Patent Application
20220073475
The present invention relates to novel di-substituted 5-membered heteroaryl compounds containing a hydroxamate moiety including their pharmaceutically acceptable salts and solvates which are useful as sensitizers for chemotherapy of malignant tumors, particularly in multiple myeloma, cervical cancer and breast cancer, and are useful as therapeutic compounds, particularly in the treatment and/or prevention of cancers that may be treated by cytotoxic agents, such as proteasome inhibitors, histone deacetylase (HDAC) inhibitors and/or protein glycosylation inhibitors.
Chemotherapy is a type of treatment that includes a medication to treat cancer. The goal of chemotherapy is to stop or slow the growth of cancer cells. Chemotherapy medications attack rapidly growing cancer cells, but they can also affect healthy cells that grow rapidly. The effect of these medications on normal cells often causes side effects.
Histone deacetylase (HDAC) is a kind of metalloprotease, which plays a key role in chromosome structure modification and gene expression modulation. In recent years, it has been found that inhibiting HDACs activity can effectively inhibit cancer cell proliferation, induce cell cycle arrest and promote cell apoptosis, and therefore, HDACs have become a new target of anticancer drug design, and using HDAC inhibitor is considered as an effective strategy for cancer therapy. Particularly, hydroxamic acid compounds have been paid much attention and demonstrated a good anti-tumor activity, both in vitro and in vivo. In 2006, vorinostat was the first hydroxamic acid compound approved by the United States FDA and is used for treating skin T cell lymphoma. Other examples of hydroxamic acid compounds having an activity as HDAC inhibitors and their use for the treatment of cancer are namely disclosed in WO 2010/151318 A1, WO 2012/098132 A1, CN 107445896 A and US 2016/176879 A1.
Proteasomes are protein complexes which degrade unneeded or damaged proteins by proteolysis, a chemical reaction that breaks peptide bonds. Proteasome inhibitors are drugs that block the action of proteasomes, and are being studied in the treatment of cancer; and three are approved for use in treating multiple myeloma. Multiple mechanisms are likely to be involved, but proteasome inhibition may prevent degradation of pro-apoptotic factors such as the p53 protein, permitting activation of programmed cell death in neoplastic cells dependent upon suppression of pro-apoptotic pathways. For example, bortezomib causes a rapid and dramatic change in the levels of intracellular peptides.
Most of anticancer agents are used in combination. Synergistic effect is searched for a better efficacy and lowering of resistance. Also, sensitizers are used to effectively increase the sensitivity of tumor cells to treatments under a lower dose.
U.S. Pat. No. 9,126,952 B2 discloses morpholinylquinazoline compounds and their use for the sensitization of cancer cells to anticancer agents.
However, there is still a need for new compounds having the ability to sensitize cancer cells to various anticancer drugs, and that may be of therapeutic value in the treatment of cancers that may be treated by cytotoxic agents, such as proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors.
The inventors have now succeeded in developing novel compounds based on a 5-membered heteroaryl scaffold bearing two side groups, one of which carries a hydroxamic acid moiety. These compounds have the advantage of increasing the cytotoxicity of various anticancer drugs.
The invention therefore relates to compounds of general Formula I, their pharmaceutically acceptable salts and solvates as well as methods of use of such compounds or compositions comprising such compounds as sensitizers for chemotherapy of malignant tumors.
In a general aspect, the invention provides compounds of general Formula I:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1 is inexistent or is selected from —CH2— and —CH(OH)—;
R1 is selected from aryl, heteroaryl, arylalkyl and arylalkenyl wherein said aryl, heteroaryl, arylalkyl or arylalkenyl is optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen, C1-C4-haloalkyl, hydroxy-C1-C4-alkyl, acetylamino, acetylamino-C1-C4-alkyl and C1-C4-alkylaminocarboxyl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl, C1-C6-alkyl, C1-C4-alkyloxycarbonyl, C1-C4-alkoxy, arylamino, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl, C1-C4-alkyl, C1-C2-alkoxy-C1-C2-alkoxy, C1-C4-alkyloxycarbonyl, acetylamino, di(C1-C4-alkyl)amino-C1-C4-alkyl and hydroxycarbamoyl;
X is selected from
R3 is selected from H and C(O)OR4;
R4 is C1-C4-alkyl;
L2 is linear or branched C1-C6-alkyl, optionally substituted by a group R5;
R5 is selected from linear or branched C1-C4-alkyl, CH2—OH, CHOH—CH3, CH2—C(O)NH2, CH2—CH2—C(O)NH2, CH2—COOH, CH2—CH2—COOH, (CH2)4—NH2, (CH2)4—NH—C(NH2+)—NH2, CH2—Imidazolyl, CH2—Indolyl, CH2—SH, CH2—CH2—S—CH3, CH2-Ph, CH2-Ph-OH, CH2—OR6, CH2—COOR6, CH2—CH2—COOR6, CH2—SR6 and CH2-Ph-OR6;
R6 is selected from Me, Bn, Ph and Ac;
Z is selected from —NR7(CO)—, —NHSO2—, —CH2—CH2—, —NH—, —NH—CH(C1-C6-alkyl)—, —O—CH2—, —C(O)NH— and —O—; and
R7 is selected from H, linear or branched C1-C6-alkyl and (1-methylimidazol-2-yl)-C1-C2-alkyl, or
R7 and L2 form together with the nitrogen atom they are attached to a 5- or 6-membered heterocyclyl group;
with the proviso that the compound of Formula I is none of the following:
In another aspect, the present invention provides a pharmaceutical composition comprising at least one compound according to the invention or a pharmaceutically acceptable salt or solvate thereof and at least one pharmaceutically acceptable carrier, diluent, excipient and/or adjuvant.
The invention also relates to the use of compounds of Formula I or their pharmaceutically acceptable salts and solvates for the treatment and/or prevention of cancer, particularly as sensitizers for chemotherapy of malignant tumors. Thus, the compound of the invention is useful for the treatment and/or prevention of cancers that may be treated by cytotoxic agents, such as proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors.
As detailed above, the invention relates to compounds of Formula I, as well as their pharmaceutically acceptable salts and solvates.
Preferred compounds of Formula I and pharmaceutically acceptable salts and solvates thereof are those wherein one or more of L1, L2, X, Y, Z, R1, R2, R3, R4, R5, R6 and R7 are defined as follows:
L1 is —CH2— or —CH(OH)—; more preferably L1 is —CH2—;
A is CH or N; preferably A is CH;
Y is —CH2— or —S—; preferably Y is —CH2—;
R1 is selected from naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 5-indolyl, 6-indolyl, 3-quinolinyl, 4-quinolinyl, 7-quinolinyl, dihydroquinolin-2-on-6-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl, styryl, 5-benzothiophenyl, 6-benzothiophenyl, 3-benzothiophenyl, 2-benzothiophenyl, 6-(1,3-benzothiazolyl and 6-benzodioxinyl, wherein said naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 5-indolyl, 6-indolyl, 3-quinolinyl, 4-quinolinyl, 7-quinolinyl, dihydroquinolin-2-on-6-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl, styryl, 5-benzothiophenyl, 6-benzothiophenyl, 3-benzothiophenyl, 2-benzothiophenyl, 6-(1,3-benzothiazolyl or 6-benzodioxinyl is optionally substituted by one or more substituents selected from C1-C4-alkyl, halo, preferably bromo or chloro, more preferably bromo, C1-C4-haloalkyl, preferably C1-C4-fluroalkyl, hydroxy-C1-C4-alkyl, preferably hydroxymethyl, acetylamino, acetylamino-C1-C4-alkyl, preferably acetylaminomethyl, and C1-C4-alkylaminocarboxyl, preferably methylaminocarboxyl; preferably R1 is selected from naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 3-quinolinyl, 4-quinolinyl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl and styryl, wherein said naphthalen-2-yl, phenyl, biphenyl, tetraisoquinolinyl, 3-indolyl, 3-quinolinyl, 4-quinolinyl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl or styryl is optionally substituted by one substituent selected from C1-C4-alkyl, halo, preferably bromo or chloro, more preferably bromo, C1-C4-haloalkyl, preferably C1-C4-fluroalkyl, hydroxy-C1-C4-alkyl, preferably hydroxymethyl, acetylamino, acetylamino-C1-C4-alkyl, preferably acetylaminomethyl, and C1-C4-alkylaminocarboxyl, preferably methylaminocarboxyl; more preferably R1 is selected from naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 3-quinolinyl, 4-quinolinyl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl and styryl wherein said naphthalen-2-yl, phenyl, biphenyl or (2H-1,2,3,4-tetrazol-5-yl)phenyl is optionally substituted by one substituent selected from halo, preferably bromo or chloro, more preferably bromo, C1-C4-haloalkyl, preferably C1-C4-fluroalkyl, more preferably trifluoromethyl, hydroxy-C1-C4-alkyl, preferably hydroxymethyl, acetylamino, acetylamino-C1-C4-alkyl, preferably acetylaminomethyl, and C1-C4-alkylaminocarboxyl, preferably methylaminocarboxyl;
R2 is selected from phenyl, 1,4-imidazolyl, pyrimidinyl, pyridinyl, phenyl-C1-C2-alkyl, cyclohexyl, piperidinyl, morpholinyl, 1,3-benzodioxolyl, C1-05-alkyl, t-butyloxycarbonyl, t-butyloxy, phenylamino, wherein said phenyl, 1,4-imidazolyl, phenyl-C1-C2-alkyl, piperidinyl, is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, preferably methoxy, halogen, preferably fluoro and chloro, C1-C4-haloalkyl, preferably trifluoromethyl, C1-C4-alkyl, preferably methyl, C1-C2-alkoxy-C1-C2-alkoxy, preferably methoxyethoxy, C1-C4-alkyloxycarbonyl, preferably t-butyloxycarbonyl, acetylamino, di(C1-C4-alkyl)amino-C1-C4-alkyl, preferably dimethylaminomethyl, and hydroxycarbamoyl.
X is selected from
more preferably X is
even more preferably X is
R3 is H or C(O)OR4; preferably R3 is H;
R4 is C1-C2-alkyl; more preferably R4 is ethyl;
L2 is selected from —CH2—, —CH2—CH2—, —C(CH3)2— and —CH(R5)—; more preferably L2 is selected from —CH2—, —CH2—CH2— and —CH(R5)—; even more preferably L2 is —CH2—;
R5 is selected from CH3, CH(CH3)—CH3, CH2—CH(CH3)—CH3, CH(CH3)—CH2—CH3, CH2—OH, CHOH—CH3, CH2—C(O)NH2, CH2—CH2—C(O)NH2, CH2—COOH, CH2—CH2—COOH, (CH2)4—NH2, (CH2)4—NH—C(NH2+)—NH2, CH2—Imidazolyl, CH2—Indolyl, CH2—SH, CH2—CH2—S—CH3, CH2-Ph, CH2-Ph-OH, CH2—OR6, CH2—COOR6, CH2—CH2—COOR6, CH2—SR6 and CH2-Ph-OR6; more preferably R5 is selected from CH3, CH2—OH, CH2-Ph-OH and CH2-Ph-OR6; even more preferably R5 is CH3;
R6 is selected from Me, Bn, Ph and Ac, more preferably R6 is Bn;
Z is selected from —NR7(CO)—, —NHSO2— and —C(O)NH—; more preferably Z is —NR7(CO)—; and
R7 is selected from H, methyl, ethyl, isobutyl, isopentyl and (1-methylimidazol-2-yl)methyl; more preferably R7 is H; or
R7 and L2 form together with the nitrogen atom they are attached to a 5- or 6-membered heterocyclyl group selected from pyrrolidinyl, morpholinyl, thiazolidinyl and piperidinyl; preferably R7 and L2 form together with the nitrogen atom they are attached to a pyrrolidinyl or a morpholinyl group.
In one embodiment, A is CH and the compound of formula I may therefore be in racemic or optically active form. Preferably, A is CH and the compound of formula I is in optically active form. More preferably, A is CH and the compound of formula I is in (R) form.
In one embodiment, the compounds of Formula I are those of Formula II:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, A, Y, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula II are those of Formula Ha:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Y, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula II are those of Formula IIb:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula IIb are those of Formula IIc:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, Z is —NR7(CO)— and L1, L2, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
Examples of compounds according to this embodiment are those wherein one or more of L1, L2, R1, R2, R5, R6 and R7 are defined as follows:
L1 is —CH2—;
L2 is selected from —CH2—, —CH2—CH2—, —C(CH3)2— and —CH(R5)—; more preferably L2 is selected from —CH2—, —CH2—CH2— and —CH(R5)—; even more preferably L2 is —CH2—;
R1 is selected from naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 5-indolyl, 6-indolyl, 3-quinolinyl, 4-quinolinyl, 7-quinolinyl, dihydroquinolin-2-on-6-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl, styryl, 5-benzothiophenyl, 6-benzothiophenyl, 3-benzothiophenyl, 2-benzothiophenyl, 6-(1,3-benzothiazolyl and 6-benzodioxinyl, wherein said naphthalen-2-yl, phenyl, biphenyl, 4-pyrimidin-5-yl-phenyl, tetraisoquinolinyl, 3-indolyl, 5-indolyl, 6-indolyl, 3-quinolinyl, 4-quinolinyl, 7-quinolinyl, dihydroquinolin-2-on-6-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, (2H-1,2,3,4-tetrazol-5-yl)phenyl, benzyl, styryl, 5-benzothiophenyl, 6-benzothiophenyl, 3-benzothiophenyl, 2-benzothiophenyl, 6-(1,3-benzothiazolyl or 6-benzodioxinyl is optionally substituted by one or more substituents selected from C1-C4-alkyl, preferably methyl, halo, preferably bromo or chloro, more preferably bromo, C1-C4-haloalkyl, preferably C1-C4-fluroalkyl, more preferably trifluoomethyl, hydroxy-C1-C4-alkyl, preferably hydroxymethyl, acetylamino, acetylamino-C1-C4-alkyl, preferably acetylaminomethyl, and C1-C4-alkylaminocarboxyl, preferably methylaminocarboxyl;
R2 is selected from phenyl, 1,4-imidazolyl, pyrimidinyl, pyridinyl, phenyl-C1-C2-alkyl, cyclohexyl, piperidinyl, morpholinyl, 1,3-benzodioxolyl, C1-05-alkyl, t-butyloxy, phenylamino, wherein said phenyl, 1,4-imidazolyl, phenyl-C1-C2-alkyl, piperidinyl, is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, preferably methoxy, halogen, preferably fluoro and chloro, more preferably fluoro, C1-C4-haloalkyl, preferably trifluoromethyl, C1-C4-alkyl, preferably methyl, C1-C2-alkoxy-C1-C2-alkoxy, preferably methoxyethoxy, C1-C4-alkyloxycarbonyl, preferably t-butyloxycarbonyl, acetylamino, di(C1-C4-alkyl)amino-C1-C4-alkyl, preferably dimethylaminomethyl, and hydroxycarbamoyl.
R5 is selected from CH3, CH2—OH, CH2-Ph-OH and CH2-Ph-OR6;
R7 is selected from H, methyl, ethyl, isobutyl, isopentyl and (1-methylimidazol-2-yl)methyl; or
R7 and L2 form together with the nitrogen atom they are attached to a 5- or 6-membered heterocyclyl group selected from pyrrolidinyl, morpholinyl, thiazolidinyl and piperidinyl.
In one embodiment, Z is —NHSO2— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is aryl, preferably phenyl, optionally substituted by one or more substituents independently selected from halogen, preferably fluoro, and C1-C4-alkyl, preferably methyl.
In one embodiment, Z is —CH2—CH2— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is aryl, preferably phenyl.
In one embodiment, Z is —NH— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is aryl, preferably phenyl, optionally substituted by one or more substituents independently selected from halogen, preferably fluoro.
In one embodiment, Z is —NH—CH(C1-C6-alkyl)—, preferably Z is —NH—CH(C4-C6-alkyl)—, more preferably Z is —NH—CH(C5-alkyl)—, and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is C1-C4-alkyloxycarbonyl, preferably t-butyloxycarbonyl.
In one embodiment, Z is —O—CH2— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is aryl, preferably phenyl.
In one embodiment, Z is —C(O)NH— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2— or —CH2—CH2—; R2 is aryl, preferably phenyl, optionally substituted by one or more substituents independently selected from halogen, preferably fluoro.
In one embodiment, Z is —O— and L1 is —CH2—; R1 is aryl, preferably naphthalen-2-yl; L2 is —CH2—; R2 is aryl, preferably phenyl, optionally substituted by acetylamino.
In one embodiment, preferred compounds of Formula IIc are those of Formula IId:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L2, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula II are those of Formula lie:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula IIe are those wherein:
L1 is —CH2—;
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthalen-2-yl;
L2 is —CH2—;
R2 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, and C1-C4-haloalkyl; preferably R2 is phenyl substituted by one or more substituents selected from halogen, preferably fluor.
In one embodiment, the compounds of Formula I are those of Formula III:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, A, Y, Z, R1, R2, R3, R4, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula III are those of Formula Ma:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Y, Z, R1, R2, R3, R4, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula III are those of Formula IIIb:
and pharmaceutically acceptable salts and solvates thereof,
wherein
R1, R2, R3, R4 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula Ma are those wherein:
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthalen-2-yl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl and C1-C6-alkyl, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; preferably R2 is selected from aryl, arylalkyl, cycloalkyl and C1-C6-alkyl, wherein said aryl or arylalkyl is optionally substituted by one substituent selected from C1-C4-alkoxy and halogen; more preferably R2 is selected from phenyl, cyclohexyl, phenylethyl, naphthalen-2-yl-CH(CH3)— and methyl, wherein said phenyl or naphthalen-2-yl-CH(CH3)— is optionally substituted by one substituent selected from C1-C4-alkoxy, preferably methoxy, and halogen, preferably fluor.
R3 is selected from H and C(O)OR4; preferably R3 is H;
R4 is C1-C4-alkyl; preferably R3 is ethyl;
R7 is selected from H and C1-C6-alkyl; preferably R7 is H.
In one embodiment, the compounds of Formula I are those of Formula IV:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, A, Y, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula IV are those of Formula IVa:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Y, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula IVa are those wherein:
L1 is —CH2—;
L2 is —CH2—;
R7 is selected from H and C1-C6-alkyl; preferably R7 is H;
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthalen-2-yl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl and C1-C6-alkyl, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; preferably R2 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; more preferably R2 is aryl, optionally substituted by one substituent selected from halogen; still more preferably R2 is phenyl, optionally substituted by one halogen, preferably fluor; even more preferably R2 is phenyl substituted by one fluor.
In one embodiment, preferred compounds of Formula IV are those of Formula IVb:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, R1 and R2 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula IVb are those wherein:
L1 is inexistent or —CH2—; preferably L1 is inexistent.
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthalen-2-yl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl and C1-C6-alkyl, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; preferably R2 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; more preferably R2 is aryl, optionally substituted by one substituent selected from halogen; still more preferably R2 is phenyl, optionally substituted by one halogen, preferably fluor; even more preferably R2 is phenyl substituted by one fluor.
In one embodiment, the compounds of Formula I are those of Formula V:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, A, Y, Z, R1, R2, R5, R6 and R7 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, preferred compounds of Formula V are those of Formula Va:
and pharmaceutically acceptable salts and solvates thereof,
wherein
L1, L2, Y, Z, R1, R2, R5 and R6 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula Va are those wherein:
R6 is selected from H and C1-C6-alkyl; preferably R6 is H;
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthyl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl and C1-C6-alkyl, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; preferably R2 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; more preferably R2 is aryl, optionally substituted by one substituent selected from halogen; still more preferably R2 is phenyl, optionally substituted by one halogen, preferably fluor; even more preferably R2 is phenyl substituted by one fluor.
In one embodiment, preferred compounds of Formula V are those of Formula Vb:
and pharmaceutically acceptable salts and solvates thereof,
wherein
R1 and R2 are as defined above with respect to Formula I and any of its embodiments.
In one embodiment, the compounds of Formula Vb are those wherein:
R1 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen and C1-C4-haloalkyl; preferably R1 is naphthyl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl and C1-C6-alkyl, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; preferably R2 is aryl optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl and C1-C4-alkyl; more preferably R2 is aryl, optionally substituted by one substituent selected from halogen; still more preferably R2 is phenyl, optionally substituted by one halogen, preferably fluor; even more preferably R2 is phenyl substituted by one fluor.
Particularly preferred compounds of the invention are those listed in Table 1 hereafter:
The compounds of the invention can be prepared by different ways with reactions known by the person skilled in the art. Reaction schemes as described in the example section illustrate by way of example different possible approaches.
The compounds of Formula I are indeed capable of sensitizing cancer cells to various anticancer drugs. They further have the advantage of increasing the cytoxicity of cytotoxic agents, preferably proteasome inhibitors or HDAC inhibitors. The invention thus also provides the use of the compounds of Formula I or pharmaceutically acceptable salts, or solvates thereof as sensitizers for chemotherapy of malignant tumors.
Accordingly, in a particularly preferred embodiment, the invention relates to the use of compounds of Formula I and subformulae, in particular those of Table 1 above, or pharmaceutically acceptable salts and solvates thereof, for the treatment and/or prevention of cancer, particularly as sensitizers for chemotherapy of malignant tumors.
Unexpectedly, the inventors have discovered that the compounds of Formula I, including the 18 compounds listed in the proviso above, may be used as sensitizers for chemotherapy of malignant tumors.
As disclosed in the prior art, the 18 compounds listed in the proviso above may be of particular interest in animal models of chronic or degenerative diseases such as type-2 diabetes and Alzheimer's disease (R. Deprez-Poulain et al., Nature Communications, 2015, 6, article number:8250). However, the inventors have found that compounds of Formula I are capable of improving the effect of various cytotoxic agents for chemotherapy, and these compounds are therefore part of the use according to the invention.
The compounds for use according to the invention therefore include compounds of Formula I and subformulae II, IIa, IIb, IIc, IId, IIe, III, IIIa, IIIb, IV, IVa, IVb, V, Va and Vb as defined above, in particular compounds of Table 1 above.
The compounds of the invention are able to sensitize cancer cells to various anticancer agents in various cancers, and therefore to increase the cytotoxicity of those anticancer agents.
The compounds of Formula I, or any of its subformulae as defined above, can thus be used as sensitizers for chemotherapy of malignant tumors, aiming at improving the chemotherapy effect of the cancer treatment, preventing tolerance and decreasing toxicity and adverse effects.
The invention thus relates to a compound of Formula I:
or a pharmaceutically acceptable salt or solvate thereof for use in treating and/or preventing cancer,
wherein
L1 is inexistent or is selected from —CH2— and —CH(OH)—;
R1 is selected from aryl, heteroaryl, arylalkyl and arylalkenyl wherein said aryl, heteroaryl, arylalkyl or arylalkenyl is optionally substituted by one or more substituents independently selected from C1-C4-alkyl, halogen, C1-C4-haloalkyl, hydroxy-C1-C4-alkyl, acetylamino, acetylamino-C1-C4-alkyl and C1-C4-alkylaminocarboxyl;
R2 is selected from aryl, heteroaryl, arylalkyl, cycloalkyl, heterocycloalkyl, C1-C6-alkyl, C1-C4-alkyloxycarbonyl, C1-C4-alkoxy, arylamino, wherein said aryl, arylalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from C1-C4-alkoxy, halogen, C1-C4-haloalkyl, C1-C4-alkyl, C1-C2-alkoxy-C1-C2-alkoxy, C1-C4-alkyloxycarbonyl, acetylamino, di(C1-C4-alkyl)amino-C1-C4-alkyl and hydroxycarbamoyl;
X is selected from
R3 is selected from H and C(O)OR4;
R4 is C1-C4-alkyl;
L2 is linear or branched C1-C6-alkyl, optionally substituted by a group R5;
R5 is selected from linear or branched C1-C4-alkyl, CH2—OH, CHOH—CH3, CH2—C(O)NH2, CH2—CH2—C(O)NH2, CH2—COOH, CH2—CH2—COOH, (CH2)4—NH2, (CH2)4—NH—C(NH2+)—NH2, CH2—Imidazolyl, CH2—Indolyl, CH2—SH, CH2—CH2—S—CH3, CH2-Ph, CH2-Ph-OH, CH2—OR6, CH2—COOR6, CH2—CH2—COOR6, CH2—SR6 and CH2-Ph-OR6;
R6 is selected from Me, Bn, Ph and Ac;
Z is selected from —NR7(CO)—, —NHSO2—, —CH2—CH2—, —NH—, —NH—CH(C1-C6-alkyl)—, —O—CH2—, —C(O)NH— and —O—; and
R7 is selected from H, linear or branched C1-C6-alkyl and (1-methylimidazol-2-yl)-C1-C2-alkyl, or
R7 and L2 form together with the nitrogen atom they are attached to a 5- or 6-membered heterocyclyl group.
In one embodiment, the invention relates to a compound of any of the subformulae of Formula I as defined above, in particular a compound of Table 1 above, or a pharmaceutically acceptable salt or solvate thereof, for use in treating and/or preventing cancer, particularly as sensitizers for chemotherapy of malignant tumors.
Anticancer agents, or cytotoxic agents, within the meaning of the present invention include, but are not limited to, proteasome inhibitors, HDAC inhibitors and protein glycosylation inhibitors. Preferred proteasome inhibitors are epoxomycin, carfilzomib, bortezomib, oprozomib, ixazomib and nelfinavir. More preferred proteasome inhibitors are epoxomycin, carfilzomib and nelfinavir. Preferred HDAC inhibitors are panobinostat, quisinostat and N-[4-[3-[[[7-(hydroxyamino)-7-oxoheptyl]amino]carbonyl]-5-isoxazolyl]phenyl]-1,1-dimethylethyl ester, carbamic acid (CAS#: 1045792-66-2). A more preferred HDAC inhibitor is panobinostat. A preferred protein glycosylation inhibitor is tunicamycin.
Thus, in one embodiment, there is provided a compound of Formula I, or any of its subformulae as defined above, for use as sensitizer for chemotherapy of malignant tumors involving a proteasome inhibitor, a HDAC inhibitor or a protein glycosylation inhibitor.
The compounds of the invention are therefore useful in the prevention and/or treatment of cancers, and particularly cancers that may be treated by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors.
The invention thus also relates to a compound of the invention or a pharmaceutically acceptable salt or solvate thereof for use in the treatment and/or prevention of cancer, in particular cancers that may be treated by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors. Or in other terms, the invention also relates to a method of treating and/or preventing cancer, in particular cancers that may be treated by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors, comprising the administration of a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt or solvate, to a patient in need thereof. Preferably the patient is a warm-blooded animal, more preferably a human.
Cancers that may be treated by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors, within the meaning of the present invention include, but are not limited to, multiple myeloma, cervical cancer and breast cancer.
The invention further provides the use of a compound of the invention or a pharmaceutically acceptable salt or solvates thereof for the manufacture of a medicament for use in treating and/or preventing cancer, in particular cancers that may be treated by cytotoxic agents, preferably by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors. Preferably the patient is a warm-blooded animal, more preferably a human. The cancers that may be treated by proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors are preferably those defined above.
According to a further feature of the present invention, there is provided a compound of Formula I or a pharmaceutically acceptable salt or solvate for use in sensitizing cancer cells to anticancer drugs, preferably proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors, in a patient in need of such treatment, comprising administering to said patient an effective amount of a compound of the invention, or a pharmaceutically acceptable salt or solvate thereof. In other terms, the invention also provides a method for sensitizing cancer cells to anticancer drugs, preferably proteasome inhibitors, HDAC inhibitors or protein glycosylation inhibitors, in a patient in need of such treatment, which comprises administering to said patient an effective amount of a compound of the invention, or a pharmaceutically acceptable salt or solvate thereof. Preferably, the patient is a warm blooded animal, and even more preferably a human.
According to one embodiment, the compounds of the invention, their pharmaceutical acceptable salts or solvates may be administered as part of a combination therapy. Thus, are included within the scope of the present invention embodiments comprising co-administration of, and compositions and medicaments which contain, in addition to a compound of the present invention, a pharmaceutically acceptable salt or solvate thereof as active ingredient, additional therapeutic agents and/or active ingredients. Such multiple drug regimens, often referred to as combination therapy, may be used in the treatment and/or prevention of any cancer, particularly those defined above.
Thus, the methods of treatment and pharmaceutical compositions of the present invention may employ the compounds of the invention or their pharmaceutical acceptable salts or solvates thereof in the form of monotherapy, but said methods and compositions may also be used in the form of multiple therapy in which one or more compounds of the invention or their pharmaceutically acceptable salts or solvates are co-administered in combination with one or more other therapeutic agents. Such additional therapeutic agents include, but are not limited to, proteasome inhibitors, HDAC inhibitors and protein glycosylation inhibitors. Preferred proteasome inhibitors are epoxomycin, carfilzomib, bortezomib, oprozomib, ixazomib and nelfinavir. More preferred proteasome inhibitors are epoxomycin, carfilzomib and nelfinavir. Preferred HDAC inhibitors are panobinostat, quisinostat and N-[4-[3-[[[7-(hydroxyamino)-7-oxoheptyl]amino]carbonyl]-5-isoxazolyl]phenyl]-1,1-dimethylethyl ester, carbamic acid (CAS#: 1045792-66-2). A more preferred HDAC inhibitor is panobinostat. A preferred protein glycosylation inhibitor is tunicamycin.
In one embodiment, the methods of treatment and pharmaceutical compositions of the present invention may employ the compounds of Formula I, or any of its subformulae as defined above, or their pharmaceutical acceptable salts or solvates thereof, in combination with radiation therapy. According to this embodiment, the compounds of the invention, their pharmaceutical acceptable salts or solvates may be administered in combination with radiation therapy. Thus, there is provided a compound of Formula I, or any of its subformulae as defined above, for use in the treatment and/or prevention of cancers as defined above in combination with radiation therapy. Such radiation therapies include, but are not limited to, external beam radiation therapy, brachytherapy and systemic radioisotope therapy.
The invention also provides pharmaceutical compositions comprising a compound of the invention or a pharmaceutically acceptable salt or solvate thereof and at least one pharmaceutically acceptable carrier, diluent, excipient and/or adjuvant. As indicated above, the invention also covers pharmaceutical compositions which contain, in addition to a compound of the present invention, a pharmaceutically acceptable salt or solvate thereof as active ingredient, additional therapeutic agents and/or active ingredients.
Another object of this invention is a medicament comprising at least one compound of the invention, or a pharmaceutically acceptable salt or solvate thereof, as active ingredient.
Generally, for pharmaceutical use, the compounds of the invention may be formulated as a pharmaceutical preparation comprising at least one compound of the invention and at least one pharmaceutically acceptable carrier, diluent, excipient and/or adjuvant, and optionally one or more further pharmaceutically active compounds.
By means of non-limiting examples, such a formulation may be in a form suitable for oral administration, for parenteral administration (such as by intravenous, intramuscular or subcutaneous injection or intravenous infusion), for topical administration (including ocular), cerebral administration, for administration by inhalation, by a skin patch, by an implant, by a suppository, etc. Such suitable administration forms—which may be solid, semi-solid or liquid, depending on the manner of administration—as well as methods and carriers, diluents and excipients for use in the preparation thereof, will be clear to the skilled person; reference is made to the latest edition of Remington's Pharmaceutical Sciences.
The definitions and explanations below are for the terms as used throughout the entire application, including both the specification and the claims.
Unless otherwise stated any reference to compounds of the invention herein, means the compounds as such as well as their pharmaceutically acceptable salts and solvates.
When describing the compounds of the invention, the terms used are to be construed in accordance with the following definitions, unless indicated otherwise.
The term “unsubstituted” as used herein means that a radical, a group or a residue carries no substituents. The term “substituted” means that a radical, a group or a residue carries one or more substituents.
The term “halo” or “halogen” refers to the atoms of the group 17 of the periodic table (halogens) and includes in particular fluorine, chlorine, bromine and iodine atom. Preferred halo groups are fluoro and chloro, fluoro being particularly preferred.
The term “alkyl” by itself or as part of another substituent refers to a hydrocarbyl radical of Formula CnH2n+1 wherein n is a number greater than or equal to 1. Alkyl groups may be linear or branched. Examples of alkyl groups include but are not limited to methyl (abbreviation: Me), ethyl (abbreviation: Et), n-propyl (abbreviation: n-Pr), isopropyl (abbreviation: i-Pr), n-butyl (abbreviation: n-Bu), isobutyl (abbreviation: i-Bu), tert-butyl (abbreviation: t-Bu), sec-butyl (abbreviation: s-Bu), n-pentyl, isopentyl, hexyl and isohexyl.
The term “haloalkyl” alone or in combination, refers to an alkyl radical having the meaning as defined above wherein one or more hydrogens are replaced with a halogen as defined above. Non-limiting examples of such haloalkyl radicals include chloromethyl, 1-bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1,1,1-trifluoroethyl and the like.
The term “cycloalkyl” as used herein is a monovalent, saturated, or unsaturated monocyclic or bicyclic hydrocarbyl group. Cycloalkyl groups may comprise 3 or more carbon atoms in the ring and generally, according to this invention comprise from 3 to 10, more preferably from 3 to 8 carbon atoms still more preferably from 3 to 6 carbon atoms. Examples of cycloalkyl groups include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
The term “heteroatom” as used herein refers to any atom that is not carbon or hydrogen. Non-limiting examples of such heteroatoms include nitrogen, oxygen, sulfur, and phosphorus. Preferred heteroatoms according to the invention are nitrogen, oxygen and sulfur.
The terms “heterocyclyl”, “heterocycloalkyl” or “heterocyclo” as used herein by itself or as part of another group refer to non-aromatic, fully saturated or partially unsaturated cyclic groups (for example, 3 to 7 member monocyclic, 7 to 11 member bicyclic, or containing a total of 3 to 10 ring atoms) which have at least one heteroatom in at least one carbon atom-containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1, 2, 3 or 4 heteroatoms selected from nitrogen, oxygen and/or sulfur atoms, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom of the ring or ring system, where valence allows. Examples of heterocyclyl groups include but are not limited to aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, piperazinyl, morpholinyl. Particular heterocyclyl groups according to the invention are pyrrolidinyl, morpholinyl, piperidinyl and thiazolidinyl. Preferred heterocyclyl groups according to the invention are pyrrolidinyl and morpholinyl.
The term “aryl” as used herein refers to a polyunsaturated, aromatic hydrocarbyl group having a single ring (i.e. phenyl) or multiple aromatic rings fused together (e.g. naphthyl), typically containing 5 to 12 atoms; preferably 6 to 10, wherein at least one ring is aromatic. Examples of aryl groups include but are not limited to phenyl (abbreviation: Ph), biphenyl, 1-naphthyl (or naphthalene-1-yl), 2-naphthyl (or naphthalene-2-yl), anthracenyl, indanyl, indenyl, 1,2,3,4-tetrahydronaphthyl. Preferred aryl groups according to the invention are phenyl, biphenyl and 2-naphthyl.
The term “heteroaryl” as used herein by itself or as part of another group refers but is not limited to 5 to 12 carbon-atom aromatic rings or ring systems containing 1 to 2 rings which are fused together, typically containing 5 to 6 atoms; at least one of which is aromatic, in which one or more carbon atoms in one or more of these rings is replaced by oxygen, nitrogen and/or sulfur atoms where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quaternized. Examples of heteroaryl groups include but are not limited to pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, furanyl, benzofuranyl, pyrrolyl, indolyl, thiophenyl, benzothiophenyl, imidazolyl, benzimidazolyl, pyrazolyl, indazolyl, oxazolyl, benzoxazolyl, isoxazolyl, benzisoxazolyl, thiazolyl, and benzothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, dioxazolyl, dithiazolyl and tetrazolyl. Particular heteroaryl groups according to the invention are triazolyl, oxadiazolyl, pyrazolyl, imidazolyl, tetrazolyl, quinolinyl, indolyl, benzothiophenyl, dihydroquinolinonyl, benzodioxolyl and benzodioxinyl. Preferred heteroaryl groups according to the invention are triazolyl, oxadiazolyl, pyrazolyl, imidazolyl, tetrazolyl, quinolinyl and indolyl.
The term “arylalkyl” or “aralkyl” as used herein refers to a group-alkyl-aryl, wherein alkyl and aryl are as herein defined. Examples of arylalkyl groups include but are not limited to benzyl (abbreviation: Bn).
The term “arylalkenyl” as used herein refers to a group -alkenyl-aryl, wherein alkenyl and aryl are as herein defined. Examples of arylalkenyl groups include but are not limited to styryl.
The term “alkoxy” as used herein refers to a group —O-alkyl, wherein alkyl is as herein defined. Examples of alkoxy groups include but are not limited to methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butyloxy, t-butyloxy, sec-butyloxy and n-pentyloxy.
The term “acetyl” or its abbreviation “Ac” as used herein refers to a group —C(O)CH3.
The compounds of the invention containing a basic functional group may be in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts of the compounds of the invention containing one or more basic functional groups include in particular the acid addition salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include the acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, cinnamate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate salts.
Pharmaceutically acceptable salts of compounds of Formula I and subformulae may for example be prepared as follows:
(i) reacting the compound of Formula I or any of its subformulae with the desired acid; or
(ii) converting one salt of the compound of Formula I or any of its subformulae to another by reaction with an appropriate acid or by means of a suitable ion exchange column.
All these reactions are typically carried out in solution. The salt, may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the salt may vary from completely ionized to almost non-ionized.
The term “solvate” is used herein to describe a molecular complex comprising the compound of the invention and one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term “hydrate” is employed when said solvent is water.
The compounds of the invention include compounds of the invention as hereinbefore defined, including all polymorphs and crystal habits thereof, prodrugs and isomers thereof (including optical, geometric and tautomeric isomers) and isotopically-labeled compounds of the invention.
In addition, although generally, with respect to the salts of the compounds of the invention, pharmaceutically acceptable salts are preferred, it should be noted that the invention in its broadest sense also includes non-pharmaceutically acceptable salts, which may for example be used in the isolation and/or purification of the compounds of the invention. For example, salts formed with optically active acids or bases may be used to form diastereoisomeric salts that can facilitate the separation of optically active isomers of the compounds of the invention.
The term “patient” refers to a warm-blooded animal, more preferably a human, who/which is awaiting or receiving medical care or is or will be the object of a medical procedure.
The term “human” refers to subjects of both genders and at any stage of development (i.e. neonate, infant, juvenile, adolescent, adult). In one embodiment, the human is an adolescent or adult, preferably an adult.
The terms “treat”, “treating” and “treatment”, as used herein, are meant to include alleviating or abrogating a condition or disease and/or its attendant symptoms.
The terms “prevent”, “preventing” and “prevention”, as used herein, refer to a method of delaying or precluding the onset of a condition or disease and/or its attendant symptoms, barring a patient from acquiring a condition or disease, or reducing a patient's risk of acquiring a condition or disease.
The term “therapeutically effective amount” (or more simply an “effective amount”) as used herein means the amount of active agent or active ingredient which is sufficient to achieve the desired therapeutic or prophylactic effect in the individual to which it is administered.
The term “administration”, or a variant thereof (e.g., “administering”), means providing the active agent or active ingredient, alone or as part of a pharmaceutically acceptable composition, to the patient in whom/which the condition, symptom, or disease is to be treated or prevented.
By “pharmaceutically acceptable” is meant that the ingredients of a pharmaceutical composition are compatible with each other and not deleterious to the patient thereof.
The term “agonist” as used herein means a ligand that activates an intracellular response when it binds to a receptor.
The term “pharmaceutical vehicle” as used herein means a carrier or inert medium used as solvent or diluent in which the pharmaceutically active agent is formulated and/or administered. Non-limiting examples of pharmaceutical vehicles include creams, gels, lotions, solutions, and liposomes.
The present invention will be better understood with reference to the following examples and FIGURES. These examples are intended to representative of specific embodiments of the invention, and are not intended as limiting the scope of the invention.
All reagents, solvents and starting materials were purchased from commercial suppliers and used without further purification.
Melting points were determined using a Büchi B-540 melting point apparatus and are uncorrected.
1H NMR spectra were recorded on a Brucker Avance 300 MHz spectrometer with methanol-d4, CDCl3, DMSO-d6 or acetone-d6 as the solvent. 13C NMR spectra are recorded at 100 MHz. All coupling constants are measured in hertz (Hz) and the chemical shifts (δ) are quoted in parts per million (ppm).
Liquid chromatography mass spectroscopy analyses (LC-MS) were performed using LCMS-MS triple-quadrupole system (Waters) with a C18 TSK-GEL Super ODS (2 μm particle size column, 50×4.6 mm). LCMS gradient starting from 98% H2O/0.1% formic acid and reaching 2% H2O/98% MeOH within 5 min (method A) at a flow rate of 2 mL/min or starting from 100% H2O/0.1% formic acid and reaching 5% H2O/95% MeOH within 10 min (method B) at a flow rate of 1 mL/min was used. Purity (%) was determined by Reversed Phase HPLC, using UV detection (215 nM).
High resolution mass spectroscopy (HRMS) were carried out on an Waters LCT Premier XE (TOF), ESI ionization mode, with a Waters XBridge C18 (150×4.6 mm, 3.5 μm particle size). LCMS gradient starting from 98% ammonium formate buffer 5 mM (pH 9.2) and reaching 95% CH3CN/5% ammonium formate buffer 5 mM (pH 9.2) within 15 min at a flow rate of 1 mL/min was used.
