Modulators of indoleamine 2,3-dioxygenase

Abstract

Provided are IDO inhibitor compounds of Formula I and pharmaceutically acceptable salts thereof, their pharmaceutical compositions, their methods of preparation, and methods for their use in the prevention and/or treatment of diseases. ##STR00001##

Claims

1. A compound of Formula II ##STR00122## or a pharmaceutically acceptable salt thereof, wherein in each depicted ring containing Xs, either each X is CH to form a benzene ring, or one X is N while the others are CH to form a pyridine ring; the depicted tetrazole may optionally be substituted by a CH.sub.2OH, wherein said CH.sub.2OH is optionally converted into a prodrug by converting the CH.sub.2OH group to a CH.sub.2OC(O)CH.sub.3, CH.sub.2OC(O)C(C.sub.1-4alkyl).sub.3, or OP(O)(OH).sub.2 group, or OP(O)(OC.sub.1-4alkyl).sub.2 group; R.sup.3 is phenyl, or a 4 to 6-membered heterocycle or heteroaryl containing 1 to 4 heteroatoms selected from N, S, and O; and wherein said phenyl, heterocycle, or heteroaryl, may optionally substituted by one to three C.sub.1-3alkyl groups each of which is optionally substituted by 1-3 halogens.

2. A pharmaceutical composition comprising a compound or salt according to claim 1.

Description

DETAILED DESCRIPTION OF REPRESENTATIVE EMBODIMENTS

(1) Preferably n is 0.

(2) Preferably, when one X is N, said N is positioned adjacent to the carbon atom to which the depicted S is bonded.

(3) Preferably, R.sup.1 is C.sub.1-6alkyl. Most preferably, R.sup.1 is t-butyl.

(4) Preferably, when A is a benzene ring or a pyridine ring, R.sup.2 is ortho to the depicted X-containing ring in Formula I. Preferably, when A is C.sub.1-4alkylene, A is ethylene or isopropylene.

(5) Preferably A is a benzene or pyridine ring. Most preferably A is a benzene ring.

(6) Preferably, R.sup.2 is a 5-membered heterocycle or heteroaryl containing 1 to 4 heteroatoms selected from N, S, and O, wherein said heterocycle or heteroaryl may optionally be substituted by a substituent selected from the group consisting of halogen, C.sub.3-6cycloalkyl, CH.sub.2OH, CH.sub.2OC.sub.1-3alkyl, C.sub.1-3alkyleneOC(O)C.sub.1-4alkyl, C.sub.1-3alkyl optionally substituted by 1-3 halogens, and wherein said CH.sub.2OH is optionally converted into a prodrug by converting the CH.sub.2OH group to a CH.sub.2OC(O)CH.sub.3, CH.sub.2OC(O)C(C.sub.1-4alkyl).sub.3, or OP(O)(OH).sub.2 group, or OP(O)(OC.sub.1-4alkyl).sub.2 group. Most preferably R.sup.2 is a tetrazole ring optionally substituted by CH.sub.2OH and wherein said CH.sub.2OH is optionally converted into a prodrug by converting the CH.sub.2OH group to a CH.sub.2OC(O)CH.sub.3, CH.sub.2OC(O)C(C.sub.1-4alkyl).sub.3, or OP(O)(OH).sub.2 group, or OP(O)(OC.sub.1-4alkyl).sub.2 group.

(7) Preferably L is NHC(O)C.sub.1-3alkylene. Most preferably L is NHC(O)CH.sub.2.

(8) Preferably R.sup.3 is phenyl, or a 4 to 6-membered heterocycle or heteroaryl containing 1 to 4 heteroatoms selected from N, S, and O; and wherein said phenyl, heterocycle, or heteroaryl, may optionally substituted by one to three substituents selected from the group consisting of halogen, cyano, C.sub.1-3alkyl optionally substituted by 1-3 halogens, C.sub.3-6cycloalkyl, SO.sub.2CH.sub.3, OCH.sub.3, CH.sub.2OH, C.sub.3-6cycloalkyl, CH.sub.2OC.sub.1-4alkyl, C(O)OC.sub.1-4alkyl, C(O)CH.sub.3, and OCH.sub.3, or two substituents bonded to adjacent atoms may join together to form a fused 5 or 6 membered ring optionally containing 1 or two heteroatoms selected from O, S, and N, fused to said C.sub.3-6cycloalkyl, phenyl, heterocycle, or heteroaryl. Most preferably, R.sup.3 is phenyl, or a 4 to 6-membered heterocycle or heteroaryl containing 1 to 4 heteroatoms selected from N, S, and O; and wherein said phenyl, heterocycle, or heteroaryl, may optionally substituted by one to three C.sub.1-3alkyl groups each of which is optionally substituted by 1-3 halogens.

(9) In particular, it is expected that the compounds and composition of this invention will be useful for prevention and/or treatment of HIV; including the prevention of the progression of AIDS and general immunosuppression. It is expected that in many cases such prevention and/or treatment will involve treating with the compounds of this invention in combination with at least one other drug thought to be useful for such prevention and/or treatment. For example, the IDO inhibitors of this invention may be used in combination with other immune therapies such as immune checkpoints (PD1, CTLA4, ICOS, etc.) and possibly in combination with growth factors or cytokine therapies (IL21, IL-7, etc.).

(10) Pharmaceutically acceptable salt refers to pharmaceutically acceptable salts derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium, and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, and oxalate. Suitable salts include those described in P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use; 2002.

(11) The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, pharmaceutically acceptable salts refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts of the present invention include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or ACN are preferred.

(12) The phrase pharmaceutically acceptable is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

(13) In another embodiment of the invention, there is provided a compound of Formula I, wherein the compound or salt of the compound is used in the manufacture of a medicament for use in the treatment immunosuppression in a human.

(14) In another embodiment of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable diluent and a therapeutically effective amount of a compound as defined in Formula I.

(15) In one embodiment, the pharmaceutical formulation containing a compound of Formula I or a salt thereof is a formulation adapted for parenteral administration. In another embodiment, the formulation is a long-acting parenteral formulation. In a further embodiment, the formulation is a nano-particle formulation.

(16) The present invention is directed to compounds, compositions and pharmaceutical compositions that have utility as novel treatments for immunosuppresion. While not wanting to be bound by any particular theory, it is thought that the present compounds are able to inhibit the enzyme that catalyzes the oxidative pyrrole ring cleavage reaction of I-Trp to N-formylkynurenine utilizing molecular oxygen or reactive oxygen species.

(17) Therefore, in another embodiment of the present invention, there is provided a method for the prevention and/or treatment of HIV; including the prevention of the progression of AIDS and general immunosuppression.

EXAMPLES

(18) The following examples serve to more fully describe the manner of making and using the above-described invention. It is understood that these examples in no way serve to limit the true scope of the invention, but rather are presented for illustrative purposes. In the examples and the synthetic schemes below, the following abbreviations have the following meanings. If an abbreviation is not defined, it has its generally accepted meaning. ACN=acetonitrile AIBN=azobisisobutyronitrile aq.=aqueous L or uL=microliters M or uM=micromolar NMR=nuclear magnetic resonance boc=tert-butoxycarbonyl br=broad Cbz=Benzyloxycarbonyl CDI=1,1-carbonyldiimidazole d=doublet =chemical shift C.=degrees celsius DCM=dichloromethane dd=doublet of doublets DHP=dihydropyran DIAD=diisopropyl azodicarboxylate DIEA or DIPEA=N,N-diisopropylethylamine DMAP=4-(dimethylamino)pyridine DMEM=Dulbeco's Modified Eagle's Medium DMF=dimethylformamide EtOAc=ethyl acetate h or hr=hours HATU=1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate HCV=hepatitus C virus HPLC=high performance liquid chromatography Hz=hertz IU=International Units IC.sub.50=inhibitory concentration at 50% inhibition J=coupling constant (given in Hz unless otherwise indicated) KHMDS=potassium bis(trimethylsilyl)amide LCMS=liquid chromatography-mass spectrometry m=multiplet M=molar M+H.sup.+=parent mass spectrum peak plus H.sup.+ MeOH=methanol mg=milligram min=minutes mL=milliliter mM=millimolar mmol=millimole MS=mass spectrum MTBE=methyl tert-butyl ether N=normal NFK=N-formylkynurenine NBS=N-bromosuccinimide nm=nanomolar NMP=N-methyl-2-pyrrolidone PE=petroleum ether ppm=parts per million q.s.=sufficient amount s=singlet RT=room temperature Rf=retardation factor sat.=saturated t=triplet TEA=triethylamine tetrakis=tetrakis(triphenylphosphine)palladium(0) TFA=trifluoroacetic acid TFAA=trifluoroacetic anhydride THF=tetrahydrofuran

(19) .sup.1H NMR spectra were recorded on a Bruker Ascend 400 spectrometer. Chemical shifts are expressed in parts per million (ppm, units). Coupling constants are in units of hertz (Hz). Splitting patterns describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad).

(20) The analytical low-resolution mass spectra (MS) were recorded on Waters ACQUITY UPLC with SQ Detectors using a Waters BEH C18, 2.150 mm, 1.7 m using a gradient elution method.

(21) Solvent A: 0.1% formic acid (FA) in water;

(22) Solvent B: 0.1% FA in acetonitrile;

(23) 30% B for 0.5 min followed by 30-100% B over 2.5 min.

(24) ##STR00003##

Example 1: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(25) ##STR00004##

Step A

(4-Bromo-2-nitrophenyl)(tert-butyl)sulfane

(26) ##STR00005##

(27) To a solution of 4-bromo-1-fluoro-2-nitrobenzene (5.53 g, 24.39 mmol) in acetonitrile (100 mL) was added cesium carbonate (7.23 g, 22.18 mmol) followed by 2-methylpropane-2-thiol (2.50 mL, 22.18 mmol) and the mixture was stirred at ambient temperature for 18 h and then at 70 C. for 5 h. The mixture was filtered washing the solid with EtOAc. The filtrate was concentrated and purified on silica gel (EtOAc/hexanes 0-5%) to provide (4-bromo-2-nitrophenyl)(tert-butyl)sulfane (6.01 g, 18.64 mmol, 84% yield) as a yellow oil. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.32 (s, 9H), 7.55-7.60 (m, 1H), 7.61-7.67 (m, 1H), 7.81 (d, J=1.95 Hz, 1H).

Step B

5-Bromo-2-(tert-butylthio)aniline

(28) ##STR00006##

(29) To a solution of (4-bromo-2-nitrophenyl)(tert-butyl)sulfane (6.01 g, 20.71 mmol) in ethanol (140 mL) was added zinc (12.31 g, 188 mmol), ammonium chloride (10.07 g, 188 mmol) and water (28.0 mL) and the mixt. was stirred at ambient temperature for 2 h. The mixture was filtered and concentrated and the residue was partitioned between EtOAc and water. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and dried to provide the desired product (4.97 g, 18.15 mmol, 96% yield) as a yellow solid. LCMS (M+H).sup.+: m/z=260.05. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.31 (s, 9H), 6.81 (dd, J=8.20, 2.15 Hz, 1H), 6.93 (d, J=1.95 Hz, 1H), 7.21 (d, J=8.20 Hz, 1H).

Step C

N-(5-Bromo-2-(tert-butylthio)phenyl)-2-(p-tolyl)acetamide

(30) ##STR00007##

(31) To a mixture of 5-bromo-2-(tert-butylthio)aniline (1.31 g, 4.78 mmol), 2-(p-tolyl)acetic acid (0.79 g, 5.26 mmol) and DIEA (2.08 mL, 11.96 mmol) in EtOAc (35 mL) was added 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (3.65 g, 5.74 mmol) (50%/EtOAc) and the mixture was stirred at ambient temperature for 18 h. The mixture was washed with water and the organic phase was dried (Na.sub.2SO.sub.4), concentrated and dried to provide the desired product (1.62 g, 4.13 mmol, 86% yield) as a yellow solid. LCMS (M+1).sup.+: m/z=392.2. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.01 (s, 9H), 2.38 (s, 3H), 3.76 (s, 2H), 7.12-7.20 (m, 1H), 7.28 (s, 6H), 8.83 (d, J=2.07 Hz, 1H).

Step D

N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(32) ##STR00008##

(33) A solution of N-(5-bromo-2-(tert-butylthio)phenyl)-2-(p-tolyl)acetamide (1.61 g, 4.10 mmol) in DMF (30 mL) was degassed with a stream of nitrogen while sequentially adding (2-(2H-tetrazol-5-yl)phenyl)boronic acid (2.34 g, 12.31 mmol), potassium carbonate (2.27 g, 16.41 mmol), water (6.00 mL) and tetrakis(triphenylphosphine) palladium(0) (0.47 g, 0.410 mmol) and then placed in a pre-heated oil bath at 100 C. The temperature was increased to 130 C. and the mixture was stirred under nitrogen atmosphere for 2 h. Water was added and 1N HCl/water was added to pH4-5. The solid was filtered washing with water. The solid was dissolved in EtOAc and the org. phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/dichloromethane 0-40%) to provide the title compound (1.32 g, 2.88 mmol, 70.3% yield) as a light pink solid. LCMS (M+1).sup.+: m/z=458.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.01 (s, 9H), 2.31 (s, 3H), 3.72 (s, 2H), 6.71 (dd, J=8.01, 1.76 Hz, 1H), 7.20-7.25 (m, 2H), 7.26-7.31 (m, 2H), 7.34 (d, J=7.81 Hz, 1H), 7.54-7.65 (m, 2H), 7.67-7.75 (m, 2H), 8.21 (s, 1H), 8.96 (s, 1H).

