NBD1 modulators and methods of using the same

Abstract

The present disclosure includes, among other things, CFTR modulators, pharmaceutical compositions, and methods of making and using the same.

Description

[0001] Attorney Docket No. SLI-021WO

[0002] NBD1 MODULATORS AND METHODS OF USING THE SAME Background

[0003]

[0001] Cystic fibrosis (CF), an autosomal recessive disorder, is caused by functional deficiency of the cAMP-activated plasma membrane chloride channel, cystic fibrosis transmembrane conductance regulator (CFTR), which results in pulmonary and other complications. The gene encoding CFTR has been identified and sequenced (See Gregory, R. J. et al. (1990) Nature 347:382-386; Rich, D. P. et al. (1990) Nature 347:358-362); Riordan, J. R. et al. (1989) Science 245: 1066-1073). CFTR, a member of the ATP binding cassette (ABC) superfamily is composed of two six membrane-spanning domains (MSD1 and MSD2), two nucleotide bind domains (NBD1 and NBD2), a regulatory region (R) and four cytosolic loops (ICL4). CFTR protein is located primarily in the apical membrane of epithelial cells where it functions to conduct anions, including chloride, bicarbonate, and thiocyanate into and out of the cell. CFTR may have a regulatory role over other electrolyte channels, including the epithelial sodium channel ENaC.

[0004]

[0002] In cystic fibrosis patients, the absence or dysfunction of CFTR leads to exocrine gland dysfunction and a multisystem disease, characterized by pancreatic insufficiency and malabsorption, as well as abnormal mucociliary clearance in the lung, mucostasis, chronic lung infection and inflammation, decreased lung function and ultimately respiratory failure.

[0005]

[0003] While more than 1,900 mutations have been identified in the CFTR gene, a detailed understanding of how each CFTR mutation may impact channel function is known for only a few. (Derichs, European Respiratory Review. 22:127, 58-65 (2013)). The most frequent CFTR mutation is the in-frame deletion of phenylalanine at residue 508 (AF508) in the first nucleotide binding domain (NBD1). Over 70% of cystic fibrosis patients have a deletion at residue 508 in at least one CFTR allele. The loss of this key phenylalanine renders NBD1 conformationally unstable at physiological temperature and compromises the integrity of the interdomain interface between NBD 1 and CFTR’ s second transmembrane domain (ICL4). The AF508 mutation causes production of misfolded CFTR protein which, rather than traffic to the plasma membrane, is instead retained in the endoplasmic reticulum and targeted for degradation by the ubiquitin-proteasome system.

[0006]

[0004] The loss of a functional CFTR channel at the plasma membrane disrupts ionic homeostasis and airway surface hydration leading to reduced lung function. Reduced periciliary liquid volume and increased mucus viscosity impede mucociliary clearance resulting in chronic infection and inflammation. In the lung, the loss of CFTR-function leads

[0007] 1

[0008] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0009] to numerous physiological effects downstream of altered anion conductance that result in the dysfunction of additional organs such as the pancreas, intestine, and gall bladder.

[0010]

[0005] By studying the mechanistic aspects of CFTR misfolding and corrections, small molecules have been identified as CF modulators, which can function as stabilizers.

[0011]

[0006] Despite the identification of compounds that modulate CFTR, there is no cure for this fatal disease and identification of new compounds and new methods of therapy are needed as well as new methods for treating or lessening the severity of cystic fibrosis and other CFTR mediated conditions and diseases in a patient.

[0012] Summary

[0013]

[0007] The present disclosure includes a compound of formula I:

[0014]

[0015] I

[0016] or a pharmaceutically acceptable salt thereof. Additionally, the present disclosure includes, among other things, pharmaceutical compositions, methods of using and methods of making a compound of formula I.

[0017] 2

[0018] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0019] Detailed Description

[0020]

[0008] In some embodiments, the present disclosure includes a compound of Formula I:

[0021]

[0022] I

[0023] or a pharmaceutically acceptable salt thereof,

[0024] wherein

[0025] L1is an optionally substituted Ci-6 alkylene chain wherein 1-3 of the methylene units is optionally and independently replaced by -O-, -N(R2)-, -C(O)-, -S-, -S(O)-, an<o>

[0026] optionally substituted 3-6 membered carbocyclyl,, or optionally substituted C2 alkenylene;

[0027] L2is an optionally substituted C1-6 alkylene chain wherein 1-3 of the methylene units is optionally and independently replaced by -C(CDs)2-, -O-, -N(R2), -C(O)-, -S-, -S(O)-, an optionally substituted 3-6 membered carbocyclyl, or optionally substituted C2 alkenylene;

[0028] Ring A is an optionally substituted 5-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S;

[0029] Ring C is optionally substituted phenyl or optionally substituted 5-6-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S;

[0030] X is selected from the group consisting of -O-, -S-, -CH2-, -C(OH)H-, -S(O)-, -C(O)-, - S(O)2-, -CFH-, -CF2-, and -N(R2)-;

[0031] 3

[0032] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0033] each RAis independently selected from the group consisting of halogen, cyano, optionally substituted Ci-Ce aliphatic, optionally substituted Ci-Ce alkoxy, and -CD3;

[0034] each RBis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci-Ce aliphatic and optionally substituted Ci-Ce alkoxy;

[0035] each Rcis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci- aliphatic; wherein at least one instance of Rcis -OR2or -N(R2)2;

[0036] each RDis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, optionally substituted Ci-Ce aliphatic, optionally substituted C1-C3 alkoxy, optionally substituted 5-6-membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S, wherein each RDis optionally substituted with 1-6 instances of Rd;

[0037] wherein two instances of RDmay be taken together to form an optionally substituted 5-7 membered carbocyclic ring, optionally substituted 5-6- membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S; each Rdis independently selected from the group consisting of hydrogen, -OH, -CD3, - C(O)N(R2)2, C(O)OR2, -OR2, -N(R2)2, -S(O)2R2, optionally substituted 5-6- membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S;

[0038] R1is selected from the group consisting of hydrogen, cyano, -OR2, -(CH2)o-3N(R2)2, optionally substituted C1-C3 aliphatic, and -CD3;

[0039] each R2is independently selected from hydrogen, optionally substituted Ci-Ce aliphatic, - OH, Ci-Ce alkoxy, -S(O)2(optionally substituted Ci-Ce aliphatic);

[0040] n is 0, 1, 2, or 3;

[0041] p is 0, 1, 2, 3, or 4;

[0042] q is 1, or 2; and

[0043] 4

[0044] IPTS / 200162816.3 Attorney Docket No. SLI-021WO r is 0, 1, 2, 3, 4, or 5.

[0045]

[0009] In some embodiments, a compound of the present disclosure is of Formula (I-a), (I-b), (I-c), (I-d), (I-e), or (I-f):

[0046] (I-a) (I-b)

[0047]

[0048] 5

[0049] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0050]

[0051] (I-e) (I-f)

[0052] or a pharmaceutically acceptable salt thereof.

[0053] X

[0054]

[0010] In some embodiments, X is selected from the group consisting of -O-, -S-, -CH2-, -C(OH)H-, -S(O)-, -C(O)-, -S(O)2-, -CFH-, -CF2-, and -N(R2)-. In some embodiments, X is -O-. In some embodiments, X is -S-. In some embodiments, X is -C(OH)H-. In some embodiments, X is -S(O)-. In some embodiments, X is -S(O)2-. In some embodiments, X is -CH2-. In some embodiments, X is -C(O)-.

[0055] Ring A

[0056] [Oil] In some embodiments, Ring A is an optionally substituted 5-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S. In some embodiments, Ring A is an optionally substituted 5-membered heteroaryl comprising 1-3 heteroatoms each independently selected from the group consisting of N, O, and S. In some embodiments, Ring A is an optionally substituted 5-membered heteroaryl comprising 1-4 nitrogen atoms. In some embodiments, Ring A is an optionally substituted 5-membered heteroaryl comprising 2 nitrogen atoms. In some embodiments, Ring A is an optionally substituted 5-membered heteroaryl comprising 3 nitrogen atoms. In some embodiments, Ring A is optionally substituted triazolyl or optionally substituted imidazolyl. In some embodiments, Ring A is optionally substituted 1,3,4-triazolyl or optionally substituted imidazolyl.

[0057]

[0012] In some embodiments, Ring A is optionally substituted 1,3,4-triazolyl. In some

[0058] 6

[0059] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0060] embodiments Ring A is

[0061]

[0062]

[0013] In some embodiments, Ring A is optionally substituted imidazolyl. In some

[0063] embodiments, Ring A is Ring A is

[0064]

[0065] Ring C

[0066]

[0014] In some embodiments, Ring C is optionally substituted phenyl or optionally substituted 5-6-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S.

[0067]

[0015] In some embodiments, Ring C is optionally substituted phenyl. In some embodiments,

[0068] Ring C

[0069]

[0070] is

[0071]

[0016] In some embodiments, Ring C is selected from the group consisting of

[0072]

[0073]

[0017] In some embodiments, Ring C is selected from the group consisting of

[0074]

[0075]

[0018] In some embodiments, Ring C is optionally substituted 5-6-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S. In some embodiments, Ring C is optionally substituted 6-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S. In some embodiments, Ring C is optionally substituted pyridinyl.

[0076]

[0019] In some embodiments, Ring C is selected from the group consisting of

[0077] 7

[0078] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0079]

[0080]

[0020] In some embodiments, Ring C is

[0081]

[0082]

[0021] In some embodiments, Ring C is optionally substituted 5-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S. In some embodiments, Ring C is optionally substituted thiazolyl. In some embodiments,

[0083] NH2

[0084] In some embodiments, Ring C is

[0085] R

[0086]

[0087] A

[0088]

[0022] In some embodiments, each RAis independently selected from the group consisting of halogen, cyano, optionally substituted CI-C aliphatic. optionally substituted Ci-Ce alkoxy, and -CDs. In some embodiments, each RAis independently optionally substituted Ci-Ce aliphatic or -CDs. In some embodiments, each RAis -CDs. In some embodiments, each RAis independently optionally substituted Ci-Ce aliphatic. In some embodiments, each RAis independently optionally substituted Ci-Cs aliphatic. In some embodiments, each RAis independently optionally substituted C1-C2 aliphatic. In some embodiments, each RAis independently -CH3 or -CH2CH2N(H)CHs. In some embodiments a Ci-Ce aliphatic of RAis a Ci-Ce alkyl.

[0089] RB

[0090]

[0023] In some embodiments, each RBis independently selected from the group consisting of halogen, cyano, -C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci-Ce aliphatic and optionally substituted Ci-Ce alkoxy. In some embodiments, RBis halogen. In some embodiments, RBis fluoro. In some embodiments a C1-C6 aliphatic of RBis a Ci-Ce alkyl. Rc

[0091]

[0024] In some embodiments, each Rcis independently selected from the group consisting of halogen, cyano, -C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci-Ce

[0092] 8

[0093] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0094] aliphatic; wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments a Ci-Ce aliphatic of Rcis a Ci-Ce alkyl. In some embodiments Rcis independently selected from the group consisting of halogen, -OR2, -N(R2)2, and optionally substituted Ci-Ce aliphatic, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, an optionally substituted Ci-Ce aliphatic of Rcis an optionally substituted Ci-Ce alkyl. In some embodiments, each Rcis independently halogen, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, each Rcis independently selected from the group consisting of -F, -Cl, -I, -OR2, and -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -OR2or -N(R2)2. n some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -OR2. In some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -N(R2)2. In some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -OH or -NH2. In some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -OH. In some embodiments, at least one instance of Rcis -F and at least one instance of Rcis -NH2.

[0095]

[0025] In some embodiments, Rcis optionally substituted Ci-Ce aliphatic, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, at least one instance of Rcis optionally substituted Ci-Ce aliphatic and at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, an optionally substituted Ci-Ce aliphatic of Rcis an optionally substituted Ci-Ce alkyl. In some embodiments, Rcis C1-C3 alkynyl, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments, Rcis -C=CH, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2. In some embodiments a Ci-Ce aliphatic of Rcis a Ci-Ce alkynyl. In some embodiments, Rcis a C1-C2 alkynyl. In some embodiments, at least one instance of Rcis -C=CH, -OR2or -N(R2)2, and at least one instance of Rcis -OR2or -N(R2)2.

[0096] RD

[0097]

[0026] In some embodiments, each RDis independently selected from the group consisting of halogen, cyano, -C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, optionally substituted Ci-Ce aliphatic, optionally substituted C1-C3 alkoxy, optionally substituted 5-6-membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S.

[0098]

[0027] In some embodiments, RDis optionally substituted with 1 -6 instances of Rd.

[0099]

[0028] In some embodiments, two instances of RDmay be taken together to form an optionally substituted 5-7 membered carbocyclic ring, optionally substituted 5-6-membered heteroaryl,

[0100] 9

[0101] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0102] or optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, two instances of RDmay be taken together to form an optionally substituted carbocyclic ring. In some embodiments, two instances of RDmay be taken together to form an optionally substituted 5-6-membered heteroaryl. In some embodiments, two instances of RDmay be taken together to form an optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S.

[0103]

[0029] In some embodiments, a Ci-Ce aliphatic of RDis a Ci-Ce alkyl. In some embodiments, each RDis independently selected from the group consisting of halogen and Ci-Ce aliphatic. In some embodiments, a Ci-Ce aliphatic of RDis a Ci-Ce alkyl. In some embodiments, RDis independently halogen. In some embodiments, RDis -F. In some embodiments, each RDis optionally substituted Ci -C6 aliphatic. In some embodiments, a Ci-Ce aliphatic of RDis a Ci-Ce alkyl. In some embodiments, RDis optionally substituted C1-C4 aliphatic. In some embodiments, a C1-C4 aliphatic of Rcis a C1-C4 alkyl.

[0104]

[0030] In some embodiments, at least one instance of RDis independently selected from the

[0105] list consisting of

[0106]

[0107]

[0031] In some embodiments, Rdis independently selected from the group consisting of hydrogen, -OH, -CD3, -C(O)N(R2)2, C(O)OR2, -OR2, -N(R2)2, -S(O)2R2, optionally substituted 5-6-membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, Rdis independently selected from the group consisting of hydrogen, -OH, -CD3, and -C(O)OR2. In some embodiments, Rdis independently selected from -OH and -C(O)OR2. In some embodiments, Rdis -OH. In some embodiments, Rdis -C(O)OR2. In some embodiments, the R2of -C(O)OR2is hydrogen. In some embodiments, Rdis -C(O)OH. Attorney Docket No. SLI-021WO

[0108] R’

[0109]

[0032] In some embodiments, R1is selected from the group consisting of hydrogen, cyano, -OR2, -(CH2)O-SN(R2)2, optionally substituted C1-C3 aliphatic, and -CD3. In some embodiments, a C1-C3 aliphatic is a C1-C3 alkyl. In some embodiments, R1is hydrogen. In some embodiments, R1is cyano. In some embodiments, R1is OR2. In some embodiments, R1is -(CH2)O-3N(R2)2. In some embodiments, R1is -N(R2)2. In some embodiments, R1is -(CH2)N(R2)2. In some embodiments, R1is -(CH2)2N(R2)2. In some embodiments, R1is -(CH2)3N(R2)2. In some embodiments, R1is C1-C3 aliphatic. In some embodiments, a C1-C3 aliphatic is a C1-C3 alkyl. In some embodiments, R1is CD3.

[0110] R2

[0111]

[0033] In some embodiments, each R2is independently selected from hydrogen, optionally substituted C1-C6 aliphatic, -OH, C1-C6 alkoxy, -S(O)2(optionally substituted Ci-Ce aliphatic). In some embodiments a Ci-Ce aliphatic of R2is a Ci-Ce alkyl. In some embodiments, R2is independently hydrogen. In some embodiments, R2is independently optionally substituted Ci-Ce aliphatic. In some embodiments, R2is independently -OH. In some embodiments, R2is independently Ci-Ce alkoxy. In some embodiments, R2is independently -S(O)2(optionally substituted Ci-Ce aliphatic). In some embodiments, an optionally substituted Ci-Ce aliphatic of R2is an optionally substituted Ci-Ce alkyl.

[0112]

[0034] In some embodiments, the present disclosure includes compounds listed in Table 1.

[0113]

[0114] 11

[0115] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0116]

[0117] 12

[0118] IPTS / 200162816.3 Attorney Docket No. SLI-021WO 9

[0119] II "} NH2

[0120] A

[0121] /

[0122] F M C

[0123] X

[0124] 10 z

[0125] ZD\^

[0126] \ / — o

[0127] > Q

[0128] M- 11 F

[0129] F X-^VN-

[0130] H^S

[0131] O"'VA

[0132] 12 o. P V

[0133] ci (S.

[0134] JI A I

[0135] ci>

[0136] HO fV 'N

[0137] F

[0138] Diastereomer 1

[0139] 13

[0140] F

[0141] Diastereomer 2

[0142] 14 o.9 V

[0143] c,,

[0144] ^NyU V! A^A^A, OH

[0145] F

[0146] 15

[0147] 0 0-X M / N

[0148] N'-’WNj 0

[0149] H, N-M

[0150]

[0151] 13

[0152] IPTS / 200162816.3 Attorney Docket No. SLI-021WO 16 F HN—

[0153] F jCXX- N

[0154] FYVNJ / N

[0155] XX

[0156] D °z0 " / A\

[0157] 17 F HN—

[0158] FXFA N

[0159] y / °n"

[0160] f

[0161] A °'A A

[0162] o A

[0163] 18

[0164] 19

[0165] 0 0A N / N

[0166] H2ANA

[0167] O'TT A

[0168] 20

[0169] r, 0^ N^ / N

[0170] 21

[0171] .FA A If ANH A2

[0172] ;N^ F0Z

[0173] 22

[0174]

[0175] 14

[0176] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0177]

[0178] 15

[0179] IPTS / 200162816.3 Attorney Docket No. SLI-021WO 30

[0180] F

[0181] 31

[0182] XXiXty™

[0183] M, N°

[0184] F

[0185] 32 F

[0186] N^°s^ O^OH

[0187] 33 F

[0188] (CXz

[0189] O"ZVA

[0190] 34

[0191] A?

[0192] 0'0 A

[0193] 35

[0194] OHFHOVO^^

[0195] Diastereomer 1

[0196] 36

[0197] 0 Xx

[0198] » VTJT\ JV™.

[0199]

[0200] Diastereomer 2

[0201] 16

[0202] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0203] 37

[0204] M yr- 38

[0205] O" T A

[0206] 39

[0207] VX

[0208] 40

[0209] O'V A

[0210] 41

[0211] O" T A

[0212] Diastereomer 1

[0213] 42 F

[0214] F O',

[0215] 4 A

[0216] Diastereomer 2

[0217] 43

[0218] MA f

[0219] H2NM\

[0220] f

[0221] 44

[0222] M T j

[0223] AM

[0224] W

[0225]

[0226] Diastereomer 1

[0227] 17

[0228] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0229] 45 F

[0230] F 0..

[0231] Diastereomer 2

[0232] 46 F

[0233] 47 F

[0234] 0F

[0235] 48 °->° V

[0236] ci( I n =

[0237] HiNyU

[0238] F

[0239] 49 F

[0240] O'T A

[0241] 50 F

[0242] i

[0243] X^yY^

[0244] of A

[0245] 51 F

[0246] X0y o

[0247] p Q ioh

[0248] O'T A

[0249] 52 F

[0250] F

[0251] H^0, Xy^

[0252]

[0253] O'ZT A

[0254] 18

[0255] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0256] 53 F

[0257] > — xA

[0258] O'o A

[0259] 54 F

[0260] O''T A

[0261] 55 XK F1

[0262] ^XcrA"

[0263] O / ZVA

[0264] 56

[0265] o'A A

[0266] 57

[0267] " X JX^[AOH

[0268] OZ'VA

[0269] 58 F

[0270] (T^t1

[0271] 59 oZzVX F

[0272] F 0^ JLAy'xN.N

[0273] HO"-^Q / ^QTX'0H

[0274] OZZVA

[0275]

[0276] Diastereomer 1

[0277] 19

[0278] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0279]

[0280] or a pharmaceutically acceptable salt thereof.

[0281] Definitions

[0282]

[0035] The term "aliphatic" or "aliphatic group", as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "carbocycle" "cycloaliphatic" or "cycloalkyl"), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of 20

[0283] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0284] attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl. An exemplary alkynyl group is an ethyne.

[0285]

[0036] The term "haloaliphatic" refers to an aliphatic group that is substituted with one or more halogen atoms.

[0286]

[0037] The term "haloalkyl" refers to a straight or branched alkyl group that is substituted with one or more halogen atoms.

[0287]

[0038] The term “alkyl” as used herein is a branched or unbranched saturated hydrocarbon group having a specified number of carbon atoms. As used herein, the term “alkyl” is understood to be an acyclic and fully saturated aliphatic group, which may be substituted or unsubstituted. In some embodiments, alkyl refers to a branched or unbranched saturated hydrocarbon group having three carbon atoms (C3). In some embodiments, alkyl refers to a branched or unbranched saturated hydrocarbon group having six carbon atoms (Ce). In some embodiments, the term “alkyl” includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, and hexyl.

[0288]

[0039] As used herein, the term “alkylene” refers to a bivalent alkyl group. An “alkylene chain” is a polymethylene group, i.e., — (CH2)n —, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group. The term "halogen" means F, Cl, Br, or I.

[0289]

[0040] The term "aryl" used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. The term "aryl" may be used interchangeably with the term "aryl ring". In certain embodiments of the present disclosure, "aryl" refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term "aryl", as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like.

[0290]

[0041] The terms "heteroaryl" and "heteroar-", used alone or as part of a larger moiety, e.g., "heteroaralkyl", or "heteroaralkoxy", refer to groups having 5 to 10 ring atoms, preferably 5,

[0291] 21

[0292] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0293] 6, or 9 ring atoms; having 6, 10, or 14 n electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms "heteroaryl" and "heteroar-", as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one. A heteroaryl group may be mono- or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring", "heteroaryl group", or "heteroaromatic", any of which terms include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0294]

[0042] As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4- dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or+NR (as in TV-substituted pyrrolidinyl). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic moiety", and "heterocyclic radical", are used interchangeably herein, and also include groups 22

[0295] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0296] in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring. A heterocyclyl group may be mono-or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0297]

[0043] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic moiety", and "heterocyclic radical", are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring. A heterocyclyl group may be mono-or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0298]

[0044] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation but is not intended to include aryl or heteroaryl moieties, as herein defined.

[0299]

[0045] As described herein, compounds of the disclosure may contain “optionally substituted” moieties. In general, the term “substituted”, whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0300] 23

[0301] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0302]

[0046] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group are independently halogen; — (CH2)o-4R°; — (CH2)o-40R°; — 0(CH2)o-4R°, — O— (CH2)O-4C(0)OR°; — (CH2)o-4CH(OR°)2; — (CH2)O-4SR°; — (CH2)o-4Ph, which may be substituted with R°; — (CH2)o-40(CH2)o-iPh which may be substituted with R°; — CH=CHPh, which may be substituted with R°; — (CH2)o-40(CH2)o-i-pyridyl which may be substituted with R°; — NO2; — CN; — N3; — (CH2)o-4N(R0)2; — (CH2)o-4N(R°)C(0)R°; — N(R°)C(S)R°; — (CH2)O-4N(R°)C(0)NR02; — N(R°)C(S)NR° 2; — (CH2)o-4N(R0)C(0)OR°; — N(R°)N(R°)C(O)R°; — N(R°)N(R°)C(O)NR° 2; — N(R’)N(R’)C(O)OR°; — (CH2)o-4C(0)R°; — C(S)R°; — (CH2)O-4C(0)OR°; — (CH2)O-4C(0)SR°; — (CH2)o-4C(0)OSiR° 3; — (CH2)o-4OC(O)R°; — OC(0)(CH2)O-4SR°, SC(S)SR°; — (CH2)O-4SC(0)R°; — (CH2)O-4C(0)NR° 2; — C(S)NR°2; — C(S)SR°; — SC(S)SR°, — (CH2)o-40C(0)NR° 2; — C(O)N(OR°)R°; — C(O)C(O)R°; — C(O)CH2C(O)R°; — C(NOR°)R°; — (CH2)o-4SSR°; — (CH2)o-4S(0)2R°; — (CH2)O-4S(0)20R°; — (CH2)O-40S(0)2R°; — S(O)2NR°2; — (CH2)O-4S(0)R“: — N(R°)S(O)2NR° 2; — N(R°)S(O)2R°; — S(O)(NR°)R°; — N(OR°)R°; — C(NH)NR° 2; — P(O)2R°; — P(O)R° 2; — OP(O)R° 2; — OP(O)(ORo)z; SiR° 3; — (C1-4 straight or branched alkylene)O — N(R°)2; or — (C1-4 straight or branched alkylene)C(O)O — N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, — CFhPh, — 0(CH2)o iPh, — CH2-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0303]

[0047] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, — (CH2)o-2R*, -(haloR*), — (CH2)O-20H, — (CH2)O-20R‘, — (CH2)O-2CH(OR*)2; — O(haloR*), — CN, — N3, — (CH2)O-2C(0)R*, — (CH2)O-2C(0)OH, — (CH2)O-2C(0)OR‘, — (CH2)O-2SR*, — (CH2)O- 2SH, — (CH2)O-2NH2, — (CH2)O-2NHR*, — (CH2)O-2NR* 2, — NO2, — SiR* 3, — OSiR* 3, — C(O)SR*, — (C1-4 straight or branched alkylene)C(O)OR*, or — SSR* wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently selected from C1-4 aliphatic, — CFhPh, — 0(CH2)o iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of

[0304] 24

[0305] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0306] R° include =0 and =S.

[0307]

[0048] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted’’ group include the following: =0, =S, =NNR*2, =NNHC(0)R*, =NNHC(0)0R*, =NNHS(O)2R*, =NR*, =N0R*, — O(C(R*2))2-3O— or — S(C(R*2))2-3S—, wherein each independent occurrence of R* is selected from hydrogen, Ci-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: — O(CR*2)2-3O —, wherein each independent occurrence of R* is selected from hydrogen, Ci-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0308]

[0049] Suitable substituents on the aliphatic group of R* include halogen, — R*, -(haloR*), — OH, — OR*, — O(haloR*), — CN, — C(O)OH, — C(O)OR*, — NH2, —NHR*, —NR* 2, or — NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently Ci-4 aliphatic, — CH2Ph, — 0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0309]

[0050] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include — R\ — NRf2, — C(O)Rf, — C(O)OR\ — C(O)C(O)Rt, — C(O)CH2C(O)Rt, — S(O)2Rf, — S(O)2NRf2, — C(S)NRt2, — C(NH)NRf2, or — N(Rt)S(O)2Rt; wherein each R' is independently hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted — OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R'. taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0310]

[0051] Suitable substituents on the aliphatic group of R1are independently halogen, — R*, -(haloR*), —OH, —OR*, — O(haloR*), — CN, — C(O)OH, — C(O)OR*, — NH2, —NHR*, — NR* 2, or — NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, — CH2Ph, — 0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0311] 25

[0312] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0313]

[0052] As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are 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, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include 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-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.

[0314]

[0053] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N(Ci-4alkyl)4 salts. 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, lower alkyl sulfonate and aryl sulfonate.

[0315]

[0054] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0316]

[0055] The term "biological sample", as used herein, includes, without limitation, cell cultures or extracts thereof: biopsied material obtained from a mammal or extracts thereof: and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. Examples of such 26

[0317] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0318] purposes include, but are not limited to, blood transfusion, organ transplantation, biological specimen storage, and biological assays.

[0319]

[0056] As used herein, a "therapeutically effective amount" means an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount that is sufficient, when administered as part of a dosing regimen to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat and / or diagnose the onset of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount of a substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of a provided compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, reduces severity of and / or reduces incidence of one or more symptoms or features of the disease, disorder, and / or condition. In some embodiments, a "therapeutically effective amount" is at least a minimal amount of a provided compound, or composition containing a provided compound, which is sufficient for treating one or more symptoms of an CFTR-associated disease or disorder.

[0320]

[0057] The terms “treat”, “treatment” or “treating” mean to decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease (e.g., a disease or disorder delineated herein), lessen the severity of the disease or improve the symptoms associated with the disease. Treatment includes treating a symptom of a disease, disorder or condition. Without being bound by any theory, in some embodiments, treating includes augmenting deficient CFTR activity. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the subject) then the treatment is prophylactic (i.e., it protects the subject against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic, (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof).

[0321]

[0058] The term "subject" to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys. Preferred subjects are humans.

[0322] 27

[0323] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0324]

[0059] The term “pharmaceutically acceptable carrier, adjuvant, or vehicle” refers to a nontoxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound(s) with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of the compounds disclosed herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0325]

[0060] A “pharmaceutically acceptable derivative” means any non-toxic salt, ester, salt of an ester or other derivative of a compound of this disclosure that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this disclosure or an active metabolite or residue thereof.

[0326]

[0061] The expression “dosage unit form” as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that total daily usage of compounds and compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. Specific effective dose level for any particular patient or organism will depend upon a variety of factors including disorder being treated and severity of the disorder; activity of specific compound employed; specific composition employed; age, body weight, general health, sex and diet of the patient; time of administration, route of administration, and rate of excretion of a specific compound employed; duration of treatment; drugs used in combination or coincidental with a specific compound employed, and like factors well known in the medical arts.

[0327]

[0062] A “response” to a method of treatment can include a decrease in or amelioration of negative symptoms, a decrease in the progression of a disease or symptoms thereof, an increase in beneficial symptoms or clinical outcomes, a lessening of side effects, stabilization of disease, partial or complete remedy of disease, among others.

[0328]

[0063] As used herein, “CFTR” means cystic fibrosis transmembrane conductance regulator. Defects in the function of the CFTR ion channel result from loss of function mutations of CFTR. Such mutations lead to exocrine gland dysfunction, abnormal mucociliary clearance, and cause cystic fibrosis. The most common CFTR mutation in Cystic Fibrosis (CF) patients 28

[0329] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0330] leads to the specific deletion of three nucleotides of the codon for phenylalanine at position 508. This mutation, which is found in -70% of CF patients worldwide, is referred to as “AF508”. The AF508 mutation decreases the stability of the CFTR NBD1 domain and limits CFTR interdomain assembly. Since CF is an autosomal recessive disease, a CF patient harboring the AF508 CFTR mutation must also carry a second defective copy of CFTR. Approximately 2000 different CF-causing CFTR mutations have been identified in CF patients. CF patients harboring the AF508 CFTR mutation can be homozygous for that mutation (AF508 / AF508). CF patients can also be AF508 heterozygous, if the second CFTR allele such patients carry instead contains a different CFTR loss of function mutation. Such CFTR mutations include, but are not limited to, G542X, G551D, N13O3K, W1282X, R553X, R117H, R1162X. R347P, G85E, R560T, A455E, AI507, G178R, S549N, S549R, G551S, G970R, G1244E, S1251N, S1255P, and G1349D.

[0331]

[0064] As used herein, the term “CFTR modulator” refers to a compound that increases the activity of CFTR. In certain aspects, a CFTR modulator is a CFTR corrector or a CFTR potentiator or a dual-acting compound having activities of a corrector and a potentiator.

[0332]

[0065] As used herein, the term “CFTR corrector” refers to a compound that increases the amount of functional CFTR protein to the cell surface and thus enhances CFTR channel function. The CFTR correctors partially “rescue” misfolding of CFTR, thereby enabling the maturation and functional expression of CFTR protein harboring a CF causing mutation on the cell surface. Examples of correctors include, but are not limited to, VX-809, VX-661, VX-152, VX-440, VX-983, and GLPG2222. Such compounds may interact directly with CFTR protein, modifying its folding and conformational maturation during synthesis.

[0333]

[0066] As used herein, the term “CFTR potentiator” refers to a compound that increases the ion channel activity of CFTR protein located at the cell surface, resulting in enhanced ion transport. CFTR potentiators repair the defective channel functions caused by mutations. Examples of potentiators include, but are not limited to, ivacaftor (VX770), deuterated ivacaftor (CPT 656), genistein and GLPG1837.

[0334]

[0067] As used herein, the term “CFTR pharmacological chaperone” (PC) refers to compounds that stabilize the CFTR protein in its native state by binding directly to the protein.

[0335]

[0068] As used herein, the term “CFTR proteostasis regulator” (PR) refers to compounds that enhance the protein folding efficiency within the cell. PRs can alter the activity of transcriptional, folding and / or membrane trafficking machinery, as well as impeding the degradation of partially folded, but functional, conformers at the endoplasmic reticulum (ER) or plasma membrane.

[0336] 29

[0337] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0338]

[0069] As used herein, “CFTR disease or condition" refers to a disease or condition associated with deficient CFTR activity, for example, cystic fibrosis, congenital bilateral absence of vas deferens (CBAVD), acute, recurrent, or chronic pancreatitis, disseminated bronchiectasis, asthma, allergic pulmonary aspergillosis, smoking -related lung diseases, such as chronic obstructive pulmonary disease (COPD), chronic sinusitis, dry eye disease, protein C deficiency, A-beta.-lipoproteinemia, lysosomal storage disease, type 1 chylomicronemia, mild pulmonary disease, lipid processing deficiencies, type 1 hereditary angioedema, coagulationfibrinolysis, hereditary hemochromatosis, CFTR-related metabolic syndrome, chronic bronchitis, constipation, pancreatic insufficiency, hereditary emphysema, and Sjogren's syndrome.

[0339]

[0070] As used herein, the term "combination," "combined," and related terms refers to the simultaneous or sequential administration of therapeutic agents in accordance with this disclosure. For example, a compound of the present disclosure may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present disclosure provides a single unit dosage form comprising a provided compound, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0340]

[0071] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and 5 configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. A person of ordinary skill in the art will understand that tautomeric forms of the compounds disclosed herein are within the scope of the present disclosure.

[0341] Alternative Embodiments

[0342]

[0072] In an alternative embodiment, compounds described herein may also comprise one or more isotopic substitutions. For example, hydrogen may be2H (D or deuterium) or3H (T or tritium); carbon may be, for example,13C or14C; oxygen may be, for example,18O; nitrogen may be, for example,15N, and the like. In other embodiments, a particular isotope (e.g.,3H,13C,14C,18O, or15N) can represent at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 30

[0343] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0344] 95%, at least 99%, or at least 99.9% of the total isotopic abundance of an element that occupies a specific site of the compound.

[0345] Pharmaceutical Compositions

[0346]

[0073] In some embodiments, the present disclosure provides a composition comprising a compound of the present disclosure and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In some embodiments, the amount of compound in compositions contemplated herein is such that is effective to measurably modulate CFTR, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, the amount of compound in compositions of this disclosure is such that is effective to measurably modulate CFTR, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, a composition contemplated by this disclosure is formulated for administration to a patient in need of such composition. In some embodiments, a composition contemplated by this disclosure is formulated for oral administration to a patient.

[0347]

[0074] In some embodiments, the amount of compound in compositions contemplated herein is such that is effective to measurably modulate a protein, particularly at CFTR, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, the amount of compound in compositions of this disclosure is such that is effective to measurably modulate CFTR, or a mutant thereof, in a biological sample or in a patient.

[0348]

[0075] In some embodiments, compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. In some preferred embodiments, compositions are administered orally, intraperitoneally or intravenously. In some embodiments, sterile injectable forms of the compositions comprising one or more compounds of the present disclosure may be aqueous or oleaginous suspension. In some embodiments, suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. In some embodiments, sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. In some embodiments, among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In some embodiments, additional examples include, but are not limited to, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0349]

[0076] The term “parenteral” as used herein includes subcutaneous, intravenous,

[0350] 31

[0351] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0352] intramuscular, intra-articular, intra-synovial, intra-sternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.

[0353]

[0077] Pharmaceutically acceptable compositions comprising one or more compounds of the present disclosure may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In some embodiments, carriers used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. In some embodiments, useful diluents include lactose and dried cornstarch. In some embodiments, when aqueous suspensions are required for oral use, an active ingredient is combined with emulsifying and suspending agents. In some embodiments, certain sweetening, flavoring or coloring agents may also be added.

[0354]

[0078] Alternatively, pharmaceutically acceptable compositions comprising a compound of the present disclosure may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0355]

[0079] Pharmaceutically acceptable compositions comprising a compound of the present disclosure may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs. In some embodiments, pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0356]

[0080] Pharmaceutically acceptable compositions comprising a compound of the present disclosure may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives,

[0357] 32

[0358] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0359] absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0360]

[0081] In some embodiments, an amount of a compound of the present disclosure that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of between 0.01-100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0361] Methods of Using Compounds of the Present Disclosure

[0362]

[0082] As discussed above, CFTR is composed of two six membrane-spanning domains (MSD1 and MSD2), two nucleotide bind domains (NBD1 and NBD2), a regulatory region (R) and four cytosolic loops (ICL-4). CFTR protein is located primarily in the apical membrane of epithelial cells where it functions to conduct anions, including chloride, bicarbonate, and thiocyanate into and out of the cell. The most frequent CFTR mutation is the in-frame deletion of phenylalanine at residue 508 (AF508) in the first nucleotide binding domain (NBD1). The mutation has several deleterious effects on the production of CFTR in the ER, its correct folding, its movement to the plasma membrane and its normal function as an ion channel for the cell.

[0363]

[0083] One such negative effect is that the NBD1 domain is partially or mis-folded which is recognized within the cell as an aberrant protein and tagged for disposal by ER-associated degradation (ERAD) via the ubiquitin-proteasome system (UPS). Should a partially or misfolded CFTR protein emerge from the ER, the protein must travel to the plasma membrane through complex glycosylation in the Golgi compartment and be functionally inserted. In wildtype CFTR, only 20-40% of CFTR reaches the plasma membrane, indicating that CFTR has energetic instability of individual NBDs, a slow domain assembly, and relatively fast ERAD kinetics which all contribute to inefficient folding and sensitize CFTR to structural perturbations by mutations.

[0364]

[0084] In wild-type CFTR, the NBD1 domain folds co-translationally while other domains fold post-translationally. Mutated AF508 CFTR has impaired NBD1 folding but its backbone structure and thermodynamic stability are similar to wild-type CFTR. With delayed folding kinetics, mutated AF508 CFTR NBD1 has an increased folding activation energy. Lack of proper folding results in hydrophobic residues being exposed to the surface of NBD1 which causes aggregation with other CFTR proteins. Thus, the aggregation temperature of mutated

[0365] 33

[0366] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0367] CFTR drops from 41 °C to 33 °C. This level of instability creates a greater percentage of misfolded mutant CFTR at physiological temperature (37 °C in humans). Mutant CFTR suffers from both kinetic and thermodynamic folding defects. CFTR stabilizers can address these folding defects, but complete energetic correction of mutant NBD1 folding has been shown to not result in the CFTR biosynthetic processing, underscoring the need for interface stability as well.

[0368]

[0085] The disclosed CFTR correctors can interact with the NBD domain to stabilize the correct folded position R, such that CFTR is not labeled for elimination from the cell. The preservation of correct folding enables CFTR to function as a chloride ion channel at wild-type levels. In some embodiments, disclosed CFTR correctors can enhance the performance of wild-type CFTR.

[0369]

[0086] CFTR stabilizers can function in combination with other therapeutic agents such as CFTR correctors that promote A508 CFTR exit from the ER and accumulation in the plasma membrane. Increasing the amount of CFTR cell surface expression can result in improved chloride conductance following channel activation by both potentiators and a cAMP agonist. Thus, disclosed herein are combinations of CFTR stabilizers with CFTR correctors and potentiators, optionally with cAMP agonists or another therapeutic agent as described below.

[0370]

[0087] Disclosed herein are methods of treating deficient CFTR activity in a cell, comprising contacting the cell with a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. In certain embodiments, contacting the cell occurs in a subject in need thereof, thereby treating a disease or disorder mediated by deficient CFTR activity.

[0371]

[0088] Also, disclosed herein are methods of treating a disease or a disorder mediated by deficient CFTR activity comprising administering a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a mammal, preferably a human. In some embodiments, the disease is associated with the regulation of fluid volumes across epithelial membranes, particularly an obstructive airway disease such as CF or COPD.