Solvents, reagents and starting materials were purchased from well known chemical suppliers such as for example Sigma Aldrich, Acros Organics, Fluorochem, Eurisotop, VWR International, Sopachem and Polymer labs and the following abbreviations are used:
AUC: Area under curve,
CC50: 50% cytotoxic concentration,
EDCI: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide,
EDTA: Ethylenediaminetetraacetic acid,
eq or equiv: Equivalent,
KCN: Potassium cyanide,
HRMS: High resolution mass spectrometry,
LC: Liquid chromatography,
LCMS: Liquid chromatography-mass spectrometry,
Mp: Melting point,
MS: Mass spectrometry,
MW: Molecular weight,
NMR: Nuclear magnetic resonance,
PE: Petroleum ether,
RT or rt: Room temperature,
THF: tetrahydrofurane,
TLC: Thin layer chromatography,
TFA: Trifluoroacetic acid,
tR: Retention time,
CFZ: carfilzomib,
TUN: tunicamycin,
BTZ: bortezomib,
OZ: oprozomib,
PAN: panobinostat,
NLF: nelfinavir,
EPX: epoxomycin.
1. General Protocols
(a) General Procedure A for Azide Synthesis:
Imidazole-1-sulfonyl azide hydrochloride (1.2 equiv) was added to the β-amino ester (1 equiv), K2CO3 (1 equiv) and CuSO4.5H2O (0.01 equiv) in methanol and the mixture was stirred at room temperature overnight. The mixture was concentrated, diluted with H2O, acidified with concentrated HCl and extracted with ethyl acetate (3 times). The combined organic layers were washed with brine, dried (MgSO4), filtered and concentrated to give the azide which was purified if needed.
(b) General Procedure A′ for Azide Synthesis:
The starting β-amino ester (1.0 equiv) was dissolved in dry MeOH under argon, and cooled down to 0° C. before ZnCl2 (0.06 equiv), potassium carbonate (1.0 equiv) and DIPEA (1.0 equiv) were added. In a separate flask, cooled DIPEA (1.15 equiv) was added to a solution of N-diazoimidazole-1-sulfonamide (as a bisulfate salt, 1.05 equiv) in MeOH at 0° C. The resulting solution was immediately added dropwise to the β-amino ester solution, and the resulting mixture was further stirred overnight in the melting ice bath. The reaction was cooled back down to 0° C., diluted with water and carefully acidified to pH=2 with 1M KHSO4 and extracted with ethyl acetate. Organic layers were mixed and concentrated to dryness in vacuo. Purification over silica gel column was performed if needed.
(c) General Procedure A″ for Azide Synthesis:
The starting β-amino ester (1.0 equiv) was dissolved in dry MeOH under argon, and cooled down to 0° C. before ZnCl2 (0.06 equiv), potassium carbonate (1.0 equiv) and DIPEA (1.0 equiv) were added. In a separate flask, cooled DIPEA (1.15 equiv) was added to a solution of N-diazoimidazole-1-sulfonamide (as a bisulfate salt, 1.05 equiv) in MeOH at 0° C. The resulting solution was immediately added dropwise to the β-amino ester solution, and the resulting mixture was further stirred overnight in the melting ice bath. The reaction was cooled back down to 0° C., diluted with water and extracted with ethyl acetate. Organic layers were mixed and concentrated to dryness in vacuo. Purification over silica gel column was performed if needed.
(d) General Procedure B for 1,4-disubstituted 1,2,3-triazole Synthesis:
The azide (1 equiv) and the alkyne (1 equiv) were dissolved separately in DMSO (100-150 (IL) then added to a mixture of tBuOH/water (1:1 (v/v)) or directly solubilised in a mixture of DMF/water (1:1 (v/v)). CuSO4:5H2O (0.1 equiv) and sodium ascorbate (1 equiv) were added. After 16 h of stirring at room temperature, the media was filtered. In most cases, concentration of the filtrate, followed by precipitation in water, filtration and washing with ethyl acetate gave a pure product. If necessary, the final triazole was purified.
(e) General Procedure C for 1,4-disubstituted 1,2,3-triazole Synthesis:
Azide (1.0 equiv) and alkyne (1.2 equiv) were added in a 1:1 (v/v) mixture of water and DMF or dioxane or a 2:1:1 (v/v) mixture of CH3CN, dioxane and water. Sodium ascorbate (0.3 equiv) was added, followed by copper(II) sulfate pentahydrate (0.06 equiv). The mixture was stirred overnight. Solvents were evaporated under reduced pressure and the residue was diluted with water and extracted three times with EtOAc. The combined organic layers were dried with MgSO4, filtered and concentrated under reduced pressure to give the triazole which was purified if needed.
(f) General Procedure C′ for 1,4-disubstituted 1,2,3-triazole Synthesis:
Azide (1.0 equiv) and alkyne (1.05 equiv) were dissolved in DMF or dioxane. A solution of copper(II) sulfate pentahydrate (0.1 equiv) in distilled water was added, quickly followed by sodium ascorbate (0.5 equiv). The mixture was stirred overnight before it was diluted with water and extracted three times with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give the triazole which was purified if needed.
(g) General Procedure D for Hydroxamic Acid Synthesis:
The carboxylic acid (1.0 equiv) was dissolved in DMF. EDCI (1.2 equiv), HOBt (1.5 equiv), N-methylmorpholine (6.0 equiv) were added. The mixture was stirred at room temperature for 5 min, and O-tritylhydroxylamine (1.2 equiv) was added. The mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2Cl2 and washed three times with a 5% NaHCO3 (aq) solution and once with water. The organic layer was dried with MgSO4, filtered and concentrated under reduced. The crude product was purified by flash chromatography on silica gel (CH2Cl2/MeOH). The obtained O-trityl hydroxamate intermediate was dissolved in TFA 5%/CH2Cl2 and triisopropylsilane was added. The mixture was stirred at room temperature for 30 min. Solvents were removed under reduced pressure and the residue was triturated with diethyl ether and petroleum ether to give a residue which was purified if needed to afford the hydroxamic acid.
(h) General Procedure E for Hydroxamic Acid Synthesis:
The azido ester (1 equiv) was dissolved in MeOH. HONH2.HCl (7.2 equiv) was dissolved in MeOH. KOH (11.4 equiv) was dissolved in MeOH. The KOH solution was poured into the HONH2.HCl solution, and the resulting mixture was cooled to 0° C. for 1 h. The KOH/HONH2 solution was then filtered into the solution of the ester, and the reaction mixture was stirred at room temperature until completion. After removal of the solvent, the mixture was dissolved in ethyl acetate and washed successively with a 1N solution of HCl and brine. Removal of the solvent gave a brown oil which was purified if needed.
(i) General Procedure F for Hydroxamic Acid Synthesis:
A solution of hydroxylamine in water (50% in water, 1 equiv) and MeOH was dried with an excess of Na2SO4 and filtered. The filtrate was added to a flask containing the ester (1 equiv) and KCN (1 equiv). The mixture was stirred until completion of the reaction. The reaction mixture was diluted with CH3CN. The unsoluble solids were filtered. The filtrate was evaporated to dryness and purified if needed to give the desired hydroxamic acid.
(j) General Procedure G for Hydroxamic Acid Synthesis:
To a solution of the ester (1 equiv) in a 1:1 (v/v) mixture of MeOH and NH2OH (50% in water) was added KCN (0.1 equiv). The mixture was stirred at room temperature for 16 h. The mixture was diluted with water and EtOAc. The aqueous phase was extracted twice with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to afford the desired compounds, which was purified if needed.
(k) General Procedure G′ for Hydroxamic Acid Synthesis:
To a solution of the ester (1 equiv) in a 1:1 (v/v) mixture of MeOH and NH2OH (50% in water) was added KCN (0.1-0.5 equiv). The mixture was stirred at room temperature for 4-16 h. All solvents were evaporated in vacuo before purification.
(l) General Procedure H for Carboxylic Acid Methylation:
To a stirred solution of the corresponding amino acid (1 equiv) in methanol (107 equiv) was added dropwise thionyl chloride (15 equiv) at 0° C. After stirring at room temperature overnight, the solvent was evaporated and the crude material precipitated in ether to give the methyl ester.
(m) General Procedure I for Amine Acylation:
The amine (1 equiv) was dissolved in dioxane and an aqueous saturated solution of K2CO3. The biphasic mixture was cooled to 0° C. A solution of acyl chloride (1 equiv) in dioxane was slowly added to the amino acid solution and the reaction was allowed to slowly warm to room temperature. After 3 hours, the reaction was dissolved in water and extracted with diethyl ether (twice). The resulting aqueous layer was acidified to pH 1 with 3 M HCl (aq) and extracted with ethyl acetate (3 times). The combined organic layers were dried with MgSO4, filtered and concentrated under reduced pressure to give the desired compound after purification if needed.
(n) General Procedure J for Negishi Coupling:
Zinc dust (3 equiv) was added to a dry round bottom 100 mL flask under argon. Dry DMF was added via a syringe followed by a catalytic amount of iodine (0.15 equiv). Compound 123 (1 equiv) was added immediately followed by a catalytic amount of iodine (0.15 equiv). The solution was stirred at room temperature and gave a noticeable exotherm. When the solution had cooled Pd2dba3 (0.025 equiv), SPhos (0.05 equiv) and the aromatic bromide or iodide (1.3 equiv) were added to the flask and left to stir at room temperature overnight, under argon. The crude reaction mixture was purified by flash chromatography to afford the desired compound.
(o) General Procedure K for Negishi Coupling:
Zinc dust (6 equiv) was added to a dry round-bottom flask under argon. Dry DMF and TMSCl (1.3 equiv) were added via a syringe. The reaction mixture was sonicated then stirred at room temperature for 5 min. The zinc was allowed to settle and the supernatant solution was removed via a syringe, followed by drying of the zinc under vacuum using heat from a heat-gun. Compound 123 (1 equiv) was dissolved in DMF (1.42 mL) and flushed under argon then transferred to the zinc via a syringe. The solution was stirred at room temperature 5 min then Pd2(dba)3 (0.03 equiv), SPhos (0.06 equiv) and the aromatic bromide or iodide (1.3 equiv) were added to the flask. The reaction mixture was stirred at room temperature overnight. The DMF was evaporated under reduced pressure. The crude reaction mixture was purified by flash chromatography to afford the desired compound.
(p) General Procedure L for Negishi Coupling:
Zinc dust (3-5 equiv) was added to a dry sealable tube under argon. A solution of iodine (0.25 equiv) in dry DMF was added. After the brown-purple colour disappeared, TMSCl (0.25 equiv) was added. After 2 min, stirring was stopped and all insolubles were allowed to settle for 10 min. The supernatant was removed via a syringe, followed by drying of the zinc under vacuum using heat from a heat-gun. The tube was cooled down to 0° C. before a solution of compound 123 (1 equiv) and a crystal of iodine in dry DMF was added dropwise. After 15 min, Pd2(dba)3 (0.025 equiv), SPhos (0.05 equiv) and the aromatic bromide or iodide (1.2 equiv) were added and the resulting mixture was stirred at room temperature overnight. The resulting mixture was filtered, diluted with water and extracted thrice with EtOAc. The collected organic phases were dried over MgSO4, filtered and concentrated under reduced pressure to give the desired compound. Purification was performed if needed.
(q) General Procedure M for Negishi Coupling:
Zinc dust (3-5 equiv) was added to a dry sealable tube under argon. A solution of iodine (0.25 equiv) in dry DMF was added. After the brown-purple colour disappeared, TMSCl (0.25 equiv) was added. After 2 min, stirring was stopped and all insolubles were allowed to settle for 10 min. The supernatant was removed via a syringe, followed by drying of the zinc under vacuum using heat from a heat-gun. A solution of iodine (1-3 crystals) in dry DMF was added to the zinc, followed by Pd2(dba)3 (0.025 equiv), SPhos (0.05 equiv) and the aromatic bromide or iodide (1.2 equiv) and the resulting mixture was cooled down to 0° C. A solution of the compound 123 (1 equiv) in dry DMF was added dropwise over 5 h at 0° C. The resulting mixture was stirred at room temperature overnight. The resulting mixture was filtered, diluted with water and extracted thrice with EtOAc. The collected organic phases were dried over MgSO4, filtered and concentrated under reduced pressure to give the desired compound. Purification was performed if needed.
(r) General Procedure N for Amine Deprotection:
N-Boc protected compound (1 equiv) was dissolved in CH2Cl2 and TFA. The mixture was stirred at room temperature until completion. Solvents were removed under reduced pressure to give the desired compound. Purification was performed if needed.
(s) General Procedure O for Amine Deprotection:
To a solution of N-Boc protected compound (1 equiv) in MeOH at 0° C. was added a solution of 4N HCl in dioxane. The resulting mixture was stirred overnight at room temperature. Solvents were removed under reduced pressure. The residue was partitioned between aq. 0.5N HCl and CH2Cl2. The aqueous phase was basified by careful addition of solid NaHCO3. The basic aqueous layer was extracted thrice with CH2Cl2. The collected organic phases were dried over MgSO4, filtered and concentrated under reduced pressure to give the desired compound. Purification was performed if needed.
(t) General Procedure O′ for Amine Deprotection:
The starting Boc-protected β-amino-ester was put in solution in DCM and 2,6-lutidine (10 equiv). TMSOTf (5 equiv) was added dropwise and the mixture was stirred overnight at room temperature. The reaction was quenched by addition of MeOH. It was washed with 5% NaHCO3 and the aqueous phase was further extracted with DCM twice. Organic layers were combined and concentrated to dryness in vacuo. The product was purified through silica gel column when needed.
(u) General Procedure P for Saponification:
To a solution the ester (1 equiv) in THF/EtOH (2:1 (v/v)) was added 2 M NaOH. The mixture was stirred at room temperature overnight. 1 N HCl was added and the mixture was diluted with water and CH2Cl2. The aqueous layer was extracted twice with CH2Cl2. The collected organic phases were dried over MgSO4, filtered and concentrated under reduced pressure to give a white amorphous solid, which was purified if needed.
(v) General Procedure Q for Suzuki-Miyaura Coupling:
To a suspension of Na2CO3 (3 equiv), the boronic acid (2.5 equiv), and the bromo-compound (1 equiv) in a mixture of toluene and H2O was added Pd(dppf)2Cl2.CH2Cl2 (0.2 equiv) and the suspension was heated under microwaves irradiation at 90° C. for 15 min or at 100° C. for 45 min. The mixture was filtered through a pad of Celite and washed with EtOAc. The residue on Celite was solubilized in MeOH and the filtrate was concentrated under reduced pressure. The residue was dissolved in CH2Cl2 and 1N HCl solution resulting to a precipitate which was filtered to give the desired compound, which was purified if needed.
(w) General Procedure R for Thio-Michael Addition:
To a solution of 134 (0.186 mmol, 1 equiv) in methanol was added a solution of arylthiol (01.2 equiv) and DIPEA (1.5 equiv) in methanol dropwise at 0° C. The solution was stirred at 0° C. for 30 min. Then the reaction mixture was diluted with EtOAc, washed twice with aq. K2CO3 solution, twice with saturated aq. NH4C1 solution, then with brine, dried over Na2SO4, evaporated under reduced pressure to afford the desired thiol, which was purified if necessary.
(x) General Procedure S for the Synthesis of N-Hydroxy Amidine:
Aminoacetonitrile hydrochloride (1 equiv) was placed in a flask where pyridine was carefully added to obtain a solution, and to this was added acyl chloride (1.05 eq) dropwise over 20 min. After stirring overnight at room temperature, water was carefully added; pyridinium hydrochloride dissolved while the product precipitated as a white solid. The precipitate was collected by filtration and washed with water. If no precipitation was observed, the mixture was extracted three times with dichloromethane. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give the cyanomethylbenzamide as a solid. To a solution of the obtained cyanomethylbenzamide (1.0 equiv) in methanol cooled to 0° C. were added hydroxylamine hydrochloride (1.0 equiv) and triethylamine (1.0 equiv), and the mixture was stirred at room temperature or heated to reflux overnight. The mixture was concentrated under reduced pressure. Water was added to the residue and the solid was collected by filtration, washed with water and dried to give the N-hydroxyamidine product, which was purified if necessary.
(y) General Procedure T for the Synthesis of Oxadiazole:
To a solution of the ester (1 equiv) in CH2Cl2 was added carbonyldiimidazole (2.2 equiv). The reaction mixture was stirred at room temperature for 10 min. N-hydroxyamidine (1 equiv) was added and the solution was stirred at room temperature for 16 h. The solvent was removed by evaporation in vacuo, and the residue was dissolved in EtOAc and washed twice with water. The organic layer was dried (MgSO4) and the solvent was removed by evaporation in vacuo. The residue was solubilized in DMF and the solution was heated to reflux for 4 h. The reaction mixture was cooled to room temperature, the solvent was removed by evaporation in vacuo, and the residue was dissolved in EtOAc and washed twice with water. The organic layer was dried (MgSO4) and the solvent was removed by evaporation in vacuo to afford theoxadiazole, which was purified if necessary.
2. Synthesis of Azide Precursors
methyl 3-azido-4-phenylbutanoate (106). Compound 106 was obtained according to general procedure H followed by general procedure A as a colorless oil (1.01 g, 71%). Purity: 100%, LC tR=5.83 min (method B), MS (ESI+): m/z=192 (M−N2+H)+. 1H NMR (CDCl3) δ (ppm): 7.37-7.22 (m, 5H), 4.12-4.03 (m, 1H), 3.71 (s, 3H), 2.90 (dd, J=7.4 and 13.7 Hz, 1H), 2.84 (dd, J=6.6 and 13.7 Hz, 1H), 2.55 (dd, J=5.3 and 16.1 Hz, 1H) 2.47 (dd, J=8.2 and J=16.1 Hz, 1H).
methyl (R)-3-azido-4-(naphthalen-2-yl)butanoate (107) Compound 107 was obtained according to general procedure H followed by general procedure A as an orange oil (622 mg, 74%), Purity: 100%, LC tR=6.54 min (method B), MS (ESI−): m/z=242 (M−N2−H)−. 1H NMR (CDCl3) δ (ppm): 7.86-7.80 (m, 3H), 7.70 (sl, 1H), 7.52-7.45 (m, 2H), 7.37 (dd, J=1.7 and 8.4 Hz, 1H), 4.24-4.15 (m, 1H), 3.71 (s, 3H), 3.08 (dd, J=13.7 and 7.3 Hz, 1H), 3.01 (dd, J=13.7 and 6.6 Hz, 1H), 2.59 (dd, J=16.1 and 5.4 Hz, 1H), 2.52 (dd, J=16.1 and 7.9 Hz, 1H).
3-azido-N-hydroxy-4-phenylbutanamide (108). Compound 108 was obtained according to general procedure E from azido ester 106 as an orange oil (75 mg, 25%), Purity: 100%, LC tR=4.04 min (method B), MS (ESI−): m/z=219 (M−H)—, 1H NMR (CD3OD) δ (ppm): 7.38-7.24 (m, 5H), 4.11-4.02 (m, 1H), 2.91 (dd, J=5.3 Hz and J=13.8 Hz, 1H), 2.79 (dd, J=8.4 Hz and J=13.8 Hz, 1H), 2.34 (dd, J=4.7 Hz and J=14.5 Hz, 1H), 2.22 (dd, J=9.1 Hz and J=14.5 Hz, 1H). 13C NMR (CD3OD) δ (ppm): 168.2, 137.3, 129.1, 128.2, 126.5, 60.8, 40.3, 37.2.
(3R)-3-azido-N-hydroxy-4-naphthylbutanamide (109). Compound 109 was obtained according to general procedure E from azido ester 107 as an orange oil (144 mg, 29%), Purity: 90%, LC tR=5.25 min (method B), MS (ESI+): m/z=271 [M+H]+. 1H NMR (CD3OD) δ (ppm): 7.84-7.80 (m, 3H), 7.74 (sl, 1H), 7.47-7.40 (m, 3H), 4.20 (dddd, J=4.7, 5.3, 8.2 Hz and 9.1 Hz, 1H), 3.08 (dd, J=5.3 Hz and J=13.7 Hz, 1H), 2.98 (dd, J=8.2 Hz and J=13.7 Hz, 1H), 2.38 (dd, J=4.7 Hz and J=14.6 Hz, 1H) 2.28 (dd, J=9.1 Hz and J=14.6 Hz, 1H). 13C NMR (CD3OD) δ (ppm): 168.3, 134.8, 133.6, 132.5, 127.9, 127.8, 127.3, 127.2, 125.8, 125.4, 60.7, 40.4, 37.3.
(R)-3-azido-4-(naphthalen-2-yl)butanoic acid (110). Compound 110 was obtained according to general procedure A as a yellow oil (2.1 g, quantitative yield), LC tR=2.84 min (method A), MS (ESI−): m/z=254 (M−N2+H)+. 1H NMR (CDCl3) δ (ppm): 7.85-7.80 (m, 3H), 7.69 (s, 1H), 7.52-7.45 (m, 2H), 7.37 (dd, J=1.5 and 8.4 Hz, 1H), 4.22-4.13 (m, 1H), 3.11 (dd, J=7.2 and 13.8 Hz, 1H), 7.37 (dd, J=6.8 and 13.5 Hz, 1H), 2.66-2.51 (m, 2H). 13C NMR (CDCl3) δ (ppm): 175.7, 134.0, 133.5, 132.5, 128.5, 128.2, 127.7, 127.6, 127.3, 126.3, 125.9, 59.8, 40.7, 38.5.
(S)-3-azido-4-(naphthalen-2-yl)butanoic acid (111). Compound 111 was obtained according to general procedure A as a yellow solid (902 mg, quantitative yield), LC tR=3.12 min (method A), MS (ESI−): m/z=254 (M−H)−. 1H NMR (CDCl3) δ (ppm): 7.85-7.81 (m, 3H), 7.69 (s, 1H), 7.52-7.45 (m, 2H), 7.37 (dd, J=1.8 and 8.4 Hz, 1H), 4.22-4.13 (m, 1H), 3.11 (dd, J=7.2 and 13.5 Hz, 1H), 3.02 (dd, J=6.6 and 13.5 Hz, 1H), 2.61-2.57 (m, 2H). 13C NMR (CDCl3) δ (ppm): 176.3, 134.0, 133.5, 132.5, 128.5, 128.2, 127.7, 127.6, 127.3, 126.3, 125.9, 59.8, 40.7, 38.6.
(2R)-2-Azido-3-naphthalen-2-yl-propionic acid (130). Compound 130 was obtained according to general procedure A as a brown solid (764 mg, 79%), Purity: 100%, LC tR=2.39 min (method A), MS (ESI−): m/z=240 [M−H]−. 1H NMR (CDCl3) δ (ppm): 7.83-7.81 (m, 3H), 7.72 (s, 1H), 7.51-7.47 (m, 2H), 7.39-7.37 (dd, J=1.5 and 8.4 Hz, 1H), 4.25 (dd, J=4.8 and 8.7 Hz, 1H), 3.40 (dd, J=4.5 and 13.8 Hz, 1H), 3.19 (dd, J=9 and 14.1 Hz, 1H). 13C NMR (CDCl3) δ (ppm): 133.5, 133.1, 132.6, 128.6, 128.2, 127.8, 127.1, 126.4, 126.0, 57.0, 37.7.
Methyl (R)-3-azido-4-(1H-indol-3-yl)butanoate (131). Compound 131 was obtained according to general procedure H followed by general procedure A as a yellow oil (74 mg, 77%), Purity: 94%, LC tR=2.83 min (method A), MS (ESI+): m/z=231 [M−N2+H]+. 1H NMR (CDCl3) δ (ppm): 8.09 (br s, 1H), 7.60 (d, J=8.0 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.17 (m, 3H), 4.16 (m, 1H), 3.67 (s, 3H), 3.04 (m, 2H), 2.54 (m, 2H). 13C NMR (DMSO-d6) δ (ppm): 172.0, 135.9, 126.5, 123.4, 120.8, 117.7, 118.3, 111.1, 100.2, 51.1, 48.8, 31.7, 21.1.
Methyl (R,E)-3-azido-6-phenylhex-5-enoate (139). Compound 139 was obtained according to general procedure H followed by general procedure A as a colorless oil (104 mg, 39%), Purity: 90%, LC tR=3.23 min (method A), MS (ESI+): m/z=218 [M−N2+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.39-7.17 (m, 5H), 6.52 (d, J=15.7 Hz, 1H), 6.23 (dt, J=7.3 and 15.7 Hz, 1H), 4.00-3.91 (m, 1H), 3.68 (s, 3H), 2.63 (dd, J=4.6 and 16.1 Hz, 1H), 2.52-2.44 (m, 3H).
Methyl (R)-3-azido-4-(naphthalen-1-yl)butanoate (147). Compound 147 was obtained according to general procedure H followed by general procedure A as a yellow powder (275 mg, quant. yield), Purity: 96%, LC tR=1.90 min (method A), MS (ESI+): m/z=242 [M−N2+H]+. 1H NMR (MeOD-d4) δ (ppm): 8.14 (d, J=8.1 Hz, 1H), 7.90 (d, J=8.1 Hz, 1H), 7.80 (dt, J=4.8 Hz, 1H), 7.59-7.47 (m, 2H), 7.42 (d, J=4.8 Hz, 2H), 4.19 (dddd, J=5.4, 8.5, 10.5 and 13.8 Hz, 1H), 3.67 (s, 3H), 3.40 (dd, J=8.5 Hz, J=13.9 Hz, 2H), 2.60 (m, 2H). 13C NMR (MeOD-d4) δ (ppm): 172.9, 135.5, 134.7, 133.3, 129.9, 129.1, 128.9, 127.3, 126.7, 126.4, 124.5, 61.3, 52.3, 40.0, 38.7.
Methyl (R)-3-azido-4-(4-chlorophenyl)butanoate (148). Compound 148 was obtained according to general procedure H followed by general procedure A as a yellow powder (59 mg, 93%), Purity: 80%, LC tR=3.13 min (method A), MS (ESI+): m/z=226 [M−N2+H]+.
Methyl (R)-3-azido-5-phenylpentanoate (149). Compound 149 was obtained according to general procedure H followed by general procedure A as a yellow powder (81 mg, 54%), Purity: 95%, LC tR=3.18 min (method A), MS (ESI+): m/z=206 [M−N2+H]+.
Methyl (R)-3-azido-5-phenylpentanoate (150). Compound 150 was obtained according to general procedure H followed by general procedure A as a yellow powder (55 mg, quant. yield), Purity: 90%, LC tR=3.18 min (method A), MS (ESI+): m/z=260 [M-N2+1-1]+.
Methyl (3R)-3-azido-2-hydroxy-4-(naphthalen-2-yl)butanoate (119). (2R)-2-(tert-butoxycarbonylamino)-3-(2-naphthyl)propanoic acid (0.960 g, 3.04 mmol) was dissolved with dry CH2Cl2 (7 mL, 0.42 M) and the solution was cooled to −15° C. N,O-Dimethylhydroxylamine hydrochloride (0.306 g, 3.13 mmol) was added, followed by N-methylmorpholine (0.34 mL, 3.06 mmol)). After 5 min, 1-(3-methylaminopropyl-3-ethylcarbodiimide hydrochloride (0.578 g, 3.06 mmol)) was added in five portions over 30 min. The reaction was warmed to room temperature overnight. Water (1 mL) was added and the solution was extracted with CH2Cl2 (3×2.5 mL). The combined organic phases were washed with brine (2.5 mL), dried on Na2SO4, and concentrated. The residue was purified by flash chromatography (cyclohexane/AcOEt (100/0 to 70/30) to give 115 as a white solid (1.08 g, 95%). Purity: 100%, LC tR=3.10 min, MS (ESI+): m/z=359 [M+H]+. 1H NMR (CDCl3) δ (ppm): 1.36 (s, 9H), 3.00-3.26 (m, 5H), 3.65 (s, 3H), 5.03 (br s, 1H), 5.20 (br s, 1H, NH), 7.31 (d, J=8.5 Hz, 1H), 7.42-7.46 (m, 2H), 7.62 (s, 1H), 7.76-7.81 (m, 3H). 13C NMR (75 MHz, CDCl3) δ (ppm): 28.2, 28.5, 32.3, 39.2, 51.7, 61.8, 79.8, 125.7, 126.2, 127.8, 127.8, 128.1, 128.3, 132.6, 133.7, 134.3, 172.6, 207.1. In inert atmosphere tert-butyl N-[(1R)-2-[methoxy(methyl)amino]-1-(2-naphthylmethyl)-2-oxo-ethyl]carbamate (115) (0.750 g, 2.092 mmol) in anhydrous Et2O (8.05 mL). The solution was cooled to −33° C. and LiAlH4 (2.5 mL, 2.5 mmol, 1M in THF) was added dropwise, and the reaction mixture was warmed to 0° C. After 45 min, the solution was cooled to −33° C. and 10% aq KHSO4 (4.03 mL) was added to quench the reaction. After warming to room temperature, 1N HCl (4.03 mL) was added, and the organic phases were separated, the aqueous phase was extracted with EtOAc (2×4 mL), and the combined organic phases were dried over MgSO4, filtered, and concentrated. The residue was purified by flash chromatography on silica gel (cyclohexane/AcOEt: 100/0 to 0/100 during 60 min) to yield quantitatively the aldehyde 116 (0.626 g) as a colorless solid. Purity: 100%, LC tR=2.95 min and 3.45 min, MS (ESI+): m/z=300 [M+H]+. 1H NMR (CDCl3) δ (ppm): 1.43 (s, 9H), 3.29 (d, J=6.5 Hz, 2H), 4.50 (q, J=6.5 Hz, J=13.0 Hz, 1H), 5.06 (br s, 1H, NH), 7.30 (dd, J=1.8 Hz, J=8.4 Hz, 1H), 7.47 (ddd, J=1.9 Hz, J=5.7 Hz, J=9.5 Hz, 2H), 7.62 (br s, 1H), 7.76-7.84 (m, 3H). 13C NMR (CDCl3) δ (ppm): 28.4, 35.8, 60.9, 126.0, 126.5, 127.4, 127.7, 127.8, 128.2, 128.7, 132.6, 133.4, 133.6, 155.4, 199.6. To a solution of tert-butyl N-[(1R)-1-formyl-2-(2-naphthyl)ethyl]carbamate 116 (361 mg, 1.21 mmol) in THF (3.65 mL) at 0° C. was added aqueous solution of NaHSO3 (150.6 mg, 1.447 mmol, in 4.385 ml of water) The mixture was stirred at 0° C. overnight. KCN (78.5 mg, 1.21 mmol) was added to the mixture and was stirred for 3 h at room temperature. The mixture was extracted with EtOAc (2×20 mL) and the collected organic layer were dried over Na2SO4, filtered and evaporated to afford the desired compound (468 mg). To a solution of the obtained compound (0.393 mg, 1.206 mmol) in dioxane (4.5 mL) was added a concentrated solution of HCl (6N, 8.9 mL). The solution was gently refluxed and stirred overnight. Then the solution was cold to room temperature and concentrated in vacuo. The crude was purified by preparative HPLC (MeCN/H2O, 0.1% HCO2H, gradient from 2% to 100% MeCN over 30 min) to afford the diastereoisomer 117 as a white solid (0.218 g, 74%). LC tR=1.75 and 1.68 min, MS (ESI+): m/z=246 [M+H]+. 118 was obtained according to general procedure H from 117. White powder. LC tR=1.85 and 1.95 min, MS (ESI+): m/z=260 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.88 (dd, J=7.9 Hz, J=13.8 Hz, 1H), 2.96 (dd, J=7.0 Hz, J=13.2 Hz, 1H), 3.05 (m, 2H), 3.54 (s, 3H), 3.51-3.57 (m, 1H), 3.60-3.67 (m, 1H), 3.71 (s, 3H), 4.06 (d, J=2.8 Hz, 1H), 4.28 (d, J=3.6 Hz, 1H), 7.37-7.50 (m, 6H), 7.72 (s, 1H), 7.73 (s, 1H), 7.79-7.85 (m, 6H). Azide 119 was obtained according to general procedure A from 118. Transparent oil. LC tR=3.15 and 3.25 min. 1H NMR (MeOD) δ (ppm): 3.11 (dd, J=2.6 Hz, 1H), 3.13 (d, J=4.0 Hz, 1H), 3.30 (d, J=2.6 Hz, 2.8H), 3.74 (s, 2.8H), 3.80 (s, 4.4H), 3.87 (td, J=2.0 Hz, J=7.8 Hz, 1.5H), 3.99 (ddd, J=3.0 Hz, J=6.7 Hz, J=8.0 Hz, 1H), 4.15 (d, J=1.9 Hz, 1.4H), 4.39 (d, J=2.2 Hz, 1H), 7.37 (dd, J=1.7 Hz, J=8.5 Hz, 1H), 7.42 (dd, J=1.8 Hz, J=8.5 Hz, 1.7H), 7.47-7.52 (m, 4H), 7.70 (br s, 1H), 7.77 (br s, 1.5H), 7.80-7.85 (m, 6H).
(S)-3-tert-Butoxycarbonylamino-4-hydroxy-butyric acid methyl ester (120). To a stirred solution of N-Boc aspartic acid O-methyl ester (4.15 g, 16.8 mmol, 1 equiv) in ethyl acetate (33 mL) was added solid N-hydroxysuccinimide (2.07 g, 18.0 mmol, 1.07 equiv) at 0° C. A solution of dicyclohexylcarbodiimide (3.52 g, 17.1 mmol, 1.02 equiv) in ethyl acetate (33 mL) was added slowly. The reaction was allowed to attain room temperature and stirred overnight. The precipitate of dicyclohexylurea was filtered off, and the filtrate was washed successively with saturated aqueous sodium hydrogen carbonate and brine, dried, and evaporated under reduced pressure to give crude succinimide ester (5.78 g, quantitative yield). Purity: 83%, LC tR=2.39 min, MS (ESI+): m/z=345 [M+1-1]+. Sodium borohydride (1.03 g, 27.4 mmol, 1.6 equiv) was dissolved in water (7 mL) and THF (50 mL) at 0° C. A solution of the obtained succinimide ester (5.78 g, 16.8 mmol, 1 equiv) in THF (8 mL) was added dropwise. The reaction was stirred for 15 min. Saturated aqueous ammonium chloride was added to quench the reaction. Extraction with ethyl acetate followed by washing of the combined organic extracts with brine, drying, and evaporation to dryness, afforded the crude product, which was purified by flash chromatography on silica gel (petroleum ether/EtOAc 1:0 to 1:1) to give the desired compound 120 as a colorless oil (3.14 g, 40%). Purity: 50%, LC tR=1.97 min, MS (ESI+): m/z=256 [M+Na]+. 1H NMR (CDCl3) δ (ppm): 5.27 (br s, 1H), 3.99-3.93 (m, 1H), 3.69-3.68 (m, 5H), 2.77 (br s, 1H), 2.63 (d, J=6.0 Hz, 2H), 1.43 (s, 9H).
Methyl (3S)-3-(tert-butoxycarbonylamino)-4-oxo-butanoate (121). A solution of Des s-Martin periodinane (1.15 equiv) in anhydrous CH2Cl2 was added at room temperature to a solution of 120 (1 equiv) in anhydrous CH2Cl2. The mixture was stirred during 1 h15. The mixture was diluted with CH2Cl2 and the excess reagent was quenched with a saturated aqueous solution of NaHCO3 containing Na2S2O3. The phases were separated and the organic layer washed with saturated aqueous solution of NaHCO3, H2O, dried over MgSO4, filtered and evaporated to give the crude product, which was purified by flash chromatography on silica gel (cyclohexante/EtOAc 100/0 to 70/30). Purity: 100%, LC tR=2.03 min, MS (ESI+): m/z=232 [M+H]+. 1H NMR (CDCl3) δ (ppm): 1.45 (s, 9H), 2.82 (dd, J=4.9 Hz, J=17.4 Hz, 1H), 2.99 (dd, J=4.7 Hz, J=17.4 Hz, 1H), 3.69 (s, 3H), 4.35 (ddd, J=4.7 Hz, J=4.9 Hz, J=6.7 Hz, 1H), 5.62 (d, J=6.7 Hz), 9.64 (s, 1H). 13C NMR (CDCl3; 75 MHz), δ (ppm): 28.4, 34.4, 52.3, 56.0, 80.7, 155.6, 171.7, 199.3.
Methyl (S)-3-azido-4-(3,4-dihydroisoquinolin-2(1H)-yl)butanoate (122). Compound 121 (1.5 equiv), 1,2,3,4-tetrahydroisoquinoline (1 equiv) and 3A molecular sieves were added in DCE. After stirring 15 min, sodium triacetoxyborohydride (2.2 equiv) was added in small portions. The mixture was stirred at room temperature under argon overnight. The reaction was diluted with CH2Cl2 and washed with water. The organic layer was extracted with CH2Cl2. Then the combined organic layer was washed with brine, dried over Na2SO4, filtered and evaporated to give the crude product, which was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 10/90). Azide 122 was obtained according to general procedure N from the obtained product followed by general procedure A. LC tR=1.77 min, MS (ESI+): m/z=275 [M+H]+. This compound was directly engaged in the next reaction.
(R)-3-tert-butoxycarbonylamino-4-iodo-butyric acid methyl ester (123) Triphenylphosphine (6.02 g, 23.0 mmol, 1.2 equiv), imidazole (1.56 g, 23.0 mmol, 1.2 equiv), and iodine (5.83 g, 23.0 mmol, 1.2 equiv) were added to dry dichloromethane (57 mL) with stirring. Alcohol 29 (4.46 g, 19.1 mmol, 1 equiv) was dissolved in dry dichloromethane (19 mL) under argon and transferred to the reaction mixture via syringe. The reaction was stirred at room temperature for 1.5 h. The mixture was filtered before washing with aqueous sodium thiosulfate solution (1M, 25 mL) and brine (25 mL) and drying (MgSO4). The dichloromethane was evaporated under reduced pressure, and then the residue was slurried in diethyl ether and filtered through a bed of silica, with washing with additional ether. The filtrate was concentrated under vacuum giving yellow oil. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc 1:0 to 1:1) giving a yellow solid (1.64 g, 25%). Purity: 98%, LC tR=3.05 min, MS (ESI+): m/z=344 [M+H]+. 1H NMR (CDCl3) δ (ppm): 1.45 (s, 9H), 2.64 (dd, 1H, J=6.3 and 16.5 Hz), 2.76 (dd, 1H, J=5.4 and 16.5 Hz), 3.43 (m, 2H), 3.71 (s, 3H), 3.92 (m, 1H), 5.08 (br d, 1H, J=6.6).
(R)-3-azido-4-(3-bromo-phenyl)-butyric acid (124). Compound 124 was obtained according to general procedure J from 123 and 1-bromo-3-iodo-benzene followed by general procedure N and then general procedure A as an orange oil (510 mg, 32% over 3 steps). LC tR=3.40 min. 1H NMR (CDCl3) δ (ppm): 7.43-7.40 (m, 2H), 7.24-7.16 (m, 2H), 4.11-4.02 (m, 1H), 3.73 (s, 3H), 2.83 (d, J=6.9 Hz, 2H), 2.53-2.51 (m, 2H).
(R)-3-azido-4-(4-bromo-phenyl)-butyric acid (125). Compound 125 was obtained according to general procedure J from 123 and 1-bromo-4-iodo-benzene followed by general procedure N and then general procedure A as a yellow oil (3.44 g, 38% over 3 steps). LC tR=3.62 min. 1H NMR (CDCl3) δ (ppm): 7.48 (d, J=8.1 Hz, 2H), 7.11 (d, J=8.1 Hz, 2H), 4.05 (t, J=6.8 Hz, 1H), 3.91 (s, 3H), 2.82 (d, J=6.8 Hz, 2H), 2.52 (d, J=6.8 Hz, 2H).
Methyl (R)-3-azido-4-(3-(trifluoromethyl)phenyl)butanoate (126). Compound 126 was obtained according to general procedure K from 123 and 1-iodo-3-(trifluoromethyl)benzene followed by general procedure N and then general procedure A as a colorless oil (41 mg, 15% over 3 steps). Purity: 99%, LC tR=3.20 min.
Methyl (R)-3-azido-4-(4-(2-(tert-butyl)-2H-tetrazol-5-yl)phenyl)butanoate (127). Compound 127 was obtained according to general procedure K from 123 and 2-tert-butyl-5-(4-iodophenyl)-2H-1,2,3,4-tetrazole followed by general procedure N and then general procedure A as a colorless oil (41 mg, 26% over 3 steps). Purity: 99%, LC tR=3.06 min. MS (ESI+): m/z=344 [M+H]+.
Methyl (3R)-3-azido-4-(3-quinolyl)butanoate (140). Compound 140 was obtained according according to general procedure M from 123 and 3-bromoquinoline followed by general procedure 0 and then general procedure A as a colorless solid (69 mg, 19% over 3 steps). Purity: 92%, LC tR=2.48 min, MS (ESI+): m/z=271 [M+H]+.
Methyl (3R)-3-azido-4-tetralin-6-yl-butanoate (141). Compound 141 was obtained according according to general procedure L from 123 and 6-bromotetraline followed by general procedure 0 and then general procedure A as a yellowish oil (44 mg, 12% over 3 steps). LC tr=3.42 min, MS (ESI+): m/z=246 [M−N2+H]+.