Examples 2 and 3: N-(4-(tert-butylsulfinyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide and N-(4-(tert-butylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(34) ##STR00009##

(35) To a solution of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (44 mg, 0.087 mmol) in acetic acid (0.5 mL)/dichloromethane (0.5 mL) was added dropwise a 3% solution of potassium permanganate (0.2 mL, 0.038 mmol). After 1 h at ambient temperature more 3% solution of potassium permanganate (0.1 mL, 0.019 mmol) was added. After 15 min saturated Na.sub.2S.sub.2O.sub.3/water was added followed by addition of saturated NaHCO.sub.3/water. The mixture was extracted with EtOAc and the organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified by HPLC (RP C18, MeCN/water 10-100%, 0.1% formic acid) to provide N-(4-(tert-butylsulfinyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (24.8 mg, 0.052 mmol, 60.5% yield) and N-(4-(tert-butylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (12.6 mg, 0.026 mmol, 29.7% yield). Example 2: LCMS (M+1).sup.+: m/z=474.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.98 (s, 9H), 2.28 (s, 3H), 3.55 (s, 2H), 6.94 (d, J=8.01 Hz, 1H), 7.10-7.25 (m, 4H), 7.38 (d, J=8.20 Hz, 1H), 7.53-7.66 (m, 2H), 7.67-7.78 (m, 3H), 10.41 (br. s., 1H). Example 3: LCMS (M+1).sup.+: m/z=490.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.09 (s, 9H), 2.29 (s, 3H), 3.69 (s, 2H), 6.96 (d, J=8.20 Hz, 1H), 7.13-7.26 (m, 4H), 7.56-7.69 (m, 3H), 7.70-7.82 (m, 2H), 8.31 (s, 1H), 9.78 (s, 1H).

Example 4: 4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxylic acid

(36) ##STR00010##

(37) This compound was prepared following the procedure described in Step D (Scheme I) from 2-boronobenzoic acid. LCMS (M+1).sup.+: m/z=434.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.04 (s, 9H), 2.31 (s, 3H), 3.75 (s, 2H), 7.03 (dd, J=7.91, 1.86 Hz, 1H), 7.21-7.26 (m, 2H), 7.29-7.34 (m, 2H), 7.38 (d, J=7.42 Hz, 1H), 7.44-7.52 (m, 2H), 7.56-7.63 (m, 1H), 7.75 (d, J=7.03 Hz, 1H), 8.35 (d, J=1.76 Hz, 1H), 9.01 (s, 1H), 12.80 (br. s., 1H).

Example 5: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(pyrazin-2-yl)acetamide

(38) ##STR00011##

(39) This compound was prepared from 2-(pyrazin-2-yl)acetic acid following the procedure described in Step C and D (Scheme I). LCMS (M+1).sup.+: m/z=446.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.16 (s, 9H), 4.05 (s, 2H), 6.76 (d, J=7.87 Hz, 1H), 7.40 (d, J=8.06 Hz, 1H), 7.59 (dd, J=18.86, 7.33 Hz, 2H), 7.66-7.75 (m, 2H), 8.09 (s, 1H), 8.61 (d, J=2.38 Hz, 1H), 8.67 (s, 1H), 8.73 (s, 1H), 9.64 (s, 1H).

Example 6: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(pyridin-4-yl)acetamide

(40) ##STR00012##

(41) This compound was prepared from 2-(pyridin-4-yl)acetic acid following the procedure described in Step C and D (Scheme I). LCMS (M+1).sup.+: m/z=445.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.10 (s, 9H), 3.84 (s, 2H), 6.77 (dd, J=8.06, 1.65 Hz, 1H), 7.39 (d, J=7.14 Hz, 3H), 7.53-7.64 (m, 2H), 7.69 (d, J=7.33 Hz, 2H), 8.01 (s, 1H), 8.56 (d, J=3.48 Hz, 2H), 9.23 (s, 1H).

Example 7: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-cyanophenyl)acetamide

(42) ##STR00013##

(43) This compound was prepared from 2-(4-cyanophenyl)acetic acid following the procedure described in Step C and D (Scheme I). LCMS (M+1).sup.+: m/z=469.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.09 (s, 9H), 3.91 (s, 2H), 6.77 (d, J=7.87 Hz, 1H), 7.39 (d, J=7.87 Hz, 1H), 7.52-7.64 (m, 4H), 7.69 (d, J=7.14 Hz, 2H), 7.86 (d, J=8.24 Hz, 2H), 8.02 (s, 1H), 9.18 (s, 1H).

Example 8: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(3-methylisoxazol-5-yl)acetamide

(44) ##STR00014##

(45) This compound was prepared from 2-(3-methylisoxazol-5-yl)acetic acid following the procedure described in Step C and D (Scheme I). LCMS (M+1).sup.+: m/z=449.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.18 (s, 9H), 2.24 (s, 3H), 4.05 (s, 2H), 6.34 (s, 1H), 6.80 (dd, J=8.01, 1.76 Hz, 1H), 7.43 (d, J=8.01 Hz, 1H), 7.55-7.67 (m, 2H), 7.70 (d, J=7.42 Hz, 2H), 8.00 (s, 1H), 9.39 (s, 1H).

Example 9: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(pyrimidin-5-yl)acetamide

(46) ##STR00015##

(47) This compound was prepared from 2-(pyrimidin-5-yl)acetic acid following the procedure described in Step C and D (Scheme I). LCMS (M+1).sup.+: m/z=446.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.16 (s, 9H), 3.89 (s, 2H), 6.79 (d, J=7.87 Hz, 1H), 7.42 (d, J=7.87 Hz, 1H), 7.58 (dd, J=19.59, 7.33 Hz, 3H), 7.69 (d, J=7.14 Hz, 2H), 7.93 (s, 1H), 8.79 (s, 2H), 9.11 (s, 1H), 9.42 (s, 1H).

Example 10: N-(4-(phenylthio)-2-(2H-tetrazol-5-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(48) ##STR00016##

(49) This compound was synthesized starting from 4-bromo-1-fluoro-2-nitrobenzene and thiophenol following Steps A-D (Scheme I) to provide the desired product as a trifluoroacetic acid salt. LCMS (M+1).sup.+: m/z=478.36. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 2.26 (s, 3H), 3.5 (br s, 1H), 3.57 (s, 2H), 6.82 (d, J=8.00 Hz, 1H), 7.02-7.10 (m, 4H), 7.12 (d, J=7.31 Hz, 2H), 7.19-7.39 (m, 4H), 7.51-7.75 (m, 5H), 9.37 (s, 1H).

(50) ##STR00017##

Example 11: N-(4-((3-methoxypropyl)thio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(51) ##STR00018##

Step A

(4-Bromo-2-nitrophenyl)(3-methoxypropyl)sulfane

(52) ##STR00019##

(53) A solution of 1-((3-methoxypropyl)sulfinyl)ethanone (0.81 g, 4.93 mmol) in EtOH (15 mL) was degassed with a stream of nitrogen for 10 min then potassium hydroxide (0.749 g, 11.34 mmol) (85%) was added and the mixture was stirred under nitrogen for 10 min until it became a cloudy solution. A solution of 4-bromo-1-fluoro-2-nitrobenzene (1.194 g, 5.43 mmol) in EtOH (1.5 mL) was added dropwise and the mixture was stirred at ambient temperature for 30 min. The mixture was concentrated and partitioned between EtOAc and water. The organic phase was washed with brine, dried (Na.sub.2SO.sub.4), concentrated to provide (4-bromo-2-nitrophenyl)(3-methoxypropyl)sulfane (1.3 g, 71% yield). .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.91-2.04 (m, 2H), 3.06 (t, J=7.32 Hz, 2H), 3.36 (s, 3H), 3.52 (t, J=5.76 Hz, 2H), 7.34 (d, J=8.79 Hz, 1H), 7.65 (dd, J=8.69, 2.25 Hz, 1H), 8.35 (d, J=2.15 Hz, 1H).

Step B-D

(N-(4-((3-methoxypropyl)thio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(54) ##STR00020##

(55) This compound was prepared from (4-bromo-2-nitrophenyl)(3-methoxypropyl)sulfane following Steps B-D described in Scheme I. LCMS (M+1).sup.+: m/z=474.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.66 (quin, J=6.64 Hz, 2H), 2.28 (s, 3H), 2.78 (t, J=7.13 Hz, 2H), 3.19 (s, 3H), 3.30-3.35 (m, 2H), 3.65 (s, 2H), 6.79 (d, J=7.62 Hz, 1H), 7.11-7.18 (m, 2H), 7.23 (d, J=7.62 Hz, 2H), 7.32 (d, J=8.20 Hz, 1H), 7.48-7.54 (m, 2H), 7.55-7.61 (m, 1H), 7.64-7.72 (m, 2H), 9.27 (s, 1H).

Example 12: N-(4-((tetrahydro-2H-pyran-4-yl)thio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(56) ##STR00021##

(57) This compound was synthesized starting from 4-bromo-1-fluoro-2-nitrobenzene and S-(tetrahydro-2H-pyran-4-yl) ethanethioate following the procedure described in Scheme II to provide the desired product. LCMS (M+1).sup.+: m/z=486.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.20-1.36 (m, 2H), 1.55 (d, J=12.11 Hz, 2H), 2.29 (s, 3H), 3.02 (t, J=10.55 Hz, 1H), 3.21 (t, J=10.55 Hz, 2H), 3.69 (s, 2H), 3.75 (d, J=11.52 Hz, 2H), 6.74 (d, J=7.03 Hz, 1H), 7.15-7.21 (m, 2H), 7.23-7.30 (m, 2H), 7.38 (d, J=8.01 Hz, 1H), 7.53 (d, J=7.62 Hz, 1H), 7.56-7.63 (m, 1H), 7.65-7.74 (m, 2H), 7.85 (s, 1H), 9.10 (s, 1H).

(58) ##STR00022##

Example 13: 1-(4-(tert-Butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-3-(p-tolyl)urea

(59) ##STR00023##

Step A

1-(5-Bromo-2-(tert-butylthio)phenyl)-3-(p-tolyl)urea

(60) ##STR00024##

(61) To a solution of 5-bromo-2-(tert-butylthio)aniline (200 mg, 0.715 mmol) in tetrahydrofuran (3 mL) was added 1-isocyanato-4-methylbenzene (143 mg, 1.072 mmol) and DIEA (0.2 mL). After 2 h stirring at ambient temperature, the mixture was heated to 70 C. for 8 h. More 1-isocyanato-4-methylbenzene (143 mg, 1.072 mmol) was added. After 18 h at 70 C. the mixture was concentrated and purified on silica gel (EtOAc/hexanes 0-5%) to provide the title compound (145 mg, 0.358 mmol, 50.0% yield) as a yellow foam. LCMS (M+1).sup.+: m/z=395.3. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.10 (s, 9H), 2.38 (s, 3H), 7.10 (dd, J=8.20, 2.15 Hz, 1H), 7.18-7.37 (m, 6H), 8.31 (s, 1H), 8.64 (d, J=2.15 Hz, 1H).

Step B

1-(4-(tert-Butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-3-(p-tolyl)urea

(62) ##STR00025##

(63) This compound was prepared from 1-(5-bromo-2-(tert-butylthio)phenyl)-3-(p-tolyl)urea and 2-(2H-tetrazol-5-yl)phenyl)boronic acid as described in Step D, Scheme I. LCMS (M+1).sup.+: m/z=459.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.27 (s, 9H), 2.24 (s, 3H), 6.61 (dd, J=7.87, 1.10 Hz, 1H), 7.09 (d, J=8.24 Hz, 2H), 7.29-7.39 (m, 3H), 7.61 (t, J=6.87 Hz, 2H), 7.66-7.75 (m, 2H), 8.18 (s, 1H), 8.50 (s, 1H), 9.61 (s, 1H).

Example 14: 4-(tert-Butylthio)-3-(3-(p-tolyl)ureido)-[1,1-biphenyl]-2-carboxylic Acid

(64) ##STR00026##

(65) This compound was prepared from 2-boronobenzoic acid following the procedure described in Scheme III. LCMS (M+1).sup.+: m/z=435.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.30 (s, 9H), 2.24 (s, 3H), 6.95 (dd, J=7.87, 2.01 Hz, 1H), 7.09 (d, J=8.24 Hz, 2H), 7.36 (d, J=8.42 Hz, 2H), 7.42 (d, J=7.69 Hz, 1H), 7.45-7.52 (m, 2H), 7.56-7.64 (m, 1H), 7.74 (d, J=7.69 Hz, 1H), 8.32 (d, J=1.83 Hz, 1H), 8.55 (s, 1H), 9.63 (s, 1H).

(66) ##STR00027##

Example 15: N-(4-(phenylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(67) ##STR00028##

Step A

N-(5-bromo-2-(phenylsulfonyl)phenyl)-2-(p-tolyl)acetamide

(68) ##STR00029##

(69) To a solution of N-(5-bromo-2-(phenylthio)phenyl)-2-(p-tolyl)acetamide (78 mg, 0.165 mmol) (synthesized from 4-bromo-1-fluoro-2-nitrobenzene and thiophenol following Steps A-C described for Scheme I) in acetic acid (1.00 mL)/dichloromethane (0.5 mL) was added dropwise a solution of potassium permanganate (867 mg, 0.165 mmol) (3%/water) and the mixture was stirred at ambient temperature for 18 h. More potassium permanganate (867 mg, 0.165 mmol) (3%/water) was added and stirring continued for 2 h. The mixture was diluted with EtOAc and water and solid NaHCO.sub.3 was carefully added. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-5-10%) to provide the title compound (60 mg, 0.132 mmol, 80% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=446.1. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 2.39 (s, 3H), 3.73 (s, 2H), 7.27-7.42 (m, 10H), 7.49-7.58 (m, 1H), 7.81 (d, J=8.53 Hz, 1H), 8.77 (d, J=1.79 Hz, 1H).