[0372]

[0089] Such diseases and conditions include, but are not limited to, cystic fibrosis, asthma, smoke induced COPD, chronic bronchitis, rhinosinusitis, constipation, pancreatitis, pancreatic insufficiency, male infertility caused by congenital bilateral absence of the vas deferens (CBAVD), mild pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, familial hypercholesterolemia, Type 1 chylomicronemia,

[0373] 34

[0374] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0375] abetalipoproteinemia, lysosomal storage diseases, I-cell disease / pseudo-Hurler, mucopolysaccharidoses, Sandhof / Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy / hyperinsulinemia, Diabetes mellitus, Laron dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), neurophyseal DI, neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, several polyglutamine neurological disorders, Huntington's, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, myotonic dystrophy, spongiform encephalopathies, hereditary Creutzfeldt- Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, Sjogren's disease, Osteoporosis, Osteopenia, bone healing and bone growth, bone repair, bone regeneration, reducing bone resorption, increasing bone deposition, Gorham's Syndrome, chloride channelopathies, myotonia congenita, Bartter's syndrome type III, Dent's disease, hyperekplexia, epilepsy, hyperekplexia, lysosomal storage disease, Angelman syndrome, Primary Ciliary Dyskinesia (PCD), PCD with situs inversus, PCD without situs inversus and ciliary aplasia.

[0376]

[0090] Such diseases and conditions include, but are not limited to, cystic fibrosis, congenital bilateral absence of vas deferens (CBAVD), acute, recurrent, or chronic pancreatitis, disseminated bronchiectasis, asthma, allergic pulmonary aspergillosis, chronic obstructive pulmonary disease (COPD), chronic sinusitis, dry eye disease, protein C deficiency, Abetalipoproteinemia, lysosomal storage disease, type 1 chylomicronemia, mild pulmonary disease, lipid processing deficiencies, type 1 hereditary angioedema, coagulation-fibrinolysis, hereditary hemochromatosis, CFTR-related metabolic syndrome, chronic bronchitis, constipation, pancreatic insufficiency, hereditary emphysema, and Sjogren's syndrome. In some embodiments, a disease is cystic fibrosis.

[0377]

[0091] Provided herein are methods of treating cystic fibrosis, comprising administering to a subject in need thereof, a compound as disclosed herein or a pharmaceutically acceptable salt thereof. Also provided herein are methods of lessening the severity of cystic fibrosis, comprising administering to a subject in need thereof, a compound as disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a human. In some embodiments, the subject is at risk of developing cystic fibrosis, and administration is carried out prior to the onset of symptoms of cystic fibrosis in the subject.

[0378]

[0092] Provided herein are compounds as disclosed herein for use in treating a disease or 35

[0379] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0380] condition mediated by deficient CFTR activity. Also provided herein are uses of a compound as disclosed herein for the manufacture of a medicament for treating a disease or condition mediated by deficient CFTR activity.

[0381]

[0093] Provided herein are kits for use in measuring the activity of CFTR or a fragment thereof in a biological sample in vitro or in vivo. The kit can contain: (i) a compound as disclosed herein, or a pharmacal composition comprising the disclosed compound, and (ii) instructions for: a) contacting the compound or composition with the biological sample; and b) measuring activity of said CFTR or a fragment thereof. In some embodiments, the biological sample is biopsied material obtained from a mammal or extracts thereof; blood, saliva, urine, feces, semen, tears, other body fluids, or extracts thereof. In some embodiments, the mammal is a human.

[0382]

[0094] Provided herein are compounds as disclosed herein for use in treating kidney disease. In some embodiments, a kidney disease is autosomal dominant polycystic kidney disease (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD). In some embodiments, a kidney disease is autosomal dominant polycystic kidney disease (ADPKD). In some embodiments, a kidney disease is autosomal recessive polycystic kidney disease (ARPKD).

[0383] Combination Treatments

[0384]

[0095] As used herein, the term "combination therapy" means administering to a subject (e.g., human) two or more CFTR modulators, or a CFTR modulator and an agent such as antibiotics, ENaC inhibitors, GSNO (S-nitrosothiol, s-nitroglutathione) reductase inhibitors, and a CRISPR Cas correction therapy or system (as described in US 2007 / 0022507 and the like). In some embodiments, combination therapy includes administration of a compound described herein with a compound that modulates CFTR protein or ABC protein activities (e.g., as described in WO2018167690A1 and the like).

[0385]

[0096] In certain embodiments, the method of treating a disease or condition mediated by deficient CFTR activity comprises administering a compound as disclosed herein conjointly with one or more other therapeutic agent(s). In some embodiments, one other therapeutic agent is administered. In other embodiments, at least two other therapeutic agents are administered.

[0386]

[0097] In certain embodiments, the method of preventing a disease or condition mediated by deficient CFTR activity comprises administering a compound as disclosed herein conjointly with one or more other therapeutic agent(s). In some embodiments, one other therapeutic agent is administered. In other embodiments, at least two other therapeutic agents are administered.

[0387] 36

[0388] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0389]

[0098] Additional therapeutic agents include, for example, ENaC inhibitors, mucolytic agents, modulators of mucus rheology, bronchodilators, antibiotics, anti-infective agents, antiinflammatory agents, ion channel modulating agents, therapeutic agents used in gene or mRNA therapy, agents that reduce airway surface liquid and / or reduce airway surface PH, CFTR correctors, and CFTR potentiators, or other agents that modulate CFTR activity. Other therapeutics include liposomal composition components such as those described in WO2012 / 170889, hybrid oligonucleotides that facilitate RNA cleavage such as those described in WO2016 / 130943, and single stranded oligonucleotides that modulate gene expression as described in WO2016 / 130929.

[0390]

[0099] In some embodiments, at least one additional therapeutic agent is selected from one or more CFTR modulators, one or more CFTR correctors and one or more CFTR potentiators.

[0391]

[0100] Non-limiting examples of additional therapeutics include VX-770 (Ivacaftor), VX-809 (Eumacaftor, 3-(6-(I-(2,2-5 difluorobenzo[d][l, 3]dioxol-5-yl)cyclopropanecarboxamido)-3-methylpyridin-2-yl) benzoic acid, VX-661 (Tezacaftor, I-(2,2-difluoro-l, 3-benzodioxol-5-yl)-N-[I-[(2R)-2,3-dihydroxypropyl]-6-fluoro-2-(2-hydroxy-l, I-dimethylethyl)- IH-indol-5-yl]- cyclopropanecarboxamide), VX-983, VX-152, VX-440, VX-445, VX-659, VX-371. Orkambi, Ataluren (PTC 124) (3-[5-(2-fluorophenyl)-l, 2,4-oxadiazol-3-yl]benzoic acid), PTI-130 (Proteostasis), PTI-801, PTI-808, PTI-428, N91115.74 (cavosonstat), QBW251 (Novartis) compounds described in WO2011113894, compounds N30 Pharmaceuticals (e.g., WO 2014 / 186704), deuterated ivacaftor (e.g., CTP-656 or VX-561), GEPG 2222, GEPG2451, GEPG3067, GEPG2851, GEPG2737, GEPG 1837 (N-(3-carbamoyl-5,5,7,7-tetramethyl-5,7-dihydro-4H-thieno[2,3-c]pyran-2-yl)-lH-pyrazole-5-carboxamide), GEPG 2665 (Galapagos), FDE 169 (Flatley Discovery lab), FDE 176, FDE438, FDE304, FD2052160, FD1881042, FD2027304, FD2035659, FD2033129, FD1860293, CFFT-PotOl, CFFT-Pot-02, P-1037, glycerol, phenylbutyrate, and the like.

[0392]

[0101] Non-limiting examples of additional therapeutics include compounds disclosed in US Patent Application Nos. PCT / US20 / 63586, PCT / US20 / 63589, and PCT / US 20 / 63590, each of which is incorporated by reference in its entirety.

[0393]

[0102] Non-limiting examples of anti-inflammatory agents are N6022 (3-(5-(4-(IH-imidazol-I-yl)10 phenyl)-I-(4-carbamoyl-2-methylphenyl)-'H-pyrrol-2-yl) propanoic acid), Ibuprofen, Eenabasum (anabasum), Acebilustat (CTX-4430), LAU-7b, POL6014, docosahexaenoic acid, alpha-1 anti-trypsin, sildenafil. Additional therapeutic agents also include, but are not limited to a mucolytic agent, a modifier of mucus rheology (such as hypertonic saline, mannitol, and oligosaccharide based therapy), a bronchodilator, an anti -infective (such as tazobactam,

[0394] 37

[0395] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0396] piperacillin, rifampin, meropenem, ceftazidime, aztreonam, tobramycin, fosfomycin, azithromycin, amitriptyline, vancomycin, gallium and colistin), an anti-infective agent, an antiinflammatory agent, a CFTR modulator other than a compound of the present disclosure, and a nutritional agent. Additional therapeutic agents can include treatments for comorbid conditions of cystic fibrosis, such as exocrine pancreatic insufficiency which can be treated with Pancrelipase or Liprotamase.

[0397]

[0103] Examples of CFTR potentiators include, but are not limited to, Ivacaftor (VX-770), CTP-656, NVS-QBW251, FD1860293, GLPG2451, GLPG1837, and N-(3-carbamoyl-5,5,7,7-tetramethyl-5,7-dihydro-4H-thieno[2,3-c]pyran-2-yl)-lH-pyrazole-5-carboxamide. Examples of potentiators are also disclosed in publications: W02005120497, WO2008147952, W02009076593, W02010048573, WG2006002421, WO2008147952, W02011072241, WO2011113894, WO2013038373, WO2013038378, WO2013038381, WO2013038386, W02013038390, WO2014180562, WO2015018823, and U. S. patent application Ser. Nos.

[0398] 14 / 271,080, 14 / 451,619 and 15 / 164,317.

[0399]

[0104] Non-limiting examples of correctors include Lumacaftor (VX-809), l-(2,2-difluoro-l,3-benzodioxol-5-yl)-N-{ l-[(2R)-2,3-dihydroxypropyl]-6-fluoro-2-(l-hydroxy-2-methylpropan-2-yl)- 1 H-indol-5 -yl } cyclopropanecarboxamide ( VX-661 ), VX-983, GLPG2222, GLPG2665, GLPG2737, VX-152, VX-440, FDL169, FDL304, FD2052160, and FD2035659. Examples of correctors are also disclosed in US20160095858A1, and U. S. application Ser. Nos. 14 / 925,649 and 14 / 926,727.

[0400]

[0105] In certain embodiments, an additional therapeutic agent is a CFTR amplifier. CFTR amplifiers enhance the effect of known CFTR modulators, such as potentiators and correctors. Examples of CFTR amplifier include PTI130 and PTI-428. Examples of amplifiers are also disclosed in publications: WO2015138909 and WO2015138934.

[0401]

[0106] In certain embodiments, the additional therapeutic agent is an agent that reduces the activity of the epithelial sodium channel blocker (ENaC) either directly by blocking the channel or indirectly by modulation of proteases that lead to an increase in ENaC activity (e.g., serine proteases, channel-activating proteases). Exemplary of such agents include camostat (a trypsinlike protease inhibitor), QAU145, 552-02, GS-9411, INO-4995, Aerolytic, amiloride, AZD5634, and VX-371. Additional agents that reduce the activity of the epithelial sodium channel blocker (ENaC) can be found, for example, in PCT Publication No. W02009074575 and W02013043720; and U. S. Pat. No. 8,999,976.

[0402]

[0107] In some embodiments, an additional therapeutic agent is a compound disclosed in publications: WO2023034946, W02024054840, W02021097054, W02021097057,

[0403] 38

[0404] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0405] WO2021113806, WO2021113808, WO2021113809, WO2023034992, WO2024054845, WO2024054851, WO2018167690, WO2023240267. WO2024182736, WO2024182739, WO2024182742, WO2024182751, WO2016069757, W02017009804, and W02017208115, the entirety of each of which are incorporated in their entirety.

[0406]

[0108] In one embodiment, an ENaC inhibitor is VX-371.

[0407]

[0109] In one embodiment, an ENaC inhibitor is SPX-101 (S18).

[0408]

[0110] In certain embodiments, the combination of a compound of the present disclosure, with a second therapeutic agent may have a synergistic effect in the treatment of cancer and other diseases or disorders mediated by adenosine. In other embodiments, the combination may have an additive effect.

[0409] Exemplification

[0410] Abbreviations:

[0411] Boc: tert-butyloxy carbonyl

[0412] DEA: diethyl amine

[0413] DBU: l,8-Diazabicyclo[5.4.0]undec-7-ene

[0414] DMSO: dimethyl sulfoxide

[0415] dppf: 1, 1 '-Bis(diphenylphosphino)ferrocene

[0416] DTT: dithiothreitol

[0417] ESI: electron spray ionization

[0418] ee: enantiomeric excess

[0419] HATU: l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0420] HPLC: high performance liquid chromatography

[0421] LC-MS: liquid chromatography-mass spectrometry

[0422] m-CPBA: meta-chloroperbenzoic acid

[0423] NBS: A-bromosuccinimide

[0424] NCS: A-chlorosuccinimide

[0425] NIS: A-iodosuccinimide

[0426] Pd / C: Palladium on carbon

[0427] RT: retention time

[0428] SFC: supercritical fluid chromatography

[0429] TBS: / c 7- Butyldi methylsilyl

[0430] 39

[0431] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0432] TIPS: Triisopropylsilyl

[0433] THF: tetrahydrofuran

[0434] THP: tetrahydropyran

[0435] Ts: tosyl

[0436] General formula:

[0437]

[0438] General Procedures

[0439]

[0111] The compounds of the present disclosure can be better understood in connection with the following synthetic schemes and methods which illustrate means by which the compounds of the Formula (I) can be prepared. The compounds of this disclosure can be prepared by a variety of synthetic procedures illustrated in Schemes 1 to 13.

[0440] Scheme 1: Thio-anilino-nitrile synthesis

[0441] reduction

[0442] Br

[0443]

[0444] S-1 S-3 S-4

[0445]

[0112] The thiol S-l was subjected to SNAr reaction with S-2 (Ml, M2, M3, M4 = N, CH, CF, CO, CBr, CI) resulting in nitro intermediate S-3, which was reduced to an aniline by reducing agents such as iron / acetic acid, zinc / ammonium chloride, or catalytic hydrogenation.

[0446] Scheme 2: Thio-phenol-nitrile synthesis

[0447] protection

[0448]

[0449] S1-4 S2-1 S2-2

[0450]

[0113] The aniline Sl-4 was converted into phenol S2-lvia diazonium formation followed by hydroxylation. The resulting phenol was then protected to generate the nitrile S2-2.

[0451] 40

[0452] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0453] Scheme 3: Alcohol-nitrile synthesis.

[0454] hydroxyl

[0455] group

[0456] protection

[0457] Br OH Br

[0458]

[0459] S3-1 S3-3 S3-4

[0460]

[0114] Deprotonation of S3-1 followed by addition of aldehyde S3-2 formed the alcohol S3-3. The alcohol thus formed was protected as nitrile S3 -4.

[0461] Scheme 4: Thio-thiazole aniline synthesis.

[0462]

[0463] S1-1 S4-2 S4-3 S4-4

[0464]

[0115] The thiol Sl-1 was subjected to SNAr with S4-1 to form thioether S4-2. The aniline was bis-protected (such as with Boc) to afford S4-3. Bromination of the thiazole with NBS, bromine or other bromination reagent affords nitrile S4-4.

[0465] Scheme 5: Aniline-nitrile synthesis.

[0466]

[0467] S5-6 S5-5

[0468]

[0116] When the aniline substitution pattern was not suitable for the SNAr substitution shown, as in Scheme 1, as an alternative, an iodonium coupling reaction was adopted. Phenol S5-1 was reacted with S5-2 to afford biarylether S5-3, which was then converted to nitrile S5-6 via a Buchwald coupling reaction.

[0469] Scheme 6: Synthesis of imidothioate and triazole formation

[0470] H

[0471]

[0472] S6-2 S6-3 S6-5

[0473] 41

[0474] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0475]

[0117] Nitriles like S6-1 were converted to thioamides S6-2, using sodium hydrogen sulfide / magnesium sulfate, or related methods. The thioamide was then used to prepare the imidothioate S6-3. Reaction of imidothioate S6-3 with hydrazide S6-4 afforded triazole S6-5.

[0476] Scheme 7: Synthesis of vinyl bromide macrocyclic precursor.

[0477] nitro reduction

[0478] aniline protection

[0479]

[0480]

[0118] One of the ways to make the vinyl bromide macrocyclic precursor (S7-6) is presented in Scheme 7. SNAr substitution of para-fluoro-nitro compound S7-2 with phenol S7-1 gave nitro compound S7-3. Dehydration of 2-hydroxysulfone afforded alkene S7-4. The nitro group was reduced with reducing agents like iron, zinc, and the like. The aniline group thus obtained was bis-protected with protecting groups such as Boc.

[0481] Scheme 8. Synthesis of pyridyl anilines.

[0482]

[0483]

[0119] The pyridyl aniline analogues were prepared in a similar manner as described in Scheme 7. Reaction of para- or ortho- fluoro substituted pyridine S8-2 with phenol S8-1 resulted in alcohol S8-3. Dehydration of alcohol S8-3 resulted in vinyl bromide S8-4.

[0484] 42

[0485] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0486] Scheme 9. Macrocyclization.

[0487]

[0488]

[0120] The vinyl bromide S9-1 was subjected to intramolecular Heck coupling conditions, involving a catalyst like tri-tert-butylphosphine palladium, to form macrocyclic S9-2. The double bond of S9-2 was reduced via catalytic hydrogenation or with reducing reagents like tosyl hydrazide to afford S9-3.

[0489] Scheme 10. Halogenation of aromatic aniline.

[0490]

[0491]

[0121] Aniline S 10-1 was halogenated with NCS, NIS, NBS, bromine or other halogenation reagents to afford S10-2.

[0492] Scheme 11. Ketone-bridged analogue synthesis.

[0493]

[0122] The ketone analogues were obtained by oxidation of alcohol S 11 -2 to the correspondin; ketone SI 1-3.

[0494]

[0495] 43

[0496] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0497] Scheme 12-1. Sulfoxide synthesis.

[0498]

[0499]

[0123] The sulfur-bridged macrocyclic compound S 12- 1 was oxidized to sulfoxide and sulfone using oxidants like hydrogen peroxide, m-CPBA, and the like.

[0500] Scheme 13: Alternative macrocyclization strategy

[0501]

[0502]

[0124] Another macrocyclization method is presented in Scheme 13. Heck reaction of vinyl sulfone S13-1 with bromide S13-2, followed by catalytic hydrogenation, affords sulfone SI 3- 3. Bromination of thiazole S13-3 and deprotection of the aniline affords phenol bromide SI 3- 4. The macrocyclization is accomplished in the presence of bases, such as sodium carbonate, to afford SI 3-5.

[0503] Analytical Methods:

[0504]

[0125] Analytical Procedures1H NMR spectra were recorded with a Bruker AC 400 MHz apparatus. Chemical shifts (8) are quoted in parts per million (ppm) and coupling constants (J) in hertz (Hz).

[0505]

[0126] The following liquid chromatography-mass spectrometry (LC-MS) methods were used. LC-MS Method 1.

[0506]

[0127] Mobile Phase: A: water (0.1 % formic acid) B: acetonitrile (0.1 % formic acid), Gradient: 5% B 0.2 minutes, increase to 95% B within 1.3 minutes, 95% B for 1.5 minutes, back to 5% B within 0.01 minutes; Flow Rate: 2 mL / min; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Oven Temperature: 50 °C; Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1000 amu).

[0507] 44

[0508] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0509] LC-MS Method 2.

[0510]

[0128] Mobile phase: A: water (0.01% trifluoracetic acid), B: acetonitrile (0.01% trifluoracetic acid); Gradient: 5 to 95% of B in 1.4 minutes, 95% B for 1.6 minutes, back to 5% B within 0.01 minutes; Flow Rate: 2.0 ml / minutes; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Temperature: 50 °C; Detection: UV (214 nm, 4 nm; 254 nm, 4 nm) and MS (ESI, Pos mode, 110 to 1000 amu).

[0511] LC-MS Method 3.

[0512]

[0129] Mobile phase: A: water (10% ammonium bicarbonate), B: acetonitrile; Gradient: 10 to 95% of B in 1.5 minutes, 95% B for 1.5 minutes, back to 10% B within 0.01 minutes; Flow Rate: 1.8 ml / minutes; Column: XBRIGE C18, 4.6 x 50 mm, 3.5 pm; Temperature: 50 °C; Detection: UV (214 nm, 4 nm; 254 nm, 4 nm) and MS (ESI, Pos mode, 103 to 1500 amu). LC-MS Method 4:

[0513]

[0130] Mobile Phase: A: water (0.01% trifluoroacetic acid) B:: acetonitrile (0.01% trifluoroacetic acid); Gradient: from 5% to 95% B in 2.5 minutes, 95% B for 2.5 minutes, back to 5% B 0.01 minutes; Flow Rate: 2 ml / minute; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Oven Temperature: 50 °C; Detection: UV (214 nm, 4 nm; 254 nm, 4 nm) and MS (ESI, Pos mode, 120 to 1500 amu).

[0514] LC-Mass Method 5:

[0515]

[0131] Mobile Phase: A: water (0.01% trifluoroacetic acid) B:: acetonitrile (0.01% trifluoroacetic acid); Gradient: 5% increase to 95%B within 1.3 minutes, 95%B for 1.7 minutes, back to 5% B 0.01 minutes; Flow Rate: 1.8 ml / minute: Column: Sunfire Cl 8, 4.6 x 50 mm, 3.5 pm; Column Temperature: 50 °C. Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 50 to 900 amu).

[0516] LC-Mass Method 6:

[0517]

[0132] Mobile Phase: A: water (0.01% trifluoroacetic acid) B: acetonitrile (0.01% trifluoroacetic acid); Gradient: 5% increase to 95%B in 3 minutes, 95%B for 2 minutes, back to 5% B 0.01 minutes; Flow Rate: 2.0 ml / minute; Column: HALO C18, 4.6 x 30 mm, 2.7 pm; Column Temperature: 50 °C. Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1200 amu).

[0518] LC-MS Method 7.

[0519]

[0133] Mobile phase: A: water (0.01% trifluoroacetic acid) B: acetonitrile (0.01% trifluoroacetic acid); Gradient: 5 to 95% of B in 1.5 minutes, 95% B for 1.5 minutes, back to

[0520] 45

[0521] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0522] 5% B within 0.01 minutes; Flow Rate: 1.8 ml / minutes; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Temperature: 50 °C; Detection: UV (214 nm, 4 nm; 254 nm, 4 nm) and MS (ESI, Pos mode, 103 to 1000 amu).

[0523] LC-MS Method 8.

[0524]

[0134] Mobile Phase: A: water (10% ammonium bicarbonate) B: acetonitrile, Gradient: 5% B to 95% B within 1.3 minutes, 95% B for 1.7 minutes, back to 5% B within 0.01 minutes; Flow Rate: 1.8 mL / min; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Oven Temperature: 50 °C; Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1000 amu).

[0525] LC-MS Method 9.

[0526]

[0135] Mobile Phase: A: water (0.01% trifluoroacetic acid) B: acetonitrile (0.01% trifluoroacetic acid); Gradient: 5% B to 95% B within 1.3 minutes, 95% B for 1.2 minutes, back to 5% B within 0.5 minutes; Flow Rate: 2.2 mL / min; Column: Chromolith Fast Gradient RP-C18e, 3 x 50 mm; Oven Temperature: 40 °C; Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1300 amu).

[0527] LC-MS Method 10.

[0528]

[0136] Mobile Phase: A: water (0.01% trifluoroacetic acid) B: acetonitrile (0.01% trifluoroacetic acid); Gradient: 5% B to 95% B in 3.0 minutes, 95% B for 2.0 minutes, back to 5% B within 0.01 minutes; Flow Rate: 2.0 mL / min; Column: HALO Cl 8, 4.6 x 30 mm, 2.7 pm; Oven Temperature: 50 °C; Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1200 amu).

[0529] LC-MS Method 11.

[0530]

[0137] Mobile Phase: A: water (0.01% trifluoroacetic acid) B: acetonitrile (0.01% trifluoroacetic acid); Gradient: 10% B to 95% B in 1.4 minutes, 95% B for 1.6 minutes, back to 10% B within 0.01 minutes; Flow Rate: 2.0 mL / min; Column: Sunfire C18, 4.6 x 50 mm, 3.5 pm; Oven Temperature: 50 °C; Detection: UV (214, 4 nm; 254, 4 nm) and MS (ESI, Pos mode, 110 to 1200 amu).

[0531] Preparation of Intermediates

[0532] Intermediate 1. Methyl 5-(benzyloxy)-2-bromobenzimidothioate hydroiodide

[0533] NH S H-l

[0534]

[0535] 46

[0536] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0537]

[0138] To a stirred solution of 5-(benzyloxy)-2-bromobenzothioamide (Intermediate 1A, 22 g, 68.2 mmol) in acetone (300 mL) was added iodomethane (21.2 mL, 341.38 mmol) at room temperature. The reaction mixture was stirred at 50 °C for 5 hours and concentrated to give the title compound (31 g, 98%) as a solid. LC-MS: MS (ESI): 338, 336 m / z [M+H]+, purity: 92% (254 nm). retention time: 1.75 minutes (LC-MS method 1).

[0538] The following intermediate was prepared based on die same procedures as described for intennediate 1.

[0539] Inter.

[0540] No. Structure Name MS m / z [M+H]+methyl 5-(4-amino-2- Br HI NH bromo-6- 373, 375 [M+H]+; RT: 1-1 fluorophenoxy)-2- 1.64 min. (LC-MS H2N '-'' T fluorobenzimidothioate Method 5).

[0541] hydroiodide

[0542]

[0543] Intermediate 1A.5-(Benzyloxy)-2-bromobenzothioamide

[0544]

[0545] H2N

[0546]

[0139] To a stirred solution of 5-benzyloxy-2-bromo-benzonitrile (20 g, 69.4 mmol) in dimethylformamide (200 mL) was added magnesium dichloride hexahydrate (14.1 g, 69.4 mmol) and sodium hydrosulfide hydrate (7.78 g, 138.82 mmol) at room temperature. The reaction was stirred at room temperature for one hour. The reaction mixture was diluted with ethyl acetate (500 mL), washed with water, dried over anhydrous magnesium sulfate, filtered, and concentrated to give the title compound (22 g, 98%) as a yellow solid. LC-MS: MS (ESI): 324, 322 m / z [M+H]+, purity: >99% (254 nm). Retention time: 1.91 minutes (LC-MS method 1).

[0547] The following intennediate was prepared based on the same procedures as described for intennediate 1.

[0548] Inter.

[0549] No. Structure Name MS m / z [M+H]+

[0550]

[0551] 47

[0552] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0553] 5-(4-amino-2-bromo-6- 359, 361 [M+H]+; RT: 1A-1 fluorophenoxy)-2- 1.81 min. (LC-MS fluorobenzothioamide Method 2).

[0554]

[0555] Intermediate 2. (R)-7-((2-((Tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoic acid

[0556] TBSO OH

[0557]

[0558] To a stirred solution of benzyl (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoate (Intermediate 9F, 19.00 g, 28.7 mmol) in methanol (500 mL) was added palladium on carbon (10%, 50% wet, 10 g) and sodium bicarbonate (4.8 g, 57.4 mmol). The mixture was stirred at room temperature under a hydrogen atmosphere for sixteen hours, filtered and concentrated to afford the title compound (16 g. 28.0 mmol, 98%) as a colorless oil. LC-MS: MS (ESI): 593 m / z [M+Na]+, purity: 85% (214 nm). Retention time: 2.22 minutes (LC-MS method 2).

[0559] Intermediate 3. Methyl (R)-3-(3-((R)-l-(2-allylhydrazineyl)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate

[0560]

[0561] A solution of tert-butyl 1 -allyl-2-((R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoyl)hydrazine-l-carboxylate (1.00 eq, 20.00 g, 27.6 mmol) in hydrogen chloride (4M in 1,4-dioxane, 200 mL) was stirred at room temperature for two hours and concentrated. The residue was diluted with saturated sodium bicarbonate (aqueous, 200 mL) and extracted with ethyl acetate (300 mL). The organic layer was washed with water (200 mL x 2), brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by automated flash 48

[0562] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0563] chromatography (120 g silica gel column, eluting with 0-70% ethyl acetate in petroleum ether) to afford the title compound (11.00 g, 78%) as colorless oil. LC-MS: MS (ESI): 511 m / z [M+Na]+, purity: 87% (214 nm). Retention time: 1.54 minutes (LC-MS method 3).

[0564] The following intermediate was prepared based on the same procedures as described for intermediate 3.

[0565] Inter.

[0566] No. Structure Name MS m / z [M+H]+

[0567] 7-((2- hydroxyethyl)sulfonyl)- 379 [M+Na]; RT 1.73 min. (LC-MS 3-1

[0568] 2,6,6-trimethyl-2- method 2) phenylheptanoic acid

[0569]

[0570] Intermediate 3A. Tert-butyl l-allyl-2-((R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3- o

[0571] oxopropyl)phenyl)-2,6,6-trimethylheptanoyl)hydrazine-l-carboxylate

[0572] OOw==

[0573] o

[0574]

[0575] To a stirred solution of (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoic acid (Intermediate 2, 16.00 g, 28.0 mmol) in acetonitrile (200 mL) was added tert-butyl N-allyl-N-amino-carbamate (4827 mg, 30.8 mmol), chloro-N, N, N', N'-tetramethylformamidinium hexafluorophosphate (11.8 g, 42.0 mmol), and 1 -methylimidazole (6903 mg, 98.0 mmol). The mixture was stirred at room temperature for one hour and concentrated. The residue was diluted with ethyl acetate (300 mL), washed with water (200 mL x 2), brine (200 mL), dried over anhydrous sodium sulfate, and concentrated to afford the crude title compound (20 g, 27.6 mmol, 98%) as a white solid. LC-MS: MS (ESI): 747 m / z [M+Na]+, purity: 70% (214 nm). Retention time: 2.38 minutes (LC-MS method 2).

[0576] Intermediate 4.2-Bromo-3-fluoro-6-nitropyridine

[0577]

[0578] 49

[0579] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0580] A mixture of potassium persulfate (5.1 g, 18.8 mmol) in sulfuric acid (5.0 mL, 93.8 mmol) was stirred at 0 °C for 10 minutes, then treated with 6-bromo-5-fluoro-pyridin-2-amine (900 mg, 4.71 mmol) at 0 °C. The mixture was stirred at room temperature for 2 hours, poured into ice water, basified with saturated aqueous NaHCOs to pH ~ 8, and extracted with ethyl acetate (80 mL x 3). The combined organic extracts were washed with brine (80 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (926 mg, 4.19 mmol, 89 %) as a yellow oil. LC-MS: MS (ESI): 221, 223 m / z [M+H]+, purity: 90% (214 nm). Retention time: 1.63 minutes (LC-MS method 2).

[0581] The following intermediate was prepared based on the same procedures as described for intermediate 4.

[0582] Inter.

[0583] No. Structure Name MS m / z [M+H]+

[0584] Br

[0585] 4-bromo-5-fluoro-2- 221, 223 [M+H]; RT 1.63 min. (LC-MS 4-1

[0586] X T nitropyridinc method 4)

[0587] O2N N

[0588] 4-bromo-5-chloro-2- 237, 239 [M+H]; RT 1.82 min. (LC-MS 4-2

[0589] X X nitropyridine method 2) OjN ^ Br

[0590]

[0591] Intermediate 5: Methyl 2-fluoro-5-(methoxymethoxy)benzimidothioate hydroiodide

[0592]

[0593] OMOM

[0594] To a stirred solution of 2-fluoro-5-(methoxymethoxy)benzenecarbothioamide (Intermediate 5B, 1.06 g, 4.92 mmol) in acetone (20 mL) was added methyl iodide (1.5 mL, 24.6 mmol). The mixture was stirred at 50 °C for 3 hours and concentrated to give the title compound (1.70 g, 4.76 mmol, 97 %) as a brown solid. LC-MS: MS (ESI): 230 m / z [M+H]+, purity: 90% (214 nm); Retention time: 1.44 minutes (LC-MS method 2).

[0595] Intermediate 5A: 2-Fluoro-5-(methoxymethoxy)benzonitrile

[0596]

[0597] MOMO

[0598] 50

[0599] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0600] To a stirred solution of 2-fluoro-5-hydroxy -benzonitrile (1.00 g, 7.29 mmol) in dichloromethane (10 mL) was added diisopropylethylamine (3.8 mL, 21.9 mmol) and bromomethyl methyl ether (1.00 g, 8.02 mmol). The mixture was stirred at room temperature for 2 hours, diluted with 80 mL of water, and extracted with ethyl acetate (80 mL x 3). The combined organic phase was washed with brine (80 mL x 2), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by automated silica gel column chromatography (eluting with 0-50% of ethyl acetate in petroleum ether) to give the title compound (1.06 g, 5.85 mmol, 80 %) as a yellow solid. LC-MS: MS (ESI): 182 m / z [M+H]+, purity: 95% (214 nm); Retention time: 1.82 minutes (LC-MS method 2).

[0601] Intermediate 5B: 2-Fluoro-5-(methoxymethoxy)benzothioamide

[0602]

[0603] To a stirred solution of magnesium chloride hexahydrate (1.19 g, 5.85 mmol) and sodium bisulfide (656 mg, 11.7 mmol) in dimethylformamide (20 mL) was added 2-fluoro-5-(methoxymethoxy)benzonitrile (Intermediate 5A, 1.06 g, 5.85 mmol). The mixture was stirred at room temperature for 2 hours, diluted with 100 mL of water, and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with brine (80 mL x 2), dried over anhydrous sodium sulfate, and concentrated to give the title compound (1.06 g, 4.92 mmol, 84 %) as a yellow solid. LC-MS: MS (ESI): 216 m / z [M+H]+, purity: 95% (254 nm); Retention time: 1.66 minutes (LC-MS method 2).

[0604] Intermediate 6: Tert-butyl (4-bromothiazol-2-yl)(tert-butoxycarbonyl)carbamate

[0605] Boc

[0606] J

[0607] Boc7

[0608]

[0609] S

[0610] To a stirred solution of tert-butyl N-(4-bromothiazol-2-yl)carbamate (1.50 g, 5.37 mmol) in toluene (5 mL) were added 4-dimethylaminopyridine (131 mg, 1.07 mmol) and di -tert-butyl dicarbonate (2.7 mL, 11.8 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 2 hours and concentrated. The residue was purified by automated flash chromatography (12 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether)

[0611] 51

[0612] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0613] to give the title compound (2.00 g, 5.27 mmol, 98%) as an oil. LC-MS: MS (ESI): 401, 403 m / z [M+Na]+, purity: 99% (214 nm); Retention time: 2.20 minutes (LC-MS method 2).

[0614] Intermediate 7: Methyl (S)-3-iodo-2-methylpropanoate

[0615] O

[0616]

[0617] To a stirred and cooled (0 °C) solution of methyl (2R)-3-hydroxy-2-methyl-propanoate (100 g, 0.85 mol) in dichloromethane (1.5 L) was added triphenylphosphine (444 g, 1.69 mol), imidazole (115 g, 1.69 mol), followed by addition of iodine (430 g, 1.69 mol) portion wise. The reaction was warmed up to room temperature and stirred overnight. The mixture was filtered. The filtrate was concentrated. The residue was treated with 10% ethyl acetate in petroleum ether (1 L) and then filtered again. The filtrate was concentrated. The residue was purified by automated flash chromatography (330 g silica gel column, eluting with 0-5% ethyl acetate in petroleum ether) to afford the title compound (183 g, 94%) as an oil.1H NMR (400 MHz, CDCl3) δ 3.73 (s, 3H), 3.38 (dd, J= 10.0, 6.4 Hz, 1H), 3.26 (dd, J= 10.0, 6.4 Hz, 1H). 2.83-2.78 (m, 1H), 1.28 (d, J = 6.8 Hz, 3H) ppm.

[0618] The following intermediate was prepared based on the same procedures as described for intermediate 7.

[0619] Inter.

[0620] No. Structure Name MS m / z [M+H]+

[0621] O methyl (R)-3- ¹H NMR (400 MHz, CDCl3) δ 3.72 (s, 3H), 3.39 iodo-2- (dd, J= 10.0, 6.4 Hz, 1H), 3.27 (dd, J= 10.0, 6.4 7-1

[0622] methylpropano Hz, 1H), 2.84-2.78 (m, 1H), 1.28 (d, J= 6.8 Hz, ate 3H) ppm

[0623]

[0624] Intermediate 8: (S)-7-Hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoic acid

[0625]

[0626]

[0140] A chiral salt of enantiomer 1 of (S)-l-(naphthalen-l-yl)ethan-l-aminium 7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 8B, 233 g, 415 mmol) was suspended in ethyl acetate (1000 mL). The mixture was washed with IN HC1 (5 x 500 mL), brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound

[0627] 52

[0628] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0629] (160.00 g, 99 %) as a light-yellow oil. MS (ESI): 413 m / z [M+Na]+, retention time: 1.98 minutes, purity: 89% (214 nm) (LC-MS Method 003).

[0630] Intermediate 8A: 7-((Tert-butyldimethylsilyl)oxy)-2-(3-iodophenyl)-2,6,6-trimethylheptanoic acid

[0631]

[0632]

[0141] To a stirred and cooled (-78 °C) solution of diisopropylamide (1.51 g, 15 mmol) in tetrahydrofuran was added n-butyllithium (2.5M in hexanes, 6 mL, 15 mmol). The reaction was stirred for 30 minutes at this temperature, then hexylmethylphosphoramide was added (2.68 g, 15 mmol) and stirring continued for another 30 minutes. To this mixture was added 2-(3-iodophenyl)propanoic acid (1.66 g, 6 mmol) in 10 mL of tetrahydrofuran over 5 minutes. The resulting mixture was stirred at -78 °C for 30 minutes and tert-butyl((5-iodo-2,2-dimethylpentyl)oxy)dimethylsilane (Intermediate 16-1, 5.33 g, 15 mmol) was added in one portion. The mixture was allowed to warm to room temperature and stirred for 2 hours. After confirming that the starting material was consumed by LC-MS, the reaction was quenched with a mixture of 1 M hydrochloric acid (20 ml) and saturated ammonium chloride solution (15 mL), and extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude title product (1.99 g, 65%). MS (ESI): 505 m / z [M+H]+.

[0633] Intermediate 8B: 7-Hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoic acid

[0634] o

[0635]

[0636]

[0142] To a solution of 7-((tert-butyldimethylsilyl)oxy)-2-(3-iodophenyl)-2,6,6-trimethylheptanoic acid (Intermediate 8A, 4.8 mmol, 2.42 g) in tetrahydrofuran (5 mL) was added 1 M tetrabutylammonium fluoride in THF (9.6 ml, 9.6 mmol). The reaction mixture was stirred at 60 °C for 16 hours. After confirming that the starting material was consumed by LC-MS, the mixture was acidified with 10 mL 1 M hydrochloric acid and extracted with ethyl acetate (2 x 50 ml). The combined organic phases were washed with water, dried over

[0637] 53

[0638] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0639] anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography to afford the title compound (1.03 g, 55%). MS (ESI): 413 m / z [M+Na]+.

[0640] Intermediate 8C: (S)-l-(Naphthalen-l-yl)ethan-l-aminium (R)-7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (-90% ee)

[0641] w

[0642]

[0643]

[0143] To a stirred solution of (S)-(-)-l-(l-naphthyl)ethylamine (127 mL, 705 mmol) in ethyl acetate (7.5 L) was added 7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethyl-heptanoic acid (Intermediate 8B, 500.00 g, 1281 mmol). The reaction was stirred at room temperature overnight. The solid was collected by filtration and the filter cake was washed with ethyl acetate (800 mL). The wet solid was then suspended into EtOAc (1.7 L) and stirred at room temperature for 20 minutes. The mixture was filtered and washed with ethyl acetate (800 mL). The solid was collected and dried in air to give Enantiomer 1 of (S)-l-(naphthalen-l-yl)ethan-l-aminium 7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethyl-heptanoate (288 g, 40 %). Chiral purity: 95% (90% ee). (Chiral conditions: column: AD-H; Mobile phase: n-hexane (0.1% diethylamine): ethanol (0.1% diethylamine) = 95:5; Column temperature: 40 °C; Flow rate: 1.0 mL / minute; Wavelength: 220 nm; Instrument: Shimadzu; Solid salt was de-salted with IN hydrochloric acid in acetonitrile before injection).

