Spiro pyrrolidine derived antiviral drugs

Abstract

The present invention relates to novel spiro pyrrolidine derived antiviral agents. Disclosed is a compound of Formula (Ia): and pharmaceutically acceptable salts thereof, which have inhibitory activity against coronavirus replication. The present invention further relates to pharmaceutical compositions comprising a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof, and to methods of treating or preventing a coronavirus infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof.

Description

[0001] The inventors include: Guojun Wang, Shengui Chen, Yung He, Ma Jun, Xing Xuechao, Cao Hui, Gao Xurizi, Peng Xiaowen, Long Jiang, Li Wei, Zhang Jiajun, J. D. Panarose, N. T. Kent, S. Bartlett, and Ke Rixin.

[0002] Related Applications

[0003] This application claims priority to U.S. Provisional Patent Application No. 63 / 117,170, filed November 23, 2020, U.S. Provisional Patent Application No. 63 / 142,663, filed January 28, 2021, U.S. Application No. 17 / 479,244, filed September 20, 2021, and U.S. Provisional Patent Application No. 17 / 479,530, filed September 20, 2021. The entire teachings of the above applications are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0004] The present invention relates to compounds and methods of inhibiting coronavirus replicative activity by targeting 3C-like protease (sometimes referred to as “3CLpro,” “main protease,” or “Mpro”) with a therapeutically effective amount of a 3C-like protease inhibitor. The present invention also relates to pharmaceutical compositions comprising a coronavirus 3C-like protease inhibitor, and administering an effective amount of such a coronavirus 3C-like protease inhibitor to a mammal. BACKGROUND

[0005] Coronaviruses are a family of single-stranded, positive-sense RNA viruses with a viral envelope, classified in the order Nidovirales. The coronavirus family includes pathogens of many animal species, including humans, horses, cows, pigs, birds, cats, and monkeys, and has been well known for over 60 years. For example, isolation of the mouse coronavirus prototype strain JHM was reported in 1949. Coronaviruses are a common virus that usually causes mild to moderate upper respiratory illness in humans and are named for the crown-like projections on the surface of their envelope. There are four major subgroups of coronaviruses, called alpha, beta, gamma, and delta coronaviruses, and the first coronaviruses were discovered in the mid-1960s. Coronaviruses known to infect humans include alpha coronaviruses 229E and NL63; and beta coronaviruses OC43, HKU1, SARS-coronavirus (the coronavirus that causes severe acute respiratory syndrome or SARS), and MERS-coronavirus (the coronavirus that causes Middle East respiratory syndrome or MERS). People commonly become infected with human coronaviruses 229E, NL63, 0C43, and HKU1, and symptoms usually include mild to moderate upper respiratory illness of short duration, such as runny nose, cough, sore throat, and fever. Occasionally, human coronaviruses can cause lower respiratory illness, such as pneumonia, although this is more common in people with heart-and-lung disease or compromised immune systems or in the elderly. The transmission of common human coronaviruses is not completely understood. However, human coronaviruses are likely spread from infected people to others through the air by coughing and sneezing, and by close personal contact, such as touching or shaking hands. These viruses can also spread by touching contaminated objects or surfaces, and then touching the mouth, nose, or eyes.

[0006] Coronaviruses are enveloped, positive-sense, single-stranded RNA viruses. The coronavirus genomic RNA has a 5' cap structure and a 3' poly-A tail and contains at least six open reading frames (ORFs). The first ORF (ORF1A / b) is directly translated into two polyproteins: pp1a and pp1ab. These polyproteins are processed into 16 non-structural proteins by a 3C-like protease (3CLpro, also known as main protease (Mpro). These non-structural proteins are involved in the production of subgenomic RNAs that encode four structural proteins, namely envelope, membrane, spike, and nucleocapsid proteins, and other accessory proteins. Thus, the 3C-like protease is understood to play a critical role in the coronavirus life cycle.

[0007] 3CLpro is a cysteine protease involved in most cleavage events within the precursor polyprotein. Active 3CLpro is a homodimer containing two protomers with a Cys-His dyad between domains I and II. 3CLpro is conserved among coronaviruses and 3CLpro substrates of different coronaviruses share some common features. Since there is no human homolog of 3CLpro, it is an ideal antiviral target. Although there are reports that certain compounds can inhibit 3CLpro activity, they have not been approved as coronavirus therapies. (See WO 2004101742 A2, US 2005 / 0143320 Al, US 2006 / 0014821 Al, US 2009 / 0137818 Al, WO 2013 / 049382 A2, WO 2013 / 166319 Al, WO2018042343, WO2018023054, WO2005113580, and WO2006061714).

[0008] Due to this highly unmet clinical need, there is a need for more effective treatments for coronavirus infection. The present invention provides compounds that inhibit the life cycle of coronaviruses and methods of making and using these compounds. These compounds can be used to treat or prevent coronavirus infection and reduce the occurrence of disease complications such as organ failure or death. SUMMARY

[0009] The present invention relates to novel antiviral compounds, pharmaceutical compositions comprising the compounds, and methods of using the compounds to treat or prevent viral, especially coronavirus, infection in a subject in need of such treatment. The compounds of the present invention inhibit a protein encoded by a coronavirus or interfere with the life cycle of a coronavirus and can also be used as antiviral agents. In addition, the present invention provides methods of making the compounds.

[0010] In certain embodiments, the present invention provides compounds of Formula (Ia) and pharmaceutically acceptable salts, esters, and prodrugs thereof,

[0011]

[0012] wherein:

[0013] A is selected from:

[0014] 1) -R 11 ;

[0015] 2) -OR 12 ; and

[0016] 3) -NR 13 R 14 ;

[0017] B is optionally substituted aryl or optionally substituted heteroaryl;

[0018] X is selected from:

[0019] 1) -CN;

[0020] 2) -C(O)R 15 ;

[0021] 3) -CH(OH)SO3R 16 ;

[0022] 4) -C(O)NR 13 R 14 and

[0023] 5) -C(O)C(O)NR 13 R 14 ;

[0024] R1, R2and R3are independently selected from:

[0025] 1) hydrogen;

[0026] 2) optionally substituted -C1-C8alkyl;

[0027] 3) optionally substituted -C2-C8alkenyl;

[0028] 4) optionally substituted -C2-C8alkynyl;

[0029] 5) optionally substituted -C3-C8cycloalkyl;

[0030] 6) optionally substituted 3- to 8-membered heterocycloalkyl;

[0031] 7) optionally substituted aryl;

[0032] 8) optionally substituted arylalkyl;

[0033] 9) optionally substituted heteroaryl; and

[0034] 10) optionally substituted heteroarylalkyl;

[0035] Alternatively, R1and R2together with the carbon atom to which they are attached form an optionally substituted 3- to 8-membered carbocyclo or an optionally substituted 3- to 8-membered heterocyclyl ring.

[0036] R4is hydrogen, optionally substituted -C1-C4alkyl, optionally substituted -C2-C4alkenyl or optionally substituted -C3-C6cycloalkyl.

[0037] R 11 and R 12 are each independently selected from:

[0038] 1) optionally substituted -C1-C8alkyl;

[0039] 2) optionally substituted -C2-C8alkenyl;

[0040] 3) optionally substituted -C2-C8alkynyl;

[0041] 4) optionally substituted -C3-C8cycloalkyl;

[0042] 5) optionally substituted 3- to 8-membered heterocycloalkyl;

[0043] 6) optionally substituted aryl;

[0044] 7) optionally substituted arylalkyl;

[0045] 8) optionally substituted heteroaryl; and

[0046] 9) optionally substituted heteroarylalkyl;

[0047] R 13 and R 14 are each, independently of one another, selected from:

[0048] 1) hydrogen;

[0049] 2) optionally substituted -C1-C8alkyl;

[0050] 3) optionally substituted -C2-C8alkenyl;

[0051] 4) optionally substituted -C2-C8alkynyl;

[0052] 5) optionally substituted -C3-C8cycloalkyl;

[0053] 6) optionally substituted 3- to 8-membered heterocycloalkyl;

[0054] 7) optionally substituted aryl;

[0055] 8) optionally substituted arylalkyl;

[0056] 9) optionally substituted heteroaryl; and

[0057] 10) optionally substituted heteroarylalkyl;

[0058] optionally, R 13 and R 14 together with the nitrogen atom to which they are attached form an optionally substituted 3- to 8-membered heterocyclyl ring;

[0059] R 15 is hydrogen, hydroxyl or optionally substituted -C1-C8alkyl; and

[0060] R 16is hydrogen or Na + .

[0061] In certain embodiments, the present application provides compounds of Formula (I) and pharmaceutically acceptable salts, esters and prodrugs thereof,

[0062]

[0063] wherein:

[0064] A is selected from:

[0065] 1) -R 11 ;

[0066] 2) -OR 12 ; and

[0067] 3) -NR 13 R 14 ;

[0068] B is optionally substituted aryl or optionally substituted heteroaryl;

[0069] X is selected from:

[0070] 1) -CN;

[0071] 2) -C(O)R 15 ;

[0072] 3) -CH(OH)SO3R 16 ;

[0073] 4) -C(O)NR 13 R 14 ; and

[0074] 5) -C(O)C(O)NR 13 R 14 ;

[0075] R1, R2and R3are independently selected from:

[0076] 1) hydrogen;

[0077] 2) optionally substituted -C1-C8alkyl;

[0078] 3) optionally substituted -C2-C8alkenyl;

[0079] 4) optionally substituted -C2-C8alkynyl;

[0080] 5) optionally substituted -C3-C8cycloalkyl;

[0081] 6) optionally substituted 3- to 8-membered heterocycloalkyl;

[0082] 7) optionally substituted aryl;

[0083] 8) optionally substituted arylalkyl;

[0084] 9) optionally substituted heteroaryl; and

[0085] 10) optionally substituted heteroarylalkyl;

[0086] Optionally, R1and R2together with the carbon atom to which they are attached form an optionally substituted 3- to 8-membered carbocyclyl ring or an optionally substituted 3- to 8-membered heterocyclyl ring.

[0087] R 11 and R 12 are each independently selected from:

[0088] 1) optionally substituted -C1-C8alkyl;

[0089] 2) optionally substituted -C2-C8alkenyl;

[0090] 3) optionally substituted -C2-C8alkynyl;

[0091] 4) optionally substituted -C3-C8cycloalkyl;

[0092] 5) optionally substituted 3- to 8-membered heterocycloalkyl;

[0093] 6) optionally substituted aryl;

[0094] 7) optionally substituted arylalkyl;

[0095] 8) optionally substituted heteroaryl; and

[0096] 9) optionally substituted heteroarylalkyl;

[0097] R 13 and R 14 are each independently selected from:

[0098] 1) hydrogen;

[0099] 2) optionally substituted -C1-C8alkyl;

[0100] 3) optionally substituted -C2-C8alkenyl;

[0101] 4) optionally substituted -C2-C8alkynyl;

[0102] 5) optionally substituted -C3-C8cycloalkyl;

[0103] 6) optionally substituted 3- to 8-membered heterocycloalkyl;

[0104] 7) optionally substituted aryl;

[0105] 8) optionally substituted arylalkyl;

[0106] 9) optionally substituted heteroaryl; and

[0107] 10) optionally substituted heteroarylalkyl;

[0108] R 13 and R 14 together with the nitrogen atom to which they are attached form an optionally substituted 3- to 8-membered heterocyclyl ring;

[0109] R 15 is hydrogen, hydroxyl, or optionally substituted -C1-C8 alkyl; and

[0110] R 16 is hydrogen or Na + . DETAILED DESCRIPTION

[0111] In one embodiment of the application, the application relates to a compound of formula (I) or formula (Ia) as described above, or a pharmaceutically acceptable salt thereof.

[0112] In one embodiment of the application, the compound of formula (Ia) can be represented as formula (Ia-A) or formula (Ia-B), or a pharmaceutically acceptable salt, ester or prodrug thereof:

[0113]

[0114] wherein A, B, X, R1, R2, R3, and R4 are as previously defined.

[0115] In a preferred embodiment, the stereochemical configuration of the compound of formula (Ia) is as represented by formula (Ia-A).

[0116] In one embodiment of the application, the compound of formula (I) can be represented as formula (I-A) or formula (I-B), or a pharmaceutically acceptable salt, ester or prodrug thereof:

[0117]

[0118] wherein A, B, X, R1, R2, and R3 are as previously defined.

[0119] In a preferred embodiment, the stereochemical configuration of the compound of formula (I) is as represented by formula (I-A).

[0120] In certain embodiments of a compound of Formula (I) or Formula (Ia), R1is hydrogen or optionally substituted -Ci-C4alkyl; optionally substituted -C3-C6cycloalkyl; optionally substituted aryl; optionally substituted arylalkyl; optionally substituted heteroarylalkyl. In certain embodiments of a compound of Formula (I) or Formula (Ia), R1is hydrogen or optionally substituted -Ci-C6alkyl; optionally substituted -C3-C6cycloalkyl; optionally substituted C3-C6cycloalkyl-Ci-C2-alkyl-; optionally substituted aryl; optionally substituted arylalkyl; optionally substituted heteroarylalkyl.

[0121] In certain embodiments of a compound of Formula (I) or Formula (Ia), R2is hydrogen or optionally substituted -Ci-C4alkyl; optionally substituted -C3-C6cycloalkyl; optionally substituted aryl; optionally substituted arylalkyl; optionally substituted heteroarylalkyl.

[0122] In certain embodiments of a compound of Formula (I) or Formula (Ia), R3is hydrogen or optionally substituted -Ci-C4alkyl; R4is hydrogen or or optionally substituted -Ci-C4alkyl.

[0123] In certain embodiments of a compound of Formula (I) or Formula (Ia), R3is hydrogen, -Me, -Et, -Pr, -i-Pr, -allyl, -CF3, -CD3, or cyclopropyl.

[0124] In certain embodiments of a compound of Formula (Ia), R4is hydrogen, -Me, -Et, -Pr, -i-Pr, -allyl, -CF3, or cyclopropyl.

[0125] In certain embodiments of a compound of Formula (I) or Formula (Ia), X is -CN.

[0126] In certain embodiments of a compound of Formula (I) or Formula (Ia), X is -C(O)H.

[0127] In certain embodiments of a compound of Formula (I) or Formula (Ia), X is -C(O)CH2OH, -C(O)CH2Cl, or -C(O)CH2F.

[0128] In certain embodiments of a compound of Formula (I) or Formula (Ia), X is -C(O)C(O)NR 13 R 14 , R 13 and R 14 are as previously defined.

[0129] In certain embodiments of the compounds of formula (I) or formula (la), A is derived from any of the following groups by removal of one hydrogen atom, and can be optionally substituted:

[0130]

[0131] In certain embodiments of the compounds of formula (I) or formula (la), A is selected from the group consisting of:

[0132]

[0133] Preferably, the substituents are independently selected from the group consisting of halogen, CN, NH2, optionally substituted -C1-C3alkoxy, optionally substituted -C1-C3alkyl, optionally substituted -C3-C6cycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl. Preferably, the number of substituents is 0 to 3.

[0134] In certain embodiments of the compounds of formula (I) or formula (la), A is selected from the group consisting of:

[0135]

[0136] In certain embodiments of the compounds of formula (I) or formula (la), A is selected from the group consisting of:

[0137]

[0138] Preferably, the substituents are independently selected from the group consisting of halogen, CN, NH2, optionally substituted -C1-C3alkoxy, optionally substituted -C1-C3alkyl, optionally substituted -C3-C6cycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl. Preferably, the number of substituents is 0 to 3.

[0139] In certain embodiments of the compounds of formula (I) or formula (la), A is selected from the group consisting of:

[0140]

[0141] Preferably, the substituents are independently selected from the group consisting of halogen, CN, NH2, optionally substituted -C1-C3alkoxy, optionally substituted -C1-C3alkyl, optionally substituted -C3-C6cycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl. Preferably, the number of substituents is 0 to 3.

[0142] In certain embodiments of the compounds of formula (I) or formula (la), B is selected from the group consisting of:

[0143]

[0144] In certain embodiments, the compound of Formula (Ia) is represented as Formula (Ia-1):

[0145]

[0146] wherein A, B, R1, R2, R4and X are as previously defined.

[0147] In certain embodiments, the compound of Formula (Ia) is represented as Formula (Ia-2):

[0148]

[0149] wherein A, B, R1, R3, R4and X are as previously defined.

[0150] In certain embodiments, the compound of Formula (Ia) is represented as Formula (Ia-3):

[0151]

[0152] wherein A, B, R1, R4and X are as previously defined.

[0153] In certain embodiments, the compound of Formula (I) is represented as Formula (I-1):

[0154]

[0155] wherein A, B, R1, R2and X are as previously defined.

[0156] In certain embodiments, the compound of Formula (I) is represented as Formula (I-2):

[0157]

[0158] wherein A, B, R1, R3and X are as previously defined.

[0159] In certain embodiments, the compound of Formula (I) is represented as Formula (I-3):

[0160]

[0161] wherein A, B, R1and X are as previously defined.

[0162] In certain embodiments, the compound of Formula (Ia) is represented as Formula (IIa):

[0163]

[0164] wherein A, R1, R2, R3, R4and X are as defined previously, and each R9is independently selected from:

[0165] 1) halo;

[0166] 2) -CN;

[0167] 3) -OR 13 ;

[0168] 4) -SR 13 ;

[0169] 5) -NR 13 R 14 ;

[0170] 6) -OC(O)NR 13 R 14 ;

[0171] 7) optionally substituted -C1-C6alkyl;

[0172] 8) optionally substituted -C3-C8cycloalkyl;

[0173] 9) optionally substituted 3- to 8-membered heterocycloalkyl;

[0174] 10) optionally substituted aryl; and

[0175] 11) optionally substituted heteroaryl;

[0176] and n is 0, 1, 2, 3 or 4.

[0177] In certain embodiments, the compound of Formula (I) is represented by Formula (II):

[0178]

[0179] wherein A, R1, R2, R3and X are as defined previously, and

[0180] each R9is independently selected from:

[0181] 1) halo;

[0182] 2) -CN;

[0183] 3) -OR 13 ;

[0184] 4) -SR 13 ;

[0185] 5) -NR 13 R 14 ;

[0186] 6) -OC(O)NR 13R 14 ;

[0187] 7) optionally substituted -C1-C6 alkyl;

[0188] 8) optionally substituted -C3-C8 cycloalkyl;

[0189] 9) optionally substituted 3- to 8-membered heterocycloalkyl;

[0190] 10) optionally substituted aryl; and

[0191] 11) optionally substituted heteroaryl;

[0192] and n is 0, 1, 2, 3 or 4.

[0193] In certain embodiments, each R9is independently selected from the group consisting of chloro, fluoro, methoxy and trifluoromethoxy.

[0194] In certain embodiments, the compound of formula (Ia) is represented by formula (IIIa-1):

[0195]

[0196] wherein A, R1, R3, R4, R9, n and X are as previously defined.

[0197] In certain embodiments, the compound of formula (Ia) is represented by formula (IIIa-2):

[0198]

[0199] wherein A, R1, R2, R4, R9, n and X are as previously defined.

[0200] In certain embodiments, the compound of formula (I) is represented by formula (III):

[0201]

[0202] wherein A, R1, R2, R9, n and X are as previously defined.

[0203] In certain embodiments, the compound of formula (Ia) is represented by any one of formulae (IVa-1) to (IVa-6):

[0204]

[0205] wherein A, B, R1, R2, R3, R4, R 13 and R 14 are as previously defined.

[0206] In certain embodiments, the compound of Formula (I) is represented by any one of Formulae (IV-1) to (IV-6):

[0207]

[0208] wherein A, B, R1, R2, R3, R 13 and R 14 are as previously defined.

[0209] In certain embodiments, the compound of Formula (la) is represented by any one of Formulae (Va-1) to (Va-6):

[0210]

[0211] wherein A, R1, R2, R3, R4, R9, R 13 , R 14 and n are as previously defined.

[0212] In certain embodiments, the compound of Formula (I) is represented by any one of Formulae (V-1) to (V-6):

[0213]

[0214] wherein A, R1, R2, R3, R9, R 13 , R 14 and n are as previously defined.

[0215] In certain embodiments, the compound of Formula (la) is represented by any one of Formulae (Via-1) to (Via-6):

[0216]

[0217] wherein A, R1, R3, R4, R9, R 13 , R 14 and n are as previously defined.

[0218] In certain embodiments, the compound of Formula (I) is represented by any one of Formulae (VI-1) to (VI-6):

[0219]

[0220] wherein A, R1, R3, R9, R 13 , R 14 and n are as previously defined.

[0221] In certain embodiments, the compound of Formula (I) is represented by any one of Formulae (VII-1) to (VII-5):

[0222]

[0223] wherein A, R1, R3, and X are as previously defined. Preferably, A is selected from the group consisting of:

[0224] R1is selected from the group consisting of:

[0225]

[0226] and X is selected from the group consisting of:

[0227]

[0228] In certain embodiments, the compound of Formula (I) is represented by any one of Formulas (VII-6) to (VII-9):

[0229]

[0230] wherein A, R1, R3, R9, and X are as previously defined. Preferably, A is selected from the group consisting of:

[0231]

[0232] R1is selected from the group consisting of:

[0233]

[0234] and X is selected from the group consisting of:

[0235]

[0236] In certain embodiments, the compound of Formula (I) is represented by any one of Formulas (VII-1a) to (VII-5a):

[0237]

[0238] wherein A, R1, R3, and X are as previously defined. Preferably, A is selected from the group consisting of:

[0239]

[0240] R1is selected from the group consisting of:

[0241]

[0242] and X is selected from the group consisting of:

[0243]

[0244] In certain embodiments, the compound of formula (I) is represented by any one of formulae (VII-6a) to (VII-9a):

[0245]

[0246] wherein A, R1, R3, R9 and X are as defined above. Preferably, A is selected from the group consisting of:

[0247]

[0248] R1is selected from the group consisting of:

[0249]

[0250] and X is selected from the group consisting of:

[0251]

[0252] In certain embodiments, the compound of formula (I) is represented by any one of formulae (VII-1) to (VII-9) and (VII-1a) to (VII-9a), wherein A is selected from the group consisting of:

[0253]

[0254] X is selected from the group consisting of:

[0255]

[0256] and R1is selected from the group consisting of:

[0257]

[0258] In certain embodiments, the compound of formula (Ia) is represented by any one of formulae (VIII-1) to (VIII-5):

[0259]

[0260] wherein A, X, R1, R3, R4, and R9 are as defined above.

[0261] In certain embodiments, the compound of formula (Ia) is represented by any one of formulae (VIII-1a) to (VIII-5a):

[0262]

[0263] wherein A, R1, R3, and R9 are as defined above.

[0264] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (IX-1) to (IX-5):

[0265]

[0266] wherein A, X, R1, R3, R4, and R9 are as previously defined.

[0267] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (IX-1a) to (IX-5a)

[0268]

[0269]

[0270] wherein A, R1, R3, and R9 are as previously defined.

[0271] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (VIII-1) to (VIII-5) and Formulas (IX-1) to (IX-5), wherein R3 is hydrogen, -Me, -Et, -Pr, -i-Pr, -allyl, -CF3, -CD3, or cyclopropyl; R4 is hydrogen, -Me, -Et, -Pr, -i-Pr, -allyl, -CF3, or cyclopropyl; R9 is halogen, -OCH3, -NH2, -CH3, or -CF3; and A is selected from the group consisting of:

[0272]

[0273] X is selected from the group consisting of:

[0274]

[0275] and R1 is selected from the group consisting of:

[0276]

[0277] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (X-1) to (X-3):

[0278]

[0279] wherein m is 0, 1, 2, 3, 4, or 5; v is 0, 1, or 2; R 10 is optionally substituted -C1-C4 alkyl or

[0280] optionally substituted -C3-C6 cycloalkyl; X, R1, R3, R4, R9, and n are as previously defined.

[0281] ​In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1) to (XI-3):

[0282]

[0283] wherein R1, R3, R4, R9, R 10 , m, n, and v are as defined previously.

[0284] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1a) to (XI-3a):

[0285]

[0286] wherein R1, R3, R9, R 10 , m, n, and v are as defined previously.

[0287] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1b) to (XI-3b):

[0288]

[0289] wherein R1, R3, R9, R 10 , m, n, and v are as defined previously.

[0290] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1) to (XI-3):

[0291]

[0292] wherein R1, R4, R9, R 10 , m, n, and v are as defined previously.

[0293] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1a) to (XI-6a):

[0294]

[0295] wherein R1, R9, R 10 , m, n, and v are as defined previously.

[0296] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XI-1b) to (XI-6b):

[0297]

[0298] wherein R1, R9, R 10 m, n, and v are as previously defined.

[0299] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XII-1c) to (XII-6c):

[0300]

[0301] wherein R1, R9, R 10 m, n, and v are as previously defined.

[0302] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XII-1c) to (XII-6d):

[0303]

[0304] wherein R1, R9, R 10 m, n, and v are as previously defined.

[0305] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XIII-1) to (XIII-6):

[0306]

[0307] wherein R4, R9, R 10 m, n, and v are as previously defined.

[0308] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XIII-1a) to (XIII-6a):

[0309]

[0310] wherein R9, R 10 m, n, and v are as previously defined.

[0311] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XIV-1) to (XIV-6):

[0312]

[0313] wherein R4, R9, R 10 m, n, and v are as previously defined.

[0314] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XIV-1a) to (XIV-6a):

[0315]

[0316] wherein R9, R 10 , m, n, and v are as previously defined.

[0317] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XVII-1) to (XVII-3):

[0318]

[0319] wherein A, R1, R4, R9, n, and X are as previously defined.

[0320] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XVI-1) to (XVI-3):

[0321]

[0322] wherein R1, R4, R9, R 10 , m, n, and v are as previously defined.

[0323] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XVII-1) to (XVII-3):

[0324]

[0325] wherein R9, R 10 , m, n, and v are as previously defined.

[0326] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulae (XVIII-1) to (XVIII-2):

[0327]

[0328] wherein one U is N or NR 13 , the other U is N, NR 13 , or CR 13 , the other U is N, NR 13 ,, or CR 13 , and the fourth U is O, S, N, NR 13 , or CR 13 ; each V is independently CR 13 or N; and R1, R3, R4, R9, n, and X are as previously defined.

[0329] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (XIX-1) to (XIX-9):

[0330]

[0331] wherein R1, R3, R4, R9, n, and X are as previously defined.

[0332] In certain embodiments, the compound of Formula (Ia) is represented by any one of Formulas (XX-1) to (XX-9):

[0333]

[0334] wherein R1, R3, and R9 are as previously defined.

[0335] Definitions

[0336] Listed below are definitions for various terms used to describe the present application. Unless otherwise defined, these definitions apply throughout this specification and claims.

[0337] As used herein, the term "aryl" refers to a monocyclic or bicyclic carbocyclic ring having at least one aromatic ring, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system including at least one aromatic ring. The polycyclic aryl can include fused rings, covalently linked rings, or a combination thereof.

[0338] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic aromatic group having one or more ring atoms selected from S, O, and N; and the remaining ring atoms are carbon, wherein any N or S contained in the ring can optionally be oxidized. Heteroaryl includes, but is not limited to, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazoline, thiazoline, oxazoline, isoxazoline, thiadiazoline, oxadiazoline, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazoline, benzoxazoline, and quinoxaline. A polycyclic heteroaryl can include fused rings, covalently linked rings, or a combination thereof.

[0339] According to the present application, the aromatic group can be substituted or unsubstituted.

[0340] As used herein, the term "bicyclic aryl" or "bicyclic heteroaryl" refers to a ring system consisting of two rings, wherein at least one ring is aromatic; and the two rings can be fused or covalently linked.

[0341] The term "alkyl" as used herein refers to a saturated, straight-chain or branched-chain hydrocarbon group. "C 1- The term "C4alkyl", "C 1- The term "C6alkyl", "C 1- The term "C8alkyl", "C 1- The term "C 12 The term "C2-C4alkyl", "C3-C4alkyl" or "C3-C6alkyl" refers to an alkyl group consisting of two to four, three to four or three to six carbon atoms, respectively. Examples of C1-C8alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, neopentyl, n-hexyl, heptyl, octyl groups.

[0342] The term "alkenyl" as used herein refers to a straight-chain or branched-chain hydrocarbon group having at least one carbon-carbon double bond derived by removal of one hydrogen atom. "C2-C8alkenyl", "C2-C 12 The term "C2-C4alkenyl", "C3-C4alkenyl" or "C3-C6alkenyl" refers to an alkenyl group consisting of two to four, three to four or three to six carbon atoms, respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 2-methyl-2-buten-2-yl, heptenyl, octenyl and the like.

[0343] The term "alkynyl" as used herein refers to a straight-chain or branched-chain hydrocarbon group having at least one carbon-carbon triple bond derived by removal of one hydrogen atom. "C2-C8alkynyl", "C2-C 12 The term "C2-C4alkynyl", "C3-C4alkynyl" or "C3-C6alkynyl" refers to an alkynyl group consisting of two to four, three to four or three to six carbon atoms, respectively. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 2-propynyl, 2-butynyl, heptynyl, octynyl and the like.

[0344] The term "cycloalkyl" as used herein refers to a monocyclic or polycyclic saturated carbocyclic compound, or a bicyclic or tricyclic fused, bridged, or spiro system, and the carbon atoms can optionally be substituted with oxygen, or optionally with exocyclic olefinic double bonds. Preferred cycloalkyl groups include C3-C 12 The term "C3-C6cycloalkyl", "C3-C8cycloalkyl" and "C4-C7cycloalkyl" refer to cycloalkyl groups consisting of three to six, three to eight, or four to seven carbon atoms, respectively. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. 12Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylenecyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexene, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonyl, and the like.

[0345] The term "cycloalkenyl" as used herein refers to a group derived from a monocyclic or polycyclic carbocyclic compound, or a bicyclic or tricyclic fused, bridged, or spiro system, and having at least one carbon-carbon double bond, and wherein the carbon atoms can optionally be substituted with oxygen, or optionally substituted with exogenous cycloalkene double bonds. Preferred cycloalkenyl groups include C3-C 12 cycloalkenyl, C3-C8cycloalkenyl, or C5-C7cycloalkenyl groups. C3-C 12 Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-2-enyl, bicyclo[4.2.1]non-3-en-12-yl, and the like.

[0346] The term "arylalkyl" as used herein refers to a functional group in which an alkyl chain is attached to an aryl group, for example, -CH2CH2-phenyl. The term "substituted arylalkyl" refers to arylalkyl functional groups in which the aryl group is substituted. Similarly, the term "heteroarylalkyl" refers to a functional group in which an alkyl chain is attached to a heteroaryl group. The term "substituted heteroarylalkyl" refers to heteroarylalkyl functional groups in which the heteroaryl group is substituted. Preferably, as used herein, arylalkyl is aryl-Ci-C6alkyl, and heteroarylalkyl is heteroaryl-Ci-C6alkyl.

[0347] The term "alkoxy," as used herein, alone or in combination with other terms, refers to an alkyl group having the indicated number of carbon atoms, as specified, linked via an oxygen atom to the rest of the molecule, for example, methoxy, ethoxy, 2-propoxy, 2-propoxy (isopropoxy), and higher analogs and isomers. Preferably, the alkoxy group is (C2-C3)alkoxy.

[0348] It should be understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, and cycloalkenyl groups described herein can also be aliphatic or alicyclic.

[0349] An "aliphatic" group is a non-aromatic group that can contain any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen, or other atoms, and optionally contains one or more units of unsaturation, e.g., a double and / or triple bond. Examples of aliphatic groups are functional groups such as, for example, 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, etc., groups comprising one or more than one aliphatic carbon-hydrogen group (optionally substituted) and certain other groups in which one or more than one carbon atom of the aliphatic carbon-hydrogen group (optionally substituted) is replaced by a functional group. The carbon atoms of an aliphatic group can be optionally oxygen substituted. An aliphatic group can be straight chained, branched, or cyclic, or a combination thereof, and preferably contains from about 1 to about 24 carbon atoms, more typically about 1 to about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, for example, as used herein, aliphatic groups also include, for example, alkoxyalkyl groups, polyalkoxyalkyl groups, e.g., polyalkylene oxides, polyamines and polyimines, for example, such aliphatic groups can be further optionally substituted.

[0350] The terms "heterocycloalkyl" and "heterocycle" can be used interchangeably and refer to a non-aromatic ring or a bicyclic or tricyclic group which is fused, spiro, or annexed, wherein (i) each ring contains at least one heteroatom, each independently selected from oxygen, sulfur, and nitrogen; (ii) each ring system can be saturated or unsaturated, (iii) the nitrogen and sulfur heteroatoms can be optionally oxidized, (iv) the nitrogen heteroatoms can be optionally quaternized, (v) any of the above rings can be fused to a benzene ring, and (vi) the remaining ring atoms are carbon atoms, which can be optionally substituted with oxygen or exogenous ring olefinic double bonds. Representative heterocycloalkyl groups include, but are not limited to, [l,3]dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolyl, isothiazolyl, quinoxalinyl, pyridazinyl, 2-azabicyclo[2.2.1]-heptene, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 2-oxa-7-azaspiro[4.4]nonyl, 7-oxanorborn-4-yl, and tetrahydrofuran. The heterocycloalkyl groups can be further substituted. The heteroaryl or heterocyclyl groups can be C-attached or N-attached (where possible).

[0351] It is understood that all alkyl, alkenyl, alkynyl, alicyclyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, aliphatic moieties, etc. described herein, when used as linkers to connect two or more groups or substituents (which can be the same or different), can be divalent or multivalent. The valence of any such group is readily determined by one of ordinary skill in the art from the context in which it appears.

[0352] The term "substituted" as used herein means that 1, 2, or 3 or more hydrogen atoms have been independently replaced by the following substituents, including but not limited to: -F, -Cl, -Br, -I, -OH, C 1- C 12 -alkyl; C2-C 12 -alkenyl, C2-C 12 -alkynyl, -C3-C 12 -cycloalkyl, protected hydroxyl, -NO2, -N3, -CN, -NH2, protected amino, oxygen, thio, -NH-C 1- C 12 -alkyl, -NH-C2-C8-alkenyl, -NH-C2-C8-alkynyl, -NH-C3-C 12- alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 1- C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C 1- C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C 1- C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C 1- C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocycloalkyl, -CO2-C 1- C 12 alkyl, -CO2-C2-C8alkenyl, -CO2-C2-C8alkynyl, CO2-C3-C 12 - cycloalkyl, -CO2-aryl, CO2-heteroaryl, CO2-heterocycloalkyl, -OCONH2, -OCONH-C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocycloalkyl, -NHC(O)H, -NHC(O)-C 1- C 12 - alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 - cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocycloalkyl, -NHCO2-C 1- 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-C 1- 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-C 1- 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-C 1- 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)-C 1- 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-C 1- 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)-C 1- C 12 - alkyl, -S(O)-C2-C8-alkenyl, -S(O)-C2-C8-alkynyl, -S(O)-C3-C 12- alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 1- C 12 - alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 12 - cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2- 1- C 12 - alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 12 - cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, 12 - cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, 1- C 12 - alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 12 - cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl, or methylsulfanyl-methyl. In certain embodiments, the substituents are independently of each other selected from the group consisting of halogen, preferably CI and F; C 1- C4-alkyl, preferably methyl and ethyl; halo-C 1- C4-alkyl, for example fluoromethyl, difluoromethyl, and trifluoromethyl; C2-C4-alkenyl; halo-C2-C4-alkenyl; C3-C6-cycloalkyl, for example cyclopropyl; C 1- C4-alkoxy, for example methoxy and ethoxy; halo-C 1- C4-alkoxy, for example fluoromethoxy, difluoromethoxy and trifluoromethoxy; acyl; -CN; -OH; NH2; C 1- C4-alkylamino; di(C 1- C4-alkyl)amino; and NO2. It will be appreciated that aryl, heteroaryl, alkyl, and the like can be further substituted. In some cases, each substituent of a substituted moiety is additionally optionally substituted with one or more groups, each group being independently selected from the group consisting of C 1-C4-alkyl; -CF3, -OCH3, -OCF3, -F, -Cl, -Br, -I, -OH, -NO2, -CN, and -NH2. Preferably, the substituted alkyl group can be further substituted with one or more halogen atoms, more preferably, one or more fluorine atoms or chlorine atoms.