Step B

N-(4-(phenylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(70) ##STR00030##

(71) This compound was obtained from N-(5-Bromo-2-(phenylsulfonyl)phenyl)-2-(p-tolyl)acetamide and (2-(2H-tetrazol-5-yl)phenyl)boronic acid as described in Step D, Scheme I. LCMS: (M+1).sup.+: m/z=510.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 2.32 (s, 3H), 3.71 (s, 2H), 7.04 (d, J=8.56 Hz, 1H), 7.22 (s, 4H), 7.53 (t, J=7.69 Hz, 3H), 7.59-7.79 (m, 6H), 7.89-8.00 (m, 2H), 9.56 (s, 1H).

(72) ##STR00031##

Example 16: (N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(2,4-difluorophenyl)acetamide

(73) ##STR00032##

Step A

4-(tert-Butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine

(74) ##STR00033##

(75) A solution of 5-bromo-2-(tert-butylthio)aniline (760 mg, 2.92 mmol) in DMF (20 mL) was degassed with a stream of nitrogen while sequentially adding (2-(2H-tetrazol-5-yl)phenyl)boronic acid (1665 mg, 8.76 mmol), potassium carbonate (1615 mg, 11.68 mmol), water (4.00 mL) and tetrakis(triphenylphosphine) palladium(0) (338 mg, 0.292 mmol) and then placed in a pre-heated oil bath at 100 C. The temperature was increased to 130 C. and the mixture was stirred under nitrogen atmosphere for 1 h. Water was added and 1N HCl/water was added to pH-5. The mixture was extracted with EtOAc and the organic phase was washed with water. The organic phase was dried (Na.sub.2SO.sub.4), concentrated, and purified on silica gel (MeOH/dichloromethane 0-5%) to provide the title compound (1.05 g, 2.90 mmol, 99% yield). LCMS (M+1).sup.+: m/z=326.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.24 (s, 9H), 6.11 (dd, J=7.81, 1.76 Hz, 1H), 6.54 (d, J=1.76 Hz, 1H), 7.08 (d, J=7.81 Hz, 1H),7.2-7.3 (m, 1H), 7.50-7.59 (m, 2H), 7.60-7.72 (m, 2H), 7.95 (s, 1H).

Step B

N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(2,4-difluorophenyl)acetamide

(76) ##STR00034##

(77) To a solution of 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine (49 mg, 0.090 mmol) in EtOAc (1.5 mL) was added 2-(2,4-difluorophenyl)acetic acid (20.22 mg, 0.117 mmol), DIEA (0.047 mL, 0.271 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (74.7 mg, 0.117 mmol) (50%/EtOAc) and the mixture was stirred at ambient temperature for 18 h. The mixture was washed with water (1) and the organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified by HPLC (RP C18, MeCN/water. 0.1% formic acid) to provide the title compound (18.7 mg, 0.039 mmol, 43.2% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=480.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.12 (s, 9H), 3.82 (s, 2H), 6.74 (dd, J=8.01, 1.76 Hz, 1H), 7.15 (td, J=8.45, 2.25 Hz, 1H), 7.32 (td, J=9.81, 2.44 Hz, 1H), 7.38 (d, J=8.01 Hz, 1H), 7.49-7.64 (m, 3H), 7.66-7.74 (m, 2H), 8.11 (s, 1H), 9.13 (s, 1H).

Example 17: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-cyclopropylacetamide

(78) ##STR00035##

(79) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-cyclopropylacetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=408.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.28 (d, J=4.76 Hz, 2H), 0.55-0.67 (m, 2H), 0.98-1.12 (m, 1H), 1.25 (s, 9H), 2.32 (d, J=7.14 Hz, 2H), 6.75 (d, J=8.06 Hz, 1H), 7.42 (d, J=8.06 Hz, 1H), 7.55-7.66 (m, 2H), 7.70 (d, J=7.51 Hz, 2H), 8.22 (s, 1H), 9.35 (s, 1H).

Example 18: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-cyclohexylacetamide

(80) ##STR00036##

(81) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-cyclohexylacetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=450.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.90-1.05 (m, 2H), 1.07-1.30 (m, 12H), 1.54-1.83 (m, 6H), 2.27 (d, J=6.96 Hz, 2H), 6.76 (d, J=7.87 Hz, 1H), 7.41 (d, J=7.87 Hz, 1H), 7.60 (dd, J=11.08, 8.15 Hz, 2H), 7.66-7.78 (m, 2H), 8.01 (s, 1H), 9.08 (s, 1H).

Example 19: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(o-tolyl)acetamide

(82) ##STR00037##

(83) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(o-tolyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=458.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.99 (s, 9H), 2.25 (s, 3H), 3.78 (s, 2H), 6.73 (dd, J=7.91, 1.86 Hz, 1H), 7.22-7.29 (m, 3H), 7.31-7.42 (m, 2H), 7.55-7.66 (m, 2H), 7.70 (d, J=7.42 Hz, 2H), 8.23 (d, J=1.76 Hz, 1H), 8.85 (s, 1H).

Example 20: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(m-tolyl)acetamide

(84) ##STR00038##

(85) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(m-tolyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=458.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.01 (s, 9H), 2.32 (s, 3H), 3.73 (s, 2H), 6.71 (dd, J=7.91, 1.86 Hz, 1H), 7.12-7.23 (m, 3H), 7.27-7.32 (m, 1H), 7.34 (d, J=8.01 Hz, 1H), 7.53-7.65 (m, 2H), 7.67-7.76 (m, 2H), 8.20 (d, J=1.56 Hz, 1H), 8.98 (s, 1H).

Example 21: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(3-fluoro-4-methylphenyl)acetamide

(86) ##STR00039##

(87) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(3-fluoro-4-methylphenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=476.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.03 (s, 9H), 2.23 (s, 3H), 3.76 (s, 2H), 6.73 (dd, J=8.01, 1.76 Hz, 1H), 7.12 (d, J=7.62 Hz, 1H), 7.22 (d, J=10.74 Hz, 1H), 7.31 (t, J=7.91 Hz, 1H), 7.36 (d, J=8.01 Hz, 1H), 7.52-7.65 (m, 2H), 7.69 (d, J=7.42 Hz, 2H), 8.15 (d, J=1.37 Hz, 1H), 9.02 (s, 1H).

Example 22: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-methoxyphenyl)acetamide

(88) ##STR00040##

(89) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(4-methoxyphenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=474.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.01 (s, 9H), 3.70 (s, 2H), 3.75 (s, 3H), 6.71 (dd, J=8.01, 1.76 Hz, 1H), 6.98 (d, J=8.59 Hz, 2H), 7.27-7.38 (m, 3H), 7.53-7.65 (m, 2H), 7.69 (d, J=7.42 Hz, 2H), 8.21 (d, J=1.56 Hz, 1H), 8.95 (s, 1H).

Example 23: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(3-fluoro-4-methoxyphenyl)acetamide

(90) ##STR00041##

(91) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(3-fluoro-4-methoxyphenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=492.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.04 (s, 9H), 3.72 (s, 2H), 3.83 (s, 3H), 6.73 (dd, J=7.91, 1.86 Hz, 1H), 7.13-7.25 (m, 2H), 7.26-7.33 (m, 1H), 7.36 (d, J=7.81 Hz, 1H), 7.53-7.65 (m, 2H), 7.70 (d, J=7.42 Hz, 2H), 8.16 (s, 1H), 9.01 (s, 1H).

Example 24: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-fluorophenyl)acetamide

(92) ##STR00042##

(93) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(4-fluorophenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=462.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.05 (s, 9H), 3.78 (s, 2H), 6.73 (dd, J=7.91, 1.86 Hz, 1H), 7.23 (t, J=8.89 Hz, 2H), 7.36 (d, J=8.01 Hz, 1H), 7.44 (dd, J=8.50, 5.57 Hz, 2H), 7.53-7.64 (m, 2H), 7.66-7.76 (m, 2H), 8.13 (d, J=1.56 Hz, 1H), 9.04 (s, 1H).

Example 25: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-phenylacetamide

(94) ##STR00043##

(95) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-phenylacetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=444.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.02 (s, 9H), 3.78 (s, 2H), 6.72 (dd, J=8.01, 1.76 Hz, 1H), 7.35 (d, J=8.01 Hz, 2H), 7.37-7.46 (m, 4H), 7.54-7.65 (m, 2H), 7.69 (d, J=7.42 Hz, 2H), 8.17 (s, 1H), 9.02 (s, 1H).

Example 26: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(5-methylpyridin-2-yl)acetamide

(96) ##STR00044##

(97) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(5-methylpyridin-2-yl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=459.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.10 (s, 9H), 2.33 (s, 3H), 3.91 (s, 2H), 6.74 (d, J=8.01 Hz, 1H), 7.34-7.46 (m, 2H), 7.60 (dd, J=18.36, 7.23 Hz, 2H), 7.66-7.80 (m, 3H), 8.21 (s, 1H), 8.53 (s, 1H), 9.91 (s, 1H).

Example 27: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-isopropylphenyl)acetamide

(98) ##STR00045##

(99) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(4-isopropylphenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=486.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.98 (s, 9H), 1.20 (d, J=6.84 Hz, 6H), 2.89 (quin, J=6.84 Hz, 1H), 3.72 (s, 2H), 6.71 (d, J=7.81 Hz, 1H), 7.23-7.38 (m, 5H), 7.52-7.65 (m, 2H), 7.66-7.77 (m, 2H), 8.21 (s, 1H), 8.96 (s, 1H).

Example 28: N-(4-(tert-butylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-isopropylphenyl)acetamide

(100) ##STR00046##

(101) This compound was prepared from N-(4-(tert-butylsulfonyl)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-isopropylphenyl)acetamide following the procedure described in Example 2 and 3. LCMS (M+1).sup.+: m/z 518.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.03 (s, 9H), 1.20 (d, J=6.84 Hz, 6H), 2.75-2.90 (m, 1H), 3.70 (s, 2H), 6.95 (d, J=8.40 Hz, 1H), 7.24 (s, 4H), 7.61 (d, J=8.01 Hz, 2H), 7.67 (d, J=7.42 Hz, 1H), 7.75 (dd, J=19.14, 7.42 Hz, 2H), 8.34 (s, 1H), 9.78 (s, 1H).

Example 29: tert-Butyl 4-(2-((4-(tert-butylthio)-2-(2H-tetrazol-5-[1,1-biphenyl]-3-yl)amino)-2-oxoethyl)piperidine-1-carboxylate

(102) ##STR00047##

(103) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=551.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.98-1.14 (m, 2H), 1.23 (s, 9H), 1.35 (s, 9H), 1.62 (d, J=11.91 Hz, 2H), 1.91 (d, J=12.30 Hz, 1H), 2.34 (d, J=7.03 Hz, 2H), 2.45-2.55 (m, 2H), 3.89 (br. s., 2H), 6.78 (dd, J=7.81, 1.76 Hz, 1H), 7.42 (d, J=7.81 Hz, 1H), 7.61 (dd, J=9.67, 7.91 Hz, 2H), 7.67-7.77 (m, 2H), 7.95 (s, 1H), 9.15 (s, 1H), 12.09 (s, 1H).

Example 30: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(piperidin-4-yl)acetamide Hydrochloride

(104) ##STR00048##

(105) To tert-butyl 4-(2-((4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)amino)-2-oxoethyl)piperidine-1-carboxylate (26 mg, 0.042 mmol) (Example 29) was added HCl (0.499 mL, 1.997 mmol) (4M/dioxane) and the mixture was stirred at ambient temperature for 30 min. The mixture was concentrated and coevaporated with MeCN The residue was dissolved in dichloromethane and excess hexanes was added. The mixture was concentrated, dried to provide the title compound as the hydrochloride salt (21 mg, 0.042 mmol, 98% yield) as a yellowish solid. LCMS (M+1).sup.+: m/z=451.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.23 (s, 9H), 1.24-1.26 (m, 1H), 1.37 (d, J=12.69 Hz, 2H), 1.79 (d, J=13.87 Hz, 2H), 2.39 (d, J=7.03 Hz, 2H), 2.88 (d, J=10.16 Hz, 2H), 3.25 (d, J=12.30 Hz, 2H), 6.84 (dd, J=7.91, 1.86 Hz, 1H), 7.44 (d, J=8.01 Hz, 1H), 7.55-7.66 (m, 2H), 7.66-7.77 (m, 2H), 7.87 (s, 1H), 9.21 (s, 1H).