[0644] Intermediate 8D: (S)-l-(Naphthalen-l-yl)ethan-l-aminium (R)-7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (-98% ee)

[0645]

[0646]

[0144] A solution of Enantiomer 1 of (S)-l-(naphthalen-l-yl)ethan-l-aminium 7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (-90% ee, Intermediate 8C, 186 g, 315 mmol) in acetonitrile (7.65 L) was refluxed for 5 hours and then stirred at room temperature overnight. The mixture was filtered and the solids rinsed with acetonitrile (2 x 450 mL). The solid was collected and dried in vacuo to give the title compound (153 g, 86 %). Chiral purity: 98% ee, chiral method is the same as in Intermediate 8C).1H NMR (400 MHz, CDCl3) δ 7.84 (t, J= 6.7 Hz, 2H), 7.77 (d, J= 8.2 Hz, 1H), 7.67 (t, J = 1.6 Hz, 1H), 7.56 (d, J = 7.1 Hz, 1H),

[0647] 54

[0648] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0649] 7.52-7.37 (m, 4H), 7.19 (d, 7= 8.5 Hz, 1H), 6.92 (t, 7 = 7.9 Hz, 1H), 4.85 (q, 7 = 6.6 Hz, 1H), 3.19 (dd, 7 = 42.8. 10.4 Hz, 2H), 1.77-1.66 (m, 1H), 1.61-1.57 (m, 1H), 1.54 (d, 7 = 6.7 Hz, 3H), 1.38 (s, 3H), 1.28-0.98 (m, 4H), 0.79 (s, 3H), 0.74 (s, 3H) ppm.

[0650] Intermediate 9: Methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate

[0651]

[0652] o

[0653]

[0145] To a stirred solution of tert-butyl 2-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-2-(3 -((R)-2-methyl-3 -oxopentyl)phenyl)heptanoyl)- 1 -methylhydrazine- 1 -carboxylate (Intermediate 91, 150.00 g, 257 mmol) in dichloromethane (800 mL) was added trifluoroacetic acid (200 mL). The mixture was stirred at room temperature for 4 hours and concentrated. The residue was diluted with water (1000 mL), neutralized with saturated sodium bicarbonate aqueous solution to pH~7, and extracted with ethyl acetate (2 x 800 mL). The combined organic layers were washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (120 g, 97 %) as a light-yellow oil. MS (ESI): 485 m / z [M+H]+; purity: 92% (214 nm); retention time: 1.47 minutes (LC-MS Method 4). ChiraLHPLC purity: 99.4% (>99% ee, 254 nm), RT = 1.75 minutes (chiral SFC column conditions: Column: AD-3, 4.6 x 100 mm, 3 pm; mobile phase: carbon dioxide / methanol (0.2% 7M ammonia in methanol) = 80:20); Injection volume: 5.00 uL; Run time: 6.0 minutes; Flow rate: 3.0 mL / minute; Wavelength: 214 nm; Column temperature: 40 °C).

[0654]

[0146] The following intermediates were prepared based on the same procedures as described for intermediate 9.

[0655] Inter. MS m / z No. Structure Name [M+H]+N'-(2-(benzyloxy)ethyl)-7-((2- 743 [M+H]; ((tert- RT 2.35 min.

[0656] 9-1TBDPSO^SOJ<^NN^OBn butyldiphenylsilyl)oxy)ethyl)sulf

[0657] ° O H (LC-MS onyl)-2,6,6-trimethyl-2- method 2) phenylheptanehydrazide

[0658]

[0659] 55

[0660] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0661] methyl (S)-3-(3-((R)-7-((2- hydroxyethyl)sulfonyl)-2,6,6- 485 [M+H];

[0662] trimethyl-l-(2- RT 1.47 min.

[0663] 9-2

[0664] methylhydrazineyl)- 1 - (LC-MS oxoheptan-2-yl)phenyl)-2- method 4) methylpropanoate

[0665] 0 421 [M+H];

[0666] 2-(3,5-difluorophenyl)-7-((2- RT 1.61 min.

[0667] 9-3 H A o' OH hydroxyethyl)sulfonyl)-N’, 2,6,6- (LC-MS tetramethylheptanehydrazide

[0668] method 2) O'' methyl (R)-3-(3-((R)-l-(2- 511 [M+H]; allylhydrazineyl)-7-((2- RT 1.54 min.

[0669] 9-4 ^~NH OH hydroxyethyl)sulfonyl)-2,6,6- (LC-MS trimethyl- 1 -oxoheptan-2- method 3) A A yl)phenyl)-2-methylpropanoate

[0670]

[0671] Intermediate 9A: Benzyl (R)-7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate

[0672] 0

[0673]

[0674]

[0147] To a stirred solution of (R)-7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoic acid (Intermediate 8, 160.00 g, 410 mmol) in N, N-dimethylformamide (800 mL) was added benzyl bromide (73.6 g, 430 mmol) followed by potassium carbonate (85.00 g, 615 mmol). The reaction was stirred at room temperature for 16 hours, then diluted with water (1.5 L), and extracted with ethyl acetate (3 x 800 mL). The combined organic extracts were washed with brine (3 x 1.0 L), dried over anhydrous sodium sulfate and concentrated. The residue was purified by automated flash chromatography (330 g silica gel column, eluting with 0-25% ethyl acetate in petroleum ether) to give the title compound (190.00 g, 96 %) as a light-yellow oil. ’H NMR (400 MHz, CDCh) 8 7.62 (t, J = 1.7 Hz, 1H), 7.57 (d, J = 7.8 Hz, 1H), 7.36 - 7.28 (m, 3H), 7.25 - 7.19 (m, 3H), 7.02 (t, 7= 7.9 Hz, 1H), 5.11 (dd, 7= 29.3, 12.4 Hz, 2H), 3.23 (d, 7= 4.4 Hz, 2H), 2.04 - 1.97 (m, 1H), 1.86 - 1.75 (m, 1H), 1.62 (s, 1H), 1.53 (s, 3H), 1.30 - 1.15 (m, 4H), 0.79 (d, 7= 1.9 Hz, 6H). MS (ESI): 503 m / z [M+Na]+; Purity: 98% (214 nm); Retention time: 2.14 minutes (LC-MS Method 4). Chiral-HPLC: RT = 1.630 min, 99% (98% ee) (chiral SFC column conditions: Column: OJ-3, 4.6 x 100 mm, 3 pm; mobile phase: carbon dioxide / methanol (0.2% 7M ammonia in methanol) (90:10); injection volume: 3.00 uL; Run

[0675] 56

[0676] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0677] time: 6.0 minutes; Flow rate: 3.0 mL / minute; Back-pressure: 2000 psi; Column temperature: 40 °C.)

[0678]

[0148] The following intermediates were prepared based on the same procedures as described for intermediate 9A.

[0679] Inter. MS m / z No. Structure Name [M+H]+0 481 [M+H];

[0680] benzyl 7-hydroxy-2-(3- B H RT 2.39 min.

[0681] 9A-1 iodophenyl)-2,6,6- (LC-MS Cl trimethylheptanoate

[0682] method 1)

[0683]

[0684] Intermediate 9B: Benzyl (R)-7-(acetylthio)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate o

[0685]

[0686]

[0149] To a stirred and cooled (0 °C) solution of triphenylphosphine (207.48 g, 791 mmol) in tetrahydrofuran (1.5 L) was added diisopropyl azodicarboxylate (154 mL, 791 mmol) dropwise at 0 °C. The mixture was stirred at 0 °C until the formation of a white solid was observed. A solution of benzyl (R)-7-hydroxy-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 9A, 190.00 g, 396 mmol) and thioacetic acid (2.00 eq, 57 mL, 791 mmol) in tetrahydrofuran (200 mL) was added dropwise at 0 °C. After the addition, the mixture was stirred at 0 °C for 1 hour, then at room temperature for 16 hours and concentrated. The residue was slurried with 10% ethyl acetate in petroleum ether (1.0 L) for 0.5 hours. The mixture was filtered, and the filtrate was concentrated. The residue was purified by automated flash chromatography (two 330 g silica gel columns, eluting with 0-10% ethyl acetate in petroleum) to give the title compound (200.00 g, 94 %) as light-yellow oil. MS (ESI): 561 m / z [M+Na]+; purity: 91% (214 nm); Purity: 91% (214 nm); retention time: 2.39 minutes (LC-MS Method 4). Chiral-HPLC purity: RT = 1.35 min, 99.3% (98.6% ee) (chiral SFC column conditions: Column: OJ-3, 4.6 x 100 mm, 3 pm; mobile phase: carbon dioxide / methanol (0.2% 7M ammonia in methanol) (90:10); injection volume: 0.80 uL; Run time: 4.0 minutes; Flow rate: 3.0 mL / minute; Backpressure: 2000 psi; Column temperature: 40 °C.)

[0687]

[0150] The following intermediates were prepared based on the same procedures as described for intermediate 9B.

[0688] 57

[0689] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0690] Inter. MS m / z No. Structure Name [M+H]+

[0691] 10437 [M+NaJ;

[0692] tert-butyl 7-(acetylthio)-2-(3,5- RT 2.49 min.

[0693] 9B-1 difluorophenyl)-2,6,6- (LC-MS A trimethylheptanoate

[0694] method 2) 0 539 [M+H];

[0695] benzyl 7-(acetylthio)-2-(3- B S Ac RT 2.49 min.

[0696] 9B-2 iodophenyl)-2,6,6- (LC-MS UL trimethylheptanoate

[0697] method 8)

[0698]

[0699] Intermediate 9C: Benzyl (R)-7-((2-hydroxyethyl)thio)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate

[0700] o

[0701]

[0702]

[0151] To a stirred and cooled (0 °C) solution of benzyl (R)-7-(acetylthio)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 9B, 200.00 g, 390 mmol) in ethanol (500 mL) was added 2-bromoethanol (34 mL, 483 mmol) and sodium ethoxide (20% in ethanol, 189.56 g, 557 mmol) dropwise. The mixture was stirred at 0 °C for 1.0 hour, quenched with water (1.0 L), and extracted with ethyl acetate (3 x 600 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by automated flash chromatography (330 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (190 g, 95 %) as a light-yellow oil. MS (ESI): 563 m / z [M+Na]+; purity: 86% (214 nm); (214 nm); retention time: 2.28 minutes (LC-MS Method 4). Chiral-HPLC purity: RT = 2.10 min, 99.5% (99% ee) (chiral SFC column conditions: Column: OJ-3, 4.6 x 100 mm, 3 pm; mobile phase: carbon dioxide / methanol (0.2% 7M ammonia in methanol) (90:10); injection volume: 5.00 uL; Run time: 6.0 minutes; Flow rate: 3.0 mL / minute; Back-pressure: 2000 psi; Column temperature: 40 °C.)

[0703]

[0152] The following intermediates were prepared based on the same procedures as described for intermediate 9C.

[0704] Inter. MS m / z No. Structure Name [M+H]+

[0705]

[0706] 58

[0707] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0708] 439 [M+Na]; A 1

[0709] / '.'O' A-0

[0710] ''g'',o tert-butyl 2-(3, 5 -difluorophenyl) - RT 2.28 min.

[0711] 9C-1 A 7-((2-hydroxyethyl)thio)-2,6,6- (LC-MS trimethylheptanoate

[0712] method 2) o 563 [M+Na];Bbenzyl 7-((2-hydroxyethyl)thio)- nO-Sp^-S~°HRT 1.57 min.

[0713] 9C-2 2-(3-iodophenyl)-2,6,6- (LC-MS CL trimethylheptanoate

[0714] method 9)

[0715]

[0716] Intermediate 9D: Benzyl (R)-7-((2-hydroxyethyl)sulfonyl)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate

[0717] o

[0718]

[0719]

[0153] To a stirred and cooled (0 °C) solution of benzyl (R)-7-((2-hydroxyethyl)thio)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 9C, 190 g, 352 mmol) in methanol (800 mL) was added dropwise a solution of ammonium molybdate tetrahydrate (65.00 g, 52.6 mmol) in hydrogen peroxide (30% in water, 250 mL). The mixture was stirred at room temperature for 2 hours, then diluted with water (1200 mL) and extracted with ethyl acetate (3 x 500 mL). The combined organic extracts were washed with brine (5 x 500 mL), dried over sodium sulfate, and concentrated to give the title compound (190 g, 94 %) as a colorless oil. MS (ESI): 595 m / z [M+Na]+; purity: >99% (214 nm); retention time: 2.00 minutes (LC-MS Method 4). Chiral-HPLC purity: RT = 10.24 min, 99.0% (98% ee) (chiral column conditions: Column: AY-H, 4.6 x 250 mm, 5 pm; mobile phase: Hexanes (0.1% diethylamine):ethanol (0.1% diethylamine) = 80:20); Injection volume: 10.00 uL; Run time: 30.0 minutes; Flow rate: 1.0 mL / minute; Instrument: Shimadzu; Wavelength: 214 nm and 254 nm.)

[0720]

[0154] The following intermediate was prepared based on the same procedures as described for intermediate 9D.

[0721] Inter. MS m / z No. Structure Name [M+H]+o I 471 [M+Na]; o | ij A tert-butyl 2-(3, 5 -difluorophenyl) - RT 2.09 min.

[0722] 9D-1 A 7-((2-hydroxyethyl)sulfonyl)- (LC-MS 2, 6, 6-trimethy lheptanoate

[0723] method 2)

[0724]

[0725] 59

[0726] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0727] benzyl 7-((2- 595 [M+Na]; hydroxyethyl)sulfonyl)-2-(3- RT 1.45 min.

[0728] 9D-2

[0729] iodophenyl)-2,6,6- (LC-MS Cl

[0730] trimethylheptanoate method 9)

[0731] 435 [M+Na]; 1 / u 0 tert-butyl 7-((2- RT 1.73 min.

[0732] 9D-3 ' 6 hydroxyethyl)sulfonyl)-2,6,6- (LC-MS trimethyl-2-phenylheptanoate

[0733] method 2)

[0734]

[0735] Intermediate 9E: Benzyl (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate

[0736] o

[0737]

[0738]

[0155] To a stirred solution of benzyl (R)-7-((2-hydroxyethyl)sulfonyl)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 9D, 190 g, 332 mmol) in dichloromethane (1.0 L) was added tert-butyldimethylsilyl chloride (52.52 g, 348 mmol) and imidazole (33.89 g, 498 mmol). The mixture was stirred at room temperature for 16 hours and concentrated. The residue was purified by automated flash chromatography (330 g x 2 silica gel column eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (210.00 g, 92 %) as a colorless oil. ’H NMR (400 MHz, CDCk) 5 7.54 (s, 1H), 7.48 (d, J= 7.8 Hz, 1H), 7.28-7.12 (m, 6H), 6.93 (t, J= 7.9 Hz, 1H), 5.06-4.94 (m, 2H), 3.99-3.90 (m, 2H), 3.00-2.86 (m, 4H), 1.98-1.90 (m, 1H), 1.83-1.70 (m, 1H), 1.50-1.34 (m, 5H), 1.10-0.95 (m. 8H), 0.81 (s, 9H), 0.00 (s, 6H) ppm. MS (ESI): 709 m / z [M+Na]+; purity: 98% (214 nm); retention time: 2.48 minutes (LC-MS Method 4). Chiral-HPLC purity: RT = 29.43 min, >99% (>99% ee) (chiral column conditions: Column: IC, 4.6 x 250 mm, 5 pm; mobile phase: Hexanes (0.1% diethylamine): isopropanol (0.1% diethylamine) = 90:10); Injection volume: 10.00 uL; Run time: 45.0 minutes; Flow rate: 1.0 mL / minute; Instrument: Shimadzu; Wavelength: 214 nm; Column temperature: 40 °C.).

[0739]

[0156] The following intermediates were prepared based on the same procedures as described for intermediate 9E.

[0740] Inter. MS m / z No. Structure Name [M+H]+

[0741]

[0742] 60

[0743] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0744] benzyl 7-((2-(tert- 817 [M+Na]; U o

[0745] butyldiphenylsilyl)ethyl)sulfonyl RT 2.85 min.

[0746] 9E-1 / \ II

[0747] A. 'x0

[0748] )-2-(3-iodophenyl)-2,6,6- (LC-MS L l

[0749] trimethylheptanoate method 5) 7-((2-((tert- 617 [M+Na]; H,QU O

[0750] butyldiphenylsilyl)oxy)ethyl)sulf RT 3.66 min.

[0751] 9E-2 -X ' 0 onyl)-2,6,6-trimethyl-2- (LC-MS phenylheptanoic acid method 10)

[0752]

[0753] Intermediate 9F: Benzyl (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoate

[0754] o

[0755] BnO

[0756] OTBS

[0757]

[0758] O

[0759]

[0157] To a stirred suspension of zinc (119.95 g, 1835 mmol) in N, N-dimethylformamide (800 mL) was added iodine (7.76 g, 30.6 mmol) under argon. The reaction mixture was stirred at room temperature for 40 minutes, then treated with methyl (2S)-3-iodo-2-methyl-propanoate (Intermediate 7, 104.59 g, 459 mmol), and stirred for another 40 minutes. To this mixture was added 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (12.55 g, 30.6 mmol), tris(dibenzylidene-acetone)-dipalladium (14.00 g, 15.3 mmol), benzyl (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-iodophenyl)-2,6,6-trimethylheptanoate (Intermediate 9E, 210.00 g, 306 mmol). The reaction mixture was stirred at room temperature for 16 hours, diluted with water (2000 mL) and ethyl acetate (1500 mL). The solid was filtered off. The filtrate phases were separated. The aqueous phase was extracted with ethyl acetate (2 x 1000 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by automated flash chromatography (2 x 330 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (186 g, 92 %) as light-yellow oil. MS (ESI): 683 mz [M+Na]+; purity: 89% (214 nm); retention time: 2.35 minutes (LC-MS Method 4). Chiral-HPLC purity: >99% (>99% ee, 254 nm), RT = 19.39 min, (chiral column conditions: Column: AY-H, 4.6 x 250 mm, 5 pm; mobile phase: Hexanes (0.1% diethylamine): ethanol (0.1% diethylamine) = 95:5); Injection volume: 10.00 uL; Run time: 30.0 minutes; Flow rate: 1.0 mL / minute; Instrument: Shimadzu; Wavelength: 214 nm and 254 nm).

[0760] 61

[0761] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0762]

[0158] The following intermediates were prepared based on the same procedures as described for intermediate 9F.

[0763] Inter. MS m / z No. Structure Name [M+H]+benzyl (R)-7-((2-((tert- 807 m / z butyldiphenylsilyl)oxy)ethyl)sulf

[0764] LM+Na] RT 9F-1 onyl)-2-(3-((S)-3-methoxy-2- UUYK 2.54 min. (LC- methyl-3 -oxopropyljphenyl)- MS method 4) 2,6, 6-trimethy lheptanoate

[0765] benzyl (R)-7-((2-((tert- 683 m / z butyldimethylsilyl)oxy)ethyl)sulf ^pC^ ^ [M+Na]; RT 9F-2 onyl)-2-(3-((S)-3-methoxy-2- UUy 2.43 min. (LC- methyl-3-oxopropyl)phenyl)- MS method 4) 2, 6, 6-trimethy lheptanoate

[0766]

[0767] Intermediate 9G: Benzyl (R)-7-((2-hydroxyethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoate

[0768] o

[0769]

[0770] o

[0771]

[0159] To a solution of benzyl (R)-7-((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoate (Intermediate 9F, 186.00 g, 281 mmol) in tetrahydrofuran (200 mL) was added tetra-n-butylammonium fluoride (IN in tetrahydrofuran, 422 mL, 422 mmol). The reaction mixture was stirred at room temperature for 2 hours, then diluted with ethyl acetate (1000 mL). The solution was washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by automated flash chromatography (330 g silica gel column, eluting with 0-85% ethyl acetate in petroleum) to give the title compound (145 g, 94 %) as a light-yellow oil. MS (ESI): 569 m / z [M+Na]+; purity: 97% (214 nm); retention time: 1.92 minutes (LC-MS Method 4). Chiral-HPLC purity: 99.4% (>99% ee, 214 nm), RT = 11.99 minutes (chiral column conditions: Column: OJ-H, 4.6 x 250 mm, 5 pm; mobile phase: Hexanes (0.1% diethylamine): ethanol (0.1% diethylamine) = 80:20); Injection volume: 10.00 uL; Run time: 30.0 minutes; Flow rate: 1.0 mL / minute; Instrument: Shimadzu; Wavelength: 214 nm and 254 nm).

[0772] 62

[0773] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0774]

[0160] The following intermediate was prepared based on the same procedures as described for intermediate 9G.

[0775] Inter. MS m / z No. Structure Name [M+H]+benzyl (R)-7-((2- 569 [M+Na] hydroxyethyl)sulfonyl)-2-(3- nr T A ^ RT 1.97 min.

[0776] 9G-1 ((S)-3-methoxy-2-methyl-3- (LC-MS oxopropyljpheny 1) -2, 6,6- method 4) trimethylheptanoate

[0777]

[0778] Intermediate 9H: (R)-7-((2-Hydroxyethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoic acid

[0779]

[0780]

[0161] To a stirred solution of benzyl (R)-7-((2-hydroxyethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoate (Intermediate 9G, 145 g, 265 mmol) in methanol (1000 mL) was added 10% palladium on carbon (wetted with ca. 55% Water, 20 g). The reaction mixture was stirred under hydrogen for 6 hours at 50 °C, cooled to room temperature and filtered through a pad of Celite. The filtrate was concentrated to give the title compound (120 g, 99 %) as a colorless oil. MS (ESI): 479 m / z [M+Na]+; purity: >99% (214 nm); retention time: 1.66 minutes (LC-MS Method 4). Chiral-HPLC purity: 99.5% (99% ee, 214 nm), RT = 1.61 minutes (chiral SFC column conditions: Column: IG-3, 4.6 x 100 mm, 3 pm; mobile phase: carbon dioxide / methanol (0.2% 7M ammonia in methanol) = 65:35); Injection volume: 5.00 uL; Run time: 6.0 minutes; Flow rate: 3.0 mE / minute; Back pressure: 2000 psi; Wavelength: 214 nm; Column temperature: 40 °C).

[0781]

[0162] The following intermediates were prepared based on the same procedures as described for intermediate 9H.

[0782] Inter. MS m / z No. Structure Name [M+H]+P (R)-7-((2-((tert- 717.0 [M+Na]; 9H-1 butyldiphenylsilyl)oxy)ethyl)sulf RT 2.28 min.

[0783] onyl)-2-(3-((S)-3-methoxy-2- (LC-MS methyl-3-oxopropyl)phenyl)- method 4)

[0784]

[0785] 63

[0786] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0787] 2,6,6-trimethylheptanoic acid

[0788] (R)-7-((2-((tert- 593.0 [M+Na]; butyldimethylsilyl)oxy)ethyl)sulf

[0789] RT 2.22min.

[0790] 9H-2 onyl)-2-(3-((R)-3-methoxy-2- (LC-MS methyl-3 -oxopropyl)phenyl)- method 2) 2,6,6-trimethylheptanoic acid

[0791]

[0792] Intermediate 91: Tert-butyl 2-((R)-7-((2-hydroxyethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoyl)-l-methylhydrazine-l-carboxylate

[0793] Boc O

[0794] 0

[0795] o

[0796] 07 / ==

[0797]

[0163] To a stirred solution of (R)-7-((2-Hydroxyethyl)sulfonyl)-2-(3-((R)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoic acid (Intermediate 9H, 120 g, 263 r / . / c

[0798] mmol) in acetonitrile (8 / 0°=0 mL) was added tert-butyl N-amino-N-methyl-carbamate (40.34 g,

[0799] \, xO \ °

[0800] 276 mmol), N, N, N xZ<) '

[0801] O’', N’ -tetramethylchloroformamidinium-hexafluorophosphate (73.72 g, 263 mmol) and 1-methylimidazole (73 mL, 920 mmol). The reaction mixture was stirring at room co

[0802] temperature 00

[0803] 1- for 1 hour, then diluted with ethyl acetate (1500 mL). The solution was washed with 0.5 N hydrochloric acid, water, saturated sodium bicarbonate solution, brine, dried over sodium sulfate, and concentrated to give the title compound (150 g, 98 %) as a light -yellow oil. MS (ESI): 607 m / z [M+Na]+; purity: 86% (214 nm); retention time: 1.76 minutes (LC-MS Method 4). Chiral-HPLC purity: 99.6% (>99% ee, 254 nm), RT = 15.39 minutes (chiral column conditions: Column: AD-H, 4.6 x 250 mm, 5 pm; mobile phase: hexanes (0.1 % diethylamine) / ethanol (0.1% diethylamine = 90:10); Injection volume: 10.00 uL; Run time: 30.0 minutes; Flow rate: 1.0 mL / minute; Wavelength: 214 nm and 254 nm; Column temperature: 40 °C).

[0804]

[0164] The following intermediate was prepared based on the same procedures as described for intermediate 91.

[0805] Inter.

[0806] MS m / z [M+H] No. Structure Name+tert-butyl 2-(2-(3,5- 543 [M+Na] 91-1 difluorophenyl)-7 -((2- RT: 1.84 min.

[0807] hydroxyethyl)sulfonyl)-2,6,6- (LC-MS

[0808]

[0809] 64

[0810] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0811] trimethy lheptanoy 1) - 1 - method 5) methylhydrazine- 1 -carboxylate

[0812] tert-butyl l-(2- (benzyloxy)ethyl)-2-(7-((2- 865 [M+Na] ((tert- RT: 3.42 min.

[0813] 91-2 butyldiphenylsilyl)oxy)ethyl)sul

[0814] O Boc (LC-MS fony 1) -2, 6, 6-trimethy 1-2- method 4) phenylheptanoyl)hydrazine- 1 - carboxylate

[0815] tert-butyl l-allyl-2-((R)-7-((2- ((tert- / / _? butyldimethylsilyl)oxy)ethyl)su 747 [M+Na] / 0 J i. OTBS lfonyl)-2-(3-((R)-3-methoxy-2- RT: 2.38 min.

[0816] 91-3 BocN ’ <r

[0817] methy 1-3 -oxopropyl)phenyl) - (LC-MS 2,6,6- method 2) trimethylheptanoyl)hydrazine- 1 -carboxylate

[0818]

[0819] Intermediate 10: Methyl (S)-3-(3-((R)-2-(5-(2-fluoro-5-hydroxyphenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate

[0820]

[0821]

[0165] To a stirred solution of methyl (S)-3-(3-((R)-2-(5-(5-(benzyloxy)-2-tluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate (3.60 g, 5.19 mmol) in ethanol (60 mL) was added palladium on carbon (10%, 3.63 g, 3.41 mmol). The mixture was stirred at 50 °C for 24 hours under a hydrogen atmosphere. The mixture was filtered and concentrated. The residue was purified by automated flash chromatography (20 g silica gel column, 0-50% ethyl acetate in petroleum ether) to give the title compound (3.00 g,4.47 mmol, 86 %) as a white solid. MS (ESI): 604 m / z [M+H]+; purity: 95% (254 nm); retention time: 1.86 minutes (LC-MS Method 2).

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[0823] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0824] Intermediate 10A: Methyl (S)-3-(3-((R)-2-(5-(5-(benzyloxy)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate

[0825]

[0826]

[0166] To a stirred solution of methyl 5-(benzyloxy)-2-fluorobenzimidothioate hydroiodide (Intermediate 1, 3.40 g, 8.44 mmol)and methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate hydrochloride (4.00 g, 7.68 mmol) in pyridine (12 mL) was added 4A MS (3.5 g) at room temperature. The reaction mixture was stirred at 50 °C overnight. The reaction mixture was filtered. The filtrate was concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-100% ethyl acetate in petroleum ether) to give the title compound (3.60 g,5.19 mmol, 68 %) as a white solid. MS (ESI): 694 mJz [M+H]+; purity: 95% (254 nm); retention time: 2.09 minutes (LC-MS Method 3).

[0827] Intermediate 11: (3-(Bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide

[0828]

[0829] NH OAc Br

[0830]

[0167] To a stirred solution of (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(3-carbamothioyl-4-fluorophenyl)methyl acetate (0.80 g, 1.17 mmol) in acetone (20 mL) was added methyl iodide (0.37 mL, 5.84 mmol) at room temperature. The reaction mixture was stirred at 50 °C for 6 hours and concentrated to give the title compound (0.95 g, 98 %) as an orange solid. MS (ESI): 697, 699 mJz [M+H]+; purity: 90% (214 nm); retention time: 2.48 minutes (LC-MS Method 4).

[0831]

[0168] The following intermediates were prepared based on the same procedures as described for intermediate 11.

[0832] Inter.

[0833] No. Structure Name MS mJz [M+H]+

[0834]

[0835] 66

[0836] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0837] (3-(bis(tert- N(BOC)2butoxycarbonyl)amino)-6-bromo- 681, 683 [M+H]+;

[0838] 5 -chloro-2,4-difluorophenyl) ( 4- RT: 2.04 min.

[0839] 11-1

[0840] fluoro-3- (LC-MS Method Hl NH OAc Br (imino(methylthio)methyl)phenyl 5).

[0841] )methyl acetate hydroiodide

[0842] methyl 5-((2-(bis(tert- HI NH 578, 580 [M+H]+;

[0843] butoxycarbony 1) amino) -4- BOC q C _ JI / RT: 2.14 min.

[0844] 11-2 bromothiazol-5 -yl)thio)-2-Boc;n^ N X'T Txs(LC-MS Method BrFfluorobenzimidothioate

[0845] 2). hydroiodide

[0846] Br HI NH methyl 5-(4-amino-2-bromo-6- 373, 375 [M+H]+;

[0847] fluorophenoxy)-2- RT: 1.64 min.

[0848] 11-3

[0849] fluorobenzimidothioate (LC-MS Method H2N TX^ / ^FxT hydroiodide 5).

[0850] (4-(bis(tert- 629, 631 [M+H]+: Br OAc NH HI butoxycarbonyl)amino)-2-bromo- RT: 1.93 min.

[0851] 11-4 6-fluorophenyl)(4-fluoro-3- (LC-MS Method (Boc^N A^^AF A (imino(methylthio)methyl)phenyl

[0852] 5).

[0853] )methyl acetate hydroiodide

[0854] (3-(bis(tert- NH OAc F butoxycarbonyl)amino)-6-bromo- 697, 699 [M+H]+;

[0855] N<B°C>2 4,5-dichloro-2-fluorophenyl)(4- RT: 2.48 min.

[0856] 11-5HIY T

[0857] F'VBr^ I' yT' OI fluoro-3- (LC-MS Method Cl (imino(methylthio)methyl)phenyl 4).

[0858] )methyl acetate hydroiodide

[0859] methyl 5-(4-amino-6-bromo-2,3- 391, 393 [M+H]+; F HI NH

[0860] difluorophenoxy)-2- RT: 1.90 min.

[0861] 11-6 T T I T fluorobenzimidothioate (LC-MS Method H2N'xX^'BrX-'' T

[0862] hydroiodide 4). methyl 5-(4-(bis(tert- 591, 593 [M+H]+; F HI NH butoxycarbonyl)amino)-6-bromo- RT: 2.47 min.

[0863] 11-7 A A 2,3-difluorophenoxy)-2- (LC-MS Method (Boc)2Nx^- / xBr fluorobenzimidothioate

[0864] 2). hydroiodide

[0865] F OAc i ll (4-(bis(tert- 665. 667 [M+H]+:

[0866] RT: 2.47 min.

[0867] 11-8 butoxycarbonyl)amino)-2-bromo- Boc2NFX'AX / k~BrTNH3,5,6-trifluorophenyl)(4-fluoro-3- (LC-MS Method F

[0868] (imino(methylthio)methyl)phenyl 2).

[0869]

[0870] 67

[0871] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0872] )methyl acetate hydroiodide

[0873] methyl 5-((3-(bis(tert- NBOC2butoxycarbonyl)amino)-6-bromo- 623, 625 [M+H]+:

[0874] 2,4,5- RT: 2.18 min.

[0875] 11-9

[0876] trifluorophenyl)(hydroxy)methyl) (LC-MS Method Br OH NH -2-fluorobenzimidothioate 3).

[0877] hydroiodide

[0878] NH HI F methyl 5-(3-amino-6-bromo- 407, 409 [M+H]+;

[0879] 2,4,5-trifluorobenzyl)-2- RT: 1.88 min.

[0880] 11-10

[0881] ^sYndxNH2fluorobenzimidothioate (LC-MS Method F hydroiodide 7).

[0882] F methyl 5-(3-amino-2-bromo- 409, 411 [M+H]+;

[0883] 4,5,6-trifluorophenoxy)-2- RT: 2.02 min.

[0884] 11-11

[0885] k yXX oXX NH2fluorobenzimidothioate (LC-MS Method NH HI Br hydroiodide 4).

[0886] (4-(bis(tert- NH HI OAc Br 647, 649 [M+H]+;

[0887] butoxycarbonyl)amino)-2-bromo- RT: 2.23 min.

[0888] 11-12 3,6-difluorophenyl)(4-fluoro-3- (LC-MS Method F'^'^NBOC, (imino(methylthio)methyl)phenyl

[0889] 4).

[0890] )methyl acetate hydroiodide

[0891] HI F methyl 5-((4-amino-6-bromo-2,3- 407, 409 [M+H]+;

[0892] difluorophenyl)thio)-2- RT: 2.23 min.

[0893] 11-13

[0894] HNYrsAFfluorobenzimidothioate (LC-MS Method F '-^ Br'^ / XNH2hydroiodide 2).

[0895] (4-(bis(tert- butoxycarbonyl)amino)-6-bromo- 2,3-difluorophenyl)(4-fluoro-3- NHulOAc F (imino(methylthio)methyl)phenyl 647, 649 [M+H]+;

[0896] )methyl acetate hydroiodide RT: 2.29 min. '■;w irL. (LC-MS Method 11-14 and

[0897] NH, „ OAc F 4) and 547, 549

[0898] (6-bromo-4-((tert- [M+H]+; (LC-MS Tox butoxycarbonyl)amino)-2,3- Method 4) difluorophenyl)(4-fluoro-3- (imino(methylthio)methyl)phenyl

[0899] )methyl acetate hydroiodide

[0900] NHH| Br

[0901] methyl 5-(4-amino-2-bromo-3- 407, 409 [M+H]+; 11-15

[0902] Xjf YT chloro-6-fluorophenoxy)-2- RT: 1.69 min. F^-"^ F^^^NHj fluorobenzimidothioate (LC-MS Method

[0903]

[0904] 68

[0905] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0906] hydroiodide 5).

[0907] NH HI Br methyl 5-(4-amino-2-bromo-3,6- 391, 393 [M+H]+;

[0908] difluorophenoxy)-2- RT: 1.97 min. 11-16

[0909] fluorobenzimidothioate (LC-MS Method F'^^'NH2

[0910] hydroiodide 3). methyl 5-((2-bromo-6-fluoro-4- NH HI Br 524, 526 [M+H]+;

[0911] ((4-methoxybenzyl)oxy)-3- RT: 2.01 min. 11-17 methylphenyl)thio)-2- (LC-MS Method F'"^-" X)PMB fluorobenzimidothioate

[0912] 2). hydroiodide

[0913] S''' Br methyl 5-(2-bromo-3,6-difluoro- 406, 408 [M+H]+;

[0914] 4-methoxyphenoxy)-2- RT: 1.68 min. 11-18

[0915] HNACf°liFfluorobenzimidothioate (LC-MS Method hydroiodide 2).

[0916]

[0917] Intermediate HA: tert-Butyl (4-bromo-2,3-dichloro-6-fluorophenyl)(tert-butoxycarbonyl)carbamate

[0918]

[0919]

[0169] To a stirred solution of 4-bromo-2,3-dichloro-6-fluoro-aniline (2.89 g, 11.2 mmol), N, N-diisopropylethylamine (7.8 mL, 44.7 mmol) and 4-dimethylaminopyridine (545 mg, 4.47 mmol) in tetrahydrofuran (45 mL) was added di-tert-butyl dicarbonate (7.8 mL, 33.5 mmol). The mixture was stirred at room temperature for 16 hours. The mixture was diluted with water (120 mL), extracted with ethyl acetate (90 mL x 2). The combined organic extracts were washed with water (30 mL), brine (30 mL), dried over anhydrous sodium sulphate, filtered, and concentrated. The residue was purified by column chromatography on silica (80 g silica gel column; eluting with ethyl acetate in petroleum ether from 0 to 8%) to give the title compound (3.80 g, 74 %) as a solid. MS (ESI): 480, 482 m / z [M+Na]+; purity: >99% (254 nm); retention time: 2.33 minutes (LC-MS Method 2).

[0920]

[0170] The following intermediates were prepared based on the same procedures as described for intermediate HA.

[0921] Structure Name MS m / z [M+H]+

[0922]

[0923] Inter.

[0924] 69

[0925] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0926] No.

[0927] tert-butyl (4-bromo-3- ’H NMR (400 MHz, F

[0928] chloro-2,6- CDCb) 5 7.30 (dd, J = (Boc^N^ J^ / CI

[0929] 11A-1 T I difluorophenyl) (tert- 8.8, 2.4 Hz, 1H), 1.44 (s, butoxycarbonyl)carba

[0930] 18H) ppm.

[0931] mate

[0932] 'H NMR (500 MHz, DMSO- Br tert-butyl (3-bromo-5- d6) 57.65 - 7.50 (dt, 7 = 8.5, fluorophenyl) (tert- 11A-2 2.0 Hz„ 1H), 7.40 (s, 1H), butoxycarbonyl)carba

[0933] (Boc)2N XI F 7.30 (dt.7= 9.5, 2.0 Hz, 1H), mate

[0934] 1.39 (s, 18H)ppm tert-butyl (4-bromo- Cl

[0935] 2,3-dichloro-6- (BOC)2N' 4\. CI 480, 482 [M+Na]+; RT: 2.33 11A-3 XI fluorophenyl) (tert- min. (LC-MS Method 2). F^-^ Br butoxycarbonyl)carba

[0936] mate

[0937] tert-butyl (5-bromo-4- (3-cyano-4- fluorophenoxy ) -2,3- 543, 545 [M+H]+; RT: 2.33 11A-4 I T I T difluorophenyl) (tert- min. (LC-MS Method 2).

[0938] butoxycarbonyl)carba

[0939] mate

[0940] tert-butyl (3-bromo- NBOC2

[0941] F2,5,6- ^ / L,F426, 428 [M+H]+; RT: 2.71 11A-5 J T trifluorophenyl)(tert- min. (LC-MS Method 4). Br / ^'"'' T butoxycarbonyl)carba

[0942] mate

[0943] F di-tert-butyl (4-bromo-1H NMR (400 MHz, CDCh) BOC2NF2,3,6- 11A-6 57.17 (ddd, 7= 8.4, 5.6. 2.4 trifluorophenyl)iminod

[0944] F' T^'- / TBr Hz, 1H), 1.43 (s, 18H) ppm.

[0945] icarbonate

[0946] tert-butyl (tert- butoxycarbonyl)(4,5- XT 479 [M+Na]+; RT: 2.51 min.

[0947] 11A-7F

[0948] difluoro-3- Boc2N'^'Y / T (LC-MS Method 4).

[0949] 1 iodopyridin-2- yl)carbamate

[0950] Br tert-butyl (3-bromo- 430, 432 [M+Na]+; RT: 2.71 11A-8 2,5- £[' difluoropheny 1) (tert- min. (LC-MS Method 4). F ''^ NBOC2

[0951] butoxycarbonyl)carba

[0952]

[0953] 70

[0954] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[0955] mate

[0956] F tert-butyl (5-bromo- 2,3- 408, 410 [M+Na]+; RT: 2.71 11A-9 difluorophenyl) (tert- min. (LC-MS Method 4). Br'x^x^x^x'N(Boc)2butoxycarbonyl)carba

[0957] mate

[0958] tert-butyl (3-bromo-4- (3-cyano-4- Br

[0959] 11A- fluorophenoxy)-5- 561, 563 [M+NaJ+: RT: 2.29 fluoro-2- 10 XJ I I min. (LC-MS Method 5).

[0960] F'^- / ^NBOC2methylphenyl)(tert- butoxycarbonyl)carba

[0961] mate

[0962]

[0963] Intermediate 11B: Tert-butyl (4-bromo-2,3-dichloro-5-((3-cyano-4-fluorophenyl)(hydroxy)methyl)-6-fluorophenyl)(tert-butoxycarbonyl)carbamate Cl

[0964] Ck A. Br ^^. F

[0965] I T J T

[0966] (Boc)2N'^yXY^ / ^CN

[0967]

[0968] F OH

[0969]

[0171] To a stirred and chilled (-78 °C) solution of tert-Butyl (4-bromo-2,3-dichloro-6-fluorophenyl)(tert-butoxycarbonyl)carbamate (2.20 g, 4.79 mmol) in tetrahydrofuran (30 mL) was added lithium diisopropylamide (2.9 mL, 5.75 mmol). The mixture was stirred at -78 °C for 1 hour, then treated with 2-fluoro-5-formyl-benzonitrile (0.75 g, 5.03 mmol). The mixture was stirred at -78 °C for 1 hour, quenched with saturated ammonium chloride (90 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic extracts were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-25% ethyl acetate in petroleum ether) to give the title compound (0.70 g, 24 %) as a solid. MS (ESI): 507, 509 m / z [M+H-Boc]+; purity: 57% (254 nm); retention time: 2.72 minutes (LC-MS Method 4).

