group, optionally substituted 9 to 10 membered fused bicyclic heterocyclic group or optionally substituted 5 or 6 membered heteroaryl group this refers to a 5-6 membered heterocyclic, a 9 to 10 membered fused bicyclic carbocyclic, a 9 to 10 fused bicyclic heterocyclic or 5 or 6 membered heteroaryl as defined above which is substituted by 1 to 4 groups, which may be the same or different, selected from (CH2)pR10 group wherein p is zero or an integer from 1 to 4 and R10 is selected from:
When R4 is an optionally substituted C3-7 cycloalkyl, such a group is optionally substituted by 1 or 2 substituents which may be the same or different and selected from C1-4 alkyl, halogen, cyano, nitro, trifluoromethyl and NR6R7.
When X is a C1-10 alkylene, a C2-10 alkynylene or a C2-10 alkenylene chain which is interrupted by a bivalent radical group selected from —O—, —NR8—, —C(O)—, —N(R8)C(Y)N(R9)—, —S(O)m-, —N(R8)C(O)—, —C(O)N(R8)—, N(R8)C(O)C(O)—, —C(O)O— or —C(NOR6)—, this refers for example to C1-10 alkylene-O—, C1-10alkylene-NR8C(Y)NR9—, C1-10alkylene-NR8—, C1-10 alkylene-C(O)—, C1-10alkylene-S(O)m-, C1-10 alkylene-NR8C(O)—, C1-10 alkylene-C(O)NR8—, C1-10 alkylene-N(R8)C(O)C(O)—, C1-10 alkylene-C(O)O—, C1-10 alkylene C(NOR6), C2-10alkenylene-O—, C2-10alkenylene-NR8—, C2-10alkenylene-C(O)—, C2-10alkenylene-NR8C(Y)NR9—, C2-10 alkenylene-S(O)m-, C2-10 alkenylene-NR8C(O)—, C2-10 alkenylene-C(O)NR8—, C2-10 alkenylene-N(R8)C(O)C(O)—, C2-10 alkenylene-C(O)O—, C2-10 alkenylene-C(NOR6), C2-10alkynylene-O—, C2-10alkynylene-NR8—, C2-10alkynylene-C(O)—, C2-10alkynylene-NR8C(Y)NR9—, C2-10alkynylene-S(O)m-, C2-10alkynylene-NR8C(O)—, C2-10 alkynylene-C(O)NR8—, C2-10alkynylene-N(R8)C(O)C(O)—, C2-10 alkynylene-C(O)O—, C2-10 alkynylene-C(NOR6), or this refers to a C1-10 alkylene, a C2-10 alkenylene or a C2-10 alkynylene chain containing a bivalent radical group selected from:
—O—, —NR8—, —C(O)—, —NR8C(Y)NR9—, —S(O)m-, —NR8C(O)—, —C(O)NR8—.
When A is —N(R6)—, —N(R6)S(O)2— or —N(R6)C(Y)N(R7) and when X is an optionally substituted C1-10 alkylene interrupted by a bivalent radical selected from —O—, —N(R8)—, —N(R8)C(Y)N(R9)—, —S(O)m-, —N(R8)C(O)— or —N(R8)C(O)C(O)— said bivalent radicals are preferably linked to A group by an optionally substituted alkylene chain containing at least two carbon atoms.
When A is —N(R6)—, —N(R6)S(O)2— or —N(R5)C(Y)N(R7) and when X is an optionally substituted C2-10 alkenylene or an optionally substituted C2-10 alkynylene chain and when these chains are interrupted by a bivalent radical selected from —O—, —N(R8)—, —N(R8)C(Y)N(R9)—, —S(O)m-, —N(R8)C(O)— or —N(R8)C(O)C(O)— said bivalent radicals are preferably linked to the A group by an optionally substituted alkenylene or alkynylene chain containing at least 4 carbon atoms and having —CH2— as terminal groups.
A preferred group of compounds of formula (I) are those in which the carbon atom shown as 21 is in the β configuration.
R is preferably (CH2)nA-X—R4 or (CH2)nR5.
R1 is preferably methyl or 2-propenyl.
R2 is preferably hydrogen.
R3 is preferably hydrogen or fluorine.
When R4 is a 5 or 6 membered heteroaryl group this is preferably imidazolyl, imidazolyl, pyrazolyl, thiophenyl, 1,2,3-triazolyl, pyridinyl or furanyl.
When R4 or R5 is a 5 or 6 membered heterocyclic group this is preferably imidazolidinyl or pyrrolidinyl.
When R4 is a 9 or 10 membered fused bicyclic heteroaryl group this is preferably quinolinyl, quinoxalinyl, indolyl, purinyl, 1,3 benzo[1,3]dioxolyl, benzothiazolyl, 1H-benzimidazol-yl 1,3-benzoxazoyl, 1H-pyrrolo[2,3-b]pyridinyl , 1,3-dihydro-2H-isoindolyl, 3H-imidazo[4,5-c]pyridin-3-yl, 3H-imidazo[4,5-b]pyridin-3-yl, 7H-purin-7-yl, 1H-imidazo[4,5-c]pyridin-1-yl, 4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl.
R5 is preferably 1-pyrrolidinyl which is optionally substituted by one oxo or benzo[1,3]dioxolyl.
X is preferably a C1-5 alkylene, a C2-5 alkenylene or a C2-5 alkynylene chain wherein said chains are:
C1-4 alkyl, oxo, C1-4 alkoxy, halogen, cyano, phenoxy, hydroxy, NR8R9, N(R8)C(O)R9, ═NOR6, NR8C(Y)NR9 or optionally substituted phenyl.
n is preferably 0 or 1.
Preferred compounds of the invention are those wherein A is selected from —NH—, —NHC(O)— or —NHC(Y)NH—. Within this class the compounds in which n is 0 or 1 are particular preferred.
A preferred class of compounds of formula (I) are those wherein X is a C1-4 alkylene chain which is optionally interrupted by a bivalent radical selected from —O—, —NH—, —C(O)—, —NHC(O)—, —S(O)2— —S— and /or such a C1-4 alkylene chain is optionally substituted by one group selected from NH2, C1-4 alkyl, oxo or N—OH.
A particularly preferred group of compounds of formula (I) is that wherein R4 is phenyl (optionally substituted by 1 to 3 groups which may be the same or different selected from nitro, amino, methyl, C1-4 alkoxy ie methoxy or hydroxy), 1-imidazolyl (optionally substituted by 1 to 3 groups which may be the same or different selected from pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, phenyl, m-nitrophenyl, dichlorophenyl, C1-4 alkyl i.e methyl, trifluoromethylphenyl, thiophen-2-yl, thiazol-2-yl), 3-trifluoromethylpyrazol-4-yl, 1-pyrazolyl (optionally substituted by 1 to 3 groups which may be the same or different selected from alogen (i.e. chlorine, fluorine), pyridin-2-yl, pyridin-4-yl, quinolin-2-yl, quinolin-4-yl, quinoxalin-2-yl, pyrimidin-4-yl, C1-4 alkyl i.e methyl, 1,3 benzooxazol-2-yl, p-chloro phenyl, difluoro phenyl, pyrazin-2-yl thiazol-5-yl, 1H-indol-3-yl, 1H-indol-2-yl, 3-methoxy-quinoxalin-2-yl, 2-quinolinyl 3-quinolinyl, 4-quinolinyl, 4-pyridinyl, 3-pyridinyl(optionally substituted by one amino), 5methyl furan-2-yl, 3-thiophenyl, 6-methoxy-7H-purin-7-yl, quinoxalin-2-yl, 3-methoxy quinoxalin-2-yl 6-methoxy-2-oxol, 3-benzoxazol-3(2H)-yl, 1H-pyrrolo [2,3-b]pyridin-1-yl, 2-(methylthio)-1H-benzimidazol-1-yl, 1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl, 6-methoxy-2-oxo-1,3-benzoxazol-3 (2H)-yl, 3H-imidazo[4,5-b]pyridin-3-yl, 1,3-benzoxazol-2-yl, benzothiazol-2-yl, 1,3 benzo[1,3]dioxolyl, 3-(5-cyano-3,4-dimethylthien-2-yl)-1H-1,2,4-triazol-1-yl, 2,3-dihydro-benzo[1,4]dioxin-6-yl, 2,4-dimethyl-1,3-thiazol-5-yl or 4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl.
A particularly preferred group of compounds of formula (I) are those wherein R1 is methyl, R2 or R3 is hydrogen, A is —NH—, —NHC(O)—, X is C1-4 alkylene chain which is optionally interrupted by a bivalent radical selected from —O—, —NH—, —C(O)—, —NHC(O)—, —S(O)2— —S— and/or such a C1-4 alkylene chain is optionally substituted by one group selected from NH2, C1-4 alkyl, oxo or N—OH, R4 is a group selected from 4-(pyridin-3-yl)-imidazol-1-yl, 4-(pyridin-3-yl)-imidazol-1-yl, quinoxalin-2-yl, quinoxalin-2-yl, quinolin-4-yl, quinoxalin-2-yl, -(2,3-dihydro-benzo[1,4]dioxin-6-yl, 4-oxo-4,5,6,7-tetrahydro-benzo [b]thiophen-2-yl, 4-methoxy-3-nitro-phenyl, 2-hydroxy-4,5-dimethoxy-phenyl, 3-hydroxy-4-methoxy-phenyl, 3,4-dimethoxy-phenyl, 4-hydroxy-3-methoxy-phenyl, 3-methoxy-quinoxalin-2-yl, 3-amino-4-methoxy-phenyl, 4-(pyridin-3-yl)-imidazol-1-yl, quinolin-4-yl, 4-pyrimidin-4-yl-pyrazol-1-yl, 2-(methylthio)-1H-benzimidazol-1-yl, -[3-(4-chlorophenyl)-1H-pyrazol-5-yl]propylamino)-methylene], 6-methoxy-2-oxo-1,3-benzoxazol-3 (2H)-yl, 1H-pyrrolo [2,3-b]pyridin-1-yl, 3-(2,4-dimethyl-1,3-thiazol-5-yl)-1H-pyrazol-1-yl, 4-phenyl-1-yl, 4-pyridin-4-yl-1H-imidazol-1-yl, thiophen-2-yl, 3-(5-cyano-3,4-dimethylthien-2-yl)-1H-1,2,4-triazol-1-yl, quinolin-3-yl, 1,3-thiazol-2-yl and n is 0 or 1.
Particularly preferred compounds of the invention are selected from:
(11S,21R)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-(pyridin-3-yl)-imidazol-1-yl)-butyramidomethyl)-methylene]-erythromycin A;
(11S,21R)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(pyridin-3-yl)-imidazol-1-yl)-propionamidomethyl)-methylene]-erythromycin A;
(11S,21R)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-ylsulfanyl)-acetamidomethyl)-methylene]-erythromycin A;
(11S,21R)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(quinoxalin-2-ylsulfanyl)-propionamidomethyl)-methylene]-erythromycin A;
(11S,21R)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-[(quinolin-4-ylmethyl)-amino]-methyl)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(quinolin-4-yl)-propionamido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-ylsulfanyl)-acetamido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((4-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-4-oxo)-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-methoxy-3-nitro-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(2-hydroxy-4,5-dimethoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(3-hydroxy-4-methoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(3,4-dimethoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-hydroxy-3-methoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(3-methoxy-quinoxalin-2-ylsulfanyl)-acetamido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-yloxy)-acetamido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(3-amino-4-methoxy-phenyl)-4-oxo-butyramide)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-hydroxymino-4-(4-methoxy-3-nitro-phenyl)-butyramide)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxaline-2-sulfonyl)-acetamide)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(pyridin-3-yl)-imidazol-1-yl)-propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(4-(pyridin-3-yl)-imidazol-1-yl)-ethylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(quinolin-4-yl)-butylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinolin-4-yl)-ethylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-pyrimidin-4-yl-pyrazol-1-yl)-propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-[2-(methylthio)-1H-benzimidazol-1-yl]propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-[3-(4-chlorophenyl)-1H-pyrazol-5-yl]propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(6-methoxy-2-oxo-1,3-benzoxazol-3 (2H)-yl)propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(1H-pyrrolo[2,3-b]pyridin-1-yl)propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-[3-(2,4-dimethyl-1,3-thiazol-5-yl)-1H-pyrazol-1-yl]propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(quinoxalin-2-ylsulfanyl)-propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-phenyl-1H-imidazol-1-yl)propyl)amino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(4-pyridin-4-yl-1H-imidazol-1-yl)ethylamino)-methylene]-erythromycin A; (11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-ylsulfanyl)-ethylamino)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(thiophen-2-yl)-imidazol-1-yl)-propylamino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((3-[4-(5-cyano-3,4-dimethylthien-2-yl)-1H-1,2,4-triazol-1-yl]propyl)amino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((3-quinolin-3-ylpropyl)amino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((3-[4-(3-nitrophenyl)-1H-imidazol-1-yl]propyl)amino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((2-[3-(1,3-thiazol-2-yl)-1H-pyrazol-1-yl]ethyl)amino)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-2-fluoro-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-ylsulfanyl)-acetamido)-methylene]-erythromycin A;
(11S,21R,S)-3-decladinosyl-11,12-dideoxy-2-fluoro-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(pyridin-3-yl)-imidazol-1-yl)-propylamino)-methylene]-erythromycin A.
Further preferred compounds of the invention include:
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(4-(pyridin-3-yl)-imidazol-1-yl)-ethylamino)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinolin-4-yl)-ethylamino)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(pyridin-3-yl)-imidazol-1-yl)-propylamino)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(2-(quinoxalin-2-ylsulfanyl)-acetamido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-((4-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-4-oxo)-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-methoxy-3-nitro-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(2-hydroxy-4,5-dimethoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-oxo-4,5,6,7-tetrahydro-benzo [b]thiophen-2-yl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-quinolin-2-yl-1H-pyrazol-1-yl)propylamino)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(3,4-dimethoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(4-(4-hydroxy-3-methoxy-phenyl)-4-oxo-butyramido)-methylene]-erythromycin A;
(11S,21S)-3-decladinosyl-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl-(3-(4-(thiophen-2-yl)-imidazol-1-yl)-propylamino)-methylene]-erythromycin A.
Compounds according to the invention also exhibit a broad spectrum of antibacterial activity against a wide range of clinical pathogenic microorganisms.
For example, using a standard microtiter broth serial dilution test, compounds of the invention have been found to exhibit useful levels of activity against a wide range of pathogenic microorganisms including Staphylococcus aureus, Streptococcus pneumoniae, Moraxella catarrhalis, Streptococcus pyogenes, Haemophilus influenzae.
Furthermore compounds of the invention are also active against intracellular pathogens such as Chlamydia pneumonia, Clamydia spp, Legionella pneumophila, Mycoplasma pneumonia, species.
The compounds of the invention may therefore be used for treating a variety of diseases caused by pathogenic bacteria in human beings and animals.
Thus, according to another aspect of the present invention, we provide a compound of formula (I) or a physiologically acceptable salt thereof for use in the therapy in a human or animal subject.
According to a further aspect of the invention we provide the use of a compound of formula (I) or a physiologically acceptable salt thereof for the manufacture of a therapeutic agent for the treatment of systemic or topical bacterial infections in a human or animal body.
According to a yet further aspect of the invention we provide a method of treatment of the human or non-human animal body to combat bacterial infections which method comprises administering to the body an effective amount of a compound of formula (I) or a physiologically acceptable salt thereof.
The term treatment is also meant to include prophylaxis.
While it is possible that, for use in therapy, a compound of the invention may be administered as the raw chemical, it is preferable to present the active ingredient as a pharmaceutical formulation.
The compounds of the invention may be formulated for administration in any convenient way for use in human or veterinary medicine and the invention therefore includes within its scope pharmaceutical compositions comprising a compound of the invention adapted for use in human or veterinary medicine. Such compositions may be presented for use in conventional manner with the aid of one or more suitable carriers or excipients. The compositions of the invention include those in a form especially formulated for parenteral, oral, buccal, rectal, topical, implant, ophthalmic, nasal or genito-urinary use.
The compounds according to the invention may be formulated for use in human or veterinary medicine by injection (e.g. by intravenous bolus injection or infusion or via intramuscular, subcutaneous or intrathecal routes) and may be presented in unit dose form, in ampoules, or other unit-dose containers, or in multi-dose containers, if necessary with an added preservative. The compositions for injection may be in the form of suspensions, solutions, or emulsions, in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilising, solubilising and/or dispersing agents. Alternatively the active ingredient may be in sterile powder form for reconstitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.
The compounds of the invention may also be presented for human or veterinary use in a form suitable for oral or buccal administration, for example in the form of solutions, gels, syrups, mouth washes or suspensions, or a dry powder for constitution with water or other suitable vehicle before use, optionally with flavouring and colouring agents. Solid compositions such as tablets, capsules, lozenges, pastilles, pills, boluses, powder, pastes, granules, bullets or premix preparations may also be used. Solid and liquid compositions for oral use may be prepared according to methods well known in the art. Such compositions may also contain one or more pharmaceutically acceptable carriers and excipients which may be in solid or liquid form.
The compounds of the invention may also be administered orally in veterinary medicine in the form of a liquid drench such as a solution, suspension or dispersion of the active ingredient together with a pharmaceutically acceptable carrier or excipient.
The compounds of the invention may also, for example, be formulated as suppositories, e.g. containing conventional suppository bases for use in human or veterinary medicine or as pessaries e.g. containing conventional pessary bases.
The compounds according to the invention may be formulated for topical administration, for use in human and veterinary medicine, in the form of ointments, creams, gels, lotions, shampoos, powders, (including spray powders), pessaries, tampons, sprays, dips, aerosols, drops (e.g. eye ear or nose drops) or pour-ons.
Aerosol sprays are conveniently delivered from pressurised packs, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
For topical administration by inhalation the compounds according to the invention may be delivered for use in human or veterinary medicine via a nebuliser.
The pharmaceutical compositions for topical administration may also contain other active ingredients such as corticosteroids or antifungals as appropriate.
The compositions may contain from 0.01-99% of the active material. For topical administration, for example, the composition will generally contain from 0.01-10%, more preferably 0.01-1% of the active material.
For systemic administration the daily dose as employed for adult human treatment it will range from 2-100 mg/kg body weight, preferably 5-60 mg/kg body weight, which may be administered in 1 to 4 daily doses, for example, depending on the route of administration and the condition of the patient. When the composition comprises dosage units, each unit will preferably contain 200 mg to 1 g of active ingredient.
The duration of treatment will be dictated by the rate of response rather than by arbitrary numbers of days.
Compounds of general formula (I) and salts thereof may be prepared by general method outlined hereinafter. In the following description, the groups R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, n, m, p, q, X, Y and A have the meaning defined for the compounds of formula (1) unless otherwise stated.
Compounds of formula (I), wherein A is —N(R6)C(O)— or a —N(R6)S(O2)—, may be prepared by reaction of compounds of formula (II)
wherein R11 is a cladinose derivative of formula (III), in which R2a is a hydroxy protecting group, or hydroxy, R12 is hydrogen or R12 together R11 is an oxygen atom, with a suitable activated derivative of the acid (IV), HOC(O)XR4 (IV) or with a suitable activated derivative of the sulfonic acid (V) HOS(O)2XR4 (V) respectively and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo; c) removal of the protecting group R2.
Suitable activated derivatives of the carboxyl group or the sulphonic acid include the corrisponding acyl halide, mixed anhydride or activated ester such as a thioester or a pentafluoroester.
The reaction is preferably carried out in a suitable aprotic solvent such as halohydrocarbon (e.g. dichloromethane) or N,N-dimethylformamide optionally in the presence of a tertiary base such as pyridine, dimethylaminopyridine or triethylamine and at a temperature within the range of 0° to 120° C.
Compounds of formula (I) wherein A is —N(R6)C(Y)N(R7)— and R7 is optionally substituted phenyl or C1-4 alkyl, may be prepared from compounds of formula (II), wherein R11 and R12 have the meaning defined above, by reaction with a compound of formula R4XNR7C(Y)L (VI), wherein L is a suitable leaving group as above defined and R7 is phenyl or C1-4 alkyl, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
Compounds of formula (I) wherein A is —N(R6)C(Y)NH— may be prepared from compounds of formula (II)), wherein R11 and R12 have the meaning defined above by reaction with a compounds of formula R4XN═C═Y (VII), if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
Compounds of formula (I) wherein A is —N(R6)— may be prepared from compounds of formula (II)), wherein R11 and R12 have the meaning defined above, by reaction with a compounds of formula R4XL (VIII), wherein L is a suitable leaving group.
If required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
Suitable leaving groups for this reaction include halogen (e.g. chlorine, bromine or iodine) or sulfonyl (e.g. tosyl or methansulfonyl).
Compounds of formula (I) wherein R is A is a N(R6)C(O)O group, in which R6 is hydrogen, phenyl or C1-4 alkyl, may be prepared from compounds of formula (II), wherein R11 and R12 have the meaning defined above, by reaction with the appropriate haloformate compound of formula R4XOC(O)L (IX) wherein L is a suitable leaving group such as halogen (e.g. chlorine or bromine) and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
The reactions of compounds (II) with compounds (VI), (VII), (VIII) or (IX) are conveniently carried out in a solvent such as tetrahydrofuran, acetonitrile or halohydrocarbon (e.g. dichloromethane) optionally in the presence of a base such as triethylamine and at a temperature within the range 0° to 80° C.
Compounds of formula (I) wherein A is a N═C(R6) group, may be prepared from compounds of formula (II), wherein R11 is hydroxy, R12 is hydrogen or R11 together R12 is an oxygen atom, by reaction with a compound of formula R4XCHO (X) and, if required, subjecting the resulting compound to one or more of the following operations a) conversion of the 3-hydroxy group into the 3-oxo and b) removal of the protecting group R2.
The reaction is preferably carried out in a solvent such as a halohydrocarbon e.g. dichloromethane at a temperature within the range 0° to 50° C.
Compounds of formula (I), wherein A is N[C(O)R6] may be prepared by treating a compound of formula (XI) in which R11 and R12 have the meaning as defined for compounds of formula (II), by acylation reaction with the activated carboxylic acid of formula(XIa) R6COOH (XIa).
and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
The reaction is preferably carried out in the presence of a base such as a tertiary amine e.g. triethylamine or pyridine in a solvent such as a halohydrocarbon e.g. dichloromethane at a temperature within the range 0° to 50° C.
Suitable activated derivatives of the carboxyl group or the sulphonic acid include the corrisponding acyl halide, mixed anhydride or activated ester such as a thioester or a pentafluoroester.
Compounds of formula (I) in which R is hydrogen may be prepared by decarboxylation of a compound of formula (XII), wherein R11 and R12 have the meaning as defined for compounds of formula (II), followed, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
The decarboxylation may be carried out in the presence of a lithium salt such as lithium chloride, preferably in an organic solvent such as dimethylsulphoxide.
Compounds of formula (I), wherein R is cyano, may be prepared by cyclisation of chlorine derivatives (XIII) wherein R11 and R12 have the meaning as defined for compounds of formula (II),
with potassium cyanide and conveniently in the presence of a solvent such as a N—N dimethylformamide and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
In a preferred embodiment of the invention, compounds of formula (I) in which A is —N(R6)— and X is C2-10alkylene interrupted by NR8—, —N(R8)C(Y)N(R9)—, —N(R8)C(O)— or —N(R8)C(O)C(O)—, may be prepared by reaction of a compound of formula (XIV),
wherein Xa is C2-10alkyl-N(R8), R11 and R12 are defined as in formula (II), with compounds LXbR4(XV), in which L is a suitable leaving group, Xb is a group selected from C(Y)N(R9), C(Y)N(R9)C1-8alkylene, C(O), C(O)C1-8alkylene, C(O)C(O) or C(O)C1-8 alkylene and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
Compounds of formula (XIV) may be prepared from compounds of formula (II) by reductive N-alkylation with a compound of formula HC(O)C2-9alkyl N(R6)(XVI). The reaction is conveniently carried in a protic solvent such as alcohol, i.e methanol, and in the presence of a suitable metal reducing agent such as sodium borohydride or sodium triacetoxyborohydride.
Compounds of formula (I) in which n is 2 or 3 and wherein X is optionally substituted and/or optionally substituted C1-10 alkylene may be prepared from a phosphite of formula (XVII), wherein R11 and R12 have the meaning defined in formula (II) and R13 is C1-4 alkyl,
by Wittig-Homer reaction with an aldehyde of formula (XVIII), followed by reduction of the corresponding double bond using hydrogen and a metal catalyst (e.g. palladium) and, if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
The Wittig-Homer reaction is carried out in the presence of a suitable organic or inorganic base such as 1,8-diazabicyclo[5.4.0]undec-7-ene or diisopropylethylamine in an aprotic solvent such as dichloromethane, preferably at a temperature ranging between −20° to +80° C.
In the reactions described above cladinose derivatives of formula (III) may be removed by treatment with an organic or inorganic acid. Example of a suitable inorganic acid is hydrochloride. The reaction is carried out in the presence of water or an organic solvent such tetrahydrofuran, dichloromethane or mixture thereof.
In the reactions described above the conversion of the 3-hydroxy group into the 3-oxo may be performed by oxidation reaction using a modified Moffatt-Pfitzner procedure.
Suitable oxidizing agent include N,N-Dimethylaminopropyl-3-ethyl carbodiimide -dimethylsulfoxide. The reaction is suitably carried out in the presence of pyridiniumtrifluoro acetate in a chlorinated solvent such as methylene chloride at −10° C. to 25° C.
In a further embodiment, the oxidation may be carried out using Dess Martin periodinane reagent.
Compounds of formula (I), wherein A is NR6 and in which R6 is optionally substitued C1-4alkyl, may be prepared by treating amino compounds of formula (II), wherein R6 is hydrogen, with an alkylating agent L-R6 (XIX) wherein L is a suitable leaving group in the presence of a base.
Compounds of formula (II) wherein n is 0 may be prepared by intramolecular Michael reaction of compounds of formula (XX) wherein R14 is a suitable nitrogen protecting group, R11 and R12 have the meaning defined in formula (II), in the presence of an organic base such as 1,8-diazabicyclo[5.4.0]undec-7-ene.
The reaction conveniently takes place in an aprotic polar solvent such as acetonitrile, dimethylformamide or an aqueous mixture thereof, followed by removal of the nitrogen protecting group R1-4.
Suitable nitrogen protecting group R14 for use in this reaction includes diarylmethylidene such as diphenylmethylidene.
Compounds of formula (II) wherein n is 1 may be prepared by reduction of a compound of formula (XXI), wherein R11 and R12 have the meaning defined in formula (II).
The reduction may be carried out using conventional reducing agents known in the art for converting a nitrile group into an amino group. Thus for example the reaction may be carried out using hydrogen in the presence of Raney-Nickel as catalyst. The reaction is preferably carried out in an alcoholic solvent such as metyl, ethyl or isopropyl alcohol.
Compounds of formula (XXI) may be prepared by cyclisation of chlorine derivatives (XXII) wherein R11 and R12 have the meaning as defined for compounds of formula (II),
with potassium cyanide and conveniently in the presence of a solvent such as a N—N dimethylformamide, followed if required, subjecting the resulting compound to one or more of the following operations a) hydrolysis of the cladinose derivative (III); b) conversion of the 3-hydroxy group into the 3-oxo and c) removal of the protecting group R2.
Compounds of formula (XVII) may be prepared by heating a compound of formula (XXIII)
in an aprotic solvent such as N,N dimethylformamide at a temperature ranging from 60° to 120° C. in the presence of a base such as 1,8-diazabicyclo[5.4.0]undec-7-ene.
Compounds of formula (XXIII) may be prepared by reaction of chlorine derivatives of formula (XXII) with (R13O)3phosphite. The reaction is carried out in a suitable aprotic solvent such as a hydrocarbon (i.e. toluene or xylene), N,N-dimethylformamide or by neat at a temperature within the range of 80° to 160° C.
Compounds of formula (XXII) may be prepared by reacting the corresponding hydroxy derivatives (XXIV),
wherein R11 and R12 have the meaning defined in formula (II), with a suitable activated derivative of the acid HOCOCH2CI(XXV).
Thus for example the esterification may be carried out by reaction with anhydride (ClCH2CO)2O (XXVI) in a suitable aprotic solvent such as a halohydrocarbon (e.g. dichloromethane) or N,N-dimethylformamide and in the presence of a tertiary base such as pyridine, dimethylaminopyridine or triethylamine and at a temperature within the range of 0° C. to 120° C.
Compounds of formula (XX) may be prepared by treating a compound of formula (XXII) with sodium azide, subjecting the resulting azido compound to the following operations: a) reduction by conventional means for reducing azido group to amino group and b) conversion of the group NH2 into the nitrogen protecting group N═R14 wherein R14 has the meaning defined above and, if required, by removal of the hydroxy protecting group R2. The reduction to amino group may be carried out, for example, in the presence of triphenylphosphine and water.
In a further embodiment of the invention compounds of formula (XX) may be prepared by treating a compound of formula (XXII) with NH4OH in the presence of solvent a suitable solvent for this reaction is dimethylsulphoxide and water.
Compounds of formula (XXIV), may be prepared by reacting 11,12-carbonate erythromycin A derivatives (XXVII), R11 and R12 have the meaning defined in formula (II), with a strong base such as 1,8 diazabicyclo[5.4.0]undec-7-ene.
The elimination reaction may be carried out in an organic solvent such toluene, ethyl acetate, N,N dimethylformamide or a mixture thereof, conveniently with heating. Compounds of formula (XXVII), may be prepared from erythromycin A derivatives of formula (XXVIII),
by conversion of the 2′-hydroxy group into the corresponding hydroxy protected group and by conversion of the 11,12 hydroxy into a carbonate group using triphosgene in a suitable solvent such as dicholorometane, in the presence of pyridine.
Compounds of formula (XXVIII), may be prepared by alkylation of an oxime of formula (XXIX)
wherein R15 is oxime protecting group and R2 and R2a are a hydroxyl protecting group, with a compound of formula L-R1 (XXX) in which L is a suitable leaving group such as a halogen (e.g. chlorine, bromine or iodine) or a sulfonyl (e.g. tosyl, methanesulfonyl), in the presence of a base, followed by hydrolysis of cladinose derivative and conversion of the 3-hydroxy group into the 3-oxo.
The reaction with compound (XXX) is preferably carried out in a solvent such as a halohydrocarbon (e.g. dichloromethane), an ether (e.g. tetrahydrofuran, dimethoxyethane), acetonitrile and the like.
Examples of the bases which may be used include potassium hydroxide, cesium hydroxide, tetraalkylammonium hydroxyde, sodium hydride, potassium hydride and the like, followed by subsequent removal of oxime protecting group.
A suitable oxime protecting goup is R15, for example, 1-isopropoxycyclohex-1-yl.
Oxime compounds (XXIX) may be prepared by reaction of a compound of formula (XXXI) wherein R2 and R2a are hydrogen, using analogous methods to those described in U.S. Pat. No. 6,110,965.
Compounds of formula (I) wherein R3 is halogen may be prepared from compounds of formula (I) in which R3 is hydrogen and R2 is hydroxy protecting group by reaction with a halogenating agent in the presence of an organic or inorganic base.
Suitable halogenating agents include N-fluoro benzensulfonimide, SELECTFLUOR™ for fluorination, pyridinium tribromide or cyanogen bromide for bromination or hexachloroethane for chlorination.
A convenient base for the reaction is selected from sodium hydride, potassium hydride, sodium carbonate, potassium hexamethyldisilazide, lithium diisopropylamide or pyridine.
The reaction is carried out in a solvent such as N,N dimethylformamide, tetrahydrofuran or N-methylpyrrolidone or a mixture thereof, conveniently at a temperature within the range −78° to 60° C.