(3R)-3-azido-4-(6-tetralinyl)butanehydroxamic acid (142). Compound 142 was obtained according to general procedure L from cyclohexyl (3S)-3-(tert-butoxycarbonylamino)-4-iodo-butanoate and 6-bromotetraline followed by general procedure 0 and then general procedures G′ and A′ as a brownish wax (60 mg, 38% over 4 steps). LC tr=2.53 min, MS (ESI−): m/z=273 [M−H]−. 1H NMR, CDCl3+1% TMS, δ (ppm): 7.01-6.96 (m, 1H), 6.93-6.81 (m, 2H), 4.10-3.99 (m, 1H), 2.83-2.67 (m, 6H), 2.36 (dd, J=14.7, 3.6 Hz, 1H), 2.21 (dd, J=14.7, 9.3 Hz, 1H), 1.80-1.72 (m, 4H). 13C NMR, CDCl3+1% TMS, δ (ppm): 168.8, 137.5, 135.9, 133.5, 130.0, 129.4, 126.4, 60.3, 40.1, 37.5, 29.3, 29.0, 23.2, 23.1.
Methyl (3R)-3-azido-4-(7-quinolyl)-butanoate (151). Compound 151 was obtained according to general procedure L from 123 and 7-bromoquinoline followed by general procedure 0 and then general procedure A″ as a colourless solid (23 mg, 2.7% over 3 steps). LC tr=2.40 min, MS (ESI+): m/z=271 [M+H]+. 1H NMR (CDCl3+1% TMS) δ(ppm): 8.91 (s, 1H), 8.15 (br d, J=8.4 Hz, 1H), 7.96 (s, 1H), 7.80 (br d, J=8.4 Hz, 1H), 7.45 (dd, J=8.4 and 1.7 Hz, 1H), 7.41-7.37 (m, 1H), 4.26-4.15 (m, 1H), 3.71 (s, 3H), 3.07 (dd, J=11.2 and 7.1 Hz, 2H), 2.59-2.55 (m, 2H).
Methyl (3R)-3-azido-4-(2-quinolyl)-butanoate (152). Compound 152 was obtained according to general procedure L from 123 and 6-iodoquinoline followed by general procedure 0 and then general procedure A″ as a dark red oil (71 mg, 29% over 3 steps). LC tr=2.62 min, MS (ESI+): m/z=271 [M+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 8.10 (d, J=8.5 Hz, 1H), 8.06 (ddd, J=8.5 and 1.6 and 1.1 Hz, 1H), 7.79 (dd, J=8.1 and 1.2 Hz, 1H), 7.70 (ddd, J=8.4 and 6.9 and 1.5 Hz, 1H), 7.51 (ddd, J=8.1 and 6.9 and 1.1 Hz, 1H), 7.32 (d, J=8.4 Hz, 1H), 4.64-4.54 (m, 1H), 3.71 (s, 3H), 3.22 (d, J=6.9 Hz, 2H), 2.71 (dd, J=16.2 and 4.5 Hz, 1H), 2.60 (dd, J=16.2 and 8.8 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ (ppm): 171.1, 157.6, 147.9, 136.6, 129.7, 129.1, 127.6, 127.0, 126.3, 121.9, 58.6, 51.9, 43.3, 39.1.
Methyl (3R)-3-azido-4-(6-quinolyl)-butanoate (153). Compound 153 was obtained according to general procedure L from 123 and 6-iodoquinoline followed by general procedure O′ and then general procedure B as a light beige wax (29 mg, 19% over 3 steps). LC tr=2.25 min, MS (ESI+): m/z=289 [M+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 9.26 (br s, 1H), 7.04-7.00 (m, 2H), 6.81 (d, J=8.5 Hz, 1H), 4.08-3.98 (m, 1H), 3.72 (s, 3H), 2.96 (dd, J=8.0 and 7.0 Hz, 2H), 2.80 (d, J=7.0 Hz, 2H), 2.65 (d, J=8.0 Hz, 1H), 2.63 (d, J=9.0 Hz, 1H), 2.54 (dd, J=16.1 and 5.4 Hz, 1H), 2.47 (dd, J=16.2 and 8.0 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ (ppm): 172.2, 171.1, 136.4, 131.5, 128.9, 128.4, 123.9, 115.8, 60.2, 52.0, 40.0, 38.7, 30.6, 25.3.
Methyl (3R)-3-azido-4-(6-(1,1,4,4-tetramethyl-2,3-dihydronaphthyl)-butanoate (154). Compound 154 was obtained according to general procedure L from 123 and 6-iodo-1,1,4,4-tetramethyl-1,2,3,4-tetrahydronaphthalene followed by general procedure S and then general procedure A′ as a yellowish thick oil (159 mg, 56% over 3 steps). LC tr=3.65 min, MS (ESI+): m/z=302 [M−N2+H]+. 1H NMR, (CDCl3+1% TMS) δ (ppm): 7.24 (d, J=8.1 Hz, 1H), 7.13 (d, J=1.9 Hz, 1H), 6.96 (dd, J=8.1 and 1.9 Hz, 1H), 4.08-3.99 (m, 1H), 3.69 (s, 3H), 2.85 (dd, J=13.8 and 7.3 Hz, 1H), 2.76 (dd, J=13.8 and 6.6 Hz, 1H), 2.53 (dd, J=16.1 and 5.0 Hz, 1H), 2.45 (dd, J=16.1 and 8.5 Hz, 1H), 1.67 (s, 4H), 1.27 (s, 6H), 1.26 (s, 6H). 13C NMR (CDCl3+1% TMS) δ (ppm): 171.2, 145.1, 143.5, 133.5, 127.4, 126.8, 126.5, 60.1, 51.9, 40.3, 38.7, 35.1, 35.0, 34.2, 34.0, 31.8 (4C).
Methyl (3R)-3-azido-4-(5-benzothiophene)-butanoate (155). Compound 155 was obtained according to general procedure L from 123 and 5-iodo-benzothiophene followed by general procedure O′ and then general procedure A′ as a yellow thick oil (120 mg, 49% over 3 steps). LC tr=3.02 min, MS (ESI+): m/z=248 [M−N2+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 7.82 (d, J=8.3 Hz, 1H), 7.66 (d, J=1.4 Hz, 1H), 7.44 (d, J=5.4 Hz, 1H), 7.29 (d, J=5.4 Hz, 1H), 7.20 (dd, J=8.3 and 1.6 Hz, 1H), 4.17-4.07 (m, 1H), 3.69 (s, 3H), 3.01 (dd, J=13.9 and 7.5 Hz, 1H), 2.94 (dd, J=13.9 and 6.7 Hz, 1H), 2.55 (dd, J=16.2 and 5.3 Hz, 1H), 2.48 (dd, J=16.2 and 8.1 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ (ppm): 171.1, 140.0, 138.5, 132.8, 127.0, 125.7, 124.2, 123.6, 122.7, 60.3, 51.9, 40.5, 38.6.
Methyl (3R)-3-azido-4-(6-benzothiophene)-butanoate (156). Compound 156 was obtained according to general procedure L from 123 and 6-iodo-benzothiophene followed by general procedure O′ and then general procedure A′ as a colourless oil (69 mg, 26% over 3 steps). LC tr=3.00 min, MS (ESI+): m/z=248 [M−N2+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 7.75 (d, J=8.2 Hz, 1H), 7.71-7.70 (m, 1H), 7.39 (d, J=5.4 Hz, 1H), 7.29 (dd, J=5.4 and 0.6 Hz, 1H), 7.21 (dd, J=8.2 and 1.6 Hz, 1H), 4.16-4.06 (m, 1H), 3.68 (s, 3H), 2.99 (dd, J=13.9 and 7.5 Hz, 1H), 2.92 (dd, J=13.8 and 6.7 Hz, 1H), 2.54 (dd, J=16.2 and 5.4 Hz, 1H), 2.47 (dd, J=16.2 and 8.0 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ(ppm): 171.0, 140.2, 138.6, 132.9, 126.3, 125.7, 123.7, 123.6, 123.0, 60.3, 51.9, 40.6, 38.6.
Methyl (3R)-3-azido-4-(5-indole)-butanoate (157). Compound 157 was obtained according to general procedure L from 123 and 5-iodoindole followed by general procedure O′ and then general procedure A′ as a brown solid (86 mg, 29% over 3 steps). LC tr=2.67 min, MS (ESI+): m/z=231 [M−N2+H]+. 1H NMR (CDCl3+1% TMS) δ(ppm): 8.23 (br s, 1H), 7.45 (br s, 1H), 7.27 (d, J=8.3 Hz, 1H), 7.13 (t, J=2.8 Hz, 1H), 7.00 (dd, J=8.3 and 1.5 Hz, 1H), 6.49-6.47 (m, 1H), 4.14-4.04 (m, 1H), 3.66 (s, 3H), 3.01 (dd, J=13.7 and 7.1 Hz, 1H), 2.87 (dd, J=13.7 and 7.1 Hz, 1H), 2.54 (dd, J=16.1 and 4.6 Hz, 1H), 2.44 (dd, J=16.2 and 8.9 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ (ppm): 171.5, 134.9, 128.2, 127.8, 124.8, 123.3, 121.2, 111.2, 102.2, 60.7, 51.9, 40.7, 38.5.
Methyl (3R)-3-azido-4-(6-indole)-butanoate (158). Compound 158 was obtained according to general procedure L from 123 and 6-iodoindole followed by general procedure O′ and then general procedure A′ as a brownish-red solid (67 mg, 24% over 3 steps). LC tr=2.72 min, MS (ESI+): m/z=259 [M+H]+. 1H NMR (CDCl3+1% TMS) δ(ppm): 8.19 (br s, 1H), 7.57 (d, J=8.1 Hz, 1H), 7.17 (br s, 1H), 7.12 (dd, J=3.1 and 2.4 Hz, 1H), 6.95 (dd, J=8.1 and 1.4 Hz, 1H), 6.51-6.49 (m, 1H), 4.14-4.05 (m, 1H), 3.66 (s, 3H), 3.01 (dd, J=13.7 and 7.1 Hz, 1H), 2.88 (dd, J=13.7 and 6.9 Hz, 1H), 2.53 (dd, J=16.1 and 4.7 Hz, 1H), 2.44 (dd, J=16.2 and 8.7 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ(ppm): 171.4, 136.1, 130.4, 128.9, 124.5, 121.3, 120.8, 111.7, 102.4, 60.6, 51.9, 40.8, 38.6.
Methyl (3R)-3-azido-4-(5-(N-methyl)indole)-butanoate (159). Compound 159 was obtained according to general procedure L from 123 and 5-iodoindole followed by general procedure O′ and then general procedure A′ as a yellow thick oil (25 mg, 2.6% over 3 steps). LC tr=3.05 min, MS (ESI+): m/z=273 [M+H]+.
Methyl (3R)-3-azido-4-(6-(1,3-benzothiazole))-butanoate (160). Compound 160 was obtained according to general procedure L from 123 and 6-bromo-1,3-benzothiazole followed by general procedure 0 and then general procedure A′ as a colourless wax (15 mg, 8.3% over 3 steps). LC tr=2.53 min, MS (ESI+): m/z=277 [M+H]+. 1H NMR (CDCl3+1% TMS), δ (ppm): 8.98 (s, 1H), 8.10 (d, J=8.4 Hz, 1H), 7.84 (s, 1H), 7.39 (d, J=8.4 Hz, 1H), 4.19-4.09 (m, 1H), 3.71 (s, 3H), 3.01 (d, J=7.0 Hz, 2H), 2.55 (d, J=7.6 Hz, 2H).
Methyl (3R)-3-azido-4-(3-benzothiophene)-butanoate (161). Compound 161 was obtained according to general procedure L from 123 and 3-iodobenzothiophene followed by general procedure 0 and then general procedure A′ as a light-brown thick wax (91 mg, 15% over 3 steps). LC tr=3.03 min, MS (ESI+): m/z=248 [M−N2+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 7.88-7.84 (m, 1H), 7.79-7.75 (m, 1H), 7.43-7.32 (m, 2H), 7.25 (s, 1H), 4.20 (quintet, J=6.9 Hz, 1H), 3.69 (s, 3H), 3.10 (dd, J=6.9 and 0.7 Hz, 2H), 2.56 (d, J=6.9 Hz, 2H). 13C NMR (CDCl3+1% TMS) δ (ppm): 171.0, 140.4, 138.5, 131.3, 124.5, 124.4, 124.2, 123.0, 121.4, 58.5, 52.0, 38.9, 33.3.
Methyl (3R)-3-azido-4-(2-benzothiophene)-butanoate (162). Compound 162 was obtained according to general procedure L from 123 and 2-iodobenzothiophene followed by general procedure O′ and then general procedure A′ as a yellowish thick oil (17 mg, 7.0% over 3 steps). LC tr=3.05 min, MS (ESI+): m/z=248 [M−N2+H]+. 1H NMR (CDCl3+1% TMS) δ (ppm): 7.80-7.76 (m, 1H), 7.72-7.68 (m, 1H), 736-7.26 (m, 2H), 7.12 (d, J=0.6 Hz, 1H), 4.23-4.14 (m, 1H), 3.72 (s, 3H), 3.22-3.08 (m, 2H), 2.63 (dd, J=16.3 and 5.0 Hz, 1H), 2.54 (dd, J=16.3 and 8.3 Hz, 1H). 13C NMR (CDCl3+1% TMS) δ (ppm): 170.9, 139.8, 139.8, 139.6, 124.4, 124.1, 123.4, 123.2, 122.2, 59.5, 52.1, 38.6, 35.6.
(3R)-3-azido-4-(6-(1,4-benzodioxyl))butanehydroxamic acid (163). Compound 163 was obtained according to general procedure L from 123 and 6-iodobenzodioxane followed by general procedure 0 and then general procedure G′ and then general procedure A′ as a yellowish wax (88 mg, 72% over 4 steps). LC tr=1.97 min, MS (ESI−): m/z=277 [M−H]−.
Methyl 3-azidoacrylate (133). A solution of sodium azide (50 mmol, 5 equiv), borax (15 mmol, 1.5 equiv) and KH2PO4 (15 mmol, 1.5 equiv) in water (70 mL) was prepared in a crystallizer. Methyl prop-2-ynoate (10 mmol, 1 equiv) was added and the crystallizer was put into the ultrasound and sonicated at room temperature for 1 h. Then the mixture was extracted with Et2O, dried with Na2SO4 and concentrated at 25° C. under reduced pressure to give the desired compound as white crystals.
3. Triazole Formation
N-[1-((R)-2-Hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-benzamide (1). Compound 1 was obtained according to general procedure B from azide 109 and N-(prop-2-yn-1-yl)benzamide as a beige solid (24 mg, 51%), Purity: 95%, mp 192.8-194.0° C., LC tR=3.10 min, MS (ESI+): m/z=430 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (s, 1H), 8.95 (t, J=5.9 Hz, 0.8H), 8.79 (s, 0.5H), 7.88-7.67 (m, 5H), 7.56-7.35 (m, 7H), 7.15 (dd, J=1.3 and 8.6 Hz, 1H), 5.28-5.26 (m, 1H), 4.42 (d, J=5.6 Hz, 2H), 3.32 (m, 2H), 2.77 (dd, J=8.8 and 15.0 Hz, 1H), 2.65 (dd, J=5.4 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.5, 166.0, 145.1, 135.0, 134.6, 133.3, 132.3, 131.7 (2C), 128.7 (2C), 128.2, 127.9 (2C), 127.7 (3C), 126.5, 126.0, 122.9, 59.2, 41.4, 37.6, 35.2. HRMS m/z calculated for C24H24N5O3 [M+H]+ 430.1879, found 430.1884.
N-[1-(R)-2-Hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-4-methoxy-benzamide (2). Compound 2 was obtained according to general procedure B from azide 109 and 4-methoxy-N-(prop-2-yn-1-yl)benzamide as a beige solid (43 mg, 68%), Purity: 96%, mp 178.3-179.6° C., LC tR=2.33 min, MS (ESI+): m/z=460 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.50 (s, 1H), 8.80 (m, 2H), 7.86-7.69 (m, 6H), 7.47-7.39 (m, 3H), 7.15 (d, J=8.6 Hz, 1H), 6.98 (d, J=8.6 Hz, 2H), 5.26 (m, 1H), 4.40 (d, J=5.3 Hz, 2H), 3.81 (s, 3H), 3.31 (m, 2H), 2.77 (dd, J=9.5 and 15.1 Hz, 1H), 2.65 (dd, J=5.2 and 15.1 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0 (2C), 162.1, 145.2, 135.0, 133.3, 132.3, 129.6 (2C), 128.2 (2C), 127.9 (2C), 127.8, 126.8, 126.5, 126.0, 123.0, 113.9 (2C), 59.2, 55.8, 41.4, 37.6, 35.1. HRMS m/z calculated for C25H26N5O4 [M+H]+460.1985, found 460.1977.
4-Fluoro-N-[1-(R-2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]-triazol-4-ylmethyl]-benzamide (3). Compound 3 was obtained according to general procedure B from azide 109 and 4-fluoro-N-(prop-2-yn-1-yl)benzamide as a beige solid (36 mg, 57%), Purity: 96%, mp 183.6-184.1° C., LC tR=2.42 min, MS (ESI+): m/z=448 [1\4+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.50 (s, 1H), 8.99 (t, 0.9H, J=5.4 Hz, 1H), 8.80 (s, 1H), 7.93-7.87 (m, 3H), 7.78-7.68 (m, 3H), 7.45-7.36 (m, 3H), 7.29 (t, J=8.8 Hz, 2H), 7.14 (d, J=8.9 Hz, 1H), 5.28-5.24 (m, 1H), 4.41 (d, J=5.4 Hz, 2H), 3.31 (m, 2H), 2.77 (dd, J=8.7 and 14.9 Hz, 1H), 2.66 (dd, J=5.4 and 14.9 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 165.5, 165.0, 163.9 (d, JC-F=247 Hz), 144.5, 135.0, 132.9, 132.3, 131.8, 130.7, 130.6, 130.0, 129.9, 127.7, 127.4 (2C), 127.3, 126.0, 125.6, 118.0, 115.2 (d, JC-F=22 Hz), 58.8, 40.9, 37.1, 34.8. 19F NMR (DMSO-d6) δ (ppm): −109.99. HRMS m/z calculated for C24H23FN5O3 [M+H]+ 448.1785, found 448.1801.
N-[1-(R-2-Hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-4-trifluoromethyl-benzamide (4). Compound 4 was obtained following general procedure B from azide 109 and N-(prop-2-yn-1-yl)-4-(trifluoromethyl)benzamide as a beige solid (36 mg, 40%), Purity: 95%, mp 198.9-200.4° C., LC tR=2.62 min, MS (ESI+): m/z=498 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (s, 1H), 9.20 (s, 1H), 8.80 (s, 1H), 8.02 (m, 2H), 7.88-7.83 (m, 3H), 7.76-7.65 (m, 3H), 7.46-7.38 (m, 3H), 7.12 (d, J=8.8 Hz, 1H), 5.27 (m, 1H), 4.44 (d, J=3.9 Hz, 2H), 3.32 (m, 2H), 2.82-2.64 (m, 2H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.3, 144.7, 138.3, 134.9, 133.3, 132.2, 131.6 (d, JC-F=30 Hz), 128.7, 128.2, 127.8, 127.7, 126.0, 125.8, 124.5 (q, JC-F=271 Hz), 123.1, 59.3, 41.4, 37.6, 35.3. HRMS m/z calculated for C25H22O3N5F3 [M+H]+ 498.1753, found 498.1749.
(R)—N-Hydroxy-4-naphthalen-2-yl-3-{4-[(toluene-4-sulfonylamino)-methyl]-[1,2,3]triazol-1-yl}-butyramide (5). Compound 5 was obtained according to general procedure B from azide 109 and 4-methyl-N-(prop-2-yn-1-yl)benzenesulfonamide as a beige solid (20 mg, 35%), Purity: 95%, mp 115.8-117.2° C., LC tR=2.55 min, MS (ESI+): m/z=480 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (s, 1H), 8.81 (s, 1H), 8.00 (t, J=6.0 Hz, 1H), 7.85-7.75 (m, 4H), 7.65 (d, J=8.2 Hz, 2H), 7.47-7.42 (m, 3H), 7.33 (d, J=8.2 Hz, 2H), 7.14 (d, J=7.4 Hz, 1H), 5.25-5.21 (m, 1H), 3.92 (d, J=5.8 Hz, 2H), 3.27 (m, 2H), 2.71 (dd, J=8.9 and 15.2 Hz, 1H), 2.69 (dd, J=5.7 and 15.2 Hz, 1H), 2.34 (s, 3H). 13C NMR (DMSO-d6) δ (ppm): 165.9, 143.4, 143.1, 137.9, 135.0, 133.4, 132.3, 130.0, 128.3, 127.9, 127.7, 127.1, 126.6, 126.1123.2, 59.4, 41.3, 38.5, 37.6, 21.4. HRMS m/z calculated for C24H25N5O4S [M+H]+ 480.1706, found 480.1696.
(R)-3-{4-[(4-Fluoro-benzenesulfonylamino)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (6). Compound 6 was obtained according to general procedure B from azide 109 and 4-fluoro-N-(prop-2-yn-1-yl)benzenesulfonamide as a beige solid (40 mg, 60%), Purity: 95%, mp 157.1-158.7° C., LC tR=2.54 min, MS (ESI+): m/z=484 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 7.83-7.75 (m, 6H), 7.54-7.33 (m, 5H), 7.15 (d, J=8.7 Hz, 1H), 5.27-5.23 (m, 1H), 3.95 (s, 2H), 3.28-3.26 (m, 2H), 2.75 (dd, J=9.1 and 14.9 Hz, 1H), 2.63 (dd, J=5.6 and 14.9 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 164.5 (d, JC-F=247 Hz), 137.1, 135.0, 133.4, 132.3, 130.1 (2C), 130.0, 128.3, 127.9 (3C), 127.7, 126.5, 126.1, 123.3, 116.7 (d, JC-F=22 Hz, 2C), 59.3, 41.3, 38.4, 37.5. HRMS m/z calculated for C23H23FN5O4S [M+H]+ 484.1455, found 484.1457.
(R)-3-[4-(Benzenesulfonylamino-methyl)-[1,2,3]triazol-1-yl]-N-hydroxy-4-naphthalen-2-yl-butyramide (7). Compound 7 was obtained according to general procedure B from azide 109 and N-(prop-2-yn-1-yl)benzenesulfonamide as a beige solid (25 mg, 47%), Purity: 96%, mp 137.4-138.8° C., LC tR=2.47 min, MS (ESI+): m/z=466 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (s, 1H), 8.80 (s, 1H), 8.09 (m, 1H), 7.82-7.76 (m, 6H), 7.57-7.46 (m, 6H), 7.14 (d, J=8.1 Hz, 1H), 5.24 (m, 1H), 3.93 (m, 2H), 3.28 (3, 2H), 2.70-2.66 (m, 2H). 13C NMR (DMSO-d6) δ (ppm): 165.9, 140.7, 135.0, 133.4, 132.9, 132.3, 129.6, 128.3, 127.9, 127.7, 127.0, 126.6, 126.1, 59.3, 41.3, 38.6, 37.6. HRMS m/z calculated for C23H23N5O4S [M+H]+ 466.1549, found 466.1550.
(R)—N-Hydroxy-4-naphthalen-2-yl-3-{4-[(2-p-tolyl-acetylamino)-methyl]-[1,2,3]triazol-1-yl}-butyramide (8). Compound 8 was obtained according to general procedure B from azide 109 and N-(prop-2-yn-1-yl)-2-(p-tolyl)acetamide as a yellow oil (21 mg, 55%), Purity: 90%, LC tR=2.52 min, MS (ESI+): m/z=458 [M+1-1]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (s, 1H), 8.81 (s, 1H), 8.41 (t, J=5.0 Hz, 1H), 7.85-7.74 (m, 4H), 7.49-7.44 (m, 3H), 7.15-7.04 (m, 5H), 5.26 (m, 1H), 4.20 (d, J=5.2 Hz, 2H), 3.31-3.25 (m, 4H), 2.76 (dd, J=9.3 and 15.3 Hz, 1H), 2.65 (dd, J=5.4 and 15.3 Hz, 1H), 2.24 (s, 3H). 13C NMR (DMSO-d6) δ (ppm): 170.7, 166.1, 144.8, 135.9, 135.1, 133.7, 133.4, 132.4, 129.4, 129.3 (2C), 128.3, 127.9 (3C), 127.7 (2C), 126.6, 126.2, 122.9, 59.3, 42.2, 41.6, 37.8, 34.7, 21.1. HRMS m/z calculated for C26H28N5O3 [M+H]+ 458.2192, found 458.2212.
(R)-3-{4-[(4-Fluoro-benzenesulfonylamino)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (9). Compound 9 was obtained according to general procedure B from azide 109 and pent-4-yn-1-ylbenzene as a beige solid (40 mg, 60%), Purity: 95%, mp 157.1-158.7° C., LC tR=2.54 min, MS (ESI+): m/z=484 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 7.83-7.75 (m, 6H), 7.54-7.33 (m, 5H), 7.15 (d, J=8.7 Hz, 1H), 5.27-5.23 (m, 1H), 3.95 (s, 2H), 3.28-3.26 (m, 2H), 2.75 (dd, J=9.1 and 14.9 Hz, 1H), 2.63 (dd, J=5.6 and 14.9 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 164.5 (d, JC-F=247 Hz), 137.1, 135.0, 133.4, 132.3, 130.1, 130.0, 128.3, 127.9, 127.7, 126.5, 126.1, 116.7 (d, JC-F=22 Hz), 59.3, 41.3, 38.4, 37.5. HRMS m/z calculated for C23H23FN5O4S [M+H]+484.1455, found 484.1457.
(R)—N-Hydroxy-4-naphthalen-2-yl-3-{4-[(3-phenyl-propionylamino)-methyl]-[1,2,3]triazol-1-yl}-butyramide (10). Compound 10 was obtained according to general procedure B from azide 109 and 3-phenyl-N-(prop-2-yn-1-yl)propanamide as a beige solid (43 mg, 82%), Purity: 95%, mp 150.0-151.4° C., LC tR=2.50 min, MS (ESI+): m/z=457 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (s, 1H), 8.81 (s, 1H), 8.27 (m, 1H), 7.83-7.67 (m, 4H), 7.50-7.42 (m, 3H), 7.28-7.16 (m, 6H), 5.25 (m, 1H), 4.20 (m, 2H), 3.32 (m, 2H), 2.80-2.73 (m, 2H), 2.78 (t, J=7.5 Hz, 2H), 2.35 (t, J=7.8 Hz, 2H). 13C NMR (DMSO-d6) δ (ppm): 171.6, 166.0, 141.8, 135.3, 135.1, 133.4, 132.3, 128.7 (2C), 128.6 (2C), 128.3, 127.9 (3C), 127.7, 126.5, 126.3 (2C), 126.1, 59.2, 41.4, 37.7, 37.3, 34.5, 31.5. HRMS m/z calculated for C26H28N5O3 [M+H]+ 458.2192, found 458.2191.
(R)—N-[1-(2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-N-methyl-benzamide (14). Compound 14 was obtained according to general procedure C from azide 110 and N-methyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure D as a white solid (0.049 g, 37%). Purity: 100%, LC tR=2.32 min (method A), MS (ESI+): m/z=444 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.54 (s, 0.9H), 8.81 (m, 0.8H), 7.87-7.70 (m, 4H), 7.46-7.15 (m, 10H), 5.33-5.24 (m, 1H), 4.48 (q, J=15.3 Hz, 1H), 4.30 (s, 1H), 3.41-3.16 (m, 5H), 2.78 (d, J=5.7 Hz, 1H), 2.68 (s, 1H). 13C NMR (CD3CN) δ (ppm): 171.1, 166.7, 142.8, 134.6, 133.3, 132.3, 129.6, 128.3, 127.9, 127.7, 127.5, 127.4, 127.2, 126.8, 126.1, 125.7, 59.7, 41.1, 37.6, 36.5, 31.9. HRMS m/z calculated for C25H26N5O3 [M+H]+ 444.2036, found 444.2036.
(R)-4,4-difluoro-cyclohexanecarboxylic acid [1-(2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-amide (15). Compound 15 was obtained according to general procedure C from azide 110 and 4,4-difluoro-N-(prop-2-yn-1-yl)cyclohexane-1-carboxamide followed by general procedure D as a white solid (0.016 g, 10%). Purity: 96%, LC tR=2.35 min (method A), MS (ESI+): m/z=472 [M+H]+. 1H NMR (CD3CN) δ (ppm): 8.98 (m, 1H), 7.85-7.75 (m, 3H), 7.49-7.46 (m, 3H), 7.41 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 6.95 (s, 1H), 6.75 (s, 1H), 5.29-5.19 (m, 1H), 4.25-4.28 (m, 2H), 3.41-3.38 (m, 2H), 2.82-2.78 (m, 2H), 1.57-1.83 (m, 6H). 13C NMR (MeOD-d4) δ (ppm): 175.8, 167.1, 144.2, 134.1, 133.5, 132.5, 129.1, 127.9, 127.4, 127.2, 126.6, 125.8, 125.4, 123.3, 122.5, 60.0, 42.0, 41.0, 37.6, 33.9, 32.4 (t, JC-F=24.1 Hz), 25.4 (dd, JC-F=6.0 and 9.2 Hz). HRMS m/z calculated for C24H28N5O3F2 [M+H]+ 472.2160, found 472.2180.
(R)—N-hydroxy-4-naphthalen-2-yl-3-[4-(phenylacetylamino-methyl)-[1,2,3]triazol-1-yl]-butyramide (16). Compound 16 was obtained according to general procedure C from azide 110 and 2-phenyl-N-(prop-2-yn-1-yl)acetamide followed by general procedure D as a white solid (36 mg, 30%). Purity: 95%, LC tR=2.34 min, MS (ESI+): m/z=444 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.08 (s, 1H), 7.85-7.74 (m, 3H), 7.48-7.42 (m, 4H), 7.34 (m, 6H), 6.88 (s, 1H), 5.28-5.17 (m, 1H), 4.25 (d, J=5.7 Hz, 2H), 3.42-3.35 (m, 4H), 2.81-2.76 (m, 2H). 13C NMR (CD3CN) δ (ppm): 170.9, 166.4, 144.5, 135.9, 134.6, 134.5, 133.4, 132.3, 129.2, 128.5 (2C), 128.0, 127.7, 127.5 (2C), 127.2, 126.7, 126.2, 125.8, 122.4, 59.5, 42.6, 40.9, 37.7, 34.7. HRMS m/z calculated for C25H26N5O3 [M+H]+ 444.2036, found 444.2024.
(R)-3-{4-[(2-ethyl-butyrylamino)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (17). Compound 17 was obtained according to general procedure C from azide 110 and 2-ethyl-N-(prop-2-yn-1-yl)butanamide followed by general procedure D as a white solid (0.009 g, 7%). Purity: 98%, LC tR=2.37 min (method A), MS (ESI+): m/z=424 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.01 (s, 0.8H), 7.84-7.74 (m, 3H), 7.52-7.45 (m, 4H), 7.20 (dd, J=1.8 and 8.4 Hz, 1H), 6.82 (s, 0.9H), 5.31-5.21 (m, 1H), 4.28 (dd, J=2.1 and 5.7 Hz, 2H), 3.40 (d, J=7.5 Hz, 2H), 1.83-1.91 (m, 1H), 2.81-2.76 (m, 2H), 1.46-1.38 (m, 4H), 0.77 (q, J=7.5 Hz, 6H). 13C NMR (CD3CN) δ (ppm): 175.6, 166.4, 144.8, 134.7, 134.0, 133.4, 132.3, 128.0, 127.7, 127.5, 127.2, 126.1, 125.7, 122.5, 59.5, 50.1, 40.8, 37.8, 34.3, 25.4, 11.3. HRMS m/z calculated for C23H30N5O3 [M+H]+ 424.2349, found 424.2359.
(R)-3-[4-(acetylamino-methyl)-[1,2,3]triazol-1-yl]-N-hydroxy-4-naphthalen-2-yl-butyramide (18). Compound 18 was obtained according to general procedure C from azide 110 and N-(prop-2-yn-1-yl)acetamide followed by general procedure D as a white solid (0.035 g, 14%). Purity: 100%, LC tR=1.99 min (method A), MS (ESI+): m/z=368 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.22 (s, 1H), 7.85-7.77 (m, 3H), 7.49-7.45 (m, 4H), 7.20 (dd, J=1.5 and 8.4 Hz, 1H), 6.90 (s, 1H), 5.29-5.19 (m, 1H), 4.24 (d, J=5.7 Hz, 2H), 3.40-3.37 (m, 2H), 2.82-2.77 (m, 2H), 1.81 (s, 3H). 13C NMR (CD3CN) δ (ppm): 170.4, 166.7, 144.6, 134.6, 133.3, 132.3, 128.0, 127.7, 127.5, 127.3, 126.2, 125.8, 122.7, 59.6, 40.9, 37.6, 34.5, 21.9. HRMS m/z calculated for C19H22N5O3 [M+H]+ 368.1723, found 368.1737.
(R)-3-{4-[(4-fluoro-phenylamino)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (19). Compound 9 was obtained according to general procedure C from azide 110 and 4-fluoro-N-(prop-2-yn-1-yl)aniline followed by general procedure D as a white solid (0.010 g, 8%). Purity: 100%, LC tR=2.52 min (method A), MS (ESI+): m/z=420 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.78-7.75 (m, 1H), 7.67-7.61 (m, 2H), 7.48-7.41 (m, 4H), 7.06 (d, J=8.1 Hz, 1H), 6.71 (t, J=8.7 Hz, 1.8H), 6.48-6.43 (m, 1.6H), 5.27-5.23 (m, 1H), 4.22 (s, 1.4H), 3.43-3.36 (m, 2H), 3.05 (m, 2H). 13C NMR (MeOD-d4) δ (ppm): 134.0, 133.4, 132.5, 127.8, 127.4, 127.2, 126.5, 125.8, 125.4, 115.0, 114.7, 113.7, 60.1, 48.9, 41.0, 37.5. HRMS m/z calculated for C23H23N5O2F [M+H]+ 420.1836, found 420.1828.
Tert-butyl ((1-(R)-4-(hydroxyamino)-1-(naphthalen-2-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-L-leucinate (20). Compound 20 was obtained according to general procedure C from azide 110 and tert-butyl prop-2-yn-1-yl-L-leucinate followed by general procedure D as a white solid (0.017 g, 11%). Purity: 97%, LC tR=2.67 min (method A), MS (ESI+): m/z=496 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.79-7.65 (m, 4H), 7.50-7.41 (m, 3H), 7.21 (d, J=8.1 Hz, 1H), 5.38-5.28 (m, 1H), 3.83 (dd, J=27.6 and 13.8 Hz, 2H), 3.45 (d, J=7.2 Hz, 2H), 2.97-2.84 (m, 2H), 1.70-1.29 (m, 13H), 0.88 (dd, J=10.2 and 6.3 Hz, 6H). 13C NMR (MeOD-d4) δ (ppm): 172.5, 166.9, 142.5, 134.0, 133.5, 132.5, 127.9, 127.5, 127.2, 126.6, 125.8, 125.4, 124.1, 82.2, 60.2, 59.1, 41.3, 40.9, 40.8, 37.6, 26.9, 24.7, 21.7, 21.0. HRMS m/z calculated for C27H38N5O4 [M+H]+ 496.2924, found 496.2941.
(R)—N-hydroxy-4-naphthalen-2-yl-3-{4-[(3-phenyl-ureido)-methyl]-[1,2,3]triazol-1-yl}-butyramide (21). Compound 21 was obtained according to general procedure C from azide 110 and 1-phenyl-3-(prop-2-yn-1-yl)urea followed by general procedure D as a white solid (0.051 g, 38%). Purity: 100%, LC tR=2.34 min (method A), MS (ESI+): m/z=445 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (s, 0.8H), 8.80 (s, 0.8H), 8.52 (s, 1H), 7.88 (s, 1H), 7.75-7.72 (m, 3H), 7.50-7.13 (m, 9H), 6.90 (t, J=7.2 Hz, 1H), 6.49 (t, J=5.4 Hz, 1H), 5.32-5.23 (m, 1H), 4.24 (d, J=5.4 Hz, 2H), 3.33 (m, 2H+H2O), 2.82-2.63 (m, 2H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 155.4, 145.4, 140.9, 135.1, 133.4, 132.3, 129.5, 129.1, 128.8, 128.3, 127.9, 127.7, 126.5, 126.1, 122.7, 121.6, 118.1, 59.2, 41.4, 37.7, 35.2. HRMS m/z calculated for C24H25N6O3 [M+H]+ 445.1988, found 445.1966.
(R)-[1-(2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-carbamic acid tert-butyl ester (22). Compound 22 was obtained according to general procedure C from azide 110 and tert-butyl prop-2-yn-1-ylcarbamate followed by general procedure D as an off-white oil (0.021 g, 13%). Purity: 100%, LC tR=2.44 min (method A), MS (ESI+): m/z=426 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.07 (s, 1H), 7.85-7.75 (m, 3H), 7.49-7.46 (m, 4H), 7.19 (d, J=8.4 Hz, 1H), 7.03 (s, 0.6H), 5.62 (s, 0.8H), 5.31-5.21 (m, 1H), 4.16 (d, J=6.0 Hz, 2H), 3.40-3.37 (m, 2H), 2.84-2.81 (m, 2H), 2.81 (m, 9H). 13C NMR (CD3CN) δ (ppm): 134.6, 133.4, 132.3, 128.0, 127.7, 127.5, 127.2, 126.2, 125.8, 122.1, 59.4, 41.0, 37.6, 35.7, 27.6. HRMS m/z calculated for C22H28N5O4 [M+H]+ 426.2141, found 426.2144.
(R)-3-(4-benzyloxymethyl-[1,2,3]triazol-1-yl)-N-hydroxy-4-naphthalen-2-yl-butyramide (23). Compound 23 was obtained according to general procedure C from azide 110 and ((prop-2-yn-1-yloxy)methyl)benzene followed by general procedure D as a white solid (0.066 g, 27%). Purity: 100%, LC tR=2.57 min (method A), MS (ESI+): m/z=417 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.22 (s, 1H), 7.81-7.70 (m, 3H), 7.49-7.41 (m, 4H), 7.435-7.29 (m, 3H), 7.21 (m, 3H), 5.33-5.23 (m, 1H), 4.48 (s, 2H), 4.33 (dd, J=12.0 and 14.7 Hz, 2H), 3.44-3.33 (m, 2H), 2.94-2.79 (m, 2H). 13C NMR (CD3CN) δ (ppm): 166.6, 143.7, 138.3, 134.6, 133.3, 132.3, 128.3, 128.0, 127.8, 127.7, 127.5, 127.2, 126.2, 125.8, 123.6, 71.3, 62.8, 59.5, 41.0, 37.6. HRMS m/z calculated for C24H25N4O3 [M+H]+ 417.1927, found 417.1932.
(R)-4-Fluoro-N-{2-[1-(1-hydroxycarbamoyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]triazol-4-yl]-ethyl}-benzamide (29). Compound 29 was obtained according to general procedure C from azide 130 and N-(but-3-yn-1-yl)-4-fluorobenzamide followed by general procedure D as a white solid (70 mg, 3%), Purity: 97%, LC tR=2.73 min, MS (ESI+): m/z=448 [M+1]+. 1H NMR (DMSO) δ (ppm): 8.64 (br s, 1H), 8.22 (s, 1H), 7.95-7.90 (m, 2H), 7.83-780 (m, 1H), 7.75-7.69 (m, 2H), 7.63 (s, 1H), 7.46-7.44 (m, 2H), 7.33-7.26 (m, 3H), 5.42 (t, J=7.9 Hz, 1H), 3.52-3.49 (m, 2H), 2.89-2.84 (m, 2H). 13C NMR (DMSO) δ (ppm): 165.6, 164.5, 164.3 (JC-F=247 Hz), 144.7, 134.3, 133.3, 132.4, 131.4, 130.3 (JC-F=9 Hz), 129.5, 128.8, 128.2, 127.9, 127.8, 126.5, 126.1, 122.0, 115.6 (JC-F=22 Hz), 62.3, 39.5, 38.1, 25.9. HRMS m/z calculated for C24H23N5O3F [M+H]+ 448.1785, found 448.1793.
(S)-4-fluoro-N-[1-(2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-benzamide (30) Compound 30 was obtained according to general procedure C from azide 111 and 4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure D as a white solid (0.255 g, 46%). Purity: 96%, LC tR=2.75 min, MS (ESI+): m/z=448 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (s, 0.9H), 8.99 (br t, J=5.7 Hz, 1H), 8.79 (br s, 0.9H), 7.93-7.88 (m, 3H), 7.78-7.70 (m, 3H), 7.47-7.40 (m, 3H), 7.30 (t, J=8.7 Hz, 2H), 7.14 (d, J=8.4 Hz, 1H), 5.31-5.22 (m, 1H), 4.42 (d, J=5.7 Hz, 2H), 3.32-3.28 (m, H2O+2H), 2.78 (dd, J=8.7 and 15.0 Hz, 1H), 2.66 (dd, J=5.4 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.4, 164.3 (d, JC-F=246 Hz), 144.9, 135.0, 133.3, 132.3, 131.1, 130.4 (2C), 130.3, 128.2, 127.8 (2C), 127.7, 126.5, 126.0, 123.0, 115.7 (d, JC-F=21 Hz, 2C), 59.2, 41.4, 37.6, 35.2. Mp=190° C. HRMS m/z calculated for C24H23N5O3F [M+H]+ 448.1785, found 448.1761.