[0970]

[0172] The following intermediates were prepared based on the same procedures as described for intermediate 11 B.

[0971] Inter.

[0972] MS m / z [M+H] No. Structure Name+

[0973]

[0974] 71

[0975] IPTS / 200162816.3 Attorney Docket No. SLI-021WO OH F tert-butyl (4-bromo-3-chloro-5-((3- 491, 493 [M+H- cyano-4- 11B-1 X I X I fluorophenyl) (hydroxy)methyl)-2,6- Boc]+; RT: 2.20

[0976] Br-^Y^F difluorophenyl)(tert- S’ o min. (LC-MS 8 a Cl butoxycarbonyl)carbamate

[0977] i H - -n Method 2).

[0978] tert-butyl (3-bromo-4-((3-cyano-4- 539, 541 [M+H]+; RT: fluorophenyl)(hydroxy)methyl)-5- 11B-2 " "1 > 2.41 min. (LC-MS ) 2 S’ o o> / — fluorophenyl)(tert- / / 1i ^_ _ Method 4).

[0979] butoxycarbonyl)carbamate

[0980] Ox )

[0981] Cl tert-butyl (4-bromo-2,3-dichloro-5- CL. Jx. Br zx _ 2 - 2 -F ((3-cyano-4- 507, 509 [M+H-Boc]+; 11B-3 fluorophenyl)(hydroxy)methyl)-6- RT: 2.72 min. (LC-MS I X I I

[0982] (Boc)2N'x^Yxyx^ / ^CN fluorophenyl)(tert- Method 4).

[0983] F OH butoxycarbonyl)carbamate

[0984] OH Br

[0985] tert-butyl (3-bromo-4-((3-cyano-4- NC-^x^ Jx^Jx / F 475, 477 [M+H-Boc]+;

[0986] fluorophenyl)(hydroxy)methyl)-2,5,6- 11B-4 X I I I RT: 2.53 min. (LC-MS F y^NEflCj trifluoropheuy 1) (tertMethod 4). F butoxy carbonyl)carbainate

[0987] z

[0988] F OH u_

[0989] BOC2N XJ'^ / J^ZFX / CN tert-butyl (4-bromo-3-((3-cyano-4- 475, 477 [M+H-Boc]+; fluorophenyl)(hydroxy)methyl)-2,5,6- 11B-5 X I I I RT: 2.09 min. (LC-MS F^' y^ B / r z '-'' T triflu oropheny 1) (tert- O < u-— Method 4). F butoxycarbonyl)carbamate

[0990] OH Br tert-butyl (3-bromo-4-((3-cyano-4- z 457, 459 [M+H]+; RT:

[0991] fluorophenyl)(hydroxy)methyl)-2,5- 11B-6 NC. N < xx A / 'V F

[0992] o 2.53 min. (LC-MS 00 o X I I I difluorophenyl)(tert- F'"^'^ F'^^'NBOC2Method 4).

[0993] butoxycarbonyl)carbamate

[0994] tert-butyl (5-bromo-4-((3-cyano-4- fluorophenyl)(hydroxy)methyl)-2,3- difluorophcnyl)(tcrt- butoxycarbonyl)carbamate 457, 459 fM+H-Boc]+; 11B-7 RT: 2.53 min. (LC-MS and

[0995] Method 4). tert-butyl (5-bromo-4-((3-cyano-4- fluorophenyl)(hydroxy)methyl)-2,3- difluorophenyl)carbamate

[0996]

[0997] Intermediate 11C: (3-(Bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(3-cyano-4-fluorophenyl)methyl acetate

[0998] Cl

[0999] Clx Jx;., Rr ^-Xx, F

[1000] (BOC)2N J T J T

[1001]

[1002] F OAc

[1003]

[0173] To a stirred solution of tert-butyl (4-bromo-2,3-dichloro-5-((3-cyano-4-fluorophenyl)(hydroxy)methyl)-6-fluorophenyl)(tert-butoxycarbonyl)carbamate (Intermediate

[1004] 72

[1005] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1006] 1 IB, 0.95 g, 1.56 mmol) in dichloromethane (20 mL) was added acetyl chloride (0.13 mL, 1.87 mmol) and triethylamine (0.65 mL, 4.69 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (70 mL). The combined organic phase was washed with water (30 mL), brine (30 mL), dried over sodium sulphate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-15% ethyl acetate in petroleum ether) to give the title compound (0.80 g, 78.8%) as a solid. MS (ESI): 549, 551 m / z [M+H-Boc]+; purity: 97% (254 nm); retention time: 2.45 minutes (LC-MS Method 2).

[1007]

[0174] The following intermediates were prepared based on the same procedures as described for intermediate 11C.

[1008] Inter.

[1009] No. Structure Name MS m / z [M+H]+

[1010] OAc F (3-(bis(tert-NCXX^XJXJX, N(BOC)2butoxycarbonyl)amino)-6- 655, 657 [M+Na]+; RT: 11C-1 I J I T bromo-5-chloro-2,4- difluorophenyl)(3-cyano-4- 2.25 min. (LC-MS Cl fluorophenyl)methyl acetate Method 5).

[1011] Br OAc (4-(bis(tert- butoxycarbonyl)amino)-2- 581, 583 [M+H]+; RT: 2.29 11C-2

[1012] bromo-6-fluorophenyl)(3-cyano- min. (LC-MS Method 7). (BOC)2N X ''J / XT CI

[1013] 4-fluorophenyl)methyl acetate

[1014] Cl (3-(bis(tert- butoxycarbonyl)amino)-6- 549, 551 [M+H-Boc]+; RT: 11C-3 I T J I bromo-4,5-dichloro-2- 2.45 min. (LC-MS Method (BOC)2N'X^ / X / TN fluorophenyl)(3-cyano-4- 4).

[1015] F OAc fluorophcnyl)mcthyl acetate

[1016] OAc Br (4-(bis(tert- butoxycarbonyl)amino)-2- I T I T 639, 641 [M+Na]+; RT: 2.66 11C.-4 bromo-3,5,6-trifluorophenyl)(3- min. (LC-MS Method 4). F'"X'^ Fxy^'NBoc2cyano-4-fluorophenyl)methyl

[1017] F acetate

[1018] OAc F (3-(bis(tcrt-N BOC2 butoxycarbonyl)amino)-6- 639, 641 [M+Na]+; RT: 2.67 11C-5 I T I T bromo-2, 4, 5-tri fluorophen yl)(3- F^^ Br 'Y'' T min. (LC-MS Method 4).

[1019] cyano-4-fluorophcnyl)mcthyl

[1020] F acetate

[1021] (4-(bis(tert- OAc Br

[1022] butoxycarbonyl)amino)-2- 621, 623 [M+Na]+; RT: 2.66 11C-6 bromo-3,6-difluorophenyl)(3- min. (LC-MS Method 4).Fn TL, cyano-4-fluorophenyl)methyl

[1023] acetate

[1024]

[1025] 73

[1026] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1027] (4-(bis(tcrt- F OAc butoxycarbonyl)amino)-6- bromo-2,3-difluorophenyl)(3- cyano-4-fluorophcnyl)mcthyl

[1028] (BocJjN'^^'Br '"'^'"'F acetate

[1029] 621, 623 [M+Na]+; RT: 2.49 11C-7

[1030] F OAc And min. (LC-MS Method 7).

[1031] (6-bromo-4-((tert- butoxycarbonyl)amino)-2,3- BocHN^'^ Br ^^F

[1032] difluorophenyl)(3-cyano-4- fluorophenyl)methyl acetate

[1033]

[1034] Intermediate 11D: (3-(Bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(3-carbamothioyl-4-fluorophenyl)methyl acetate

[1035] ci

[1036]

[1037] F OAc S

[1038]

[0175] To a stirred solution of sodium hydrosulfide (0.30 g, 3.69 mmol) and magnesium chloride hexahydrate (0.38 g, 1.85 mmol) in N, N-dimethylformamide (15 mL) was added (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(3-cyano-4-fluorophenyl)methyl acetate (Intermediate 11C, 0.80 g, 1.23 mmol). The mixture was stirred at room temperature for 1 hour, diluted with water (45 mL), and extracted with ethyl acetate (45 mL x 2). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (40 g column, eluting with ethyl acetate in petroleum ether 0-60%) to give the title compound (0.80 g, 95 %) as a yellow solid. MS (ESI): 705, 707 in / z [M+Na]+; purity: 96% (214 nm); retention time: 2.68 minutes (LC-MS Method 4).

[1039]

[0176] The following intermediates were prepared based on the same procedures as described for intermediate HD.

[1040] Inter.

[1041] No. Structure Name MS m / z [M+H]+Br OAc S

[1042] (3-(bis(tert-butoxycarbonyl)ainino)- 567, 569 [M+H-Boc]+; 6-bromo-5-chloro-2,4- 11D-1

[1043] difluorophenyl)(3-carbamothioyl-4- RT: 2.13 min. (LC-MS fluorophenyl)methyl acetate

[1044] N(BOC)2Method 5).

[1045] S

[1046] 586, 588 [M+Na]+; RT: Boc tert-butyl (4-bromo-5-((3- 11D-2

[1047] R2.23 min. (LC-MS Y T nnH2carbamothioyl-4- Boc

[1048] fluorophenyl)thio Method 2).)thiazol-2-yl)(tert-

[1049]

[1050] 74

[1051] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1052] butoxycarbonyl)carbamatc

[1053] Br OAc S (4-(bis(tert-butoxycarbonyl)amino)- I 2-bromo-6-fluorophenyl)(3- 637, 639 [M+Na]+; RT: 11D-3 N> 2.23 min. (LC-MS xV\x^NH2carbamothioyl-4- Method 7). fluorophenyl)methyl acetate

[1054] t Cl (3-(bis(tert-butoxycarbonyl)amino)- CI^ / L. Br 705, 707 [M+Na]+; RT:

[1055] 6-bromo-4,5-dichloro-2- 11D-4 2.68 min. (LC-MS fluorophenyl)(3-carbamothioyl-4- Method 4). F OAc S fluorophenyl)methyl acetate

[1056] z -n

[1057] I F NH25-(4-amino-6-bromo-2,3- 377, 379 [M+H]+; RT: 11D-5 difluorophenoxy)-2- 1.86 min. (LC-MSFXX°TX^Sfluorobenzothioamide Method 2).

[1058] F S tert-butyl (5-bromo-4-(3- 477, 479 [M+II-Boc]+; carbamothioyl-4-fluorophenoxy)-2,3- 11D-6FYy Vy ^2RT: 1.86 min. (LC-MS difluorophenyl)(tert- (Boc)2N'Xx^XBr Method 2).

[1059] biitoxycarbonyl)carbamate

[1060] F OAc S (4-(bis(tert-butoxycarbonyl)amino)- 2-bromo-3,5,6-trifluorophenyl)(3- 673, 675 [M+Na]+; RT: 11D-7 2.45 min. (LC-MS Boc2N' y / <Br carbamothioyl-4- Method 4). F fluorophenyl)methyl acetate

[1061] F OH S tert-butyl (4-bromo-3-((3- carbamothioyl-4- 631, 633 [M+Na]+; RT: 11D-8 r if ii iNfluorophenyl)(hydroxy)methyl)- 2.18 min. (LC-MS F'^ y^Br ^^^FH2

[1062] 2,5,6-trifluorophenyl)(tert- Method 3). F

[1063] butoxycarbonyl)carbamate

[1064] Br S

[1065] 5-(3-amino-6-bromo-2,4,5- 393, 395 [M+H]+; RT: 11D-9FXXXXTNH2trifluorobenzyl)-2- 2.18 min. (LC-MS F^y ’Xp fluorobenzothioamide Method 7).

[1066] NH2

[1067] Br S

[1068] 5-(3-amino-2-bromo-4,5,6- 395, 397 [M+H]+; RT: LID-10 trifluorophenoxy)-2- 2.17 min. (LC-MS F" y"' T fluorobenzothioamide Method 7).

[1069] F S OAc Br (4-(bis(tert-butoxycarbonyl)aimno)- 2-bromo-3,6-difluorophenyl)(3- 655, 657 [M+Na]+; RT: 11D-11 2.45 min. (LC-MSF'-W<NBOC2carbamothioyl-4- Method 4). fluorophenyl)methyl acetate

[1070] 5-((4-amino-6-bromo-2,3- 393, 395 [M+H]+; RT: 11D-12 difluorophenyl)thio)-2- 1.77 min. (LC-MS fluorobenzothioamide Method 11).

[1071]

[1072] 75

[1073] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1074] (4-(bis(tcrt-butoxycarbonyl)amino)- NH2OAC F 6-bromo-2,3-difluorophenyl)(3- carbamothioyl-4- 655, 657 [M+Na]+; RT: fluorophenyl)methyl acetate

[1075] F '^ Br "^ N(BOC)22.49 min. (LC-MS 11D-13 And Method 7) and NH2OAC F

[1076] (6-bromo-4-((tert- 555, 557 [M+Na]+; (LC- butoxycarbonyl)amino)-2,3- MS Method 7). difluorophenyl)(3-carbamothioyl-4- tluorophenyl)methyl acetate

[1077] S Br 5-(4-amino-2-bromo-3-chloro-6- A / L,ci 393, 395 [M+II]+; RT: 11D-14 H2N Ap |T Y p fluorophenoxy)-2- 2.15 min. (LC-MS F^^5^ F^^'" NH2fluorobenzothioamide Method 7).

[1078] S Br 5-(4-amino-2-bromo-3,6- 377, 378 [M+H]+; RT: 11D-15H2NFdifluorophenoxy)-2- 1.87 min. (LC-MS F'>^><'NH2fluorobenzothioamide Method 3).

[1079] S Br 5-((2-bromo-6-fluoro-4-((4- A.. S,. A / methoxybenzyl)oxy)-3- 510, 512 [M+H]+; RT: 11D-16 1.87 min. (LC-MS methylphenyl)thio)-2- F^^^ F'X''^''OPMB Method 3).

[1080] fluorobenzothioamide

[1081] S Br 5-(2-bromo-3,6-difluoro-4- 392, 394 [M+H]+; RT: 11D-17 methoxyphenoxy)-2- 1.77 min. (LC-MS F'^A fluorobenzothioamide Method 2).

[1082]

[1083] Intermediate 12: Tert-butyl (4-bromo-5-((3-cyano-4-fluorophenyl)thio)thiazol-2-yl)(tert-butoxycarbonyl)carbamate

[1084]

[1085]

[0177] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(5-((3-cyano-4-fluorophenyl)thio)thiazol-2-yl)carbamate (Intermediate 12B, 8.20 g, 18.2 mmol) in N, N-dimethylformamide (100 mL) was added N-bromosuccinimide (6465 mg, 36.3 mmol). The reaction was stirred at room temperature for 24 hours, then treated with more N-bromosuccinimide (6465 mg, 36.3 mmol), and stirred for another 24 hours. The mixture was quenched with 200 mL of water and extracted with ethyl acetate (200 mL x 2). The combined organic phases were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (eluting with 0-40% of ethyl acetate in petroleum ether) to give the title compound (7.00 g, 13.2 mmol, 73 %). MS (ESI): 554, 552 m / z [M+Na]+; purity: 90% (254 nm); retention time: 2.37 minutes (LC-MS Method 2).

[1086] 76

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[1088] Intermediate 12A: 5-((2-Aminothiazol-5-yl)thio)-2-fluorobenzonitrile

[1089]

[1090] CN

[1091]

[0178] To a stirred solution of 5-bromothiazol-2-amine hydrobromide (19.01 g, 73.1 mmol) in N, N-dimethylformamide (50mL) were added 2-fluoro-5-mercaptobenzonitrile (11.20 g. 73.1 mmol) and cesium carbonate (71473 mg, 219 mmol). The mixture was stirred at 50 °C for 10 minutes, poured into water (1000 mL), and extracted with ethyl acetate (200 mL x 2). The combined organic layers were washed with water (200 mL), brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (eluting with 0~70 % ethyl acetate in petroleum ether) to give the title compound (5.30 g, 21.1 mmol, 29 % yield). MS (ESI): 252 m / z [M+H]+; purity: 94% (254 nm); retention time: 1.68 minutes (LC-MS Method 2).

[1092] Intermediate 12B: Tert-butyl (tert-butoxycarbonyl)(5-((3-cyano-4-fluorophenyl)thio)thiazol-2-yl)carbamate

[1093]

[1094]

[0179] To a stirred solution of 5-((2-aminothiazol-5-yl)thio)-2-fluorobenzonitrile (Intermediate 12A, 5.30 g, 21.1 mmol) in toluene (100 mL) were added 4-dimethylaminopyridine (515 mg, 4.22 mmol) and di -tert-butyl dicarbonate (11 mL, 46.4 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 2 hours. The mixture was concentrated. The residue was purified by flash chromatography (120 g silica gel column, eluting with 0-40% ethyl acetate in petroleum ether) to give the title compound (8.20 g, 18.2 mmol, 86 %) as an oil. MS (ESI): 474 m / z [M+Na]+; purity: 96% (214 nm); retention time: 2.28 minutes (LC-MS Method 2).

[1095] Intermediate 13: Tert-butyl 2-(3,5-difluorophenyl)-7-hydroxy-2,6,6-trimethylheptanoate

[1096]

[1097]

[0180] To a stirred solution of tert-butyl 7-[tert-butyl(dimethyl)silyl]oxy-2-(3,5-difluorophenyl)-2,6,6-trimethyl-heptanoate (Intermediate 13 A, 50.00 g, 106

[1098] 77

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[1100] mmol) in tetrahydrofuran (500 mL) was added tetra-n-butylammonium fluoride (33328 mg, 127 mmol) at room temperature. The mixture was stirred for 2 hours, quenched with water (300 mL), and extracted with ethyl acetate (300 mL x 3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (120 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (30.2 g,84.8 mmol, 80) as a liquid. MS (ESI): 379 m / z fM+Na]+; purity: 93% (214 nm); retention time: 2.22 minutes (LC-MS Method 2).

[1101] Intermediate 13A: Tert-butyl 7-[tert-butyl(dimethyl)silyl]oxy-2-(3,5-difluorophenyl)-2,6,6-trimethyl-heptanoate

[1102]

[1103]

[0181] To a stirred and chilled (-78 °C) solution of tert-butyl 2-(3,5-difluorophenyl)propanoate (50.00 g, 206 mmol) in tetrahydrofuran (500 mL) was slowly added lithium diisopropylamide (2.0 M in tetrahydrofuran, 135 mL, 270 mmol) under nitrogen. The mixture was stirred for 1 hour, treated with tert-butyl-(5-iodo-2,2-dimethyl-pentoxy)-dimethyl-silane (1.10 eq, 90.00 g, 253 mmol) drop wise, and stirred at this temperature for another 5 hours. The reaction mixture was quenched with water (300 mL), and extracted with ethyl acetate (300 mL x 3), The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (120 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (186.00 g, 181 mmol, 88%) as a liquid. MS (ESI): 471 nz / z [M+H]+; purity: 46% (214 nm); retention time: 2.06 minutes (LC-MS Method 2).

[1104] Intermediate 14: 2-(3,5-Difluorophenyl)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethylheptanoic acid

[1105]

[1106]

[0182] To a stirred solution of tert-butyl 2-(3,5-difluorophenyl)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethylheptanoate (Intermediate 9D-1, 10.00 g, 22.3 mmol) in 1,4-dioxane (100 mL)

[1107] 78

[1108] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1109] was added hydrochloric acid (0.68 mL, 22.3 mmol) at room temperature. The mixture was stirred for 2 hours, quenched with water (300 mL), and extracted with ethyl acetate (lOOmL x 3). The com Tbi toned organic extracts were washed with brine, dried over sodium sulfate, filtered,

[1110] Z ^TT^^

[1111] and concentrated. The residue was purified by flash chromatography (120 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (9.00 g, 14.2

[1112] O CD

[1113] mmol, 64 %) as liquid. MS (ESI): 415 m / z [M+Na]+; purity: 62% (214 nm); retention time: 1.78 minutes (LC-MS Method 2).

[1114] A v

[1115] Intermediate 15: 5-(4-Amino-2-bromo-6-fluorophenoxy)-2-fluorobenzonitrile

[1116]

[0183] To a stirring solution of 5-(2-Bromo-6-fluoro-4-nitrophenoxy)-2-fluorobenzonitrile (3.10 g, 8.73 mmol) in acetic acid (45 mL) was added iron (2.44 g, 43.7 mmol). The mixture was stirred at room temperature for 2 hours, filtered through a pad of Celite, and the filter cake rinsed with ethyl acetate (90 mL). The combined filtrates were concentrated. The residue was dissolved in ethyl acetate (150 mL), washed with saturated aqueous sodium bicarbonate (40 mL), water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography to give the title compound (2.80 g, 8.61 mmol, 98 %) as a solid. MS (ESI): 325, 327 m / z [M+H]+; purity: 62% (214 nm); retention time: 1.95 minutes (LC-MS Method 2).

[1117]

[0184] The following intermediate was prepared based on the procedures described for Intermediate 15.

[1118] Inter.

[1119] Structure Name MS

[1120] No. m / z [M+HJ+

[1121] 5-(4-amino-6-bromo-2,3- 343, 345 [M+H]+; 15-1 difluorophenoxy)-2- RT: 1.37 min. (LC- fluorobeiizonitrile MS Method 1). Br

[1122] 359, 361 [M+H]+; 5-(4-amino-2-bromo-3-chloro-6- 15-2 TJ I T RT: 1.99 min. (LC- fluorophenoxy)-2-fluorobenzonitrile

[1123] MS Method 5). F^'"^ F'^ / ^NH2

[1124] Br

[1125] 339, 341 [M+H]+; 5-(4-amino-2-bromo-6-fluoro-3- 15-3 TJ J T RT: 1.99 min. (LC- methylphenoxy)-2-fluorobenzonitrile

[1126] MS Method 2).

[1127]

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[1130] 5-(4-amino-2-bromo-3,6- 343, 345 [M+H]+; difluorophenoxy)-2- RT: 1.99 min. (LC- fluorobenzonitrile MS Method 3).

[1131]

[1132] Intermediate 15A: 5-(2-Bromo-6-fluoro-4-nitrophenoxy)-2-fluorobenzonitrile

[1133]

[1134]

[0185] To a stirred solution of 2-fluoro-5-hydroxybenzonitrile (1.43 g, 10.5 mmol) and 1-bromo-2,3-difluoro-5-nitrobenzene (2.37 g, 9.96 mmol) in N, N-dimethylformamide (45 mL) was added potassium carbonate (2.75 g, 19.9 mmol). The mixture was stirred at room temperature for 16 hours, quenched with water (150 mL), and extracted with ethyl acetate (60 mL x 3). The combined organic extracts were washed with water (40 mL), brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (3.10 g, 8.73 mmol, 87 %) as a light yellow solid. MS (ESI): 355, 357 m z [M+H]+; purity: 96% (214 nm); retention time: 2.04 minutes (LC-MS Method 2).

[1135]

[0186] The following intermediates were prepared based on the procedures described for Intermediate 15.

[1136] Inter.

[1137] Structure Name MS m / z [M+H] No.+

[1138] 373, 375 [M+H]+; 5-(6-bromo-2,3-difluoro-4- 15 A- 1 RT: 1.41 mill. (LC- nitrophenoxy)-2-fluorobenzonitrile

[1139] O2N' I^ / TxBr I T MS Method 7).

[1140] Br

[1141] NC_^O_X / CI 5-(2-bromo-3-chloro-6-fluoro-4- No MS signal 15A-2

[1142] nitrophcnoxy)-2-fluorobcnzonitrilc observed F Tj ^ F I '^^TNO2

[1143] Br

[1144] 369, 371 [M+H]+; 5-(2-bromo-6-fluoro-3-methyl-4- 15A-3 T y y T RT: 2.20 min. (LC- nitropheuoxy)-2-fluorobenzouitrile

[1145] MS Method 5). F F "'~''' LO2

[1146] Br

[1147] NC^^Q^ A / F 373, 375 [M+H]+;

[1148] 5-(2-bromo-3,6-difluoro-4- 15A-4 T y I T RT: 2.09 min. (LC- nitrophenoxy)-2-fluorobenzonitrile

[1149] MS Method 3). F'^"-"^ F'^^''NO

[1150]

[1151] 2

[1152] Intermediate 16: Tert-Butyl((6-iodo-2,2-dimethylhexyl)oxy)dimethylsilane

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[1155] OTBS

[1156]

[1157]

[0187] To a stirred solution of tert-butyl((6-iodo-2,2-dimethylhexyl)oxy)dimethylsilane (Intermediate 16C, 27.00 g, 96.8 mmol) in acetone (300 mL) was added sodium iodide (43.50 g, 290 mmol) at room temperature. The mixture was stirred at 60 °C for 16 hours. The solvent was evaporated. The residue was dissolved in ethyl acetate (500 mL), washed with water (2 x 300 mL), dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by flash chromatography (330 g silica gel column, petroleum ether) to give the title compound as acolorless oil (32.00 g, 89.3%).1H NMR (400 MHz, CDCl3) 3.25-3.13 (m, 4H), 1.80-1.68 (m, 2H), 1.39-1.27 (m, 2H), 1.23-1.16 (m. 2H), 0.87 (s, 9H), 0.80 (s, 6H), -0.01 (d, J = 7.0 Hz, 6H).

[1158]

[0188] The following intermediate was prepared based on the procedures described for Intermediate 16.

[1159] Inter.

[1160] Structure Name MS m / z [M+H] No.+tert-butyl((5-iodo-2,2- 16-1 357

[1161] dimethylpentyl)oxy)diinethylsilane

[1162]

[1163] Intermediate 16A: Methyl 6-chloro-2,2-dimethyl-hexanoate

[1164] O

[1165]

[1166]

[0189] To a stirred and cooled (-78 °C) solution of methyl 2-methylpropanoate (10.30 g, 101 mmol) in tetrahydrofuran (200 mL) was added lithium diisopropylamide (50.4 mL, 101 mmol) drop wise. After stirring at -78 °C for 1 hour, l-chloro-4-iodo-butane (20.00 g, 91.5 mmol) was added. The mixture was stirred at -78 °C for 1 hour and then allowed to warm to room temperature and stirred for additional 16 hours. The reaction was quenched with IM hydrochloric acid (200 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic extracts were washed with water, brine, dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by flash silica gel chromatography (eluting with 50% ethyl acetate in petroleum ether) to give the title compound as a light-yellow oil (16.00 g, 82.9%). MS (ESI): 193 m z [M+H]+.

[1167] The following intermediate was prepared based on the procedures described for Intermediate 16 A.

[1168] 81

[1169] IPTS / 200162816.3 Attorney Docket No. SLI-021WO Inter.

[1170] Structure Name MS m / z [M+H] No.+o

[1171] 16A-1 methyl 5-chloro-2,2-dimethylpentanoate 179

[1172]

[1173] Intermediate 16B: 6-Chloro-2,2-dimethylhexan-l-ol

[1174]

[1175]

[0190] To a stirred and cooled (-78 °C) solution of methyl 6-chloro-2,2-dimethyl-hexanoate (Intermediate 16A, 24.00 g, 125 mmol) in tetrahydrofuran (200 mL) was added lithium aluminum hydride (7.09 g, 187 mmol) dropwise over 30 minutes. The mixture was stirred for 3 hours at -78 °C, then quenched with 7 mL of water, warmed to 0 °C, followed by 7 mL of 15% aqueous sodium hydroxide, and finally 21 mL of water. The mixture was stirred at room temperature for 15 minutes and then filtered. The filtrate was concentrated to give the title compound as an oil (18.00 g, 87.8%). ‘H NMR (400 MHz, CDCh) d 3.55 (t, J = 6.7 Hz, 2H), 3.53 (s, 2H), 1.80 - 1.73 (m, 2H), 1.4-1.39 (m, 2H), 1.28-1.26 (m, 2H), 0.86 (s, 6H) ppm.

[1176]

[0191] The following intermediate was prepared based on the procedures described for Intermediate 16 A.

[1177] Inter.

[1178] Structure Name MS m / z [M+H] No.+O

[1179] 16A-1 methyl 5-chloro-2,2-dimethylpentanoate 179

[1180]

[1181] Intermediate 16C: Tert-Butyl-(6-chloro-2,2-dimethyl-hexoxy)-dimethyl-silane

[1182] Cl

[1183]

[1184]

[0192] To a stirred solution of 6-chloro-2,2-dimethyl-hexan-l-ol (Intermediate 16B, 18.00 g, 109 mmol) in dichloromethane (300 mL) was added tert-butyldimethylsilyl chloride (19.80 g, 131 mmol) and imidazole (14.9 g, 219 mmol). The mixture was stirred at room temperature for 2 hours, then diluted with dichloromethane (200 mL). The solution was washed with water (2 x 100 mL), brine, dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by flash chromatography (120 g silica gel column, petroleum ether) to give the title compound as an oil (27.00 g, 89%).1H NMR (400 MHz, CDCh) d 3.52 (t, J = 6.7 Hz,

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[1187] 2H), 3.21 (s, 2H), 1.76-1.68 (m, 2H), 1.35 (dt, 7 = 10.7, 7.9 Hz, 2H), 1.23-1.18 (m, 2H), 0.87 (s, 9H), 0.80 (d, J= 5.8 Hz, 6H), -0.01 (d, J= 6.6 Hz, 6H) ppm.

[1188] Intermediate 17: 4-Bromo-2,3,5-trifluoro-6-nitropyridine

[1189]

[1190] Br

[1191]

[0193] A mixture of hydrogen peroxide (10 mL, 97.9 mmol) in sulfuric acid (10 mL, 188 mmol) was stirred at 0 °C for 10 minutes, then treated with 4-bromo-3,5,6-trifluoro-pyridin-2-amine (902 mg, 3.97 mmol) at 0 °C. The mixture was stirred at room temperature overnight, poured into ice water, basified with saturated aqueous sodium bicarbonate to pH ~ 8, and extracted with ethyl acetate (40 mL x 3). The combined organic extracts were washed with brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (12 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (388 mg, 1.51 mmol, 38 %) as a yellow oil. MS (ESI): Not observed m / z [M+H]+; purity: 98% (214 nm); retention time: 2.10 minutes (LC-MS Method 4).

[1192] Intermediate 17A: 4-Bromo-3,5,6-trifluoropyridin-2-amine

[1193]

[1194] Br

[1195]

[0194] A mixture of 4-bromo-2,3,5,6-tetrafluoropyridine (908 mg, 3.95 mmol) in ammonia (3.9 mL, 15.8 mmol) was stirred at 100 °C for 1 hour in a microwave reactor. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (40 mL x 2). The combined organic extracts were washed with brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (860 mg, 3.79 mmol, 96 %) as a yellow solid. MS (ESI): 227, 229 m / z. [M+H]+; purity: 91% (214 nm); retention time: 2.03 minutes (LC-MS Method 4).

[1196] Intermediate 18: 5-(3-Amino-6-bromo-2,4,5-trifluorobenzyl)-2-fluorobenzonitrile

[1197]

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[1200]

[0195] To a stirred solution of (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-2,4,5-trifluorophenyl)(3-cyano-4-fluorophenyl)niethyl acetate (Intermediate 11C-5, 1.5 g, 2.43 mmol) in dichloromethane (15 mL) was added triethylsilane (1.6 mL, 9.72 mmol) and boron trifluoride etherate (1.2 mL, 9.72 mmol) at 0 °C under N2 atmosphere. The mixture was stirred at room temperature for 16 hours, diluted with water (50 mL), and extracted with ethyl acetate (50 mL x 2). The combined organic extracts were washed with water, and brine, dried over anhydrous sodium sulphate, filtered, and concentrated. The residue was purified by flash chromatography (80 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (650 mg, 74%) as a yellow oil. MS (ESI): 359, 361 m / z [M+H]+; purity: 71% (254 nm); retention time: 1.96 minutes (LC-MS Method 5).

[1201] Intermediate 19: 4,5-Difluoro-3-iodopyridin-2-amine

[1202] F\1

[1203] y— NH2

[1204]

[1205] \=N

[1206]

[0196] A mixture of 4,5-difhioropyridin-2-amine (1.00 eq, 500 mg, 3.84 mmol) and N-iodosuccinimide (1729 mg, 7.69 mmol) in trifluoroacetic acid (10 mL, 130 mmol) was stirred at room temperature for 2 hours, then overnight at 40 °C, and basified with saturated sodium bicarbonate aqueous solution to pH ~ 8, and extracted with ethyl acetate (60 mL x 3). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (12 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to afford the title compound (337 mg, 1.32 mmol, 34 %) as a yellow oil. MS (ESI): 257 m / z [M+H]+; purity: 97% (254 nm); retention time: 1.90 minutes (LC-MS Method 4).

[1207] Intermediate 20: 5-(3-Amino-2-bromo-4,5,6-trifluorophenoxy)-2-fluorobenzonitrile

[1208] Br

[1209]

[1210]

[0197] To a stirred solution of 5-(5-amino-2,3,4-trifluoro-phenoxy)-2-fluoro-benzonitrile (Intermediate 20H, 230 mg, 0.815 mmol) in acetic acid (5 mL) was added N-bromosuccinimide (174 mg, 0.978 mmol). This mixture was stirred at room temperature overnight, diluted with water (30 mL), and extracted with ethyl acetate (10 mL x 3). The combined organic extracts

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[1213] were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (4.0 g silica gel column, eluting with 0-40% ethyl acetate in petroleum ether) to afford the title compound (280 mg, 0.775 mmol, 95 %) as a light yellow solid. MS (ESI): 361, 363 m / z [M+H]+; purity: 94% (214 nm); retention time: 2.35 minutes (LC-MS Method 4).

[1214] Intermediate 20A: l,2,3-Trifluoro-4-((4-methoxybenzyl)oxy)benzene

[1215] „PMB

[1216]

[1217]

[0198] To a stirred solution of 2,3,4-trifluorophenol (5.00 g, 33.8 mmol) in N, N-dimethylformamide (40 mL) was added 4-methoxybenzyl chloride (5.5 mL, 40.5 mmol), followed by potassium carbonate (7.00 g, 50.6 mmol). The mixture was stirred at room temperature overnight, diluted with water (200 mL), and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (80 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (8.50 g, 31.7 mmol, 94 %) as a white solid.XH NMR (500 MHz, DMSO-de) S 7.40-7.37 (m, 2H), 7.29-7.23 (m, 1H), 7.15-7.10 (m, 1H), 6.97-6.94 (m, 2H), 5.11 (s, 2H), 3.76 (s, 3H) ppm.

[1218] Intermediate 20B: (2,3,4-Trifluoro-5-((4-methoxybenzyl)oxy)phenyl)boronic acid

[1219] F

[1220]

[1221] OH

[1222]

[0199] To a stirred solution of l,2,3-Trifluoro-4-((4-methoxybenzyl)oxy)benzene (Intermediate 20A, 6.50 g, 24.2 mmol) in tetrahydrofuran (50 mL) was added lithium diisopropylamide (2.0 M solution in tetrahydrofuran / n-heptane) (13.3 mL, 26.7 mmol) at -78 °C. The mixture was stirred at -78 °C for 0.5 hours, treated with trimethyl borate (3.2 mL, 29.1 mmol), and stirred at -78 °C for 1.5 hours. The mixture was quenched with saturated ammonium chloride (300 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (80 g silica gel column,

[1223] 85

[1224] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1225] eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (4.00 g. 12.8 mmol, 53 %) as a white solid. ‘H NMR (500 MHz, DMSO-d6) d 8.46 (s, 2H), 7.39-7.37 (m, 2H), 7.24-7.21 (m, 1H), 6.97-6.95 (m, 2H), 5.10 (s, 2H), 3.76 (s, 3H) ppm.

[1226] Intermediate 20C: 2,3,4-Trifluoro-5-((4-methoxybenzyl)oxy)phenol

[1227] F

[1228]

[1229]

[0200] To a stirred solution of (2,3,4-trifluoro-5-((4-methoxybenzyl)oxy)phenyl)boronic acid (Intermediate 20B, 5.00 g, 16.0 mmol) in ethanol (100 mL) was added hydrogen peroxide (8.2 mL, 80.1 mmol). The mixture was stirred at room temperature overnight, diluted with water (300 mL), and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-25% ethyl acetate in petroleum ether) to give the title compound (4.00 g, 14.1 mmol, 88 %) as a white solid. MS (ESI): 307 m / z [M+Na]+; purity: 90% (214 nm); retention time: 2.25 minutes (LC-MS Method 4).

[1230] Intermediate 20D: 2-Fluoro-5-(2,3,4-trifluoro-5-((4-methoxybenzyl)oxy)phenoxy)benzonitrile

[1231]

[1232]

[0201] To a stirred solution of 2,3,4-trifluoro-5-((4-methoxybenzyl)oxy)phenol (Intermediate 20C, 4.80 g, 16.9 mmol) in acetone (150 mL) was added (3-cyano-4-fluorophenyl)(2,4,6-trimethoxyphenyl)-13-iodaneyl 4-methylbenzenesulfonate (10.87 g, 18.6 mmol), followed by potassium carbonate (7.00 g, 50.7 mmol). The mixture was stirred at 50 °C overnight, diluted with water (300 mL), and extracted was ethyl acetate (3 x 150 mL). The combined organic phases were washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (80 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (4.00 g, 9.92 mmol, 59 %) as a white solid. MS (ESI): 404 m / z [M+H]+; purity: 53% (214 nm); retention time: 2.60 minutes (LC-MS Method 4).

[1233] 86

[1234] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1235] Intermediate 20E: 2-Fluoro-5-(2,3,4-trifluoro-5-hydroxyphenoxy)benzonitrile

[1236] F

[1237]

[1238]

[0202] To a stirred solution of 2-fluoro-5-(2,3,4-trifluoro-5-((4-methoxybenzyl)oxy)phenoxy)benzonitrile (Intermediate 20D, 4.00 g, 9.92 mmol) in dichloromethane (15 mL) was added trifluoroacetic acid (5.0 mL, 64.9 mmol). The mixture was stirred at room temperature for 2 hours, the pH was adjusted to ~ 8 with saturated aqueous sodium bicarbonate, and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (40 g silica gel column, eluting with 0-25% ethyl acetate in petroleum ether) to afford the title compound (2.50 g, 8.83 mmol, 89 %) as a white solid. MS (ESI): 284 m / z [M+H]+; purity: 61% (214 nm); retention time: 2.20 minutes (LC-MS Method 4).

[1239] Intermediate 20F: 5-(3-Cyano-4-fluorophenoxy)-2,3,4-trifluorophenyl trifluoromethanesulfonate

[1240] F

[1241]

[1242]

[0203] To a stirred solution of 2-fluoro-5-(2,3,4-trifluoro-5-hydroxy-phenoxy)benzonitrile (Intermediate 20E, 1.70 g, 6.00 mmol) in dichloromethane (20 mL) was added pyridine (0.97 mL, 12.0 mmol), followed by trifluoromethanesulfonic anhydride (1.5 mL, 9.00 mmol) at 0 °C. The mixture was stirred at 0 °C for 2 hours and concentrated. The residue was purified by flash chromatography (80 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (1.50 g, 3.61 mmol, 60 %) as a light-yellow oil. MS (ESI): 416 m / z [M+H]+; purity: 61% (214 nm); retention time: 2.56 minutes (LC-MS Method 4).

[1243] Intermediate 20G: 5-(5-((Diphenylmethylene)amino)-2,3,4-trifluorophenoxy)-2-fluorobenzonitrile

[1244]

[1245] 87

[1246] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1247]

[0204] To a stirred solution of 5-(3-cyano-4-fluorophenoxy)-2,3,4-trifluorophenyl trifluoromethanesulfonate (Intermediate 20F, 1.90 g, 4.58 mmol) in tetrahydrofuran (50 mL) were added benzophenone imine (1.0 mL, 5.95 mmol), cesium carbonate (2.53 g, 7.78 mmol) tris(dibenzylideneacetone)dipalladium(0) (419 mg, 0.458 mmol) and (9,9-dimethyl-9h-xanthene-4,5-diyl)bis(diphenylphosphine) (530 mg, 0.915 mmol). The mixture was stirred at 70 °C under an argon atmosphere for 3 hours, diluted with water (100 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-15% ethyl acetate in petroleum ether) to give the title compound (1.00 g, 2.24 mmol, 49 % yield) as light-yellow oil. MS (ESI): 447 m / z [M+H]+; purity: 55% (214 nm); retention time: 2.86 minutes (LC-MS Method 4).