Alternatively the halo group in position 2 of the macrolide ring may be introduced in an earlier step of the synthesis of compounds of formula (I). Thus, for example, it may be indroduced by treating a compound of formulas (II), (IX), (XII), (XV), (XVI), (XVII), (XVIII), (XIX), (XXII) ) or (XXIII) provided that R11 together with R12 is an oxygen atom, using the method above described for obtaining compound (I) wherein R3 is a halo group.
Compounds of formula(I) wherein R is (CH2)nR5 may be prepared by intramolecolar cyclisation of a compound of formula (XXXII),
wherein L is suitable leaving group such halogen (i.e chlorine or bromine), X is C4-5 alkylene chain optionally substituted by one or two groups selected from oxo 9 or 10 membered fused bicyclic heterocyclic having at least one heteroatom selected from oxygen, sulphur or nitrogen.
The reaction is suitable carried out in the presence of an inorganic base or an organic base. Alternatively compounds of formula (I) wherein R is (CH2)nR5 may be prepared by intramolecular reductive N-alkylation of a compound of formula (XXXIII)
wherein R16 is 9 to 10 membered fused heterocyclic groups, r is 3 or 4. This reaction was carried out conveniently carried in an aprotic solvent such as dichloroethane and in the presence of a suitable metal reducing agent such as sodium borohydride or sodium triacetoxyborohydride.
Compounds of formulas (IV), (V), (VI), (VII), (VIII), (IX), (X), (XIa), (XIII), (XVI), (XVIII), (XIX), (XXI) or (XXV) are known or commercially available compounds or they may be prepared using methods known in the art.
The nitrogen protection reaction may be carried out with an appropriate imine such as benzophenone imine in an aprotic solvent e.g. dichloromethane preferably at room temperature.
Where it is desired to isolate a compound formula (I) as a salt thereof, for example a pharmaceutically acceptable salt, this may be achieved by reacting the compound of formula (I) in the form of the free base with an appropriate amount of suitable acid and in a suitable solvent such as an alcohol (e.g. ethanol or methanol), an ester (e.g. ethyl acetate) or an ether (e.g. diethyl ether or tetrahydrofuran).
Pharmaceutically acceptable salts may also be prepared from other salts, including other pharmaceutically acceptable salts, of the compound of formula (I) using conventional methods.
Suitable hydroxy protecting reagent are those described by T. W. Greene and P. G. M Wuts in Protective Groups in Organic Synthesis 2nd ed., John Wiley & Son, Inc 1991, which is incorporating by reference. Examples of suitable hydroxy protecting reagents include acetic anhydride, benzoic anhydride or a trialkylsilyl chloride in a protic solvent. Examples of aprotic solvent are dichloromethane, NN-dimethylformamide, dimethylsulfoxide, tetrahydrofuran and the like.
The hydroxyl protecting groups may be removed by well known standard procedures. For example when R2a is a trialkyllsilyl group, this may be removed by treatment with tetrabutylammonium fluoride and acetic acid or by reaction with fluoride ions source such as triethyl amine tris (hydrogen fluoride) or this process is conveniently carried out in a solvent such as tetrahydrofuran or acetonitrile. When R2 or R2a is alkanoyl (i.e acetyl or benzoyl) these may be removed by treatment with an alcohol (e.g. methanol or ethanol).
In any of the formulae (I), (II), (XI), (XIV), (XVII), (XXI), (XXXII) or (XXXIII) shown above when there is a an asymmetric carbon atom and no specific configuration is shown then the formula includes all possible configurations.
Specific stereoisomers of the compounds of formula (I) as defined in formula 1a and 1b essentially free of the other stereoisomers may be prepared using general processes described above strarting with the appropriate stereisomer of formula (II).
The process described above for preparing the compounds of formula (II) will in general give a mixture of diastereoisomers.
The individual stereoisomers of the compounds of formula (II) may be separated each other by conventional techniques such as fractional crystallisation or more particularly by column chromatography, using for example a silica column.
In a preferred embodiment of the invention the individual stereoisomer of formula (1a) wherein R is NH2 may be prepared by epimerisation reaction of a compound of formula (1b) or mixture of (1a) and (1b) wherein R is NH2. The reaction is carried out in the presence of benzaldehyde and DBU, followed by hydrolysis of the imine derivative with inorganic acid such as hydrochloride. The reaction is suitable carried out in aprotic solvent such as for example toluene, N—N dimethylformamide.
The assignment of the R or S configuration at the 21-position have been made according to the rules of Cahn, Ingold and Prelog, Experientia 1956, 12, 81.
When examples are obtained as a diastereoisomeric mixture of 21R and 21S, unless otherwise stated, the 1H-NMR spectra refers to the 1H-NMR spectra of the predominant diastereoisomer (i.e. 21 S).
In the Intermediates and Examples unless otherwise stated:
Proton Magnetic Resonance (1H-NMR) spectra were recorded at 500 MHz, chemical shifts are reported in ppm downfield (6) from Me4Si, used as internal standard, and are assigned as singlets (s), doublets (d), doublets of doublets (dd), triplets (t), quartets (q) or multiplets (m). Mass spectra were acquired with a Hewlett Packard 1100 MSD system equipped with a binary pump (Agilent Technologies), operating in positive electrospray ionisation mode. LC/MS (Liquid Chromatography/Mass Spectroscopy) data were obtained by using a HP 1100 LC system (Agilent Technologies) equipped with a Sedex Evaporative Light Scattering Detector model 75 (Sedere) coupled with a Platform LCZ Mass Spectometer (Micromass) operating in positive electrospray ionisation mode. The chromatographic analysis conditions were: column Waters XTerra MS C18 (4.6×30 mm, 2.5 μm); flow rate 0.8 ml/min; mobile phase: aqueous solution of NH4OAc (10 mM, pH 6.8) (A) and acetonitrile (B).
LC (Liquid Chromatography) purifications were performed with a Waters 600 semi-preparative system equipped with a binary pumping system and a Jasco-UV detector. The chromatographic analysis conditions were: column Supelcosil ABZ+Plus (10 cm×21.2 mm, 5 μm); flow rate 8 ml/min; mobile phase: aqueous solution of NH4OAc (10 mM, pH 6.8) (A) and acetonitrile (B).
Column chromathography was carried out over silica gel 60 (230-400 mesh ASTM—Merck AG Darmstaadt, Germany). The TLC (Thin Layer Chromatography) monitoring was performed using Merck 60 F254 as TLC plate.
Phase separations were done by using Microfiltration Device—Filter Tube with polypropylene support (Whatman).
Resin washings were carried out on Extract-clean Tube (Alltech).
Purifications of crude products were performed by SCX-cartridges (Varian).
PS-Trisamina resin (polystyrene based) (Argonaut Technologies Inc.) was used to remove the excess of reagents.
Abbreviations which have been used in the description of the synthetic methods that follow are: Brine for aqueous saturated solution of sodium chloride, DBU 1,8-diazabicyclo[5.4.0]undec-7-ene, DCE for 1,2-dichloroethane, DCM for dichloromethane, DIPEA for N,N-diisopropylethylamine, DMAP for 4-dimethylaminopyridine, DMF for N,N-dimethylformamide, DMSO for methyl sulfoxide, EDC for 1-(3-dimethylaminopropyl-3-ethylcarbodiimide hydrochloride, Et2O for diethyl ether, EtOAc for ethyl acetate, HATU for O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate, HOBT for 1-hydroxybenzotriazole hydrate, iPrOH for 2-propanol, MeOH for methanol, MTBE for tert-butyl methyl ether, TEA for triethylamine and THF for tetrahydrofuran, wt for weight.
To a solution of 2′-O-acetyl-3-decladinosyl-6-O-methyl-3-oxo-erythromycin A (0.500 g) in anhydrous DCM (20 mL) under nitrogen atmosphere, pyridine (1.5 mL) and phosgene (20% sol. in toluene, 1 mL) were sequentially added. The reaction mixture was stirred overnight at room temperature then quenched with a saturated NaHCO3 aqueous solution (50 mL). The organic phase was washed with water (50 mL), dried over Na2SO4, concentrated under reduced pressure and the crude product purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.360 g).
TLC: DCM\MeOH 90\10 (Rf=0.6).
To a solution of intermediate 1 (0.210 g) in 2\1 mixture of EtOAc\toluene (6 mL), DBU (0.05 mL) was added and the mixture was heated to 85° C. for 6 h. The reaction mixture was allowed to reach room temperature, the solvent evaporated and the crude product purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.150 g).
TLC: DCM\MeOH 90\10 (Rf=0.7).
To a solution of intermediate 2 (0.150 g) in anhydrous DCM (3 mL) cooled to 0° C., pyridine (0.05 mL), chloroacetic anhydride (0.065 g) and DMAP (5 mg) were sequentially added under nitrogen atmosphere. The reaction mixture was stirred for 4 h then quenched with water (10 mL) and extracted with DCM (2×10 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 80\20) to give the title compound (0.060 g).
TLC: DCM\MeOH 90\10 (Rf=0.8).
To a solution of example 1 (0.137 g) in iPrOH (20 mL) Raney-Nickel (0.100 g) was added. The reaction mixture was saturated with hydrogen (5 atm) and stirred at room temperature for 24 h. After removing the catalyst by filtration and evaporating the solvent under reduced pressure, the crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.012 g).
1H-NMR (CDCl3) δ: 7.61, 7.07 (t+t, 1H), 4.94, 4.82 (dd+dd, 1H), 4.76 (m, 1H), 4.39, 4.38 (d+d, 1H), 4.24, 4.22 (d+d, 1H), 3.83 (q, 1H), 3.55 (m, 1H), 3.26 (m, 1H), 3.13 (m, 1H), 3.02, 2.90 (d+d, 1H), 2.96 (m, 1H), 2.70 (m, 1H), 2.65, 2.64 (s+s, 3H), 2.60, 2.50 (m+m, 1H), 2.26 (s, 6H), 2.07, 2.06 (s+s, 3H), 1.94 (m, 1H), 1.77-1.64 (m, 2H), 1.55 (m, 1H), 1.50, 1.46 (s+s, 3H), 1.33 (d+d, 6H), 1.32 (s, 3H), 1.21 (d, 3H), 1.17 (m, 1H), 1.14 (s, 3H), 0.97, 0.96 (d+d, 6H), 0.83, 0.80 (t+t, 3H).
TLC: DCM\MeOH 95\5 (Rf=0.65).
A solution of intermediate 4 (0.010 g) in MeOH (1 mL) was stirred for 48 h, then concentrated under reduced pressure to give the title compound (0.009 g).
1H-NMR (CDCl3) δ: 7.61, 7.09 (t+t, 1H), 4.94, 4.83 (dd +dd, 1H), 4.86 (bm, 1H), 4.32, 4.30 (d+d, 1H), 4.26, 4.24 (d+d, 1H), 3.86 (m, 1H), 3.56 (m, 1H), 3.13-3.10, 2.96 (m, 3H) 3.02 (m, 1H), 2.88 (bs, 1H), 2.66 (s, 3H), 2.50 (m, 1H), 2.30 (s, 6H), 1.95 (m, 1H), 1.85, 176 (m, 1H), 1.70-1.60 (m, 2H), 1.55 (m, 1H), 1.49, 1.46 (s+s, 3H), 1.4-1.3 (m, 6H), 1.30-1.14 (m, 2H), 1.0-0.9 (d, 3H), 0.84, 0.81 (t+t, 3H).
TLC: DCM\MeOH 95\5 (Rf=0.45).
To a solution of 6-O-methyl-erytrhromycin A (50 g) in anhydrous DCM (240 mL), TEA (26.1 mL), DMAP (0.392 g) and acetic anhydride (15.2 mL) were added at 0° C. under nitrogen atmosphere. The resulting mixture was stirred at 0° C. for 45min and overnight at room temperature. The mixture was then diluted with a saturated NH4Cl aqueous solution (240 mL) and extracted with DCM (2×200 mL). The aqueous phase was neutralised with a saturated NaHCO3 aqueous solution and extracted again with DCM (2×200 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to give the title compound (50.7 g).
m\z ([MH]+)=832.
To a solution of intermediate 6 (200 g) in anhydrous DCM (1600 mL) cooled to 0° C., pyridine (117 mL) and a solution of triphosgene (71.2 g) in anhydrous DCM (400 mL) were added under nitrogen atmosphere. The resulting mixture was stirred at 0° C. for 30 min and then at room temperature for 15 h. The mixture was then diluted with water (750 mL) and extracted with DCM (2×500 mL). The organic layer was washed with water (3×300 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (200 g).
m\z ([MH]+)=858.
To a solution of intermediate 7 (50.5 g) in a 2\1 mixture of toluene\EtOAc (675 mL), DBU (12 mL) was added at room temperature. The resulting mixture was stirred at 85° C. for 8 h and at room temperature for 5 h. The reaction mixture was then diluted with brine (250 mL), extracted with EtOAc (2×350 mL) and dried over Na2SO4. The solvent was evaporated under reduced pressure and the crude material purified by crystallisation (from acetone\water) to give the title compound (46 g).
m\z ([MH]+)=814.
To a solution of intermediate 8 (20 g) in anhydrous DCM (340 mL) cooled to 0° C. pyridine (6 mL) and chloroacetic anhydride (8.4 g) were added under nitrogen atmosphere and the reaction was allowed to reach room temperature. After 16 h the reaction mixture was washed with water (300 mL), a saturated NH4Cl aqueous solution (150 mL) and brine (150 mL) then extracted with DCM (2×300 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was dissolved in acetone (50 mL) and water (100 mL) was added under vigorous stirring. The precipitate was filtered and dried in vacuo to give the title compound (20.4 g).
m\z ([MH]+)=890.
To a solution of intermediate 9 (20.2 g) in THF (200 mL) cooled to 0° C. a 3N HCl aqueous solution (400 mL) was added dropwise. The reaction mixture was allowed to reach room temperature and stirred overnight. The solution was neutralised with a saturated NaHCO3 aqueous solution and extracted with DCM (2×200 mL). The organic layer was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by quick filtration on silica gel (eluting with: DCM\MeOH 95\5) to give the title compound (15.4 g).
m\z ([MH]+)=690.
To a solution of intermediate 10 (0.400 g) in anhydrous DMF (30 mL) potassium cyanide (0.380 g) was added under nitrogen atmosphere. The mixture was stirred at room temperature for 2 h, quenched with a saturated NaHCO3 aqueous solution (50 mL) and extracted with DCM (70 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.430 g).
TLC: DCM\MeOH 90\10 (Rf=0.50).
To a solution of intermediate 11 (0.100 g) in iPrOH (10 mL) Raney-Nickel (0.100 g) was added. The mixture was saturated with hydrogen (5 atm) and stirred at room temperature for 24 h. After removing the catalyst by filtration and evaporating the solvent, the crude product was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound 12 (0.038 g) and the title compound 13 (0.027 g).
TLC: DCM\MeOH 90\10, Rf (intermediate 12)=0.43
TLC: DCM\MeOH 90\10, Rf (intermediate 13)=0.2
To a solution of intermediate 13 (0.054 g) in anhydrous DCM (2 mL) pyridine (0.010 mL), chloroacetic anhydride (0.016 g) and DMAP (catalytic amount) were sequentially added under nitrogen atmosphere. The mixture was stirred at 0° C. for 1 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (5 mL) and extracted with DCM (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 98\2) to give the title compound (0.036 g).
TLC: DCM\MeOH 90\10 (Rf=0.43).
To a solution of intermediate 13 (0.100 g) in anhydrous DCM (4 mL) pyridine (0.024 mL), 4-chlorobutyryl chloride (0.016 mL) and DMAP (catalytic amount) were sequentially added under nitrogen atmosphere. The mixture was stirred at 0° C. for 2 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (20 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.100 g).
TLC: DCM\MeOH 90\10 (Rf=0.40).
m\z ([MH]′)=789.
To a solution of intermediate 10 (3 g) in DMSO (40 mL) a 32% aqueous ammonia solution (8 mL) was added dropwise over 10 min at room temperature. The reaction mixture was stirred for 2 h at 50° C. After cooling to 0° C., water (40 mL) was added and the mixture was extracted with MTBE (2×45 mL). The collected organic layers were washed with brine (40 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (2.5 g).
TLC: DCM\MeOH 10\1 (Rf=0.28).
A solution of intermediate 16 (4.0 g) and benzophenone imine (2.6 mL) in anhydrous DCM (40 mL) was stirred at room temperature under nitrogen atmosphere. After 36 h the reaction was quenched with water (100 mL) and extracted with DCM (3×300 mL). The organic layer was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (3.5 g).
TLC: DCM\MeOH 10\1 (Rf=0.38).
A solution of intermediate 17 (3.0 g) and DBU (0.540 mL) in acetonitrile (135 mL) and water (15 mL) was stirred at room temperature for 3 h. After evaporating the solvent, the crude material was dissolved in DCM (300 mL) and washed with water (100 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (3.0 g).
TLC: DCM\MeOH 10\1 (Rf=0.38).
To a solution of intermediate 8 (0.500 g) in anhydrous toluene (100 mL) under nitrogen atmosphere, pyridine (0.250 mL) and methylmalonyl chloride (0.158 mL) were added at 0° C. The temperature was allowed to reach room temperature. After stirring for 1 h, water (50 mL) was added, the organic layer was washed with brine (50 mL) and dried over Na2SO4. The solvent was evaporated under reduced pressure and the crude material purified by quick filtration on silica gel to give the title compound (0.560 g).
m\z ([MH]+)=914.
A solution of intermediate 19 (0.500 g) in a 2N HCl aqueous solution (50 mL) and THF (1 mL) was stirred at room temperature for 6 h. Then, the mixture was cooled to 0° C. and a saturated potassium carbonate aqueous solution was added until pH=9 was obtained. The mixture was extracted with DCM (2×50 mL), the organic phase washed with brine (25 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH=95\5) to give the title compound 20 (0.180 g), and the title compound 21 (0.180 g).
m\z ([MH]+) (intermediate 20)=714.
m\z ([MH]+) (intermediate 21)=700.
A solution of intermediate 20 (0.150 g) and DBU (0.050 mL) in water (1.5 mL) and acetonitrile (13.5 mL) was stirred at 40° C. for 6 h. After evaporating the solvent under reduced pressure, the residue was dissolved in DCM (20 mL), the organic phase washed with water (50 mL), dried over Na2SO4 and concentrated under reduce pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.070 g).
m\z ([MH]′)=714.
A mixture of intermediate 22 (0.050 g) and lithium chloride (0.006 g) in DMF (1 mL) was heated to reflux for 4 h. The mixture was allowed to reach room temperature and poured into a 3% NaHCO3 aqueous solution at 0° C., then extracted with DCM (2×15 mL). The organic phase was washed with water (2×10 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (DCM\MeOH: 95\5) to give the title compound (0.010 g).
m\z ([MH]+)=656.
To a solution of example 7 (0.335 g) in anhydrous acetonitrile (3 mL) chloroacetaldehyde (50 wt % solution in water, 0.127 mL) was added under a nitrogen atmosphere and the resulting mixture was stirred for 20 h at room temperature. Sodium cyanoborohydride (1M in THF, 0.500 mL) and acetic acid (0.041 mL) were added and the reaction mixture was stirred for 6 h at room temperature. After evaporating the solvent the residue was dissolved in DCM (25 mL) and washed with a 5% NaHCO3 aqueous solution (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.300 g).
m\z ([MH]+)=731.
To a solution of intermediate 13 (0.028 g) in anhydrous DMF (2.5 mL) 3-quinolinecarboxylic acid (0.008 g), HATU (0.017 g) and DIPEA (0.017 mL) were sequentially added under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 h and then the solvent evaporated under vacuum. The residue was dissolved in DCM (15 mL) and washed with water (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash chromatography to give the title compound (0.022 g).
TLC: DCM\MeOH 90\10 (Rf=0.37).
To a solution of intermediate 13 (0.050 g) in anhydrous DMF (5 mL) intermediate 49 (0.019 g), HATU (0.030 g) and DIPEA (0.031 mL) were sequentially added under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 3 h, and then the solvent was removed under vacuum. The residue was dissolved in DCM (20 mL), the organic phase washed with water (15 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography to give the title compound (0.042 g).
TLC: DCM\MeOH 90\10 (Rf=0.37).
To a solution of intermediate 13 (0.050 g) in anhydrous DMF (5 mL) intermediate 48 (0.020 g), HATU (0.030 g) and DIPEA (0.031 mL) were sequentially added under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 h and then the solvent was removed under vacuum. The residue was dissolved in DCM (15 mL), the organic phase washed with water (10 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash chromatography to give the title compound (0.042 g).
TLC: DCM\MeOH 90\10 (Rf=0.28).
To a solution of 3-(1H-imidazol-4-yl)-pyridine (0.005 g) in anhydrous DMF (2 mL) cooled to 0° C. sodium hydride (0.001 g) was added under nitrogen atmosphere. The reaction mixture was stirred at 0° C. for 1 h then a solution of intermediate 14 (0.030 g) in anhydrous DMF (1 mL) was added. After 16 h at room temperature the reaction was quenched with water (5 mL) and the mixture extracted with DCM (10 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by preparative TLC (eluting with: DCM\MeOH 90\10) to give the title compound (0.020 g).
TLC: DCM\MeOH 90\10 (Rf=0.35).
To a solution of intermediate 15 (0.095 g) in anhydrous DMF (8 mL) cooled to 0° C., sodium hydride (0.004 g) was added under nitrogen atmosphere. The mixture was stirred at 0° C. for 1 h and at room temperature for 18 h. Water (5 mL) was added and extracted with a mixture Et2O\DCM 80\20 (3×10 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.050 g).
m\z ([MH]+)=753.
To a solution of intermediate 13 (0.075 g) in anhydrous DMF (7 mL) (quinoxalin-2-ylsulfanyl)-acetic acid (0.029 g), HATU (0.046 g) and DIPEA (0.030 mL) were sequentially added under nitrogen atmosphere. The mixture was stirred at room temperature for 6 h. The solvent was evaporated under reduced pressure and the residue dissolved in DCM (20 mL). The organic phase was washed with a saturated NaHCO3 aqueous solution (2×15 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.069 g).
m\z ([MH]+)=887.
To a solution of intermediate 13 (0.030 g) in anhydrous DMF (3 mL) 3-(quinoxalin-2-ylsulfanyl)-propionic acid (0.012 g), HATU (0.018 g) and DIPEA (0.012 mL) were sequentially added under nitrogen atmosphere. The mixture was stirred at room temperature for 6 h then the solvent was evaporated under reduced pressure. The residue was dissolved in DCM (15 mL), the organic phase washed with a saturated NaHCO3 aqueous solution (2×10 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.035 g).
m\z ([MH]+)=901.
A solution of intermediate 13 (0.090 g) and quinoline-4-carbaldehyde (0.023 g) in anhydrous toluene (5 mL) was heated at 50° C. for 8 h under nitrogen atmosphere. Solvent evaporation under reduced pressure gave the title compound (0.100 g).
m\z ([MH]+)=824.
To a solution of 6-OAllyl erythromycin A (1 g) in anhydrous DCM (5 mL) cooled to 0° C., TEA (0.5 mL), DMAP (0.008 g) and acetic anhydride (0.31 mL) were added under nitrogen atmosphere. The resulting mixture was stirred at 0° C. for 1 h and overnight at room temperature. Then a saturated NH4Cl aqueous solution (30 mL) was added the mixture extracted with DCM (2×50 mL). The aqueous phase was neutralised with a saturated NaHCO3 aqueous solution and extracted again with DCM (2×50 mL). The combined organic layers were dried over Na2SO4 and evaporated under reduced pressure to give the title compound (1 g).
m\z ([MH]+)=858.
To a solution of intermediate 33 (4.13 g) in anhydrous DCM (85 mL), pyridine (0.8 mL) and then phosgene (20% sol in toluene, 2.55 mL) were added at 0° C. under nitrogen atmosphere. The resulting mixture was stirred at 0° C. for 30min and then at room temperature for 1 h. The reaction mixture was then diluted with water (150 mL) and extracted with DCM (2×200 mL). The organic layer was washed with water (3×100 mL), dried over Na2SO4 and evaporated under reduced pressure to give the title compound (4.02 g).
m\z ([MH]+)=884.
To a solution of intermediate 34 (4.02 g) in toluene (45 mL) and EtOAc (23 mL), DBU (0.71 mL) was added at room temperature. The resulting mixture was heated to 85° C. for 6 h. The mixture was then diluted with brine (100 mL), extracted with EtOAc (2×200 mL) and dried over Na2SO4. Solvent evaporation under reduced pressure and purification by flash chromatography of the crude material (eluting with: DCM\MeOH\NH4OH 954\0.01) gave the title compound (1.70 g).
m\z ([MH]+)=840.
To a solution of intermediate 35 (1.7 g) in anhydrous DCM (50 mL) cooled at 0° C., pyridine (0.66 mL), DMAP (0.012 g) and chloroacetic anhydride (0.695 g) were added under nitrogen atmosphere. The resulting mixture was stirred for 30 min at 0° C. and then at room temperature for 2.5 h. The mixture was diluted with water (50 mL), neutralised with a saturated aqueous solution of NaHCO3 and extracted with DCM (2×100 mL). The organic phase was washed with water (3×75 mL), dried over Na2SO4 and evaporated under reduced pressure to give the title compound (1.8 g).
m\z ([MH]+)=916.
To a solution of intermediate 36 (1.8 g) in THF (35 mL), a 6N HCl aqueous solution (10 mL) was added at 0° C. The resulting mixture was stirred overnight at room temperature, then it was diluted with water (50 mL). The pH of the solution was brought to 8-9 by addition of solid NaHCO3 and a 1% NaOH aqueous solution, then the aqueous phase was extracted with DCM (2×100 mL). Solvent evaporation under reduced pressure and treatment of the residue with Et2O gave the title compound (1.4 g).
m\z ([MH]+)=716.
To a solution of intermediate 37 (1.3 g) in anhydrous DMF (40 mL) potassium cyanide (0.500 g) was added under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 1 h, quenched with a 5% NaHCO3 aqueous solution (50 mL) and extracted with DCM (2×50 mL). The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH 95\4\0.01) to give the title compound (0.42 g).
m\z ([MH]+)=707.
To a solution of intermediate 37 (1.42 g) in anhydrous DMF (10 mL), sodium azide (0.211 g) was added under nitrogen atmosphere. The mixture was heated to 80° C. for 10 min then quenched with water (100 mL) and extracted with EtOAc (3×200 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (1.36 g).
m\z ([MH]+)=723.
To a solution of intermediate 39 (1.36 g) in THF (25 mL), triphenylphosphine (0.985 g) and water (0.034 mL) were added. The mixture was stirred at room temperature overnight. After evaporating the solvent, the residue was dissolved in DCM (100 mL) and the solution washed with water (2×100). The organic layer was dried over Na2SO4 and concentrated under vacuum to give the title compound (1.30 g).
m\z ([MH]+)=697.
A solution of intermediate 40 (1.30 g) and benzophenone imine (0.9 mL) in anhydrous DCM (15 mL) was stirred at room temperature under nitrogen atmosphere. After 30 h the reaction was quenched with water (50 mL) and extracted with DCM (3×100 mL). The organic layer was dried over Na2SO4, filtered and concentrated under vacuum to give the title compound (1.60 g).
m\z ([MH]+)=861.
A solution of intermediate 41 (1.60 g) and DBU (0.3 mL) in acetonitrile (90 mL) and water (9 mL) was stirred at room temperature for 2 h. Then the solvents were evaporated and the crude material was dissolved in DCM (100 mL). The mixture was washed with water (2×100 mL), the organic phase dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH3 9.5\0.4\0.03) to give the title compound (0.528 g).
m\z ([MH]+)=861.
To a stirred suspension of sodium hydride (0.769 g) in anhydrous DMF (30 mL) under nitrogen atmosphere, a solution of 3-(1H-imidazol-4-yl)-pyridine (3 g) in anhydrous DMF (10 mL) was added dropwise at room temperature. The mixture was stirred for 30 min then methyl bromoacetate (2.4 mL) was added dropwise. After stirring for 2 h the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (200 mL) and washed with water (50 mL). The aqueous phase was further extracted with EtOAc (2×40 mL). The organic layer was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (2.78 g).
m\z ([MH]+)=218.
To a stirred suspension of sodium hydride (0.165 g) in anhydrous DMF (2.5 mL) cooled at 0° C. a solution of 3-(1H-imidazol-4-yl)-pyridine (1 g) in anhydrous DMF (5 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of methyl 3-bromopropanoate (0.830 mL) in anhydrous DMF (5 mL) was added dropwise. After stirring for 2 h at 70° C. the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (50 mL) and washed with water (15 mL). The aqueous phase was further extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude product was purified by flash chromatography (eluting with: DCM\MeOH from 95\5 to 90\10) to give the title compound (0.530 g).
m\z ([MH]′)=232.
To a stirred suspension of sodium hydride (0.745 g) in anhydrous DMF (8 mL) cooled at 0° C. a solution of 3-(1H-imidazol-4-yl)-pyridine (3 g) in anhydrous DMF (16 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of methyl 4-chloro-butanoate (2.76 mL) in anhydrous DMF (16 mL) was added dropwise. After stirring for 2.5 h at 70° C. the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (150 mL) and washed with water (50 mL). The aqueous phase was further extracted with EtOAc (3×40 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude product was purified by flash chromatography (eluting with: DCM\MeOH 96\4) to give the title compound (2.2 g).
m\z ([MH]+)=246.
To a stirred suspension of sodium hydride (0.745 g) in anhydrous DMF (8 mL) cooled at 0° C. a solution of 3-(1H-imidazol-4-yl)-pyridine (3 g) in anhydrous DMF (16 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of methyl 5-bromo-pentanoate (3.55 mL) in anhydrous DMF (16 mL) was added dropwise. After stirring for 2 h at 70° C. the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (150 mL) and washed with water (50 mL). The aqueous phase was further extracted with EtOAc (3×40 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude product was purified by flash chromatography (eluting with: DCM\MeOH 96\4) to give the title compound (4.25 g).
m\z ([MH]+)=260.
To a solution of intermediate 43 (0.100 g) in acetone (2 mL) at room temperature 1M NaOH aqueous solution (0.46 mL) was added. The mixture was stirred at room temperature for 20 h then to reflux for 8 h. Solvent evaporation under reduced pressure gave the title compound (0.100 g).
m\z ([MH]+)=204.
To a solution of intermediate 44 (0.100 g) in acetone (1 mL) at room temperature 1M NaOH aqueous solution (0.43 mL) was added. The mixture was stirred at room temperature for 6 h. Solvent evaporation under reduced pressure gave the title compound (0.099 g).
m\z ([MH]+)=218.
To a solution of intermediate 45 (0.100 g) in acetone (1 mL) at room temperature 1 M NaOH aqueous solution (0.40 mL) was added. The mixture was stirred at room temperature for 6 h. Solvent evaporation under reduced pressure gave the title compound (0.096 g).
m\z ([MH]−)=232.
To a solution of intermediate 46 (0.100 g) in acetone (1 mL) at room temperature 1M NaOH aqueous solution (0.38 mL) was added. The mixture was stirred at room temperature for 3 h. Solvent evaporation under reduced pressure gave the title compound (0.095 g).
m\z ([MH]+)=246.
To a stirred suspension of sodium hydride (0.166 g) in anhydrous DMF (3 mL) cooled to 0° C. a solution of 3-(1H-imidazol-4-yl)-pyridine (1 g) in anhydrous DMF (5 mL) was added dropwise under nitrogen atmosphere. The reaction mixture was allowed to reach room temperature and after 15 min a solution of bromoacetaldehyde dimethyl acetal (0.816 mL) in anhydrous DMF (5 mL) was added dropwise. The reaction mixture was stirred at 70° C. for 4 h and at room temperature overnight. After evaporating the solvent under reduced pressure the crude material was purified by flash chromatography (eluting with DCM\MeOH 90\10) to give the title compound (1.15 g).
m\z ([MH]−)=234
TLC: DCM\MeOH 90\10 (Rf=0.5).