Methyl (3R)-3-[4-[1-[(3,4-difluorobenzoyl)amino]-2-(4-hydroxyphenyl)ethyl]triazol-1-yl]-4-(1H-indol-3-yl)butanoate (145). The benzylated compound, obtained according to general procedure C from azide 131 and N-(1-(4-(benzyloxy)phenyl)but-3-yn-2-yl)-3,4-difluorobenzamide, (85 mg, 0.13 mmol) was put in solution in CH2Cl2 and cooled down to −78° C. before a 1.0 M solution of boron tribromide in CH2Cl2 (485 μL, 0.39 mmol, 3.0 equiv) was added dropwise. The resulting mixture was further stirred for 1 h before the reaction was quenched with iced water. The mixture was further diluted with water and layers were separated. The aqueous layer was further extracted twice with CH2Cl2. The collected organic phases were dried over MgSO4, filtered and concentrated under reduced pressure to give the desired compound. The residue was purified by flash chromatography on silica gel (CH2Cl2/MeOH 10/0 to 7/3) to afford the desired compound 145 as an off-white solid (72 mg, 98%). LC tR=2.60 min, MS (ESI+): m/z=560 [M+H]+. 1H NMR (CDCl3+1% TMS), mixture of diastereoisomers, δ (ppm): 8.49 (dd, J=13.6 and 1.7 Hz, 1H), 8.43 (br s, 0.5H), 8.31 (br s, 0.5H), 7.70-7.61 (m, 1H), 7.54-7.39 (m, 3H), 7.25-7.22 (m, 1H), 7.15-6.99 (m, 3H), 6.95 (s, 0.5H), 6.81 (s, 0.5H), 6.72 (d, J=8.4 Hz, 1H), 6.66-6.59 (m, 3H), 6.45 (dd, J=10.6 and 2.3 Hz, 1H), 5.46-5.34 (m, 1H), 5.14-4.95 (m, 1H), 3.58 (s, 3H), 3.34-3.21 (m, 2H), 3.18-2.88 (m, 4H). 13C NMR (CDCl3+1% TMS), mixture of diastereoisomers, δ (ppm): 170.7, 170.6, 164.9, 164.9, 155.2, 155.2, 152.6 (dd, J=154.8 and 12.6 Hz), 150.1 (dd, J=249.7 and 13.2 Hz), 145.9, 145.4, 136.1, 136.1, 130.8 (dd, J=6.6 and 4.2 Hz), 130.6, 130.6, 127.9, 127.8, 126.9, 126.8, 123.8 (dd, J=6.6 and 3.6 Hz), 123.3, 123.3, 122.9 (br), 122.2, 119.7, 118.1, 118.8, 117.4 (d, J=18.0 Hz), 116.9 (d, J=18.6 Hz), 115.5, 115.4, 111.5, 109.7, 109.6, 59.2, 59.0, 52.1, 47.7, 47.7, 40.7, 40.7, 38.9, 38.8, 31.5, 31.2.
3,4-Difluoro-N-[1-[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]-2-(4-hydroxyphenyl)ethyl]benzamide (146). Compound 146 was obtained from ester 145 according to general procedure G as a white solid (44 mg, 61%). Purity 94%, LC tR=2.25 min, MS (ESI+): m/z=561 [M+H]+. 1H NMR (MeOD-d4), mixture of diastereoisomers, δ (ppm): 7.63-7.55 (m, 1H), 7.55-7.48 (m, 1H), 7.45 (s, 0.5H), 7.39 (s, 0.5H), 7.38-7.34 (m, 1H), 7.33-7.24 (m, 2H), 7.07-7.00 (m, 1H), 6.98-6.91 (m, 3H), 6.78 (d, J=6.7 Hz, 1H), 6.66-6.61 (m, 2H), 5.33 (td, J=13.8 and 6.3 Hz, 1H), 5.27-5.16 (m, 1H), 3.39 (dd, J=14.4 and 5.4 Hz, 1H), 3.36-3.27 (m, 1H), 3.08-2.82 (m, 4H). 13C NMR (MeOD-d4), mixture of diastereoisomers, δ (ppm): 168.8, 167.2, 157.1, 153.6 (dd, J=251.8 and 12.7 Hz), 151.2 (dd, J=247.5 and 13.3 Hz), 148.7, 148.6, 137.9, 137.9, 137.2 (br), 133.0-132.9 (m), 131.3 (2C), 129.7, 129.7, 128.5, 128.4, 125.6-125.4 (m), 124.6, 124.6, 124.1, 124.5, 120.0, 118.9, 118.4 (d, J=18.0 Hz), 117.9 (d, J=19.0 Hz), 116.1 (2C), 112.3, 110.6, 61.2, 61.1, 49.6, 49.6, 40.9, 40.8, 38.7 (br), 32.5. HRMS m/z calculated for C29H26F2N6O4 [M+H]+ 561.2062, found 561.2062.
(R)-4-(3-bromo-phenyl)-3-{4-[(4-fluoro-benzoylamino)-methyl]-[1,2,3]triazol-1-yl}-butyric acid (128). Compound 128 was obtained according to general procedure C from azide 124 and 4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure P as a white solid (366 mg, 50%). Purity: 85%, LC tR=2.52 min, MS (ESI+): m/z=459/461 [M+H]+. 1H NMR (CDCl3) δ (ppm): 7.77-7.70 (m, 2H), 7.56-7.53 (m, 1H), 7.28-7.26 (m, 1H+CDCl3), 7.13 (s, 1H), 7.05 (t, J=7.5 Hz, 3H), 6.89 (d, J=8.4 Hz, 1H), 5.07-5.02 (m, 1H), 4.53-4.48 (m, 2H), 3.23-3.16 (m, 3H), 3.01-2.94 (m, 1H). 13C NMR (CDCl3) δ (ppm): 172.5, 166.6, 164.9 (d, JC-F=250 Hz), 138.2, 131.9, 130.4 (d, JC-F=11 Hz), 129.7, 129.6, 127.6, 124.0, 122.7, 115.6 (d, JC-F=22 Hz), 59.9, 41.0, 38.8, 34.6.
(R)-4-(4-bromo-phenyl)-3-{4-[(4-fluoro-benzoylamino)-methyl]-[1,2,3]triazol-1-yl}-butyric acid (129). Compound 129 was obtained according to general procedure C from azide 125 and 4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure P as a white solid (3.56 g, 33%). Purity: 100%, LC tR=2.82 min, MS (ESI+): m/z=463/461 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 12.38 (br s, 0.9H), 9.01 (br s, 1H), 7.96-7.91 (m, 3H), 7.35-7.27 (m, 4H), 6.94 (d, J=8.1 Hz, 2H), 5.12-5.02 (m, 1H), 4.44 (d, J=5.7 Hz, 2H), 3.12-2.85 (m, 4H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.4, 162.7, 136.8, 131.7, 131.6, 131.1, 130.4, 130.3, 123.0, 120.3, 115.8, 115.5 (d, JC-F=22 Hz), 35.3.
(R)—N-{1-[1-(4-bromo-benzyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (31). Compound 31 was obtained according to general procedure D from carboxylic acid 129 as a white solid (0.044 g, 17%). Purity: 100%, LC tR=2.76 min, MS (ESI+): m/z=478/476 [M+H]+. 1H NMR (acetone-d6+MeOD-d4) δ(ppm): 7.90-7.86 (m, 2H), 7.63 (s, 1H), 7.28 (d, J=8.1 Hz, 2H), 7.20 (t, J=8.7 Hz, 2H), 6.89 (d, J=8.1 Hz, 2H), 5.21-5.12 (m, 1H), 4.54 (s, 2H), 3.22-3.19 (m, 2H), 2.90 (dd, J=8.1 and 14.7 Hz, 1H), 2.80 (dd, J=5.4 and 14.7 Hz, 1H). 13C NMR (acetone-d6+MeOD-d4) δ (ppm): 167.4, 167.0, 164.9 (d, JC-F=249 Hz), 144.4, 135.8, 131.2, 130.6, 129.7, 129.6, 123.4, 120.4, 115.0 (d, JC-F=22 Hz), 59.8, 40.2, 37.4, 34.6. HRMS m/z calculated for C20H20N5O3BrF [M+H]+ 476.0734, found 476.0739.
(R)—N-[1-(1-biphenyl-4-ylmethyl-2-hydroxycarbamoyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-4-fluoro-benzamide (32). Compound 32 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.044 g, 43%). Purity: 100%, LC tR=2.85 min, MS (ESI+): m/z=474 [M+H]+. 1H NMR (dmso-d6) δ (ppm): 10.51 (br s, 0.9H), 9.01 (br t, J=5.7 Hz, 1H), 8.79 (br s, 0.9H), 7.93-7.89 (m, 2H), 7.84 (s, 1H), 7.53-7.21 (m, 9H), 7.03 (d, J=8.1 Hz, 2H), 5.23-5.14 (m, 1H), 4.44 (d, J=5.7 Hz, 2H), 3.16 (d, J=7.2 Hz, 2H), 2.75 (dd, J=8.7 and 15.6 Hz, 1H), 2.80 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 164.1 (d, JC-F=204 Hz), 144.9, 140.2, 138.8, 136.6, 131.1, 130.4 (d, JC-F=9 Hz), 130.0, 129.3, 127.8, 126.9, 123.2, 115.7 (d, JC-F=22 Hz), 59.8, 40.2, 37.5, 35.3. HRMS m/z calculated for C26H25N5O3F [M+H]+ 474.1941, found 474.1970.
(R)-4-fluoro-N-{1-[2-hydroxycarbamoyl-1-(4′-hydroxymethyl-biphenyl-4-ylmethyl)-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-benzamide (33). Compound 33 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.017 g, 4%). Purity: 100%, LC tR=2.57 min, MS (ESI+): m/z=504 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 9.01 (br t, J=5.7 Hz, 1H), 8.79 (br s, 0.8H), 7.94-7.89 (m, 2H), 7.86 (s, 1H), 7.49 (d, J=8.4 Hz, 2H), 7.43 (d, J=8.1 Hz, 2H), 7.35 (d, J=8.1 Hz, 2H), 7.25 (t, J=8.7 Hz, 2H), 7.03 (d, J=8.4 Hz, 2H), 5.21-5.17 (m, 2H), 4.52 (d, J=5.7 Hz, 2H), 4.44 (d, J=5.7 Hz, 2H), 3.16 (d, J=7.2 Hz, 2H), 2.75 (dd, J=8.7 and 15.0 Hz, 1H), 2.65 (dd, J=5.4 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ(ppm): 166.0, 165.4, 144.9, 142.1, 138.7, 138.5, 136.4, 131.0, 130.3 (d, JC-F=9 Hz), 130.0, 127.4, 126.8, 126.6, 123.1, 115.6 (d, JC-F=22 Hz), 63.0, 59.2, 40.2, 37.5, 35.3. HRMS m/z calculated for C27H27N5O4F [M+H]+ 504.2047, found 504.2039.
(R)-4′-(2-{4-[(4-fluoro-benzoylamino)-methyl]-[1,2,3]triazol-1-yl}-3-hydroxycarbamoyl-propyl)-biphenyl-4-carboxylic acid methylamide (34). Compound 34 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.130 g, 53%). Purity: 97%, LC tR=2.48 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.54 (br s, 0.7H), 9.03 (br t, J=5.4 Hz, 1H), 8.47 (d, J=4.2 Hz, 1H), 8.20 (br s, 1H), 7.94-7.86 (m, 5H), 7.62 (d, J=8.1 Hz, 2H), 7.50 (d, J=8.1 Hz, 2H), 7.24 (t, J=8.7 Hz, 2H), 7.06 (d, J=7.8 Hz, 2H), 5.24-5.15 (m, 1H), 4.45 (d, J=5.4 Hz, 2H), 3.18 (d, J=6.9 Hz, 2H), 2.80-2.61 (m, 4H), 2.65 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.7, 165.9, 165.4, 163.5 (d, JC-F=113 Hz), 145.0, 142.5, 137.8, 137.3, 133.6, 131.1, 130.4 (d, JC-F=9 Hz), 130.1, 128.1, 127.1, 126.7, 123.1, 115.6 (d, JC-F=22 Hz), 59.2, 40.2, 39.1, 35.3, 26.7. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2144.
(R)—N-{1-[1-(4′-acetylamino-biphenyl-4-ylmethyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (35). Compound 35 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a pale brown solid (0.132 g, 48%). Purity: 100%, LC tR=2.52 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (br s, 0.8H), 10.00 (br s, 1H), 9.01 (t, J=5.7 Hz, 1H), 8.17 (br s, 0.7H), 7.94-7.89 (m, 2H), 7.85 (s, 1H), 7.62 (d, J=8.7 Hz, 2H), 7.47 (d, J=8.7 Hz, 2H), 7.41 (d, J=8.4 Hz, 2H), 7.25 (t, J=8.7 Hz, 2H), 7.01 (d, J=8.4 Hz, 2H), 5.22-5.13 (m, 1H), 4.43 (d, J=5.7 Hz, 2H), 3.15 (d, J=8.4 Hz, 2H), 2.75 (dd, J=8.7 and 15.0 Hz, 1H), 2.64 (dd, J=5.4 and 15.0 Hz, 1H). 13C NMR
(DMSO-d6) δ (ppm): 168.8, 166.0, 165.4, 144.9, 139.2, 138.4, 136.1, 134.6, 131.1, 130.4, 130.3, 130.0, 127.1, 126.4, 119.7, 115.7 (d, JC-F=21 Hz), 59.3, 40.2, 37.5, 35.3, 24.5. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2165.
(R)—N-(1-{1-[4′-(acetylamino-methyl)-biphenyl-4-ylmethyl]-2-hydroxycarbamoyl-ethyl}-1H-[1,2,3]triazol-4-ylmethyl)-4-fluoro-benzamide (36). Compound 36 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a pale brown solid (0.130 g, 41%). Purity: 95%, LC tR=2.54 min, MS (ESI+): m/z=545 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (br s, 0.8H), 9.01 (br s, 1H), 8.35 (s, 0.9H), 8.20 (br s, 1H), 7.94-7.86 (m, 3H), 7.48-7.41 (m, 4H), 7.29-7.22 (m, 4H), 7.03 (d, J=7.2 Hz, 2H), 5.24-5.14 (m, 1H), 4.45 (d, J=3.9 Hz, 2H), 4.27 (d, J=4.5 Hz, 2H), 3.16 (d, J=6.3 Hz, 2H), 2.75 (dd, J=8.4 and 14.4 Hz, 1H), 2.67 (dd, J=4.8 and 14.4 Hz, 1H), 1.88 (s, 3H). 13C NMR (DMSO-d6) δ (ppm): 169.7, 166.0, 165.4 162.7, 145.0, 139.2, 138.7, 138.6, 136.5, 131.1, 130.3 (d, JC-F=9 Hz), 130.0, 128.3, 126.8, 123.1, 115.6 (d, JC-F=22 Hz), 59.2, 42.3, 40.9, 37.5, 35.3, 23.0. HRMS m/z calculated for C29H30N6O4F [M+H]+ 545.2313, found 545.2298.
(R)-4-fluoro-N-{1-[2-hydroxycarbamoyl-1-(3′-hydroxymethyl-biphenyl-4-ylmethyl)-ethyl]-1H-[1,2,3]-triazol-4-ylmethyl}-benzamide (37). Compound 37 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.025 g, 8%). Purity: 100%, LC tR=2.50 min, MS (ESI+): m/z=504 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (br s, 0.7H), 9.03 (br t, J=5.7 Hz, 1H), 8.80 (br s, 0.8H), 7.94-7.89 (m, 2H), 7.86 (s, 1H), 7.51 (s, 1H), 7.45-7.21 (m, 7H), 7.04 (d, J=8.1 Hz, 2H), 5.23-5.19 (m, 2H), 4.54 (d, J=5.7 Hz, 2H), 4.44 (d, J=5.7 Hz, 2H), 3.16 (d, J=7.2 Hz, 2H), 2.75 (dd, J=9.0 and 15.3 Hz, 1H), 2.64 (dd, J=5.7 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.4, 162.7, 144.9, 143.6, 139.9, 139.0, 136.5, 131.1, 131.0, 130.4 (d, JC-F=22 Hz), 130.0, 129.1, 126.9, 125.9, 125.2, 125.0, 123.1, 115.6 (d, JC-F=21 Hz), 63.3, 59.2, 41.2, 37.5, 35.3. HRMS m/z calculated for C27H27N5O4F [M+H]+ 504.2047, found 504.2042.
(R)-4′-(2-{4-[(4-fluoro-benzoylamino)-methyl]-[1,2,3]triazol-1-yl}-3-hydroxycarbamoyl-propyl)-biphenyl-3-carboxylic acid methylamide (38). Compound 38 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.130 g, 25%). Purity: 93%, LC tR=2.58 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (br s, 0.8H), 9.03 (br t, J=5.4 Hz, 1H), 8.53 (br m, 1H), 8.15 (s, 1H), 8.03 (br s, 1H), 7.93-7.86 (m, 3H), 7.79 (d, J=7.8 Hz, 1H), 7.67 (d, J=7.8 Hz, 1H), 7.53-7.49 (m, 3H), 7.23 (t, J=8.7 Hz, 2H), 7.08 (d, J=8.1 Hz, 2H), 5.25-5.16 (m, 1H), 4.44 (d, J=5.7 Hz, 2H), 3.19 (d, J=6.9 Hz, 2H), 2.80 (d, J=4.5 Hz, 3H), 2.76 (dd, J=8.7 and 15.3 Hz, 1H), 2.65 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.9, 166.0, 165.4, 163.7, 145.0, 140.2, 138.2, 137.0, 135.6, 131.1, 130.4, 130.3, 130.1, 129.4, 127.1, 126.6, 125.5, 123.1, 115.7 (d, JC-F=22 Hz), 59.2, 40.2, 37.5, 35.3, 26.7. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2151.
(R)—N-{1-[1-(3′-acetylamino-biphenyl-4-ylmethyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (39). Compound 39 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.156 g, 62%). Purity: 100%, LC tR=2.63 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.53 (br s, 0.9H), 10.00 (br s, 1H), 9.03 (br t, J=5.7 Hz, 1H), 8.81 (br s, 0.8H), 7.93-7.84 (m, 4H), 7.51 (d, J=7.8 Hz, 1H), 7.40-7.16 (m, 6H), 7.04 (d, J=7.8 Hz, 2H), 5.25-5.15 (m, 1H), 4.45 (d, J=5.7 Hz, 2H), 3.17 (d, J=6.6 Hz, 2H), 2.77 (dd, J=8.4 and 15.0 Hz, 1H), 2.65 (dd, J=5.1 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 168.9, 166.0, 165.5, 162.7, 145.0, 140.7, 140.3, 138.9, 136.7, 131.1, 130.3 (d, JC-F=9 Hz), 130.1, 129.7, 126.9, 123.1, 121.6, 118.4, 117.5, 115.6 (d, JC-F=22 Hz), 59.2, 40.2, 37.5, 35.3, 24.5. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2137.
(R)-Formic acid 4′-(2-{4-[(4-fluoro-benzoylamino)-methyl]-[1,2,3]triazol-1-yl}-3-hydroxycarbamoyl-propyl)-biphenyl-3-ylmethyl ester (40). Compound 40 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a white solid (0.026 g, 7%). Purity: 100%, LC tR=2.75 min, MS (ESI+): m/z=532 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.51 (br s, 0.7H), 9.01 (br t, J=5.7 Hz, 1H), 8.80 (br s, 0.8H), 8.34 (s, 1H), 7.93-7.88 (m, 2H), 7.85 (s, 1H), 7.58 (s, 1H), 7.49-7.34 (m, 5H), 7.23 (t, J=9.0 Hz, 2H), 7.05 (d, J=8.1 Hz, 2H), 5.21-5.14 (m, 3H), 4.44 (d, J=5.7 Hz, 2H), 3.16 (d, J=7.2 Hz, 2H), 2.76 (dd, J=8.1 and 15.6 Hz, 1H), 2.64 (dd, J=5.4 and 15.6 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.4, 162.7, 162.5, 144.9, 140.4, 138.4, 136.8, 136.8, 131.0, 130.3 (d, JC-F=9 Hz), 130.0, 129.5, 127.5, 127.0, 126.8, 126.7, 123.1, 115.6 (d, JC-F=22 Hz), 65.3, 59.2, 40.2, 37.5, 35.3. HRMS m/z calculated for C27H27N5O4F [M−COH+H]+ 504.2047, found 504.2056.
(R)-4-fluoro-N-{1-[2-hydroxycarbamoyl-1-(4-pyrimidin-5-yl-benzyl)-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-benzamide (41). Compound 17 was obtained according to general procedure Q from bromo-compound 129 followed by general procedure D as a yellow oil (0.109 g, 42%). Purity: 100%, LC tR=2.30 min, MS (ESI+): m/z=476 [1\4+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.54 (br s, 0.8H), 9.15 (br s, 1H), 9.02 (s, 3H), 7.92-7.87 (m, 2H), 7.83 (s, 1H), 7.60 (d, J=8.1 Hz, 2H), 7.23 (t, J=9.0 Hz, 2H), 7.13 (d, J=8.1 Hz, 2H), 5.27-5.17 (m, 1H), 4.44 (d, J=6.0 Hz, 2H), 3.21 (d, J=5.7 Hz, 2H), 2.79 (dd, J=9.0 and 15.3 Hz, 1H), 2.66 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 165.4, 164.2 (d, JC-F=252 Hz), 163.5, 157.6, 154.9, 145.0, 138.3, 133.2, 132.4, 131.0, 130.4, 130.3 (d, JC-F=9 Hz), 127.2, 123.2, 115.6 (d, JC-F=22 Hz), 59.1, 40.2, 37.5, 35.2. HRMS m/z calculated for C24H23N7O3F [M+H]+ 476.1846, found 476.1846.
4-{[1-(2-Hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-carbamoyl}-piperidine-1-carboxylic acid tert-butyl ester (42). Compound 42 was obtained according to general procedure C from azide 107 and tert-butyl 4-(prop-2-yn-1-ylcarbamoyl)piperidine-1-carboxylate followed by general procedure E as a white foam (130 mg, 18%). Purity: 100%, LC tR=2.73 min, MS (ESI+): m/z=439 [M+H]+. 1H-NMR (DMSO-d6) δ (ppm): 2.14 (d, 2H, J=6.4 Hz), 2.82-2.96 (m, 2H), 3.71-3.85 (m, 2H), 4.14-4.25 (m, 1H), 6.22 (d, 1H, J=8.3 Hz), 6.30 (t, 1H, J=5.0 Hz), 7.13 (t, 2H, J=8.8 Hz), 7.38 (d, 1H, J=8.0 Hz), 7.42-7.49 (m, 2H), 7.58-7.62 (m, 2H), 7.69 (br s, 1H), 7.81-7.87 (m, 3H), 8.79 (s, 1H), 9.95 (s, 1H), 10.42 (s, 1H). 13C-NMR (DMSO-d6) δ(ppm): 169.3, 167.7, 158.4 (d, JC-F=238.1 Hz), 157.9, 137.0, 135.8, 133.5, 132.2, 128.5, 128.0, 127.9, 126.4, 125.8, 121.2 (d, JC-F=7.6 Hz), 115.8 (d, JC-F=22.0 Hz), 48.9, 43.8, 37.7.
(R)-4-Fluoro-N-[1-(1-hydroxycarbamoyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-benzamide (43). Compound 43 was obtained according to general procedure C from azide 130 and 4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure D as a white solid (65 mg, 7%). Purity: 96%, LC tR=2.77 min, MS (ESI+): m/z=434 [M+1]+. 1H NMR (DMSO) δ (ppm): 9.08 (t, J=5.0 Hz, 1H), 8.22 (s, 1H), 8.00-7.95 (m, 2H), 7.85-7.77 (m, 2H), 7.73-7.70 (m, 1H), 7.64 (s, 1H), 7.47-7.40 (m, 2H), 7.38-7.29 (m, 3H), 5.45 (t, J=8.0 Hz-1H), 4.49 (d, J=5.4 Hz, 2H), 3.54-3.52 (m, 2H). 13C NMR (DMSO) δ (ppm): 165.5, 164.4 (JC-F=247 Hz), 164.3, 145.5, 134.2, 133.3, 132.4, 131.1 (JC-F=3 Hz), 130.4 (JC-F=9 Hz), 128.3, 128.0, 127.9, 127.7, 126.5, 126.2, 122.5, 115.7 (JC-F=22 Hz), 62.3, 38.0, 35.3. HRMS m/z calculated for C23H21N5O3F [M+H]+ 434.1628, found 448.1588.
2,4-Difluoro-N-[1-(2-hydroxycarbamoyl-1-naphthalen-2-ylmethyl-ethyl)-1H-[1,2,3]-triazol-4-ylmethyl]-benzamide (46). Compound 17 was obtained according to general procedure C from azide 107 and 2,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure E as a white foam (87 mg, 54%). Purity: 100%, LC tR=2.73 min, MS (ESI+): m/z=4439 [M+H]+. 1H-NMR (DMSO-d6) δ (ppm): 2.14 (d, 2H, J=6.4 Hz), 2.82-2.96 (m, 2H), 3.71-3.85 (m, 2H), 4.14-4.25 (m, 1H), 6.22 (d, 1H, J=8.3 Hz), 6.30 (t, 1H, J=5.0 Hz), 7.13 (t, 2H, J=8.8 Hz), 7.38 (d, 1H, J=8.0 Hz), 7.42-7.49 (m, 2H), 7.58-7.62 (m, 2H), 7.69 (br s, 1H), 7.81-7.87 (m, 3H), 8.79 (s, 1H), 9.95 (s, 1H), 10.42 (s, 1H). 13C-NMR (DMSO-d6) δ (ppm): 169.3, 167.7, 158.4 (d, JC-F=238.1 Hz), 157.9, 137.0, 135.8, 133.5, 132.2, 128.5, 128.0, 127.9, 126.4, 125.8, 121.2 (d, JC-F=7.6 Hz), 115.8 (d, JC-F=22.0 Hz), 48.9, 43.8, 37.7. HRMS m/z calculated for C24H21F2N5O3 [M+H]+ 466.1691, found 466.1692.
(R)—N-{1-[1-(3-bromo-benzyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (47). Compound 47 was obtained according to general procedure D from carboxylic acid 128 as a colorless oil (0.042 g, 30%). Purity: 90%, LC tR=2.38 min, MS (ESI+): m/z=478/476 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (br s, 0.9H), 9.03 (br t, J=5.7 Hz, 1H), 8.80 (br s, 0.8H), 7.97-7.92 (m, 2H), 7.86 (s, 1H), 7.34-7.25 (m, 4H), 7.12 (t, J=7.5 Hz, 1H), 6.96 (d, J=7.5 Hz, 1H), 5.22-5.13 (m, 1H), 4.45 (d, J=5.7 Hz, 2H), 3.15-3.12 (m, 2H), 2.75 (dd, J=9.3 and 15.3 Hz, 1H), 2.60 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (acetone-d6) δ (ppm): 166.4, 165.7, 164.5 (d, JC-F=248 Hz), 144.5, 139.8, 132.0, 130.9, 130.2, 129.9, (d, JC-F=9 Hz), 129.7, 128.0, 123.2, 121.9, 115.1 (d, JC-F=22 Hz), 59.4, 40.5, 37.4, 34.8. HRMS m/z calculated for C20H20N5O3BrF [M+H]+ 476.0734, found 476.0732.
(R)—N-[1-(1-biphenyl-3-ylmethyl-2-hydroxycarbamoyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-4-fluoro-benzamide (48). Compound 48 was obtained according to general procedure Q from bromo-compound 128 followed by general procedure D as a colorless oil (0.034 g, 24%). Purity: 96%, LC tR=2.53 min, MS (ESI+): m/z=474 [M+H]+. 1H NMR (acetone-d6) δ (ppm): 8.14 (s, 1H), 7.97-7.76 (m, 3H), 7.54-7.15 (m, 9H), 7.04 (d, J=7.5 Hz, 1H), 5.35-5.31 (m, 1H), 4.60-4.46 (m, 2H), 3.34 (d, J=9.6 Hz, 2H), 3.02-2.83 (m, 2H). 13C NMR (acetone-d6) δ (ppm): 166.5, 165.6, 164.5 (d, JC-F=248 Hz), 144.4, 141.0, 140.7, 137.7, 130.8, 129.9 (d, JC-F=9 Hz), 128.9, 128.8, 128.1, 127.7, 127.3, 126.8, 125.3, 123.3, 115.1 (d, JC-F=22 Hz), 59.7, 41.0, 37.5, 34.9. HRMS m/z calculated for C26H25N5O3F [M+H]+ 474.1941, found 474.1934.
(R)—N-{1-[1-(4′-acetylamino-biphenyl-3-ylmethyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (49). Compound 49 was obtained according to general procedure Q from bromo-compound 128 followed by general procedure D as a colorless oil (0.060 g, 38%). Purity: 95%, LC tR=2.15 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (acetone-d6) δ (ppm): 9.36 (br s, 0.5H), 8.46 (br s, 0.5H), 8.14 (s, 1H), 7.96-7.14 (m, 11H), 6.99 (d, J=7.2 Hz, 1H), 5.33-5.30 (m, 1H), 4.55-4.49 (m, 2H), 3.33-3.31 (m, 2H), 3.05-2.82 (m, 2H), 2.11 (s, 3H). 13C NMR (acetone-d6) δ(ppm): 168.4, 166.6, 165.7, 164.5 (d, JC-F=248 Hz), 144.4, 140.5, 138.9, 137.6, 135.4, 129.9 (d, JC-F=9 Hz), 128.9, 127.6, 127.2, 127.0, 124.9, 123.3, 119.5, 119.4, 115.1 (d, JC-F=22 Hz), 59.7, 41.1, 37.4, 34.9, 23.4. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2167.
(R)-4-fluoro-N-{1-[2-hydroxycarbamoyl-1-(4′-hydroxymethyl-biphenyl-3-ylmethyl)-ethyl]-1H-[1,2,3]-triazol-4-ylmethyl}-benzamide (50). Compound 50 was obtained according to general procedure Q from bromo-compound 128 followed by general procedure D as a white solid (0.013 g, 22%). Purity: 97%, LC tR=2.18 min, MS (ESI+): m/z=504 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.53 (br s, 0.9H), 9.03 (br t, J=5.7 Hz, 1H), 8.80 (br s, 0.9H), 7.95-7.89 (m, 3H), 7.51-7.23 (m, 9H), 6.96 (d, J=7.8 Hz, 1H), 5.24-5.20 (m, 2H), 4.52 (d, J=5.7 Hz, 2H), 4.45 (d, J=5.7 Hz, 2H), 3.20 (d, J=7.2 Hz, 2H), 2.78 (dd, J=8.7 and 15.3 Hz, 1H), 2.65 (dd, J=5.4 and 15.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.4, 164.1 (d, JC-F=204 Hz), 144.9, 142.2, 140.5, 138.8, 138.0, 131.1, 130.4 (d, JC-F=9 Hz), 129.3, 128.3, 127.6, 127.4, 126.8, 125.4, 123.2, 115.7 (d, JC-F=22 Hz), 63.0, 59.2, 41.3, 37.4, 35.3. HRMS m/z calculated for C27H27N5O4F [M+H]+ 504.2047, found 504.2033.
(R)—N-{1-[1-(3′-acetylamino-biphenyl-3-ylmethyl)-2-hydroxycarbamoyl-ethyl]-1H-[1,2,3]triazol-4-ylmethyl}-4-fluoro-benzamide (51). Compound 51 was obtained according to general procedure Q from bromo-compound 128 followed by general procedure D as a colorless oil (0.069 g, 41%). Purity: 100%, LC tR=2.22 min, MS (ESI+): m/z=531 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.53 (br s, 0.8H), 10.04 (br s, 1H), 9.03 (br t, J=5.7 Hz, 1H), 7.94-7.89 (m, 3H), 7.79 (s, 1H), 7.56 (d, J=7.8 Hz, 1H), 7.37-7.18 (m, 7H), 6.95 (d, J=7.8 Hz, 1H), 5.27-5.17 (m, 0.9H), 4.45 (d, J=5.7 Hz, 2H), 3.20 (d, J=6.9 Hz, 2H), 2.78 (dd, J=8.7 and 15.0 Hz, 1H), 2.65 (dd, J=5.7 and 15.0 Hz, 0.9H). 13C NMR (DMSO-d6) δ (ppm): 168.9, 165.5, 164.3 (d, JC-F=245 Hz), 145.0, 141.0, 140.7, 140.2, 138.1, 131.1, 131.0, 130.4 (d, JC-F=9 Hz), 129.7, 129.4, 128.6, 127.7, 125.4, 123.1, 121.9, 118.6, 117.7, 115.7 (d, JC-F=22 Hz), 59.3, 41.3, 37.5, 35.3, 24.5. HRMS m/z calculated for C28H28N6O4F [M+H]+ 531.2156, found 531.2172.
(R)-4-fluoro-N-{1-[2-hydroxycarbamoyl-1-(3′-hydroxymethyl-biphenyl-3-ylmethyl)-ethyl]-1H-[1,2,3]-triazol-4-ylmethyl}-benzamide (52). Compound 52 was obtained according to general procedure Q from bromo-compound 128 followed by general procedure D as a white solid (0.040 g, 32%). Purity: 100%, LC tR=2.27 min, MS (ESI+): m/z=504 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.53 (br s, 0.9H), 9.02 (br t, J=5.7 Hz, 1H), 8.81 (br s, 0.8H), 7.94-7.88 (m, 3H), 7.50-7.23 (m, 9H), 6.96 (d, J=7.8 Hz, 1H), 5.28-5.20 (m, 2H), 4.56 (d, J=6.3 Hz, 2H), 4.46 (d, J=5.7 Hz, 2H), 3.21 (d, J=6.3 Hz, 2H), 2.79 (dd, J=9.0 and 15.0 Hz, 1H), 2.65 (dd, J=5.4 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.5, 164.1 (d, JC-F=206 Hz), 144.9, 143.6, 140.8, 140.3, 138.1, 131.1, 131.0, 130.4 (d, JC-F=9 Hz), 129.3, 129.1, 128.5, 127.8, 126.0, 125.5, 125.2, 123.1, 115.6 (d, JC-F=22 Hz), 63.4, 59.2, 41.3, 37.4, 35.3. HRMS m/z calculated for C27H27N5O4F [M+H]+ 504.2047, found 504.2043.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (54). Compound 54 was obtained according to general procedure C from azide 107 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure E as a white foam (62 mg, 45%), Purity: 100%, LC tR=2.47 min, MS (ESI+): m/z=466 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.56 (s, 1H), 7.71-7.60 (m, 4H), 7.70-7.51 (m, 1H), 7.47 (s, 1H), 7.45 (s, 1H), 7.42-7.25 (m, 2H), 7.25-7.19 (m, 1H), 7.14-7.10 (dd, J=1.8 Hz, J=10.2 Hz, 1H), 5.35-5.22 (m, 1H), 4.46 (s, 2H), 3.39 (d, J=7.5 Hz, 2H), 3.31 (dd, J=1.5 Hz, J=3.3 Hz, 1H), 2.85 (dd, J=1.5 Hz, J=3.3 Hz, 1H). 13C NMR (MeOD) δ (ppm): 168.6, 167.4, 145.5, 135.4, 134.0, 133.0, 129.2, 128.8, 128.7, 128.5 (2C), 128.0, 127.1, 126.7, 125.5, 125.0, 118.6, 118.4, 118.0, 117.8, 61.5, 42.5, 38.9, 36.0. 19F NMR (MeOD) δ (ppm): −136.3 (d, J=19.7 Hz), −140.0 (d, J=19.7 Hz). HRMS m/z calculated for C24H22FN5O3 [M+H]+ 466.1691, found 466.1688.
4-[(dimethylamino)methyl]-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (55). Compound 55 was obtained according to general procedure C from azide 107 and 4-((dimethylamino)methyl)-N-(prop-2-yn-1-yl)benzamide followed by general procedure E as a white solid (13 mg, 16%). Purity: 100%, LC tR=1.88 min, MS (ESI+): m/z=487 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.49 (s, 1H), 7.80 (d, J=8.1 Hz, 2H), 7.71-7.63 (m, 4H), 7.51 (d, J=8.1 Hz, 2H), 7.44 (s, 1H), 7.39-7.32 (m, 2H), 7.12 (dd, J=1.5 Hz, J=8.4 Hz, 1H), 5.32-5.28 (m, 1H), 4.50 (s, 2H), 4.03 (s, 2H), 3.42-3.39 (m, 2H), 2.95-2.84 (m, 2H), 2.62 (s, 6H). 13C NMR (MeOD) δ (ppm): 169.2, 168.5, 145.6, 137.9, 135.9, 135.4, 134.8, 134.0, 133.9, 131.5 (2C), 129.2, 129.0 (2C), 128.8, 128.6, 128.5, 128.0, 127.1, 126.7, 124.8, 62.7, 61.5, 43.9, 42.4, 38.8, 36.1. HRMS m/z calculated for C27H31N6O3 [M+H]+ 487.2458, found 487.2458.
4-fluoro-N-[2-[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide) (56). Compound 56 was obtained according to general procedure C from azide 107 and N-(but-3-yn-1-yl)-4-fluorobenzamide followed by general procedure F as a white foam (27 mg, 54%), Purity: 100%, LC tR=2.10 min, MS (ESI+): m/z=462 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.55 (s, 1H), 7.79-7.71 (m, 3H), 7.67 (d, J=8.1 Hz, 2H), 7.52 (s, 1H, H5), 7.44-7.36 (m, 3H), 7.17-7.11 (m, 3H), 5.29-5.27 (m, 1H), 3.50 (t, J=4.8 Hz, 2H), 3.39 (t, J=4.8 Hz, 2H), 2.98-2.81 (m, 4H, H2). 13C NMR (MeOD) δ (ppm): 167.7, 167.2, 166.4, 163.1, 144.2, 134.1, 133.4, 132.5, 130.5, 129.5, 127.8, 127.4 (2C), 127.1, 126.6, 125.8, 125.4, 122.8, 115.1, 114.8, 60.0, 41.0, 39.3, 37.6, 24.9. 19F NMR (MeOD) δ (ppm): −111.2. HRMS m/z calculated for C25H25FN5O3 [M+H]+ 462.1942, found 462.1953.
3,4-difluoro-N-[2-[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide (57). Compound 57 was obtained according to general procedure C from azide 107 and N-(but-3-yn-1-yl)-3,4-difluorobenzamide followed by general procedure F as a white foam (27 mg, 89%). Purity: 100%, LC tR=2.50 min, MS (ESI+): m/z=480 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.54 (s, 1H), 7.74-7.64 (m, 4H), 7.59-7.40 (m, 1H), 7.39 (s, 1H), 7.39-7.14 (m, 4H), 7.11 (dd, J=1.5 Hz, J=8.4 Hz, 1H), 5.36-5.16 (m, 1H), 3.52-3.41 (td, J=3.3 Hz, J=7.2 Hz, 2H), 3.35 (t, J=7.2 Hz, 2H), 2.93-2.83 (m, 4H, H2). 13C NMR (MeOD) δ (ppm): 167.2, 166.3, 154.0, 151.6, 150.7, 148.3, 144.1, 134.1, 133.4, 132.5, 131.5, 127.8, 127.2, 126.6, 125.8, 125.4, 124.1, 122.9, 117.2, 116.5, 60.0, 41.0, 39.4, 37.6, 24.9. 19F NMR (MeOD) δ(ppm): −136.5 (d, J=17.8 Hz), −140.0 (d, J=20.6 Hz). HRMS m/z calculated for C25H24F2N5O3 [M+H]+ 480.1847, found 480.1856.
3-{4-[(4-Fluoro-phenylcarbamoyl)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (58). Compound 58 was obtained according to general procedure C from azide 107 and N-(4-fluorophenyl)but-3-ynamide followed by general procedure G as a white foam (3 mg, 1.2% over 2 steps). Purity: 100%, LC tR=2.47 min, MS (ESI+): m/z=448 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.86 (dd, J=5.7 Hz, J=14.9 Hz, 1H), 2.96 (dd, J=8.9 Hz, J=14.9 Hz, 1H), 3.40-3.43 (m, 2H), 4.74 (s, 2H), 3.68 (s, 2H), 5.25-5.35 (m, 1H), 7.03 (t, J=8.8 Hz, 2H), 7.35-7.39 (m, 2H), 7.45-7.51 (m, 3H), 7.65-7.74 (m, 4H). 13C NMR (MeOD) δ (ppm): 34.5, 38.9, 42.4, 61.6, 116.2 (d, J=22.9 Hz, 2C), 123.1 (d, J=8.0 Hz, 2C), 125.1, 126.7, 127.1, 128.0, 128.5, 128.6, 128.9, 129.2, 133.9, 134.8, 135.4, 135.9, 142.0, 160.7 (d, J=241.5 Hz), 168.6, 169.9. 19F NMR (MeOD) δ (ppm): −120.9. HRMS m/z calculated for C24H23FN5O3 [M+H]+ 448.1785, found 448.1809.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-3,4-dimethoxy-benzamide (60). Compound 17 was obtained according to general procedure C from azide 107 and 3,4-dimethoxy-N-(prop-2-yn-1-yl)benzamide followed by general procedure F as a white solid (25 mg, 66%). Purity: 100%, LC tR=3.22 min, MS (ESI+): m/z=444 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.93-7.79 (m, 1H), 7.80-7.60 (m, 3H), 7.43-7.29 (m, 5H), 7.12 (d, J=7.2 Hz, 1H), 6.99 (d, J=7.2 Hz, 1H), 5.38-5.26 (m, 1H), 4.55-4.42 (m, 2H), 3.83 (s, 3H), 3.84 (s, 3H), 3.50-3.37 (m, 2H), 3.20 (dd, J=14.7 and 7.5 Hz, 1H), 2.81-3.02 (m, 1H). HRMS m/z calculated for C24H23FN5O3 [M+H]+ 448.1785, found 448.1801.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-1,3-benzodioxole-5-carboxamide (61). Compound 61 was obtained according to general procedure C from azide 107 and N-(prop-2-yn-1-yl)benzo[d][1,3]dioxole-5-carboxamide followed by general procedure F as a white solid (22 mg, 45%). Purity: 100%, LC tR=2.65 min, MS (ESI+): m/z=474 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.67-7.59 (m, 4H), 7.42-7.30 (m, 4H), 7.21 (d, J=1.8 Hz, 1H), 7.12 (dd, J=8.1 and 1.8 Hz, 1H), 6.85 (d, J=8.1 Hz, 1H), 6.04 (s, 2H), 5.31-5.26 (m, 1H), 4.46 (s, 2H), 3.43-3.37 (m, 2H), 2.95 (dd, J=15.0 and 8.7 Hz, 1H), 2.86 (dd, J=15.0 and 5.7 Hz, 1H). HRMS m/z calculated for C25H23N5O5 [M+H]+ 474.1777 found 474.1772.