[0353] The term "halo" or "halogen", alone or as part of another substituent, as used herein, means a fluorine, chlorine, bromine, or iodine atom. Preferred halogens are fluorine, chlorine, and bromine.

[0354] The term "optionally substituted", as used herein, means that the group in question can be substituted or unsubstituted. In one embodiment, the group in question is optionally substituted with zero substituents, e.g., the group in question is unsubstituted. In another embodiment, the group in question is substituted with one or more other groups, said other groups being individually and independently selected from the groups described herein.

[0355] The term "hydrogen" includes hydrogen and deuterium. In addition, the description of the molecules includes isotopes of the molecules, provided that the resulting compound is pharmaceutically acceptable.

[0356] The term "hydroxyl activating group", as used herein, refers to a labile chemical group known in the art that activates a hydroxyl group to be removed during a synthetic process, e.g., in a substitution or elimination reaction. Examples of hydroxyl activating groups include, but are not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate, and the like.

[0357] The term "activated hydroxyl group", as used herein, refers to a hydroxyl group activated by a hydroxyl activating group as defined above, e.g., the hydroxyl activating group includes mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate.

[0358] As used herein, the term "hydroxyl protecting group" refers to an unstable chemical group known in the art that protects hydroxyl groups from undesirable reactions during the synthetic process. Hydroxyl protecting groups as described herein can be selectively removed after the synthetic procedure. A summary of hydroxyl protecting groups known in the art is found in TH Greene and PG, SMWuts, Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl, isopropyloxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacyl, methoxyacyl, phenoxyacyl, benzoyl, methyl, tert-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, etc.

[0359] As used herein, the term "protected hydroxyl group" refers to a hydroxyl group protected by a hydroxyl protecting group as defined above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, and methoxymethyl.

[0360] As used herein, the term "hydroxyl precursor drug group" refers to a promoting group known in the art that can transiently alter the biological properties of a parent drug by covering or masking a hydroxyl group, thereby changing its physicochemical properties. Following the synthetic steps described herein, the hydroxyl precursor drug group must be able to revert to a hydroxyl group in vivo. Hydroxyl precursor drug groups known in the art are primarily described in Kenneth B. Sloan, Prodrugs, Topical and Ocular Drug Delivery , (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992).

[0361] As used herein, the term "amino protecting group" refers to an unstable chemical moiety known in the art that protects an amino group from undesirable reactions during the synthetic process. Following the synthetic procedure, the amino protecting group as described herein may be selectively removed. Amino protecting groups known in the art are primarily described in THGreene and PGMUTS. Protective Groups in Organic SynthesisExamples of amino protecting groups include, but are not limited to, methoxycarbonyl, tert-butoxycarbonyl, 12-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

[0362] The term "protected amino group" as used herein refers to an amino group protected by an amino protecting group as defined above.

[0363] The term "leaving group" refers to a functional group or atom which can be displaced by another functional group or atom in a substitution reaction, such as a nucleophilic substitution reaction. For example, representative leaving groups include chlorine, bromine, and iodine groups; sulfonate groups, such as methanesulfonate, toluenesulfonate, brosylate, nosylate, and the like; and acyloxy groups, such as acetoxy, trifluoroacetoxy, and the like.

[0364] The term "aprotic solvent" as used herein refers to a solvent which is relatively unreactive toward protons, i.e., has no proton donor activity. Examples include, but are not limited to, hydrocarbons such as hexane and toluene, halogenated hydrocarbons such as, for example, methylene chloride, ethylene chloride, chloroform, and the like; heterocyclic compounds such as, for example, tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethylether. Such solvents are well known to those skilled in the art and individual solvents or mixtures of solvents can be preferred for a particular compound and reaction conditions depending on factors such as solubility of reagents, reactivity of reagents, and preferred temperature ranges. Further discussion of aprotic solvents can be found in organic chemistry textbooks or monographs such as: Organic Solvents Physical Properties and Methods of Purification, 4thEdition, Volume II, Techniques of Chemistry Series, edited by John A. Riddick et al., John Wiley & Sons, New York, 1986

[0365] As used herein, the term "proton donating organic solvent" or "proton donating solvent" refers to solvents that tend to donate protons, such as an alcohol, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and individual solvents or mixtures of solvents can be preferred for a particular compound and reaction conditions, depending on factors such as solubility of reagents, reactivity of reagents, and preferred temperature ranges. Further discussion of proton donating solvents can be found in organic chemistry text books or monographs, such as: Organic Solvents Physical Properties and Methods of Purification, edited by John A. Riddick et al., Fourth Edition, Volume II, Techniques of Chemistry Series, John Wiley & Sons, New York, 1986.

[0366] The present application encompasses only those combinations of substituents and variables which are capable of giving rise to stable compounds. As used herein, the term "stable" refers to compounds which possess sufficient stability for manufacturing and handling without degrading to an unacceptably low level.

[0367] The synthesized compounds can be isolated from the reaction mixture and further purified by isolation methods, such as column chromatography, high pressure liquid chromatography, or recrystallization, and the like. As will be understood by those skilled in the art, further methods of synthesizing the compounds represented by the various chemical formulae herein will be apparent to those of ordinary skill in the art. In addition, the various synthetic steps can be performed in an alternate sequence or order without deviating from the spirit of the present application. As described herein, the methods of chemical transformations and protecting group methodologies (protection and deprotection) that can be effectively employed to synthesize the compounds described herein are known in the art, such as those described in R. Larock, Comprehensive Organic Transformations Protective Groups in Organic Synthesis, Second Edition, John Wiley & Sons (1991); L. Fieser and M. Fieser, Fires' Reagents for Organic Synthesis, Thirteenth Edition, John Wiley & Sons (1994); and L. Paquette, ed., Encyclopedia of Protective Groups in Organic Synthesis Protective Groups in Organic Synthesis, Second Edition, John Wiley & Sons (1991); L. Fieser and M. Fieser, Fires' Reagents for Organic Synthesis, Thirteenth Edition, John Wiley & Sons (1994); and L. Paquette, ed., Encyclopedia of Fieser and Fieser's Reagents for Organic SynthesisFisher and the Fischer Reagent for Organic Synthesis), Johns Wiley & Sons (1994); and Encyclopedia of Reagents for Organic Synthesis methods in the treatise, The Encyclopedia of Reagents for Organic Synthesis, 3rd ed.; John Wiley & Sons: New York, 1995, and related publications.

[0368] The term "subject" as used herein refers to an animal, preferably the animal is a mammal. More preferably, the mammal is a human. Subject can also refer to, for example, a dog, cat, horse, cow, pig, guinea pig, fish, bird, and the like.

[0369] The compounds of the present application can be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and can include, for example, increasing bioavailability by increasing oral uptake, increasing solubility to allow administration by injection, altering metabolism to allow for longer half-life, and altering excretion rates to allow for increased efficacy. The compounds of the present application can also be formulated as a pharmaceutical composition in combination with one or more other drugs as described herein.

[0370] The compounds described herein contain one or more asymmetric centers and thus exist in enantiomeric, diastereomeric, and other stereoisomeric forms. The absolute stereochemistry is specified where possible, using the (R)- or (S)-, or using the (D)- or (L)- notation, as appropriate. The present application is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared using, for example, the procedures described above, or by the resolution of racemic mixtures. The resolution procedures can be performed in the presence of a resolving agent, by chromatographic techniques or by repeated crystallization. Further details regarding resolution techniques are found in Jacques et al. Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds, unless specified otherwise, the compounds contain both the E and Z geometric isomers, and the present application includes each isomer either alone or in combination with the other. Similarly, all tautomeric forms of the compounds described herein are also intended to be included within the scope of the present application. The tautomeric forms can be cyclic or acyclic. The choice of a particular tautomeric form is only for convenience, and is not intended to designate a particular tautomeric form, unless otherwise indicated. Thus, a carbon-carbon double bond depicted in the present application as being in the trans form can alternatively be in the cis form, or a mixture of the two forms in any proportion.

[0371] Certain compounds of the present application can also exist in different stable conformational forms, which are separable. Isomerization between conformations can be allowed due to asymmetric single bonds, for example, torsional asymmetry restricted by steric hindrance or ring strain. The present application includes each conformational isomer of these compounds and mixtures thereof.

[0372] As used herein, the term "pharmaceutically acceptable salt" means a salt of a compound of the present application, which is within the scope of sound medical judgment of those skilled in the art, and which is suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 2-19 (1977). Salts can be prepared in situ during the final isolation and purification of the compounds of the application, or separately by reacting the appropriate compound of the application with an organic acid or base. Examples of pharmaceutically acceptable salts include, but are not limited to, nontoxic acid addition salts, e.g., salts of 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 known in the art such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl sulfonate, and aryl sulfonate.

[0373] In certain embodiments, the present application provides pharmaceutically acceptable esters of the compounds described herein. The term "pharmaceutically acceptable ester" as used herein refers to esters of the compounds that dissociate in vivo and include those that break down in human beings to yield the parent compound or a salt of the compound. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, in particular alkanoic, alkenanoic, cycloalkanoic and alkanedioic acids in which each aliphatic or olefinic moiety advantageously has not more than six carbon atoms. Examples of specific esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethyl succinates.

[0374] Pharmaceutical Compositions

[0375] The pharmaceutical compositions of the present application comprise a therapeutically effective amount of a compound of the present application, and one or more pharmaceutically acceptable carriers.

[0376] The term "pharmaceutically acceptable carrier" as used herein means a non-toxic, inert solid, semi-solid or liquid filler, diluent, capsule, or any type of formulation auxiliary of any type that is nontoxic to the subject. Examples of materials which can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols, such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other nontoxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0377] The pharmaceutical compositions of the present application can be administered in a form suitable to the delivery of the compounds to the site of desired action. Preferably, the pharmaceutical compositions of the present application are administered in oral or injectable form. The pharmaceutical compositions of the present application can comprise any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation can be adjusted with pharmaceutically-acceptable acids, bases or buffers to make the formulation more stable, either to the compound itself or to the form of the release of the compound. As used herein, the term parenteral includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques.

[0378] Liquid forms in which the compounds can be given orally include aqueous solutions, emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid formulations can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, among others, mixed solvents, and combinations thereof.

[0379] Injectable preparations, which can be formulated in accordance with known techniques, using suitable dispersing or wetting agents and suspending agents can be sterile injectable aqueous or oleaginous suspensions. The sterile injectable preparation can also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol alone or in combination with a nontoxic parenterally acceptable diluent. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

[0380] The injectable formulations can be sterilized by, for example, filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0381] To prolong in vivo action, it is often desirable to slow absorption of the drug from injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of such a drug then depends upon its rate of dissolution which, in turn, can depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer, and the particular polymer used, the rate of drug release can be controlled. Other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

[0382] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this application with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0383] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or: a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form can also comprise buffering agents.

[0384] Solid compositions of a similar type can also be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols, and the like.

[0385] The active compounds can be prepared in the form of a microemulsion with one or more excipients mentioned above. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared using binding agents such as microcrystalline cellulose, pre-gelatinized starch, gelatin, acacia, starch, and the like, together with excipients such as lactose, magnesium stearate, stearic acid, sodium saccharin, talc, magnesium carbonate, and the like. Solid dosage forms can also comprise, as is normal practice, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; flavoring agents; preservatives; colorants; and similar agents to improve flavor, appearance, and shelf life. If desired, disintegrating or solubilizing agents can be present, such as starches, menthol, agar-agar, beta-lactose, calcium phosphate, polyvinylpyrrolidone, sodium starch glycolate, and the like. Tablets can be uncoated or can be coated by known techniques including microencapsulation.

[0386] Formulations of the compounds of the application for topical or transdermal administration can include ointments, pastes, creams, lotions, jellies, powders, solutions, sprays, inhalants, or patches. The active component is admixed with a pharmaceutical acceptable carrier, such as a preservative, or buffer, if desired. Ophthalmic formulations, eardrops, eye drops, powders, and solutions are contemplated as being within the scope of the application.

[0387] The ointments, pastes, creams, and jellies can contain, in addition to the active compound of this application, excipients such as animal and vegetable fats and oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0388] The powders and sprays can contain, in addition to the compound of this application, excipients such as lactose, talc, aluminim hydroxide, calcium silicates, and polyamide powder or mixtures of these substances. The sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

[0389] Transdermal patches have the added advantage of controlling the rates of release of the compound into the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0390] For pulmonary delivery, the therapeutic compositions of the present application are formulated to be taken in solid form by the patient or are formulated in a liquid particulate form for direct administration to the respiratory system, e.g., by inhalation. Solid or liquid particulate forms of the active compounds of the present application prepared in accordance with the present application include particles of respirable size: that is, small enough to be taken into the mouth, inhaled into the bronchial tube and into the alveoli of the lungs. Nebulized therapy, and in particular, nebulized antibiotics, are known in the art (see, e.g., U.S. Patent No. 5,767,068 to Van Devanter et al., U.S. Patent No. 5,508,269 to Smith et al., and WO 98 / 43650 to Montgomery et al., which are all incorporated herein by reference).

[0391] Antiviral Activity

[0392] In certain embodiments, the present application provides a method for treating or preventing a viral infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. The viral infection is preferably a coronavirus infection. In certain embodiments, the coronavirus is SARS-CoV-1, SARS-CoV-2, or MERS-CoV. Preferably, the coronavirus is SARS-CoV-2.

[0393] An inhibitory amount or dose of the compounds described herein can range from about 0.01 mg / kg to about 500 mg / kg, or alternatively from about 1 to about 50 mg / kg. The inhibitory amount or dose will also vary depending on the route of administration, and the possibility of co-usage with other agents.

[0394] In accordance with the methods of treatment described herein, a viral infection, disorder in a host, e.g., a human or another animal, is treated or prevented by administering to the patient a therapeutically effective amount of a compound described herein, in a dosage and for a time necessary to achieve the desired result.

[0395] The "therapeutic effective dose" of the compounds described in this invention refers to a dose of the compound that provides a therapeutic effect to the host receiving treatment with a reasonable benefit / risk ratio, wherein the benefit / risk ratio is applicable to any drug treatment. The therapeutic effect can be objective (i.e., measurable by some test or marker) or subjective (i.e., the host exhibits signs or sensations of an effect). The effective dose of the compounds described above can be found in the range of about 0.1 mg / kg to about 500 mg / kg, preferably in the range of about 1 to about 50 mg / kg. The effective dose will also vary depending on the route of administration and the possibility of co-administration with other formulations. However, it should be understood that the total daily dosage of the compounds and compositions described in this invention will be determined by the attending physician within a reasonable medical judgment. For any given patient, the level of a specific therapeutically effective dose will depend on a variety of factors, including the disorder requiring treatment and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0396] The total daily dose of the compounds described in this invention, administered to humans or other animals in a single dose or in fractionated doses, can be, for example, from 0.01 to 50 mg / kg body weight or more, typically from 0.1 to 25 mg / kg body weight. A single-dose composition may contain such a dose or submultiples that constitute this daily dose. Generally, treatment regimens according to this invention involve administering approximately 10 mg to approximately 1000 mg of the compounds described in this invention daily in a single or multiple doses to a patient requiring such treatment.

[0397] The compounds described herein can be administered, for example, in a form suitable for injection, intravenously, intraarterially, subdermally, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in an ophthalmic preparation, or by inhalation, in dosage unit formulations containing non-toxic excipients that are suitable for such administration. The compounds described herein can be administered at dosages, and / or with dosing regimens, consistent with such administration in amounts that will yield suitable systemic levels of the active compounds. As described herein, the methods contemplate administration of an effective amount of the compound or combination of compounds to achieve the desired or stated effect. Generally, the pharmaceutical compositions described herein will be administered from about 1 to about 6 times per day or alternatively, as a single dosage schedule upon periodic intervals. Such administration can be used as a chronic or acute therapy. The dosage regimen will be dependent on the dosage of the active ingredients employed, as well as the frequency and time course of administration. The active ingredients can be administered in a single dose, or multiple doses, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 doses. A typical formulation might contain from about 5% to about 95% active compound (w / w). Alternatively, such formulations can contain from about 20% to about 80% active compound.

[0398] Lower or higher doses than those recited above can be required. Specific dosage and treatment regimens will depend on a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition, or symptoms, the patient's disposition to the disease, condition, or symptoms, and the judgment of the treating physician.

[0399] When the patient's condition has been alleviated to the desired level, administration of a maintenance dose of the compound, composition or combination of the present application can be initiated. Subsequently, the dosage or frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved condition is maintained, up to the complete disappearance of the symptoms. However, should disease symptoms flare up at any time, the patient can resume the intermittent treatment on a long-term basis.

[0400] Combination Therapy and Switch Therapy

[0401] The compounds of the present application can be used in combination with one or more antiviral agents or anti-inflammatory agents that are useful in the prevention or treatment of viral diseases or related pathophysiology. Thus, the compounds of the present application, and salts, solvates, or other pharmaceutically acceptable derivatives thereof, can be used alone or in conjunction with other antiviral or anti-inflammatory therapies. The compounds and pharmaceutically acceptable salts thereof, herein can be used in combination with one or more other agents that are effective in the prevention or treatment of respiratory diseases, inflammatory diseases, autoimmune diseases, such as antihistamines, corticosteroids (e.g., fluticasone propionate, fluticasone furanoate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furanoate, triamcinolone acetonide, fluocinolone acetonide), non-steroidal anti-inflammatory drugs, leukotriene modulators (e.g., montelukast, zafirlukast, pranlukast), tryptase inhibitors, IKK2 inhibitors, p38 inhibitors, Syk inhibitors, protease inhibitors, such as elastase inhibitors, integrin antagonists (e.g., beta-2 integrin antagonists), adenosine A2a agonists, mediator release inhibitors (e.g., zyflo, DP1 antagonists, DP2 antagonists, PI3K delta inhibitors, ITK inhibitors, LP (iodophosphoric acid) inhibitors, or FLAP (5-lipoxygenase activating protein) inhibitors (e.g., 3-(3-(tert-butylthio)-l-(4-(6-ethoxypyridin-3-yl)benzyl)-5-((5- ethylpyridin-2-yl)methoxy)-lH-indol-2-yl)-2,2-dimethylpropanoic acid sodium salt), bronchodilators (e.g., muscarinic antagonists, beta-2 agonists), methotrexate, and the like; monoclonal antibody therapies such as anti-lgE, anti-TNF, anti-IL-5, anti-IL-6, anti-IL-12, anti-IL-1, and the like; cytokine receptor therapies such as etanercept and the like; antigen non-specific immunotherapies (e.g., interferons or other cytokines / chemokines, chemokine receptor modulators such as CCR3, CCR4, or CXCR2 antagonists, other cytokine / chemokine agonists or antagonists, TLR agonists, and the like), appropriate anti-infective agents, including antibiotic agents, antifungal agents, antihelminthic agents, antimalarial agents, antiprotozoal agents, antitubercular agents, and antiviral agents, including those described at https: / / www.drugs.com / drug-class / anti-infectives.html. In general, combination therapy is often preferred over alternative therapies, as combination therapy can simultaneously exert multiple pressures on the virus.

[0402] When the compositions of the present application comprise a combination of a compound according to the formulae described herein and one or more other therapeutic or prophylactic agents, both the compound and other agent are present in dosage form(s) that are typically administered in a single dosing regimen, and more preferably, at dosage levels that are between 1 and 100% of the dosage normally employed when the agents are used alone, and most preferably, between 5% and 95% of the dosage normally employed. The other agent(s) can be administered separately, as part of a multiple dosage regimen, from the compound of the present application. Alternatively, these agents can be part of a single dosage form, in which case they can be physically combined with the compound of the present application in a single composition.

[0403] "Other therapeutic or prophylactic agents" include, but are not limited to, immunotherapies (e.g., interferons), therapeutic vaccines, anti-fibrotic agents, anti-inflammatory agents, e.g., corticosteroids or non-steroidal anti-inflammatory drugs (NSAIDs), bronchodilators, e.g., beta-2 adrenergic agonists and xanthines (e.g., theophylline), mucolytic agents, antimuscarinics, anti-leukotrienes, cell adhesion inhibitors (e.g., ICAM antagonists), antioxidants (e.g., N-acylcysteines), cytokine agonists, cytokine antagonists, lung surfactants, and / or antibacterial and antiviral agents (e.g., ribavirin and amantadine). The compositions according to the present application can also be used in conjunction with gene replacement therapy.

[0404] Abbreviations

[0405] The following abbreviations will be used in the description of the schemes and examples hereinafter: Ac refers to acyl; AcOH refers to acetic acid; Boc2O refers to di-tert-butyl-dicarbonate; Boc refers to tert-butoxycarbonyl; Bz refers to benzoyl; Bn refers to benzyl; t-BuOK refers to potassium tert-butoxide; Brine refers to aqueous sodium chloride; CDI refers to carbonyldiimidazole; DCM or CH2Cl2refers to dichloromethane; CH3refers to methyl; CH3CN refers to acetonitrile; Cs2CO3refers to cesium carbonate; CuCl refers to copper (I) chloride; CuI refers to copper (I) iodide; dba refers to dibenzylideneacetone; DBU refers to 1,8-diazabicycloundec-7-ene; DEAD refers to diethylazodicarboxylate; DIAD refers to diisopropyl azodicarboxylate; DIPEA or (i-Pr)2EtN refers to N,N,-diisopropylethylamine; DMP or the Dess-Martin oxidant refers to 1,1,2-cis(acetyloxy)-1,2-dihydro-1,2-benziodoxol-3-(1H)-one; DMAP refers to 4-dimethylamino-pyridine; DME refers to 1,2-dimethoxyethane; DMF refers to N,N-dimethylformamide; DMSO refers to dimethylsulfoxide; EtOAc refers to ethyl acetate; EtOH refers to ethanol; Et2O refers to diethyl ether; HATU refers to O-(7-azabenzotriazol-2-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; HCl refers to hydrochloric acid; K2CO3refers to potassium carbonate; n-BuLi refers to n-butyllithium; DDQ refers to 2,3-dichloro-5,6-dicyano-1,4-benzoquinone; LDA refers to lithium diisopropylamide; LiTMP refers to lithium 2,2,6,6-tetramethyl-piperidide; MeOH refers to methanol; Mg refers to magnesium; MOM refers to methoxymethyl; Ms refers to methanesulfonyl or -SO2-CH3; NaHMDS refers to sodium bis(trimethylsilyl)amide; NaCl refers to sodium chloride; NaH refers to sodium hydride; NaHCO3refers to sodium bicarbonate or sodium hydrogen carbonate; Na2CO3refers to sodium carbonate; NaOH refers to sodium hydroxide; Na2SO4refers to sodium sulfate; NaHSO3refers to sodium sulfite or sodium bisulfite; Na2S2O3refers to sodium thiosulfate; NH2NH2refers to aqueous hydrazine; NH4Cl refers to ammonium chloride; Ni refers to nickel; OH refers to hydroxyl; OsO4refers to osmium tetroxide; OTf refers to trifluoromethylsulfonate; PPA refers to polyphosphoric acid; PTSA refers to p-toluenesulfonic acid; PPTS refers to pyridinium p-toluenesulfonate; TBAF refers to tetrabutylammonium fluoride; TEA or Et3N refers to triethylamine; TES refers to triethylsilyl; TESCl refers to triethylsilyl chloride; TESOTf refers to silyl trifluoromethanesulfonate; TFA refers to trifluoroacetic acid;THF means tetrahydrofuran; TMEDA means N,N,N',N'-tetramethylethylenediamine; TPP or PPh3 means triphenyl-phosphine; Tos or Ts means p-toluenesulfonyl or -SO2-C6H4CH3; Ts2O means p-toluenesulfonic anhydride or p-toluenesulfonyl anhydride; TsOH means p-toluenesulfonic acid; Pd means palladium; Ph means phenyl; Pd2(dba)3 means tris(dibenzylideneacetone)dipalladium(0); Pd(PPh3)4 means tetrakis(triphenylphosphine)- palladium(0); PdCl2(PPh3)2 means trans-dichloro-(triphenylphosphine)palladium(II); Pt means platinum; Rh means rhodium; rt means room temperature; Ru means ruthenium; TBS means tert-butyldimethylsilyl; TMS means trimethylsilyl; and TMSCl means trimethylsilyl chloride.

[0406] Synthetic Procedures

[0407] The compounds and methods of the present application will be better understood in connection with the following synthetic schemes that illustrate the methods of making the compounds of the present application, which are intended merely as an exemplification of the present application and are not intended to limit the scope of the application. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications including, without limitation, those related to the chemical structures, substituents, derivatives and / or methods of the application can be made without departing from the spirit and scope of the application and are intended to be covered by the following claims.

[0408] Scheme 1

[0409]

[0410] Scheme 1 illustrates a general method for preparing compounds of formula (IV-1) from amino ester compounds (X-1), wherein B is as previously defined and PG1is C1-C4alkyl or Bn. Treatment of amine (X-1) with formaldehyde gives the cyclized amine (X-2) which is converted to (X-3) using an appropriate protecting group PG2(e.g. Boc). Treatment of (X-3) with NBS in a solvent containing AcOH at low temperature provides the rearranged spiro pyrrolidine derivative (X-4). Examples of this transformation sequence have been reported in (Pellegrini C. et al. "Synthesis of the Oxindole Alkaloid (-)-Horsfiline" Tetrahedron Asymmetry, 1994, vol. 5, No. 10, pp 1979-1992; Efremov, I. V. et al. "Discovery and Optimization of a Novel spiral Pyrrolidine Inhibitor of β-Secretase (BACE1) through Fragment-Based Drug Design" Journal of Medicinal Chemistry, 2012, 55, 9069-9088). Treatment of ester (X-4) with NH3(e.g. ammonia in methanol, NH3OH, etc.) gives the amide compound (X-5) which is converted to the amine compound (X-6) by deprotection of the group PG2(e.g. TFA, HC1, etc.). Condensation of amine (X-6) with acid (X-7), wherein A, R1, R2and R3are as previously defined, is accomplished under amide coupling conditions (e.g. HATU, EDC, DCC, etc.) to yield the amide compound (X-8). Amide (X-8) is converted to the nitrile compound (IV-1) under dehydrating conditions, for example, but not limited to, TFAA / Et3N, or Pd(OCOCF3)2 / Cl2CHCN, boron reagent or T3P.

[0411] Alternatively, condensation of amine (X-6) with acid (X-9), wherein R1, R2and R3are as previously defined and PG3is an appropriate protecting group (e.g. Cbz), is accomplished under amide coupling conditions (e.g. HATU, EDC, DCC, etc.) to yield the amide compound (X-10). Removal of PG3(e.g. hydrogenation) provides the amine compound (X-11). Condensation of amine (X-11) with acid (A-COOH), wherein A is as previously defined, is accomplished under amide coupling conditions (e.g. HATU, EDC, DCC, etc.) or acyl lactone generating conditions (e.g. Ghosez reagent) to yield the amide compound (X-8).

[0412] Scheme 2

[0413]

[0414] Scheme 2 depicts a general method for the synthesis of aldehyde compounds of Formula (IV-2), wherein A, R1, R2, R3, and B are as previously defined. Reduction of ester compounds of Formula (X-4) with a reducing agent, such as, but not limited to, LiBH4, NaBH4, or DIBAL-H, produces alcohol compounds (XI-1), wherein B, PG1and PG2are as previously defined. Removal of the protecting group PG2(e.g., Boc) (XI-1) under acidic conditions, such as, but not limited to, TFA, HCl, formic acid, TMSOTf / dimethylpyridine, and the like. Coupling of amine compounds (XI-2) with acid compounds (X-7) using a coupling agent (e.g., HATU, EDC, or DCC) produces compounds (XI-3), wherein A, R1, R2, and R3are as previously defined. Oxidation of the alcohol compounds (XI-3) with mild oxidizing agents, such as DMSO / Ac2O, Dess-Martin Oxidizer, IBX, SO3-pyridine / DMSO / Et3N, produces aldehyde compounds (IV-2).

[0415] Scheme 3

[0416]

[0417] Scheme 3 shows a general method for the synthesis of hydroxymethyl ketone compounds of Formula (IV-3). Hydrolysis of ester compounds (X-4) produces acid compounds (XII-1), wherein B, PG1and PG2are as previously defined. Coupling of acid compounds (XII)-1 with N, O-dimethylhydroxylamine using reagents such as HATU, EDC, DCC, and the like can produce amide (XII-2). Treatment of amide (XII-2) with an organometallic reagent generated from BOM-Cl, Mg, and HgCl2at low temperature (about -60 °C) produces ketone compounds (XII-3). Removal of PG2(e.g., PTSA if PG2is BOC) provides amine compounds (XII-4). Coupling of amine (XII-4) with acid (X-7) using amide coupling reagents (e.g., HATU, EDC, DCC, and the like) produces compounds (XII-5), wherein A, R1, R2, and R3are as previously defined. Removal of the benzyl group in (XII-5) under hydrogenation conditions (Pd / C, H2) produces compounds of Formula (IV-3).

[0418] Scheme 4

[0419]

[0420] Scheme 4 illustrates a general method for the synthesis of chloromethyl ketone compounds of formula (IV-4). Treatment of ester compound (X-4) with an organometallic reagent generated from ICH2Cl and an appropriate base (e.g., LDA, MeLi / LiBr, or BuLi) yields chloroketone compound (XIII-1). Removal of PG2(e.g., PTSA if PG2is BOC) yields amine compound (XIII-2). Coupling of amine (XIII-2) with acid (X-7) using a coupling reagent (e.g., HATU, EDC, DCC, etc.) yields compound (IV-4), wherein A, R1, R2, and R3are as previously defined.

[0421] Scheme 5

[0422]

[0423] Scheme 5 illustrates a general method for the synthesis of chloromethyl ketone compounds of formula (IV-5). Pd catalyzed hydrogenation of the Bn group of compound (XII-3) yields alcohol compound (XIV-1). In some cases, e.g., SF4, Tf2O / dimethylpyridine / TBAF, C4F9SO2F / HF-Et3N, etc., alcohol (XIV-1) is converted to fluoromethyl ketone compound (XIV-2). Removal of PG2(e.g., PTSA if PG2is BOC) yields amine compound (XIV-3). Coupling of amine (XIV-3) with acid (X-7) using an amine coupling reagent (e.g., HATU, EDC, DCC, etc.) yields compound (IV-5), wherein A, R2, and R3are as previously defined.

[0424] Scheme 6

[0425]

[0426] Scheme 6 illustrates a general method for the synthesis of α-ketoamide compounds of formula (IV-6). Treatment of aldehyde compound of formula (IV-2), wherein A, R1, R2, R3, and B are as previously defined, with isonitrile compound (XV-1) (wherein R 13 is as previously defined) yields α-hydroxyamide (XV-2). Oxidation of compound (XV-2) with a suitable oxidizing agent, e.g., Dess-Martin oxidizing reagent, (COCl)2 / DMSO / Et3N, PCC, SO3-pyridine / DMSO / Et3N, etc., yields α-ketoamide compound (IV-6).

[0427] Scheme 7

[0428]

[0429] Alternatively, nitrile compounds (IV-1) can be synthesized from aldehyde compounds (IV-2) using the method shown in Scheme 7. Condensation of aldehyde (IV-2) with hydroxyl benzylamine in a suitable solvent (e.g., dimethyl sulfoxide, i-proline, pyridine, etc.) yields oxime compounds (XVI-1). Treatment of oxime compounds (XVI-1) under acid catalyzed dehydration conditions (e.g., (Cu(OAc)2 / MeCN, HCl, etc.) yields nitrile compounds (IV-1).

[0430] Scheme 8

[0431]

[0432] Scheme 8 shows a general method for synthesizing functionalized helical ring of formula XX-2 (Q1 is defined as halogen or optionally substituted alkyl). Treatment of a cyclic compound of helix formula XX-1 (wherein B, PG1 and PG2 are as previously defined) with an electrophilic reagent including but not limited to: sulfonyl chloride, N-chlorosuccinimide, N-bromosuccinimide, SelectFluor or NFSI, can yield functionalized helical ring XX-2.

[0433] Examples

[0434] The compounds and methods of the present application can be better understood in connection with the following examples, which are intended as illustrative only and not limiting of the scope of the application. Various modifications and changes can be made thereto by those of ordinary skill in the art which follow in the principles of the application and have the intended breadth of the scope of the application, including but not limited to variations and modifications involving chemical structures, substituents, derivatives, formulations and / or methods, without departing from the spirit and scope of the application as defined in the appended claims.

[0435] General Conditions:

[0436] Mass spectra were run on LC-MS systems using electrospray ionization. An Agilent 1290 Infinity II system with Agilent 6120 Quadrupole Detector was used. Spectra were obtained using a ZORBAX Eclipse XDB-C18 column (4.6 x 30 mm, 1.8 micron). Spectra were obtained at 298 K using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) as mobile phases. Spectra were obtained with the following solvent gradient: 0-1.5 min 5% (B), 1.5-4.5 min 5-95% (B) and 95% (B) from 4.5-6 min. The solvent flow rate was 1.2 mL / min. Compounds were detected at 210 nm and 254 nm wavelengths. [M+H]+ refers to the monoisotopic molecular weight.

[0437] Nuclear magnetic resonance spectra were run on a Bruker 400 MHz spectrometer. Spectra were measured at 298 K and the solvent peak was used as a reference. Chemical shifts for1H NMR are reported in parts per million (ppm).

[0438] Compounds were purified by reverse phase high performance liquid chromatography (RPHPLC) using a Gilson GX-281 automated liquid handling system. Compounds were purified on a Phenomenex Kinetex EVO C18 column (250 x 21.2 mm, 5 microns). Compounds were purified using a gradient elution between 0% and 100% of water (A) and acetonitrile (B) as mobile phase at 298 K, unless otherwise specified. The solvent flow rate was 20 ml / min and compounds were detected at a wavelength of 254 nm.

[0439] Alternatively, compounds were purified by normal phase liquid chromatography (NPLC) using a Teledyne ISCO Combiflash purification system. Compounds were purified on a REDISEP silica gel column. Compounds were purified at 298 K and detected at a wavelength of 254 nm.

[0440] Example 1

[0441]

[0442] Step 1-1

[0443] (S)-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole-3-carboxylic acid methyl ester hydrochloride (500 mg, 1.875 mmol) was dissolved in CH2Cl2(10 ml). Triethylamine (523 μΐ, 3.75 mmol) and a 2.0 M solution of di-tert-butyl dicarbonate in DCM (1031 μΐ, 2.062 mmol) were added. The reaction mixture was stirred at room temperature for 3 h, quenched with saturated NaHC03, and extracted with DCM. The organic layer was washed with brine, dried over MgS04, and concentrated in vacuo. The residue was purified on a silica gel column with 0-30% ethyl acetate / cyclohexane to yield compound (1-1) (578 mg, 1.749 mmol, 93% yield).

[0444] Step 1-2

[0445] Compound (1-1) was dissolved in THF (15 ml), AcOH (10 ml), and water (10 ml). The solution was cooled to -15 °C. A solution of NBS (328 mg, 1.843 mmol) in THF (5 mL) was added dropwise. The reaction mixture was slowly warmed to 5 °C over 1 h. The reaction was quenched with Na2S03and saturated NaHC03and extracted with DCM (2x). The organic layer was washed with brine, dried over MgS04and concentrated in vacuo. The residue was purified on a silica gel column with 0-50% ethyl acetate / cyclohexane to yield compound (1-2) (328 mg, 0.947 mmol, 53.9% yield).

[0446] Step 1-3

[0447] Compound (1-2) (328 mg, 0.947 mmol) was dissolved in MeOH (3 ml). A solution of 7 N ammonium in MeOH (5 mL, 35.0 mmol) was added. The reaction mixture was stirred at room temperature for 5 days. The solvent was removed in vacuo. The residue was purified on a silica gel column with 0-10% MeOH / DCM and on a C18 column with 0-50% MeCN / H20 to yield compound (1-3) (101 mg, 0.305 mmol, 32.2% yield).