Example 31: 2-(1-Acetylpiperidin-4-yl)-N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)acetamide

(106) ##STR00049##

(107) To a suspension of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(piperidin-4-yl)acetamide (13 mg, 0.017 mmol) (Example 30) in dichloromethane (0.5 mL) was added TEA (0.014 mL, 0.104 mmol) followed by acetic anhydride (1.633 l, 0.017 mmol) and the mixture was stirred at ambient temperature for 15 min. The mixture was concentrated, dissolved in MeOH and purified by HPLC (RP C18, MeCN/water. 0.1% formic acid) to provide the title compound (3.3 mg, 6.43 mol, 37.1% yield) as a yellow solid. LCMS (M+1).sup.+: m/z=493.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.92-1.18 (m, 2H), 1.23 (s, 9H), 1.66 (t, J=15.04 Hz, 2H), 1.98 (s, 4H), 2.35 (d, J=7.23 Hz, 2H), 2.55-2.60 (m, 1H), 3.00 (t, J=11.72 Hz, 1H), 3.78 (d, J=13.47 Hz, 1H), 4.33 (d, J=13.28 Hz, 1H), 6.79 (dd, J=8.01, 1.76 Hz, 1H), 7.42 (d, J=7.81 Hz, 1H), 7.55-7.65 (m, 2H), 7.66-7.77 (m, 2H), 7.95 (s, 1H), 9.15 (s, 1H).

Example 32: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(1-(methylsulfonyl)piperidin-4-yl)acetamide

(108) ##STR00050##

(109) To a suspension of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(piperidin-4-yl)acetamide (63 mg, 0.084 mmol) (Example 30) in dichloromethane (1.5 mL) was added TEA (0.058 mL, 0.419 mmol) followed by methanesulfonyl chloride (0.013 mL, 0.168 mmol) and the mixture was stirred at ambient temperature for 15 min. The mixture was concentrated, dissolved in MeOH and purified by HPLC (RP C18, MeCN/water. 0.1% formic acid) to provide the title compound (15.7 mg, 0.030 mmol, 35.4% yield). LCMS (M+1).sup.+: m/z=529.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.23 (s, 9H), 1.23-1.33 (m, 3H), 1.75 (d, J=13.08 Hz, 2H), 2.39 (d, J=7.03 Hz, 2H), 2.63-2.77 (m, 2H), 2.85 (s, 3H), 3.53 (d, J=11.72 Hz, 2H), 6.79 (dd, J=7.91, 1.86 Hz, 1H), 7.43 (d, J=7.81 Hz, 1H), 7.61 (t, J=8.49 Hz, 2H), 7.70 (dd, J=7.03, 4.49 Hz, 2H), 7.94 (s, 1H), 9.18 (s, 1H).

Example 33: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-methoxyacetamide

(110) ##STR00051##

(111) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-methoxyacetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=398.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.26 (s, 9H), 3.47 (s, 3H), 4.03 (s, 2H), 6.79 (dd, J=7.91, 1.86 Hz, 1H), 7.45 (d, J=8.01 Hz, 1H), 7.56-7.66 (m, 2H), 7.67-7.77 (m, 2H), 8.30 (d, J=1.76 Hz, 1H), 9.83 (s, 1H).

Example 34: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(4-chlorophenyl)acetamide

(112) ##STR00052##

(113) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(4-chlorophenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=478.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.05 (s, 9H), 3.79 (s, 2H), 6.74 (d, J=7.62 Hz, 1H), 7.19 (d, J=8.20 Hz, 1H), 7.37 (dd, J=8.20, 2.15 Hz, 2H), 7.40-7.50 (m, 4H), 7.54-7.65 (m, 1H), 7.70 (d, J=7.42 Hz, 1H), 8.11 (s, 1H), 9.05 (s, 1H).

Example 35: p-Tolyl (4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)carbamate

(114) ##STR00053##

(115) To a solution of 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine (47 mg, 0.140 mmol) in dichloromethane (2 mL) was added TEA (0.064 mL, 0.462 mmol) and p-tolyl chloroformate (0.023 mL, 0.154 mmol) and the mixture was stirred at ambient temperature. After 1.5 h, more p-tolyl chloroformate (0.023 mL, 0.154 mmol) was added. After 18 h, the mixture was diluted with EtOAc and saturated NaHCO.sub.3/water and stirred at ambient temperature for 40 min; 1N HCl/water was added and the mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified by HPLC (RP C18, MeCN/water. 0.1% formic acid) to provide the title compound (17.2 mg, 0.037 mmol, 26.7% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=460.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.27 (s, 9H), 2.31 (s, 3H), 6.82 (d, J=8.06 Hz, 1H), 7.06 (d, J=8.24 Hz, 2H), 7.22 (d, J=8.24 Hz, 2H), 7.46 (d, J=7.87 Hz, 1H), 7.55-7.65 (m, 2H), 7.66-7.76 (m, 3H), 8.96 (s, 1H).

Example 36: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)benzamide

(116) ##STR00054##

(117) To a solution of 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine (49 mg, 0.146 mmol) in dichloromethane (2 mL) was added TEA (0.067 mL, 0.482 mmol) and benzoyl chloride (0.017 mL, 0.146 mmol) and the mixture was stirred at ambient temperature. After 1.5 h, more benzoyl chloride (0.017 mL, 0.146 mmol) was added. After 18 h, the mixture was diluted with EtOAc and saturated NaHCO.sub.3/water was added followed by 1M NaOH/water. After 1 h at ambient temperature the mixture was diluted with 1N HCl/water and extracted with EtOAc. The organic phase was concentrated and purified by HPLC (RP C18, MeCN/water, 0.1% formic acid) to provide the title compound (23.7 mg, 0.055 mmol, 37.8% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=430.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.23 (s, 9H), 6.86 (d, J=7.87 Hz, 1H), 7.50 (d, J=8.06 Hz, 1H), 7.57-7.68 (m, 5H), 7.72 (d, J=7.87 Hz, 2H), 7.94 (d, J=7.33 Hz, 2H), 8.15 (s, 1H), 9.83 (s, 1H).

Example 37: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-3-(p-tolyl)propanamide

(118) ##STR00055##

(119) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 3-(p-tolyl)propanoic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=472.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.17 (s, 9H), 2.25 (s, 3H), 2.66-2.75 (m, 2H), 2.78-2.89 (m, 2H), 6.75 (dd, J=7.97, 1.56 Hz, 1H), 7.03-7.10 (m, 2H), 7.10-7.16 (m, 2H), 7.39 (d, J=8.06 Hz, 1H), 7.59 (dd, J=17.31, 7.60 Hz, 2H), 7.69 (d, J=7.51 Hz, 2H), 8.02 (s, 1H), 9.11 (s, 1H).

Example 38: 2-(Benzo[d][1,3]dioxol-5-yl)-N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)acetamide

(120) ##STR00056##

(121) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(benzo[d][1,3]dioxol-5-yl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=488.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.05 (s, 9H), 3.68 (s, 2H), 6.02 (s, 2H), 6.72 (d, J=8.01 Hz, 1H), 6.82-6.90 (m, 1H), 6.92-6.97 (m, 1H), 7.00 (s, 1H), 7.35 (d, J=8.01 Hz, 1H), 7.52-7.65 (m, 2H), 7.70 (d, J=7.42 Hz, 2H), 8.20 (s, 1H), 8.99 (s, 1H).

Example 39: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-mesitylacetamide

(122) ##STR00057##

(123) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-mesitylacetic as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=486.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.93 (s, 9H), 2.25 (s, 9H), 3.72 (s, 2H), 6.69 (dd, J=7.81, 1.76 Hz, 1H), 6.97 (s, 2H), 7.31 (d, J=7.81 Hz, 1H), 7.54-7.65 (m, 2H), 7.70 (d, J=7.42 Hz, 2H), 8.30 (d, J=1.56 Hz, 1H), 8.72 (s, 1H).

Example 40: 2-(4-Bromophenyl)-N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)acetamide

(124) ##STR00058##

(125) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(4-bromophenyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=524.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.05 (s, 9H), 3.78 (s, 2H), 6.74 (dd, J=7.91, 1.86 Hz, 1H), 7.36 (dd, J=8.20, 2.54 Hz, 3H), 7.53-7.65 (m, 4H), 7.67-7.77 (m, 2H), 8.11 (d, J=1.56 Hz, 1H), 9.04 (s, 1H).

Example 41: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(2,4-dimethylphenyl)acetamide

(126) ##STR00059##

(127) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 2-(2,4-dimethylphenyl)acetic acid acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=472.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.97 (s, 9H), 2.21 (s, 3H), 2.28 (s, 3H), 3.72 (s, 2H), 6.72 (d, J=7.87 Hz, 1H), 7.03-7.14 (m, 2H), 7.25 (d, J=7.51 Hz, 1H), 7.33 (d, J=7.87 Hz, 1H), 7.59 (dd, J=17.58, 7.51 Hz, 2H), 7.69 (d, J=7.51 Hz, 2H), 8.25 (s, 1H), 8.79 (s, 1H).

Example 42: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-3-chloro-1,2,4-thiadiazol-5-amine

(128) ##STR00060##

(129) A solution of 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine (113 mg, 0.233 mmol) and 3,5-dichloro-1,2,4-thiadiazole (54.1 mg, 0.349 mmol) in DMSO (1 mL) was heated to 90 C. for 3 h. The mixture was purified by HPLC (C18 RP, MeCN/water 10-100%, 0.05% TFA) to provide the title compound (12 mg, 0.026 mmol, 11.39% yield) as a tan solid. LCMS (M+H).sup.+: m/z=444.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.23 (s, 9H), 6.83 (d, J=7.42 Hz, 1H), 7.48 (d, J=7.81 Hz, 1H), 7.65 (d, J=6.64 Hz, 2H), 7.73 (d, J=7.23 Hz, 2H), 8.15 (s, 1H), 10.79 (s, 1H).

Example 43: N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-3-(trifluoromethyl)-1,2,4-thiadiazol-5-amine

(130) ##STR00061##

(131) This compound was prepared from 4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-amine and 5-chloro-3-(trifluoromethyl)-1,2,4-thiadiazole as described in Example 42 to provide the title compound. LCMS: (M+1): m/z 478.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.24 (s, 9H) 6.80 (d, J=7.62 Hz, 1H) 7.48 (d, J=7.81 Hz, 1H) 7.59-7.68 (m, 2H) 7.74 (br. s., 2H) 8.28 (br. s., 1H) 10.95 (s, 1H).

(132) ##STR00062##

Example 44: 3-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)phenyl)butanoic Acid

(133) ##STR00063##

Step A

(E)-Ethyl 3-(4-(tert-butylthio)-3-nitrophenyl)but-2-enoate

(134) ##STR00064##

(135) To a solution of 5-bromo-2-(tert-butylthio)aniline (117 mg, 0.450 mmol) in DMF (2 mL) was added tetrabutylammonium bromide (29.0 mg, 0.090 mmol), ethyl crotonate (0.113 mL, 0.899 mmol), TEA (0.175 mL, 1.259 mmol) and bis(tri-o-tolylphosphine)palladium(II) dichloride (17.67 mg, 0.022 mmol) and the mixture was degassed with a stream of nitrogen for 5 min then heated under nitrogen at 110 C. for 1 h. Water was added and the mixture was extracted with EtOAc. The organic phase was washed with water, dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-5%) to provide the title compound (52.5 mg, 0.179 mmol, 39.8% yield) as a pale yellow oil. LCMS: (M+1): m/z=294.3. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.26-1.37 (m, 12H), 2.52 (d, J=0.98 Hz, 3H), 4.20 (q, J=7.03 Hz, 2H), 4.31-4.98 (m, 2H), 6.12 (d, J=1.17 Hz, 1H), 6.77 (dd, J=8.01, 1.95 Hz, 1H), 6.83 (d, J=1.76 Hz, 1H), 7.34 (d, J=8.01 Hz, 1H).

Step B

Ethyl 3-(3-amino-4-(tert-butylthio)phenyl)butanoate

(136) ##STR00065##

(137) To a solution of (E)-ethyl 3-(3-amino-4-(tert-butylthio)phenyl)but-2-enoate (50 mg, 0.170 mmol) in EtOH (2 mL) was added PdC (50 mg, 0.470 mmol) (10 wt. %, wet support, Degussa type E101 NEW) and the mixture was stirred at ambient temperature under hydrogen atmosphere for 2 h and then at 60 psi for 18 h. More PdC (50 mg, 0.470 mmol) was added and stirring at 60 psi of hydrogen continued for 24 h. The mixture was filtered and concentrated to provide the title compound (26.6 mg, 0.081 mmol, 47.6% yield). LCMS: (M+1): m/z=296.3. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.19 (t, J=7.05 Hz, 3H), 1.27 (d, J=7.14 Hz, 3H), 1.30 (s, 9H), 2.43-2.65 (m, 2H), 3.08-3.25 (m, 1H), 4.09 (q, J=7.14 Hz, 2H), 4.44 (br. s., 2H), 6.55 (d, J=7.87 Hz, 1H), 6.61 (d, J=1.47 Hz, 1H), 7.27 (d, J=7.87 Hz, 1H).