3-chloro-4-fluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (62). Compound 62 was obtained according to general procedure C from azide 107 and 3-chloro-4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure F as a white solid (25 mg, 47%). Purity: 100%, LC tR=2.87 min, MS (ESI+): m/z=482 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.52 (br s, 1H), 9.11 (t, J=5.7 Hz, 1H), 8.80 (br s, 1H), 8.05 (dd, J=4.8 and 2.1 Hz, 1H), 7.90-7.85 (m, 2H), 7.78-7.67 (m, 3H), 7.54 (t, J=8.7 Hz, 1H), 7.46-7.35 (m, 3H), 7.13 (dd, J=8.4 and 1.5 Hz, 1H), 5.32-5.22 (m, 1H), 4.42 (d, J=5.7 Hz, 2H), 3.34-3.26 (m, 2H+DMSO), 2.78 (dd, J=15.0 and 8.7 Hz, 1H), 2.67 (dd, J=15.0 and 5.7 Hz, 1H). 19F NMR (MeOD-d4) δ (ppm): −113.7. HRMS m/z calculated for C24H21ClFN5O3 [M+H]+ 482.1395, found 482.1393.
2,3,4-Trifluoro-N-[1-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]-triazol-4-ylmethyl]-benzamide (63). Compound 63 was obtained according to general procedure C from azide 107 and 2,3,4-trifluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (15 mg, 10% over 2 steps). Purity:
100%, LC tR=2.50 min, MS (ESI+): m/z=484 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.88 (dd, J=5.7 Hz, J=14.9 Hz, 1H), 2.97 (dd, J=9.0 Hz, J=14.9 Hz, 1H), 3.38-3.45 (m, 2H), 4.48 (br s, 1H), 5.25-5.34 (m, 1H), 7.58 (s, 1H), 7.64-7.71 (m, 3H). 13C NMR (MeOD) δ (ppm): 36.0, 38.9, 42.4, 61.5, 113.7 (dd, J=18.6 Hz, 2C), 118.5 (d, J=18.0 Hz), 124.8, 125.5 (dd, J=3.8, J=7.3 Hz), 126.3 (dd, J=3.5, J=7.2 Hz), 125.6 (d, J=3.7 Hz, J=18.1 Hz), 122.0 (d, J=11.0 Hz), 124.9, 130.8 (2C), 125.8 (br s), 126.7, 127.0, 128.0, 128.5 (2C), 128.53, 128.8, 129.2, 133.9, 134.8, 135.4, 141.0 (td, J=15.9 Hz, J=250.3 Hz), 150.7 (ddd, J=3.3 Hz, J=11.2 Hz, J=254.4 Hz), 154.0 (ddd, J=2.9 Hz, J=9.9 Hz, J=253.5 Hz), 164.6, 168.5. 19F NMR (MeOD) δ (ppm): −133.6 (dd, J=9.7 Hz, J=20.0 Hz), −137.6 (dd, J=9.7 Hz, J=20.0 Hz), −163.3 (t, J=20.0 Hz). HRMS m/z calculated for C24H21F3N5O3 [M+H]+ 484.1596, found 484.1644.
Methyl (3R)-3-[4-[[(3,4-difluorobenzoyl)amino]methyl]triazol-1-yl]-4-(1-naphthyl)butanoate (64) Compound 64 was obtained according to general procedure C from azide 147 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (45 mg, 58% over 2 steps). Purity: 95%, LC tR=2.45 min, MS (ESI+): m/z=466 [M+H]+. 1H NMR (MeOD) δ (ppm) (ppm): 2.98-3.01 (m, 2H), 3.53 (dd, J=10.2 Hz, J=14.0 Hz, 1H), 3.82 (dd, J=4.5 Hz, J=14.0 Hz, 1H) 4.43 (d, J=2.5 Hz, 2H), 5.38-5.28 (m, 1H), 6.94 (d, J=7.0 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 7.32-7.54 (m, 4H), 7.59-7.73 (m, 3H), 7.77 (d, J=8.0 Hz, 1H), 8.05 (d, J=8.0 Hz, 1H). 13C NMR (MeOD) δ (ppm): 35.9, 38.9, 39.5, 60.8, 117.9 (d, J=19.0 Hz), 118.4 (d, J=17.9 Hz), 124.9, 124.9, 125.6 (dd, J=3.7 Hz, J=7.3 Hz), 126.2, 126.8, 127.5, 128.6, 128.9, 129.9, 132.6, 133.3, 133.8, 135.3, 145.4, 150.9 (dd, J=13.3 Hz; J=176.0 Hz), 154.2 (dd, J=12.9 Hz; J=186.8 Hz), 167.4, 168.6. 19F NMR (MeOD) δ (ppm): −136.5 (d, J=17.8 Hz), −140.0 (d, J=20.6 Hz). HRMS m/z calculated for C24H22F2N5O3 [M+H]+ 466.1691, found 466.1718.
N-[[1-[(1R)-1-[(4-chlorophenyl)methyl]-3-(hydroxyamino)-3-oxo-propyl]triazol-4-yl]methyl]-3,4-difluoro-benzamide (65). Compound 65 was obtained according to general procedure C from azide 148 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (55 mg, 28% over 4 steps). Purity: 100%, LC tR=2.40 min, MS (ESI+): m/z=450 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.81 (dd, J=5.7 Hz, J=14.8 Hz, 2H), 2.90 (dd, J=8.9 Hz, J=14.8 Hz, 2H), 3.19-3.26 (m, 2H), 4.52 (s, 2H), 5.10-5.20 (m, 1H), 6.94 (d, J=8.3 Hz, 2H), 7.12 (d, J=8.3 Hz, 2H), 7.37 (td, J=8.3 Hz, J=10.2 Hz, 1H), 7.61 (s, 1H), 7.65-7.69 (m, 1H), 7.75 (ddd, J=2.1 Hz, J=7.7 Hz, J=11.1 Hz, 1H). 13C NMR (MeOD) δ (ppm): 36.1, 38.7, 41.5, 61.3, 117.9 (d, J=18.8 Hz), 118.5 (d, J=18.0 Hz), 124.9, 125.5 (dd, J=3.7 Hz, J=7.3 Hz), 129.6 (2C), 131.6 (2C), 132.6, 133.8, 136.7, 145.6, 151.4 (dd, J=13.1 Hz, J=248.4 Hz), 153.8 (dd, J=12.6 Hz, J=252.2 Hz), 167.5, 168.4. 19F NMR (MeOD) δ (ppm): −136.3 (d, J=20.5 Hz), −140.0 (d, J=20.5 Hz). HRMS m/z calculated for C20H19C1F2N5O3 [M+H]+ 450.1144, found 450.1172.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]pyrimidine-4-carboxamide (66). Compound 65 was obtained according to general procedure C from azide 107 and N-(prop-2-yn-1-yl)pyrimidine-4-carboxamide followed by general procedure G. White foam (40 mg, 20% over 2 steps). Purity: 100%, LC tR=2.05 min, MS (ESI+): m/z=432 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.86 (dd, J=5.6 Hz, J=14.9 Hz), 2.97 (dd, J=9.6 Hz, J=14.9 Hz), 3.39-3.35 (m, 2H), 4.53 (s, 2H), 5.33-5.23 (m, 1H), 7.11 (dd, J=1.5 Hz, J=8.4 Hz, 1H), 7.32-7.27 (m, 2H), 7.37 (s, 1H), 7.65-7.56 (m, 4H), 7.97 (dd, J=1.1 Hz, J=5.1 Hz, 1H), 8.98 (d, J=5.1 Hz, 1H), 9.21 (d, J=1.1 Hz, 1H). 13C NMR (MeOD) δ (ppm): 35.5, 38.8, 42.5, 61.5, 119.8, 125.0, 126.7, 127.0, 128.0, 128.4, 128.5, 128.8, 129.2, 133.8, 134.8, 135.3, 145.1, 157.7, 159.0, 160.4, 164.7, 168.5. HRMS m/z calculated for C22H21N7O3 [M+H]+ 432.1784, found 432.1783.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-4-(2-methoxyethoxy)benzamide (68). Compound 68 was obtained according to general procedure C from azide 107 and 4-(2-methoxyethoxy)-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (75 mg, 42% over 2 steps). Purity: 100%, LC tR=2.33 min, MS (ESI+): m/z=504 [M+H]+. 1H NMR (MeOD) δ(ppm): 2.97-2.81 (m, 2H), 3.39-7.30 (m, 2H), 3.42 (s, 3H), 3.75 (td, J=3.1 Hz, J=6.2 Hz, 2H), 4.16 (td, J=3.1 Hz, J=6.2 Hz, 2H), 4.47 (s, 1H), 5.33-5.24 (m, 1H), 6.97 (d, J=8.9 Hz, 2H), 7.11 (dd, J=1.4 Hz, J=8.4 Hz, 1H), 7.36-7.32 (m, 2H), 7.41 (s, 1H), 7.61 (s, 1H), 7.73-7.62 (m, 5H). 13C NMR (MeOD) δ (ppm): 35.9, 38.9, 42.4, 59.3, 61.5, 68.6, 72.0, 115.3 (2C), 124.7, 126.7, 127.1, 127.5, 128.0, 128.5 (2C), 128.8, 129.2, 130.3 (2C), 133.8, 134.8, 135.3, 146.0, 163.1, 168.5, 169.4. HRMS m/z calculated for C27H30N5O5 [M+H]+504.2247, found 504.2224.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-1-methyl-imidazole-4-carboxamide (69). Compound 69 was obtained according to general procedure C from azide 107 and 1-methyl-N-(prop-2-yn-1-yl)-1H-imidazole-4-carboxamide followed by general procedure G as a white foam (23.2 mg, 29% over 2 steps). Purity: 100%, LC tR=2.00 min, MS (ESI+): m/z=434 [M+H]+. 1H NMR (MeOD) δ: 2.83 (dd, J=5.7 Hz, J=14.8 Hz, 1H), 2.93 (dd, J=8.9 Hz, J=14.8 Hz, 1H), 3.37-3.40 (m, 2H), 3.34 (s, 3H), 4.78 (s, 2H), 5.23-5.33 (m, 1H), 7.11 (dd, J=1.7 Hz, J=8.4 Hz, 1H), 7.32-7.38 (m, 2H), 7.42 (s, 1H), 7.57 (d, J=6.7 Hz, 2H), 7.56-7.71 (m, 4H). 13C NMR (MeOD) δ (ppm): 34.0, 35.1, 38.9, 42.4, 61.5, 124.7, 125.1, 126.7, 127.0, 128.0, 128.5, 128.6, 128.8, 129.2, 133.9, 134.8, 135.4, 137.1, 139.8, 145.7, 164.8, 168.5. HRMS m/z calculated for C22H24N7O3 [M+H]+ 434.1941, found 434.1946.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-3-oxo-1-(2-phenylethyl)propyl]triazol-4-yl]methyl]benzamide (70). Compound 70 was obtained according to general procedure C from azide 149 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (41 mg, 18% over 4 steps). Purity: 100%, LC tR=2.37 min, MS (ESI+): m/z=430 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.15-2.39 (m, 2H), 2.41-2.49 (m, 2H), 4.63 (s, 2H), 4.90-4.98 (m, 1H), 7.08-7.25 (m, 5H), 7.36 (ddd, J=8.0 Hz, J=8.5 Hz, J=10.3 Hz, 1H), 7.72 (dddd, J=1.4 Hz, J=2.2 Hz, J=4.3 Hz, J=8.5 Hz, 1H), 7.80 (ddd, J=2.2 Hz, J=7.7 Hz, J=11.3 Hz, 1H), 7.87 (s, 1H). 13C NMR (MeOD) δ (ppm): 32.8, 36.3, 37.5, 39.7, 59.5, 117.9 (d, J=19.0 Hz), 118.5 (d, J=17.9 Hz), 124.5, 125.6, 125.6 (dd, J=3.7, J=7.3 Hz), 127.2, 129.4 (2C), 129.5 (2C), 132.7, 141.7, 145.8, 154.8 (dd, J=13.0 Hz, J=248.0 Hz), 155.7 (dd, J=12.7 Hz, J=252.0 Hz), 167.7, 168.5. 19F NMR (MeOD) δ (ppm): −136.2 (d, J=20.5 Hz), −140.0 (d, J=20.5 Hz). HRMS m/z calculated for C21H22F2N5O3 [M+1-1]+ 430.1691, found 430.1670.
Tert-butyl 3-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methylcarbamoyl]morpholine-4-carboxylate (71). Compound 71 was obtained according to general procedure C from azide 107 and tert-butyl 3-(prop-2-yn-1-ylcarbamoyl)morpholine-4-carboxylate followed by general procedure G as a white foam (59 mg, 17% over 2 steps). Purity: 100%, LC tR=2.43 min, MS (ESI+): m/z=539 [M+H]+. 1H NMR (MeOD) δ (ppm): 1.41 (s, 9H3 (Boc)), 2.80-3.94 (m, 4H), 3.38-3.43 (m, 3H), 3.54-3.60 (m, 1H), 3.65-3.79 (m, 2H), 4.11-4.21 (m, 1H), 4.29-4.38 (m, 3H), 5.28-5.32 (m, 1H), 7.13-7.17 (2×t, J=2.0 Hz, 1H), 7.40-7.43 (m, 2H), 7.50 and 7.58 (2×brs, 2H), 7.71-7.79 (m, 3H). 13C NMR (MeOD) δ (ppm): 28.5 (3C), 35.7, 39.1, 42.3, 61.4 (2C), 61.5, 67.4 (2C), 68.7, 82.0, 124.5, 126.8, 127.2, 128.0, 128.6 (2C), 128.8, 129.3, 133.9, 134.9, 135.4 (Cdia), 135.5 (Cdia), 145.8, 168.5, 172.3.
2-chloro-4-fluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (72). Compound 72 was obtained according to general procedure C from azide 107 and 2-chloro-4-fluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (28 mg, 22% over 2 steps). Purity: 100%, LC tR=2.43 min, MS (ESI+): m/z=482 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.87 (dd, J=5.7 Hz, J=14.9 Hz, 1H), 2.96 (dd, J=8.8 Hz, J=14.9 Hz, 1H), 3.41-3.44 (m, 2H), 4.48 (d, J=1.9 Hz, 1H), 5.28-5.37 (m, 1H), 7.08 (td, J=2.5 Hz, J=8.4 Hz, 1H), 7.14 (dd, J=1.7 Hz, J=8.4 Hz, 1H), 7.24 (dd, J=2.5 Hz, J=8.8 Hz, 1H), 7.30 (dd, J=6.0 Hz, J=8.6 Hz, 1H), 7.38-7.41 (m, 2H), 7.48 (s, 1H), 7.64 (s, 1H), 7.68-7.77 (m, 3H). 13C NMR (MeOD) δ (ppm): 35.9, 39.0, 42.3, 61.5, 115.3 (d, J=21.8 Hz), 118.3 (d, J=25.3 Hz), 124.9, 126.8, 127.1, 128.0, 128.6 (2C), 128.8, 129.9, 131.7, 131.8, 133.5 (dd, J=3.4 Hz, J=7.6 Hz), 133.9, 134.9, 135.4, 145.2, 164.4 (d, J=251 Hz), 168.5, 168.9. 19F NMR (MeOD) δ (ppm): −111.3. HRMS m/z calculated for C24H22FC1N5O3 [M+H]+482.1395, found 482.1436.
4-hydroxy-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide4-hydroxy-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (73). Compound 73 was obtained according to general procedure C from azide 107 and 4-hydroxy-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (16 mg, 9% over 2 steps). Purity: 100%, LC tR=2.10 min, MS (ESI+): m/z=446 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.84 (dd, J=5.7 Hz, J=14.9 Hz, 1H), 2.94 (dd, J=8.9 Hz, J=14.9 Hz, 1H), 3.43-3.37 (m, 2H), 4.74 (s, 2H), 5.33-5.24 (m, 1H), 6.80 (d, J=8.8 Hz, 2H), 7.11 (dd, J=1.7 Hz, J=8.4 Hz, 1H), 7.40-7.31 (m, 2H), 7.42 (s, 1H), 7.70-7.60 (m, 6H). 13C NMR (MeOD) δ (ppm): 35.9, 38.9, 42.4, 61.5, 116.1 (2C), 124.7, 126.0, 126.7, 127.1, 128.0, 128.5 (2C), 128.8, 129.2, 130.4 (2C), 133.8, 134.8, 135.3, 146.1, 162.2, 168.5, 169.7. HRMS m/z calculated for C24H23N5O4 [M+H]+ 446.1828, found 446.1832.
3-{4-[(3,4-Difluoro-phenylcarbamoyl)-methyl]-[1,2,3]triazol-1-yl}-N-hydroxy-4-naphthalen-2-yl-butyramide (74). Compound 74 was obtained according to general procedure C from azide 107 and N-(3,4-difluorophenyl)but-3-ynamide followed by general procedure G as a white foam (22 mg, 14% over 2 steps). Purity: 100%, LC tR=2.57 min, MS (ESI+): m/z=466 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.86 (dd, 1H, J=5.4 Hz, J=14.8 Hz, 1H), 2.97 (dd, 1H, J=8.9 Hz; J=14.8 Hz, 1H), 3.39-3.45 (m, 2H), 3.67 (s, 2H), 5.25-5.35 (m, 1H), 7.15-7.19 (m, 3H), 7.32-7.40 (m, 2H), 7.43 (s, 1H), 7.60-7.73 (m, 5H). 13C NMR (MeOD) δ (ppm): 34.5, 38.9, 42.5, 61.5, 110.3 (d, J=21.8 Hz), 116.9 (dd, J=3.6 Hz, J=5.6 Hz), 118.1 (dd, J=1.0 Hz, J=18.3 Hz), 125.2, 126.7, 127.1, 128.0, 128.5, 128.6, 128.9, 129.2, 133.9, 134.8, 135.4, 136.8 (d, J=2.9 Hz), 141.8, 147.9 (dd, J=12.7 Hz, J=243.4 Hz), 151.1 (dd, J=12.9 Hz, J=244.5 Hz), 168.6, 169.9. 19F NMR (MeOD) δ (ppm): −139.6 (d, J=21.2 Hz), −146.5 (d, J=21.2 Hz). HRMS m/z calculated for C24H21F2N5O3 [M+H]+ 466.1691, found 466.1686.
3,4-difluoro-N-[[1-[(E,1R)-1-[2-(hydroxyamino)-2-oxo-ethyl]-4-phenyl-but-3-enyl]triazol-4-yl]methyl]benzamide (75). Compound 75 was obtained according to general procedure C from azide 139 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (117 mg, 60% over 2 steps) Purity: 100%, LC tR=2.43 min, MS (ESI+): m/z=442 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.92-2.76 (m, 4H), 4.57 (s, 2H), 5.17-5.08 (m, 1H), 6.04 (td, J=7.2 Hz, J=115.9 Hz, 1H), 6.30 (d, J=15.9 Hz, 1H), 7.22-7.10 (m, 5H), 7.32 (td, J=8.2 Hz, J=10.1 Hz, 1H), 7.63 (ddd, J=1.5 Hz, J=4.1 Hz, J=8.5 Hz, 1H), 7.70 (ddd, J=2.2 Hz, J=7.7 Hz, J=11.2 Hz, 1H), 7.90 (s, 1H). 13C NMR (MeOD) δ (ppm): 36.2, 38.7, 39.7, 59.9, 117.9 (d, J=18.8 Hz), 118.5 (d, J=18.1 Hz), 124.5, 125.0, 125.5 (dd, J=3.7, J=7.2 Hz), 127.1 (2C), 128.4, 129.4 (2C), 132.6, 135.2, 138.2, 145.7, 151.4 (dd, J=13.0 Hz, J=248 Hz), 153.8 (dd, J=13.0 Hz, J=252.0 Hz), 167.5, 168.6. 19F NMR (MeOD) δ (ppm): −136.3 (d, J=20.5 Hz), −139.9 (d, J=20.5 Hz). HRMS m/z calculated for C22H22F2N5O3 [M+H]+ 442.1691, found 442.1707.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-3-oxo-1-[[4-(trifluoromethyl)phenyl]methyl]propyl]triazol-4-yl]methyl]benzamide (76). Compound 76 was obtained according to general procedure C from azide 150 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (37 mg, 21% over 4 steps). Purity: 100%, LC tR=2.52 min, MS (ESI+): m/z=484 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.97-2.80 (m, 2H), 3.35 (d, J=7.6 Hz, 2H), 4.53 (s, 2H), 5.29-5.19 (m, 1H), 7.18 (d, J=8.0 Hz, 2H), 7.36 (dt, J=8.2 Hz, J=10.1 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.69-7.65 (m, 2H), 7.76 (ddd, J=2.0 Hz, J=7.6 Hz, J=10.3 Hz, 1H). 13C NMR (MeOD) δ (ppm): 36.1, 38.9, 41.8, 61.1, 117.9 (d, J=18.6 Hz, 2C), 118.5 (d, J=18.0 Hz), 124.8, 125.5 (dd, J=3.8, J=7.3 Hz), 126.3 (dd, J=3.5, J=7.2 Hz), 125.6 (d, J=271 Hz), 130, 130.4, 130.8 (2C), 132.7, 142.6, 145.7, 151.4 (dd, J=13.2 Hz, J=250.0 Hz), 153.8 (dd, J=12.8 Hz, J=250.0 Hz), 167.5, 168.3. 19F NMR (MeOD) δ (ppm): −64.6 (s, 3F), −136.2 (d, J=20.4 Hz), −140.0 (d, J=20.4 Hz). HRMS m/z calculated for C21H19FN7O6 [M+H]+ 484.1408, found 484.1436.
3,4-Difluoro-N-{1-[1-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]triazol-4-yl]-1-methyl-ethyl}-benzamide (77). Compound 77 was obtained according to general procedure C from azide 107 and 3,4-difluoro-N-(2-methylbut-3-yn-2-yl)benzamide followed by general procedure G as a white foam (46 mg, 37% over 2 steps). Purity: 95%, LC tR=2.62 min, MS (ESI+): m/z=494 [M+H]+. 1H NMR (MeOD) δ(ppm): 1.60 (s, 3H), 1.64 (s, 3H), 2.86 (dd, J=5.9 Hz, J=14.8 Hz, 1H), 2.94 (dd, J=8.6 Hz, J=14.8 Hz, 1H), 4.60 (br s, 1H, OH), 5.20-5.29 (m, 1H), 7.26-7.34 (m, 1H), 7.35-7.40 (m, 3H), 7.54-7.59 (m, 1H), 7.57 (s, 1H), 7.63-7.76 (m, 4H). 13C NMR (MeOD) δ (ppm): 28.2, 28.5, 38.8, 42.5, 52.5, 61.5, 118.0 (d, J=19.5 Hz), 118.3 (d, J=18.5 Hz), 123.2, 125.7 (dd, J=5.6 Hz, J=7.2 Hz), 126.7, 127.0, 128.2, 128.5, 128.6, 128.9, 129.2, 133.9, 134.8, 135.4, 151.2 (dd, J=12.9 Hz, J=248.5 Hz), 153.8, 153.5 (dd, J=12.9 Hz, J=250.0 Hz), 167.3, 168.6. 19F NMR (MeOD) δ (ppm): −136.8 (d, J=20.4 Hz), −140.2 (d, J=20.4 Hz). HRMS m/z calculated for C26H26F2N5O3 [M+H]+ 494.2004, found 494.2005.
3,4-difluoro-N-[1-[1-[3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide (78). Compound 78 was obtained according to general procedure C from azide 107 and N-(but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as a white foam (24 mg, 20% over 2 steps). Purity: 100%, LC tR=2.53 min, MS (ESI+): m/z=480 [M+H]+. 1H NMR (MeOD) δ (ppm): 1.44 and 1.48 (2d, J=7.0 Hz, J=6.9 Hz, 3H), 2.87-3.03 (m, 2H), 3.37-3.42 (m, 2H), 5.22-5.28 (m, 2H), 5.38-5.28 (m, 1H), 7.13-7.15 (m, 1H), 7.31-7.41 (m, 3H), 7.53-7.71 (m, 5H). 13C NMR (MeOD) δ(ppm): 20.3 (Cdia), 20.5 (Cdia), 38.8, 42.5, 43.4, 61.5, 117.9 (d, J=18.8 Hz, Cdia), 118.4 (d, J=17.8 Hz, Cdia), 123.8 (d, J=24.8 Hz), 125.6 (br s), 126.7, 127.1, 128.0, 128.5 (3C), 128.8, 129.2, 132.8, 133.8, 134.8, 135.4, 150.4 (d, J=12.2 Hz), 151.2 (dd, J=13.0 Hz; J=248.2 Hz), 153.7 (dd, J=13.0 Hz; J=252.0 Hz), 166.8, 168.6. 19F NMR (MeOD) δ(ppm): −136.4 (d, J=20.2 Hz), −140.0 (d, J=20.2 Hz, Fdia), −140.1 (d, J=20.2 Hz, Fdia). HRMS m/z calculated for C25H24F2N5O3 [M+H]+ 480.1837, found 480.1847.
3-[4-(2-Acetylamino-phenoxymethyl)-[1,2,3]triazol-1-yl]-N-hydroxy-4-naphthalen-2-yl-butyramide (79). Compound 79 was obtained according to general procedure C from azide 107 and N-(2-(prop-2-yn-1-yloxy)phenyl)acetamide followed by general procedure G as a white foam (39 mg, 8% over 2 steps). Purity: 100%, LC tR=2.37 min, MS (ESI+): m/z=460 [M+H]+. 1H NMR (MeOD-d4, 300 MHz) δ (ppm): 2.07 (s, 3H), 2.88 (dd, J=5.6 and 15.0 Hz, 1H), 2.97 (dd, J=8.9 and 15.0 Hz, 2H), 3.39-3.46 (m, 2H), 5.10 (s, 2H), 5.31 (dddd, J=5.6, 5.7, 8.9 and 9.0 Hz, 1H), 6.88-6.95 (m, 1H), 6.98-7.01 (m, 2H), 7.06 (dd, J=1.6 and 8.4 Hz, 1H), 7.38-7.41 (m, 2H), 7.43 (s br, 1H), 7.57-7.65 (m, 2H), 7.73-7.76 (m, 2H), 7.90 (d, J=7.9 Hz). 13C NMR (MeOD) δ (ppm): 23.8, 38.9, 42.4, 61.7, 62.8, 113.8, 122.3, 123.9, 125.9, 126.1, 126.8, 127.2, 127.9, 128.6 (2C), 128.8, 129.3, 133.9, 134.9, 135.3 (2C), 144.0, 150.0, 168.5, 171.8. HRMS m/z calculated for C25H26N5O4 [M+H]+ 460.1985, found 460.1982.
4-fluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-3-methyl-benzamide (80). Compound 80 was obtained according to general procedure C from azide 107 and 4-fluoro-3-methyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white powder (20 mg, 16% over 2 steps). Purity: 100%, LC tR=2.48 min, MS (ESI+): m/z=462 [M+H]+. 1H NMR (MeOD): δ (ppm): 2.29 (d, J=1.9 Hz, 3H), 2.90 (ddd, J=5.6, J=15.0, 2H), 3.37-3.41 (m, 2H), 4.47 (s, 2H), 5.26-5.32 (m, 1H), 7.05-7.14 (m, 2H), 7.29-7.38 (m, 2H), 7.42 (s, 1H), 7.56-7.69 (m, 6H). 13C NMR (MeOD): δ (ppm): 14.5 (d, J=3.4 Hz), 35.9, 38.9, 42.4, 61.5, 116.0 (d, J=22.5 Hz), 124.8, 126.1, 126.3, 126.7, 127.1, 128.0, 128.3 (d, J=8.0 Hz), 128.5, 128.8, 129.2, 131.3, 132.2 (d, J=8.0 Hz), 133.8, 134.8, 135.4, 145.8, 165.0 (d, J=245.9 Hz), 168.5, 168.9. 19F NMR (MeOD): δ (ppm): −115.3. HRMS m/z calculated for C25H25F2N5O3 [M+H]+ 462.1941, found 462.1980.
N-(3,4-difluorophenyl)-3-[1-[3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]propanamide (81). Compound 81 was obtained according to general procedure C from azide 107 and N-(3,4-difluorophenyl)pent-4-ynamide followed by general procedure G as a white foam (28 mg, 10% over 2 steps). Purity: 100%, LC tR=2.57 min, MS (ESI+): m/z=480 [M+H]+. 1H NMR (MeOD) δ (ppm): 2.54-2.61 (m, 2H), 2.81-2.97 (m, 4H), 3.37-3.40 (m, 2H), 5.20-5.30 (m, 1H), 7.09 (dd, J=1.6 Hz, J=8.4 Hz, 1H), 7.11-7.18 (m, 2H), 7.40-7.31 (m, 2H), 7.36-7.39 (m, 2H), 7.43 (s, 1H), 7.47 (s, 1H), 7.59-7.71 (m, 4H). 13C NMR (MeOD) δ (ppm): 1.9, 36.9, 39.0, 42.4, 61.4, 110.3 (d, J=22.0 Hz), 116.9 (dd, J=3.5 Hz, J=5.4 Hz), 118.1 d, J=18.4 Hz), 124.0, 126.7, 127.1, 127.9, 128.5 (2C), 128.8, 129.2, 133.8, 134.8, 135.4, 136.8 (dd, J=2.7 Hz, J=9.0 Hz), 147.8 (dd, J=12.6 Hz, J=246.7 Hz), 151.0 (dd, J=11.7 Hz, J=246.7 Hz), 168.6, 172.6. 19F NMR (MeOD) δ (ppm): −139.7 (d, J=21.5 Hz), −146.8 (d, J=21.5 Hz). HRMS m/z calculated for C25H24F2N5O3 [M+1-1]+ 460.1847, found 460.1877.
N-[[1-[(1S)-1-(3,4-dihydro-1H-isoquinolin-2-ylmethyl)-3-(hydroxyamino)-3-oxo-propyl]triazol-4-yl]methyl]-3,4-difluoro-benzamide (82) Compound 82 was obtained according to general procedure C from azide 122 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (14 mg, 16% over 2 steps). Purity: 100%, LC tR=2.02 min, MS (ESI+): m/z=471 [M+H]+. 1H NMR (MeOD) δ(ppm): 2.58-2.63 (m, 1H), 2.72-2.85 (m, 5H), 2.94 (dd, J=5.5 Hz, J=13.2 Hz, 1H), 2.94 (dd, J=5.4 Hz, J=13.3 Hz, 1H), 3.06 (dd, J=8.7 Hz, J=13.3 Hz, 1H), 3.53 (d, J=14.7 Hz, 1H), 3.68 (d, J=14.7 Hz, 1H), 4.59 (s, 2H), 5.21-5.31 (m, 1H), 6.92-7.08 (m, 4H), 7.31 (td, J=8.1 Hz, J=10.3 Hz, J=16.5 Hz, 1H), 7.65 (ddd, J=1.5 Hz, J=4.1 Hz, J=8.5 Hz, 1H), 7.70 (ddd, J=2.2 Hz, J=7.7 Hz, J=11.2 Hz, 1H), 7.94 (s, 1H). 13C NMR (MeOD) δ (ppm): 30.0, 36.2, 37.2, 52.1, 57.0, 57.9, 62.5, 117.9 (d, J=18.8 Hz), 118.5 (d, J=18.2 Hz), 125.0, 125.5 (dd, J=3.5 Hz, J=7.0 Hz), 126.7, 127.2, 127.4, 129.5, 132.6, 135.2, 135.5, 145.6, 151.3 (dd, J=13.7 Hz, J=248.0 Hz), 153.3 (dd, J=11.9 Hz, J=251.7 Hz), 167.5, 168.7. 19F NMR (MeOD) δ (ppm): −139.9 (d, J=20.4 Hz), −136.2 (d, J=20.4 Hz). HRMS m/z calculated for C23H25F2N6O3 [M+H]+ 471.1956 found 471.1950.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (83). Compound 83 was obtained according to general procedure C from azide 131 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (61 mg, 54% over 2 steps). Purity: 100%, LC tR=2.22 min, MS (ESI+): m/z=455 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 2.63 (dd, J=5.7 Hz, J=15.0 Hz, 1H), 2.74 (dd, J=8.5 Hz, J=15.0 Hz, 1H), 3.24 (d, J=7.1 Hz, 2H), 4.45 (d, J=5.7 Hz, 2H), 5.14-5.24 (m, 1H), 7.90 (d, J=2.3 Hz, 1H), 7.94 (td, J=1.0 Hz, J=6.9 Hz, 1H), 7.02 (td, J=1.1 Hz, J=7.0 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.57 (d, J=6.7 Hz, 1H), 7.73 (d, J=7.8 Hz, 1H), 7.56 (ddd, J=8.3 Hz, J=8.5 Hz, J=10.5 Hz, 1H), 7.74-7.79 (m, 1H), 7.91 (ddd, J=2.0 Hz, J=7.8 Hz, J=11.6 Hz, 1H), 7.92 (s, 1H), 8.79 (br s, NHOH), 9.09 (t, J=5.4 Hz, 1H, NH), 10.8 (br s, NH). 13C NMR (MeOD) δ (ppm): 32.4, 36.1, 38.7, 61.1, 110.6, 112.3, 118.0 (d, J=18.9 Hz), 118.5 (d, J=18.1 Hz), 118.9, 119.9, 122.5, 124.6, 124.7, 125.6 (dd, J=3.5 Hz, J=7.1 Hz), 128.4, 132.7, 137.9, 145.3, 151.3 (dd, J=12.5 Hz, d, J=247.4 Hz), 153.7 (dd, J=13.1 Hz, d, J=252.5 Hz), 167.5, 168.9. 19F NMR (MeOD) δ (ppm): −140.0 (d, J=20.4), −136.3 (d, J=20.4). HRMS m/z calculated for C22H21F2N6O3 [M+H]′ 455.1643, found 455.1653.
3,4-Difluoro-N-{1-[1-(1(R)-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]triazol-4-yl]-(S)-ethyl}-benzamide (85). Compound 85 was obtained according to general procedure C from azide 107 and (R)—N-(but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as a white powder (32 mg, 30% over 2 steps). Diastereoisomeric yield (dr): 92%. Purity: 100%, LC tR=2.48 min, MS (ESI+): m/z=4=480 [M+H]. 1H NMR (MeOD) δ (ppm): 1.48 (d, J=7.0 Hz, 3H), 2.87 (dd, J=5.8 Hz, J=15.0 Hz, 2H), 2.97 (dd, J=9.2 Hz, J=15.0 Hz, 2H), 3.33-3.45 (m, 2H), 5.21-5.33 (m, 2H), 7.13 (dd, J=1.6 Hz, J=8.4 Hz, 1H), 7.27-7.39 (m, 3H), 7.40 (br s, 1H), 7.56 (s, 1H), 7.57-7.70 (m, 5H). 13C NMR (MeOD) δ (ppm): 20.5, 38.8, 42.5, 43.4, 61.5, 117.9 (d, J=19.1 Hz), 118.4 (d, J=18.0 Hz), 123.6, 125.6 (dd, J=3.5, J=7.2 Hz), 126.7, 127.1, 128.0, 128.5 (2C), 128.8, 129.4, 132.8 (t, J=4.3), 133.8, 134.8, 135.4, 150.2, 151.2 (dd, J=13.0 Hz, J=247.6 Hz), 153.3 (dd, J=12.7 Hz, J=252.4 Hz), 166.8, 168.6. 19F NMR (MeOD) δ (ppm): −140.1 (d, J=20.2 Hz), −136.5 (d, J=20.2 Hz). HRMS m/z calculated for C25H24F2N5O3 [M+H]+ 480.1815, found 480.1847.
3,4-Difluoro-N-{1-[1-(1(R)-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-1H-[1,2,3]triazol-4-yl]-(R)-ethyl}-benzamide (86). Compound 86 was obtained according to general procedure C from azide 107 and (S)—N-(but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as a white powder (27 mg, 21% over 2 steps), diastereoisomeric yield (dr): 83%, Purity: 100%, LC tR=2.55 min, MS (ESI+): m/z=4=480 [M+H]. 1H NMR (MeOD) δ (ppm): 1.44 (d, J=7.0 Hz, 3H), 2.87 (dd, J=5.6 Hz, J=15.0 Hz, 2H), 2.97 (dd, J=9.0 Hz, J=15.0 Hz, 2H), 3.35-3.43 (m, 2H), 5.21-5.31 (m, 2H), 7.13 (dd, J=1.6 Hz, J=8.4 Hz, 1H), 7.26-7.34 (m, 1H), 7.36-7.40 (m, 3H), 7.53 (s, 1H), 7.63-7.72 (m, 5H). 13C NMR (MeOD) δ (ppm): 20.5, 38.8, 42.5, 43.4, 61.5, 117.9 (d, J=19.1 Hz), 118.4 (d, J=18.0 Hz), 123.6, 125.6 (dd, J=3.5, J=7.2 Hz), 126.7, 127.1, 128.0, 128.5 (2C), 128.8, 129.4, 132.8 (t, J=4.3), 133.8, 134.8, 135.4, 150.2, 151.2 (dd, J=13.0 Hz, J=247.6 Hz), 153.3 (dd, J=12.7 Hz, J=252.4 Hz), 166.8, 168.6. 19F NMR (MeOD) δ (ppm): −140.1 (d, J=20.2), −136.5 (d, J=20.2). HRMS m/z calculated for C25H24F2N5O3 [M+H]+ 480.1876, found 480.1847.
Methyl (3R)-3-[4-[[(3,4-difluorobenzoyl)-methyl-amino]methyl]triazol-1-yl]-4-(2-naphthyl)butanoate (87). Compound 87 was obtained according to general procedure C from azide 107 and 3,4-difluoro-N-methyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white powder (65 mg, 46% over 2 steps). Purity: 100%, LC tR: 2.46 min, MS (ESI+): m/z=480 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.41 (br s, 1H), 8.81 (br s, 1H), 7.81 (m, 3H), 7.47 (m, 3H), 7.27 (s, 1H), 7.15 (m, 4H), 5.28 (m, 1H), 4.29 (s, 2H), 3.37 (dd, J=5.1 and 13.8 Hz, 1H), 3.26 (m, 1H), 2.71 (m, 2H), 2.59 (s, 3H). HRMS m/z calculated for C25H24F2N5O3 [M+H]+ 480.1847 found 480.1837.
(3R)-3-[4-[(2S)-1-(3,4-difluorobenzoyl)pyrrolidin-2-yl]triazol-1-yl]-4-(1H-indol-3-yl)butanehydroxamic acid (88). Compound 88 was obtained according to general procedure C from azide 131 and (S)-(3,4-difluorophenyl)(2-ethynylpyrrolidin-1-yl)methanone followed by general procedure G as a white solid (S/R ratio 4.4/1.5 mg, 59% over 2 steps). Purity: 99%, LC tR=2.20 min, MS (ESI+): m/z=495 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.46-7.21 (m, 5H), 7.16-6.82 (m, 4H), 6.61-6.57 (m, 0.3H), 5.27-5.10 (m, 2H), 3.70-3.54 (m, 1H), 3.49-3.22 (m, 3H), 3.02-2.83 (m, 2H), 2.29-2.10 (m, 1H), 2.08-1.94 (m, 1H), 1.91-1.57 (m, 3H). 13C NMR (75 MHz, MeOD-d4) δ (ppm): 170.3, 169.5, 168.9, 168.7, 152.7 (dd, J=250.8, 12.4 Hz), 151.2 (dd, J=248.3, 12.7 Hz), 150.7, 149.7, 148.9, 145.4, 137.8, 134.9 (dd, J=8.4, 4.2 Hz), 128.5, 125.4 (dd, J=6.4, 4.3 Hz), 124.7, 124.6, 124.3, 124.1, 124.0, 122.6, 122.5, 120.0, 118.9, 118.6 (d, J=18.0 Hz), 117.9 (d, J=19.3 Hz), 117.0 (d, J=19.3 Hz), 112.4, 112.3, 110.8, 110.7, 110.5, 61.4, 61.3, 61.1, 57.4, 54.9, 51.3, 51.0, 47.4, 38.8, 38.6, 34.7, 33.2, 33.2, 32.8, 32.5, 32.2, 30.8, 30.5, 25.8, 25.7, 23.7, 22.8. HRMS m/z calculated for C25H24F2N6O3 [M+H]+ 495.1956 found 495.1957.