[0448] Step 1-4

[0449] Compound (1-3) (100 mg, 0.302 mmol) was dissolved in DCM and trifluoroacetic acid (232 μΐ, 3.02 mmol) was added. The reaction mixture was stirred at 0 °C for 1 h and then at room temperature for 2 h. DCM (10 mL) and toluene (10 mL) were added. The solvent was removed in vacuo. The residue was dissolved in MeOH and 1 M HC1 (0.6 mL, 2 eq) was added. The solvent was removed. The resulting compound (1-4) (91 mg, 0.340 mmol, quantitative yield) was used in the next step.

[0450] Step 1-5

[0451] Compound (1-4) (15 mg, 0.056 mmol) and ((benzyloxy)carbonyl)-L- leucine (14.87 mg, 0.056 mmol) were dissolved in THF (0.5 ml) and DMF (0.1 ml). DIPEA (30.0 μΐ, 0.168 mmol) and HATU (21.30 mg, 0.056 mmol) were added. The reaction mixture was stirred at room temperature for 20 min, quenched with water, and extracted with EtOAc (2x). The organic layer was loaded on a silica gel column and eluted with 0-70% acetone / cyclohexane to yield compound (1-5) (15 mg, 0.031 mmol, 55.9% yield).

[0452] Step 1-6

[0453] Compound (1-5) (60 mg, 0.125 mmol) was dissolved in DCM (1.254 ml) (not dissolved). Triethylamine (140 μΐ, 1.003 mmol) and TFAA (70.8 μΐ, 0.502 mmol) were added. The reaction mixture was stirred at room temperature for 30 min. The reaction was diluted with DCM and quenched with saturated NaHC03. The organic layer was loaded on a silica gel column and eluted with 0-50% acetone / cyclohexane, then on a preparative-HPLC with 20-85% MeCN / H20 with 0.1% formic acid to yield Example 1 (14 mg, 0.056 mmol) as a white powder. 1 H NMR (400 MHz, Acetone-de) δ 9.70 (s, 1H), 7.42 - 7.31 (m, 5H), 7.28 (td, J = 7.7, 1.3 Hz, 1H), 7.12 (d, J = 7.4 Hz, 1H), 7.04 - 6.96 (m, 2H), 6.65 (d, J = 8.3 Hz, 1H), 5.17 (t, J = 8.3 Hz, 1H), 5.06 - 4.94 (m, 2H), 4.48 (td, J = 9.0, 5.0 Hz, 1H), 4.26 (d, J = 10.4 Hz, 1H), 3.99 (d, J = 10.3 Hz, 1H), 2.78 - 2.63 (m, 2H), 1.80 (dd, J = 13.8, 6.9 Hz, 1H), 1.74 - 1.56 (m, 2H), 0.96 (dd, J = 8.7, 6.6 Hz, 6H). [M+Na] m / e 483.18.

[0454] The following examples were prepared using similar protocols as described above.

[0455]

[0456]

[0457]

[0458]

[0459] Examples 11 & 12

[0460]

[0461]

[0462] Step 1

[0463] 4-Methoxy-lH-indole-2-carboxylic acid (1 g, 5.23 mmol) was dissolved in THF (25 mL) and ethyl L-leucine hydrochloride (1.024 g, 5.23 mmol), hunig's base (2.3 mL, 13.08 mmol), DMAP (0.032 g, 0.262 mmol), and HATU (2.0 g, 5.23 mmol) were added sequentially. The reaction mixture was stirred at room temperature for 1.5 h, quenched with water, and extracted with MTBE. The organic layer was washed with brine, dried over MgS04, and concentrated in vacuo. The residue was purified on a silica gel column with 0-50% EtOAc / cyclohexane to yield compound (11-1) (1.47 g, 4.42 mmol, 85% yield).

[0464] Step 2

[0465] Compound (11-1) (1.47 g, 4.42 mmol) was dissolved in THF (29.5 mL) and water (14.74 mL). LiOH-H20 (0.278 g, 6.63 mmol) was added at 0 °C. The reaction mixture was stirred vigorously at 0 °C for 30 min, quenched with 1 M HC1 (6.6 mL), and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified on a silica gel column with 0-15% MeOH / DCM to yield compound (11-2) (1.32 g).

[0466] Step 3

[0467] Compound (1-4) (50 mg, 0.187 mmol) and compound (11-2) (56.8 mg, 0.187 mmol) were dissolved in THF (1.6 mL) and DMF (0.3 mL), hunig's base (98 μΐ, 0.560 mmol) and HATU (56.8 mg, 0.149 mmol) were added. The reaction mixture was stirred at room temperature for 30 min, quenched with water and extracted with ethyl acetate. The organic layer was loaded on a silica gel column and eluted with 0-50% acetone / cyclohexane to yield compound (11-3) (75 mg, 0.145 mmol, 78% yield) as a mixture of two enantiomers.

[0468] Step 4

[0469] To a suspension of compound (11-3) (67 mg, 0.129 mmol) in DCM (1.3 mL) at 0 °C was added triethylamine (144 μΐ, 1.036 mmol) and TFAA (73.1 μΐ, 0.518 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 10 min. The reaction mixture was diluted with DCM and quenched with saturated NaHC03. The organic layer was loaded on a silica gel column and eluted with 0-50% EtOAc / cyclohexane to yield compound (11-4) (48 mg, 0.096 mmol, 74.2% yield) as a mixture of two enantiomers.

[0470] Step 5

[0471] Compound (11-4) (5 mg) was purified on a preparative HPLC with 20-85% MeCN / H20 with 0.1% formic acid to yield Example 11 (1.8 mg) and Example 12 (1.9 mg).

[0472] Example 11: 1H NMR (400 MHz, Acetone-d6) δ 10.47 (s, 1H), 9.56 (s, 1H), 7.74 (d, J = 8.1 Hz, 1H), 7.21 (d, J = 2.2 Hz, 1H), 7.10 - 6.89 (m, 5H), 6.85 (d, J = 7.7 Hz, 1H), 6.76 (td, J = 7.5, 1.0 Hz, 1H), 6.41 (dd, J = 7.3, 1.1 Hz, 1H), 5.03 (t, J = 8.2 Hz, 1H), 4.84 - 4.74 (m, 1H), 4.23 (d, J = 10.2 Hz, 1H), 3.91 (d, J = 10.3 Hz, 1H), 3.81 (s, 3H), 2.56 (td, J = 13.5, 8.2 Hz, 2H), 1.71 (ddd, J = 14.5, 9.9, 3.9 Hz, 2H), 1.58 (ddd, J = 13.8, 9.7, 4.9 Hz, 1H), 0.86 (dd, J = 11.9, 6.4 Hz, 6H). [M+Na] m / e 522.19.

[0473] Example 12 1 H NMR (400 MHz, Acetone-d6) δ 10.75 (s, 0.33H), 10.59 (s, 0.67H), 9.58 (s, 0.67H), 9.54 (s, 0.33H), 8.10 (d, J = 7.8 Hz, 0.33H), 7.90 (d, J = 8.7 Hz, 0.67H), 7.34 - 6.71 (m, 8H), 6.42 (m, 1H), 5.90 (t, J = 8.0 Hz, 0.33H), 5.06 (t, J = 8.3 Hz, 0.67H), 4.98 (ddd, J = 11.3, 7.7, 4.0 Hz, 0.33H), 4.83 (td, J = 9.1, 4.7 Hz, 0.67H), 4.00 (dd, J = 11.7, 1.4 Hz, 0.39H), 3.97 - 3.87 (m, 1.41H), 3.81 (m, 3H), 3.51 (d, J = 11.7 Hz, 0.39H), 2.65 - 2.49 (m, 1H), 1.91 (s, 2H), 1.71 - 1.51 (m, 2H), 0.96 - 0.90 (m, 2H), 0.75 (dd, J = 6.3, 4.1 Hz, 4H). [M+Na] m / e 522.19.

[0474] The following examples were prepared using similar protocols as described above.

[0475]

[0476]

[0477] Example 17

[0478]

[0479] Step 1

[0480] To a mixture of (2S)-2-amino-3-cyclobutylpropionic acid hydrochloride (0.359 g, 2 mmol) and NaOH (240 mg, 6.00 mmol) in toluene / water (4 mL / 4 mL) was added Cbz-Cl (0.314 ml, 2.200 mmol) at 0 °C. The mixture was stirred at room temperature for 2 h, the two layers were separated and the aqueous layer was washed with MBTE. The aqueous layer was then treated with 1 M HC1 solution to pH ~ 2. The resulting mixture was extracted with ethyl acetate. The collected organic layer was washed with brine, dried over Na2S04, filtered, and concentrated to give compound (17-1) (0.46 g, 1.659 mmol, 83% yield).

[0481] Step 2

[0482] To a mixture of compound (17-1) (104 mg, 0.374 mmol), compound (1-4) (80 mg, 0.299 mmol), and DIPEA (183 μΐ, 1.046 mmol) in DCM / DMF (1.0 / 0.5 mL) was added HATU (136 mg, 0.359 mmol) at room temperature. The reaction mixture was stirred at room temperature for 20 h, quenched with water, and extracted with ethyl acetate. The combined organic layers were washed with 1 N HC1, NaHC03sat. solution, brine, and dried over Na2S04and filtered. The filtrate was concentrated in vacuo. The residue was purified on a silica gel column to give compound (17-2) (98 mg, 0.200 mmol, 66.9% yield). [M-H] - , 489.16

[0483] Step 3

[0484] A suspension of (17-2) (25 mg, 0.051 mmol) and Pd-C (5.42 mg, 5.10 μmol) in MeOH (1 mL) was treated with 1 atm of hydrogen for 40 mins. The mixture was diluted with DCM, filtered over celite, washed with DCM, and concentrated in vacuo. The resulting product (17-3) was used directly in the next step. [M-H] - , 355.15

[0485] Step 4

[0486] To a suspension of 4-methoxy-lH-indole-2-carboxylic acid (15 mg, 0.077 mmol), compound (17-3) (18 mg, 0.051 mmol) and HATU (0.029 g, 0.077 mmol) in DCM (0.3 mL) was added a solution of DIPEA (0.031 ml, 0.179 mmol) in DMF (0.3 mL). The reaction mixture was stirred at room temperature for 1 h, quenched with water, and extracted with ethyl acetate. The combined organic layers were washed with IN HC1, NaHC03 saturated solution, brine, and dried over Na2S04and filtered. The residue was concentrated in vacuo. The residue was purified on a silica gel column to yield compound (17-4) (19 mg, 0.036 mmol, 70.3% yield). [M-H] - , 528.18

[0487] Step 5

[0488] To a mixture of compound (17-4) (19 mg, 0.036 mmol) and Et3N (60.0 μΐ, 0.431 mmol) in DCM (0.6 mL) was added TFAA (30.4 μΐ, 0.215 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was quenched with cold NaHC03 saturated solution and extracted with ethyl acetate. The combined organic layers were washed with IN HC1, NaHC03 saturated solution, brine, and dried over Na2S04and concentrated. The residue was purified on a silica gel column to yield Example 17 (12 mg, 0.023 mmol, 65.4% yield). [M-H] - 510.17; 1 H NMR (400 MHz, Methanol-d4) δ 7.26 (d, J = 0.9 Hz, 1H), 7.22 - 7.10 (m, 3H), 7.02 (d, J = 8.3 Hz, 1H), 6.99 - 6.89 (m, 2H), 6.53 (d, J = 7.7 Hz, 1H), 5.17 (t, J = 7.9 Hz, 1H), 4.71 (dd, J = 8.0, 6.4 Hz, 1H), 4.30 (d, J = 10.5 Hz, 1H), 4.08 (d, J = 10.5 Hz, 1H), 3.96 (s, 3H), 2.75 - 2.62 (m, 2H), 2.52 (hept, J = 7.7 Hz, 1H), 2.20 - 2.12 (m, 3H), 2.15 - 2.02 (m, 1H), 2.05 - 1.88 (m, 2H), 1.90 - 1.80 (m, 1H), 1.83 - 1.70 (m, 1H).

[0489]

[0490]

[0491]

[0492]

[0493]

[0494]

[0495]

[0496]

[0497] The following examples were prepared using similar protocols as described above.

[0498] Example 23

[0499]

[0500]

[0501] Step 1

[0502] To a solution of compound (1-2) (2.5 g, 7.22 mmol) in THF (24.06 mL) was added dropwise a solution of 2M LiBH4 in THF (10.83 mL, 21.65 mmol). The reaction mixture was stirred at room temperature for 2 h. And the bulk of THF was removed under vacuum. The reaction was carefully quenched to pH = 5-6 (~22 mL) with 1 N HCI and extracted with ethyl acetate (3 x 40 mL). The combined organic phase was washed with saturated sodium bicarbonate, brine, dried and concentrated. Purification of the residue on a silica gel column with 0-50% EtOAc / cyclohexane yielded the desired alcohol (23-1) (1.54 g, 67% yield).

[0503] Step 2

[0504] Compound (23-1) (0.5 g, 1.570 mmol) was dissolved in 4M HCI in dioxane (3.93 mL, 15.70 mmol), the reaction mixture was stirred at room temperature for 1 h and concentrated to dryness. Compound (23-2) was obtained as a yellow solid (492 mg, 80% yield). LC-MS, ES+: 218.85 [M+l].

[0505] Step 3

[0506] To a solution of compound (23-2) (960 mg, 3.13 mmol) and ((benzyloxy)carbonyl)-L-leucine (913 mg, 3.44 mmol) in dry DMF (15.64 mL) was added HATU (1546 mg, 4.07 mmol) and Hunig's base (1912 μΐ, 10.95 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h, diluted with EtOAc and washed with 10% citric acid, water, and brine. The organic layer was dried and concentrated. The residue was purified on a silica gel column with 0-40% EtOAc / cyclohexane to yield 1.2 g of compound (23-3). LC-MS, ES+: 466.19 [M+1].

[0507] Step 4

[0508] Compound (23-3) (800 mg, 1.718 mmol) was dissolved in MeOH (17 mL). Pd on carbon (10% loading) (40 mg, 0.038 mmol) was added. The reaction mixture was stirred under hydrogen atmosphere for 2.5 h and filtered through a pad of celite. The solvent was removed to yield crude (23-4) (543 mg, 1.638 mmol, 95% yield) which was used directly in the next step. [M+1] 332.20.

[0509] Step 5

[0510] Compound (23-4) (195 mg, 0.588 mmol) and 4-methoxy-1H-indole-2-carboxylic acid (118 mg, 0.618 mmol) were dissolved in CH2CI2(5.9 mL). Hunig's base (308 μΐ, 1.765 mmol) and HATU (235 mg, 0.618 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 30 min. The reaction mixture was quenched with water and extracted with DCM. The organic layer was loaded on a silica gel column and eluted with 0-50% acetone / cyclohexane to yield compound (23-5) (213 mg, 0.422 mmol, 71.7% yield).

[0511] Step 6

[0512] In a flame dried flask, acetic anhydride (422 μΐ, 4.46 mmol) was added to dry DMSO (3.10 mL) at room temperature. After stirring for 10 min, compound (23-5) (150 mg, 0.297 mmol) was added in one portion. The reaction mixture was stirred at room temperature for 6 h. The reaction mixture was cooled to 0 °C and diluted with water (~8 mL). The white precipitate was collected by filtration, rinsed with water, and dried under vacuum. Purification of the resulting precipitate on a silica gel column with 0-45% acetone / cyclohexane yielded example 23 as a colorless solid (112 mg, 75% yield). [M+H] + 503.16. 1 H NMR (500 MHz, DMSO-d6) δ 11.52 (d, J = 2.3 Hz, 1H), 10.66 (s, 1H), 9.52 (d, J = 2.1 Hz, 1H), 8.61 (d, J = 7.7 Hz, 1H), 7.40 - 7.32 (m, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.26 - 7.18 (m, 1H), 7.17 - 7.02 (m, 1H), 7.04 - 6.97 (m, 2H), 6.89 (d, J = 8.9 Hz, 1H), 6.52 (t, J = 8.4 Hz, 1H), 4.75 (s, 1H), 4.62 (td, J = 8.1, 7.1, 3.8 Hz, 1H), 4.11 (d, J = 10.5 Hz, 1H), 3.97 (d, J = 10.5 Hz, 1H), 3.89 (s, 3H), 3.88 (d, J = 7.4 Hz, 1H), 2.41 (dd, J = 13.0, 9.0 Hz, 1H), 2.21 (dd, J = 13.2, 6.2 Hz, 1H), 1.77 (m, 1H), 1.60 (m, 1H), 0.96 (d, J = 7.0 Hz, 3H), 0.89 (d, J = 7.0 Hz, 3H).

[0513] The following examples were prepared using similar protocols as described above.

[0514]

[0515]

[0516] Example 26

[0517]

[0518]

[0519] Step 1

[0520] Compound 23-4 (45 mg, 0.136 mmol) was dissolved in DCM (1.358 mL). DIPEA (48.5 μΐ, 0.272 mmol), 6-cyano-4-methoxy-lH-indole-2-carboxylic acid (32.3 mg, 0.149 mmol), and HATU (51.6 mg, 0.136 mmol) were added. The reaction mixture was stirred at room temperature for 1 h, quenched with water, and extracted with DCM. The organic layer was loaded on a silica gel column and eluted with 0-50% acetone / cyclohexane to yield compound 26-1 (22 mg, 0.042 mmol, 30.6% yield). [M-OH] + , 512.20.

[0521] Step 2

[0522] Acetic anhydride (78 μΐ, 0.831 mmol) was added to DMSO (0.415 mL) at room temperature. The reaction mixture was stirred at room temperature for 5 min, and the reaction mixture was transferred to a vial containing compound 26-1 (22 mg, 0.042 mmol). The resulting reaction mixture was stirred at room temperature for 6 h, quenched with water at 0 °C, and extracted with ethyl acetate. The organic layer was washed with water, brine, and concentrated. The residue was purified on a silica gel column with 0-50% acetone / cyclohexane to yield compound 26-2 (15 mg, 0.028 mmol, 68.4% yield). [M+H] + , 528.21.

[0523] Step 3

[0524] Compound 26-2 (15 mg, 0.028 mmol) was dissolved in 2-propanol. 1 M Hydroxylamine hydrochloride (56.9 μΐ, 0.057 mmol) in t-BuOH / H2O (1 : 1) was added. The reaction mixture was stirred at room temperature for 30 min, quenched with anhydrous NaHC03, and extracted with ethyl acetate. The organic layer was dried over Na2S04and concentrated in vacuo. The resulting crude compound 26-3 (14 mg, 0.026 mmol, 91% yield) was used directly in the next step. [M+H] + , 543.22

[0525] Step 4

[0526] To a vial containing compound 26-3 (14 mg, 0.026 mmol) was added MeCN (0.516 mL) and copper(II) acetate (1.406 mg, 7.74 μmol). The resulting reaction mixture was stirred at 70 °C for 2 h and concentrated in vacuo. The residue was purified on a silica gel column with 0-50% EtOAc / cyclohexane, followed by purification with prep-HPLC to yield Example 26 (2.8 mg, 5.34 μmol, 20.69% yield). [M+H] + , 525.22; 1 H NMR (400 MHz, Acetone-d6) δ 11.06 (s, 1H), 9.57 (s, 1H), 7.96 (d, J = 8.3 Hz, 1H), 7.45 (s, 1H), 7.32 (d, J = 1.9 Hz, 1H), 7.08 - 6.92 (m, 2H), 6.84 (d, J = 7.8 Hz, 1H), 6.77 - 6.68 (m, 2H), 5.03 (t, J = 8.3 Hz, 1H), 4.84 - 4.75 (m, 1H), 4.23 (d, J = 10.4 Hz, 1H), 3.91 (m, 5H), 2.58 (qd, J = 13.3, 8.4 Hz, 2H), 1.72 (m, 2H), 1.61 (m, 1H), 0.85 (m, 6H)

[0527] The following examples were prepared using similar protocols as described above.

[0528]

[0529]

[0530]

[0531] Example 29

[0532]

[0533] To a solution of Example 23 (45 mg, 0.090 mmol) in EtOH (2 mL) and water (0.2 mL) was added sodium persulfate (9.32 mg, 0.090 mmol). The reaction mixture was stirred at room temperature for 4 h and then concentrated. DCM was added to the residue to obtain a white solid precipitate. The collected solid was washed with acetone and dried to yield Example 29 as a white solid. [M-Na] - 583.0. 1H NMR (500 MHz, DMSO-d6) δ 11.42 (s, 1H), 10.57 (d, J = 7.9 Hz, 1H), 9.88 (s, 1H), 8.47 (d, J = 8.2 Hz, 1H), 7.35 - 7.31 (m, 1H), 7.14 - 7.05 (m, 2H), 7.02 - 6.96 (m, 1H), 6.86 (ddt, J = 24.0, 15.0, 8.1 Hz, 3H), 6.50 (d, J = 7.7 Hz, 1H), 5.65 (d, J = 5.5 Hz, 1H), 4.83 - 4.78 (m, 1H), 4.70 (t, J = 9.3 Hz, 2H), 3.96 (d, J = 9.3 Hz, 1H), 3.90 (s, 3H), 3.61 (d, J = 9.8 Hz, 1H), 2.79 (dd, J = 13.1, 9.8 Hz, 1H), 1.81 - 1.67 (m, 3H), 0.99 (td, J = 15.4, 7.0 Hz, 1H), 0.90 (d, J = 6.4 Hz, 3H), 0.85 (d, J = 6.1 Hz, 3H).

[0534] The following examples were prepared using similar methods described above.

[0535]

[0536] Example 31

[0537]

[0538] Step 1

[0539] To Example 23 (18 mg, 0.036 mmol) was added acetic acid (2.4 μΐ, 0.041 mmol) and isocyanocyclopropane (2.64 mg, 0.039 mmol) in DCM (0.20 mL) at 0 °C. The reaction mixture was stirred at a temperature range from 0 °C to room temperature for 5 hours. The reaction mixture was concentrated to dryness and redissolved in MeOH (0.35 mL). 0.5 M K2CO3 in water (179 μΐ, 0.090 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The MeOH was removed in vacuo and the aqueous layer was extracted with ethyl acetate (3x). The combined organic layers were washed with water and brine, dried and concentrated. The resulting crude product (31-1) was used directly in the next step. [M+1], 588.2.

[0540] Step 2

[0541] To a solution of compound (31-1) in DCM (0.360 mL) was added Dess-Martin Oxidizing reagent (0.023 g, 0.054 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2.5 h. The reaction mixture was diluted with DCM, quenched with 10% Na2S2O3, and washed with 5% NaHCO3. The collected organic layer was washed with water and brine, dried, and concentrated. The residue was purified on a silica gel column with 0-60% acetone / cyclohexane to yield Example 31 (6.5 mg).[M-1] - 584.07. 1 H NMR (400 MHz, Acetone-de) δ 10.62 (s, 1H), 9.67 (s, 1H), 7.90 (d, J = 4.9 Hz, 1H), 7.78 (d, J = 8.2 Hz, 1H), 7.31 (dd, J = 2.3, 0.8 Hz, 1H), 7.29 - 7.10 (m, 4H), 7.14 - 6.95 (m, 2H), 6.92 (td, J = 7.6, 1.1 Hz, 1H), 6.53 (dd, J = 7.2, 1.2 Hz, 1H), 5.69 - 5.54 (m, 1H), 4.93 (td, J = 8.4, 6.0 Hz, 1H), 4.34 (d, J = 9.9 Hz, 1H), 4.02 (d, J = 9.9 Hz, 1H), 3.94 (s, 3H), 4.00 - 3.86 (m, 1H), 2.92 - 2.78 (m, 1H), 2.52 - 2.38 (m, 2H), 1.89 (dt, J = 12.9, 6.5 Hz, 1H), 1.72 (ddd, J = 8.1, 5.7, 2.3 Hz, 2H), 1.13 - 0.93 (m, 6H), 0.83 - 0.65 (m, 4H).

[0542] The following examples were prepared using similar protocols as described above.

[0543]

[0544]

[0545]

[0546]

[0547] Example 37

[0548]

[0549] To a mixture of Example 23 (105 mg, 0.209 mmol) in t-butanol (2.79 mL) was added 2-methyl-2-butene, 2M in THF (2.09 mL, 4.18 mmol) at room temperature to give a clear solution. A solution of sodium chlorite (236 mg, 2.089 mmol) and sodium monobasic phosphate (251 mg, 2.089 mmol) in water (1.39 mL) was added dropwise over 10 min. The reaction mixture was stirred at room temperature for 1 h, concentrated to remove most of the volatiles. The resulting mixture was diluted with ethyl acetate, washed with water, brine, dried and concentrated. The residue was purified on silica gel chromatography with 0-10% MeOH / DCM to give Example 37 (40 mg, 36% yield). LC-MS, ES+: 516.94 [M-H] - - .

[0550] Example 38

[0551]

[0552] A solution of Example 37 (18 mg, 0.035 mmol), cyclopropane sulfonamide (8.41 mg, 0.069 mmol), EDCI (7.2 mg, 0.038 mmol) and DMAP (4.59 mg, 0.038 mmol) in dry DCM was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM, washed with brine, dried, and concentrated. The resulting residue was chromatographed on silica gel column with 0 to 50% acetone / cyclohexane to give Example 38 (3.5 mg, 16% yield) as a white solid. LC-MS, ES+: 619.80 [M-H] - .

[0553] Example 157

[0554]

[0555]

[0556] ​Step 1 : To a suspension of methyltriphenylphosphonium bromide (479 mg, 1.34 mmol) (evaporated twice with dry toluene prior to use) in THF (4.2 mL) was added potassium tert-butoxide (1 M in THF, 1.26 mL, 1.26 mmol) at 0 °C. The mixture turned into a yellowish paste. After stirring at 0 °C for 0.5 h, a solution of 157-2 (200 mg, 0.419 mmol) in THF (1.0 mL) was added dropwise at 0 °C. The resulting yellow paste was stirred at 0 °C for 1 h. The reaction was quenched by the addition of excess saturated NH4CI solution. The mixture was diluted with ethyl acetate and water. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on silica gel chromatography with 0-50% ethyl acetate in cyclohexane to give 157-3 (160 mg, 80% yield). LC-MS, ES + : 476.10 [M+1].

[0557] Step 2: To a mixture of 157-3 (112 mg, 0.235 mmol) and NMO (83 mg, 0.706 mmol) in acetone (2.10 mL) / water (0.24 mL) was added osmium tetroxide 2.5% in tBuOH (443 μί, 0.035 mmol) at room temperature. After stirring at room temperature for about 3 h, the resulting reaction mixture was quenched with saturated Na2S03solution and then extracted with ethyl acetate (2x). The combined organic phase was washed with brine, dried over sodium sulfate, and concentrated. The crude product 157-4 (118 mg, 98% yield) was used directly in the next step without further purification. LC-MS, ES - : 508.1 [M-H].

[0558] Step 3: To a solution of 157-4 (118 mg, 0.232 mmol) in CH2CI2(2.32 mL) and 1 H-imidazole (23.65 mg, 0.347 mmol) was added tert-butyldimethylsilyl chloride (36.6 mg, 0.243 mmol) at 0 °C. After stirring at room temperature for about 2 hrs, the resulting reaction mixture was quenched with saturated sodium sulfite and extracted with ethyl acetate (2x). The combined organic phase was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified on silica gel chromatography with 0-40% ethyl acetate in cyclohexane to give 157-5 (126 mg, 87% yield). LC-MS, ES + : 624.28 [M+1].

[0559] Step 4: A mixture of 157-4 (126 mg, 0.202 mmol) and 10% Pd-C (21.49 mg, 0.020 mmol) in MeOH (2.0 mL) was stirred under an atmosphere of hydrogen at room temperature. After ~1 h, the reaction mixture was filtered through celite, washed with methanol, and concentrated to yield crude 157-5 (99 mg, 100% yield) which was used directly in the next step. LC-MS, ES + : 490.5 [M+1].

[0560] Step 5: To a solution of 157-5 (99 mg, 0.202 mmol) and 4,6-difluoro-lH-indole-2- carboxylic acid (39.9 mg, 0.202 mmol) in DMF (0.81 mL) was added 4- methylmorpholine (66.7 μί, 0.606 mmol) followed by HATU (85 mg, 0.222 mmol) at 0 °C. The resulting reaction mixture was then stirred at room temperature for 3-4 h. Check: Dilute the reaction mixture with ethyl acetate, wash with water (2x), brine, dry over sodium sulfate, and concentrate. Purify the residue on a silica gel chromatography column with 0-40% ethyl acetate in cyclohexane to yield 157-6 (105 mg, 0.157 mmol, 78% yield). LC-MS, ES + : 669.22 [M+1].

[0561] Step 6: To a solution of 157-6 (104 mg, 0.155 mmol) in DCM (1.56 mL) was added Dess-Martin Oxidizer (198 mg, 0.46 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 4-5 h until TLC (acetone / cyclohexane 1 / 3) showed all sm were consumed. Check: Dilute the reaction mixture with DCM, quench the reaction mixture with 10% Na2S2O3 and 5% NaHCO3. Separate the organic layer, wash with water, brine, dry over sodium sulfate, and concentrate. Purify the residue on a silica gel chromatography column with 0-40% acetone / cyclohexane to yield 157-7 (50 mg, 0.075 mmol, 48.2% yield). LC-MS, ES - : 665.0 [M-H].

[0562] Step 7: To a suspension of 157-7 (42 mg, 0.063 mmol) in MeOH (0.63 mL) was added concentrated high concentration aqueous hydrochloric acid (31.5 μί, 0.378 mmol) at room temperature. The reaction mixture was stirred at room temperature for about 15 min and concentrated under vacuum until dryness. The residue was diluted with ethyl acetate, washed with saturated sodium bicarbonate, brine, dried over sodium sulfate, and concentrated. The residue was purified on silica gel chromatography with 0-50% acetone in cyclohexane to yield Example 157- (26 mg, 0.047 mmol, 74.7% yield). LC-MS, ES - : 550.90 [M-H].1H NMR (400 MHz, Acetone-d6) δ 10.83 (s, 1H), 9.63 (s, 1H), 7.07 (dd, J = 7.7, 4.5 Hz, 3H), 6.97 - 6.83 (m, 3H), 6.74 (td, J = 10.3, 2.1 Hz, 1H), 5.55 (t, J = 7.6 Hz, 1H), 5.12 (dd, J = 9.6, 8.1 Hz, 1H), 4.56 (dd, J = 18.6, 5.7 Hz, 1H), 4.42 (dd, J = 18.6, 5.8 Hz, 1H), 4.26 (d, J = 10.3 Hz, 1H), 4.09 (t, J = 5.7 Hz, 1H), 3.93 (d, J = 10.2 Hz, 1H), 3.46 (s, 3H), 2.44 (ddd, J = 12.7, 8.1, 1.3 Hz, 1H), 2.36 (dd, J = 12.7, 9.6 Hz, 1H), 1.81 (tq, J = 14.0, 7.4, 6.7 Hz, 2H), 1.63 (ddd, J = 14.3, 12.9, 6.7 Hz, 1H), 1.02 (d, J = 6.6 Hz, 3H), 0.96 (d, J = 6.5 Hz, 3H).

[0563] Example 158

[0564]

[0565] Example 158 was prepared using a similar protocol as described above. [M-1] 569.0; 1H NMR (400 MHz, Acetone-d6) δ 11.25 (s, 1H), 9.63 (s, 1H), 7.13 - 7.02 (m, 2H), 6.96 - 6.81 (m, 4H), 5.54 (t, J = 7.6 Hz, 1H), 5.14 (dd, J = 9.7, 8.1 Hz, 1H), 4.57 (dd, J = 18.7, 5.7 Hz, 1H), 4.42 (dd, J = 18.7, 5.8 Hz, 1H), 4.23 (d, J = 10.5 Hz, 1H), 4.11 (t, J = 5.7 Hz, 1H), 3.95 (d, J = 10.3 Hz, 1H), 3.45 (s, 3H), 2.45 (dd, J = 12.7, 8.1 Hz, 1H), 2.36 (dd, J = 12.7, 9.7 Hz, 1H), 1.90 - 1.74 (m, 2H), 1.67 (dp, J = 13.6, 6.7 Hz, 1H), 1.03 (d, J = 6.6 Hz, 3H), 0.97 (d, J = 6.5 Hz, 3H).

[0566] Example 159

[0567]

[0568] To a solution of Example 158 (11.6 mg, 0.020 mmol) in THF (0.25 mL) at 0 °C was added lithium chloride (11.20 mg, 0.264 mmol) and Hunig's base (10.6 μL, 0.061 mmol). Then, methanesulfonyl chloride (3.78 μL, 0.048 mmol) was added. The reaction mixture was stirred at 0 °C to room temperature for about 6 hours. Check: Quench the reaction mixture with saturated NH4Cl, extract with ethyl acetate. Separate the organic layer, wash with brine, dry over sodium sulfate, and concentrate. Purify the residue on a silica gel chromatography column with 0-30% acetone in cyclohexane to yield Example 159 (4.5 mg, 37.6% yield). LC-MS, ES -586.86 [M-H]; 1H NMR (400 MHz, Acetone-d6) δ 11.22 (s, 1H), 9.64 (s, 1H), 7.18 - 7.06 (m, 2H), 7.00 - 6.85 (m, 3H), 5.55 (t, J = 7.7 Hz, 1H), 5.13 (t, J = 8.5 Hz, 1H), 4.81 (d, J = 16.5 Hz, 1H), 4.65 (d, J = 16.5 Hz, 1H), 4.23 (d, J = 10.6 Hz, 1H), 4.00 (d, J = 10.3 Hz, 1H), 3.44 (s, 3H), 2.55 - 2.38 (m, 2H), 1.92 - 1.74 (m, 2H), 1.65 (dt, J = 13.9, 6.7 Hz, 1H), 1.00 (dd, J = 22.0, 6.6 Hz, 6H).

[0569] Example 39

[0570]

[0571] Step 1

[0572] Compound (1-4) (300 mg, 1.121 mmol) and N-((benzyloxy)carbonyl)-N- methyl-L-leucine (344 mg, 1.233 mmol) were taken in CH2Cl2(5 ml) and DMF (1 ml). To this was added 4-methylmorpholine (246 μl, 2.241 mmol) and HATU (469 mg, 1.233 mmol). The reaction mixture was stirred at room temperature for 1 h, diluted with DCM (30 mL) and washed with saturated NaHC03. The collected organic layer was washed with 1M HC1 and brine, filtered over Na2S04and concentrated in vacuum. The residue was purified on silica gel column with 0-100% acetone / cyclohexane to yield compound (39-1) (417 mg, 0.847 mmol, 76% yield). [M-1] - , 491.02.

[0573] Step 2

[0574] To a suspension of (39-1) (28 mg, 0.057 mmol) in DCM (0.6 mL) at 0 °C was added Et3N (79 μΐ, 0.568 mmol) and TFAA (40.1 μΐ, 0.284 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction was quenched with cold NaHC03solution and extracted with ethyl acetate. The organic layer was washed with water, 1 N HC1, saturated NaHC03and brine, dried over Na2S04, filtered, and concentrated. The residue was purified on a silica gel column to yield Example 39 (24 mg, 0.051 mmol, 89% yield). [M-H] - 473.17. 1 H NMR (400 MHz, Methanol-d4) δ 7.35 - 7.15 (m, 5H), 7.07 - 6.88 (m, 4H), 5.21 - 4.88 (m, 2H), 4.77 (dd, J = 12.1, 3.8 Hz, 1H), 4.19 - 4.07 (m, 1H), 3.92 (d, J = 10.7 Hz, 1H), 3.71 (p, J = 10.8 Hz, 1H), 2.93 (d, J = 4.8 Hz, 3H), 2.75 - 2.57 (m, 2H), 1.79 (ddt, J = 14.4, 9.2, 5.0 Hz, 1H), 1.67 (dq, J = 14.8, 7.2, 6.6 Hz, 1H), 1.56 - 1.47 (m, 1H), 1.05 - 0.88 (m, 6H).

[0575] The following examples were prepared using a similar protocol as described above.