Step C

3-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)phenyl)butanoic Acid

(138) ##STR00066##

(139) To a solution of ethyl 3-(3-amino-4-(tert-butylthio)phenyl)butanoate (26 mg, 0.088 mmol) in EtOAc (1 mL) was added 2-(p-tolyl)acetic acid (13.22 mg, 0.088 mmol), DIEA (0.046 mL, 0.264 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (61.6 mg, 0.097 mmol) (50%/EtOAc) and the mixture was stirred at ambient temperature for 18 h. More 2-(p-tolyl)acetic acid (13.22 mg, 0.088 mmol), DIEA (0.046 mL, 0.264 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (61.6 mg, 0.097 mmol) (50%/EtOAc) was added. After 18 h, more DIEA (0.046 mL, 0.264 mmol), 2-(p-tolyl)acetic acid (13.22 mg, 0.088 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (61.6 mg, 0.097 mmol) (50%/EtOAc) was added. After 24 h the mixture was diluted with EtOAc and washed with water. The organic phase was dried (Na.sub.2SO.sub.4), concentrate and purified on silica gel (EtOAc/hexanes 0-10%) to provide ethyl 3-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)phenyl)butanoate (15 mg, 0.033 mmol, 37.5% yield) as a clear glass. This residue was dissolved in EtOH (0.5 mL) and 1M LiOH/water (0.13 mL) was added. After 18 h at ambient temperature the mixture was purified be HPLC (RP C18, MeCN/water 10-90%, 0.1% formic acid) to provide the title compound (11.6 mg, 0.029 mmol, 33.0% yield). LCMS: (M+1): m/z=400.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.99 (s, 9H), 1.20 (d, J=6.96 Hz, 3H), 2.30 (s, 3H), 2.46 (s, 2H), 3.12 (q, J=7.02 Hz, 1H), 3.73 (s, 2H), 6.99 (d, J=8.06 Hz, 1H), 7.18-7.25 (m, 2H), 7.26-7.32 (m, 2H), 7.35 (d, J=8.06 Hz, 1H), 8.24 (s, 1H), 8.92 (s, 1H), 12.2 (br s, 1H).

Example 45: 3-(4-(tert-Butylthio)-3-((3-chloro-1,2,4-thiadiazol-5-yl)amino)phenyl)butanoic Acid

(140) ##STR00067##

(141) A mixture of methyl 3-(3-amino-4-(tert-butylthio)phenyl)butanoate (98 mg, 0.313 mmol), 3,5-dichloro-1,2,4-thiadiazole (53.4 mg, 0.345 mmol) and DMF (1 mL) was heated at 90 C. for 5.5 h. water was added and the mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-10%) to provide methyl 3-(4-(tert-butylthio)-3-((3-chloro-1,2,4-thiadiazol-5-yl)amino)phenyl)butanoate (35.6 mg, 0.082 mmol, 26.1% yield). This residue was treated in MeOH (1 mL) and 1M LiOH/water (0.5 mL) and the mixture was stirred at ambient temperature for 30 min and then at 55 C. for 30 min. The mixture was purified by HPLC (C18 RP, MeCN/water 10-100%, 0.05% TFA) to provide the title compound (26.6 mg, 0.069 mmol, 21.99% yield) as a tan solid. LCMS (M+H).sup.+: m/z=386.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.21 (s, 9H), 1.25 (d, J=7.03 Hz, 3H), 2.52-2.55 (m, 2H), 3.18 (d, J=7.03 Hz, 1H), 7.10 (dd, J=8.01, 1.37 Hz, 1H), 7.49 (d, J=8.01 Hz, 1H), 8.12 (d, J=0.98 Hz, 1H), 10.74 (s, 1H), 12.12 (br. s., 1H).

(142) ##STR00068##

(143) A solution of 5-bromo-2-(tert-butylthio)aniline (206 mg, 0.752 mmol) in DMF (5 mL) was degassed with a stream of nitrogen while adding (2-cyanophenyl)boronic acid (133 mg, 0.903 mmol), potassium carbonate (312 mg, 2.256 mmol), water (1.000 mL) and tetrakis(triphenylphosphine) palladium(0) (87 mg, 0.075 mmol) and the mixture was placed in an oil bath preheated to 100 C. The temperature was increased to 110 C. After 2 h, water was added and the mixture was extracted with EtOAc and the organic phase was washed with water, brine, dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 10-100%) to provide the 3-amino-4-(tert-butylthio)-[1,1-biphenyl]-2-carbonitrile (57.4 mg, 0.201 mmol, 26.8% yield, LCMS (M+H).sup.+: m/z=283.1) and 3-amino-4-(tert-butylthio)-[1,1-biphenyl]-2-carboxamide (86 mg, 0.278 mmol, 36.9% yield, LCMS (M+H).sup.+: m/z=301.2).

Example 46: N-(4-(tert-butylthio)-2-cyano-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(144) ##STR00069##

(145) This compound was prepared from 3-amino-4-(tert-butylthio)-[1,1-biphenyl]-2-carbonitrile and 2-(p-tolyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=415.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.06 (s, 9H), 2.31 (s, 3H), 3.34 (s, 2H), 7.20-7.26 (m, 2H), 7.27-7.37 (m, 3H), 7.62 (t, J=7.81 Hz, 3H), 7.78-7.85 (m, 1H), 7.98 (d, J=7.23 Hz, 1H), 8.52 (d, J=1.76 Hz, 1H), 9.11 (s, 1H).

Example 47: 4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxamide

(146) ##STR00070##

(147) This compound was prepared from 3-amino-4-(tert-butylthio)-[1,1-biphenyl]-2-carboxamide and 2-(p-tolyl)acetic acid as described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=433.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.04 (s, 9H), 2.31 (s, 3H), 3.75 (s, 2H), 7.11 (dd, J=7.91, 1.86 Hz, 1H), 7.20-7.26 (m, 2H), 7.28-7.38 (m, 4H), 7.39-7.55 (m, 4H), 7.71 (s, 1H) 8.41 (d, J=1.76 Hz, 1H), 9.01 (s, 1H).

Example 48: N-(4-(tert-butylthio)-2-(N-hydroxycarbamimidoyl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(148) ##STR00071##

(149) To a suspension of hydroxylamine hydrochloride (46.1 mg, 0.663 mmol) in DMSO (1 mL) was added TEA (0.092 mL, 0.663 mmol) and the mixture was stirred at ambient temperature for 15 min. The solid was filtered and washed with THF (2 mL). The filtrate was concentrated to 1 mL and then added to N-(4-(tert-butylthio)-2-cyano-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (55 mg, 0.133 mmol). The mixture was stirred at 65 C. for 1 h and then at 75 C. for 48 h. The mixture was purified by HPLC (RP C18, MeCN/water 10-100%, 0.1% formic acid) to provide the title compound (8 mg, 0.018 mmol, 13.34% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=448.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.04 (s, 9H), 2.31 (s, 3H), 3.74 (s, 2H), 5.59 (s, 2H), 7.13 (d, J=7.62 Hz, 1H), 7.20-7.26 (m, 2H), 7.28-7.36 (m, 3H), 7.38-7.53 (m, 4H), 8.37 (s, 1H), 9.00 (s, 1H), 9.22 (s, 1H).

(150) ##STR00072##

Examples 49 and 50: Di-tert-butyl((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl)phosphate and Di-tert-butyl((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]H-tetrazol-1-yl)methyl)phosphate

(151) ##STR00073##

(152) To a solution of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (320 mg, 0.699 mmol) in DMF (4 mL) was added cesium carbonate (273 mg, 0.839 mmol), sodium iodide (105 mg, 0.699 mmol) and di-tert-butyl (chloromethyl) phosphate (0.197 mL, 0.839 mmol) and the mixture was stirred at 55 C. for 18 h. Water was added followed by 1N HCl/water to pH-4-5. The mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-25%) to provide di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl) phosphate (Example 49, 162 mg, 0.238 mmol, 34.1% yield) and di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl) phosphate (Example 50, 68 mg, 0.089 mmol, 12.73% yield). Example 49: LCMS (M+1).sup.+: m/z=680.6. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.04 (s, 9H), 1.45 (s, 18H), 2.37 (s, 3H), 3.70 (s, 2H), 6.16 (d, J=10.62 Hz, 2H), 6.80 (dd, J=7.87, 1.65 Hz, 1H), 7.22-7.28 (m, 4H), 7.30 (d, J=7.87 Hz, 1H), 7.45-7.60 (m, 3H), 7.88 (d, J=7.69 Hz, 1H), 8.41 (s, 1H), 8.77 (s, 1H). Example 50: LCMS (M+1).sup.+: m/z=680.6. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 0.99 (s, 9H), 1.38 (s, 18H), 2.37 (s, 3H), 3.72 (s, 2H), 5.60 (d, J=10.26 Hz, 2H), 6.66 (dd, J=7.87, 1.65 Hz, 1H), 7.26-7.33 (m, 5H), 7.56 (d, J=6.96 Hz, 1H), 7.61-7.72 (m, 3H), 8.48 (d, J=1.65 Hz, 1H), 8.80 (s, 1H).

Example 51: (5-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl Dihydrogen Phosphate

(153) ##STR00074##

(154) To a solution of di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl) phosphate (135 mg, 0.199 mmol) in dichloromethane (1 mL) was added TFA (1 ml, 12.98 mmol) and the mixture was stirred at ambient temperature for 4 h. The mixture was concentrated and coevaporated with MeCN and dried in vacuo. Hexanes was added, concentrated and dried in vacuo to provide the title compound as Trifluoroacetic acid salt (125.8 mg, 0.183 mmol, 92% yield) as a tan solid. LCMS (M+1).sup.+: m/z=568.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.02 (s, 9H), 2.31 (s, 3H), 3.71 (s, 2H), 6.15 (d, J=11.17 Hz, 2H), 6.80 (d, J=7.87 Hz, 1H), 7.19-7.25 (m, 2H), 7.26-7.31 (m, 2H), 7.35 (d, J=8.06 Hz, 1H), 7.54 (d, J=7.51 Hz, 1H), 7.57-7.63 (m, 1H), 7.67 (d, J=7.51 Hz, 1H), 7.81 (d, J=7.51 Hz, 1H), 8.11 (s, 1H), 8.95 (s, 1H).

Example 52: (5-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl Dihydrogen

(155) ##STR00075##

(156) A solution of HCl (0.5 mL, 2.000 mmol) (4M/dioxane) was added to di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl) phosphate (6 mg, 8.38 mol) and the mixture was stirred at ambient temperature for 30 min. The mixture was concentrated, coevaporated with MeCN, triturated with dichloromethane and excess hexanes, concentrated and dried in vacuo to provide the title compound (5 mg, 8.37 mol, 100% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=568.3. .sup.1H NMR (400 MHz, METHANOL-d.sub.4): ppm 0.95 (s, 9H), 2.33 (s, 3H), 3.69 (s, 2H), 5.85 (d, J=10.35 Hz, 2H), 6.88 (dd, J=7.81, 1.56 Hz, 1H), 7.18-7.30 (m, 4H), 7.37 (d, J=7.81 Hz, 1H), 7.62 (d, J=7.42 Hz, 1H), 7.67 (d, J=7.42 Hz, 2H), 7.73 (d, J=7.42 Hz, 1H), 8.14 (s, 1H).

Example 53: (5-(4-(tert-Butylthio)-3-(2-(4-isopropylphenyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl Dihydrogen Phosphate

(157) ##STR00076##

(158) To a solution of di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(4-isopropylphenyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl) phosphate (260 mg, 0.331 mmol prepared as described in Scheme VIII) in dichloromethane (2 mL) at 0 C. was added TFA (2 mL) dropwise and the mixture was stirred at ambient temperature for 3 h. The mixture was concentrated, coevaporated with MeCN, dried and a small amount of EtOAc was added. The solid was filtered and dried to provide the title compound (132 mg, 0.219 mmol, 66.4% yield) as an off-white solid. LCMS (M+1).sup.+: m/z=596.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.99 (s, 9H), 1.21 (d, J=7.03 Hz, 6H), 2.82-2.96 (m, 1H), 3.71 (s, 2H), 6.16 (d, J=11.13 Hz, 2H), 6.80 (d, J=6.25 Hz, 1H), 7.24-7.38 (m, 5H), 7.54 (d, J=7.62 Hz, 1H), 7.61 (d, J=7.42 Hz, 1H), 7.67 (d, J=7.42 Hz, 1H), 7.81 (d, J=7.42 Hz, 1H), 8.12 (s, 1H), 8.96 (s, 1H).

Example 54: (5-(4-(tert-Butylthio)-3-(2-(4-isopropylphenyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl Dihydrogen Phosphate

(159) ##STR00077##

(160) A mixture of di-tert-butyl ((5-(4-(tert-butylthio)-3-(2-(4-isopropylphenyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl) phosphate (72 mg, 0.102 mmol prepared as described in Scheme VIII) acetone (1.5 mL) and water (1.500 mL) was heated at 60 C. for 1 h. The mixture was concentrated and coevaporated with MeCN. The residue was dried, triturated with hexanes and the solid was filtered to provide the title compound (40.3 mg, 0.067 mmol, 65.9% yield) as a pale yellow solid. LCMS (M+1).sup.+: m/z=596.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.95 (s, 9H), 1.20 (d, J=6.84 Hz, 6H), 2.82-2.96 (m, 1H), 3.74 (s, 2H), 5.82 (d, J=10.35 Hz, 2H), 6.73 (d, J=7.62 Hz, 1H), 7.23-7.38 (m, 5H), 7.61-7.72 (m, 3H), 7.74-7.85 (m, 1H), 8.17 (s, 1H), 8.95 (s, 1H).