N-[2-(4-benzyloxyphenyl)-1-[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]-3,4-difluoro-benzamide (89). Compound 89 was obtained according to general procedure C from azide 131 and N-(1-(4-(benzyloxy)phenyl)but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as a white solid (71 mg, 63% over 2 steps). Purity: 99%, LC tR=2.80 min, MS (ESI+): m/z=651 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.65-7.56 (m, 1H), 7.55-7.48 (m, 1H), 7.43-7.24 (m, 9H), 7.07-6.92 (m, 4H), 6.81-6.74 (m, 3H), 5.41-5.31 (m, 1H), 5.27-5.15 (m, 1H), 4.96 (s, 1H), 4.95 (s, 1H), 3.42-3.28 (m, 2H), 3.11-2.82 (m, 4H). 13C NMR (75 MHz, MeOD-d4) δ(ppm): 168.8, 167.1, 158.9, 153.6 (dd, J=252.4, 12.5 Hz), 151.2 (dd, J=248.2, 13.9 Hz), 148.5, 148.3, 138.7, 137.9, 132.9 (dd, J=4.8, 3.6 Hz), 131.4, 131.1, 129.4, 128.8, 128.55, 128.49, 128.44, 125.6-125.5 (m), 124.7, 124.6, 124.19, 124.17, 122.5, 120.0, 119.0, 118.4 (d, J=17.8 Hz), 117.9 (d, J=18.6 Hz), 115.8, 112.3, 110.6, 70.9, 61.1, 49.6, 49.5, 41.0, 40.8, 38.9, 38.8, 32.5, 32.4. HRMS m/z calculated for C36H32F2N6O4 [M+H]+ 651.2531 found 651.2531.
(3R)-3-[4-[4-(3,4-difluorobenzoyl)morpholin-3-yl]triazol-1-yl]-4-(1H-indol-3-yl)butanehydroxamic acid (90). Compound 90 was obtained according to general procedure C from azide 131 and (3,4-difluorophenyl)(3-ethynylmorpholino)methanone followed by general procedure G as an off-white solid (55 mg, 62% over 2 steps). Purity: 99%, LC tR=2.15 min, MS (ESI+): m/z=511 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.53-7.26 (m, 5H), 7.14-6.77 (m, 4H), 5.32-5.22 (m, 1H), 4.03 (br s, 2H), 3.77-3.68 (m, 1H), 3.52-3.25 (m, 3H), 3.09-2.90 (m, 2H), 2.79-2.69 (m, 0.5H), 2.51 (br s, 0.5H). 13C NMR (75 MHz, MeOD-d4) δ (ppm): 170.3, 168.9, 152.6 (dd, J=252.4, 13.0 Hz), 151.4 (dd, J=248.9, 13.8 Hz), 145.5, 145.0, 144.6, 137.83, 137.80, 133.4 (dd, J=9.4, 4.8 Hz), 128.5, 126.0, 125.5-125.2 (m), 125.4, 124.7, 122.5, 120.01, 119.97, 118.9, 118.7, 118.1 (d, J=18.7 Hz), 118.0 (d, J=18.7 Hz), 112.3, 110.7, 69.5 (br s2), 67.7, 61.3, 49.3, 38.6, 38.5, 32.5. HRMS m/z calculated for C25H24F2N6O4 [M+H]+ 511.1905, found 511.1906.
3,4-difluoro-N-[1-[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide (91). Compound 91 was obtained according to general procedure C from azide 131 and N-(but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as an off-white solid (60 mg, 73% over 2 steps). Purity: 99%, LC tR=2.18 min, MS (ESI+): m/z=469 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.72 (ddd, J=11.3, 7.7, 2.2 Hz, 1H), 7.66-7.60 (m, 1H), 7.49 (d, J=3.0 Hz, 1H), 7.38-7.31 (m, 2H), 7.29-7.24 (m, 1H), 7.03 (dddd, J=8.1, 6.9, 3.4, 1.2 Hz, 1H), 6.94 (dddd, J=8.1, 6.9, 1.4, 1.2 Hz, 1H), 6.83 (d, J=3.0 Hz, 1H), 5.29-5.16 (m, 2H), 3.43-3.28 (m, 2H), 3.00-2.82 (m, 2H), 1.49 (d, J=7.1 Hz, 1.5H), 1.45 (d, J=7.1 Hz, 1.5H). 13C NMR (75 MHz, MeOD-d4) δ (ppm): 168.9, 166.9, 153.7 (dd, J=252.2, 12.6 Hz), 151.3 (dd, J=247.7, 12.6 Hz), 150.0, 137.9, 132.9 (dd, J=5.4, 3.6 Hz), 128.5, 125.7 (dd, J=7.2, 3.2 Hz), 124.7, 124.6, 123.8, 123.8, 122.5, 119.9, 118.9, 118.4 (d, J=18.0 Hz), 118.0 (d, J=18.6 Hz), 112.3, 110.7, 110.6, 61.2, 61.1, 43.5, 38.6, 32.5, 20.51, 20.46. HRMS m/z calculated for C23H22F2N6O3 [M+H]+ 469.1800, found 469.1800.
3,4-difluoro-N-[(1R)-2-hydroxy-1-[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide (92). Compound 92 was obtained according to general procedure C from azide 131 and (R)-3,4-difluoro-N-(1-hydroxybut-3-yn-2-yl)benzamide followed by general procedure G as a white solid (31 mg, 35% over 2 steps). Purity: 98%, LC tR=2.03 min, MS (ESI+): m/z=485 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.77 (br dd, J=10.8, 7.8 Hz, 1H), 7.69-7.63 (br m, 1H), 7.56 (br s, 1H), 7.38-7.22 (m, 3H), 7.03 (br t, J=7.3 Hz, 1H), 6.94 (br t, J=7.3 Hz, 1H), 6.83 (br s, 1H), 5.29 (br t, J=5.9 Hz, 1H), 5.24-5.17 (m, 1H), 3.83-3.69 (m, 2H), 3.44-3.30 (m, 2H), 2.99-2.82 (m, 2H). 13C NMR (75 MHz, MeOD-d4) δ (ppm): 168.8, 167.6, 153.7 (dd, J=252.1, 12.9 Hz), 151.2 (dd, J=247.6, 12.9 Hz), 146.5, 137.9, 132.8 (dd, J=4.2, 3.6 Hz), 128.5, 125.7 (dd, J=7.0, 3.6 Hz), 124.6, 124.5, 122.5, 119.9, 118.9, 118.4 (d, J=18.3 Hz), 118.1 (d, J=19.1 Hz), 112.3, 110.6, 64.5, 61.3, 50.2, 38.6, 32.4. HRMS m/z calculated for C23H22F2N6O4 [M+H]+ 485.1749, found 485.1746.
(3R)-3-[4-[(2S)-1-(3,4-difluorobenzoyl)pyrrolidin-2-yl]triazol-1-yl]-4-(2-naphthyl)butanehydroxamic acid (93). Compound 93 was obtained according to general procedure C from azide 107 and (S)-(3,4-difluorophenyl)(2-ethynylpyrrolidin-1-yl)methanone followed by general procedure G as a white solid (65 mg, 60% over 2 steps). Purity: 97%, LC tR=2.45 min, MS (ESI+): m/z=506 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ (ppm): 7.80-7.58 (m, 3H), 7.52-7.26 (m, 6H), 7.21-7.12 (m, 2H), 6.89-6.80 (m, 0.4H), 6.64 (brs, 0.3H), 5.30-5.20 (m, 2H), 3.68-3.19 (m, 4H), 3.06-2.87 (m, 2H), 2.27-2.08 (m, 1H), 2.05-1.88 (m, 1H), 1.76-1.58 (m, 2H). 13C NMR (75 MHz, MeOD-d4) δ(ppm): 168.8, 168.0, 167.2, 167.0, 151.3 (dd, J=250.7, 12.3 Hz), 149.8 (dd, J=248.9, 13.1 Hz), 148.4, 147.8, 147.6, 134.1, 133.6-133.3 (m), 133.4, 132.4, 127.8, 127.45, 127.37, 127.2, 126.7, 126.6, 125.9, 125.7, 125.6, 125.5, 125.4, 125.3, 124.0 (dd, J=6.3, 3.4 Hz), 123.1, 122.8, 117.2 (dd, J=18.0 Hz), 117.1 (d, J=18.0 Hz), 116.5 (d, J=18.6 Hz), 115.7 (d, J=18.6 Hz), 60.3, 60.1, 56.0, 53.5, 53.4, 50.0, 49.5, 45.9, 41.3, 41.2, 41.0, 37.6, 37.5, 37.3, 33.4, 31.9, 31.6, 31.2, 29.2, 29.0, 24.2, 21.0. HRMS m/z calculated for C27H25F2N5O3 [M+H]+ 506.2004, found 506.1997.
N-[2-(4-benzyloxyphenyl)-1-[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]-3,4-difluoro-benzamide (94). Compound 94 was obtained according to general procedure C from azide 107 and N-(1-(4-(benzyloxy)phenyl)but-3-yn-2-yl)-3,4-difluorobenzamide followed by general procedure G as a white solid (48 mg, 39% over 2 steps). Purity: 97%, LC tR=2.97 min, MS (ESI+): m/z=662 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.56 (br s, 1H), 8.86 (t, J=8.8 Hz, 1H), 8.85 (br s, 1H), 7.89 (s, 1H), 7.85-7.76 (m, 2H), 7.74-7.64 (m, 3H), 7.58-7.48 (m, 1H), 7.45-7.27 (m, 8H), 7.14-7.08 (m, 3H), 6.86-6.80 (m, 2H), 5.33-5.20 (m, 2H), 4.99 (s, 2H), 3.32-3.23 (m, 2H), 3.12-2.94 (m, 2H), 2.86-2.66 (m, 2H). 13C NMR (75 MHz, DMSO-d6) δ (ppm): 165.5, 163.3, 156.8, 151.3 (dd, J=250.0, 13.2 Hz), 149.0 (dd, J=246.6, 13.4 Hz), 147.8, 147.7, 137.2, 134.60, 134.56, 132.9, 131.8, 131.7-131.6 (br s), 130.6, 130.5, 130.1, 128.4 (2C), 127.8, 127.7 (2C), 127.4 (2C), 127.3, 126.0, 125.6, 124.9-124.7 (m), 122.1, 121.9, 117.5 (d, J=17.5 Hz), 116.7 (d, J=18.3 Hz), 114.4 (2C), 69.1, 59.0, 58.8, 47.8, 41.1, 41.0, 37.2, 37.1, 29.0. HRMS m/z calculated for C38H33F2N5O4 [M+H]+ 662.2579, found 662.2577.
(3R)-3-[4-[4-(3,4-difluorobenzoyl)morpholin-3-yl]triazol-1-yl]-4-(2-naphthyl)butanehydroxamic acid (95). Compound 95 was obtained according to general procedure C from azide 107 and (3,4-difluorophenyl)(3-ethynylmorpholino)methanone followed by general procedure G as a white solid (43 mg, 55% over 2 steps). Purity: 95%, LC tR=2.38 min, MS (ESI+): m/z=522 [M+H]+. 1H NMR (300 MHz, MeOD-d4) δ(ppm): 7.76-7.72 (m, 2H), 7.69-7.65 (m, 1H), 7.58 (d, J=11.9 Hz, 1H), 7.41-7.36 (m, 3H), 7.29-7.21 (m, 3H), 7.11 (br s, 1H), 5.37-5.27 (m, 1H), 4.23-3.99 (m, 1H), 3.75-3.23 (m, 5H), 3.13-2.90 (m, 2H), 2.66-2.56 (m, 0.5H), 2.36 (br s, 0.5H). 13C NMR (75 MHz, MeOD-d4) δ (ppm): 170.4 (br s), 168.6, 168.5, 152.6 (dd, J=250.4, 12.4 Hz), 151.4 (dd, J=248.8, 13.9 Hz), 145.1, 144.6, 135.6, 135.5, 134.8, 133.9, 133.8, 133.5-133.2 (m), 129.4, 129.3, 128.6 (2C), 128.1, 128.0, 127.21, 127.17, 126.8, 126.4, 125.5, 125.5-125.1 (m), 118.8 (d, J=18.0 Hz), 118.1 (br d, J=17.5 Hz), 118.0 (d, J=18.9 Hz), 69.4, 67.6, 67.5, 61.6, 61.4, 42.8, 42.7, 38.8, 38.7. HRMS m/z calculated for C27H25F2N5O4 [M+H]+ 522.1953, found 522.1949.
3,4-difluoro-N-[(1R)-2-hydroxy-1-[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]ethyl]benzamide (96). Compound 96 was obtained according to general procedure C from azide 107 and (R)-3,4-difluoro-N-(1-hydroxybut-3-yn-2-yl)benzamide followed by general procedure G as a white solid (39 mg, 43% over 2 steps). Purity: 95%, LC tR=2.25 min, MS (ESI+): m/z=496 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 8.75 (d, J=8.4 Hz, 1H), 7.98-7.89 (m, 1H), 7.86-7.66 (m, 4H), 7.55 (td, J=10.4, 8.4 Hz, 1H), 7.46-7.38 (m, 3H), 7.11 (dd, J=8.4, 1.4 Hz, 1H), 5.28-2.14 (m, 2H), 4.99 (br s, 1H), 3.69 (dd, J=10.5, 5.5 Hz, 1H), 3.60 (dd, J=10.5, 8.4 Hz, 1H), 3.30-3.17 (m, 2H), 2.81 (dd, J=15.0, 8.7 Hz, 1H), 2.69 (dd, J=15.0, 5.5 Hz, 1H). 13C NMR (75 MHz, DMSO-d6) δ (ppm): 165.5, 163.9, 151.3 (dd, J=250.2, 12.6 Hz), 149.0 (dd, J=246.2, 12.9 Hz), 145.8, 134.6, 132.9, 131.8 (2C), 127.8, 127.4 (3C), 127.3, 126.0, 125.6, 125.0 (dd, J=7.2, 3.0 Hz), 122.4, 117.4 (d, J=17.5 Hz), 116.9 (d, J=18.0 Hz), 63.1, 58.9, 49.1, 41.1, 37.0. HRMS m/z calculated for C25H23F2N5O4 [M+H]+ 496.1796, found 496.1795.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-N-methyl-benzamide (97). Compound 97 was obtained according to general procedure C from azide 131 and 3,4-difluoro-N-methyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a faint pink solid (22 mg, 4% over 2 steps). Purity: 90%, LC tR=2.23 min, MS (ESI+): m/z=469 [M+H]+. 1H NMR (300 MHz, CDCl3) δ (ppm): 10.0, (s, 1H), 7.50 (s, 1H), 7.37 (d, J=8.00 Hz, 2H), 7.24 (d, J=8.00 Hz, 2H), 6.96-6.82 (m, 4H), 5.19 (s, 1H) 4.54-4.29 (m, 2H), 3.31-3.28 (m, 2H) 2.70 (s, 3H), 1.36-1.18 (m, 2H). 13C NMR (75 MHz, CDCl3) δ (ppm): 166.4, 151.4, 151.2, 149.1, 148.9, 136.4, 133.9, 133.9, 127.3, 124.4, 123.6, 123.5, 124.3, 118.8, 118.1, 117.4, 116.9, 116.7, 111.3, 109.9, 68.3, 59.2, 37.5, 31.1. HRMS m/z calculated for C23H22F2N6O3 [M+H]+ 469.1800, found 469.1800.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-N-isobutyl-benzamide (98). Compound 98 was obtained according to general procedure C from azide 131 and 3,4-difluoro-N-isobutyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a faint pink solid (55 mg, 18% over 2 steps). Purity: 99%, LC tR=2.53 min, MS (ESI+): m/z=511 [M+H]+. 1H NMR (300 MHz, CDCl3) δ (ppm): 8.73, (s, 1H), 7.33-7.29, (m, 1H), 7.23-7.20 (m 2H), 7.51 (m, 1H), 7.11-6.96 (m, 4H), 5.19 (s, 1H) 4.54-4.29 (m, 2H), 3.31-3.28 (m, 2H) 2.69 (s, 3H), 1.36-1.31 (m, 1H), 1.17-1.03 (m, 1H), 0.85-0.60 (m, 6H). 13C NMR (75 MHz, CDCl3) δ (ppm): 170.3, 167.2, 152.6, 152.4, 157.7, 151.5, 148.2, 142.7, 135.9, 132.7, 127.1, 124.0, 123.5, 122.0, 119.5, 118.0, 117.7, 117.5, 116.6, 116.4, 111.4, 109.9, 59.5, 56.8, 40.5, 37.8, 30.9, 29.7, 26.7, 19.6. HRMS m/z calculated for C26H28F2N6O3 [M+H]+ 511.2269, found 511.2260.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-N-isopentyl-benzamide (99). Compound 99 was obtained according to general procedure C from azide 131 and 3,4-difluoro-N-isopentyl-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white solid (57 mg, 53% over 2 steps). Purity: 98%, LC tR=2.68 min, MS (ESI+): m/z=525 [M+H]+. 1H NMR (300 MHz, CDCl3) δ (ppm): 9.03 (s, 1H), 7.31-7.27 (m, 1H), 7.20-7.15 (t, J=6.96 Hz, 1H), 7.10-6.96 (m, 4H), 6.90 (s, 2H), 6.73 (s, 1H), 5.17 (s, 1H), 4.47-4.35 (m, 2H), 3.22 (s, 2H) 2.89 (s, 2H), 1.43-1.39 (m, 2H), 1.26 (s, 1H), 0.84-0.66 (m, 6H). 13C NMR (75 MHz, CDCl3) δ (ppm): 170.0, 167.5, 151.5, 149.4, 149.2, 148.3, 148.2, 142.7, 135.9, 132.4, 127.2, 124.0, 123.7, 121.8, 119.3, 117.9, 117.7, 117.5, 116.4, 111.4, 109.7, 59.5, 47.8, 40.3, 37.5, 37.0, 31.0, 25.5, 22.14. HRMS m/z calculated for C27H30F2N6O3 [M+H]+ 525.2426, found 525.2418.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-1-(1H-indol-3-ylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-N-[(1-methylimidazol-2-yl)methyl]benzamide (100). Compound 100 was obtained according to general procedure C from azide 131 and 3,4-difluoro-N-((l-methyl-1H-imidazol-2-yl)methyl)-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a brown solid (60 mg, 52% over 2 steps). Purity: 93%, LC tR=2.03 min, MS (ESI+): m/z=549 [M+H]+. 1H NMR (300 MHz, MeOD) δ (ppm): 7.47 (s, 1H), 7.36-7.25 (m, 5H), 7.04 (t, J=6.70 Hz, 1H), 6.95-6.92 (m, 3H), 6.87 (s, 1H), 5.23 (s, 1H), 4.93 (s, 1H), 4.48-4.40 (m, 3H), 3.55 (s, 2H) 3.32-3.30 (m, 3H), 2.90-2.89 (m, 2H). HRMS m/z calculated for C27H26F2N8O3 [M+H]+ 549.2174, found 549.2175.
N-[1-(1-Hydroxycarbamoylmethyl-2-phenyl-ethyl)-1H-[1,2,3]triazol-4-ylmethyl]-4-methyl-benzamide (101). Compound 101 was obtained according to general procedure B from azide 108 and 4-methyl-N-(prop-2-yn-1-yl)benzamide as a white solid (20 mg, 54%), Purity: 97%, LC tR=2.10 min, MS (ESI+): m/z=394 [M+H]+. 1H NMR (DMSO-d6) δ: 8.89 (t, J=5.6 Hz, 1H), 8.37 (s, 1H), 7.78 (s, 1H), 7.75 (d, J=8.0 Hz, 2H), 7.26 (d, J=8.0 Hz, 2H), 7.19-7.10 (m, 3H), 6.99-6.96 (m, 2H), 5.14 (m, 1H), 4.42 (d, J=5.6 Hz, 2H), 3.11 (d, J=7.6 Hz, 2H), 2.72 (dd, J=8.9 and 15.1 Hz, 1H), 2.58 (d, J=5.5 and 15.1 Hz, 1H), 2.34 (s, 3H). 13C NMR (DMSO-d6) δ (ppm): 166.4, 165.9, 145.1, 141.6, 137.4, 131.8, 129.4, 129.2 (2C), 128.7 (2C), 127.8 (2C), 127.1 (2C), 123.0, 59.3, 41.3, 37.5, 35.2, 21.4. HRMS m/z calculated for C21H24N5O3 [M+H]+ 394.1879, found 394.1892.
Methyl (3R)-4-(2-naphthyl)-3-[4-[(pyridine-4-carbonylamino)methyl]triazol-1-yl]butanoate (102). Compound 102 was obtained according to general procedure C from azide 107 and N-(prop-2-yn-1-yl)isonicotinamide followed by general procedure E as a white foam (105 mg, 98%). Purity: 100%, LC tR=2.02 min, MS (ESI+): m/z=431 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.64 (s, 1H), 7.71-7.61 (m, 5H), 7.54 (s, 1H), 7.42 (s, 1H), 7.38-7.15 (m, 2H), 7.12 (dd, J=1.2 Hz, J=8.4 Hz, 1H), 5.30-5.23 (m, 1H), 4.50 (s, 2H), 3.41-3.32 (m, 2H), 2.99-2.82 (m, 2H). 13C NMR (MeOD) δ(ppm): 168.1, 167.0, 150.6 (2C), 144.9, 143.0, 135.0, 134.4, 133.5, 129.0, 128.6 (2C), 128.3 (2C), 127.7, 126.9, 126.6, 124.8, 122.7, 61.2, 42.3, 38.8, 35.8. HRMS m/z calculated for C23H23N6O3 [M+H]+ 433.1988, found 433.2008.
N-[[1-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]-4-(hydroxycarbamoyl)benzamide (103). Compound 103 was obtained according to general procedure C from azide 107 and (E)-4-((methylimino)methyl)-N-(prop-2-yn-1-yl)benzamide followed by general procedure F as a white foam (20 mg, 67%), Purity: 100%, LC tR=2.18 min, MS (ESI+): m/z=473 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.51 (s, 1H), 7.80 (s, 4H), 7.77-7.61 (m, 4H), 7.42 (s, 1H), 7.38-7.14 (m, 2H), 7.11 (dd, J=1.5 Hz, J=8.4 Hz, 1H), 5.31-5.26 (m, 1H), 4.50 (s, 2H), 3.41-3.37 (m, 2H), 3.00-2.85 (m, 2H). 13C NMR (MeOD) δ (ppm): 168.9, 168.6, 167.1, 145.6, 138.1, 136.4, 135.4, 134.8, 133.8, 129.2, 128.8, 128.7, 128.5 (2C), 128.3 (2C), 128.0 (2C), 127.1, 126.7, 124.9, 61.5, 42.5, 38.9, 36.0. HRMS m/z calculated for C25H25N6O4 [M+H]+ 473.1985, found 473.1960.
3,4-difluoro-N-[[1-[(1R)-3-(hydroxyamino)-3-oxo-1-[[3-(trifluoromethyl)phenyl]methyl]propyl]triazol-4-yl]methyl]benzamide (104). Compound 104 was obtained according to general procedure C from azide 126 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure F as a white powder (44 mg, 62% over 2 steps), Purity 99%, LC-MS tR=2.45 min, MS (ESI+): m/z=484 [M+H]+. 1H NMR (DMSO-d6), δ (ppm): 2.62 (dd, J=5.3 Hz, J=15.2 Hz, 1H), 2.77 (dd, J=9.1 Hz, J=15.2 Hz, 1H), 3.24 (d, J=4.31 Hz, 2H), 4.21 (d, J=5.78 Hz, 2H), 5.20 (m, 1H), 7.26 (d, J=7.67 Hz, 1H), 7.31 (s, 1H), 7.39 (t, J=7.6 Hz, 1H), 7.48 (d, J=7.8 Hz, 1H), 7.56 (Dd, J=2.1 Hz, J=9.6 Hz, 1H), 7.72-7.77 (m, 1H), 7.84 (s, 1H), 7.89 (td, J=2.1 Hz, J=7.9 Hz, 1H), 8.79 (s, 1H, NH hydroxamic acid), 9.09 (t, J=5.78 Hz, 1H), 10.51 (s, 1H). 19F NMR (DMSO-d6), δ (ppm): −61.6, −135.1, −138.6. 13C NMR (DMSO-d6), δ(ppm): 34.7, 36.9, 40.0, 58.5, 116.5, 116.7, 122.7, 123.3, 124.8, 125.4, 129.2, 131.5, 131.6, 133.0, 138.3, 144.2, 163.8, 165.3. HRMS m/z calculated for C21H19F5N5O3 [M+H]+ 484.1406, found 484.1408.
N-({1-[(2R)-1-[4-(2-tert-butyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]-3-(hydroxycarbamoyl)propan2-yl]-1H-1,2,3-triazol-4-yl}methyl)-3,4-difluorobenzamide (105). Compound 105 was obtained according to general procedure C from azide 127 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure F as a white solid (61 mg, 47%). Purity 99%, LC tR=2.48 min, MS (ESI−): m/z=538 [M−H]−. 1H NMR (DMSO-d6) δ (ppm): 10.52 (s, 1H), 9.09 (t, J=5.7 Hz, 1H), 8.81 (s, 1H), 7.87 (dd, J=2.0 Hz, J=7.8 Hz, 1H), 7.85 (s, 1H), 7.82 (d, J=8.2 Hz, 2H), 7.72 (m, 1H), 7.51 (dd, J=2.0 Hz, J=8.3 Hz, 1H), 7.12 (d, J=8.2 Hz, 2H), 5.19 (m, 1H), 4.42 (d, J=5.7 Hz, 2H), 3.20 (m, 2H), 2.77 (dd, J=8.7 Hz, J=15.1 Hz, 1H), 2.66 (dd, J=5.7 Hz, J=15.1 Hz, 1H), 1.71 (s, 9H). 13C NMR (DMSO-d6) δ (ppm): 165.4, 163.8, 163.4, 144.2, 139.3, 131.5, 129.7, 126.2, 125.5, 124.7, 122.6, 117.5, 116.5, 63.8, 58.6, 40.6, 37.0, 34.8, 28.7. 19F NMR (DMSO-d6) δ (ppm): −135.1 (s), −138.4 (s). HRMS m/z calculated for C25H28F2N9O3 [M+H]+: 540.2283, found: 540.2292.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(quinolin-3-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (143). Compound 143 was obtained according to general procedure C from azide 140 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white solid (59 mg, 49%). Purity 96%, LC tR=2.03 min, MS (ESI+): m/z=467 [M+H]+. 1H NMR (MeOD) δ (ppm): 8.46 (d, J=2.0 Hz, 1H), 7.89 (d, J=2.0 Hz, 1H), 7.86 (d, J=9.1 Hz, 1H), 7.72 (s, 1H), 7.71 (dd, J=8.4 and 1.0 Hz, 1H), 7.65 (dd, J=7.0 and 1.7 Hz, 1H), 7.63-7.59 (m, 1H), 7.59-7.53 (m, 1H), 7.45 (ddd, J=8.0, 7.0 and 1.0 Hz, 1H), 7.31 (ddd, J=10.3, 8.4 and 8.2 Hz, 1H), 5.40-5.30 (m, 1H), 4.47 (d, J=4.3 Hz, 2H), 3.51 (dd, J=14.5 and 5.5 Hz, 1H), 3.46 (dd, J=14.1 and 9.2 Hz, 1H), 3.00 (dd, J=15.0 and 8.8 Hz, 1H), 2.92 (dd, J=15.0 and 5.9 Hz, 1H). 13C NMR (MeOD) δ (ppm): 168.3, 167.3, 153.7 (dd, J=252.3 and 12.6 Hz), 152.2, 151.2 (dd, J=247.5 and 13.2 Hz), 147.5, 145.8, 137.9, 132.5 (dd, J=4.8 and 3.6 Hz), 131.3, 130.8, 129.3, 128.9, 128.7, 128.2, 125.5 (dd, J=7.2 and 3.6 Hz), 125.0, 118.4 (d, J=18.0 Hz), 117.9 (d, J=18.6 Hz), 61.0, 39.3, 38.9, 36.0. HRMS m/z calculated for C23H20F2N6O3 [M+H]+ 467.1643, found 467.1642.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (144). Compound 144 was obtained according to general procedure C from azide 141 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white solid (51 mg, 67%). Purity: 96%, LC tR=2.48 min, MS (ESI+): m/z=470 [M+H]+. 1H NMR (MeOD) δ (ppm): 7.76 (ddd, J=11.2, 7.7 and 2.2 Hz, 1H), 7.71-7.65 (m, 1H), 7.62 (s, 1H), 7.36 (ddd, J=10.2, 8.4 and 8.2 Hz, 1H), 6.79 (d, J=7.7 Hz, 1H), 6.64 (br d, J=8.1 and 1.6 Hz, 1H), 6.62 (br s, 1H), 5.17-5.06 (m, 1H), 4.53 (s, 2H), 3.16-3.05 (m, 2H), 2.87 (dd, J=14.9 and 9.0 Hz, 1H), 2.77 (dd, J=14.9 and 5.6 Hz, 1H), 2.62-2.50 (m, 4H), 1.68-1.64 (m, 4H). 13C NMR (MeOD) δ (ppm): 168.6, 167.3, 153.8 (dd, J=252.3 and 12.6 Hz), 151.3 (dd, J=248.1 and 13.2 Hz), 145.4, 138.2, 136.7, 134.8, 132.7 (dd, J=4.8 and 3.6 Hz), 130.6, 130.2, 127.2, 125.6 (dd, J=7.2 and 3.6 Hz), 124.9, 118.5 (d, J=18.0 Hz), 117.9 (d, J=19.2 Hz), 61.7, 42.0, 38.8, 36.1, 30.2, 29.9, 24.30, 24.26. HRMS m/z calculated for C24H25F2N5O3 [M+H]+ 470.2004, found 470.2000.
4-fluoro-N-(1-(1-((R)-4-(hydroxyamino)-1-(naphthalen-2-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)benzamide (164). Compound 164 was obtained according to general procedure C′ from azide 109 and N-(but-3-yn-2yl)-4-fluorobenzamide, followed by general procedure G′ as a white solid (61 mg, 58% over 2 steps). Purity: 99%, LC tR=2.32 min, MS (ESI+): m/z=462 [M+H]+. Diastereoisomeric mixture, (R,S)/(R,R) ratio=4:1. 1H NMR (MeOD-d4) δ (ppm): 7.81-7.76 (m, 2H), 7.74-7.61 (m, 3H), 7.58 (s, 0.2H), 7.53 (s, 0.8H), 7.41-7.33 (m, 3H), 7.17-7.10 (m, 3H), 5.31-5.21 (m, 2H), 3.45-3.35 (m, 2H), 3.25-3.14 (m, 0.2H), 2.98 (dd, J=14.8, 9.0 Hz, 0.8H), 2.97 (dd, J=14.8, 8.9 Hz, 0.2H), 2.87 (dd, J=14.9, 5.5 Hz, 0.8H), 1.48 (d, J=7.0 Hz, 0.6H), 1.44 (d, J=7.0 Hz, 2.4H). 13C NMR (MeOD-d4) δ (ppm): major (R,S): 168.6, 168.2, 166.2 (d, J=250.2 Hz), 150.3, 135.4, 134.8, 133.9, 131.8 (d, J=2.7 Hz), 131.1 (d, J=9.0 Hz, 2C), 129.2, 128.8, 128.5 (2C), 128.0, 127.1, 126.8, 123.9, 116.3 (d, J=22.2 Hz, 2C), 61.5, 43.3, 42.5, 38.8, 20.4. Minor (R,R): 168.6, 168.2, 166.2 (d, J=250.2 Hz), 150.4, 135.4, 134.8, 133.9, 131.8 (d, J=2.7 Hz), 131.1 (d, J=9.0 Hz, 2C), 129.2, 128.8, 128.5 (2C), 128.0, 127.1, 126.8, 123.6, 116.3 (d, J=22.2 Hz, 2C), 61.5, 43.3, 42.5, 38.8, 20.5. HRMS: m/z calculated for C25H25N5O3F [M+H]+: 462.1941; found: 462.1942.
(R)-4-fluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (165). Compound 165 was obtained according to general procedure C′ from azide 141 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a white solid (35 mg, 72% over 2 steps). Purity: 99.5%, LC tR=2.40 min, MS (ESI+): m/z=452 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.92-7.85 (m, 2H), 7.62 (s, 1H), 7.23-7.14 (m, 2H), 6.79 (d, J=7.5 Hz, 1H), 6.67-6.61 (m, 2H), 5.17-5.07 (m, 1H), 4.54 (s, 2H), 3.21-3.06 (m, 2H), 2.88 (dd, J=14.9 and 9.0 Hz, 1H), 2.78 (dd, J=14.9 and 5.6 Hz, 1H), 2.64-2.48 (m, 4H), 1.73-1.58 (m, 4H). 13C NMR (MeOD-d4) δ (ppm): 168.6, 168.6, 166.3 (d, J=249.3 Hz), 145.6, 138.2, 136.7, 134.7, 131.6 (d, J=3.2 Hz), 131.0 (d, J=9.2 Hz, 2C), 130.6, 130.2, 127.1, 124.8, 116.4 (d, J=22.2 Hz, 2C), 61.6, 42.0, 38.8, 36.0, 30.2, 29.9, 24.3, 24.2. HRMS: m/z calculated for C24H27NO3F, [M+H]+: 452.2098; found: 452.2096.
(R)-4-fluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-N-methylbenzamide (166). Compound 166 was obtained according to general procedure C′ from azide 141 and N-methyl-N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as an off-white solid (28 mg, 56% over 2 steps). Purity: 99%, LC tR=2.42 min, MS (ESI+): m/z=466 [M+H]+. Mixture of amide isomer, approximate ratio: 1/0.7. 1H NMR (MeOD-d4) δ (ppm): 7.66 (br s, 0.6H), 7.47 (br s, 2.4H), 7.21-7.12 (m, 2H), 6.85 (br s, 1H), 6.70 (s, 0.8H), 6.67 (s, 1.2H), 5.15 (br s, 1H), 4.76-4.61 (m, 1H), 4.41 (br s, 1H), 3.19 (dd, J=13.8 and 5.4 Hz, 1H), 3.11 (dd, J=13.8 and 9.8 Hz, 1H), 2.97-2.79 (m, 5H), 2.68-2.54 (m, 4H), 1.70 (br s, 4H). 13C NMR (MeOD-d4) δ (ppm): major isomer: 172.6, 168.5, 164.9 (d, J=249.5 Hz), 143.6 (br), 138.2, 136.8, 134.9, 133.2 (m), 130.7 (d, J=9.4 Hz, 2C), 130.6, 130.2, 127.1, 125.5, 116.5 (d, J=22.4 Hz, 2C), 61.7, 47.5, 42.1, 38.8, 37.8, 30.2, 29.9, 24.3, 24.3. Minor isomer: 172.6, 168.5, 164.9 (d, J=249.5 Hz), 143.6 (br), 138.2, 136.8, 134.9, 133.2 (m), 130.7 (d, J=9.4 Hz, 2C), 130.6, 130.2, 127.1, 125.5, 116.5 (d, J=22.4 Hz, 2C), 61.0, 47.5, 43.3, 42.3, 33.5, 30.8, 30.2, 24.3, 24.3. HRMS: m/z calculated for C25H29N5O3F [M+H]+: 466.2254; found: 466.2248.
4-fluoro-N-(1-(1-((R)-4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)benzamide (167). Compound 167 was obtained according to general procedure C′ from azide 141 and N-(but-3-yn-2yl)-4-fluorobenzamide, followed by general procedure G′ as a white solid (50 mg, 72% over 2 steps). Purity: 95%, LC tR=2.47 min, MS (ESI+): m/z=466 [M+H]+. R,R isomer as major product, but signal of R,S isomer also visible, approximate ratio: 1:0.4. 1H NMR (MeOD-d4) δ (ppm): 7.91-7.85 (m, 2H), 7.57 (s, 1H), 7.21-7.13 (m, 2H), 6.84-6.79 (m, 1H), 6.67-6.61 (m, 2H), 5.33 (q, J=6.9 Hz, 1H), 5.14-5.05 (m, 1H), 3.20-3.05 (m, 2H), 2.89 (dd, J=14.9 and 9.1 Hz, 1H), 2.78 (dd, J=14.9 and 5.4 Hz, 1H), 2.67-2.51 (m, 4H), 1.73-1.64 (m, 4H), 1.54 (d, J=7.0 Hz, 3H). 13C NMR (MeOD-d4) δ (ppm): 168.6, 168.2, 166.2 (d, J=249.6 Hz), 150.1, 138.2, 136.8, 134.8, 131.9-131.8 (m), 131.1 (d, J=9.0 Hz, 2C), 130.6, 130.2, 127.1, 123.8, 116.3 (d, J=22.3 Hz, 2C), 61.7, 43.3, 42.1, 38.7, 37.1, 30.2, 29.9, 24.3, 24.3, 20.6. HRMS: m/z calculated for C25H29N5O3F [M+H]+: 466.2254; found: 466.2256.
3,4-difluoro-N-(1-(1-((R)-4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)benzamide (168). Compound 168 was obtained according to general procedure C′ from azide 142 and N-(but-3-yn-2yl)-3,4-difluorobenzamide as a white solid (31 mg, 29%). Purity: 99%, LC tR=2.52 min, MS (ESI+): m/z=484 [M+H]+. Mixture of (R,S) and (R,R) isomers: 1H NMR (CDCl3+1% TMS) δ (ppm): 7.81-7.73 (m, 1H), 7.72-7.65 (m, 1H), 7.35 (dtd, J=10.4 and 8.2 and 2.8 Hz, 1H), 6.82 (br t, J=7.1 Hz, 1H), 6.68-6.63 (m, 1H), 6.61 (s, 1H), 5.31 (br q, J=7.0 Hz, 1H), 5.16-5.05 (m, 1H), 3.18-3.04 (m, 2H), 2.89 (ddd, J=14.9 and 9.0 and 2.8 Hz, 1H), 2.78 (dd, J=14.9 and 5.2 Hz, 1H), 2.65-2.54 (m, 4H), 1.72-1.64 (m, 4H), 1.55+1.54 (2d, J=7.0 Hz, 3H). 13C NMR (CDCl3+1% TMS) δ (ppm): (R,S) isomer: 168.6, 166.8, 153.7 (dd, J=252.3 and 12.6 Hz), 151.3 (dd, J=248.6 and 13.2 Hz), 150.0, 138.2, 136.8, 134.8, 133.0-132.7 (m), 130.6, 130.2, 127.2, 125.7 (dd, J=7.2 and 3.6 Hz), 123.9, 118.4 (d, J=18.0 Hz), 118.0 (d, J=18.5 Hz), 61.7, 43.4, 42.1, 38.7, 30.2, 29.9, 24.3, 24.3, 20.5. (R,R) isomer: 168.6, 166.8, 153.7 (dd, J=252.3 and 12.6 Hz), 151.3 (dd, J=248.6 and 13.2 Hz), 150.1, 138.2, 136.8, 134.8, 133.0-132.7 (m), 130.6, 130.2, 127.2, 125.7 (dd, J=7.2 and 3.6 Hz), 123.6, 118.4 (d, J=18.0 Hz), 118.0 (d, J=18.5 Hz), 61.7, 43.4, 42.1, 38.7, 30.2, 29.9, 24.3, 24.3, 20.6. HRMS: m/z calculated for C25H28N5O3F2 [M+H]+=484.2160; found: 484.2160.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-N-methylbenzamide (169). Compound 169 was obtained according to general procedure C′ from azide 141 and N-methyl-N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as an off-white solid (19 mg, 18% over 2 steps). Purity: 97%, LC tR=2.50 min, MS (ESI+): m/z=484 [M+H]+. Mixture of amide isomers, approximate ratio: 1.5/1: 1H NMR (MeOD-d4) δ (ppm): 7.72 (br s, 0.6H), 7.50 (br s, 0.4H), 7.44-7.31 (m, 2H), 7.26 (br s, 1H), 6.93-6.83 (m, 1H), 6.74-6.60 (m, 2H), 5.21-5.09 (m, 1H), 4.74-4.59 (m, 1H), 4.41 (br s, 1H), 3.24-3.07 (m, 2H), 2.96-2.79 (m, 5H), 2.70-2.52 (m, 4H), 1.70 (br s, 4H). 13C NMR (MeOD-d4) δ (ppm): major isomer: 168.6, 152.5 (dd, J=249.3 and 12.6 Hz), 151.4 (dd, J=249.3 and 13.2 Hz), 143.5, 143.0, 138.2, 136.8, 134.9, 134.2-134.0 (m), 130.6, 130.2, 127.1, 125.6, 125.3 (br), 118.8 (d, J=17.9 Hz), 117.9 (d, J=18.8 Hz), 61.8, 42.1, 38.8, 37.7, 30.2, 29.9, 24.3, 24.3. Minor isomer: 168.6, 152.5 (dd, J=249.3 and 12.6 Hz), 151.4 (dd, J=249.3 and 13.2 Hz), 143.5, 143.0, 138.2, 136.8, 134.9, 134.2-134.0 (m), 130.6, 130.2, 127.1, 125.6, 125.3 (br), 118.8 (d, J=17.9 Hz), 117.9 (d, J=18.8 Hz), 61.8, 47.4, 43.3, 33.4, 30.2, 29.9, 24.3, 24.3. HRMS: m/z calculated for C25H28N5O3F2 [M+H]+: 484.2160; found: 484.2150.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(quinolin-7-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (170). Compound 170 was obtained according to general procedure C′ from azide 151 and N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as an off-white solid (24 mg, 59% over 2 steps). Purity: 98%, LC tR=1.97 min, MS (ESI+): m/z=467 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 8.70 (dd, J=4.4 and 1.6 Hz, 1H), 8.19 (dd, J=8.2 and 1.0 Hz, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.68-7.55 (m, 4H), 7.41 (dd, J=8.2 and 4.4 Hz, 1H), 7.37-7.28 (m, 2H), 5.39-5.29 (m, 1H), 4.50 (d, J=15.3 Hz, 1H), 4.44 (d, J=15.3 Hz, 1H), 3.51 (dd, J=13.8 and 5.5 Hz, 1H), 3.45 (dd, J=13.8 and 8.9 Hz, 1H), 3.00 (dd, J=15.0 and 9.0 Hz, 1H), 2.90 (dd, J=15.0 and 5.6 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.4, 167.3, 153.7 (dd, J=252.2 and 12.6 Hz), 151.3, 151.3 (dd, J=248.0 and 13.1 Hz), 148.5, 145.6, 140.4, 138.0, 132.6 (dd, J=4.1 and 3.6 Hz), 129.5, 129.3, 129.0, 128.7, 125.5 (dd, J=7.2 and 3.6 Hz), 124.9, 122.3, 118.5 (d, J=18.0 Hz), 117.9 (d, J=18.6 Hz), 61.2, 42.4, 38.9, 36.0. HRMS: m/z calculated for C23H21F2N6O3 [M+H]+: 467.1643; found: 467.1645.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-4-oxo-1-(2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (171). Compound 171 was obtained according to general procedure C′ from azide 153 and N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as an off-white solid (29 mg, 58% over 2 steps). Purity: 97%, LC tR=1.78 min, MS (ESI+): m/z=485 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.75 (ddd, J=11.2 and 7.7 and 2.1 Hz, 1H), 7.69-7.62 (m, 1H), 7.64 (s, 1H), 7.36 (ddd, J=10.2 and 8.4 and 8.2 Hz, 1H), 6.82-6.77 (m, 1H), 6.78 (s, 1H), 6.68-6.64 (m, 1H), 5.19-5.09 (m, 1H), 4.52 (s, 2H), 3.19 (dd, J=13.8 and 5.7 Hz, 1H), 3.13 (dd, J=13.8 and 9.2 Hz, 1H), 2.90 (dd, J=15.1 and 8.8 Hz, 1H), 2.80 (dd, J=15.0 and 5.7 Hz, 1H), 2.79-2.68 (m, 2H), 2.43 (t, J=7.5 Hz, 2H). 13C NMR (MeOD-d4) δ (ppm): 173.7, 168.5, 167.4, 153.8 (dd, J=252.3 and 12.6 Hz), 151.4 (dd, J=248.1 and 13.2 Hz), 145.5, 137.7, 132.7-132.5 (m), 132.6, 129.5, 129.0, 125.5 (dd, J=7.3 and 3.6 Hz), 125.3, 124.9, 118.6 (d, J=18.0 Hz), 117.9 (d, J=18.6 Hz), 116.6, 61.6, 41.7, 38.7, 36.1, 31.4, 26.0. HRMS: m/z calculated for C23H23N6O4F2 [M+H]+: 485.1749; found: 485.1742.
