Antiviral heterocyclic compounds

Abstract

The present invention relates to a compound of formula (I): [Formula 1] Disclosed herein are compounds of TIFF2024507561000648.tif2138 or pharma- ceutically acceptable salts, esters, or prodrugs thereof, which (I) inhibit an inhibitor of human respiratory syncytial virus (HRSV) or human metapneumovirus (HMPV). The invention further relates to a pharmaceutical composition comprising the above-mentioned compound for administration to a subject suffering from a HRSV or HMPV infection. The invention also relates to a method of treating a HRSV or HMPV infection in a subject by administering a pharmaceutical composition comprising a compound of the invention.

Description

[Technical Field]

[0001] Related applications This application claims the benefits of U.S. Provisional Patent Application No. 63 / 154,318 filed on 26 February 2021, U.S. Provisional Patent Application No. 63 / 168,705 filed on 31 March 2021, U.S. Provisional Patent Application No. 63 / 171,895 filed on 07 April 2021, and U.S. Provisional Patent Application No. 63 / 293,339 filed on 23 December 2021. All teachings of the above applications are incorporated herein by reference.

[0002] The present invention relates to compounds and pharmaceutical compositions that are generally useful as inhibitors of respiratory syncytial virus (RSV) and human metapneumovirus (HMPV). [Background technology]

[0003] Human respiratory syncytial virus (HRSV) is a negative sense virus containing an unsegmented single-stranded linear RNA genome. As a paramyxovirus of the genus Pneumoviridae with two serotypes, HRSV contains 10 genes encoding 11 proteins. The nucleocapsid protein (N), RNA polymerase protein (L), phosphorylated protein (P), and transcriptional anti-termination factor (M2-1) together with the RNA genome form the ribonucleoprotein (RNP) complex. Several small molecule compounds have been shown to target the RNP complex. Furthermore, the fusion protein (F), which is crucial for viral attachment to the host, has been widely studied. High-resolution structures of the F protein that interact with inhibitors have been achieved, but structural studies using the N protein are in the early stages of development. HRSV protein research and its direct results, the F, L, and N proteins, have been a major focus of drug discovery efforts.

[0004] The increasing effort in HRSV drug discovery is due to HRSV being the leading cause of acute lower respiratory tract infections (ALRIs) in patients of all ages. In addition to respiratory infections, high-risk patient populations during HRSV infection include the elderly, immunocompromised individuals, children up to 2 years of age, and patients with chronic obstructive pulmonary disease (COPD) or chronic heart failure (CHF). Over a four-year period, HRSV has been found to cause 177,500 hospitalizations and 14,000 deaths in the elderly population in the United States. It is well known that almost all children are infected with HRSV in the first three years of life, and that HRSV infection is more severe in premature infants. In fact, HRSV is the most common cause of bronchiolitis and pneumonia in infants under 1 year of age in the United States. It is estimated that approximately 3.2 million hospitalizations and 66,000 deaths worldwide in children under 5 years of age are due to HRSV. HRSV is associated with a higher rate of infant deaths and hospitalizations in infants under 1 year of age than influenza.

[0005] HRSV infection can also affect healthy individuals, and recurrent HRSV infections can occur over a period of up to two months. Symptoms are similar to a common cold in healthy individuals, but fever, wheezing, rapid and difficult breathing, and cyanosis may occur in more severe cases.

[0006] Currently, treatment options for HRSV infection are very limited, and there is no vaccine due to the unsuccessful nature of previous attempts. Palivizumab is a monoclonal antibody approved for prophylactic use, but its high cost limits its use. Palivizumab is generally used only in high-risk infants, such as premature infants or those with heart / lung disease, but it has only been effective in reducing hospitalizations in 60% of cases. Ribavirin is approved as an inhalation treatment option, but its effectiveness is limited, and there are associated safety concerns. Given the treatment options and the consistent seasonality of HRSV outbreaks, the development of new therapeutic agents for HRSV is desirable.

[0007] Several RSV fusion inhibitors are disclosed in the following publications: International Publication No. 2010 / 103306, International Publication No. 2012 / 068622, International Publication No. 2013 / 096681, International Publication No. 2014 / 060411, International Publication No. 2013 / 186995, International Publication No. 2013 / 186334, International Publication No. 2013 / 186332, International Publication No. 2012080451, International Publication No. 2012 / 080 Issue 450, International Publication No. 2012 / 080449, International Publication No. 2012 / 080447, International Publication No. 2012 / 080446, International Publication No. 2015 / 110446, International Publication No. 2017 / 009316, J.Med.Chem.2015,58,1630-1643, Bioorg.Med.Chem.Lett.,2015,25,976-981, Reserve Nat.Commun.,2017,8,167. Examples of other N protein inhibitors for the treatment of HRSV are disclosed in the following publications: International Publication 2004 / 026843, J.Med.Chem. 2006, 49, 2311-2319, and J.Med.Chem. 2007, 50, 1685-1692. Examples of L protein inhibitors for HRSV are disclosed in the following publications: International Publication 2011 / 005842, International Publication 2005 / 042530, Antiviral Res. 2005, 65, 125-131, and Bioorg.Med.Chem.Lett. 2013, 23, 6789-6793. Examples of nucleoside / polymerase inhibitors are disclosed in the following publications: International Publication No. 2011 / 005842, International Publication No. 2013 / 242525, International Publication No. 2014 / 031784, International Publication No. 2015 / 026792, International Publication No. 2016 / 0055791, International Publication No. 2016 / 138158, and J.Med.Chem. 2015, 58, 1862-1878.

[0008] Similarly, human metapneumovirus (HMPV), a negative-sense single-stranded RNA enveloped virus belonging to the Pneumoviridae family and Metapneumovirus genus, discovered by van Den Hoogen in 2001, is also a common cause of acute lower respiratory tract infections (ALRTIs). Although often mild, this virus can be serious and life-threatening in high-risk groups such as children under 5 years of age, older adults over 65 years of age, and adults with underlying medical conditions (e.g., chronic obstructive pulmonary disease (COPD), asthma, congestive heart failure, or diabetes). In healthy adults over 65 years of age, the annual incidence of HMPV infection is 1.2 / 1,000, but 38% of individuals have underlying medical conditions (e.g., COPD), and the likelihood of symptomatic disease and need for medical attention is twice as high. In immunocompromised individuals, HMPV accounts for 6% of all respiratory infections in lung transplants and 3% of lower respiratory tract infections associated with stem cell transplants. HMPV infection is also thought to be associated with acute graft rejection.

[0009] Similar to HRSV, infection is thought to occur via glycoprotein (G) protein interactions, followed by fusion via the F protein. The HMPV L protein sequence is homologous to the HRSV L protein.

[0010] HMPV infection is the second most common cause of lower respiratory tract infections in children (after HRSV) and is also a concern in older populations. There are four subtypes of HMPV found in clinical isolates (A1, A2, B1, and B2). Reinfection can occur throughout childhood after the initial infection. There are currently no available treatments for HMPV infection.

[0011] Given the seasonality and predictability of HRSV and HMPV outbreaks, the prevalence of HRSV in elderly care facilities, and the severity of infections in high-risk infants, the need for potent and effective treatment of HRSV and HMPV is clear. This invention identifies compounds that are potent heterocyclic molecules against HRSV-A / B and HMPV. This invention includes methods for preparing these molecules, methods for assays based on RSV cells, HMPV-GFP cells, and HMPV-TN / 1501 / A1 cells, and small molecules with potential to treat HRSV / HMPV infections. [Overview of the Initiative]

[0012] The present invention provides compounds represented by formula (I) that can be used to treat or prevent viral infections (particularly HRSV or HMPV), as well as pharmaceutically acceptable salts, esters, and prodrugs thereof. [ka] During the ceremony, A is 1) Arbitrarily substituted aryls; and 2) Selected from the group consisting of arbitrarily substituted heteroaryls, R1 and R2 are independent of each other as follows: 1) Hydrogen; 2) Fluorine; and 3) Selected from the group consisting of arbitrarily substituted -C1~C6 alkyl groups, Alternatively, R1 and R2, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 6-membered rings. Z is 1) Hydrogen; 2) Halogens; and 3) Selected from the group consisting of arbitrarily substituted -C1~C6 alkyl groups, W is 1) Hydrogen; 2) Arbitrarily substituted -C1~C6 alkoxys; 3) Any substituted -C1~C6 alkyl; and 4) selected from the group consisting of optionally substituted -C3 - C6 cycloalkyl, G is, 1) -C(O)OR ; 2) -C(O)NR 11 R 12 ; 3) optionally substituted -C1 - C6 alkyl -CN; 4) optionally substituted -C1 - C6 alkyl -C(O)NR 11 R 12 ; 5) optionally substituted -C1 - C6 alkyl -C(O)NR 11 S(O)2R 12 ; 6) optionally substituted -C1 - C6 alkyl -OC(O)NR 11 R 12 ; 7) optionally substituted -C1 - C6 alkyl -NHR 13 ; 8) optionally substituted -C1 - C6 alkyl -NHC(O)R 13 ; n is 1, 2 or 3, preferably, n is 1 or 2, Y is O, S(O)2, or NR 14 ; E is, 1) optionally substituted aryl;​​​​​​​​​​​​​​​​​​​​​​​​​2) Arbitrarily substituted -C1~C8 alkyl groups; 3) Arbitrarily substituted -C3~C8 cycloalkyl groups; 4) Arbitrarily substituted 4- to 8-membered complex rings; 5) Arbitrarily substituted aryl; 6) Arbitrarily substituted arylalkyls; 7) Any substituted heteroaryls; and 8) Selected from the group consisting of arbitrarily substituted heteroarylalkyls, R 12 Each occurrence independently of the following: 1) Hydrogen; 2) Arbitrarily substituted -C1~C8 alkyl groups; 3) Arbitrarily substituted -C3~C8 cycloalkyl groups; 4) Arbitrarily substituted 4- to 8-membered complex rings; 5) Arbitrarily substituted aryl; 6) Arbitrarily substituted arylalkyls; 7) Any substituted heteroaryls; and 8) Selected from the group consisting of arbitrarily substituted heteroarylalkyls, Alternatively, R 11 and R 12 These, together with the nitrogen atoms to which they are bonded, form a 3- to 12-membered heterocyclic ring, preferably, the 3- to 12-membered heterocyclic ring is, but is not limited to, morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, and azetidine. R 13 Each occurrence independently of the following: 1) Arbitrarily substituted -C1~C8 alkyl groups; 2) Optionally substituted -C3~C8 cycloalkyl groups; 3) Arbitrarily substituted 4- to 8-membered complex rings; 4) Arbitrarily substituted aryl; 5) Arbitrarily substituted arylalkyls; 6) Any substituted heteroaryls; and 7) Selected from the group consisting of arbitrarily substituted heteroarylalkyls, R 14teeth, 1) Hydrogen; 2) Arbitrarily substituted -C1~C8-alkyl; and 3) Selected from optionally substituted -C3~C8-cycloalkyl groups, each of the above preferred groups can be used in combination with one, any, or all other preferred groups. [Modes for carrying out the invention]

[0013] One embodiment of the present invention is the compound of formula (I) above or a pharmaceutically acceptable salt thereof.

[0014] In certain embodiments of the compound of formula (I), Y is O.

[0015] In certain embodiments of the compound of formula (I), Y is O and n is 1 or 2.

[0016] In certain embodiments of the compound of formula (I), R1 is hydrogen or F.

[0017] In certain embodiments of the compound of formula (I), R2 is hydrogen or fluorine.

[0018] In certain embodiments of the compound of formula (I), Z is hydrogen, Cl, or F.

[0019] In certain embodiments of the compound of formula (I), R1 is hydrogen, R2 is hydrogen, and Z is hydrogen.

[0020] In certain embodiments of the compound of formula (I), W is optionally substituted methyl, optionally substituted ethyl, or optionally substituted cyclopropyl.

[0021] In certain embodiments of the compound of formula (I), W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3.

[0022] In certain embodiments of the compound of formula (I), R3 is -OH.

[0023] In certain embodiments of the compound of formula (I), R4 is optionally substituted methyl or optionally substituted cyclopropyl.

[0024] In certain embodiments of the compound of formula (I), R3 is OH and R4 is CF3 or cyclopropyl.

[0025] In certain embodiments of the compound of formula (I), R1 is hydrogen, R2 is hydrogen, R3 is OH, and R4 is CF3.

[0026] In certain embodiments of the compound of formula (I), G is -C(O)NR 11 R 12 In certain embodiments, G is -C(O)NH2.

[0027] In certain embodiments of the compound of formula (I), G is -CH2NHR 13 -CH2C(O)NR 11 R 12 -CH2NHC(O)R 13 -CH2OC(O)NR 11 R 12 -CH2CN, or -CH2C(O)NR 11 S(O)2R 12 In certain embodiments, G is -CH2C(O)NH2.

[0028] In certain embodiments of the compound of formula (I), A is obtained by removing a hydrogen atom as follows: [ka] One of these is selected, and each of these bases is optionally substituted.

[0029] In certain embodiments of the compound of formula (I), A is as follows: [ka] The groups are selected from those shown, and each of these groups is optionally substituted.

[0030] In a particular embodiment of the compound of formula (I), A is: [ka] Selected from the bases shown.

[0031] In a particular embodiment of the compound of formula (I), A is [ka] or The image is JPEG0007871278000006.jpg1517, where Ra is hydrogen, halogen, -CN, NO2, -OR 11 , -NR 11 R 12 , -NR 11 C(O)R 12 , -NR 11 S(O)2R 12 -S(O)2R 12 -S(O)2NR 11 R 12 , -NR 11 C(O)NR 11 R 12 , -C(O)R 11 , -C(O)OR 11 -C(O)NR 11 R 12 Rb and Rb' are, optionally substituted -C1~C6 alkyl, optionally substituted -C3~C8-cycloalkyl, optionally substituted 3-membered to 8-membered heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl, and Rb and Rb' are each independently hydrogen, halogen, or -OR 11 , -NR 11 R 12 The following are selected: optionally substituted -C1-C6-alkyl, optionally substituted -C3-C8-cycloalkyl, optionally substituted 3- to 8-membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl. Alternatively, Rb and Rb' together with the carbon atoms to which they are bonded form a 4- to 7-membered ring fused with the phenyl ring.

[0032] In certain embodiments of the compound of formula (I), E is an optionally substituted aryl, preferably an optionally substituted phenyl.

[0033] In certain embodiments of the compound of formula (I), E is selected from one of the following by the removal of a hydrogen atom: [ka] (Each of these elements can be substituted as desired in the formula.)

[0034] In certain embodiments of the compound of formula (I), E is selected from the following groups: [ka] .

[0035] In certain embodiments of the compound of formula (I), E is selected from the following groups: [ka] .

[0036] In one embodiment of the present invention, the compound of formula (I) is represented by formula (Ia) or formula (Ib), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, R1, R2, Z, W, G, n, Y, E, R3, and R4 are as defined above.)

[0037] In a preferred embodiment, the compound of formula (I) has the stereochemistry shown in formula (Ib).

[0038] In one embodiment of the present invention, the compound of formula (I) is represented by formula (II), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, R1, R2, W, G, n, E, R3, and R4 are as defined above.)

[0039] In one embodiment of the present invention, the compound of formula (I) is represented by formula (III), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, W, G, n, Y, E, R3, and R4 are as defined above.)

[0040] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IV-1) to (IV-2), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, W, G, Y, E, R3, and R4 are as defined above.)

[0041] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (V-1) to (V-4), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, W, G, E, R 14 (R3 and R4 are as defined above).

[0042] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VI-1) to (VI-4), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] , (In the formula, A, W, G, E, R 14 (R3 and R4 are as previously defined). Preferably, W is an optionally substituted methyl group, and more preferably, W is -CH3 or -CF3.

[0043] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VII-1) to (VII-12), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] [ka] (In the formula, A, W, E, R 11 , R 12 , R 13 , R 14 (R3 and R4 are as previously defined). Preferably, W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3, and more preferably, W is -CH3 or -CF3.

[0044] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VIII-1) to (VIII-12), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, A, W, E, R 11 , R 12 , R 13 , R 14 (R3 and R4 are as previously defined). Preferably, W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3, and more preferably, W is -CH3 or -CF3.

[0045] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IX-1) to (IX-4), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [Chemical formula] (In the formula, each R 21 is independently optionally substituted methyl, halo, -CN, -OR 11 or -NR 11 R 12 and m is 0, 1, 2, 3, 4 or 5, and A, W, G, R 11 R 12 R 14 R3 and R4 are as defined above). Preferably, each R 21 is independently halo or optionally substituted methyl, and m is 1 or 2. More preferably, each R 21 is independently -F, -Cl, -CN, -CF3, -CH2F or -CHF2, and m is 1 or 2.

[0046] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (X-1) to (X-4), or a pharmaceutically acceptable salt, ester or prodrug thereof: [Chemical formula] , (In the formula, R 21 , m, A, W, G, R 14 R3, and R4 are as defined above). Preferably, each R 21 is independently halo or optionally substituted methyl, and m is 1 or 2.

[0047] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XII-1) to (XII-12), or a pharmaceutically acceptable salt, ester or prodrug thereof: [Chemical formula] (In the formula, m' is 0, 1, or 2). A, R3, R4, R 21 R 11 R 12 , and R 13is as defined above. Preferably, m' is 2.

[0048] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulae (XIII-1) to (XIII-12), or a pharmaceutically acceptable salt, ester or prodrug thereof:

Chemical formula

Chemical formula

[0049] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulae (XIV-1) to (XIV-6), or a pharmaceutically acceptable salt, ester or prodrug thereof:

Chemical formula

[0050] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XV-1) to (XV-6), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, W, m', R3, R4, R 21 , R 22 , R 11 and R 12 (This is as defined above). Preferably, R 21 R is a halogen, R3 is -OH, R4 is -CH3, -CF3, or cyclopropyl, and W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3. In certain embodiments, two adjacent R 22 The groups, together with the carbon atoms to which they are bonded, form a 4- to 12-membered carbocyclic or heterocyclic ring, which is condensed with phenyl or quinolinyl.

[0051] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XVI-1) to (XVI-12), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] [ka] (In the formula, R 23 is hydrogen; halo; -CN; -NO2, -OR 11 ;-NR 11 R 12 ;-NR 11 C(O)R 12 ;-NR 11 S(O)2R 12 ;-S(O)2R 12 ;-S(O)2NR 11 R 12 , -NR 11 C(O)NR 11 R 12 ;-C(O)R 11 , -C(O)OR 11 ;-C(O)NR 11 R 12 ;Optionally substituted -C1~C6 alkyl;Optionally substituted -C3~C8-cycloalkyl;Optionally substituted 3-membered to 8-membered heterocyclic;Optionally substituted aryl;Or optionally substituted heteroaryl, where W, R 21 , R 22 , m', R3, R4, R 11 and R 12 (This is as defined above). Preferably, R 21 R3 is a halogen, R4 is -OH, R4 is -CH3, -CF3, or cyclopropyl, and W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3.

[0052] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XVII-1) to (XVII-12), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (wherein, W, R 21 , R 22 , R 23 , m ’ , R3, R4, R 11 , and R 12 are as defined above). Preferably, R 21 is halogen, R3 is -OH, R4 is -CH3, -CF3, or cyclopropyl, and W is cyclopropyl, ethyl, -CH3, -CH2F, -CHF2, or -CF3.

[0053] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XVIII-1) to (XVIII-14), or a pharmaceutically acceptable salt, ester or prodrug thereof:

Chemical formula

Chemical formula

[0054] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XIX-1) to (XIX-14), or a pharmaceutically acceptable salt, ester or prodrug thereof:

Chemical formula

Chemical formula

[0055] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XX-1) to (XX-20), or a pharmaceutically acceptable salt, ester or prodrug thereof: [ka] [ka] (In the formula, each R 31 R is independently an optionally substituted -C1~C3 alkyl or halo, 32 These are, independently, halo, -OR 11 , -NR 11 R 12 , -NR 11 C(O)R 12 -C(O)NR 11 R 12 , -C(O)R 11 , optionally substituted -C1~C6 alkyl, or optionally substituted -C3~C8-cycloalkyl, R 11 , R 12 and R 14 (This is as defined above). Preferably, R 31 It is a halo, R 32 R is a halo, -NH2, optionally substituted methyl, optionally substituted cyclopropyl, 14 It is an optionally substituted cyclopropyl.

[0056] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XXI-1) to (XXI-20), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] [ka] (In the formula, R 31 , R 32 , R 11 , R 12 , and R 14 (This is as defined above). Preferably, R 31 It is a halo, R 32 R is a halo, -NH2, optionally substituted methyl, optionally substituted cyclopropyl, 14It is an optionally substituted cyclopropyl.

[0057] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XXII-1) to (XXII-14), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] [ka] (In the formula, R 24 is hydrogen or R 22 And W, R4, R 14 , R 21 , and R 22 (As previously defined). Preferably, R4 is an optionally substituted -C3-C6 cycloalkyl. More preferably, R4 is an optionally substituted cyclopropyl or optionally substituted cyclobutyl.

[0058] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XXIII-1) to (XXIII-14), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] [ka] (In the formula, W, R4, R 14 , R 21 , R 22 , and R 24 (As previously defined). Preferably, R4 is an optionally substituted -C3-C6 cycloalkyl. More preferably, R4 is an optionally substituted cyclopropyl or optionally substituted cyclobutyl, and W is an optionally substituted methyl.

[0059] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXIV), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, A, E, R3, and R4 are as previously defined).

[0060] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXIV), or is a pharmaceutically acceptable salt, ester, or prodrug thereof, where R3 is -OH and R4 is optionally substituted methyl or optionally substituted -C3-C6 cycloalkyl. A is [ka] Selected from, E is [ka] Select from, Alternatively, E is [ka] Selected from.

[0061] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXV), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, each R 41 , R 42 , R 43 , R 44 , and R 45 Each of these is independently selected from hydrogen, halogen, optionally substituted methyl, optionally substituted methoxyl, or -CN). R3, R4, R 14 , R 24This is as defined above. Preferably, R3 is -OH, and R4 is optionally substituted methyl or optionally substituted -C3~C6 cycloalkyl, R 14 R is an optionally substituted -C3~C6 cycloalkyl or optionally substituted methyl, 24 is optionally substituted with a methoxyl or halogen.

[0062] In one embodiment of the present invention, R 42 and R 43 Compounds of formula (XXV), or pharmaceutically acceptable salts, esters, or prodrugs thereof, which, together with the carbon atoms to which they are bonded, form condensed 4- to 7-membered carbocyclic or heterocyclic rings.

[0063] In one embodiment of the present invention, R 41 and R 42 Compounds of formula (XXV), or pharmaceutically acceptable salts, esters, or prodrugs thereof, which, together with the carbon atoms to which they are bonded, form condensed 4- to 7-membered carbocyclic or heterocyclic rings.

[0064] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XXVI-1) to (XXVI-3), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, R3, R 14 , R 24 , R 41 , R 42 , R 43 , R 44 , and R 45 (This is as defined earlier.)

[0065] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXVII), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, R 14 , R 24 , R 41 , R 42 , R 43 , R 44 , and R 45 (This is as defined earlier.)

[0066] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXVIII), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, R 14 , R 24 , R 41 , R 42 , R 43 , R 44 , and R 45 (This is as defined earlier.)

[0067] In one embodiment of the present invention, R 42 and R 43 Compounds of formula (XXVII) or formula (XXVIII), or pharmaceutically acceptable salts, esters, or prodrugs thereof, which, together with the carbon atoms to which they are bonded, form a condensed 4- to 7-membered carbocyclic ring or a heterocyclic ring.

[0068] In one embodiment of the present invention, R 43 and R 44 Compounds of formula (XXVII) or formula (XXVIII), or pharmaceutically acceptable salts, esters, or prodrugs thereof, which, together with the carbon atoms to which they are bonded, form a condensed 4- to 7-membered carbocyclic ring or a heterocyclic ring.

[0069] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (XXIV), (XXV), (XXVI-1) to (XXVI-3), (XXVII), or (XXVIII), or is a pharmaceutically acceptable salt, ester, or prodrug, where R 14R is an optionally substituted -C3~C6 cycloalkyl or optionally substituted methyl, 24 is a methoxyl or halogen which may be substituted, and each R 41 , R 42 , R 43 , R 44 and R 45 Each of these is independently selected from hydrogen, -F, -Cl, -CH3, -CF3, -OCH3, or -OCF3.

[0070] In one embodiment of the present invention, the compound of formula (I) is represented by formula (XXIV), formula (XXV), formula (XXVI-1) to (XXVI-3), formula (XXVII), or formula (XXVIII), or is a pharmaceutically acceptable salt, ester, or prodrug, where R 14 is -CHF2, [ka] or The file is JPEG0007871278000050.jpg819, and R 24 is -F, -Cl, or -OCH3, [ka] teeth, [ka] is or [ka] teeth, [ka] Selected from.

[0071] In one embodiment of the present invention, the compound of formula (I) is represented by formulas (XXIX-1) to (XXIX-3), or is a pharmaceutically acceptable salt, ester, or prodrug thereof: [ka] (In the formula, R 46 R is a substituted methyl or optionally substituted cyclopropyl, 47 is hydrogen, Cl, F, or an optionally substituted methoxyl, R 41 , R 42 , R 43 , R 44 and R 45 (This is as previously defined). In certain embodiments of the compounds of formulas (XXIX-1), (XXIX-2), and (XXIX-3), R 46 These are cyclopropyl, 1-fluorocyclopropyl, or difluoromethyl.

[0072] In one embodiment of the present invention, R 42 and R 43 However, together with the carbon atoms to which they are bonded, they form a condensed 4- to 7-membered carbocyclic ring or a heterocyclic ring, one of the compounds of formulas (XXIX-1) to (XXIX-3), or a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0073] In one embodiment of the present invention, R 43 and R 44 However, together with the carbon atoms to which they are bonded, they form a condensed 4- to 7-membered carbocyclic ring or a heterocyclic ring, one of the compounds of formulas (XXIX-1) to (XXIX-3), or a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0074] It will be understood that the description of the present invention herein should be interpreted in accordance with the laws and principles of chemical bonding. In some cases, it may be necessary to remove a hydrogen atom in order to accommodate a substituent at any given position.

[0075] The definition of any substituent or variable (e.g., R1, R2, etc.) at a specific location within a molecule is intended to be independent of its definition elsewhere within that molecule.

[0076] It will be further understood that the compounds of the present invention may contain one or more chiral carbon atoms and may exist in racemic, diastereoisomer, and optically active forms. It will still be understood that certain compounds of the present invention may exist in different tautomeral forms. It is intended that all tautomers are within the scope of the present invention.

[0077] In certain embodiments, the present invention provides a method for the prevention or treatment of RSV activity and for treating RSV infection in subjects requiring such treatment. This method involves administering a therapeutically effective amount of a compound of formula (I) to a subject.

[0078] The present invention also provides the use of a compound of formula (I) for preparing a pharmaceutical product for the prevention or treatment of RSV.

[0079] Therefore, in one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is combined with a steroidal anti-inflammatory compound, such as budesonide or fluticasone. In a preferred embodiment, the steroid is administered in a low dose to minimize immunosuppressive effects. In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is combined with a nonsteroidal anti-inflammatory compound, such as a leukotriene antagonist such as Singulair (Merck) or Accolate (Astra Zeneca), a phosphodiesterase 4 inhibitor such as roflumilast (Altana), a TNF-alpha inhibitor such as Enbrel (Amgen), Remicade (Centocor), Humira (Abbott) or CDP870 (Celltech), or an NSAID. In a further embodiment, the compound of formula (I) is combined with an interleukin 8 inhibitor or an interleukin 9 inhibitor. Accordingly, the present invention also relates to a product comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-inflammatory compound for simultaneous, separate, or sequential use in the treatment of RSV.

[0080] The present invention also relates to combinations of the compound of formula (I) or a pharmaceutically acceptable salt thereof with an anti-influenza compound, and to the use of such combinations in the treatment of simultaneous RSV and influenza infections. Accordingly, the present invention also relates to products containing the compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-influenza compound for simultaneous, separate, or sequential use in the treatment of simultaneous RSV and influenza infections. The compounds of the present invention can be administered in various dosage forms. Accordingly, they can be administered orally, for example, as tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules. The compounds of the present invention can also be administered parenterally, subcutaneously, intravenously, intramuscularly, intrasternally, percutaneously, or by infusion techniques. The compounds can also be administered as suppositories.

[0081] In one embodiment, the compounds of the present invention are administered by intranasal or intrabronchial administration. The present invention also provides an inhaler or nebulizer containing a pharmaceutical product comprising (a) a derivative of formula (I) as defined above or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier or diluent.

[0082] The present invention also provides a pharmaceutical composition containing such a benzodiazepine derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0083] The compounds of the present invention are typically formulated for administration with a pharmaceutically acceptable carrier or diluent. For example, a solid oral form may contain, together with the active compound, a diluent such as lactose, dextrose, saccharose, cellulose, corn starch, or potato starch; a lubricant such as silica, talc, stearic acid, magnesium stearate, or calcium stearate, and / or polyethylene glycol; a binder such as starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, or polyvinylpyrrolidone; a disaggregating agent such as starch, alginic acid, alginate, or sodium starch glycolate; a foaming mixture; a dye; a sweetener; a wetting agent such as lecithin, polysorbate, or lauryl sulfate; and generally, non-toxic and pharmacokinetically inert substances used in pharmaceutical formulations. Such pharmaceutical preparations may be produced by known methods, for example, by mixing, granulation, tableting, sugar coating, or film coating processes.