[0576]

[0577]

[0578] Example 42

[0579]

[0580] Step 1

[0581] Compound (39-1) (1323 mg, 2.69 mmol) was dissolved in MeOH (30 ml). 10% Pd-C (143 mg, 0.134 mmol) was added. The reaction mixture was stirred under hydrogen gas (balloon) for 1 hour and filtered over a celite pad. The filtrate was concentrated in vacuo to yield compound (42-1). [M+H] + 359.2

[0582] Step 2

[0583] To a suspension of 4-methoxy-lH-indole-2-carboxylic acid (0.111 g, 0.583 mmol), compound (42-1) (0.182 g, 0.507 mmol) and HATU (0.212 g, 0.558 mmol) in DCM (0.3 mL) was added a solution of DIPEA (0.266 ml, 1.521 mmol) in DMF (0.35 mL). The reaction mixture was stirred at room temperature for 1 h, quenched with water, and extracted with ethyl acetate. The organic layer was washed with 1 N HC1, saturated NaHC03and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column to yield compound (42-2) (160 mg, 0.301 mmol, 59.4% yield). [M-H] - 530.18.

[0584] Step 3

[0585] Compound (42-2) (150 mg, 0.282 mmol) was dissolved in CH2Cl2(1.9 ml). Et3N (0.32 mL, 2.26 mmol) and TFAA (0.16 mL, 1.13 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 20 min, extracted with aqueous NaHC03solution, and extracted with DCM (2x). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. The residue was purified on a silica gel column with 0-40% acetone / cyclohexane to yield Example 42 (114 mg, 0.222 mmol, 79% yield). [M-H] - 512.18; 1 H NMR (400 MHz, Methanol-d4) δ 7.15 (t, J = 8.0 Hz, 1H), 7.05 (t, J = 7.8 Hz, 1H), 7.01 - 6.94 (m, 2H), 6.90 (s, 1H), 6.85 (dd, J = 15.4, 7.7 Hz, 2H), 6.52 (d, J = 7.7 Hz, 1H), 5.53 (br s, 1H), 5.19 (t, J = 8.0 Hz, 1H), 4.21 (d, J = 11.0 Hz, 1H), 3.99 (d, J = 11.0 Hz, 1H), 3.96 (s, 3H), 3.40 (s, 3H), 2.75 - 2.60 (m, 2H), 1.96 - 1.76 (m, 2H), 1.63 (ddt, J = 14.6, 13.0, 6.6 Hz, 1H), 1.47 (s, 1H), 1.26 (t, J = 7.1 Hz, 1H), 1.01 (m, 6H).

[0586] The following examples were prepared using similar protocols as described above.

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610]

[0611]

[0612]

[0613]

[0614]

[0615]

[0616]

[0617]

[0618]

[0619]

[0620]

[0621]

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638]

[0639]

[0640]

[0641]

[0642]

[0643]

[0644]

[0645]

[0646]

[0647]

[0648]

[0649]

[0650]

[0651]

[0652]

[0653] Example 367

[0654]

[0655] To a solution of Example 366 (17 mg, 0.033 mmol) in DCM (2 mL) was added TFA (0.03 mL) and the resulting reaction mixture was stirred at room temperature for 4 hours. The solvent was removed to yield the title compound Example 367 (17 mg, 100%). [M+1] 412.47.

[0656] Example 368

[0657]

[0658] To a solution of Example 367 (61 mg, 0.12 mmol) and 2,4-difluorobenzoic acid (19 mg, 0.12 mmol) in DMF (2 mL) was added HATU (46 mg, 0.12 mmol) and DIPEA (0.04 mL, 0.36 mmol). The resulting reaction mixture was stirred at room temperature for 4 h and concentrated. The crude product was purified on a silica gel column to give Example 368 (17 mg, 21%). [M-1] 550.36

[0659] Example 89

[0660]

[0661]

[0662] Step 1

[0663] To a mixture of (S)-2-(((benzyloxy)carbonyl)amino)-3-cyclobutylpropanoic acid (2.68 g, 9.66 mmol) and Mel (4.83 mL, 77 mmol) in THF (30 mL) was added NaH (1.16 g, 29 mmol) in portions at 0 °C. The resulting reaction mixture was stirred at room temperature for 2 days, quenched with ice water and washed with MBTE (2x). The aqueous layer was acidified to pH ~ 2 with 1 N HC1 and extracted with ethyl acetate. The collected organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated to give the desired compound (89-1) (2.54 g, 90% yield). ESI-MS m / z = 290.12 [M-H] - .

[0664] Step 2

[0665] To a solution of compound (1-4) (2.33 g, 6.96 mmol), compound (89-1) (2.54 g, 8.70 mmol) and 4-methylmorpholine (3.06 mL, 27.9 mmol) in DCM / DMF (5 / 5 mL) was added HATU (2.78 g, 7.31 mmol). The reaction mixture was stirred at room temperature for 2 h, quenched with water, and extracted with ethyl acetate. The collected organic layer was washed with water, 1 N HC1, saturated NaHC03and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column to give compound (89-2) (3.16 g, 90% yield). ESI-MS m / z = 503.19 [M-H] - .

[0666] Step 3

[0667] To a mixture of compound (89-2) (45 mg, 0.089 mmol) and Et3N (99 μΐ, 0.713 mmol) in DCM (1 mL) was added TFAA (50.4 μΐ, 0.357 mmol) dropwise at 0 °C. The resulting reaction mixture was stirred at room temperature for 30 min, quenched with cold saturated aqueous NaHC03solution, and extracted with ethyl acetate. The collected organic layers were washed with water, 1 N HC1, saturated NaHC03, and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column to yield Example 89 (23 mg, 53% yield). ESI-MS m / z = 485.19 [M-H] - .

[0668] The following examples were prepared in a similar manner as described above.

[0669]

[0670] Example 91

[0671]

[0672]

[0673] Step 1

[0674] To a solution of compound (89-2) (65 mg, 0.13 mmol) and Pd-C (13.7 mg, 0.013 mmol) in MeOH (1 mL) was passed hydrogen using a hydrogen balloon. After 1 h, the mixture was diluted with DCM, filtered over celite, and concentrated to yield compound (91-1) (48 mg, 100%). ESI-MS m / z = 369.19 [M-H] - .

[0675] Step 2

[0676] To a mixture of compound (91-1) (0.032 g, 0.086 mmol), 4,6-difluoro-lH-indole-2- carboxylic acid (0.021 g, 0.108 mmol), DIPEA (0.045 mL, 0.258 mmol) in DCM / DMF (0.5 / 0.5 mL) was added HATU (39 mg, 0.103 mmol) at room temperature. The resulting reaction mixture was stirred at room temperature for 20 h, quenched with water, and extracted with ethyl acetate. The collected organic layers were washed with water and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column to yield compound (91-2) (34 mg, 72% yield). ESI-MS m / z = 548.21 [M-H] - .

[0677] Step 3

[0678] To a mixture of compound (91-2) (34 mg, 0.062 mmol) and Et3N (86 μΐ, 0.619 mmol) in DCM (1 mL) was added TFAA (44 μΐ, 0.31 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 30 min, quenched with cold saturated aqueous NaHC03solution, and extracted with ethyl acetate. The collected organic layers were washed with 1 N HC1, saturated NaHC03, brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column with 0-40% acetone / cyclohexane to yield Example 91 (17 mg, 52% yield). ESI-MS m / z = 530.20 [M-H] - . 1 H NMR (400 MHz, Acetone-de) δ 10.65 (s, 1H), 9.51 (s, 1H), 6.97 - 6.83 (m, 3H), 6.81 - 6.72 (m, 2H), 6.67 (t, J = 7.6 Hz, 1H), 6.60 (td, J = 10.3, 2.1 Hz, 1H), 5.28 (t, J = 7.4 Hz, 1H), 5.05 (t, J = 8.2 Hz, 1H), 4.08 (d, J = 10.7 Hz, 1H), 3.82 (d, J = 10.6 Hz, 1H), 3.28 (s, 3H), 2.69 (s, 1H), 2.67 - 2.48 (m, 2H), 2.22 (hept, J = 7.7 Hz, 1H), 1.89 (d, J = 7.4 Hz, 3H), 1.74 - 1.53 (m, 4H).

[0679] The following examples were prepared using similar protocols as described above.

[0680]

[0681]

[0682]

[0683]

[0684]

[0685]

[0686]

[0687]

[0688]

[0689]

[0690]

[0691] Example 96

[0692]

[0693] Step 1

[0694] To a solution of ((benzyloxy)carbonyl)-L-leucine (1.56 g, 5.88 mmol) and 3- iodo prop-1 -ene (0.807 mL, 8.82 mmol) in THF (30 mL) was added NaH (0.706 g, 17.64 mmol) in portions at 0 °C. The reaction mixture was stirred at room temperature for 4 days, quenched with ice water and washed with MBTE twice. The aqueous layer was acidified to PH ~ 2 with 1 N HCI and extracted with ethyl acetate. The collected organic layer was washed with brine, dried over Na2S04, filtered and concentrated to yield compound (96-1) (1.15 g, 64.0% yield). ESI-MS m / z = 304.12 [M-H] - .

[0695] Step 2

[0696] To a solution of compound (1-4) (221 mg, 0.826 mmol), compound (96-1) (265 mg, 0.868 mmol) and DIPEA (577 μΐ, 3.31 mmol) in DCM / DMF (0.8 / 0.8 mL) was added HATU (314 mg, 0.826 mmol). The resulting reaction mixture was stirred at room temperature for 16 h, quenched with water and extracted with ethyl acetate. The organic layer was washed with water, 1 N HCI, saturated NaHC03and brine, dried over sodium sulfate, filtered and concentrated. The resulting residue was purified on silica gel chromatography with 0-10% MeOH / DCM to yield compound (96-2) (262 mg, 61.1% yield). ESI-MS m / z = 517.20 [M-H] - .

[0697] Step 3

[0698] To a solution of compound (96-2) (22 mg, 0.042 mmol) and Et3N (59.1 μΐ,, 0.424 mmol) in DCM (1 mL) was added TFAA (30.0 μΐ,, 0.212 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 30 min, quenched with cold saturated aqueous NaHC03solution, and extracted with ethyl acetate. The organic layer was washed with water, 1 N HC1, saturated NaHC03, and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column with 0-50% acetone / cyclohexane to yield Example 96 (20 mg, 94% yield). ESI-MS m / z = 499.20 [M-H] - .

[0699] Example 97

[0700]

[0701] Step 1

[0702] A mixture of compound (96-2) (105 mg, 0.202 mmol) and Pd-C (21.55 mg, 0.020 mmol) in MeOH (3 mL) was stirred under hydrogen atmosphere using a hydrogen balloon. After 1 h, the mixture was diluted with DCM, filtered over celite, and concentrated to yield compound (97-1) (79 mg, 100%). ESI-MS m / z = 385.19 [M-H] - .

[0703] Step 2

[0704] To a mixture of compound (97-1) (0.039 g, 0.10 mmol) in DCM / DMF (0.5 / 0.5 mL) and Et3N (0.098 mL, 0.70 mmol) was added Cbz-Cl (0.042 mL, 0.30 mmol). The reaction mixture was stirred at room temperature for 16 h, quenched with liquid NH3, and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over N2S04, filtered, and concentrated. The resulting residue was purified on a silica gel chromatography using 0-10% MeOH / DCM to yield compound (97-2) (10 mg, 19% yield). ESI-MS m / z = 519.22 [M-H] - .

[0705] Step 3

[0706] To a mixture of compound (97-2) (10 mg, 0.019 mmol) and Et3N (53.5 μΐ, 0.384 mmol) in DCM (0.5 mL) was added TFAA (27.1 μΐ, 0.192 mmol) at 0 °C. Quench with cold saturated aqueous NaHC03solution and extract with ethyl acetate. Wash the organic layer with 1 N HC1, saturated NaHC03solution and brine, dry over sodium sulfate, filter, and concentrate. Purify the resulting residue on silica gel chromatography using 0-50% acetone / cyclohexane to yield Example 97 (7.0 mg, 72.5% yield) ESI-MS m / z = 501.22 [M-H] - .

[0707] Example 98

[0708]

[0709] Step 1

[0710] Stir a mixture of 4-fluoro-lH-indole-2-carboxylic acid (0.054 g, 0.30 mmol) and 1- chloro-N,N,2-trimethylprop-l-en-l-amine (0.044 mL, 0.330 mmol) in DCM (1 mL) at room temperature for 1 h. Add the resulting mixture to a solution of compound (97-1) and Et3N (0.108 mL, 0.85 mmol) in DCM / DMF (0.5 / 0.5 mL). Stir the resulting reaction mixture at room temperature for 20 h, quench with liquid NH3and extract with ethyl acetate. Wash the organic layer with water and brine, dry over sodium sulfate, filter, and concentrate. Purify the resulting residue on silica gel chromatography using 0-10% MeOH / DCM to yield compound (98-1) (40 mg, 69% yield). ESI-MS m / z = 546.23 [M-H] - .

[0711] Step 2

[0712] To a solution of compound (98-1) (40 mg, 0.073 mmol) and Et3N (10.18 μΐ, 0.073 mmol) in DCM (1 mL) was added TFAA (10.32 μΐ, 0.073 mmol) at 0 °C. Stir the reaction mixture at room temperature for 30 min, quench with cold saturated NaHC03solution and extract with ethyl acetate. Wash the organic layer with 1 N HC1, saturated NaHC03and brine, dry over sodium sulfate, filter, and concentrate. Purify the resulting residue on silica gel chromatography using 0-50% acetone / cyclohexane to yield Example 98 (35 mg, 90% yield). ESI-MS m / z = 528.20 [M-H] -

[0713] The following examples were prepared in a similar manner as described above.

[0714]

[0715] Example 100

[0716]

[0717]

[0718] Synthesis of (S)-2-(((benzyloxy)carbonyl)(methyl)amino)-5-methylhexanoic acid

[0719] Step 1:

[0720] To a solution of (S)-2-amino-5-methylhexanoic acid (0.9 g, 6.20 mmol) in toluene / water (12.4 mL / 3 mL) at 0 °C was added 2 N NaOH (9.30 mL, 18.59 mmol) followed by Cbz-Cl (0.973 mL, 6.82 mmol). After stirring at room temperature for 2 h, the two layers were separated and the aqueous layer was washed with MBTE (2x) and the resulting reaction solution was acidified to pH ~ 2 with 1 N HC1 solution at 0 °C. The reaction mixture was extracted with ethyl acetate (3x). The combined organic phase was washed with brine, dried over sodium sulfate and concentrated to yield (S)-2-(((benzyloxy)carbonyl)amino)-5-methylhexanoic acid (1.42 g, 5.08 mmol, 82% yield) which was used in the next step without further purification. LC-MS, ES-: 277.77 [M-1].

[0721] Step 2:

[0722] To a mixture of (S)-2-(((benzyloxy)carbonyl)amino)-5-methylhexanoic acid (660 mg, 2.363 mmol) and paraformaldehyde (426 mg, 14.18 mmol) in dry acetonitrile (11.8 mL) was added 4-methylbenzylsulfonic acid hydrate (44.9 mg, 0.236 mmol). The resulting mixture was heated at 130 °C for 10 min in a microwave. After cooling to room temperature, the mixture was filtered over celite, concentrated and trapped with DCM to yield the crude product benzyl (S)-4-isopentyl-5-oxooxazolidine-3-carboxylate as a viscous oil which was used in the next step without further purification.

[0723] Step 3:

[0724] ​To the crude benzyl (S)-4-isopentyl-5-oxooxazolidine-3-carboxylate obtained in the previous step was added DCM (24 mL), triethylsilane (1.89 mL, 11.81 mmol), and 2,2,2-trifluoroacetic acid (7.28 mL, 95 mmol). The reaction mixture was stirred at room temperature for 2 h, concentrated and taken up in DCM (3x). The residue was basified with 1 N NaOH to pH ~ 10 at 0 °C and washed with EtOAc (lx) and MBTE (lx). The aqueous layer was acidified with 1 N HC1 to pH ~ 2 and extracted with ethyl acetate (2x). The combined organic phase was washed with brine, dried, and concentrated to yield compound (S)-2-(((benzyloxy)carbonyl)(methyl)amino)-5-methylhexanoic acid (715 mg, 92% yield over 2 steps).1H NMR (400 MHz, DMSO-d6) δ 12.56 (s, 1H), 7.41 - 7.27 (m, 5H), 5.17 - 5.00 (m, 2H), 4.48 (ddd, J = 27.4, 11.1, 4.7 Hz, 1H), 2.81 (s, 2H, N-Me rotamers), 2.78 (s, 1H, N-Me rotamers), 1.84 (tq, J = 9.6, 4.6, 4.1 Hz, 1H), 1.70 (ddd, J = 14.4, 9.6, 4.5 Hz, 1H), 1.52 (dt, J = 12.8, 6.5 Hz, 1H), 1.21 - 0.99 (m, 2H), 0.84 (dd, J = 9.2, 6.6 Hz, 6H).

[0725]

[0726] Synthesis Example 100

[0727] Step 1:

[0728] To a mixture of (S)-2-(((benzyloxy)carbonyl)(methyl)amino)-5- methylhexanoic acid (300 mg, 1.023 mmol) and (1-4) (261 mg, 0.974 mmol) in dry CH2Cl2(2.96 mL) was added DIPEA (510 μΐ, 2.92 mmol) and HATU (481 mg, 1.266 mmol) at 0 °C. The resulting reaction mixture was stirred at room temperature for 2 h. The mixture was diluted with DCM, washed with water (2x), brine, dried, and concentrated. The residue was purified on a silica gel column with 0-10% MeOH / DCM to give benzyl ((S)-1-((3R,5'S)-5'-carbamoyl-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-5-methyl-1- oxohexan-2-yl)(methyl)carbamate (100-1) (189 mg, 38% yield). LC-MS, ES-: 505.0 [M-1].

[0729] Step 2

[0730] To a mixture of compound (100-1) (31 mg, 0.061 mmol) and Et3N (85 μΐ, 0.612 mmol) in dry DCM (0.8 mL) was added TFAA (43.2 μΐ, 0.306 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 1 h, diluted with DCM, washed with saturated NaHC03, water, brine, dried and concentrated. The resulting residue was purified on silica gel chromatography with 0-40% acetone / cyclohexane to yield Example 100 (25 mg, 84% yield). LC-MS, ES + : 488.96 [M+1].

[0731] The following examples were prepared using similar protocols as described above.

[0732]

[0733] Example 104

[0734]

[0735] Step 1:

[0736] A mixture of compound (100-1) (152 mg, 0.300 mmol) and 10% Pd-C (31.9 mg, 0.030 mmol) in MeOH (3.00 mL) was stirred at room temperature under a hydrogen atmosphere. After 1 h, the mixture was filtered over celite, rinsed with MeOH, and concentrated to yield crude (3R,5'S)-1'-((S)-5-methyl-2-(methylamino)hexanoyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (104-1) (112 mg, 0.301 mmol, 100% yield), which was used directly in the next step. LC-MS, ES+: 372.99 [M+H] + .

[0737] Step 2:

[0738] To a mixture of compound (104-1) (85 mg, 0.228 mmol) and 4,6-difluoro-1H-indole-2- carboxylic acid (47.2 mg, 0.240 mmol) in dry DMF (1.14 mL) was added Hunig's base (122 μL, 0.685 mmol) and HATU (113 mg, 0.297 mmol) at 0 °C. The resulting reaction mixture was then stirred at room temperature for 1 h, diluted with DCM, washed with water (2x) and brine. The organic layer was dried and concentrated. The crude product (104-2) was used directly in the next step without further purification. LC-MS, ES-: 550.2 [M-H] - .

[0739] Step 3:

[0740] A mixture of crude (3R,5'S)-1'-((S)-2-(4,6-difluoro-N-methyl-1H-indole-2-carboxamide)-5- methylhexanoyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (104-2) (0.121 g, 0.22 mmol) and Et3N (0.307 mL, 2.20 mmol) in DCM (2.9 mL) was treated with TFAA (0.155 mL, 1.100 mmol) at 0 °C and stirred at room temperature for 30 min. The reaction mixture was diluted with DCM, washed with saturated NaHCO3, water and brine, dried, and concentrated. Purification of the resulting reaction residue on silica gel chromatography with 0-40% acetone / cyclohexane yielded Example 104 (62 mg, 53% yield for 3 steps). LC-MS, ES-: 532.01 [M-H] -1H NMR (400 MHz, Acetone-d6) δ 10.84 (s, 1H), 9.69 (s, 1H), 7.12 - 6.99 (m, 3H), 6.97 - 6.93 (m, 1H), 6.90 (d, J = 7.6 Hz, 1H), 6.86 - 6.78 (m, 1H), 6.74 (td, J = 10.3, 2.1 Hz, 1H), 5.45 (dd, J = 8.8, 6.4 Hz, 1H), 5.22 (t, J = 8.2 Hz, 1H), 4.26 (d, J = 10.7 Hz, 1H), 3.99 (d, J = 10.7 Hz, 1H), 3.46 (s, 3H), 2.74 - 2.64 (m, 2H), 2.04 - 1.91 (m, 2H), 1.74 - 1.63 (m, 2H), 1.62 - 1.48 (m, 2H), 1.47 - 1.36 (m, 2H), 1.36 - 1.25 (m, 2H), 1.25 - 1.15 (m, 2H), 1.15 - 1.05 (m, 2H), 1.05 - 0.95 (m, 2H), 0.95 - 0.85 (m, 2H), 0.85 - 0.75 (m, 2H), 0.75 - 0.65 (m, 2H), 0.65 - 0.55 (m, 2H), 0.55 - 0.45 (m, 2H), 0.45 - 0.35 (m, 2H), 0.35 - 0.25 (m, 2H), 0.25 - 0.15 (m, 2H), 0.15 - 0.05 (m, 2H), 0.05 - 0.05 (m, 2H).

[0741] The following examples were prepared using similar protocols as described above.

[0742]

[0743]

[0744]

[0745]

[0746]

[0747]

[0748]

[0749] Example 413

[0750]

[0751] Step 1.

[0752]

[0753] To a solution of ((benzyloxy)carbonyl)-L-serine (1.25 g, 5.23 mmol) in DMF (20 ml) at -45 °C was added NaHMDS (1 M in THF) (10.97 ml, 10.97 mmol), the resulting reaction mixture was stirred at -45 °C for 20 minutes, allyl bromide (0.543 ml, 6.27 mmol) (shaken in K2CO3) was added, the reaction mixture was then slowly warmed to room temperature and stirred for 18 hours. The reaction mixture was cooled to -20 °C, quenched with AcOH (0.359 ml, 6.27 mmol), diluted with EtOAc / 1 N HCI, and the organic layer was separated, washed with water, brine, dried, filtered and concentrated. The resulting residue was purified on silica gel with CombiFlash eluting with 0-60% acetone / cyclohexane to yield O-allyl-N-((benzyloxy)carbonyl)-L-serine (1.04 g, 3.72 mmol, 71.3% yield). 1 H NMR (400 MHz, Chloroform-d) δ 7.45 - 7.25 (m, 5H), 5.96 - 5.71 (m, 1H), 5.65 (d, J = 8.5 Hz, 1H), 5.26 - 5.12 (m, 2H), 5.10 (d, J = 3.3 Hz, 2H), 4.49 (dt, J = 7.6, 3.4 Hz, 1H), 3.97 (d, J = 5.8 Hz, 2H), 3.90 (dd, J = 9.5, 3.2 Hz, 1H), 3.68 (dd, J = 9.5, 3.6 Hz, 1H).

[0754] Step 2.

[0755]

[0756] To a mixture of O-allyl-N-((benzyloxy)carbonyl)-L-serine (500 mg, 1.790 mmol), paraformaldehyde (323 mg, 10.74 mmol) in acetonitrile (8 ml) was added pTSA (23.84 mg, 0.125 mmol) and the resulting reaction mixture was stirred at 70 °C for 14 hours, the mixture was cooled to room temperature, filtered with celite and the filtrate was collected, then concentrated. The residue was chased with DCM.

[0757] To the residue was added DCM (8 ml) and TFA (2759 μl, 35.8 mmol), triethylsilane (858 μl, 5.37 mmol) and the resulting reaction mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated and chased with DCM. The resulting mixture was diluted with EtOAc, NaOH (IN) solution and then the pH was adjusted to ~4 with HCl (IN). The organic layer was separated and the aqueous layer was extracted with ethyl acetate (2X). The combined organic layers were washed with brine, dried, filtered and concentrated and the resulting residue was purified on silica gel using CombiFlash with 0-5% MeOH / DCM to yield O-allyl-N-((benzyloxy)carbonyl)-N-methyl-L-serine (287 mg, 0.978 mmol, 54.7% yield).

[0758] Step 3.

[0759]

[0760] To a mixture of O-allyl-N-((benzyloxy)carbonyl)-N-methyl-L-serine (70 mg, 0.239 mmol), (3R,5'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-formic acid hydrochloride (80 mg, 0.239 mmol) and HATU (109 mg, 0.286 mmol) in DCM (2 ml) / DMF (0.4 ml) was added 4-methylmorpholine (121 mg, 1.193 mmol). The resulting reaction mixture was stirred at room temperature for 14 hours, the reaction mixture was concentrated and the residue was diluted with ethyl acetate, washed with water, brine, dried, filtered and concentrated. The residue was purified on silica gel using CombiFlash with 0 to 10% MeOH / DCM to yield benzyl ((S)-3-(allyloxy)-l-((3R,5'S)-5'-carbamoyl-2-oxospiro[indoline-3,3'- pyrrolidine]-l'-yl)-l-oxoprop-2-yl)(methyl)carbamate (163 mg). LC-MS, ES + : 507.22 [M+H].

[0761] Step 4.

[0762]

[0763] To (S)-3-(allyloxy)-1-((3R,5'S)-5'-carbamoyl-2-oxospiro[indoline-3,3'-pyrrolidin]-1'- yl)-1-oxoprop-2-yl)(methyl)carbamate (30 mg, 0.06 mmol), Pd-C (6.39 mg, 6.00 μmol) was added MeOH (1.5 ml) and the resulting reaction mixture was stirred under a hydrogen atmosphere for 1.5 h. The resulting mixture was filtered using celite and the filtrate was concentrated.

[0764] To the filtrate was added 4,6-difluoro-1H-indole-2-carboxylic acid (15 mg, 0.078 mmol), HATU (32 mg, 0.084 mmol), DCM (1 ml) and DMF (0.25 ml) and 4-methylmorpholine (24 mg, 0.240 mmol) and the resulting reaction mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated and the residue was purified on silica gel eluting with 0-10% MeOH / DCM to yield (3R,5'S)-1'-(N-(4,6-difluoro-1H-indole-2-carbonyl)-N-methyl-O-propyl-L-serinyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (36 mg). LC-MS, ES + : 576.2 [M+Na].

[0765] Step 5.

[0766]

[0767] To a solution of (3R,5'S)-1'-(N-(4,6-difluoro-1H-indole-2-carbonyl)-N-methyl-O-propyl-L-serinyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (36 mg, 0.065 mmol) in DCM (2 ml) was added TEA (72.5 μl, 0.52 mmol) and TFAA (45.9 μl, 0.325 mmol) and the resulting reaction mixture was stirred at room temperature for 30 min. The reaction mixture was concentrated, diluted with MeOH (1.2 ml) and NH3 (Conc. 0.8 ml) was added and stirred at room temperature for 30 min. The reaction mixture was concentrated. The residue was purified on silica gel using CombiFlash eluting with 0-60% acetone / cyclohexane to yield N-((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-1-oxo-3-propyloxypropan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2-carboxamide (12 mg). LC-MS, ES + : 558.2 [M+Na].

[0768] Example 414

[0769]

[0770] Step 1 : A solution of (3R,5'S)-1 '-((S)-3-cyclopropyl-2-(methylamino)propyl)- 2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (44 mg, 0.123 mmol) and (S)-2-(4-fluorophenyl)-2-hydroxyacetic acid (22.00 mg, 0.129 mmol) in DMF (0.1 mL) and CH2Cl2(0.4 mL) was treated with N-methylmorpholine (50 μL, 0.455 mmol) and HATU (52 mg, 0.137 mmol). The resulting reaction mixture was stirred at room temperature overnight. The resulting mixture was diluted with dichloromethane and quenched with saturated aqueous sodium bicarbonate. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was loaded onto a 4 g silica gel column and eluted with 0% to 100% acetone in cyclohexane to yield (3R,5'S)-1 '-((S)-3-cyclopropyl-2-((S)-2-(4-fluorophenyl)-2-hydroxy-N- methylamido)propyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (16 mg, 0.031 mmol, 25.5% yield) as a white solid. LC-MS, ES - : 507.36 [M-H].

[0771] Step 2: A solution of (3R,5'S)-1 '-((S)-3-cyclopropyl-2-((S)-2-(4-fluorophenyl)-2- hydroxy-N-methylamido)propyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'- carboxamide (14 mg, 0.028 mmol) in CH2Cl2(0.4 mL) was treated with TEA (40 μL, 0.287 mmol) and TFAA (16 μL, 0.113 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h, then quenched with ammonium hydroxide and stirring was continued for 30 min. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was loaded onto a 4 g silica gel column and eluted with 0% to 100% ethyl acetate in cyclohexane to yield (S)-N-((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-yl)-3-cyclopropyl-1-oxopropan-2-yl)-2-(4-fluorophenyl)-2- hydroxy-N-methylacetamide (12 mg, 0.024 mmol, 89% yield) as a white solid. LC-MS, ES - : 489.34 [M-H].1 H NMR (500 MHz, Methanol-d4) δ 7.35 - 7.18 (m, 3H), 7.03 - 6.86 (m, 5H), 5.37 - 5.27 (m, 2H), 5.05 (t, J = 7.9 Hz, 2H), 3.98 (d, J = 10.6 Hz, 1H), 3.79 (d, J = 10.5 Hz, 1H), 2.93 (s, 3H), 2.67 - 2.52 (m, 2H), 1.88 (dt, J = 14.5, 7.5 Hz, 1H), 1.64 (dt, J = 14.0, 7.0 Hz, 1H), 0.73 (ddt, J = 10.3, 7.4, 3.7 Hz, 1H), 0.57 - 0.44 (m, 2H), 0.22 - 0.10 (m, 2H).

[0772] The following examples were prepared using similar methods as described above.

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779] Example 429

[0780]

[0781] Step 1 : To a solution of methyl L-leucine hydrochloride (200 mg, 1.10 mmol) in THF (3.3 mL) was added 4-methoxybenzaldehyde (300 mg, 2.2 mmol), DIPEA (192 μί, 1.1 mmol) and MgS04(225 mg, 1 / 87 mmol). The resulting reaction mixture was stirred at room temperature overnight. The crude product was filtered over celite and evaporated to dryness. The crude product was taken up in methanol (3.3 mL) and sodium borohydride (83 mg, 2.2 mmol) was added. The reaction mixture was quenched with saturated NH4C1, extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on a silica gel chromatography column with 0-70% ethyl acetate in cyclohexane to yield the desired product (249 mg, 85%).

[0782] Step 2: To a solution of the material from Step 1 (249 mg, 0.94 mmol) in THF (3 mL) and methanol (2 mL) was added LiOH (1 mL, 2 M, 2 mmol). Upon completion, the resulting reaction mixture was acidified to pH 3 with 1 M HC1, extracted with ethyl acetate, washed with brine and dried over sodium sulfate to yield the desired product which was used without purification.

[0783] Step 3: To a solution of the material from Step 3 (35 mg, 0.139 mmol) and the spiro intermediate (37 mg, 0.139 mmol) was added HATU (53 mg, 0.139 mmol) and DIPEA (73 μί, 0.418 mmol). The resulting reaction mixture was stirred at room temperature overnight. The reaction mixture was quenched with water, extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on a silica gel chromatography column with 0-80% acetone in cyclohexane to yield the desired product (41 mg, 63%).

[0784] Step 4: To a solution of the material from Step 3 (41 mg, 0.088 mmol) in DCM (1 mL) was added TFAA (37 μί, 0.265 mmol) and Et3N (74 μί, 0.530 mmol) at 0 °C. The crude product was directly loaded on a silica gel column and chromatographed eluting with 0-80% acetone in cyclohexane to yield EP-037611 (30 mg, 63%). LC-MS, ES+ 543.056 [M+H].1H NMR (400 MHz, Acetone-d6 1H NMR (400 MHz, Acetone-d6) δ 9.65 (s, 1H), 7.38 - 7.26 (m, 3H), 7.19 (d, J = 7.4 Hz, 1H), 7.11 - 7.05 (m, 1H), 7.07 - 6.95 (m, 3H), 6.97 (s, 1H), 5.04 (s, 1H), 4.81 (s, 2H), 4.06 (s, 1H), 4.00 (d, J = 10.7 Hz, 1H), 3.88 (d, J = 10.2 Hz, 1H), 3.82 (s, 3H), 2.63 (t, J = 5.9 Hz, 2H), 2.56 (s, 1H), 1.94 (s, 1H), 1.37 (s, 1H), 0.85 (d, J = 7.1 Hz, 4H), 0.75 (d, J = 5.9 Hz, 2H), 0.62 (s, 1H).19F NMR (400 MHz, Acetone-d6) δ 69.4

[0785] The following examples were prepared using similar methods as described above.

[0786]

[0787]

[0788] Example 119

[0789]

[0790] Step 1

[0791] To a solution of compound (23-5) (64 mg, 0.127 mmol) in dry acetone (0.634 mL) was added K2CO3(26.3 mg, 0.190 mmol) and dimethylsulfonate (18.04 μL, 0.190 mmol) at room temperature. The resulting reaction mixture was then heated and refluxed for 2 h. After 2 h, another portion of dimethylsulfonate (6.0 μL, 0.06 mmol) was added and the reaction mixture was heated again for 3 h. The resulting reaction mixture was concentrated to dryness. The residue was diluted with ethyl acetate, washed with water, brine, dried, and concentrated. The resulting residue was purified on silica gel chromatography with 0-50% acetone / cyclohexane to yield compound (119-1) (53 mg, 81% yield). LC-MS, ES+: 519.14 [M+H] + .

[0792] Step 2

[0793] To a solution of compound (119-1) (51 mg, 0.098 mmol) in dry DCM (0.98 mL) was added Dess-Martin Oxidizer (62.6 mg, 0.148 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 3 h. The resulting reaction residue was purified on silica gel chromatography with 0-55% EtOAc / cyclohexane to yield Example 119 (28 mg, 55% yield). LC-MS, ES+: 517.06 [M+H] + . 1H NMR (400 MHz, Acetone-d6) δ 10.52 (s, 1H), 9.52 (d, J = 1.9 Hz, 1H), 7.75 - 7.69 (m, 1H), 7.23 - 7.16 (m, 3H), 7.03 - 6.95 (m, 2H), 6.92 - 6.85 (m, 2H), 6.40 (dd, J = 7.2, 1.2 Hz, 1H), 4.84 (ddd, J = 9.7, 8.3, 4.8 Hz, 1H), 4.54 (ddd, J = 9.2, 6.1, 2.0 Hz, 1H), 4.12 (d, J = 10.4 Hz, 1H), 3.96 (d, J = 10.4 Hz, 1H), 3.79 (s, 3H), 3.07 (s, 3H), 2.37 - 2.29 (m, 1H), 2.20 (dd, J = 13.1, 6.1 Hz, 1H), 1.71 (ddd, J = 14.5, 9.8, 4.2 Hz, 2H), 1.66 - 1.58 (m, 1H), 0.84 (dd, J = 10.7, 6.4 Hz, 6H).

[0794] Example 120

[0795]

[0796] Step 1

[0797] To a solution of Example 119 (24 mg, 0.046 mmol) in dry DMSO (0.186 mL) was added hydroxylamine hydrochloride (4.36 mg, 0.063 mmol). The reaction mixture was stirred at room temperature for 1 h, diluted with ethyl acetate, washed with water (2x), brine, dried, and concentrated to yield the crude oxime intermediate (118-1) (21 mg) which was used directly in the next step. LC-MS, ES+: 532.13 [M+H] + .