4-fluoro-N-(1-(1-((R)-4-(hydroxyamino)-4-oxo-1-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)butan-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)benzamide (172). Compound 172 was obtained according to general procedure C′ from azide 154 and N-(but-3-yn-2yl)-4-fluorobenzamide, followed by general procedure G′ as a white solid (75 mg, 59% over 2 steps). Purity: 99%, LC tR=2.82 min, MS (ESI+): m/z=522 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.90-7.85 (m, 2H), 7.60 (s, 1H), 7.19-7.11 (m, 3H), 6.83 (br s, 1H), 6.79 (dd, J=7.9 and 1.4 Hz, 1H), 5.33 (q, J=6.9 Hz, 1H), 5.16-5.06 (m, 1H), 3.26-3.10 (m, 2H), 2.89 (dd, J=14.9 and 9.0 Hz, 1H), 2.79 (dd, J=14.9 and 5.5 Hz, 1H), 1.61 (br s, 4H), 1.56+1.52 (2d, J=6.9 Hz, 3H), 1.18+1.09 (2s, 12H). 13C NMR (MeOD-d4) δ (ppm): (R,R) isomer: 168.6, 168.2, 166.2 (d, J=249.1 Hz), 150.0, 146.0, 144.5, 134.8, 131.9-131.8 (m), 131.1 (d, J=9.0 Hz, 2C), 128.2, 127.8, 127.3, 123.9, 116.3 (d, J=22.2 Hz, 2C), 61.7, 43.3, 42.0, 38.9, 36.1 (2C), 35.0, 34.8, 32.2 (4C), 20.6. (R,S) isomer: 168.6, 168.2, 166.2 (d, J=249.1 Hz), 150.1, 146.0, 144.5, 135.0, 131.9-131.8 (m), 131.1 (d, J=9.0 Hz, 2C), 128.2, 127.8, 127.3, 123.7, 116.3 (d, J=22.2 Hz, 2C), 60.8, 43.3, 42.3, 38.9, 36.1 (2C), 35.0, 34.8, 32.3 (4C), 20.7. HRMS: m/z calculated for C29H37O3N5F [M+H]+: 522.2880; found: 522.2888.
(R)—N-((1-(1-(benzo[b]thiophen-5-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-4-fluorobenzamide (173). Compound 173 was obtained according to general procedure C′ from azide 155 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a white solid (47 mg, 70% over 2 steps). Purity: 99.5%, LC tR=2.20 min, MS (ESI+): m/z=454 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.85-7.78 (m, 2H), 7.66 (d, J=8.2 Hz, 1H), 7.61 (s, 1H), 7.46-7.44 (m, 2H), 7.21-7.13 (m, 3H), 6.96 (dd, J=8.2 and 1.1 Hz, 1H), 5.29-5.19 (m, 1H), 4.54 (dd, J=15.5 and 4.1 Hz, 1H), 4.47 (dd, J=15.5 and 4.1 Hz, 1H), 3.43-3.28 (m, 2H), 2.94 (dd, J=14.9 and 8.9 Hz, 1H), 2.84 (dd, J=14.9 and 5.6 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.7, 168.5, 166.2 (d, J=250.4 Hz), 145.7, 141.4, 139.8, 133.9, 131.6 (d, J=3.3 Hz), 131.0 (d, J=9.0 Hz, 2C), 128.0, 126.3, 124.9, 124.8, 124.6, 123.4, 116.4 (d, J=22.2 Hz, 2C), 61.8, 42.2, 38.8, 36.0. HRMS: m/z calculated for C22H22N5O3SF [M+H]+: 454.1349; found 454.1348.
(R)—N-((1-(1-(benzo[b]thiophen-6-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-4-fluorobenzamide (174). Compound 174 was obtained according to general procedure C′ from azide 156 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a white solid (43 mg, 73% over 2 steps). Purity: 99.5%, LC tR=2.20 min, MS (ESI+): m/z=454 [M+H]+. 1H NMR, MeOD, δ (ppm): 7.84-7.78 (m, 2H), 7.66-759 (m, 2H), 7.51 (br s, 1H), 7.43 (d, J=5.4 Hz, 1H), 7.20 (dd, J=5.4, 1.0 Hz, 1H), 7.19-7.12 (m, 2H), 6.98 (dd, J=8.2, 1.3 Hz, 1H), 5.29-5.18 (m, 1H), 4.52 (d, J=15.8 Hz, 1H), 4.47 (d, J=15.8 Hz, 1H), 3.38 (dd, J=13.8, 5.6 Hz, 1H), 3.32 (dd, J=13.8, 9.0 Hz, 1H), 2.93 (dd, J=14.9, 9.0 Hz, 1H), 2.83 (dd, J=14.9, 5.5 Hz, 1H). 13C NMR, MeOD, δ(ppm): 168.8, 168.5, 166.2 (d, J=250.6 Hz), 145.7, 141.4, 140.0, 134.1, 131.6 (d, J=2.8 Hz), 131.0 (d, J=9.0 Hz, 2C), 127.4, 126.4, 124.9, 124.6, 124.5, 123.6, 116.3 (d, J=22.2 Hz, 2C), 61.7, 42.3, 38.8, 36.0. HRMS: m/z calculated for C22H21N5O3SF, [M+H]+: 454.1349; found: 454.1351.
(R)-3,4-difluoro-N-((1-(4-(hydroxyamino)-1-(1H-indol-5-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (175). Compound 175 was obtained according to general procedure C′ from azide 157 and N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as an off-white solid (37 mg, 54% over 2 steps). Purity: 97%, LC tR=2.07 min, MS (ESI+): m/z=455 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.68 (ddd, J=11.3, 7.7, 2.1 Hz, 1H), 7.62-7.56 (m, 1H), 7.55 (s, 1H), 7.34 (ddd, J=10.2, 8.5, 8.0 Hz, 1H), 7.19 (br d, J=8.3 Hz, 1H), 7.15 (d, J=0.7 Hz, 1H), 7.12 (d, J=3.1 Hz, 1H), 6.76 (dd, J=8.3, 1.6 Hz, 1H), 6.25 (dd, J=3.1, 0.7 Hz, 1H), 5.23-5.14 (m, 1H), 4.53 (dd, J=16.1, 4.3 Hz, 1H), 4.46 (dd, J=16.1, 4.3 Hz, 1H), 3.30 (dd, J=13.5, 5.8 Hz, 1H), 3.24 (dd, J=13.5, 8.6 Hz, 1H), 2.91 (dd, J=14.9, 9.1 Hz, 1H), 2.81 (dd, J=14.9, 5.5 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.8, 167.5, 153.7 (dd, J=151.2 and 12.8 Hz), 151.3 (dd, J=247.9 and 13.1 Hz), 145.2, 136.7, 132.7 (dd, J=4.8 and 3.2 Hz), 129.6, 128.0, 126.0, 125.5 (dd, J=7.2 and 4.2 Hz), 124.9, 123.2, 121.5, 118.5 (d, J=17.9 Hz), 117.9 (d, J=18.4 Hz), 112.2, 102.1, 62.3, 42.8, 38.7, 36.1. HRMS: m/z calculated for C22H21N6O3F2, [M+H]+: 455.1643; found: 455.1641.
(R)-4-fluoro-N-((1-(4-(hydroxyamino)-1-(1H-indol-6-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (176). Compound 176 was obtained according to general procedure C′ from azide 158 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a white solid (41 mg, 73% over 2 steps). Purity: 99.5%, LC tR=2.08 min, MS (ESI+): m/z=437 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.83-7.77 (m, 2H), 7.54 (s, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.17-7.13 (m, 2H), 7.11 (d, J=3.1 Hz, 1H), 7.00 (s, 1H), 6.65 (dd, J=8.0 and 1.0 Hz, 1H), 6.30 (d, J=3.1 Hz, 1H), 5.25-5.14 (m, 1H), 4.53 (d, J=15.6 Hz, 1H), 4.47 (d, J=15.6 Hz, 1H), 3.33-3.22 (m, 2H), 2.91 (dd, J=14.9 and 8.9 Hz, 1H), 2.80 (dd, J=14.9 and 5.4 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.9, 168.7, 166.2 (d, J=250.3 Hz), 145.4, 137.8, 131.7 (d, J=3.2 Hz), 131.0 (d, J=9.6 Hz, 2C), 130.6, 128.4, 125.7, 124.9, 121.2 (2C), 116.4 (d, J=22.2 Hz, 2C), 112.5, 102.1, 62.2, 42.8, 38.8, 36.0. HRMS: m/z calculated for C22H22N6O3F [M+H]+: 437.1735; found: 437.1737.
(R)-4-fluoro-N-((1-(4-(hydroxyamino)-1-(1-methyl-1H-indol-5-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (177). Compound 177 was obtained according to general procedure C′ from azide 159 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a light yellow wax (6 mg, 14% over 2 steps). Purity: 97%, LC tR=2.23 min, MS (ESI+): m/z=451 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.83-7.79 (m, 2H), 7.53 (s, 1H), 7.20-7.12 (m, 4H), 7.03 (d, J=3.0 Hz, 1H), 6.78 (dd, J=8.3 and 1.5 Hz, 1H), 6.23 (dd, J=3.0 and 0.4 Hz, 1H), 5.23-5.14 (m, 1H), 4.50 (s, 2H), 3.66 (s, 3H), 3.34-3.20 (m, 2H), 2.90 (dd, J=14.9 and 9.0 Hz, 1H), 2.80 (dd, J=14.9 and 5.5 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.8 (2C), 166.2 (d, J=250.8 Hz), 145.5, 137.3, 131.7 (d, J=3.6 Hz), 131.0 (d, J=9.0 Hz, 2C), 130.4, 130.2, 128.1, 124.8, 123.3, 121.9, 116.3 (d, J=22.2 Hz, 2C), 110.2, 101.4, 62.3, 42.7, 38.7, 36.0, 32.8. HRMS: m/z calculated for C23H24N6O3F [M+H]+: 451.1894; found: 451.1893.
(R)—N-((1-(1-(benzo[d]thiazol-6-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-3,4-difluorobenzamide (178). Compound 178 was obtained according to general procedure C′ from azide 160 and N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as a colourless solid (15 mg, 55% over 2 steps). Purity: 98%, LC tR=1.95 min, MS (ESI+): m/z=473 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 9.11 (s, 1H), 7.85 (d, J=8.4 Hz, 1H), 7.68 (ddd, J=11.2 and 7.7 and 2.1 Hz, 1H), 7.68 (br s, 1H), 7.63 (s, 1H), 7.63-7.57 (m, 1H), 7.35 (ddd, J=10.3 and 8.4 and 8.1 Hz, 1H), 7.20 (dd, J=8.4 and 1.5 Hz, 1H), 5.34-5.23 (m, 1H), 4.48 (s, 2H), 3.44 (dd, J=13.7 and 5.5 Hz, 1H), 3.38 (dd, J=13.7 and 9.2 Hz, 1H), 2.95 (dd, J=14.9 and 8.8 Hz, 1H), 2.86 (dd, J=14.9 and 5.6 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.4, 167.4, 156.9, 153.8 (dd, J=252.3 and 12.6 Hz), 151.3 (dd, J=248.0 and 13.2 Hz), 145.6, 136.1, 135.2, 132.6 (dd, J=4.8 and 3.6 Hz), 128.7, 125.5 (dd, J=7.2 and 3.6 Hz), 124.9, 123.8, 123.5, 118.5 (d, J=18.0 Hz), 117.9 (d, J=18.6 Hz), 61.5, 42.1, 38.8, 36.0. HRMS: m/z calculated for C21H19N6O3SF2 [M+H]+: 473.1207; found: 473.1209.
(R)—N-((1-(1-(benzo[b]thiophen-3-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-3,4-difluorobenzamide (179). Compound 179 was obtained according to general procedure C′ from azide 161 and N-(prop-2-ynyl)-3,4-difluorobenzamide followed by general procedure G′ as an off-white solid (25 mg, 45% over 2 steps). Purity: 98%, LC tR=2.30 min, MS (ESI+): m/z=472 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.82-7.72 (m, 2H), 7.70 (dd, J=7.6 and 2.2 Hz, 1H), 7.66-7.61 (m, 1H), 7.55 (s, 1H), 7.40-7.24 (m, 3H), 7.00 (s, 1H), 5.37-5.27 (m, 1H), 4.50 (d, J=15.5 Hz, 1H), 4.45 (d, J=15.5 Hz, 1H), 3.54 (dd, J=14.5 and 5.7 Hz, 1H), 3.47 (dd, J=14.5 and 9.0 Hz, 1H), 2.98 (dd, J=14.9 and, 8.2 Hz, 1H), 2.91 (dd, J=14.9 and 6.4 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.5, 167.4, 153.8 (dd, J=252.2 and 12.6 Hz), 151.3 (dd, J=248.2 and 13.2 Hz), 145.6, 141.6, 139.6, 132.8-132.5 (m), 132.2, 125.6 (dd, J=7.6 and 3.7 Hz), 125.5, 125.5, 125.2, 124.8, 123.7, 122.4, 118.5 (d, J=18.3 Hz), 118.0 (d, J=19.2 Hz), 60.0, 38.8, 36.0, 35.0. HRMS: m/z calculated for C22H20N5O3SF2, [M+H]+: 472.1255; found: 472.1254.
(R)—N-((1-(1-(benzo[b]thiophen-3-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-4-fluorobenzamide (180). Compound 180 was obtained according to general procedure C′ from azide 161 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as an off-white solid (21 mg, 41% over 2 steps). Purity: 99.5%, LC tR=2.22 min, MS (ESI+): m/z=454 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.88-7.80 (m, 2H), 7.76-7.72 (m, 2H), 7.55 (s, 1H), 7.34 (ddd, J=7.9 and 7.1 and 1.2 Hz, 1H), 7.27 (ddd, J=7.9 and 7.1 and 1.2 Hz, 1H), 7.22-7.14 (m, 2H), 7.01 (s, 1H), 5.37-5.27 (m, 1H), 4.51 (d, J=15.5 Hz, 1H), 4.46 (d, J=15.5 Hz, 1H), 3.54 (dd, J=14.6 and 5.7 Hz, 1H), 3.47 (dd, J=14.6 and 9.3 Hz, 1H), 2.97 (dd, J=14.9 and 8.1 Hz, 1H), 2.91 (dd, J=14.9 and 6.4 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.8, 168.6, 166.3 (d, J=250.3 Hz), 145.8, 141.6, 139.7, 132.2, 131.6 (d, J=3.4 Hz), 131.0 (d, J=9.0 Hz, 2C), 125.5, 125.5, 124.7, 123.8, 122.4, 116.3 (d, J=22.1 Hz, 2C), 60.0, 38.8, 36.0, 35.0. HRMS: m/z calculated for C22H21N5O3FS [M+H]+: 454.1349; found: 454.1346.
N-(1-(1-((R)-1-(benzo[b]thiophen-3-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)-4-fluorobenzamide (181). Compound 181 was obtained according to general procedure C′ from azide 161 and N-(but-3-yn-2yl)-4-fluorobenzamide, followed by general procedure G′ as a white solid (23 mg, 48% over 2 steps). Purity: 95%, LC tR=2.28 min, MS (ESI+): m/z=468 [M+H]+. Mixture of diastereoisomers, estimated (R,R)/(R,S) ratio: 2.3/1 1H NMR (MeOD-d4) δ (ppm): 7.87-7.75 (m, 3.2H), 7.71 (br d, J=7.5 Hz, 0.8H), 7.52 (s, 0.3H), 7.49 (s, 0.7H), 7.38-7.26 (m, 2H), 7.21-7.12 (m, 2H), 7.03 (s, 0.7H), 7.02 (s, 0.3H), 5.37-5.22 (m, 1H), 5.26 (q, J=7.1 Hz, 1H), 3.60-3.43 (m, 2H), 3.04-2.89 (m, 2H), 1.49 (d, J=7.1 Hz, 0.9H), 1.46 (d, J=7.1 Hz, 2.1H). 13C NMR (MeOD-d4) δ (ppm): Major isomer, (R,R): 168.6, 168.3, 166.2 (d, J=250.0 Hz), 150.3, 141.6, 139.7, 132.3, 131.9 (d, J=3.2 Hz), 131.1 (d, J=9.0 Hz, 2C), 125.5, 125.5, 125.3, 123.8, 123.7, 122.4, 116.3 (d, J=22.1 Hz, 2C), 60.1, 43.3, 38.7, 35.1, 20.4. Minor isomer, (R,S): 168.6, 168.3, 166.2 (d, J=250.0 Hz), 150.5, 141.6, 139.7, 132.2, 131.9 (d, J=3.2 Hz), 131.1 (d, J=9.0 Hz, 2C), 125.6, 125.5, 125.3, 123.8, 123.5, 122.4, 116.3 (d, J=22.1 Hz, 2C), 60.0, 43.3, 38.7, 35.1, 20.6. HRMS: m/z calculated for C23H23NO3FS [M+H]+: 468.1506; found: 468.1499.
(S)—N-((1-(1-(benzo[b]thiophen-2-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-4-fluorobenzamide (182). Compound 182 was obtained according to general procedure C′ from azide 162 and N-(prop-2-ynyl)-4-fluorobenzamide followed by general procedure G′ as a purple-brown solid (9 mg, 34% over 2 steps). Purity: 97%, LC tR=2.23 min, MS (ESI+): m/z=454 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.83-7.77 (m, 2H), 7.56 (s, 1H), 7.67-7.61 (m, 1H), 7.57-7.51 (m, 1H), 7.24-7.11 (m, 4H), 6.89 (s, 1H), 5.33-5.23 (m, 1H), 4.54 (s, 2H), 3.61 (dd, J=14.7 and 8.6 Hz, 1H), 3.55 (dd, J=14.6 and 5.6 Hz, 1H), 2.94 (dd, J=15.1 and 8.9 Hz, 1H), 2.87 (dd, J=15.1 and 5.8 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 167.4, 166.9, 164.8 (d, J=250.0 Hz), 144.5, 139.8, 139.6, 139.1, 130.2 (d, J=3.6 Hz), 129.6 (d, J=9.0 Hz, 2C), 123.9, 123.7, 123.6, 123.1, 122.7, 121.5, 114.9 (d, J=22.2 Hz, 2C), 59.7, 37.4, 35.5, 34.6. HRMS: m/z calculated for C22H21N5O3FS [M+H]+: 454.1349; found: 454.1345.
(R)—N-((1-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-4-(hydroxyamino)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-3,4-difluorobenzamide (183). Compound 183 was obtained according to general procedure C′ from azide 163 and N-(prop-2-ynyl)-3,4-difluorobenzamide as a beige solid (90 mg, 60%). Purity: 99%, LC tR=2.08 min, MS (ESI+): m/z=474 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 7.76 (ddd, J=11.2 and 7.7 and 2.1 Hz, 1H), 7.70-7.64 (m, 1H), 7.62 (s, 1H), 7.36 (ddd, J=10.2 and 8.4 and 8.2 Hz, 1H), 6.56 (d, J=8.2 Hz, 1H), 6.48 (d, J=1.8 Hz, 1H), 6.38 (dd, J=8.2 and 1.8 Hz, 1H), 5.15-5.05 (m, 1H), 4.59-4.46 (m, 2H), 4.10 (s, 4H), 3.12 (dd, J=13.9 and 5.7 Hz, 1H), 3.06 (dd, J=13.9 and 9.1 Hz, 1H), 2.87 (dd, J=14.9 and 9.1 Hz, 1H), 2.77 (dd, J=14.9 and 5.5 Hz, 1H). 13C NMR (MeOD-d4) δ (ppm): 168.6, 167.5, 153.8 (dd, J=252.9 and 13.2 Hz), 151.3 (dd, J=248.1 and 12.6 Hz), 145.4, 144.9, 144.0, 132.8-132.7 (m), 130.8, 125.6 (dd, J=7.2 and 3.6 Hz), 124.9, 122.8, 118.6, 118.5 (d, J=19.0 Hz), 118.1, 118.0 (d, J=20.0 Hz), 65.5 (2C), 61.6, 41.7, 38.7, 36.1. HRMS: m/z calculated for C22H22N5O5F2, [M+H]+: 474.1589; found: 474.1592.
(R)-3-fluoro-N-((1-(4-(hydroxyamino)-1-(naphthalen-2-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (184). Compound 184 was obtained according to general procedure C′ from azide 109 and 3-fluoro-N-prop-2-ynyl-benzamide followed by general procedure G′ as a white solid (34 mg, 27% over 2 steps). Purity: 99%, LC tr=2.37 min, MS (ESI+): m/z=448 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.5 (br s, 1H), 9.07 (t, J=5.8 Hz, 1H), 8.8 (s, 1H), 7.9 (s, 1H), 7.79-7.61 (m, 5H), 7.56-7.49 (m, 1H), 7.36 (br s, 1H), 7.44-7.35 (m, 3H), 7.13 (dd, J=8.5 and 1.5 Hz, 1H), 5.27 (quint, J=7.0 Hz, 1H), 4.43 (d, J=5.8 Hz, 2H), 3.31-3.28 (m, 2H), 2.78 (dd, J=15.0 and 9.0 Hz, 1H), 2.66 (dd, J=15.0 and 5.5 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.0, 165.1, 162.5 (d, J=250.0 Hz), 144.7, 137.0 (d, J=6.0 Hz), 135.0, 133.3, 132.3, 130.9 (d, J=8.0 Hz), 128.2, 127.8, 127.7, 126.4, 126.0, 118.6 (d, J=22.0 Hz), 114.6 (d, J=22.0 Hz), 59.2, 41.3, 37.6, 35.3. HRMS m/z calculated for C24H23N5O3F [M+H]+ 448.1785, found 448.1785.
(R)-4-fluoro-N-((1-(4-(hydroxyamino)-1-(naphthalen-2-yl)-4-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-N-methylbenzamide (185). Compound 185 was obtained according to general procedure C′ from azide 109 and 4-fluoro-N-methyl-N-prop-2-ynyl-benzamide followed by general procedure G′ as a white solid (58 mg, 45% over 2 steps). Purity: 99%, LC tr=2.38 min, MS (ESI+): m/z=462 [M+H]+. 1H NMR (MeOD-d4) δ(ppm): 7.73-7.61 (m, 3H), 7.56 (br s, 1H), 7.41-7.30 (m, 4H), 5.21 (dd, J=8.4 and 1.5 Hz, 2H), 7.13-6.96 (m, 2H), 5.32 (br s, 1H), 4.68 (d, J=15.4 Hz, 0.6H), 4.47 (d, J=15.4 Hz, 0.7H), 4.32 (br s, 0.7H), 3.47 (dd, J=14.0 and 5.0 Hz, 1H), 3.38-3.35 (m, 1H), 3.05 (dd, J=15.0 and 8.0 Hz, 1H), 2.94 (dd, J=15.0 and 5.0 Hz, 1H), 2.73 (br s, 1H) 2.55 (br s, 2H). 13C NMR (MeOD-d4) δ(ppm): 171.1, 167.1, 163.7 (d, J=248.9 Hz), 142.2, 142.0, 134.1, 133.4, 132.4, 131.6, 129.2, 127.9, 127.4, 127.2, 127.1, 126.6, 125.8, 125.6, 124.4, 115.1 (d, J=22.2 Hz, 2C), 60.2, 41.8, 41.2, 37.5, 36.0. HRMS m/z calculated for C25H25N5O3F [M+H]+ 462.1941, found 462.1944.
(3R)-3-(4-(3-(4-fluorobenzoyl)thiazolidin-4-yl)-1H-1,2,3-triazol-1-yl)-N-hydroxy-4-(naphthalen-2-yl)butanamide (186). Compound 186 was obtained according to general procedure C′ from azide 109 and (4-ethynylthiazolidin-3-yl)-(4-fluorophenyl)methanone followed by general procedure G′ as a white solid (82 mg, 59% over 2 steps). Purity: 95%, LC tr=2.57 min, MS (ESI+): m/z=506 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.56 (s, 1H), 8.85 (s, 1H), 8.05 (m, 1H), 7.76 (m, 3H), 7.45 (m, 6H), 7.15 (m, 3H), 5.26 (m, 1H), 4.28 (m, 1H), 3.53 (m, 5H), 3.09 (m, 1H), 2.82 (m, 2H). 13C NMR (DMSO-d6) δ(ppm): 172.6 (min), 171.1 (maj), 168.1, 166.0, 163.5 (d, J=250.0 Hz), 143.3, 146.2 (d, J=16.0 Hz), 135.0, 134.9, 133.3, 132.7, 132.3, 130.2, 128.2, 127.9, 127.8, 127.7, 126.5, 126.1, 115.9, 115.6, 59.6, 55.4, 41.6, 37.6, 35.8. HRMS m/z calculated for C26H25N5O3FS [M+H]+ 506.1662, found 506.1654.
(3R)-3-(4-(1-(4-fluorobenzoyl)piperidin-2-yl)-1H-1,2,3-triazol-1-yl)-N-hydroxy-4-(naphthalen-2-yl)butanamide (187). Compound 187 was obtained according to general procedure C′ from azide 109 and (2-ethynyl-1-piperidyl)-(4-fluorophenyl)methanone followed by general procedure G′ as a white solid (105 mg, 73% over 2 steps). Purity: 95%, LC tr=2.58 min, MS (ESI+): m/z=502 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.58 (s, 1H), 8.85 (s, 1H), 7.85-7.65 (m, 4H), 7.50-7.30 (m, 5H), 7.28-7.09 (m, 3H), 5.35-5.21 (m, 1H), 4.80-3.80 (m, 1H), 3.45-3.35 (m, 2H), 3.30-3.14 (m, 1H), 2.99-2.78 (m, 2H) 2.32-1.05 (m 7H). 13C NMR (DMSO-d6) δ (ppm): 169.1, 169.0, 166.0, 162.9 (d, J=249.5 Hz) 144.9, 135.0, 133.3, 132.3, 129.4, 128.2, 128.1, 127.8, 127.7, 127.6, 126.4, 126.0, 123.7, 115.8 (d, J=21.5 Hz), 59.5, 45.6, 41.7, 37.7, 28.1, 25.5, 19.6, 19.3. HRMS m/z calculated for C28H28N5O3F [M+H]+ 502.2254, found 502.2246.
Ethyl 5-[[(4-fluorobenzoyl)amino]methyl]-3-[(1R)-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazole-4-carboxylate (45). Azide 110 (226 mg, 0.89 mmol, 1 eq.) and ethyl 4-(N-(tert-butoxycarbonyl)-4-fluorobenzamido)but-2-ynoate (373 mg, 1.07 mmol, 1.2 eq.) were added in dioxane (2.5 mL) in a microwave vessel and the resulting mixture was heated 6 h at 130° C. (200 W). After cooling, the solvent was evaporated and the residue was treated 2 h at room temperature with a solution of TFA in dichloromethane (10%, 20 mL). TFA and dichloromethane were evaporated and the crude was purified by flash chromatography on silica gel (CH2Cl2/MeOH 100:0 to 95:5 (v/v)) to afford a mixture of two regioisomers (215 mg). Then, to a solution of this mixture in DMF (5 mL) were added HOBt (300 mg, 1.95 mmol, 2.3 eq.), EDCI (238 mg, 1.24 mmol, 1.5 eq.) and NMM (0.514 mL, 4.7 mmol, 5.6 eq.). After 15 min of stirring, NH2—OTBDMS (173 mg, 1.17 mmol, 1.4 eq.) was finally introduced and the reaction mixture was stirred overnight. The solvent was evaporated and the crude was purified by flash chromatography on silica gel (CH2Cl2/MeOH 100:0 to 95:5 (v/v)) to afford compound 45 as a white solid (33 mg, 7%). Purity: 95%, LC tR=2.95 min, MS (ESI+) m/z=520 [M+H]+. 1H NMR (DMSO) δ (ppm): 1.27 (t, J=7.0 Hz, 3H), 2.73 (dd, J=4.5 and 15.6 Hz, 1H), 2.93 (dd, J=10.2 and 15.0 Hz, 1H), 4.28 (q, J=7.0 Hz, 2H), 4.35-4.40 (m, 1H), 4.95 (dd, J=6.4 and 14.7, 1H), 5.40-5.42 (m, 1H), 7.20-7.27 (m, 3H), 7.42-7.45 (m, 2H), 7.53 (br s, 1H), 7.64-7.69 (m, 2H), 7.76-7.79 (m, 1H), 7.86-7.91 (m, 2H), 8.77 (br s, 1H), 8.81 (br s, 1H), 10.62 (br s, 1H). 13C NMR (DMSO) δ (ppm): 14.5, 29.5, 31.4, 41.2, 57.1, 61.0, 115.6 (d, JC-F=21 Hz), 126.2, 126.6, 127.6, 127.8, 127.9, 128.2, 128.4, 130.5 (d, JC-F=9 Hz), 132.3, 133.3, 134.5, 136.6, 139.0, 161.2, 164.4 (d, JC-F=247 Hz), 165.4, 165.9. HRMS m/z calculated for C27H27FN5O5 [M+H]+ 520.1996, found 520.1977.
Methyl 3-(4-((3,4-difluorobenzamido)methyl)-1H-1,2,3-triazol-1-yl)acrylate (134). Compound 134 was obtained according to general procedure C from azide 133 as a yellow solid. LC tR: 2.39 min, MS (ESI+) m/z=323 [M+H]+.
3,4-difluoro-N-((1-(3-(hydroxyamino)-1-(naphthalen-2-ylthio)-3-oxopropyl)-1H-1,2,3-triazol-4-yl)methyl)benzamide (59). Compound 159 was obtained according to general procedure R followed by general procedure F as a white solid (15 mg, 25%). Purity: 99%, LC tR: 2.87 min, MS (ESI+) m/z=484 [M+H]+. HRMS m/z calculated for C23H19F2N5O3S [M+H]+ 484.1255, found 484.1255.
N-[[1-[1-(4-chlorophenyl)sulfanyl-3-(hydroxyamino)-3-oxo-propyl]triazol-4-yl]methyl]-3,4-difluoro-benzamide (67). Compound 67 was obtained according to general procedure R followed by general procedure F as a white foam (19 mg, 44%), Purity: 100%, LC tR=2.48 min, MS (ESI+): m/z=468 [M+H]+. 1H NMR (MeOD) δ (ppm): 3.05 (dd, J=6.7 Hz, J=15.1 Hz, 1H), 3.15 (dd, J=8.2 Hz, J=15.1 Hz, 1H), 4.55 (s, 2H), 6.25 (dd, J=6.7 Hz, J=6.8 Hz, 1H), 7.23 (s, 4H), 7.37 (td, J=8.2 Hz, J=10.3 Hz, 1H), 7.70 (dddd, J=2.0 Hz, J=4.2 Hz, J=5.6 Hz, J=8.0 Hz, 1H), 7.78 (ddd, J=2.1 Hz, J=7.6 Hz, J=11.3 Hz, 1H), 7.85 (s, 1H). 13C NMR (MeOD) δ (ppm): 36.2, 37.7, 65.3, 117.9 (d, J=18.8 Hz), 118.6 (d, J=18.2 Hz), 124.0, 125.6 (dd, J=3.7 Hz, J=7.3 Hz), 130.2, 130.5 (2C), 132.6, 136.9, 137.2 (2C), 146.4, 151.4 (dd, J=13.0 Hz, J=247.8 Hz), 153.8 (dd, J=13.0 Hz, J=252.5 Hz), 166.8, 167.5. 19F NMR (MeOD) δ (ppm): −136.1 (d, J=20.5), −139.8 (d, J=20.5). HRMS m/z calculated for C19H17C1F2SN5O3 [M+H]+ 468.10709, found 468.0758.
3,4-difluoro-N-[[1-[(1R,2S)-2-hydroxy-3-(hydroxyamino)-1-(2-naphthylmethyl)-3-oxo-propyl]triazol-4-yl]methyl]benzamide (84). Compound 84 was obtained according to general procedure C from azide 119 and 3,4-difluoro-N-(prop-2-yn-1-yl)benzamide followed by general procedure G as a white foam (8 mg, 10% over 2 steps). Purity: 100%, LC tR=2.40 min, MS (ESI+): m/z=482 [M+H]. 1H NMR (MeOD) δ (ppm): 3.47 (d, J=7.8 Hz, 1H), 4.50 (br s, 1H), 4.52 (q, J=15.0 Hz, 2H), 5.36 (ddd, J=3.6 Hz, J=7.7 Hz, J=8.1 Hz, 1H), 7.26 (m, 4H), 7.51 (br s, 1H), 7.60-7.76 (m, 5H), 7.98 (s, 1H). 13C NMR (DMSO-d6) δ (ppm): 34.8, 37.2, 64.2, 71.3, 116.7 (d, J=18.4 Hz), 117.5 (d, J=17.2 Hz), 123.1, 124.8 (dd, J=3.3 Hz, J=7.4 Hz), 125.5, 125.9, 127.1, 127.2, 127.3, 127.4, 127.7, 131.6, 131.8, 132.8, 134.6, 143.9, 149.1 (dd, J=12.9 Hz, J=246.5 Hz), 153.3 (dd, J=10.3 Hz, J=250.8 Hz), 163.8, 166.9. 19F NMR (MeOD) δ (ppm): −139.9 (d, J=20.3 Hz), −136.3 (d, J=20.3 Hz). HRMS m/z calculated for C24H22F2N5O4 [M+H]+ 482.1640, found 482.1622.
(R)-3-(5-Aminomethyl-[1,2,3]triazol-1-yl)-4-naphthalen-2-yl-butyric acid (114). (R)-3-Azido-4-naphthalen-2-yl-butyric acid (110) (1.85 g, 7.25 mmol) and propargylamine (0.56 mL, 8.70 mmol) were dissolved in EtOAc (55 mL), and the resulting solution was stirred at room temperature for 10 min. DMTMM (2.41 g, 8.70 mmol) was added and the reaction was stirred at room temperature for 3 h. The mixture was washed twice with 1M HCl (aq), twice with saturated NaHCO3 (aq) and twice with saturated NaCl (aq), dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using CH2Cl2 as eluent. (R)-3-Azido-4-naphthalen-2-yl-N-prop-2-ynyl-butyramide 112 was obtained as a yellowish oil (1.270 g, 60%). Purity: 90%, LC tR=2.85 min (method A), MS (ESI+): m/z=293 [M+H]+. 1H NMR (CDCl3) δ (ppm): 7.85-7.80 (m, 3H), 7.69 (s, 1H), 7.52-7.45 (m, 2H), 7.37 (dd, J=1.6 and 8.4 Hz, 1H), 5.80 (sl, 1H), 4.29-4.23 (m, 1H), 4.15-3.99 (m, 2H), 3.06 (d, J=6.8 Hz, 1H), 2.45 (dd, J=4.5 and 14.9 Hz, 1H), 2.31 (dd, J=8.6 and 14.9 Hz, 1H), 2.25 (t, J=2.5 Hz, 2H). 13C NMR (CDCl3) δ (ppm): 169.2, 134.2, 133.5, 132.5, 128.4, 128.2, 127.7, 127.6, 127.4, 126.3, 125.8, 79.1, 71.9, 60.3, 40.7, 29.3. The azido compound 112 (1.25 g, 4.28 mmol) was dissolved in DMF (110 mL) and was heated to reflux and left stirring overnight to allow cyclisation. After cooling down the mixture was diluted with EtOAc and was washed with H2O (3 times). The organic phase was dried with MgSO4 and evaporated under reduced pressure to give (R)-8-Naphthalen-2-ylmethyl-4,5,7,8-tetrahydro-1,2,5,8a-tetraaza-azulen-6-one 113 (1.05 g, 84%) as a beige solid. Purity: 95%, LC tR=2.24 min (method A), MS (ESI+): m/z=293 [M+H]+. 1H NMR (CDCl3) δ (ppm): 7.84-7.78 (m, 3H), 7.66 (s, 1H), 7.52-7.45 (m, 3H), 7.27 (dd, J=1.6 and 8.4 Hz, 2H), 6.65 (sl, 1H), 5.21-5.18 (m, 1H), 4.38 (dd, J=5.3 and 17.0 Hz, 1H), 4.27 (dd, J=6.0 and 17.0 Hz, 1H), 3.71 (dd, J=3.3 and 13.6 Hz, 1H), 3.55 (dd, J=8.8 and 13.6 Hz, 1H), 2.98-2.96 (m, 2H). 13C NMR (CDCl3) δ(ppm): 172.3, 133.4, 132.8, 132.7, 132.6, 131.2, 128.9, 128.5, 127.7, 127.6, 127.6, 126.3, 126.0, 58.0, 42.2, 35.5. A 6 M solution of HCl (aq) (10.5 mL) was added to 113 (0.250 g, 0.86 mmol) and splitted in four microwaves tubes. The mixture was heated under microwave conditions at 85° C. for 1 h (Discover-CEM, Method standard: power max 200 W, ramp time 20 min, hold time 60 min, T 85° C., internal pressure max 20 bar). The solution was evaporated under reduced pressure to give compound 114 (322 mg, quant.) as an orange solid. Purity: 97%, LC tR=1.90 min (method A), MS (ESI+): m/z=311 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 8.49 (sl, 2H), 7.87-7.76 (m, 3H), 7.66 (s, 1H), 7.58 (s, 1H), 7.48-7.45 (m, 2H), 7.28 (dd, J=1.6 and 8.4 Hz, 2H), 5.26-5.13 (m, 1H), 3.95 (dd, J=6.3 and 15.8 Hz, 1H), 4.27 (dd, J=5.2 and 15.8 Hz, 1H), 3.44 (dd, J=6.3 and 14.0 Hz, 1H), 3.32 (dd, J=8.6 and 14.0 Hz, 1H), 3.19 (dd, J=9.3 and 17.3 Hz, 1H), 3.06 (dd, J=4.5 and 17.3 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 172.3, 134.8, 133.3, 133.2, 132.4, 132.1, 128.4, 128.2, 128.0, 127.8, 126.7, 126.2, 56.6, 41.4, 31.7.