Examples 55 and 56: (5-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl pivalate and (5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl Pivalate

(161) ##STR00078##

(162) To a suspension of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (46 mg, 0.101 mmol) in acetonitrile (2 mL) was added potassium carbonate (34 mg, 0.246 mmol) followed by chloromethyl pivalate (0.016 mL, 0.111 mmol) and the mixture was stirred at 70 C. for 4 h. More chloromethyl pivalate (0.10 mL) was added and stirring at 70 C. continued for 5 h. Water was added and the mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-20%) to provide (5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl pivalate (23.5 mg, 0.041 mmol, 40.9% yield) and (5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl pivalate (21 mg, 0.037 mmol, 36.5% yield). Example 55: LCMS (M+1).sup.+: m/z=572.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.01 (s, 9H), 1.09 (s, 9H), 2.31 (s, 3H), 3.70 (s, 2H), 6.47 (s, 2H), 6.78 (dd, J=8.01, 1.95 Hz, 1H), 7.20-7.24 (m, 2H), 7.26-7.30 (m, 2H), 7.32 (d, J=8.01 Hz, 1H), 7.52 (d, J=7.42 Hz, 1H), 7.56-7.63 (m, 1H), 7.64-7.72 (m, 1H), 7.79 (d, J=7.62 Hz, 1H), 8.13 (d, J=1.56 Hz, 1H), 8.92 (s, 1H). Example 56: LCMS (M+1).sup.+: m/z=572.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.92-1.05 (m, 18H), 2.30 (s, 3H), 3.72 (s, 2H), 6.00 (s, 2H), 6.75 (dd, J=7.81, 1.95 Hz, 1H), 7.18-7.24 (m, 2H), 7.25-7.31 (m, 2H), 7.34 (d, J=8.01 Hz, 1H), 7.60-7.73 (m, 3H), 7.74-7.83 (m, 1H), 8.18 (d, J=1.76 Hz, 1H), 8.94 (s, 1H).

Examples 57 and 58: (5-(4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl Acetate and (5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl Acetate

(163) ##STR00079##

(164) To a suspension of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (83 mg, 0.163 mmol) in acetonitrile (1.5 mL) was added potassium carbonate (45.1 mg, 0.326 mmol) followed by bromomethyl acetate (0.024 mL, 0.245 mmol) and the mixture was stirred at 60 C. under nitrogen atmosphere for 30 min. Saturated NH.sub.4Cl/water and water was added and the mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-40%) to provide(5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-2H-tetrazol-2-yl)methyl acetate (31.3 mg, 0.059 mmol, 36.2% yield) and (5-(4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-yl)-1H-tetrazol-1-yl)methyl acetate (25.2 mg, 0.046 mmol, 28.0% yield). Example 57: LCMS (M+1).sup.+: m/z=530.4. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.04 (s, 9H), 2.12 (s, 3H), 2.37 (s, 3H), 3.70 (s, 2H), 6.35 (s, 2H), 6.82 (d, J=7.87 Hz, 1H), 7.21-7.29 (m, 4H), 7.33 (d, J=7.87 Hz, 1H), 7.45-7.62 (m, 3H), 7.89 (d, J=7.51 Hz, 1H), 8.37 (s, 1H), 8.76 (s, 1H). Example 58: LCMS (M+1).sup.+: m/z=530.5. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.00 (s, 9H), 1.95 (s, 3H), 2.38 (s, 3H), 3.73 (s, 2H), 5.70 (s, 2H), 6.63 (d, J=7.87 Hz, 1H), 7.21-7.34 (m, 5H), 7.52-7.61 (m, 2H), 7.62-7.76 (m, 2H), 8.54 (s, 1H), 8.82 (br. s., 1H).

Examples 59 and 60: N-(4-(tert-butylthio)-2-(2-(methoxymethyl)-2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide and N-(4-(tert-butylthio)-2-(1-(methoxymethyl)-1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(165) ##STR00080##

(166) To a suspension of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (55 mg, 0.113 mmol) in acetonitrile (1.2 mL) was added TEA (0.063 mL, 0.452 mmol), sodium iodide (15 mg, 0.100 mmol) and MOM-Cl (0.017 mL, 0.226 mmol) and the mixture was stirred at ambient temperature for 10 min. The mixture was partitioned between EtOAc and water. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-40%) to provide N-(4-(tert-butylthio)-2-(2-(methoxymethyl)-2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (19.8 mg, 0.037 mmol, 33.2% yield) and N-(4-(tert-butylthio)-2-(1-(methoxymethyl)-1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (24.2 mg, 0.048 mmol, 42.7% yield). Example 59: LCMS (M+1).sup.+: m/z=502.4. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.03 (s, 9H), 2.37 (s, 3H), 3.33 (s, 3H), 3.69 (s, 2H), 5.71 (s, 2H), 6.80 (dd, J=8.01, 1.95 Hz, 1H), 7.31 (d, J=8.01 Hz, 1H), 7.24-7.26 (m, 4H), 7.45-7.60 (m, 3H), 7.90 (d, J=7.62 Hz, 1H), 8.46 (d, J=1.76 Hz, 1H), 8.78 (s, 1H). Example 60: LCMS (M+1).sup.+: m/z=502.4. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 0.98 (s, 9H), 2.37 (s, 3H), 3.24 (s, 3H), 3.72 (s, 2H), 5.20 (s, 2H), 6.67 (dd, J=7.91, 1.86 Hz, 1H), 7.24-7.26 (m, 5H), 7.50-7.60 (m, 2H), 7.61-7.74 (m, 2H), 8.48 (d, J=1.76 Hz, 1H), 8.80 (s, 1H).

Examples 61 and 62: N-(4-(tert-butylthio)-2-(2-methyl-2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide and N-(4-(tert-butylthio)-2-(1-methyl-1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(167) ##STR00081##

(168) To a solution of N-(4-(tert-butylthio)-2-(2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (48 mg, 0.105 mmol) in THF (1 mL) was added TMS-diazomethane (0.5 mL, 1.000 mmol) (2M/hexanes) and the mixture was stirred at ambient temperature for 1 h. Water was added followed by a small amount of MeOH and excess EtOAc and the mixture was stirred at ambient temperature for 30 min. The mixture was washed with saturated NaHCO.sub.3/water and the organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-50%) to provide N-(4-(tert-butylthio)-2-(1-methyl-1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (15.4 mg, 0.033 mmol, 31.1% yield) and N-(4-(tert-butylthio)-2-(2-methyl-2H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (6.6 mg, 0.013 mmol, 12.67% yield). Example 61: LCMS (M+1).sup.+: m/z=472.4. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 0.99 (s, 9H), 2.38 (s, 3H), 3.40 (s, 3H), 3.75 (s, 2H), 6.53 (d, J=7.81 Hz, 1H), 7.24-7.27 (m, 5H), 7.50-7.62 (m, 2H), 7.66 (s, 2H), 8.66 (s, 1H), 8.86 (s, 1H). Example 62: LCMS (M+1).sup.+: m/z=472.4. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.03 (s, 9H), 2.37 (s, 3H), 3.70 (s, 2H), 4.23 (s, 3H), 6.80 (dd, J=8.01, 1.95 Hz, 1H), 7.24-7.26 (m, 4H), 7.31 (d, J=8.01 Hz, 1H), 7.44-7.59 (m, 3H), 7.85 (d, J=7.42 Hz, 1H), 8.41 (d, J=1.76 Hz, 1H), 8.77 (s, 1H).

(169) ##STR00082##

Example 63: N-(5-(2-(2H-tetrazol-5-yl)pyridin-3-yl)-2-(tert-butylthio)phenyl)-2-(p-tolyl)acetamide

(170) ##STR00083##

Step A

3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)picolinonitrile

(171) ##STR00084##

(172) A mixture of (2-cyanopyridin-3-yl)boronic acid (0.43 g, 2.91 mmol), toluene (12 mL), pinacol (0.34 g, 2.91 mmol), magnesium sulfate (1.5 g, 12.46 mmol) and acetic acid (0.1 mL) was stirred at ambient temperature for 18 h. The mixture was filtered and the filtrate was washed with brine, dried (Na.sub.2SO.sub.4), concentrated to provide the title compound. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.40 (s, 12H), 7.49 (dd, J=7.71, 4.78 Hz, 1H), 8.18 (dd, J=7.81, 1.76 Hz, 1H), 8.75 (dd, J=4.78, 1.86 Hz, 1H).

Step B

3-(3-Amino-4-(tert-butylthio)phenyl)picolinonitrile

(173) ##STR00085##

(174) A mixture of 5-bromo-2-(tert-butylthio)aniline (200 mg, 0.769 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinonitrile (230 mg, 0.999 mmol), cesium fluoride (350 mg, 2.306 mmol), copper(I) iodide (29.3 mg, 0.154 mmol), tetrakis(triphenylphosphine) palladium (0) (178 mg, 0.154 mmol) and 1,4-Dioxane (6 mL) was degassed with a stream of nitrogen for 5 min then placed in an oil bath at 100 C. under nitrogen atmosphere for 2.5 h. The mixture was diluted with water and extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified on silica gel (EtOAc/hexanes 0-20%) to provide the title compound (108.5 mg, 0.371 mmol, 48.3% yield).). .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.36 (s, 9H), 6.83 (dd, J=7.91, 1.86 Hz, 1H), 6.93 (d, J=1.56 Hz, 1H), 7.50 (d, J=8.01 Hz, 1H), 7.56 (dd, J=8.01, 4.69 Hz, 1H), 7.86 (dd, J=8.01, 1.56 Hz, 1H), 8.69 (dd, J=4.69, 1.37 Hz, 1H).

Step C

N-(2-(tert-Butylthio)-5-(2-cyanopyridin-3-yl)phenyl)-2-(p-tolyl)acetamide

(175) ##STR00086##

(176) To a solution of 3-(3-amino-4-(tert-butylthio)phenyl)picolinonitrile (76 mg, 0.228 mmol) in Ethyl acetate (2.5 mL) was added 2-(p-tolyl)acetic acid (41.1 mg, 0.274 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (174 mg, 0.274 mmol) (50%/EtOAc) followed by dropwise addition of DIEA (0.119 mL, 0.684 mmol) and the mixture was stirred at ambient temperature for 3 h. More 2-(p-tolyl)acetic acid (20 mg), 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (87 mg) and DIEA (0.060 mL) was added. After 18 h the mixture was washed with water and the organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified by HPLC (RP C18 MeCN/water 10-100%, 0.1% formic acid) to provide the title compound (43.8 mg, 0.103 mmol, 45.3% yield). LCMS (M+1).sup.+: m/z=416.3. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.06 (s, 9H), 2.38 (s, 3H), 3.77 (s, 2H), 7.20-7.28 (m, 4H), 7.52-7.64 (m, 2H), 7.93 (d, J=8.01 Hz, 1H), 8.71 (d, J=4.49 Hz, 1H), 8.79 (s, 1H), 8.94 (s, 1H).

Step D

N-(5-(2-(2H-Tetrazol-5-yl)pyridin-3-yl)-2-(tert-butylthio)phenyl)-2-(p-tolyl)acetamide

(177) ##STR00087##

(178) To a solution of N-(2-(tert-butylthio)-5-(2-cyanopyridin-3-yl)phenyl)-2-(p-tolyl)acetamide (42 mg, 0.099 mmol) in 1-propanol (2.5 mL) was added sodium azide (12.88 mg, 0.198 mmol) and zinc chloride (0.594 mL, 0.297 mmol) (0.5 M/THF) and the mixture was heated at 95 C. under nitrogen atmosphere for 24 h. The mixture was filtered and purified by HPLC (RP C18 MeCN/water 10-100%, 0.1% formic acid) to provide the title compound (6.5 mg, 0.013 mmol, 13.59% yield). LCMS (M+1).sup.+: m/z=459.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.03 (s, 9H), 2.31 (s, 3H), 3.72 (s, 2H), 6.92 (d, J=6.84 Hz, 1H), 7.19-7.25 (m, 2H), 7.29 (d, J=8.01 Hz, 2H), 7.42 (d, J=7.81 Hz, 1H), 7.71 (dd, J=7.71, 4.78 Hz, 1H), 8.00 (d, J=7.62 Hz, 1H), 8.23 (s, 1H), 8.81 (d, J=3.91 Hz, 1H), 9.00 (s, 1H).

(179) ##STR00088##

Example 64: N-(5-(2-(1H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(p-tolyl)acetamide

(180) ##STR00089##

Step A

5-Bromo-2-(tert-butylthio)-3-nitropyridine

(181) ##STR00090##

(182) This compound was prepared from 5-bromo-2-chloro-3-nitropyridine following the procedure described in Step A (Scheme I). .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.58 (s, 9H), 8.73 (d, J=1.83 Hz, 1H), 8.97 (d, J=1.65 Hz, 1H).

Step B

5-Bromo-2-(tert-butylthio)pyridin-3-amine

(183) ##STR00091##

(184) This compound was prepared from 5-bromo-2-(tert-butylthio)-3-nitropyridine following the procedure described in Step B (Scheme I). .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.39 (s, 9H), 5.60 (br. s., 2H), 7.20 (d, J=1.83 Hz, 1H), 7.84 (d, J=1.83 Hz, 1H).

Step C

N-(5-Bromo-2-(tert-butylthio)pyridin-3-yl)-2-(p-tolyl)acetamide

(185) ##STR00092##

(186) This compound was prepared from 5-bromo-2-(tert-butylthio)pyridin-3-amine following the procedure described in Step C (Scheme I). .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.36 (s, 9H), 2.29 (s, 3H), 3.71 (s, 2H), 7.13-7.21 (m, 2H), 7.26 (d, J=7.51 Hz, 2H), 8.31 (br. s., 1H), 8.45 (d, J=1.47 Hz, 1H), 9.35 (s, 1H).