[0084] Liquid dispersions for oral administration may be syrups, emulsions, and suspensions. Syrups may contain, for example, saccharose or saccharose containing glycerin and / or mannitol and / or sorbitol as a carrier.

[0085] The suspensions and emulsions may contain, for example, natural rubber, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol as a carrier. Suspensions or solutions for intramuscular injection may contain, along with the active compound, a pharmaceutically acceptable carrier, such as sterile water, olive oil, ethyl oleate, glycol, such as propylene glycol, and, if necessary, an appropriate amount of lidocaine hydrochloride.

[0086] The solution for injection or infusion may contain, for example, sterile water as a carrier, or preferably in the form of sterile aqueous isotonic saline.

[0087] The present invention also relates to novel compounds as defined above, or pharmaceutically acceptable salts thereof, for use in methods of treating the body of a human or animal. The present invention also relates to pharmaceutical compositions comprising novel compounds as defined above and pharmaceutically acceptable diluents or carriers. Preferably, the pharmaceutical composition comprises a pharmaceutically acceptable salt of a novel compound as defined above. A pharmaceutically acceptable salt is as defined above. The novel compounds of the present invention are typically administered in the manner defined above, and the compounds are typically formulated for administration in the manner defined above.

[0088] Preferably, the pharmaceutical composition contains optically active isomers of the novel compound of the present invention. Therefore, for example, a preferred novel compound of the present invention containing only one chiral center includes a substantially pure form of the R enantiomer, a substantially pure form of the S enantiomer, and an enantiomer mixture containing an excess of the R enantiomer or excess of the S enantiomer. It is particularly preferable that the pharmaceutical composition contains the compound of the present invention, which is a substantially pure optical isomer. To avoid misunderstanding, the novel compound of the present invention may be used in solvate form as needed.

[0089] Further embodiments of the present invention are processes for producing any of the compounds described herein using any of the synthetic means described herein.

[0090] definition The following lists the definitions of various terms used to describe the present invention. These definitions apply to the terms used throughout this specification and the claims, individually or as part of a larger group, unless specifically limited in particular cases.

[0091] As used herein, the term "aryl" refers to, but is not limited to, monocyclic, bicyclic, or polycyclic carbocyclic ring systems containing at least one aromatic ring, including phenyl, naphthyl, tetrahydronaphthyl, indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system containing at least one aromatic ring. A polycyclic aryl may include fused rings, covalently bonded rings, or combinations thereof.

[0092] As used herein, the term “heteroaryl” refers to a monocyclic, bicyclic, or polycyclic aromatic radical having one or more ring atoms selected from S, O, and N, with the remaining ring atoms being carbon, and any N or S contained within the ring being optionally oxidized. Heteroaryls include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, and quinoxalinyl. Polycyclic heteroaryls may include fused rings, covalent rings, or combinations thereof.

[0093] According to the present invention, the aromatic group may be substituted or unsubstituted.

[0094] The terms "bicyclic aryl" or "bicyclic heteroaryl" refer to a ring system consisting of two rings, at least one of which is aromatic, and the two rings may be fused or covalently bonded.

[0095] As used herein, the term "alkyl" refers to saturated, linear or branched hydrocarbon radicals. Examples include "C1-C3 alkyl," "C1-C6 alkyl," and "C1-C 10"Alkyl," "C2-C4 alkyl," or "C3-C6 alkyl" refers to alkyl groups containing 1-3, 1-6, 1-10, 2-4, and 3-6 carbon atoms, respectively. Examples of C1-C8 alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, and octyl groups.

[0096] As used herein, the term “alkenyl” refers to a linear or branched hydrocarbon radical having at least one carbon-carbon double bond obtained by the removal of a single hydrogen atom. 10 "Alkenyl," "C2-C8 alkenyl," "C2-C4 alkenyl," or "C3-C6 alkenyl" refers to an alkenyl group containing 2-10, 2-8, 2-4, or 3-6 carbon atoms, respectively. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, and octenyl.

[0097] As used herein, the term "alkynyl" refers to a linear or branched hydrocarbon radical having at least one carbon-carbon triple bond obtained by the removal of a single hydrogen atom. 10 "Alkynyl," "C2-C8 alkynyl," "C2-C4 alkynyl," or "C3-C6 alkynyl" refers to an alkynyl group containing 2-10, 2-8, 2-4, or 3-6 carbon atoms, respectively. Typical alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, and octinyl.

[0098] As used herein, the term "cycloalkyl" refers to monocyclic or polycyclic saturated carbocyclic rings, or bicyclic or tricyclic group condensations, crosslinks, or spiro systems, where the carbon atoms may be oxosubstituted or optionally substituted with extracyclic olefins, imines, or oxime double bonds. Preferred cycloalkyl groups include C3-C 12Examples include cycloalkyl, C3-C6 cycloalkyl, C3-C8 cycloalkyl, and C4-C7 cycloalkyl. 12 Examples of cycloalkyl compounds include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, and spiro[4.4]nonanyl.

[0099] As used herein, the term "cycloalkenyl" refers to a monocyclic or polycyclic carbocyclic ring having at least one carbon-carbon double bond, or a bicyclic or tricyclic group condensation, bridge, or spiro system, wherein the carbon atoms may be oxosubstituted or optionally substituted with extracyclic olefins, imines, or oxime double bonds. Preferred cycloalkenyl groups include C3-C 12 Examples include cycloalkenyl groups, C3-C8 cycloalkenyl groups, or C5-C7 cycloalkenyl groups. 12 Examples of cycloalkenyls include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hepta-2-enyl, bicyclo[3.1.0]hexa-2-enyl, spiro[2.5]octa-4-enyl, spiro[4.4]nona-1-enyl, and bicyclo[4.2.1]nona-3-en-9-yl.

[0100] As used herein, the term "arylalkyl" refers to a functional group in which an alkylene chain is bonded to an aryl group, such as -CH2CH2-phenyl. The term "substituted arylalkyl" refers to an arylalkyl functional group in which an aryl group is substituted. Similarly, the term "heteroarylalkyl" refers to a functional group in which an alkylene chain is bonded to a heteroaryl group. The term "substituted heteroarylalkyl" refers to a heteroarylalkyl functional group in which a heteroaryl group is substituted.

[0101] As used herein, the term “alkoxy,” whether used alone or in combination with other terms, means an alkyl group having a specified number of carbon atoms bonded to the rest of the molecule via an oxygen atom, such as methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy), and their higher homologs and isomers, unless otherwise specified. Preferred alkoxys are (C1-C3)alkoxys.

[0102] It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, and cycloalkenyl moieties described herein may also be aliphatic or alicyclic groups.

[0103] An "aliphatic" group is a non-aromatic moiety composed of any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen, or other atoms, and may contain one or more unsaturated units, such as double and / or triple bonds. Examples of aliphatic groups include functional groups such as alkyl, alkenyl, alkynyl, O, OH, NH, NH2, C(O), S(O)2, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH2, S(O)2NH, S(O)2NH2, NHC(O)NH2, NHC(O)C(O)NH, NHS(O)2NH, NHS(O)2NH2, C(O)NHS(O)2, C(O)NHS(O)2NH, or C(O)NHS(O)2NH2, groups containing one or more functional groups, non-aromatic hydrocarbons (optionally substituted), and non-aromatic hydrocarbons (optionally substituted) in which one or more carbon atoms are replaced by a functional group. The carbon atoms of aliphatic groups may be oxo-substituted. The aliphatic group may be linear, branched, cyclic, or a combination thereof, and preferably contains about 1 to about 24 carbon atoms, more typically about 1 to about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, as used herein, the aliphatic group explicitly includes, for example, alkoxyalkyl groups, polyalkoxyalkyl groups, such as polyalkylene glycols, polyamines, and polyimines. The aliphatic group may be optionally substituted.

[0104] The term "carbocyclic ring" or "carbocyclic formula" refers to a saturated, partially unsaturated, or aromatic cyclic group in which each atom in the ring is carbon. Examples of carbocyclic rings include cycloalkyl groups, cycloalkenyl groups, and aryl groups.

[0105] The terms “heterocyclic” or “heterocycloalkyl” can be used interchangeably and refer to non-aromatic rings or bicyclic or tricyclic group condensations, bridges or spiro systems, where (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen; (ii) each ring system may be saturated or unsaturated; (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized; (iv) the nitrogen heteroatom may optionally be quaternized; (v) any of the above rings may be condensed to an aromatic ring; and (vi) the remaining ring atoms may optionally be oxosubstituted or optionally optionally substituted with an extracyclic olefin, imine or oxime double bond. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinozalinyl, pyridadinyl, 2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepant-4-yl, and tetrahydrofuryl. Such heterocyclic groups may be further substituted. The heteroaryl or heterocyclic group may be a C bond or an N bond (if possible).

[0106] It is understood that any alkyl, alkenyl, alkynyl, alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphatic moieties, etc., described herein may be divalent or polyvalent when used as a bond to link two or more groups or substituents that may be present on the same or different atoms. Those skilled in the art can readily determine the valency of any such group from the context in which it arises.

[0107] The term "substituted" refers to one, two, or three or more hydrogen atoms, but is not limited to -F, -Cl, -Br, -I, -OH, C1~C 12 -alkyl;C2~C 12 - Alkenil, C2~C 12 -Alkinyl, -C3~C 12 -Cycloalkyl, protected hydroxy, -NO2, -N3, -CN, -NH2, protected amino, oxo, thio, -NH-C1~C 12 -alkyl, -NH-C2~C8-alkenyl, -NH-C2~C8-alkynyl, -NH-C3~C 12 -Cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, -O-C1~C 12 -alkyl, -O-C2~C8-alkenyl, -O-C2~C8-alkynyl, -O-C3~C 12 -Cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C1~C 12 -alkyl, -C(O)-C2~C8-alkenyl, -C(O)-C2~C8-alkynyl, -C(O)-C3~C 12 -Cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C1~C 12 -alkyl, -CONH-C2~C8-alkenyl, -CONH-C2~C8-alkynyl, -CONH-C3~C 12 -Cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C1~C 12 -alkyl, -OCO2-C2~C8-alkenyl, -OCO2-C2~C8-alkynyl, -OCO2-C3~C 12 -Cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocycloalkyl, -CO2-C1~C 12Alkyl, -CO2-C2~C8 alkenyl, -CO2-C2~C8 alkynyl, CO2-C3~C 12 -Cycloalkyl, -CO2-aryl, CO2-heteroaryl, CO2-heterocycloalkyl, -OCONH2, -OCONH-C1~C 12 -alkyl, -OCONH-C2~C8-alkenyl, -OCONH-C2~C8-alkynyl, -OCONH-C3~C 12 -Cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocycloalkyl, -NHC(O)H, -NHC(O)-C1~C 12 -alkyl, -NHC(O)-C2~C8-alkenyl, -NHC(O)-C2~C8-alkynyl, -NHC(O)-C3~C 12 -Cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclo-alkyl, -NHCO2-C1~C 12 -alkyl, -NHCO2-C2~C8-alkenyl, -NHCO2-C2~C8-alkynyl, -NHCO2-C3~C 12 -Cycloalkyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C1~C 12 -alkyl, -NHC(O)NH-C2~C8-alkenyl, -NHC(O)NH-C2~C8-alkynyl, -NHC(O)NH-C3~C 12 -Cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH2, -NHC(S)NH-C1~C 12 -alkyl, -NHC(S)NH-C2~C8-alkenyl, -NHC(S)NH-C2~C8-alkynyl, -NHC(S)NH-C3~C 12 -Cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH)NH-C1~C 12-alkyl, -NHC(NH)NH-C2~C8-alkenyl, -NHC(NH)NH-C2~C8-alkynyl, -NHC(NH)NH-C3~C 12 -Cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C1~C 12 -alkyl, -NHC(NH)-C2~C8-alkenyl, -NHC(NH)-C2~C8-alkynyl, -NHC(NH)-C3~C 12 -Cycloalkyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C1~C 12 -alkyl, -C(NH)NH-C2~C8-alkenyl, -C(NH)NH-C2~C8-alkynyl, -C(NH)NH-C3~C 12 -Cycloalkyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)-C1~C 12 -alkyl, -S(O)-C2~C8-alkenyl, -S(O)-C2~C8-alkynyl, -S(O)-C3~C 12 -Cycloalkyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocycloalkyl, -SO2NH2, -SO2NH-C1~C 12 -alkyl, -SO2NH-C2~C8-alkenyl, -SO2NH-C2~C8-alkynyl, -SO2NH-C3~C 12 -Cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2-C1~C 12 -alkyl, -NHSO2-C2~C8-alkenyl, -NHSO2-C2~C8-alkynyl, -NHSO2-C3~C 12 -Cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, -CH2SO2CH3, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C3~C 12-Cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -S-C1~C 12 -alkyl, -S-C2~C8-alkenyl, -S-C2~C8-alkynyl, -S-C3~C 12 This refers to substitution by independently replacing with substituents including -cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl, or methylthiomethyl. In certain embodiments, substituents are independently selected from halo, preferably Cl and F; C1-C4-alkyl, preferably methyl and ethyl; halo-C1-C4-alkyl, e.g., fluoromethyl, difluoromethyl, and trifluoromethyl; C2-C4-alkenyl; halo-C2-C4-alkenyl; C3-C6-cycloalkyl, e.g., cyclopropyl; C1-C4-alkoxy, e.g., methoxy and ethoxy; halo-C1-C4-alkoxy, e.g., fluoromethoxy, difluoromethoxy, and trifluoromethoxy; -CN; -OH; NH2; C1-C4-alkylamino; di(C1-C4-alkyl)amino; and NO2. It is understood that aryl, heteroaryl, alkyl, etc., may be further substituted. In some cases, each substituent in the substitution is further optionally substituted with one or more groups, if possible, each group independently selected from C1-C4-alkyl-CF3, -OCH3, -OCF3, -F, -Cl, -Br, -I, -OH, -NO2, -CN, and -NH2.

[0108] In certain embodiments, the substituted alkyl, alkenyl, or alkoxy group is substituted with one or more halogen atoms, preferably fluorine or chlorine atoms. Examples of such substituted alkyl groups include fluoromethyl, difluoromethyl, and trifluoromethyl. Examples of such substituted alkoxy groups include fluoromethoxy, difluoromethoxy, and trifluoromethoxy.

[0109] As used herein, the terms "halo" or "halogen" refer to a fluorine, chlorine, bromine, or iodine atom, either alone or as part of another substituent.

[0110] As used herein, the term “optionally substituted” means that the group referred to may be substituted or unsubstituted. In one embodiment, the group referred to is optionally substituted with zero substituents, i.e., the group referred to is unsubstituted. In another embodiment, the group referred to is optionally substituted with one or more additional groups selected individually and independently from the groups described herein.

[0111] The term "hydrogen" includes hydrogen and deuterium. Furthermore, the enumeration of atoms includes other isotopes of that atom, insofar as the resulting compound is pharmaceutically acceptable.

[0112] In certain embodiments, compounds of each formula herein are defined as including isotope-labeled compounds. An “isotope-labeled compound” is a compound in which at least one atomic position is enriched to a level significantly greater than the natural abundance of a particular isotope of a given element. For example, one or more hydrogen atomic positions in a compound can be enriched with deuterium to a level significantly greater than the natural abundance of deuterium, for example, to at least 1%, preferably at least 20%, or at least 50%. Such deuterated compounds may be metabolized more slowly than their non-deuterated analogs, and therefore may exhibit a longer half-life when administered to a subject. Such compounds can be synthesized using methods known in the art, for example, by using deuterated starting materials. Unless otherwise stated, isotope-labeled compounds may be pharmaceutically acceptable.

[0113] As used herein, the term "hydroxyl-activating group" refers to an unstable chemical moiety known in the art to activate a hydroxyl group so that it is eliminated during synthetic procedures such as substitution or elimination reactions. Examples of hydroxyl-activating groups include, but are not limited to, mesylates, tosylates, triflates, p-nitrobenzoates, and phosphonates.

[0114] As used herein, the term "activated hydroxyl" refers to a hydroxyl group activated with one of the hydroxyl activating groups defined above, including, for example, mesylate, tosylate, triflate, p-nitrobenzoate, and phosphonate groups.

[0115] As used herein, the term “hydroxy protecting group” refers to an unstable chemical moiety known in the art to protect a hydroxyl group from undesirable reactions during a synthetic procedure. After the synthetic procedure, the hydroxy protecting groups described herein can be selectively removed. Hydroxy protecting groups known in the art are generally described in T. Greene and P. G. M. W. Tuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenyl-methyl(trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

[0116] As used herein, the term "protected hydroxyl" refers to a hydroxyl group protected by a hydroxyl protecting group as defined above, including, for example, benzoyl, acetyl, trimethylsilyl, triethylsilyl, and methoxymethyl groups.

[0117] As used herein, the term “hydroxyprodrug group” refers to a promoiety group, which is known in the art to transiently alter the physicochemical, and therefore biological, properties of a parent drug by coating or masking a hydroxyl group. After the synthesis procedure, the hydroxyprodrug group described herein must be able to revert to a hydroxyl group in vivo. Hydroxyprodrug groups known in the art are generally described in Kenneth B. Sloan, Prodrugs, Topical and Ocular Drug Delivery, (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992), and in “Prodrugs of Alcohols and Phenols,” VJStella, et al., Springer and AAPSPress, 2007, pp. 31-99, Prodrugs Challenges and Rewards Part-2, (Biotechnology: Pharmaceutical Aspects).

[0118] As used herein, the term “amino protecting group” refers to an unstable chemical moiety known in the art to protect an amino group from undesirable reactions during a synthetic procedure. After the synthetic procedure, the amino protecting groups described herein can be selectively removed. Amino protecting groups known in the art are generally described in T. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl, and benzyloxycarbonyl.

[0119] As used herein, the term "protected amino" refers to an amino group protected by the amino protecting group defined above.

[0120] The term "leaving group" refers to a functional group or atom that can be substituted by another functional group or atom in substitution reactions such as nucleophilic substitution reactions. Examples of typical leaving groups include chloro, bromo, and iodine groups; sulfonic acid ester groups, such as mesylate, tosylate, brosylate, and nosylate; and acyloxy groups, such as acetoxy and trifluoroacetoxy.

[0121] As used herein, the term “aprotic solvent” refers to a solvent that is relatively inert to proton activity, i.e., does not act as a proton donor. Examples include, but are not limited to, hydrocarbons such as hexane and toluene, halogenated hydrocarbons such as methylene chloride, ethylene chloride, and chloroform, heterocyclic compounds such as tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether and bis-methoxymethyl ether. Such compounds are well known to those skilled in the art, and it will be apparent to those skilled in the art that, depending on factors such as reagent solubility, reagent reactivity, and preferred temperature range, individual solvents or mixtures thereof may be preferred for particular compounds and reaction conditions. Further discussion of aprotic solvents can be found in organic chemistry textbooks or specialized monographs, for example, in *Organic Solvents Physical Properties and Methods of Purification*, 4th ed., edited by John A. Riddick et al., Vol. II, in *Techniques of Chemistry Series*, John Wiley & Sons, NY, 1986.

[0122] As used herein, the term “protic solvent” refers to solvents that tend to provide protons, such as alcohols, e.g., methanol, ethanol, propanol, isopropanol, butanol, t-butanol, etc. Such solvents are well known to those skilled in the art, and it will be apparent to them that, depending on factors such as reagent solubility, reagent reactivity, and preferred temperature range, individual solvents or mixtures thereof may be preferred for certain compounds and reaction conditions. Further discussion of protic solvents can be found in organic chemistry textbooks or specialized monographs, for example, in *Organic Solvents: Physical Properties and Methods of Purification*, 4th ed. (edited by John A. Riddick et al.), Vol. II, in the *Techniques of Chemistry Series*, John Wiley & Sons, NY, 1986.

[0123] The combinations of substituents and variables envisioned in the present invention simply result in the formation of stable compounds. As used herein, the term “stable” means a compound that is stable enough to enable production and maintains the integrity of the compound over a sufficient period of time to serve the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0124] The synthesized compound can be separated from the reaction mixture and further purified by methods such as column chromatography, high-pressure liquid chromatography, or recrystallization. Further methods for synthesizing the compounds of the formulas herein will be obvious to those skilled in the art, as can be understood by them. Furthermore, the desired compounds can be obtained by carrying out various synthetic steps in an alternative order or sequence. Useful synthetic chemical transformations and protecting group methodologies (protection and deprotection) for synthesizing the compounds described herein are known in the art, for example, R. Larock, Comprehensive Organic Transformations, 2 ndThis includes the works described in Ed. Wiley-VCH (1999); T. Greene and P. G. M. Tuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions.

[0125] As used herein, the term “subject” refers to an animal. Preferably, the animal is a mammal. More preferably, the mammal is a human. The subject also refers to, for example, dogs, cats, horses, cattle, pigs, guinea pigs, fish, birds, etc.

[0126] The compounds of the present invention can be modified by adding appropriate functional groups to enhance their selective biological properties. Such modifications are known in the art and may include those that increase biopenetration into a given biological system (e.g., blood, lymphatic system, central nervous system), enhance oral availability, increase solubility to enable administration by injection, alter metabolism, and alter excretion rate.

[0127] The compounds described herein contain one or more chiral centers, thus giving rise to enantiomers, diastereomers, and other stereoisomers, which can be defined from the viewpoint of absolute stereochemistry as (R)- or (S)-, or with respect to amino acids as (D)- or (L)-. The present invention means that all such possible isomers are included as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors by the above procedure or by resolution of racemic mixtures. Resolution can be carried out by chromatography in the presence of a resolving agent, by repeated crystallization, or by some combination of these techniques known to those skilled in the art. Further details regarding resolution can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Where the compounds described herein contain olefinic double bonds, other unsaturated or other geometrically chiral centers, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers or cis and trans isomers. Similarly, all tautomers are intended to be included. Tautomers may be cyclic or acyclic. Any carbon-carbon double bond configurations appearing herein are selected for convenience only and are not intended to specify any particular configuration unless otherwise stated herein. Thus, any carbon-carbon double bond or carbon-heteroatom double bond optionally shown as trans herein may be cis, trans, or a mixture of any two in any proportion.

[0128] Certain compounds of the present invention may also exist in different stable conformations that may be separable. Torsional chirality resulting from restricted rotation around a chiral single bond, for example, due to steric hindrance or ring strain, may allow for the separation of different conformational isomers. The present invention includes each conformational isomer of these compounds and mixtures thereof.

[0129] As used herein, the term “pharmaceutically acceptable salt” refers to a salt that, within the bounds of sound medical judgment, is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, allergic reactions, etc., and that provides a reasonable benefit-risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). Salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reacting the free basic functional group with a suitable organic acid. Examples of pharmaceutically acceptable salts, but not limited to, include non-toxic acid addition salts of amino groups formed by using inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids, such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentane-propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobacillate. Examples of alkali metal salts include, but are not limited to, ions, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfons, 2-naphthalenesulfons, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectins, persulfates, 3-phenylpropionates, phosphates, pivalates, picrates, propions, stearates, succinates, sulfates, tartrates, thiocyans, p-toluenesulfons, undecanoates, and valersates. Typical alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium.Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium compounds, quaternary ammonium compounds, and amine cations formed using counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and arylsulfonates.

[0130] pharmaceutically acceptable salts can also be prepared by deprotonating the parent compound with a suitable base, thereby forming an anionic conjugate base of the parent compound. In such salts, the counterion is a cation. Suitable cations include ammonium and metal cations, such as alkali metal cations, such as Li. + na + , K + and Cs + , as well as alkaline earth metal cations, such as Mg 2+ and Ca 2+ These are some examples.

[0131] As used herein, the term “pharmaceutically acceptable ester” refers to an ester that hydrolyzes in vivo and includes those that readily decompose in the human body, leaving the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanes, alkenes, cycloalkanoates, and alkanedioates, where each alkyl or alkenyl moiety has preferably six or fewer carbon atoms. Examples of specific esters include, but are not limited to, esters of C1-C6 alkanes such as acetates, propions, butyrates, and pivalates.

[0132] In certain embodiments, the present invention provides pharmaceutically acceptable prodrugs of compounds disclosed herein. As used herein, the term “pharmaceutically acceptable prodrug” refers to a prodrug of a compound formed by the process of the present invention that is suitable for use in contact with human and lower animal tissues that have excessive toxicity, irritation, allergic reactions, etc., within the bounds of sound medical judgment, is commensurate with a reasonable benefit / risk ratio, and is effective for their intended use, as well as, if possible, a zwitterionic form of the compound of the present invention. As used herein, “prodrug” means a compound that is convertible in vivo by metabolic means (e.g., by hydrolysis) and gives any compound described by the formula of the present invention. Various forms of prodrugs are known in the art, for example, Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed.). "Design and Application of Prodrugs," Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8: 1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77: 285 et seq. (1988); Higuchi and Stella (eds.), Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975); and Bernard Testa & Joachim Mayer, "Hydrolysis In Drug And This is discussed in "Prodrug Metabolism: Chemistry, Biochemistry and Enzymology," John Wiley and Sons, Ltd. (2002).

[0133] Additional types of prodrugs are also included. For example, free carboxyl groups can be derivatized as amides or alkyl esters. Free hydroxyl groups can be derivatized using groups including, but not limited to, hemisuccinates, ethyl succinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyl, as outlined in Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxyl and amino groups are also included, as are carbonate prodrugs, sulfonate esters, and sulfate esters of hydroxyl groups. Derivatization of hydroxyl groups as (acyloxy)methyl and (acyloxy)ethyl ethers is also included, where the acyl group may be an alkyl ester and optionally substituted with groups including, but not limited to, ethers, amines, and carboxylic acid functional groups, or where the acyl group is one of the above amino acid esters. This type of prodrug is described in J.Med.Chem. 1996, 39, 10. Free amines can also be derivatized as amides, sulfonamides, or phosphoamides. All of these prodrug portions may incorporate groups including, but not limited to, ether, amine, and carboxylic acid functional groups. In certain embodiments, the compounds of the present invention may incorporate two or more groups that are metabolically removed in vivo to produce the active parent compound.

[0134] As used herein, the term “treatment” means reducing, decreasing, mitigating, eliminating, regulating, or improving a disease state or symptom, i.e., causing a regression of the disease state or symptom. Treatment may also include, for example, inhibiting an existing disease state or symptom, i.e., stopping its onset, and alleviating or improving an existing disease state or condition, i.e., causing its regression, if such a disease state or symptom may already exist.

[0135] As used herein, the term “prevention” means completely or almost completely stopping the occurrence of a disease condition or symptom in a patient or subject, especially when the patient or subject is susceptible to or at risk of developing such a disease condition or symptom.

[0136] Furthermore, the compounds of the present invention, such as salts of the compounds, can exist in hydrated or unhydrated (anhydrous) forms, or as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates and dihydrates. Non-limiting examples of solvates include ethanol solvate and acetone solvate.

[0137] A "solvate" refers to a solvation form containing a stoichiometric or non-stoichiometric amount of solvent. Some compounds tend to capture a certain molar ratio of solvent molecules in their crystalline solid state, and thus form solvates. When the solvent is water, the solvate formed is a hydrate; when the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by a combination of one or more molecules of water and one of the substances in which water retains its molecular state as H2O, and such combinations can form one or more hydrates.

[0138] As used herein, the term “analog” refers to a chemical compound that is structurally similar to another but has a slightly different composition (e.g., substitution of one atom by an atom of a different element, or substitution of one functional group by another functional group). Thus, an analog is a compound that is similar or equivalent to a reference compound in function and appearance.

[0139] The combinations of substituents and variables envisioned in the present invention simply result in the formation of stable compounds. As used herein, the term “stable” means a compound that is stable enough to enable production and maintains the integrity of the compound over a sufficient period of time to serve the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0140] The synthesized compound can be separated from the reaction mixture and further purified by methods such as column chromatography, high-pressure liquid chromatography, or recrystallization. Furthermore, the desired compound can be obtained by carrying out various synthesis steps in an alternative order or sequence. Moreover, the solvents, temperatures, reaction times, etc., described herein are for illustrative purposes only, and variations in reaction conditions can produce the desired crosslinked macrocyclic product of the present invention. Useful synthetic chemical transformations and protecting group methodologies (protection and deprotection) for synthesizing the compounds described herein include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); TW. Greene and PG. MWuts, Protective Groups in Organic Synthesis, 2nd ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

[0141] The compounds of the present invention may be modified by adding various functional groups via synthetic means described herein to enhance their selective biological properties. Such modifications may include increasing biopenetration into a given biological system (e.g., blood, lymphatic system, central nervous system), improving oral availability, increasing solubility to enable administration by injection, altering metabolism, and altering excretion rate.

[0142] Pharmaceutical composition The pharmaceutical composition of the present invention comprises a therapeutically effective amount of the compound of the present invention formulated with one or more pharmaceutically acceptable carriers. As used herein, the term “pharmaceutically acceptable carrier” means any kind of non-toxic inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch, potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository waxes; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil; glycols, e.g., propylene glycol; esters, e.g., ethyl oleate, ethyl laurate; agar; buffers, e.g., magnesium hydroxide, aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; and ethyl alcohol. Phosphate buffer, as well as other non-toxic compatible lubricants, e.g., sodium lauryl sulfate and magnesium stearate, and colorants, release agents, coating agents, sweeteners, flavoring agents, and fragrances, preservatives, and antioxidants may also be present in the composition at the discretion of the compounder. The pharmaceutical compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisionally, vaginally, intraperitoneally, topically (as in powder, ointment, or drops), orally or as a nasal spray.