To a solution of 3-(1H-imidazol-4-yl)-pyridine (0.350 g) in anhydrous THF (15 mL) cooled at 0° C. acrylaldehyde (0.540 mL) was added dropwise. The resulting solution was stirred at room temperature for 3 days. Solvent evaporation under reduced pressure gave the title compound (0.460 g).
m\z ([MH]+)=202.
To a solution of intermediate 45 (1.2 g) in anhydrous THF (20 mL) cooled at 0° C. lithium aluminum hydride (1M in THF, 2.55 mL) was added dropwise under nitrogen atmosphere. The reaction mixture was stirred for 2 h at room temperature, then water (30 mL) and EtOAc (75 mL) were added. The solvents were evaporated under reduced pressure and the residue extracted with EtOAc (2×75 mL). The combined organic layers were washed with a saturated sodium\potassium tartrate aqueous solution (80 mL), dried over Na2SO4, concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.600 g)
m\z ([MH]+)=218.
To a solution of oxalyl chloride (0.225 mL) in anhydrous DCM (7 mL) cooled to −78° C. under nitrogen atmosphere a solution of DMSO (0.275 mL) in anhydrous DCM (2 mL) was slowly added. After 15 min at −78° C. a solution of intermediate 53 (0.280 g) in anhydrous DCM (5 mL) was dropped within 30 min. The mixture was stirred at 40° C. for 4 h then TEA(0.900 mL) was added. The reaction was allowed to reach room temperature then water (10 mL) was added. The mixture was extracted with DCM (3×20 mL), the organic phase dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.052 g)
m\z ([MH]+)=216.
To a solution of intermediate 46 (3.54 g) in anhydrous THF (40 mL) cooled to 0° C., lithium aluminum hydride (1M in THF, 7.8 mL) was added dropwise under nitrogen atmosphere. The reaction mixture was stirred for 2.5 h at room temperature then water (50 mL), EtOAc (100 mL) and a 28% NH4OH aqueous solution until pH=9 were added. The solvents were evaporated under reduced pressure and the residue dissolved in EtOAc (2×100 mL). The solution was washed with a saturated sodiumpotassium tartrate aqueous solution, dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH from 95\5 to 90\10) to give the title compound (2.32 g).
TLC: DCM\MeOH 95\5 (Rf=0.18)
To a solution of oxalyl chloride (0.755 mL) in anhydrous DCM (25 mL) cooled to −78° C. under nitrogen atmosphere, a solution of DMSO (1.23 mL) in anhydrous DCM (8 mL) was slowly added. After 15 min at −78° C. a solution of intermediate 55 (1 g) in anhydrous DCM (18 mL) was added dropwise over 30 min. The mixture was stirred at −40° C. for 3.5 h then TEA(3.6 mL) was added. The reaction mixture was allowed to reach room temperature then water (50 mL) was added. The mixture was extracted with DCM (3×100 mL), the organic phase dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH 92) to give the title compound (0.635 g)
m\z ([MH]+)=230.
To a solution of (methoxymethyl)triphenylphosphonium chloride (3.27 g) in anhydrous THF (60 mL) cooled to −78° C. sodium bis(trimethylsilyl)amide (1M in THF, 9.5 mL) was added under nitrogen atmosphere and the solution was stirred for 15 min. A solution of 4-quinoline carboxaldehyde (1 g) in anhydrous THF (10 mL) was added and the reaction mixture was stirred at −78° C. for 30 min, at 0° C. for 1.5 h, then at room temperature overnight. The reaction was quenched with water (50 mL) and extracted with EtOAc (75 mL). The aqueous phase was neutralised with a saturated NH4Cl (50ml) aqueous solution and extracted again with EtOAc (2×75 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude material purified by flash chromatography (eluting with: Hexane\EtOAc from 60\40 to 50\50) to give the title compound (0.905 g).
m\z ([MH])=186.
To a solution of [2-(1,3-dioxolan-2-yl)-ethyl]-triphenylphosphonium bromide (10.6 g) in anhydrous THF (150 mL) cooled to −78° C. a solution of sodium bis(trimethylsilyl)amide (1M in THF, 23.9 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred at −78° C. for 30 min then a solution of 4-quinoline carboxaldehyde (2.5 g) in anhydrous THF (30 mL) was added. The mixture was allowed to reach room temperature, DBU (1.8 mL) was added and the mixture stirred for 6 h at 50° C. The reaction was quenched with a saturated NH4Cl aqueous solution of (100 mL) and extracted with EtOAc (3×150 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material filtered on a silica pad (eluting with: cyclohexane\EtOAc 70\30). After evaporating the solvent under reduced pressure, the residue was dissolved in MeOH (20 mL), palladium (10wt. % on carbon powder, 0.360 g) was added and the mixture stirred under hydrogen atmosphere (1.5 atm) for 2 h. The mixture was filtered over a celite pad eluting with MeOH (2×100 mL) and purification by flash chromatography (eluting with: Et2O) gave the title compound (0.700 g).
m\z ([MH]+)=244.
To a solution of intermediate 58 (0.500 g) in acetone (5 mL) a 2N HCl aqueous solution (5 mL) was added and the mixture stirred at 50° C. for 2 h. After evaporation of the solvent under reduced pressure, NH4OH was added to the residue until pH=9. Aqueous solution was extracted with DCM (2×35 mL), the organic phase dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: EtOAc\cyclohexane 80\20) to give the title compound (0.315 g).
m\z ([MH]+)=200.
To a solution of intermediate 59 (0.160 g) in acetone (4 mL) potassium permanganate (0.063 g) was added portionwise within 1 h. After solvent evaporation, water (4 mL) was added and the mixture refluxed for 30 min. The reaction mixture was slowly cooled to 0° C. and kept at 0° C. overnight. The reaction mixture was filtered through a celite pad eluting with DCM (5 mL) and after removing the solvent under vacuum the crude product was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.050 g).
m\z ([MH]+)=216.
To a stirred suspension of [3-(ethoxycarbonyl)-propyl]-triphenylphosphonium bromide (5.6 g) in anhydrous THF (100 mL) cooled to −78° C. a solution of sodium bis(trimethylsilyl)amide (1M in THF, 12 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred at −78° C. for 1 h then a solution of 4-quinoline carboxaldehyde (1.6 g) in anhydrous THF (15 mL) was dropped. The mixture was allowed to reach room temperature, stirred for 2 h then heated to 50° C. and stirred for 4 h. The reaction was quenched with a saturated NH4Cl aqueous solution (50 mL) and extracted with EtOAc (3×100 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by filtration on a silica gel pad (eluting with: cyclohexane\EtOAc 70\30). After evaporating the solvent under reduced pressure, the residue was dissolved in MeOH (10 mL), palladium (10wt. % on carbon powder, 0.060 g) was added and the mixture stirred under hydrogen atmosphere (1.5 atm) for 1 h. The mixture was filtered through a celite pad eluting with MeOH (2×50 mL) and subsequent solvent evaporation under reduced pressure gave the title compound (0.860 g).
m\z ([MH]−)=258.
To a solution of intermediate 61 (0.194 g) in acetone (1 mL) at room temperature 1M NaOH aqueous solution (0.78 mL) was added. The mixture was refluxed for 2 h. Solvent evaporation under reduced pressure gave the title compound (0.181 g).
m\z ([MH]+)=230.
To a solution of intermediate 61 (0.050 g) in anhydrous toluene (1 mL) cooled to −78° C., diisobutylaluminium hydride (1M sol. in toluene, 0.39 mL) was slowly added. The reaction mixture was stirred for 1 h at −78° C., then quenched with 2 mL of a mixture of water (0.25 mL), acetic acid (1 mL) and Et2O (3 mL) at −78° C. The temperature was allowed to reach room temperature then the crude material was filtered through a celite pad eluting with DCM (3×10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.040 g).
m\z ([MH]+)=214.
To a stirred suspension of sodium hydride (0.017 g) in anhydrous DMF (0.250 mL) cooled at 0° C. a solution of 4-phenyl-1H-imidazole (0.100 g) in anhydrous DMF (0.8 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of 3-bromo-propionic acid methyl ester (0.083 mL) in anhydrous DMF (0.3 mL) was added dropwise. After stirring for 2 h at 70° C. the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (5 mL) and washed with water (3 mL). The aqueous phase was further extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give the title compound (0.128 g).
m\z ([MH]−)=231.
To a solution of intermediate 64 (0.034 g) in acetone (0.50 mL) at room temperature a 1M NaOH aqueous solution (0.15 mL) was added. The mixture was stirred at room temperature for 3 h. Solvent evaporation under reduced pressure gave the title compound (0.035 g).
m\z ([MH]−)=217.
To a stirred suspension of sodium hydride (0.037 g) in anhydrous DMF (0.550 mL) cooled at 0° C. a solution of 4-phenyl-1H-imidazole (0.200 g) in anhydrous DMF (1.6 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of 2-chloro-acetic acid methyl ester (0.134 mL) in anhydrous DMF (0.60 mL) was added dropwise. After stirring for 2 h at 70° C. and overnight at room temperature the mixture was diluted with Et2O (25 mL) and washed with water (10 mL). The aqueous phase was further extracted with Et2O (3×15 mL). The collected organic extracts were dried over Na2SO4, concentrated under reduced pressure and the crude product was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 96\4) to give the title compound (0.210 g).
m\z ([MH]−)=217.
To a solution of intermediate 66 (0.190 g) in acetone (2.5 mL) at room temperature, a 1.2M NaOH aqueous solution (0.730 mL) was added. The mixture was stirred at room temperature for 2.5 h and then solvent evaporation under reduced pressure gave the title compound (0.205 g).
m\z ([MH]−)=203
To a stirred suspension of sodium hydride (0.013 g) in anhydrous DMF (0.250 mL) cooled at 0° C. a solution of 4-thiophen-2-yl-1H-imidazole (0.100 g) in anhydrous DMF (1.6 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of methyl 3-bromopropionate (0.100 mL) in anhydrous DMF (0.3 mL) was added dropwise. After stirring for 5 h at 70° C. and overnight at room temperature the solvent was evaporated under reduced pressure, the residue dissolved with EtOAc (5 mL) and the solution washed with water (3 mL). The aqueous phase was further extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure and the crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give the title compound (0.013 g).
m\z ([MH]−)=237.
To a solution of intermediate 68 (0.013 g) in acetone (0.3 mL) at room temperature, a 1M NaOH aqueous solution (0.055 mL) was added. The mixture was stirred at room temperature for 5 h and then solvent evaporation under reduced pressure gave the title compound (0.012 g).
m\z ([MH]−)=223.
To a stirred suspension of sodium hydride (0.016 g) in anhydrous DMF (0.3 mL) cooled at 0° C. a solution of 2-(1H-pyrazol-3-yl)-thiazole (0.100 g) in anhydrous DMF (0.8 mL) was added dropwise under nitrogen atmosphere. The mixture was stirred for 15 min at room temperature then a solution of 3-bromo-propionic acid methyl ester (0.080 mL) in anhydrous DMF (0.3 mL) was added dropwise. After stirring for 4 h at 70° C. the solvent was evaporated under reduced pressure, the residue diluted with EtOAc (5 mL) and washed with water (3 mL). The aqueous phase was further extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude material was dissolved in acetone (1 mL) and a 1.2N NaOH aqueous solution (0.55 mL) was added. The mixture was stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.025 g).
m\z ([MH]−)=224.
To a solution of 2-chloro-3-methoxy-quinoxaline (0.100 g) in anhydrous DMF (3 mL) potassium carbonate (0.142 g) and mercapto-acetic acid ethyl ester (0.084 mL) were added and the resulting mixture was stirred at 80° C. for 1.5 h. The solvent was removed under reduced pressure, water (5 mL) was added and the mixture was extracted with EtOAc (3×10 mL). The combined organic phases were washed with brine (5 mL), dried over Na2SO4 and evaporated under reduced pressure. The crude material was purified by flash chromatography (eluting with: cyclohexane\EtOAc 80\20) to afford the title compound (0.065 g).
m\z ([MH]−)=255
TLC: cyclohexane\EtOAc 80\20 (Rf=0.57).
To a solution of intermediate 71 (0.060 g) in THF (2 mL) a 3N NaOH aqueous solution (2 mL) was added. The mixture was vigorously stirred at room temperature overnight, then after solvent evaporation, a 1N HCl aqueous solution was added until pH=1. The solution was extracted with DCM (3×8 mL), the organic phase was washed with brine (5 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.045 g).
m\z ([MH]+)=227
TLC: EtOAc\MeOH 98\2 (Rf=0.48)
To a solution of quinoxalin-2-ol (2 g) in acetone (30 mL) potassium carbonate (3.8 g) and chloroacetic acid ethyl ester (2.2 mL) were added and the resulting mixture was stirred at reflux for 6 h. After evaporating the solvent under reduced pressure, water (30 mL) was added and the mixture extracted with EtOAc (3×50 mL). The organic phase was washed with brine (30 mL) dried over Na2SO4 and evaporated under reduced pressure. The crude material was purified by flash chromatography (eluting with: cyclohexane\EtOAc 3\2) to afford the title compound (1.5 g).
m\z ([MH]+)=233.
To a solution of intermediate 73 (0.300 g) in THF (10 mL) a 3N NaOH aqueous solution (10 mL) was added. The mixture was vigorously stirred at room temperature for 1 h, then after evaporation of the solvent, a 1N HCl aqueous solution was added until pH=1. The aqueous solution was extracted with DCM (3×20 mL), the organic phase was washed with brine (10 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.230 g).
m\z ([MH]+)=205
To a solution of 4-(3,5-difluoro-phenyl)-1H-pyrazole (0.050 g) in anhydrous acetonitrile (2 mL), acrylaldehyde (0.050 mL) was added portionwise. The resulting solution was stirred at 55° C. for 16 h and then the solvent was removed under reduced pressure to give the title compound (0.52 g).
m\z ([MH]+)=237
To a solution of 4-(4-chloro-phenyl)-2,5-dimethyl-1H-imidazole (0.030 g) in anhydrous acetonitrile (3 mL), acrylaldehyde (0.069 mL) was added portionwise. The resulting solution was stirred at 50° C. for 24 h and then the solvent was removed under reduced pressure to give the title compound (0.036 g).
m\z ([MH]+)=263
To a solution of 4-(4-nitro-phenyl)1H-imidazole (0.100 g) in anhydrous acetonitrile (12 mL), acrylaldehyde (0.450 mL) was added portionwise. The resulting solution was stirred at 50° C. for 5 days. Solvent was removed under reduced pressure to give the title compound (0.120 g).
m\z ([MH]+)=246
To a solution of 4-(1H-imidazol-4-yl)-pyridine (0.040 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.050 mL) was added portionwise. The resulting solution was stirred at 55° C. for 8 h and at room temperature overnight. Solvent was removed under reduced pressure to give the title compound (0.42 g).
m\z ([MH]+)=202
To a solution of oxalyl chloride (0.127 mL) in anhydrous DCM (4 mL) cooled to −78° C. under nitrogen atmosphere DMSO (0.155 mL) was slowly added. After stirring for 30 min at −78° C. a solution of 3-(3-trifluoromethyl-1H-pyrazol-4-yl)-propan-1-ol (0.094 mL) in anhydrous DCM (3 mL) was added dropwise. The mixture was stirred at −40° C. for 3 h then TEA(0.507 mL) was added. The reaction was allowed to reach room temperature then a saturated NaHCO3 aqueous solution (10 mL) was added and the mixture extracted with DCM (3×10 mL). The organic phase was washed with brine (10 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.100 g)
m\z ([MH]′)=193
To a solution of 5-methyl-4-(4-trifluoromethyl-phenyl)-1H-imidazole (0.030 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.027 mL) was added. The resulting solution was stirred at 50° C. for 24 h and at room temperature for 12 h. Solvent was removed under reduced pressure to give the title compound (0.038 g).
m\z ([MH]+)=283
To a solution of oxalyl chloride (0.127 mL) in anhydrous DCM (4 mL) cooled to −78° C. under nitrogen atmosphere DMSO (0.155 mL) was slowly added. After 20 min at −78° C. a solution 3-pyridin-3-yl-propan-1-ol (0.100 g) in anhydrous DCM (3 mL) was added dropwise within 30 min. The mixture was stirred at 40° C. for 3 h then TEA(0.507 mL) was added. The reaction was allowed to reach room temperature then water (5 mL) was added. The mixture was extracted with DCM (3×10 mL), the organic phase dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give the title compound (0.052 g)
m\z ([MH]+)=136
To a solution of 2-(1H-pyrazol-4-yl)-pyridine (0.058 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.022 mL) was added. The resulting solution was stirred at 50° C. for 3 h. Solvent was removed under reduced pressure to give the title compound (0.060 g).
m\z ([MH]+)=202
To a solution of oxalyl chloride (0.127 mL) in anhydrous DCM (3 mL) cooled to −70° C. under nitrogen atmosphere, a solution of DMSO (0.155 mL) in anhydrous DCM (2 mL) was added dropwise. The reaction mixture was stirred for 30 min then a solution of 3-pyridin-4-yl-propan-1-ol (0.100 g) in anhydrous DCM (2 mL) was added dropwise in 30 min. The mixture was stirred at −60° C. for 3 h then the reaction mixture was allowed to reach −10° C. and TEA (0.507 mL) was added. The reaction was stirred overnight reaching slowly the room temperature. A saturated NaHCO3 aqueous solution (5 mL) was added, the organic phase was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.100 g).
m\z ([MH]+)=136
To a solution of 4-(1H-pyrazol-4-yl)-pyrimidine (0.022 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.020 mL) was added. The reaction mixture was stirred at 50° C. for 1.5 h. Solvent was removed under reduced pressure to give the title compound (0.025 g).
m\z ([MH]+)=203
To a solution of 2-(1H-pyrazol-4-yl)-pyridine (0.050 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.220 mL) was added. The resulting solution was stirred at 50° C. for 4 h and at room temperature overnight. Solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=202
To a solution of 4-(3,5-dichloro-phenyl)-2,5-dimethyl-1H-imidazole (0.035 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.116 mL) was added portionwise. The resulting solution was stirred at 50° C. for 56 h. Solvent was removed under reduced pressure to give the title compound (0.029 g).
m\z ([MH]+)=297
To a solution of 2,5-dimethyl-4-(3-trifluoromethyl-phenyl)-1H-imidazole (0.037 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.120 mL) was added portionwise. The resulting solution was stirred at 50° C. for 48 h. Solvent was removed under reduced pressure to give the title compound (0.042 g).
m\z ([MH]+)=297
To a stirred suspension of 2-(1H-pyrazol-4-yl)-1,3-benzoxazole (0.053 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.220 mL) was added. The reaction mixture was stirred at 50° C. for 6 h. After filtration the solvent was removed under reduced pressure to give the title compound (0.034 g).
m\z ([MH]+)=242
To a solution of 4-(1H-pyrazol-3-yl)-pyridine (0.030 g) in anhydrous acetonitrile (1 mL) acrylaldehyde (0.085 mL) was added. The reaction mixture was stirred at 50° C. for 48 h. Solvent was removed under reduced pressure to give the title compound (0.036 g).
m\z ([MH]+)=202.
To a solution of 4-(1H-imidazol-2-yl)-pyridine (0.050 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.080 mL) was added. The reaction mixture was stirred at 50° C. for 14 h. Solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=202
To a solution of 2-(1H-pyrazol-4-yl)quinoline (0.050 g) in anhydrous acetonitrile (5 mL) acrylaldehyde (0.116 mL) was added. The reaction mixture was stirred at 50° C. for 3 days. Solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=252
To a solution of 4-(1H-pyrazol-4-yl)quinoline (0.050 g) in anhydrous acetonitrile (4 mL) acrylaldehyde (0.260 mL) was added. The reaction mixture was stirred at 50° C. for 7 days. Solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=252
To a solution of 2-(1H-pyrazol-4-yl)quinoxaline (0.050 g) in anhydrous acetonitrile (4 mL) acrylaldehyde (0.156 mL) was added. The reaction mixture was stirred at 50° C. for 3 days. Solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=253
To a solution of oxalyl chloride (0.190 mL) in anhydrous DCM (6 mL) cooled to −78° C. under nitrogen atmosphere a solution of DMSO (0.230 mL) in anhydrous DCM (4 mL) was slowly added. After stirring for 30 min at −78° C. a solution of 3-thien-3-ylpropan-1-ol (0.150 g) in anhydrous DCM (4 mL) was added dropwise. The mixture was stirred at −40° C. for 3 h then TEA(0.750 mL) was added. The reaction was allowed to reach room temperature then a saturated NaHCO3 aqueous solution (10 mL) was added and the mixture extracted with DCM (3×10 mL). The organic phase was washed with brine (10 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.140 g)
m\z ([MH]+)=141
To a solution of 2-methyl-3-(2H-pyrazol-5-yl)-pyrazine (0.030 g) in anhydrous acetonitrile (1 mL) acrylaldehyde (0.065 mL) was added. The reaction mixture was stirred at 50° C. for 60 h. Solvent was removed under reduced pressure to give the title compound (0.038 g).
m\z ([MH]+)=217.
To a solution of 2-(methylthio)-1H-benzimidazole (0.030 g) in anhydrous acetonitrile (1 mL) acrylaldehyde (0.080 mL) was added. The reaction mixture was stirred at 50° C. for 48 h. Solvent was removed under reduced pressure to give the title compound (0.039 g).
m\z ([MH]+)=221
To a stirred suspension of Dess-Martin periodinane (0.163 g) in anhydrous DCM (4 mL) 3-[3-(4-chlorophenyl)-1H-pyrazol-5-yl]propan-1-ol (0.050 g) was added and the mixture was stirred at room temperature overnight. The reaction was quenched with a Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 3 mL), stirred for lh then extracted with DCM (10 mL). The organic phase washed with brine (5 mL), separated, dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.040 g).
m\z ([MH]+)=235
To a solution of 6-(methylthio)-7H-purine (0.032 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.040 mL) was added. The reaction mixture was stirred at 80° C. for 8 h and overnight at room temperature. Solvent was removed under reduced pressure to give the title compound (0.042 g).
m\z ([MH]+)=223
To a solution of 6-methoxy-7H-purine (0.040 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.057 mL) was added. The reaction mixture was stirred at 80° C. for 8 h. Solvent was removed under reduced pressure to give the title compound (0.055 g).
m\z ([MH]+)=207
To a solution of 6-methoxy-1,3-benzoxazol-2(3H)-one (0.072 g) in anhydrous acetonitrile (2 mL) was added acrylaldehyde (0.060 mL). The reaction mixture was stirred at 50° C. for 3 h. Solvent was removed under reduced pressure to give the title compound (0.080 g).
m\z ([MH]+)=222
To a solution of 1H-pyrrolo[2,3-b]pyridine (0.040 g) in anhydrous acetonitrile (3 mL), acrylaldehyde (0.071 mL) was added. The reaction mixture was stirred at 80° C. for 6 h and then the solvent was removed under reduced pressure to give the title compound (0.042 g).
m\z ([MH]+)=175
To a solution of 2,4-dimethyl-5-(1H-pyrazol-3-yl)-1,3-thiazole (0.032 g) in anhydrous acetonitrile (1 mL) acrylaldehyde (0.040 mL) was added. The reaction mixture was stirred at 50° C. for 5 h and then at room temperature for 3 days. Solvent evaporation under reduced pressure gave the title compound (0.042 g).
m\z ([MH]+)=236
To a solution of 3H-imidazo[4,5-c]pyridine (0.050 g) in anhydrous acetonitrile (4 mL) acrylaldehyde (0.025 mL) was added. The reaction mixture was stirred at 80° C. overnight and then solvent was removed under reduced pressure to give a 1\1 mixture of the title compounds (0.050 g).
m\z ([MH]+)=176
To a solution of 1H-benzimidazole (0.050 g) in anhydrous acetonitrile (4 mL) acrylaldehyde (0.160 mL) was added. The reaction mixture was stirred at 80° C. overnight and then the solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=175
To a solution of 3H-imidazo[4,5-b]pyridine (0.050 g) in anhydrous acetonitrile (3 mL) acrylaldehyde (0.250 mL) was added. The reaction mixture was stirred at 80° C. for 24 h and then the solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=176
To a solution of 2-quinoxalinethiol (0.200 g) in a mixture of anhydrous dioxane\DMF 4\1 (5 mL) sodium hydride (80% mineral oil, 0.044 g) was added portionwise under a nitrogen atmosphere and the reaction mixture stirred at room temperature for 15 min. Then 3-bromopropionaldheyde dimethyl acetal (0.200 mL) was added and the solution was heated to 80° C. for 1.5 h. After dilution at room temperature with EtOAc (5 mL), the solution was concentrated under reduced pressure, diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic phases were then washed with brine (10 mL), dried over Na2SO4, and concentrated under vacuum to give a crude material that was purified by flash chromatography (eluant: cyclohexane\EtOAc 3\1) to afford the title compound (0.280 g).
m\z ([MH]+)=265.
TLC: Cyclohexane\EtOAc 7\3 (Rf=0.47).
To a solution of 4-phenyl-1H-imidazole (0.050 g) in anhydrous acetonitrile (4 mL) acrylaldehyde (0.210 mL) was added. The reaction mixture was stirred at 80° C. for 24 h and then the solvent was removed under reduced pressure to give the title compound (0.050 g).
m\z ([MH]+)=201.
To a stirred suspension of sodium hydride (0.009 g) in anhydrous DMF (0.5 mL) 4-(1H-imidazol-4-yl)pyridine (0.040 g) was added under nitrogen atmosphere and the reaction mixture was stirred at room temperature for 30 min. Then 2-bromoacetaldheyde dimethyl acetal (0.040 mL) was added and the solution was heated to 70° C. for 8 h. After cooling to room temperature the solvent was evaporated to give the title compound (0.064 g)
m\z ([MH]+)=234
To a solution of 2-quinoxalinethiol (0.200 g) in a mixture of anhydrous dioxane\DMF 4\1 (5 mL) sodium hydride (80% mineral oil, 0.044 g) was added portionwise under nitrogen atmosphere and the reaction mixture stirred at room temperature for 15 min. Then 2-bromoacetaldheyde dimethyl acetal (0.175 mL) was added and the solution was heated to 80° C. for 4.5 h. After dilution at room temperature with EtOAc (5 mL), the solution was concentrated under reduced pressure, diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic phases were washed with brine (10 mL), dried over Na2SO4, evaporated under vacuum obtaining a crude product that was purified by flash chromatography (eluting with: Cyclohexane\EtOAc 3\1) afforded the title compound (0.167 g).
m\z ([MH]+)=251.
TLC: Cyclohexane\EtOAc 7\3 (Rf=0.52).
To a solution of 4-thiophen-2-yl-1H-imidazole (0.030 g) in anhydrous acetonitrile (3 mL) TEA(0.022 mL) was added. After stirring at room temperature for 15 min acrylaldehyde (0.034 mL) was added dropwise and the resulting solution was heated to 75° C. for 8 h and then the solvent was removed under reduced pressure to give the title compound (0.033 g).
m\z ([MH]+)=207.
To a solution of 6-methyl-1,3-benzoxazol-2(3H)-one (0.030 g) in anhydrous acetonitrile (2 mL) acrylaldehyde (0.060 mL) was added. The reaction mixture was stirred at 50° C. for 48 h and then the solvent was removed under reduced pressure to give the title compound (0.040 g).
m\z ([MH]+)=206.
To a stirred suspension of sodium hydride (0.009 g) in anhydrous DMF (0.5 mL) 4-(1H-imidazol-2-yl)pyridine (0.040 g) was added under nitrogen atmosphere and the reaction mixture was stirred at room temperature for 30 min. Then 2-bromoacetaldheyde dimethyl acetal (0.040 mL) was added and the solution was heated to 70° C. for 30 h. After cooling to room temperature the solvent was evaporated under vacuum to give the title compound (0.064 g).
m\z ([MH]+)=234
To a solution of 3,4-dimethyl-5-(1H-1,2,4-triazol-3-yl)thiophene-2-carbonitrile (0.037 g) in anhydrous acetonitrile (1 mL) acrylaldehyde (0.040 mL) was added and the resulting solution was heated to 50° C. for 5 h. Solvent evaporation under vacuum gave the title compound (0.046 g).
m\z ([MH]+)=261
To a solution of oxalyl chloride (0.290 mL) in anhydrous DCM (8 mL) cooled to −78° C. under nitrogen atmosphere DMSO (0.340 mL) was slowly added. After 1 h at −78° C. a solution of 3-quinolin-3-ylpropan-1-ol (0.300 g) in anhydrous DCM (3 mL) was added. After stirring at −40° C. for 3 h TEA(0.892 mL) was added and the reaction mixture was allowed to reach room temperature. Water (10 mL) was added and the mixture was extracted with DCM (3×10 mL), the organic phase dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.290 g)
m\z ([MH]+)=186.
To a solution of 4-(3-nitrophenyl)-1H-imidazole (0.040 g) in anhydrous acetonitrile (5 mL) acrylaldehyde (0.120 mL) was added and the resulting solution was heated to 80° C. and stirred for 4 days. The solvent was removed under reduced pressure to give the title compound (0.041 g).
m\z ([MH]+)=246.
To a solution of lithium aluminum hydride (1M in THF, 9.53 mL) in anhydrous THF (15 mL) cooled to 0° C. a solution of 4-(2,3-dihydro-1,4-benzodioxin-6-yl)-4-oxobutyric acid (0.750 g) in anhydrous THF (5 mL) was added dropwise. The reaction mixture was heated to reflux for 24 h, then it was cooled to room temperature and diluted with EtOAc (10 mL). After evaporation of the solvents under reduced pressure the crude material was treated with a 1N HCl aqueous solution (20 mL) and DCM (100 mL). The organic phase was washed with a saturated NaHCO3 aqueous solution (50 mL), brine (50 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (EtOAc\cyclohexane from 50\50 to 100\0) to give the title compound (0.340 g).
m\z ([MH]−)=225
To a solution of oxalyl chloride (0.560 mL) in anhydrous DCM (5 mL) cooled to −78° C. under nitrogen atmosphere a solution of DMSO (0.910 mL) in anhydrous DCM (5 mL) was slowly added. After stirring for 30 min at −78° C. a solution of intermediate 116 (0.340 g) in anhydrous DCM (2 mL) was added dropwise. The reaction mixture was stirred at 40° C. for 3 h then TEA(2.5 mL) was added and then the reaction mixture was allowed to reach room temperature. A saturated NaHCO3 aqueous solution (20 mL) was added and the mixture extracted with DCM (3×20 mL). The organic phase was washed with brine (20 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (EtOAc\cyclohexane 40\60) to give the title compound (0.250 g).
m\z ([MH]+)=221
To a solution of 5-methoxy-1H-pyrrolo[3,2-b]pyridine (0.040 g) in anhydrous THF (1.5 mL) cooled to 0° C., sodium hydride (0.010 g) was added under nitrogen atmosphere. The mixture was stirred at room temperature for 1 h then 2-(2-bromoethyl)-1,3-dioxolane (0.038 mL) was added and the stirring continued for an additional 6 h. The reaction mixture was quenched with water (3 mL) and extracted with DCM (3×5 mL), the organic phase dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.067).
m\z ([MH]+)=249.
To a stirred suspension of sodium hydride (0.005 g) in anhydrous DMF (0.5 mL) at 0° C., under nitrogen atmosphere, a solution of 2-(1H-pyrazol-3-yl)-1,3-thiazole (0.030 g) in anhydrous DMF (1 mL) was added. The mixture was allowed to reach room temperature and stirred for an additional 15 min and then a solution of bromoacetaldehyde dimethylacetal (0.024 mL) in anhydrous DMF (0.5 mL) was added. After stirring the reaction mixture at room temperature for 24 h, the solvent was evaporated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM) to give the title compound (0.015 g).
m\z ([MH]+)=240.