Step D

N-(5-(2-(1H-Tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(p-tolyl)acetamide

(187) ##STR00093##

(188) This compound was prepared from N-(5-bromo-2-(tert-butylthio)pyridin-3-yl)-2-(p-tolyl)acetamide following the procedure described in Step D (Scheme I). .sup.1H NMR (400 MHz, METHANOL-d.sub.4): ppm 1.21 (s, 9H), 2.36 (s, 3H), 3.74 (s, 2H), 7.28 (q, J=7.69 Hz, 4H), 7.60-7.69 (m, 2H), 7.75 (dd, J=18.86, 7.51 Hz, 2H), 8.00 (d, J=1.65 Hz, 1H), 8.39 (s, 1H).

(189) The following examples were synthesized following the procedures described in Scheme X.

Example 65: N-(5-(2-(1H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(4-(trifluoromethyl)phenyl)acetamide

(190) ##STR00094##

(191) .sup.1H NMR (400 MHz, METHANOL-d.sub.4): ppm 1.28 (s, 9H), 3.88 (br. s., 2H), 7.57-7.66 (m, 4H), 7.70 (d, J=7.14 Hz, 4H), 7.76 (d, J=7.33 Hz, 1H), 8.02 (br. s., 1H), 8.17 (br. s., 1H).

Example 66: N-(5-(2-(1H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(2,4-dimethylphenyl)acetamide

(192) ##STR00095##

(193) .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.31 (s, 9H), 2.22 (s, 3H), 2.26 (s, 3H), 3.70 (s, 2H), 6.96-7.09 (m, 1H), 7.13-7.28 (m, 1H), 7.28-7.46 (m, 2H), 7.46-7.71 (m, 4H), 7.76 (d, J=7.51 Hz, 1H), 8.01 (br. s., 1H), 9.02 (s, 1H).

Example 67: N-(5-(2-(1H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(2-(trifluoromethyl)phenyl)acetamide

(194) ##STR00096##

(195) .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.43 (br. s., 9H), 3.96 (br. s., 2H), 7.49-7.82 (m, 9H), 8.05 (br. s., 1H), 9.34 (br. s., 1H).

Example 68: N-(5-(2-(1H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(4-cyclopropylphenyl)acetamide

(196) ##STR00097##

(197) .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.54-0.73 (m, 2H), 0.86-1.00 (m, 2H), 1.35 (s, 9H), 1.78-2.00 (m, 1H), 3.66 (br. s., 2H), 7.06 (d, J=7.87 Hz, 2H), 7.22 (d, J=7.51 Hz, 2H), 7.56-7.67 (m, 2H), 7.67-7.80 (m, 2H), 7.89 (br. s., 1H), 8.02 (s, 1H), 9.19 (s, 1H).

(198) ##STR00098##

Example 69: N-(5-(2-(2H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(4-isopropylphenyl)acetamide

(199) ##STR00099##

Step A

5-(2-(2H-Tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-amine

(200) ##STR00100##

(201) This compound was prepared from 5-bromo-2-(tert-butylthio)pyridin-3-amine following the procedure described in Step A (Scheme V). LCMS (M+1).sup.+: m/z=327.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.40 (s, 9H), 5.33 (s, 2H), 6.75 (d, J=1.95 Hz, 1H), 7.45 (d, J=2.15 Hz, 1H), 7.59 (d, J=6.64 Hz, 3H), 7.64-7.77 (m, 1H), 8.00-8.10 (m, 1H).

Step B

N-(5-(2-(2H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-(4-isopropylphenyl)acetamide

(202) ##STR00101##

(203) This compound was prepared from 5-(2-(2H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-amine following the procedure described in Step B (Scheme V). LCMS (M+1).sup.+: m/z=487.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.19 (d, J=6.84 Hz, 6H), 1.35 (s, 9H), 2.87 (quin, J=6.88 Hz, 1H), 3.68 (s, 2H), 7.25 (q, J=8.14 Hz, 4H), 7.56-7.68 (m, 2H), 7.68-7.80 (m, 2H), 7.89 (s, 1H), 8.02 (d, J=1.76 Hz, 1H), 9.24 (s, 1H).

Example 70: N-(5-(2-(2H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-yl)-2-mesitylacetamide

(204) ##STR00102##

(205) This compound was prepared from 5-(2-(2H-tetrazol-5-yl)phenyl)-2-(tert-butylthio)pyridin-3-amine following the procedure described in Step B (Scheme V). LCMS (M+1).sup.+: m/z 487.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.27 (s, 9H), 2.21-2.28 (m, 9H), 3.72 (s, 2H), 6.92 (s, 2H), 7.58-7.68 (m, 2H), 7.69-7.81 (m, 2H), 8.01 (s, 1H), 8.07 (s, 1H), 8.94 (s, 1H).

(206) ##STR00103##

Example 71: N-(4-(tert-butylthio)-2-(1H-1,2,3-triazol-4-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(207) ##STR00104##

Step A

4-(tert-Butylthio)-2-ethynyl-[1,1-biphenyl]-3-amine

(208) ##STR00105##

(209) This compound was prepared from 5-bromo-2-(tert-butylthio)aniline and(2-ethynylphenyl)boronic acid following procedure described in Step A (Scheme V). LCMS (M+1).sup.+: m/z 282.2. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.34-1.41 (m, 9H), 3.08 (s, 1H), 6.91 (dd, J=7.91, 1.86 Hz, 1H), 6.98 (d, J=1.76 Hz, 1H), 7.28 (d, J=13.47 Hz, 1H), 7.35-7.48 (m, 3H), 7.61 (d, J=7.81 Hz, 1H).

Step B

N-(4-(tert-butylthio)-2-ethynyl-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(210) ##STR00106##

(211) This compound was prepared from 4-(tert-butylthio)-2-ethynyl-[1,1-biphenyl]-3-amine following the procedure described in Step B (Scheme V). LCMS (M+1).sup.+: m/z 414.3. .sup.1H NMR (400 MHz, CHLOROFORM-d): ppm 1.06 (s, 9H), 2.38 (s, 3H), 3.07 (s, 1H), 3.76 (s, 2H), 7.22-7.35 (m, 5H), 7.37-7.50 (m, 4H), 7.61 (d, J=7.69 Hz, 1H), 8.79 (s, 1H), 8.87 (br. s., 1H).

Step C

N-(4-(tert-butylthio)-2-(1H-1,2,3-triazol-4-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(212) ##STR00107##

(213) To a solution of N-(4-(tert-butylthio)-2-ethynyl-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide (22 mg, 0.045 mmol) in DMF (0.8 mL)/methanol (0.200 mL) was added copper(I) iodide (0.861 mg, 4.52 mol) and TMS-N.sub.3 (0.012 mL, 0.090 mmol) and the mixture was stirred at 100 C. for 5 h. More copper (I) iodide (5 mg) and TMS-N.sub.3 (0.40 mL) was added and the mixture was stirred for 30 min at 100 C. Water was added and the mixture was extracted with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), concentrated and purified by HPLC (RP C18, MeCN/water 10-100%, 0.1% formic acid) to provide the title compound (3 mg, 6.50 mol, 14.39% yield). LCMS (M+1).sup.+: m/z 457.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.03 (s, 9H), 2.31 (s, 3H), 3.72 (s, 2H), 6.80-6.95 (m, 1H), 7.18-7.25 (m, 2H), 7.26-7.32 (m, 2H), 7.33-7.60 (m, 4H), 7.77 (br. s., 1H), 8.21 (br. s., 1H), 9.01 (br. s., 1H).

(214) ##STR00108##

Example 72: N-(4-(tert-butylthio)-2-(1H-1,2,4-triazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(215) ##STR00109##

Step A

4-(tert-Butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxamide

(216) ##STR00110##

(217) This compound was prepared from N-(5-bromo-2-(tert-butylthio)phenyl)-2-(p-tolyl)acetamide and (2-carbamoylphenyl)boronic acid following the procedure described in Step D (Scheme I). LCMS (M+1).sup.+: m/z 433.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.04 (s, 9H), 2.31 (s, 3H), 3.75 (s, 2H), 7.11 (dd, J=7.91, 1.86 Hz, 1H), 7.20-7.26 (m, 2H), 7.28-7.38 (m, 4H), 7.39-7.55 (m, 4H), 7.71 (s, 1H), 8.41 (d, J=1.76 Hz, 1H), 9.01 (s, 1H).

Step B

N-(4-(tert-butylthio)-2-(1H-1,2,4-triazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(218) ##STR00111##

(219) A mixture of 4-(tert-butylthio)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxamide (118 mg, 0.273 mmol) and DMF-DMA (2 mL, 14.94 mmol) was heated at 80 C. for 1 h and concentrated to provide (E)-4-(tert-butylthio)-N-((dimethylamino)methylene)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxamide (139.5 mg, 0.286 mmol, 105% yield) used as is in the next step. To a portion of this product (69 mg, 0.141 mmol) in Acetic Acid (1 mL) was added hydrazine (6.66 l, 0.212 mmol) (hydrazine monohydrate, 0.01 mL) and the mixture was stirred at 110 C. for 1.5 h. The mixture was concentrated and purified by HPLC (RP C18, MeCN/water 10-100%, 0.1% formic acid) to provide the title compound (15.3 mg, 0.034 mmol, 23.68% yield). LCMS (M+1).sup.+: m/z 457.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 0.94-1.07 (m, 9H), 2.31 (s, 3H), 3.71 (s, 2H), 6.76 (d, J=7.62 Hz, 1H), 7.19-7.25 (m, 2H), 7.26-7.34 (m, 3H), 7.38 (d, J=6.64 Hz, 1H), 7.43-7.66 (m, 2H), 7.70-7.89 (m, 1H), 8.14-8.29 (m, 1H), 8.43 (s, 1H), 8.86-9.05 (m, 1H).

Example 73: N-(4-(tert-butylthio)-2-(1,2,4-oxadiazol-5-yl)-[1,1-biphenyl]-3-yl)-2-(p-tolyl)acetamide

(220) ##STR00112##

(221) To a solution of (E)-4-(tert-butylthio)-N-((dimethylamino)methylene)-3-(2-(p-tolyl)acetamido)-[1,1-biphenyl]-2-carboxamide (69 mg, 0.141 mmol) prepared as described above in 1,4-dioxane (1.000 mL) was added a mixture made by adding hydroxylamine hydrochloride (11.80 mg, 0.170 mmol) to sodium hydroxide (0.031 mL, 0.156 mmol) (5M/water) and acetic acid (0.3 mL) and the mixture was stirred at 90 C. for 4.5 h. The mixture was concentrated, diluted with DMF and purified by HPLC (RP C18, MeCN/water 20-100%, 0.1% formic acid) to provide the title compound (15 mg, 0.033 mmol, 23.17% yield).). LCMS (M+1).sup.+: m/z 458.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): ppm 1.03 (s, 9H), 2.31 (s, 3H), 3.72 (s, 2H), 6.91 (dd, J=7.81, 1.95 Hz, 1H), 7.19-7.25 (m, 2H), 7.26-7.32 (m, 2H), 7.44 (d, J=7.81 Hz, 1H), 7.57 (d, J=7.81 Hz, 1H), 7.63-7.70 (m, 1H), 7.73-7.82 (m, 1H), 8.00 (d, J=7.03 Hz, 1H), 8.22 (d, J=1.56 Hz, 1H), 8.95-9.04 (m, 2H).

(222) ##STR00113##

Example 74: N-(4-(tert-Butylthio)-2-(1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-5-chloropyridin-2-amine

(223) ##STR00114##

Step A

N-(5-bromo-2-(tert-butylthio)phenyl)-5-chloropyridin-2-amine

(224) ##STR00115##

(225) A mixture of 5-bromo-2-(tert-butylthio)aniline (300 mg, 1.2 mmol), 5-chloro-2-fluoropyridine (157 mg, 1.2 mmol), Cs.sub.2CO.sub.3 (782 mg, 2.4 mmol) in NMP (5 mL) was purged with N.sub.2 before heated to 140 C. for 8 h. The reaction mixture was then cooled down to room temperature. Water was added and the mixture was partitioned between EtOAc (50 mL) and water (15 mL). The layers were separated and the organic layer was washed with brine (20 mL), dried with Na.sub.2SO.sub.4, and concentrated. The crude product was purified by flash chromatography (silica gel, 5-20% EtOAc in petroleum ether) to give the title compound (100 mg, 24%) as a yellow solid. LCMS (M+H).sup.+: m/z=371.43.

Step B

N-(4-(tert-butylthio)-2-(1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-5-chloropyridin-2-amine

(226) ##STR00116##

(227) A mixture of N-(5-bromo-2-(tert-butylthio)phenyl)-5-chloropyridin-2-amine (100 mg, 0.27 mmol), (2-(1H-tetrazol-5-yl)phenyl)boronic acid (51 mg, 0.27 mmol), Pd(PPh.sub.3).sub.4 (32 mg, 0.027 mmol), K.sub.2CO.sub.3 (112 mg, 0.81 mmol) in DMF/H.sub.2O (2.5 mL/0.5 mL) was purged with N.sub.2 before heated to 95 C. overnight. The reaction mixture was then cooled down to room temperature and filtered off the solid. The filtrate was concentrated and partitioned between EtOAc (15 mL) and water (10 mL). The layers were separated and the organic layer was washed with brine (15 mL), dried with Na.sub.2SO.sub.4, and concentrated. The residue was purified by preparative HPLC (RP C18 column, 40-100% acetonitrile in water, 0.1% formic acid) to give the titled compound (30 mg, 26%) as a white solid. LCMS (M+H).sup.+: m/z=437.22. .sup.1H NMR (400 MHz, CDCl.sub.3): 15.35 (s, 1H), 8.22-8.14 (m, 2H), 8.07 (d, J=1.9 Hz, 1H), 7.93 (s, 1H), 7.59 (ddd, J=14.3, 7.4, 4.3 Hz, 4H), 7.47-7.42 (m, 1H), 6.85 (dd, J=7.8, 1.9 Hz, 1H), 6.81 (d, J=8.8 Hz, 1H), 1.28 (s, 9H).