[0798] Step 2

[0799] To a solution of the crude oxime intermediate (120-1) (21 mg, 0.046 mmol) in dry acetonitrile (0.79 mL) was added Cu(OAc)2(1.4 mg, 7.9 μmol). The reaction mixture was heated to 70 °C for 1 h and then concentrated. The resulting residue was purified on silica gel chromatography using 0 to 50% acetone / cyclohexane to yield Example 120 (8 mg, 40% yield). LC-MS, ES+: 514.09 [M+H] + . 1H NMR (400 MHz, Acetone-d6) δ 10.60 (s, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.35 (dd, J = 2.3, 0.8 Hz, 1H), 7.29 (td, J = 7.7, 1.2 Hz, 1H), 7.20 - 7.08 (m, 3H), 7.02 (d, J = 7.8 Hz, 1H), 6.95 (td, J = 7.6, 1.0 Hz, 1H), 6.55 (dd, J = 7.4, 1.0 Hz, 1H), 5.17 (t, J = 8.3 Hz, 1H), 4.91 (ddd, J = 9.8, 8.2, 4.6 Hz, 1H), 4.34 (d, J = 10.3 Hz, 1H), 4.05 (d, J = 10.4 Hz, 1H), 3.95 (s, 3H), 3.24 (s, 3H), 2.70 (dd, J = 8.3, 3.9 Hz, 2H), 1.85 (ddd, J = 12.7, 9.4, 4.7 Hz, 2H), 1.73 (dt, J = 9.4, 5.3 Hz, 1H), 0.99 (dd, J = 15.9, 6.4 Hz, 6H).

[0800] Example 121

[0801]

[0802]

[0803] Step 1

[0804] To a mixture of Example 42 (30 mg, 0.058 mmol) in dry acetone (0.29 mL) was added K2CO3(12.11 mg, 0.088 mmol) and dimethylsulfonate (8.31 μL, 0.088 mmol) at room temperature. The resulting reaction mixture was then heated to reflux for 3 hours. The reaction mixture was then concentrated to remove the acetone, the reaction mixture was diluted with ethyl acetate, washed with water, brine, dried and concentrated. The residue was purified on a silica gel column with 0-50% acetone / cyclohexane to yield Example 121 (16 mg, 81% yield). LC-MS, ES-: 526.03 [M-1]. 1H NMR (400 MHz, Acetone-d6) δ 10.36 (s, 1H), 7.20 - 7.10 (m, 2H), 7.10 - 7.03 (m, 2H), 7.00 - 6.91 (m, 2H), 6.87 (t, J = 7.5 Hz, 1H), 6.55 (d, J = 7.7 Hz, 1H), 5.57 (dd, J = 9.6, 5.6 Hz, 1H), 5.20 (t, J = 8.1 Hz, 1H), 4.25 (d, J = 10.7 Hz, 1H), 4.00 (d, J = 10.6 Hz, 1H), 3.97 (s, 3H), 3.45 (s, 3H), 3.22 (s, 3H), 2.77 - 2.63 (m, 2H), 1.93 (ddd, J = 14.4, 9.6, 5.1 Hz, 1H), 1.77 (ddd, J = 14.2, 8.7, 5.6 Hz, 1H), 1.62 (dtd, J = 8.6, 6.6, 5.0 Hz, 1H), 0.98 (dd, J = 23.1, 6.6 Hz, 6H).

[0805] Example 122

[0806]

[0807]

[0808] Step 1

[0809] Compound (1-3) (425 mg, 1.38 mmol) was suspended in DCM (5 mL). Et3N (0.54 mL, 3.9 mmol) and TFAA (0.36 mL, 2.57 mmol) were added dropwise. The reaction mixture was stirred at room temperature for 30 min. A second portion of Et3N (0.2 mL) was added, followed by TFAA (0.12 mL). The reaction mixture was stirred at room temperature for 20 min and concentrated. The residue was purified on a silica gel column to yield compound (122-1) (320 mg, 80%). ESI-MS m / z = 314.05 [M+H] +

[0810] Step 2:

[0811] A mixture of lutidine (0.18 mL, 1.05 mmol) in DCM (1 mL) was cooled to 0 °C. TMSOTf (0.2 mL, 0.95 mmol) was added and the reaction mixture was stirred at 0 °C for 5 min. In another test tube, a mixture of compound (122-1) (100 mg, 0.32 mmol) in DCM (1 mL) was cooled to 0 °C. The TMSOTf / lutidine solution (1.9 mL) was added dropwise and the resulting reaction mixture was stirred at 0 °C for 20 min. NaHC03(4 mL) aqueous solution was added and the reaction mixture was stirred for 10 min and extracted with DCM (2x). The combined organic phase was washed with CsF (0.5 M) aqueous solution and brine, dried over Na2S04and concentrated to give compound (122-2) (68 mg, 100%) as a yellow solid. ESI-MS m / z = 213.88 [M+H] + .

[0812]

[0813] Step 3

[0814] A mixture of leucine tert-butyl ester hydrochloride (1.0 g, 4.47 mmol) and benzyl isocyanate (595 mg, 4.47 mmol) in DCM (6 mL) was prepared. TEA (1.25 mL, 8.95 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 3 h and concentrated. The resulting residue was purified on silica gel to give compound (122-3) (1.5 g) as a colorless paste. ESI-MS m / z = 321.07 [M+H] +.

[0815] Step 4

[0816] To a solution of compound (122-3) (1.5 g) in DCM (12 mL) was added TFA (1.27 mL, 23 mmol). The reaction mixture was stirred at room temperature overnight and concentrated. The resulting residue was purified on silica gel to give compound (122-4) (301 mg, 25% over two steps) as a yellow oil. ESI-MS m / z = 265.02 [M+H] +.

[0817] Step 5

[0818] To a solution of compound (122-2) (20 mg, 0.094 mmol) and compound (122-4) (32 mg, 1.122 mol) in DMF (1 mL) was added TCFH (39 mg, 0.14 mmol) and methyl imidazole (23 mg, 0.38 mmol). The resulting reaction mixture was stirred at room temperature for 15 min, diluted with ethyl acetate, and washed with water and brine. The organic layer was dried over sodium sulfate and concentrated. The residue was purified on silica gel to yield example 122 (30 mg, 70%) as yellow oil. ESI-MS m / z = 460.31 [M+H] + ; 1 H NMR (400 MHz, Chloroform-d) δ 9.06 (br, 1H), 7.21 (d, J = 4.3 Hz, 4H), 7.18 - 7.11 (m, 1H), 7.06 (t, J = 7.8 Hz, 1H), 6.85 (d, J = 7.6 Hz, 1H), 6.83 - 6.73 (m, 1H), 6.65 (d, J = 7.9 Hz, 1H), 6.03 (br, 1H), 5.61 (br, 1H), 4.63 (d, J = 7.8 Hz, 1H), 4.45 (t, J = 8.3 Hz, 1H), 4.33 (d, J = 14.6 Hz, 1H), 4.20 (dd, J = 20.2, 12.6 Hz, 2H), 3.85 (d, J = 10.3 Hz, 1H), 2.72 - 2.58 (m, 1H), 2.24 (dd, J = 13.0, 8.0 Hz, 1H), 1.80 - 1.46 (m, 3H), 0.98 - 0.81 (m, 6H).

[0819] Example 433

[0820]

[0821] Step 1: To a solution of N-((benzyloxy)carbonyl)-N-methyl-L-leucine (300 mg, 1.07 mmol) in MeOH (10 mL) was added Pd / C (w / w 10%, 23 mg, 0.02 eq). After degassing, a hydrogen balloon was introduced. The resulting mixture was stirred at room temperature for 1 h, LCMS showed the reaction was complete. The mixture was filtered and the filtrate was concentrated. The crude product was used directly in the next step.

[0822] Step 2: The crude product from step 1 (N-Me-leucine) and (isocyanomethyl)benzene (0.15 g, 1.12 mmol, 1.05 eq) were mixed together in pyridine (5 mL) and stirred at room temperature for 3 h. It was filtered and concentrated. The crude product was used directly in the next step. ESI-MS m / z = 279.07 [M+H] + .

[0823] Step 3, To the crude product of Step 2 (037625-1) (125 mg, calc. 0.45 mmol) and intermediate 122-2 (48 mg, 0.225 mmol) in DMF (2 mL) was added N-(chloro(dimethylamino)methylene)-N-methylmethanamine hexafluorophosphate (126 mg, 0.45 mmol) and 1 -methyl- 1 H-imidazole (92 mg, 1.13 mmol). The resulting mixture was stirred at room temperature and concentrated. The crude product was purified on a silica gel column to yield the title compound (40 mg, 38%). ESI-MS m / z = 472.19, [M - 1]. 1 HNMR (500 MHz, Acetone-d6) δ 9.71 (s, 1H), 7.27 (td, J = 7.5, 1.7 Hz, 1H), 7.25 - 7.18 (m, 2H), 7.20 - 7.14 (m, 1H), 7.12 - 7.06 (m, 2H), 7.05 - 6.96 (m, 3H), 6.19 (t, J = 5.9 Hz, 1H), 5.31 (dd, J = 9.3, 5.8 Hz, 1H), 5.15 (t, J = 8.4 Hz, 1H), 4.37 (dd, J = 10.7, 1.3 Hz, 1H), 4.22 - 4.10 (m, 2H), 3.91 (d, J = 10.7 Hz, 1H), 2.92 (s, 3H), 2.82 - 2.78 (m, 1H), 2.72 (ddd, J = 13.1, 8.5, 1.2 Hz, 1H), 2.66 (dd, J = 13.2, 8.2 Hz, 1H), 1.73 (ddd, J = 14.3, 9.4, 5.2 Hz, 1H), 1.62 (ddd, J = 14.0, 8.5, 5.9 Hz, 1H), 1.52 (dddd, J = 15.1, 11.8, 7.6, 5.9 Hz, 1H), 1.31 (s, 1H), 0.95 (dd, J = 12.2, 6.6 Hz, 6H).

[0824] Example 43

[0825]

[0826]

[0827] Step 1, To a solution of N-((benzyloxy)carbonyl)-N-methyl-L-leucine (3.0 g, 10.74 mmol) in DCM (30 mL) was added tert-butyl alcohol (2.05 mL, 21.48 mmol) and DCC (2.66 g, 12.89 mmol). The resulting reaction mixture was stirred at room temperature and filtered. The filtrate was concentrated. The crude product was purified on a silica gel column to yield Cbz-N-Me-L-Leu-OtBu (3.2 g, 89%). ESI-MS m / z = 336.03 [M+H] + .

[0828] Step 2: To a solution of Cbz-N-Me-L-Leu-OtBu (3.2 g, 9.54 mmol) in MeOH (30 mL) was added Pd / C (w / w 10%, 355 mg, 0.035 eq). After degassing, the resulting mixture was stirred under hydrogen atmosphere at room temperature for 1 h. LCMS showed the reaction was complete. The solvent was removed to yield the desired product N-Me-L-Leu-OtBu (1.68, 87%). ESI-MS m / z = 202.02 [M+H] + .

[0829] Step 3, Bis(trichloromethyl)carbonate (103 mg, 0.35 mmol) was dissolved in DCM (3 mL). At 0 °C, a solution of N-Me-L-Leu-OtBu (200 mg, 0.99 mmol) and TEA (0.54 mL, 3.97 mmol) in DCM (2 mL) was added and the reaction mixture was stirred at 0 °C for 30 min. A solution of 1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole dihydrochloride (I81 mg, 0.99 mmol) and TEA (0.45 mL) in THF (3 mL) was added. The resulting reaction mixture was stirred at room temperature. After concentration, the crude product was purified on a silica gel column to yield (037826-1) (225 mg, 67%). ESI-MS m / z = 337.12 [M+H] + .

[0830] Example 434 was synthesized following the procedure described in Example 122 starting from (434-1). [M-1], 473.97.

[0831] Example 435

[0832]

[0833] Step 1

[0834] To a stirred solution of compound 1-4 (4.65 g, 17.37 mmol, 1.0 eq) and (S)-2-(((benzyloxy)carbonyl)amino)-4,4-dimethylpentanoic acid (1.1 eq) in CH2Cl2(80 mL) and DMF (8 mL) was added DIEA (3 eq) and HATU (1.1 eq) at room temperature. The resulting reaction mixture was stirred for 1 h at room temperature. The reaction was quenched with 10% citric acid at room temperature. The reaction mixture was extracted with CH2Cl2. The combined organic phase was washed with brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with cyclohexane / acetone (0-50%) to give the desired product as off-white solid. (ES, m / z): [M+H]+= 493.35.

[0835] Step 2

[0836] To a stirred solution of 435-1 (500.00 mg, 1.015 mmol, 1.00 eq) in 10 mL MeOH was added Pd / C (10%, 50 mg) in a 50 mL round bottom flask under nitrogen atmosphere. The reaction mixture was hydrogenated using a hydrogen balloon for 1 h at room temperature under hydrogen atmosphere, filtered over a celite pad and concentrated under reduced pressure to give the desired product. (ES, m / z): [M+H] + = 359.25.

[0837] Step 3

[0838] To a stirred solution of 435-2 (79.74 mg, 0.222 mmol, 1 eq) and DIEA (57.50 mg, 0.444 mmol, 2 eq) in CH2Cl2(2 mL) was added a solution of isocyanobenzene (26.5 mg, 0.222 mmol, 1 eq) in CH2Cl2(3 mL) dropwise at 0 °C. The reaction was monitored by LC-MS until complete conversion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with CH2Cl2and concentrated under vacuum to give the desired product as yellow solid (95.5 mg, 89.89%) which was used in the next step without further purification.

[0839] Step 4

[0840] A mixture of 435-3 (95.5 mg, 0.200 mmol, 1 equiv), DIEA (206.77 mg, 1.600 mmol, 8 equiv) and T3P (763.53 mg, 1.200 mmol, 6 equiv, 50%) in ethyl acetate (1 mL) was stirred at 80 °C for 1 h. The reaction mixture was extracted with ethyl acetate (30 mL) and the organic layer was concentrated under vacuum. The residue was purified using reverse phase flash chromatography to yield the title compound (21.5 mg, 23.40%) as a white solid. [M+H] + = 460.30, 1H NMR (400 MHz, Methanol-d4) δ 1.03 (s, 9H), 1.56 - 1.71 (m, 1H), 1.82 (dd, J = 14.5, 4.3 Hz, 1H), 2.69 (d, J = 8.0 Hz, 2H), 3.98 (d, J = 10.3 Hz, 1H), 4.35 (d, J = 10.3 Hz, 1H), 4.61 (dd, J = 8.5, 4.2 Hz, 1H), 5.17 (t, J = 8.0 Hz, 1H), 6.77 - 7.09 (m, 3H), 7.13 - 7.76 (m, 6H).

[0841] The following examples were prepared using similar methods as described above.

[0842]

[0843]

[0844] Example 44

[0845]

[0846]

[0847] Step 1:

[0848] A round bottom flask was flushed with nitrogen and maintained under a nitrogen atmosphere, to which was added Boc-Asp-Ome 1 (20 g, 81 mmol), DMF (300 ml), cesium carbonate (52.7 g, 162 mmol) and benzyl bromide (11.55 ml, 97 mmol). The resulting solution was stirred at room temperature for 2 h, then diluted with ethyl acetate. The resulting mixture was washed with water 3 times and brine 3 times, then dried over anhydrous sodium sulfate, filtered and concentrated to yield crude product 2. The crude product was carried directly to the next step.

[0849]

[0850] Step 2:

[0851] A round bottom flask was flushed with nitrogen and maintained under a nitrogen atmosphere, to the round bottom flask containing the crude product 2 (27.3 g, 81 mmol) was added DMF (162 ml), silver oxide (56.3 g, 243 mmol) and methyl iodide (101 ml, 1618 mmol). The resulting reaction solution was heated at 60 °C for 1 hour and then diluted with ethyl acetate. The resulting mixture was washed with water 3 times and brine 3 times, then dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (ethyl acetate / cyclohexane 0-20%) to yield 3 (17 g, 60% yield over two steps). 1 H NMR (400 MHz, Chloroform-d) δ 7.44 - 7.27 (m, 5H), 5.27 - 5.10 (m, 2H), 4.86 - 4.56 (m, 1H), 3.73 - 3.65 (m, 3H), 3.13 (dd, J = 16.5, 6.4 Hz, 1H), 3.01 - 2.85 (m, 3H), 2.78 (dq, J = 15.7, 8.0 Hz, 1H), 1.41 (d, J = 12.7 Hz, 9H).

[0852]

[0853] Step 3:

[0854] Palladium on carbon (2.120 g, 10 wt%, 1.992 mmol) was added to a round bottom flask under a nitrogen atmosphere. A solution of 3 (7.0 g, 19.92 mmol) in methanol (100 ml) was added. The reaction flask was evacuated and refilled with hydrogen gas 3 times while stirring at room temperature for 2 hours. The reaction was evacuated and refilled with nitrogen, then filtered over celite and concentrated to yield crude 4 (5.2 g, 100% yield.) The crude product was carried on to the next step. 1 H NMR (400 MHz, Chloroform-d) δ 10.24 (br s, 1H), 4.64 (dt, J = 26.4, 6.9 Hz, 1H), 3.67 (dd, J = 13.9, 6.9 Hz, 3H), 3.07 (dd, J = 15.6, 6.7 Hz, 1H), 2.97 - 2.80 (m, 3H), 2.80 - 2.52 (m, 1H), 1.43 (s, 9H).

[0855]

[0856] Step 4:

[0857] To a round bottom flask containing a solution of compound 4 (967 mg, 3.70 mmol) in THF (35 mL) was added dropwise a solution of methylmagnesium bromide (7.4 mL, 22.2 mmol, 3.0 M in Et20) at -78 °C under a nitrogen atmosphere. The resulting mixture was stirred at the same temperature for 1 h, then allowed to warm to room temperature and stirring was continued for 1 h. The reaction was quenched with saturated ammonium chloride solution and extracted with DCM three times. The organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (MeOH:DCM 0-10%) to yield 6 (85 mg, 9%). 1 H NMR (400 MHz, Chloroform-d) δ 11.10 (s, 1H), 4.65 (q, J = 6.3 Hz, 1H), 3.31 (ddd, J = 17.6, 10.9, 6.5 Hz, 1H), 2.95 (d, J = 6.3 Hz, 3H), 2.80 (dd, J = 17.6, 6.1 Hz, 1H), 2.22 (d, J = 2.3 Hz, 3H), 1.45 (d, J = 7.7 Hz, 9H).

[0858]

[0859] Step 5:

[0860] 5 (85 mg, 0.347 mmol) and (3R,5'S)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (93 mg, 0.347 mmol) were poured into DCM (1.16 mL) and DMF (0.23 mL). At 0 °C, 4-methylmorpholine (114 μΐ, 1.04 mmol) was added and stirred for 5 min. HATU (132 mg, 0.347 mmol) was added at the same temperature. The resulting mixture was stirred at 0 °C for 5 min, then allowed to warm to room temperature and stirred for 2 h. The resulting mixture was diluted with DCM and washed with saturated aqueous sodium bicarbonate, 1 M HC1 and brine. The HC1 aqueous layer was extracted with DCM three times. The collected organic layers were dried over sodium sulfate, filtered and concentrated. The crude product residue was used directly in the next step. Observed M+H = 458.92.

[0861]

[0862] Step 6:

[0863] Compound 6 (159 mg, 0.347 mmol) was treated with HC1 (1.73 mL, 6.94 mmol, 4.0 M in dioxane). The resulting mixture was stirred at room temperature for 30 min and then volatile material was removed under a stream of nitrogen. The crude product salt was used directly in the next step. Observed M+H = 340.87.

[0864]

[0865] Step 7:

[0866] 7 (137 mg, 0.347 mmol) and 4,6,-difluoro-lH-indole-2-carboxylic acid (68 mg, 0.347 mmol) were charged into DCM (1.5 mL) and DMF (0.3 mL). 4-Methylmorpholine (114 μΐ, 1.04 mmol) was added and stirred for 5 min at room temperature. HATU (132 mg, 0.347 mmol) was added. The reaction mixture was stirred for 2 h, diluted with DCM and washed with saturated aqueous sodium bicarbonate, 1 M HC1 and brine. The resulting aqueous layer was extracted with DCM three additional times. The collected organic layers were passed through a phase separator and concentrated. The crude product salt was used directly in the next step. Observed M+H = 537.84.

[0867]

[0868] Step 8:

[0869] Crude 8 (187 mg, 0.347 mmol) was dissolved in water (1.74 mL) / acetonitrile (1.74 mL) under a nitrogen atmosphere. 2,2-Dichloroacetonitrile (0.56 mL, 6.94 mmol) was added followed by palladium (II) trifluoroacetate (115 mg, 0.347 mmol). The resulting mixture was heated to 65 °C and stirred for 15 min. The reaction was diluted with DCM and brine, extracted with DCM 3 times. The collected organic layers were passed through a phase separator and concentrated. The residue was purified by RP HPLC (0.1% TFA / MeCN / 0.1% TFA / water 20-95%) to yield 30 mg of 9, Example 442 (17%). Observed M+Na = 541.7. 1 H NMR (400 MHz, Acetone-d6) δ 10.79 (s, 1H), 9.66 (s, 1H), 7.12 - 6.96 (m, 3H), 6.96 - 6.65 (m, 4H), 5.80 (dd, J = 8.9, 5.2 Hz, 1H), 5.21 (t, J = 8.2 Hz, 1H), 4.20 (s, 1H), 3.97 (d, J = 10.6 Hz, 1H), 3.52 (dd, J = 17.2, 9.0 Hz, 1H), 3.40 (s, 3H), 2.96 - 2.68 (m, 3H), 2.23 (s, 3H).

[0870] Example 443

[0871]

[0872] Example 442 (30 mg, 0.058 mmol) was dissolved in THF (0.58 mL) in a test tube under a nitrogen atmosphere. The solution was cooled to -78 °C and a methylmagnesium bromide solution (58 μΐ, 0.173 mmol, 3.0 M in Et20) was added dropwise. The resulting mixture was stirred at the same temperature for 1 h, then allowed to warm to room temperature and stirring was continued for 1 h. The reaction was quenched with saturated ammonium chloride solution, extracted with DCM three times and dried over sodium sulfate. The organic layer was filtered, concentrated and purified with Shimadzu preparative HPLC (0-95% 0.1% FA in H20 and 0.1% FA in acetonitrile) to yield Example 443 (2 mg, 6%). M+H observed = 535.9. 1 H NMR (400 MHz, Acetone-d6) δ 11.16 (s, 1H), 9.69 (s, 1H), 7.36 - 7.22 (m, 1H), 7.18 - 7.06 (m, 3H), 7.06 - 6.84 (m, 2H), 6.84 - 6.49 (m, 1H), 5.23 (t, J=8.2 Hz, 1H), 4.10 - 3.81 (m, 2H), 3.45 (s, 3H), 2.72 (dd, J=18.5, 8.4 Hz, 2H), 2.56 - 2.38 (m, 1H), 2.25 (s, 1H), 1.95 - 1.81 (m, 1H), 1.21 (s, 6H).

[0873] Example 444

[0874]

[0875] Example 444 was synthesized according to a similar procedure described in Example 443.

[0876] Example 123

[0877]

[0878]

[0879] Step 1

[0880] Compound (1-2) (5.00 g) was dissolved in acetic acid (115 mL). To the resulting reaction solution was added sulfuryl chloride (2.09 g) slowly at room temperature. The reaction mixture was stirred at room temperature overnight, then the reaction mixture was concentrated. The resulting crude product was dissolved in methylene chloride (100 mL) and triethylamine (5.84 g, 8.05 mL, 4.0 equiv) was added, followed by the addition of tert-butyl hydrogencarbonate (4.73 g, 1.5 equiv). The organic layer was then washed with 1 M HC1 (2 x 50 mL), then brine (100 mL), then dried over magnesium sulfate. After concentration, the resulting crude product was purified by RP HPLC to yield compound (123-1) (2.81 g, 51% yield). [M+H] + , 381.1.

[0881] Step 2

[0882] Compound (123-1) (2.81 g) was dissolved in 7 M methanolic amine (36.1 mL) in a 100 mL pressure bottle. The resulting mixture was heated to 60 °C for 36 hours. After concentration, the resulting crude product was triturated with acetonitrile to yield compound (123-2) (1.92 g, 71% yield) as a colorless solid. [M+H] + , 366.1.

[0883] Step 3

[0884] Compound (123-2) (1.61 g) was dissolved in 4 M HC1 in 1,4-dioxane (22.0 mL). The resulting reaction mixture was stirred at room temperature for 2 hours. Concentration yielded compound (123-4) (1.33 g) as a white solid, which was used without further purification. [M+H] + , 266.1.

[0885] Step 4

[0886] Compound (123-3) (103.0 mg), compound (123-3b) (98.0 mg), and HATU (149.0 mg) were combined in a 40 mL flask with a stir bar. DMF (2.27 mL) was added, followed by the addition of DIPEA (179 μί). The resulting reaction mixture was stirred at room temperature overnight. Upon completion, the reaction mixture was diluted with ethyl acetate (50 mL), washed with 1 M HC1 (2 x 20 mL) and brine (20 mL), then dried over magnesium sulfate. After concentration, the resulting crude residue was purified by silica gel chromatography (0 to 10% MeOH in DCM) to yield compound (123-4) (58.1 mg, 34% yield). [M+H] + , 497.2.

[0887] Step 5

[0888] Compound (123-4) (58.1 mg) was dissolved in 4 M HCI / 1,4-dioxane (585 μί). The resulting reaction mixture was stirred for 1.5 h. Concentration of the resulting reaction mixture yielded compound (123-5) (51.0 mg). This compound was used in the next step without further purification. [M+H] + , 397.2.

[0889] Step 6

[0890] Compound (123-5) (51.0 mg), compound (121-5b) (26.7 mg), and HATU (51.5 mg) were combined in a 40 mL flask with a stir bar. DMF (785 μί) was added, followed by DIPEA (62 μί). The resulting reaction mixture was stirred at room temperature for 2.5 h. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with 1 M HCI (3 x 20 mL) and brine (20 mL). The organic layer was dried over magnesium sulfate and concentrated. The resulting crude product was purified using silica gel chromatography (0 to 10% MeOH / DCM) to yield compound (123-6) (26.9 mg, 40% yield). [M+H] + , 576.1.

[0891] Step 7

[0892] Compound (123-6) (26.9 mg) was dissolved in a mixture of MeCN (500 μί) and water (500 μί) in a 20 mL flask. 2,2-Dichloroacetonitrile (56 μί) was then added, followed by palladium (II) trifluoroacetate (1.5 mg). The flask was closed and the reaction mixture was heated to 65 °C for 2 h. Additional 2,2-dichloroacetonitrile (56 μί) and palladium (II) trifluoroacetate (1.5 mg) were added and the reaction mixture was heated at 70 °C for 20 min. It was then cooled to room temperature and the reaction mixture was purified by RP HPLC to yield Example 123 as a white solid (10.0 mg, 38% yield). ESIMS m / z = 558.1 [M+H] + . 1H NMR (400 MHz, Acetone-d6, δ ppm): δ 10.96 (s, 1H), 9.80 (s, 1H), 8.18-8.16 (m, 1H), 7.35-7.34 (m, 1H), 7.15-7.09 (m, 3H), 6.97-6.95 (m, 1H) 6.77-6.72 (m, 1H), 5.23 (app t, J = 8.2, 8.2 Hz, 1H), 5.15-5.09 (m, 1H), 4.46 (d, J = 10.5 Hz, 1H), 4.06 (10.5 Hz), 2.85-2.67 (m, 2H), 2.42-2.18 (m, 2H), 1.47 (d, J = 3.2 Hz, 3H), 1.41 (d, J = 3.2 Hz, 3H). 19F-1H 1.41 (d, J = 3.2 Hz, 3H). 19F-1H 1.41 (d, J = 3.2 Hz, 3H).

[0893] The following examples were prepared using similar methods as described above.

[0894]

[0895]

[0896] Example 44

[0897]

[0898] Step 1

[0899] In a 40 mL screw cap vial with stir bar and pressure relief septum, (S)-1-((3R,5'S)-5'- carbamoyl-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-fluoro-4-methyl-1- oxopentan-2-aminium chloride (1.0 equiv) was combined with 3-(trifluoromethoxy)benzoic acid (26.9 mg, 1.15 equiv) and HATU (49.5 mg, 1.15 equiv). Next, DMF (0.76 mL, 0.15 M) was added followed immediately by DIPEA (60 μί, 3.0 equiv). The resulting reaction mixture was stirred at room temperature until complete consumption of starting material was determined by LCMS. The resulting mixture was diluted with DCM (20 mL) and washed with 1.2 M hydrochloric acid and brine. The organic layer was passed through a phase separator and concentrated to yield crude (3R,5'S)-5-chloro-1'-((S)-4-fluoro-4-methyl-2-(3-(trifluoromethoxy)benzylamino)pentyl)- 2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide. [M+H] + 585.3.

[0900] Step 2

[0901] In a 40 mL screw cap vial equipped with a stir bar and pressure relief septum, the (3R,5'S)-5-chloro-l'-((S)-4-fluoro-4-methyl-2-(3-(trifluoromethoxy)benzylamino)pentyl)-2- oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide produced in Step 1 was dissolved in DCM (1.5 mL, 0.075 M). The resulting solution was cooled in an ice bath and triethylamine (126 μL, 8.0 eq) was added followed by trifluoroacetic anhydride (64 μL, 4.0 eq). The resulting reaction mixture was stirred at room temperature for 1 h. At this time, the resulting mixture was diluted with DCM (10 mL) and 2.0 mL of 30% ammonium hydroxide was added. The mixture was shaken gently, and washed with saturated aqueous sodium bicarbonate (7.5 mL). The organic phase was passed through a phase separator and concentrated. The crude product was purified by RPHPLC (MeCN / water, 0.1% TFA) to give the title compound 1 H NMR (400 MHz, Acetone-d6) δ 9.83 (s, 1H), 8.27 (d, J = 7.7 Hz, 1H), 7.91 (dt, J = 7.8, 1.3 Hz, 1H), 7.82 - 7.80 (m, 1H), 7.64 - 7.60 (m, 1H), 7.54 - 7.51 (m, 1H), 7.25 (dd, J = 8.3, 2.1 Hz, 1H), 7.20 (d, J = 2.1 Hz, 1H), 7.01 (d, J = 8.3 Hz, 1H), 5.24 (t, J = 8.3 Hz, 1H), 5.08 - 5.03 (m, 1H), 4.54 (d, J = 10.5, 1H), 4.06 (d, J = 10.5 Hz, 1H), 2.86 - 2.81 (m, 1H), 2.72 (dd, J = 13.3, 8.2 Hz, 1H), 2.41 - 2.22 (m, 2H), 1.50 (d, J = 6.7 Hz, 3H), 1.44 (d, J = 6.7 Hz, 3H). [M-1], 564.7.

[0902] The following examples were prepared using similar methods as described above.

[0903]

[0904]

[0905]

[0906]

[0907]

[0908] Example 454

[0909]

[0910]

[0911] Step 1

[0912] In a 250 mL flame-dried round bottom flask, (S)-2-((tert-butoxycarbonyl)amino)-4- fluoro-4-methylpentanoic acid (3.00 g, 1.0 equiv) was combined with THF (35 mL, 0.34 M) and methyl iodide-d3 (6.00 mL, 8.0 equiv). The resulting solution was cooled in an ice bath, then sodium cyanide (963.0 mg, 90 wt%, 3.0 equiv) was added. The reaction mixture was allowed to warm to room temperature and stirred for 72 h. Upon completion, the reaction mixture was quenched with HC1 (12 mL, 6 M aq., 6.0 equiv) and further diluted with water (35 mL). The aqueous layer was extracted with ethyl acetate and the combined organic phase was dried over sodium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 50% ethyl acetate / cyclohexane) to yield (S)-2-((tert-butoxycarbonyl)(methyl-d3)amino)-4-fluoro-4-methylpentanoic acid as a white solid (2.74 g, 86%). [M-1], 265.2.

[0913] Step 2

[0914] In a 250 mL round bottom flask equipped with a stir bar, (3R,5'S)-5-chloro-2-oxospiro[indolin-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (2.73 g, 1.0 equiv) was combined with (S)-2-((tert-butoxycarbonyl)amino)-4-fluoro-4-methylpentanoic acid (2.40 g, 1.0 equiv) in DCM / DMF (4: 1, 0.25 M, 28.9 mL DCM, 7.2 mL DMF). The resulting mixture was cooled in an ice bath and N-methylmorpholine (3.2 mL, 3.2 equiv) was added. Next, HATU (3.43 g, 1.0 equiv) was added. After 25 min, complete consumption of starting material was observed by LCMS. The reaction mixture was diluted with DCM (150 mL) and washed with saturated aqueous sodium bicarbonate (35 mL), 1.2 M HC1 (45 mL), and brine. The combined organic layer was dried over magnesium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 10% MeOH / DCM) to yield tert-butyl ((S)-l-((3R,5'S)-5'-carbamoyl-5-chloro-2-oxospiro[indolin-3,3'- pyrrolidin]-l'-yl)-4-fluoro-4-methyl-l-oxopentan-2-yl)(methyl-d3)carbamate as a white solid (4.02 g, 87%). [M+1], 514.2.

[0915] Step 3

[0916] In a 250 mL round bottom flask equipped with a stir bar, ((S)-l-((3R,5'S)-5'- carbamoyl-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-l'-yl)-4-fluoro-4-methyl- 1- oxopentan-2-yl)(methyl-d3)carbamate (4.02 g, 1.0 equiv) was treated with a solution of 4 M HC1 in dioxane (39.1 mL, 20 equiv). The resulting reaction mixture was stirred for 1 h while monitoring for complete conversion by LCMS. Concentration of the reaction yielded (3R,5'S)-5-chloro-l'-((S)-4-fluoro-4-methyl-2-((methyl-d3)amino)pentyl)- 2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride as a white solid which was used without purification. [M+H], 414.2.

[0917] Step 4

[0918] In a 250 mL round bottom flask equipped with a stir bar, the crude (3R,5'S)-5- chloro-l'-((S)-4-fluoro-4-methyl-2-((methyl-d3)amino)pentyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-5'-carboxamide hydrochloride (1.0 equiv) produced in Step 3 was combined with 4,6-difluoro-lH-indole-2-carboxylic acid (1.54 g, 1.0 equiv) in DCM / DMF (5:1, 0.2 M, 32.6 mL DCM, 6.5 mL DMF). The resulting mixture was cooled in an ice bath and N-methylmorpholine (2.58 mL, 3.0 equiv) was added followed by HATU (2.97 g, 1.0 equiv). The reaction mixture was stirred for 14 h at which time complete consumption of starting material was observed by LCMS. The reaction mixture was quenched with 45 mL of saturated aqueous sodium bicarbonate solution and the layers were separated. The aqueous layer was extracted with DCM and the combined organic phases were dried over magnesium sulfate. Purification of the crude residue by silica gel chromatography (0 to 80% acetone / cyclohexane) after concentration yielded (3R,5'S)-5-chloro-l'-((S)-2-(4,6-difluoro-N-(methyl-d3)-lH-indole-2-carboxamide)- 4-fluoropentyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (3.59 g, 77%) as a white solid. [M-H], 591.0.

[0919] Step 5

[0920] In a 250 mL round-bottom flask equipped with a stir bar, (3R,5'S)-5-chloro-1'-((S)-2-(4,6-difluoro-N-(methyl-d3)-1H-indole-2-carboxylic acidamine)-4-fluoro-4-methylpentyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (3.59 g, 1.0 equivalent) was dissolved in DCM (40.4 mL, 0.15 M). The resulting solution was cooled in an ice bath and triethylamine (5.06 mL, 6.0 equivalent) was added, followed by trifluoroacetic anhydride (2.57 mL, 3.0 equivalent). The reaction mixture was warmed to room temperature and stirred for 45 min, at which point complete consumption of the starting material was observed by LCMS. The resulting mixture was diluted with 150 mL of DCM and washed (60 mL) with saturated sodium bicarbonate aqueous solution. The aqueous phase was extracted with methylene chloride. The bound organic layer was washed with brine and dried over magnesium sulfate. After concentration, the crude product residue was purified by C18 column (gradient elution, water / MeCN) to produce a white solid N-((S)-1-((3R,5'S)-5-chloro-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidine]-1'-yl)-4-fluoro-4-methyl-1-oxopent-2-yl)-4,6-difluoro-N-(methyl-d3)-1H-indole-2-carboxamide (3.08 g, 88% yield). 1 H NMR (400MHz, acetone-d6) δ10.68 (s, 1H), 9.80 (s, 1H), 7.09-7.06 (m, 1H), 6.97-6. 88 (m, 4H), 6.75 (td, J=10.3, 2.1Hz, 1H), 5.84 (dd, J=7.6, 5.6Hz, 1H), 5.33 (d d, J=8.8, 7.7Hz, 1H), 4.50 (d, J=10.9Hz, 1H), 3.95 (d, J=11.0Hz, 1H), 2.84-2 .65 (m, 2H), 2.52-2.25 (m, 2H), 1.45 (d, J=6.8Hz, 3H), 1.40 (d, J=7.0Hz, 3H). [M-1], 573.2.