To a stirred suspension of sodium hydride (0.005 g) in anhydrous DMF (0.7 mL) at 0° C. a solution of 4-phenyl-1H-imidazole (0.023 g) in anhydrous DMF (1 mL) was added under nitrogen atmosphere. The mixture was allowed to reach room temperature and stirred for an additional 15 min, then a solution of bromoacetaldehyde dimethylacetal (0.022 mL) in anhydrous DMF (0.7 mL) was added. After stirring the reaction mixture at room temperature for 24 h the solvent was evaporated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 90\10) to give the title compound (0.027 g).
m\z ([MH]+)=233.
To a stirred suspension of sodium hydride (0.019 g) in anhydrous DMF (3 mL) at 0° C., under nitrogen atmosphere, a solution of 4-thien-2-yl-1H-imidazole (0.100 g) in anhydrous DMF (1 mL) was added. The mixture was allowed to reach room temperature, stirred for an additional 15 min and then a solution of bromoacetaldehyde dimethylacetal (0.090 mL) in anhydrous DMF (1 mL) was added. After stirring at room temperature for 24 h the solvent was evaporated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 99\1) to give the title compound (0.088 g).
m\z ([MH]+)=239.
To a solution of intermediate 3 (0.060 g) in anhydrous DMF (16 mL) potassium cyanide (0.051 g) was added under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h, quenched with a saturated NaHCO3 aqueous solution (30 mL) and extracted with DCM (3×30 mL). The organic phase was then washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.017 g).
1H-NMR (CDCl3) δ: 5.27(dd, 1H), 4.74 (dd, 1H), 4.62 (d, 1H), 4.42 (d, 1H), 4.26 (d, 1H), 3.84 (q, 1H), 3.56 (m, 1H), 3.16 (m, 1H), 3.10-3.0 (m, 2H), 2.77(s, 3H), 2.68 (m, 1H), 2.60 (m, 1H), 2.25 (s, 6H), 2.05 (m, 3H), 1.90 (m, 1H), 1.68 (m, 1H), 1.63 (m, 2H), 1.56 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.30 (s, 3H), 1.26 (d, 3H), 1.18 (d, 3H), 1.14 (d, 3H), 1.06 (d, 3H), 0.92 (t, 3H).
TLC: DCM\MeOH 95\5 (Rf=0.57).
A solution of example 1 (0.024 g) in MeOH (1 mL) was stirred for 24 h then concentrated under reduced pressure to give the title compound (0.020 g).
1H-NMR (CDCl3) δ: 5.26 (dd, 1H), 4.61 (d, 1H), 4.34 (d, 1H), 4.28 (m, 1H), 3.87 (q, 1H), 3.57 (m, 1H), 3.18 (m, 1H), 3.15 (t, 1H), 3.12 (m, 1H), 3.06 (m, 1H), 2.78 (s, 3H), 2.62 (m, 1H), 2.46 (m, 1H), 2.27 (s, 6H), 1.91 (m, 1H), 1.84 (m, 1H), 1.70 (m, 1H), 1.68 (m, 1H), 1.62 (m, 1H), 1.57 (s, 3H), 1.41 (d, 3H), 1.34 (d, 3H), 1.34 (s, 3H), 1.24 (m, 1H), 1.26 (d, 3H), 1.14 (d, 3H), 1.07 (d, 3H), 0.92 (t, 3H).
TLC: DCM\MeOH 90\10 (Rf=0.38).
To a solution of intermediate 4 (0.036 g) in iPrOH (1.5 mL), sodium cyanoborohydride (0.023 g) and titanium(III) chloride (10 wt. % solution in 20\30 wt. % hydrochloric acid, 0.1 mL) were added portionwise within 6 h. The mixture was diluted with a saturated NaHCO3 aqueous solution (3×2 mL), extracted with DCM (3×2 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH 100\0\0, 98\2\0, 95\5\0.5) to give the title compound (0.010 g).
1H-NMR (CDCl3) δ: 5.06 (dd, 1H), 4.76 (dd, 1H), 4.38 (d, 1H), 4.17 (d, 1H), 3.80 (q, 1H), 3.54 (bm, 1H), 3.24 (m, 1H), 3.20-3.00 (m, 4H), 2.64 (s, 3H), 2.60 (m, 1H), 2.54 (m, 1H), 2.25 (s, 1H+6H), 2.08 (s, 3H), 1.94 (m, 1H), 1.74 (m, 1H), 1.70-1.50 (m, 3H), 1.40 (d, 3H), 1.30 (m, 1H), 1.30(s, 3H), 1.26 (d, 3H), 1.17 (d, 3H), 1.16 (d, 3H), 1.12 (d, 3H), 0.86 (t, 3H).
To a solution of intermediate 18 (1.5 g) and EDC (3.1 g) in DCM (100 mL) cooled to 0° C., DMSO (3.45 mL) was added under nitrogen atmosphere. After stirring at 0° C. for 10 min, a solution of pyridinium trifluoroacetate (3.12 g) in anhydrous DCM (15 mL) was slowly added. After 10 min the ice-bath was removed. The reaction mixture was stirred for 3 h at room temperature then quenched with water (150 mL) and extracted with DCM (3×250 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (1.2 g).
1H-NMR (CDCl3) δ: 7.80-7.20 (m, 10H), 6.40 (dd, 1H), 5.15 (s, 1H), 4.73 (m, 1H), 4.42 (d, 1H), 4.16 (d, 1H), 3.90 (q, 1H), 3.55 (m, 1H), 3.17 (m, 1H), 2.95 (m, 1H), 2.94 (d, 1H), 2.67 (m, 1H), 2.53 (s, 3H), 2.43 (m, 1H), 2.33 (s, 6H), 2.05 (s, 3H), 2.00 (m, 1H), 1.74 (m, 1H), 1.65 (m, 1H), 1.53 (s, 3H), 1.38 (d, 3H), 1.23 (s, 3H), 1.29 (d, 3H), 1.25 (m, 1H), 1.25 (d, 3H), 1.14 (d, 3H), 1.07 (d, 3H), 0.83 (t, 3H).
TLC: DCM\MeOH 10\1 (Rf=0.30).
A solution of example 4 (0.030 g) in MeOH (1 mL) was stirred at room temperature overnight.
Solvent evaporation under reduced pressure gave the title compound (0.024 g).
1H-NMR (CDCl3) δ: 7.80-7.20 (m, 10H), 6.38 (dd, 1H), 5.13 (s, 1H), 4.34 (d, 1H), 4.18 (d, 1H), 3.91 (q, 1H), 3.55 (m, 1H), 3.20 (m, 1H), 2.96 (m, 1H), 2.63 (s, 1H), 2.53 (s, 1H), 2.44 (m, 2H), 2.26 (s, 6H), 2.00 (m, 1H), 1.70-1.45 (m, 7H), 1.45 (d, 1H), 1.30 (m, 4H), 1.04 (d, 3H), 0.96 (t, 3H), 0.91 (d, 3H).
TLC: DCM\MeOH 10\1 (Rf=0.34).
A solution of example 4 (1.1 g) in acetonitrile (30 mL) and a 1.2N HCl aqueous solution (70 mL) was stirred at room temperature for 1 h. After neutralising the mixture with solid Na2CO3 and evaporating the solvent, the aqueous phase was extracted with DCM (3×200 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.9 g).
1H-NMR (CDCl3) δ: 5.45 (dd, 1H), 4.75 (m, 1H), 4.45 (d, 1H), 4.40 (d, 1H), 4.21 (d, 1H), 3.82 (q, 1H), 3.54 (m, 1H), 3.09 (m, 1H), 2.69 (m, 1H), 2.68 (s, 3H), 2.58 (m, 1H), 2.41(m, 1H), 2.25 (s, 6H), 2.07 (m, 3H), 1.95 (m, 1H), 1.75 (m, 1H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.31 (s, 3H), 1.26 (d, 3H), 1.17 (d+d, 6H), 1.09 (d, 3H), 0.88 (t, 3H).
TLC: DCM\MeOH 10\1 (Rf=0.48).
To a solution of example 6 (15.1 g) in anhydrous toluene (170 mL) fresh distilled benzaldehyde (2.52 mL) and pyridine (2.01 mL) were added and the reaction mixture was refluxed for 4.5 h. After evaporating the solvent the residue was dissolved in acetonitrile (60 mL) then a 1.2N HCl aqueous solution (120 mL) was added at room temperature. After stirring for 1.5 h the solvent was evaporated and the aqueous acid solution extracted with EtOAc (150 mL) and DCM (150 mL). The aqueous phase was neutralized with solid potassium carbonate and extracted with EtOAc (2×50 mL). The collected organic extracts were dried over Na2SO4 and concentrated under reduced pressure to give the title compound (11.5 g, 96% pure by NMR analysis).
1H-NMR (CDCl3) δ: 5.45 (dd, 1H), 4.75 (m, 1H), 4.45 (d, 1H), 4.40 (d, 1H), 4.21 (d, 1H), 3.82 (q, 1H), 3.54 (m, 1H), 3.09 (m, 1H), 2.69 (m, 1H), 2.68 (s, 3H), 2.58 (m, 1H), 2.41(m, 1H), 2.25 (s, 6H), 2.07 (m, 3H), 1.95 (m, 1H), 1.75 (m, 1H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.31 (s, 3H), 1.26 (d, 3H), 1.17 (d+d, 6H), 1.09 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.012 g) in MeOH (1 mL) was stirred at room temperature overnight. After evaporating the solvent under reduced pressure the crude material was purified by flash chromatography (eluting with: DCM\MeOH 100\5) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 5.41 (dd, 1H), 4.42 (d, 1H), 4.33 (d, 1H), 4.22 (d, 1H), 3.83 (q, 1H), 3.56 (m, 1H), 3.23 (d, 1H), 3.12 (m, 1H), 3.02 (m, 1H), 2.80 (d, 1H), 2.69 (s, 3H), 2.60 (m, 1H), 2.54 (m, 1H), 2.40 (d, 1H), 2.33 (s, 6H), 1.95 (m, 1H), 1.9-1.50 (m, 3H), 1.49 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.33 (s, 3H), 1.32 (m, 1H), 1.31 (d, 3H), 1.26 (d, 3H), 1.16(d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
To a solution of intermediate 23 (0.050 g) in anhydrous DCM (25 mL), EDC (0.102 g) and DMSO (0.115 mL) were added under nitrogen atmosphere. The mixture was cooled to 0° C. and a solution of pyridinium trifluoroacetate (0.102 g) in DCM (0.5 mL) was added dropwise. The mixture was allowed to reach room temperature; after stirring for 5 h water (10 mL) was added and the mixture extracted with DCM (2×20 mL). The organic phase was dried over Na2SO4, concentrated under reduced pressure and the crude material purified by preparative TLC (eluting with: DCM\MeOH 95\5). The recovered silica gel was stirred for 18 h in MeOH (2 mL), the mixture was filtered and solvent evaporation under reduced pressure gave the title compound (0.025 g).
1H-NMR (CDCl3) δ: 4.90 (dd, 1H), 4.32 (d, 1H), 4.24 (d, 1H), 3.85 (q, 1H), 3.56 (m, 1H), 3.32 (d, 1H), 3.18 (m, 1H), 3,12 (m, 1H), 3.02 (m, 1H), 2.80 (d, 1H), 2.71 (dd, 1H), 2.63 (s, 3H), 2.55 (m, 1H), 2.47 (m, 1H), 2.27 (s, 6H), 1.87 (m, 1H), 1.70 (m, 1H), 1.62 (m, 1H), 1.58 (m, 1H), 1.50 (s, 3H), 1.38 (d, 3H), 1.30 (d, 3H), 1.31 (s, 3H), 1.30 (m, 1H), 1.25 (d, 3H), 1.22 (m, 1H), 1.14 (d, 3H), 1.07 (d, 3H), 0.86 (t, 3H).
To a solution of example 7 (0.284 g) in anhydrous THF (14 mL) cooled to −10° C. potassium tert-butoxide (1M in THF, 0.553 mL) was added under nitrogen atmosphere. After 5 min N-fluorobenzenesulfonimide (0.148 g) was added at −10° C. The mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM (15 mL) and washed with water (15 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0, to 94\6\0.2) to give the title compound (0.120 g).
m\z ([MH]+)=687.
1H-NMR (CDCl3) δ: 5.35 (dd, 1H), 4.74 (M, 1H), 4.48 (d, 1H), 4.33 (d, 1H), 3.96 (m, 1H), 3.52 (m, 1H), 2.89 (s, 3H), 2.86 (m, 1H), 2.72 (m, 1H), 2.53 (m, 1H), 2.27 (s, 6H), 2.08 (s, 3H), 2.0-1.94 (m, 2H), 1.82 (d, 3H), 1.76 (m, 1H), 1.64 (m, 1H), 1.48 (d+m, 3H+1H), 1.31 (s, 3H), 1.26 (m, 6H), 1.16 (d, 3H), 1.08 (d, 3H), 0.92 (t, 3H).
A solution of example 10 (0.010 g) in MeOH (1 mL) was stirred at room temperature overnight. Evaporation of the solvent under reduced pressure gave the title compound (0.005 g).
1H-NMR (CDCl3) δ: 5.33 (dd, 1H), 4.42 (d, 1H), 4.33 (d, 1H), 4.03 (dd, 1H), 3.53 (bm, 1H), 3.18 (m, 1H), 3.10 (m, 1H), 2.93 (s, 3H), 2.90 (m, 1H), 2.54 (m, 2H), 2.30 (s, 6H), 2.09 (s, 1H), 2.06 (m, 1H), 1.96 (m, 1H), 1.82 (d, 1H), 1.62-1.50 (m, 6H), 1.34 (s, 3H), 1.27(m, 4H), 1.26 (s, 3H), 1.16 (d, 3H), 1.08 (d, 3H), 0.92 (t, 3H).
To a solution of example 7 (1.34 g) in anhydrous acetonitrile (12 mL) tert-butyl N-(2-oxoethyl)carbamate (0.640 g) was added under nitrogen atmosphere and the resulting mixture was stirred for 24 h at room temperature. Sodium cyanoborohydride (1M in THF, 2.0 mL) and acetic acid (0.114 mL) were added and the reaction mixture was stirred for 12 h at room temperature. After evaporating the solvent, the residue was dissolved in DCM (100 mL) and washed with a saturated NaHCO3 aqueous solution (50 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The residue was diluted in MeOH (35 mL) and refluxed for 15 h. The solvent was evaporated under reduced pressure and the crude material purified by flash chromatography (eluting with: cyclohexaneacetone 85\15) to give the title compound (1.51 g, % pure by NMR analysis).
m\z ([MH]+)=770.
To a solution of example 11 (1.5 g) in anhydrous DCM (5.4 mL) trifluoroacetic acid (0.6 mL) was added under nitrogen atmosphere and the resulting mixture was stirred for 2 h at room temperature. The reaction mixture was diluted with EtOAc (15 mL) and concentrated under reduced pressure. The residue was dissolved in DCM (100 mL) and washed with a saturated NaHCO3 aqueous solution (50 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound (1.05 g).
m\z ([MH]+)=670.
1H-NMR (CDCl3) δ: 5.12 (dd, 1H), 4.32 (d, 1H), 4.21 (d, 1H), 4.05 (m, 1H), 3.85 (q, 1H), 3.58 (m, 1H), 3.25 (dd, 1H), 3.20-3.0 (m, 4H), 3.05 (m, 2H), 2.67 (m, 1H), 2.68 (s, 3H), 2.61 (m, 1H), 2.40 (s, 6H), 2.32 (m, 1H), 1.91 (m, 1H), 1.78-1.68 (m, 3H), 1.60 (m, 1H), 1.47 (s, 3H), 1.38 (d, 3H), 1.32 (s, 3H), 1.30 (d, 3H), 1.29 (m, 1H), 1.27 (d, 3H), 1.15 (d, 3H), 1.11 (d, 3H), 0.88 (t, 3H).
To a solution of intermediate 25 (0.035 g) in anhydrous DCM (1.5 mL), EDC (0.041 g) and DMSO (0.041 mL) were added at 0° C. under nitrogen atmosphere. After stirring at 0° C. for 15 min, pyridinium trifluoroacetate (0.042 g) was added. The reaction mixture was stirred for 3 h at room temperature, then quenched with water (3 mL) and extracted with DCM (5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by preparative TLC (eluting with: DCM\MeOH 95\5) to give the title compound (0.016 g).
m\z ([MH]+)=838.
A solution of example 14 (0.016 g) in MeOH (1.5 mL) was stirred for 24 h then concentrated under reduced pressure to give the title compound (0.009 g).
1H-NMR (CDCl3) δ: 9.50 (d, 1H), 8.79 (d, 1H), 8.16 (d, 1H), 7.94 (d, 1H), 7.84 (t, NH), 7.81 (t, 1H), 7.61 (t, 1H), 4.90 (dd, 1H), 4.27 (d, 1H), 4.19 (d, 1H), 4.01 (m, 2H), 3.84 (d, 1H), 3.52 (m, 3H), 3.14 (m, 3H), 2.70 (m, 1H), 2.53 (s, 3H), 2.54 (m, 1H), 2.43 (m, 1H), 2.27 (s, 6H), 1.95 (m, 1H), 1.84 (d, 1H), 1.75 (m, 1H), 1.64 (m, 1H), 1.60 (m, 1H), 1.56 (s, 3H), 1.38 (d, 3H), 1.30 (d, 3H), 1.26 (s, 3H), 1.22 (m, 1H), 1.22 (d, 3×3H), 0.88 (t, 3H).
To a solution of intermediate 26 (0.040 g) in anhydrous DCM (3 mL) Dess-Martin periodinane (0.030 g) was added portionwise at room temperature within 5 h. The reaction was quenched with a Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 2 mL), stirred for 1 h then extracted with DCM (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.029 g).
TLC: DCM\MeOH 90\10 (Rf=0.42).
A solution of example 16 (0.029 g) in MeOH (2 mL) was stirred for 24 h then concentrated under reduced pressure to give the title compound (0.023 g).
1H-NMR (CDCl3) δ: 8.98 (d, 1H), 8.40 (dd, 1H), 8.10 (dd, 1H), 7.61 (d, 1H), 7.35 (d, 1H), 7.29 (m, 1H), 6.48 (t, NH), 4.89 (dd, 1H), 4.30 (d, 2H), 4.21 (d, 1H), 4.01 (m, 1H), 3.83 (q, 1H), 3.73 (m, 1H), 3.55 (m, 1H), 3.34 (m, 1H), 3.19 (m, 1H), 3.08 (m, 2H), 2.65 (s, 3H), 2.58 (m, 1H), 2.47 (m, 1H), 2.34 (m, 1H), 2.28 (s, 6H), 2.3-2.1 (m, 4H), 1.91 (m, 1H), 1.81 (m, 1H), 1.80-1.54 (m, 3H), 1.51 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.32 (s, 3H), 1.30 (d, 3H), 1.25 (d, 3H), 1.12 (d, 3H), 1.15(d, 3H), 1.12 (d, 3H), 0.87 (t, 3H).
To a solution of intermediate 27 (0.039 g) in anhydrous DCM (3 mL) under nitrogen atmosphere Dess Martin periodinane (0.030 g) was added portionwise at room temperature within 5 h. The reaction was quenched with a Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 7 mL), stirred for 1 h and then extracted with DCM (15 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.026 g).
TLC: DCM\MeOH 90\10 (Rf=0.35).
A solution of example 18 (0.024 g) in MeOH (1 mL) was stirred for 24 h then concentrated under reduced pressure to give the title compound (0.020 g).
1H-NMR (CDCl3) δ: 8.99 (d, 1H), 8.45 (dd, 1H), 8.08 (dd, 1H), 7.58 (d, 1H), 7.35 (d, 1H), 7.29 (m, 1H), 6.67 (t, NH), 4.80 (dd, 1H), 4.36 (m, 2H), 4.30 (d, 1H), 4.19 (d, 1H), 3.83 (m, 2H), 3.61 (m, 1H), 3.55 (m, 1H), 3.25 (m, 2H), 3.04 (m, 2H), 2.70 (m, 2H), 2.62 (s, 3H), 2.58 (m, 2H), 2.33 (s, 6H), 2.32 (d, 1H), 1.85 (m, 1H), 1.80 (m, 1H), 1.70 (m, 2H), 1.56 (m, 1H), 1.48 (s, 3H), 1.45 (d, 3H), 1.33 (s, 3H), 1.21 (d, 3H), 1.35 (m, 1H), 1.18 (d, 3H), 1.04 (d, 3H), 0.95 (t, 3H), 0.91 (d, 3H).
To a solution of intermediate 28 (0.018 g) in anhydrous DCM (1.5 mL) under nitrogen atmosphere Dess-Martin periodinane (0.020 g) was added portionwise at room temperature within 5 h. The reaction was quenched with a Na2S2O3solution (5% in a saturated NaHCO3 aqueous solution, 2 mL), stirred for 1 h then extracted with DCM (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by preparative TLC (eluting with: DCM\MeOH 10\1) and the recovered silica gel was stirred overnight in MeOH (1 mL). The mixture was filtered and solvent evaporation under reduced pressure gave the title compound (0.009 g).
1H-NMR (CDCl3) δ: 9.02 (d, 1H), 8.49 (m, 1H), 8.11 (m, 1H), 7.65 (d, 1H), 7.43 (d, 1H), 7.30 (m, 1H), 6.58 (t, NH), 4.89 (dd, 1H), 4.70 (dd, 2H), 4.29 (d, 1H), 4.20 (d, 1H), 3.83 (m, 2H), 3.69 (m, 1H), 3.51 (m, 1H), 3.29 (m, 1H), 3.17 (m, 1H), 3.04 (m, 2H), 2.60 (s, 3H), 2.54 (m, 1H), 2.44 (m, 1H), 2.31 (m, 1H), 2.27 (s, 6H), 1.91 (m, 1H), 1.80 (m, 1.68 (m, 2H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.29 (s, 3H), 1.29 (d, 3H), 1.22 (m, 1H), 1.25 (d, 3H), 1.12 (d, 3H), 1.10 (d, 3H), 0.87 (t, 3H).
To a solution of intermediate 29 (0.020 g) in anhydrous DCM (2 mL) under nitrogen atmosphere Dess-Martin periodinane (0.017 g) was added portionwise at room temperature within 1 h. After stirring for 5 h the reaction was quenched with a solution Na2S2O3 (5% in a saturated NaHCO3 aqueous solution, 2 mL), stirred for 1 h then extracted with DCM (3×5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Evaporation of the solvent gave the title compound (0.010 g).
1H-NMR (CDCl3) δ: 4.39 (m, 1H), 3.76 (m, 1H), 3.52 (m, 1H), 3.39 (m, 1H), 3.15 (m, 1H), 3.04 (m, 1H), 2.41 (m, 1H), 2.36 (m, 1H), 2.32 (m, 1H), 2.10 (m, 1H), 2.02 (m, 1H), 1.12 (d, 3H).
To a solution of intermediate 30 (0.068 g) in anhydrous DCM (5 mL) Dess-Martin periodinane (0.064 g) was added under nitrogen atmosphere. The mixture was stirred at room temperature for 6 h. The reaction was quenched with a solution Na2S2O3 (5% in a saturated NaHCO3 aqueous solution, 5 mL), stirred for 30 min and then extracted with DCM (3×4 mL). The organic phase was washed with brine (5 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was dissolved in MeOH (5 mL) and stirred overnight. Solvent evaporation under reduced pressure gave the title compound (0.053 g).
1H-NMR (CDCl3) δ: 8.70 (s, 1H), 8.05 (d, 1H), 8.02 (d, 1H), 7.71 (td, 1H), 7.64 (td, 1H), 7.59 (bt, 1H), 4.11 (d, 1H), 4.03 (m, 1H), 3.34 (bm, 1H), 2.98 (m, 1H), 1.01 (d, 3H).
To a solution of intermediate 31 (0.035 g) in anhydrous DCM (2.5 mL) Dess-Martin periodinane (0.033 g) was added under nitrogen atmosphere. The mixture was stirred at room temperature for 18 h. The reaction was quenched with a Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 2 mL), stirred for 30 min and then extracted with DCM (3×4 mL). The organic phase was washed with brine (3 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was dissolved in anhydrous MeOH (2 mL) and stirred overnight. The solvent was evaporated under reduced pressure and the crude material purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give the title compound (0.011 g).
1H-NMR (CDCl3) δ: 8.50 (s, 1H), 8.00 (d, 1H), 7.98 (d, 1H), 7.70 (t, 1H), 7.61 (t, 1H), 6.69 (bt, 1H), 3.83 (m, 1H), 3.68 (m, 1H), 3.64 (m, 1H), 3.34 (m, 1H), 3.06 (m, 1H), 2.77 (m, 1H), 2.34 (m, 1H), 1.11 (d, 3H).
To a solution of intermediate 32 (0.100 g) in anhydrous DCM (4 mL) Dess Martin periodinane (0.110 g) was added portionwise within 3 h under nitrogen atmosphere. The mixture was stirred at room temperature for 3 h. The reaction was quenched with a Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 2 mL), stirred for 45 min and then extracted with DCM (3×4 mL). The organic phase was washed with brine (3 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by preparative TLC (eluting with: DCM\MeOH 95\5), the recovered silica gel stirred for 18 h in MeOH (5 mL), the mixture filtered and then solvent evaporation under reduced pressure gave the title compound (0.015 g).
m\z ([MH]+)=780.
To a solution of example 24 (0.015 g) in anhydrous MeOH (6 mL) palladium (10wt. % on carbon powder, 0.005 g) was added and the mixture stirred under hydrogen atmosphere (6 atm) for 20 h. Filtration through a celite pad eluting with MeOH (20 mL) and purification by flash chromatography (eluting with DCM\MeOH from 100\0 to 95\5) and by preparative TLC (eluting with: DCM\MeOH\NH4OH 87.5\12.5\0.5) gave the title compound (0.002 g).
1H-NMR (CDCl3) δ: 8.87 (d, 1H), 8.10 (d, 2H), 7.61 (d, 1H), 7.70 (m, 1H), 7.57 (m, 1H), 4.38 (d, 1H), 4.26 (d, 1H), 3.55 (m, 1H), 3.28-3.00 (m, 2H).
To a solution of example 6 (0.020 g) in anhydrous DMF (2 mL) under nitrogen atmosphere intermediate 48 (0.009 g), HATU (0.013 g) and DIPEA (0.013 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (15 mL) and washed with water (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give a compound that was dissolved in MeOH (1 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.009 g).
1H-NMR (CDCl3) δ: 9.04 (d, 1H), 8.44 (dd, 1H), 8.13 (m, 1H), 7.61 (m, 1H), 7.41 (d, 1H), 6.73 (bd, 1H), 5.08 (dd, 1H), 4.94 (m, 1H), 4.45 (m, 1H), 4.26 (m, 1H), 4.25 (d, 1H), 4.08 (m, 3H), 3.80 (q, 1H), 3.51 (m, 1H), 3.15 (m, 1H), 3.04 (m, 1H), 2.98 (m, 1H), 2.65 (m, 1H), 2.60 (m, 1H), 2.52 (m, 1H), 2.44 (m, 1H), 2.39 (s, 3H), 2.34 (m, 1H), 2.26 (s, 6H), 1.90 (m, 1H), 1.75 (m, 1H), 1.65-1.55 (m, 3H), 1.48 (s, 3H), 1.37 (d, 3H), 1.27 (d, 3H), 1.22 (d, 3H), 1.25 (m, 1H), 1.17 (s, 3H), 1.16 (d, 3H), 1.12 (d, 3H), 0.87 (t, 3H).
To a solution of example 6 (0.020 g) in anhydrous DMF (2 mL) under nitrogen atmosphere intermediate 49 (0.009 g), HATU (0.013 g) and DIPEA (0.013 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (15 mL) and washed with water (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5, 90\10) and the (21R) and (21S) isomers were isolated. Each isomer was dissolved in MeOH (1 mL) and stirred at room temperature overnight. Evaporation of the solvent under reduced pressure gave the title compound 27 (0.003 g) and the title compound 28 (0.006 g).
1H-NMR (CDCl3) (example 27): 8.98 (d, 1H), 8.89 (bd, 1H), 8.46 (d, 1H), 8.08 (d, 1H), 7.61 (d, 1H), 7.41 (d, 1H), 7.28 (m, 1H), 5.33 (dd, 1H), 5.22 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.14 (m, 2H), 3.88 (q, 1H), 3.56 (m, 1H), 3.27 (m, 1H), 3.17 (dd, 1H), 3.06 (m, 1H), 2.99 (m, 1H), 2.66 (m, 1H), 2.57 (s, 3H), 2.46 (m, 1H), 2.40 (m, 2H), 2.27 (s, 6H), 2.21 (m, 2H), 1.86 (m, 1H), 1.84 (m, 1H), 1.70-1.50 (m, 3H), 1.54 (s, 3H), 1.36 (d, 3H), 1.33 (d, 3H), 1.29 (s, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.17 (d, 3H), 1.08 (d, 3H), 0.90 (t, 3H).
1H-NMR (CDCl3) (example 28): 9.02 (d, 1H), 8.46 (dd, 1H), 8.11 (m, 1H), 7.61(m, 1H), 7.37 (d, 1H), 7.31 (m, 1H), 6.47 (bd, 1H), 5.26 (dd, 1H), 4.81 (bt, 1H), 4.29 (d, 1H), 4.13 (m, 3H), 3.81 (q, 1H), 3.54 (m, 1H), 3.17 (m, 1H), 3.08-3.04 (m, 2H), 2.53 (s, 3H), 2.56 (m, 1H), 2.45 (m, 1H), 2.36 (m, 1H), 2.27 (s, 6H), 2.17-2.13 (m, 4H), 2.00 (m, 1H), 1.80 (m, 1H), 1.68 (m, 1H), 1.60 (m, 1H), 1.51 (s, 3H), 1.40 (d, 3H), 1.29 (s, 3H), 1.30 (d, 3H), 1.25 (m, 1H), 1.22 (d, 3H), 1.19 (d, 3H), 1.16 (d, 3H), 0.91 (t, 3H).
To a solution of example 6 (0.100 g) in anhydrous DMF (5 mL) under nitrogen atmosphere intermediate 50 (0.048 g), HATU (0.063 g) and DIPEA (0.061 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (30 mL) and washed with water (25 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5, 90\10) to give a compound that was dissolved in MeOH (5 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.090 g).
1H-NMR (CDCl3) δ: 8.98 (d, 1H), 8.45 (d, 1H), 8.09 (d, 1H), 7.55 (s, 1H), 7.34 (s, 1H), 7.29 (m, 1H), 6.45 (bd, 1H), 5.19 (dd, 1H), 4.82 (m, 1H), 4.29 (d, 1H), 4.12 (d, 1H), 3.98 (m, 2H), 3.80 (q, 1H), 3.55 (m, 1H), 3.17 (m, 1H), 3.06 (m, 2H), 2.55 (s+m, 1H+3H), 2.45 (m, 1H), 2.35 (m, 1H), 2.27 (s, 6H), 2.21 (m, 2H), 1.98 (m, 1H), 1.88 (m, 2H), 1.80 (m, 2H), 1.60-1.50 (m, 3H), 1.49 (s, 3H), 1.39 (d, 3H), 1.35 (m, 1H), 1.30 (d+s, 3H+3H), 1.23 (d, 3H), 1.16 (d, 3H+3H), 0.89 (t, 3H).
To a solution of example 6 (0.100 g) in anhydrous DMF (8 mL) under nitrogen atmosphere a solution of intermediate 47 (0.040 g) in anhydrous DMF (2 mL), HATU (0.057 g) and DIPEA (0.060 mL) were sequentially added. The mixture was stirred at room temperature for 8 h. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (30 mL) and washed with water (2×10 mL). The aqueous phase was extracted again with DCM (20 mL). The organic layers were collected, dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 95\5 to 92\8) to give the title compound 30 (0.008 g) and the title compound 31 (0.021 g).