[1248] Intermediate 20H: 5-(5-Amino-2,3,4-trifluorophenoxy)-2-fluorobenzonitrile

[1249] F

[1250]

[1251] NH

[1252]

[0205] A mixture of 5-(5-((diphenylmethylene)amino)-2,3,4-trifluorophenoxy)-2-fluorobenzonitrile (Intermediate 20G, 1.00 g, 2.24 mmol) in hydrochloric acid / 1,4-dioxane (4.0 M, 10 mL, 40.0 mmol) was stirred at room temperature for 1 hour. The mixture was diluted with water (20 mL), the pH adjusted to ~ 8 with saturated aqueous sodium bicarbonate and extracted with ethyl acetate (3 x 40 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (12 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to afford the title compound (250 mg, 0.886 mmol, 39 %) as a light-yellow solid. MS (ESI): 283 m / z [M+H]+; purity: 88% (214 nm); retention time: 2.26 minutes (LC-MS Method 4).

[1253] Intermediate 21: 5-((4-Amino-6-bromo-2,3-difluorophenyl)thio)-2-fluorobenzonitrile F

[1254] , S

[1255]

[1256]

[0206] To a stirred solution of a mixture of 5-(6-bromo-2,3-difluoro-4-nitro-phenyl)sulfanyl-2-fluoro-benzonitrile and 5-(4-bromo-2,3-difluoro-6-nitro-phenyl)sulfanyl-2-fluoro-benzonitrile (Intermediate 21A, 15 g, 38.6 mmol) in acetic acid (150 mL) was added iron (6.5

[1257] 88

[1258] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1259] g, 77.2mmol). The reaction mixture was stirred at 40 °C under argon for 2 hours, filtered, and concentrated. The residue was dissolved in ethyl acetate (1000 ml), washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was slurried with 10% ethyl acetate in hexane and filtered. The solid was dried in vacuo to afford the title compound (11 g, 30.6 mmol, 80 %). MS (ESI): 359, 361 m / z [M+H]+; purity: 95% (254 nm); retention time: 1.97 minutes (LC-MS Method 2)

[1260] Intermediate 21A: Mixture of 5-((4-bromo-2,3-difluoro-6-nitrophenyl)thio)-2-fluorobenzonitrile and 5-((6-bromo-2,3-difluoro-4-nitrophenyl)thio)-2-fluorobenzonitrile

[1261]

[1262]

[0207] To a stirred suspension of l-bromo-2,3,4-trifluoro-5-nitro-benzene (30.5 g, 119 mmol) and sodium bicarbonate (20 g, 238 mmol) in N, N-dimethylformamide (300 mL) was added dropwise a solution of 2-fluoro-5-mercaptobenzonitrile (16.8 g, 110 mmol) in N, N-dimethylformamide (200 mL) at -60 °C. The reaction mixture was stirred at -60 °C under argon for 1 hour, quenched with water (1000 mL), and extracted with ethyl acetate (500 mL X 3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (330g silica gel column, eluting with 0-40% ethyl acetate in petroleum ether) to give the title product as an inseparable mixture (36.4 g, 93.6 mmol, 93 %, a yellow solid). MS (ESI): 389, 391 m / z [M+H]+; purity: >99% (214 nm); retention time: 3.06 minutes (LC-MS Method 10).

[1263] Intermediate 22: Tert-butyl (3-bromo-4-(3-carbamimidoyl-4-fluorophenoxy)-5-fluoro-2-methylphenyl)(tert-butoxycarbonyl)carbamate

[1264] Br

[1265]

[1266]

[0208] To a stirred solution of tert-butyl (3-bromo-4-(3-cyano-4-fhtorophenoxy)-5-fluoro-2-methylphenyl)(tert-butoxycarbonyl)carbamate (Intermediate 11A-10, 200 mg, 0.371 mmol) in tetrahydrofuran (6 mL) was added lithium bis(trimethylsilyl)amide (IM in tetrahydrofuran, 1.48 mL, 1.48 mmol) at 0 °C. The reaction was stirred at room temperature for 16 hours, quenched with saturated ammonium chloride (20 mL), and extracted with ethyl acetate (50 mL 89

[1267] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1268] x 2). The combined organic extracts were washed with brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give the title compound (200 mg, 0.359 mmol, 97 % yield) as a brown solid. MS (ESI): 556, 558 m / z [M+H]+; purity: 89% (214 nm); retention time: 1.62 minutes (LC-MS Method 5).

[1269] Intermediate 23: Methyl (S)-3-(3-((R)-l-bromo-8-((2-((tert-butyldiphenylsilyl)oxy)ethyl)sulfonyl)-3,7,7-trimethyl-2-oxooctan-3-yl)phenyl)-2-methylpropanoate

[1270] crA

[1271] TBDPSO^^. 'P V T

[1272]

[1273] O

[1274]

[0209] A mixture of (R)-7-((2-((tert-butyldiphenylsilyl)oxy)ethyl)sulfonyl)-2-(3-((S)-3-methoxy-2-methyl-3-oxopropyl)phenyl)-2,6,6-trimethylheptanoic acid (Intermediate 9H-1, 1.50 g, 2.16 mmol) and thionyl chloride (30 mL, 411 mmol) was stirred at 75 °C for 1 hour. The solvent was removed. The residue was dissolved in acetonitrile (10 mL), cooled to 0°C, and treated with trimethylsilyldiazomethane (4.3 mL, 8.63 mmol). The mixture was stirred at room temperature for 16 hours, and hydrobromic acid (4.50 eq, 1.3 mL, 9.71 mmol) was added. The reaction was stirred at room temperature for another 20 minutes, quenched with 30 mL of water, extracted with ethyl acetate (50 mL x 2). The combined organic extracts were washed with saturated sodium bicarbonate, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0-40% of ethyl acetate in petroleum ether) to give the title compound (1.26 g, 1.63 mmol, 75 %) as a white solid. MS (ESI): 793, 795 m / z [M+Na]+; purity: 90% (214 nm); retention time: 2.52 minutes (LC-MS Method 5).

[1275] Intermediate 24: 5-((2-Bromo-6-fluoro-4-((4-methoxybenzyl)oxy)-3-methylphenyl)thio)-2-fluorobenzonitrile

[1276]

[1277]

[0210] To a stirred solution of 5-((2-bromo-6-fluoro-4-hydroxy-3-methylphenyl)thio)-2-fluorobenzonitrile (2.60 g, 5.84 mmol) in N, N-dimethylformamide (20 mL) were added 4-methoxybenzylchloride (1.01 g, 6.42 mmol) and potassium carbonate (1.21 g, 8.76 mmol). The reaction mixture was stirred at 50 °C for 2 hours and diluted with ethyl acetate (100 mL). The 90

[1278] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1279] solution was washed with water, brine, dried over sodium anhydrous sulfate, filtered, and concentrated. The residue was purified by flash chromatography (80 g silica gel column, eluting with 0-15% ethyl acetate in petroleum ether) to give the title compound (2.70 g, 78 %) as a solid. 'H NMR (400 MHz, DMSO-<fc) 57.69 (dd, J= 5.8, 2.5 Hz, 1H), 7.51-7.39 (m, 3H), 7.39-7.31 (m, 2H), 7.02-6.93 (m, 2H), 5.14 (s, 2H), 3.77 (s, 3H), 2.29 (s, 3H) ppm.

[1280] Intermediate 24A: 5-((2-Bromo-6-fluoro-3-methyl-4-nitrophenyl)thio)-2-fluorobenzonitrile

[1281] Br

[1282]

[1283]

[0211] To a stirred solution of 2-fluoro-5-((4-methoxybenzyl)thio)benzonitrile (53.5 g, 196 mmol) in trifluoroacetic acid (200 mL) was added anisole (21.168 g, 196 mmol). The reaction was stirred at 50 °C for 16 hours under nitrogen. Triphenylphosphine (5 g) was added. The reaction mixture was concentrated. The residue was dissolved in N, N-dimethylformamide (300 mL), treated with 3-bromo-l,2-difluoro-4-methyl-5-nitrobenzene (44.42 g, 176 mmol) and potassium carbonate (54.10 g, 391 mmol). The reaction was stirred at room temperature for 3 hours under an argon atmosphere and diluted with ethyl acetate (500 mL). The solution was washed with water (200 mL x 2), brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (330 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (75.00 g, 76 %) as a yellow solid. MS (ESI): 385, 387 m / z [M+H]+; purity: 75% (214 nm); retention time: 2.14 minutes (LC-MS Method 2).

[1284] Intermediate 24B: 5-((4-Amino-2-bromo-6-fluoro-3-methylphenyl)thio)-2-fluorobenzonitrile

[1285] Br

[1286]

[1287]

[0212] To a stirred solution of 5-((2-bromo-6-fhioro-3-methyl-4-nitrophenyl)thio)-2-fluorobenzonitrile (Intermediate 24A, 50.00 g, 130 mmol) in acetic acid (500 mL) was added iron (21.75 g, 389 mmol). The reaction was stirred at room temperature for 4 hours. The reaction mixture was filtered. The filtrate was concentrated. The residue was dissolved in ethyl acetate (500 mL). The solution was washed with water, brine, dried over anhydrous sodium 91

[1288] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1289] sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (330 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (34.00 g, 71%) as a solid. MS (ESI): 355, 357 m / z [M+H]+; purity: 76% (214 nm); retention time: 2.07 minutes (LC-MS Method 2).

[1290] Intermediate 24C: 5-((2-Bromo-6-fluoro-4-hydroxy-3-methylphenyl)thio)-2-fluorobenzonitrile

[1291] Br

[1292]

[1293]

[0213] To a stirred solution of 5-(4-amino-2-bromo-6-fluoro-3-methyl-phenyl)sulfanyl-2-fluoro-benzonitrile (1.00 g, 2.82 mmol) in concentrated sulfuric acid (3.8 mL, 71.6 mmol) at 0 °C was added sodium nitrite (0.19 g, 2.82 mmol). The reaction mixture was stirred at 5 °C for 1 hour. The solution was added dropwise to a boiling mixture of toluene (4 mL) and water (40 mL) and stirred at 100 °C for 1 hour. The mixture was diluted with ethyl acetate (100 mL). The solution was washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-20% ethyl acetate in petroleum ether) to give the title compound (0.46 g, 46 %) as a solid. MS (ESI): 356, 358 m / z [M+H]+; purity: 91% (214 nm); retention time: 2.08 minutes (LC-MS Method 3).

[1294] Intermediate 25: 5-(2-Bromo-3,6-difluoro-4-methoxyphenoxy)-2-fluorobenzonitrile

[1295]

[1296]

[0214] To a suspension of 2-bromo-3,6-difluoro-4-methoxyphenol (Intermediate 25C, 1.5 g, crude, 2.5 mmol) in acetone (15 mL) was added (3-cyano-4-fluorophenyl)(2,4,6-trimethoxyphenyl)-l3-iodaneyl 4-methylbenzenesulfonate (1.46 g, 2.5 mmol) and potassium carbonate (1.04 g, 7.5 mmol). The reaction was stirred at 50 °C for 16 hours, quenched with water, and extracted with ethyl acetate (50 mL x 3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (0.66 g, 1.85 mmol, 73 % yield over two steps)

[1297] 92

[1298] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1299] as a yellow oil. MS (ESI): 358, 360 m / z [M+H]+; purity: 95% (214 nm); retention time: 2.10 minutes (LC-MS Method 6).

[1300] Intermediate 25A: 3-Bromo-l,2,5-trifluoro-4-((4-methoxybenzyl)oxy)benzene

[1301]

[1302]

[0215] To a suspension of 2-bromo-3,4,6-trifluorophenol (3.5 g, 15.5 mmol) and potassium carbonate (24.3 g, 31 mmol) in N, N-dimethylformamide (200 mL) was added paramethoxybenzyl chloride (2.7 g, 17.1 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours, quenched with water (300 mL), and extracted with ethyl acetate (300 mL x 3). The combined organic extracts were washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (1.4 g, 4 mmol, 26 %) as yellow oil. MS (ESI): 347, 349 m / z [M+H]+; purity: 76% (214 nm); retention time: 1.73 minutes (LC-MS Method 6).

[1303] Intermediate 25B: 3-bromo-l,4-difluoro-5-methoxy-2-((4-methoxybenzyl)oxy)benzene

[1304] Br

[1305]

[1306] F

[1307]

[0216] A mixture of 3-bromo-l,2,5-trifluoro-4-((4-methoxybenzyl)oxy)benzene (Intermediate 25 A, 1.9 g, 5.5 mmol) in pyridine / methanol / 30% sodium methoxide in methanol 3 / 2 / 1 (24 mL) was stirred at 50 °C for 16 hours. The mixture was quenched with water (100 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-10% ethyl acetate in petroleum ether) to give the title compound (0.9 g, 2.5 mmol, 46 % yield) as yellow oil. MS (ESI): 359, 361 m / z [M+H]+; purity: 95% (214 nm); retention time: 1.77 minutes (LC-MS Method 6).

[1308] Intermediate 25C: 2-Bromo-3,6-difluoro-4-methoxyphenol

[1309] Br

[1310]

[1311] 93

[1312] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1313]

[0217] A solution of 3-bromo-l,4-difluoro-5-methoxy-2-((4-methoxybenzyl)oxy)benzene (Intermediate 25B, 0.9 g, 2.5 mmol) in TFA (10 mL) and anisole (1 mL) was stirred at 50°C for 16 hours. The mixture was concentrated to give the crude title compound ( 1.5 g) as a yellow oil. MS (ESI): 239, 241 m / z [M+H]+; purity: 89% (214 nm); retention time: 1.84 minutes (LC-MS Method 6).

[1314] Preparation of Examples

[1315] Example 1. (2S)-3-[3-[(6R)-17-Amino-16-chloro-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12L6-thia-3, 4, 18, 27-tetrazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-1(26), 2(27),4,15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1316] , F

[1317] , /

[1318]

[1319] Methyl (S)-3-(3-((R)-2-(5-(5-((4-bromo-6-nitropyridin-3-yl)oxy)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate

[1320] O

[1321] O'

[1322]

[1323]

[0218] Step A: To a stirred solution of methyl (S)-3-(3-((R)-2-(5-(2-fluoro-5-hydroxyphenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2 -methylpropanoate (Intermediate 10, 1.11 g, 1.83 mmol) in N, N-dimethylformamide (15 mL) was added 4-bromo-5-fluoro-2-nitro-pyridine (Intermediate 4-1, 0.45 g, 2.04 mmol) and sodium bicarbonate (0.34 g, 4.07 mmol). The mixture was stirred at room temperature for 4 hours, then diluted with water (100 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (25 g silica gel column, eluting with 0-50% ethyl acetate in petroleum ether)

[1324] 94

[1325] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1326] to give the title compound (1.50 g,1.86 mmol, 92 %) as a yellow oil. MS (ESI): 806, 804 m / z [M+H]+; purity: 92% (214 nm); retention time: 2.03 minutes (LC-MS Method 2).

[1327] Methyl (S)-3-(3-((R)-2-(5-(5-((4-bromo-6-nitropyridin-3-yl)oxy)-2-fhiorophenyl)-l-methyl- 1 H-l,2,4-triazol-3-yl)-6, 6-dimethyl- 7-( vinylsulfonyl)heptan-2-yl )phenyl)-2-methylpropanoate

[1328]

[1329]

[0219] Step B: To a stirred and chilled (0 °C) solution of methyl (S)-3-(3-((R)-2-(5-(5-((4-bromo-6-nitropyridin-3-yl)oxy)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate (Step A product, 1.50 g, 1.86 mmol) in dichloromethane (11 mL) was added triethylamine (0.74 mL, 5.59 mmol) and methanesulfonyl chloride (0.17 mL, 2.24 mmol). The reaction mixture was stirred at room temperature for 2 hours. The volatiles were removed. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-50% ethyl acetate in petroleum ether) to give the title compound (1.30 g, 1.65 mmol, 89 %) as a yellow oil. MS (ESI): 788, 786 m / z [M+H]+; purity: 91% (214 nm); retention time: 2.13 minutes (LC-MS Method 2). Methyl ( S)-3-( 3-( (R)-2-( 5-( 5-( ( 6-amino-4-bromopyridin-3-yl)oxy)-2-fluorophenyl)-l-methyl-1 H-l,2,4-t riazol-3-yl)-6,6-dimethyl-7-( vmylsulfonyl)heptan-2-yl)phenyl)-2-methylpropanoate

[1330]

[1331]

[0220] Step C: To a stirred solution of Step B product (1.30 g, 1.65 mmol) in acetic acid (15 mL) was added iron (923 mg, 16.5 mmol). The reaction mixture was stirred at room temperature for 1 hour and filtered. The filtrate was adjusted with saturated aqueous sodium bicarbonate to pH ~ 8, extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with brine (80 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (1.10 g, 1.45 mmol, 88%) as a yellow oil. MS (ESI): 758, 756 m / z [M+H]+; purity: 91 % (214 nm); retention time: 1.92 minutes (LC-MS Method 2). Methyl ( S)-3-(3-( (R)-2-( 5-( 5-( ( 6-(bis( tert-butoxycarbonyl)amino)-4-bromopyridin-3-yl)oxy)-

[1332] 95

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[1334] 2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-6,6-dimethyl-7-(vinylsulfonyl)heptan-2-yl)phenyl)-2-methylpropanoate

[1335]

[1336]

[0221] Step D: To a mixture of Step C product (1.10 g, 1.45 mmol) in tetrahydrofuran (15 mL) was added di-tert-butyl dicarbonate (1.4 mL, 5.81 mmol), N, N-Diisopropylethylamine (1.0 mL, 5.81 mmol) and 4-dimethylaminopyridine (107 mg, 0.872 mmol). The mixture was stirred at room temperature overnight. The mixture was concentrated. The residue was purified by automated flash chromatography (80 g silica gel column, eluting with 0-50% ethyl acetate in petroleum ether) to give the title compound (1.01 g, 1.06 mmol, 73 %) as a light-yellow oil. MS (ESI): 958, 956 m / z [M+H]+; purity: 90% (254 nm); retention time: 2.31 minutes (LC-MS Method 2).

[1337] Methyl (2S)-3-[3-[( 6R, 13E)-17-[bis( tert-butoxycarbonyl)amino ]-25-fluoro-3, 6, 10,10-tetramethyl-12, 12-dioxo-21 -oxa- 12X6- thia- 3, 4,18,27-tetrazatetracyclo[ 20.3.1.12, 5.015,20]heptacosa-l ( 26), 2(27), 4, 13, 15, 17,19,22, 24-nonaen-6-yl (phenyl ]-2-methyl-propanoate

[1338]

[1339]

[0222] Step E: To a mixture of chloro[(tri-tert-butylphosphine)(2-aminobiphenyl-2-yl)palladium(II) (121 mg, 0.236 mmol), tri-tert-butylphosphine tetrafluoroborate (136 mg, 0.471 mmol) and N-cyclohexyl-N-methyl-cyclohexanamine (461 mg, 2.36 mmol) in toluene (10 mL) at 120 °C was added a solution of Step D product (1.13 g, 1.18 mmol) in toluene (4.0 mL) under an argon atmosphere. The mixture was stirred at 120 °C for 3 hours and concentrated. The residue was purified by automated flash chromatography (12.5 g silica gel column, eluting with 0-40% ethyl acetate in petroleum ether) to give the title compound (470 mg, 0.536 mmol, 46 %) as a white solid. MS (ESI): 876 m / z [M+H]+; purity: 56% (254 nm); retention time: 1.99 minutes (LC-MS Method 6).

[1340] 96

[1341] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1342] Methyl (2S)-3-[3-[(6R)-17-[bis(tert-butoxycarbonyl)amino]-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,16,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-1(26),2(27),4,15,17,19,22, 24-octaen-6-yl ] phenyl ] -2 -methyl -propanoate

[1343]

[1344]

[0223] Step F: To a stirred solution of Step E product (470 mg, 0.536 mmol) in ethanol (10 mL) was added 10% palladium on carbon (wetted with ca. 55% Water, 100 mg). The reaction mixture was stirred under hydrogen for 4 hours at 50 °C. The catalyst was removed by filtration and washed with ethanol (10 mL x 3). The filtrate was concentrated to give the title compound (440 mg, 0.501 mmol, 93 %) as a white solid. MS (ESI): 878 m / z [M+H]+; purity: 93% (254 nm); retention time: 2.28 minutes (LC-MS Method 2).

[1345] Methyl (2S)-3-l3-[(6R)-17-ammo-25-fluoro-3,6,10,10-tetramethyl- 12,12-dioxo-21 -oxa-1276-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl Jphenyl ]-2-methyl-propanoate

[1346]

[1347]

[0224] Step G: A mixture of Step F product (490 mg, 0.558 mmol) and hydrogen chloride in 1,4-dioxane (4.0 M, 5.0 mL, 20.0 mmol) was stirred at room temperature for 4 hours, then diluted with water (10 mL), adjusted the pH to ~ 7 with saturated sodium bicarbonate aqueous solution, and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (350 mg, 0.516 mmol, 93 %) as a light yellow solid. MS (ESI): 678 m / z [M+H]+; purity: 84% (254 nm); retention time: 1.74 minutes (LC-MS Method 5).

[1348] Methyl (2S)-3-[3-[(6R)-17-ammo-16-chloro-25-fhioro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4, 18,27-tetraz.atetracyclo[ 20.3.1.12,5.015, 20 Jheptacosa-

[1349] 97

[1350] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1351] 1(26), 2(27),- 4, 15, 17, 19, 22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoate

[1352]

[1353]

[0225] Step H: To a stirred solution of Step G product (90 mg, 0.133 mmol) in acetic acid (8 mL) was added N-chlorosuccinimide (53 mg, 0.398 mmol). The mixture was stirred at room temperature for 1 hour and another equivalent of N-chlorosuccinimide was added portion wise. After one hour the reaction mixture was quenched with water (20 mL), the pH adjusted to ~ 8 with saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate (15 mL x 3). The combined organic extracts were washed with saturated sodium bicarbonate aqueous solution (20 mL x 2), brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (4 g silica gel column, eluting with 0-100% ethyl acetate in petroleum ether) to give the title compound (66 mg, 0.0927 mmol, 69 %). MS (ESI): 713 m / z [M+H]+; purity: >99% (254 nm); retention time: 2.06 minutes (LC-MS Method 2).

[1354] Example 1: (2S)-3-[3-[(6R)-17-Amino-16-chloro-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4, 18,27-tetrazatetracyclo[ 20.3.1.12,5.015,20]heptacosa-1(26),2(27),4, 15,17,19,22,24-octaen-6-yl]phenyl ]-2-methyl-propanoic acid

[1355]

[1356]

[0226] Step I: To a stirred solution of Step H product (1.00 eq, 51 mg, 0.0716 mmol) in tetrahydrofuran (1 mL) was added a solution of lithium hydroxide (IM in water) (1 mL). The reaction was stirred at room temperature for 16 hours. The mixture was acidified with 1.0 M hydrochloric acid to pH ~ 6 and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase chromatography to afford the title compound (13 mg, 0.0179 mmol, 25 %) as a white solid. MS (ESI): 699 m / z [M+H]+; purity:

[1357] 98

[1358] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1359] >99% (254 nm); retention time: 2.09 minutes (LC-MS Method 7). ’f l NMR (400 MHz, CD3OD): 57.86 (s, 1H). 7.41-7.31 (m, 3H), 7.16 (t, J= 8.0 Hz, 1H), 7.05-6.98 (m, 3H), 3.85 (d, J= 2.0 Hz, 3H), 3.30-3.16 (m, 5H), 3.03-2.88 (m, 2H), 2.66-2.56 (m, 2H), 2.28-2.19 (m, 1H), 1.89-1.81 (m, 1H), 1.69 (s, 3H), 1.65-1.59 (m, 1H), 1.44-1.39 (m, 2H), 1.13-1.05 (m, 10H) ppm.

[1360] Example 2. (2R)-3-[3-[(6R)-18-Amino-16, 17-dichloro-19,25-difluoro-3, 6,10,10-tetramethyl-12,12,21-trioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1361]

[1362] Example 3 and 4: Diastereomers 1 and 2 of (2R)-3-[3-[(6R)-18-amino-16,17-dichloro-19,25-difluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1363]

[1364] Methyl ( 2R)-3-( 3-( ( 2R)-2-( 5-( 5-(aeetoxy( 3-(bis( tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)melhyl)-2-Jluorophenyl)-l-melhyl-lH-l,2,4-lriazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate

[1365]

[1366] 99

[1367] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1368]

[0227] Step A: To a stirred solution of (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate (Intermediate 11, 0.94 g, 1.14 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9, 0.48 g, 0.990 mmol) in pyridine (10 mL) were added 4A molecular sieves (0.7 g) at room temperature. The reaction mixture was stirred at 80 °C for 16 hours, cooled to room temperature, and quenched with IM hydrochloric acid (25 mL). The solution was extracted with ethyl acetate (90 mL). The organic extract was washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-80% ethyl acetate in petroleum ether) to give the title compound (0.44 g, 39.8 %) as a solid. MS (ESI): 1115, 1117 m / z [M+H]+; purity: 70% (254 nm); retention time: 2.95 minutes (LC-MS Method 4).

[1369] Methyl ( 2R)-3-( 3-( ( 2R)-2-( 5-( 5-( acetoxy! 3-(bis( tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)methyl)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-6,6-dimethyl- 7-( vinylsulfonyl)heptan-2-yl )phenyl)-2-methylpropanoate

[1370]

[1371]

[0228] Step B: To a stirred solution of Step A product (0.43 g, 0.385 mmol) in dichloromethane (10 mL) was added triethylamine (0.16 mL, 1.15 mmol) and methanesulfonyl chloride (0.045 mL, 0.577 mmol) at room temperature. The reaction mixture was stirred at room temperature for one hour and concentrated. The residue was purified by flash chromatography (25 g silica gel column, eluting with 0-50% ethyl acetate in petroleum ether) to give the title compound (0.34 g, 80 %) as a solid. MS (ESI): 1097, 1099 m / z [M+H]+; purity: 85% (254 nm); retention time: 3.15 minutes (LC-MS Method 4).

[1372] Methyl ( 2R)-3-[ 3-[ ( 6R, 13E)-21-acetoxy-18-[bis( tert-butoxycarbonyl)amino ]-16,l 7-dichloro-19,25-difluoro-3,6, 10,10-tetramethyl-12, 12-dioxo-12 / .6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,13,15,17,19,22,24-nonaen-6-yl Jphenyl ]-2-methyl-propanoate

[1373] 100

[1374] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1375]

[1376]

[0229] Step C: To a stirred solution of chloro[(tri-tert-butylphosphine)(2-aminobiphenyl-2-yl)palladium(II) (16 mg, 0.031 mmol), N-cyclohexyl-N-methyl-cyclohexanamine (121 mg, 0.62 mmol) and tri-tert-butylphosphine tetrafluoroborate (18 mg, 0.062 mmol) in toluene (110 mL) was added dropwise a solution of Step B product (340 mg, 0.31 mmol) in toluene (10 mL) over 1 h under an argon atmosphere. The reaction was stirred at 120 °C for 2 hours and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-60% ethyl acetate in petroleum ether) to give the title compound (140 mg, 45 %) as a white solid. MS (ESI): 1017 m / z [M+H]+; Peak A: purity: 40% (214 nm); retention time: 3.11 minutes; Peak B: purity: 41% (214 nm); retention time: 3.14 minutes; (Peak A and B are a diastereomer pair; LC-MS Method 4).

[1377] Methyl ( 2R)-3-[ 3-[ (6R)-21 -acetoxy-18-[bis( tert-butoxycarbonyl)amino 1-1, 17-dichloro- 19,25-difluoro-3,6,l(),10-tetramethyl-12,12-dioxo-12 / .6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl ]phenyl ]-2-methyl-propanoate

[1378]

[1379]

[0230] Step D: To a stirred solution of Step C product (125 mg, 0.123 mmol) in toluene (2 mL) was added p-toluenesulfonyl hydrazide (229 mg, 1.23 mmol). The reaction was stirred at 120 °C for 2 hours, cooled to room temperature, and filtered. The filter cake was washed with toluene (3 mL x 2). The filtrate was concentrated. The residue was diluted with ethyl acetate (20 mL), washed with 1.0 M hydrochloric acid (10 mL x 2), brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (125 mg, 99.8 %) as a solid. MS (ESI): 1019 m / z [M+H]+; purity: 92% (214 nm); retention time: 3.19 minutes (LC-MS Method 4).

[1380] 101

[1381] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1382] Methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-16,17-dichloro-19,25-difluoro-3,6,10,10-tetramethyl-12, 12-dioxo- 12X6-thia-3,4, 27-triazatetracyclo[ 20.3.1.12, 5.015, 20 ]heptacosa-1(26), 2(27), 4, 15,17,19,22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoate

[1383]

[1384]

[0231] Step E: To a stirred solution of Step D product (125 mg, 0.12 mmol) in dichloromethane (1 mL) was added 4M hydrogen chloride (1.0 mL, 3.80 mmol, in dioxane). The mixture was stirred at room temperature for 2 hours, quenched with water (20 mL), and extracted with ethyl acetate (30 mL x 3). The combined organic extract was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (20 g silica gel column, eluting with 0-40% ethyl acetate in petroleum ether) to give the title compound (65 mg, 65 %) as a solid. MS (ESI): 819 m / z [M+H]+; purity: 98% (254 nm); retention time: 2.77 minutes (LC-MS Method 4).

[1385] (2R)-3-[3-[(6R)- 18-amino-16, 17 -dichloro- 19, 25-difluoro-21 -hydroxy-3, 6, 10, 10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoic acid

[1386]

[1387] F

[1388]

[0232] Step F: To a stirred solution of Step E product (0.065 g, 0.079 mmol) in tetrahydrofiiran (0.70 mL) and methanol (0.24 mL) was added lithium hydroxide (0.24 mL, 0.238 mmol) (1 mol / L in water). The mixture was stirred at room temperature for 16 hours, acidified with 1.0 M hydrochloric acid to pH ~ 6, and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (55 mg, 90.8 %) as white solid. MS (ESI): 763 m / z [M+H]+; Peak A: purity: 40% (254 nm); retention time: 2.39 minutes; Peak B: Peak A: purity: 57% (254 nm); retention time: 2.45 minutes (Peak A and Peak B are two different diastereomers; LC-MS Method 4).

[1389] 102

[1390] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1391] Example 2: ( 2R )-3-[ 3-[ ( 6R )-18-Amino-16, 17-dichloro-l 9, 25-difluoro-3, 6,10,10-tetramethyl- 12,12,21-trioxo-12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26),2(27),4,15,17,19,22, 24-octaen-6-yl ] phenyl ]-2-methyl -propanoic acid

[1392]

[1393]

[0233] Step G: To a stirred solution of Step F product (35 mg, 0.046 mmol) in dimethylsulfoxide (1.5 mL) was added 2-iodoxybenzoic acid (57 mg, 0.092 mmol). The reaction was stirred at room temperature for 2 hours, quenched with water (5 mL), and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with aqueous sodium sulfite (5 mL), water (5 mL), brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (16 mg, 46.7 %) as a white solid. MS (ESI): 761 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.08 minutes (LC-MS Method 4). ’H NMR (400 MHz, CD3OD).

[1394] 58.29-8.25 (m, 1H), 8.10-8.07 (m. 1H), 7.58 (t, J = 9.2 Hz, 1H), 7.16 (t, J= 8.0 Hz, 1H), 7.06-6.98 (m, 3H), 3.88 (d. J= 2.4 Hz, 3H), 3.13-3.04 (m, 4H), 2.95-2.84 (m, 4H), 2.63-2.56 (m, 2H), 2.26-2.20 (m, 1H), 1.89-1.83 (m, 1H), 1.70 (s, 3H), 1.60-1.55 (m, 1H), 1.37-1.28 (m, 2H), 1.09-1.07 (m, 6H), 1.01 (s, 3H) ppm.

[1395] Example 3 and 4: Diastereomers 1 and 2 of(2R)-3-[3-[(6R)-18-amino-16,17-dichloro-19,25-difluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-12 / .6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl Jphenyl ]-2-methyl-propanoic acid

[1396]

[1397]

[0234] Step H: (2R)-3-|3-|(6R)-18-Amino-16,17-dichloro-19,25-difluoro-21-hydroxy- 3,6, 10, 10-tetramethyl-l 2, 12-dioxo- 12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6- 103

[1398] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1399] yljphenyl] -2 -methyl-propanoic acid (Step F product) (10 mg, 0.013 mmol) was purified by prep-HPLC to give the first eluent (2.0 mg, 20%, white solid, designated as Diastereomer 1, Example 3) and the second eluent (2.0 mg, 20%, white solid, designated as Diastereomer 2, Example 4)

[1400]

[0235] Example 3, Diastereomer 1: MS (ESI): 763 in / z [M+H]+; LC purity: 97% (214 nm); retention time: 1.99 minutes (LC-MS Method 2). ’H NMR (400 MHz, CD3OD). 58.04-7.99 (m, 1H), 7.29-7.24 (m, 1H), 7.18 (t, J= 7.6 Hz, 1H), 7.11-7.08 (m, 1H), 7.03-6.98 (m, 2H), 6.46 (s, 1H), 3.86 (d, J= 1.2 Hz, 3H), 3.13-2.82 (m, 8H), 2.66-2.57 (m, 2H), 2.35-2.28 (m, 1H), 1.87-1.78 (m, 2H), 1.72 (s, 3H), 1.49-1.35 (m, 3H), 1.22 (s, 3H), 1.11 (s, 3H), 1.08 (d, 7 = 6.8 Hz, 3H) ppm.

[1401]

[0236] Example 4, Diastereomer 2: MS (ESI): 763 m / z [M+H]+; LC purity: 98% (214 nm); retention time: 2.04 minutes (LC-MS Method 2).XH NMR (400 MHz, CD3OD). 58.05 (d, J = 5.2 Hz, 1H), 7.29-7.22 (m, 2H), 7.16-7.13 (m, 1H), 7.02-6.97 (m, 2H), 6.36 (s, 1H), 3.90 (d, 7 = 3.2 Hz, 3H), 3.13-3.05 (m, 2H), 2.99-2.82 (m, 6H), 2.64-2.56 (m, 2H), 2.35-2.30 (m, 1H), 2.18-2.09 (m, 1H), 1.71 (s, 3H), 1.50-1.36 (m, 3H), 1.30 (s, 3H), 1.16 (s, 3H), 1.08 (d, 7= 6.8 Hz, 3H), 1.00-0.96 (m, lH) ppm.

[1402] Example 5 and 6: Diastereomer 1 and 2 of (2R)-3-[3-[(6R)-18-Amino-16-chloro-17,19,25-trifluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1403] \

[1404] N

[1405] N, OH

[1406] , F

[1407] < ^NH.

[1408] r

[1409] F

[1410] \Cl

[1411] "s;

[1412]

[1413] 0 '0

[1414] Example 7 (2R)-3-[3-[(6R)-18-amino-16-chloro-17,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12X6-thia-3,4,27-triazatetracyclo

[1415] [20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1416] 104

[1417] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1418]

[1419] Example 8: (2R)-3-[3-[(6R)-18-Amino-17,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1420]

[1421] Methyl ( 2R)-3-(3-( ( 2R)-2-( 5-( 5-( acetoxy(3-(bis( tert-butoxycarbonyl)amino)-6-broino-5-chloro-2,4-difluorophenyl)niethyl)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)'phenyl)-2-methylpropanoate

[1422]

[1423]

[0237] Step A: To a stirred solution of (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1, 1.30 g, 1.91 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9, 0.92 g, 1.91 mmol) in pyridine (20 mb) were added 4A molecular sieves (1.30 g). The mixture was stirred at 80 °C for 16 hours, cooled to room temperature, and diluted with ethyl acetate (100 m ). The organic layer was washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (eluting with 50% ethyl acetate in petroleum ether to give the title compound (0.65 g, 0.591 mmol, 31 %) as a solid. MS (ESI): 1099, 1101 m / z [M+H]+; LC purity: 53% (214 nm); retention time: 2.28 minutes (LC-MS Method 5).

[1424] 105

[1425] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1426] Methyl ( 2R)-3-( 3-(( 2R)-2-( 5-( 5-( acetoxy( 3-(bis( tert-butoxycarbonyl )amino)-6-bromo-5-chloro-2,4-difluorophenyl)methyl)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-6,6-dimethyl-7-(vinylsulfonyl)heptan-2-yl)phenyl)-2-methylpropanoate

[1427] AcQ

[1428] ci

[1429]

[1430]

[0238] Step B: To a stirred solution of Step A product (1.00 eq, 0.65 g, 0.591 mmol) and triethylamine (0.25 mL, 1.77 mmol) in dichloromethane (10 mL) was added methanesulfonyl chloride (0.046 mL, 0.591 mmol). The mixture was stirred at room temperature for one-hour, diluted with dichloromethane (30 mL). The separated organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (eluting with 60% ethyl acetate in petroleum ether) to give the title compound (510 mg, 0.471 mmol, 80 %) as an oil. MS (ESI): 1081, 1083 m / z [M+H]+; LC purity: 71% (214 nm); retention time: 2.35 minutes (LC-MS Method 5).

[1431] Methyl (2R)-3-[3-[ ( 13E)-21 -acetoxy- 18-[bis( tert-butoxycarbonyl)amino ]-16-chloro- 17,19,25-trifluoro-3,6,10,l()-tetramethy -12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.()15,2()]heptacosa-l(26),2(27),4,13,15,17,19,22,24-nonaen-6-yl ]phenyl ]-2-methyl-propanoate

[1432] N

[1433] N

[1434]

[1435]

[0239] Step C: To a stirred solution of tBujP-HBF4 (13 mg, 0.0462 mmol), CyzNMe (0.099 mL, 0.462 mmol) and (tBu3P)Pd G2 (12 mg, 0.0231 mmol) in toluene (30 mL) was added dropwise over one hour at 120 °C a solution of the product of Step B (250 mg, 0.231 mmol) in toluene (10 mL). The mixture was stirred at 120 °C for 3 hours under Ar and concentrated. The residue was dissolved in ethyl acetate (100 mL), washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (eluting with 60% ethyl acetate in petroleum ether) to afford the title

[1436] 106

[1437] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1438] compound (160 mg, 0.160 mmol, 69 %) as a solid. MS (ESI): 1001 m / z [M+H]+; LC purity: 21% (214 nm); retention time: 2.35 minutes (LC-MS Method 5).

[1439] Methyl (2R)-3-[ 3-[ ( 6R)-21 -acetoxy-18-[bis( tert-butoxycarbonyl)amino ]-l 6-chloro-l 7,19,25-trifluoro-3, 6,10,10-tetramethyl-12, 12-dioxo-12X6-thia-3, 4, 27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl Iphenyl ]-2-methyl-propanoate

[1440]

[1441]

[0240] Step D: To a stirred solution of Step C product (320 mg, 0.320 mmol) in toluene (20 mL) was added p-toluenesulfonyl hydrazide (595 mg, 3.20 mmol). The mixture was stirred at 110 °C for 5 hours and concentrated. The residue was dissolved in ethyl acetate (50 mL). The solution was washed with water, brine, dried over sodium sulfate, filtered, and concentrated to afford the crude title compound (330 mg, 0.329 mmol, 100 % yield) as a solid. MS (ESI): 1003 m / z [M+H]+; LC purity: 11% (214 nm); retention time: 2.37 minutes (LC-MS Method 5). Methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-16-chloro-17,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4, 27 -triazatetracyclo [ 20.3.1.12, 5.015, 20 Jheptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1442]

[1443] And by-product, methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-17,19,25-trifluoro-3,6,10,10-tetramethyl-12, 12-dioxo-12X6-thia-3,4, 27-triazatetracyclo[ 20.3.1.12, 5.015, 20 Jheptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1444] 107

[1445] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1446]

[1447]

[0241] Step E: To a stirred solution of Step D product (330 mg, 0.329 mmol) in dichloromethane (2 mL) was added 4N hydrogen chloride in dioxane (2.0 mL, 8.0 mmol). The mixture was stirred at room temperature for 6 hours, the pH adjusted to ~ 8 with saturated aqueous sodium bicarbonate at 0 °C and extracted with dichloromethane (3 x 30 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (eluting with 70% ethyl acetate in petroleum ether) to afford the impure title compound (300 mg), which was further purified by prep-HPLC to afford the pure title compound (47 mg, 14 % yield, MS (ESI): 803 m / z [M+H]+; LC purity: 94% (214 nm); retention time: 2.69 minutes (LC-MS Method 4)) as a white solid, and a by-product, methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-17, 19,25-trifluoro-3,6, 10,10-tetramethyl- 12, 12 -dioxo- 12 6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (20 mg, 7.8 % yield, MS (ESI): 769 m / z [M+H]+; LC purity: 33+62% (214 nm, two diastereomers); retention time: 2.57 minutes (LC-MS Method 4)) as a white solid.