[0143] The pharmaceutical compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir, preferably by oral or injectable administration. The pharmaceutical compositions of the present invention may contain any conventional non-toxic, pharmaceutically acceptable carrier, adjuvant, or vehicle. In some cases, the pH of the formulation may be adjusted with a pharmaceutically acceptable acid, base, or buffer to enhance the stability of the formulated compound or its delivery form. As used herein, the term parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-arterial, intra-sacral, intrasternal, intrathecal, intrafocal, and intracranial injection or infusion techniques.

[0144] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, as well as mixtures thereof. In addition to inert diluents, the oral composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0145] Preparations for injection, such as sterile aqueous or oily suspensions for injection, can be formulated according to known techniques using appropriate dispersants or wetting and suspending agents. Sterile preparations for injection may also be sterile solutions, suspensions, or emulsions in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, USP, and isotonic sodium chloride solutions. Furthermore, sterile fixatives have been conventionally used as solvents or suspension media. For this purpose, any non-irritating fixative containing synthetic monoglycerides or diglycerides can be used. Additionally, fatty acids such as oleic acid are used in preparations for injection.

[0146] Injectable formulations can be sterilized, for example, by filtration using a bacterial capture filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0147] To prolong the effects of a drug, it is often desirable to delay the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with low water solubility. The absorption rate of a drug depends on its dissolution rate, which may then depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is achieved by dissolving or suspending the drug in an oily vehicle. Depot formulations for injection are prepared by forming a microcapsule matrix of the drug in a biodegradable polymer such as polylactide-polyglycolide. The drug release rate can be controlled depending on the drug-to-polymer ratio and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydrous). Depot injection formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0148] The composition for rectal or vaginal administration is preferably a suppository, which can be prepared by mixing the compound of the present invention with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, or suppository wax, which is solid at ambient temperature but liquid at body temperature and therefore melts in the rectal or vaginal cavity to release the active compound.

[0149] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is provided with at least one inert, pharmaceutically acceptable excipient or carrier, e.g., sodium citrate or dicalcium phosphate and / or: a) fillers or bulking agents, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, e.g., carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) wetting agents, e.g., glycerol; d) disintegrants, e.g., agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarding agents. f) an absorption enhancer, such as paraffin; g) a wetting agent, such as cetyl alcohol and glycerol monostearate; h) an absorbent, such as kaolin and bentonite clay; and i) a lubricant, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include a buffer.

[0150] Similar types of solid compositions can also be used as fillers in soft and rigid gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0151] The active compound may also be in a microencapsulated form with one or more excipients, as described above. Solid dosage forms of tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coatings and shells, such as enteric coatings, controlled-release coatings, and other coatings well known in the pharmaceutical field. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also contain additional substances other than the inert diluent, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose, as is common practice. In the case of capsules, tablets, and pills, the dosage form may also contain buffers. These may optionally contain opacifiers and may be compositions that release the active ingredient alone or preferentially, in a delayed manner, in a specific part of the intestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0152] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed with a pharmaceutically acceptable carrier under sterile conditions and, if necessary, with any required preservatives or buffers. Ophthalmic formulations, ear drops, eye ointments, powders, and solutions are also considered to be within the scope of the present invention.

[0153] The ointments, pastes, creams, and gels may contain, in addition to the active compound of the present invention, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0154] The powders and sprays may contain, in addition to the compounds of the present invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures thereof. The sprays may further contain conventional spraying agents such as chlorofluorohydrocarbons.

[0155] Transdermal patches offer the additional advantage of controlling the delivery of compounds to the body. Such dosage forms can be prepared by dissolving or aliquoting the compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound across the skin. The rate can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0156] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly known to those skilled in the art. All publications, patents, published patent applications, and other references referenced herein are incorporated herein by reference in their entirety.

[0157] Abbreviation The abbreviations used in the following explanation of the scheme and examples are as follows: ACN: Acetonitrile; AD-mix-β:(9S)-(9''S)-9,9''-[1,4-phthalazinediylbis(oxy)]bis[10,11dihydro-6'-methoxycinconan]; Bn: benzyl; BOP: (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate; BzCl: Benzoyl chloride; mCPBA: Meta-chloroperbenzoic acid; Cbz: Benzyloxycarbonyl; CDI: Carbonyldiimidazole; DAST: Diethylaminosulfur trifluoride; DBU:1,8-diazabicycloundecene-7ene; DCE: Dichloroethane; DCM: Dichloromethane; Des Martinperiodinane:1,1,1-Tris(acetyloxy)-1,1-dihydro-1,2-benzoiodoxol-3-(1H)-one; DIAD: Diisopropyl azodicarboxylate; DIBAL-H: Diisobutylaluminum hydride; DMAP: N,N-dimethylaminopyridine; DME: 1,2-dimethoxyethane; DMF: N,N-dimethylformamide; DMSO: Dimethyl sulfoxide; DPPA: Diphenylphosphoryl azide or diphenylphosphoryl azide; dppf: 1,1'-bis(diphenylphosphin)ferrocene; EDCI or EDC: 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride; EA or æ: ethyl acetate; Ghosez reagent: 1-chloro-N,N,2-trimethyl-1-propenylamine; HATU:O(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; HCl: hydrochloric acid; Hünig base: diisopropylethylamine; PyBOP: (benzotriazole-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate; LDA: Lithium diisopropylamine; Pd-C: Palladium-carbon; PE: Petroleum ether; Ph: Phenyl; RT: Reverse transcription; RT-PCR: Reverse transcription polymerase chain reaction; TBME: tert-butyl methyl ether; TEA: Triethylamine; Tf2O: Trifluoromethanesulfonic anhydride; TFA: Trifluoroacetic acid; THF: Tetrahydrofuran; (TMS)2NH; Hexamethyldisilazane; TBS: tert-butyldimethylsilyl; TBDPS: tert-butyldiphenylsilyl; TMS: Trimethylsilyl; TPAP: Tetrapropylammonium perlutenate; TPP or PPh3: Triphenylphosphine; Ts or Tosyl: p-CH3C6H4SO2-; tBOC or Boc:tert-butyloxycarbonyl; and Xanthophos: 4,5-bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene.

[0158] Synthesis method The compounds and processes of the present invention will be better understood in connection with the following synthetic scheme (intended for illustrative purposes only and not to limit the scope of the invention) illustrating a method by which the compounds of the present invention can be prepared. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications, including but not limited to those relating to the chemical structure, substituents, derivatives and / or methods of the present invention, can be made without departing from the spirit of the invention and the scope of the appended claims.

[0159] Scheme 1 shows a method for preparing the compound of formula 11 from compounds 1 and 2, where n = 1, 2, or 3, P is a hydroxy protecting group, Ar is E, and E is as previously defined. Alkylation of hydroxypyridine 1 with a hydroxy epoxide using Mitsunobu reaction conditions yields epoxide 4. Alternatively, hydroxy epoxide is converted to 3 having a leaving group, such as tosyl and methanesulfonyl (but not limited to), and then alkylated in the presence of a base, such as K2CO3 and Cs2CO3 (but not limited to), to yield 4. Intramolecular epoxide ring-opening mediated by a base, such as LDA (but not limited to), produces compound 5. Protection of the hydroxy compound 5 with a suitable protecting group, such as TBDPS and TBS (but not limited to), yields compound 6. Trifluoromethyl ketone 7 is obtained by iodine-magnesium exchange of compound 6 followed by the addition of an ester, such as ethyl 2,2,2-trifluoroacetate (but not limited to). Compound 9 is obtained by cross-coupling trifluoromethyl ketone 7 with various metal coupling partners 8, such as boronic acids, boronic acid esters, organotin reagents, organozinc reagents, organomagnesium reagents, and organosilicon reagents, using a suitable catalyst such as Pd, Ni, or Cu. Compound 10 is obtained by nitromethane addition to compound 9 in the presence of bases such as K2CO3 and Cs2CO3, although this method is not limited to these. Reduction of the nitro group with reducing reagents such as zinc and acetic acid generates an important intermediate 11, although this method is not limited to these. Scheme 1 [ka]

[0160] As seen in Scheme 2, Ar1 is A, Ar is E, and R is R 11 n is 1, 2, or 3, and A, E, R 11The definition is as previously described. The important intermediate 11 is coupled with various carboxylic acids to obtain amide 14. Then, amide 14 is reacted with various electrophiles to produce various ethers, esters, and carbamates of formula 15. Amide 14 is also oxidized to aldehyde 16, and then various amines 17 are obtained by reductive amination. The hydroxyl group -CH2OH in amide 14 is converted to cyanomethyl 18 by activation and subsequent cyanation. Compound 14-1 is further converted to acetamide 19 in the presence of a catalyst, such as a Perkin catalyst, but is not limited to this. Scheme 2 [ka]

[0161] As seen in Scheme 3, Ar1 is A, Ar is E, and R is R 11 A, E, R 11 The definition is as previously described. Aldehyde 16 is converted to a benzyl-protected amine by reductive amination. Hydrolysis yields the free amine 20. Finally, substitution with various electrophiles yields the N-substituted compound 21. Scheme 3 [ka]

[0162] As shown in Scheme 4, Ar1 is A, Ar is E, R' is -C1~C6 alkyl, -C3~C6 cycloalkyl, aryl, or heteroaryl, n is 1, 2, or 3, and A and E are as previously defined. After oxidizing aldehyde 16 to acid 22, it is further converted to amide 23 and sulfonamide 24 using general methods such as, but not limited to, HATU and DIPEA. From there, diversification into various esters and amides is carried out. Scheme 4 [ka]

[0163] Scheme 5 illustrates another method for preparing the compound of formula 11, where Ar is E, P is a hydroxyl protecting group, n is 1, 2, or 3, and E is as previously defined. Ketone 9 is converted to the compound of formula 26 via olefination. Alternatively, 26 can be obtained from: 1) a boronic acid, boronic ester, organotin reagent, organozinc reagent, organomagnesium reagent, organosilicon reagent, etc., which is catalyzed with a suitable catalyst such as Pd, Ni, or Cu to obtain compound 25 from 6 via cross-coupling with a metallic bonding partner 6-1; 2) compound 25 is converted to compound 26 as previously described in Scheme 1. With 26 prepared, the compound of formula 27 is prepared by dihydroxylation and subsequent epoxide formation. Epoxide ring-opening of compound 27 having amine equivalents such as but not limited to NH4OH and NH3 provides the compound of formula 11. Scheme 5 [ka]

[0164] Scheme 6 illustrates another method for preparing the compound of formula 23, where Ar1 is A, Ar is E, R' is -C1~C6 alkyl, -C3~C6 cycloalkyl, aryl, or heteroaryl, n is 1, 2, or 3, and A and E are as previously defined. Amine 11 is protected with protecting groups such as, but not limited to, Boc and Cbz. After deprotection of the hydroxyl protecting group, acid 30 is obtained by subsequent oxidation. Compound 30 is coupled with various amines to obtain amide 31. After deprotection of the amine protecting group and subsequent amide formation, the compound of formula 23 is obtained. Scheme 6 [ka]

[0165] Scheme 7 shows an additional route for synthesizing the desired compound. The difference in this route is that it begins with oxidation and amide coupling to introduce amide 33 at the start of the synthesis. Sequential vinylization and arylation yield the biscoupling product 34. Asymmetric dihydroxylation followed by activation and substitution yields the amino alcohol precursor. Finally, amide coupling with each aryl acid produces the desired compound represented by compound 36. Scheme 7 [ka]

[0166] example The compounds and processes of the present invention are intended to be illustrative only and will be better understood in relation to the following examples, which are not intended to limit the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications, including but not limited to those relating to the chemical structure, substituents, derivatives, formulations and / or methods of the present invention, can be made without departing from the spirit of the invention and the scope of the appended claims.

[0167] Specific synthetic procedures useful for preparing the compounds of the present invention are disclosed in U.S. Patent Application No. 16 / 930,622, which are incorporated herein by reference in their entirety.

[0168] Example 1 [ka] Example 1 Process a [ka] A solution of 3-bromo-4-hydroxybenzoic acid (16 g, 69.25 mmol), Cs2CO3 (68 g, 207.75 mmol), KI (46 g, 277.00 mmol), and bromocyclopropane (21 g, 173.12 mmol) in NMP (30 mL) was stirred in a Parr reactor at 180 °C for 16 hours. The resulting solution was diluted with water and extracted with EtOAC. The combined organic matter was dried and concentrated. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product as a yellow solid (3 g, 22%). ESI-MS m / z: 257.05 [M+H] + (The methyl ester product was also isolated and used.)

[0169] Example 1 Process b [ka] The compound from step a (250 mg, 0.98 mmol), Pd(dppf)Cl2 (142 mg, 0.19 mmol), 2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (425 mg, 1.94 mmol), H2O (0.1 mL), and Cs2CO3 (950 mg, 2.91 mmol) in dioxane (3 mL) were stirred at 90°C for 2 hours under an N2 atmosphere. The resulting solution was purified by reverse-phase C18 column chromatography (0.1% FA in MeOH / H2O) to obtain the desired product as a white solid (180 mg, 68%). ESI-MS m / z: 270.15 [M+H] + .

[0170] Example 1 Process c [ka] In a 2-drum vial equipped with a stirring bar, amine (30 mg, 0.075 mmol) and acid (19.18 mg, 0.075 mmol) were added, and the material was dissolved in DMF (0.2 M). Hünig base (0.053 mL, 0.30 mmol) was added, and the vial was cooled to 0°C. HATU (43 mg, 0.113 mmol) was added, the reaction mixture was stirred for 10 minutes, warmed to room temperature, and monitored by LC-MS (1 hour). The reaction mixture was diluted with RINKAN and quenched with water. The aqueous phase was extracted with RINKAN and DCM / MeOH using a phase separator cartridge and concentrated. The material was purified by preparative HPLC (20-90%) in MeCN / water for 25 minutes, and the title compound was obtained as a white solid (23.6 mg, 48%). ESI-MS m / z: 651.25 [M+H] + .

[0171] Example 2 [ka] Example 2 Process a [ka] In a 50 mL round-bottom flask equipped with a stirring bar, methyl 8-hydroxyquinoline-6-carboxylate (1,500 g, 7.38 mmol) and potassium carbonate (2,040 g, 14.76 mmol) were added, and the solid was dissolved in DMF (0.5 M). Then, tert-butyl 2-bromoacetate (1,308 ml, 8.86 mmol) was added, and the reaction mixture was stirred at 40°C and monitored by LC-MS (2 hours). The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and quenched with water. The water was extracted with ethyl acetate, the combined organic layers were dried, filtered, and concentrated. The residue was analyzed by automated column chromatography (silica gel, hexane, ethyl acetate). f Purification by ESI-MS (=0.27) yielded a white solid (1.93g, 82%). ESI-MS m / z:262.0 [M+H] + .

[0172] Example 2 Step b [ka] A stirring bar was added to a 100 mL round-bottom flask containing step a (1.93 g, 6.08 mmol), and the solid was dissolved in DCM (0.5 M). The flask was cooled to 0°C, and TFA (4.69 ml, 60.8 mmol) was added. The reaction mixture was stirred for 10 minutes, warmed to room temperature, and monitored by LC-MS (after 3 hours, an additional 5.0 equivalents of TFA were added; total 5.5 hours). The mixture was quenched with water and diluted with DCM. The solid precipitated. It was further diluted with DCM and stirred vigorously for 10 minutes. The solid was recovered by filtration, washed multiple times with DCM, and dried under high vacuum to obtain a light brown, fluffy solid (2.21 g, 97%). ESI-MS m / z: 262.0 [M+H] + .

[0173] Example 2 Process c [ka] Step b (125 mg, 0.333 mmol) was added to a 40 mL vial equipped with a stirring bar. The solid was dissolved in DMF and cooled to 0°C. After adding DIPEA (407 μl, 2.332 mmol), 1-methylcyclopropane-1-amine hydrochloride (124 mg, 1.148 mmol) was added. Then, PyBOP (260 mg, 0.500 mmol) was added all at once, the reaction mixture was stirred for 10 minutes, warmed to room temperature, and monitored by LC-MS (1.5 hours). The reaction mixture was diluted with RINKAN and quenched with water. The aqueous layer was extracted with RINKAN using a phase separator cartridge, and the combined organic layers were concentrated. The residue was purified to the title compound (98 mg, 82%) by automated column chromatography (silica gel, dichloromethane, 0-20% methanol). ESI-MS m / z: 216.0 [M+H] + .

[0174] Example 2 Process d [ka] General notes on hydrolysis: In some cases, the reactants were heated to 45°C to dissolve the materials and accelerate hydrolysis. After hydrolysis, the product was isolated by precipitation. If no precipitate was present, the product was extracted or the aqueous solution was concentrated (the materials were dried and the crude product was used). Major MS of all these compounds. + m / z is CC cut. ESI-MS m / z: 202.0 [M+H] + .

[0175] A stirring bar was added to a 20 mL vial containing step c (9 mg, 0.312 mmol) from Example 2. The compound was dissolved in MeOH, THF, and water (0.2 M, 2:1:1). Lithium hydroxide hydrate (62 mg, 1.56 mmol) was added, and the reaction was stirred at room temperature and monitored by LC-MS. The stirring bar was removed, and the vial was cooled to 0°C. The reaction was acidified with 2 M HCl to a pH of approximately 4-5 (1 M NaOH was used if it was too acidic). The product was extracted three times with 10% MeOH / DCM using a phase separator and concentrated. It was dried under high vacuum to obtain the title compound (45 mg, 50%). ESI-MS m / z: 202.0 [M+H] + .

[0176] Example 2 Process e [ka] The following example was prepared using the same procedure as in step c of Example 1, with the corresponding acid and amine HCl salt (25 mg) coupling partners from step d. The residue was purified by Gilson preparative HPLC (20-90%, MeCN / water, 25 min) to obtain the title compound (8 mg, 20%) ESI-MS m / z: 682.2.

[0177] Intermediate 1 [ka] Intermediate 1 step a [ka] In a vial, 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (566 mg, 3.67 mmol), methyl 2-bromobenzo[d]thiazole-6-carboxylate (500 mg, 1.837 mmol), PdCl2(dppf)DCM (75 mg, 0.092 mmol), and Na2CO3 (302 mg, 2.85 mmol) were dissolved in dioxane (6.9 ml) and water (2.3 ml). The reaction mixture was heated at 100°C for 3 hours. The reaction mixture was cooled to room temperature and diluted with water. The aqueous layer was washed with ethyl acetate. The combined organic layers were dried over MgSO4 and concentrated. The crude reaction mixture was purified by silica gel chromatography eluting with 0-50% ethyl acetate / hexane to obtain the title compound (180 mg, 0.821 mmol, 45%).

[0178] Intermediate 1 Steps b and c [ka] In a vial, the compound from step a (120 mg, 0.547 mmol) was dissolved in MeOH (5.47 ml). Pd / C (5.82 mg, 0.055 mmol) was added, and the vial was purged with H2. The reaction mixture was stirred under H2 at room temperature for 1 hour. The reaction mixture was purged with N2, and filtered through celite. The crude reaction mixture was concentrated and purified by silica gel chromatography eluted with 0-50% Â / hexane to obtain the title compound (80 mg, 0.362 mmol, 66%).

[0179] Ester hydrolysis was carried out in the same manner as in step d of Example 2. The title compound was isolated by precipitation to obtain the desired product (50 mg, 0.241 mmol, 67%).

[0180] Intermediate 2 [ka] In a vial, cyclopropyltrifluoro-14-borane, potassium salt (326 mg, 2.205 mmol), methyl 2-bromobenzo[d]thiazole-6-carboxylate (200 mg, 0.735 mmol), palladium tetrakis (42.5 mg, 0.037 mmol), and potassium phosphate (468 mg, 2.205 mmol) were dissolved in toluene (2.76 ml) and water (0.919 ml). The reaction mixture was heated to 100°C and stirred overnight. The reaction mixture was cooled to room temperature and water was added. The aqueous layer was washed with RINKAN. The combined organic layers were dried over MgSO4 and concentrated. The crude reaction mixture was purified by silica gel chromatography eluting with 0-50% RINKAN / hexane to obtain the title compound (80 mg, 0.343 mmol, 47%).

[0181] Ester hydrolysis was carried out in the same manner as in step d of Example 2. The title compound was isolated by precipitation to obtain the desired product (50 mg, 0.228 mmol, 67%).

[0182] Intermediate 3 [ka] Intermediate 3 step a [ka] A solution of methyl 2-amino-4-methoxybenzo[d]thiazole-6-carboxylate (2 g), CuBr2 (3.7 g, 16.78 mmol), and t-BuNO2 (1.7 g, 16.77 mmol) in CH3CN was stirred at room temperature under an N2 atmosphere for 16 hours. The resulting solution was diluted with water, extracted with SiO2, and the organic layer was dried and concentrated. The resulting solution was purified by silica gel column chromatography (SiO2 in hexane) to obtain the desired product (1.6 g, 63%) as an orange solid. ESI-MS m / z: 301.90 [M+H] + .

[0183] Intermediate 3 Step b [ka] The title compound was synthesized using the compound from step a of intermediate 3 in the same manner as intermediate 1. Ester hydrolysis was carried out in the same manner as in step d of Example 2. The title compound was isolated by precipitation to obtain the desired product (30 mg, 79%).

[0184] Intermediate 4 [ka] The title compound was synthesized using the compound from step a of intermediate 3 in the same manner as intermediate 2. Ester hydrolysis was carried out in the same manner as in step d of Example 2. The title compound was isolated by precipitation to obtain the desired product (57 mg, 75%).

[0185] Intermediate 5 [ka] Intermediate 5 Step a [ka] In a vial, methyl 6-amino-5-bromonicotinate (500 mg, 2.164 mmol) was slurryed in DCM (3.4 ml) and pyridine (2.0 ml). The solution was cooled to 0°C. Cyclopropane carbonyl chloride (590 μl, 6.49 mmol) was added dropwise to homogenize the solution. The reaction mixture was stirred for 2 hours. The reaction stopped upon addition of MeOH, and the solution was then concentrated. MeCN was added, and the reaction mixture was evaporated to remove the pyridine.

[0186] The compound was dissolved in 1:1 THF / MeOH (5 mL) and cooled to 0°C. NaOMe in MeOH (620 μL, 2.7 mmol) was slowly added and the mixture was stirred for 30 minutes. AcOH (250 μL) was added to solidify the reaction. The solid was broken up and water was added. After stirring the solution for a further 10 minutes, the solid was filtered and washed with water to obtain the title compound (546 mg, 1.825 mmol, 84%).

[0187] Intermediate 5 Step b [ka] In a vial, the compound from step a (400 mg, 1.337 mmol) was suspended in THF (5.35 ml), and Lawson's reagent (595 mg, 1.471 mmol) was added. The reaction mixture was heated at 65°C overnight. The reaction mixture was cooled to room temperature, allowed to stand, and then concentrated. The crude reaction mixture was purified by silica gel chromatography eluting with 0-50% HCl / hexane to obtain the title compound (400 mg, 1.27 mmol, 95%).

[0188] Intermediate 5 steps c and d [ka] In a vial, the compound from step b (434 mg, 1.377 mmol) was dissolved in DMSO (3.44 ml). Sodium hydride (60.6 mg, 1.515 mmol) was added, and the reaction mixture was stirred for 5 minutes. The vial was then sealed and heated at 70°C for 5 hours. After cooling, the reaction mixture was diluted with water and extracted with ethyl acetate. The aqueous phase was washed with ethyl acetate. The combined organic phase was washed with brine, dried over MgSO4, filtered, and concentrated. The crude reaction mixture was purified by silica gel chromatography eluting with 0-50% ethyl acetate / hexane to obtain the title compound (130 mg, 0.555 mmol, 40%).

[0189] Ester hydrolysis was carried out in the same manner as in step d of Example 2. The title compound was isolated by precipitation to obtain the desired product (50 mg, 0.232 mmol, 77%).

[0190] Intermediate 6 [ka] Intermediate 6 Step a [ka] To a solution of 5-bromo-1H-indazole (500 mg, 2.54 mmol) in RINKAN (5 mL), 2,2-difluoro-2-(fluorosulfonyl)acetic acid (542 mg, 3.05 mmol) and K2CO3 (701 mg, 5.08 mmol) were added. The resulting mixture was stirred at room temperature for 2 hours and then evaporated. The residue was purified by Combiflash elution with 0-30% RINKAN / hexane to obtain the desired product (380 mg, 60.6%) as a white foam. ESI-MS m / z: 248.9 [M+H] + .

[0191] Intermediate 6 steps b and c [ka] To a solution of DMF / EtOH (6 mL, 2:1), the compound from step a (240 mg, 0.866 mmol), Pd(OAc)2 (23.34 mg, 0.104 mmol), 1,3-bis(diphenylphosphino)propane (86 mg, 0.208 mmol), and triethylamine (362 μl, 2.60 mmol) were added. After degassing, the mixture was subjected to CO balloon filtration. The reaction mixture was heated at 80°C for 12 hours, cooled to room temperature, and diluted with RINKAN (150 mL). The solution was washed with brine, dried, and purified by Combiflash elution with 0-50% RINKAN / hexane to obtain the desired product (130 mg, 55.5%) as a white solid. ESI-MS m / z: 271.2 [M+H] + .

[0192] To a solution of the compound from step b (120 mg, 0.468 mmol) in THF / water (5:1, 6 mL), LiOH (112 mg, 4.68 mmol) was added. The resulting mixture was stirred at 45°C for 20 hours, and then most of the THF was evaporated. The remaining mixture was neutralized to approximately 3 pH by adding 1 M aqueous HCl solution. The precipitated solid was filtered, washed with water, and dried to obtain the desired product (105 mg, 93.0%) as a white solid. ESI-MS m / z: 243.21 [M+H] + .

[0193] Intermediate 7 Steps a and b [ka] To a solution of ethyl 7-chloro-2-(difluoromethyl)-2H-indazole-5-carboxylate (200 mg, 0.728 mmol) in 1,4-dioxane / water (2.1 mL, 20:1), cyclopropylboronic acid (125 mg, 1.456 mmol), XPhos Pd G2 (28.6 mg, 0.036 mmol), and K3PO4 (773 mg, 3.64 mmol) were added. After degassing, the mixture was heated at 80°C for 2 hours to complete the reaction. The reaction mixture was diluted with  (100 mL), washed with brine, dried, and purified by combiflush elution with 0-30%  / hexane to obtain the desired product (93 mg, 45.6%) as a brown solid. ESI-MS m / z: 281.10 [M+H] + .

[0194] To a solution of the compound from step a (90 mg, 0.321 mmol) in THF / water (5:1, 6 mL), LiOH (77 mg, 3.21 mmol) was added. The resulting mixture was stirred at 45°C for 20 hours, and then most of the THF was evaporated. The remaining mixture was neutralized to approximately 3 pH by adding 1 M aqueous HCl solution. The precipitated solid was filtered, washed with water, and dried to obtain the desired product (60 mg, 74.1%) as a brown solid. ESI-MS m / z: 253.20 [M+H] + .

[0195] Intermediate 8 [ka] Intermediate 8 Step a [ka] To a solution of 5-bromo-3-methoxypyridine-2-amine (400 mg, 1.970 mmol) in EtOH (6 mL), 3-bromo-1,1,1-trifluoropropan-2-one (752 mg, 3.94 mmol) was added. The resulting mixture was heated overnight at 80°C. After evaporating the solvent, the residue was purified by Combiflash elution with 0-40% siRNA / hexane to obtain the title compound (300 mg, 51.6%) as a colorless oil. ESI-MS m / z: 296.20 [M+H] + .

[0196] Intermediate 8 steps b and c [ka] The title compound was prepared according to a six-step intermediate procedure. The crude product was purified by Combiflash elution with 0-50% siRNA / hexane to obtain the desired product (200 mg, 68.2%) as a white solid. ESI-MS m / z: 289.17 [M+H] + .

[0197] To a solution of the compound from step b (200 mg, 0.694 mmol) in THF / water (5:1, 6 mL), LiOH (166 mg, 6.94 mmol) was added. The resulting mixture was stirred at room temperature for 4 hours, and then most of the THF was evaporated. The remaining mixture was neutralized to approximately 3 pH by adding 1 M aqueous HCl solution. The precipitated solid was filtered, washed with water, and dried to obtain the desired product (120 mg, 66.5%) as a white solid. ESI-MS m / z: 261.20 [M+H] + .

[0198] Intermediate 9 [ka] Intermediate 9 Step a [ka] To a solution of 5-bromo-3-methoxypyridine-2-amine (2.0 g, 8.75 mmol) in EtOH (30 mL), 2-bromo-1-cyclopropylethane-1-one (1.99 g, 11.82 mmol) was added. The resulting mixture was heated at 80°C for 3 days. After evaporating the solvent, the residue was purified by Combiflash elution with 0-100% siRNA / hexane, followed by 0-5% MeOH / DCM, to obtain the title compound (1.88 g, 85%) as a brown foam. ESI-MS m / z: 268.20 [M+H] + .

[0199] Example 671 Steps b and c [ka] BBr3 (3.74 mL, 3.74 mmol) was slowly added to the solution of step a (500 mg, 1.872 mol) from step a in DCM (10 mL). The resulting mixture was stirred at room temperature for 20 hours, then quenched with water (2 mL). After dilution with DCM (50 mL), the organic layer was separated, dried, and evaporated. The residue was purified by Gilson preparative HPLC (20-90%, MeCN / water, 25 min) to obtain the desired product (40 mg, 8.44%) as a brown foam. ESI-MS m / z: 254.10 [M+H] + .