[0921] The following examples were prepared using a method similar to that described above.

[0922]

[0923]

[0924]

[0925]

[0926]

[0927]

[0928]

[0929] Example 472

[0930]

[0931]

[0932] Step 1

[0933] To a solution of (S)-methyl 2-(bis(tert-butoxycarbonyl)amino)-5-oxopentanoate (1.10 g, 1.0 equiv), DABCO (1.07 g, 3.0 equiv), and 2-iodo-5-(trifluoromethyl)aniline (1.01 g, 1.1 equiv) in DMF (16 mL, 0.2 M) in a 100 mL Schlenk tube was flushed with nitrogen for 20 minutes. Palladium (II) acetate (72.0 mg, 0.10 equiv) was then added and the reaction mixture was heated to 90 °C for 30 hours under a nitrogen atmosphere. Upon completion, the resulting mixture was diluted with water and the aqueous phase was extracted with ethyl acetate. The combined organic phase was dried over magnesium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 40% ethyl acetate / cyclohexane) to yield (S)-methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(6-(trifluoromethyl)-1H-indol-3-yl)propanoate (1.09 g, 70%). [M+1], 487.2.

[0934] Step 2

[0935] In a 40 mL screw-cap vial equipped with a pressure relief septum and stirring bar, (S)-methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(6-(trifluoromethyl)-1H-indol-3-yl)propanoate (1.09 g, 1.0 equiv) was treated with a 4 M solution of HCl in dioxane (11.2 mL, 20 equiv of HCl) and the resulting reaction mixture was stirred at room temperature for 12 hours. Upon completion, the reaction mixture was concentrated to yield (S)-methyl 2-amino-3-(6-(trifluoromethyl)-1H-indol-3-yl)propanoate hydrochloride which was used directly in the next step without purification. [M+1], 287.1.

[0936] Step 3

[0937] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, (S)-2-amino-3-(6-(trifluoromethyl)-lH-indol-3-yl)propanoic acid methyl ester hydrochloride (722 mg, 1.0 equiv) was combined with formaldehyde (183 μL, 37 wt%, 1.1 equiv) in MeOH (4.5 mL, 0.5 M). The reaction mixture was heated at 65 °C for 3 h. After the reaction was judged complete by LCMS, the reaction mixture was concentrated to yield crude (S)-7-(trifluoromethyl)-2,3,4,9-tetrahydro-lH-pyrrolo[3,4- b]indole-3-carboxylic acid methyl ester hydrochloride which was used directly in the next step without purification. [M+1], 299.1.

[0938] Step 4

[0939] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, (S)-7-(trifluoromethyl)-2,3,4,9-tetrahydro-lH-pyrrolo[3,4-b]indole-3-carboxylic acid ester hydrochloride (1.0 equiv) was suspended in DCM (5.6 mL, 0.4 M). Triethylamine (2.0 equiv, 623 μL) was added followed by Boc-anhydride (1.23 mL, 1.1 equiv, 2 M in DCM). The resulting reaction mixture was stirred at room temperature for 20 h, then the reaction mixture was concentrated and purified with silica gel chromatography to yield 2-(tert-butyl) 3-methyl (S)-7-(trifluoromethyl)-l,3,4,9-tetrahydro-2H-pyrrolo[3,4- b]indole-2,3-dicarboxylate (681 mg, 76%). [M-1], 397.0.

[0940] Step 5

[0941] In a 40 mL screw-cap vial equipped with a pressure relief septum and stir bar, 2-(tert-butyl) 3-methyl (S)-7-(trifluoromethyl)-l,3,4,9-tetrahydro-2H-pyrrolo[3,4-b]indole-2,3-carbonate (681.0 mg, 1.0 equiv) was suspended in a mixture of THF, water, and acetic acid (10 mL THF, 1 mL DI water, 685 μL acetic acid) at -40 °C, to this solution was added N-bromosuccinimide (304 mg, 1.0 equiv) in portions. After 2.5 h, the mixture was warmed to 0 °C in an ice bath and an additional portion of acetic acid (685 μL) was added. After 20 min, complete consumption of starting material was observed by LCMS and 75 mL of saturated aqueous sodium bicarbonate solution was added slowly to the reaction mixture. The reaction mixture was extracted with ethyl acetate and the combined organic phase was dried over magnesium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 100% MTBE / cyclohexane) to yield 1'-(tert-butyl) 5'-methyl (3R,5'S)-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidine]- 1',5'-carbonate (591.7 mg, 84%, 6:1 diastereomeric mixture) as a colorless oil. [M-1], 413.0.

[0942] Step 6

[0943] In a 25 mL pressure tube equipped with a stir bar, the 1'-(tert-butyl) 5'-methyl (3R,5'S)-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidine]-l',5'-carbonate (591.7 mg, 1.0 equiv) produced in Step 5 was dissolved in 7 M NH3in MeOH (6.1 mL, 30 equiv NH3). The resulting reaction mixture was heated to 60 °C for 48 h. At this time, the reaction mixture was concentrated and the resulting crude product was purified by RP HPLC (MeCN / water, 0.1% TFA) to yield the single diastereomer (3R,5'S)-5'-carbamoyl-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidine]-l'-carboxylic acid tert-butyl ester (311.0 mg, 55%). [M+Na], 422.1.

[0944] Step 7

[0945] In a 40 mL screw cap vial, equipped with a pressure relief septum and stir bar, treat (3R,5'S)-5'-carbamoyl-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-1'- carboxylic acid tert-butyl ester (311.0 mg, 1.0 eq) with 4M HCI in dioxane (3.9 mL, 20 eq HCI). Stir the resulting reaction mixture at room temperature for 1 h and concentrate to yield (3R,5'S)-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride as a white solid (261.0 mg). [M+1], 300.1.

[0946] Step 8

[0947] In a 40 mL screw cap vial, equipped with a pressure relief septum and stir bar, treat (3R,5'S)-5'-carbamoyl-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-1'- carboxylic acid tert-butyl ester (311.0 mg, 1.0 eq) with 4M HCI in dioxane (3.9 mL, 20 eq HCI). Stir the resulting reaction mixture at room temperature for 1 h and concentrate to yield (3R,5'S)-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride as a white solid (261.0 mg). [M+1], 300.1.

[0948] Step 9

[0949] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, treat ((S)-1-((3R,5'S)-5'-carbamoyl-2-oxo-6-(trifluoromethyl)spiro[indolin-3,3'- pyrrolidin]-1'-yl)-4-fluoro-4-methyl-1-oxopentan-2-yl)(methyl-d3)carbamate (107 mg, 1.0 equiv) with a solution of 4 M HCI in dioxane (973 μL, 20 equiv HCI) at room temperature. After 1 h, concentrate the reaction mixture to yield (3R,5'S)-1'-((S)-4-fluoro-4-methyl-2-((methyl-d3)amino)pentyl)-2-oxo-6- (trifluoromethyl)spiro[indolin-3,3'-pyrrolidin]-5'-carboxamide hydrochloride which is used in the next step without further purification. [M+1], 448.2.

[0950] Step 10

[0951] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, combine (3R,5'S)-1'-((S)-4-fluoro-4-methyl-2-((methyl-d3)amino)pentyl)-2-oxo-6- (trifluoromethyl)spiro[indolin-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (1.0 equiv) produced in Step 9 with 4,6-difluoro-1H-indole-2-carboxylic acid (44.1 mg, 1.15 equiv) and HATU (85.0 mg, 1.15 equiv) in DMF (1.3 mL, 0.15 M). Subsequently add DIPEA (102 μL, 3.0 equiv) and stir the resulting reaction mixture at room temperature for 14 h. Filter the crude reaction mixture and purify with RP HPLC (MeCN, water, 0.1% TFA) to yield (3R,5'S)-1'-((S)-2-(4,6-difluoro-N-(methyl-d3)-1H-indole-2-carboxamide)-4-fluoro-4- methylpentyl)-2-oxo-6-(trifluoromethyl)spiro[indolin-3,3'-pyrrolidin]-5'-carboxamide as a white solid (88.3 mg, 72%). [M-1], 625.1.

[0952] Step 11

[0953] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, (3R,5'S)-1 '-((S)-2-(4,6-difluoro-N-(methyl-d3)-1 H-indole-2-carboxamide)-4- fluoro-4-methylpentyl)-2-oxo-6-(trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-5'- carboxamide (88.3 mg, 1.0 equiv) was dissolved in DCM (2.8 mL, 0.05 M) at 0 °C. To the solution was added triethylamine (1.27 mL, 9.0 equiv) followed by trifluoroacetic anhydride (634 μί, 4.5 equiv). The resulting reaction mixture was stirred at room temperature for 1.5 h, after which ammonium hydroxide (250 μί, 30 wt%) was added and the resulting mixture was shaken gently. The reaction mixture was then diluted with saturated aqueous sodium bicarbonate (5 mL) and DCM (5.0 mL) and passed through a phase separator. After concentration, the crude product was purified by RP HPLC (MeCN, water, 0.1% TFA) to give N-((S)-1-((3R,5'S)-5'-cyano-2-oxo-6- (trifluoromethyl)spiro[indoline-3,3'-pyrrolidin]-1 '-yl)-4-fluoro-4-methyl-1 -oxopentan-2-yl)- 4,6-difluoro-N-(methyl-d3)-1 H-indole-2-carboxamide (1.7 mg, 2%). 1 H NMR (400 MHz, Acetone-d6) δ 10.76 (s, 1H), 9.96 (s, 1H), 7.19 - 6.99 (m, 5H), 6.74 (td, J = 10.3, 2.1 Hz, 1H), 5.83 (dd, J = 7.6, 5.5 Hz, 1H), 5.28 (t, J = 8.2 Hz, 1H), 4.52 (d, J = 11.0 Hz, 1H), 4.01 (d, J = 10.9 Hz, 1H), 2.88 - 2.71 (m, 2H), 2.48 - 2.29 (m, 2H), 1.45 (d, J = 6.3 Hz, 3H), 1.40 (d, J = 6.5 Hz, 3H). [M-1], 607.4.

[0954] Example 47

[0955]

[0956] Step 1

[0957] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, treat ((S)-1-((3R,5'S)-5'-carbamoyl-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4- fluoro-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (318.0 mg, 1.0 eq) with 4M HCI in dioxane (3.1 mL, 20 eq HCI). Stir the resulting reaction mixture at room temperature for 1.5 h. Determine by LCMS that the starting material is consumed and concentrate the reaction mixture to yield (3R,5'S)-5-chloro-1'-((S)-4-fluoro-4-methyl-2- (methylamino)pentyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (278 mg) as a white solid which is used in Step 2 without further purification. [M+1], 411.2.

[0958] Step 2 At 20 °C in a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, add a mixture of (3R,5'S)-5-chloro-1'-((S)-4-fluoro-4-methyl-2-(methylamino)pentyl)-2- oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (50.0 mg, 1.0 eq) and 1- (pyridin-4-yl)cyclopropane-1-carboxylic acid (18.2 mg, 1.0 eq) and N-methylmorpholine (40 μL, 3.2 eq) in DCM / DMF (5:1 DCM / DMF, 0.15 M, 620 μL DCM / 120 μL DMF). Subsequently, add HATU (42.5 mg, 1.0 eq) and stir the reaction mixture for 14 h. After completion, add 100 uL formic acid and concentrate the reaction mixture. Purify the crude residue with RP HPLC (MeCN / water / 0.1% TFA) to yield (3R,5'S)-5-chloro-1'-((S)-4-fluoro-4-methyl-2-(N-methyl-1-(pyridin-4-yl)cyclopropyl-1- carboxylic acid amide)pentyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (36.2 mg, 58%). [M+1], 556.4.

[0959] Step 3

[0960] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, (3R,5'S)-5-chloro-l'-((S)-4-fluoro-4-methyl-2-(N-methyl-l-(pyridin-4-yl)cyclopropyl-l- carboxamide)pentyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide (36.2 mg, 1.0 equiv) was dissolved in DCM (870 μL, 0.075 M) and boroguanidine (46.5 mg, 3.0 equiv) was added. The resulting mixture was stirred at room temperature for 14 h. The resulting mixture was diluted with saturated aqueous bicarbonate solution (3 mL) and DCM (5 mL) and passed through a phase separator. After concentration, the residue was purified by RHPLC to yield N-((S)-l-((3R,5'S)-5-chloro-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]- 1'-yl)-4-fluoro-4-methyl-l-oxopentan-2-yl)-N-methyl-l-(pyridin-4-yl)cyclopropyl-l- carboxamide (11.1 mg, 32%) as a white solid. 1 H NMR (400 MHz, Acetone-d6) δ 8.70 (d, J = 6.7 Hz, 2H), 7.54 (d, J = 6.7 Hz, 2H), 7.35 (dd, J = 8.3, 2.1 Hz, 1H), 7.12 (d, J = 2.1 Hz, 1H), 7.07 (d, J = 8.3 Hz, 1H), 5.61 (dd, J = 7.4, 5.6 Hz, 1H), 5.26 (t, J = 8.4 Hz, 1H), 4.24 (d, J = 10.6 Hz, 1H), 4.08 (d, J = 10.6 Hz, 1H), 3.00 (s, 2H), 2.85 - 2.67 (m, 1H), 2.44 - 2.19 (m, 2H), 1.68 - 1.58 (m, 2H), 1.46 (d, J = 7.8 Hz, 3H), 1.41 (d, J = 7.7 Hz, 3H), 1.39 - 1.24 (m, 2H). [M+l], 538.2.

[0961] The following examples were prepared using similar methods as described above.

[0962]

[0963]

[0964]

[0965]

[0966]

[0967]

[0968]

[0969]

[0970] Example 49

[0971]

[0972]

[0973] Step 1:

[0974] (S)-3-cyclopropyl-2-((((2,6-difluorobenzyl)oxy)carbonyl)amino)propanoic acid (431.0 mg, 1.0 eq) was dissolved in DMF (4.3 mL, 0.5 M) in a 40 mL vial equipped with a pressure relief cap and stir bar. The solution was then cooled in an ice bath and CDI (431.0 mg, 1.0 eq) was added. The resulting mixture was allowed to warm to room temperature. After stirring for 20 minutes, (S)-2-amino-3-methylbutanoic acid (100.0 mg, 1.0 eq) was added and the reaction mixture was heated to 55 °C for 14 h. The reaction mixture was then cooled to room temperature, diluted with water and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried over magnesium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 40% ethyl acetate / cyclohexane) to yield (S)-3-cyclopropyl-2-((((2,6-difluorobenzyl)oxy)carbonyl)amino)-3-methylbutanoic acid (123.0 mg, 73%). [M+1], 328.2.

[0975] Step 2

[0976] (S)-3-cyclopropyl-2-((((2,6-difluorobenzyl)oxy)carbonyl)amino)-3-methylbutanoic acid (123.0 mg, 1.0 eq) was dissolved in a mixture of MeOH and water (1 : 1, 0.26 M, 7.6 mL MeOH, 7.6 mL water) in a 50 mL round bottom flask equipped with a stir bar at 0 °C. LiOH (235.0 mg, 2.5 eq) was then added. The reaction mixture was slowly allowed to warm to room temperature and stirred for 14 h. The reaction mixture was concentrated to remove the MeOH and acidified with 6 M HC1. The aqueous layer was extracted with DCM and the combined organic phases were dried over magnesium sulfate. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 70% ethyl acetate / cyclohexane) to yield (S)-3-cyclopropyl-2-((((2,6-difluorobenzyl)oxy)carbonyl)amino)-3-methylbutanoic acid (123.0 mg, 73%). [M+1], 328.2.

[0977] Step 3

[0978] In a 40 mL vial with pressure relief cap and stir bar, (3R,5'S)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide hydrochloride (100.0 mg, 1.0 equiv) and (S)-3-cyclopropyl-2-((((2,6-difluorobenzyl)oxy)carbonyl)amino)propanoic acid (114.0 mg, 1.15 equiv) were combined in DMF (2.2 mL, 0.15 M). Subsequently, HATU (145.0 mg, 1.15 equiv) was added, followed immediately by DIPEA (173 μL, 3.0 equiv). The resulting reaction mixture was stirred at room temperature for 14 h. Upon completion, the reaction was diluted with DCM and washed with 1.2 M HC1 and brine, and passed through a phase separator. Concentration yielded crude 2,6-difluorobenzyl ((S)-1-((3R,5'S)-5'-carbamoyl-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidine]-1'-yl)-3-cyclopropyl-1-oxopropan-2-yl)carbamate which was used in the next step without further purification. [M+1], 547.2.

[0979] Step 4:

[0980] In a 40 mL vial with pressure relief cap and stir bar, crude 2,6-difluorobenzyl ((S)-1-((3R,5'S)-5'-carbamoyl-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidine]-1'-yl)-3-cyclopropyl-1-oxopropan-2-yl)carbamate from Step 3 was dissolved in DCM (4.7 mL, 0.075 M) at 0 °C. To the resulting solution was added triethylamine (346 μL, 7.0 equiv) followed by trifluoroacetic anhydride (200 μL, 4.0 equiv). The reaction was stirred at room temperature for 1 h and quenched with saturated aqueous sodium bicarbonate (5 mL) and 2.0 mL ammonium hydroxide (30 wt %). The reaction was further diluted with DCM (5 mL) and passed through a phase separator. After concentration, the crude residue was purified by RP HPLC (MeCN / water / 0.1% TFA) to yield 2,6-difluorobenzyl ((S)-1-((3R,5'S)-5-chloro-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidine]-1'-yl)-3-cyclopropyl-1-oxopropan-2-yl)carbamate (7.9 mg, 4.2%). 1H NMR (400 MHz, Acetone-d6) δ 9.83 (s, 1H), 7.54 - 7.46 (m, 1H), 7.33 - 7.27 (m, 2H), 7.10 - 7.01 (m, 3H), 6.73 (d, J = 7.6 Hz, 1H), 7.10 - 7.02 (m, 3H), 6.73 (d, J = 7.6 Hz, 1H), 5.26 (t, J = 8.1 Hz, 1H), 5.20 (d, J = 11.8 Hz, 1H), 5.11 (d, J = 11.8 Hz, 1H), 4.51 (q, J = 7.3 Hz, 1H), 4.38 (d, J = 10.5 Hz, 1H), 4.07 (d, J = 10.5 Hz, 1H), 2.88 - 2.81 (m, 1H), 2.71 (dd, J = 13.3, 7.8 Hz, 1H), 1.76 - 1.65 (m, 2H), 0.92 - 0.82 (m, 1H), 0.54 - 0.46 (m, 2H), 0.27 - 0.14 (m, 2H). [M - 1], 527.0.

[0981] The following examples were prepared using similar methods as described above.

[0982]

[0983]

[0984] Example 49

[0985]

[0986]

[0987] Step 1

[0988] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, 1'-(tert- butyl) 5'-methyl (3R,5'S)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidine]-1',5'-carbonate (209.0 mg, 1.0 eq) was treated with 4 M HC1 in dioxane (2.7 mL, 20 eq HC1). The resulting mixture was stirred at room temperature for 2 h and concentrated to yield methyl (3R,5'S)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxylate hydrochloride (174.0 mg), which was used in the next step without further purification. [M + 1], 281.1.

[0989] Step 2

[0990] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, a mixture of N-(tert-butoxycarbonyl)-N-methyl-L-leucine (135.0 mg, 1.0 equiv) in DCM and DMF (5:1 DCM / DMF, 0.25 M, 1.8 mL DCM, 370 μL DMF) was combined with (3R,5'S)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxylic acid methyl ester hydrochloride (174.0 mg, 1.0 equiv). The resulting mixture was cooled in an ice bath and N-methylmorpholine (181 μL, 3.0 equiv) was added followed by HATU (209.0 mg, 1.0 equiv). The resulting reaction mixture was allowed to slowly warm to room temperature and stirred for 14 h. The resulting reaction mixture was diluted with brine (5 mL) and DCM (5 mL) and the organic phase was passed through a phase separator. After concentration, the crude residue was purified by silica gel chromatography (gradient elution, 0 to 55% ethyl acetate / cyclohexane) to yield (3R,5'S)-l'-(N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxylic acid methyl ester (246.6 mg, 88%). [M+1], 508.4.

[0991] Step 3

[0992] In a 40 mL screw cap vial equipped with a pressure relief septum and stir bar, (3R,5'S)-l'-(N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxylic acid methyl ester (246.6 mg, 1.0 equiv) was treated with 4 M HC1 in dioxane (2.43 mL, 20 equiv HC1). The resulting mixture was stirred at room temperature for 2 h and concentrated to yield (3R,5'S)-5-chloro-l'-(methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxylic acid methyl ester hydrochloride as a white solid which was used without purification in the next step. [M+1], 408.2.

[0993] Step 4

[0994] In a 40 mL screw cap vial equipped with a pressure relief bottle and stir bar, the product from Step 3, (3R,5'S)-5-chloro-l'-(methyl-L-leucyl)-2-oxospiro[indoline-3,3'- pyrrolidine]-5'-carboxylic acid methyl ester hydrochloride (1.0 equiv) was combined with 4,6-difluoro-lH-indole-2-carboxylic acid (96.0 mg, 1.0 equiv) in DCM and DMF (5: 1 DCM / DMF, 0.2 M, 2.02 mL DCM, 410 μL DMF). The resulting mixture was cooled in an ice bath and N-methylmorpholine (160 μL, 3.0 equiv) was added followed by HATU (185.0 mg, 1.0 equiv). The reaction was allowed to slowly warm to room temperature and stirred for 14 h. Upon completion, the resulting mixture was diluted with DCM (5 mL) and brine (5 mL) and passed through a phase separator. Upon concentration, the crude residue was purified by silica gel chromatography to yield (3R,5'S)-5-chloro-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxylic acid methyl ester (224.4 mg, 79%). [M-l], 585.1.

[0995] Step 5

[0996] In a 40 mL screw cap vial equipped with a pressure relief bottle and stir bar, (3R,5'S)-5-chloro-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'- pyrrolidine]-5'-carboxylic acid methyl ester (224.4 mg, 1.0 equiv) was dissolved in 1,2-DCE (3.82 mL, 0.1 M). Trimethyltin hydroxide (207.0 mg, 3.0 equiv) was then added and the resulting reaction mixture was heated to 75 °C for 16 h. It was then cooled to room temperature, the reaction mixture was diluted with DCM (20 mL), washed twice with 1.2 M HC1 (5 mL), washed once with brine and the organic layer was passed through a phase separator. Upon concentration, the crude residue was purified by RP HPLC to yield (3R,5'S)-5-chloro-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxylic acid as a white solid (1.44 mg, 0.7%). 1H NMR (400 MHz, Acetone-d6) δ 10.71 (s, 1H), 9.78 (s, 1H), 7.12-7.05 (m, 1H), 7.02-6.95 (m, 3H), 6.89 (d, J = 8.3 Hz, 1H), 6.74 (td, J = 10.3, 2.1 Hz, 1H), 5.59 (t, J = 7.5 Hz, 1H), 4.94 (t, J = 8.8 Hz, 1H), 4.38 (d, J = 10.4 Hz, 1H), 3.97 (d, J = 10.5 Hz, 1H), 3.47 (s, 3H), 2.63-2.46 (m, 2H), 1.81 (t, J = 7.2 Hz, 2H), 1.70-1.58 (m, 1H), 1.00 (d, J = 6.6 Hz, 3H), 0.95 (d, J = 6.6 Hz, 3H). [M-1], 570.9.

[0997] Example 498

[0998]

[0999]

[1000] Step 1 : A clear colorless solution of 1 '-(tert-butyl) 5'-methyl (3R,5'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-1 ',5'-carbonate (3.94 g, 11.4 mmol, dr 10 / 1) in acetonitrile (40 mL) was treated with NBS (2.23 g, 12.5 mmol) in three portions at room temperature. The reaction mixture was stirred at room temperature for 3 h. It turned into a bright yellow solution. LCMS showed no starting material. The reaction mixture was quenched with aqueous Na2S2O3solution. The resulting reaction mixture was continued to stir at room temperature for 30 min. The resulting turbid reaction mixture was further diluted with EtOAc (80 mL). The aqueous layer was extracted with ethyl acetate twice. The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated to give the crude product as an off-white solid. The crude product was dissolved in DCM (10 mL) and filtered through a 80 g silica gel pad (MTBE) to give the desired product as a white solid (dr 10 / 1). The product was treated with MTBE / hexanes (2:1) (30 mL). The mixture was sonicated for 79 min until a milky suspension was generated, the suspension was filtered and washed with MTBE / hexanes (2:1) to give 1 '-(tert-butyl) 5'-methyl (3R,5'S)-5-bromo-2-oxospiro[indoline-3,3'-pyrrolidine]-1 ',5'-dicarboxylate as a white solid (4.23 g, 10.0 mmol, dr >100 / 1, 87% yield). LC-MS, ES - : 422.74, 424.64 [M-H]- .

[1001] Step 2: Treat l'-(tert-butyl) 5'-methyl (3R,5'S)-5-bromo-2-oxospiro[indoline-3,3'- pyrrolidin]-l',5'-carbonate (5.3 g, 12.46 mmol) with 7 N ammonia in methane (40 ml, 280 mmol). Heat the resulting mixture to 50 °C and stir over the weekend. Concentrate the reaction mixture in vacuo to yield (3R,5'S)-5-bromo-5'-carbamoyl-2-oxospiro[indoline-3,3'- pyrrolidin]-l'-carboxylic acid tert-butyl ester as an off-white solid (5.11 g, 12.5 mmol, 100% yield). LC-MS, ES - : 408.10, 410.07 [M-H] - .

[1002] Step 3: To a solution of (3R,5'S)-5-bromo-5'-carbamoyl-2-oxospiro[indoline-3,3'- pyrrolidin]-l'-carboxylic acid tert-butyl ester (2.39 g, 5.83 mmol) in DMF (4.8 ml) at 0 °C, add 4 M HC1 in dioxane (20 ml, 80 mmol) dropwise. Allow the reaction to warm to room temperature and stir for 3 hours. Add the resulting solution to DCM (100 mL) to precipitate the product. Filter the suspension and then dry the solid under vacuum to yield (3R,5'S)-5-bromo-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride as an off-white solid (1.895 g, 5.47 mmol, 94% yield). LC-MS, ES + : 265.14, 267.16 [M+H] + .

[1003] Step 4: A solution of (3R,5'S)-5-bromo-2-oxospiro[indoline-3,3'-pyrrolidin]-5'- carboxamide hydrochloride (2.06 g, 5.94 mmol) and N-(tert-butoxycarbonyl)-N- methyl-L-leucine (1.531 g, 6.24 mmol) in THF (20.00 mL) and DMF (2.0 mL) was treated with N-methylmorpholine (1.960 mL, 17.83 mmol) and 50% T3P in DMF (3.82 mL, 6.54 mmol) at 0 °C. The reaction was allowed to warm to room temperature and stirred for 1 h, then quenched with saturated solution of sodium bicarbonate. The reaction mixture was extracted with ethyl acetate 2 times. The combined organic phase was washed with 1 M HC1, water and brine, dried over sodium sulfate, filtered and concentrated under vacuum to yield ((S)-1-((3R,5'S)-5-bromo-5'-carbamoyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamic acid tert-butyl ester (2.867 g, 5.33 mmol, 90% yield) as off-white solid. LC-MS, ES - : 535.23, 537.27 [M-H] - .

[1004] Step 5: ((S)-1-((3R,5'S)-5-bromo-5'-carbamoyl-2-oxospiro[indoline-3,3'-pyrrolidin]-1'- yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamic acid tert-butyl ester (2.867 g, 5.33 mmol) was treated with 4 M HC1 in dioxane (13.34 mL, 53.3 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was evaporated and dried under high vacuum to yield (3R,5'S)-5-bromo-1'-(methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-5'- carboxamide hydrochloride (2.44 g, 5.15 mmol, 97% yield) as white solid. LC-MS, ES + : 437.31, 439.27 [M+H] + .

[1005] Step 6: A solution of (3R,5'S)-5-bromo-l'-(methyl-L-leucyl)-2-oxospiro[indoline- 3,3'-pyrrolidine]-5'-carboxamide hydrochloride (2.44 g, 5.15 mmol) and 4,6- difluoro-lH-indole-2-carboxylic acid (1.117 g, 5.66 mmol) in DMF (25.7 mL) was treated with HATU (2.350 g, 6.18 mmol) and DIPEA (2.70 mL, 15.45 mmol) at room temperature. The resulting reaction mixture was stirred at room temperature overnight. The resulting reaction mixture was diluted with ethyl acetate and washed with water and saturated sodium chloride solution. The organic layer was dried over sodium sulfate, filtered and concentrated under vacuum. The crude product was loaded onto a silica gel column (40 g) and eluted with 0% to 75% acetone in hexanes to yield (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide as an off-white solid (2.7 g, 4.38 mmol, 85% yield). LC-MS, ES - : 614.39, 616.31 [M-H] - .

[1006] Step 7: A solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (41 mg, 0.067 mmol) in n-PrOH (0.5 mL) was treated with trifluoro(prop-l- ane-2-yl)-l4-borane, potassium salt (16 mg, 0.108 mmol), TEA (30 μL, 0.215 mmol) and PdCl2(dppf) (6 mg, 8.20 μmol) under nitrogen. The resulting mixture was bubbled with nitrogen for 5 minutes. The reaction mixture was warmed to 90 °C and stirred overnight. The resulting mixture was filtered over celite and concentrated under vacuum. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% acetone in cyclohexane to yield (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxo-5-(prop-l-ane-2-yl)spiro[indoline-3,3'-pyrrolidine]-5'-carboxamide as an orange solid (19 mg, 0.033 mmol, 49.5% yield). LC-MS, ES - : 576.57 [M-H] - .

[1007] Step 8: A solution of (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N- methyl-L-leucyl)-2-oxo-5-(propan-1-alkan-2-yl)spiro[indoline-3,3'-pyrrolidine]-5'- carboxamide (19 mg, 0.033 mmol) in DCM (0.4 ml) was treated dropwise with TEA (30 μl, 0.215 mmol) and TFAA (15 μl, 0.106 mmol) at 0 °C. The reaction was stirred at 0 °C for 30 minutes and then quenched with saturated sodium bicarbonate solution. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered and concentrated under vacuum. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-1-((3R,5'S)-5'-cyano-2-oxo-5-(propan-1-alkan-2-yl)spiro[indoline-3,3'- pyrrolidine]-1'-yl)-4-methyl-1-oxopropan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2- carboxamide (13 mg, 0.023 mmol, 70.6% yield) as a white solid. LC-MS, ES - : 558.36 [M-H] - ; 1 H NMR (400 MHz, Chloroform-d) δ 9.07 (s, 1H), 8.25 (s, 1H), 7.19 (dd, J = 8.2, 1.8 Hz, 1H), 6.96 (d, J = 1.8 Hz, 1H), 6.91 - 6.73 (m, 3H), 6.63 (td, J = 10.0, 1.9 Hz, 1H), 5.38 (dd, J = 9.0, 6.2 Hz, 1H), 5.09 (dd, J = 17.1, 8.6 Hz, 2H), 4.91 (t, J = 1.5 Hz, 1H), 4.51 (d, J = 10.5 Hz, 1H), 3.98 (d, J = 10.5 Hz, 1H), 3.46 (s, 3H), 2.89 (dd, J = 13.2, 8.9 Hz, 1H), 2.66 - 2.50 (m, 1H), 1.98 - 1.75 (m, 5H), 1.01 (d, J = 6.6 Hz, 3H), 0.96 (d, J = 6.5 Hz, 3H).

[1008] Example 49

[1009]

[1010] Step 1 : A solution of N-((S)-1-((3R,5'S)-5'-cyano-2-oxo-5-(propan-1-yl-2-yl)spiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-methyl-1-oxopropan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2-carboxamide (8 mg, 0.014 mmol) in THF (0.2 ml) and water (0.1 ml) was treated with potassium osmate (3.4 mg, 9.23 μmol) in water (0.1 ml). The reaction mixture was stirred at room temperature for 3 h, then quenched with saturated aqueous sodium thiosulfate solution. The resulting mixture was stirred at room temperature for an additional 30 min. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was loaded onto a 4 g silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2-carboxamide (6 mg, 10.68 μmol, 74.7% yield) as a white solid. LC-MS, ES - : 560.37 [M-H] - ; 1 H NMR (400 MHz, Chloroform-d) δ 9.73 (s, 1H), 8.23 (s, 1H), 7.77 (d, J = 1.7 Hz, 1H), 7.68 (dd, J = 8.2, 1.7 Hz, 1H), 6.92 (dd, J = 8.4, 2.2 Hz, 2H), 6.84 - 6.76 (m, 1H), 6.62 (td, J = 10.0, 2.0 Hz, 1H), 5.24 (dd, J = 8.4, 6.8 Hz, 1H), 5.12 (t, J = 8.3 Hz, 1H), 4.75 (d, J = 10.6 Hz, 1H), 3.99 (d, J = 10.5 Hz, 1H), 3.51 (s, 3H), 2.89 (dd, J = 13.4, 8.3 Hz, 1H), 2.54 (dd, J = 13.5, 8.3 Hz, 1H), 2.38 (s, 3H), 1.98 - 1.79 (m, 2H), 1.70 - 1.62 (m, 1H), 1.02 (d, J = 6.6 Hz, 3H), 0.97 (d, J = 6.5 Hz, 3H), 0.86 (d, J = 14.2 Hz, 1H).

[1011] Example 500

[1012]

[1013] Step 1 : A solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (43 mg, 0.070 mmol) in DMSO (0.5 ml) was treated with copper(I) iodide (3 mg, 0.016 mmol), sodium L-proline (6 mg, 0.044 mmol) and sodium methane sulfonate (13 mg, 0.127 mmol) under nitrogen. The resulting reaction mixture was bubbled with nitrogen for 5 minutes. The reaction mixture was warmed to 90 °C and stirred overnight. The reaction mixture was concentrated by a V10 evaporator. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-5- (methylsulfonyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (16 mg, 0.026 mmol, 37.3% yield) as a white solid. LC-MS, ES - : 614.24 [M-H] - .

[1014] Step 2: A solution of (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N- methyl-L-leucyl)-5-(methylsulfonyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'- carboxamide (15 mg, 0.024 mmol) in DCM (0.04 ml) was treated dropwise with TEA (30 μl, 0.215 mmol) and TFAA (11 μl, 0.078 mmol) at 0 °C. The reaction was stirred at 0 °C for 30 minutes then quenched with a saturated solution of sodium bicarbonate and the aqueous layer extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered and concentrated under vacuum. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-l-((3R,5'S)-5'-cyano-5-(methylsulfonyl)-2-oxospiro[indoline-3,3'- pyrrolidine]-l'-yl)-4-methyl-l-oxopentan-2-yl)-4,6-difluoro-N-methyl-lH-indole-2- carboxamide (12 mg, 0.020 mmol, 82% yield) as a white solid. LC-MS, ES - : 596.31 [M-H] - ; 1H NMR (500 MHz, Methanol-d4) δ 7.80 (dd, J = 8.3, 1.8 Hz, 1H), 7.64 (d, J = 1.9 Hz, 1H), 7.10 (d, J = 8.2 Hz, 1H), 6.98 - 6.89 (m, 2H), 6.65 (td, J = 10.2, 2.0 Hz, 1H), 5.38 (dd, J = 8.9, 6.3 Hz, 1H), 5.26 (t, J = 8.0 Hz, 1H), 4.36 (d, J = 10.9 Hz, 1H), 4.02 (d, J = 10.8 Hz, 1H), 3.44 (s, 3H), 2.95 (s, 3H), 2.74 (d, J = 8.1 Hz, 1H), 1.92 (ddd, J = 14.4, 8.9, 5.6 Hz, 1H), 1.82 (ddd, J = 14.2, 8.1, 6.2 Hz, 1H), 1.65 (ddd, J = 12.2, 7.9, 6.1 Hz, 1H), 1.34 - 1.22 (m, 2H), 1.04 (d, J = 6.7 Hz, 3H), 0.99 (d, J = 6.6 Hz, 3H), 0.93 - 0.85 (m, 1H).