[1448] Example 5 and 6: Diastereomers 1 and 2 of (2R)-3-[3-[(6R)-18-Amino-16-chloro-17,19,25-trifluoro-21 -hydroxy-3, 6,10, 10-tetramethyl-12,12-dioxo-1276-thia-3, 4,27-triazatetracyclol 20.3.1.12,5.015,20 ]heptacosa-l( 26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl Jphenyl ]-2-methyl-propanoic acid

[1449] N

[1450] , OH

[1451] , F

[1452] £ F

[1453] \

[1454] -s;Cl

[1455]

[1456] 6''o

[1457]

[0242] Step F: To a stirred solution of methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-16-chloro- 17, 19,25 -trifluoro-3,6, 10, 10-tetramethyL 12,12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (First Step E product, 47 mg, 0.0373 mmol) in 108

[1458] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1459] tetrahydrofuran (1.5 mL) was added IM aqueous lithium hydroxide (1.5 mL, 1.5 mmol) and methanol (0.5 mL). The mixture was stirred at room temperature for 16 hours, acidified to pH ~ 5 with IN hydrochloric acid, and extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated to give the crude title compound (45 mg) as a solid. MS (ESI): 747 m / z [M+H]+; LC purity: 27+10% (214 nm, two diastereomers): retention time: 2.35 minutes (LC-MS Method 4).

[1460]

[0243] 15 mg of the above crude product was purified by prep-HPLC. The first eluent, Diastereomer f, was designated as Example 5 (4.5 mg, white solid); The second eluent, Diastereomer 2, was designated as Example 6 (9.0 mg, white solid).

[1461]

[0244] Example 5: MS (ESI): 747 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.95 minutes (LC-MS Method 5). ’H NMR (400 MHz, CD3OD) 57.99 (s, 1H), 7.33 (s, 1H), 7.26 (t, J= 9.2 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.09 (s, 1H), 7.02-6.97 (m, 2H), 6.43 (s, 1H), 3.86 (d, J= 1.2 Hz, 3H), 3.20-2.95 (m, 6H), 2.86 (s, 1H), 2.67-2.55 (m, 2H), 2.38-2.23 (m, 1H), 1.90-1.75 (m, 2H), 1.71 (s, 3H), 1.51-1.28 (m, 3H), 1.21 (s, 3H), 1.13-1.05 (m, 6H) ppm.

[1462]

[0245] Example 6: MS (ESI): 747 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.99 minutes (LC-MS Method 5). ‘H NMR (400 MHz, CD3OD, 5 8.04 (d. J= 6.0 Hz, 1H).

[1463] 7.31 - 7.18 (m, 2H), 7.14 - 7.08 (m, 2H), 6.98 (t, J = 7.6 Hz, 2H), 6.32 (s, 1H), 3.90 (d, J = 2.8 Hz, 3H), 3.10 - 3.01 (m, 2H), 2.99 - 2.90 (m, 3H), 2.89 - 2.79 (m, 2H), 2.68 - 2.56 (m, 2H), 2.38 - 2.27 (m. 1H), 2.17 - 2.10 (m, 1H), 1.70 (s, 3H), 1.57 - 1.35 (m, 3H), 1.31 (s, 3H), 1.15 (s, 3H), 1.12 (s, 1H), 1.09 (d, J = 6.8 Hz, 3H) ppm.

[1464] Example 7: ( 2R)-3-[ 3-[ ( 6R)-18-amino-16-chloro-l 7, 19, 25 -trifluoro- 3, 6, 10, 10-tetramethyl-12,12,21- trioxo- 12X6- thia- 3, 4, 27- triazatetracyclo [20.3.1.12,5.015,20 [heptacosa- 1 ( 26), 2(27), 4, 15, 17,19,22,24-octaen-6-yl [phenyl [-2-methyl-propanoic acid

[1465] \

[1466] N

[1467] N, F

[1468] NB

[1469] "^F

[1470]

[1471]

[0246] Step G: To a stirred solution of (2R)-3-[3-[(6R)-18-amino-16-chloro-17, 19,25-trifluoro-21 -hydroxy-3,6, 10,10-tetramethyl- 12, 12-dioxo- 12Z6-thia-3.4.27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl] -2-methyl-propanoic acid (Diastereomer mixture from Step F, 20 mg, 0.0198 mmol) in dimethyl sulfoxide (1 mL) was added 1-hydroxy-1,2-benziodoxol-3(1h)-one 1-oxide (2.00

[1472] 109

[1473] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1474] eq, 11 mg, 0.0396 mmol). The mixture was stirred at room temperature for 10 hours, diluted with ethyl acetate (50 mL), washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (4.6 mg, 0.00617 mmol, 31 % yield) as a solid. MS (ESI): 745 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.01 minutes (LC-MS Method 5).XH NMR (400 MHz, CD3OD) 58.25 (ddd, J = 8.8, 5.2, 2.4 Hz, 1H), 8.05 (dd, J = 6.8, 2.4 Hz, 1H), 7.56 (t, J = 9.2 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.04-6.68 (m, 3H), 3.87 (d, J = 2.0 Hz, 3H), 3.10-3.03 (m, 2H), 3.01-2.84 (m, 4H), 2.63-2.54 (m, 2H), 2.28-2.16 (m, 1H), 1.91-1.80 (m, 1H), 1.69 (s, 3H), 1.64-1.54 (m, 1H), 1.45-1.21 (m, 4H), 1.07 (d, J = 5.6 Hz, 6H), 1.01 (s, 3H) ppm.

[1475] ( 2R)-3-[3-[ ( 6R)-18-Amino-17, 19, 25 -trifluor o-21 -hydroxy-3, 6, 10, 10-tetramethyl-12,12-dioxo-12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1476]

[1477]

[0247] Step H: To a stirred solution of methyl (2R)-3-[3-[(6R)-21-acetoxy-18-amino-17,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (by-product from Step E, 20 mg, 0.0390 mmol) in tetrahydrofuran (1.5 mL) was added IM lithium hydroxide (1.5 mL, 1.5 mmol) and methanol (0.5 mL). The mixture was stirred at room temperature for 16 hours, acidified to pH ~ 5 with IN hydrochloric acid, and extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated to afford the crude title compound (35 mg, 100% yield) as solid. MS (ESI): 713 m / z [M+H]+; LC purity: 86% (214 nm, two diastereomers); retention time: 1.91 minutes (LC-MS Method 5).

[1478] Example 8: ( 2R)-3-[ 3-[ ( 6R)-18-Amino-l 7,19,25-trifluoro-3,6,10, 10-tetramethyl-12, 12,21-trioxo-12X6-thia-3,4,27-triaz.atetracyclo[ 20.3.1.12,5.015,20]heptacosa-1(26),2(27),4,15,17,19,22, 24-octaen-6-yl ] phenyl ]-2-methyl -propanoic acid

[1479] 110

[1480] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1481]

[1482]

[0248] Step I: To a stirred solution of Step H product (35 mg, 0.0363 mmol) in dimethyl sulfoxide (1 mL) was added 1-hydroxy-1,2-benziodoxol-3(1h)-one 1-oxide (20 mg, 0.0727 mmol). The mixture was stirred at room temperature for 5 hours, diluted with ethyl acetate (50 ml), washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (16 mg, 58 %) as a white solid. MS (ESI): 711 m / z [M+H]+; LC purity: 97% (214 nm); retention time: 1.97 minutes (LC-MS Method 5). ‘H NMR (400 MHz, CD3OD) 58.24 (ddd, 7= 8.8, 5.2, 2.4 Hz, 1H), 7.87 (dd, J = 6.8, 2.0 Hz, 1H), 7.57 (t, J = 10.4 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.08-6.93 (m, 4H), 3.83 (d, J = 2.0 Hz, 3H), 3.17-2.98 (m, 5H), 2.98-2.88 (m, 2H), 2.63-2.54 (m, 3H), 2.19 (td, J = 12.8, 3.2 Hz, 1H), 1.85 (td, J= 12.8, 5.2 Hz, 1H), 1.66 (s, 3H), 1.62-1.51 (m, 1H), 1.39 (td, J = 13.6, 5.2 Hz, 1H), 1.28-1.13 (m, 1H), 1.07 (d, J= 5.6 Hz, 6H), 1.03 (s, 3H) ppm.

[1483] Example 9: 6-(3,5-difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl-12,12,20-trioxo-1216,18,20Z4-trithia-3,4,16,26-tetrazatetracyclo[19.3.1.12,5.015,19]hexacosa-l(25),2(26),4,15(19),16,21,23-heptaen-17-amine

[1484]

[1485] Example 10: 6-(3,5-difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl-12,12,20,20-tetraoxo-12λ6,18,20λ6-trithia-3,4,16,26-tetrazatetracyclo[19.3.1.12,5.015,19]hexacosa-l(25),2(26),4,15(19),16,21,23-heptaen-17-amine

[1486]

[1487] 111

[1488] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1489] Tert-butyl (4-bromo-5-( ( 3-( 3-(2-( 3,5-difluorophenyl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)-l-methyl-lH-l,2,4-triazol-5-yl)-4-fluorophenyl)thio)thiazol-2-yl)carbamate

[1490]

[1491]

[0249] Step A: To a stirred solution of 2-(3,5-difluorophenyl)-7-((2-hydroxyethyl)sulfonyl)-N',2,6,6-tetramethylheptanehydrazide (Intermediate 9-3, 858 mg, 2.04 mmol) and methyl 5-((2-(bis(tert-butoxycarbonyl)amino)-4-bromothiazol-5-yl)thio)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-2, 1.31 g, 1.85 mmol) in pyridine (20 mL) were added 4A molecular sieves (2 g). The reaction was stirred at 50 °C for 16 hours under an argon atmosphere. The residue was filtered. The filter cake was washed with ethyl acetate (100 mL). The combined filtrate was concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-70% ethyl acetate in petroleum ether) to give the title compound (1.00 g, 1.20 mmol, 65 %). MS (ESI): 834, 832 m / z [M+H]+; LC purity: 90% (254 nm); retention time: 2.19 minutes (LC-MS Method 2).

[1492] Tert-butyl (4-bromo-5-( (3-(3-(2-( 3,5-difluorophenyl)-6, 6-dimethyl- 7-( vinylsulfonyl)heptan-2-yl)-1-methyl-1H-1,2,4-triazol-5-yl)-4-fluorophenyl)thio)thiazol-2-yl)carbamate

[1493]

[1494]

[0250] Step B: To a stirred solution of Step A product ( 1.00 g, 1.20 mmol) in dichloromethane (10 mL) were added triethylamine (1.7 mL, 12.0 mmol) and methanesulfonyl chloride (0.19 mL, 2.40 mmol). The reaction mixture was stirred at room temperature for 4 hours, quenched with saturated sodium bicarbonate (40 mL), and extracted with ethyl acetate (50 mL x 2). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (40 g silica gel column, eluting with 0-60% ethyl acetate in petroleum ether) to give the title compound (960

[1495] 112

[1496] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1497] mg, 1.22 mmol, 96 %). MS (ESI): 816, 814 m / z [M+H]+; LC purity: 70% (254 nm); retention time: 2.27 minutes (LC-MS Method 2).

[1498] Tert-butyl (4-bromo-5-((3-(3-(2-(3,5-difluorophenyl)-6,6-dimethyl-7-(vinylsulfonyl)heptan-2-yl)-l-methyl-lH-l,2,4-triazol-5-yl)-4-fluorophenyl)thio)thiazol-2-yl)(tertbutoxycarbonyl)carbamate

[1499]

[1500]

[0251] Step C: To a stirred solution of Step B product (520 mg, 0.638 mmol) in toluene (5 mL) were added 4-dimethylaminopyridine (39 mg, 0.319 mmol) and di-tert-butyl dicarbonate (0.30 mL, 1.28 mmol) at room temperature. The reaction was stirred at 80 °C for 40 minutes then concentrated. The residue was purified by flash chromatography (25 g silica gel column, eluting with 0-50 % ethyl acetate in petroleum ether) to give the title compound (300 mg, 0.328 mmol, 51 %). MS (ESI): 916, 914 m / z [M+H]+; LC purity: 93% (254 nm); retention time: 2.51 minutes (LC-MS Method 2).

[1501] Tert-butyl N-tert-butoxycarbonyl-N-[(13E)-6-(3,5-difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-12λ6,18,20-trithia-3,4,16,26-tetrazatetracyclo[19.3.1.1²,⁵.0¹⁵,¹⁹]hexacosa-1(25),2(26),4,13,15(19),16,21,23-octaen-17-yl]carbamate

[1502]

[1503]

[0252] Step D: To a stirred solution of chloro[(tri-tert-butylphosphine)(2-aminobiphenyl-2-yl)palladium(II) (34 mg, 0.0656 mmol), tri-tert-butylphosphine tetrafluoroborate (38 mg, 0.131 mmol) and N-cyclohexyl-N-methyl-cyclohexanamine (256 mg, 1.31 mmol) in toluene (50 mL) at 120 °C was added dropwise a solution of Step C product (300 mg, 0.328 mmol) in toluene (3 mL) under an argon atmosphere. The reaction was stirred at 120 °C for 2 hours. The solvent was removed. The residue was purified by flash chromatography (330 g silica gel column,

[1504] 113

[1505] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1506] eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (170 mg, 0.204 mmol, 62 %). MS (ESI): 834 m / z [M+H]+; LC purity: 94% (214 nm); retention time: 2.49 minutes (LC-MS Method 2).

[1507] Tert-butyl N-tert-butoxycarbonyl-N-[6-(3,5-difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl- 12,12-dioxo-1276,18,20-trithia-3,4,16,26-tetrazatetracyclo[19.3.1.12,5.015,19]hexacosa- 1(25), 2(26), 4, 15(19), 16,21, 23 -heptaen- 17-yl ] carbamate

[1508]

[1509]

[0253] Step E: To a stirred solution of Step D product ( 150 mg, 0.180 mmol) in toluene (5 mL) was added p-toluenesulfonyl hydrazide (335 mg, 1.80 mmol). The reaction mixture was stirred at 120 °C for 2 hours under an argon atmosphere, cooled to room temperature, quenched with water (40 mL), and extracted with ethyl acetate (50 mL x 2). The combined organic extracts were concentrated. The residue was purified by flash chromatography (330 g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (60 mg, 0.0718 mmol, 39.9 %). MS (ESI): 836 m / z [M+H]+; LC purity: 90% (214 nm); retention time: 2.51 minutes (LC-MS Method 2).

[1510] Mixture of di-tert-butyl (Z)-(13-( 3,5-difluorophenyl)-26-fluoro- 11, 9, 9,13-tetramethyl-3, 7, 7-trioxido-HH-3, 7-dithia-4( 5,4)-thiazola-l ( 5,3)-triazola-2( l,3)-benzenacyclotridecaphane-42-yl)iminodicarbonate and di-tert-butyl (Z)-(l 3-(3,5-difluorophenyl)-26-fluoro-l 1,9,9,13-letramethyl-3, 3, 7, 7-letraoxido-llH-3, 7-dithia-4( 5,4)-thiazola-l (5,3 )-triazola-2(l,3 )-benzenacyclotridecaphane-42-yl)iminodicarbonate

[1511]

[1512]

[0254] Step F: To a stirred solution of Step E product (60 mg, 0.0718 mmol) in methanol (3 mL) was added a solution of ammonium molybdate tetrahydrate (53 mg, 0.0431 mmol) in hydrogen peroxide (70% in water, 0.20 mL). The mixture was stirred at room temperature for 114

[1513] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1514] 1 hour, quenched with water (30 mL), and extracted with ethyl acetate (40 mL x 2). The combined organic phase was washed with saturated sodium thiosulfate (20 mL), brine, dried over sodium sulfate, filtered, and concentrated to give the crude title mixture (60 mg), which was used in the next step without further purification. MS (ESI): 852 m / z [M+H]+; LC purity: 54% (214 nm); retention time: 2.28 minutes (LC-MS Method 2); MS (ESI): 868 m / z [M+H]+; LC purity: 24% (214 nm); retention time: 2.36 minutes (LC-MS Method 2).

[1515] Example 9: 6-( 3, 5-difluorophenyl)-24-fluoro-3, 6,10,10-tetramethyl-12, 12,20-trioxo-12 / 6, 18,20 / 4-trithia-3,4,16,26-tetrazatetracyclo[ 19.3.1.12,5.015,19 Jhexacosa-l(25),2(26),4,15(19),16,21,23-heptaen-17-amine

[1516]

[1517] Example 10: 6-( 3,5-difluorophenyl)-24-fluoro-3,6, 10,10-tetramethyl-l 2, 12, 20, 20-tetraoxo-12 / 6, 18,20 / 6-trithia-3,4, 16,26-tetrazatetracyclo[ 19.3.1.12, 5.015,19 Jhexacosa-1(25),2(26),4, 15(19),! 6,21,23-heptaen-l 7-amine

[1518]

[1519]

[0255] Step G: To a stirred solution of Step F mixture (60 mg, 0.0704 mmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.2 mL). The reaction was stirred at room temperature for 2 hours and concentrated. The residue was treated with saturated sodium bicarbonate (20 mL) and extracted with ethyl acetate (40 mL x 2). The combined organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated. The crude was purified by prep-HPLC to give the first eluent, designated as Example 9 (26 mg, 0.0397 mmol, 55 %) and the second eluent, designated as Example 10 (7.1 mg, 0.0106 mmol, 15 %).

[1520]

[0256] Example 9: MS (ESI): 652 m / z [M+H]+; LC purity: 99% (254 nm); retention time: 1.94 minutes (LC-MS Method 2).1H NMR (400 MHz, CD₃OD) 5 8.26-8.06 (m, 2H), 7.61 (t, J =

[1521] 115

[1522] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1523] 9.2 Hz, 1H), 6.84 (d, J = 7.4 Hz, 2H), 6.77-6.67 (m, 1H), 3.80 (s, 3H), 3.77-3.65 (m, 1H), 3.51-3.36 (m, 2H), 3.12-2.84 (m, 3H), 2.48-1.83 (m, 3H), 1.72 (s, 3H), 1.52-1.27 (m, 2H), 1.24-0.97 (m, 7H) ppm.

[1524] Example 10: MS (ESI): 668 m / z [M+H]+; LC purity: 99% (254 nm); retention time: 2.05 minutes (LC-MS Method 2). H NMR (400 MHz, CD3OD) 58.40-8.30 (m, 1H), 8.21-8.10 (m, 1H), 7.58 (t, J = 9.2 Hz, 1H), 6.91-6.81 (m, 2H), 6.80-6.70 (m, 1H), 3.86 (s, 3H), 3.43-3.33 (m, 2H), 3.27-3.20 (m, 2H), 3.02 (s, 2H), 2.36-2.24 (m, 1H), 2.04-1.88 (m, 1H), 1.73 (s, 3H), 1.64-1.32 (m, 3H), 1.20-1.10 (m, 7H) ppm.

[1525] Example 11: (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12L6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1526]

[1527]

[0257] Exchanging 2-(3,5-difluorophenyl)-7-((2-hydroxyethyl)sulfonyl)-N', 2,6,6-tetramethylheptanehydrazide (Intermediate 9-2) with methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9, 2.50 g, 5.16 mmol) and methyl 5-((2-(bis(tert-butoxycarbonyl)amino)-4-bromothiazol-5-yl)thio)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-2) with methyl 5-(4-amino-2-bromo-6-fluorophenoxy)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-3, 1.93 g, 5.16 mmol), the reaction procedure sequence (Steps A, B, C, D described for Example 9 and 10, followed by Steps F, G, H and I described for Example 1) was used to prepare the title compound (7.0 mg, white solid). MS (ESI): 749 m / z [M+H]+; LC purity: >99% (254 nm); retention time: 2.09 minutes (LC-MS Method 5).1H NMR (400 MHz, CD₃OD) δ 7.38 (dd, J = 5.6, 3.2 Hz, 1H), 7.33 (t, J = 9.2 Hz, 1H), 7.26-7.19 (m, 1H), 7.16 (t, 7 = 8.0 Hz, 1H), 7. O5 (s, 1H), 7. OO (d, 7 = 7.2 Hz, 2H), 3.87 (d, 7 = 2.0 Hz, 3H), 3.26-3.15 (m, 4H), 3.14-3.08 (m, 1H), 3.03 (d, 7= 13.6 Hz, 1H), 2.97-2.91 (m, 1H), 2.66-2.53 (m, 2H), 2.22 (td, 7= 13.2, 4.4 Hz, 1H), 1.85 (td, 7= 13.2, 4.4 Hz, 1H), 1.76-1.61 (m, 4H). 1.48-1.26 (m, 2H), 1.20-1.15 (m, 1H), 1.12 (s, 6H), 1.07 (d, 7= 6.8 Hz, 3H) ppm.

[1528] 116

[1529] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1530] Example 12: Eluent 1, Diastereomer 1 of (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-12i6-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1531]

[1532] Example 13, Eluent 2, Diastereomer 2 of (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1533]

[1534] Example 14: (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12k6-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1535]

[1536]

[0258] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1) with (4-(bis(tert-butoxycarbonyl)amino)-2-bromo-6-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (3.9 g, 3.71 mmol), the reaction procedure sequence (Steps A, B, C, D, E described for Examples 5, 6, 7 and 8,

[1537] 117

[1538] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1539] followed by Steps H described for Example 1, and Steps F and G described for Examples 5, 6, 7 and 8) was used to prepare the title compounds.

[1540]

[0259] Example 12: Eluent 1, Diastereomer 1 of (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl] -2 -methyl-propanoic acid (4.9 mg, white solid). MS (ESI): 763 m / z [M+H]+; LC purity: >99% (254 nm); retention time: 1.95 minutes (LC-MS Method 5).JH NMR (500 MHz, CD3OD) 57.96 (s, 1H), 7.41-7.20 (m, 2H), 7.17 (t, J = 7.5 Hz, 1H), 7.09 (s, 1H), 7.00 (dd, 7 = 12.5, 7.9 Hz, 2H), 6.38 (s, 1H), 3.86 (s, 3H), 3.40-3.33 (m, 1H), 3.28-3.20 (m, 1H), 3.17-3.10 (m, 1H), 3.03 (s, 2H), 3.01-2.95 (m, 2H), 2.67-2.56 (m, 2H), 2.36-2.30 (m, 1H), 1.86-1.77 (m, 2H), 1.71 (s, 3H), 1.43-1.37 (m, 3H), 1.20 (s, 3H), 1.15-1.06 (m, 6H) ppm.

[1541]

[0260] Example 13, Eluent 2, Diastereomer 2 of (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl] -2 -methyl-propanoic acid (5.2 mg, white solid). MS (ESI): 763 m / z [M+H]+; LC purity: >99% (254 nm); retention time: 1.98 minutes (LC-MS Method 5). ’H NMR (500 MHz, CD3OD) 57.96 (s, 1H), 7.41-7.20 (m, 2H), 7.17 (t, 7 = 7.5 Hz, 1H), 7.09 (s, 1H), 7.00 (dd, 7 = 12.5, 8.0 Hz, 2H), 6.38 (s, 1H), 3.86 (s, 3H), 3.40-3.33 (m, 1H), 3.28-3.20 (m, 1H), 3.17-3.10 (m, 1H), 3.03 (s, 2H), 3.02-2.96 (m, 2H), 2.67-2.56 (m, 2H), 2.36-2.30 (m, 1H), 1.87-1.80 (m, 2H), 1.71 (s, 3H), 1.44-1.35 (m, 3H), 1.20 (s, 3H), 1.15-1.06 (m, 6H) ppm.

[1542]

[0261] Example 14: (2R)-3-[3-[(6R)-17-amino-16,18-dichloro-19,25-difhioro-3,6,10,10-tetramethyl-12,12,21-trioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (6.0 mg, white solid). MS (ESI): 761 m / z [M+H]+; LC purity: >99% (254 nm); retention time: 2.01 minutes (LC-MS Method 5).1H NMR (500 MHz, CD3OD) 5 8.22-8.18 (m, 1H), 8.05 (dd, 7 = 6.5, 2.0 Hz, 1H), 7.55 (t, 7= 9.0 Hz, 1H), 7.15 (t, 7= 7.5 Hz, 1H), 7.03 (d, 7= 8.0 Hz, 2H), 6.99 (d, J = 7.5 Hz, 1H), 3.86 (d, 7 = 2.0 Hz, 3H), 3.39-3.35 (m, 1H), 3.28 (s, 1H), 3.23-3.11 (m, 3H), 2.92-2.86 (m, 2H), 2.62-2.57 (m, 2H), 2.25-2.19 (m, 1H), 1.88-1.82 (m, 1H), 1.68 (s, 3H), 1.64-1.57 (m, 1H), 1.40-1.35 (td, 7 = 8.0, 4.0 Hz, 1H), 1.28 (td, 7= 8.0, 4.5 Hz, 1H), 1.08-1.05 (m, 6H), 1.02 (d, 7= 10.5 Hz, 4H) ppm.

[1543] Example 15. (2S)-3-[3-[(6R)-17-amino-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12Z6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1544] 118

[1545] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1546]

[1547]

[0262] Methyl (2S)-3-[3-[(6R)-17-amino-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-I2X6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step G product of Example 1) was subjected to the reaction conditions described for Step I of Example 1 to afford the title compound as a white solid (2.1 mg). MS (ESI): 664 m / z [M+H]+; LC purity: 98% (254 nm); retention time: 1.67 minutes (LC-MS Method 2). ’HNMR (400 MHz, CDsOD) 8 7.53-7.49 (m, 2H), 7.43 (t, J= 10.0 Hz, 1H), 7.37 (s, 1H), 7.15 (t, J= 8.0 Hz, 1H), 7.04-6.95 (m, 4H), 3.86 (d, 7= 2.8 Hz, 3H), 3.50-3.46 (m, 2H), 3.26-3.20 (m, 1H), 3.07-2.90 (m, 3H), 2.64-2.55 (m, 2H), 2.29-2.24 (m, 1H), 1.83 (t, J= 11.2 Hz, 1H), 1.66 (s, 3H), 1.60-1.52 (m. 2H), 1.46-1.28 (m, 2H), 1.14 (s, 3H), 1.06-1.00 (m, 7H) ppm.

[1548] Example 16: 18,19,25-trifluoro-6,10,10-trimethyl-3-[2-(methylamino)ethyl]-12,12-dioxo-6-phenyl-21-oxa-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-amine

[1549]

[1550] 4-(3-(1-(2-(Benzyloxy)ethyl)-3-((2-((tert-butyldiphenylsilyl)oxy)ethyl)sulfonyl)-6,6-dimethyl-2-phenylheptan-2-yl)-1H-1,2,4-triazol-5-yl)-4-fluorophenoxy)-5-bromo-2,3-difluoroaniline

[1551]

[1552]

[0263] Step A: To a stirred solution of N'-(2-(benzyloxy)ethyl)-7-((2-((tert-butyldiphenylsilyl)oxy)ethyl)sulfonyl)-2,6,6-trimethyl-2-phenylheptanehydrazide (Intermediate 9-1, 3.44 g, 4.63 mmol) and methyl 5-(4-amino-6-bromo-2,3-difluorophenoxy)- 119

[1553] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1554] 2-fluorobenzimidothioate hydroiodide (Intermediate 11-6, 3.70 g, 4.63 mmol) in pyridine (30 ml) were added 4A molecular sieves (4.00 g). The mixture was stirred at 50 °C for 2 days, dissolved in ethyl acetate (100 ml), washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (eluting with 50% ethyl acetate in petroleum ether) to give the title compound (3.00 g, 2.81 mmol, 61 %) as a pale yellow solid. MS (ESI): 1067, 1069 m / z [M+H]+; LC purity: 83% (254 nm); retention time: 2.67 minutes (LC-MS Method 2).

[1555] 2-(6-(5-(5-(4-Amino-6-bromo-2,3-difluorophenoxy)-2-fluorophenyl)-1-(2-benzyloxy)ethyl)-1H-1,2,4-triazol-3-yl)-2,2-dimethyl-6-phenylheptyl)sulfonyl)ethan-1-ol

[1556]

[1557]

[0264] Step B: To a stirred solution of Step A product (3.00 g, 2.81 mmol) in tetrahydrofuran (50 ml) was added 1M tetrabutylammonium fluoride in tetrahydrofuran (5.6 ml, 5.6 mmol). The mixture was stirred at room temperature for 2 hours and diluted with ethyl acetate (200 ml). The solution was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography eluting with 50% ethyl acetate in petroleum ether to give the title compound (1.40 g, 1.69 mmol, 60 %) as a pale yellow solid. MS (ESI): 829, 831 m / z [M+H]+; LC purity: 98% (214 nm); retention time: 2.09 minutes (LC-MS Method 5).

[1558] Tert-Butyl N-tert-butoxycarbonyl-N-[18, 19, 25-trifluoro-3-(2-hydroxyethyl)-6, 10,10-trimethyl-12,12-dioxo-6-phenyl-21-oxa-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl ] carbamate

[1559]

[1560]

[0265] Step C: Exchanging methyl (S)-3-(3-((R)-2-(5-(5-((4-bromo-6-nitropyridin-3-yl)oxy)- 2-fluorophenyl)- 1 -methyl- 1 H- 1,2,4-triazol-3 -yl)-7 -((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate (Step A product of Example 1) with 2-((6- 120

[1561] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1562] (5-(5-(4-Amino-6-bromo-2,3-difluorophenoxy)-2-fluorophenyl)-l-(2-(benzyloxy)ethyl)-lH-l,2,4-triazol-3-yl)-2,2-dimethyl-6-phenylheptyl)sulfonyl)ethan-l-ol (Step B product, 1.40 g, 1.69 mmol), the reaction procedure sequence (Steps B, D, E and F described for Example 1) was used to prepare the title compound (510 mg, 0.605 mmol) as a pale yellow solid. MS (ESI): 843 m / z [M+H]+; LC purity: 78% (254 nm); retention time: 2.25 minutes (LC-MS Method 5). Tert-butyl N-tert-butoxycarbonyl-N-[ 18,19,25-trifluoro-6, 10, 10-trimethyl-l 2, 12-dioxo-3-( 2-oxoethyl)-6-phenyl-21-oxa-121.6-thia-3,4,27-triazatetracyclo[2().3.1.12,5.015,20]heptacosa-1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-17-yl ] carbamate

[1563]

[1564]

[0266] Step D: To a stirred solution of Step C product (510 mg, 0.605 mmol) in dimethyl sulfoxide (5 ml) was added l,l,l-tris(acetyloxy)-l,l-dihydro-l,2-benzodioxol-3-(lh)-one (513 mg, 1.21 mmol). The mixture was stirred at room temperature for 2 hours and concentrated. The residue was dissolved in ethyl acetate (50 ml). The solution was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography (eluting with 50%Ethyl acetate in petroleum ether) to give the title compound (400 mg, 0.166 mmol, 28 %) as a yellow solid. MS (ESI): 841 m / z [M+H]+; LC purity: 35% (254 nm); retention time: 2.30 minutes (LC-MS Method 2).

[1565] Tert-butyl N-tert-butoxycarbonyl-N-[18,19,25-trifluoro-6,10,10-trimethyl-3-[2- ( methylamino)ethyl ]-12, 12-dioxo-6-phenyl-2 l-oxa-12 6-thia-3,4, 27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl Jcarbamate

[1566]

[1567]

[0267] Step E: A mixture of step D product (200 mg, 0.238 mmol) and methanamine (2M in tetrahydrofuran) (0.18 ml, 0.357 mmol) in dichloroethane (10 ml) was stirred for one hour, then treated with sodium triacetoxyborohydride (101 mg, 0.476 mmol). The mixture was stirred 121

[1568] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1569] at room temperature overnight and concentrated. The residue was dissolved in ethyl acetate (50 ml). The solution was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography (eluting with 50% ethyl acetate in petroleum ether) to give the title compound (30 mg, 0.0350 mmol, 15 %) as a yellow solid. MS (ESI): 856 m / z [M+H]+; LC purity: 70% (254 nm); retention time: 1.75 minutes (LC-MS Method 5).

[1570] Example 16: 18,19,25-trifluoro-6,l(),10-trimethyl-3-[2-(methylamino)ethyl]-12,12-dioxo-6-phenyl-21 -oxa- 12X6-thia-3,4,27-triazatetracyclo[ 20.3.1.12, 5.015, 20 ]heptacosa-1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-17-amine

[1571]

[1572]

[0268] Step F: To a stirred solution of Step E product (23 mg, 0.0269 mmol) in dichloromethane (5 ml) was added 4N HC1 in dioxane (5.0 ml, 20.0 mmol). The mixture was stirred at room temperature for 3 hours, basified with saturated aqueous sodium bicarbonate to pH ~ 8, and extracted with dichloromethane (30 ml x 3). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The crude product was purified by prep-HPLC to give the title compound (1.8 mg, 0.00274 mmol, 10 %). MS (ESI): 656 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.66 minutes (LC-MS Method 2). ‘H NMR (400 MHz, CDsOD) 6 7.39-7.32 (m, 2H), 7.29-7.23 (m, 2H), 7.22-7.13 (m, 4H), 6.63 (dd, 7= 8.8, 2.0 Hz, 1H), 4.36-4.21 (m, 2H), 3.22-3.15 (m, 3H), 3.14-3.09 (m, 1H), 3.03-2.95 (m, 3H), 2.82 (d, 7= 13.6 Hz, 1H), 2.42 (s, 3H), 2.25-2.14 (m, 1H), 1.87 (td, 7= 12.8, 4.4 Hz, 1H), 1.75-1.56 (m, 5H). 1.49-1.39 (m, 1H), 1.32 (s, 1H), 1.13 (s, 3H), 1.08 (s, 3H) ppm.

[1573] Example 17: 16-Chloro-l 8,19,25-trifluoro-6,10, 10-trimethyl-3-[ 2-( methylamino)ethyl]- 12,12-dioxo-6-phenyl-21-oxa-12X6-thla-3,4,27-triazatetracyclo[20.3.1.12, 5.015,20 ]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen- 17-amine

[1574] 122

[1575] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1576]

[1577] 2-((6-(5-(5-(4-Amino-2-bromo-3-chloro-5,6-difluorophenoxy)-2-fluorophenyl)-l-(2- (benzyloxy)ethyl)-lH-l,2,4-triazol-3-yl)-2,2-dimethyl-6-phenylheptyl)sulfonyl)ethan-l-ol

[1578]

[1579]

[0269] Step A: To a stirred solution of 2-((6-(5-(5-(4-amino-6-bromo-2,3-difluorophenoxy)-2-fluorophenyl)-l-(2-(benzyloxy)ethyl)-lH-l,2,4-triazol-3-yl)-2,2-dimethyl-6-phenylhexyl)sulfonyl)ethan-l-ol (Step B product of Example 16, 0.98 g, 1.18 mmol) in acetic acid (10 mL) was added N-chlorosuccinimide (189 mg, 1.42 mmol). The mixture was stirred at room temperature for 5 hours and concentrated. The residue was dissolved in ethyl acetate (50 mL), washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (eluting 50% ethyl acetate in petroleum ether) to give the title compound (750 mg, 73 %) as a solid. MS (ESI): 865. 863 m / z [M+H]+; LC purity: 96% (254 nm); retention time: 2.14 minutes (LC-MS Method 5).

[1580] Tert-butyl N-[3-(2-benzyloxyethyl)-16-chloro-18,19,25-trifluoro-6,10,10-trimethyl-12,12-dioxo-6-phenyl-21-oxa-l 276-thia-3,4, 27-triazatetracyclo[20.3.1.12, 5.015, 20 Iheptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl]-N-tert-butoxycarbonyl-carbamate

[1581]

[1582]

[0270] Step B: Exchanging Methyl (S)-3-(3-((R)-2-(5-(5-((4-bromo-6-nitropyridin-3-yl)oxy)-2-fluorophenyl)-l-methyl-lH-l,2,4-triazol-3-yl)-7-((2-hydroxyethyl)sulfonyl)-6,6-dimethylheptan-2-yl)phenyl)-2-methylpropanoate (Step A product of Example 1) with 2-((6-(5-(5-(4-amino-2-bromo-3-chloro-5,6-difluorophenoxy)-2-fluorophenyl)-l-(2- (benzyloxy)ethyl)-lH-l,2,4-triazol-3-yl)-2,2-dimethyl-6-phenylheptyl)sulfonyl)ethan-l-ol 123

[1583] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1584] (Step A product, 750 mg, 0.88 mmol), the reaction procedure sequence (Steps B, D, E and F described for Example 1) was followed to prepare the title compound (270 mg) as a solid. MS (ESI): 967 m / z [M+H]+; LC purity: 93% (254 nm); retention time: 2.53 minutes (LC-MS Method 5).

[1585] 2-(17-Amino-16-chloro-18,19,25-trifluoro-6,10,10-trimethyl-12,12-dioxo-6-phenyl-21-oxa- 12X6-thia-3,4, 27-triazatetracyclo[ 20.3.1.12,5.015,20 Jheptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-3-yl)ethanol

[1586]

[1587]

[0271] Step C: To a stirred solution of Step B product (270 mg, 0.28 mmol) in dichloromethane (27 mL) was added boron tribromide (2.7 mL, 28.6 mmol) at 0 °C. The mixture was stirred at 0 °C for 0.5 hours, treated with methanol (27 mL), and stirred at 0 °C for another 0.5 hours. The reaction was concentrated. The residue was dissolved in ethyl acetate (100 mL), washed with water, brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography (eluting with 70% ethyl acetate in petroleum ether) to give the title compound (75 mg, 39 %) as a solid. MS (ESI): 677 m / z [M+H]+; LC purity: 98% (254 nm); retention time: 2.04 minutes (LC-MS Method 5).

[1588] 2-(l 7-Amino-16-chloro-18, 19,25-trifluoro-6,10, 10-trimethyl-12, 12-dioxo-6-phenyl-21 -oxa- 12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-3-yl)ethyl tert-butyl carbonate

[1589]

[1590]

[0272] Step D: To a stirred solution of Step B product (1.00 eq, 70 mg, 0.103 mmol) in 5 mL of toluene was added Boc anhydride (10 mL, 42.7 mmol). The mixture was stirred at reflux for 16 hours and concentrated. The residue was dissolved in ethyl acetate (100 mL). The solution was washed with water, brine, dried over sodium sulfate, filtered, and concentrated). The crude product was purified by flash column chromatography (eluting with 50% ethyl acetate in

[1591] 124

[1592] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1593] petroleum ether) to give the title compound (70 mg, 87 %) as a solid. MS (ESI): 777 m / z [M+H]+; LC purity: 95% (254 nm); retention time: 2.26 minutes (LC-MS Method 2).

[1594] 2- / 77-[Bis( tert-butoxycarbonyl )amino ]-16-chloro-18, 19, 25-trifluoro-6, 10,10-trimethyl- 12,12-dioxo-6-phenyl-21-oxa-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-3-yl ] ethyl tert-butyl carbonate

[1595]

[1596]

[0273] Step E: To a stirred solution of Step D product (65 mg, 0.083 mmol) in tetrahydrofuran (5 mL) were added di-tert-butyl dicarbonate (0.058 mL, 0.25 mmol), N, N-Diisopropylethylamine (0.06 mL, 0.332 mmol) and 4-dimethylaminopyridine (4.1 mg, 0.033 mmol). The mixture was stirred at room temperature for 4 hours, diluted with water (20 mL), and extracted with ethyl acetate (20 mL x 2). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (10 g silica gel column; eluting with ethyl acetate in petroleum ether from 0 to 50%) to give the title compound (56 mg, 69 %) as a yellow solid. MS (ESI): 977 m / z [M+H]+; LC purity: 83% (214 nm); retention time: 3.38 minutes (LC-MS Method 4).

[1597] Tert-butyl-N-[ 16-chloro-l 8,19,25-trifluoro-3-( 2-hydroxyethyl)-6, 10, 10-trimethyl-12,12-dioxo-6-phenyl-21-oxa-12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-17-yl Jcarbamate

[1598]

[1599]

[0274] Step F: To a stirred solution of Step E product (56 mg, 0.057 mmol) in methanol (5 mL) was added potassium carbonate (16 mg, 0.12 mmol). The reaction was stirred at room temperature overnight and concentrated. The residue was dissolved in ethyl acetate (20 mL). The solution was washed with brine, dried over anhydrous sodium sulphate, filtered, and

[1600] 125

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[1602] concentrated to give the crude title compound (45 mg, 100 %) as a solid. MS (ESI): 777 m / z [M+H]+; LC purity: 79% (254 nm); retention time: 2.68 minutes (LC-MS Method 4).