[0200] To a solution of step b (40 mg, 0.158 mmol) from step b in DMF (2 mL), K2CO3 (43.7 mg, 0.316 mmol) and 2,2-difluoroethyl 4-methylbenzene sulfonate (187 mg, 0.79 mmol) were added. The resulting mixture was heated in a sealed container at 60°C for 20 hours. After evaporating the solvent, the residue was purified by Combiflash elution with 0-30% siRNA / hexane to obtain the desired product (35 mg, 85%) as a pale yellow foam. ESI-MS m / z: 318.99 [M+H] + .

[0201] Intermediate 9 steps d and e [ka] To a solution of DMF / EtOH (6 mL, 2:1), the compound from step c (35 mg, 0.11 mmol), Pd(OAc)2 (2.97 mg, 0.013 mmol), 1,3-bis(diphenylphosphino)propane (10.92 mg, 0.026 mmol), and triethylamine (46 μl, 0.33 mmol) were added. After degassing, the mixture was subjected to CO balloon filtration. The reaction mixture was heated at 80°C for 20 hours, cooled to room temperature, and diluted with HCl (50 mL). The solution was washed with brine, dried, and purified by Combiflash elution with 0-50% HCl / hexane to obtain the desired product (17 mg, 49.6%) as a white solid. ESI-MS m / z: 311.10 [M+H] + .

[0202] To a solution of the compound from step d (17 mg, 0.055 mmol) in THF / water (5:1, 1 mL), 2N NaOH (0.2 mL) was added. The resulting mixture was stirred at room temperature for 40 hours, and then most of the THF was evaporated. The remaining mixture was neutralized to approximately 3 pH by adding 1 M aqueous HCl solution. After extracting the mixture with 10% MeOH / DCM (50 mL x 3), the combined organic layers were combined, dried, and evaporated to obtain the desired product (7 mg, 45.3%) as a white solid. ESI-MS m / z: 283.07 [M+H] + .

[0203] Intermediate 10 [ka] Intermediate 10 Step a [ka] To a solution of 2-amino-5-bromo-3-methoxybenzoic acid (4.00 g, 16.25 mmol) in THF (60 mL), BH3-THF (325 mL, 325.10 mmol) was added dropwise under an ice / water bath, and the reaction mixture was stirred overnight at 50°C. The mixture was cooled to 0°C, quenched with MeOH, and concentrated. The residue was diluted with aqueous Na2CO3 and extracted with EA. The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product (4.4 g) as a yellow oily substance. ESI-MS m / z: 231.95 [M+H] + .

[0204] Intermediate 10 steps b and c [ka] The mixture of the compound from step a (4.40 g, 18.96 mmol) and MnO2 (8.24 g, 94.80 mmol) in DCM (50 mL) was stirred overnight at room temperature. The resulting mixture was filtered, and the filter cake was washed with DCM. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (10:1) to obtain the desired product (2.9 g, 66%) as a red solid. ESI-MS m / z: 229.95 [M+H] + .

[0205] To a solution of the compound from step b (2.90 g, 12.60 mmol) in HCl (6 M) (30 mL) at 0°C, NaNO2 (1.09 g, 15.80 mmol) in water (5 mL) was added. After 30 minutes, the ice bath was removed and the reaction mixture was stirred at room temperature for 30 minutes. The solid was removed by filtration, the mother liquor was cooled to 0°C, and treated with NaN3 (0.82 g, 12.60 mmol) in water (5 mL). The ice bath was removed and stirring was continued for 30 minutes. The resulting solid (3.8 g) was recovered by filtration.

[0206] Intermediate 10 Step d [ka] The compound from step c (3.80 g, 14.84 mmol) and aminocyclopropane (1.27 g, 22.26 mmol) from step c were treated with molecular sieves in toluene (50 mL). The reaction mixture was stirred at room temperature for 2 hours, then heated overnight under reflux. After cooling to room temperature, the mixture was filtered through Celite, and the solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to obtain the desired product (2.4 g, 60%) as a brown oil. ESI-MS m / z: 267.05 [M+H] + .

[0207] Intermediate 10 steps e and f [ka] To a solution of the compound from step d (2.40 g, 8.98 mmol) in EtOH (10 mL), DMF (10 mL), TEA (2.5 mL), Pd(OAc)2 (0.40 g, 1.78 mmol), and dppp (1.48 g, 3.59 mmol) were added in a pressure tank. The mixture was pressurized with carbon monoxide to 15 atm at 100°C overnight. The reaction mixture was cooled to room temperature and concentrated under vacuum. The residue was purified by silica gel column chromatography eluting with PE / EA (2:1) to obtain the desired product (2.2 g, 94%) as a yellow oil. ESI-MS m / z: 261.10 [M+H] + .

[0208] The compound from step e (2.10 g, 8.07 mmol) and MeOH (30 mL) were added to a 100 mL round-bottom flask at room temperature. A solution of LiOH (1.93 g, 80.59 mmol) in H2O (10 mL) was added, and the mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 5 with HCl (2 M aqueous solution). The product was recovered by filtration and washed with water (1.28 g). ESI-MS m / z: 233.05 [M+H] + .

[0209] Intermediate 11 Steps a, b and c [ka] A solution of methyl 1H-indazole-5-carboxylate (3 g, 17.03 mmol) and F-TEDA-BF4 (12 g, 34.06 mmol) in DMF was stirred at 80°C for 16 hours under a nitrogen atmosphere. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product (1.5 g, 45%) as a yellow solid. ESI-MS m / z: 194.95 [M+H] + .

[0210] The solution of the compound from step a (1.5 g, 7.72 mmol), CH3I (2.2 g, 15.45 mmol), and Cs2CO3 (7.6 g, 23.17 mmol) in DMF was stirred at room temperature for 4 hours. The resulting solution was diluted with water, extracted with EA, the organic layer was dried, and concentrated. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product (600 mg, 37%) as a yellow solid. ESI-MS m / z: 209.00 [M+H] + .

[0211] The compound from step b (600 mg, 2.88 mmol) and LiOH (690 mg, 28.82 mmol) were mixed in THF / H2O=10:1 (11 mL) and stirred at room temperature for 16 hours. The resulting mixture was concentrated under vacuum. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product (211.9 mg, 38%) as a pale yellow solid. ESI-MS m / z: 195.05 [M+H] + .

[0212] Intermediate 12 [ka] Intermediate 12 Step a [ka] To a mixture of 4-amino-3-fluoro-5-methylbenzonitrile (1.60 g, 10.65 mmol) in HCl (8 M) (15 mL), NaNO2 (0.77 g, 11.19 mmol) in H2O (2 mL) was added dropwise at -5 °C. The reaction mixture was stirred for 30 minutes, and 2-methyl-2-propantheol (0.96 g, 10.64 mmol) in EtOH (5 mL) was added at 0 °C, and the mixture was stirred for 1 hour. The reaction mixture was quenched with water / ice and extracted with EA. The combined organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (20:1) to obtain the desired product (2.7 g, 99%) as an orange oil. ESI-MS m / z: 252.00 [M+H] + .

[0213] Intermediate 12 Step b [ka] A mixture of the compound from step a (2.70 g, 10.74 mmol) and t-BuOK (9.64 g, 85.91 mmol) in DMSO (25 mL) was stirred at room temperature for 30 minutes. The resulting mixture was diluted with water and extracted with EA. The combined organic layer was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (940 mg, 54%) as a yellow solid.

[0214] Intermediate 12 steps c and d [ka] The mixture of the compound from step b (900 mg, 5.58 mmol), iodoethane (1.05 g, 6.7 mmol), and K2CO3 (1.54 g, 11.17 mmol) in DMF (8 mL) was stirred overnight at room temperature. The resulting mixture was poured into water and extracted with EA. The combined organic layer was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (510 mg, 48%) as a grayish-white solid.

[0215] In a 100 mL round-bottom flask, the compound from step c (510 mg, 2.69 mmol), KOH (2.27 g, 40.46 mmol), EtOH (21 mL), and H2O (7 mL) were added at room temperature. The resulting mixture was stirred overnight at 80 °C. The resulting mixture was concentrated under reduced pressure. The residue was acidified to pH 5 with HCl (2 M aqueous solution). The product was recovered by filtration and washed with water to obtain the title compound (545 mg). ESI-MS m / z: 209.15 [M+H] + .

[0216] Intermediate 13 Steps a and b [ka] A mixture of the compound from intermediate step 12b (900 mg, 5.58 mmol), K2CO3 (1.54 g, 11.14 mmol), and iodoethane (1.05 g, 6.70 mmol) in DMF (8 mL) was stirred overnight at room temperature. The resulting mixture was poured into water and extracted with EA. The combined organic layer was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (280 mg, 26%) as a grayish-white solid.

[0217] In a 100 mL round-bottom flask, the compound from step a (280 mg, 1.48 mmol), KOH (1.25 g, 22.28 mmol), EtOH (21 mL), and H2O (7 mL) were added at room temperature. The resulting mixture was stirred overnight at 80 °C. The resulting mixture was concentrated under reduced pressure. The residue was acidified to pH 5 with HCl (2 M aqueous solution). The product was recovered by filtration and washed with water to obtain the title compound (263 mg). ESI-MS m / z: 209.15 [M+H] + .

[0218] Intermediate 14 [ka] Intermediate 14 Step a [ka] 4-bromo-2-chloro-6-methylpyridine (4.5 g, 22 mmol) and THF (100 mL) were added to a 250 mL three-necked round-bottom flask. The flask was cooled to -60°C, and LDA (4.43 mL, 33 mmol) was added dropwise, and the mixture was stirred for 30 minutes. Then, N-methoxy-N-methylcyclopropanecarboxamide (4.2 g, 33 mmol) was added. The resulting mixture was stirred under a nitrogen atmosphere at -60°C for 1 hour. The reaction was monitored by TLC. The reactants were quenched with saturated NH4Cl (aqueous solution), and the aqueous layer was extracted with ELISA. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 10% to 50% gradient over 25 minutes; detector, UV 254 nm, to obtain the desired compound (2.8 g, 47%) as a grayish-white solid. ESI-MS m / z: 273.95 [M+H] + .

[0219] Intermediate 14 Step b [ka] In a 100 mL round-bottom flask, the compound from step a (2.8 g, 10 mmol), NH2OH HCl (3.5 g, 51 mmol), NaOH (2 g, 51 mmol), and MeOH (30 mL) were added at room temperature. The resulting mixture was stirred overnight at 60°C under a nitrogen atmosphere. The reaction was monitored by LC-MS. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / ethyl acetate (1:1) to obtain the desired compound (2.2 g, 75%) as a grayish-white solid. ESI-MS m / z: 288.95 [M+H] + .

[0220] Intermediate 14 steps c and d [ka] In a 100 mL round-bottom flask, the compound from step b (2.2 g, 8 mmol), TFAA (1.8 g, 8 mmol), TEA (3.8 g, 38 mmol), and DME (15 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. The reaction was monitored by LC-MS. The desired product was detected by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 10% to 50% gradient over 25 minutes; detector, UV 254 nm, to obtain the desired compound (1.8 g, 87%) as a grayish-white solid. ESI-MS m / z: 270.95 [M+H] + .

[0221] In a 40 mL vial, the compound from step c (1.8 g, 7 mmol), ferrous chloride (84 mg, 0.7 mmol), and DME (10 mL) were added at room temperature. The resulting mixture was stirred at 80°C for 2 hours under a nitrogen atmosphere. The reaction was monitored by TLC. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / ethyl acetate (1:1) to obtain the desired compound (1.2 g, 67%) as a grayish-white solid. ESI-MS m / z: 270.95 [M+H] + .

[0222] Intermediate 14 Step e [ka] In a 40 mL vial, the compound from step d (1.2 g, 4.4 mmol), MeONa (716 mg, 13 mmol), and THF (10 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. The reaction was monitored by LC-MS. The desired product was detected by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 10% to 50% gradient over 25 minutes; detector, UV 254 nm, to obtain the desired compound (800 mg, 68%) as a grayish-white solid. ESI-MS m / z: 267.00 [M+H] + .

[0223] Intermediate 14 steps f and g [ka] In a 30 mL pressure tank reactor, the compound from step e (400 mg, 1.5 mmol), Pd(AcO)2 (101 mg, 0.45 mmol), DPPP (371 mg, 0.9 mmol), EtOH (4 mL), Et3N (1 mL), and DMF (4 mL) were added at room temperature. The resulting mixture was stirred overnight at 100 °C under a 10 atm CO atmosphere. The reaction was monitored by TLC. The resulting mixture was filtered, and the filter cake was washed with EtOH. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 10% to 70% gradient over 25 minutes; detector, UV 254 nm, to obtain the desired compound (360 mg, 92%) as a white solid. ESI-MS m / z: 261.10 [M+H] + .

[0224] In a 50 mL round-bottom flask, the compound from step f (360 mg, 1.4 mmol), LiOH (166 mg, 7 mmol), MeOH (5 mL), and H2O (1 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. The reaction was monitored by LC-MS. The mixture was acidified to pH 6 with HCl (aqueous solution). The resulting mixture was concentrated under reduced pressure. The precipitated solid was collected by filtration and washed with water (3 × 3 mL) to obtain the desired compound (280 mg, 87%) as a grayish-white solid. ESI-MS m / z: 233.00 [M+H] + .

[0225] Intermediate 15 [ka] Intermediate 15 Step a [ka] A solution of 5-bromo-3-methoxypyridine-2-amine (5 g, 24.63 mmol) and 2-bromo-1-cyclopropylethanone (8.03 g, 49.26 mmol) in EtOH (20 mL) was stirred overnight at 80°C. The residue was purified by reverse-phase flushing to obtain the desired product (4.4 g, 67%) as a yellow solid. ESI-MS m / z: 267.00 [M+H] + .

[0226] Intermediate 15 steps b and c [ka] A solution of the compound from step a (4.4 g, 16.47 mmol), Pd(OAc)2 (740 mg, 3.29 mmol), and dppp (2.7 g, 6.59 mmol) in DMF (12 mL), EtOH (12 mL), and TEA (3 mL) was stirred overnight at 100°C under a CO atmosphere. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (3 g, 70%) as a yellow solid. ESI-MS m / z: 261.00 [M+H] + .

[0227] The solution of the compound from step b (3 g, 11.53 mmol) and LiOH (2.76 g, 115.25 mmol) in EtOH (10 mL) and H2O (10 mL) was stirred at room temperature for 2 hours. The residue product was purified by reverse-phase flushing to obtain 1.3 g of the desired product as a yellow solid. ESI-MS m / z: 233.00 [M+H] + .

[0228] Intermediate 16 [ka] Intermediate 16 Step a [ka] A solution of 5-bromo-3-methoxypyridine-2-amine (4 g, 19.7 mmol) and methyl 3-bromo-2-oxopropanoate (7 g, 39.4 mmol) in EtOH (10 mL) was stirred at 80°C for 16 hours. The residue product was purified by reverse-phase flushing to obtain the desired product (4.6 g, 82%) as a yellow solid. ESI-MS m / z: 285.00 [M+H] + .

[0229] Intermediate 16 steps b and c [ka] The compound from step a (3 g, 10.5 mmol) and a solution of DIBAL-H (30 mL) in THF (30 mL) were stirred at 0°C for 1 hour. The residue was purified by silica gel column chromatography eluted with DCM / MeOH (10 / 1) to obtain the desired product (1.6 g, 59%) as a yellow solid. ESI-MS m / z: 257.00 [M+H] + .

[0230] The solution of the compound from step b (1 g, 3.9 mmol) and MnO2 (3.4 g, 38.9 mmol) in DCM (20 mL) was stirred overnight at room temperature. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. 600 mg of the crude product was used directly in the next step. ESI-MS m / z: 255.00 [M+H] + .

[0231] Intermediate 16 Step d [ka] The compound (600 mg, 2.4 mmol) from step c and a solution of DAST (1 mL) in DCM (10 mL) were stirred at 0°C for 2 hours. The aqueous layer was extracted with EA. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (400 mg, 61%) as a yellow solid. ESI-MS m / z: 277.00 [M+H] + .

[0232] Intermediate 16 Step e [ka] A solution of the compound from step d (400 mg, 1.4 mmol) and Pd(dppf)Cl2 (211 mg, 0.29 mmol) in DMF (10 mL), H2O (10 mL), and TEA (2 mL) was stirred at 100 °C for 8 hours under CO (15 atm). The residue product was purified by reverse-phase flushing to obtain the desired product (188 mg, 54%) as a yellow solid. ESI-MS m / z: 243.00 [M+H] + .

[0233] Intermediate 17 [ka] Intermediate 17 Step a [ka] To a stirred solution of methyl 4-amino-3-methoxyphenyl carboxylate (500 mg, 2.76 mmol) and 2-bromocyclopenta-1-ene-1-carbaldehyde (1.45 g) in toluene, BINAP (687 mg, 1.10 mmol) and Pd(OAc)2 (124 mg, 0.55 mmol) were added. The reaction mixture was heated at 90°C for 3 hours under a nitrogen atmosphere. The resulting mixture was extracted with siRNA (3 × 0 mL). The combined organic layers were washed with siRNA (3 × 10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / siRNA (1:1) to obtain the desired product (600 mg, 79%) as a yellow oil. ESI-MS m / z: 276.00 [M+H] + .

[0234] Intermediate 17 steps b and c [ka] A solution of the compound from step a (500 mg) and In(TfO)2 (3 g) in xylene (5 mL) was stirred at 130 °C for 12 hours under an N2 atmosphere. The resulting mixture was directly concentrated under reduced pressure for the next step. ESI-MS m / z: 258.00 [M+H] + .

[0235] The solution of the compound from step c (1 g, 3.89 mmol) and LiOH (93 mg, 3.89 mmol) in H2O (10 mL) and MeOH (10 mL) was stirred at room temperature for 12 hours. The crude product was purified by reverse-phase flash to obtain the desired product (184.7 mg) as a white solid. ESI-MS m / z: 244.00 [M+H] + .

[0236] Intermediate 18 [ka] Intermediate 18 Steps a and b [ka] A solution of (E)-2-bromocrotonaldehyde (10 g crude) and methyl 4-amino-3-methoxybenzoate (34 g, 134.24 mmol) in AcOH:HCl (2:3, 50 mL) was stirred at 100°C for 1 hour under a nitrogen atmosphere. The resulting solution was concentrated to obtain the desired product (crude product) as a yellow solid. ESI-MS m / z: 295.90 [M+H] + .

[0237] The compounds from step a, ethyl iodide (21 g, 135.08 mmol) and Cs2CO3 (66 g, 202.62 mmol), were mixed in DMF and stirred overnight at room temperature. The resulting solution was diluted with water, extracted with EA (three times), the organic layer was dried, and the mixture was concentrated. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (1 g) as a yellow solid. ESI-MS m / z: 323.90 [M+H] + .

[0238] Intermediate 18 steps c and d [ka] A solution of the compound from step b (1 g, 3.08 mmol), tricyclohexylphosphine (0.9 g, 3.08 mmol), cyclopropylboronic acid (0.8 g, 9.25 mmol), Pd(OAc)2 (140 mg, 0.62 mmol), and K3PO4 (2 g, 9.25 mmol) in toluene and H2O was stirred overnight at 110°C under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product as a yellow solid. ESI-MS m / z: 286.05 [M+H] + .

[0239] The compound from step c and a solution of LiOH (441 mg, 18.43 mmol) in MeOH (10 mL) and H2O (10 mL) were stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product (428 mg) as a yellow solid. ESI-MS m / z: 258.10 [M+H] + .

[0240] Intermediate 19 Steps a and b [ka] The solution of intermediate 18 (step b) (200 mg, 0.62 mmol), L-proline (142 mg, 1.23 mmol), chlorotrimethylaminyl (117 mg, 1.23 mmol), and Cu2O (176 mg, 1.23 mmol) in EtOH was stirred at 80°C for 16 hours. The resulting solution was diluted with water, extracted with EA (three times), the organic layer was dried, and concentrated. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (70 mg, 41%) as a yellow solid. ESI-MS m / z: 279.95 [M+H] + .

[0241] The compound from step a (70 mg, 0.25 mmol) and LiOH (60 mg, 2.50 mmol) were mixed in THF (10 mL) and H2O (1 mL) and stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The resulting solution was purified by reverse-phase C18 column chromatography (MeCN / H2O) to obtain the desired product (50 mg, 79%) as a yellow solid. ESI-MS m / z: 251.95 [M+H] + .

[0242] Intermediate 20 Steps a and b [ka] The solution of intermediate 18 step b (200 mg, 0.62 mmol) in NMP, methyl 2,2-difluoro-2-sulfoacetate (237 mg, 1.23 mmol), KF (72 mg, 1.23 mmol), and CuI (235 mg, 1.23 mmol) was stirred at 120°C for 4 hours. The resulting solution was diluted with water, extracted with EA (3 times), the organic layer was dried, and concentrated. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (140 mg, 72%) as a yellow solid. ESI-MS m / z: 313.95 [M+H] + .

[0243] The compound from step a (140 mg, 0.45 mmol) and LiOH (107 mg, 4.47 mmol) were mixed in MeOH (10 mL) and H2O (1 mL) and stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The resulting solution was purified by reverse-phase C18 column chromatography (CH3CN / H2O) to obtain the desired product (70 mg, 55%) as a white solid. ESI-MS m / z: 285.90 [M+H] + .

[0244] Intermediate 21 [ka] Intermediate 21 Step a [ka] To a stirred solution of methyl 4-amino-3-iodobenzoate (2.7 g, 10 mmol) in HCl (6 mL), NaNO2 (0.7 g in 5 mL of water) was added dropwise at 5°C for 1 hour. Piperidine (1 mL) was added dropwise to the mixture at 5°C. The resulting mixture was stirred at room temperature for a further 1 hour. The resulting mixture was extracted with EA, the combined organic layers were washed with water, and dried over anhydrous Na2SO4. The residue was purified by silica gel column chromatography (hexane containing SiO2) to obtain the desired product (2.7 g) as a yellow solid. ESI-MS m / z: 374.00 [M+H] + .

[0245] Intermediate 21 Step b [ka] Bromo(propa-1-in-1-yl)magnesium (4.3 g, 29.94 mmol) was added to a dry N2 flush 50 mL Schlenk tube equipped with a magnetic stirrer and septum. The solution was cooled to -30°C, and ZnBr2 (5.08 g, 22.56 mmol) was added dropwise to the reaction mixture. The reaction mixture was heated to room temperature for 30 minutes. The compound from step a (2 g, 5.36 mmol) was added, followed by (PPh3)4 (309 mg, 0.27 mmol). The reaction mixture was stirred at room temperature for 2 hours and quenched with saturated NH4Cl aqueous solution. The aqueous layer was extracted with ethyl acetate, dried, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane containing ethyl acetate) to obtain the desired product (2 g, 97%) as a yellow solid. ESI-MS m / z: 286.00 [M+H] + .

[0246] Intermediate 21 Step c [ka] The compound from step b (1.5 g, 5.26 mmol) and an aqueous solution of HBr (850 mg, 10.51 mmol) in acetone (10 mL) were stirred at room temperature for 2 hours. The resulting mixture was extracted with ethyl acetate, the combined organic layer was washed with water, and dried over anhydrous sodium 2SO4. The residue was purified by silica gel column chromatography (hexane containing ethyl acetate) to obtain the desired product (900 mg, 61%) as a yellow solid. ESI-MS m / z: 281.00 [M+H] + .

[0247] Intermediate 21 Step d [ka] The solution of the compound from step c (900 mg, 3.2 mmol) and Pd / C (681 mg, 6.40 mmol) in MeOH (20 mL) was stirred at room temperature under an H2 atmosphere for 2 hours. The resulting mixture was filtered, and the solution was concentrated and used directly in the next step. ESI-MS m / z: 205.00 [M+H] + .

[0248] Intermediate 21 Steps e and f [ka] The compound from step d and a solution of MnO2 (1.5 g, 17.67 mmol) in THF (20 mL) were stirred at room temperature for 2 hours. The crude product was purified by reverse-phase flushing to obtain the desired product (253 mg, 51%) as a yellow solid. ESI-MS m / z: 203.00 [M+H] + .

[0249] In a vial, the compound from step e (100 mg, 0.495 mmol) and lithium hydroxide (118 mg, 4.95 mmol) were dissolved in THF (2.2 ml), MeOH (2.2 ml), and water (0.55 ml). The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water, and the pH was adjusted to 3-4 with 1 M HCl aqueous solution. The aqueous layer was washed with DCM and 9:1 DCM / MeOH. The combined organic layers were dried over MgSO4 and concentrated under reduced pressure to obtain the title compound (45 mg, 48%). ESI-MS m / z: 188.68 [M+H] + .

[0250] Intermediate 21a [ka] Intermediate 21a Step a [ka] The following example was prepared using methyl 4-amino-3-iodo-5-methoxybenzoate (4 g, 13.03) in a procedure similar to step a of intermediate 21 to obtain the title compound. (3.5 g) ESI-MS m / z: 404.00 [M+H] + .

[0251] Intermediate 21a Step b [ka] The following example was prepared using cyclopropylacetylene in a procedure similar to step b of intermediate 21. The title compound was purified by silica gel chromatography to obtain the title compound as a brown oil (3.6 g). ESI-MS m / z: 342.00 [M+H] + .

[0252] Intermediate 21a Step c [ka] The following example was prepared using a procedure similar to step c of intermediate 21 to obtain the title compound (600 mg). ESI-MS m / z: 336.00 [M+H] + .

[0253] Intermediate 21a Step d [ka] The following example was prepared using a procedure similar to step d of intermediate 21 to obtain the title compound (500 mg). ESI-MS m / z: 261.00 [M+H] + .

[0254] Intermediate 21a Steps e and f [ka] The following example was prepared using a procedure similar to steps e and f of intermediate 21 to obtain the title compound (200 mg, 60%). ESI-MS m / z: 245.00 [M+H] + .

[0255] Intermediate 22 [ka] Intermediate 22 Step a [ka] A stirring solution of 2-amino-5-bromo-3-methoxybenzaldehyde (2.2 g, 9.56 mmol), piperazine (2.5 g, 28.69 mmol), and methyl acetacetate (2.2 g, 19.13 mmol) in MeOH was stirred at room temperature for 1 hour. The resulting solution was diluted with water, extracted with EA (three times), and the organic layer was dried and concentrated. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (2.5 g, 84%) as a yellow solid. ESI-MS m / z: 309.85 [M+H] + .

[0256] Intermediate 22 steps b, c and d [ka] A solution of the compound (1 g, 28.69 mmol) from step a in THF was treated with DIBAL-H (20 mL) under a nitrogen atmosphere at room temperature for 1 hour. The resulting solution was diluted with water, extracted with EA (3 times), the organic layer was dried, and concentrated. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (crude product 1 g) as a yellow solid. ESI-MS m / z: 281.90 [M+H] + .

[0257] A solution of the compound from step b (1 g crude) in EtOH and DMF, Pd(OAc)2 (159 mg, 0.71 mmol), DPPP (585 mg, 1.42 mmol), and TEA (2.00 mL) was stirred at 100°C for 16 hours under a CO atmosphere (15 atm). The resulting mixture was concentrated under vacuum. The resulting solution was purified by silica gel column chromatography (PE:EA) to obtain the desired product (600 mg crude product) as a yellow solid. ESI-MS m / z: 276.10 [M+H] + .

[0258] The compound from step c (600 mg crude) and LiOH (522 mg, 21.79 mmol) were mixed in MeOH / H2O=10:1 (11 mL) and stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The resulting solution was purified by reverse-phase C18 column chromatography (MeCN / H2O) to obtain the desired product (504.7 mg) as a pale yellow solid. ESI-MS m / z: 247.95 [M+H] + .

[0259] Intermediate 23 [ka] Intermediate 23 Step a [ka] A solution of methyl 4-amino-3-hydroxybenzoate (5 g, 29.91 mmol) and NCS (4.8 g, 35.89 mmol) in CAN (20 mL) was stirred at 80°C for 5 hours. The residue was purified by silica gel column chromatography eluted with PE / EA (10%) to obtain the desired product (1.2 g) as a yellow solid. ESI-MS m / z: 202.00 [M+H] + .

[0260] Intermediate 23 Steps b and c [ka] The compound from step a (1.2 g, 5.95 mmol) and BrCN (3.2 g, 29.74 mmol) were mixed in MeOH (42 mL) and H2O (18 mL) and stirred at 50°C for 48 hours. The resulting mixture was filtered, and the filter cake was washed with EA and water to obtain the desired product (700 mg, 52%). ESI-MS m / z: 227.00 [M+H] + .

[0261] The compound from step b (500 mg, 2.21 mmol) and LiOH (264 mg, 11.03 mmol) in THF (10 mL) and H2O (10 mL) were stirred at room temperature for 1 hour. The mixture was acidified to pH 7 with HCl (1 M). The crude product was recrystallized from the solution to obtain the desired product (231 mg, 49%) as a brown solid. ESI-MS m / z: 213.00 [M+H] + .

[0262] Intermediate 24 [ka] Intermediate 24 Step a [ka] A mixture of 5-bromo-3-chlorobenzene-1,2-diamine (2.5 g, 11.29 mmol), cyclopropanecarboxylic acid (1.26 g, 14.64 mmol), HATU (4.29 g, 11.29 mmol), DIEA (2.9 g, 22.58 mmol), and DMF (50 mL) was stirred overnight at room temperature. The reaction was monitored by LC-MS. The resulting mixture was poured into water and extracted with SiO2. The combined organic layer was washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to obtain the desired product (6.7 g, crude) as a brown oil. ESI-MS m / z: 289.00 [M+H] + .