TLC: DCM\MeOH 95\5 (Rf (example 30)=0.53)
TLC: DCM\MeOH 95\5 (Rf (example 31)=0.47).
A solution of example 30 (0.007 g) in MeOH (5 mL) was stirred at room temperature for 48 h. After evaporating the solvent under reduced pressure, the crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 80\20) to give the title compound (0.003 g).
1H-NMR (CDCl3) δ: 9.19 (d, 1H), 8.98 (d, 1H), 8.47 (dd, 1H), 8.06 (d, 1H), 7.70 (d, 1H), 7.42 (d, 1H), 7.29 (m, 1H), 5.24 (d, 1H), 4.80 (dd, 1H), 3.18 (m, 1H), 2.94 (d, 1H), 0.99 (d, 3H).
A solution of example 31 (0.021 g) in MeOH (5 mL) was stirred at room temperature for 48 h. After evaporating the solvent under reduced pressure, the crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 80\20) to give the title compound (0.006 g).
1H-NMR (CDCl3) δ: 8.99 (d, 1H), 8.47 (d, 1H), 8.09 (d, 1H), 7.58 (s, 1H), 7.41 (s, 1H), 7.30 (m, 1H), 6.73 (d, 1H), 4.88 (dd, 1H), 4.67 (s, 1H), 3.08 (m, 1H), 2.41 (d, 1H), 1.16 (d, 3H).
To a solution of example 6 (0.100 g) in anhydrous DMF (8 mL) under nitrogen atmosphere a solution of quinoline-4-carboxylic acid (0.026 g) in anhydrous DMF (2 mL), HATU (0.057 g) and DIPEA (0.060 mL) were added. The mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (30 mL) and washed with a saturated NaHCO3 aqueous solution (2×10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 94\6). The obtained compound was dissolved in MeOH (10 mL) and stirred overnight. After solvent evaporation, purification by flash chromatography (eluting with: DCM\MeOH 90\10) gave the title compound (0.032 g).
1H-NMR (CDCl3) δ: 8.96 (d, 1H), 8.45 (d, 1H), 8.13 (d, 1H), 7.76 (t, 1H), 7.65 (t, 1H), 7.52 (d, 1H), 7.02 (d, 1H), 5.08 (dd, 1H), 3.17 (m, 1H), 2.60 (m, 1H), 1.28 (d, 3H).
To a solution of example 6 (0.100 g) in anhydrous DMF (8 mL) under nitrogen atmosphere 3-quinolin-4-yl-propionic acid (0.030 g), HATU (0.057 g) and DIPEA (0.060 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (30 mL) and washed with water (25 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 94\6) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.022 g).
1H-NMR (CDCl3) δ: 8.82 (d, 1H), 8.10 (d, 1H), 8.07 (d, 1H), 7.71 (t, 1H), 7.61 (t, 1H), 7.26 (d, 1H), 6.50 (bd, 1H), 5.14 (dd, 1H), 4.92 (dd, 1H), 4.28 (d, 1H), 4.12 (d, 1H), 3.80 (q, 1H), 3.54 (m, 1H), 3.46 (m, 2H), 3.16 (m, 1H), 3.08 (m, 1H), 3.04 (m, 1H), 2.60 (m, 2H), 2.49 (s, 3H), 2.45 (m, 1H), 2.39 (m, 1H), 2.26 (s, 6H), 2.00 (m, 1H), 1.80 (m, 2H), 1.72 (m, 1H), 1.65 (m, 1H), 1.60 (m, 1H), 1.50 (s, 3H), 1.39 (d, 3H), 1.29 (d, 3H), 1.28 (s, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.18 (d, 3H), 1.14 (d, 3H), 0.91 (t, 3H).
To a solution of example 6 (0.075 g) in anhydrous DMF (6 mL) under nitrogen atmosphere intermediate 60 (0.027 g), HATU (0.043 g) and DIPEA (0.047 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (20 mL) and washed with a saturated NaHCO3 aqueous solution (15 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 97\3 to 95\5) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.039 g).
1H-NMR (CDCl3) δ: 8.81 (d, 1H), 8.11 (d, 1H), 8.09 (d, 1H), 7.69 (t, 1H), 7.58 (t, 1H), 7,26 (m, 1H), 6.47 (bd, 1H), 5.20 (dd, 1H), 4.88 (dd, 1H), 4.30 (d, 1H), 4.13 (s, 1H), 4.13 (m, 2H), 3.80 (q, 1H), 3.53 (m, 1H), 3.19-3.00 (m, 5H), 2.54 (s, 3H), 2.61-2.41 (m, 3H), 2.29 (s+m, 2H+6H), 2.11 (m, 2H), 2.00 (m, 1H), 1.80-1.60 (m, 7H), 1.39 (d, 3), 1.29 (s+d, 3H+3H), 1.24 (m, 4H), 1.18 (d, 3H), 1.14 (d, 3H), 0.91 (t, 3H).
To a solution of example 6 (0.100 g) in anhydrous DMF (6 mL) under nitrogen atmosphere intermediate 62 (0.027 g), HATU (0.057 g) and DIPEA (0.057 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (20 mL) and washed with water (15 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.024 g).
1H-NMR (CDCl3) δ: 8.81 (d, 1H), 8.11 (d, 1H), 8.04 (m, 1H), 7.70 (m, 1H), 7.58 (m, 1H), 7.27 (m, 1H), 6.42 (bd, 1H), 5.18 (dd, 1H), 4.85 (dd, 1H), 4.30 (m, 1H), 4.13 (m, 1H), 3.79 (q, 1H), 3.54 (m, 1H), 3.17 (m, 1H), 3.08 (m, 4H), 2.58 (m, 1H), 2.55 (s, 3H), 2.45 (m, 1H), 2.40 (m, 1H), 2.27 (s, 6H), 2.23 (m, 2H), 1.98 (m, 1H), 1.81 (m, 4H), 1.75 (m, 1H), 1.70 (m, 1H), 1.65 (m, 1H), 1.60 (m, 1H), 1.50 (s, 3H), 1.40 (d, 3H), 1.30 (d, 3H), 1.28 (s, 3H), 1.24 (d, 3H), 1.20 (m, 1H), 1.18 (d, 3H), 1.14 (d, 3H), 0.89 (t, 3H).
To a solution of example 7 (0.050 g) in anhydrous DMF (4 mL) under nitrogen atmosphere a solution of intermediate 65 (0.018 g) in anhydrous DMF (1 mL), HATU (0.013 g) and DIPEA (0.013 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (15 mL) and washed with a saturated NaHCO3 aqueous solution (10 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Evaporation of the solvent under reduced pressure gave the title compound (0.031 g)
1H-NMR (CDCl3) δ: 7.78 (d, 2H), 7.47 (d, 1H), 7.31 (t, 2H), 7.27 (d, 1H), 7.16 (t, 1H), 6.64 (d, 1H), 5.04 (dd, 1H), 4.99 (m, 1H), 4.20 (m, 1H), 3.01 (m, 1H), 2.60-2.50 (m, 2H), 2.33 (dd, 1H), 1.12 (d, 3H).
To a suspension of intermediate 67 (0.022 g) in anhydrous DMF (1.6 mL) HATU (0.037 g) and DIPEA (0.015 mL) were sequentially added. The mixture was stirred under nitrogen atmosphere for 30 min then example 7 (0.050 g) was added. After stirring at room temperature overnight the reaction mixture was diluted with DCM (3.5 mL), washed with a 5% NaHCO3 aqueous solution (3 mL) while ice-cooling and the aqueous phase extracted with DCM (2.5 mL). The collected organic extracts were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (10 mL) and stirred at room temperature overnight. After solvent evaporation under reduced pressure the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 94\6\0.5) to give the title compound (0.010 g).
1H-NMR (DMSO) δ: 8.62 (d, 1H), 7.71 (d, 2H), 7.58 (d, 1H), 7.49 (d, 1H), 7.33 (t, 2H), 7.17 (t, 1H), 4.69 (d, 1H), 4.62 (d, 1H), 4.59 (d, 1H), 3.19 (m, 1H), 2.59 (m, 1H), 1.10 (d, 3H).
To a solution of example 7 (0.034 g) in anhydrous DMF (4 mL) under nitrogen atmosphere a solution of intermediate 69 (0.012 g) in anhydrous DMF (1 mL), HATU (0.019 g) and DIPEA (0.020 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give a compound that was dissolved in MeOH (5 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.020 g).
1H-NMR (CDCl3) δ: 7.46 (d, 1H), 7.31 (dd, 1H), 7.20 (d, 1H), 7.14 (dd, 1H), 7.00 (dd, 1H), 6.68 (d, 1H), 5.08 (dd, 1H), 4.40 (m, 1H), 4.20 (m, 1H), 3.05 (m, 1H), 2.68 (m, 1H), 2.50 (m, 1H), 2.38 (dd, 1H), 1.16 (d, 3H).
To a solution of example 7 (0.050 g) in anhydrous DMF (4 mL) under nitrogen atmosphere a solution of intermediate 70 (0.017 g) in anhydrous DMF (1mL), HATU (0.029 g) and DIPEA (0.030 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) to give a compound that was dissolved in MeOH (3 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure and purification by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) gave the title compound (0.007 g).
m\z ([MH]+)=832.
To a solution of example 6 (0.050 g) in anhydrous DMF (4 mL) under nitrogen atmosphere (2S)-tert-butoxycarbonylamino-3-(1H-indol-3-yl)-propionic acid (0.027 g), HATU (0.031 g) and DIPEA (0.031 mL) were sequentally added. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2 to 97\3) to give the title compound (0.043 g).
m\z ([MH]+)=955.
A solution of example 42 (0.013 g) in MeOH (2 mL) was stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.010 g).
m\z ([MH]+)=913.
To a solution of example 42 (0.025 g) in anhydrous DCM (0.5 mL) cooled to 0° C. trifluoroacetic acid (0.1 mL) was added. After removing the ice-bath, the reaction mixture was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2 mL). The aqueous phase was extracted with DCM (3×3 mL). The collected organic extracts were dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 97\3) to give a compound that was dissolved in MeOH (1 mL) and the solution was stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.004 g).
1H-NMR (CDCl3) δ: 8.12 (bs, 1H), 8.06 (d, 1H), 7.65 (d, 1H), 7.36 (d, 1H), 7.19 (t, 1H), 7.16 (d, 1H), 7.13 (t, 1H), 5.35 (dd, 1H), 4,75 (dd, 1H), 3.70 (dd, 1H), 3.34 (dd, 1H), 3.09-3.04 (m, 3H), 2.46-2.44 (m, 2H), 1.20 (d, 3H).
m\z ([MH]+)=813.
To a solution of (quinoxalin-2-ylsulfanyl)-acetic acid (0.095 g) in anhydrous DMF (8.3 mL) under nitrogen atmosphere HATU (0.164 g) and DIPEA (0.089 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.262 g) was added. The reaction mixture was stirred at room temperature for 20 h then it was diluted with DCM (30 mL) and washed with a 5% NaHCO3 aqueous solution (20 mL). The aqueous phase was extracted with DCM (25 mL), the collected organic layers were washed with a 5% NaHCO3 aqueous solution (20 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (10 mL) and stirred at room temperature overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0 to 93\7\1) to give the title compound (0.199 g).
1H-NMR (CDCl3) δ: 8.67 (s, 1H), 8.19 (dd, 1H), 8.04 (dd, 1H), 7.90 (d, 1H), 7.74 (dt, 1H), 7.66 (dt, 1H), 5.20 (dd, 1H), 4.67 (dd, 1H), 4.30 (d, 1H), 4.06 (m+d, 1H+1H), 3.95 (d, 1H), 3.73 (q, 1H), 3.52 (m, 2H), 3.16 (dd, 1H), 3.06-3.0 (m, 2H), 2.52 (s, 3H), 2.50-2.40 (m, 2H), 2.42 (dd, 1H), 2.27 (s, 6H), 1.92 (m, 1H), 1.81-1.65 (m, 2H), 1.66 (m, 1H), 1.51 (m, 1H), 1.47 (s, 3H), 1.36 (d, 3H), 1.27 (d, 3H), 1.24 (m, 1H), 1.24 (d, 3H), 1.23 (s, 3H), 1.12 (d, 3H), 1.06 (d, 3H), 0.81 (t, 3H).
To a solution of (quinoxalin-2-ylsulfanyl)-acetic acid (0.395 g) in anhydrous DMF (5 mL) under nitrogen atmosphere and HATU (0.682 g) and DIPEA (0.365 mL) were added at room temperature. The reaction mixture was stirred at room temperature for 30 min then a solution of example 7 (1.0 g) in anhydrous DMF (3 mL) was added. The reaction mixture was stirred at room temperature for 3 h then it was poured into a 5% NaHCO3 aqueous solution (30 mL) and the solution extracted with Et2O (2×30 mL). The collected organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (25 mL) and stirred at room temperature overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 93\7\0.2). Crystallisation from EtOAc gave the title compound (0.454 g, (21S) isomer 99% pure by NMR analysis).
1H-NMR (CDCl3) : 8.67 (s, 1H), 8.19 (dd, 1H), 8.04 (dd, 1H), 7.90 (d, 1H), 7.74 (dt, 1H), 7.66 (dt, 1H), 5.20 (dd, 1H), 4.67 (dd, 1H), 4.30 (d, 1H), 4.06 (m+d, 1H+1H), 3.95 (d, 1H), 3.73 (q, 1H), 3.52 (m, 2H), 3.16 (dd, 1H), 3.06-3.0 (m, 2H), 2.52 (s, 3H), 2.50-2.40 (m, 2H), 2.42 (dd, 1H), 2.27 (s, 6H), 1.92 (m, 1H), 1.81-1.65 (m, 2H), 1.66 (m, 1H), 1.51 (m, 1H), 1.47 (s, 3H), 1.36 (d, 3H), 1.27 (d, 3H), 1.24 (m, 1H), 1.24 (d, 3H), 1.23 (s, 3H), 1.12 (d, 3H), 1.06 (d, 3H), 0.81 (t, 3H).
To a solution of 4-(2,3-dihydro-benzo[1,4]dioxin-6-yl)4-oxo-butyric acid (0.1165 g) in anhydrous DMF (9.5 mL) under nitrogen atmosphere HATU (0.188 g) and DIPEA (0.102 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 7 (0.300 g) was added. The reaction mixture was stirred at room temperature for 20 h then it was diluted with DCM (30 mL) and washed with a 5% NaHCO3 aqueous solution (20 mL). The aqueous phase was extracted with DCM (25 mL), the collected organic layers were washed with a 5% NaHCO3 aqueous solution (20 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (10 mL) and stirred at room temperature overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH40H from 100\0\0 to 93\7\1) to give the title compound (0.170 g).
1H-NMR (CDCl3) δ: 7.52 (d, 2H), 6.89 (d, 1H), 6.73 (d, NH), 5.20 (dd, 1H), 4.84 (dd, 1H), 4.35-4.25 (m, 5H), 4.13 (d, 1H), 3.80 (q, 1H), 3.60-3.55 (m, 2H), 3.32-3.22 (m, 2H), 3.18 (dd, 1H), 3.10-3.00 (m, 2H), 2.65-2.55 (m, 2H), 2.60 (s, 3H), 2.44 (m, 2H), 2.27 (s, 6H), 2.27 (m, 1H), 1.99 (m, 1H), 1.80 (dd, 1H), 1.75 (m, 1H), 1.68 (m, 1H), 1.60 (m, 1H), 1.50 (s, 3H), 1.40 (d, 3H), 1.32 (s, 3H), 1.30 (d, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.19 (d, 3H), 1.17 (d, 3H), 0.93 (t, 3H).
To a solution of 4-(4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-4-oxo-butyric acid (0.013 g) in anhydrous DMF (0.850 mL) under nitrogen atmosphere and HATU (0.019 g) and DIPEA (0.010 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.028 g) was added. The reaction mixture was stirred at room temperature for 2.5 h then it was diluted with DCM (2 mL) and washed with a 5% NaHCO3 aqueous solution (2 mL). The aqueous phase was extracted with DCM (1.5 mL), the collected organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and stirred at room temperature for 24 h. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 92\8\0.2) to give the title compound (0.031 g).
1H-NMR (CDCl3) δ: 8.02 (s, 1H), 6.68 (d, NH), 5.13 (dd, 1H), 4.88 (dd, 1H), 4.32 (d, 1H), 4.14 (d, 1H), 3.81 (q, 1H), 3.55 (m, 1H), 3.28 (m, 2H), 3.21 (m, 1H), 3.09-3.03 (m, 4H), 2.62 (s, 3H), 2.63-2.57 (m, 6H), 2.46 (dd, 1H), 2.35 (s, 6H), 2.24 (m, 2H), 1.98 (m, 1H), 1.80-1.70 (m, 3H), 1.60 (m, 1H), 1.49 (s, 3H), 1.40 (d, 3H), 1.32 (s, 3H), 1.29 (d, 3H), 1.25 (m, 4H), 1.18 (d, 3H), 1.17 (d, 3H), 0.92 (t, 3H).
To a solution of 4-(4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-4-oxo-butyric acid (1.280 g) in anhydrous DMF (12 mL) under nitrogen atmosphere HATU (2.02 g) and DIPEA (1.06 mL) were added. The reaction mixture was stirred at room temperature for 30 min then a solution of example 7 (2.90 g) in anhydrous DMF (10 mL) was added. The reaction mixture was stirred at room temperature overnight then it was diluted with EtOAc (100 mL) and washed with a 5% NaHCO3 aqueous solution (2×50 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 98\2) to give a compound that was dissolved in MeOH (150 mL) and stirred overnight. After evaporating the solvent the residue was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (2.40 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 8.02 (s, 1H), 6.68 (d, NH), 5.13 (dd, 1H), 4.88 (dd, 1H), 4.32 (d, 1H), 4.14 (d, 1H), 3.81 (q, 1H), 3.55 (m, 1H), 3.28 (m, 2H), 3.21 (m, 1H), 3.09-3.03 (m, 4H), 2.62 (s, 3H), 2.63-2.57 (m, 6H), 2.46 (dd, 1H), 2.35 (s, 6H), 2.24 (m, 2H), 1.98 (m, 1H), 1.80 -1.70 (m, 3H), 1.60 (m, 1H), 1.49 (s, 3H), 1.40 (d, 3H), 1.32 (s, 3H), 1.29 (d, 3H), 1.25 (m, 4H), 1.18 (d, 3H), 1.17 (d, 3H), 0.92 (t, 3H).
To a solution of 4-(4-methoxy-3-nitro-phenyl)-4-oxo-butyric acid (0.136 g) in anhydrous DMF (9 mL) under nitrogen atmosphere HATU (0.205 g) and DIPEA (0.110 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.300 g) was added. The reaction mixture was stirred at room temperature for 2.5 h then it was diluted with DCM (21 mL) and washed with a 5% NaHCO3 aqueous solution (18 mL). The aqueous phase was extracted with DCM (15 mL), the collected organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (30 mL) and DCM (2 mL) and stirred at room temperature for 48 h. After evaporating the solvent the crude material was purified by LC (mobile phase: A\B 85\15 for 1 min, from 85\15 to 15\85 in 20 min; =255 nm) to give the title compound (0.149 g).
1H-NMR (CDCl3) δ: 8.47 (d, 1H), 8.19 (dd, 1H), 7.15 (d, 1H), 6.69 (d, NH), 5.18 (dd, 1H), 4.85 (dd, 1H), 4.31 (d, 1H), 4.14 (d, 1H), 4.04 (s, 3H), 3.80 (q, 1H), 3.54 (m, 1H), 3.32 (m, 2H), 3.18 (dd, 1H), 3.06-3.0 (m, 2H), 2.66 (m, 2H), 2.60 (s, 3H), 2.46 (m, 1H), 2.42 (dd, 1H), 2.28 (s, 6H), 1.98 (m, 1H), 1.81-1.55 (m, 4H), 1.49 (s, 3H), 1.40 (d, 3H), 1.32 (s, 3H), 1.30 (d, 3H), 1.25 (d, 3H), 1.25 (m, 1H), 1.18 (d, 3H), 1.17 (d, 3H), 0.81 (t, 3H).
To a solution of 4-(4-methoxy-3-nitro-phenyl)-4-oxo-butyric acid (2.782 g) in anhydrous DMF (36 mL) under nitrogen atmosphere HATU (1.230 g) and DIPEA (0.656 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 7 (1.8 g) was added. The reaction mixture was stirred at room temperature for 3 h then it was diluted with Et2O (200 mL) and washed with a 5% NaHCO3 aqueous solution (2×100 mL). The aqueous phase was extracted with Et2O (50 mL) and DCM (50 mL), the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 96\4). The obtained compound was dissolved in MeOH (100 mL) and stirred at room temperature overnight. After evaporating the solvent the product was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 96\4) to give the title compound (1.510 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 8.47 (d, 1H), 8.19 (dd, 1H), 7.15 (d, 1H), 6.69 (d, NH), 5.18 (dd, 1H), 4.85 (dd, 1H), 4.31 (d, 1H), 4.14 (d, 1H), 4.04 (s, —OCH3), 3.80 (q, 1H), 3.54 (m, 1H), 3.32 (m, 2H), 3.18 (dd, 1H), 3.06-3.0 (m, 2H), 2.66 (m, 2H), 2.60 (s, 3H), 2.46 (m, 1H), 2.42 (dd, 1H), 2.28 (s, 6H), 1.98 (m, 1H), 1.81-1.55 (m, 4H), 1.49 (s, 3H), 1.40 (d, 3H), 1.32 (s, 3H), 1.30 (d, 3H), 1.25 (d, 3H), 1.25 (m, 1H), 1.18 (d, 3H), 1.17 (d, 3H), 0.81 (t, 3H).
To a solution of 4-(2-hydroxy-4,5-dimethoxy-phenyl)-4-oxo-butyric acid (0.137 g) in anhydrous DMF (9 mL) under nitrogen atmosphere HATU (0.205 g) and DIPEA (0.110 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.300 g) was added. The reaction mixture was stirred at room temperature for 2.5 h then it was diluted with DCM (21 mL) and washed with a 5% NaHCO3 aqueous solution (18 mL). The aqueous phase was extracted with DCM (15 mL), the combined organic layers were dried over Na2SO4 and evaporated under reduced pressure.
The residue was dissolved in MeOH (10 mL) and stirred at room temperature for 24 h. After evaporating the solvent the crude material was purified by LC (mobile phase: A\B from 85\15 to 15\85 in 20 min; =237 nm) to give the title compound (0.120 g).
1H-NMR (CDCl3) δ: 12.49 (s, 1H), 7.14 (s, 1H), 6.71 (d, NH), 6.44 (s, 1H), 5.18 (dd, 1H), 4.88 (dd, 1H), 4.31 (d, 1H), 4.14 (d, 1H), 3.91 (s, 3H), 3.88 (s, 3H), 3.80 (q, 1H), 3.55 (m, 1H), 3.41-3.30 (m, 2H), 3.20 (dd, 1H), 3.11-3.0 (m, 2H), 2.70-2.60 (m, 2H), 2.61 (s, 3H), 2.52 (m, 1H), 2.43 (dd, 1H), 2.31 (s, 6H), 1.98-1.70 (m, 3H), 1.60 (m, 1H), 1.50 (s, 3H), 1.30 (d, 3H), 1.26 (d, 3H), 1.25 (m, 1H), 1.24 (d+d, 6H), 1.19 (d, 3H), 1.17 (d, 3H), 0.91 (t, 3H).
To a solution of 4-(3-hydroxy-4-methoxy-phenyl)-4-oxo-butyric acid (0.121 g) in anhydrous DMF (9 mL) under nitrogen atmosphere HATU (0.205 g) and DIPEA (0.110 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.300 g) was added. The reaction mixture was stirred at room temperature for 2.5 h then it was diluted with DCM (21 mL) and washed with a 5% NaHCO3 aqueous solution (18 mL). The aqueous phase was extracted with DCM (15 mL), the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (30 mL) and DCM (2 mL) and stirred at room temperature for 48 h. After evaporating the solvent the crude material was purified by LC (mobile phase: A\B 85\15 for 1 min, from 85\15 to 15\85 in 20 min; =230 nm) to give the title compound (0.020 g).
1H-NMR (CDCl3) δ: 7.57 (m, 2H), 6.89 (d, 1H), 6.71 (d, 1H), 5.88 (bs, 1H), 5.22 (dd, 1H), 4.81 (dd, 1H), 4.31 (d, 1H), 4.13 (d, 1H), 3.96 (s, 3H), 3.81 (q, 1H), 3.55 (m, 1H), 3.31 (m, 2H), 3.22 (m, 1H), 3.09-3.04 (m, 2H), 2.66-2.54 (m, 4H), 2.61 (s, 3H), 2.44 (dd, 1H), 2.34 (s, 6H), 2.00 (m, 1H), 1.81-1.70 (m, 3H), 1.6 (m, 1H), 1.50 (s, 3H), 1.41 (d, 3H), 1.31 (s, 3H), 1.29 (d, 3H), 1.25 (d, 3H), 1.25 (m, 1H), 1.19 (d, 3H), 1.17 (d, 3H), 0.93 (t, 3H).
To a solution of 4-(3-hydroxy-4-methoxy-phenyl)-4-oxo-butyric acid (0.273 g) in anhydrous DMF (3.5 mL), HOBT (0.165 g) and EDC (0.234 g) were added under nitrogen atmosphere. After stirring 5 min example 7 (0.627 g) was added, and the resulting mixture was stirred at room temperature for 3 h. The reaction mixture was diluted with DCM (15 mL) washed with a 5% NaHCO3 aqueous solution (10 mL). The organic layer was washed with brine (10 ml), dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 97\3 to 95\5) to give a compound that was dissolved in MeOH (10 mL) and stirred at room temperature overnight. After solvent evaporation the compound was purified by preparative LC (column Waters XTerra MS C18 (19×300 mm, 7 μm); flow rate=12 ml\min, A\B from 70\30 to 10\90 in 25 min; =230 nm) to give the title compound (0.165 g, (21S) isomer 94% pure by NMR analysis).
1H-NMR (CDCl3) δ: 7.57 (m, 2H), 6.89 (d, 1H), 6.71 (d, 1H), 5.88 (bs, 1H), 5.22 (dd, 1H), 4.81 (dd, 1H), 4.31 (d, 1H), 4.13 (d, 1H), 3.96 (s, 3H), 3.81 (q, 1H), 3.55 (m, 1H), 3.31 (m, 2H), 3.22 (m, 1H), 3.09-3.04 (m, 2H), 2.66-2.54 (m, 4H), 2.61 (s, 3H), 2.44 (dd, 1H), 2.34 (s, 6H), 2.00 (m, 1H), 1.81-1.70 (m, 3H), 1.6 (m, 1H), 1.50 (s, 3H), 1.41 (d, 3H), 1.31 (s, 3H), 1.29 (d, 3H), 1.25 (d, 3H), 1.25 (m, 1H), 1.19 (d, 3H), 1.17 (d, 3H), 0.93 (t, 3H).
To a solution of 4-(3,4-dimethoxy-phenyl)-4-oxo-butyric acid (0.128 g) in anhydrous DMF (9 mL) under a nitrogen atmosphere HATU (0.205 g) and DIPEA (0.110 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 6 (0.300 g) was added. The reaction mixture was stirred at room temperature for 2.5 h then it was diluted with DCM (21 mL) and washed with a 5% NaHCO3 aqueous solution (18 mL). The aqueous phase was extracted with DCM (15 mL), the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (30 mL) and stirred at room temperature for 24 h. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 92\8\0.2) to give the title compound (0.293 g).
1H-NMR (CDCl3) δ: 7.95 (d, NH), 7.70 (dd, 1H), 7.55 (d, 1H), 6.99 (d, 1H), 5.30 (dd, 1H), 5.30 (dd, 1H), 4.71 (m, 1H), 4.37 (d, 1H), 4.08 (d, 1H), 3.96-3.93 (m, 4H), 3.90 (q, 1H), 3.68 (m, 1H), 3.39 (dd, 1H), 3.34 (m, 2H), 3.19 (m, 1H), 3.14 (m, 1H), 2.75 (s, 6H), 2.66-2.61 (m, 6H), 2.49 (d, 1H), 1.99-1.96 (m, 2H), 1.84-1.71 (m, 2H), 1.64 (m, 1H), 1.56 (s, 3H), 1.41 (m+d, 1H+3H), 1.32 (d, 3H), 1.31 (s, 3H), 1.26 (d, 3H), 1.23 (d, 3H), 1.21 (d, 3H), 0.95 (t, 3H).
To a solution of 4-(3,4-dimethoxy-phenyl)-4-oxo-butyric acid (0.695 g) in anhydrous DMF (15 mL) under nitrogen atmosphere HATU (1.08 g) and DIPEA (0.586 mL) were added. The reaction mixture was stirred at room temperature for 30 min then example 7 (1.50 g) was added. The reaction mixture was stirred at room temperature for 18 h then it was diluted with DCM (35 mL) and washed with a 5% NaHCO3 aqueous solution (30 mL). The aqueous phase was extracted with DCM (25 mL), the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (50 mL) and stirred at room temperature for 18 h. After evaporating the solvent the crude material was crystallised from DCM to give the title compound (1.26 g, (21S) isomer 98% pure by NMR analysis).
1H-NMR (CDCl3) δ: 7.95 (d, NH), 7.70 (dd, 1H), 7.55 (d, 1H), 6.99 (d, 1H), 5.30 (dd, 1H), 5.30 (dd, 1H), 4.71 (m, 1H), 4.37 (d, 1H), 4.08 (d, 1H), 3.96-3.93 (m, 4H), 3.90 (q, 1H), 3.68 (m, 1H), 3.39 (dd, 1H), 3.34 (m, 2H), 3.19 (m, 1H), 3.14 (m, 1H), 2.75 (s, 6H), 2.66-2.61 (m, 6H), 2.49 (d, 1H), 1.99-1.96 (m, 2H), 1.84-1.71 (m, 2H), 1.64 (m, 1H), 1.56 (s, 3H), 1.41 (m+d, 1H+3H), 1.32 (d, 3H), 1.31 (s, 3H), 1.26 (d, 3H), 1.23 (d, 3H), 1.21 (d, 3H), 0.95 (t, 3H).
To a solution of 4-(4-hydroxy-3-methoxy-phenyl)-4-oxo-butyric acid (0.922 g) in anhydrous DMF (20 mL) HOBT (0.656 g) and EDC (0.933 g) were added. Then example 6 (2.5 g) was added, and the resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with DCM (75 mL) washed with a 1N HCl aqueous solution (30 mL) then with a saturated NaHCO3 aqueous solution (30 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 98\2) then dissolved in MeOH (10 mL) and stirred at room temperature overnight. Solvent evaporation gave the title compound (0.600 g).
1H-NMR (CDCl3) δ: 7.57 (dd, 1H), 7.50 (d, 1H), 6.92 (d, 1H), 6.89 (d, 1H), 6.05 (bs, 1H), 5.19 (dd, 1H), 4.80 (dd, 1H), 4.28 (d, 1H), 4.10 (d, 1H), 3.93 (s, 3H), 3.77 (q, 1H), 3.58 (bs, 1H), 3.51 (m, 1H), 3.30 (m, 2H), 3.17 (dd, 1H), 3.06 (m, 1H), 3.02 (m, 1H), 2.6 (m, 3H), 2.57 (s, 3H), 2.48 (bm, 1H), 2.41 (dd, 1H), 2.28 (bs, 6H), 1.95 (m, 1H), 1.81-1.50 (m, 4H), 1.47 (s, 3H), 1.37 (d, 3H), 1.28 (s, 3H), 1.27 (d, 4H), 1.22 (d, 3H), 1.17 (d, 3H), 1.15 (d, 3H), 0.90 (t, 3H).