(R)-4-Fluoro-N-[3-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-3H-[1,2,3]triazol-4-ylmethyl]-benzamide (11). Compound 11 was obtained according to general procedure I from azide 114 and 4-fluoro-benzoyl chloride followed by general procedure D as a white solid (0.012 g, 3 steps 3%). Purity: 97%, LC tR=2.42 min (method A), MS (ESI+): m/z=448 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.21 (s, 1H), 7.86-7.68 (m, 6H), 7.47-7.40 (m, 4H), 7.13 (t, J=8.9 Hz, 3H), 5.33-5.19 (m, 1H), 4.43 (dd, J=6.8 and 15.0 Hz, 1H), 3.86 (dd, J=3.7 and 15.0 Hz, 1H), 3.46-3.43 (m, 2H), 3.13 (dd, J=10.5 and 15.5 Hz, 1H), 2.92 (dd, J=4.1 and 15.5 Hz, 1H). 13C NMR (CD3CN) δ (ppm): 167.0, 165.5, 164.6 (d, J=247 Hz), 146.2, 135.0, 134.7, 133.3, 132.5, 130.2, 129.9, 129.8, 128.0, 127.7, 127.5, 127.4, 127.1, 126.2, 125.8, 115.2 (d, JC-F=21.8 Hz, 2C), 56.9, 41.4, 37.9, 31.3. HRMS m/z calculated for C24H23N5O3F [M+1-1]+ 448.1785, found 448.1788.
(R)—N-[3-(1-Hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-3H-[1,2,3]triazol-4-ylmethyl]-benzamide (12). Compound 12 was obtained according to general procedure I from azide 114 and benzoyl chloride followed by general procedure D as a white solid (0.106 g, 3 steps 24%). Purity: 96%, LC tR=2.39 min (method A), MS (EST+): m/z=430 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 10.58 (s, 1H), 8.80 (m, 2H), 7.84-7.68 (m, 5H), 7.52-7.40 (m, 7H), 7.23 (dd, J=1.6 and 8.4 Hz, 1H), 5.38-5.28 (m, 1H), 4.48 (dd, J=6.4 and 15.5 Hz, 1H), 4.15 (dd, J=4.3 and 15.5 Hz, 1H), 3.42-3.28 (m, 2H), 2.94 (dd, J=9.9 and 15.0 Hz, 1H), 2.65 (dd, J=4.5 and 15.0 Hz, 1H). 13C NMR (DMSO-d6) δ (ppm): 166.5, 166.3, 135.6, 135.0, 134.1, 133.3, 132.9, 132.3, 131.9, 128.7, 128.3, 128.1, 127.9, 127.8, 126.6, 126.1, 56.4, 41.6, 37.9, 31.9. HRMS m/z calculated for C24H24N5O3 [M+H]+ 430.1879, found 430.1880.
(R)-Cyclohexanecarboxylic acid [3-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-3H-[1,2,3]triazol-4-ylmethyl]-amide (13). Compound 13 was obtained according to general procedure I from azide 114 and cyclohexanecarbonyl chloride followed by general procedure D as a white solid (0.041 g, 3 steps 8%). Purity: 98%, LC tR=2.47 min (method A), MS (ESI+): m/z=436 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.29 (s, 0.7H), 7.85-7.75 (m, 3H), 7.49-7.46 (m, 3H), 7.31 (s, 1H), 7.17 (d, J=8.3 Hz, 1H), 7.31 (s, 1H), 6.69 (sl, 1H), 5.23-5.08 (m, 1H), 3.97 (sl, 2H), 3.69-3.47 (m, 1H), 3.43 (d, J=7.3 Hz, 2H), 3.15-2.88 (m, 3H), 1.64 (d, J=12.2 Hz, 5H), 1.29-1.18 (m, 5H). 13C NMR (CD3CN) δ(ppm): 176.3, 166.9, 146.0, 136.0, 134.7, 133.3, 132.3, 131.9, 128.1, 127.8, 127.6, 127.5, 127.2, 126.3, 125.9, 57.1, 44.4, 41.2, 38.0, 31.1, 29.1, 25.5 (2C), 25.3. HRMS m/z calculated for C24H30N5O3 [M+H]′ 436.2349, found 430.2353.
(R)-3-[5-(Acetylamino-methyl)-[1,2,3]triazol-1-yl]-N-hydroxy-4-naphthalen-2-yl-butyramide (24). Compound 24 was obtained according to general procedure I from azide 114 and acetyl chloride followed by general procedure D as a white solid (0.034 g, 3 steps 11%). Purity: 100%, LC tR=2.00 min (method A), MS (ESTE): m/z=368 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.47 (s, 1H), 7.84-7.71 (m, 3H), 7.47-7.45 (m, 3H), 7.31 (s, 1H), 7.15 (d, J=8.4 Hz, 1H), 9.47 (s, 1H), 5.22-5.11 (m, 1H), 4.06 (dd, J=6.3 and 15.6 Hz, 1H), 3.85 (dd, J=4.8 and 15.6 Hz, 1H), 3.41 (d, J=7.8 Hz, 2H), 3.06 (dd, J=10.2 and 15.0 Hz, 1H), 2.88 (dd, J=4.2 and 15.0 Hz, 1H), 1.70 (s, 2.8H). 13C NMR (CD3CN) δ(ppm): 170.2, 167.0, 135.6, 134.7, 133.3, 132.3, 132.1, 128.1, 127.8, 127.5, 127.5, 127.2, 126.2, 125.8, 57.0, 41.4, 38.0, 31.0, 21.8. HRMS m/z calculated for C19H22N5O3 [M+H]+ 368.1723, found 368.1725.
(R)—N-hydroxy-4-naphthalen-2-yl-3-{5-[(3-phenyl-propionylamino)-methyl]-[1,2,3]-triazol-1-yl}-butyramide (25). Compound 25 was obtained according to general procedure I from azide 114 and 3-phenyl-propionyl chloride followed by general procedure D as a white solid (0.030 g, 3 steps 11%). Purity: 100%, LC tR=2.45 min (method A), MS (ESI+): m/z=458 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.53 (s, 1H), 7.82-7.72 (m, 3H), 7.46-7.43 (m, 3H), 7.29-7.10 (m, 7H), 6.94 (m, 1H), 5.13-5.04 (m, 1H), 3.91 (dd, J=5.4 and 15.6 Hz, 1H), 4.15 (dd, J=4.2 and 15.6 Hz, 1H), 3.38 (d, J=7.5 Hz, 2H), 3.09-2.96 (m, 2H), 2.80 (t, J=7.5 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H). 13C NMR (CD3CN) δ (ppm): 172.2, 167.0, 146.0, 141.2, 135.5, 134.6, 133.3, 132.3, 132.0, 128.3, 128.1, 127.8, 127.5 (2C), 127.5, 127.2, 126.3, 126.0, 125.9, 117.3, 57.1, 41.4, 37.9, 37.0, 31.1, 30.9. HRMS m/z calculated for C26H28N5O3 [M+H]+ 458.2192, found 458.2209.
(R,S)—N-hydroxy-3-(5-{[2-(6-methoxy-naphthalen-2-yl)-propionylamino]-methyl}-[1,2,3]triazol-1-yl)-4-naphthalen-2-yl-butyramide (26) Compound 26 was obtained according to general procedure I from azide 114 and 2-(6-methoxy-naphthalen-2-yl)-propionyl chloride followed by general procedure D as a white solid (0.026 g, 3 steps 16%). Purity: 100%, LC tR=2.70 min (method A), MS (ESTE): m/z=538 [M+H]+. 1H NMR (CD3CN) δ (ppm): 9.34 (s, 1H), 7.82-7.79 (m, 1H), 7.71-7.63 (m, 5H), 7.47-7.42 (m, 3H), 7.34 (d, J=8.4 Hz, 1H), 7.18 (s, 2H), 7.09 (dd, J=1.8 and 8.7 Hz, 2H), 7.01 (m, 1H), 5.19-5.08 (m, 1H), 3.93 (d, J=4.8 Hz, 1H), 3.86 (s, 4H), 3.52 (q, J=6.9 Hz, 1H), 3.34 (d, J=6.3 Hz, 2H), 4.15 (dd, J=2.1 and 12.3 Hz, 1H), 2.94 (dd, J=3.3 and 14.7 Hz, 1H), 1.39 (d, J=6.9, 3H). 13C NMR (CD3CN) δ (ppm): 174.2, 166.9, 157.6, 136.9, 135.5, 134.6, 133.7, 133.3, 132.3, 131.9, 129.1, 128.8, 128.0, 127.8, 127.5, 127.5, 127.2, 126.9, 126.3, 126.2, 125.8, 118.7, 117.3, 105.7, 56.9, 55.0, 45.9, 41.3, 37.7, 31.3, 17.6. HRMS m/z calculated for C31H32N5O4 [M+H]+ 538.2454, found 538.2449.
4. Synthesis of Pyrazole
Ethyl 2-[[5-[[(4-fluorobenzoyl)amino]methyl]-1H-pyrazol-3-yl]amino]acetate (132-TVE). To a solution of methyl-2-aminoacetate hydrochloride (6 g, 47.9 mmol, 1.1 equiv) and triethylamine (12.8 mL, 91.4 mmol, 2.1 equiv) in dichloromethane (32 mL) was added dropwise 4-fluorobenzoyl chloride (5.15 mL, 43.5 mmol, 1 equiv). The resulting mixture was stirred at room temperature for 3 h. The organic layer was then washed with HCl (1N), water, brine and was dried over MgSO4. Evaporation of the solvent afforded the desired product which was used without further purification. White/yellow solid (9 g, 98%). Purity: 100%, LC tR=2.47 min, MS (ESI−): m/z=210 [M−H]−. 1H NMR (CDCl3) δ (ppm) 3.77 (s, 3H), 4.19 (d, J=5.4 Hz, 2H), 7.04-7.09 (m, 3H), 7.80-7.84 (m, 2H). 13C NMR (CDCl3) δ (ppm) 41.7, 52.4, 115.6 (JC-F=21.9 Hz), 129.5 (JC-F=8.9 Hz), 129.7 (JC-F=2.9 Hz), 164.9 (JC-F=251.0 Hz), 166.6, 170.6. To a stirred solution of MeCN (2.30 mL, 44.8 mmol, 2.1 equiv) in anhydrous THF (40 mL) under inert atmosphere was added n-BuLi (28.6 mL, 45.8 mmol, 1.6 M in hexane, 2.2 equiv) at −78° C. After the solution was stirred for 30 min at the same temperature, previously obtained methyl 2-[(4-fluorobenzoyl)amino]acetate (4.5 g, 21.3 mmol, 1 equiv) in solution in 20 mL of anhydrous THF was added. The reaction mixture was allowed to warm at room temperature for 3 hours. HCl 1N was added in order to quench the reaction and until the pH of the aqueous layer was equal to 1. The phases were separated. The aqueous layer was extracted three times with EtOAc. The combined organic layers were washed with brine, dried over MgSO4 and the solvent was removed by evaporation. A solution of the residue in ethanol (30 mL) was mixed with hydrazine monohydrate (3.2 mL, 63.9 mmol, 3 equiv) and was refluxed overnight. Ethanol was evaporated and the residue was triturated in water and methanol. The precipitate was then filtered and the desired product was used for the next step without further purification. Brown solid (5 g, 44%). Purity: 95%, MS (ESI+): m/z=235 [M+H]+. 1H NMR (DMSO) δ (ppm) 4.28 (d, J=5.7 Hz, 2H), 4.62 (br s, 2H), 5.26 (s, 1H), 7.24-7.30 (m, 2H), 7.93-7.98 (m, 2H), 8.91 (br s, 1H), 11.25 (br s, 1H). To a solution of N-[β-amino-1H-pyrazol-5-yl)methyl]-4-fluoro-benzamide (2.2 g, 9.39 mmol) in anhydrous methanol (100 mL) were added ethyl glyoxylate (2.23 mL, 11.3 mmol), NaBH3CN (708 mg, 11.3 mmol) and finally glacial acetic acid (0.52 mL, 9.39 mmol). The resulting mixture was heated at 45° C. for 3 h and allowed to cool at room temperature. Ice and water were then added and methanol was evaporated. EtOAc and water were then added, the aqueous layer was extracted three times with EtOAc. The combined organic layer was washed with brine, dried over MgSO4 and the solvent was evaporated. The crude was purified by flash chromatography on silica gel (CH2Cl2/Methanol: 100/0 to 95/5) to give the desired compound as a white solid (790 mg, 26%). Purity: 90%, MS (ESI+): m/z=321 [M+H]+. 1H NMR (DMSO) δ (ppm) 4.28 (d, J=5.7 Hz, 2H), 4.62 (br s, 2H), 5.26 (s, 1H), 7.24-7.30 (m, 2H), 7.93-7.98 (m, 2H), 8.91 (br s, 1H), 11.25 (br s, 1H).
4-fluoro-N-((3-((2-(hydroxyamino)-2-oxoethyl)(naphthalen-2-ylmethyl)amino)-1H-pyrazol-5-yl)methyl)benzamide (53). To a solution of ethyl 2-[[5-[[(4-fluorobenzoyl)amino]methyl]-1H-pyrazol-3-yl]amino]acetate 132 (1 equiv) in anhydrous methanol were added naphthaldehyde, acetic acid and finally NaBH3CN (1.3 equiv). The resulting mixture was refluxed 1 h. NaBH3CN (1.3 equiv) was then added and the mixture was refluxed 1 h30. NaBH3CN (1.3 equiv) was added and the reaction was refluxed 2 h. Water was added to the reaction mixture and methanol was evaporated. Water and EtOAc were then added and the layers were separated. Aqueous layer were extracted (2×) and the combined organic layers were washed with brine and dried over MgSO4. Evaporation of the solvent afforded the crude which was purified by flash chromatography on silica gel (CH2Cl2/MeOH: 99/1 to 95/5) to afford the desired compound (100 mg, 12%). To a solution of hydroxylamine hydrochloride (180 mg, 2.59 mmol, 28 equiv) in dry MeOH (1.2 mL) was added KOH (121 mg, 2.2 mmol, 23 equiv). The mixture was sonicated 5 min at room temperature. Na2SO4 was added in high excess. Filtration of the solids and dropwise addition to a solution of methyl (3R)-3-[4-[[(4-fluorobenzoyl)amino]methyl]triazol-1-yl]-4-(2-naphthyl)butanoate (42 mg, 0.094 mmol, 1 equiv) in dry MeOH (1 mL). The mixture was stirred at 42° C. overnight. KCN (12 mg, 0.188 mmol, 2 equiv) and DIEA (36.5 mg, 0.282 mmol, 3 equiv) were added, and the mixture was stirred to 55° C. for 45 h. The mixture was cooled down to rt, partitioned between EtOAC and water acidified to pH 4 (using aq. saturated NH4C1 and diluted aq. HCl). Extraction with EtOAc, drying with Na2SO4 and evaporation afforded a yellow oily residue as crude product, which was purified by preparative HPLC to give the desired compound as a white solid (17.5 mg, 42%). LC tR: 2.45 min, MS (ESI+) m/z=448 [M+H]+. 1H NMR (MeOD-d4) δ (ppm): 8.42 (s, 1H), 7.88-7.77 (m, 5H), 7.70 (s, 1H), 7.45-7.42 (m, 3H), 7.14 (t, J=17.4 and 8.7 Hz, 2H), 5.70 (s, 1H), 4.69 (s, 2H), 4.49 (s, 2H), 3.71 (s, 2H). 13C NMR (MeOD-d4) δ (ppm): 169.6, 168.8, 136.6, 134.6, 134.0, 131.4, 130.8, 130.7, 129.2, 128.6, 128.5, 127.3, 127.0, 126.8, 126.6, 116.4, 116.1, 90.5, 55.9, 54.6, 53.8, 51.9, 36.4, 30.6. 19F NMR (MeOD) δ (ppm): −109.6. HRMS m/z calculated for C24H23N5O3F [M+H]′ 448.1795, found 448.1785.
5. Synthesis of Oxadiazoles
N—(N-hydroxycarbamimidoylmethyl)-benzamide (135). Compound 134 was obtained according to general procedure S as a white solid (574 mg, 80%). Purity: 93%, LC tR=1.37 min (method A), MS (ESI+): m/z=194 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 9.04 (br s, 1H), 8.70 (br s, 1H), 7.86 (d, J=7.2 Hz, 2H), 7.53-7.46 (m, 3H), 5.38 (br s, 2H), 3.86 (d, J=5.4 Hz, 2H). 13C NMR (DMSO-d6) δ (ppm): 167.2, 151.0, 134.5, 131.8, 128.8, 127.7, 46.1.
4-fluoro-N—(N-hydroxycarbamimidoylmethyl)-benzamide (136). Compound 136 was obtained according to general procedure S as a white solid (2.28 g, 95%). Purity: 100%, LC tR=1.37 min (method A), MS (ESI+): m/z=212 [M+H]+. 1H NMR (DMSO-d6) δ(ppm): 9.05 (br s, 1H), 8.78 (br t, J=6.0 Hz, 1H), 7.97-7.92 (m, 2H), 7.29 (t, J=9.0 Hz, 2H), 5.39 (br s, 2H), 3.85 (d, J=6.0 Hz, 2H). 13C NMR (DMSO-d6) δ (ppm): 166.1, 162.7, 150.9, 131.0, 130.4 (d, JC-F=9 Hz), 115.6 (d, JC-F=22 Hz), 45.9.
2-naphthalen-2-ylmethyl-succinic acid 4-methyl ester (137). To a refluxing suspension of t-BuOK (2.59 g, 23.1 mmol, 1.2 eq) in t-BuOH (17 mL) was carefully added a solution of dimethyl succinate (3.3 mL, 25.0 mmol, 1.3 eq) and 2-naphthaldehyde (3.00 g, 19.2 mmol, 1 eq) in t-BuOH (17 mL). The reaction mixture was stirred at reflux temperature for 3 h, after which the solvent was removed under vacuum. The residue was dissolved in 1M HCl (17 mL) and this solution was extracted with EtOAc (3×50 mL). The organic layers were dried (MgSO4) and concentrated. The resulting monoacid was dissolved and EtOH (24 mL) and aqueous NaOH (2 M, 48 mL) were added. The resultant mixture was stirred under reflux for 16 h, followed by evaporation of the EtOH under reduced pressure. Extra H2O (60 mL) and NaOH (2 M, 10 mL) was added and the mixture was washed with EtOAc (3×60 mL). Next, the aqueous layer was acidified (pH≈1) with 1M HCl, extracted with EtOAc (2×120 mL) and the organic layers were dried (MgSO4) and concentrated. The resulting diacid was dissolved in MeOH (7.4 mL). Amberlyst-15H+ (1.3 g) was added and the reaction mixture was heated under reflux for 16 h. The mixture was filtered over Celite and concentrated under vacuum, resulting in crude ester. The product was purified by flash chromatography (CH2Cl2/MeOH 100:0 to 98:2 (v/v)) to give compound 2-naphthalen-2-ylmethylene-succinic acid 4-methyl ester as a white amorphous solid (2.067 g, 40%). Purity: 75%, LC tR=3.09 min (method A), MS (ESI+): m/z=271 [M+H]+. 1H NMR (CDCl3) δ (ppm): 8.21 (br s, 1H), 7.89-7.86 (m, 4H), 7.58-7.47 (m, 3H), 3.80 (s, 3H), 3.67 (s, 2H). 13C NMR (CDCl3) δ (ppm): 172.8, 171.6, 144.4, 133.4, 133.1, 132.1, 129.3, 128.5, 127.7, 127.2, 126.7, 126.2, 125.3, 52.3, 33.3. Under an atmosphere of argon, Pd/C (0.35 g) and ammonium formate (1.64 g) was added to a solution of the previous acid (0.780 g, 2.89 mmol) in ethanol (29 mL). The mixture was stirred at room temperature for 16 h, then it was filtered over Celite. Solvent was removed under vacuum. The residue was solubilized in water and acidified (pH≈1) with 1M HCl, extracted with CH2Cl2 (2×120 mL) and the collected organic layers were dried (MgSO4) and concentrated. The residue was purified by flash chromatography (CH2Cl2/MeOH 100:0 to 98:2 (v/v)) to give compound 111 as a yellow oil (0.663 g, 84%). Purity: 84%, LC tR=2.65 min (method A), MS (ESI−): m/z=271 [M−H]−. 1H NMR (MeOD) δ (ppm): 9.74 (br s, 1H, OH), 7.89-7.84 (m, 3H), 7.70 (s, 1H), 7.54-7.48 (m, 2H), 7.39 (d, J=8.4 Hz, 1H), 3.66 (s, 3H), 3.37-3.32 (m, 2H), 3.00 (dd, J=10.5 and 15.3 Hz, 1H), 2.75 (dd, J=8.7 and 17.1 Hz, 1H), 2.51 (dd, J=4.5 and 17.1 Hz, 1H). 13C NMR (MeOD) δ (ppm): 180.1, 172.2, 135.7, 133.5, 132.4, 128.3, 127.7, 127.6, 127.6, 127.2, 126.2, 125.7, 42.9, 37.5, 34.6.
2-Naphthalen-2-ylmethyl-malonic acid monomethyl ester (138). To a solution of naphthaldehyde (3.5 g, 22.4 mmol) in DMSO (10 mL) was added proline (253 mg, 3.2 mmol) and the mixture was stirred 15 min at room temperature. Dimethylmalonate (5.1 mL, 44.8 mmol) was then added and the reaction media was stirred for 24 hours. EtOAc was added and the organic layer was washed with water, brine and was dried over MgSO4. Evaporation of the solvent afforded the crude which was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 100:0 to 90:10). Viscous yellow oil (1.6 g, 26%). Purity: 100%, LC tR=3.40 min, MS (ESI+): m/z=271 [M+H]+. 1H NMR (CDCl3) δ (ppm): 3.87 (s, 6H), 7.45-7.56 (m, 3H), 7.80-7.85 (m, 3H), 7.93 (s, 2H). 13C NMR (CDCl3) δ (ppm): 52.7, 125.0, 125.5, 126.8, 127.7, 127.8, 128.7, 128.8, 130.3, 131.1, 133.1, 134.1, 143.1, 164.6, 167.3. To a solution of 2-Naphthalen-2-ylmethylene-malonic acid dimethyl ester (1.52 g, 5.64 mmol) in degazed EtOH (100 mL) were added ammonium formate (3.2 g, 50.8 mmol) and palladium (10%) on charcoal (600 mg, 0.56 mmol). The reaction was stirred for 4 hours and the media was then filtered over celite. The residue was dissolved in EtOAc, washed with water and brine and finally dried over MgSO4. Evaporation of the solvent afforded the desired product which was used without further purification. Yellow oil (1.53 g, Quant.). Purity: 100%, LC tR=3.37 min, MS (ESI+): m/z=273 [M+H]+. 1H NMR (CDCl3) δ (ppm): 3.42 (d, J=7.8 Hz, 2H), 3.70 (s, 6H), 3.82 (t, J=7.8 Hz, 1H), 7.34 (dd, J=1.5 Hz and 8.4 Hz, 1H), 7.42-7.49 (m, 2H), 7.67 (s, 1H), 7.77-7.82 (m, 3H). 13C NMR (CDCl3) δ (ppm): 35.0, 52.6, 53.6, 125.7, 126.1, 127.0, 127.4, 127.7, 128.3, 132.4, 133.5, 135.3, 169.3. NaOH (259 mg, 6.46 mmol) was added to a solution of 2-Naphthalen-2-ylmethyl-malonic acid dimethyl ester (1.53 g, 5.62 mmol) in MeOH (10 mL). The reaction was stirred overnight and the solvent was removed. The residue was dissolved in aqueous NaHCO3 and washed twice with CH2Cl2. The aqueous layer was acidified with fuming HCl and then extracted with EtOAc. The combined organic layer was dried over MgSO4 and the solvent was evaporated to afford the desired product as a white solid (1.45 g, Quant.). Purity: 100%, LC tR=2.77 min. 1H NMR (CDCl3) δ (ppm): 3.41 (dd, J=2.7 and 7.8 Hz, 2H), 3.70 (s, 3H), 3.83 (t, J=7.8 Hz, 1H), 7.33 (dd, J=1.8 and 8.4 Hz, 1H), 7.42-7.50 (m, 2H), 7.67 (s, 1H), 7.76-7.82 (m, 3H), 8.70 (br s, 1H)
N-[5-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzamide (28). Compound 28 was obtained according to general procedure T from 135 and 138 followed by general procedure P followed by general procedure D as a colorless oil (65 mg, 41%). Purity: 93%, LC tR=2.87 min (method A), MS (ESI+): m/z=431 [M+H]+. 1H NMR (acetone-d6) δ (ppm): 8.29 (br s, 1H), 7.95-7.75 (m, 5H), 7.61 (s, 1H), 7.55-7.42 (m, 5H), 7.29-7.27 (m, 1H), 4.66 (s, 2H), 3.96-3.93 (m, 1H), 3.24 (d, J=6.6 Hz, 2H), 2.66-2.62 (m, 2H). 13C NMR (acetone-d6) δ (ppm): 181.5, 168.2, 167.0, 166.6, 135.5, 134.3, 133.6, 132.4, 131.4, 128.4 (2C), 128.0, 127.6 (2C), 127.5, 127.3 (3C), 126.0, 125.6, 38.8, 36.4, 35.1, 34.6. HRMS m/z calculated for C24H23N4O4 [M+H]+431.1719, found 431.1727.
4-fluoro-N-[5-(1-hydroxycarbamoylmethyl-2-naphthalen-2-yl-ethyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzamide (27). Compound 27 was obtained according to general procedure T from 136 and 138 followed by general procedure P followed by general procedure D as a colorless oil (44 mg, 28%). Purity: 100%, LC tR=2.90 min (method A), MS (ESI+): m/z=449 [M+H]+. 1H NMR (acetone-d6) δ (ppm): 8.35 (br t, 1H, NH), 8.02-7.98 (m, 2H), 7.82-7.74 (m, 3H), 7.60 (s, 1H), 7.46-7.40 (m, 2H), 7.29-7.20 (m, 3H), 4.65 (d, J=6.0 Hz, 2H), 4.00-3.90 (m, 1H), 3.24 (d, J=7.2 Hz, 2H), 2.65 (t, J=8.7 Hz, 2H). 13C NMR (acetone-d6) δ (ppm): 181.5, 168.1, 167.1, 165.7, 164.6 (d, JC-F=248 Hz), 163.7, 135.5, 133.5, 132.4, 130.7, 130.0, 129.9, 128.0, 127.6 (2C), 127.5, 127.3, 126.0, 125.6, 115.2 (d, JC-F=22 Hz), 38.8, 36.4, 35.1, 34.6. HRMS m/z calculated for C24H22N4O4F [M+H]+449.1625, found 449.1668.
4-Fluoro-N-[5-(1-hydroxycarbamoyl-2-naphthalen-2-yl-ethyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzamide (44). To a solution of the carboxylic acid oxadiazole (1 equiv), obtained according to general procedure T from 136 and 138 followed by general procedure P, in DMF were added HOBt (5 equiv), EDCI (4 equiv), and N-methylmorpholine (12 equiv) and the reaction media was stirred 15 min at room temperature. O-(tert-butyldimethylsilyl)hydroxylamine (3 equiv) was then added and the resulting mixture was stirred overnight. DMF was evaporated and the residue was dissolved in EtOAc, and washed with water and brine. The organic layer was then dried over MgSO4 and the solvent was evaporated. Purification by flash chromatography to afford a yellow solid (20 mg, 2%). Purity: 96%, LC tR=2.85 min, MS (ESI+): m/z=435 [M+H]+. 1H NMR (DMSO-d6) δ (ppm): 3.46 (d, J=7.5 Hz, 2H), 4.24 (t, J=7.8 Hz, 1H), 4.57 (d, J=6.0 Hz, 2H), 7.31-7.37 (m, 2H), 7.39-7.42 (m, 1H), 7.45-7.49 (m, 2H), 7.72 (s, 1H), 7.77-7.88 (m, 3H), 7.94-7.99 (m, 2H), 9.13 (s, 1H), 9.22 (t, J=5.7 Hz, 1H), 10.94 (s, 1H). 13C NMR (DMSO-d6) δ (ppm) 35.2, 35.5, 44.1, 115.9 (JC-F=22 Hz), 120.0, 126.1, 126.6, 127.8, 127.9, 128.3, 130.4 (JC-F=9 Hz), 130.6, 132.4, 133.4, 135.6, 163.7, 164.5 (JC-F=247 Hz), 165.7, 168.8, 177.6. HRMS m/z calculated for C23H20N4O4F [M+H]+435.1469, found 435.1438.
1. Cell Culture
(a) OPM2 Cell Culture
Human multiple myeloma cells OPM2 (ACC 50, DSMZ) are maintained at 0.3-0.7 106 cells/mL in RPMI1640 medium supplemented with GlutaMAX™, 10% fetal bovine serum and 1% penicillin-streptomycin.
(b) HeLa Cell Culture
Human HeLa cells (uterine cervical carcinoma) are maintained in DMEM medium supplemented with GlutaMAX™, 10% fetal bovine serum and 1% penicillin-streptomycin.
(c) MCF7 Cell Culture
Human epithelial MCF7 cells (mammary gland adenocarcinoma) are maintained in EMEM medium supplemented with GlutaMAX™, 10% fetal bovine serum, 0.01 mg insulin and 1% penicillin-streptomycin.
(d) RPMI 8226 and MOPC315.BM Cell Culture
RPMI 8226 and MOPC315.BM cells were cultured in RPMI 1640 medium supplemented with L-glutamine, non-essential amino acids, 2-mercaptoethanol and 10% fetal calf serum.
(e) TLM1 Cell Culture
Canine melanoma TLM-1 cells are maintained in DMEM medium supplemented with GlutaMAX™, 10% fetal bovine serum, 1× Hepes and 1% penicillin-streptomycin.
2. High Content Screening to Identify Compounds that Sensitize Cells to RE stressors and boost apoptosis.
We developed a cell-based, high content assay allowing the rapid detection and quantification of apoptotic cells. Caspase-3/7 activity was detected in real-time using the NucView™ 488 Caspase-3 substrate (Biotium), where the caspase-3/7 substrate peptide sequence DEVD is attached to a nucleic acid dye. The NucView™ 488 reagent crosses the plasma membrane to enter the cytoplasm, where the dye is unable to bind to DNA and remains non-fluorescent. Once the substrate has been cleaved by activated caspase-3/7 in apoptotic cells, the high-affinity DNA dye is released and migrates to the cell nucleus to stain the nucleus with a bright green fluorescence. All cell nuclei were also labeled with Hoechst 33342 (Ozyme).
For OPM2, briefly, nanoliters of compound and cytotoxic agent in DMSO stock solutions were first transferred and combined in 384-microtiter well plate using an Echo555 liquid handler (Labcyte). The exact volume of each solution to be transferred was calculated to achieve subsequently the target concentration in the 70 μL volume incubate. Final DMSO content was maintained at 0.2%. Then, 70 μL of cells treated with 1 μM NucView™ 488 and 20 ng/mL Hoechst 33342 were distributed to the plates to obtain a final density of 12,000 cells per well. The microplates were incubated at 37° C. and 5% CO2 in an automated LiCONiC STR240 UB cell incubator, and images were acquired at various times at 405 nm and 488 nm with the high-resolution automated confocal microscope INCell 6000 (GE). Four fields per well were read with the 20× objective in a confocal mode. The BioCel workstation was managed by VWorks automation control software
(Agilent).
For HeLa and MCF7 cells, briefly, cells were seeded in 384-microtiter well plate (1500 HeLa cells or 5000 MCF7 cells in 40 μL full medium per well) and let to attach overnight. Then the compound of the invention and the RE stressor were transferred to the cell culture plate using an Echo555 liquid handler (Labcyte) and 30 μL medium containing NucView™ and Hoechst 33342 were added. Final NucView™ 488 and Hoechst 33342 concentrations were 1 μM and 20 ng/mL, respectively. DMSO content was maintained at 0.2%. The microplates were incubated at 37° C. and 5% CO2 in an automated LiCONiC STR240 UB cell incubator, and images were acquired at various times at 405 nm and 488 nm with the high-resolution automated confocal microscope INCell 6000 (GE). Four fields per well were read with the 20× objective in a confocal mode. The BioCel workstation was managed by VWorks automation control software (Agilent).
For all cells, Columbus software (Perkin) was used for image quantification and analysis. First, Hoechst 33342 staining was used for an automated detection of nuclei, in which the mean fluorescence intensity for NucView™ 488 was quantified. Inspection of control cells at initial time was used to set a threshold level of NucView™ 488 fluorescence to separate non apoptotic cells from apoptotic cells. Finally, for each well, the result was expressed as percentage of apoptotic cells in the total cell population. CC50 values were calculated from concentration-response curves by a nonlinear regression analysis at four parameters using Excel Fit or GraphPad Prim softwares.
Activities were Calculated as Either:
Activity=(% of apoptotic cells with cytotoxic agent in combination with compound of the invention)/(% of apoptotic cells with cytotoxic agent alone).
Activity=(CC50 of agent alone)/(CC50 of agent with compound).
3. Effect of the Combination of Epoxomicin and Compound (3) of the Invention on Murine and Human Myelomas.
Cells were seeded in 24-well culture plates at 150,000 cells/well/mL. Compounds (EPX and 3) were dissolved and added to culture medium at a final concentration of 1%. Cells were treated with protease inhibitors for 24 and 48 hours before analysis for apoptosis and cell death by flow cytometry. The cells were centrifuged at 300×g for 5 minutes and washed with cold FACS buffer (PBS 1× with 2 mM EDTA and 0.5% BSA. Then the cells were resuspended in Annexin V binding buffer (BioLegend) at a concentration of 106 cells/mL, and 100 μL of each sample was transferred to a 5 mL FACS tube. 5 μL of Annexin V-FITC Ab (BioLegend) and 5 μL of propidium iodide (BioLegend) were added to each sample and incubated 15 min at RT. Finally, 400 μL of Annexin V binding buffer was added to each tube and the cells were analyzed using BD FACSCanto or BD Fortessa machines within an hour. Annexin+ but PI− cells were identified as apoptotic and Annexin+ PI+ as dead cells.
4. Effect of the Combination of X-Ray Radiation and Compounds of the Invention on Canine Melanoma TLM1 Cell Line
TLM-1 cells were seeded with the BioTek MicroFlo™ dispenser in μClear® black 384-well plates (Greiner) at the density of 20,000 cells/mL in 50 μL. Compounds for pre-treatment were added 2 h after the seeding, using an Echo 550 (Labcyte). The next day, cells were washed five times with 40 μL media without phenol red with a plate washer (Hydrospeed, Tecan). Cells were irradiated with 4, 6, 8, 12, 16, 20 and 24 Gray (Gy). Around 1.5 h after irradiation, 20 μL of media containing compounds at the desired concentration, Hoechst 33342 (40 ng/mL, Thermofisher) for nuclei staining and propidium iodide (1 μg/mL, Thermofisher) for labeling dead cells were added to the cell plates with a Bravo liquid handler (Agilent). This step was repeated every 72 h after radiation. Plates were incubated at 37° C. and 5% CO2. Images were acquired at various times with DAPI filter set (ex.405/em.455 nm) and Texas Red filter set (ex.561/em.605 nm) with the high-resolution automated confocal microscope INCell 6000 (GE) in 20× magnification in non-confocal mode. 4 images per well were collected. Images were analyzed with Columbus software (Perkin Elmer). In this system we used predefined building blocks to measure different parameters on nuclei detected. We applied a threshold based on propidium iodide intensity in nuclei of controls to select and count dead cells. By dividing the number of dead cells by the number of nuclei detected we obtained the percentage of dead cells. AUC were determined in one experiment with triplicate incubates on percentage of cells labelled with propidium iodide and were corrected by the percentage of cells labelled with propidium iodide at 0 Gy. Ratio of AUC corrected was determined as followed: Activity=(Area of the dose-concentration curve of cytotoxic agent combined with one concentration of compound corrected by the effect of the compound alone at 0Gy)/(Area of the dose-concentration curve of radiation alone corrected by the effect of the vehicle at 0Gy).
5. Effect of the Combination of UV-C Radiation and Compounds of the Invention on Canine Melanoma TLM1 Cell Line
TLM-1 cells were seeded in μClear® black 384-well plates (Greiner) at the density of 60,000 cells/mL in 50 Compounds were added 24 h after the seeding, using an acoustic nanodispensor Echo 550 (Labcyte). The next day, cells were washed five times with 40 μL phosphate buffer saline (PBS) with a plate washer (Hydrospeed, Tecan). 10 μL of PBS was left per well. Plate was submitted to UV-C(Stratalinker® UV Crosslinker, Artisan Technologies Group) with different doses (J/m2). After UV-C radiation, compounds were added using an acoustic nanodispensor Echo 550 (Labcyte). We complete to 50 μL of media per well. Plates were incubated at 37° C. and 5% CO2. Some hours before image acquisition, propidium iodide (1 Thermofisher) for labeling dead cells and Hoechst 33342 (80 ng/mL, Thermofisher) for nuclei staining were added using an acoustic nanodispensor Echo 550 (Labcyte). Images were acquired at various times with DAPI filter set (ex.405/em.455 nm) and Texas Red filter set (ex.561/em.605 nm) with the high-resolution automated confocal microscope INCell 6000 (GE) in 20× magnification in non-confocal mode. 4 images per well were collected. Images were analyzed with Columbus software (Perkin Elmer). In this system we used predefined building blocks to measure different parameters on nuclei detected. We applied a threshold based on propidium iodide intensity in nuclei of controls to select and count dead cells. By dividing the number of dead cells by the number of nuclei detected we obtained the percentage of dead cells. Activity refers to the ratio of the percentage of cells labelling with propidium iodide with pre-post treatment with the compound on the percentage of cells labelling with propidium iodide with pre-post treatment with vehicle, 48 h after the UV-C radiations.
Compounds of the invention increase proteasome inhibitor cytotoxicity in human multiple myeloma cell lines: for example, Table 2 shows activities of compounds of the invention in combination with carfilzomib (CFZ) on OPM2 cells. Results show that activities of compounds of the invention in combination with CFZ on OMP2 cells are higher compared to the activity of CFZ alone.
a% OPM2 apoptotic cells of (CFZ + compound of the invention) vs. CFZ alone
Compounds of the invention increase cytotoxicity of tunicamycin (TUN) in human multiple myeloma cell line (OPM2) (Table 3). Results show that activities of compounds of the invention in combination with TUN on OMP2 cells are higher compared to the activity of TUN alone.
a% OPM2 apoptotic cells of (TUN + compound of the invention) vs. TUN alone
Compounds of the invention increase cytotoxicity of epoxomycin (EPX) in multiple myeloma cell lines (RPMI and MOPC315.BM) (
Compounds of the invention boost proteasome inhibitor cytotoxicity in cells other than multiple myeloma cell lines: for example, Table 4 shows activities of compounds of the invention in combination with CFZ on cervical cancer cell lines (Hela) (Table 4). Results show that activities of compounds of the invention in combination with CFZ on HeLa cells are higher compared to the activity of CFZ alone.
a% Hela apoptotic cells of (CFZ + compound of the invention) vs. CFZ alone
Compounds of the invention increase the potency of proteasome inhibitors such as carfilzomib: lower doses of carfilzomib can be used to reach the same effect when used in combination with compounds of the invention. As shown in Table 5 & Table 6, the ratios (CC50 of proteasome inhibitor alone)/(CC50 of proteasome inhibitor in combination with compound of the invention) is higher than 1.
aratio (CC50 of CFZ alone)/(CC50 of CFZ in combination with compound of the invention (7.5 μM)).
bcalculated with compound at a concentration of 3.7 μM.
aratio (CC50 of CFZ alone)/(CC50 of CFZ in combination with compound of the invention (7.5 μM));
bratio (CC50 of CFZ alone)/(CC50 of CFZ in combination with compound of the invention (15 μM)).
Compounds of the invention increase the potency of other cytotoxic agents, in particular
HDAC inhibitors such as Panobinostat (PAN) and proteasome inhibitors such as Nelfinavir (NEL): lower doses of cytotoxic agents can be used to reach the same effect when used in combination with compound 3 of the invention (Table 7 & Table 8).
aratio of AUC of cytotoxity of PAN with and without 15 μM (3).
aratio of AUC of cytotoxity of NEL with and without 15 μM (3).
Compounds 3, 80, 93, 144, 168, 169, 186, 187 of the invention increase the potency of X-ray radiations: lower doses of X-ray radiations can be used to reach the same effect when used in combination with compounds 3, 80, 93, 144, 168, 169, 186, or 187 of the invention (Table 9).
aratio of (corrected AUC of X-ray radiation in combination with compound of the invention (30 μM)/corrected AUC of X-ray radiation alone)
Compounds 3 and 80 of the invention increase the potency of UV-C radiations: lower doses of UV-C radiations can be used to reach the same effect when used in combination with compounds 3 or 80 of the invention (Table 10).
aTLM-1 cells were treated with 15 μM of compound of the invention 24 h before the UV-C radiation and just after UV-C radiation. Activity refers to the ratio of the percentage of cells labelling with propidium iodide with pre-post treatment with the compound at 15 μM on the percentage of cells labelling with propidium iodide with pre-post treatment with vehicle, 48 h after the UV-C radiations at 400 J/m2. Results are the mean of ratio obtained in three independent experiments.
| Number | Date | Country | Kind |
|---|---|---|---|
| 19305058.0 | Jan 2019 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2020/051058 | 1/16/2020 | WO | 00 |