[0263] Intermediate 24 steps b and c [ka] A solution of the compound from step a (6.7 g, 23.14 mmol) and acetic acid (60 mL) was stirred overnight at 100°C. The resulting mixture was concentrated under vacuum. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, gradient from 0% to 100% over 30 minutes; detector, UV 254 nm, to obtain the desired product (2 g, 32%) as a yellow solid. ESI-MS m / z: 271.00 [M+H] + .

[0264] To a stirred solution of the compound from step b (2 g, 7.37 mmol) in DMF (20 mL), NaH (0.35 g, 14.73 mmol) was added in fractions at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at the same temperature under a nitrogen atmosphere for 30 minutes. SEM-Cl (2.46 g, 14.76 mmol) was added dropwise, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with water / ice at room temperature. The resulting mixture was extracted with siRNA. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to obtain the desired product (2.6 g, 88%) as a yellow oil. ESI-MS m / z: 401.00 [M+H] + .

[0265] Intermediate 24 steps d and e [ka] To a solution of the compound from step c (2.6 g, 6.47 mmol) in DMF (10 mL), H2O (10 mL), and TEA (2.5 mL), DPPP (1 g, 2.59 mmol) and Pd(OAc)2 (0.29 g, 1.29 mmol) were added in a pressure tank. The mixture was pressurized to 15 atm with carbon monoxide and stirred at 100°C for 2 days. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions to obtain the desired crude product (1.6 g, 67%) as a yellow solid: column, C18 silica gel; mobile phase, MeCN in water (0.05% FA), gradient from 10% to 80% over 25 minutes; detector, UV 254 nm. ESI-MS m / z: 367.10 [M+H] + .

[0266] The compound from step d (1.6 g, 4.4 mmol) and a solution of HCl (6 M, 5 mL) in EtOH (6 mL) and H2O (6 mL) were stirred at 80°C for 16 hours. The crude product was purified by reverse-phase flash to obtain the desired product (300 mg, 29%) as a white solid. ESI-MS m / z: 237.00 [M+H] + .

[0267] Intermediate 25 Steps a and b [ka] A solution of lithium chloride (0.5 M in THF) (10.00 ml, 5.00 mmol) and methyl-d3-magnesium iodide (1.0 M in diethyl ether) (5.00 ml, 5.00 mmol) was treated with zinc chloride (1.9 M in 2-MeTHF) (1.316 ml, 2.500 mmol) at room temperature. A solution of IPr PEPPSI (0.017 g, 0.025 mmol) and methyl 7-bromo-1H-indazole-5-carboxylate (0.128 g, 0.500 mmol) dissolved in 1.5 mL of NMP was added dropwise, and the resulting reaction mixture was stirred overnight at room temperature. Once complete, the reaction mixture was poured into a 5% aqueous citric acid solution and extracted with siRNA. The organic extract was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The title compound (84 mg, 0.435 mmol, 87% yield) was obtained as a grayish-white solid by purification using silica gel flash column chromatography. ESI-MS m / z: 193.7 [M+H] + .

[0268] The solution of step a (0.083 g, 0.430 mmol) in THF (5 ml) was treated with potassium trimethylsilanolate (0.276 g, 2.148 mmol) and stirred at room temperature. Upon completion, the reaction mixture was quenched with MeOH and silica gel, concentrated, and the resulting free-flowing mixture was directly purified by flash column chromatography on silica gel to obtain the title compound as a white solid (0.075 g, 97% yield). ESI-MS m / z: 179.6 [M+H] + .

[0269] Intermediate 26 [ka] Intermediate 26 Step a [ka] Selenium dioxide (183 mg, 1.654 mmol) and 2-methylbenzo[d]thiazole-6-carboxylic acid (213 mg, 1.102 mmol) were dissolved in 1,4-dioxane (2.205 ml) at room temperature. The reaction mixture was heated at 90°C for 16 hours. The mixture was diluted with ethyl acetate and passed through a short silica gel plug. The plug was washed with ethyl acetate, and the resulting solution was concentrated and used as the crude compound. ESI-MS m / z: 207.61 [M+H] + .

[0270] Intermediate 26 Step b [ka] Step a (37 mg, 0.179 mmol) was dissolved in DCM (1 ml), and DAST (83 μl, 0.625 mmol) was added at 0°C. The reaction mixture was stirred at 0°C for 2 hours. The reaction mixture was diluted with ethyl acetate and poured into a 1N HCl solution. The aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried and purified by flash chromatography to obtain the title compound. (15 mg, yield 36.6%) ESI-MS m / z: 230.00 [M+H] + .

[0271] Intermediate 27 Steps a and b [ka] A solution of lithium chloride (0.5 M in THF) (6.17 ml, 3.08 mmol) and methyl-d3-magnesium iodide (1.0 M in diethyl ether) (3.08 ml, 3.08 mmol) was treated with zinc chloride (1.9 M in 2-MeTHF) (0.812 ml, 1.542 mmol) at room temperature. A solution of IPr PEPPSI (10.51 mg, 0.015 mmol) dissolved in 1.5 mL of NMP and a solution of ethyl 3-bromo-8-methoxy-2-methylquinoline-6-carboxylate (0.1 g, 0.308 mmol) was added dropwise, and the resulting reaction mixture was stirred overnight at room temperature. Once complete, the reaction mixture was poured into a 5% aqueous citrate solution and extracted with RINKAN. The organic extract was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by flash chromatography yielded the desired product (50 mg, yield 61.8%) as a yellow solid. ESI-MS m / z:263.02 [M+H] + .

[0272] Ester hydrolysis was carried out according to step d of Example 2 to obtain the desired compound.

[0273] Example 3 [ka] Example 3 Process a [ka] To a solution of (R)-3-cyclopropoxy-4-(2-hydroxypropoxy)benzoic acid (100 mg, 0.396 mmol) in THF (1.982 ml), 4-methylmorpholine (47.9 μl, 0.436 mmol) and tert-butylchlorodimethylsilane (65.7 mg, 0.436 mmol) were added. After stirring the reaction mixture for 30 minutes, 1H-tetrazole (4405 μl, 1.982 mmol) and dibenzyldiisopropylphosphorumidite (400 μl, 1.189 mmol) were added. The reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was cooled to 0°C, and then hydrogen peroxide (121 μl, 1.189 mmol) was added. The reaction mixture was slowly warmed to room temperature over 30 minutes. Then saturated sodium sulfite was added. After vigorously stirring for 30 minutes, the mixture was partitioned between ethyl acetate and saturated sodium sulfite. The pH of the combined aqueous layer was adjusted to 3 with 1N HCl, and the resulting suspension was extracted with ethyl acetate. The combined organic layer was dried over MgSO4. After removing the solvent, the residue was purified by flash chromatography to obtain the title compound (183 mg, 90% yield).

[0274] Example 3: Steps b and c [ka] The amide coupling was carried out according to step c of Example 1 to obtain the desired compound.

[0275] Step b (193 mg, 0.216 mmol) was dissolved in ethyl acetate (2.159 ml) in a 40 mL vial. The vial was flushed with nitrogen gas three times. Palladium carbon (22.98 mg, 0.022 mmol) was added all at once. The vial was flushed with a hydrogen balloon, and the reaction mixture was stirred at 25°C for 2 hours. The hydrogen gas was removed, and nitrogen was flushed. The solution was filtered through celite, the residue was concentrated, and purified by HPLC. ESI-MS m / z: 714.13 [M+H] + .

[0276] Example 4 [ka] Example 4 Process a [ka] The following examples were prepared according to step c of Example 1, using the acid coupling partner 2-formyl-8-methoxy-3-methylquinoline-6-carboxylic acid (55 mg).

[0277] Example 4 Step b [ka] The product from step a (36 mg, 0.057 mmol) was dissolved in EtOH, and sodium borohydride (4.35 mg, 0.115 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was quenched by carefully adding saturated NH4Cl. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were dried and purified by HPLC to obtain the title compound (14.5 mg, 40.1% yield). ESI-MS m / z: 629.27 [M+H] + .

[0278] Example 5 [ka] Next, step a of Example 4 (38 mg, 0.061 mmol) was dissolved in tBuOH and water. 2-Methyl-2-butene (129 μl, 1.213 mmol) and sodium dihydrogen phosphate (72.8 mg, 0.606 mmol) were added at room temperature. Then, sodium chlorite (68.6 mg, 0.606 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. The reaction was quenched by adding a 15% Na2S2O3 solution, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were dried and purified by HPLC to obtain the title compound (14 mg, 0.022 mmol, yield 35.9%). ESI-MS m / z: 643.15 [M+H] + .

[0279] Intermediate 28 Steps a and b [ka] Methyl 4-amino-3-methoxybenzoate (2.86 g, 15.79 mmol) and (E)-2-ethylbuta-2-enal (5.468 g, 18.94 mmol) were added to a 100 mL rbf container equipped with a stirring bar, followed by AcOH (23.68 ml) and HCl (15.79 ml). The mixture was then heated at 100 °C for 2 hours. The mixture was then cooled to room temperature and partially concentrated under vacuum at 60 °C to obtain a concentrated black oil. This oil was treated with 40 mL of EtOH and 250 μL of sulfuric acid, and then heated at 80 °C for 2 hours. The mixture was cooled to room temperature and then neutralized and concentrated by the addition of 2 mL of NEt3. The resulting residue was purified by automated silica gel column chromatography (0-45% ethyl acetate in hexane) and dried under high vacuum to obtain the title compound as a colorless oil (49.8 mg, 1%). ESI-MS m / z:274.2 [M+H] + .

[0280] Ester hydrolysis was carried out in the same manner as in step d of Example 2. The crude product was allowed to proceed as a white solid without further purification. ESI-MS m / z: 246.0 [M+H] + .

[0281] Intermediate 29 [ka] In a 4 mL vial, 125 mg (0.714 mmol) of 2-methyl-1H-indole-5-carboxylic acid was added, followed by 1.427 mL of DMF. The mixture was cooled to 0°C, and then NaH (86 mg, 2.141 mmol) was slowly added. The mixture was stirred for 15 minutes, and then methyl iodide (134 μL, 2.141 mmol) was added. After 2 hours, the mixture was neutralized by adding 1 M HCl (aqueous solution) and then diluted with 2 mL of dichloromethane. The layers were separated, and the aqueous layer was washed with 5 × 1 mL of dichloromethane. The combined organic layers were washed with water, dried over Na₂SO₄, filtered, concentrated, and then purified by automated silica gel column chromatography. The mixture was dried under high vacuum to obtain the title compound as a white solid (65 mg, 38%). ESI-MS m / z: 189.7 [M+H] + .

[0282] Intermediate 30 [ka] Intermediate 30 Step a [ka] 3-methyl-1H-pyrazole-5-amine (1.00 g, 10.3 mmol), AcOH (20.6 ml), and diethylbuta-2-indioate (1.65 ml, 10.3 mmol) were added to a 20 mL vial. The mixture was stirred at 23 °C for 48 hours, then poured into hexane:siRNA (1:1, 100 mL) and stirred for 1 hour. The solid was recovered by filtration to obtain the title compound (1.78 g, 78%) as a yellow solid. ESI-MS m / z: 221.9 [M+H] + .

[0283] Intermediate 30 steps b and c [ka] In a 20 mL vial, step a (250 mg, 1.13 mmol), cesium carbonate (736 mg, 2.26 mmol), and DMF (2.3 mL) were added, followed by the addition of MeI (212 μl, 3.39 mmol). The mixture was stirred for 16 hours. The mixture was then diluted with 5 mL of water and 5 mL of dichloromethane. The layers were separated, and the aqueous layer was washed with 5 × 5 mL of dichloromethane. The combined organic layers were washed with 2 × 1 mL of water, then with 1 mL of brine, dried over Na₂SO₄, concentrated, and purified by automated silica gel chromatography to obtain the title compound as a yellow oil (205 mg, 77%). ESI-MS m / z: 235.9 [M+H] + .

[0284] Ester hydrolysis was carried out in the same manner as in step d of Example 2. After acidification, the product was recovered by filtration and proceeded as a yellow solid without further purification. ESI-MS m / z: 207.7 [M+H] + .

[0285] Intermediate 31 [ka] In a 20 mL vial, 6-methylnicotinic acid (333 mg, 2.43 mmol) and 2-bromo-1-cyclopropylethane-1-one (396 mg, 2.43 mmol), followed by acetonitrile (12.0 mL), were added and heated at 70°C for 16 hours. Then, triethylamine (1.35 mL, 9.71 mmol) was added, and the mixture was stirred for a further 3 hours. The mixture was then diluted with 5 mL of water and 5 mL of toluene. The layers were separated, and the aqueous layer was washed with 4 × 5 mL of toluene. The combined organic layers were washed with 2 mL of brine, dried over Na₂SO₄, filtered, concentrated, and purified by automated silica gel chromatography to obtain the title compound (78 mg, 16%). ESI-MS m / z: 201.8 [M+H] + .

[0286] Intermediate 32 Steps a and b [ka] 6-bromo-2-cyclopropylimidazo[1,2-b]pyridazine (128 mg, 0.538 mmol), palladium(II) acetate (12 mg, 0.054 mmol), and 1,3-bis(diphenylphosphino)propane (44.3 mg, 0.108 mmol) were placed in an 8 mL vial, and the headspace of the vial was purged with N2 for 30 minutes. A mixture of DMF (1.434 ml), ethanol (0.717 ml), and triethylamine (225 μl, 1.613 mmol) was degassed by spraying with N2 for 30 minutes, and then added to the solid mixture. The vial was placed under a CO balloon, and the mixture was heated at 80°C for 16 hours. Additional palladium(II) acetate (12 mg, 0.054 mmol) was added, and the mixture was heated at 100°C for 4 hours. The reaction vessel was degassed by sparging with N2, and the mixture was then concentrated and purified by automated silica gel chromatography to obtain the title compound as a yellow oil (35 mg, 28%). ESI-MS m / z: 232.0 [M+H] + .

[0287] Ester hydrolysis was carried out in the same manner as in step d of Example 2. The product could not be extracted from the aqueous layer. The aqueous layer was concentrated to obtain the title compound as a mixture with lithium chloride. ESI-MS m / z: 204.1 [M+H] + .

[0288] Intermediate 33 [ka] Intermediate 33 Step a [ka] 1H-pyrrole-1-amine (1 ml, 13.15 mmol) and its diethyl 2-(ethoxymethylene)malonate (3.11 ml, 15.39 mmol) were added to a 20 mL vial and heated at 125°C for 1 hour. The mixture was diluted with a mixture of biphenyl (1 g) and diphenyl ether (3 ml) and then heated at 200°C for 2 hours. The mixture was cooled to room temperature and then purified by automated silica gel chromatography (0-10% ethyl acetate in cyclohexane) to obtain the title compound as a yellow solid (1.43 g, 53%). ESI-MS m / z: 207.1 [M+H] + .

[0289] Intermediate 33 Step b [ka] In an 8 mL vial, the compound from step a (300 mg, 1.46 mmol), CCl4 (1.5 mL), celite (150 mg), and triphenylphosphine (1.13 g, 4.36 mmol) were added. The mixture was heated at 75 °C for 16 hours. The reaction product was diluted with SiO2 and filtered through celite. The filtrate was concentrated and purified by automated silica gel chromatography to obtain the expected ester (243 mg, 74%) and hydrolysis product as a fluorescent yellow solid (19 mg, 7%), which was then directly incubated. ESI-MS m / z: 197.0 / 199.0 [M+H] + .

[0290] Intermediate 34 [ka] In an 8 mL vial, the ester (131 mg, 0.583 mmol) from intermediate step 33b and ammonium formate (184 mg, 2.92 mmol) were added. A mixture of dioxane (2.92 ml) and ethanol (2.92 ml) was degassed by sparging with N2 for 30 minutes and then added to the mixture. Palladium carbon (62.1 mg, 0.583 mmol, 10 wt%) was added, the mixture was stirred for 1 hour, and then filtered through celite. The filtrate was concentrated and purified by automated silica gel chromatography to obtain the product as a fluorescent yellow solid (77 mg, 70%). ESI-MS m / z: 191.1 [M+H] + .

[0291] Ester hydrolysis was carried out in the same manner as in step d of Example 2. After acidification, the title compound was recovered as a fluorescent yellow solid by filtration (19 mg, 32%). ESI-MS m / z: 163.0 [M+H] + .

[0292] Intermediate 35 Steps a and b [ka] Methyl 4-amino-3-bromobenzoate (345 mg, 1,500 mmol), Pd(PPh3)2Cl2 (211 mg, 0,300 mmol), and copper(I) iodide (28.6 mg, 0.150 mmol) were added to an 8 mL vial. The reaction vessel was evacuated and refilled with N2 three times. Toluene (2 mL) and water (1 mL) were degassed by sparging with N2 for 15 minutes, and then added to the vial together with the solid. Ethynylcyclopropane (508 μL, 6.00 mmol) and triethylamine (627 μL, 4.50 mmol) were added, and the mixture was stirred at 70°C for 16 hours. The mixture was then diluted with water and ethyl acetate, the phases were separated, the organic layer was concentrated, and purified by automated silica gel chromatography (0-10% ethyl acetate in cyclohexane). The title compound was obtained as a colorless oil (298 mg, 92%). ESI-MS m / z:216.1 [M+H] + .

[0293] Dimethyl sulfoxide (1 ml) and KOtBu (155 mg, 1.38 mmol) were added to an 8 mL vial containing the compound from step a (99 mg, 0.46 mmol), and the mixture was then heated to 100 °C for 30 minutes. The mixture was cooled to room temperature, diluted with water and HCl, and then acidified to pH 3 with 1 M (aqueous solution) HCl. The layers were separated, and the aqueous layer was washed with 4 × 3 mL of HCl. The combined organic layers were then washed twice with water and then brine, dried over Na₂SO₄, and purified by automated silica gel chromatography (0-50% in cyclohexane (1% AcOH in HCl)) to obtain the title compound as a white solid (45 mg, 49%). ESI-MS m / z: 202.1 [M+H] + .

[0294] Intermediate 36 [ka] Intermediate 36 Step a [ka] In a 100 mL round-bottom flask, Example 1, step b (3.80 g, 10.27 mmol), and acetone (100 mL) were added, the solution was cooled to 0°C, and then Jones' reagent (1.9-2.2 M, 10 mL) was added dropwise (with internal temperature monitoring). The reaction mixture was warmed to room temperature and monitored by LC-MS (3 hours). The reaction mixture was cooled to 0°C, i The reaction was quenched with PrOH and stirred for 15 minutes. The reaction mixture was diluted with RINKAN and water. The aqueous layer was extracted, and the combined organic layers were dried and concentrated under reduced pressure to obtain the crude product as a yellow solid (3.95 g, 99%). ESI-MS m / z: 383.80 [M+H] + .

[0295] Intermediate 36 Step b [ka] In a 100 mL round-bottom flask, the compound from step a (3.95 g, 10.28 mmol) and NH4Cl (1.10 g, 20.57 mmol) were added, and the solid was dissolved in DMF (20 mL). Hünig base (5.27 mL, 30.84 mmol) was added, the reaction mixture was cooled to 0°C, and HATU (7.82 g, 20.56 mmol) was added. The reaction mixture was warmed to room temperature and monitored by LC-MS (1 hour). The reaction mixture was diluted with HCl and water. The aqueous layer was extracted, the combined organic layers were dried, and concentrated. The substance was purified by automated column chromatography (silica gel, hexane, 0-100% HCl) to obtain the title compound (3 g, 76%). ESI-MS m / z: 382.95 [M+H] + .

[0296] Intermediate 36 Step c [ka] In a 100 mL round-bottom flask, the compound from step b (3.00 g, 7.83 mmol), 3,3,3-trifluoropropane-1-en-2-ylboronic acid (2.19 g, 15.65 mmol), and Pd(dppf)Cl2 (1.15 g, 1.56 mmol) were added, and the material was dissolved in dioxane (40 mL) and H2O (5 mL). Then, K2CO3 (3.25 g, 23.50 mmol) was added, and the resulting mixture was stirred at 90°C for 1 hour under a nitrogen atmosphere. The mixture was cooled to room temperature, poured into water, extracted with ethyl acetate, and the combined organic layers were concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, hexane, 0-100% ethyl acetate) to obtain the desired product as a brown oil (2.3 g, 83%). ESI-MS m / z: 350.90 [M+H] + .

[0297] Intermediate 36 Step d [ka] To a stirred solution of step c (5.00 g, 14.24 mmol) and 3-chloro-4-fluorophenylboronic acid (3.72 g, 21.33 mmol) in THF (80 mL), Na2CO3 (3.32 g, 31.33 mmol), H2O (20 mL), and Pd(PPh3)2Cl2 (1.00 g, 1.42 mmol) were added. The resulting mixture was stirred at 70°C for 1 hour under a nitrogen atmosphere. The reaction was monitored by TLC and LCMS. The resulting mixture was extracted with HCl, the combined organic layers were washed with brine, dried, and concentrated under reduced pressure. The residue was purified by automated column chromatography (silica gel, hexane with 0-75% HCl) to obtain the title compound as a yellow solid (5.8 g, 99%). ESI-MS m / z: 401.05 [M+H] + .

[0298] Intermediate 36 Step e [ka] AD-Mix-β (33.82 g, 43.41 mmol) and methanesulfonamide (1.38 g, 14.47 mmol) were added to a 500 mL round-bottom flask equipped with a stirring bar. The solid was dissolved in tBuOH (60 mL) and H2O (100 mL), the flask was cooled to 0°C, and the compound from step d (5.80 g, 14.47 mmol) was slowly added as a solution in tBuOH (40 mL). The reaction mixture was allowed to warm naturally to room temperature and stirred for 16 hours. The reaction was quenched by adding sodium sulfite (0.25 g per 1 g of AD-Mix), and the mixture was diluted with water and SiO2. The layers were separated, and the aqueous layer was extracted with SiO2. The combined organic layers were washed with brine, dried over Na2SO4, filtered, concentrated, and purified by automated column chromatography (silica gel, hexane, 0-100% SiO2) to obtain the title compound as a white solid (5.48 g, 87%). ESI-MS m / z:435.[M+H] + .

[0299] Intermediate 36 Step f [ka] In a 250 mL round-bottom flask, the compound from step e (4.70 g, 10.81 mmol) and DCM (80 mL) were added at room temperature. The solution was cooled to 0°C, and then DMAP (264 mg, 2.16 mmol), TEA (3.28 g, 32.43 mmol), and TsCl (2.47 g, 12.97 mmol) were added sequentially. The resulting mixture was stirred at 0°C for 1 hour. The mixture was acidified to pH 4 with 2 M HCl, and the aqueous phase was extracted with DCM. The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain the crude product as a pale yellow solid (6.2 g, 97%). ESI-MS m / z: 589.15 [M+H] + .

[0300] Intermediate 36 Step g [ka] In a 100 mL round-bottom flask, NH3 in MeOH (35 mL) was added at room temperature, and the compound from step g (6.20 g, 10.52 mmol) was slowly added. The resulting mixture was stirred at room temperature and monitored by LC-MS (5 hours). The mixture was dissolved in ethyl acetate, washed three times with saturated sodium bicarbonate, washed with brine, dried, and concentrated to obtain the title compound (2.93 g, 64%). ESI-MS m / z: 434.05 [M+H] + .

[0301] Intermediate 37 [ka] Intermediate 37 Step a [ka] N-fluoro-N-(phenylsulfonyl)benzenesulfonamide (963 mg, 3.05 mmol) and methyl 7-bromo-5-iodo-2,3-dihydrofluoro[2,3-c]pyridine-3-carboxylate (586 mg, 1.526 mmol) were added to a 20 mL vial equipped with a stirring bar, and the solid was dissolved in THF (0.25 M). The vial was cooled to -78°C, and LDA (916 μl, 1.831 mmol) was slowly added. The reaction mixture was stirred at -78°C for 1 hour. After 1 hour at -78°C, the reaction mixture gelled. The reaction mixture was warmed to room temperature, stirred, and monitored by LC-MS. The reaction mixture was diluted with  and quenched with water and saturated ammonium chloride.  and DCM / MeOH were extracted once each using a phase separator cartridge. Concentration. The material was purified by automated column chromatography (0-100% Âx / cHex) to obtain the title compound (483 mg, 71%). ESI-MS m / z: 401.86 / 403.86 [M+H] + .

[0302] Intermediate 37 Step b [ka] The material was added to a 20 mL vial containing step a (483 mg, 1.081 mmol) and stirred with a stirring bar to dissolve the material in THF / MeOH and water (1:1:1, 0.25 M). Lithium hydroxide hydrate (113 mg, 2.70 mmol) was added, and the reaction was stirred at room temperature and monitored by LC-MS.

[0303] The reaction mixture was diluted with RINKAN and acidified to pH=2 with 2M HCl. Extraction with RINKAN and DCM / MeOH using a phase separator cartridge was performed. This substance was triturated with DCM / hexane to obtain an orange solid. (455 mg, 92%) ESI-MS m / z: 387.82 / 387.82 [M+H] + .

[0304] Intermediate 37 Step c [ka] The above compound was prepared in a manner similar to step b of intermediate 36. The crude substance was purified by automated column chromatography (silica gel, 0-100% HCl / cHex) to obtain the title compound as an orange oily substance (243 mg, 43%). ESI-MS m / z: 386.33 [M+H] + .

[0305] Intermediate 38 [ka] The above example was prepared as a mixture of diastereomers in a similar order to intermediate 36 to obtain the desired amino alcohol (42 mg). ESI-MS m / z: 404.09 [M+H] + .

[0306] The compounds in Table 1 were prepared by the same method as in step c (PyBOP or HATU) of Example 1. Most of the compounds were purified by Gilson prep-HPLC, and some by automated column chromatography (silica gel). Many of the aryl acid coupling partners were prepared using intermediates 1-38 or similar procedures. In specific cases, the aryl acid coupling partners were commercially available or prepared by previously described methods. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20

[0307] Intermediate 39

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change

[0308] Intermediate 39 Step b [ka] In a 40 mL vial, the compound from step a (3.3 g, 12 mmol), NaOAc (5 g, 60 mmol), hydroxylamine hydrochloride (4.2 g, 60 mmol), and MeOH (20 mL) were added at room temperature. The resulting mixture was stirred overnight at 60°C under a nitrogen atmosphere. The reaction was monitored by LC-MS. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (5:1) to obtain the desired compound (2.3 g, 66%) as a white solid. ESI-MS m / z: 288.95 [M+H] + .

[0309] Intermediate 39 steps c and d [ka] The compound from step b (1.1 g, 3.8 mmol) and DME (10 mL) were added to a 20 mL vial at 0°C. TFAA (0.9 g, 4 mmol) was added dropwise to the mixture. The resulting mixture was stirred at room temperature for a further 3 hours. The reaction was monitored by TLC. The residue was purified by reverse-phase flash chromatography using MeCN in water to obtain the desired compound (600 mg, 58%) as a white solid. ESI-MS m / z: 270.95 [M+H] + .

[0310] The compound from step c (600 mg, 2.2 mmol), ferrous chloride (56 mg, 0.44 mmol), and DME (10 mL) were added to a 20 mL vial at room temperature. The resulting mixture was stirred at 80 °C for 2 hours under a nitrogen atmosphere. The reaction was monitored by TLC. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (5:1) to obtain the desired compound (400 mg, 67%) as a white solid. ESI-MS m / z: 270.95 [M+H] + .

[0311] Intermediate 39 Step e [ka] In a 50 mL round-bottom flask, the compound from step d (400 mg, 1.5 mmol), cyclopropanol (257 mg, 4.4 mmol), NaH (71 mg, 3 mmol), and DMF (10 mL) were added at 0°C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction product was quenched by adding saturated NH4Cl (aqueous solution). The aqueous layer was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (5:1) to obtain the desired compound (360 mg, 83%) as a white solid. ESI-MS m / z: 293.00 [M+H] + .

[0312] Intermediate 39 Step f [ka] To a solution of the compound from step e (360 mg, 1.2 mmol) in EtOH (4 mL) and DMF (4 mL), Pd(OAc)2 (83 mg, 0.4 mmol), DPPP (304 mg, 0.74 mmol), and TEA (1 mL) were added in a pressure tank. The mixture was pressurized with carbon monoxide to 10 atm at 100°C overnight. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The residue was purified by reverse-phase flash chromatography to obtain the desired compound (300 mg, 85%) as a white solid. ESI-MS m / z: 287.10 [M+H] + .

[0313] Intermediate 39 Step g [ka] In a 50 mL round-bottom flask, the compound from step f (298 mg, 1 mmol), LiOH (125 mg, 5.2 mmol), MeOH (5 mL), and H2O (1 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. The reaction was monitored by LC-MS. The mixture was acidified to pH 6 with HCl (aqueous solution). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired compound (245 mg, 91%) as a grayish-white solid. ESI-MS m / z: 259.10 [M+H] + .

[0314] Intermediate 40 [ka] Intermediate 40 Steps a and b [ka] A mixture of methyl 4-amino-3-methoxybenzoate (50.00 g, 275.95 mmol) and acrolein (30.94 g, 551.87 mmol) in HCl (100 mL) and AcOH (100 mL) was stirred at 100°C for 1 hour under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum and used directly in the next step. ESI-MS m / z: 204.10 [M+H] + .

[0315] The mixture of compounds from step a in DMF, ethyl iodide (76.76 g, 492.16 mmol) and Cs2CO3 (240.52 g, 738.20 mmol), was stirred overnight at room temperature. The resulting mixture was filtered, and the filtrate was washed with ethyl acetate (3 × 300 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with hexane / EA (1:1) to obtain the crude product (25 g) as a yellow solid. ESI-MS m / z: 232.05 [M+H] + .