To a solution of 4-(4-hydroxy-3-methoxy-phenyl)-4-oxo-butyric acid (0.720 g) in anhydrous DMF (12 mL) HOBT (0.259 g) and EDC (0.368 g) were added under nitrogen atmosphere. Then example 7 (1.65 g) was added, and the resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with DCM (50 mL) washed with a 1N HCl aqueous solution (20 mL) then with a saturated NaHCO3 aqueous solution (20 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 98\2) then dissolved in MeOH (10 mL) and stirred at room temperature overnight. Solvent evaporation gave the title compound (0.360 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 7.57 (dd, 1H), 7.50 (d, 1H), 6.92 (d, 1H), 6.89 (d, 1H), 6.05 (bs, 1H), 5.19 (dd, 1H), 4.80 (dd, 1H), 4.28 (d, 1H), 4.10 (d, 1H), 3.93 (s, 3H), 3.77 (q, 1H), 3.58 (bs, 1H), 3.51 (m, 1H), 3.30 (m, 2H), 3.17 (dd, 1H), 3.06 (m, 1H), 3.02 (m, 1H), 2.6 (m, 3H), 2.57 (s, 3H), 2.48 (bm, 1H), 2.41 (dd, 1H), 2.28 (bs, 6H), 1.95 (m, 1H), 1.81-1.50 (m, 4H), 1.47 (s, 3H), 1.37 (d, 3H), 1.28 (s, 3H), 1.27 (d, 4H), 1.22 (d, 3H), 1.17 (d, 3H), 1.15 (d, 3H), 0.90 (t, 3H).
To a solution of intermediate 72 (0.032 g) in anhydrous DMF (2 mL) HATU (0.048 g) and DIPEA (0.022 mL) were added under nitrogen atmosphere. After stirring for 45 min example 7 (0.070 g) was added and the mixture stirred overnight. Water (5 mL) was added and the solution extracted with DCM (2×10 mL). The organic phase was washed with brine (5 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and stirred overnight. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 94\6) to give the title compound (0.060 g).
m\z ([MH]+)=859
1H-NMR (CDCl3) δ: 8.09 (d, 1H), 7.78 (d, 1H), 7.81 (d, 1H), 7.56 (t, 1H), 7.53 (t, 1H), 4.65 (t, 1H), 4.15 (s, 3H), 4.06 (d, 1H), 3.90 (d, 1H), 3.03 (m, 1H), 2.42 (m, 1H), 1.10 (d, 3H).
To a solution of intermediate 74 (0.038 g) in anhydrous DMF (1.5 mL) HATU (0.072 g) and DIPEA (0.033 mL) were added under nitrogen atmosphere. After stirring for 45 min example 7 (0.090 g) was added and the mixture stirred overnight. Water (8 mL) was added and the solution extracted with DCM (2×5 mL). Collected organic phases were washed with a saturated NaHCO3 aqueous solution (10 mL), brine (10 mL), then dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and stirred overnight. Purification of the crude material by flash chromatography (eluting with: DCM\MeOH 95\5) gave the title compound (0.047 g).
m\z ([MH]+)=813
1H-NMR (CDCl3) δ: 8.35 (s, 1H), 7.86 (d, 1H), 7.57 (td, 1H), 7.36 (m, 2H), 7.26 (d, 1H), 5.10 (dd, 1H), 5.02 (d, 1H), 4.87 (d, 1H), 3.01 (m, 1H), 2.34 (m, 1H), 1.11 (d, 3H).
To a solution of example 51 (0.070 g) in anhydrous MeOH (5 mL) palladium (10 wt. % on carbon powder, 0.050 g) was added and the mixture stirred under hydrogen atmosphere (1 atm) for 1 hr. Filtration through a silica pad eluting with MeOH and purification by preparative TLC (eluting with: DCM\MeOH 90\10) gave the title compound (0.008 g).
1H-NMR (CDCl3) δ: 8.47 (d, 1H), 8.19 (d, 1H), 7.15 (d, 1H), 6.68 (d, 1H), 4.82 (d, 1H), 4.04 (s, 3H), 3.32 (t, 2H), 3.07 (m, 1H), 2.70-2.60 (m, 2H), 2.39 (m, 1H), 2.08 (s, 3H), 1.11 (d, 3H)
To a solution of example 51 (0.050 g) in anhydrous MeOH (1 mL) hydroxylamine hydrochloride (0.050 g) and ammonium acetate (0.100 g) were added. The reaction mixture was stirred at 60° C. for 3 h. After solvent evaporation, the crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.015 g).
1H-NMR (CDCl3) δ: 8.17 (d, 1H), 7.82 (dd, 1H), 7.13 (d, 1H), 6.58 (bm, 1H), 4.80 (m, 1H), 3.00 (m, 1H), 3.10 (m, 2H), 2.50 (m, 2H), 2.34 (m, 1H), 1.14 (d, 3H).
To a solution of example 46 (0.050 g) in a 1\1 mixture of water\DCM (4 mL) magnesium monoperoxyphtalate (0.040 g) was added. After stirring for 24 h the aqueous phase was extracted with DCM (3×5 mL), the organic extracts washed with a 5% Na2S2O5 aqueous solution (5 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in THF (1 mL), polystyrene triphenylphosphine resin (0.050 g) was added and the mixture heated to 65° C. for 2 h. After cooling to room temperature, the mixture was filtered and concentrated to give the title compound (0.050 g).
1H-NMR (CDCl3) δ: 9.52 (s, 1H), 8.24 (m, 1H), 8.00-7.80 (m, 2H), 7.70 (bm, 1H), 4.83 (m, 1H), 4.49 (d, 1H), 4.04 (d, 1H), 3.06 (m, 1H), 2.50 (m, 1H), 1.15 (d, 3H).
A solution of example 6 (0.045 g) and 3-phenyl propionaldehyde (0.010 mL) in anhydrous MeOH (1 mL) was stirred at room temperature for 1 h under nitrogen atmosphere. Then sodium cyanoborohydride (0.007 g) and acetic acid (0.004 mL) were added. The mixture was stirred at room temperature for 1 h then the reaction was quenched with a saturated NaHCO3 aqueous solution (2 mL). After solvent evaporation, the aqueous phase was extracted with DCM (3×10 mL), the organic layer dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (1 mL) and stirred at room temperature overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM MeOH 95\5) to give the title compound (0.039 g).
1H-NMR (CDCl3) δ: 7.40-7.00 (m, 5H), 5.74 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.13 (s, 1H), 3.84 (q, 1H), 3.54 (m, 1H), 3.18 (m, 1H), 3.11 (m, 1H), 3.03 (m, 1H), 2.92-2.83 (m, 2H), 2.68 (s, 3H), 2.64 (m, 2H), 2.58 (m, 1H), 2.46 (m, 1H), 2.33 (m, 1H), 2.27 (s, 6H), 1.92 (m, 1H), 1.76 (m, 4H), 1.67 (m, 1H), 1.57 (m, 1H), 1.48 (s, 3H), 1.38 (d, 3H), 1.31 (d, 3H), 1.33 (s, 3H), 1.25 (d, 3H), 1.15 (d, 3H), 1.09 (d, 3H), 0.87 (t, 3H).
TLC: DCM\MeOH 10\1 (Rf=0.25).
A solution of example 6 (0.100 g) and intermediate 52 (0.040 g) in anhydrous DCM (4 mL) was stirred at room temperature for 6 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.150 mL) and acetic acid (0.009 mL) were added and the mixture stirred for 48 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and heated to reflux temperature for 48 h. After evaporating the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 90\10) to give the title compound (0.008 g).
1H-NMR (CDCl3) δ: 9.00 (d, 1H), 8.45 (d, 1H), 8.11 (d, 1H), 7.59 (d, 1H), 7.39 (d, 1H), 7.29 (m, 1H), 5.55 (dd, 1H), 4.29 (d, 1H), 4.24 (d, 1H), 4.13 (s, 1H), 4.20-4.0 (m, 2H), 3.86 (q, 1H), 3.55 (m, 1H), 3.19 (m, 1H), 3.09 (m, 1H), 3.05 (m, 1H), 2.93 (m, 2H), 2.67 (s, 3H), 2.52 (m, 1H), 2.49 (m, 1H), 2.32 (d, 1H), 2.27 (s, 6H), 2.02 (m, 1H), 1.94 (m, 2H), 1.80 (m, 2H), 1.65 (m, 1H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.33 (s, 3H), 1.31 (d, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.16 (d, 3H), 1.09 (d, 3H), 0.87 (t, 3H).
A solution of example 7 (0.500 g) and intermediate 52 (0.180 g) in anhydrous acetonitrile (7 mL) was stirred at room temperature for 2 h under nitrogen atmosphere. The solvent was evaporated and the residue dissolved in anhydrous MeOH (5 mL). Then sodium cyanoborohydride (1M in THF, 0.375 ml) and acetic acid (0.045 ml) were added under nitrogen atmosphere and the mixture stirred overnight. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.170 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 9.00 (d, 1H), 8.45 (d, 1H), 8.11 (d, 1H), 7.59 (d, 1H), 7.39 (d, 1H), 7.29 (m, 1H), 5.55 (dd, 1H), 4.29 (d, 1H), 4.24 (d, 1H), 4.13 (s, 1H), 4.20-4.0 (m, 2H), 3.86 (q, 1H), 3.55 (m, 1H), 3.19 (m, 1H), 3.09 (m, 1H), 3.05 (m, 1H), 2.93 (m, 2H), 2.67 (s, 3H), 2.52 (m, 1H), 2.49 (m, 1H), 2.32 (d, 1H), 2.27 (s, 6H), 2.02 (m, 1H), 1.94 (m, 2H), 1.80 (m, 2H), 1.65 (m, 1H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.33 (s, 3H), 1.31 (d, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.16 (d, 3H), 1.09 (d, 3H), 0.87 (t, 3H).
A solution of example 6 (0.100 g) and intermediate 54 (0.050 g) in anhydrous DCM (3 mL) was stirred at room temperature for 24 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.120 mL) and acetic acid (0.008 mL) were added under nitrogen atmosphere and the mixture stirred for 1 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (5 mL) and extracted with DCM (3×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 95\5 to 90\10) to give a compound that was dissolved in MeOH (2 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.015 g).
1H-NMR (CDCl3) δ: 8.98 (d, 1H), 8.46 (d, 1H), 8.09 (d, 1H), 7.56 (d, 1H), 7.36 (d, 1H), 7.30 (m, 1H), 5.67 (dd, 1H), 4.30 (d, 1H), 4.24 (d, 1H), 4.13 (s, 1H), 4.00 (m, 2H), 3.85 (q, 1H), 3.56 (m, 1H), 3.19 (m, 1H), 3.09 (m, 1H), 3.03 (m, 1H), 2.91 (m, 2H), 2.64 (s, 3H), 2.58 (m, 1H), 2.47 (m, 1H), 2.28 (m+s, 6H +1H), 2.00 (m, 1H), 1.89 (m, 2H), 1.79 (m, 1H), 1.70 (m, 2H), 1.60-1.50 (m, 2H), 1.48 (s, 3H), 1.39 (d, 3H), 1.31 (s, 3H), 1.31 (d, 3H), 1.25 (d, 3H), 1.25 (m, 1H), 1.15 (d, 3H), 1.09 (d, 3H), 0.83 (t, 3H).
A solution of example 6 (0.080 g) and intermediate 56 (0.042 g) in anhydrous DCM (5 mL) was stirred at room temperature for 1 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.120 mL) and HCl (2N in Et2O, 0.010 mL) were added under nitrogen atmosphere and the mixture stirred for 1 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 96\4 to 90\10) to give a compound that was dissolved in MeOH (3 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.013 g).
1H-NMR (CDCl3) δ: 8.99 (d, 1H), 8.47 (d, 1H), 8.10 (d, 1H), 7.55 (s, 1H), 7.31 (s, 1H), 7.29 (m, 1H), 5.67 (dd, 1H), 4.30 (d, 1H), 4.25 (d, 1H), 4.11 (s, 1H), 3.96 (m, 2H), 3.85 (q, 1H), 3.58 (m, 1H), 3.18 (dd, 1H), 3.09 (m, 1H), 3.02 (m, 1H), 2.90-2.75 (m, 2H), 2.67 (s, 3H), 2.58 (m, 1H), 2.46 (m, 1H), 2.29 (d, 1H), 2.27 (s, 6H), 1.90-1.86 (m, 3H), 1.78 (m, 1H), 1.70 (m, 2H), 1.60-1.50 (m, 3H), 1.47 (s, 3H), 1.5-1.4 (m, 4H), 1.38 (d, 3H), 1.33 (s, 3H), 1.32 (d, 3H), 1.28 (d, 3H), 1.25 (m, 1H), 1.15 (d, 3H), 1.09 (d, 3H), 0.85 (t, 3H).
A solution of intermediate 51 (0.040 g) in acetonitrile (1 mL) and a 2M HCl aqueous solution (3 mL) was heated to 80° C. for 3 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 6 (0.100 g) dissolved in anhydrous acetonitrile (4 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 2 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.150 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (5 mL) was added and the product was extracted with DCM (3×10 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH 94\6) affording the title compound (0.027 g).
1H-NMR (CDCl3) δ: 9.04 (d, 1H), 8.46 (dd, 1H), 8.12 (d, 1H), 7.61 (d, 1H), 7.56 (m, 1H), 7.30 (m, 1H), 5.63 (dd, 1H), 4.27 (d, 1H), 4.22 (d, 1H), 4.19 (d, 1H), 4.20-4.08 (m, 2H), 3.84 (q, 1H), 3.54 (m, 1H), 3.36-3.19 (m, 2H), 3.16 (dd, 1H), 3.05 (m, 2H), 2.55 (m, 1H), 2.44 (m, 1H), 2.43 (s, 3H), 2.28 (s, 1H), 2.27 (s, 6H), 1.85 (m, 1H), 1.75 (m, 1H), 1.70-1.65 (m, 2H), 1.55 (m, 1H), 1.48 (s, 3H), 1.40 (d, 3H), 1.30 (d, 3H), 1.24-1.20 (m, 4H), 1.23 (s, 3H), 1.14 (d, 3H), 1.09 (d, 3H), 0.82 (t, 3H).
A solution of intermediate 51 (0.530 g) in acetonitrile (30 mL) and a 3M HCl aqueous solution (20 mL) was heated to 80° C. for 3 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.500 g) dissolved in anhydrous acetonitrile (2 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 1 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.75 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (15 mL) was added and the mixture was extracted with DCM (3×20 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (10 mL) and stirred overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with DCM\MeOH from 98\2 to 90\10) to give the title compound (0.350 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 9.04 (d, 1H), 8.46 (dd, 1H), 8.12 (d, 1H), 7.61 (d, 1H), 7.56 (m, 1H), 7.30 (m, 1H), 5.63 (dd, 1H), 4.27 (d, 1H), 4.22 (d, 1H), 4.19 (d, 1H), 4.20-4.08 (m, 2H), 3.84 (q, 1H), 3.54 (m, 1H), 3.36-3.19 (m, 2H), 3.16 (dd, 1H), 3.05 (m, 2H), 2.55 (m, 1H), 2.44 (m, 1H), 2.43 (s, 3H), 2.28 (s, 1H), 2.27 (s, 6H), 1.85 (m, 1H), 1.75 (m, 1H), 1.70-1.65 (m, 2H), 1.55 (m, 1H), 1.48 (s, 3H), 1.40 (d, 3H), 1.30 (d, 3H), 1.24-1.20 (m, 4H), 1.23 (s, 3H), 1.14 (d, 3H), 1.09 (d, 3H), 0.82 (t, 3H).
A solution of example 6 (0.084 g) and intermediate 63 (0.040 g) in anhydrous DCM (4 mL) was stirred at room temperature for 1 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.125 mL) and acetic acid (0.007 mL) were added and the mixture stirred overnight. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with DCM\MeOH 90\10) to give a compound that was dissolved in MeOH (3 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.006 g).
1H-NMR (CDCl3) δ: 8.81 (d, 1H), 8.11 (d, 1H), 8.06 (d, 1H), 7.70 (t, 1H), 7.57 (t, 1H), 7.26 (d, 1H), 5.71 (dd, 1H), 4.31 (d, 1H), 4.26 (d, 1H), 4.12 (d, 1H), 3.85 (q, 1H), 3.55 (m, 1H), 3.19 (m, 1H), 3.10 (m, 1H), 3.07 (m, 2H), 3.03 (m, 1H), 2.90 (m, 1H), 2.80 (m, 1H), 2.68 (s, 3H), 2.58 (m, 1H), 2.45 (m, 1H), 2.30 (d, 1H), 2.27 (s, 6H), 1.90 (m, 1H), 1.75 (m, 2H), 1.70 (m, 3H), 1.52 (m, 2H), 1.48 (s, 3H), 1.37 (d, 3H), 1.33 (s, 3H), 1.32 (d, 3H), 1.24 (m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.09 (d, 3H), 0.85 (t, 3H).
A solution of example 6 (0.100 g) and 3-quinolin-4-yl-propionaldehyde (0.031 g) in anhydrous DCM (4 mL) was stirred at room temperature for 2 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.150 mL) and acetic acid (0.009 mL) were added and the mixture stirred overnight. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with DCM\MeOH 95\5) to give a compound that was dissolved in MeOH (3 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.011 g).
1H-NMR (CDCl3) δ: 8.80 (d, 1H), 8.13-8.09 (d, 2H), 7.70 (t, 1H), 7.58 (t, 1H), 7.30 (m, 1H), 5.80 (dd, 1H), 4.31 (d, 1H), 4.26 (d, 1H), 4.19 (s, 1H), 3.86 (q, 1H), 3.56 (m, 1H), 3.50 (bm, 1H), 3.26 (m, 1H), 3.19 (m, 1H), 3.12 (m, 1H), 3.06 (m, 3H), 2.90 (m, 1H), 2.69 (s, 3H), 2.58 (m, 1H), 2.46 (m, 1H), 2.36 (d, 1H), 2.27 (s, 6H), 1.92 (m, 3H), 1.80-1.70 (m, 2H), 1.65 (m, 1H), 1.60 (m, 1H), 1.50 (s, 3H), 1.40 (d, 3H), 1.33 (s+d, 3H+3H), 1.24 (m, 4H), 1.16 (d, 3H), 1.11 (d, 3H), 0.87 (t, 3H).
A solution of example 6 (0.130 g) and intermediate 59 (0.047 g) in anhydrous THF (5 mL) was stirred at room temperature for 1 hr under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.195 mL) and acetic acid until pH=5 were added and the mixture stirred for 3 h. The reaction was quenched with a saturated NaHCO3 aqueous solution (10 mL) and extracted with DCM (3×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 98\2 to 96\4) to give a compound that was dissolved in MeOH (5 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.015 g).
m\z ([MH]+)=810.
A solution of intermediate 57 (0.270 g) in acetonitrile (4 mL) and 3M HCl aqueous solution (4 mL) was heated to 50° C. for 24 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 6 (0.500 g) in anhydrous acetonitrile (4 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 3 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.75 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (15 mL) was added and the mixture was extracted with DCM (3×20 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The residue was dissolved in MeOH (10 mL) and stirred overnight. After solvent evaporation the crude material was purified by flash chromatography (eluting with DCM\MeOH from 98\2 to 90\10) to give the title compound (0.190 g).
1H-NMR (CDCl3) δ: 8.79 (d, 1H), 8.09 (m, 1H), 7.69 (t, 1H), 7.56 (t, 1H), 7.38 (d, 1H), 5.68 (dd, 1H), 4.29 (d, 1H), 4.22 (d, 1H), 4.21 (s, 1H), 3.83 (q, 1H), 3.54 (m, 1H), 3.41-3.1 (m, 4H), 3.17 (dd, 1H), 3.07 (m, 1H), 3.02 (m, 1H), 2.45 (m, 2H +—OCH3), 2.30 (m, 1H), 2.27 (s, N(CH3)2), 1.88 (m, 1H), 1.75 (m, 2H), 1.66 (m, 1H), 1.55 (m, 1H), 1.47 (s, 3H), 1.37 (d, 3H), 1.30 (d, 3H), 1.25 (d, 3H), 1.25 (m, 4H), 1.12 (d, 3H), 1.08 (d, 3H), 0.85 (t, 3H).
A solution of intermediate 57 (0.832 g) in acetonitrile (16 mL) and a 3M HCl aqueous solution (16 mL) was heated to 50° C. for 16 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (1.500 g) in anhydrous acetonitrile (12 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 1 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 3.3 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent water (100 mL) was added and the mixture extracted with EtOAc (2×100 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (DCM\MeOH 95\5). The obtained compound was dissolved in MeOH (20 mL) and stirred overnight. After solvent evaporation the compound was purified by flash chromatography (eluting with DCM\MeOH 95\5) to give the title compound (0.365 g, (21S) isomer 95% pure by NMR analysis). 1H-NMR (CDCl3) 6: 8.79 (d, 1H), 8.09 (m, 1H), 7.69 (t, 1H), 7.56 (t, 1H), 7.38 (d, 1H), 5.68 (dd, 1H), 4.29 (d, 1H), 4.22 (d, 1H), 4.21 (s, 1H), 3.83 (q, 1H), 3.54 (m, 1H), 3.41-3.1 (m, 4H), 3.17 (dd, 1H), 3.07 (m, 1H), 3.02 (m, 1H), 2.45 (m, 2H +3H), 2.30 (m, 1H), 2.27 (s, 6H), 1.88 (m, 1H), 1.75 (m, 2H), 1.66 (m, 1H), 1.55 (m, 1H), 1.47 (s, 3H), 1.37 (d, 3H), 1.30 (d, 3H), 1.25 (d, 3H), 1.25 (m, 4H), 1.12 (d, 3H), 1.08 (d, 3H), 0.85 (t, 3H).
A solution of example 7 (0.050 g) and quinoline-4-carbaldehyde (0.020 g) in anhydrous toluene (3 mL) was stirred at 100° C. for 16 h under nitrogen atmosphere. After evaporation of the solvent the crude material was dissolved in anhydrous MeOH (5 mL) palladium (10 wt. % on carbon powder, 0.006 g) was added and the mixture stirred under hydrogen atmosphere (1 atm) for 5 h. Filtration through a celite pad eluting with MeOH and purification by flash chromatography (eluting with: DCM\MeOH 93\7) gave the title compound (0.007 g).
m\z ([MH]+)=768
1H-NMR (CDCl3) δ: 8.89 (d, 1H), 8.11 (d, 1H), 8.09 (d, 1H), 7.69 (t, 1H), 7.60 (t, 1H), 7.56 (t, 1H), 4.65 (m, 1H), 4.48 (m, 1H), 4.38 (m, 1H), 3.09 (m, 1H), 2.43 (m, 1H), 1.14 (d, 3H).
A solution of 3-(5-methyl-2-furyl)butanal (0.030 g) and example 6 (0.067 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 5 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.250 mL) and acetic acid (0.025 mL) were added and the mixture was stirred at room temperature for 24 h. MeOH (1 mL) was added and the reaction mixture heated to 60° C. for 24 h. After evaporation under reduced pressure the residue was dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH 95\5\0.5) to give the title compound (0.029 g).
1H-NMR (CDCl3) δ: 5.82 (m, 2H), 4.10 (m, 1H), 3.02 (m, 1H), 2.86 (s, 2H), 2.77 (m, 1H), 2.40-2.20 (m, 3H+1H), 1.80-1.60 (m, 2H), 1.08 (d, 3H).
A solution of 3-(5-methyl-2-furyl)butanal oxalate (0.045 g), example 6 (0.067 g) and DIPEA (0.051 mL) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 1.5 h under nitrogen atmosphere. Then sodium cyanoborohydride (1M in THF, 0.250 mL) and acetic acid (0.025 mL) were added and the mixture was stirred at room temperature for 15 h. MeOH (1 mL) was added and the reaction mixture heated to 60° C. for 48 h. After evaporation under reduced pressure the residue was dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM, DCM\MeOH\NH4OH from 96\4\0.1 to 92\8\0.2) to give the title compound (0.016 g).
m\z ([MH]+)=744.
1H-NMR (CDCl3) δ: 8.52 (d, 2H), 7.28 (d, 2H), 6.59 (dd, 2H), 4.24 (m, 1H), 3.65 (m, 2H), 3.05 (m, 1H), 2.38 (m, 1H), 1.10 (d, 3H).
To a solution of example 7 (0.030 g) in anhydrous THF (1 mL) intermediate 75 (0.050 g) was added and the mixture was stirred at room temperature for 3 h under nitrogen atmosphere. Then sodium cyanoborohydride (1 M in THF, 0.150 mL) and acetic acid (0.010 mL) were added and the mixture was allowed to react for 4 h. The solvent was evaporated under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (1 mL) and the mixture stirred overnight. After evaporation of the solvent the crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound ((0.014 g).
1H-NMR (CDCl3) δ: 7.83 (s, 1H), 7.73 (s, 1H), 7.03 (dd, 1H), 6.62 (tt, 1H), 4.28 (m, 2H), 4.12 (s, 1H), 3.04 (m, 1H), 2.91 (m, 1H), 2.82 (m, 1H), 2.32 (bs, 2H), 2.07 (m, 1H), 2.00 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 76 (0.025 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 5 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the residue dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.005 mL) were added. After 18 h the solvent was evaporated under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.028 g).
1H-NMR (CDCl3) δ: 7.55 (d, 2H), 7.33 (d, 2H), 4.15 (m, 1H), 4.00 (m, 1H), 3.85 (m, 1H), 3.05 (m, 1H), 3.02-2.90 (m, 2H), 2.42 (s, 3H), 2.35 (s, 3H), 2.33 (m, 1H), 1.90 (m, 1H), 1.72 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 77 (0.023 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.005 mL) were added. After 24 h the solvent was evaporated under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 8.24 (d, 2H), 7.96 (d, 2H), 7.61 (d, 1H), 7.54 (d, 1H), 4.14 (m, 1H), 4.20÷4.06 (m, 2H), 3.00÷2.88 (m, 2H), 2.32 (m, 1H), 2.03 (m, 1H), 1.92 (m, 1H).
A solution of example 7 (0.050 g) and intermediate 78 (0.025 g) in anhydrous acetonitrile (2 mL) was stirred at room temperature for 16 h under nitrogen atmosphere then heated to 50° C. for 4 h. After evaporating the solvent under reduced pressure, the crude was dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.100 mL) and acetic acid (0.006 mL) were added. After 24 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 8.57 (d, 1H), 7.70 (d, 1H), 7.62 (s, 1H), 7.52 (s, 1H), 4.20-4.16 (m, 2H), 3.05 (m, 1H), 2.96-2.88 (m, 2H), 2.32 (m, 1H), 2.02 -1.92 (m+m, 2H), 1.10 (d, 3H).
A solution of example 7 (0.020 g) and intermediate 79 (0.015 g) in anhydrous acetonitrile (2 mL) was stirred at room temperature for 3 h under nitrogen atmosphere then heated to 40° C. for 4 h. After evaporating the solvent under reduced pressure, the crude material was dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.015 mL) and acetic acid (0.003 mL) were added. After 24 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 1 00\0 to 98\2) to give the title compound (0.004 g).
1H-NMR (CDCl3) δ: 7.53 (s, 1H), 4.12 (d, 1H), 3.03 (m, 1H), 2.92 (m, 1H), 2.84 (m, 1H), 2.70-2.60 (m, 2H), 2.11 (m, 1H), 1.70 (m, 2H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 80 (0.038 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature overnight under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 24 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 98\2\0 to 95\5\0.5) and by preparative TLC (DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.017 g).
1H-NMR (CDCl3) δ: 7,78 (d, 2H), 7.64 (d, 2H), 7.59 (s, 1H), 5.60 (dd, 1H), 4.30 (d, 1H), 4.25 (d, 1H), 4.15 (s, 1H), 4.10 (m, 1H), 3.96 (m, 1H), 3.86 (q, 1H), 3.56 (m, 1H), 3.19 (dd, 1H), 3.09 (m, 1H), 3.04 (m, 1H), 2.96 (m, 2H), 2.68 (s, 3H), 2.58 (m, 1H), 2.45 (m, 1H), 2.42 (s, 3H), 2.33 (s, 1H), 2.27 (s, 6H), 2.00÷1.88 (2H), 1.90 (m, 1H), 1.81 (dd, 1H), 1.73 (bd, 1H), 1.67 (m, 1H), 1.60 (m, 1H), 1.49 (s, 3H), 1.38 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.25 (d, 3H), 1.16 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 81 (0.020 g) in anhydrous acetonitrile (0.7 mL) was stirred at room temperature for 3 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1M in THF, 0.035 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.009 g).
m\z ([MH]+)=746.
1H-NMR (CDCl3) δ: 8.47-8.40 (m, 2H), 7.51 (m, 1H), 7.19 (m, 1H), 4.14 (m, 1H), 3.03 (m, 1H), 2.98-2.85 (m, 2H), 2.75-2.60 (m, 2H), 2.31 (bm, 1H), 1.82-1.76 (m, 2H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 82 (0.025 g) in anhydrous acetonitrile (3 mL) was stirred at 50° C. for 3 h and at room temperature overnight under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2.2 mL) then sodium cyanoborohydride (1M in THF, 0.040 mL) and acetic acid (0.020 mL) were added. After 10 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.026 g).
1H-NMR (CDCl3) δ: 8.54 (d, 1H), 8.01 (s, 1H), 7.98 (s, 1H), 7.65 (t, 1H), 7.50 (t, 1H), 7.07 (m, 1H), 4.27 (d, 1H), 4.14 (m, 2H), 3.04 (m, 1H), 2.89 (m, 2H), 2.34 (m, 1H), 2.12 (m, 1H), 2.03 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 83 (0.050 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature overnight under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 24 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 90\10) and by preparative TLC (eluting with: DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 8.47 (d, 2H), 7.13 (d, 2H), 5.70 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.14 (s, 1H), 3.85 (q, 1H), 3.56 (m, 1H), 3.18 (dd, 1H), 3.10 (m, 1H), 3.03 (q, 1H), 2.95÷2.83 (m, 2H), 2.65÷2.55 (m, 2H), 2.67 (s, 3H), 2.59 (m, 1H), 2.46 (m, 1H), 2.31 (s, 1H), 2.27 (s, 6H), 1.91 (m, 1H), 1.82-1.76 (m, 3H), 1.73 (m, 1H), 1.68 (m, 1H), 1.58 (m, 1H), 1.48 (s, 3H), 1.39 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.09 (d, 3H), 0.87 (t, 3H).
A solution of example 7 (0.028 g) and intermediate 84 (0.018 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 30 min then heated to 50° C. for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.025 mL) and acetic acid (0.004 mL) were added. After 24 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 1 00\0 to 97\3) to give the title compound (0.005 g).
1H-NMR (CDCl3) δ: 9.09 (d, 1H), 8.63 (d, 1H), 8.18 (s, 1H), 8.07 (s, 1H), 7.47 (d, 1H), 4.30 (m, 2H), 4.13 (s, 1H), 3.04 (m, 1H), 2.92-2.80 (m, 2H), 2.33 (m, 1H), 2.11 (m, 1H), 2.00 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 85 (0.018 g) in anhydrous acetonitrile (2.2 mL) was stirred at 50° C. for 16 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (3 mL) then sodium cyanoborohydride (1M in THF, 0.040 mL) and acetic acid (0.020 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a NaHCO3 saturated aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 95\5) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 8.55 (d, 1H), 7.96 (s, 1H), 7.85 (s, 1H), 7.42 (d, 1H), 4.13 (d, 1H), 3.04 (m, 1H), 2.89 (m, 1H), 2.84 (m, 1H), 2.33 (m, 1H), 2.10 (m, 1H), 1.95 (m, 1H), 1.10 (d, 3H).
A solution of example 7 (0.030 g) and intermediate 86 (0.029 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 5 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.022 mL) and acetic acid (0.003 mL) were added. After 12 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.022 g).