[1603] Tert-butyl N-[16-chloro-18,19,25-trifluoro-6,l(),l()-trimethyl-3-[2-(methylamino)ethyl]- 12,12-dioxo-6-phenyl-21-oxa-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl ] carbamate

[1604] BocHN

[1605]

[1606]

[0275] Step G: To a stirred solution of Step F product in dimethyl sulfoxide (5 mL) was added l,l,l-tris(acetyloxy)-l,l-dihydro-l,2-benzodioxol-3-(lH)-one (44 mg, 0.10 mmol). The mixture was stirred at room temperature for 4 hours, diluted with ethyl acetate (40 mL), washed with sodium bicarbonate, aqueous sodium sulfite, brine, dried over sodium sulfate, filtered, and concentrated to give the crude product tert-butyl N-[16-chloro-18, 19, 25-trifluoro-6, 10,10-trimethyl- 12, 12-dioxo-3-(2-oxoethyl)-6-phenyl-21 -oxa- 12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl]carbamate (45 mg) as a yellow solid. MS (ESI): 797 m / z [M+Na]+; LC purity: 35% (254 nm); retention time: 2.55 minutes (LC-MS Method 4).

[1607]

[0276] To a solution of the above crude tert-butylN-[16-chloro-18, 19, 25-trifluoro-6, 10,10-trimethyl- 12,12-dioxo-3-(2-oxoethyl)-6-phenyl-21 -oxa- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-yl]carbamate (37 mg, 0.048 mmol) in methanol (2 mL) was added methanamine (0.12 mL, 0.24 mmol) and 2 drops of acetic acid. The reaction was stirred at room temperature for one hour, treated with sodium cyanoborohydride (9.0 mg, 0.14 mmol), and stirred at room temperature for another 2 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (25 mL). The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica (10 g; eluting with methanol in dichloromethane from 0 to 50%) to give the title compound (11 mg, 29 %) as a solid. MS (ESI): 790 m / z [M+H]+; LC purity: 72% (214 nm); retention time: 2.05 minutes (LC-MS Method 4).

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[1610] Example 17: 16-Chloro-18,19,25-trifluoro-6,10,10-trimethyl-3-[2-(methylamino)ethyl]- 12,12-dioxo-6-phenyl-21-oxa-12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-17-amine

[1611]

[1612]

[0277] Step H: To a stirred solution of Step G product (11 mg, 0.014 mmol) in dichloromethane (2 mL) was added HC1 (4M in dioxane) (2 mL). The mixture was stirred at room temperature for one hour and diluted with ethyl acetate (20 mL). The solution was washed with aqueous sodium bicarbonate, water, brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (1.2 mg, 12 %) as a white solid. MS (ESI): 690 m / z [M+H]+; LC purity: 97% (214 nm); retention time: 2.23 minutes (LC-MS Method 2).1H NMR (500 MHz, CD3OD) 57.43-7.38 (m, 1H), 7.33 (t, J = 9.5 Hz, 1H), 7.29 -7.24 (m, 3H), 7.22 -7.17 (m, 2H), 7.16-7.12 (m, 1H), 4.25 (t, J= 6.0 Hz, 2H), 3.32 (s, 3H), 3.25-3.21 (m, 2H), 3.13-3.08 (m, 2H), 3.02-2.97 (m. 2H), 2.34 -2.28 (m, 2H), 2.24-2.16 (m, 1H), 1.93-1.84 (m, 1H), 1.72 (s, 3H), 1.67-1.558 (m, 1H), 1.49-1.33 (m, 3H), 1.13 (d, 7= 6.5 Hz, 6H) ppm.

[1613] Example 18: (2S)-3-[3-[(6R)-17-amino-16-chloro-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12L6-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1614]

[1615]

[0278] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11) with methyl 5-(4-(bis(tert-butoxycarbonyl)amino)-6-bromo-2,3-difluorophenoxy)-2-fluorobenzimidothioate hydroiodide (3.51 g, 4.88 mmol) and methyl (R)-

[1616] 127

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[1618] 3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (0.95 mL, 4.44 mmol), the reaction condition procedure sequence (Steps A, B, C, D, E described for Example 2, 3 and 4, followed by Step H and I described for Example 1) was followed to prepare the title compound (216 mg) as a white solid. MS (ESI): 733 m / z! M+H]+; LC purity: >99% (214 nm); retention time: 2.03 minutes (LC-MS Method 5).JH NMR (500 MHz, CD3OD) 57.66 (dd, J= 6.5, 2.5 Hz, 1H), 7.52 (dd, 7 = 7.5, 6.0 Hz, 1H), 7.29-7.24 (m, 1H), 7.17 (t, 7= 7.5 Hz, 1H), 7.08 (s, 1H), 7.02 (dd, 7= 10.0, 9.0 Hz, 2H), 4.09-3.95 (m, 2H), 3.83 (d, 7= 2.0 Hz, 3H), 3.16-3.00 (m, 6H), 2.96 (dd, 7= 12.4, 5.6 Hz, 1H), 2.65-2.57 (m, 2H), 2.22 (td, 7 = 13.0, 4.0 Hz, 1H), 1.85 (td, 7 = 13.0, 4.0 Hz, 1H), 1.70 (s. 3H), 1.69-1.60 (m, 1H), 1.20-1.10 (m, 7H), 1.07 (d, 7 = 6.5 Hz, 3H) ppm.

[1619] Example 19: (2S)-3-[3-[(6R)-17-amino-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,16,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1620]

[1621]

[0279] Exchanging 2-bromo-3-fluoro-6-nitropyridine (Intermediate 4-1) with 2-Bromo-3-fluoro-6-nitropyridine (Intermediate 4, 926 mg, 4.19 mmol), the reaction procedure sequences (Steps A, B, C, D, E, F, G and I described for Example 1) was used to prepare the title compound (56 mg, 0.085 mmol, white solid). MS (ESI): 664 m / z [M+H]+; LC purity: 98% (214 nm); retention time: 1.67 minutes (LC-MS Method 2).1H NMR (400 MHz, CD3OD): 57.57-7.51 (m, 2H), 7.44 (t, 7= 9.2 Hz, 1H), 7.34 (dd, 7 = 5.2, 2.8 Hz, 1H), 7.15 (t, 7 = 7.6 Hz, 1H), 7.03-6.94 (m, 3H), 6.85 (d, 7= 9.6 Hz, 1H), 3.86 (d, 7= 2.8 Hz, 3H), 3.48-3.32 (m, 4H), 3.09 (d, J = 14.0 Hz, 1H), 2.99-2.91 (m, 2H), 2.64-2.55 (m, 2H), 2.25 (td, J = 12.4, 4.4 Hz, 1H), 1.84 (td, 7= 12.4, 2.8 Hz, 1H), 1.66 (s, 3H), 1.60-1.44 (m, 3H), 1.14 (s, 3H), 1.07-0.99 (m, 7H) ppm.

[1622] 128

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[1624] Example 20: (2S)-3-[3-[(6R)-17-amino-18-chloro-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12Z6-thia-3,4,16,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1625]

[1626]

[0280] Exchanging 2-bromo-3-fluoro-6-nitropyridine (Intermediate 4-1) with 2-Bromo-3-fluoro-6-nitropyridine (Intermediate 4, 926 mg, 4.19 mmol), the reaction procedure sequences (Steps A, B, C, D, E, F, G, H, and I described for Example 1) was used to prepare the title compound (9.9 mg, 0.014 mmol, white solid). MS (ESI): 698 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.90 minutes (LC-MS Method 5).!H NMR (500 MHz, CD3OD): 5 7.44-7.41 (m, 2H), 7.36 (t, J= 9.0 Hz, 1H), 7.18-7.15 (m, 2H), 7.03-6.94 (m, 3H), 3.82 (d, 7 = 2.0 Hz, 3H), 3.60-3.54 (m, 1H), 3.44-3.38 (m, 1H), 3.14-3.08 (m, 2H), 2.99-2.91 (m, 3H), 2.65-2.57 (m, 2H), 2.22 (td, 7= 12.5, 3.5 Hz, 1H), 1.91 (td, 7= 12.5, 4.0 Hz, 1H), 1.67 (s, 3H), 1.54-1.32 (m, 3H), 1.13 (s, 3H), 1.10-1.05 (m, 7H) ppm.

[1627] Example 21: 6-(3,5-Difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-20-oxa-12k6,18-dithia-3,4,16,26-tetrazatetracyclo[19.3.1.12,5.015,19]hexacosa-l(25),2(26),4,15(19),16,21,23-heptaen-17-amine

[1628]

[1629] Tert-butyl ( tert-butoxycarbonyl)( 4-( 2-( ( 6-( 3,5-difluorophenyl)-6-( 5-( 2-fluoro-5- (methoxy methoxy )phenyl)-l -methyl- 1H-1, 2< 4-triazol-3-yl)-2, 2-dimethylheptyl)sulfonyl)ethyl)thiazol-2-yl)carbamate

[1630]

[1631] 129

[1632] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1633]

[0281] Step A: Exchanging methyl 5-((2-(bis(tert-butoxycarbonyl)amino)-4-bromothiazol-5-yl)thio)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-2) with methyl 2-fluoro-5-(methoxymethoxy)benzimidothioate hydroiodide (Intermediate 5, 0.61 g, 1.71 mmol), the reaction procedure sequence (Steps A, B, C (tert-butyl (4-bromothiazol-2-yl)(tert-butoxycarbonyl)carbamate (Intermediate 6 as bromide coupling partner) described for Example 10, Step F described for Example 1) was used to prepare the title compound (900 mg, 1.04 mmol ) as a yellow solid. MS (ESI): 866 m / z [M+H]+; LC purity: 85% (214 nm); retention time: 2.31 minutes (LC-MS Method 2).

[1634] Tert-butyl ( 5-bromo-4-( 2-((6-(3, 5 -difluorophenyl )-6-( 5-( 2-fluoro-5- (methoxymethoxy )phenyl)-l -methyl- 1H-1, 2,’ 4-triazol-3-yl)-2, 2-dimethylheptyl)sulfonyl)ethyl)thiazol-2-yl)(tert-butoxycarbonyl)carbamate

[1635]

[1636]

[0282] Step B: To a stirred solution of Step A product (170 mg, 0.196 mmol) in acetonitrile (3 mL) was added N-bromosuccinimide (70 mg, 0.393 mmol). The reaction was stirred at room temperature for 2 days, quenched with 30 mL of water, and extracted with ethyl acetate (30 mL x 3). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0-70% of ethyl acetate in petroleum ether) to give the title compound (120 mg, 0.127 mmol, 65 %) as a yellow solid. MS (ESI): 944, 946 m / z [M+H]+; LC purity: 90% (214 nm); retention time: 2.43 minutes (LC-MS Method 2).

[1637] 3-(3-(7-((2-(2-Amino-5-bromothiazol-4-yl)ethyl)sulfonyl)-2-(3,5-difluorophenyl)-6,6-dimethylheptan-2-yl)-l-methyl-lH-l,2,4-triazol-5-yl)-4-fluorophenol

[1638]

[1639]

[0283] Step C: To a stirred solution of Step B product (120 mg, 0.127 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2.5 mL). The reaction mixture was

[1640] 130

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[1642] stirred at room temperature for 2 hours and concentrated. The residue was basified to pH ~ 8 with sodium bicarbonate and extracted with ethyl acetate (30 mL x 2). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated to give the crude title compound (80 mg, 0.114 mmol, 89 %) as a yellow solid. MS (ESI): 700, 702 m / z [M+H]+; LC purity: 72% (214 nm); retention time: 1.97 minutes (LC-MS Method 2).

[1643] 6-(3,5-Difluorophenyl)-24-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-20-oxa-12X6,18-dithia-3, 4,16, 26-tetrazatetracyclo[19.3.1.12, 5.015, 19]hexacosa-l(25), 2(26), 4, 15(19), 16, 21,23-heptaen-17-amine

[1644]

[1645]

[0284] Step D: To a stirred solution of Step C product (100 mg, 0.143 mmol) in acetone (3 mL) was added potassium carbonate (59 mg, 0.428 mmol). The mixture was stirred at 70 °C for 3 hours and concentrated. The residue was diluted with ethyl acetate (30 mL), washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (5.2 mg, 0.0084 mmol, 5.9 %) as a white solid. MS (ESI): 620 m / z [M+H]+; LC purity: 95% (214 nm); retention time: 1.92 minutes (LC-MS Method 2). 'H NMR (400 MHz, CD3OD) 8 7.55-7.47 (m, 1H), 7.42-7.30 (m, 2H), 6.92-6.81 (m, 2H), 6.78-6.68 (m, 1H), 3.82 (s, 3H), 3.45-3.36 (m, 1H), 3.26-3.18 (m, 1H), 3.02-2.84 (m, 4H), 2.35-2.19 (m, 1H), 2.02-1.86 (m, 1H), 1.70 (s, 3H), 1.62-1.45 (m, 2H), 1.39-1.26 (m, 2H), 1.16 (s, 3H), 1.05 (s, 3H) ppm.

[1646] Example 22: (2S)-3-[3-[(6R)-17-amino-16, 18,19, 25-tetrafluoro-3, 6,10, 10-tetramethyl-12,12-dioxo-21-oxa-12i6-thia-3,4,27-triazatetracyclo [20.3.1.12,5.015,20] heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl] phenyl]-2-methyl-propanoic acid

[1647]

[1648] 131

[1649] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1650]

[0285] Exchanging 4-bromo-5-fluoro-2-nitro-pyridine (Intermediate 4-1) with 1-bromo-2,3,4,6-tetrafluoro-5-nitrobenzene (9.50 g, 34.7 mmol), the reaction procedure sequence, Steps A, B, C, D, E, F, G, and I, described in Example 1, was used to prepare the title compound (1.68 g, 2.35 mmol) as a white solid. MS (ESI): 717 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.97 minutes (LC-MS Method 5).JH NMR (400 MHz, DMSO-rfe) 5 12.12 (s, 1H), 7.43 (t, J = 9.6 Hz, 1H), 7.39-7.37 (m, 1H), 7.32-7.29 (m, 1H), 7.14 (t, J = 7.6 Hz, 1H), 7.07 (s, 1H). 6.98-6.95 (m, 2H), 5.61 (s, 2H), 3.77 (s, 3H), 3.33-3.22 (m, 2H), 3.06-3.00 (m, 2H), 2.95-2.87 (m, 2H), 2.86-2.79 (m, 2H), 2.58-2.52 (m, 1H), 2.16-2.11 (m, 1H), 1.74-1.68 (m, 1H), 1.63 (s, 3H), 1.58-1.48 (m, 1H), 1.32-1.23 (m, 2H), 1.08-0.92 (m, 10H) ppm.

[1651] Example 23: Methyl (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1652]

[1653]

[0286] Exchanging 4-bromo-5-fluoro-2-nitro-pyridine (Intermediate 4-1) with 4-bromo-2,3,5-trifluoro-6-nitropyridine (360 mg, 1.40 mmol), the reaction procedure sequence, Steps A, C, B, D, E, G and F described in Example 1 (in this order), was used to prepare the title compound (1.5 mg, 0.00210 mmol) as a dark yellow solid. MS (ESI): 714 m / z [M+H]+; LC purity: 97% (214 nm); retention time: 2.52 minutes (LC-MS Method 4).

[1654] Example 24 and 25: Diastereomers 1 and 2 of (2S)-3-[3-[(6R)-17-amino-16,18,19,25-tetrafluoro-21-hydroxy-3, 6,10, 10-tetramethyl-12,12-dioxo-12X6-thia-3, 4,27-

[1655] 132

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[1657] triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1658]

[1659] Example 26: (2S)-3-[3-[(6R)-17-amino-16, 18,19, 25-tetrafluoro-3, 6,10, 10-tetramethyl- 12,12,21-trioxo-12k6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1660]

[1661] Example 27: (6R)-17-amino-16,18,19,25-tetrafluoro-6-[3-[(2S)-3-hydroxy-2-methyl-propyl]phenyl]-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-21-one

[1662]

[1663]

[0287] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1) (4-(bis(tert-butoxycarbonyl)amino)-2-bromo-3,5,6-trifluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (1.2 g, 1.52 mmol), the reaction procedure sequence, Steps A, B, C, D, E described for Examples 5, 6, 7 and 8, followed by Steps H described for Example 1, and Steps F and G described for Examples 5, 6, 7 and 8, was used to prepare the title compounds.

[1664]

[0288] Example 24: Eluent 1, Diastereomer 1 of (2S)-3-[3-[(6R)-17-amino-16, 18, 19,25-tetrafluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6- 133

[1665] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1666] yljphenyl] -2 -methyl-propanoic acid (19 mg, white solid). MS (ESI): 731 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.93 minutes (LC-MS Method 2).1H NMR (500 MHz, CD3OD) 57.89 (S, 1H), 7.42 (S, 1H), 7.27 (t, J = 9.0 Hz, 1H), 7.18 (t, 7 = 7.5 Hz, 1H), 7.08 (s, 1H), 7.01 (d, 7 = 8.0 Hz, 2H), 6.32 (s, 1H), 3.85 (s, 3H), 3.12-3.03 (m, 2H), 3.01 (s, 2H), 2.96 (dd, 7 = 12.5, 5.5 Hz, 1H), 2.86 (s, 1H), 2.67-2.55 (m, 2H), 2.36-2.27 (m, 1H), 1.89-1.80 (m, 1H), 1.71 (s, 4H), 1.50-1.35 (m, 2H), 1.34-1.26 (m, 1H), 1.18 (s, 1H), 1.12 (s, 6H), 1.07 (d, J = 6.5 Hz, 3H) ppm.

[1667]

[0289] Example 25: Eluent 2, Diastereomer 2 of (2S)-3-[3-[(6R)-17-amino-16, 18, 19,25-tetrafluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (25 mg, white solid). MS (ESI): 731 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.95 minutes (LC-MS Method 2). ’H NMR (500 MHz, CD3OD) 5 8.00 (d, 7 = 6.5 Hz, 1H), 7.26 (d, 7 = 8.5 Hz, 2H), 7.13 (dd, 7 = 8.5, 6.5 Hz, 2H), 7.01 (dd, 7= 18.5, 7.5 Hz, 2H), 6.25 (s, 1H), 3.89 (d, 7= 3.0 Hz, 3H), 3.47-3.39 (m, 1H), 3.06 (d, 7 = 14.0 Hz, 1H), 2.99-2.92 (m, 3H), 2.88-2.79 (m, 2H), 2.65-2.58 (m, 2H), 2.37-2.28 (m, 1H), 2.17-2.07 (m, 1H), 1.71 (s, 3H), 1.54-1.47 (m, 1H), 1.46-1.36 (m, 2H), 1.29 (s, 3H), 1.15 (s, 3H), 1.08 (d, 7= 6.5 Hz, 3H), 1.05-0.98 (m, 1H) ppm.

[1668]

[0290] Example 26: (2S)-3-[3-[(6R)-17-amino-16,18,19,25-tetrafluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (25 mg, white solid). MS (ESI): 729 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.98 minutes (LC-MS Method 4). ‘H NMR (500 MHz, CD3OD) 5 8.17-8.12 (m, 1H), 7.97 (dd, 7= 6.5, 2.0 Hz, 1H), 7.55 (t, J = 9.0 Hz, 1H). 7.16 (t, 7 = 7.5 Hz, 1H), 7.04 (s, 1H), 7.00 (t, 7 = 7.5 Hz, 2H), 3.86 (d, 7 = 2.0 Hz, 3H), 3.24-3.16 (m, 3H), 3.14-3.08 (m, 2H), 2.95-2.89 (m, 1H), 2.73 (d, 7 = 14.0 Hz, 1H), 2.63-2.56 (m, 2H), 2.24-2.16 (m, 1H), 1.91-1.84 (m, 1H), 1.68 (s, 3H), 1.64-1.52 (m, 1H), 1.46-1.36 (m, 1H), 1.30-1.23 (m, 1H), 1.07-1.02 (m, 9H), 1.01-0.94 (m, 1H) ppm.

[1669]

[0291] Example 27: (6R)-17-amino-16,18,19,25-tetrafluoro-6-[3-[(2S)-3-hydroxy-2-methyl-propyllpheny 1] - 3, 6, 10, 10-tetramethyl- 12,12-dioxo- 12Z6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-21-one (2.1 mg, white solid). MS (ESI): 715 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.09 minutes (LC-MS Method 4). ’H NMR (500 MHz, CD3OD) 58.09-8.01 (m, 1H), 7.85 (dd, 7 = 6.5, 2.0 Hz, 1H), 7.46 (t, 7 = 9.0 Hz, 1H), 7.06 (t, 7 = 7.5 Hz, 1H), 6.93-6.85 (m, 3H), 3.77 (d, 7 = 2.0 Hz, 3H), 3.31-3.23 (m, 2H), 3.16-3.12 (m, 2H), 3.11-3.05 (m, 1H), 3.04-2.98 (m,

[1670] 134

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[1672] 1H), 2.98-2.94 (m, 1H), 2.67-2.59 (m, 2H), 2.18 (dd, J = 13.5, 8.5 Hz, 1H), 2.11 (t, J = 11.0 Hz, 1H), 1.80 (td, J = 13.0, 5.0 Hz, 1H), 1.73-1.65 (m, 1H), 1.59 (s, 3H), 1.56-1.45 (m, 1H), 1.38-1.28 (m, 1H), 1.27-1.13 (m, 1H), 0.95 (d, J = 3.0 Hz, 6H), 0.93-0.85 (m, 1H), 0.69 (d, J = 6.5 Hz, 3H) ppm.

[1673] Example 28 and 29: Diastereomers 1 and 2 of (2S)-3-[3-[(6R)-18-amino-16,17,19,25-tetrafluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1674]

[1675] Example 30: (2S)-3-[3-[(6R)-18-amino-16, 17,19, 25-tetrafluoro-3, 6,10, 10-tetramethyl- 12,12, 21-trioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1676]

[1677]

[0292] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1) with methyl 5-((3-(bis(tert-butoxycarbonyl)amino)-6-bromo-2,4,5-trifluorophenyl)(hydroxy)methyl)-2-fluorobenzimidothioate hydroiodide (1.05 g, 1.40 mmol), the reaction procedure sequence, Steps A, B described for Examples 5, 6, 7 and 8, followed by reaction conditions described for Intermediate 11C, then Steps C as described for Example 5, 6, 7, 8 and Step H described for Example 1, and lastly Steps E, F and G described for Examples 5, 6, 7 and 8, was used to prepare the title compounds.

[1678]

[0293] Example 28: Eluent 1, Diastereomer 1 of (2S)-3-[3-[(6R)-18-amino-16, 17, 19,25-tetrafluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo-l2Z6-thia-3,4.27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-

[1679] 135

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[1681] yljphenyl] -2 -methyl-propanoic acid (8.0 mg, white solid). MS (ESI): 731 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.84 minutes (LC-MS Method 5).1H NMR (500 MHz, CD3OD) 58.00 (d, J= 6.5 Hz, 1H), 7.28 (d, J = 8.0 Hz, 2H), 7.15-7.12 (m, 2H), 7.02 (dd, J = 20.5, 7.5 Hz, 2H), 6.29 (s, 1H), 3.89 (d, 7= 3.0 Hz, 3H), 3.47-3.43 (m, 1H), 3.06 (d, 7= 14.0 Hz, 1H), 2.99-2.94 (m, 3H), 2.86 (dd, 7= 19.0, 8.0 Hz, 2H), 2.63-2.57 (m, 2H), 2.37-2.29 (m, 1H), 2.16-2.06 (m, 1H), 1.71 (s, 3H), 1.58-1.32 (m, 4H), 1.29 (s, 3H), 1.16 (s, 3H), 1.08 (d, 7 = 6.5 Hz, 3H) ppm.

[1682]

[0294] Example 29: Eluent 2, Diastereomer 2 of (2S)-3-[3-[(6R)-18-amino-16, 17, 19,25-tetrafluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12X6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (8.4 mg, white solid). MS (ESI): 731 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.86 minutes (LC-MS Method 5). ’H NMR (500 MHz, CD3OD) 58.00 (d, 7= 6.0 Hz, 1H), 7.30-7.26 (m, 2H), 7.14 (dd, 7 = 4.0, 3.0 Hz, 2H), 7.02 (dd, J = 20.5, 7.5 Hz, 2H), 6.29 (s, 1H), 3.89 (d, 7 = 3.0 Hz, 3H), 3.47-3.42 (m, 1H), 3.06 (d, 7 = 14.0 Hz, 1H), 3.00-2.94 (m, 3H), 2.86 (dd, 7= 18.5, 8.0 Hz, 2H), 2.63-2.57 (m, 2H), 2.37-2.29 (m, 1H), 2.14-2.07 (m, 1H). 1.71 (s, 3H), 1.53-1.38 (m, 3H), 1.29 (s, 3H). 1.16 (s, 3H), 1.09-1.02 (m, 4H) ppm.

[1683]

[0295] Example 30: (2S)-3-[3-[(6R)-18-amino-16,17,19,25-tetrafluoro-3,6,10,10-tetramethyl-12.12, 21 -trioxo- 12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (6.2 mg, white solid). MS (ESI): 729 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.92 minutes (LC-MS Method 5). ‘H NMR (500 MHz, CD3OD) 5 8.17-8.12 (m, 1H), 7.97 (dd, 7= 6.5, 2.0 Hz, 1H), 7.55 (t, 7= 9.0 Hz, 1H), 7.16 (t, 7= 7.2 Hz, 1H), 7.05-6.98 (m, 3H), 3.86 (d, 7= 2.0 Hz, 3H), 3.20 (ddd, 7 = 17.0, 9.5, 4.0 Hz, 3H), 3.15-3.05 (m, 2H), 2.99-2.89 (m, 1H), 2.73 (d, 7= 14.0 Hz, 1H), 2.62-2.54 (m, 2H), 2.23-2.17 (m, 1H), 1.87 (tt, 7 = 7.0, 3.5 Hz, 1H), 1.67 (s, 3H), 1.63-1.57 (m, 1H), 1.43-1.25 (m, 3H), 1.06-1.03 (m, 9H) ppm.

[1684] Example 31: (2S)-3-[3-[(6R)-18-amino-16,17,19,25-tetrafluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1685] 136

[1686] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1687]

[1688]

[0296] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1) with methyl 5-(3-amino-6-bromo-2,4,5-trifluorobenzyl)-2-fluorobenzimidothioate hydroiodide (0.68 g, 1.27 mmol), the reaction procedure sequence, Steps A, B described for Examples 5, 6, 7 and 8, followed by reaction conditions described for Intermediate 11 A, then Steps C, D, E and F as described for Example 5, 6, 7, 8, was used to prepare the title compound. MS (ESI): 715 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.93 minutes (LC-MS Method 5).

[1689]

[1690] NMR (500 MHz, CD3OD) 57.66 (dd, 7 = 6.5, 2.5 Hz, 1H), 7.52 (dd, J = 7.5, 6.0 Hz, 1H), 7.29-7.24 (m, 1H), 7.17 (t, J = 7.5 Hz, 1H), 7.08 (s, 1H), 7.02 (dd, J= 10.0, 9.0 Hz, 2H), 4.09-3.95 (m, 2H), 3.83 (d, J = 2.0 Hz, 3H), 3.16-3.00 (m, 6H), 2.96 (dd, J = 12.4, 5.6 Hz, 1H), 2.65-2.57 (m, 2H), 2.33 (td, J = 13.0, 4.0 Hz, 1H), 1.91 (td, J = 13.0, 4.0 Hz, 1H), 1.70 (s, 3H), 1.64-1.48 (m, 3H), 1.17 (d, J = 15.5 Hz, 7H), 1.07 (d, J= 6.5 Hz, 3H) ppm.

[1691] Example 32: (2S)-3-[3-[(6R)-16-amino-19,25-difluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,17,27-tetrazatetracyclo[20.3. L12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1692]

[1693]

[0297] Exchanging 4-bromo-5-fluoro-2-nitro-pyridine (Intermediate 4-1) with tert-butyl (tert-butoxycarbonyl)(4,5-difluoro-3-iodopyridin-2-yl)carbamate (271 mg, 0.594 mmol), the reaction procedure sequence, Steps A, B, E, F, G, and I described for Examples 1, was used to prepare the title compound (4 mg, white solid). MS (ESI): 682 m / z [M+H]+; LC purity: 95% (214 nm); retention time: 2.05 minutes (LC-MS Method 4). 'H NMR (400 MHz, CD3OD) 5 7.83 (d, J = 2.8 Hz, 1H), 7.51-7.47 (m, 1H), 7.40 (t, J = 8.8 Hz, 1H), 7.23 (dd, J = 3.2 Hz, 1H), 7.17-7.13 (m, 1H), 7.02-6.98 (m, 2H), 6.93 (d, J = 8.0 Hz, 1H), 3.84 (d, J = 2.0 Hz, 3H), 3.17- 137

[1694] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1695] 3.06 (m, 4H), 2.97-2.92 (m, 1H), 2.88-2.85 (m, 1H), 2.63-2.54 (m, 2H), 2.23-2.17 (m, 1H), 1.85-1.77 (m, 1H), 1.72-1.67 (m, 4H), 1.55-1.45 (m, 1H), 1.37-1.30 (m, 2H), 1.13-1.04 (m, 10H) ppm.

[1696] Example 33: Methyl (2S)-3-[3-[(6R)-17-amino-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12Z6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1697] F I

[1698] N>

[1699]

[1700]

[0298] The crude product from Step G of Example 1 was further purified by prep-HPLC to afford the purified title compound. (70 mg, 0.103 mmol) as a white solid. MS (ESI): 678 m / z [M+H]+; LC purity: 90% (214 nm); retention time: 1.66 minutes (LC-MS Method 2).JH NMR (400 MHz, CD3OD) 57.50 (s, 1H), 7.48-7.35 (m, 3H), 7.16 (t, J= 7.6 Hz, 1H), 7.05-6.93 (m, 3H), 6.82 (s, 1H), 3.86 (s, 3H), 3.56 (s, 3H), 3.42-3.36 (m, 2H), 3.17-3.12 (m, 2H), 3.09-2.86 (m. 3H), 2.71-2.59 (m, 2H), 2.29-2.18 (m, 1H), 1.90-1.75 (m, 1H), 1.67 (s, 3H), 1.64-1.52 (m, 1H), 1.46-1.30 (m, 3H), 1.14-1.04 (m, 9H) ppm.

[1701] Example 34: Methyl (2S)-3-[3-[(6R)-16-amino-17, 18,19, 25-tetrafluoro-3, 6, 10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1702]

[1703]

[0299] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fhioro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11) with methyl 5-(3-amino-2-bromo-4,5,6-trifluorophenoxy)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-11, 400 mg, 0.745 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-

[1704] 138

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[1706] oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl- 1 -(2-methylhydrazineyl)- 1 -oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9-2, 361 mg, 0.745 mmol), the reaction procedure sequence, Step A, B described for Example 2, followed by reaction conditions described for Intermediate 11 A, then Steps C, D, E described for Example 2, was used to prepare the title compound (1.3 mg, white solid). MS (ESI): 731 nz / z [M+H]+; LC purity: 93% (214 nm); retention time: 2.73 minutes (LC-MS Method 4).1H NMR (400 MHz, CD3OD) d 7.44-7.36 (m, 2H), 7.22 (dd, J = 5.2, 2.8 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.01-6.95 (m, 3H), 3.84 (d, J = 2.0 Hz, 3H), 3.54 (s, 3H), 3.28-3.09 (m, 5H), 2.89-2.85 (s, 2H), 2.69-2.60 (m, 2H), 2.21-2.15 (m, 1H), 1.81 (td, 7 = 13.2, 4.4 Hz, 1H), 1.67 (s, 3H), 1.70-1.60 (m, 1H), 1.52-1.45 (m, 1H), 1.34-1.31 (m, 1H), 1.11-1.01 (m, 1 OH) ppm.

[1707] Example 35: Diastereomer 1 of (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1708] Example 36: Diastereomer 2 of (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-21-hydroxy-3,6,10,10-tetramethyl-12,12-dioxo-12i6-thia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1709]

[1710] Example 37: (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12X6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1711]

[1712] 139

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[1714]

[0300] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11) with (4-(bis(tert-butoxycarbonyl)amino)-2-bromo-3,6-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-12, 1.34 g, 1.73 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9-2, 0.70 g, 1.44 mmol), the reaction procedure sequence (Step A, B, C, D, E, F, G, and H described for Example 2, 3 and 4) was used to prepare the title compounds.

[1715]

[0301] Example 35: Eluent 1, Diastereomer 1 of (2S)-3-[3-[(6R)-17-amino-16, 19,25-trifluoro-21 -hydroxy-3,6, 10,10-tetramethyl- 12,12-dioxo- 12Z6-thia-3,4,27 -triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl] -2 -methyl-propanoic acid (10 mg, white solid). MS (ESI): 713 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.91 minutes (LC-MS Method 2).1H NMR (400 MHz, CD3OD) 5 7.97-7.88 (m, 1H), 7.45-7.38 (m, 1H), 7.28-7.24 (m, 1H), 7.18 (t, J= 7.6 Hz, 1H).

[1716] 7.10-7.07 (m, 1H), 7.03-6.99 (m, 2H), 6.52-6.47 (m, 1H), 6.41 (s, 1H), 3.85 (s, 3H), 3.13-2.79 (m, 7H), 2.67-2.57 (m, 2H), 2.35-2.26 (m, 1H), 1.87-1.80 (m, 1H), 1.76-1.66 (m, 1H), 1.71 (s, 3H), 1.50-1.28 (m, 3H), 1.19 (s, 3H), 1.11 (s, 3H), 1.08 (d, 7= 6.4 Hz, 3H) ppm.

[1717]

[0302] Example 36: Eluent 2, Diastereomer 2 of (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-21 -hydroxy-3,6, 10, 10-tetramethyl- 12, 12-dioxo- 12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl] -2 -methyl-propanoic acid (12 mg, white solid). MS (ESI): 713 m z [M+H]+; LC purity: >99% (214 nm); retention time: 1.94 minutes (LC-MS Method 2).1H NMR (400 MHz, CD3OD) 5 8.02-7.98 (m, 1H), 7.26-7.22 (m, 2H), 7.13-7.06 (m, 2H), 7.02-6.95 (m, 2H), 6.50-6.44 (m, 1H), 6.35-6.25 (m, 1H), 3.88 (d, J = 2.8 Hz, 3H), 3.05 (d, J = 14.0 Hz, 1H), 2.93-2.72 (m, 6H), 2.65-2.57 (m, 2H), 2.36-2.29 (m, 1H), 2.18-2.08 (m, 1H), 1.70 (s, 3H), 1.54-1.36 (m, 3H), 1.30 (s, 3H), 1.15 (s, 3H), 1.09-1.07 (m, 3H), 1.01-0.96 (m, 1H) ppm.

[1718]

[0303] Example 37: (2S)-3-[3-[(6R)-17-amino-16,19,25-trifluoro-3,6,10,10-tetramethyl- 12,12,21-trioxo-12 6-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid (2.0 mg, white solid). MS (ESI): 711 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.30 minutes (LC-MS Method 4). ’H NMR (400 MHz, CD3OD) 58.28-8.24 (m, 1H), 8.00 (dd, J= 2.0, 6.4 Hz, 1H), 7.57 (t, J = 9.2 Hz, 1H), 7.16 (t, J = 8.0 Hz, 1H), 7.04-6.99 (m, 3H), 6.67 (dd, J = 7.6,

[1719] 140

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[1721] 11.6 Hz, 1H), 3.84 (d, 7 = 2.0 Hz. 3H), 3.22-3.15 (m, 2H), 3.11-3.04 (m, 2H), 2.99-2.90 (m, 3H), 2.80 (d, J= 14.0 Hz, 1H), 2.64-2.56 (m, 2H), 2.23-2.18 (m, 1H), 1.92-1.85 (m, 1H), 1.68 (s, 3H), 1.61-1.52 (m, 1H), 1.41-1.28 (m, 2H), 1.10-1.04 (m, 9H) ppm.

[1722] Example 38: (2S)-3-[3-[(6R)-17-Amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1723] F

[1724] , /

[1725] U

[1726]

[1727] Methyl ( 2S)-3-[ 3-[ ( 6R)-17-[his( tert-hutoxycarhonyl)amino]-18, 19,25-trifluoro-3,6,10, 10-tetramethyl-12,12-dioxo-12 6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl ]phenyl ]-2-methyl-propanoate

[1728]

[1729]

[0304] Step A: Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-4,5-dichloro-2-fluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11) with methyl 5-((4-amino-6-bromo-2,3-difluorophenyl)thio)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-13, 7 g, 14.3 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9-2, 6.3 g, 12.99 mmol), the reaction procedure sequence, Step A, B, C, D, and E described for Example 9, was used to prepare the title compound (550 mg, 0.48 mmol) as a white solid. MS (ESI): 929 m / z [M+H]+

[1730] 141

[1731] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1732] Methyl (2S)-3-[3-[(6R)-17-ammo-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo- 12X6,2 l-dithia-3, 4, 27-triazatetracyclo[ 20.3.1.12,5.015,20 ]heptacosa-1(26),2(27),4,15,17,19,22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoate

[1733]

[1734]

[0305] Step B: The title compound (350 mg, 0.48 mmol) was prepared as a white solid from Step A product (550 mg, 0.48 mmol) following the reaction conditions described in Step G of Example 9. MS (ESI): 729 m / z [M+H]+; LC purity: 90% (214 nm); retention time: 2.00 minutes (LC-MS Method 3).

[1735] Example 38: (2S)-3-[3-[(6R)-17-Amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1736]

[1737]

[0306] Step C: Utilizing the reaction conditions described in Step I of Example 1, the title compound (29 mg, 59%) was prepared from Step B product (50 mg, 0.069 mmol) as a white solid. MS (ESI): 715 m / z. [M+H]+; LC purity: >99% (214 nm); retention time: 2.02 minutes (LC-MS Method 3). ‘H NMR (500 MHz, CDsOD) 57.76-7.71 (m, 1H), 7.64 (dd, 7= 6.5, 2.5 Hz, 1H), 7.31 (t, 7= 9.0 Hz, 1H), 7.16 (t, 7 = 7.5 Hz, 1H), 7.06 (s, 1H), 7.01 (t, 7 = 7.5 Hz, 2H), 6.60 (d, 7 = 7.0 Hz, 1H), 3.77 (d, 7= 1.5 Hz, 3H), 3.34-3.31 (m, 1H), 3.25-3.18 (m, 3H), 3.00-2.90 (m, 3H), 2.65-2.53 (m, 2H), 2.31-2.23 (m, 1H), 1.95-1.85 (m, 1H). 1.69 (s, 3H), 1.60-1.50 (m, 1H), 1.47-1.39 (m, 2H), 1.15 (d, 7= 13.0 Hz, 6H), 1.14-1.10 (m, 1H), 1.06 (d, 7= 6.5 Hz, 3H) ppm.

[1738] Example 39: (2S)-3-[3-[(6R)-17-Amino-18,19,25-trifluoro-16-iodo-3,6,10,10-tetramethyl- 12,12-dioxo-12k6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1739] 142

[1740] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1741]

[1742] Methyl ( 2S)-3-[ 3-[ ( 6R)-17-amino-18, 19,25-trifluoro-16-iodo-3, 6, 10, 10-tetramethyl-12, 12-dioxo-12 6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26), 2(27), 4, 15,17,19,22,24-octaen-6-yl ] phenyl ]-2-methyl-propanoate

[1743]

[1744]

[0307] Step A: To a stirred solution of methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6, 10, 10-tetramethyl-12, 12-dioxo- 12X6,21 -dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l (26), 2(27), 4, 15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step B product of Example 38, 300 mg, 0.411 mmol) in acetic acid (8 mL) was added N-iodosuccinimide (0.095 g, 0.411 mmol). The reaction mixture was stirred at room temperature for 2 hours and concentrated. The residue was dissolved in ethyl acetate (60 mL), washed with saturated sodium bicarbonate, water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (40 g silica gel column, eluting with 0-60% ethyl acetate in petroleum) to give the title compound (0.18 g, 51%) as a white solid. MS (ESI): 855 m / z [M+HJ+; LC purity: 90% (254 nm); retention time: 2.24 minutes (LC-MS Method 5).