[0316] Intermediate 40 Step c [ka] A mixture of ethyl 8-methoxyquinoline-6-carboxylate (14.00 g, 60.54 mmol) and NIS (54.48 g, 242.16 mmol) in AcOH (300 mL) was stirred at 100°C for 16 hours under a nitrogen atmosphere. The aqueous layer was extracted with EA. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography and reverse-phase flash chromatography to obtain the desired product (5.5 g, 14%) as a yellow solid. ESI-MS m / z: 358.00 [M+H] + .

[0317] Intermediate 40 Step d [ka] The mixture of the compound from step c (4.30 g, 12.04 mmol) and (1,10-phenanthroline)(trifluoromethyl)copper(I) (5.65 g, 18.06 mmol) in DMF (70 mL) was stirred at 110°C for 4 hours under a nitrogen atmosphere. The mixture was cooled to room temperature. The resulting mixture was poured into water and extracted with EA. The combined organic layer was washed with brine and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (4:1) to obtain the desired product (2.6 g, 72%) as a pale yellow solid. ESI-MS m / z: 300.15 [M+H] + .

[0318] Intermediate 40 Step e [ka] The mixture of the compound from step d (2.60 g, 8.69 mmol) and LiOH (2.08 g, 86.85 mmol) in MeOH (30 mL) and H2O (10 mL) was stirred at room temperature for 2 hours. The mixture was acidified to pH 4 with HCl (2 M aqueous solution). The precipitated solid was collected by filtration and washed with water to obtain the desired product (2.29 g, 97%) as a white solid. ESI-MS m / z: 271.95 [M+H] + .

[0319] Intermediate 41 [ka] The following compounds were prepared in a manner similar to that of intermediate 39. The residue was purified by reverse-phase flash chromatography to obtain the desired product (40 mg, 89%) as a pale yellow solid. ESI-MS m / z: 247.10 [M+H] + .

[0320] Intermediate 42 [ka] The following compounds were prepared in a manner similar to that of intermediate 10. The crude residue was purified by reverse-phase flash chromatography to obtain the desired product (130 mg, 72%) as a white solid. ESI-MS m / z: 247.10 [M+H] + .

[0321] Intermediate 43 [ka] Intermediate 43 Steps a and b [ka] Methyl 3-methoxyquinoline-6-carboxylate (400 mg, 1.841 mmol) was added to a 40 mL vial equipped with a stirring bar, and the material was dissolved in DCM. mCPBA (883 mg, 3.68 mmol) was added, and the reaction mixture was stirred overnight for 20 hours. The reaction mixture was diluted with DCM and quenched with sodium thiosulfate. DCM / MeOH extraction was performed using a phase separator cartridge, and the organic matter was concentrated. The residue was purified by automated column chromatography (silica gel, 0-20% methanol in DCM) to obtain the desired compound (361 mg, 84%). ESI-MS m / z: 234.08 [M+H] + .

[0322] Step a (143 mg, 0.613 mmol) was carried out in 50 mL of rbf containing a stirring bar, and the solid was dissolved in DCM. POCl3 (114 μl, 1.226 mmol) was added, a condenser was attached to the flask, and the reaction mixture was heated at 45°C for 2 hours. The reaction mixture was diluted with DCM and quenched with water and saturated sodium bicarbonate. DCM and DCM / MeOH extraction were performed using a phase separator cartridge, and organic matter was concentrated. The residue was purified by automated column chromatography (silica gel, hexane with 0-90% ethyl acetate) to obtain the desired compound (102 mg, 66%). ESI-MS m / z: 252.04 [M+H] + .

[0323] Intermediate 43 Step c [ka] Pd(PPh3)4 (4.59 mg, 3.97 μmol), step b (40 mg, 0.159 mmol), potassium carbonate (65.9 mg, 0.477 mmol), and methylboronic acid (47.6 mg, 0.795 mmol) were added to a 20 mL vial equipped with a stirring bar. The vial was purged with N2 for 10 minutes. 1,4-dioxane (1.060 ml) was added, and the reaction mixture was heated at 100°C for 18 hours. The reaction mixture was diluted with HCl and quenched with water. HCl and DCM / MeOH extraction were performed using a phase separator cartridge, and organic matter was concentrated. The residue was purified by automated column chromatography (silica gel, hexane, 0-80% HCl) to obtain the desired compound (36 mg, 98%). ESI-MS m / z: 232.10 [M+H] + .

[0324] Intermediate 43 Step d [ka] Step c (36 mg, 0.156 mmol) was added to a 20 mL vial equipped with a stirring bar, and the material was dissolved in THF:MeOH:water (1:1:1, 0.2 M). Lithium hydroxide hydrate (32.7 mg, 0.778 mmol) was added, and the reaction mixture was heated to 45°C and monitored by LC-MS. The vial was cooled to 0°C and acidified to pH=4 with 2 M HCl. No precipitate formed, and the organic matter was concentrated under vacuum. A solid precipitate formed. The mixture was cooled to 0°C and further precipitated. The solid was recovered by vacuum filtration and dried under high vacuum to obtain the desired pdt as a white solid (18 mg, 53%). ESI-MS m / z: 217.82 [M+H] + .

[0325] Example 233 [ka] (S)-7-(4-fluorophenyl)-3-(hydroxymethyl)-5-((S)-1,1,1-trifluoro-2-hydroxy-3-(8-methoxy-2,3-dimethylquinoline-6-carboxamide)propan-2-yl)-2,3-dihydrofluoro[2,3-c]pyridine-3-carboxamide hydrochloride (20 mg, 0.032 mmol) was dissolved in DCM (1 ml), and DAST (1 equivalent, 4.2 μl, 0.032 mmol) was added at 0°C. The reaction mixture was stirred at 0°C for 2 hours. The reaction mixture was quenched by adding MeOH, the solution was concentrated, and purified by HPLC to obtain (S)-3-(chloromethyl)-7-(4-fluorophenyl)-5-((S)-1,1,1-trifluoro-2-hydroxy-3-(8-methoxy-2,3-dimethylquinoline-6-carboxamide)propan-2-yl)-2,3-dihydrofluoro[2,3-c]pyridine-3-carboxamide (3.68 mg, 5.69 μmol, yield 17.88%). ESI-MS m / z: 647.074 [M+H] + .

[0326] Intermediate 44 [ka] The above example was prepared as a mixture of diastereomers in a similar order to intermediate 36 to obtain the desired amino alcohol (320 mg). ESI-MS m / z: 426.04 [M+H] + .

[0327] Intermediate 45 [ka] Intermediate 45 Step a [ka] Methyl 7-bromo-5-iodo-2,3-dihydrofl[2,3-c]pyridine-3-carboxylate (586 mg, 1.526 mmol), tetrahydrofuran (85 mL), and ((chloromethoxy)methyl)benzene (6.94 mL, 50.6 mmol) were added to a 250 mL round-bottom flask. The mixture was cooled to -78 °C, and lithium diisopropylamide (2 M solution in tetrahydrofuran / heptane / ethylbenzene, 9.27 mL, 18.54 mmol) was added dropwise over 15 minutes. The mixture was stirred at -78 °C for 1 hour, and then warmed to 23 °C for 30 minutes. The reaction product was diluted with toluene (100 mL) and water (50 mL). The layers were separated, the aqueous layer was extracted with toluene (3 × 50 mL), and the combined organic layers were dried over Na₂SO₄, filtered, and concentrated. The crude substance was purified by automated column chromatography (silica gel, hexane, 0-10% Â) to obtain the title compound (4.69 g, 55%). ESI-MS m / z: 503.55 / 505.47 [M+H] + .

[0328] Intermediate 45 Step b [ka] In a 100 mL round-bottom flask, the compound from step a (4.69 g, 9.30 mmol) and chloroform (47 mL) were added. The mixture was cooled to 0°C, and methanesulfonic acid was added. The mixture was heated to room temperature for 2 hours with stirring and monitored by LC-MS. The reaction product was diluted with HCl and water. The aqueous layer was extracted, and the combined organic layers were washed with NaHCO3, dried over Na2SO4, filtered, and concentrated. The substance was purified by automated column chromatography (silica gel, hexane with 0-20% HCl) to obtain the title compound (2.25 g, 58%). ESI-MS m / z: 413.46 / 415.42 [M+H] + .

[0329] Intermediate 45 Step c [ka] TIFF0007871278000232.tif1814 In a 50 mL round-bottom flask, the compound from step b (2.12 g, 5.12 mmol), followed by dichloromethane (21 mL), and then 2,6-lutidine (890 μL, 7.68 mmol). The mixture was cooled to -78 °C, and then trifluoromethanesulfonic anhydride (1 M solution in dichloromethane, 5.63 mL, 5.63 mmol) was added dropwise over 10 minutes. The mixture was stirred at -78 °C for 30 minutes and monitored by LC-MS. The mixture was warmed to 0 °C and then diluted with water and dichloromethane. The layers were separated, the aqueous layer was washed with dichloromethane (3 × 25 mL), the combined organic layers were washed with NaHCO3 (saturated aqueous solution) (2 × 10 mL), and then concentrated. The resulting oily substance was dissolved in tetrahydrofuran (2 mL) and cooled to 0°C. Tetrabutylammonium fluoride (1 M, 5.37 mL, 5.37 mmol in tetrahydrofuran) was added to this mixture, and the mixture was heated to room temperature for 19 hours. The reaction product was diluted with HCl and water. The aqueous layer was extracted with HCl (3 × 20 mL), and the combined organic layers were dried over Na₂SO₄ and concentrated. The substance was purified by automated column chromatography (silica gel, hexane, 0-10% HCl) to obtain the title compound (2.00 g, 94%). ESI-MS m / z: 415.47 / 417.40 [M+H] + .

[0330] Intermediate 45 Step d [ka] Ammonia (7M solution in MeOH, 20.6mL, 144 mmol) was added to a 100 mL round-bottom flask containing the compound from step c (2.00 g, 4.81 mmol). The mixture was heated at 45°C for 1.5 hours and monitored by LC-MS. The mixture was cooled to room temperature, concentrated, and the yellow solid was proceeded to the next step without further purification (1.93 g, >99%). ESI-MS m / z: 400.46 / 402.41 [M+H] + .

[0331] Intermediate 45 [ka] The above compounds were prepared as a mixture of diastereomers in a similar order to intermediate 36 to obtain the desired amino alcohol (1.056 g, 85%). ESI-MS m / z: 417.97 [M+H] + .

[0332] Intermediate 46 [ka] Intermediate 46 Step a [ka] In a 250 mL round-bottom flask equipped with a stirring bar, methyl 7-bromo-5-iodo-2,3-dihydrofluoro[2,3-c]pyridine-3-carboxylate (2.044 g, 5.32 mmol) was added, and the material was dissolved in THF (21.29 ml). The flask was cooled to -78 °C, (bromodifluoromethyl)trimethylsilane (1.656 ml, 10.65 mmol) was added, followed by the dropwise addition of lithium diisopropylamide (2.93 ml, 5.86 mmol). The reaction mixture was stirred for 1 hour at -78 °C, and then warmed to room temperature. After 1.5 hours, the reaction mixture was diluted with ethyl acetate and quenched with saturated ammonium chloride. The aqueous layer was extracted with ethyl acetate, the organic matter was dried, filtered, and concentrated. The crude substance was purified by automated column chromatography (silica gel, 0-30% toluene in hexane, large column volume) to obtain the desired product (295.6 mg, 13%). ESI-MS m / z: 433.87 / 435.86 [M+H] + .

[0333] Intermediate 46 Step b [ka] A stirring bar was added to a 40 mL vial containing step a (393.9 mg, 0.908 mmol), and ammonia (3890 μl, 27.2 mmol) was added. The reaction mixture was heated to 45°C and monitored by LC-MS (completed for 1.5 hours). The stirring bar was removed, and the reaction mixture was concentrated. This substance was triturated by DCM to obtain the desired product (360 mg, 96%) as a light brown solid. ESI-MS m / z: 418.87 / 420.87 [M+H] + .

[0334] Intermediate 46 [ka] The following compounds were prepared in the same manner as intermediate 36 to obtain the desired amino alcohol as a mixture of diastereomers (105 mg, 92%). ESI-MS m / z: 436.11 [M+H] + .

[0335] Table 2 below includes examples prepared in a manner similar to step c (PyBOP or HATU) of Example 1. Amine coupling partners were prepared according to intermediates 44–46 or by similar procedures as a mixture of diastereomers. Most compounds were purified by Gilson preparative HPLC. Where indicated as a single diastereomer, the diastereomer was separated by Gilson preparative HPLC. Aryl acid coupling partners were prepared according to intermediates 1–46 or by similar procedures with slight modifications, and also by the procedure found in U.S. Patent Application No. 16 / 930,622. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7]

[0336] Table 3 below includes examples prepared in a manner similar to step c (PyBOP or HATU) of Example 1. Most compounds were purified by Gilson prep-HPLC, and some by automated column chromatography (silica gel). Aryl acid coupling partners were prepared according to intermediates 1-43, or by similar procedures with slight modifications, and also according to the procedure found in U.S. Patent Application No. 16 / 930,622. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0337] Table 4 below includes examples prepared in a similar manner to step c (PyBOP or HATU) of Example 1. Most compounds were purified by Gilson prep-HPLC, and some by automated column chromatography (silica gel). Aryl acid coupling partners were commercially available. [Table 4-1] [Table 4-2] [Table 4-3] Table 4-4 Table 4-5 Table 4-6 Table 4-7 Table 4-8 Table 4-9 Table 4-10 Table 4-11 Table 4-12 Table 4-13 Table 4-14

[0338] Intermediate 47

change

change

[0339] Intermediate 47 Step b [ka] A mixture of the compound from step a (1.5 g, 4.64 mmol), 4-fluorophenylboronic acid (0.97 g, 6.96 mmol), Pd(PPh3)2Cl2 (0.33 g, 0.46 mmol), and Na2CO3 (1.23 g, 11.60 mmol) in THF (10 mL) and H2O (2.5 mL) was stirred at 70°C for 1 hour under a nitrogen atmosphere. The reaction was monitored by TLC. The resulting mixture was filtered, and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution with 33% ethyl acetate in hexane) to obtain the desired product (1.2 g, 76%) as a yellow oil. ESI-MS m / z: 339.05 [M+H] + .

[0340] Intermediate 47 Step c [ka] The solution of the compound from step b (1.2 g, 3.54 mmol) in t-BuOH (40 mL) and H2O (40 mL) was cooled to 0°C. Methanesulfonamide (0.34 g, 3.54 mmol) and AD-MIX-β (8.29 g, 10.64 mmol) were added, and the reaction was stirred overnight at room temperature and monitored by LC-MS. The resulting mixture was poured into water and extracted with ethyl acetate. The combined organic layer was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (MeCN in water, 0.1% FA) to obtain the desired product (1.1 g, 83%) as a grayish-white solid. ESI-MS m / z: 373.25 [M+H] + .

[0341] Intermediate 47 Step d [ka] The solution of the compound from step c (1.7 g, 4.56 mmol) in DCM (70 mL) was cooled to 0°C. TsCl (1.31 g, 6.84 mmol), TEA (1.39 g, 13.69 mmol), and DMAP (0.22 g, 1.82 mmol) were added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by LC-MS. The mixture was acidified to pH 4 with HCl (2 M aqueous solution). The resulting mixture was extracted with CH2Cl2. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution with 66% ethyl acetate in hexane) to obtain the desired product (1.6 g, 66%) as a yellow solid. ESI-MS m / z: 527.30 [M+H] + .

[0342] Intermediate 47 Step e [ka] To a 250 mL round-bottom flask containing step d (1.6 g, 3.04 mmol), NH3 (68 mL, 156 equivalents, 7N in MeOH) was added at room temperature. The resulting mixture was stirred at room temperature for 20 hours and monitored by LC-MS. The solvent was removed, and the crude mixture was dissolved in RINKAN. The organic matter was washed three times with saturated sodium bicarbonate, and the organic matter was concentrated. The crude substance was triturated with DCM to obtain the desired product (590.9 mg, 52%) as a grayish-white solid. ESI-MS m / z: 372.15 [M+H] + .

[0343] Intermediate 48 [ka] Intermediate 48 Step a [ka] In a vial, the compound from intermediate step 36b (6.5 g, 17 mmol) and 1-fluoro-N-methoxy-N-methylcyclopropane-1-carboxamide (3 g, 20.39 mmol) were dissolved in THF (121 ml) and cooled to 0°C. Isopropyl magnesium chloride (16.99 ml, 34.0 mmol) was slowly added, and the reaction mixture was stirred at 0°C. After stirring for 3 hours, water was added at 0°C, and the reaction mixture was warmed to room temperature. The aqueous layer was washed with ethyl acetate, and the combined organic layers were dried over MgSO4 and concentrated. The crude reaction mixture was purified by silica gel column chromatography eluted with 0-50% ethyl acetate / hexane to obtain the title compound (2.8 g, 48%) as a foaming white solid. ESI-MS m / z: 343.17 [M+H] + .

[0344] Intermediate 48 Step b [ka] In a vial, the compound from step a (1.4 g, 4.08 mmol), (4-fluorophenyl)boronic acid (0.685 g, 4.90 mmol), PdCl2(dppf)DCM (0.200 g, 0.245 mmol), and K2CO3 (1.692 g, 12.24 mmol) were dissolved in 1,4-dioxane (16.32 ml) and water (4.08 ml). N2 was sprinkled over the reaction mixture and sealed. The reaction mixture was heated to 90°C. After 4 hours, the reaction mixture was cooled to room temperature and water was added. The aqueous layer was washed with phenylethylamine. The combined organic layers were washed with water and brine, dried over MgSO4, and concentrated. The crude reaction mixture was purified by silica gel column chromatography eluting with 0-70% phenylethylamine / hexane to obtain the title compound (1.12 g, 77%). ESI-MS m / z: 359.06 [M+H] + .

[0345] Intermediate 48 Step c [ka] Methyltriphenylphosphonium bromide (3.00 g, 8.41 mmol) and THF (18.7 ml) were placed in a vial and cooled to 0°C in an ice bath. Potassium tert-butoxide (0.906 g, 8.07 mmol) was slowly added as a solution in THF (2 mL), and the resulting yellow suspension was stirred at 0°C for 30 minutes. The compound from step b (1.205 g, 3.36 mmol) was added as a solution in THF (15 ml). The reaction mixture was stirred at 0°C for 2 hours, then warmed to room temperature and stirred for another 2 hours. The reaction was quenched by adding MeOH (5 mL) followed by water (20 mL). The solution was diluted with ELISA, and the layers were separated. The aqueous layer was washed with ELISA, the combined organic layers were washed with brine, dried over MgSO4, and concentrated. The crude reaction mixture was purified by silica gel column chromatography eluting with 0-80% toluene / hexane to obtain the title compound (741 mg, 61.8%). ESI-MS m / z: 357.39 [M+H] + .

[0346] Intermediate 48 Step d [ka] The above compound was prepared in a manner similar to step e of intermediate 36. The reaction mixture was purified by silica gel column chromatography eluting with 0-80% dimethyl acetone / hexide to obtain the title compound (480 mg, 59%). ESI-MS m / z: 391.33 [M+H] + .

[0347] Intermediate 48 Step e [ka] The above compound was prepared in a manner similar to step f of intermediate 36. After aqueous post-treatment, the title compound (585 mg, 87%) was proceeded to without further purification. ESI-MS m / z: 545.38 [M+H] + .

[0348] Intermediate 48 Step f [ka] The above compound was prepared in a manner similar to step g of intermediate 36. After aqueous post-treatment, the title compound (381 mg, 91%) was proceeded to without further purification. ESI-MS m / z: 390.35 [M+H] + .

[0349] Intermediate 49 [ka] The above compound was prepared in a manner similar to that of intermediate 47 to obtain the desired amino alcohol (473 mg, 64%). ESI-MS m / z: 434.40 [M+H] + .

[0350] Intermediate 50 [ka] The above compound was prepared in a manner similar to that of intermediate 36 to obtain the desired amino alcohol (493 mg, 92%). ESI-MS m / z: 452.25 [M+H] + .

[0351] Intermediate 51 [ka] The above compound was prepared in a manner similar to that of intermediate 47 to obtain the desired amino alcohol (246 mg, 83%). ESI-MS m / z: 424.31 [M+H] + .

[0352] Intermediate 52 [ka] The above compound was prepared in a manner similar to that of intermediate 47 to obtain the desired amino alcohol (147 mg, 95%). ESI-MS m / z: 422.36 [M+H] + .

[0353] Intermediate 53 [ka] Intermediate 53 Step a [ka] 2-chloro-5-fluoropyridine-3-ol (10 g, 68 mmol), K2CO3 (19 g, 136 mmol), I2 (19 g, 75 mmol), and H2O (200 mL) were added to a 500 mL round-bottom flask at room temperature. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere and monitored by LC-MS. The reaction product was quenched by adding saturated sodium hyposulfite (aqueous solution) at room temperature and extracted with ethyl acetate. The residue was purified by silica gel column chromatography (elution with 9% ethyl acetate in hexane) to obtain the desired compound (16.7 g, 90%) as a white solid. ESI-MS m / z: 273.90 [M+H] + .

[0354] Intermediate 53 Step b [ka] In a 250 mL vial, the compound from step a (12.4 g, 45 mmol), (2-methyloxyran-2-yl)methyl-4-methylbenzenesulfonate, K2CO3 (13 g, 91 mmol), KI (9 g, 54 mmol), and DMF (50 mL) were added at room temperature. The resulting mixture was stirred overnight at 50°C under a nitrogen atmosphere. The reaction was monitored by LC-MS. The reactants were quenched with water, and the aqueous layer was extracted with ELISA. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flush to obtain the desired compound (9.6 g, 63%) as a white solid. ESI-MS m / z: 343.80 [M+H] + .

[0355] Intermediate 53 Step c [ka] In a 50 mL three-necked round-bottom flask, the compound from step b (1.5 g, 4 mmol), THF (15 mL), and LDA (2.4 mL, 5 mmol) were added at 0°C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. The reaction was monitored by TLC. The reactants were quenched by adding saturated NH4Cl (aqueous solution) at 0°C. The aqueous layer was extracted with ethyl acetate. The residue was purified by reverse flush to obtain the desired compound (430 mg, 29%) as a white solid. ESI-MS m / z: 343.80 [M+H] + .

[0356] Intermediate 53 Step d [ka] In a 100 mL round-bottom flask, the compound from step c (400 mg, 1 mmol), acetone (10 mL), and Jones' reagent (1 mL) were added at 0°C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 hours. The reaction was monitored by LC-MS. The aqueous layer was extracted with ethyl acetate and concentrated under reduced pressure. The crude product was used directly in the next step without further purification. ESI-MS m / z: 357.85 [M+H] + .

[0357] Intermediate 53 Step e [ka] In a 50 mL round-bottom flask, the crude compound from step d, CDI (726 mg, 4 mmol), and THF (5 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. Then, the mixture was added dropwise to NH3.H2O (50 mL) and stirred for 1 hour. The reaction was monitored by TLC. The aqueous layer was extracted with RINKAN, and the residue was purified by reverse flush to obtain the desired compound (218.8 mg, 69%) as a white solid. ESI-MS m / z: 356.90 [M+H] + .

[0358] Intermediate 54 [ka] The following compounds were prepared in a similar sequence to steps a and b of intermediate 48. ESI-MS m / z:336.95 [M+H] + .

[0359] Intermediate 55 [ka] The above compound was prepared in a manner similar to that of intermediate 36. After aqueous post-treatment, the title compound (45 mg, 63%) was isolated as a white solid. ESI-MS m / z: 386.40 [M+H] + .

[0360] Intermediate 56 [ka] The following compounds were prepared in a similar order to steps a and b of intermediate 48. ESI-MS m / z:351.05 [M+H] + .

[0361] Intermediate 57 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude product was purified by automated column chromatography (silica gel, 10% MeOH in DCM) to obtain the desired product (1.80 g, 57%) as a grayish-white solid. ESI-MS m / z: 462.05 [M+H] + .

[0362] Intermediate 58 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude substance was purified by preparative TLC (silica gel, 10% MeOH in DCM containing NH3) to obtain the desired product (508 mg, 74%) as a white solid. ESI-MS m / z: 468.05 [M+H] + .

[0363] Intermediate 59 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude substance was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (450 mg, 72%) as a white solid. ESI-MS m / z: 434.10 [M+H] + .

[0364] Intermediate 60 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude substance was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (468.9 mg, 92%) as a white solid. ESI-MS m / z: 433.95 [M+H] + .

[0365] Intermediate 61 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude product was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product as a white solid. ESI-MS m / z: 434.10 [M+H] + .

[0366] Intermediate 62 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude product was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product as a white solid. ESI-MS m / z: 452.00 [M+H] + .

[0367] Intermediate 63 [ka] The above compound was prepared using the corresponding arylboronic acid pinacol ester in a manner similar to that of intermediate 36. The arylboronic acid pinacol ester was prepared from the corresponding bromide by Pd-catalyzed borylation. The crude product was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (355 mg, 67%) as a white solid. ESI-MS m / z: 452.10 [M+H] + .

[0368] Intermediate 64 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 36. The crude substance was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (570.3 mg, 87%) as a white solid. ESI-MS m / z: 452.10 [M+H] + .

[0369] Intermediate 65 [ka] 3-bromo-7-fluoro-1-benzothiophene (1.29 g, 5.60 mmol), bis(pinacolato)diborone (1.42 g, 5.59 mmol), KOAc (1.65 g, 16.81 mmol), Pd(dppf)Cl2.CH2Cl2 (365 mg, 0.45 mmol), and dioxane (18 mL) were added to a 40 mL sealed tube at room temperature. The resulting mixture was stirred at 110 °C for 1 hour under a nitrogen atmosphere. The reaction was monitored by LC-MS. The mixture was cooled to room temperature. (R)-7-bromo-3-methyl-5-(3,3,3-trifluoropropane-1-en-2-yl)-2,3-dihydrofluoro[2,3-c]pyridine-3-carboxamide (787 mg, 2.24 mmol), K2CO3 (0.93 g, 6.72 mmol), Pd(dppf)Cl2.CH2Cl2 (365 mg, 0.45 mmol), and H2O (2 mL) were added, and the mixture was stirred at 80°C for a further 2 hours. The resulting mixture was filtered, and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (460 mg, 48%) as a brown oil. ESI-MS m / z: 422.95 [M+H] + .

[0370] Intermediate 66 [ka] The above compound was prepared in a manner similar to that of intermediate 36, which has intermediate 65. The crude substance was purified by preparative TLC (silica gel, 8% MeOH in DCM containing NH3) to obtain the desired product (251.3 mg, 73%) as a white solid. ESI-MS m / z: 456.00 [M+H] + .

[0371] Intermediate 67 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (silica gel, 10% MeOH in DCM containing NH3) to obtain the desired product (291.1 mg, 61%) as a white solid. ESI-MS m / z: 440.00 [M+H] + .

[0372] Intermediate 68 [ka] The above compounds were prepared using the corresponding arylboronic acid in a manner similar to that of intermediates 47 and 65. The crude material was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (114.2 mg, 49%) as a white solid. ESI-MS m / z: 428.05 [M+H] + .

[0373] Intermediate 69 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (MeOH in DCM containing silica gel and NH3) to obtain the desired product (220 mg, 64%) as a white solid. ESI-MS m / z: 406.10 [M+H] + .

[0374] Intermediate 70 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (MeOH in DCM containing silica gel and NH3) to obtain the desired product (267.8 mg, 64%) as a white solid. ESI-MS m / z: 406.10 [M+H] + .

[0375] Intermediate 71 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (MeOH in DCM containing silica gel and NH3) to obtain the desired product (262.1 mg, 53%) as a white solid. ESI-MS m / z: 406.15 [M+H] + .

[0376] Intermediate 72 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (silica gel, 6% MeOH in DCM containing NH3) to obtain the desired product (250 mg, 68%) as a white solid. ESI-MS m / z: 424.00 [M+H] + .

[0377] Intermediate 73 Steps a and b [ka] Methyl 3-amino-4-fluorobenzoate (50 mg, 0.296 mmol), isocyanatocyclopropane (24.56 mg, 0.296 mmol), and EtOH (2 mL) were added to a vial. The resulting mixture was stirred at room temperature for 20 hours. After removing the solvent, the residue was purified by automated column chromatography (using a 4 g silica gel column eluting with 0-50% siRNA / hexane) to obtain the desired product (30 mg, 40.2%) as a pale yellow foam. ESI-MS m / z: 253.10 [M+H] + .

[0378] Step a (90 mg, 0.357 mmol) and 2N NaOH (1 mL) were added to a vial containing THF / H2O (2 mL, 9:1) solvent. After stirring overnight at room temperature, the mixture was neutralized to approximately pH 3 by adding 1N HCl. The solid precipitated was filtered and dried in an oven to obtain the desired product (85 mg, 100%). ESI-MS m / z: 239.20 [M+H] + .

[0379] Intermediate 74 [ka] Ethyl 6-hydroxyimidazo[1,2-a]pyridine-2-carboxylate (60 mg, 0.291 mmol), 2-bromopropane (165 μl, 1.164 mmol), and DMF (0.582 ml) were added to a 2 mL MW vial. Cesium carbonate (284 mg, 0.873 mmol) was then added. The reaction vial was capped and heated in a microwave reactor at 140°C for 2 hours. The reaction mixture was diluted with ethyl acetate and water. The organic layer was washed with brine, dried, concentrated, and used in the next step without further purification. ESI-MS m / z: 249.12 [M+H] + .