1H-NMR (CDCl3) δ: 7.47 (d, 2H), 7.15 (t, 1H), 4.11 (m, 1H), 3.98 (m, 1H), 3.82 (m, 1H), 2.93 (m, 2H), 2.38 (s, 3H), 2.33 (s, 3H), 2.29 (m, 1H), 1.6÷1.8 (m, 2H).
A solution of example 7 (0.040 g) and intermediate 87 (0.042 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 4 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.030 mL) and acetic acid (0.004 mL) were added. After 12 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.026 g).
1H-NMR (CDCl3) 6: 7.84 (bs, 1H), 7.74 (m, 1H), 7.43 (m, 2H), 4.12 (s, 1H), 3.99 (m, 1H), 3.82 (m, 1H), 2.9÷2.88 (m, 2H), 2.40 (s, 3H), 2.33 (s, 3H), 2.30 (s, 1H), 1.85÷1.75 (m, 2H).
A solution of example 7 (0.037 g) and intermediate 88 (0.022 g) in anhydrous acetonitrile (1.5 mL) was stirred at 50° C. for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.030 mL) and acetic acid (0.004 mL) were added. After 18h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 8.17 (s, 1H), 8.14 (s, 1H), 7.68 (dd, 1H), 7.51 (dd, 1H), 7.29 (m, 2H), 4.34-4.29 (m, 2H), 4.14 (s, 1H), 3.03 (m, 1H), 2.90 (m, 2H), 2.33 (bm, 1H), 2.15 (m, 1H), 2.00 (m, 1H), 1.09 (d, 3H)
A solution of example 7 (0.050 g) and intermediate 89 (0.036 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 20 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.075 mL) and acetic acid (0.010 mL) were added. After 20 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.011 g).
1H-NMR (CDCl3) δ: 8.60 (d, 1H), 7.69 (d, 1H), 7.52 (d, 1H), 6.62 (d, 1H), 4.29 (m, 2H), 4.14 (m, 1H), 3.04 (m, 1H), 2.88 (m, 2H), 2.33 (m, 1H), 2.20 (m, 1H), 1.98 (m, 1H), 1.10 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 90 (0.025 g) in anhydrous acetonitrile (2 mL) was stirred at 50° C. for 6 h then at room temperature overnight under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.150 mL) and acetic acid (0.006 mL) were added. After 72 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 90\10) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 8.27 (d, 1H), 7.59 (d, 1H), 7.19 (m, 1H), 7.16 (m, 1H), 4.30-4.10 (m, 3H), 3.15 (m, 2H), 2.96 (m, 1H), 2.31 (m, 1H), 2.00 -1.82 (m, 2H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 91 (0.032 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2 to 94\6) and by preparative TLC (DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.018 g). 1H-NMR (CDCl3) 6: 8.20 (s, 1H), 8.14 (s, 1H), 8.12 (d, 1H), 8.04 (d, 1H), 7.76 (d, 1H), 7.67 (t, 1H), 7.66 (d, 1H), 7.45 (t, 1H), 5.68 (dd, 1H), 4. 31 (m, 3H), 4.26 (d, 1H), 4.15 (s, 1H), 3.86 (q, 1H), 3.56 (m, 1H), 3.18 (dd, 1H), 3.11 (m, 1H), 3.05 (m, 1H), 2.95÷2.80 (m, 2H), 2.70 (s, 3H), 2. 58 (m, 1H), 2.46 (m, 1H), 2.35 (s, 1H), 2.27 (s, 6H), 2.14 (m, 1H), 2.06 (m, 1H), 1.96 (m, 1H), 1.80 (m, 1H), 1.74 (m, 1H), 1.70 (m, 1H), 1.65 (m, 1H), 1.50 (s, 3H), 1.40 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.16 (d, 3H), 1.10 (d, 3H), 0.90 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 92 (0.034 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1 M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2 to 94\6) to give the title compound (0.020 g).
1H-NMR (CDCl3) δ: 8.89 (d, 1H), 8.25 (d, 1H), 8.14 (d, 1H), 7.87 (s, 1H), 7.84 (s, 1H), 7.73 (m, 1H), 7.59 (m, 1H), 7.42 (d, 1H), 5.63 (dd, 1H), 4.37 (m, 2H), 4.30 (d, 1H), 4.24 (d, 1H), 4.16 (s, 1H), 3.85 (q, 1H), 3.55 (m, 1H), 3.18 (dd, 1H), 3.10 (m, 1H), 3.04 (m, 1H), 2.94 (m, 2H), 2.70 (s, 3H), 2.58 (m, 1H), 2.45 (m, 1H), 2.34 (s, 1H), 2.27 (s, 6H), 2.19 (m, 1H), 2.05 (m, 1H), 1.88 (m, 1H), 1.80 (dd, 1H), 1.73 (m, 1H), 1.67 (m, 1H), 1.56 (m, 1H), 1.49 (s, 3H), 1.38 (d, 3H), 1.33 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.24 (d, 3H), 1.15 (d, 3H), 1.10 (d, 3H), 0.83 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 93 (0.030 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2 to 94\6) and by preparative TLC (DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.006 g).
1H-NMR (CDCl3) δ: 9.10 (s, 1H), 8.28 (s, 1H), 8.22 (s, 1H), 8.03 (m, 2H), 7.73 (t, 1H), 7.66 (t, 1H), 5.63 (dd, 1H), 4.35 (m, 2H), 4.30 (d, 1H), 4.26 (d, 1H), 4.15 (m, 1H), 3.87 (q, 1H), 3.55 (m, 1H), 3.17 (m, 1H), 3.11 (m, 1H), 3.05 (m, 1H), 3.00-2.80 (m, 2H), 2.71 (s, 3H), 2.60 (m, 1H), 2.46 (m, 1H), 2.35 (m, 1H), 2.27 (s, 6H), 2.15 (m, 1H), 2.06 (m, 1H), 1.96 (m, 1H), 1.80 (m, 1H), 1.74 (m, 1H), 1.68 (m, 1H), 1.60 (m, 1H), 1.50 (s, 3H), 1.40 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.30 (m, 1H), 1.24 (d, 3H), 1.16 (d, 3H), 1.10 (d, 3H), 0.90 (t, 3H).
A solution of example 7 (0.042 g) and intermediate 94 (0.025 g) in anhydrous acetonitrile (2 mL) was stirred at 50° C. for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.035 mL) and acetic acid (0.004 mL) were added. After 18 h at room temperature the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 98\2) to give the title compound (0.012 g).
1H-NMR (CDCl3) δ: 7.21 (m, 1H), 6.94 (m, 2H), 4.13 (m, 1H), 3.03 (m, 1H), 2.91 (m, 1H), 2.83 (m, 1H), 2.80-2.60 (m, 2H), 2.31 (m, 1H), 1.80 (m, 2H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 95 (0.037 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.075 mL) and acetic acid (0.009 mL) were added. After 20 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give the title compound (0.01 8 g).
1H-NMR (CDCl3) δ: 8.44 (d, 1H), 8.36 (d, 1H), 7.54 (d, 1H), 6.82 (d, 1H), 4.40-4.20 (m, 2H), 4.14 (m, 1H), 3.04 (m, 1H), 2.90 (s, 3H), 2.89 (m, 2H), 2.33 (m, 1H), 2.15 (m, 1H), 2.05 (m, 1H), 1.10 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 96 (0.037 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 3 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.075 mL) and acetic acid (0.009 mL) were added. After 20 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2) to give the title compound (0.020 g).
1H-NMR (CDCl3) δ: 7.66 (m, 1H), 7.40 (m, 1H), 7.35 (m, 1H), 7.20 (m, 1H), 4.15 (m, 1H), 4.25 (m, 1H), 4.13 (m, 1H), 3.05 (m, 1H), 2.95 (m, 2H), 2.79 (m, 2H), 2.36 (m, 1H), 2.00 (m, 1H), 1.86 (m, 1H), 1.10 (d, 3H).
A solution of example 7 (0.040 g) and intermediate 97 (0.035 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 12 h then heated to 50° C. for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.030 mL) and acetic acid (0.004 mL) were added. After 12 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.017 g). 1H-NMR (CDCl3) 6: 7.73 (d, 2H), 7.35 (d, 2H), 6.35 (s, 1H), 4.15 (m, 1H), 2.93 (m, 2H), 2.82 (m, 1H), 2.34 (m, 1H), 1.88 (m, 1H).
A solution of example 7 (0.040 g) and intermediate 98 (0.042 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 3 h then heated to 50° C. for 3 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.060 mL) and acetic acid (0.008 mL) were added. After 6 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 98\2) to give the title compound (0.005 g). 1H-NMR (CDCl3) 6: 8.85 (m, 1H), 8.25 (s, 1H), 4.63 (m, 1H), 4.45 (m, 1H), 4.15 (m, 1H), 2.97 (m, 2H), 2.76 (s, 3H), 2.33 (m, 1H), 2.0÷2.2 (m, 2H).
A solution of example 7 (0.050 g) and intermediate 99 (0.031 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature overnight under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.074 mL) and acetic acid (0.010 mL) were added. After 4 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.015 g).
1H-NMR (CDCl3) δ: 8.54 (s, 1H), 8.07 (s, 1H), 5.58 (dd, 1H), 4.37 (t, 2H), 4.31 (d, 1H), 4.25 (d, 1H), 4.19 (s, 3H), 4.13 (bs, 1H), 3.86 (q, 1H), 3.55 (m, 1H), 3.20 (m, 1H), 3.10 (m, 1H), 3.04 (m, 1H), 2.89 (bm, 2H), 2.70 (s, 3H), 2.59 (m, 1H), 2.48 (bm, 1H), 2.34 (s, 1H), 2.29 (s, 6H), 2.12 (m, 1H), 2.04 (m, 1H), 1.95 (m, 1H), 1.85-1.48 (m, 5H), 1.49 (s, 3H), 1.39 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (d+m, 3H+1H), 1.16 (d, 3H), 1.10 (d, 3H), 0.89 (t, 3H).
A solution of example 7 (0.043 g) and intermediate 100 (0.025 g) in anhydrous acetonitrile (2 mL) was stirred at room temperature for 30 min then heated to 50° C. for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.030 mL) and acetic acid (0.005 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 7.05 (d, 1H), 6.82 (d, 1H), 6.47 (dd, 1H), 4.13 (s, 1H), 3.87 (m, 2H), 3.81 (s, 3H), 3.04 (m, 1H), 2.97-2.92 (m, 2H), 2.34 (m, 1H), 1.96 (m, 1H), 1.80 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 101 (0.042 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (1.5 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.005 mL) were added. After 8 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH 95\5\0.5) to give the title compound (0.006 g).
1H-NMR (CDCl3) δ: 8.30 (dd, 1H), 7.87 (dd, 1H), 7.30 (d, 1H), 7.03 (dd, 1H), 6.43 (d, 1H), 5.71 (dd, 1H), 4.44÷4.36 (m, 2H), 4.32 (d, 1H), 4.25 (d, 1H), 4.13 (s, 1H), 3.85 (q, 1H), 3.55 (m, 1H), 3.22 (dd, 1H), 3.11 (m, 1H), 3.04 (m, 1H), 2.90÷2.78 (m, 2H), 2.70 (s, 3H), 2.62÷2.50 (m, 2H), 2.35 (s, 1H), 2.32 (s, 6H), 2.06 (m, 1H), 2.00 (m, 1H), 1.95 (m, 1H), 1.80÷1.68 (m, 3H), 1.58 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.33 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.10 (d, 3H), 0.89 (t, 3H).
To a solution of example 13 (0.232 g) in anhydrous DMF (5 mL) 2-[(1,3-thiazol-2-ylamino)carbonyl]benzoic acid (0.097 g), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (0.186 g) and DIPEA (0.120 mL) were added and the resulting mixture was stirred overnight at room temperature. It was diluted with DCM (10 mL) and washed with a 5% NaHCO3 aqueous solution (2×5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in DCM (5 mL) loaded on SCX-cartridge (5 g, loading 0.75 mmol\g, previously washed with 50 mL of MeOH), washed with MeOH (50 mL), then the product eluted with NH3 (0.25M solution in MeOH, 60 mL), followed by MeOH (10 mL). Solvent evaporation gave the title compound (0.170 g).
1H-NMR (CDCl3) δ: 7.80 (m, 2H), 7.67 (m, 2H), 5.53 (dd, 1H), 4.30 (d, 1H), 4.24 (d, 1H), 4.17 (s, 1H), 3.84 (m, 2H), 3.74 (m, 1H), 3.55 (m, 1H), 3.20-3.15 (m, 2H), 3.12-2.88 (m, 3H), 2.63 (s, 3H), 2.54 (m, 1H), 2.47 (m, 1H), 2.30 (m, 1H), 2.27 (s, 6H), 1.86 (m, 1H), 1.75-1.65 (m, 3H), 1.48 (m, 1H), 1.43 (s, 3H), 1.29 (s, 3H), 1.28-1.24 (m, 2*3H+1H), 1.119 (d, 3H), 1.12 (d, 3H), 1.06 (d, 3H), 0.73 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 102 (0.042 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 6 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.070 mL) and acetic acid (0.009 mL) were added. After 20 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 98\2) to give the title compound (0.016 g).
1H-NMR (CDCl3) δ: 7.49 (d, 1H), 6.36 (d, 1H), 5.62 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.23 (m, 2H), 4.13 (s, 1H), 3.85 (q, 1H), 3.56 (m, 1H), 3.19 (m, 1H), 3.10 (m, 1H), 3.03 (m, 1H), 2.92÷2.82 (m, 2H), 2.69 (s, 3H), 2.66 (s, 3H), 2.58 (m, 1H), 2.55 (s, 2.47 (m, 1H), 2.33 (s, 1H), 2.28 (s, 6H), 2.08 (m, 1H), 1.98 (m, 1H), 1.94 (m, 1H), 1.80 (m, 1H), 1.76÷1.66 (m, 2H), 1.62÷1.50 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
and
A solution of example 7 (0.050 g) and intermediate 103 (0.034 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by preparative TLC (DCM\MeOH\NH4OH 90\9\0.5) to give the title compound 108 (0.008 g) and the title compound 109 (0.004 g).
1H-NMR (CDCl3) δ (example 108): 9.11 (s, 1H), 8.44 (d, 1H), 8.08 (s, 1H), 7.47 (d, 1H), 4.41 (m, 1H), 4.28 (m, 1H), 4.14 (m, 1H), 3.05 (m, 1H), 3.00-2.90 (m, 2H), 2.31 (m, 1H), 2.08 (m, 1H), 1.95 (m, 1H), 1.09 (d, 3H).
1H-NMR (CDCl3) δ (example 109): 8.90 (s, 1H), 8.45 (d, 1H), 8.16 (s, 1H), 7.70 (d, 1H), 4.48 (m, 1H), 4.37 (m, 1H), 4.15 (bs, 1H), 3.08 (m, 1H), 2.95 (m, 1H), 2.33 (m, 1H), 2.14 (m, 1H), 2.04 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 104 (0.025 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give the title compound (0.021 g).
1H-NMR (CDCl3) δ: 8.00 (s, 1H), 7.80 (d, 1H), 7.48 (d, 1H), 7.40-7.20 (m, 2H), 4.38 (m, 1H), 4.24 (m, 1H), 4.10 (m, 1H), 3.05 (m, 1H), 3.03 (m, 1H), 2.98 (m, 1H), 2.32 (m, 1H), 2.02 (m, 1H), 1.93 (m, 1H), 1.09 (d, 3H).
A solution of example 7 (0.050 g) and intermediate 105 (0.034 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) and by preparative TLC (eluting with: DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.004 g).
1H-NMR (CDCl3) δ: 8.39 (dd, 1H), 8.21 (s, 1H), 8.05 (dd, 1H), 7.22 (dd, 1H), 4.43 (t, 2H), 4.15 (m, 1H), 3.04 (m, 1H), 2.32 (m, 1H), 1.10 (d, 3H).
A solution of intermediate 106 (0.080 g) in acetonitrile (3 mL) and a 2N HCl aqueous solution (2 mL) was heated to 40° C. for 1 h. The reaction mixture was cooled down to room temperature, then it was added dropwise to a solution of example 7 (0.090 g) dissolved in anhydrous acetonitrile (3 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The solution was stirred at room temperature overnight. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.200 mL). The reaction mixture was stirred for 5 h at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (10 mL) was added and the product was extracted with DCM (4×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The residue was dissolved in MeOH (4 mL) and stirred overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with DCM\MeOH from 98\2 to 96\4) affording the title compound (0.050 g).
m\z ([MH]+)=829.
1H-NMR (CDCl3) δ: 8.57 (s, 1H), 7.99 (d, 1H), 7.96 (d, 1H), 7.69 (t, 1H), 7.60 (t, 1H), 5.71 (dd, 1H), 4.32 (d, 1H), 4.25 (m, 1H), 4.17 (m, 1H), 3.84 (q, 1H), 3.57 (m, 1H), 3.25 (m, 1H), 3.50-2.90 (m, 4H), 3.20-2.90 (m, 4H), 2.69 (s, 3H), 2.58 (m, 1H), 2.37 (s, 6H), 2.34 (m, 1H), 2.10-1.85 (m, 3H), 1.80-1.40 (m, 4H), 1.49 (s, 3H), 1.37 (d, 3H), 1.31 (d, 3H), 1.31 (s, 3H), 1.28 (m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.10 (d, 3H), 0.85 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 107 (0.027 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by preparative TLC (eluting with: DCM\MeOH\NH4OH 90\9\0.5) to give the title compound (0.012 g).
1H-NMR (CDCl3) δ: 7.79 (d, 2H), 7.56 (m, 1H), 7.36 (m, 2H), 7.29 (d, 1H), 7.25 (m, 1H), 4.32 (m, 1H), 4.15 (m, 1H), 4.04 (m, 1H), 3.05 (m, 1H), 3.05-2.95 (m, 2H), 2.33 (m, 1H), 1.98 (m, 1H), 1.80 (m, 1H), 1.09 (d, 3H).
A solution of intermediate 108 (0.060 g) in acetonitrile (1.5 mL) and a 3M HCl aqueous solution (1.5 mL) was stirred at 70° C. for 14 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.030 g) dissolved in anhydrous acetonitrile (1 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 4 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1 M in THF, 0.150 mL). The reaction mixture was stirred for 72 h at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 100\0 to 97\3) and by preparative TLC (eluting with: DCM\MeOH 90\10) to give the title compound (0.004 g).
1H-NMR (CDCl3) δ: 8.57 (d, 2H), 7.72 (d, 2H), 7.71 (m, 1H), 7.60 (m, 1H), 5.63 (dd, 1H), 4.27 (d, 1H), 4.19 (s, 1H), 4.18 (d, 1H), 4.12 (m, 2H), 3.85 (q, 1H), 3.55 (m, 1H), 3.32 (m, 1H), 3.24 (m, 1H), 3.16 (dd, 1H), 3.05 (m, 1H), 3.03 (m, 1H), 2.56 (m, 1H), 2.44 (m, 1H), 2.41 (s, 3H), 2.27 (s, 1H), 2.26 (s, 6H), 1.85 (m, 1H), 1.78 (m, 1H), 1.70÷1.63 (m, 2H), 1.55 (m, 1H), 1.48 (s, 3H), 1.41 (d, 3H), 1.31 (d, 3H), 1.24 (m, 1H), 1.24 (d, 3H), 1.23 (s, 3H), 1.14 (d, 3H), 1.09 (d, 3H), 0.81 (t, 3H).
A solution of intermediate 109 (0.067 g) in acetonitrile (3 mL) and a 2N HCl aqueous solution (2 mL) was stirred overnight. Then, the reaction mixture was added dropwise to a solution of example 7 (0.090 g) dissolved in anhydrous acetonitrile (3 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The reaction mixture was stirred at room temperature for 3 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.200 mL). The reaction mixture was stirred for 5 h at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (10 mL) was added and the mixture was extracted with DCM (4×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The residue was dissolved in MeOH (4 mL) and stirred overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with DCM\MeOH from 98\2 to 96\4) to give the title compound (0.076 g).
m\z ([MH]+)=815.
1H-NMR (CDCl3) δ: 8.55 (s, 1H), 7.99 (d, 1H), 7.93 (d, 1H), 7.68 (t, 1H), 7.60 (t, 1H), 5.68 (dd, 1H), 4.32 (d, 1H), 4.26 (d, 1H), 4.21 (m, 1H), 3.83 (q, 1H), 3.62-3.48 (m, 3H), 3.23 (m, 2H), 3.19 (dd, 1H), 3.10 (m, 1H), 3.04 (m, 1H), 2.74 (s, 3H), 2.58 (m, 1H), 2.48 (m, 1H), 2.35 (s, 1H), 2.29 (s, 6H), 1.91 (m, 1H), 1.80-1.67 (m, 3H), 1.55 (m, 1H), 1.49 (s, 3H), 1.35 (d, 3H), 1.30 (d, 3H), 1.34 (s, 3H), 1.26 (m, 1H), 1.25 (d, 3H), 1.14 (d, 3H), 1.09 (d, 3H), 0.86 (t, 3H).
A solution of example 6 (0.040 g) and intermediate 110 (0.033 g) in anhydrous acetonitrile (1.5 mL) was stirred at room temperature for 12 h. After evaporating the solvent the residue was dissolved in anhydrous MeOH (1.5 mL) and sodium cyanoborohydride (1M in THF, 0.030 mL) and acetic acid (0.004 mL) were added. The reaction mixture was stirred for 12 h. The solvent was evaporated under vacuum, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×2 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 100\0, 98\2, 97\3) affording the title compound (0.025 g).
1H-NMR (CDCl3) δ: 7.52 (d, 1H), 7.18 (d, 1H), 7.27 (m, 1H), 7.16 (d, 1H), 7.02 (dd, 1H), 5.58 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.14 (m, 1H), 4.12 (m, 1H), 4.02 (m, 1H), 3.86 (q, 1H), 3.56 (m, 1H), 3.20 (m, 1H), 3.10 (m, 1H), 3.04 (m, 1H), 2.93 (m, 2H), 2.68 (s, 3H), 2.60 (m, 1H), 2.50 (m, 1H), 2.32 (m, 1H), 2.30 (s, 6H), 2.05-1.85 (m, 3H), 1.80 (m, 1H), 1.70 (m, 2H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (d+m, 3H+1H), 1.16 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.750 g) and intermediate 110 (0.300 g) in anhydrous acetonitrile (15 mL) was stirred at room temperature for 12 h. After evaporating the solvent the residue was dissolved in anhydrous MeOH (15 mL) and sodium cyanoborohydride (1M in THF, 0.728 mL) and acetic acid (0.084 mL) were added. The mixture was stirred overnight at room temperature. The solvent was evaporated under vacuum, the crude material dissolved in DCM (50 mL) and washed with a saturated NaHCO3 aqueous solution (2×20 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 95\5 to 90\10) to give the title compound (0.872 g, (21S) isomer 95% pure by NMR analysis).
1H-NMR (CDCl3) δ: 7.52 (d, 1H), 7.18 (d, 1H), 7.27 (m, 1H), 7.16 (d, 1H), 7.02 (dd, 1H), 5.58 (dd, 1H), 4.31 (d, 1H), 4.25 (d, 1H), 4.14 (m, 1H), 4.12 (m, 1H), 4.02 (m, 1H), 3.86 (q, 1H), 3.56 (m, 1H), 3.20 (m, 1H), 3.10 (m, 1H), 3.04 (m, 1H), 2.93 (m, 2H), 2.68 (s, —OCH3), 2.60 (m, 1H), 2.50 (m, 1H), 2.32 (m, 1H), 2.30 (s, N(CH3)2), 2.05-1.85 (m, 3H), 1.80 (m, 1H), 1.70 (m, 2H), 1.60 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.34 (s, 3H), 1.32 (d, 3H), 1.25 (d+m, 3H+1H), 1.16 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 111 (0.035 g) in anhydrous acetonitrile (2 mL) was stirred at 50° C. for 6 h. After cooling to room temperature solvent was evaporated, the residue dissolved in anhydrous MeOH (2 mL) and sodium cyanoborohydride (1M in THF, 0.140 mL) and acetic acid (0.010 mL) were added. The mixture was stirred overnight at room temperature. The solvent was evaporated under vacuum, the residue dissolved in DCM (50 mL) and washed with a saturated NaHCO3 aqueous solution (2×20 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 95\5) affording the title compound (0.020 g).
1H-NMR (CDCl3) δ: 7.20-6.80 (m, 3H), 5.66 (dd, 1H), 4.38 (d, 1H), 4.26 (d, 1H), 4.11 (s, 1H), 3.88 (m, 3H), 3.67 (m, 1H), 3.41 (m, 1H), 3.20-2.85 (m, 5H), 2.71 (s, 3H), 2.70 (bs, 6H), 2.59 (m, 1H), 2.41-2.35 (m, 1H+3H), 2.00-1.55 (m, 7H), 1.50 (s, 3H), 1.39 (d, 1H+3H), 1.32-1.27 (m, 3*3H), 1.18 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
A solution of intermediate 112 (0.060 g) in acetonitrile (3 mL) and a 3M HCl aqueous solution (3 mL) was stirred at 80° C. for 3 days. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.050 g) dissolved in anhydrous acetonitrile (2 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 24 h. Acetic acid was added to the mixture to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.070 mL). The reaction mixture was stirred for 16 h at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the product was extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was dissolved in MeOH (1 mL) and stirred overnight. After evaporating the solvent the compound was purified by flash chromatography (eluting with DCM\MeOH from 100\0 to 97\3) to give the title compound (0.004 g).
m\z ([MH]+)=798
A solution of example 7 (0.050 g) and intermediate 113 (0.046 g) in anhydrous acetonitrile (2 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the residue dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.070 mL) and acetic acid (0.009 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give the title compound (0.001 g).
1H-NMR (CDCl3) δ: 8.23 (s, 1H), 5.51 (dd, 1H), 4.38-4.20 (m, 4H), 4.12(s, 1H), 3.86 (m, 1H), 3.55 (m, 1H), 3.18 (dd, 1H), 3.12÷3.00 (m, 2H), 2.98÷2.84 (m, 2H), 2.68 (s, 3H), 2.58 (m, 1H), 2.52 (s, 3H), 2.44 (m, 1H), 2,37 (s, 3H), 2.32 (s, 1H), 2.27 (s, 6H), 2.12 (m, 1H), 2.02 (m, 1H), 1.92 (m, 1H), 1.81 (m, 1H), 1.74÷1.63 (m, 2H), 1.55 (m, 1H), 1.49 (s, 3H), 1.38 (d, 3H), 1.33 (s, 3H), 1.32 (d, 3H), 1.25(m, 1H), 1.25 (d, 3H), 1.15 (d, 3H), 1.09 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 114 (0.041 g) in anhydrous acetonitrile (2 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the residue dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.070 mL) and acetic acid (0.009 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give the title compound (0.009 g).
1H-NMR (CDCl3) δ: 8.79 (d, 1H), 8.07 (d, 1H), 7.97 (d, 1H), 7.79 (d, 1H), 7.65 (t, 1H), 7.52 (t, 1H), 5.74 (dd, 1H), 4.32 (d, 1H), 4.25 (d, 1H), 4.17 (s, 1H), 3.85 (q, 1H), 3.55 (m, 1H), 3.22 (m, 1H), 3.11 (m, 1H), 3.04 (m, 1H), 3.02÷2.78 (m, 4H), 2.69 (s, 3H), 2.99 (m, 1H), 2.52 (m, 1H), 2.34 (s, 1H), 2.32 (s, 6H), 1.94 (m, 1H), 1.89 (m, 2H), 1.88÷1.66 (m, 3H), 1.62÷1.50 (m, 1H), 1.49 (s, 3H), 1.39 (d, 3H), 1.32 (m, 6H), 1.25 (m, 1H), 1.25 (d, 3H), 1.16 (d, 3H), 1.10 (d, 3H), 0.88 (t, 3H).
A solution of example 7 (0.050 g) and intermediate 115 (0.026 g) in anhydrous acetonitrile (1 mL) was stirred at room temperature for 18 h under nitrogen atmosphere. The solvent was evaporated under reduced pressure, the crude dissolved in anhydrous MeOH (2 mL) then sodium cyanoborohydride (1M in THF, 0.037 mL) and acetic acid (0.004 mL) were added. After 18 h the solvent was removed under reduced pressure, the residue dissolved in DCM (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×3 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 94\6) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 8.62 (t, 1H), 8.17 (d, 1H), 8.05 (d, 1H), 7.60 (d, 1H), 4.15 (m, 2H), 4.09 (m, 1H), 4.05 (m, 1H), 3.02 (m, 1H), 3.04-2.92 (m, 2H), 2.33 (m, 1H), 2.05 (m, 1H), 1.98 (m, 1H), 1.12 (d, 3H).
To a solution of example 7 (0.100 g) in anhydrous acetonitrile intermediate 117 (0.066 g) was added portionwise at room temperature under nitrogen atmosphere. After stirring overnight the solvent was removed under reduced pressure, the residue dissolved in MeOH (2.5 mL) and sodium cyanoborohydride (1M in THF, 0.022 mL) and acetic acid (0.013 mL) added. The mixture was stirred overnight. The solvent was evaporated under vacuum, the residue dissolved in EtOAc (5 mL) and washed with a saturated NaHCO3 aqueous solution (2×2 mL) and brine (2 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 97\3) affording the title compound (0.070 g).
m\z ([MH]+)=798
A solution of intermediate 118 (0.065 g) in acetonitrile (2.5 mL) and a 3M HCl aqueous solution (2.5 mL) was stirred at 60° C. for 8 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.040 g) dissolved in anhydrous acetonitrile (1 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The mixture was stirred at room temperature for 24 h. Acetic acid was added to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.060 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material purified by flash chromatography (eluting with DCM\MeOH from 100\0 to 95\5) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 7.66 (d, 1H), 7.25 (d, 1H), 6.62 (m, 1H), 6.51 (d, 1H), 4.40 (m, 1H), 4.30 (m, 1H), 4.15 (m, 1H), 4.00 (s, 3H), 3.03 (m, 1H), 3.05-2.88 (m, 2H), 2.30 (m, 1H), 2.06 (m, 1H), 1.96 (m, 1H), 1.10 (d, 3H).
A solution of intermediate 119 (0.015 g) in acetonitrile (1 mL) and a 3M HCl aqueous solution (1 mL) was stirred at room temperature for 16 h and heated to 50° C. for 8 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.033 g) dissolved in anhydrous acetonitrile (1 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The solution was stirred at room temperature overnight. Acetic acid was added to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.0.25 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The residue was dissolved in MeOH (1 mL) and stirred overnight. After evaporating the solvent the crude material was purified by flash chromatography (eluting with DCM\MeOH from 100\0 to 97\3) and by preparative LC (A\B from 80\20 to 1 0\90 in 20 min) to give the title compound (0.014 g).
1H-NMR (CDCl3) δ: 7.80 (d, 1H), 7.69 (d, 1H), 7.27 (d, 1H), 6.76 (d, 1H), 5.63 (dd, 1H), 4.33 (m, 3H), 4.23 (m, 2H), 3.84 (m, 1H), 3.57 (m, 1H), 3.35 (m, 2H), 3.22 (m, 1H), 3.10-3.00 (m, 2H), 2.64-2.50 (m, 2H), 2.50 (s, 3H), 2.33 (s, 6H), 2.30 (s, 1H), 1.90 (m, 1H), 1.80-1.66 (m, 3H), 1.60-1.40 (m+s, 1H+3H), 1.40-1.20 (m, 4*3H+1H), 1.14 (d, 3H), 1.09 (d, 3H), 0.86 (t, 3H).