[1015] Example 501

[1016]

[1017] Step 1 : Treat a solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (137 mg, 0.222 mmol) in DMF (0.5 mL) and MeOH (0.5 mL) with TEA (100 μL, 0.717 mmol), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (Xantphos) (20 mg, 0.035 mmol) and palladium(II) acetate (6.8 mg, 0.030 mmol) in a carbon monoxide environment (6.22 mg, 0.222 mmol) (1 atm). Bubble the resulting reaction mixture with carbon monoxide (6.22 mg, 0.222 mmol) for 5 minutes. Warm the reaction to 70 °C and stir overnight. Concentrate the reaction mixture in vacuo. Load the crude product onto a 4 gram silica gel column and elute with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-5'-carbamoyl-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5-carboxylic acid methyl ester (85 mg, 0.143 mmol, 64.2% yield) off-white solid. LC-MS, ES - 594.26 [M-H]- .

[1018] Step 2: A solution of (3R,5'S)-5'-carbamoyl-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5-carboxylic acid methyl ester (8 mg, 0.013 mmol) in DCM (0.3 mL) was treated dropwise with TEA (20 μL, 0.143 mmol) and TFAA (7 μL, 0.050 mmol) at 0 °C. The reaction was stirred at 0 °C for 30 min, then quenched with saturated sodium bicarbonate solution. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was loaded onto a 4 g silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield (3R,5'S)-5'-cyano-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5-carboxylic acid methyl ester (7 mg, 0.012 mmol, 90% yield) as a white solid. LC-MS, ES - : 576.34 [M-H] - ; 1 H NMR (500 MHz, Chloroform-d) δ 9.61 (s, 1H), 8.03 (s, 1H), 7.80 (dd, J = 8.1, 1.7 Hz, 1H), 7.76 (d, J = 1.7 Hz, 1H), 6.91 (d, J = 8.1 Hz, 1H), 6.89 - 6.82 (m, 2H), 6.63 (td, J = 10.0, 2.0 Hz, 1H), 5.25 (dd, J = 8.4, 6.9 Hz, 1H), 5.10 (t, J = 8.4 Hz, 1H), 4.78 (d, J = 10.5 Hz, 1H), 3.99 (d, J = 10.5 Hz, 1H), 3.80 (s, 3H), 3.54 (s, 3H), 2.90 (dd, J = 13.4, 8.5 Hz, 1H), 2.55 (ddd, J = 13.5, 8.4, 1.2 Hz, 1H), 1.93 (ddd, J = 14.3, 8.4, 6.1 Hz, 1H), 1.84 (dt, J = 14.2, 7.3 Hz, 1H), 1.64 (dt, J = 13.7, 6.8 Hz, 2H), 1.03 (d, J = 6.6 Hz, 3H), 0.97 (d, J = 6.6 Hz, 3H), 0.90 - 0.79 (m, 1H).

[1019] Example 50

[1020]

[1021] Step 1 : A solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (21 mg, 0.034 mmol) in 1,4-dioxane (0.4 ml) was treated with Pd(OAc)2(3 mg, 0.013 mmol), 4,5-bis diphenylphosphino-9,9-dimethylxanthene (Xantphos) (8 mg, 0.014 mmol), Co2(CO)8(8 mg, 0.023 mmol), DMAP (9 mg, 0.074 mmol) and morpholine (10 μl, 0.115 mmol) under nitrogen. The resulting reaction mixture was bubbled with nitrogen for 3 minutes. The reaction was warmed to 90 °C and stirred under microwave irradiation for 30 minutes. Upon completion of the reaction, the mixture turned to a black suspension. The reaction mixture was filtered through celite and rinsed three times with acetone. The filtrate was concentrated in vacuo. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-5-(morpholine-4-carbonyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (15 mg, 0.023 mmol, 67.7% yield) as a bright yellow solid. LC-MS, ES - : 649.43 [M-H] - .

[1022] Step 2: A solution of (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N- methyl-L-leucyl)-5-(morpholine-4-carbonyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'- carboxamide (5 mg, 7.68 μmol) in THF (0.3 ml) was treated with Burgess reagent (11 mg, 0.046 mmol). The reaction mixture was stirred at room temperature for 3 h. The crude product mixture was loaded onto a 4 gram silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-l-((3R,5'S)-5'-cyano-5-(morpholine-4-carbonyl)-2-oxospiro[indoline-3,3'- pyrrolidine]-l'-yl)-4-methyl-l-oxopentan-2-yl)-4,6-difluoro-N-methyl-lH-indole-2- carboxamide (4 mg, 6.32 μmol, 82% yield) as a white solid. LC-MS, ES - : 631.26 [M-H] - ; 1H NMR (400 MHz, Chloroform-d) δ 11.35 (s, 1H), 8.26 (s, 1H), 7.25 (d, J = 1.7 Hz, 1H), 7.13 (dd, J = 8.0, 1.6 Hz, 1H), 6.98 (dd, J = 9.8, 2.3 Hz, 1H), 6.89 - 6.83 (m, 1H), 6.71 (d, J = 8.0 Hz, 1H), 6.60 (td, J = 10.0, 2.0 Hz, 1H), 5.05 - 4.88 (m, 2H), 4.82 (d, J = 10.2 Hz, 1H), 3.98 (d, J = 10.1 Hz, 2H), 3.89 - 3.64 (m, 5H), 3.58 (s, 3H), 3.54 - 3.32 (m, 3H), 2.89 (dd, J = 13.1, 10.4 Hz, 1H), 2.55 - 2.41 (m, 1H), 2.18 (d, J = 2.3 Hz, 1H), 1.92 (dq, J = 17.2, 6.7 Hz, 2H), 1.71 (p, J = 6.7 Hz, 2H), 1.09 (d, J = 6.6 Hz, 3H), 1.03 (d, J = 6.5 Hz, 3H), 0.84 (s, 1H).

[1023] Example 50

[1024]

[1025] Step 1 : A solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (65 mg, 0.105 mmol) in ethanol (0.3 mL) and water (0.15 mL) was treated with sodium azide (18 mg, 0.277 mmol), copper(I) iodide (4.6 mg, 0.024 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (7 μL, 0.044 mmol), and sodium ascorbate (5.4 mg, 0.027 mmol) under a nitrogen atmosphere. The resulting reaction mixture was bubbled with nitrogen for 5 minutes. The reaction was heated to 100 °C and stirred under microwave irradiation for 30 minutes. The reaction mixture was concentrated in vacuo. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-5-azido-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (33 mg, 0.057 mmol, 54.1% yield) as a yellow solid. LC-MS, ES - : 577.36 [M-H] -.

[1026] Step 2: A solution of (3R,5'S)-5-azido-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (33 mg, 0.057 mmol) in DCM (0.5 ml) was treated dropwise with TEA (50 μl, 0.359 mmol) and TFAA (25 μl, 0.177 mmol) at 0 °C. The reaction was stirred at 0 °C for 30 minutes and then quenched with saturated sodium bicarbonate solution. The aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered and concentrated under vacuum. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-l-((3R,5'S)-5-azido-5'-cyano-2-oxospiro[indoline-3,3'- pyrrolidine]-l'-yl)-4-methyl-l-oxopentan-2-yl)-4,6-difluoro-N-methyl-lH-indole-2- carboxamide (14 mg, 0.025 mmol, 43.8% yield) as a white solid. LC-MS, ES - : 559.34 [M-H] - ; 1 H NMR (400 MHz, Chloroform-d) δ 9.17 (s, 1H), 8.27 (s, 1H), 6.93 (s, 1H), 6.83 (dd, J = 8.9, 3.0 Hz, 2H), 6.71 - 6.55 (m, 2H), 6.42 (d, J = 2.2 Hz, 1H), 5.39 (dd, J = 9.2, 6.0 Hz, 1H), 5.03 (t, J = 8.4 Hz, 1H), 4.55 (d, J = 10.6 Hz, 1H), 3.95 (d, J = 10.6 Hz, 1H), 3.50 (s, 3H), 2.88 (dd, J = 13.4, 8.5 Hz, 1H), 2.53 (dd, J = 13.4, 8.5 Hz, 1H), 2.18 (d, J = 2.5 Hz, 1H), 1.95 (ddd, J = 14.4, 9.1, 5.4 Hz, 1H), 1.79 (ddd, J = 14.2, 8.4, 6.0 Hz, 1H), 1.02 (d, J = 6.6 Hz, 3H), 0.96 (d, J = 6.5 Hz, 3H), 0.87 (d, J = 11.0 Hz, 1H).

[1027] Example 50

[1028]

[1029] Step 1 : A solution of (3R,5'S)-5-bromo-l'-(N-(4,6-difluoro-lH-indole-2- carbonyl)-N-methyl-L-leucinyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (65 mg, 0.105 mmol) in ethanol (0.3 mL) and water (0.15 mL) was treated with sodium azide (18 mg, 0.277 mmol), copper(I) iodide (4.6 mg, 0.024 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (7 μL, 0.044 mmol), and sodium ascorbate (5.4 mg, 0.027 mmol) under a nitrogen atmosphere. The resulting reaction mixture was sparged with nitrogen for 5 minutes. The reaction was heated to 100 °C and stirred under microwave irradiation for 30 minutes. The reaction mixture was concentrated in vacuo. The crude product was loaded onto a 4 gram silica gel cartridge and eluted with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-5-amino-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucinyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide as a yellow solid (19 mg, 0.034 mmol, 32.6% yield). LC-MS, ES - : 551.27 [M-H] - .

[1030] Step 2: A solution of (3R,5'S)-5-amino-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N- methyl-L-leucinyl)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'-carboxamide (15 mg, 0.027 mmol) and 1-fluorocyclopropyl-1-carboxylic acid (4.7 mg, 0.045 mmol) in DMF (0.1 mL) and DCM (0.3 mL) was treated with HATU (15 mg, 0.039 mmol) and N-methylmorpholine (20 μL, 0.182 mmol) at room temperature. The reaction mixture was stirred for 3 h, then quenched with a saturated solution of sodium bicarbonate. The aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was loaded onto a 4 gram silica gel cartridge and eluted with 0% to 100% acetone / cyclohexane to yield (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N-methyl-L-leucinyl)-5-(l- fluorocyclopropyl-1-carboxylic acid amide)-2-oxospiro[indoline-3,3'-pyrrolidine]-5'- carboxamide as a bright yellow solid (15 mg, 0.023 mmol, 87% yield). LC-MS, ES - : 637.69 [M-H] - .

[1031] Step 3: A solution of (3R,5'S)-l'-(N-(4,6-difluoro-lH-indole-2-carbonyl)-N- methyl-L-leucyl)-5-(l-fluorocyclopropyl-l-carboxyamide)-2-oxospiro[indoline-3,3'- pyrrolidin]-5'-carboxamide (15 mg, 0.023 mmol) in DCM (0.3 ml) was treated with Burgess reagent (17 mg, 0.071 mmol). The reaction mixture was stirred at room temperature for 3 h. The resulting mixture was loaded onto a 4 g silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-l-((3R,5'S)-5'-cyano-5-(l-fluorocyclopropyl-l- carboxyamide)-2-oxospiro[indoline-3,3'-pyrrolidin]-l'-yl)-4-methyl-l-oxopentan-2-yl)- 4,6-difluoro-N-methyl-lH-indole-2-carboxamide (12 mg, 0.019 mmol, 82% yield) as a white solid. LC-MS, ES - : 619.25 [M-H] - ; 1 H NMR (500 MHz, Chloroform-d) δ 9.43 (s, 1H), 8.28 (s, 1H), 8.20 (d, J = 5.1 Hz, 1H), 7.35 (dd, J = 8.4, 2.1 Hz, 1H), 7.25 (d, J = 2.2 Hz, 1H), 6.92 (d, J = 2.3 Hz, 1H), 6.79 (dd, J = 24.3, 8.5 Hz, 2H), 6.63 (td, J = 10.0, 2.0 Hz, 1H), 5.20 (dd, J = 10.0, 5.2 Hz, 1H), 5.04 (t, J = 8.6 Hz, 1H), 4.42 (d, J = 10.6 Hz, 1H), 3.98 (d, J = 10.5 Hz, 1H), 3.52 (s, 3H), 2.88 (dd, J = 13.2, 9.0 Hz, 1H), 2.61 - 2.49 (m, 1H), 1.99 (ddd, J = 14.4, 10.1, 4.8 Hz, 1H), 1.81 - 1.56 (m, 4H), 1.52 - 1.34 (m, 4H), 1.03 (d, J = 6.4 Hz, 3H), 0.94 (d, J = 6.3 Hz, 3H), 0.78 - 0.90 (m, 1H).

[1032] Example 505

[1033]

[1034] Step 1 : A solution of N-((S)-1-((3R,5'S)-5-azido-5'-cyano-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2- carboxamide (8 mg, 0.014 mmol) and ethyl cyclopropyl (3 μΐ, 0.035 mmol) in tBuOH (0.2 ml) and water (0.2 ml) was treated with copper (II) sulfate pentahydrate (1.7 mg, 6.81 μmol) and sodium ascorbate (3.3 mg, 0.017 mmol). The resulting reaction mixture was stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate solution. The aqueous layer was extracted with dichloromethane three times. The combined organic layers were dried over sodium sulfate, filtered and concentrated under vacuum. The crude product was loaded onto a 4 gram silica gel column and eluted with 0% to 100% ethyl acetate / cyclohexane to yield N-((S)-1-((3R,5'S)-5'-cyano-5-(4-cyclopropyl-1H-1,2,3-triazol-1-yl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)-4,6-difluoro-N-methyl-1H-indole-2- carboxamide (2.4 mg, 3.83 μmol, 26.8% yield) as a white solid. LC-MS, ES + : 627.53 [M+H] + ; 1 HNMR (500 MHz, Chloroform-d) δ 10.34 (s, 1H), 7.86 (s, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.54 (s, 1H), 7.32 (dt, J = 9.2, 2.6 Hz, 2H), 7.00 (d, J = 8.3 Hz, 1H), 6.83 (d, J = 2.2 Hz, 1H), 6.62 (td, J = 10.0, 2.1 Hz, 1H), 5.00 (ddd, J = 20.4, 8.9, 7.3 Hz, 2H), 4.85 (d, J = 10.3 Hz, 1H), 4.04 (d, J = 10.3 Hz, 1H), 3.56 (s, 3H), 2.94 (dd, J = 13.3, 9.4 Hz, 1H), 2.57 (dd, J = 13.2, 8.0 Hz, 1H), 2.18 (d, J = 2.9 Hz, 2H), 2.08 - 1.94 (m, 2H), 1.80 (dt, J = 14.1, 7.1 Hz, 1H), 1.70 (dt, J = 13.5, 6.7 Hz, 1H), 1.26 (s, 2H), 1.05 (dd, J = 7.6, 4.5 Hz, 4H), 1.02 - 0.95 (m, 3H), 0.95 - 0.87 (m, 3H), 0.85 (d, J = 12.7 Hz, 1H).

[1035] Example 506

[1036]

[1037] Step 1 : To a solution of the bromide (85 mg, 0.138 mmol) and pyridine-3- ylboronic acid (25 mg, 0.207 mmol) in THF (0.7 mL) was added XPhos Pd G3 (12 mg, 0.014 mmol) and K3PO4 (0.55 mL, 0.5 M aq). The reaction mixture was heated to 85 °C for 18 h. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated. The residue was purified on a silica gel chromatography column with 0-80% acetone in cyclohexane to yield the desired product (25 mg, 30%).

[1038] Step 2: To a solution of the material from Step 1 (25 mg, 0.041 mmol) in DCM (1 mL) was added TFAA (17 μL, 0.122 mmol) and Et3N (34 μL, 0.244 mmol) at 0 °C. The crude product was directly loaded on a silica gel column and chromatographed with 0-80% acetone in cyclohexane to yield EP-040278 (15 mg, 62%). 595.343 [M-H].1H NMR (400 MHz, Acetone-d6) δ 11.07 (s, 1H), 9.89 (s, 1H), 9.02 (d, J = 2.5 Hz, 1H), 8.57 (dd, J = 4.8, 1.6 Hz, 1H), 7.72 (dt, J = 8.1, 1.9 Hz, 1H), 7.48 (dd, J = 8.1, 1.9 Hz, 1H), 7.37 (d, J = 1.9 Hz, 1H), 7.26 (dd, J = 8.0, 4.8 Hz, 1H), 7.21 - 7.15 (m, 1H), 7.15 - 7.07 (m, 1H), 6.86 (dd, J = 2.5, 0.9 Hz, 1H), 6.77 (td, J = 10.3, 2.1 Hz, 1H), 5.56 (dd, J = 10.7, 4.6 Hz, 1H), 5.37 (t, J = 8.5 Hz, 1H), 4.53 (d, J = 10.8 Hz, 1H), 4.06 (d, J = 10.7 Hz, 1H), 3.39 (s, 3H), 2.83 (td, J = 6.4, 3.0 Hz, 5H), 2.75 (dd, J = 13.2, 8.6 Hz, 1H), 2.12 - 1.92 (m, 5H), 1.83 - 1.57 (m, 2H), 1.00 (dd, J = 26.6, 6.3 Hz, 6H).

[1039] Example 507

[1040]

[1041]

[1042] Step 1 : To a solution of the bromide (252 mg, 0.409 mmol) in dioxane (2 mL) was added ethynyltrimethylsilane (87 μL, 0.613 mmol), Pd(PPh3)Cl2(28.7 mg, 0.041 mmol), CuI (15.6 mg, 0.082 mmol), and Et3N (171 μL, 1.23 mmol). The reaction mixture was heated to 85 °C for 18 h. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on silica gel chromatography with 0-80% acetone in cyclohexane to give the desired product (165 mg, 64%).

[1043] Step 2: To a solution of the material from Step 1 (139 mg, 0.219 mmol) in methanol (2 mL) was added K2CO3(61 mg, 0.439 mmol). The resulting reaction mixture was stirred at room temperature for 18 h. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on silica gel chromatography with 0-80% acetone in cyclohexane to give the desired product (77 mg, 63%).

[1044] Step 3: To a solution of the material from Step 2 (77 mg, 0.137 mmol) and tetrazolo[l,5-a]pyridine (25 mg, 0.206 mmol) in DMF (2 mL) was added CuSO4-pentahydrate (9 mg, 0.036 mmol), sodium ascorbate (7 mg, 0.035 mmol), and water (1 mL). The reaction mixture was heated to 80 °C for 18 h. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated. The residue was purified on silica gel chromatography with 0-80% acetone in cyclohexane to give the desired product (50 mg, 54%).

[1045] Step 4: To a solution of the material from Step 3 (50 mg, 0.041 mmol) in DCM (1 mL) at 0 °C was added TFAA (31 μL, 0.220 mmol) and Et3N (62 μL, 0.440 mmol). The crude product was loaded directly on a silica gel column and chromatographed with 0-80% acetone in cyclohexane to yield EP-040469.662.405 [M-H].1H NMR (400 MHz, Acetone-d6) δ 10.49 (s, 1H), 9.87 (s, 1H), 8.58-8.49 (m, 2H), 8.18-8.08 (m, 2H), 7.71 (d, J = 8.2 Hz, 1H), 7.67 (d, J = 1.7 Hz, 1H), 7.52 (ddd, J = 6.3, 4.8, 2.2 Hz, 1H), 7.05 (d, J = 8.0 Hz, 1H), 7.03-6.94 (m, 1H), 6.70 (dd, J = 2.3, 1.0 Hz, 1H), 6.44 (td, J = 10.3, 2.1 Hz, 1H), 5.67-5.59 (m, 1H), 5.40 (t, J = 8.4 Hz, 1H), 4.71 (d, J = 10.7 Hz, 1H), 4.00 (d, J = 10.8 Hz, 1H), 3.53 (s, 3H), 3.25 (s, 1H), 2.77 (dd, J = 13.3, 8.1 Hz, 1H), 2.09-2.04 (m, 2H), 1.98 (ddd, J = 14.3, 9.4, 5.2 Hz, 1H), 1.79 (ddd, J = 14.1, 8.6, 5.7 Hz, 1H), 1.62 (dt, J = 14.4, 6.9 Hz, 1H), 1.44 (d, J = 0.9 Hz, 2H), 1.31 (s, 2H), 0.98 (dd, J = 22.7, 6.5 Hz, 7H).

[1046] The following examples were prepared using similar methods as described above.

[1047]

[1048]

[1049]

[1050]

[1051]

[1052]

[1053]

[1054] Example 517

[1055]

[1056] Step 1 1 '-tert-butyl 5'-methyl (3R,5'S)-5-bromo-2-oxospiro[indoline-3,3'- pyrrolidin]-1 ',5'-carbonate (897 mg, 2.11 mmol) was dissolved in MTBE (14 mL) and cooled to 0 °C. Lithium borohydride (2.0 M in THF, 5.3 mL, 5 eq.) was added slowly and the resulting reaction mixture was stirred at 0 °C. After 40 min, an additional 2.5 mL of lithium borohydride solution was added and after 60 min, an additional 2.5 mL was added. The reaction was quenched with saturated ammonium chloride. The reaction mixture was extracted with ethyl acetate and diethyl ether, the collected organic portion was washed with brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was loaded onto preparative HPLC to yield tert-butyl (3R,5'S)-5-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-carboxylate (202 mg, 0.508 mmol, 24% yield).

[1057] Step 2

[1058] Tert-butyl (3R,5'S)-5-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-carboxylate (60 mg, 0.151 mmol) was dissolved in DCM / TFA (1 mL, 1 : 1 ratio) and stirred at room temperature. After 45 min, the reaction mixture was directly concentrated. The residue was re-dissolved in methanol and re-concentrated. The residue was then re-dissolved in ethyl acetate and re-concentrated. The resulting residue was used directly without further purification.

[1059] Step 3

[1060] Crude (3R,5'S)-5-bromo-5'-(hydroxymethyl)spiro[indoline-3,3'-pyrrolidin]-2-one 2,2,2-trifluoroacetate (crude, assumed 0.161 mmol) was dissolved in DMF (0.644 mL, 0.25 M) followed by the addition of iPr2NEt (87 mg, 0.676 mmol, 4.2 eq.) and N-((benzyloxy)carbonyl)-N-methyl-L-leucine (54 mg, 0.193 mmol, 1.2 eq.). Once a homogeneous solution was obtained, HATU (73 mg, 0.193 mmol, 1.2 eq.) was added. After 16 h, the reaction mixture was diluted with water and the mixture was subjected to solid phase extraction on an Oasis HLB (200 mg) extraction cartridge eluted with water and methanol to yield benzyl ((S)-1-((3R,5'S)-5-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (94 mg).

[1061] Step 4

[1062] Benzyl ((S)-1-((3R,5'S)-5-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)carbamate (crude, assumed 0.173 mmol) was dissolved in CH2Cl2(1.5 ml) and cooled to 0 °C. Dess-Martin periodinane (110 mg, 0.259 mmol) was added. The reaction mixture was stirred at 0 °C for 2 h, then TLC showed complete consumption of starting material (1 : 1 hexanes s / ethyl acetate). The reaction was then partitioned between DCM and saturated Na2S2O3, followed by washing of the organic layer with saturated sodium bicarbonate and brine, drying over magnesium sulfate, filtration, and concentration. The crude product residue was used without further purification.

[1063] Step 5

[1064] Benzyl ((S)-1-((3R,5'S)-5-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)carbamate (crude, assumed 0.173 mmol) was dissolved in CH2Cl2(6 mL) followed by the addition of iPr2NEt (67 mg, 0.519 mmol, 3 eq.) and hydroxylamine hydrochloride (240 mg, 3.46 mmol). After 4 days, ethyl acetate and water were added. The organic phase was washed with brine and dried over magnesium sulfate, filtered, and concentrated. The crude product residue was used without further purification.

[1065] Step 6

[1066] Benzyl ((S)-1-((3R,5'S)-5-bromo-5'-((hydroxyimino)methyl)-2-oxospiro[indoline- 3,3'-pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (assumed 0.161 mmol) was dissolved in MeCN (3.22 mL) and copper(II) acetate (9 mg, 0.05 mmol) was added. The reaction mixture was heated to 70 °C for 80 min. Purification of the mixture with preparative HPLC yielded benzyl ((S)-1-((3R,5'S)-5-bromo-5'-cyano-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (1.1 mg, 0.002 mmol, 1% yield over 5 steps).1H NMR (400 MHz, Methanol-d4) δ 7.40 (dd, J = 8.3, 2.0 Hz, 1H), 7.35 - 7.15 (m, 4H), 7.13 - 7.05 (m, 2H), 6.89 (dd, J = 8.4, 3.3 Hz, 1H), 5.18 (t, J = 8.2 Hz, 1H), 4.99 (dd, J = 9.1, 6.1 Hz, 1H), 4.95 - 4.89 (m, 1H), 4.76 (d, J = 12.3 Hz, 1H), 4.22 (d, J = 10.9 Hz, 1H), 3.87 (d, J = 10.8 Hz, 1H), 1.77 (ddd, J = 14.3, 9.0, 5.4 Hz, 1H), 1.65 (ddd, J = 13.9, 8.2, 5.9 Hz, 1H), 1.48 (dt, J = 13.9, 6.8 Hz, 1H), 0.94 (dd, J = 15.3, 6.5 Hz, 6H). [M+Na] m / z 574.82.

[1067] Example 518

[1068]

[1069]

[1070] Step 1

[1071] Tert-butyl (3R,5'S)-5'-bromo-5'-(hydroxymethyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-carboxylate (60 mg, 0.151 mmol), cesium carbonate (148 mg, 0.453 mmol), potassium trifluoro(methyl)borate (36.8 mg, 0.302 mmol), and Pd(dppf)Cl2(27.6 mg, 0.038 mmol) were heated overnight at 80 °C in a sealed vessel under a nitrogen atmosphere. The reaction mixture was purified by preparative HPLC to yield tert-butyl (3R,5'S)-5'-(hydroxymethyl)-5-methyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-carboxylate (9 mg, 0.27 mmol, 18%).

[1072] Step 2

[1073] Tert-butyl (3R,5'S)-5'-(hydroxymethyl)-5-methyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-carboxylate (60 mg, 0.151 mmol) was dissolved in DCM / TFA (1 mL, 1 : 1 ratio) and stirred at room temperature. After 45 minutes, the reaction mixture was directly concentrated. The residue was again dissolved in methanol and concentrated again. The residue was then again dissolved in ethyl acetate and concentrated again. The resulting residue was used directly without further purification.

[1074] Step 3

[1075] Crude (3R,5'S)-5'-(hydroxymethyl)-5-methylspiro[indoline-3,3'-pyrrolidin]-2-one (crude, assumed 0.045 mmol) was dissolved in DMF (0.563 mL, 0.08 M) followed by the addition of iPr2NEt (38 mg, 0.293 mmol, 6.5 eq.) and N-((benzyloxy)carbonyl)-N-methyl-L-leucine (38 mg, 0.135 mmol, 3 eq.). Once a homogeneous solution was obtained, HATU (51 mg, 0.135 mmol, 3 eq.) was added. After 16 hours, the reaction mixture was diluted with water and the mixture was subjected to solid phase extraction on an Oasis HLB (200 mg) extraction cartridge eluted with water and methanol to yield benzyl ((S)-1-((3R,5'S)-5'-(hydroxymethyl)-5-methyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1 '-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (42 mg). [M+H] m / z 494.387.

[1076] Step 4

[1077] Benzyl ((S)-1-((3R,5'S)-5'-hydroxymethyl-5-methyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)carbamate (42 mg, assumed 0.088 mmol) was dissolved in CH2Cl2(1.5 mL) and cooled to 0 °C. Then Dess-Martin Oxidizing reagent (110 mg, 0.259 mmol) was added. The reaction mixture was stirred at 0 °C for 2 h, then TLC indicated consumption of starting material (1 : 1 hexanes / ethyl acetate). The reaction mixture was then partitioned between DCM and saturated Na2S2O3, the combined organic phases were washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, filtered, and concentrated. The crude product residue was used without further purification.

[1078] Step 5

[1079] Benzyl ((S)-1-((3R,5'S)-5'-formyl-5-methyl-2-oxospiro[indoline-3,3'-pyrrolidin]-1'- yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (crude, assumed 0.088 mmol) was dissolved in CH2Cl2(4 mL), then iPr2NEt (34 mg, 0.264 mmol) and hydroxylamine hydrochloride (153 mg, 2.20 mmol) were added. After 4 days, EtOAc and water were added. The organic layer was washed with brine, then dried over magnesium sulfate, filtered, and concentrated. The crude product residue was used without further purification.

[1080] Step 6

[1081] Benzyl ((S)-1-((3R,5'S)-5'-((hydroxyimino)methyl)-5-methyl-2-oxospiro[indoline- 3,3'-pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (crude, assumed 0.045 mmol) was dissolved in MeCN (3 mL) and copper(II) acetate (9 mg, 0.05 mmol) was added. The reaction mixture was heated to 70 °C for 80 min. The reaction mixture was worked up with preparative HPLC to yield benzyl ((S)-1-((3R,5'S)-5'-cyano-5-methyl-2-oxospiro[indoline-3,3'- pyrrolidin]-1'-yl)-4-methyl-1-oxopentan-2-yl)(methyl)carbamate (1.04 mg, 0.002 mmol, 5% yield over five steps).1H NMR (400 MHz, Acetone-d6) δ 9.60 (s, 1H), 7.43 - 7.17 (m, 5H), 7.15 - 7.04 (m, 1H), 6.92 (d, J = 7.8 Hz, 1H), 6.81 (t, J = 1.3 Hz, 1H), 5.22 - 5.14 (m, 1H), 5.08 (dd, J = 9.6, 5.6 Hz, 1H), 4.94 (d, J = 12.6 Hz, 1H), 4.67 (d, J = 12.6 Hz, 1H), 4.35 - 4.25 (m, 1H), 3.91 (dd, J = 10.4, 4.1 Hz, 1H), 2.94 (s, 3H), 2.22 (s, 3H), 1.81 (ddd, J = 14.3, 9.6, 5.0 Hz, 1H), 1.64 (ddd, J = 14.1, 8.7, 5.7 Hz, 1H), 1.52 (p, J = 6.4 Hz, 1H), 1.04 - 0.91 (m, 6H). [M+Na] m / z 511.119.

[1082] Example 519

[1083]

[1084] (S)-3-cyclopropyl-2-(4-fluoro-3-methylbenzo[b]thiophene-2-carboxylic acid amide)propionic acid (33 mg, 0.103 mmol) and (3R,5'S)-5-chloro-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-carboxamide hydrochloride (27 mg, 0.089 mmol) were dissolved in DMF (0.40 mL). HATU (39 mg, 0.103 mmol) and iPr2NEt (0.041 mL, 0.232 mmol) were then added sequentially and the resulting reaction solution was stirred at room temperature for 20 minutes. Additional HATU (-10 mg) and iPr2NEt (2-3 drops) were added as needed (in this case, after 20 minutes). After 1 hour at room temperature, palladium (II) trifluoroacetate (30 mg, 0.089 mmol), dichloroacetonitrile (0.4 mL), and water (0.4 mL) were added sequentially. The resulting suspension was heated to 65 °C for 1 hour and then cooled to room temperature. The reaction mixture was filtered and treated on a preparative HPLC to yield N-((S)-l-((3R,5'S)-5-chloro-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-l'-yl)-3-cyclopropyl-l- oxoprop-2-yl)-4-fluoro-3-methylbenzo[b]thiophene-2-carboxamide (1.20 mg, 0.0022 mmol, 2% yield).1H NMR (500 MHz, Acetone-d6) δ 9.82 (s, 1H), 7.80-7.70 (m, 2H), 7.48 (td, J = 8.0, 4.8 Hz, 1H), 7.36-7.25 (m, 2H), 7.16 (ddd, J = 12.2, 8.0, 0.8 Hz, 1H), 7.02 (d, J = 8.2 Hz, 1H), 5.29 (t, J = 8.2 Hz, 1H), 4.88 (q, J = 7.0 Hz, 1H), 4.50-4.44 (m, 1H), 4.13 (d, J = 10.5 Hz, 1H), 2.86 (ddd, J = 13.3, 8.5, 1.1 Hz, 1H), 2.80 (s, 3H), 2.73 (dd, J = 13.3, 7.9 Hz, 1H), 1.85 (t, J = 7.0 Hz, 2H), 0.95 (dddd, J = 15.1, 10.2, 5.2, 2.4 Hz, 1H), 0.58-0.52 (m, 2H), 0.30 (dddd, J = 9.4, 4.7, 2.7, 1.4 Hz, 1H), 0.24 (dddd, J = 9.0, 6.3, 2.8, 1.2 Hz, 1H). [M+H] + 550.741

[1085] The following examples were prepared using similar procedures as described above.

[1086]

[1087]

[1088]

[1089]

[1090]

[1091] Example 527

[1092]

[1093] iPr2NEt (35.0 μl, 0.200 mmol) was added to a stirred solution of 2,5-dioxopyrrolidin-l- yl(((S)-2,2,2-trifluoro-l-phenylethoxy)carbonyl)-L-leucinate (66.3 mg, 0.154 mmol), (3R,5'S)- 5-fluoro-2-oxospiro[indoline-3,3'-pyrrolidin]-5'-formamide hydrochloride (44 mg, 0.154 mmol), and MeCN (0.380 ml). The resulting reaction mixture was stirred at room temperature for 3 h, then warmed to 50 °C and maintained for 16 h. After this time, 15 u of LiPr2Net and 20 uL of 2,5-dioxopyrrolidin-l-yl(((S)-2,2,2-trifluoro-l- phenylethoxy)carbonyl)-L-leucinate were added, and the resulting mixture was further warmed to 65 °C. After 24 h, palladium (II) trifluoroacetate (51 mg, 0.154 mmol), dichloroacetonitrile (0.38 mL), and water (0.38 mL) were added, and stirring was continued at 65 °C. After 25 min, the reaction mixture was cooled to room temperature, filtered, and worked up using preparative HPLC to yield (S)-2,2,2-trifluoro-l-phenylethyl ((S)-l-((3R,5'S)-5'-cyano-5-fluoro-2-oxospiro[indoline-3,3'- pyrrolidin]-l'-yl)-4-methyl-l-oxopentan-2-yl)carbamate (0.96 mg, 0.0017 mmol, 1% yield).1H NMR (500 MHz, Acetone-d6) δ 9.70 (s, 1H), 7.52 (dt, J = 6.6, 3.8 Hz, 2H), 7.46 (dp, J = 4.6, 1.7 Hz, 3H), 7.32 (d, J = 8.0 Hz, 1H), 7.07 (ddd, J = 9.4, 8.6, 2.6 Hz, 1H), 7.03 - 6.95 (m, 2H), 6.09 (q, J = 7.1 Hz, 1H), 5.24 (t, J = 8.2 Hz, 1H), 4.43 (ddd, J = 9.7, 7.9, 4.6 Hz, 1H), 4.31 - 4.18 (m, 1H), 4.00 (d, J = 10.4 Hz, 1H), 2.82 (ddd, J = 13.3, 8.5, 1.0 Hz, 2H), 2.70 (dd, J = 13.3, 7.9 Hz, 1H), 1.83 - 1.59 (m, 3H), 0.95 (d, J = 6.6 Hz, 3H), 0.88 (d, J = 6.5 Hz, 3H). [M+H] + 547.20.