[0380] Intermediate 75 [ka] Ethyl 3-bromo-2-oxopropanoate (68.5 μl, 0.546 mmol) and 5-(trifluoromethoxy)pyridine-2-amine (81 mg, 0.455 mmol) were dissolved in DME (1.137 ml) and MeOH (1.137 ml). The reaction mixture was heated at 80°C for 18 hours. After removing the solvent, the residue was purified by flash chromatography to obtain the desired product (78 mg, 62.6%). ESI-MS m / z: 275.057 [M+H] + .

[0381] Intermediate 76 [ka] Methyl 2-(1-hydroxycyclopropyl)-7-methoxy-2H-indazole-5-carboxylate (55 mg, 0.210 mmol) was dissolved in dichloromethane (1.049 ml) at 0°C. Thionyl chloride (61.2 μl, 0.839 mmol) was added, and the mixture was stirred at 50°C for 18 hours. The reaction mixture was directly purified by flash chromatography to obtain the desired product (34 mg, 57.8%). ESI-MS m / z: 281.061 [M+H] + .

[0382] Intermediate 77 [ka] Intermediate 77 Step a [ka] 5-bromo-1-methoxy-2-methyl-3-nitrobenzene (1 g, 4 mmol), NBS (0.8 g, 4.5 mmol), AIBN (0.13 g, 0.8 mmol), and CCl4 (20 mL) were added to a 100 mL round-bottom flask at room temperature. The resulting mixture was stirred overnight at 80°C under a nitrogen atmosphere and monitored by LC-MS. The reaction was quenched by adding water, and the resulting mixture was extracted with ethyl acetate. The combined organic matter was dried over sodium sulfate, filtered, and concentrated. The crude product was used directly in the next step without further purification. ESI-MS m / z: 325.90 [M+H] + .

[0383] Intermediate 77 Step b [ka] In a 100 mL round-bottom flask, the compound from step a (1.33 g, 4 mmol), aminocyclopropane (2.34 g, 41 mmol), K2CO3 (1.13 g, 8 mmol), and DMF (10 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 3 hours. The reaction was monitored by LC-MS. The aqueous layer was extracted with phenylethylamine. The residue was purified by silica gel column chromatography (elution with 16% ethyl acetate in hexane) to obtain the desired compound (1.07 g, 87%) as a white solid. ESI-MS m / z: 301.00 [M+H] + .

[0384] Intermediate 77 Step c [ka] In a 100 mL three-necked round-bottom flask, the compound from step b (500 mg, 1.7 mmol) and THF (6 mL) were added at 0°C. NaOH (664 mg, 17 mmol) in H2O (6 mL) was added over 2 minutes, followed by the addition of zinc (326 mg, 5 mmol) in 6 portions over 3 hours, and the mixture was stirred for a further 1 hour. The reaction was monitored by LC-MS. The mixture was acidified to pH 3 with HCl (aqueous solution). The aqueous layer was extracted with CH2Cl2 and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and washed with hexane to obtain the desired compound (310 mg, 70%) as a white solid. ESI-MS m / z: 267.00 [M+H] + .

[0385] Intermediate 77 Step d [ka] In a 30 mL pressure tank reactor, the compound from step c (240 mg, 0.9 mmol), dppp (222 mg, 0.5 mmol), Pd(OAc)2 (60 mg, 0.3 mmol), DMF (4 mL), EtOH (4 mL), and TEA (1 mL) were added at room temperature. The resulting mixture was stirred overnight at 100 °C under a 15 atm CO atmosphere. The reaction was monitored by LC-MS. The resulting mixture was filtered, and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase silica gel chromatography (MeCN / H2O) to obtain the desired compound (210 mg, 90%) as a white solid. ESI-MS m / z: 261.10 [M+H] + .

[0386] Intermediate 77 Step e [ka] In a 50 mL round-bottom flask, the compound from step d (470 mg, 1.8 mmol), lithium hydroxide (216 mg, 9 mmol), methanol (10 mL), and H2O (3 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 1 hour. The reaction was monitored by LC-MS. The mixture was acidified to pH 6 with HCl (aqueous solution). The resulting mixture was washed with water and evaporated to obtain the desired compound (317.8 mg, 76%) as a yellow solid. ESI-MS m / z: 233.10 [M+H] + .

[0387] Intermediate 78 [ka] Intermediate 78 Step a [ka] Methyl 3-[(cyclopropylamino)methyl]-5-methoxy-4-nitrobenzoate (3 g, 11 mmol), KOH (6 g, 107 mmol), MeOH (20 mL), and H2O (2 mL) were added to a 50 mL round-bottom flask at room temperature. The resulting mixture was stirred at 80°C for 72 hours under a nitrogen atmosphere. The reaction was monitored by TLC. The mixture was acidified to pH 6 with HCl (aqueous solution). The residue was purified by reverse-phase silica gel chromatography (MeCN / H2O) to obtain the desired compound (580 mg, 21%) as a white solid. ESI-MS m / z: 263.10 [M+H] + .

[0388] Intermediate 78 Step b [ka] The compound from step a (1.28 g, 5 mmol) and AcOH (6 mL) were added to a 40 mL vial at room temperature. The resulting mixture was stirred at 120 °C for 3 days under a nitrogen atmosphere. The reaction was monitored by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase silica gel chromatography (MeCN / H2O) to obtain the desired compound (0.96 g, 79%) as a brown solid. ESI-MS m / z: 249.05 [M+H] + .

[0389] Intermediate 79 [ka] Intermediate 79 Step a [ka] A solution of 5-bromo-7-methoxy-1H-indazole (3.00 g, 13.21 mmol) in MeCN (50 mL) was treated overnight at room temperature with 1-(benzenesulfonyl)cyclopropan-1-ol (3.14 g, 15.85 mmol) and TEA (1.60 g, 15.85 mmol). The reaction was monitored by LC-MS. The resulting mixture was extracted with ethyl acetate and concentrated. The residue was purified by silica gel column chromatography (eluted with 50% ethyl acetate in hexane) to obtain the desired compound (2.40 g, 64%) as a yellow solid. ESI-MS m / z: 283.00 [M+H] + .

[0390] Intermediate 79 Step b [ka] A solution of the compound obtained from step a (2.43 g, 8.58 mmol) in DCM (20 mL) was treated with BAST (5.69 g, 25.74 mmol) at 0°C to room temperature for 1.5 hours. The reaction was monitored by LC-MS and quenched with water. The aqueous layer was extracted with RINKAN and concentrated. The residue was purified by silica gel column chromatography (eluted with 50% ethyl acetate in hexane) to obtain the desired compound (1.2 g, 49%) as a yellow solid. ESI-MS m / z: 285.00 [M+H] + .

[0391] Intermediate 79 steps c and d [ka] The following compounds were prepared according to the same procedure as in steps d and e of intermediate 77 to obtain the desired product (312.6 mg, 58%) as a white solid. ESI-MS m / z: 250.95 [M+H] + .

[0392] Intermediate 80 [ka] The following compound was prepared according to the same procedure as for intermediate 79 to obtain the desired product (170.3 mg, 76%) as a white solid. ESI-MS m / z: 276.95 [M+H] + .

[0393] Intermediate 81 [ka] Intermediate 81 Steps a and b [ka] A mixture containing 5-bromo-1-fluoro-3-methyl-2-nitrobenzene (5 g, 21.37 mmol), cyclopropanol (2.5 g, 42.73 mmol), and Cs2CO3 (21 g, 64 mmol) in DMF (20 mL) was stirred at 50°C for 2 hours under an N2 atmosphere. The resulting mixture was extracted with siRNA (3 × 200 mL). The combined organic layer was washed with water and concentrated under reduced pressure to obtain the crude product without further purification.

[0394] The solution / mixture of the compound from step a (5 g, 18.38 mmol), Fe (10.3 g, 183.76 mmol), and NH4Cl (9.8 g, 183.76 mmol) in EtOH (40 mL) and H2O (20 mL) was stirred at 80°C for 2 hours. The residue was purified by silica gel column chromatography (ethyl acetate in hexane) to obtain the desired product (3.6 g, 81%) as a yellow solid. ESI-MS m / z: 242.00 [M+H] + .

[0395] Intermediate 81 steps c and d [ka] The compound from step b (1.5 g, 6.2 mmol) was added to a 50% aqueous solution of fluoroboric acid (9.8 mL) at room temperature and stirred for 5 minutes. The mixture was cooled in an ice bath for 10 minutes, and an aqueous solution of NaNO2 (900 mg, 13.04 mmol) in 1.7 mL of H2O was added to the mixture. The reaction mixture was stirred at 10°C for 30 minutes, during which time the product precipitated. The cooled reaction mixture was filtered through a Buchner funnel, the solid product was washed with a small amount of H2O, MeOH, and Et2O, and dried under high vacuum to obtain the crude product as a brown solid. ESI-MS m / z: 253.00 [M+H] + .

[0396] In a dry flask, 18-crown-6 (0.05 g, 0.2 mmol) and potassium acetate (2.9 g, 29.4 mmol) were dried under high vacuum for 1 hour. CHCl3 (70 mL) was added, and the mixture was stirred at room temperature for 10 minutes. Then, step c was gradually added to the mixture under an N2 atmosphere. The reaction mixture was stirred at room temperature for 3 hours, filtered, and the residue was washed with CHCl3. The filtrate was washed with water (3 × 40 mL), the organic layer was dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by flash column chromatography (hexane / RINKAN) to obtain the desired product (1.8 g, 82%) as a yellow solid. ESI-MS m / z: 253.00 [M+H] + .

[0397] Intermediate 81 Steps e, f, g [ka] The following intermediates were prepared in a manner similar to steps a-c of intermediate 6, and the desired product (330 mg, 87%) was obtained as a yellow solid. ESI-MS m / z: 269.10 [M+H] + .

[0398] Intermediate 82 [ka] Intermediate 82 Steps a~d [ka] The following intermediates were prepared using cyclopropaneethanol in a manner similar to steps a-d of intermediate 81, to obtain the desired product (1.85 g, 66%) as a pale yellow solid. ESI-MS m / z: 266.90 [M+H] + .

[0399] Intermediate 82 Steps e~h [ka] The following intermediates were prepared in a manner similar to steps a-d of intermediate 79 to obtain the desired product (298 mg, 60%) as a white solid. ESI-MS m / z: 291.10 [M+H] + .

[0400] Intermediate 83 [ka] The following intermediates were prepared in a manner similar to that of intermediate 6, and the desired product (260 mg, 69%) was obtained as a white solid. ESI-MS m / z:247.00 [M+H] + .

[0401] Intermediate 84 [ka] The following intermediate was prepared in a manner similar to that of intermediate 6 to obtain the desired product. ESI-MS m / z:231.00 [M+H] + .

[0402] Intermediate 85 [ka] Intermediate 85 Step a [ka] A solution / mixture of methyl 6-chloro-5-methoxypyridine-3-carboxylate (1 g, 4.96 mmol), ethinylcyclopropane (0.66 g, 9.92 mmol), Pd(PPh3)4 (1.15 g, 0.99 mmol), CuI (0.47 g, 2.48 mmol), and Et3N (1.51 g, 14.88 mmol) in 1,4-dioxane (30 mL) was stirred overnight at 90°C under an N2 atmosphere. The residue was purified by silica gel column chromatography (elution with 50% ethyl acetate in hexane) to obtain the desired product (1 g, 87%) as a yellow solid. ESI-MS m / z: 232.00 [M+H] + .

[0403] Intermediate 85 Steps b~d [ka] To a solution of the compound from step a (1 g, 4.32 mmol) in CH2Cl2 (100 mL), a solution of amino 2,4,6-trimethylbenzenesulfonate (1.21 g, 5.62 mmol) in CH2Cl2 (50 mL) was added under ice cooling, and the reaction mixture was stirred for a further 1 hour. Et2O (12 mL) was added to the reaction mixture to precipitate the crystals. The filtrate was filtered off and then dried under reduced pressure to obtain the crude product as a pale yellow solid. ESI-MS m / z: 247.00 [M+H] + .

[0404] The solution / mixture of the compound from step b (500 mg, 2.02 mmol) and K2CO3 (559 mg, 4.04 mmol) in MeOH (50 mL) was stirred at room temperature under an N2 atmosphere for 2 hours. The residue was purified by silica gel column chromatography (elution with ethyl acetate in hexane) to obtain the desired product (220 mg, 44%) as a yellow solid. ESI-MS m / z: 247.00 [M+H] + .

[0405] The compound from step c (220 mg, 0.89 mmol) and LiOH (214 mg, 8.93 mmol) were mixed in MeOH (10 mL) and H2O (10 mL) and stirred at room temperature for 2 hours. The mixture was acidified with HCl to pH 5-6. The resulting mixture was filtered, and the filtered cake was collected to obtain the desired product (160 mg, 77%) as a white solid. ESI-MS m / z: 233.00 [M+H] + .

[0406] Intermediate 86 [ka] Intermediate 86 Step a [ka] A mixture of 3-methoxy-6-bromopyridine 2-ylamine (1.0 g, 3.6 mmol) and chloroacetaldehyde (1.2 mL, 50% wt in water) in EtOH (12 mL) was stirred under reflux for 1 hour and then concentrated under reduced pressure. The residue was partitioned between a saturated solution of sodium bicarbonate and ethyl acetate. The organic layer was separated, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography (Si-PPC, MeOH / Et2O, gradient 0:100~1:99) to obtain the desired product as a beige solid (960 mg, 88%). ESI-MS m / z: 227.00 [M+H] + .

[0407] Intermediate 86 steps b and c [ka] The following example was prepared in a manner similar to that of intermediate 77, and the desired product (340 mg, 49%) was obtained as a yellow solid. ESI-MS m / z: 192.95 [M+H] + .

[0408] Intermediate 87 [ka] Intermediate 87 Steps a and b [ka] To a solution of 2,6-dibromo-4-methoxypyridine (5 g, 18.73 mmol), tert-butylcarbamate (2.19 g, 18.73 mmol), and Cs2CO3 (12.21 g, 37.46 mmol) in 1,4-dioxane (60 mL), Pd(OAc)2 (0.42 g, 1.87 mmol) and xanthophos (1.08 g, 1.87 mmol) were added. The reaction mixture was heated at 100 °C for 16 hours under an N2 atmosphere. After cooling the reaction mixture to room temperature, water was added and the mixture was extracted with EA (50 mL x 3 times). The combined organic layer was washed with water and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (2 g, 35%) as a yellow solid. ESI-MS m / z:303.00 [M+H] + .

[0409] The compound (1.5 g) from step a and TFA (8 mL) in DCM (20 mL) were stirred at room temperature for 2 hours. The residue product was purified by reverse-phase flushing under the following conditions (water (0.05% FA) / MeOH) to obtain the desired product (520 mg) as a white solid. ESI-MS m / z: 203.00 [M+H] + .

[0410] Intermediate 87 Steps c~e [ka] The following example was prepared in a manner similar to that of intermediate 86 to obtain the desired product (300 mg, 86%) as a yellow solid. ESI-MS m / z: 192.95 [M+H] + .

[0411] Intermediate 88 [ka] Intermediate 88 Step a [ka] A solution of 6-bromo-3-methoxypyridine-2-amine (1 g, 4.93 mmol) and 2-bromo-1-cyclopropylethanone (963 mg, 5.91 mmol) in EtOH (10 mL) was stirred overnight at 80°C. The reaction mixture was quenched with water, and the resulting mixture was extracted with EA (3 × 200 mL). The combined organic layer was washed with NaHCO3 (3 × 200 mL) and dried over anhydrous NaSO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (360 mg, 27%) as a yellow solid. ESI-MS m / z: 267.00 [M+H] + .

[0412] Intermediate 88 steps b and c [ka] The following example was prepared in a manner similar to that of intermediate 86 to obtain the desired product (110 mg, 62%) as a yellow solid. ESI-MS m / z:233.00 [M+H] + .

[0413] Intermediate 89 [ka] The following example was prepared in a manner similar to that of intermediate 86 to obtain the desired product. ESI-MS m / z:206.95 [M+H] + .

[0414] Intermediate 90 [ka] Intermediate 90 Step a [ka] A solution / mixture of 3-cyclopropoxy-2-nitropyridine (1.8 g, 1 mmol), Fe (5.58 g, 100 mmol), and NH4Cl (5.34 g, 100 mmol) in EtOH (60 mL) and H2O (30 mL) was stirred at 80°C for 2 hours. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (1.4 g, 93%) as a yellow solid. ESI-MS m / z: 151.00 [M+H] + .

[0415] Intermediate 90 Step b [ka] The solution / mixture of the compound from step a (1.4 g, 9.32 mmol) and Br2 (2.98 g, 18.64 mmol) in AcOH (30 mL) was stirred overnight at room temperature. The resulting mixture was quenched with saturated sodium thiosulfate and extracted with EA. The combined organic layer was washed with NaHCO3 solution (3 × 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to obtain the desired product (1.4 g, 66%) as a yellow solid. ESI-MS m / z: 229.00 [M+H] + .

[0416] Intermediate 90 Steps c~e [ka] The following example was prepared in a manner similar to that of intermediate 86 to obtain the desired product. ESI-MS m / z:219.00 [M+H] + .

[0417] Intermediate 91 [ka] Intermediate 91 Steps a and b [ka] A solution of 3-hydroxy-4-aminobenzoic acid (10 g, 65.30 mmol) and methacrolein (9.15 g, 130.60 mmol) in HCl (40 mL) and AcOH (60 mL) was stirred at 100°C for 1 hour under an N2 atmosphere. The combined layers were concentrated under reduced pressure to obtain the desired product (5.6 g, 42%) as a brown solid. ESI-MS m / z: 204.00 [M+H] + .

[0418] The compound from step a (5.6 g, 27.56 mmol) and the solution / mixture in MeOH (50 mL) and H2SO4 (5 mL) were stirred under reflux at 80°C for 1 hour. The mixture was neutralized to pH 7 with NaOH solution. The resulting mixture was extracted with EA (3 × 300 ml). The combined organic layer was concentrated under reduced pressure to obtain the desired product (2.5 g, 42%) as a yellow solid. ESI-MS m / z: 218.00 [M+H] + .

[0419] Intermediate 91 steps c and d [ka] The solution of the compound from step b (500 mg, 2.30 mmol), 2-bromo-1,1-difluoroethane (667 mg, 4.6 mmol), and K2CO3 (954 mg, 6.91 mmol) in DMF (20 mL) was stirred overnight at 80°C. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (500 mg, 62%) as a yellow solid. ESI-MS m / z: 282.00 [M+H] + .

[0420] The compound from step c (500 mg, 1.78 mmol) and LiOH (426 mg, 17.78 mmol) were mixed in MeOH (20 mL) and H2O (20 mL) and stirred at room temperature for 30 minutes. The mixture was acidified to pH 7 with HCl. The crude product was recrystallized from water to obtain the desired product (270 mg, 57%) as a yellow solid. ESI-MS m / z: 267.95 [M+H] + .

[0421] Intermediate 92 Steps a and b [ka] A solution of methyl 8-hydroxy-3-methylquinoline-6-carboxylate (3 g, 13.81 mmol), sodium 2-chloro-2,2-difluoroacetate (3.16 g, 20.72 mmol), and Cs2CO3 (9 g, 27.61 mmol) in DMF (30 mL) was stirred at 80°C for 4 hours under an N2 atmosphere. The resulting mixture was extracted with EA (3 × 200 ml). The combined organic layer was washed with water (3 × 200 ml) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (2 g, 54%) as a yellow solid. ESI-MS m / z: 268.00 [M+H] + .

[0422] The compound from step a (3 g, 11.23 mmol) and LiOH (2.7 g, 112.26 mmol) in MeOH (30 mL) and H2O (30 mL) were stirred at room temperature for 4 hours. The mixture / residue was acidified to pH 7 with HCl, the resulting mixture was filtered, and the filter cake was washed with water. The residue was purified by reverse-phase flash chromatography under the following conditions (silica gel, MeCN in water, 10% to 50% gradient over 10 minutes) to obtain the desired product (800 mg, 28%) as a yellow solid. ESI-MS m / z: 254.20 [M+H] + .

[0423] Intermediate 93 Steps a and b [ka] A mixture of methyl 4-amino-3-chlorobenzoate (1.5 g, 8.08 mmol) and (2Z)-2-chlorobuta-2-enal (1.27 g, 12.12 mmol) in HCl (10 mL) and AcOH (15 mL) was stirred at 100°C for 20 minutes. The reaction was monitored by LC-MS. The mixture was cooled to room temperature and concentrated under vacuum. The residue was dissolved in MeOH (20 mL) and H2SO4 (2 mL) and stirred at 80°C for a further 1 hour. The resulting mixture was poured into water and extracted with EA. The combined organic layer was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (110 mg, 5%) as a yellow solid. ESI-MS m / z: 269.90 [M+H] + .

[0424] In a 100 mL round-bottom flask, the compound from step a (110 mg, 0.40 mmol), THF (4 mL), MeOH (1 mL), LiOH (97 mg, 4.07 mmol), and H2O (1 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 3 hours. The mixture was acidified to pH 5 with HCl (1 M aqueous solution). The product precipitated. The precipitated solid was collected by filtration and washed with water to obtain the desired product (65 mg). ESI-MS m / z: 256.00 [M+H] + .

[0425] Intermediate 94 [ka] Intermediate 94 Step a [ka] A solution of methyl 3-bromo-5-methoxy-4-nitrobenzoate (3.60 g, 12.43 mmol) in toluene (20 mL) was treated with tributyl (1-ethoxyethenyl) stannane (5.39 g, 14.91 mmol) and Pd(dppf)Cl2 (1.82 g, 2.49 mmol), and the mixture was stirred at 110°C for 2 hours under a nitrogen atmosphere. The reaction was monitored by LC-MS. The resulting mixture was filtered, and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution with 25% ethyl acetate in hexane) to obtain the compound (2.69 g, 77%) as a yellow oil. ESI-MS m / z: 282.20 [M+H] + .

[0426] Intermediate 94 Step b [ka] A solution of the compound (2.69 g, 9.56 mmol) from step a in DCM (30 mL) was treated with HCl (3 mL) overnight at room temperature. The reaction was monitored by LC-MS. The aqueous layer was extracted with CH2Cl2. The residue was purified by silica gel column chromatography (elution with 50% ethyl acetate in hexane) to obtain the compound (1.82 g, 75%) as a white solid. ESI-MS m / z: 253.85 [M+H] + .

[0427] Intermediate 94 steps c and d [ka] In a 250 mL round-bottom flask, the compound from step b (1.82 g, 7.18 mmol) was added. The solution of NaOAc (21.83 g, 266.09 mmol) in MeOH (90 mL) and THF (90 mL) was treated with SnCl2 (18.19 g, 94.92 mmol). The mixture was stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction was monitored by LC-MS. The resulting mixture was filtered, and the filtrate was washed with ethyl acetate. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (7:3) to obtain the compound (851 mg, 54%) as a white solid. ESI-MS m / z: 222.10 [M+H] + .

[0428] In a 250 mL round-bottom flask, the compound from step c (840 mg, 3.80 mmol) and THF (40 mL) were added at room temperature. A solution of LiOH (909 mg, 37.97 mmol) in MeOH (10 mL) and H2O (10 mL) was added, and the mixture was stirred at room temperature for 5 hours. The reaction was monitored by LC-MS. The mixture was acidified to pH 4 with HCl, and the product was precipitated. The precipitated solid was collected by filtration and washed with H2O to obtain the compound (646 mg, 81%) as an orange solid. ESI-MS m / z: 207.95 [M+H] + .

[0429] Intermediate 95 [ka] The following compound was prepared using a procedure similar to that for intermediate 94, and the desired product (741 mg, 88%) was obtained as an orange solid. ESI-MS m / z: 178.10 [M+H] + .

[0430] Table 5 below includes examples prepared in a manner similar to step c (PyBOP or HATU) of Example 1. Most compounds were purified by preparative HPLC (ACN / H2O, 20–90%, 25 min), and some by automated column chromatography (silica gel). Aryl acid coupling partners were prepared according to intermediates 1–95, or by similar procedures with slight modifications, and also according to the procedure found in U.S. Patent Application No. 16 / 930,622. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7] [Table 5-8] [Table 5-9] [Table 5-10] [Table 5-11] [Table 5-12] [Table 5-13] [Table 5-14] Table 5-15 Table 5-16 Table 5-17 Table 5-18 Table 5-19 Table 5-20 Table 5-21 Table 5-22 Table 5-23 Table 5-24 Table 5-25 Table 5-26 Table 5-27 Table 5-28 Table 5-29 Table 5-30 Table 5-31 [Table 5-32] [Table 5-33] [Table 5-34]

[0431] Intermediate 96 [ka] Intermediate 96 Step a [ka] 5-bromoisatin (1 g, 4 mmol) and DCM (5 mL) were added to a 100 mL round-bottom flask at room temperature. DAST (1.43 g, 9 mmol) was added dropwise. The resulting mixture was stirred at 0°C for 1 hour under a nitrogen atmosphere. The reaction was monitored by LC-MS. The reaction product was quenched with water, and the aqueous layer was extracted with EA. The residue was purified by silica gel column chromatography (eluted with 25% ethyl acetate in hexane) to obtain the desired compound (900 mg, 82%) as a pale yellow solid.

[0432] Intermediate 96 Step b [ka] A solution of the compound from step a (300 mg, 1.2 mmol), bis(pinacolato)diborone (461 mg, 1.8 mmol), Pd(dppf)Cl2CH2Cl2 (197 mg, 0.24 mmol), and KOAc (356 mg, 4 mmol) in a dioxane (5 mL) mixture was stirred at 90°C for 2 hours under a nitrogen atmosphere. The reaction was monitored by TLC. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flush to obtain the desired compound (297 mg, 83%) as a yellow solid.

[0433] Intermediate 97 [ka] The following compounds were prepared in a manner similar to step b of intermediate 96 above to obtain the desired product, which was then used directly as the crude product in the next step.

[0434] Intermediate 98 [ka] In a 50 mL round-bottom flask, n-BuLi (2.8 mL, 5.6 mmol) was added dropwise to a solution of 1-bromo-2-chloro-3,4-difluorobenzene (850 mg, 3.7 mmol) in THF (30 mL) under an N2 atmosphere at -78 °C. The reaction mixture was stirred at -78 °C for 5 minutes. Then, B(OMe)3 (583 mg, 5.6 mmol) was added dropwise, and the mixture was stirred further at room temperature for 30 minutes. The mixture was acidified to pH 5 with HCl (3 M aqueous solution). The reaction product was quenched with saturated NH4Cl, and then the mixture was extracted with RINKAN. The combined organic extract was washed with brine, dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the desired crude compound (790 mg, 109%) as a yellow oil. The crude product was used directly in the next step without further purification.

[0435] Example 890 [ka] Example 890 Process a [ka] To a 250 mL round-bottom flask containing intermediate 36 step c (5.09 g, 14.50 mmol), THF (24.16 mL), acetone (24.16 mL), and water (24.16 mL) were added. The flask was cooled to 0°C, NMO (4.25 g, 36.2 mmol) was added, followed by potassium osminate dihydrate (0.230 g, 0.623 mmol). The reaction mixture was stirred for 10 minutes, then warmed to room temperature and stirred overnight for 20 hours. Sodium sulfite was added, the mixture was diluted with water, and the reaction mixture was stirred for 20 minutes. The mixture was diluted with ELISA, and the aqueous layer was extracted. The combined organic matter was dried over sodium sulfate, filtered, and concentrated. The crude mixture was purified by automated column chromatography (silica gel, hexane, 0-100% ethyl acetate) to obtain the desired product as a white solid (4.12 g, 74%) and a mixture of diastereomers. ESI-MS m / z:385.21 [M+H] + .

[0436] Example 890 Process b [ka] In a 250 mL round-bottom flask containing step a (4.118 g, 10.69 mmol), DCM (36 mL, 0.3 M) was added. DMAP (0.065 g, 0.535 mmol) was added, followed by TEA (4.47 ml, 32.1 mmol). The flask was cooled to 0°C, TsCl (2.242 g, 11.76 mmol) was added, and the mixture was stirred for 10 minutes. The mixture was then warmed to room temperature and monitored by LC-MS (1 hour). The reaction product was concentrated, and the crude residue was purified by automated column chromatography (silica gel, hexane, 0-100% ethyl acetate) to obtain the desired product as a white solid (3.92 g, 99%). ESI-MS m / z: 367.19 [M+H] + .

[0437] Example 890 Process c [ka] In a 500 mL round-bottom flask containing step b (3.92 g, 10.70 mmol), ammonia (239 mL, 1670 mmol, 7N in MeOH) was added at 0°C. The reaction mixture was stirred for 10 minutes, warmed to room temperature, and monitored by LC-MS (3.5 hours). The stirred material was then removed, and the reaction mixture was concentrated. The crude product was dissolved in ELISA, and the organic matter was washed three times with saturated sodium bicarbonate. The organic matter was concentrated, and the crude product was triturated with DCM / hexane to obtain a white solid (2.41 g, 59%) as a mixture of diastereomers. ESI-MS m / z: 384.21 [M+H] + .