A solution of intermediate 120 (0.023 g) in acetonitrile (0.5 mL) and a 3M HCl aqueous solution (0.7 mL) was heated to 70° C. for 24 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.039 g) dissolved in anhydrous acetonitrile (0.5 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 6 h. Acetic acid was added to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.0.58 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the organic solvent the aqueous phase was extracted with EtOAc (3×5 mL). The organic layer was washed with brine (3 mL), dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 97\3 to 95\5) affording the title compound (0.026 g).
1H-NMR (CDCl3) δ: 7.80 (d, 2H), 7.58 (d, 1H), 7.45 (d, 1H), 7.36 (t, 2H), 7.21 (m, 1H), 4.21 (m, 1H), 4.14-4.07 (m, 2H), 3.30-3.20 (m, 2H), 3.07 (m, 1H), 2.29 (m, 1H), 1.110 (d, 3H).
A solution of intermediate 121 (0.088 g) in acetonitrile (1.5 mL) and a 3M HCl aqueous solution (1 mL) was heated to 70° C. for 24 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 7 (0.100 g) dissolved in anhydrous acetonitrile (1 mL) keeping the pH of the solution in the range 6-7 by addition of saturated NaHCO3 aqueous solution. The solution was stirred at room temperature for 6 h. Acetic acid was added to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.150 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the organic solvent the aqueous phase was extracted with EtOAc (3×5 mL). The organic layer was washed with brine (3 mL), dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 97\3 to 95\5). The obtained compound was dissolved in MeOH (5 mL) stirred overnight at room temperature. After evaporating the solvent, the residue was purified by flash chromatography (eluting with DCM\MeOH from 100\0 to 95\5) to give the title compound (0.034 g).
1H-NMR (CDCl3) δ: 7.54 (d, 1H), 7.32 (d, 1H), 7.28 (dd, 1H), 7.15 (dd, 1H), 7.01 (dd, 1H), 4.20 (m, 1H), 4.11-4.07 (m, 2H), 3.30-3.20 (m, 2H), 3.06 (m, 1H), 2.30 (m, 1H), 1.10 (d, 3H).
A solution of example 6 (0.050 g) and quinoline-2-carbaldehyde (0.015 g) in anhydrous toluene (3 mL) was heated to 110° C. for 3 h. The solution was allowed to reach room temperature and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and palladium palladium (10 wt. % on carbon powder, 0.020 g) was added and the mixture stirred under hydrogen atmosphere (1 atm) for 1 h. Filtration through a celite pad eluting with MeOH (10 mL) and purification by flash chromatography (eluting with DCM\MeOH 90\10) gave the title compound (0.009 g).
1H-NMR (CDCl3) δ: 8.74 (s, 1H), 8.30 (d, 1H), 8.21 (d, 1H), 8.13 (td, 1H), 7.83 (t, 1H), 7.57 (t, 1H), 7.30 (t, 1H), 5.16 (bs, 1H), 3.12 (m, 1H), 2.93 (bs, 1H), 1.29 (d, 3H).
A solution of example 6 (0.050 g) and quinoline-3-carbaldehyde (0.016 g) in anhydrous toluene (3 mL) was stirred at 100° C. for 15 h. After evaporation of the solvent under reduced pressure, the residue was dissolved in MeOH (1 mL) and sodium cyanoborohydride (1M in THF, 0.060 mL) and acetic acid (0.004 mL) were added. The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (3×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH 94\6) to give the title compound (0.007 g).
1H-NMR (CDCl3) δ: 8.94 (d, 1H), 8.23 (d, 1H), 8.09 (d, 1H), 7.82 (d, 1H), 7.67 (t, 1H), 7.53 (t, 1H), 4.29 (d, 1H), 4.28 (m, 2H), 4.18 (m, 1H), 3.07 (m, 1H), 2.48 (m, 1H), 1.10 (d, 3H).
To a solution of example 66 (0.010 g) in anhydrous DCM (0.4 mL) DMAP (0.050 g) and acetyl chloride (0.030 mL) were added portionwise over 4 days. Water (2 mL) was added and the mixture was extracted with DCM (3×5 mL). The organic phase was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH from 97\3 to 95\5). The compound was dissolved in MeOH (1 mL) and stirred overnight. Solvent evaporation gave the title compound (0.002 g).
1H-NMR (CDCl3) δ: 8.97(s, 1H), 8.48 (d, 1H), 8.07 (d, 1H), 7.62 (s, 1H), 7.39 (s, 1H), 7.31 (m, 1H), 5.53 (bm, 1H), 4.35 (d, 1H), 4.29 (m, 1H), 4.15 (m, 1H), 4.01 (d, 1H), 3.83 (m, 1H), 3.72 (q, 1H), 3.65 (bs, 1H), 3.57-3.52 (m, 3H), 3.21 (m, 1H), 3.10 (m, 1H), 2.73 (m, 1H), 2.65-2.50 (m, 2H+3H), 2.39-2.22 (m, 2H+6H), 2.00 (s, 3H), 1.99 (m, 1H), 1.70-1.50 (m, 4H), 1.46 (s, 3H), 1.40 (d, 3H), 1.29-1.23 (m, 7H), 1.26 (s, 3H), 1.10 (d, 3H+3H), 0.95 (t, 3H).
To a solution of example 7 (0.070 g) in anhydrous DCM (2 mL) cooled to 0° C. TEA (0.090 mL), DMAP (catalytic amount) and benzyl chloroformate (0.070 g) were added under nitrogen atmosphere. The mixture was stirred at 0° C. for 3 h. After reaching room temperature a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (2×5 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 100\0 to 98\2). The obtained compound was dissolved in MeOH (1 mL) and stirred at room temperature overnight. Solvent evaporation gave the title compound (0.002 g).
1H-NMR (CDCl3) δ: 7.40-7.20 (m, 5H), 5.81 (d, 1H), 5.16 (m, 1H), 5.08 (m, 1H), 4.80 (bm, 1H), 3.07 (m, 1H), 2.44 (m, 1H), 1.18 (d, 3H).
A solution of example 6 (0.050 g) and quinoline-2-carbaldehyde (0.015 g) in anhydrous toluene (3 mL) was heated to 110° C. for 3 h. The solution was allowed to reach room temperature and concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL) and stirred for 16 h. Solvent evaporation gave the title compound (0.050 g).
1H-NMR (CDCl3) δ: 8.99 (d, 1H), 8.47 (d, 1H), 8.09 (d, 1H), 7.58 (s, 1H), 7.41 (s, 1H), 7.30 (m, 1H), 6.73 (d, 1H), 4.88 (dd, 1H), 4.67 (s, 1H), 3.08 (m, 1H), 2.41 (d, 1H), 1.16 (d, 3H).
To a solution of (quinoxalin-2-ylsulfanyl)-acetic acid (0.056 g) in anhydrous DMF (4 mL) under a nitrogen atmosphere HATU (0.097 g) and DIPEA (0.053 mL) were added. The reaction mixture was stirred at room temperature for 20 min then example 11 (0.160 g) was added. The reaction mixture was stirred at room temperature for 6 h then it was diluted with DCM (10 mL) and washed with a 5% NaHCO3 aqueous solution (10 mL). The aqueous phase was extracted with DCM (3×10 mL), the combined organic layers washed with a 5% NaHCO3 aqueous solution (10 mL), dried over Na2SO4 and evaporated under reduced pressure. The residue was dissolved in MeOH (5 mL) and stirred at room temperature overnight. After evaporating the solvent the crude material was purified by flash chromatography (DCM\MeOH 95\5) to give the title compound (0.126 g).
m\z ([MH]+)=847
1H-NMR (CDCl3) δ: 8.63 (s, 1H), 8.20 (d, 1H), 7.99 (m, 2H), 7.72 (t, 1H), 7.65 (t, 1H), 5.47 (dd, 1H), 4.36 (d, 1H), 3.98 (s, 1H), 3.92 (d, 1H), 3.51 (m, 1H), 3.16 (m, 1H), 3.02 (m, 1H), 2.88 (m, 1H), 2.64 (s, 3H), 2.58 (m, 1H), 2.36 (m, 1H), 2.34 (s, 6H2), 2.10 (m, 1H), 2.03 (m, 1H), 1.97 (m, 1H), 1.73 (m, 4H), 1.60-1.40 (m, 8H), 1.23 (m, 4H), 1.15-1.01 (m, 9H), 0.90 (t, 3H).
To a solution of 4-(2-hydroxy-4,5-dimethoxy-phenyl)-4-oxo-butyric acid (0.031 g) in anhydrous DMF (2 mL) HATU (0.047 g) and DIPEA (0.025 mL) were added under nitrogen atmosphere. After stirring for 30 min example 10 (0.070 g) was added and the mixture stirred overnight. A 5% NaHCO3 aqueous solution (3 mL) was added and the mixture extracted with DCM (2×3 mL). The organic phase was dried over Na2SO4 and evaporated under reduced pressure. The residue was dissolved in MeOH (3 mL) and stirred overnight. After solvent evaporation the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 85\15\0.2) to give the title compound (0.027 g).
1H-NMR (CDCl3) δ: 12.46 (s, 1H), 7.15 (s, 1H), 6.72 (d, 1H), 6.44 (s, 1H), 4.44 (m, 1H), 3.91 (s, 3H), 3.34 (s, 3H), 3.34 (m, 2H), 3.04 (m, 1H), 2.70-2.52 (m, 2H), 2.34 (m, 1H), 1.80 (d, 3H), 1.17 (d, 3H).
To a solution of 4-(3,4-dimethoxy-phenyl)-4-oxo-butyric acid (0.029 g) in anhydrous DMF (2 mL) HATU (0.047 g) and DIPEA (0.025 mL) were added under nitrogen atmosphere. After stirring for 30 min example 10 (0.070 g) was added and the mixture stirred overnight. A 5% NaHCO3 aqueous solution (3 mL) was added and the mixture extracted with DCM (2×3 mL). The organic phase was dried over Na2SO4 and evaporated under reduced pressure. The residue was dissolved in MeOH (3 mL) and stirred overnight. After solvent evaporation the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 85\15\0.2) to give the title compound (0.024 g).
1H-NMR (CDCl3) δ: 7.64 (d, 1H), 7.53 (m, 1H), 6.89 (d, 1H), 6.77 (d, 1H), 4.40 (t, 1H), 3.96 (s, 3H), 3.94 (s, 3H), 3.35 (m, 2H), 3.04 (m, 1H), 2.70-2.55 (m, 2H), 2.34 (m, 1H), 1.81 (d, 3H), 1.17 (d, 3H).
To a solution of 4-(4-methoxy-3-nitro-phenyl)-4-oxo-butyric acid (0.031 g) in anhydrous DMF (2 mL) HATU (0.047 g) and DIPEA (0.025 mL) were added under nitrogen atmosphere. After stirring for 30 min example 10 (0.070 g) was added and the mixture stirred overnight. A 5% NaHCO3 aqueous solution (3 mL) was added and the mixture extracted with DCM (2×3 mL). The organic phase was dried over Na2SO4 and evaporated under reduced pressure. The residue was dissolved in MeOH (3 mL) and stirred overnight. After solvent evaporation the crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 85\15\0.2) to give the title compound (0.030 g).
1H-NMR (CDCl3) δ: 8.49 (d, 1H), 8.20 (dd, 1H), 7.15 (d, 1H), 6.66 (d, 1H), 4.40 (t, 1H), 4.04 (s, 3H), 3.31 (m, 2H), 2.99 (m, 1H), 2.73 (m, 1H), 2.61 (m, 1H), 2.18 (m, 1H), 1.18 (d, 3H).
A solution of intermediate 51 (0.070 g) in acetonitrile (2.5 mL) and a 3M HCl aqueous solution (2.5 mL) was heated to 80° C. for 24 h. The reaction mixture was allowed to reach room temperature, then it was added dropwise to a solution of example 10 (0.070 g) dissolved in anhydrous acetonitrile (2 mL) keeping the pH of the solution in the range 6-7 by addition of a saturated NaHCO3 aqueous solution. The mixture was stirred at room temperature overnight. Acetic acid was added to reach pH 5-6 followed by sodium cyanoborohydride (1M in THF, 0.150 mL). The reaction mixture was stirred overnight at room temperature. After evaporating the solvent a saturated NaHCO3 aqueous solution (3 mL) was added and the mixture was extracted with DCM (2×5 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by flash chromatography (eluting with DCM\MeOH\NH4OH from 100\0\0 to 80\20\0.2) to give the title compound (0.006 g).
1H-NMR (CDCl3) δ: 9.05 (d, 1H), 8.44 (d, 1H), 8.13 (m, 1H), 7.65 (d, 1H), 7.57 (d, 1H), 7.27 (m, 2H), 4.20-4.10 (m, 2H), 4.07 (bs, 1H), 3.40-3.25 (m, 2H), 2.87 (m, 1H), 2.32 (m, 1H), 1.81 (d, 3H), 1.08 (d, 3H).
A solution of example 10 (0.070 g) and intermediate 52 (0.035 g) in anhydrous acetonitrile (3.5 mL) was stirred at room temperature for 6 h. After evaporating the solvent the residue was dissolved in anhydrous MeOH (3 mL) and sodium cyanoborohydride (1M in THF, 0.051 mL) and acetic acid (0.008 mL) were added. The mixture was stirred overnight. The solvent was evaporated under vacuum, the residue dissolved in DCM (10 mL) and washed with a saturated NaHCO3 aqueous solution (2×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH\NH4OH from 100\0\0 to 80\20\0.2) to give the title compound (0.012 g).
m\z ([MH]+)=830.
1H-NMR (CDCl3) δ: 8.99 (d, 1H), 8.46 (dd, 1H), 8.11 (dd, 1H), 7.63 (s, 1H), 7.40 (s, 1H), 7.30 (m, 1H), 5.45 (dd, 1H), 4.42 (d, 1H), 4.18 (m, 1H), 4.08 (m, 1H), 4.04 (m, 1H), 3.55 (m, 1H), 3.18 (dd, 1H), 3.11 (m, 1H), 3.00 (m, 1H), 2.92 (m, 1H), 2.95 (s, 3H), 2.95 (m, 1H), 2.58 (m, 1H), 2.52 (m, 1H), 2.30 (s, 6H), 1.95 (m, 2H), 1.83 (d, 3H), 1.57 (d, 3H), 1.7 (m, 1H), 1.68 (m, 1H), 1.60 (m, 1H), 1.33 (s, 3H), 1.26 (s, 3H), 1.25 (m, 1H), ), 1.25 (d, 3H), 1.16 (d, 3H), 1.09 (d, 3H), 0.92 (t, 3H).
To a solution of example 13 (0.039 g) in anhydrous DCM (2 mL) cooled to −10° C. a solution of 6-methyl-2-nitroisocyanate (0.010 g) in anhydrous DCM (2 mL) was added and the mixture was stirred at −10° C. for 2 h. The reaction mixture was quenched with a saturated NaHCO3 aqueous solution (2 mL), the aqueous phase was extracted with DCM (2 mL). The organic phase was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH from 97\3 to 95\5) to give the title compound (0.010 g).
1H-NMR (CDCl3) δ: 7.97 (bs, 1H), 7.78 (d, 1H), 7.44 (d, 1H), 7.15 (t, 1H), 6.23 (bs, 1H), 4.22 (s, 1H), 3.45 (m, 1H), 3.26-3.19 (m, 2H), 3.08 (m, 1H), 2.91 (m, 1H), 2.32 (s, 1H), 1.12 (d, 3H).
To a solution of intermediate 38 (0.33 g) in DCM (30 mL) Dess-Martin periodinane (0.300 g) was added portionwise within 3 h. A Na2S2O3 solution (5% in a saturated NaHCO3 aqueous solution, 20 mL) was added and the mixture was stirred for 1 h. The aqueous phase was extracted with DCM (2×50 mL), the organic phase was washed with water (2×30 mL), dried over Na2SO4 and evaporated under reduced pressure. Purification of the crude material by flash chromatography (eluting with: DCM\MeOH\NH3 9.6\0.3\0.09) gave the title compound (0.13 g).
1H-NMR (CDCl3) δ: 5.69 (m, 1H), 5.43 (dd, 1H), 5.09 (m, 2H), 4.75 (m, 1H), 4.73 (s, 1H), 4.47 (d, 1H), 4.39 (d, 1H), 3.91 (q, 1H), 3.70 (m, 2H), 3.63 (m, 1H), 3.21 (m, 1H), 3.20 (s, 1H), 3.12 (m, 1H), 2.70 (m, 1H), 2.65 (m, 1H), 2.26 (s, 6H), 2.04 (s, 3H), 1.94 (m, 1H), 1.70 (m, 1H), 1.65 (m, 1H), 1.60 (s, 3H), 1.55 (m, 1H), 1.36 (d, 3H), 1.33 (s, 3H), 1.26 (d, 6H), 1.13 (d, 3H), 1.06 (d, 3H), 0.93 (t, 3H).
A solution of example 140 (0.005 g) in MeOH (0.5 mL) was stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.003 g).
1H-NMR (CDCl3) δ: 5.71 (m, 1H), 5.44 (dd, 1H), 5.08 (m, 2H), 4.72 (s, 1H), 4.43 (d, 1H), 4.40 (d, 1H), 3.94 (q, 1H), 3.72 (m, 2H), 3.62 (m, 1H), 3.20 (m, 4H), 2.68 (m, 1H), 2.52 (m, 1H), 2.29 (s, 6H), 1.94 (m, 1H), 1.81 (m, 1H), 1.65 (m, 1H), 1.60 (m, 1H), 1.61 (s, 3H), 1.42 (d, 3H), 1.35 (s, 3H), 1.34 (d, 3H), 1.26 (d, 3H), 1.12 (d, 3H), 1.08 (d, 3H), 0.93 (t, 3H).
To a solution of intermediate 42 (0.528 g) and EDC (0.35 g) in DCM anhydrous (40 mL) cooled to 0° C., DMSO (0.4 mL) was added. After 10 min at 0° C., a solution of pyridinium trifluoroacetate (0.36 g) in DCM (2 mL) was slowly added. After 10 min the ice bath was removed. Two further additions of EDC (0.35 g each time), DMSO (0.4 mL each time) and pyridinium trifluoroacetate (0.36 g each time) were performed. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3×50 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum to give the title compound (0.520 g).
TLC: DCM\MeOH\NH3 20\2\0.2 (Rf=0.39).
A solution of example 142 (0.52 g) in acetonitrile (66 mL) and 1.2N HCl aqueous solution (154 mL) was stirred at room temperature for 1 h. After neutralising the mixture with solid Na2CO3 and evaporating the solvent under vacuum, the mixture was extracted with DCM (3×50 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give the title compound (0.47 g).
1H-NMR (CDCl3) δ: 5.88 (dd, 1H), 5.70 (m, 1H), 5.13 (d, 1H), 4.75 (m, 1H), 4.57 (s, 1H), 4.44 (d, 1H), 4.38 (d, 1H), 3.91 (q, 1H), 3.75 (q, dd), 3.61 (m, 1H), 3.50 (m, 1H), 3.24 (m, 1H), 3.08 (m, 1H), 2.70 (m, 1H), 2.64 (m, 1H), 2.46 (bs, 1H), 2.26 (s, 6H), 2.18 (m, 6H), 1.60-1.40 (m, 5H), 1.40-1.20 (m, 13H), 1.15 (d, 3H), 1.08 (d, 3H), 0.88 (t, 3H).
A solution of example 143 (0.002 g) in MeOH (0.3 mL) was stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.002 g).
1H-NMR (CDCl3) δ: 5.82 (dd, 1H), 5.73 (m, 1H), 5.14 (d, 1H), 5.02 (d, 1H), 4.54 (s, 1H), 4.40 (d, 1H), 4.38 (d, 1H), 3.93 (q, 1H), 3.76 (m, 1H), 3.62 (m, 1H), 3.52 (m, 1H), 3.24 (m, 1H), 3.21 (m, 1H), 3.09 (m, 1H), 2.66 (m, 1H), 2.50 (m, 1H), 2.47 (m, 1H), 2.28 (s, 6H), 1.95 (m, 1H), 1.85 (m, 2H), 1.80-0.80 (several m, 27H).
m\z ([MH]+)=653.
To a solution of quinolin-2-thiol (0.008 g) in anhydrous DMF (0.500 mL) sodium hydride (1.2 mg) was added and the mixture stirred at room temperature for 15 min then intermediate 24 (0.025 g) was added and the reaction mixture stirred at 60° C. for 5 h. The reaction mixture was diluted with DCM (3 mL) and washed with a saturated NaHCO3 aqueous solution (1 mL). The organic phase was dried over Na2SO4 and evaporated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 98\2) to give a compound that was dissolved in MeOH (1 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.010 g).
LC\MS analysis (mobile phase: A\B from 90\10 to 10\90 in 10 min, 10\90 for 2 min, mass range 150-1300 amu): retention time: 8.7 min, m\z ([MH]+)=815.
To a solution of benzothiazole-2-thiol (0.009 g) in anhydrous DMF (0.500 mL) sodium hydride (1.2 mg) was added and the mixture stirred at room temperature for 15 min then intermediate 24 (0.025 g) was added and the reaction mixture stirred at 60° C. for 5 h. The reaction mixture was diluted with DCM (3 mL), washed with a saturated NaHCO3 aqueous solution (1 mL). The organic phase was dried over Na2SO4 and evaporated under reduced pressure. The crude material was purified by flash chromatography (eluting with: DCM\MeOH 98\2) to give a compound that was dissolved in MeOH (1 mL) and stirred at room temperature overnight. Solvent evaporation under reduced pressure gave the title compound (0.012 g).
LC\MS analysis (mobile phase: A\B from 90\10 to 10\90 in 10 min, 10\90 for 2 min, mass range 150-1300 amu): retention time: 8.5 min, m\z ([MH]+)=821.
To 3-pyridin-3-yl-acrylic acid (1.3 mg) a solution of HATU (0.003 g) in anhydrous DMF (0.050 mL) and DIPEA (0.002 mL) in anhydrous DMF (0.050 mL) were added, followed by a solution of example 6 (0.005 g) in anhydrous DMF (0.050 mL). The reaction mixture was stirred at room temperature for 18 h, then it was diluted with DCM (0.350 mL), washed with a 5% NaHCO3 aqueous solution (0.300 mL), then passed through a phase-separation syringe. The aqueous phase was extracted with DCM (0.250 mL) and the collected organic extracts evaporated under vacuum. The crude material was dissolved in DCM (0.700 mL), loaded on SCX-cartridge (250 mg, loading 0.28 mmol/g, previously washed with 4 mL of MeOH), washed with MeOH (3.5 mL), then the product eluted with NH3 (0.25M solution in MeOH, 1 mL), followed by MeOH (0.7 mL). The collected fractions were left in the NH3/MeOH solution overnight. After evaporating the solvent the title compound (0.002 g) was obtained.
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 10 min, 10/90 for 5 min; mass range 150-1200 amu): retention time: 5 min, m\z ([MH]+)=758.
Examples 148-312 were obtained starting from example 6 (5 mg) by following the same procedure as reported for Example 147.
The name and amount of starting material (i.e carboxylic acid) and LC/MS analysis (retention time and m/z) of examples 148-312 are reported in the Table 1.
To (2R)-tert-butoxycarbonylamino-3-(1H-indol-3-yl)-propionic acid (0.0025 g) a solution of HATU (0.003 g) in anhydrous DMF (0.050 mL) and DIPEA (0.002 mL) in anhydrous DMF (0.050 mL) were added, followed by a solution of example 6 (0.005 g) in anhydrous DMF (0.050 mL). The reaction mixture was stirred at room temperature for 18 h, then it was diluted with DCM (0.350 mL), washed with a 5% NaHCO3 aqueous solution (0.300 mL), then passed through a phase-separation syringe. The aqueous phase was extracted with DCM (0.250 mL) and the collected organic extracts evaporated under vacuum. The residue was dissolved in a 10% TFA solution in anhydrous DCM (0.300 mL) and the mixture stirred for 1.5 h. The solution was diluted with EtOAc (0.400 mL) then solvents evaporated under reduced pressure.
The crude material was dissolved in DCM (0.700 mL), loaded on SCX-cartridge (100 mg, loading 0.75 mmol/g, previously washed with 4 mL of MeOH), washed with MeOH (4 mL), then the product eluted with NH3 (0.25M solution in MeOH, 1.5 mL), followed by MeOH (2 mL) and solvents evaporating under vacuum. The residue was dissolved in MeOH (1.7 mL) and stirred overnight at room temperature. After evaporating the solvent the title compound (0.001 g) was obtained.
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 10 min, 10/90 for 3 min; mass range 150-1000 amu): retention time: 4.95 min, m\z ([MH]+)=813.
Examples 314-320 were obtained starting from example 6 (5 mg) by following the same procedure as reported for Example 313.
The name and amount of starting material (i.e carboxylic acid) and LC/MS analysis (retention time and m/z) of examples 314-320 are reported in the table 2.
To (2S)-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}-3-pyridin-4-yl-propionic acid (0.0032 g) a solution of HATU (0.003 g) in anhydrous DMF (0.050 mL) and DIPEA (0.002 mL) in anhydrous DMF (0.050 mL) were added, followed by a solution of example 6 (0.005 g) in anhydrous DMF (0.050 mL). The reaction mixture was stirred at room temperature for 18 h, then it was diluted with DCM (0.350 mL), washed with a 5% NaHCO3 aqueous solution (0.300 mL), then passed through a phase-separation syringe. The aqueous phase was extracted with DCM (0.250 mL) and the collected organic extracts evaporated under vacuum. The residue was dissolved in anhydrous DMF (0.350 mL) then piperazinomethyl polystyrene resin (0.030 g, loading 1.39 mmol/g) was added and the mixture stirred for 2.5 days. The mixture was filtered and the resin rinsed with DCM (0.400 mL), DMF (0.400 mL) and DCM (0.200 mL) and the filtrates evaporated under reduced pressure.
The crude material was dissolved in DCM (0.700 mL), loaded on SCX-cartridge (100 mg, loading 0.75 mmol/g, previously washed with 4 mL of MeOH), washed with MeOH (4 mL), then the product eluted with NH3 (0.25M solution in MeOH, 1.5 mL), followed by MeOH (2 mL) and solvents evaporating under vacuum. The residue was dissolved in MeOH (1.7 mL) and stirred overnight at room temperature. After evaporating the solvent the title compound (0.001 g) was obtained.
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 10 min, 10/90 for 3 min; mass range 150-1000 amu): retention time: 3.98/4.70 min, m\z ([MH]+)=775.
Examples 322-327 were obtained starting from example 6 (5 mg) by following the same procedure as reported for Example 321.
The name and amount of starting material (i.e carboxylic acid) and LC/MS analysis (retention time and m/z) of examples 322-327 are reported in the table 3.
To a solution of example 6 (0.005 g) in anhydrous THF (0.100 mL) a solution of benzoyl isocyanate (2.2 mg) in anhydrous THF (0.300 mL) was added. The reaction mixture was heated at 60° C. for 24 h. After cooling to room temperature PS-Trisamine resin (loading 3.62 mmol/g, 0.030 g) was added and reacted at 60° C. for 24 h. After cooling to room temperature the mixture was filtered and the resin rinsed with THF (2×0.240 mL), DCM (2×0.240 mL), THF (4×0.170 mL) and the filtrate evaporated. The residue was dissolved in MeOH (1 mL) and reacted at room temperature overnight. After evaporating the solvent the title compound (0.003 g) was obtained.
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 10 min, 10/90 for 2 min; retention time: 6.08/6.51 min, m\z ([MH]+)=774.
Examples 329-398 were obtained starting from example 6 (5 mg) by following the same procedure as reported for Example 328.
The name and amount of starting material (i.e isocyanate) and LC/MS analysis (retention time and m/z) of examples 329-398 are reported in the table 4.
A solution of example 6 (0.005g) in MeOH (0.350 mL) was reacted overnight at room temperature. After evaporating the solvent a solution of benzoyl isothiocyanate (2.4 mg) in DCE (0.400 mL) was added and the reaction mixture was heated at 60° C. for 26 h. After cooling to room temperature PS-Trisamine resin (loading 3.62 mmol/g, 0.030 g) was added and reacted at room temperature for 15 h. The mixture was filtered and the resin rinsed with DCE (2×0.230mL), DCM (3×0.160mL, 2×0.120mL). The filtrate was evaporated to give the title compound (0.003 g).
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 16 min, 10/90 for 4 min; retention time: 9.1/10.0 min, m\z ([MH]1)=790.
Examples 400-425 were obtained starting from example 6 (5 mg) by following the same procedure as reported for Example 399.
The name and amount of starting material (i.e isocyanate) and LC/MS analysis (retention time) of examples 400-425 are reported in the table 5.
To 3-amino-isonicotinic acid (0.001 g) a solution of benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (0.004 g) in anhydrous DMF (0.150 mL) and a solution of DIPEA (0.003 mL) in anhydrous DMF (0.150 mL) were added followed by the addition of a solution of example 13 (0.005 g) in anhydrous DMF (0.100 mL). The reaction mixture was stirred at room temperature for 48 h, then it was diluted with DCM (0.600 mL), washed with a 5% NaHCO3 aqueous solution (0.500 mL), then passed through a phase-separation syringe. The aqueous phase was extracted with DCM (0.400 mL) and the collected organic extracts evaporated under vacuum. The crude material was dissolved in DCM (0.500 mL), loaded on a SCX-cartridge (250 mg, loading 0.75 mmol/g), washed with MeOH (4 mL), then the product eluted with NH3 (0.25M solution in MeOH, 1.5 mL). After evaporating the solvent the title compound (0.003 g) was obtained.
LC/MS analysis (mobile phase: A/B from 90/10 to 10/90 in 10 min, 10/90 for 2 min, mass range 150-1300 amu): retention time: 4.99 min, m/z ([MH]+)=790.
Example 427-601 were obtained starting from example 13 (5 mg) by following the same procedure as reported for Example 426.
The name and amount of starting material (i.e carboxylic acid) and LC/MS analysis (retention time and m/z) of examples 427-601 are reported in table 6.
Tablets
The active ingredient and the lactose are blended together and then granulated using water as granulating fluid. The dried granules are blended with ethyl cellulose, sodium lauryl sulphate and magnesium stearate and the tablet core formed using an appropriate punch. The tablet may be coated using conventional technique and coatings.
Injection
The sterile vials were filled with the sterile active ingredient (500 mg). Purge the vial head space with sterile nitrogen; close the vials using rubber and metal overseals. The product may be constituted by dissolving in water for injection(10 ml) or other suitable sterile vehicle for injection shortly before administration.
Activity Data
The value of MIC (microbial inhibition concentration), obtained according to NCCLS (National Committee for Clinical Laboratory Standards), of the preferred compounds of the invention against erythromycin susceptible Streptococcus pneumoniae and Streptococcus pyogenes are less then or equal to 1 ug/ml.
In particular Examples 70, 56, 54, 49, 51, 58, 75, 66, 47, 52, 117 showed MIC<=0.1 ug/ml against erythromycin susceptible Streptococcus pneumoniae strains. Furthermore Examples 70, 56, 54, 49, 51, 58, 75, 66, 47, 52, 117 showed MIC in the range <=8-0.06 ug/ml against erythromycin resistant Streptococcus pneumoniae strains.
| Number | Date | Country | Kind |
|---|---|---|---|
| 0031309.8 | Dec 2000 | GB | national |
| 0126277.3 | Nov 2001 | GB | national |
| 0126276.5 | Nov 2001 | GB | national |
This Application is a continuation of application Ser. No. 11/127,701, filed May 12, 2005, which is a continuation of application Ser. No. 10/450,893, filed Nov. 19, 2003 (now abandoned), which is a 371 of International Application No. PCT/GB01/05665, filed Dec. 20, 2001.
| Number | Date | Country | |
|---|---|---|---|
| Parent | 11127701 | May 2005 | US |
| Child | 11422122 | Jun 2006 | US |
| Parent | 10450893 | Nov 2003 | US |
| Child | 11127701 | May 2005 | US |