[1745] Example 39: ( 2S)-3-[ 3-[ (6R)-17-Amino-l 8,19, 25-trifluoro-16-iodo-3,6,10, 10-tetramethyl- 12,12-dioxo-1276,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26), 2(27), 4, 15,17,19,22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoic acid

[1746]

[1747] 143

[1748] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1749]

[0308] To a solution of Step A product (50 mg, 0.059 mmol) in tetrahydrofuran (3 mL) was added lithium hydroxide (3.0 mL, 3.0 mmol, 1 M in water). The reaction was stirred at room temperature for 16 hours, acidified with 1.0 M hydrochloric acid to pH ~ 5, and extracted with ethyl acetate (20 x 3 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by prep-HPLC to give the title compound (29.8 mg, 60 %) as a white solid. MS (ESI): 841 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.10 minutes (LC-MS Method 3).1H NMR (500 MHz, CD3OD) 57.94 (dd, 7= 6.5, 2.5 Hz, 1H), 7.81-7.71 (m, 1H), 7.34 (t, 7= 9.5 Hz, 1H), 7.17 (t, 7= 7.5 Hz, 1H), 7.07 (s, 1H), 7.02-6.99 (m, 2H), 3.84 (d, 7= 2.0 Hz, 3H), 3.73-3.62 (m, 2H), 3.19-3.10 (m, 2H), 3.04 (s, 2H), 2.97-2.92 (m, 1H), 2.65-2.56 (m, 2H), 2.39-2.24 (m, 1H), 1.99-1.86 (m, 1H), 1.71 (s, 3H), 1.67-1.58 (m, 1H), 1.56-1.45 (m, 2H), 1.36-1.22 (m, 1H), 1.19 (d, 7 = 23 Hz, 6H), 1.07 (d, 7= 6.5 Hz, 3H) ppm.

[1750] Example 40: (2S)-3-[3-[(6R)-17-Amino-18,19,25-trifluoro-3,6,10,10,16-pentamethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1751] F

[1752] , /

[1753] Methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifhioro-3,6,10,10,16-pentamethyl-12,12-dioxo-12X6,21 -dithia-3, 4, 27 -triazatetracyclol 20.3.1.12,5.015,20 ]heptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoate, F, / 1— u

[1754]

[1755]

[0309] Step A: To a stirred solution of methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-16-iodo-3,6,10,10-tetramethyl-12,12-dioxo-12L6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step A product of Example 39, 120 mg, 0.14 mmol) and methylboronic acid (122 mg, 2.04 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was 144

[1756] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1757] added tetrakis (triphenylphosphine)palladium (15.9 mg, 0.013 mmol) and potassium carbonate (188 mg, 1.36 mmol). The mixture was purged with nitrogen, stirred at 90 °C for 16 hours, and concentrated. The residue was diluted with ethyl acetate, washed with water, brine, dried over anhydrous sodium sulphate, filtered, and concentrated. The crude product was purified by flash chromatography (25g silica gel column, eluting with 0-30% ethyl acetate in petroleum ether) to give the title compound (50 mg, 48 %) as a solid. MS (ESI): 743 m / z [M+H]+; LC purity: 90% (214 nm); retention time: 2.18 minutes (LC-MS Method 2).

[1758] Example 40: ( 2S)-3-[ 3-[ (6R)-17-Amino-l 8, 19, 25-trifluoro-3, 6,10,10,16-pentamethyl-12, 12-dioxo- 12X6, 21 -dithia-3,4, 27-triazatetracyclo[20.3.1.12,5.015, 20]heptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl Jphenyl ]-2-methyl-propanoic acid

[1759] , F

[1760] I

[1761] >'>-N

[1762]

[1763]

[0310] Step B: Utilizing the reaction conditions described in Step I of Example 1, the title compound (40.7 mg, 83%) was prepared from Step A product (50 mg, 0.069 mmol) as a white solid. MS (ESI): 729 mJ [M+H]+; LC purity: >99% (214 nm); retention time: 2.07 minutes (LC-MS Method 3). ‘H NMR (500 MHz, DMSO-de) 57.82 (d, J = 4.5 Hz, 1H), 7.69-7.57 (m, 1H), 7.44 (t, 7 = 9.5 Hz, 1H), 7.14 (t, 7 = 7.5 Hz, 1H), 7.09 (s, 1H), 6.98-6.93 (m, 2H), 5.76 (s, 2H), 3.76 (d, J = 2.0 Hz, 3H), 3.31-3.25 (m, 2H), 3.05-2.84 (m, 5H), 2.60-2.52 (m, 2H), 2.23 (t, J = 10.5 Hz, 1H), 2.04 (s, 3H), 1.75 (t, J = 11.0 Hz, 1H), 1.65 (s, 3H), 1.54-1.30 (m, 3H), 1.14 (s, 3H), 1.12-1.08 (m, 1H), 1.07 (s, 3H), 0.97 (d, 7= 6.5 Hz, 3H) ppm.

[1764] Example 41 and 42: Diastereomers 1 and 2 of (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-1216,21X4-dithia-3,4,27-

[1765] 145

[1766] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1767] triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1768] Diastereomers of methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-1276,2124-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1769]

[1770]

[0311] Step A: To a stirred solution of methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-16-iodo-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step B product of Example 39, 0.34 g, 0.467 mmol) in acetonitrile (5 mL) was added p-toluenesulfonic acid (10 mg, catalytic) and hydrogen peroxide (0.5 mL, 75% in water). The reaction mixture was stirred at room temperature for 7 days, diluted with water (5 mL), and extracted with ethyl acetate (5 mL x 2). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (20 g silica gel column, eluting with 0-60% ethyl acetate in petroleum ether) to give the title compound (170 mg, 50 %) as a yellow solid. MS (ESI): 745 m / z. [M+H]+; LC purity: 95% (214 nm); retention time: 2.00 minutes (LC-MS Method 3).

[1771] Example 41 and 42: Diastereomers 1 and 2 of (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10, 10-tetramethyl-12, 12,21-trioxo-12X6,21X4-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl Jphenyl ]-2-methyl-propanonic acid

[1772] 146

[1773] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1774]

[1775]

[0312] Step B: Utilizing the reaction conditions described in Step I of Example 1, the title compounds were prepared from Step A product (85 mg, 0.114 mmol) as a white solid. The title compound mixture was separated by prep-HPLC. The first eluent, the Diastereomer 1, was designated as Example 41 (13.2 mg, 15 %); The second eluent, the Diastereomer 2, was designated as Example 42 (34.3 mg, 40%).

[1776]

[0313] Example 41: Diastereomer 1, MS (ESI): 731 m / z. [M+H]+; LC purity: >99% (214 nm); retention time: 1.88 minutes (LC-MS Method 2).!H NMR (500 MHz, CD3OD) 58.5-7.7 (m, 1H), 7.89 (s, 1H), 7.61 (s, 1H), 7.16 (t, J = 7.5 Hz, 1H), 7.06 (s, 1H), 7.00 (d, J = 7.5 Hz, 2H), 6.55 (d, J = 7.0 Hz, 1H), 3.81 (s, 3H), 3.40-3.35 (m, 1H), 3.10-2.88 (m, 5H), 2.64-2.56 (m, 2H), 2.38-2.30 (m, 1H), 1.85 (t, 7= 12.0 Hz, 1H), 1.69 (s, 3H), 1.60 (s, 2H), 1.48-1.39 (m, 1H), 1.35-1.20 (m, 2H), 1.14 (d, J = 5.0 Hz, 6H), 1.06 (d, 7= 6.5 Hz, 3H) ppm.

[1777]

[0314] Example 42: Diastereomer 2, MS (ESI): 731 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.89 minutes (LC-MS Method 2). ’H NMR (500 MHz, CD3OD) 6 8.05-7.95 (m, 2H), 7.60 (t, 7 = 9.0 Hz, 1H). 7.14 (t, 7 = 8.0 Hz, 1H), 7.10-7.05 (m, 1H), 7.04-6.97 (m, 2H), 6.63-6.53 (m, 1H), 3.88-3.78 (m, 3H), 3.58-3.46 (m, 1H), 3.28-3.16 (m, 1H), 3.06-2.89 (m, 4H), 2.66-2.54 (m, 2H), 2.31 (t, 7 = 10.5 Hz, 1H), 1.96-1.82 (m, 1H), 1.69 (s, 3H), 1.64-1.54 (m, 1H), 1.45-1.30 (m, 2H), 1.25 (s, 3H), 1.16-1.01 (m, 8H) ppm.

[1778] Example 43: (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21,21-tetraoxo-12L6,21L6-dithia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1779]

[1780] Methyl (2S)-3-[ 3-[ (6R)-17-[bis( tert-butoxycarbonyl)amino ]- 18,19, 25-trifluoro-3, 6,10,10-tetramethyl-12,12,21,21-tetraoxo-12 6,21 6-dithia-3,4,27- 147

[1781] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1782] triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl ]phenyl ]-2-methyl-propanoate

[1783]

[1784]

[0315] Step A: To a stirred solution of methyl (2S)-3-[3-[(6R)-17-[bis(tert-butoxycarbonyl)amino]-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step A product of Example 38, 50 mg, 0.054 mmol) in dichloromethane (2 mL) was added meta-chloroperoxybenzoic acid (47 mg, 0.27 mmol). The reaction mixture was stirred at room temperature for 3 days, diluted with water (5 mL), and extracted with ethyl acetate (5 mL x 2). The combined organic extracts were washed with brine (5 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (20 g silica gel column, eluting with 0-60% ethyl acetate in petroleum ether) to give the title compound (40 mg, 76 %) as a yellow solid. MS (ESI): 961 m / z [M+H]+; LC purity: 95% (214 nm); retention time: 1.79 minutes (LC-MS Method 3). (2S)-3-[3-[(6R)-17-Amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21,21-tetraoxo- 12X6,2126-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoic acid

[1785]

[1786]

[0316] Step B: The title compound (11.4 mg, white solid) was prepared from Step A product (40 mg, 0.042 mmol) utilizing the reaction conditions described in Step E and Step F of Example 2, 3 and 4.

[1787]

[0317] MS (ESI): 747 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.94 minutes (LC-MS Method 2). ’H NMR (500 MHz, CD3OD) 5 8.38-8.26 (m, 1H), 8.19 (d, J = 4.0 Hz, 1H), 7.62 (t, J= 9.0 Hz, 1H), 7.15 (t, 7 = 7.5 Hz, 1H), 7.08 (s, 1H), 7.04-6.95 (m, 2H), 6.56 (d, 7 = 7.5 Hz, 1H), 3.84 (s, 3H), 3.46-3.35 (m. 3H), 3.05 (q, 7= 14.0 Hz, 2H), 2.94 (s, 1H), 2.60

[1788] 148

[1789] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1790] (s, 2H), 2.36 (t, 7 = 11.0 Hz, 1H), 1.88 (t, J = 11.0 Hz, 1H), 1.71 (s, 3H), 1.65-1.35 (m, 4H), 1.34-1.28 (m, 1H), 1.26 (s, 3H), 1.16 (s, 3H), 1.05 (s, 3H) ppm.

[1791] Example 44 and 45: Diastereomers 1 and 2 of methyl (2S)-3-[3-[(6R)-17-amino-16-chloro-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12X6,21X4-dithia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1792] F

[1793] , /

[1794]

[1795]

[0318] To a stirred solution of diastereomeric mixture of methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-12A6,21X4-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step A product of Example 41 and 42, 40 mg, 0.053 mmol) in acetic acid (4 mL) was added N-chlorosuccinimide (10.7 mg, 0.08 mmol). The mixture was stirred at 50 °C for 4.5 hours and concentrated. The residue was purified by prep-HPLC. The first eluent, Diastereomer 1, was designated as Example 44 ( 5.6 mg, 13%, a white solid). The second eluent, Diastereomer 2, was designated as Example 45 (5.8 mg, 13%, a white solid).

[1796]

[0319] Example 44: MS (ESI): 779 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.04 minutes (LC-MS Method 5).JH NMR (400 MHz, CD3OD) 5 8.66-7.92 (m, 2H), 7.60 (s, 1H), 7.16 (t, J= 8.0 Hz, 1H), 7.04-6.95 (m, 3H), 3.84 (s, 3H), 3.57 (s, 3H), 3.30-3.27 (m, 2H), 3.11-3.03 (m, 4H), 2.93-2.86 (m, 1H), 2.74-2.57 (m, 2H), 2.41 (s, 1H), 1.86 (t, J= 12.0 Hz, 1H), 1.71 (s, 3H), 1.12-1.06 (m, 13H) ppm.

[1797]

[0320] Example 45: MS (ESI): 779 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.10 minutes (LC-MS Method 5). ’H NMR (400 MHz, CD3OD) 5 8.19 (d, J= 4.0 Hz, 1H), 7.79 (s, 1H). 7.57 (t, J= 8.0 Hz, 1H), 7.16 (t, J= 8.0 Hz, 1H), 7.12-7.02 (m. 2H), 6.96 (d, J = 8.0 Hz, 1H), 3.89 (d, J = 2.8 Hz, 3H), 3.64-3.44 (m, 5H), 3.22-2.99 (m, 4H), 2.91-2.84 (m, 1H), 2.73-2.58 (m, 2H), 2.35-2.27 (m, 1H), 2.06-1.91 (m, 1H), 1.71 (s, 3H), 1.55-1.37 (m, 3H), 1.28-1.24 (m, 3H), 1.14 (s, 3H), 1.07 (d, J = 8.0 Hz, 4H) ppm.

[1798] Example 46: (2S)-3-[3-[(6R)-17-amino-16-chloro-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-

[1799] 149

[1800] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1801] triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1802]

[1803]

[0321] Utilizing the reaction conditions of Steps A described for Example 44 and 45, followed by that of Step I for Example 1, the title compound (8.7 mg, white solid) was prepared from methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step B product of Example 38, 197 mg, 0.27 mmol). MS (ESI): 749 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 2.02 minutes (LC-MS Method 5).JH NMR (500 MHz, CDsOD) 57.91 (dd, J = 6.5, 2.5 Hz, 1H), 7.81-7.78 (m, 1H), 7.34 (t, J = 9.0 Hz, 1H), 7.17 (t, J = 7.5 Hz, 1H), 7.07 (s, 1H), 7.01 (t, J = 8.0 Hz, 2H), 3.83 (d, J = 2.0 Hz, 3H), 3.67-3.48 (m, 2H), 3.20-3.07 (m, 2H), 3.03 (s, 2H), 2.95 (dd, J= 12.0, 5.5 Hz, 1H), 2.65-2.56 (m, 2H), 2.32 (t, J= 12.5 Hz, 1H), 1.95-1.90 (m, 1H), 1.70 (s, 3H), 1.61-1.45 (m. 3H), 1.28 (s, 1H), 1.21 (d, J= 5.5 Hz, 3H), 1.15 (s, 3H), 1.07 (d, J = 6.5 Hz, 3H) ppm.

[1804] Example 47: (2S)-3-[3-(17-amino-16-chloro-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21,21-tetraoxo-12X6,2U6-dithia-3,4,27-triazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl)phenyl]-2-methyl-propanoic acid

[1805]

[1806]

[0322] Utilizing the reaction conditions described in the preparation of Example 44 and 45, followed by Step D of Example 1, then Step A in Example 43, lastly Steps G and I of Example 1, the title compound (2.9 mg, white solid) was prepared from methyl (2S)-3-[3-[(6R)-17-amino-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12-dioxo-12X6,21-dithia-3,4,27-

[1807] 150

[1808] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1809] triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step B product of Example 38, 150 mg, 0.197 mmol). MS (ESI): 781 m / z. [M+H]+; LC purity: >99% (214 nm); retention time: 2.32 minutes (LC-MS Method 5). ’H NMR (500 MHz, CD3OD) 58.31 (dd, J= 6.5, 2.5 Hz, 1H), 8.28-8.23 (m, 1H), 7.64 (t, 7= 9.0 Hz, 1H), 7.16 (t, 7 = 7.5 Hz, 1H), 7.10 (s, 1H), 7.01 (dd, 7= 10.0, 4.5 Hz, 2H), 3.87 (d, 7 = 2.5 Hz, 3H), 3.60-3.53 (m, 1H), 3.49-3.42 (m, 2H), 3.27-3.20 (m, 1H), 3.17-3.14 (m, 1H), 3.09 (d, 7= 13.5 Hz, 1H), 3.00-2.91 (m, 1H), 2.65-2.56 (m, 2H), 2.39 (dd. 7 = 18.5, 11.5 Hz, 1H), 1.89 (dd, 7 = 19.0, 1.0 Hz, 1H), 1.73 (s, 3H), 1.62-1.54 (m, 2H), 1.51-1.45 (m, 1H), 1.36-1.32 (m, 1H), 1.28 (s, 3H), 1.22 (s, 3H), 1.06 (d, 7= 6.5 Hz, 3H) ppm.

[1810] Example 48: (2S)-3-[3-(17-amino-16-chloro-18,19,25-trifluoro-3,6,10,10-tetramethyl-12,12,21-trioxo-1216-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl)phenyl]-2-methyl-propanoic acid

[1811]

[1812]

[0323] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1, 1.30 g, 1.91 mmol) with the mixture of (4-(bis(tert-butoxycarbonyl)amino)-6-bromo-2,3-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide and (6-bromo-4-((tert-butoxycarbonyl)amino)-2,3-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-14, 2.78 g, 3.59 mmol) and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with (((S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoyl)oxy)methylium (Intermediate 9-2, 1.74 g, 3.59 mmol), the reaction procedure sequence, Steps A, B, C, D, E as described in Example 5, 6, 7, 8, followed by reaction conditions described in preparation of Example 44, 45, then Steps F and G of Example 5, 6, 7, 8, was used to give the title compound (1.6 mg, white solid). MS (ESI): 745 m z [M+H]+; LC purity: 95% (214 nm); retention time: 2.35 minutes (LC-MS Method 7). ’H NMR (500 MHz, CD3OD) 5 8.26-8.17 (m, 1H), 8.06 (dd, 7= 6.5, 2.0 Hz, 1H),

[1813] 151

[1814] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1815] 7.55 (t, 7= 9.0 Hz, 1H), 7.18-7.11 (m, 1H), 7.06-6.96(m, 3H), 3.86 (d, 7 = 2.0 Hz, 3H), 3.38-3.31 (m, 1H), 3.23- 3.07 (m, 3H), 2.92-2.84 (m, 2H), 2.63 -2.55 (m, 2H), 2.23 (t, 7= 12.5Hz, 1H), 1.94-1.83 (m, 1H), 1.69 (s, 3H), 1.56-1.55 (m, 1H), 1.44-1.24 (m, 3H), 1.07 (s, 3H), 1.04 (d, 7 = 6.5 Hz, 3H), 1.02-0.98 (m, 4H) ppm.

[1816] Example 49: (2S)-3-[3-[(6R)-17-Amino-16-chloro-19,25-difluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-1216-thia-3,4,27-triazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1817]

[1818]

[0324] Exchanging (3-(bis(tert-butoxycarbonyl)amino)-6-bromo-5-chloro-2,4-difluorophenyl)(4-fluoro-3-(imino(methylthio)methyl)phenyl)methyl acetate hydroiodide (Intermediate 11-1) with methyl 5-(4-amino-2-bromo-3-chloro-6-fluorophenoxy)-2-fluorobenzimidothioate hydroiodide (Intermediate 11-15, 5.30 g, 6.43 mmol), and methyl (R)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9) with methyl (S)-3-(3-((R)-7-((2-hydroxyethyl)sulfonyl)-2,6,6-trimethyl-l-(2-methylhydrazineyl)-l-oxoheptan-2-yl)phenyl)-2-methylpropanoate (Intermediate 9-2, 3.12 g, 6.43 mmol), the reaction procedure sequence, Steps A, B as described in Example 5, 6, 7 and 8, followed by Step D of Example 1, then Steps C, D, E and F of Example 5, 6, 7 and 8, was used to prepare the title compound (42 mg, 0.058 mmol) as a white solid. MS (ESI): 715 m / z [M+H]+; LC purity: >99% (214 nm); retention time: 1.99 minutes (LC-MS Method 5). ’H NMR (400 MHz, CD3OD) 5 7.36-7.28 (m, 2H), 7.23-7.14 (m, 2H), 7.05-7.00 (m, 3H), 6.69 (d, 7 = 12.0 Hz, 1H), 3.87 (d, 7= 1.6 Hz, 3H), 3.22-2.87 (m. 8H), 2.69-2.52 (m, 2H), 2.25 -2.18 (m, 1H), 1.91-1.80 (m, 1H), 1.73-1.61 (m. 4H), 1.47-1.28 (m, 2H), 1.13 (s, 6H), 1.06 (d, 7= 8.0 Hz, 3H) ppm.

[1819] Example 50: Methyl (2S)-3-[3-[(6R)-17-amino-25-fluoro-19-iodo-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-1216-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate

[1820] 152

[1821] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1822]

[1823]

[0325] To a stirred solution of methyl (2S)-3-[3-[(6R)-17-amino-25-fluoro-3,6,10,10-tetramethyl-12, 12-dioxo-21 -oxa- 12X6-thia-3,4, 18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Step G product of Example 1, 60 mg, 0.0885 mmol) in trifluoroacetic acid (1.0 mL, 13.0 mmol) was added N-iodosuccinimide (40 mg, 0.177 mmol). The mixture was stirred at room temperature for 16 hours, treated with saturated aqueous sodium bicarbonate (20 mL), and extracted with ethyl acetate (30 mL x 2). The combined organic extracts were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (20 g silica gel column, eluting with 0%-50% ethyl acetate in petroleum ether) to give the title compound (36 mg, 0.0448 mmol, 50.6 %). MS (ESI): 804 m / z [M+H]+; LC purity: 97% (214 nm); retention time: 1.76 minutes (LC-MS Method 2). ’H NMR (400 MHz, CD3OD) 8 7.72 (d, J = 8.4 Hz, 1H), 7.66 (s, 1H), 7.44-7.31 (m, 2H), 7.26 (dd, J = 4.8, 2.4 Hz, 1H), 7.00 (d, 7= 2.4 Hz, 1H), 6.84 (dd, J= 8.4, 2.4 Hz, 1H), 6.59 (s, 1H), 3.83 (d, J= 2.0 Hz, 3H), 3.55 (s, 3H), 3.26-3.17 (m, 2H), 3.07-2.87 (m, 5H), 2.80-2.69 (m, 2H), 2.22-2.12 (m, 1H), 1.86-1.76 (m, 1H), 1.67 (s, 3H), 1.62-1.53 (m, 1 H), 1.45-1.26 (m, 3H), 1.15-1.09 (m, 9H) ppm.

[1824] Example 51: (2S)-3-[3-[(6R)-17-amino-19-ethynyl-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12i6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoic acid

[1825]

[1826] Methyl (2S)-3-[3-[(6R)-17-cunino-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-19-(2-trimethylsilylethynyl)-21 -oxa-1276-thia-3, 4,18,27-

[1827] 153

[1828] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1829] tetrazatetracyclo[20.3.1.12, 5.015, 20]heptacosa-l(26), 2(27), 4, 15, 17, 19,22, 24-octaen-6-yl ]phenyl ]-2-methyl-propanoate

[1830] 'Si, F

[1831] /

[1832] ,. N

[1833]

[1834]

[0326] Step A: To a stirred solution of methyl (2S)-3-[3-[(6R)-17-amino-25-fluoro-19-iodo-3,6,10,10-tetramethyl-12,12-dioxo-21-oxa-12X6-thia-3,4,18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa-l(26),2(27),4,15,17,19,22,24-octaen-6-yl]phenyl]-2-methyl-propanoate (Example 50, 40 mg, 0.0498 mmol) in N, N-dimethylformamide (20 mL) were added bis(triphenylphosphine)palladium(ii) chloride (6.2 mg, 0.0085mmol), cuprous iodide (1.9 mg, 0.00995 mmol), triethylamine (0.02 mL, 0.15 mmol) and trimethylsilylacetylene (0.021 mL, 0.149 mmol). The mixture was stirred at 30 °C for 16 hours, diluted with 20 mL of water, and extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0-50% of ethyl acetate in petroleum ether) to give the title compound (30 mg, 0.0388 mmol, 78 %) as a yellow solid. MS (ESI): 774 m / z [M+H]+; LC purity: 99% (254 nm); retention time: 1.90 minutes (LC-MS Method 2).

[1835] Methyl ( 2S)-3-[ 3-[ ( 6R)-17-amino-19-elhynyl-25-fluoro-3,6, 10,10-tetramethyl-12, 12-dioxo- 21 -oxa- 12X6-thia-3,4, 18,27-tetrazatetracyclo[20.3.1.12,5.015,20]heptacosa- 1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl (phenyl ]-2-methyl-propanoate

[1836]

[1837]

[0327] Step B: To a stirred solution of Step A product (30 mg, 0.0388 mmol) in N, N-dimethylformamide (3 mL) was added cuprous iodide (1.5 mg, 0.00775 mmol). The mixture was stirred at 100 °C under Argon for 1 hour, diluted with 20 mL of water, and extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL), dried 154

[1838] IPTS / 200162816.3 Attorney Docket No. SLI-021WO

[1839] over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0-50% of ethyl acetate in petroleum ether) to give the title compound (15 mg, 0.0214 mmol, 55 %) as a yellow solid. MS (ESI): 702 m / z. [M+H]+; LC purity: 86% (254 nm); retention time: 1.70 minutes (LC-MS Method 2).

[1840] Example 51: (2S)-3-[3-[(6R)-17-Ammo-19-ethynyl-25-fluoro-3,6,10,10-tetramethyl-12,12-dioxo-21 -oxa-12X6-thia-3,4, 18,27 -tetr azatetracyclo[ 20.3.1.12,5.015,20]heptacosa-1(26), 2(27), 4, 15, 17, 19, 22, 24-octaen-6-yl ] phenyl ]-2-methyl-propanoic acid

[1841]

[1842]

[0328] Step C: To a stirred solution of Step B product (15 mg, 0.0214 mmol) in tetrahydrofuran / methanol / water mixture (1 mL / 1 mL / 1 mL) w...

Claims

Attorney Docket No. SLI-021WOClaims1. A compound of Formula I:Ior a pharmaceutically acceptable salt thereof,whereinL1is an optionally substituted Ci-6 alkylene chain wherein 1-3 of the methylene units is optionally and independently replaced by -O-, -N(R2)-, -C(O)-, -S-, -S(O)-, anoptionally substituted 3-6 membered carbocyclyl,, or optionally substituted C2 alkenylene;L2is an optionally substituted C1-6 alkylene chain wherein 1-3 of the methylene units is optionally and independently replaced by -C(CDs)2-, -O-, -N(R2), -C(O)-, -S-, -S(O)-, an optionally substituted 3-6 membered carbocyclyl, or optionally substituted C2 alkenylene;Ring A is an optionally substituted 5-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S;Ring C is optionally substituted phenyl or optionally substituted 5-6-membered heteroaryl comprising 1-4 heteroatoms each independently selected from the group consisting of N, O, and S;X is selected from the group consisting of -O-, -S-, -CH2-, -C(OH)H-, -S(O)-, -C(O)-, - 174IPTS / 200162816.3Attorney Docket No. SLI-021WOS(0)2-, -CFH-, -CF2-, and -N(R2)-;each RAis independently selected from the group consisting of halogen, cyano, optionally substituted Ci-Ce aliphatic, optionally substituted Ci-Ce alkoxy, and -CDs;each RBis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci-Ce aliphatic and optionally substituted Ci-Ce alkoxy;each Rcis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, and optionally substituted Ci -Co aliphatic; wherein at least one instance of Rcis -OR2or -N(R2)2;each RDis independently selected from the group consisting of halogen, cyano, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, optionally substituted Ci-Ce aliphatic, optionally substituted C1-C3 alkoxy, optionally substituted 5-6-membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S, wherein each RDis optionally substituted with 1-6 instances of Rd;wherein two instances of RDmay be taken together to form an optionally substituted 5-7 membered carbocyclic ring, optionally substituted 5-6- membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S; each Rdis independently selected from the group consisting of hydrogen, -OH, -CD3, - C(O)N(R2)2, -C(O)OR2, -OR2, -N(R2)2, -S(O)2R2, optionally substituted 5-6- membered heteroaryl, and optionally substituted 3-6-membered heterocyclyl comprising 1-3 heteroatoms selected from the group consisting of N, O, and S;R1is selected from the group consisting of hydrogen, cyano, -OR2, -(CH2)o-3N(R2)2, optionally substituted C1-C3 aliphatic, and -CD3;each R2is independently selected from hydrogen, optionally substituted Ci-Ce aliphatic, - OH, Ci-Ce alkoxy, -S(O)2(optionally substituted Ci-Ce aliphatic);n is 0, 1, 2, or 3;p is 0, 1, 2, 3, or 4;175IPTS / 200162816.3Attorney Docket No. SLI-021WOq is 1, or 2; andr is 0, 1, 2, 3, 4, or 5.

2. The compound of claim 1, wherein Ring A is optionally substituted 5-membered heteroaryl comprising 1-3 nitrogen atoms.

3. The compound of claim 1, wherein Ring A is optionally substituted 1,3,4-tnazolyl or optionally substituted imidazolyl.

4. The compound of claim 3, wherein Ring A is optionally substituted 1,3,4-triazolyl.N0N— N5. The compound of claim 4, wherein Ring A is6. The compound of claim 3, wherein Ring A is optionally substituted imidazolyl.N NA7. The compound of claim 6, wherein Ring A is 7^8. The compound of any claims 1-7, wherein Ring C is optionally substituted phenyl or optionally substituted 5-6-membered heteroaryl comprising 1-4 nitrogen atoms.

9. The compound of claim 8, wherein Ring C is phenyl.

10. The compound of claim 1-7, wherein Ring C is optionally substituted 6-membered heteroaryl comprising 1-4 nitrogen atoms.

11. The compound of claim 10, wherein Ring C is optionally substituted 6-membered heteroaryl comprising 1-2 nitrogen atoms.

12. The compound of claim 11, wherein Ring C is pyridine.

13. The compound of any claims 1-7, wherein the compound is selected from the group consisting of Formula (I-a), (I-b), (I-c), (I-d), (I-e), and (I-f):176IPTS / 200162816.3Attorney Docket No. SLI-021WO177IPTS / 200162816.3Attorney Docket No. SLI-021WOor a pharmaceutically acceptable salt thereof.

14. The compound of claim 13, wherein the compound is of formula (I-a-1) or (I-a-2)or a pharmaceutically acceptable salt thereof.

15. The compound of any claims 1-14, wherein each RAis independently optionally substituted Ci-Ce aliphatic or -CD3.

16. The compound of claim 15, wherein each RAis independently optionally substituted Ci-Ce aliphatic.

17. The compound of claim 16, wherein each RAis independently optionally substituted C1-C3 aliphatic.

18. The compound of claim 17, wherein each RAis independently optionally substituted C1-C2 aliphatic.

19. The compound of claim 18, wherein each RAis independently -CH3 or - CH2CH2N(H)CH3.

20. The compound of claim 19, wherein RAis -CH3.

21. The compound of any claims 1 -20, wherein RBis halogen.

22. The compound of claim 21, wherein RBis fluoro.

23. The compound of any claims 1-22, wherein each Rcis independently selected from the 178IPTS / 200162816.3Attorney Docket No. SLI-021WOgroup consisting of halogen, -OR2, -N(R2)2, and optionally substituted Ci-Ce aliphatic, wherein at least one instance of Rcis -OR2or -N(R2)2.

24. The compound of claim 23, wherein each Rcis independently halogen, -OR2or - N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2.

25. The compound of claim 24, wherein each Rcis independently selected from the group consisting of -F, -Cl, -I, -OR2, and -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2.

26. The compound of claim 25, wherein at least one instance of Rcis -F.

27. The compound of claim 23, wherein at least one instance of Rcis -OR2.

28. The compound of claim 23, wherein at least one instance of Rcis -N(R2)2.

29. The compound of claim 27, wherein at least one instance of Rcis -OH or -OCH.

30. The compound of claim 28, wherein at least one instance of Rcis -NH2 or -N(H)CH3.

31. The compound of claim 23, wherein Rcis optionally substituted CI-C aliphatic, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2.

32. The compound claim 31, wherein Rcis C1-C3 alkynyl, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2.

33. The compound of claim 32, Rcis -C=CH, -OR2or -N(R2)2, wherein at least one instance of Rcis -OR2or -N(R2)2.

34. The compound of any claims 1-33, wherein each RDis independently selected from the group consisting of halogen and C1-C6 aliphatic.

35. The compound of claim 34, wherein each RDis independently halogen.

36. The compound of claim 35, wherein RDis -F.

37. The compound of claim 34, wherein RDis optionally substituted C1-C6 aliphatic.

38. The compound of claim 37, wherein RDis optionally substituted C1-C4 aliphatic.179IPTS / 200162816.3Attorney Docket No. SLI-021WO39. The compound of claim 38, wherein at least one instance of RDis independentlyselected from the list consisting of CH3 CH3 CH3O OOH 'A V OH 40. The compound of claim 38, wherein RDis ^H3or<3H341. The compound of any claims 1-40, wherein R1is hydrogen, optionally substituted Ci- Ce aliphatic, or -CD3.

42. The compound of claim 41, wherein R1is optionally substituted Ci-Ce aliphatic.

43. The compound of claim 42, wherein R1is optionally substituted C1-C3 aliphatic.

44. The compound of claim 43, wherein R1is -CH3.

45. The compound of claim 41, wherein R1is -CD3.

46. The compound of any claims 1-45 wherein R2is hydrogen or optionally substituted Ci- C7 aliphatic.

47. The compound of claim 46, wherein R2is hydrogen.

48. The compound of claim 46, wherein R2is -CH3.

49. The compound of any claims 1-48, wherein each X is independently selected from the group consisting of -O-,-C(O)-, -S(O)-, -SO2-, -CH2-, -C(OH)H.

50. The compound of claim 49, wherein X is -O-.

51. The compound of claim 49, wherein X is -C(O)-.180IPTS / 200162816.3Attorney Docket No. SLI-021WO 52. The compound of claim 49, wherein X is -S(O)-.

53. The compound of claim 49, wherein X is -SO2-.

54. The compound of claim 49, wherein X is -CH2-.

55. The compound of claim 49, wherein X is -C(OH)H-.

56. The compound of any claims 1-55, wherein R1is optionally substituted C1-C6 aliphatic.

57. The compound of claim 56 wherein R1is optionally substituted C1-C3 aliphatic.

58. The compound of claim 57, wherein R1is -CH3.

59. The compound of any claims 1-58, wherein L1is an optionally substituted C1-6 alkylene.

60. The compound of claim 59, wherein L1is optionally substituted C1-3 alkylene.

61. The compound of claim 60, wherein L1is optionally substituted C2 alkylene.

62. The compound of claim 61, wherein L1is -CH2CH2-.

63. The compound of any claims 1-62, wherein L2is optionally substituted C1-C10 alkylene.

64. The compound of claim 63, wherein L2is optionally substituted C1-C6 alkylene.

65. The compound of claim 64, wherein L2is66. A compound selected from the group consisting of181IPTS / 200162816.3Attorney Docket No. SLI-021WO Example StructureNo.10 NZNo< A2X o oZ \, O3 / Z^_ z \oI o\o="X X y - Diastereomer 14Diastereomer 25 OxP VCl.H2N'FZ / ;N \0H°FDiastereomer 16rjJ VJXIJ^OH -^^40Diastereomer 2 F182IPTS / 200162816.3Attorney Docket No. SLI-021WO78F9 y 9., PFA J 1IIXXNH2N / F10 V 9>zpA JZ,'' hXXNH2N NN '1 — < y- >> os / 'f >0FZ11oTX12Diastereomer 113 °"° Vk n iFDiastereomer 2183IPTS / 200162816.3Attorney Docket No. SLI-021WO184IPTS / 200162816.3Attorney Docket No. SLI-021WO 21FifX^NH2AF22 F1OZZVA23 F24Diastereomer 125 RN'N^X-OHFHOzVW\ A^NH2w60 FDiastereomer 226.■ v;2728F185IPTS / 200162816.3Attorney Docket No. SLI-021WO Diastereomer 129FDiastereomer 230F OF31- T - F32 FF A^"rN'o< X33 F / i Lzv. y jV A4TA34 FFA "'°'o A35o “jA x'yA„.Diastereomer 1186IPTS / 200162816.3Attorney Docket No. SLI-021WO 36N-NT4) OHFWFDiastereomer 23738o'SoA39OZXV A40 F41o'& ADiastereomer 142XX^^rolxDiastereomer 243 Fp Q ftH2N'^AO'TA187IPTS / 200162816.3Attorney Docket No. SLI-021WO 44 FF O'iY " A. Jp'Cf i °O'T~ADiastereomer 145 FF O.O'T ADiastereomer 246 FF X^^\TN'H2FNI / yY\ " LAA-LMoz0 A47 E■N7rX) o _L48 oA vclxX rn =AAMkAyOH0F49 FO'I A50 F / H2NA^0OZV A51 F1 ° N AH2N J-^Q^\ / AY ir Y^oHo'b A188IPTS / 200162816.3Attorney Docket No. SLI-021WO 52O'T A53O" VA54 FO'I A55 FO'VA56I' - 0^™O'VA574TK58 F / X1< x59 FF o_x1H0X^OZT A189IPTS / 200162816.3Attorney Docket No. SLI-021WOor a pharmaceutically acceptable salt thereof.

67. A pharmaceutical composition comprising a compound of any of the previous claims and a pharmaceutically acceptable excipient.

68. A method of treating a CFTR-mediated disease or condition comprising administering a patient in need there of a compound any of claims 1-66 or a pharmaceutical composition of claim 67.

69. The method of claim 68, wherein the disease or condition is selected from cystic fibrosis, asthma, smoke induced COPD, chronic bronchitis, rhinosinusitis, constipation, pancreatitis, pancreatic insufficiency, male infertility caused by congenital bilateral absence of the vas deferens (CBAVD), mild pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, familial hypercholesterolemia, Type 1 chylomicronemia,190IPTS / 200162816.3Attorney Docket No. SLI-021WOabetalipoproteinemia, lysosomal storage diseases, I-cell disease / pseudo-Hurler, mucopolysaccharidoses, Sandhof / Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy / hyperinsulemia, Diabetes mellitus, Laron dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), neurophyseal DI, neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick’s disease, several polyglutamine neurological disorders, Huntington's, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, myotonic dystrophy, spongiform encephalopathies, hereditary Creutzfeldt- Jakob disease, Fabry disease, Straussler- Scheinker syndrome, COPD, dry-eye disease, Sjogren's disease, Osteoporosis, Osteopenia, bone healing and bone growth, bone repair, bone regeneration, reducing bone resorption, increasing bone deposition, Gorham's Syndrome, chloride channelopathies, myotonia congenita, Bartter's syndrome type III, Dent's disease, hyperekplexia, epilepsy, hyperekplexia, lysosomal storage disease, Angelman syndrome, Primary Ciliary Dyskinesia (PCD), PCD with situs inversus, PCD without situs inversus and ciliary aplasia.

70. The method of claim 68 or 69, wherein the disease or condition is selected from cystic fibrosis, congenital bilateral absence of vas deferens (CBAVD), acute, recurrent, or chronic pancreatitis, disseminated bronchiectasis, asthma, allergic pulmonary aspergillosis, chronic obstructive pulmonary disease (COPD), chronic sinusitis, dry eye disease, protein C deficiency, Abetalipoproteinemia, lysosomal storage disease, type 1 chylomicronemia, mild pulmonary disease, lipid processing deficiencies, type 1 hereditary angioedema, coagulation-fibrinolysis, hereditary hemochromatosis, CFTR- related metabolic syndrome, chronic bronchitis, constipation, pancreatic insufficiency, hereditary emphysema, and Sjogren's syndrome.

71. The method of any one of claims 68-70, wherein the disease or condition is cystic fibrosis.

72. A method of treating kidney disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound of any of claims 1-66 or a 191IPTS / 200162816.3Attorney Docket No. SLI-021WOpharmaceutical composition of claim 67.

73. The method of claim 72, wherein the kidney disease is autosomal dominant polycystic kidney disease or autosomal recessive polycystic kidney disease.

74. The method of claim 72, wherein the kidney disease is autosomal dominant polycystic kidney disease.

75. The method of claim 72, wherein the kidney disease is autosomal recessive polycystic kidney disease.

76. A method of treating cystic fibrosis in a subject, comprising administering to the subject a therapeutically effective amount of a compound of any of claims 1-66 or a pharmaceutical composition of claim 67.

77. The method of claim 76, wherein the subject is human.192IPTS / 200162816.3

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