[0438] Example 890 Process d [ka] In a 100 mL round-bottom flask equipped with a stirring bar, step c (1.000 g, 2.60 mmol) and 2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (0.605 g, 2.60 mmol) were added. The solid was dissolved in N,N-dimethylformamide (13.02 mL, 0.2 M), and DIPEA (0.909 mL, 5.21 mmol) was added. The vial was cooled to 0°C, and PyBOP (1.626 g, 3.12 mmol) was added. The reaction mixture was stirred for 10 minutes, warmed to room temperature, and monitored by LC-MS (1 hour). The reaction mixture was quenched with saturated ammonium chloride and diluted with ethyl acetate. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were dried over sodium sulfate, filtered, and concentrated. The crude mixture was purified by automated column chromatography (silica gel, hexane, 0-100% ethyl acetate) to obtain the desired product as an oil. The oily substance was dissolved in phenylethylamine, washed with water and brine to remove DMF, and concentrated to obtain a mixture of diastereomers as a white solid (597 mg, 38%). ESI-MS m / z: 598.12 [M+H] + .

[0439] Example 891 [ka] PdCl2 (dppf) (6.11 mg, 8.36 μmol), potassium carbonate (26.0 mg, 0.188 mmol), (4-cyclopropylphenyl)boronic acid (16.24 mg, 0.100 mmol), and Example 890 (50 mg, 0.084 mmol) were added to two drum vials equipped with a stirring bar and septum caps. The materials were dissolved in 1,4-dioxane (0.3 mL, 4:1, 0.2 M) and water (0.084 mL) and purged with nitrogen. The reaction mixture was heated to 90°C and monitored by LC-MS (2 hours). The reaction mixture was diluted with siRNA, filtered through a short pad of silica gel, washed with siRNA, and the organic matter was concentrated. The crude material was purified, and the diastereomers were separated by preparative HPLC (ACN / H2O, 20-90%, 25 min) to obtain the desired product as a white solid (3 mg, 6%). ESI-MS m / z:636.26 [M+H] + .

[0440] Example 892 [ka] The following example was purified as a white solid (3 mg, 6%) by preparative HPLC (ACN / H2O, 20-90%, 25 min) as described in Example 891. ESI-MS m / z: 636.26 [M+H] + .

[0441] Table 6 below includes examples prepared using Example 890 in a similar manner to Example 891. More palladium and boronic acid were added when conversion needed to be accelerated. Most of the compounds were purified, and the diastereomers were separated by preparative HPLC (ACN / H2O, 20–90%, 25 min). If a mixture of diastereomers was reported, it is likely they could not be separated by HPLC. [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6] [Table 6-7] [Table 6-8] [Table 6-9]

[0442] Table 7 below includes examples prepared in a manner similar to step c (PyBOP or HATU) of Example 1. Most compounds were purified by preparative HPLC (ACN / H2O, 20–90%, 25 min). Where indicated as a single diastereomer, they were separated during preparative HPLC purification. Amine coupling partners were prepared in a manner similar to intermediates 45 and 46. Aryl acid coupling partners were prepared according to intermediates 1–95, or by similar procedures with slight modifications, and also by the procedure found in U.S. Patent Application No. 16 / 930,622. [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6]

[0443] Intermediate 99 [ka] Intermediate 99 Steps a and b [ka] 5-(benzyloxy)-2-(hydroxymethyl)-1H-pyridine-4-one (5 g, 21.62 mmol), Trt-Cl (6.63 g, 23.78 mmol), and DMAP (2.91 g, 23.78 mmol) were suspended in DMF (50 mL) and stirred overnight at 95°C. The reaction was monitored by LC-MS. After cooling to room temperature, the mixture was treated with cold water and stirred for 30 minutes. The precipitated solid was collected by filtration, washed with water and MeOH, and dried to obtain the desired product (8.8 g, 86%) as a pale gray solid. ESI-MS m / z: 474.30 [M+H] + .

[0444] To a solution of the compound from step a (4.4 g, 9.29 mmol) in THF (120 mL), H2O (12 mL), and NaOH 2M aqueous solution (6 mL), Pd / C (2.2 g, 20.67 mmol) was added under a nitrogen atmosphere. The resulting mixture was stirred under a hydrogen atmosphere at room temperature for 2 hours. The reaction was monitored by TLC. The resulting mixture was filtered, and the filtrate was washed with acetonitrile. The filtrate was concentrated under reduced pressure. To a suspension of the residue in DCM (100 mL), DMAP (2.27 g, 18.58 mmol) and thiophosgene (1.58 g, 13.75 mmol) were added. After stirring at room temperature for 1 hour, the reaction was monitored by TLC. The reactants were quenched with water and extracted with CH2Cl2. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with P / EA (10:1) to obtain the desired product (3.3 g, 83%) as a yellow solid. ESI-MS m / z: 448.20 [M+Na] + .

[0445] Intermediate 99 steps c and d [ka] To a solution of the compound from step b (3.3 g, 7.75 mmol) in DCM (100 mL), HF pyridine (33 mL) and DBDMH (3.77 g, 13.18 mmol) were added, and the temperature was maintained below -60°C. The reaction mixture was heated to 0°C over 20 minutes. After stirring at that temperature for 2 hours, the reaction mixture was quenched with 2 M NaOH aqueous solution and extracted with CH2Cl2. The combined organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to obtain the desired product (740 mg, 50%) as a yellow oil. ESI-MS m / z: 190.05 [M+H] + .

[0446] Intermediate 99 Step e [ka] To a solution of the compound from step d (550 mg, 2.94 mmol) in acetone (20 mL) and H2O (10 mL), 2-methyl-2-butene (2.06 g, 29.40 mmol), NaH2PO4 (529 mg, 4.41 mmol), and NaClO2 (532 mg, 5.88 mmol) were added. The resulting mixture was stirred at room temperature for 2 hours. The mixture was treated with NaHSO3 (1.07 g, 10.28 mmol), concentrated under reduced pressure, and the acetone was removed. The mixture was treated with brine (10 mL) and extracted twice with EA / THF (1:1). The combined organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired product (151.8 mg, 25%) as a white solid. ESI-MS m / z: 203.90 [M+H] + .

[0447] Intermediate 100 [ka] Intermediate 100 steps and b [ka] A mixture of 5-bromo-7-methoxy-1H-indazole (500 mg, 2.2 mmol), Cs2CO3 (2.15 g, 6.6 mmol), and CBr2F2 (5 mL) in ACN (25 mL) was stirred overnight at room temperature under an N2 atmosphere. The residue was purified by reverse-phase flash chromatography to obtain the desired product (200 mg, 26%) as a yellow oil. ESI-MS m / z: 355.00 [M+H] + .

[0448] The solution of the compound from step a (400 mg, 1.12 mmol) and AgBF4 (656 mg, 3.37 mmol) in DCM (20 mL) was stirred at room temperature for 2 hours. The resulting mixture was washed with 3 × 100 mL of NaHCO3. The resulting mixture was extracted with EA (3 × 100 mL). The combined organic layer was washed with water (3 × 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA to obtain the desired product (300 mg, 90%) as a yellow solid. ESI-MS m / z: 295.00 [M+H] + .

[0449] Intermediate 100 steps c and d [ka] The following compounds were prepared according to the same procedure as in steps d and e of intermediate 77 to obtain the desired product (200 mg, 72%) as a white solid. ESI-MS m / z: 275.00 [M+H] + .

[0450] Intermediate 101 [ka] Intermediate 101 Steps a, b and c [ka] A mixture of 2-amino-5-bromo-3-methoxybenzaldehyde (9.6 g, 41.73 mmol) and urea (37.59 g, 625.92 mmol) was stirred at 180°C for 2 hours. The mixture was cooled to room temperature and poured into ice water. The precipitated solid was collected by filtration, washed with water, and dried under vacuum to obtain the crude product (11 g) as a gray solid. ESI-MS m / z: 254.85 [M+H] + .

[0451] A solution of the compound from step a (11 g, 43.12 mmol) in phosphorus oxychloride (100 mL) was stirred at 110°C for 5 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in EA and poured into ice water with vigorous stirring. The resulting mixture was extracted with EA. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (2:1) to obtain the desired product (2.4 g, 20%) as a pale yellow solid. ESI-MS m / z: 272.95 [M+H] + .

[0452] The mixture of the compound from step b (2.4 g, 8.77 mmol) and NaOMe (0.47 g, 8.77 mmol) in MeOH (30 mL) was stirred at room temperature for 1 hour, and then heated under reflux for 1 hour. The reaction was monitored by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was stirred with water. The precipitated solid was collected by filtration, washed with water, and dried under vacuum to obtain the desired product (2.2 g, 93%) as a pale yellow solid. ESI-MS m / z: 268.85 [M+H] + .

[0453] Intermediate 101 Steps d and e [ka] The following compounds were prepared according to the same procedure as in steps d and e of intermediate 77 to obtain the desired product (658.6 mg, 77%) as a white solid. ESI-MS m / z: 235.00 [M+H] + .

[0454] Intermediate 102 [ka] Intermediate 102 Steps a and b [ka] To a solution of methyl 4-amino-3-methoxybenzoate (4 g, 22.07 mmol) in HCl (15 mL), methacrolein (3.87 g, 55.19 mmol) was added. The resulting mixture was stirred at 100 °C for 5 hours. The reaction was monitored by LC-MS. The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. This yielded 8-methoxy-3-methylquinoline-6-carboxylic acid (4 g, 83%) as a brown crude solid. ESI-MS m / z: 218.05 [M+H] + .

[0455] To a solution of the compound (4 g, 18.41 mmol) from step a in MeOH (120 mL), SOCl2 (8.76 g, 73.65 mmol) was added. The resulting mixture was stirred at 80°C for 40 minutes. The reaction was monitored by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in water (40 mL). The mixture was basicized to pH 7 with 1 M NaHCO3 aqueous solution, extracted, and evaporated. The residue was purified by silica gel column chromatography eluting with PE / EA (1:2) to obtain the compound (1.8 g, 42%) as a brown solid. ESI-MS m / z: 232.00 [M+H] + .

[0456] Intermediate 102 steps c and d [ka] To a solution of the compound from step b (1.8 g, 7.78 mmol) in DCM (20 mL), m-CPBA (4.03 g, 23.35 mmol) was added in fractions at 0°C. The resulting mixture was stirred overnight at room temperature. The reaction was monitored by LC-MS. The resulting mixture was washed with 1 M NaOH. The aqueous layer was basicized to pH 2 with concentrated HCl. The resulting mixture was filtered, and the filtrate was washed with water. The filtrate was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. ESI-MS m / z: 233.95 [M+H] + .

[0457] A solution of the compound (5 g crude) from step c and POCl3 (42.73 g, 278.707 mmol, 13 equivalents) was stirred at 95°C for 1 hour. The reaction mixture was slowly added to water (20 mL), and the reaction was monitored by LC-MS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flush to obtain the title compound (100 mg, 1.85%) as a white solid. ESI-MS m / z: 251.95 [M+H] + .

[0458] Intermediate 103 [ka] This intermediate was prepared in a manner similar to that of intermediate 102. (80 mg, 70%) ESI-MS m / z: 277.95 [M+H] + .

[0459] Intermediate 104 [ka] The following compounds were prepared in a manner similar to that of intermediate 102 (64 mg, 74%). ESI-MS m / z: 318.10 [M+H] + .

[0460] Intermediate 105 [ka] The above compound was prepared in a manner similar to that of intermediate 102 (32 mg, 46%). ESI-MS m / z: 268.15 [M+H] + .

[0461] Intermediate 106 [ka] Intermediate 106 Steps a, b and c [ka] 2,5-Dichloro-3-fluoropyridine (10 g, 60.25 mmol), (4-methoxyphenyl)methanol (9.16 g, 66.27 mmol), Cs2CO3 (39.26 g, 121 mmol), and DMF (40 mL) were added to a 250 mL round-bottom flask at room temperature. The resulting mixture was stirred at 80 °C for 1 hour. The reaction was monitored by LC-MS. The aqueous layer was extracted with ELISA. The resulting mixture was washed with brine. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography to obtain the desired compound (14 g, 81%) as a white solid. ESI-MS m / z: 284.25 [M+H] + .

[0462] The compound from step a (5 g, 18 mmol) and DCM (24 mL) were added to a 100 mL round-bottom flask at room temperature. CF3COOH (6 g, 52.62 mmol) was added dropwise to the mixture. The resulting mixture was stirred overnight at room temperature. The reaction was monitored by LC-MS. The aqueous layer was extracted with CH2Cl2. The resulting mixture was concentrated under vacuum, and the crude product was used directly in the next step without further purification. ESI-MS m / z: 165.10 [M+H] + .

[0463] In a 100 mL round-bottom flask, the compound from step c (6.2 g, 37.81 mmol), I2 (10.56 g, 41.59 mmol), K2CO3 (10.45 g, 75.61 mmol), and H2O (40 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LC-MS. The reactants were quenched with saturated sodium hypochlorite (aqueous solution) at 0°C. The mixture was acidified to pH 6 with concentrated HCl. The aqueous layer was extracted with ethyl acetate. The residue was purified by silica gel column chromatography to obtain the desired compound as a white solid. ESI-MS m / z: 290.15 [M+H] + .

[0464] Intermediate 106 d and e [ka] In a 250 mL round-bottom flask, the compound from step c (3.9 g, 13.45 mmol), (2-methyloxyran-2-yl)methyl-4-methylbenzenesulfonate (3.91 g, 16.14 mmol), KI (2.2 g, 13.45 mmol), K2CO3 (3.7 g, 26.91 mmol), and DMF (20 mL) were added at room temperature. The resulting mixture was stirred overnight at 50 °C. The reaction was monitored by LC-MS. The reactants were quenched with saturated NH4Cl (aqueous solution) at 0 °C. The aqueous layer was extracted with ELISA. The residue was purified by reverse-phase flash chromatography to obtain the desired compound (3.0 g, 65%) as a yellow oil. ESI-MS m / z: 359.95 [M+H] + .

[0465] The compound from step d (1.85 g, 5.13 mmol) and LDA (2 M in THF) (3.9 mL) were added to a 50 mL three-necked round-bottom flask at 0°C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. The reaction was monitored by LC-MS. The desired product was detected by LC-MS. The reaction product was quenched with saturated NH4Cl (aqueous solution), extracted with EA, and the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to obtain the desired compound as a yellow oil. ESI-MS m / z: 360.15 [M+H] + .

[0466] Intermediate 106 Steps f and g [ka] In a 250 mL round-bottom flask, the compound from step e (6.6 g, 18.33 mmol), acetone (20 mL), and Jones' reagent (8 mL, 40.39 mmol) were added in portions at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 hours. The reaction was monitored by LC-MS. The aqueous layer was extracted with ethyl acetate. The resulting liquid was dried under vacuum. The crude product was used directly in the next step without further purification. ESI-MS m / z: 374.20 [M+H] + .

[0467] In a 250 mL round-bottom flask, the compound from step f (3.5 g, 9.35 mmol), CDI, and THF (50 mL) were added at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. Then, the mixture was added dropwise to NH3H2O ​​(400 mL) and stirred for 2 hours. The reaction was monitored by LC-MS. The aqueous layer was extracted with ethyl acetate. The residue was purified by reverse-phase flash chromatography to obtain the desired compound (1 g, 28.65%) as a white solid. ESI-MS m / z: 372.90 [M+H] + .

[0468] Intermediate 107 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. (95 mg, 91%) ESI-MS m / z: 470.16 [M+H] + .

[0469] Intermediate 108 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. (476 mg, 86%) ESI-MS m / z: 452.13 [M+H] + .

[0470] Intermediate 109 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. (382 mg, 68%) ESI-MS m / z: 470 / 10 [M+H] + .

[0471] Intermediate 110 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (MeOH in DCM containing silica gel and NH3) to obtain the desired product (305 mg, 59%) as a white solid. ESI-MS m / z: 434.15 [M+H] + .

[0472] Intermediate 111 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was purified by preparative TLC (silica gel, MeOH in DCM containing NH3) to obtain the desired product (170 mg, 72%) as a white solid. ESI-MS m / z: 406.16 [M+H] + .

[0473] Intermediate 112 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was concentrated to dryness, redissolved in toluene, and washed with 5 × 1 mL of saturated aqueous solution of NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the desired product (1.311 g, 80%) as a white powder. ESI-MS m / z: 436.23 [M+H] + .

[0474] Intermediate 113 [ka] The above compound was prepared using the corresponding arylboronic acid in a manner similar to that of intermediate 47. The crude substance was concentrated to dryness, redissolved in siRNA, and washed with 5 × 1 mL of saturated aqueous solution of NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the desired product (589.7 mg, 82%) as a yellow powder. ESI-MS m / z: 408.29 [M+H] + .

[0475] Table 8 below includes examples prepared in a manner similar to step c (PyBOP or HATU) of Example 1. Most compounds were purified by preparative HPLC (ACN / H2O, 20–90%, 25 min), and some by automated column chromatography (silica gel). Aryl acids and amine coupling partners were prepared according to intermediates 1–113, or by similar procedures with slight modifications, and also according to the procedure found in U.S. Patent Application No. 16 / 930,622. Table 8-1 Table 8-2 Table 8-3 Table 8-4 Table 8-5 Table 8-6 Table 8-7 Table 8-8 Table 8-9 Table 8-10 Table 8-11 Table 8-12 Table 8-13

[0476] Intermediate 114

change

[0477] Intermediate 115 [ka] The following intermediates were prepared in a manner similar to that of Example 890, and the desired product was obtained as a pale yellow solid (1.63 g, 62%) as a mixture of diastereomers. ESI-MS m / z: 572.28 [M+H] + .

[0478] Table 9 below includes examples prepared in a similar manner to Example 891 using Example 890 and intermediate 115. More palladium and boronic acid were added when it was necessary to accelerate conversion. Most compounds were purified, and the diastereomers were separated by preparative HPLC (ACN / H2O, 20–90%, 25 min). If a mixture of diastereomers was reported, it is likely that they could not be separated by HPLC. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5]

[0479] Example 1268 [ka] Example 1228 (15 mg, 0.024 mmol) was dissolved in EtOH (0.5 mL) and MeOH (0.5 mL), and palladium carbon (10% by weight, 0.8 mg, 0.00075 mmol) was added. The air in the headspace of the reaction vessel was replaced with H2 from a balloon, and the mixture was stirred under H2 for 24 hours. The reaction mixture was then filtered through a short Celite pad, and the filtrate was concentrated to obtain the product. (14 mg, 93%) ESI-MS m / z: 638.29 [M+H] + .

[0480] Example 1269 [ka] In a vial, Example 890 (30 mg, 0.05 mmol) was dissolved in DMF (1 mL). Morpholine (44 mg, 0.5 mmol) and K2CO3 (69 mg, 0.5 mmol) were added, the vial was sealed, and the mixture was heated at 100°C for 24 hours. The reaction mixture was cooled to room temperature and diluted with H2O (1 mL). The aqueous layer was washed with ethyl acetate, and the combined organic layers were dried over MgSO4 and concentrated under reduced pressure. The crude product was purified by preparative HPLC (ACN / H2O, 20-90%, 25 min) to obtain the title compound (5 mg, 16%). ESI-MS m / z: 605.27 [M+H] + .

[0481] Example 1270 [ka] The following example was prepared in a format similar to Example 1269. ESI-MS m / z:639.20 [M+H] + .

[0482] Example 1271 [ka] The following example was prepared in a format similar to Example 1269. ESI-MS m / z:639.20 [M+H] + .

[0483] Table 10 below includes examples prepared using Example 890 and intermediate 115 in a manner similar to step c (PyBOP or HATU) of Example 1 and Example 891. Most compounds were purified by preparative HPLC (ACN / H2O, 20–90%, 25 min), and some by automated column chromatography (silica gel). Aryl acids and amine coupling partners were prepared according to intermediates 1–113, or by similar procedures with slight modifications, and also according to the procedure found in U.S. Patent Application No. 16 / 930,622. [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5] [Table 10-6]

[0484] Prepare the following examples in Table 11 using the same procedure as described above. [Table 11-1] [Table 11-2] [Table 11-3] [Table 11-4] [Table 11-5] [Table 11-6] [Table 11-7] [Table 11-8] [Table 11-9] [Table 11-10] [Table 11-11] [Table 11-12] [Table 11-13] [Table 11-14]

[0485] Assay Method for RSV-A assay Hep-2 cells (originally derived from tumors that grew in irradiated, cortisone-treated, weaned rats injected with epidermoid carcinoma tissue from the larynx of a 56-year-old male, but later found to be indistinguishable from HeLa cells by PCR DNA analysis) were used to culture genotype A, "long" strain RSV. RSV was inoculated into flasks, and the virus stock was collected when the cytopathic effect (CPE) exceeded 90%. The virus stock in 25% sucrose medium was rapidly frozen using liquid nitrogen to enhance viral stability. The virus stock titer was measured using 3-fold virus dilutions across 8,000 cells / well and 96-well plates to achieve a tissue culture infectious dose of 50% (TCID). 50 The titer was quantified by ) and cultured for 4 days. The viral stock titer was also quantified by the plaque-forming unit assay, as described elsewhere.

[0486] After extensive parameter testing, the final assay is performed as follows: Hep-2 cells are seeded in 60 inner wells of a 96-well plate at 8,000 cells / well in 50 μL of growth medium (phenol red-free DMEM, 1% L-Glut, 1% Penn / Strep, 1% non-essential amino acids, 10% thermo-inactivated FBS). Twofold serial dilutions of the control and test compounds are added to the wells in double series in total volumes of 25 μL. Then, the virus stock is added to the wells in 25 μL volumes at an infection multiplicity (MOI) of 0.1, bringing the total volume of each well to 100 μL. MOI is expressed as PFU / mL, or TCID if PFU / mL is unavailable. 50The calculation is performed using the following method. Each 96-well plate has six control columns containing cells and viruses but no compounds (negative control, maximum CPE), six columns containing cells but no compounds or viruses (positive control, minimum CPE), and six columns containing neither cells, viruses, nor compounds (background plate / reagent control). The control wells containing cells but no viruses are given an additional 25 μL of growth medium containing the same amount of sucrose as the wells receiving the virus stock to maintain constant medium and volume conditions. The outer wells of the plate are filled with 125 μL of moat media (DMEM, 1% Penn / Strep) to act as a thermal and evaporation barrier (moat) around the test wells. After a 5-day incubation period, the plates are read using ATPlite (50 μL added per well) to quantify the amount of ATP present in each well (a measure of cell health). The assay plates are read using an Envision luminometer. These data are used to calculate the EC of each compound. 50 Calculate (Table 12). EC 50 The range is as follows: A < 0.2 μM; B > 0.2 μM. [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4] [Table 12-5] [Table 12-6] [Table 12-7] [Table 12-8] [Table 12-9] [Table 12-10] [Table 12-11] [Table 12-12] [Table 12-13] [Table 12-14]

[0487] Method for HMPV antiviral assay Method-A: HMPV antiviral activity was evaluated using a recombinant version of HMPV CAN97-83 engineered to contain a green fluorescent protein (eGFP) coding sequence enhanced at the 3' end of the viral genome (MPV-GFP1, ViraTree). One day prior to the assay, Vero E6 cells (ATCC#CCL-7) were seeded in 96-well cell plates at a density of 12,000 cells / 100 μL / well. On the day of screening, cell culture medium was aspirated from the wells, and cells were washed twice with serum-free Eagle Modified Essential Medium (EMEM, ATCC#) (SF-EMEM) containing 1% penicillin-streptomycin (Invitrogen). Cell washing was performed by dispensing 100 μL of SF-EMEM per well and immediately aspirating the washing medium from the wells. Following a second washing cycle, serum-free OptiMEM (Invitrogen, catalog number) (SF-OptiMEM) containing 0.5 μg / mL TPCK-trypsin (VENDOR) and 1% penicillin-streptomycin was added to cells at a rate of 50 μL / well. The compounds were added to 96-well plates using a JANUS automated liquid processing system (VENDOR). The compounds were first diluted 1:50 in an intermediate 96-well plate containing SF-OptiMEM, and then transferred to assay plates (25 μL / well). Each test compound was tested in 2-well sets at a total of 8 points using 1 / 2 serial dilutions, starting at final concentrations of 8 μM or 2 μM. Viral infection was performed by preparing a working stock of MPV-GFP1 with an infection multiplicity (MOI) equal to 0.05 / 25 μL and ali-coating 25 μL of viral inoculum into the compound and positive control wells. SF-OptiMEM was added to the appropriate wells (25 μL / well) to serve as a virus-free negative control for the assay. The final DMSO concentration in all wells was 0.5%. The plates were incubated at 32°C and 5% CO2 for 5 days.

[0488] After a 5-day incubation period, eGFP fluorescence intensity was measured at (X)nM wavelength using a Spectramax i3X plate reader (VENDOR). The viral inhibition percentage was calculated using the following formula: y=[100-(X Q / X P )] × 100 (In the formula, X Q This is the fluorescence intensity measured in wells containing recombinant MPV-GFP1 infected compound-treated cells, and X P (This is the average fluorescence intensity measured in wells containing untreated cells infected with recombinant virus.) Then, EC 50 The values ​​were calculated by nonlinear regression using a four-parameter curve rheological equation. The curve-fitted model used was the XLFit Dose-Response One Site Model 200: y=(A+(B / (1+((x / C)^D)))) (In the equation, A is the minimum y value, B is the maximum y value, and C is logEC) 50 (These are values, and D is the gradient coefficient). Using these data, the EC of each compound is calculated. 50 Calculate (Table 13). EC 50 The range is as follows: A < 0.5 μM; B > 0.5 μM. [Table 13-1] [Table 13-2] [Table 13-3]

[0489] Method-B: In vitro HMPV antiviral activity was evaluated using clinical isolates TN / 1501 / A1 and LLC-MK2 cells (ATCC#CCL-7), which are immortalized renal epithelial cell lines derived from rhesus monkeys (Macaca mulatta).

[0490] The compound was resuspended in 10 mM dimethyl sulfoxide (DMSO) and added to a 384-well source plate. The compound was diluted and transferred to a 384-well assay plate using an Echo-650 automated liquid processing system (Beckman Coulter, Indiana). The test compound was evaluated in two consecutive dilutions at the highest concentration of 10 μM, followed by three-fold serial dilutions to obtain a total of 10 concentration points. A DMSO control well was also included in the assay plate, representing either infection or non-infection, and served as both a positive and negative control.

[0491] TN / 1501 / A1 virus infection was performed by suspension with LLC-MK2 cells. The cells were washed twice with PBS and removed from the cell culture flask containing 0.25% trypsin-EDTA (Thermo Fisher Scientific, Massachusetts). The trypsin-EDTA was inactivated by resuspending it in OptiMEM (Thermo Fisher Scientific, Massachusetts) containing 2% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The cells were pelleted by centrifugation at 800 rpm for 5 minutes, the supernatant was removed, and the cells were suspended in PBS + 100 μg / mL CaCl2. This process was repeated twice. The cells were then resuspended in serum-free (SF)-OptiMEM containing 4 μg / mL TPCK-trypsin (Sigma Aldrich, Missouri), 1% penicillin-streptomycin (Thermo Fisher Scientific, Massachusetts), and 100 μg / mL CaCl2. The cells were counted and seeded at a density of 5,000 cells / well and 12.5 μL / well.

[0492] Viral infection was performed by adding 12.5 μL per well to achieve a multiple of infection (MOI) of 0.014. SF-OptiMEM (12.5 μL / well) was added to the appropriate wells to serve as a virus-free negative control for the assay. The final concentration of TPCK-trypsin was 2 μg / mL. The plates were incubated at 37°C and 5% CO2 for 6 days.

[0493] After 6 days of incubation, 12.5 μL of ATP-Lite (Perkin Elmer, Massachusetts) was added to each well, and the raw luminescence values ​​were determined using Envision 2104 (Perkin Elmer, Massachusetts). The average raw luminescence values ​​of the positive control wells containing only cells and viruses were subtracted from all conditions tested, and the percentage of healthy cells was determined by dividing these values ​​by the average of the negative control wells containing only cells. Subsequently, EC 50 The values ​​were calculated by nonlinear regression using a four-parameter curve rheological equation. The curve-fitted model used was the XLFit Dose-Response One Site Model 200: y = (A + (B / (1 + ((x / C)^D)))) (where A is the minimum y value, B is the maximum y value, and C is log EC) 50 (These are values, and D is the gradient coefficient). Using these data, the EC of each compound is calculated. 50 Calculate (Table 14). EC 50 The range is as follows: A < 0.5 μM; B > 0.5 μM. [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4] [Table 14-5] [Table 14-6] [Table 14-7] [Table 14-8] [Table 14-9]

[0494] While the present invention has been specifically illustrated and described with reference to its preferred embodiments, it will be understood by those skilled in the art that various modifications can be made to the form and details without departing from the scope of the invention as encompassed by the appended claims.

Claims

1. A compound selected from the compounds listed below, or a pharmaceutically acceptable salt thereof. Table 1-1 Table 1-2 Table 1-3

2. The aforementioned compound 【Chemistry 1】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

3. The aforementioned compound 【Chemistry 2】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

4. The aforementioned compound 【Transformation 3】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

5. The aforementioned compound 【Chemistry 4】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

6. The aforementioned compound 【Transformation 5】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

7. The aforementioned compound 【Transformation 6】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

8. The aforementioned compound 【Transformation 7】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

9. The aforementioned compound 【Transformation 8】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

10. The aforementioned compound 【Chemistry 9】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

11. The aforementioned compound 【Chemistry 10】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

12. The aforementioned compound 【Chemistry 11】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

13. The aforementioned compound 【Chemistry 12】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

15. The pharmaceutical composition according to claim 14 for the treatment or prevention of respiratory syncytial virus infection.

16. The pharmaceutical composition according to claim 14 for the treatment or prevention of human metapneumovirus infection.

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