[1094] Example 528

[1095]

[1096] Step 1

[1097] A 37% aqueous solution of formaldehyde (0.312 mL, 4.19 mmol, 1.12 eq.) was added to a solution of (S)-2-amino-3-(4-chloro-lH-indol-3-yl)propanoic acid (892 mg, 3.74 mmol) and 0.5 N NaOH (1.1 eq. NaOH, 8.22 mL) at room temperature. The resulting reaction mixture was stirred at room temperature for 2 h, then at 104 °C for 24 h. During this time, additional formaldehyde solution was charged to the reaction mixture if necessary (0.3 mL was added after 3 h in this case). The reaction mixture was cooled to room temperature and acetic acid (0.449 mL, 7.85 mmol, 2.1 eq.) was added. The resulting mixture was then filtered, rinsing with water and THF sequentially to yield crude A-l, which was used without further purification.

[1098] Step 2

[1099] A-l (assumed 100% yield) was added to a 0.44 M solution of SOCl2in MeOH (15 mL, 0.25 M relative to substrate concentration). The resulting reaction mixture was stirred at 40 °C for 5 days. When judged complete by LCMS, the mixture was concentrated directly to yield crude B-l, which was used without further purification. [M+H] m / z 265.051.

[1100] Step 3.

[1101] Compound B-l (assumed 100% yield) was suspended in DCM (14 mL), then Hunig's base (1.25 mL, 7.2 mmol) was added, followed by a solution of di-tert-butyl dicarbonate (1.2 g, 5.5 mmol) in DCM (2.5 mL). When TLC (10:1 DCM / MeOH) indicated that the starting material was consumed, the reaction mixture was concentrated directly. The residue was chromatographed on silica gel, eluting with 0-35% ethyl acetate in hexanes to yield C-l (404 mg, 1.16 mmol, 31% yield over 3 steps).

[1102] Step 4

[1103] NBS (177 mg, 0.997 mmol, 0.9 eq.) was added to a vigorously stirred mixture of C-1 (404 mg, 1.107 mmol) in 2-MeTHF / water / acetic acid (63:28:9 ratio, 7.7 mL total volume, 0.114 M) at 0 °C. The resulting reaction mixture was stirred at 0 °C for 90 min, then slowly warmed to room temperature. When TLC showed the starting material was consumed, the reaction was quenched with saturated aqueous NaHC03, extracted with Et20. The organic phase was dried over MgS04, filtered, and concentrated. The residue was chromatographed on silica gel eluting with 0-50% ethyl acetate in hexanes to yield D-1 (251 mg, 0.66 mmol, 60% yield).

[1104] Step 5

[1105] D-1 (251 mg, 0.659 mmol) was dissolved in THF (3.30 ml, 0.2 M) and cooled to 0 °C. Lithium borohydride (2.0 M solution in THF, 1.98 mL, 6 eq.) was then added. When TLC (1 : 1 hexanes s / EtOAc) showed the starting material was completely consumed, the reaction was poured into an ice-cold saturated aqueous ammonium chloride solution. The reaction mixture was extracted with ethyl acetate and diethyl ether, the collected organic portions were washed with brine, dried over MgS04, filtered, and concentrated to yield crude product E-1 (228 mg), which was used without further purification.

[1106] Step 6

[1107] Crude product E-1 (50 mg, 0.142 mmol) was dissolved in DCM / TFA (4 mL, 1 : 1 ratio) and stirred at room temperature. After 45 min, the reaction mixture was directly concentrated. The residue was again dissolved in methanol and again concentrated. The residue was then again dissolved in ethyl acetate and again concentrated to yield F-1, which was used without further purification.

[1108] Step 7

[1109] Compound F-1 was dissolved in DMF (0.947 mL) and iPr2NEt (92 mg, 0.71 mmol), followed by the addition of (4-fluoro-lH-indole-2-carbonyl)-L-leucine (50 mg, 0.17 mmol). When the reaction was homogenous, HATU (65 mg, 0.17 mmol) was added. The resulting mixture was stirred at room temperature for 2.5 days, then diluted with water and DCM. The organic phase was washed with saturated aqueous NaHC03and brine, then dried over MgS04, filtered, and concentrated to yield crude product G-1 (assumed 100% yield). [M+H] m / z 526.913.

[1110] Step 8

[1111] The crude product G-1 was dissolved in CH2Cl2(1.42 mL) and cooled to 0 °C. Dess-Martin periodinane (110 mg) was added and the resulting mixture was stirred at 0 °C. After 1 h, Dess-Martin periodinane (80 mg) was added again. After 2 h, Dess-Martin periodinane (200 mg) was added again. The reaction mixture was then quenched with saturated aqueous Na2S2O3solution, and the resulting mixture was extracted with DCM. The organic phase was washed with saturated aqueous NaHCO3solution and brine, then dried over MgSO4, filtered, and concentrated.

[1112] The crude product obtained by the above procedure was dissolved in DCM (10 mL) and iPr2NEt (92 mg, 0.71 mmol) was added. Hydroxylamine hydrochloride (247 mg, 3.55 mmol) was then added, and the resulting mixture was stirred at room temperature for 16 h. Water was added, the phases were separated, and the organic phase was dried over MgSO4and concentrated. The crude product obtained by the above procedure was dissolved in MeCN (3 mL) and copper (II) acetate (9 mg, 0.05 mmol) was added. The resulting mixture was heated to 70 °C for 1 h. The resulting mixture was cooled to room temperature and treated with preparative HPLC to yield Example 528 (0.70 mg). 1 H NMR (500 MHz, Acetone-d6) δ 7.42 - 7.36 (m, 1H), 7.26 - 7.19 (m, 1H), 6.96 (m, 1H), 6.81 (m, 1H), 5.35 (m, 1H), 5.05 (m, 1H), 4.76 (mf, 1H), 4.69 (d, J = 11.0 Hz, 1H), 4.13 (d, J = 11.0 Hz, 1H), 3.12 (dd, J = 14.1, 9.7 Hz, 1H), 2.69 (dd, J = 14.1, 4.5 Hz, 1H), 1.83 (m, 2H), 1.78 - 1.68 (m, 1H), 1.06 - 0.93 (m, 6H). [M+H] m / z 521.835.

[1113] The following examples were prepared using similar methods as described above.

[1114]

[1115]

[1116]

[1117]

[1118] Example 535

[1119]

[1120] Step 1

[1121] (3R,5'S)-1'-((S)-4-fluoro-4-methyl-2-(methylamino)pentyl)-2-oxospiro[indoline-3,3'- pyrrolidin]-5'-carboxamide hydrochloride (198 mg, 0.48 mmol) was suspended in DCM (4.80 mL) and Et3N (334 uL, 2.40 mmol, 5 eq.) was added. The resulting mixture was cooled to 0 °C and acetyl chloride (42.9 uL, 0.528 mmol) was added dropwise. After 45 min at 0 °C, MeOH (1 mL) was added and the reaction mixture was directly concentrated. The residue was evaporated with ethyl acetate and used directly without further purification.

[1122] Step 2

[1123] The crude product from Step 1 (assuming 100% yield) was re-suspended in DCM (3.43 mL) and then Burgess reagent (0.343 g, 1.440 mmol) was added. The resulting mixture was stirred at room temperature overnight and then directly concentrated. The residue was chromatographed on silica gel eluting with 0-80% EtOAc in cyclohexane to yield N-((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-fluoro-4-methyl-1- oxopentan-2-yl)-N-methylacetamide (181 mg, 0.439 mmol, 91% yield over 2 steps).1H NMR (400 MHz, Acetone-d6) δ 9.67 (s, 1H), 7.22 (td, J = 7.6, 1.5 Hz, 1H), 7.03 - 6.91 (m, 3H), 6.48 (dd, J = 16.7, 10.4 Hz, 1H), 5.85 (dd, J = 16.7, 2.4 Hz, 1H), 5.70 (dd, J = 7.5, 5.7 Hz, 1H), 5.49 (dd, J = 10.4, 2.4 Hz, 1H), 5.16 (t, J = 8.3 Hz, 1H), 4.27 (dd, J = 10.7, 1.2 Hz, 1H), 3.89 (d, J = 10.7 Hz, 1H), 2.77 - 2.58 (m, 2H), 2.36 - 2.23 (m, 1H), 2.23 - 2.08 (m, 1H), 1.41 - 1.35 (m, 6H). [M+Na] m / z 435.33.

[1124] The following examples were prepared using similar methods as described above.

[1125]

[1126]

[1127]

[1128] Example 539

[1129]

[1130] Triethylamine (47 uL, 7 eq.) was added to a solution of N-((S)-1-((3R,5'S)-5'- cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-fluoro-4-methyl-1-oxopentan-2-yl)- N-methylacetamide (20 mg, 0.048 mmol) and dimethylamine hydrochloride (59 mg, 0.727 mmol) in MeOH (0.485 mL). When LCMS showed product formation, the reaction mixture was worked up using preparative HPLC to yield N-((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-4-fluoro-4-methyl-1- oxopentan-2-yl)-3-(dimethylamino)-N-methylpropan-1 -amine (1.95 mg, 0.0043 mmol, 9% yield).1H NMR (400 MHz, Acetone-d6) δ 7.32 (td, J = 7.7, 1.2 Hz, 1H), 7.12 (td, J = 7.5, 1.1 Hz, 1H), 7.04 (d, J = 7.7 Hz, 1H), 6.97 (dt, J = 7.4, 1.0 Hz, 1H), 5.68 (dd, J = 7.2, 6.0 Hz, 1H), 5.19 (t, J = 8.4 Hz, 1H), 4.28 (dd, J = 10.8, 1.3 Hz, 1H), 3.86 (d, J = 10.7 Hz, 1H), 3.01 (s, 3H), 2.79 - 2.62 (m, 3H), 2.62 - 2.44 (m, 2H), 2.44 - 2.24 (m, 4H), 2.16 - 2.10 (m, 1H), 1.37 (m, 6H). [M+H] m / z 458.488.

[1131] The following examples were prepared using similar methods as described above.

[1132]

[1133]

[1134]

[1135]

[1136] Example 548

[1137]

[1138] Step 1

[1139] Boc DAP OH (2 g, 9.79 mmol) was suspended in a mixture of THF (70.0 ml) and iPr2EtN (3.42 ml, 19.59 mmol) and then cooled to 0 °C. 4-Chlorobutyryl chloride (1.096 ml, 9.79 mmol) was then added dropwise (10:25 am). The reaction mixture was stirred at room temperature for approximately 40 minutes and then cooled to 0 °C. KOtBu (4.40 g, 39.2 mmol) was added in portions over a 2 minute period starting at 11:15. After 10 minutes, the reaction pH was adjusted to pH 1 with 1 N HCl and then extracted with ethyl acetate. The organic phase was washed with brine, dried over magnesium sulfate, filtered, and concentrated. The residue was chromatographed on silica gel eluting with 30-100% ethyl acetate in cyclohexane to yield (S)-2-((tert-butoxycarbonyl)amino)-3-(2-oxopyrrolidin-1-yl)propanoic acid (1.42 g, 2.87 mmol, 29% yield).1H NMR (400 MHz, DMSO-d6) δ 12.73 (s, 1H), 7.09 (d, J=8.5 Hz, 1H), 4.15 (td, J=8.5, 5.4 Hz, 1H), 3.59 (dd, J=13.7, 5.4 Hz, 1H), 3.40 - 3.28 (m, 4H), 2.24 - 2.11 (m, 2H), 1.91 - 1.80 (m, 2H), 1.37 (s, 9H). [M+Na] m / z 295.326.

[1140] Step 2

[1141] (S)-2-((tert-butoxycarbonyl)amino)-3-(2-oxopyrrolidin-1-yl)propanoic acid (389 mg, 1.43 mmol) was added to a solution of 4N HC1 in dioxane (4.3 mL, 12 eq. HC1) at room temperature. After 30 minutes at room temperature, the resulting white suspension was directly concentrated to yield (S)-2-amino-3-(2-oxopyrrolidin-1-yl)propanoic acid hydrochloride (408 mg, crude). The crude product was used without further purification as a white solid.1H NMR (400 MHz, Deuterium oxide) δ 4.17 (dd, J = 6.4, 4.2 Hz, 1H), 3.89 (dd, J = 15.1, 4.2 Hz, 1H), 3.79 (d, J = 6.4 Hz, 1H), 3.54 (td, J = 7.3, 1.6 Hz, 2H), 2.45 (dd, J = 9.1, 7.6 Hz, 2H), 2.08 (qd, J = 8.2, 6.9 Hz, 2H).

[1142] Step 3

[1143] (S)-2-amino-3-(2-oxopyrrolidin-1-yl)propanoic acid hydrochloride (20 mg, crude) was dissolved in a mixture of THF (0.48 mL) and water (0.48 mL) followed by the addition of solid NaHC03(40 mg, 0.48 mmol). The reaction mixture was cooled to 0 °C followed by the addition of CbzCl (15 uL, 0.105 mmol). The resulting mixture was stirred at room temperature for 3 days then partitioned between ethyl acetate and 1 N HC1. The organic phase was washed with brine, dried over magnesium sulfate, filtered, and concentrated to yield crude (S)-2-(((benzyloxy)carbonyl)amino)-3-(2-oxopyrrolidin-1-yl)propanoic acid which was used without further purification (assumed 100% yield). [M+H] m / z 307.322.

[1144] Step 4

[1145] HATU (35 mg, 0.092 mmol) and iPr2NEt (74 uL, 0.424 mmol) were added sequentially to a stirred solution of Compound 1-4 (29.5 mg, 0.11 mmol) and (S)-2-(((benzyloxy)carbonyl)amino)-3-(2-oxopyrrolidin-1-yl)propanoic acid (crude, assumed 0.085 mmol) in DMF (0.42 mL) at -10 °C. The mixture was allowed to warm to room temperature. The reaction was partitioned between ethyl acetate and water, then the organic layer was washed sequentially with saturated aqueous NaHC03and brine, dried over magnesium sulfate, filtered, and concentrated. The crude product residue (assumed 100% yield) was used directly in the next step without further purification. [M+H] m / z 520.468.

[1146] Step 5

[1147] Burgess reagent (122 mg, 0.51 mmol) was added to a solution of benzyl ((S)-1-((3R,5'S)-5'-carbamoyl-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-1-oxo-3-(2- oxopyrrolidin-1-yl)propan-2-yl)carbamate (assumed 0.085 mmol) at room temperature. After 45 min, MeOH was added and the reaction mixture was then treated with preparative HPLC for purification to yield benzyl ((S)-1-((3R,5'S)-5'-cyano-2-oxospiro[indoline-3,3'-pyrrolidin]-1'-yl)-1-oxo-3-(2- oxopyrrolidin-1-yl)propan-2-yl)carbamate (1.28 mg).1H NMR (400 MHz, Acetone-d6) δ 9.69 (s, 1H), 7.37 - 7.29 (m, 5H), 7.26 (td, J = 7.7, 1.2 Hz, 1H), 7.18 (d, J = 7.4 Hz, 1H), 7.00 (t, J = 7.8 Hz, 2H), 6.82 (d, J = 8.1 Hz, 1H), 5.16 (dd, J = 8.7, 7.0 Hz, 1H), 5.03 (d, J = 12.6 Hz, 1H), 4.95 (d, J = 12.6 Hz, 1H), 4.77 (td, J = 7.9, 4.9 Hz, 1H), 4.16 (q, J = 10.6 Hz, 2H), 3.68 (dd, J = 14.1, 7.8 Hz, 1H), 3.60 - 3.40 (m, 4H), 2.78 (dd, J = 13.4, 8.8 Hz, 1H), 2.66 (dd, J = 13.3, 7.1 Hz, 1H), 2.20 (td, J = 8.2, 2.1 Hz, 2H), 2.09 - 2.00 (m, 4H), 2.00 - 1.87 (m, 2H). [M+H] m / z 502.481.

[1148] Example 549

[1149]

[1150]

[1151] Step 1

[1152] A 500-mL round bottom flask was charged with (S)-2-amino-3-(lH-pyrrolo[2,3- b]pyridin-3-yl)propanoic acid (3 g, 14.62 mmol) and a magnetic stir bar. Methanol (50 mL) was added to produce a white suspension. The flask was flushed with nitrogen, sealed with a rubber septum, and cooled to 0 °C. Thionyl chloride (3.20 ml, 43.9 mmol) was added slowly under a positive pressure of nitrogen, then the flask was spun into an ice bath. The resulting light yellow solution was stirred for 10 minutes then allowed to warm to near room temperature, then heated to reflux for 2 hours. The reaction solution was evaporated to produce a white solid which was carried forward to the next step without further purification.

[1153] Step 2

[1154] A 500-mL round bottom flask containing crude product 7-azasemino acid methyl ester was charged with a magnetic stir bar, then pyridine (37 mL) was added, forming a white gas and noting a slight exotherm. The reaction mixture was sonicated until a light gold solution was produced. Formaldehyde (1.197 ml, 37% w / w in water containing 10% methanol, 16.08 mmol, 1.10 eq) was added. A condenser was attached to the reaction vessel and heated to reflux and stirred for 1 hour, while the reaction was judged complete by LCMS. The reaction vessel was protected in an ice bath for 20 minutes, producing a thick off-white slurry. The slurry was then filtered through a sintered funnel and the filter cake was washed with pre-chilled pyridine (2 x 20 mL), then DCM (10 mL). The resulting solid was dried under a stream of nitrogen overnight, producing β-carboline 549-1 white solid which was used directly in the next step without further purification.

[1155] Step 3

[1156] A 250-mL round bottom flask containing crude product β-carboline 549-1 (3.38 g, 14.62 mmol) and a magnetic stir bar was charged with water (7.5 mL) and THF (30 mL) to produce a hazy solution. The reaction vessel was cooled to 0 °C and triethylamine (6.11 ml, 43.9 mmol) was added, followed by di-tert-butyl dicarbonate (8.49 ml, 36.6 mmol). The reaction vessel was flushed with nitrogen and placed under a positive pressure of nitrogen. After stirring for 2 days, the reaction mixture was partitioned between ethyl acetate and water and the layers were separated. The aqueous phase was then extracted with ethyl acetate (3 x 20 mL), the combined organic phases were dried over sodium sulfate, filtered, and concentrated to produce a yellow residue. The residue was purified by flash chromatography on a silica gel column eluted with ethyl acetate in cyclohexane to produce colorless residue 549-2 (1.61 g, 33% overall yield over three steps). 1H NMR (400 MHz, DMSO-d6), mixture of rotamers: δ 11.40 (app. d, J = 24.6 Hz, 1H), 8.13 (dd, J = 4.8, 1.5 Hz, 1H), 7.85 (dd, J = 7.8, 1.6 Hz, 1H), 7.02 (dd, J = 7.8, 4.7 Hz, 1H), 5.31 - 5.13 (m, 1H), 4.75 (t, J = 16.9 Hz, 1H), 4.49 - 4.27 (m, 1H), 3.57 (d, J = 8.0 Hz, 3H), 3.27 (m, 1H), 3.00 (dd, J = 15.5, 7.1 Hz, 1H), 1.46 (app. d, J = 17.6 Hz, 9H).

[1157] Step 4

[1158] Compound 549-2 (1.60 g, 4.83 mmol) was added and dissolved in THF (22.80 mL) into a 1000 mL round bottom flask containing a magnetic stir bar. Water (2.85 mL) was added to give a colorless, magnetically stirred solution which was cooled to 0 °C. N-Bromosuccinimide (0.859 g, 4.83 mmol) was added in portions over several minutes to give a yellow solution. The flask was then capped with a rubber septum and glacial acetic acid (1.935 mL, 33.8 mmol) was added. The color of the reaction mixture solution darkened significantly to a thick orange-yellow color and stirring was continued for 30 minutes. Potassium carbonate (3.34 g, 24.14 mmol) and water were then added and the reaction mixture was allowed to warm to room temperature. The layers were separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, poured off, and concentrated under reduced pressure to give a tan gum or foam. Purification by flash chromatography on silica gel eluting with ethyl acetate in cyclohexane gave the pure product 549-3 (220 mgs) as a single diastereomer and impurity product (1.44 g, d.r. ~ 3: 1 desired product / non-desired product) as a mixture of diastereomers.

[1159] Step 5

[1160] To a 100-mL round bottom flask containing 549-3 (220 mg, 0.633 mmol) and a magnetic stir bar was added methanamine (4.52 mL of a 7M solution, 31.7 mmol). The flask was sealed with a plastic film with a needle and the yellow reaction solution was heated to 55 °C and stirred for 6 days. The reaction mixture was then evaporated until a tan solid 549-4 remained.

[1161] Step 6

[1162] The crude product amine 549-4 was suspended in a solution of HCI in dioxane (3 mL of a 4.0 M solution, 12 mmol). The resulting suspension was stirred vigorously at room temperature overnight, then evaporated to yield the hydrochloride salt of amine 549-5 as an off-white solid (95 mg, 49% yield over three steps). 1 H NMR (400 MHz, Deuterium oxide) δ 8.19 (dd, J = 5.7, 1.4 Hz, 1H), 8.01 (ddd, J = 7.5, 1.5, 0.6 Hz, 1H), 7.30 (dd, J = 7.7, 5.9 Hz, 1H), 4.94 (t, J = 8.5 Hz, 1H), 3.94 (t, J = 13.2 Hz, 1H), 3.81 (d, J = 12.9 Hz, 1H), 2.92 (dd, J = 14.1, 9.1 Hz, 1H), 2.73 (dd, J = 14.1, 7.9 Hz, 1H).

[1163] Step 7

[1164] To a 40 mL glass reaction vessel was added compound 549-5 (181 mg, 0.593 mmol), N-(tert-butoxycarbonyl)-N-methyl-L-leucine (198 mg, 0.807 mmol), and a magnetic stir bar. DCM (3 mL) and DMF (0.750 mL) were added, resulting in a white suspension. The reaction mixture was cooled to 0 °C; N-methylmorpholine (0.261 mL, 2.373 mmol) was added, followed by HATU (226 mg, 0.593 mmol). The reaction vessel was flushed with nitrogen and the reaction mixture was allowed to warm to room temperature with stirring overnight. The reaction mixture was then evaporated on celite and the crude mixture was purified using flash chromatography on silica gel (4 g) eluted with DCM / MeOH to yield the intermediate pure product, which was then purified by reverse phase HPLC to yield the product 549-6 as a colorless solid.

[1165] Step 8

[1166] The 549-6 obtained from the above step was poured into a clear glass vessel containing a magnetic stir bar and treated with a solution of HCI in dioxane (2 mL of a 4.0 M solution, 8.00 mmol). The resulting homogeneous suspension was stirred for 11 hours, the volatile material was removed under reduced pressure, and white solid 549-7 was produced.

[1167] Step 9

[1168] Compound 549-7 obtained from the above procedure was dissolved in DCM (0.8 mL) and DMF (0.200 mL) to give a white suspension. N-methylmorpholine (0.0717 mL, 0.652 mmol) was added; the resulting mixture immediately became clear to give a colorless liquid. HATU (62.0 mg, 0.163 mmol) was added under a stream of nitrogen, followed by 4,6-difluoro-lH-indole-2-carboxylic acid (32.1 mg, 0.163 mmol). The reaction vessel was flushed with nitrogen, capped, and the resulting reaction mixture was stirred for 3 h at which time the reaction was quenched by partitioning into one-quarter saturated aqueous sodium bicarbonate solution and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate (3 x 10 mL); the combined organic was dried over sodium sulfate, filtered, and concentrated under reduced pressure to give an off-white residue 549-8.

[1169] Step 10

[1170] To a 40-mL glass reaction vessel containing crude amine compound 549-8 and a magnetic stir bar was added DCM (1.25 mL). The reaction vessel was flushed with nitrogen, cooled to 0 °C, and triethylamine (0.1555 mL, 1.116 mmol) was poured in. Trifluoroacetic anhydride (0.0787 mL, 0.557 mmol) was then added dropwise to give a yellow reaction mixture, the reaction mixture was allowed to warm to room temperature, and stirred for 1.25 h at which time complete consumption of starting material was indicated by LCMS. The reaction was quenched with saturated aqueous sodium bicarbonate solution, the layers were separated, the organic was washed with brine, and concentrated under reduced pressure to give a yellow gum. The crude product was purified by reverse phase HPLC to give a colorless film which was lyophilized to give example 549 as a free-flowing white powder (34 mgs, 11% yield over four steps). 1H NMR (400 MHz, Acetone-d6) δ 10.87 (s, 1H), 10.26 (s, 1H), 7.93 (s, 1H), 7.36 (d, J = 7.3 Hz, 1H), 7.12 - 7.00 (m, 2H), 6.81 - 6.67 (m, 2H), 5.55 (dd, J = 9.7, 5.4 Hz, 1H), 5.21 (t, J = 8.3 Hz, 1H), 4.43 (d, J = 10.8 Hz, 1H), 4.03 (d, J = 10.8 Hz, 1H), 3.47 (s, 3H), 2.83 (ddd, J = 13.4, 8.6, 1.2 Hz, 1H), 2.72 (dd, J = 13.4, 8.0 Hz, 1H), 1.96 (ddd, J = 14.4, 9.7, 4.9 Hz, 1H), 1.76 (ddd, J = 14.2, 8.8, 5.4 Hz, 1H), 1.62 (dtd, J = 8.9, 6.6, 4.9 Hz, 1H), 1.00 (d, J = 6.7 Hz, 3H), 0.94 (d, J = 6.5 Hz, 3H). LCMS [M+H] + 521.0.

[1171] The following examples were prepared using similar methods as described above:

[1172]

[1173]

[1174]

[1175] Biological Activity

[1176] SARS-CoV-2 3C-like (3CL) protease fluorescence assay (FRET): Recombinant SARS-CoV-2 3CL-protease was expressed and purified. TAMRA-SITSAVLQSGFRKMK-Dabcyl-OH peptide 3CLpro substrate was synthesized. Black, low volume, round bottom 385-well microplates were used. In a routine experiment, 0.85 pL of the compound to be tested was dissolved in DMSO and then incubated with SARS-CoV-2 3CL-protease (10 nM) in 10 pL assay buffer (50 mM HEPES [pH 7.5], 1 mM DTT, 0.01% BSA, 0.01% Triton-X 100) for 30 min at room temperature. Subsequently, 10 pL of 3CL-protease substrate (40 mM) in assay buffer was added and the experiment was continuously monitored for 1 h in an Envision Multimode Plate Reader working in fluorescence kinetics mode with an excitation wavelength of 540 nm and an emission wavelength of 580 nm at room temperature. No compound (only DMSO) and no enzyme controls were included in each microplate and run routinely according to this method. All experiments were run in duplicate.

[1177] Data analysis: SARS-CoV-2 3CL-protease activity was measured as the initial velocity of the linear phase (RFU / s) and this value was used to determine the percentage of residual activity of the test compounds (0-10 pM) in relation to the reference samples DMSO (100% activity) and no enzyme (0% activity). Data were fitted in GraphPad Prism 7 to determine IC 50 All experiments were run in duplicate and IC 50 Range readings are shown as follows: A < 0.1 mM; B 0.1-1 mM; C > 1 mM.

[1178] Table 1. Activity summary

[1179]

[1180]

[1181]

[1182]

[1183]

[1184]

[1185]

[1186]

[1187] 229E Experimental Protocol

[1188] Virus Stock Preparation MRC-5 cells (a diploid cell line composed of fibroblasts, originally developed from lung tissue of a 14-week-old aborted Caucasian male fetus) were used for culture of 229E human coronavirus (hCoV). Flasks were inoculated with hCoV-229E and when cytopathic effect (CPE) was greater than 70%, virus stocks were collected. Virus stocks in growth media (EMEM, 1% Penn / Strep, 1% non-essential amino acids, 10% heat inactivated FBS) plus 5% glycerol were snap frozen using liquid nitrogen and stored at -80°C. Virus stocks were quantified by TCID50 (50% tissue culture infective dose median) assay as described elsewhere. 50 Virus stock titers were quantified by TCID50 (50% tissue culture infective dose median) assay.

[1189] 229E Live Virus Experiments:The assay uses 384-well black cell culture treated plastic clear bottom plates. Using an ECHO liquid dispenser, a 3-fold serial dilution of the reference and test compounds suspended in dimethyl sulfoxide are added to the plate wells in duplicate, with a total volume of 125 nL / well. MRC-5 cells at passage 17 or below are seeded into the inner 240 wells of the 384-well plate at 1500 cells per well in a volume of 12.5 μL using growth media. Virus stock is then added to each well at a multiplicity of infection (MOI) of 0.05 in a volume of 12.5 μL per well, bringing the total volume in each well to approximately 25 μL. Each plate has a row of 20 wells containing cells plus DMSO and virus but no compound (positive control, maximum CPE, minimum ATPlite signal), a row of wells containing cells plus dimethyl sulfoxide but no compound or virus (negative control, minimum CPE, maximum ATPlite signal), and a row of wells containing no cells or virus or compound (background plate / reagent control). To the control wells containing cells but no virus, an additional 12.5 μL of growth media is added, containing the same amount of glycerol as the wells receiving virus stock, to maintain equal amounts of media and volume conditions. The outer 2 rows / columns of wells are filled with 30 μL moat media (DMEM, 1% Penn / Strep) as a heat and evaporation barrier for the test wells. After all components are added, the plate is tapped gently on the side to encourage even distribution of the cells throughout the well. After confirming cell distribution, the plate is incubated at 34°C in a CO2humidified incubator for 6 days. After the 6-day incubation period, the plates are read using ATPlite (12.5 μL per well) which quantifies the amount of ATP (a measure of cell health) present in each well. The plate is read using an Envision luminometer. These data are used to calculate the percent cell health of each well relative to the negative control wells, and the EC50 50 are calculated and analyzed using four parameter logistic curve fit analysis using ExcelFit software.

[1190] All experiments are run in duplicate and EC 50 ranges are as follows: A < 0.1 μM; B 0.1-1 μM; C > 1 μM.

[1191] Table 2. Activity Summary

[1192]

[1193]

[1194]

[1195]

[1196]

[1197]

[1198]

[1199]

[1200] While the application has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the application, the scope of the application being measured by the appended claims.

Claims

1. A compound of formula (Ia-2) as depicted in the following scheme: or a pharmaceutically acceptable salt thereof, wherein: A is selected from -R 11 , -OR 12 , and -NR 13 R 14 ; B is pyridyl or optionally substituted phenyl; X is selected from: 1) -CN; 2) -C(O)R 15 ; 3) -CH(OH)SO3R 16 ; and 4) -C(O)C(O)NR 13 R 14 ; R1is optionally substituted -C1-C6alkyl, optionally substituted -C3-C6cycloalkyl, optionally substituted C3-C6cycloalkyl-C1-C2-alkyl-, optionally substituted aryl, optionally substituted aryl-C1-C6-alkyl or optionally substituted heteroaryl-C1-C6-alkyl, R3is selected from hydrogen, -Me, -Et, -Pr, -i-Pr, -propenyl, -CF3, -CD3and cyclopropyl; R4is selected from hydrogen, -Me, -Et, -Pr, -i-Pr, -propenyl, -CF3and cyclopropyl; R 11 and R 12 are independently of one another selected from the group consisting of 1) optionally substituted -C1-C8alkyl; 2) optionally substituted -C2-C8alkenyl; 3) optionally substituted -C2-C8alkynyl; 4) optionally substituted -C3-C8cycloalkyl; 5) optionally substituted 3- to 8-membered heterocycloalkyl; 6) optionally substituted aryl; 7) optionally substituted aryl-C1-C6-alkyl; 8) optionally substituted heteroaryl; and 9) optionally substituted heteroaryl-C1-C6-alkyl; R 13 and R 14 are each independently selected from the group consisting of: 1) hydrogen; 2) optionally substituted -C1-C8alkyl; 3) optionally substituted -C2-C8alkenyl; 4) optionally substituted -C2-C8alkynyl; 5) optionally substituted -C3-C8cycloalkyl; 6) optionally substituted 3- to 8-membered heterocycloalkyl; 7) optionally substituted aryl; 8) optionally substituted aryl-C1-C6-alkyl; 9) optionally substituted heteroaryl; and 10) optionally substituted heteroaryl-C1-C6-alkyl; R 15 is hydrogen, hydroxyl, or optionally substituted -Ci-C8alkyl; R 16 is hydrogen or Na + ; wherein: (i) each optionally substituted substituent is substituted with 1 to 3 substituents selected from halogen, C1-C4-alkyl, halo-C1-C4-alkyl, C2-C4-alkenyl, halo-C2-C4-alkenyl, C3-C6-cycloalkyl, C1-C4-alkoxy, halo-C1-C4-alkoxy, acyl, -CN, -OH, NH2, C1-C4-alkylamino, di(C1-C4-alkyl)amino and NO2; (ii) each aryl is phenyl or naphthyl; (iii) each heteroaryl is a monocyclic or bicyclic aromatic radical having one or more ring atoms selected from S, O and N; wherein the remaining ring atoms are carbon; and (iv) each heterocyclyl is a non-aromatic ring or a fused, spiro or bridged system of bicyclic or tricyclic radicals wherein i) each ring contains at least 1 heteroatom independently selected from oxygen, sulfur and nitrogen, ii) each ring system can be saturated or unsaturated, iii) said nitrogen and sulfur heteroatoms can be optionally oxidized, iv) said nitrogen heteroatoms can be optionally quaternized, v) any of the above rings can be fused to a benzene ring, and vi) the remaining ring atoms are carbon atoms which can be optionally substituted with oxygen or optionally substituted with an exogenous ring alkene double bond.

2. The compound of claim 1, wherein, A is selected from any of the following groups and is optionally substituted:

3. The compound of claim 1 or 2, wherein, X is -CN.

4. The compound according to claim 1 represented by one of the following formulae (VIII-1a) to (VIII-5a), or a pharmaceutically acceptable salt thereof: wherein R9is halogen; A, R1and R3are defined as in claim 1.

5. The compound according to claim 1 represented by one of the following formulae (XIII-1) to (XIII-6), or a pharmaceutically acceptable salt thereof: wherein n is 0, 1, 2, 3, or 4; m is 0, 1, 2, 3, 4 or 5; v is 0, 1 or 2; each R9is halogen; each R9' is independently selected from the group consisting of: halogen; -CN; -OR 11 each R 11 is hydrogen, C1-C4-alkyl or halo-C1-C4-alkyl; -NR 13 R 14 wherein R 13 and R 14 are independently of each other selected from hydrogen and -Ci-C4alkyl; halo -C1-C4-alkyl and optionally substituted -C3-C8-cycloalkyl; R 10 is optionally substituted -C1-C4alkyl or optionally substituted -C3-C6cycloalkyl; R4is as defined in claim 1.

6. The compound according to claim 1 represented by formula (XI-3): wherein, each n is 0, 1, 2, 3 or 4; v is 0; R3is hydrogen, methyl or CD3; R1is C1-C6-alkyl or aryl-C1-C6-alkyl; R4is hydrogen; each R9’ is halogen or C1-C4-alkoxy; and each R9is halogen.

7. The compound according to claim 6 represented by the following formula 8. The compound according to claim 7 represented by the following formula wherein R3is hydrogen or methyl.

9. The compound according to claim 1 represented by the following formula wherein, X is CN; R3is hydrogen, methyl or CD3; R1is C1-C6-alkyl, aryl-C1-C6-alkyl or optionally substituted C3-C6-cycloalkyl-C1-C2-alkyl; R4is hydrogen; and R9is C1-C6-alkyl or phenyl optionally substituted by one to three substituents independently selected from halogen, C1-C6-alkyl and C1-C6-haloalkyl.

10. The compound according to claim 1 represented by one of the following formulae (IX-1a) to (IX-5a), wherein: R9is halogen; R3is hydrogen or methyl; A is selected from the following groups: and R1is selected from the following groups:

11. A compound selected from the following compounds represented by the following formulae or a pharmaceutically acceptable salt thereof:

12. The compound according to claim 11, wherein the compound is 13. The compound according to claim 11, wherein the compound is 14. The compound according to claim 11, wherein the compound is 15. The compound according to claim 11, wherein the compound is 16. The compound according to claim 11, wherein the compound is 17. The compound according to claim 11, wherein the compound is 18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17, and a pharmaceutically acceptable carrier or excipient.

19. Use of a compound according to any one of claims 1 to 17 for the manufacture of a medicament for the treatment or prophylaxis of a coronavirus infection.

20. The use according to claim 19, wherein, The coronavirus is 229E, NL63, OC43, HKU1, SARS-CoV or MERS coronavirus.

21. The use according to claim 20, wherein the coronavirus is SARS-CoV-2.

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