(228) ##STR00117##

Example 75: 6-((4-(tert-Butylthio)-2-(1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)amino)nicotinonitrile

(229) ##STR00118##

Step A

6-((5-bromo-2-(tert-butylthio)phenyl)amino)nicotinonitrile

(230) ##STR00119##

(231) A mixture of 5-bromo-2-(tert-butylthio)aniline (300 mg, 1.2 mmol) and 6-fluoronicotinonitrile (440 mg, 3.6 mmol) was place in a microwave vial and stirred at 180 C. under neat condition for 5 h before cooled down to room temperature. EtOAc and water were added and the organic layer was washed with brine, dried over Na.sub.2SO.sub.4, concentrated under vacuum and purified on silica gel (5-60% EtOAc in petroleum ether) to get the title compound (260 mg, 62% yield) as a yellow solid. LCMS (M+H).sup.+: m/z=362.09.

Step B

6-((4-(tert-butylthio)-2-(1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)amino)nicotinonitrile

(232) ##STR00120##

(233) A mixture of 6-((5-bromo-2-(tert-butylthio)phenyl)amino)nicotinonitrile (130 mg, 0.36 mmol), (2-(1H-tetrazol-5-yl)phenyl)boronic acid (68 mg, 0.36 mmol), Pd(PPh.sub.3).sub.4 (42 mg, 0.036 mmol), K.sub.2CO.sub.3 (149 mg, 1.08 mmol) in DMF/H.sub.2O (2.5 mL/0.5 mL) was purged with N.sub.2 before heated to 95 C. overnight. The reaction mixture was then cooled down to room temperature and filtered off the solid. The filtrate was concentrated and partitioned between EtOAc (15 mL) and water (10 mL). The layers were separated and the organic layer was washed with brine (15 mL), dried with Na.sub.2SO.sub.4, and concentrated. The residue was purified by preparative HPLC (RP C18 column, 40-100% acetonitrile in water, 0.1% formic acid) to give the titled compound (39 mg, 25%) as a white solid. LCMS (M+H).sup.+: m/z=428.35. .sup.1H NMR (400 MHz, CDCl.sub.3): 8.53 (d, J=2.0 Hz, 1H), 8.27-8.19 (m, 2H), 8.15 (d, J=1.9 Hz, 1H), 7.80 (dd, J=8.7, 2.2 Hz, 1H), 7.66-7.57 (m, 3H), 7.48-7.44 (m, 1H), 6.96 (dd, J=7.9, 1.9 Hz, 1H), 6.86 (d, J=8.7 Hz, 1H), 1.29 (s, 9H).

Example 76: N-(4-(tert-butylthio)-2-(1H-tetrazol-5-yl)-[1,1-biphenyl]-3-yl)-5-(trifluoromethyl)pyridin-2-amine

(234) ##STR00121##

(235) This compound was prepared following Steps A and B (Scheme XIV) to provide the title compound. LCMS (M+H).sup.+: m/z=471.26. .sup.1H NMR (400 MHz, DMSO): 8.80 (s, 1H), 8.42 (s, 1H), 7.95 (d, J=1.7 Hz, 1H), 7.86 (dd, J=8.9, 2.5 Hz, 1H), 7.73-7.67 (m, 2H), 7.65-7.58 (m, 2H), 7.50 (d, J=7.9 Hz, 1H), 7.13 (d, J=8.9 Hz, 1H), 6.90 (dd, J=7.9, 2.0 Hz, 1H).

Compound Data

(236) Human indoleamine 2,3-dioxgenase (IDO) cellular data is presented below. Brief descriptions of the cellular assays are provided following the table.

(237) TABLE-US-00001 Example HeLa pIC.sub.50 PBMC pIC.sub.50 1 7.8 7.9 4 6.5 6.1 5 <5 6 5.2 7 5.3 8 5.2 9 <5 10 6.6 6.9 11 6 12 5.3 13 6.4 7.1 14 5.5 5.8 15 5.7 5.7 16 6.6 17 5.6 18 5.4 19 7.8 8 20 7.6 7.7 21 7.7 22 7.5 23 7.2 24 6.4 25 6.4 26 6.1 27 8.4 8.5 29 5.3 30 <5 31 <5 32 <5 33 5.1 34 7.1 7.6 35 <5.5 5.8 36 5.6 5.9 37 6.1 6.6 38 7 39 8.6 8.8 40 7.5 7.7 42 6.9 43 7.3 7.9 44 5.5 5.4 45 5.6 46 5.2 5.3 47 6.3 6.3 48 5.5 49 6 50 6.9 51 7.8 7.2 52 7.6 6.8 55 7.5 7.3 56 7.5 7.7 57 7.6 58 7.4 59 6.4 60 7.2 61 6.2 62 6.3 64 7.2 7.3 65 7.4 7.6 66 8 8.1 67 7 7.9

(238) HeLa IDOi Assay:

(239) Compounds of the present invention were tested via high-throughput cellular assays utilizing detection of kynurenine via mass spectrometry and cytotoxicity as end-points. For the mass spectrometry and cytotoxicity assays, human epithelial HeLa cells (CCL-2; ATCC, Manassas, Va.) were stimulated with human interferon- (IFN-) (Sigma-Aldrich Corporation, St. Louis, Mo.) to induce the expression of indoleamine 2,3-dioxygenase (IDO1). Compounds with IDO1 inhibitory properties decreased the amount of kynurenine produced by the cells via the tryptophan catabolic pathway. Cellular toxicity due to the effect of compound treatment was measured using CellTiter-Glo reagent (CTG) (Promega Corporation, Madison, Wis.), which is based on luminescent detection of ATP, an indicator of metabolically active cells.

(240) In preparation for the assays, test compounds were serially diluted 3-fold in DMSO from a typical top concentration of 5 mM and plated at 0.5 L in 384-well, polystyrene, clear bottom, tissue culture treated plates with lids (Greiner Bio-One, Kremsmnster, Austria) to generate 11-point dose response curves. Low control wells (0% kynurenine or 100% cytotoxicity) contained either 0.5 L of DMSO in the presence of unstimulated (IFN-) HeLa cells for the mass spectrometry assay or 0.5 L of DMSO in the absence of cells for the cytotoxicity assay, and high control wells (100% kynurenine or 0% cytotoxicity) contained 0.5 L of DMSO in the presence of stimulated (+IFN-) HeLa cells for both the mass spectrometry and cytotoxicity assays.

(241) Frozen stocks of HeLa cells were washed and recovered in DMEM high glucose medium with HEPES (Thermo Fisher Scientific, Inc., Waltham, Mass.) supplemented with 10% v/v certified fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc., Waltham, Mass.), and 1 penicillin-streptomycin antibiotic solution (Thermo Fisher Scientific, Inc., Waltham, Mass.). The cells were diluted to 100,000 cells/mL in the supplemented DMEM medium. 50 L of either the cell suspension, for the mass spectrometry assay, or medium alone, for the cytotoxicity assay, were added to the low control wells, on the previously prepared 384-well compound plates, resulting in 5,000 cells/well or 0 cells/well respectively. IFN- was added to the remaining cell suspension at a final concentration of 10 nM, and 50 L of the stimulated cells were added to all remaining wells on the 384-well compound plates. The plates, with lids, were then placed in a 37 C., 5% CO.sub.2 humidified incubator for 2 days.

(242) Following incubation, the 384-well plates were removed from the incubator and allowed to equilibrate to room temperature for 30 minutes. For the cytotoxicity assay, CellTiter-Glo was prepared according to the manufacturer's instructions, and 10 L were added to each plate well. After a twenty minute incubation at room temperature, luminescence was read on an EnVision Multilabel Reader (PerkinElmer Inc., Waltham, Mass.). For the mass spectrometry assay, 10 L of supernatant from each well of the compound-treated plates were added to 40 L of acetonitrile, containing 10 M of an internal standard for normalization, in 384-well, polypropylene, V-bottom plates (Greiner Bio-One, Kremsmnster, Austria) to extract the organic analytes. Following centrifugation at 2000 rpm for 10 minutes, 10 L from each well of the acetonitrile extraction plates were added to 90 L of sterile, distilled H.sub.2O in 384-well, polypropylene, V-bottom plates for analysis of kynurenine and the internal standard on the RapidFire 300 (Agilent Technologies, Santa Clara, Calif.) and 4000 QTRAP MS (SCIEX, Framingham, Mass.). MS data were integrated using Agilent Technologies' RapidFire Integrator software, and data were normalized for analysis as a ratio of kynurenine to the internal standard.

(243) The data for dose responses in the mass spectrometry assay were plotted as IDO1 inhibition versus compound concentration following normalization using the formula 100(100*((UC2)/(C1C2))), where U was the unknown value, C1 was the average of the high (100% kynurenine; 0% inhibition) control wells and C2 was the average of the low (0% kynurenine; 100% inhibition) control wells. The data for dose responses in the cytotoxicity assay were plotted as % cytotoxicity versus compound concentration following normalization using the formula 100(100*((UC2)/(C1C2))), where U was the unknown value, C1 was the average of the high (0% cytotoxicity) control wells and C2 was the average of the low (100% cytotoxicity) control wells.

(244) Curve fitting was performed with the equation y=A+((BA)/(1+(10.sup.x/10.sup.C).sup.D)), where A was the minimum response, B was the maximum response, C was the log(XC.sub.50) and D was the Hill slope. The results for each test compound were recorded as pIC50 values for the mass spectrometry assay and as pCC50 values for the cytoxicity assay (C in the above equation).

(245) PBMC IDOi Assay:

(246) Compounds of the present invention were tested via high-throughput cellular assays utilizing detection of kynurenine via mass spectrometry and cytotoxicity as end-points. For the mass spectrometry and cytotoxicity assays, human peripheral blood mononuclear cells (PBMC) (PB003F; AllCells, Alameda, Calif.) were stimulated with human interferon- (IFN-) (Sigma-Aldrich Corporation, St. Louis, Mo.) and lipopolysaccharide from Salmonella minnesota (LPS) (Invivogen, San Diego, Calif.) to induce the expression of indoleamine 2,3-dioxygenase (IDO1). Compounds with IDO1 inhibitory properties decreased the amount of kynurenine produced by the cells via the tryptophan catabolic pathway. Cellular toxicity due to the effect of compound treatment was measured using CellTiter-Glo reagent (CTG) (Promega Corporation, Madison, Wis.), which is based on luminescent detection of ATP, an indicator of metabolically active cells.

(247) In preparation for the assays, test compounds were serially diluted 3-fold in DMSO from a typical top concentration of 5 mM and plated at 0.5 L in 384-well, polystyrene, clear bottom, tissue culture treated plates with lids (Greiner Bio-One, Kremsmnster, Austria) to generate 11-point dose response curves. Low control wells (0% kynurenine or 100% cytotoxicity) contained either 0.5 L of DMSO in the presence of unstimulated (IFN-/LPS) PBMCs for the mass spectrometry assay or 0.5 L of DMSO in the absence of cells for the cytotoxicity assay, and high control wells (100% kynurenine or 0% cytotoxicity) contained 0.5 L of DMSO in the presence of stimulated (+IFN-/+LPS) PBMCs for both the mass spectrometry and cytotoxicity assays.

(248) Frozen stocks of PBMCs were washed and recovered in RPMI 1640 medium (Thermo Fisher Scientific, Inc., Waltham, Mass.) supplemented with 10% v/v heat-inactivated fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc., Waltham, Mass.), and 1 penicillin-streptomycin antibiotic solution (Thermo Fisher Scientific, Inc., Waltham, Mass.). The cells were diluted to 1,000,000 cells/mL in the supplemented RPMI 1640 medium. 50 L of either the cell suspension, for the mass spectrometry assay, or medium alone, for the cytotoxicity assay, were added to the low control wells, on the previously prepared 384-well compound plates, resulting in 50,000 cells/well or 0 cells/well respectively. IFN- and LPS were added to the remaining cell suspension at final concentrations of 100 ng/ml and 50 ng/ml respectively, and 50 L of the stimulated cells were added to all remaining wells on the 384-well compound plates. The plates, with lids, were then placed in a 37 C., 5% CO.sub.2 humidified incubator for 2 days.

(249) Following incubation, the 384-well plates were removed from the incubator and allowed to equilibrate to room temperature for 30 minutes. For the cytotoxicity assay, CellTiter-Glo was prepared according to the manufacturer's instructions, and 40 L were added to each plate well. After a twenty minute incubation at room temperature, luminescence was read on an EnVision Multilabel Reader (PerkinElmer Inc., Waltham, Mass.). For the mass spectrometry assay, 10 L of supernatant from each well of the compound-treated plates were added to 40 L of acetonitrile, containing 10 M of an internal standard for normalization, in 384-well, polypropylene, V-bottom plates (Greiner Bio-One, Kremsmnster, Austria) to extract the organic analytes. Following centrifugation at 2000 rpm for 10 minutes, 10 L from each well of the acetonitrile extraction plates were added to 90 L of sterile, distilled H.sub.2O in 384-well, polypropylene, V-bottom plates for analysis of kynurenine and the internal standard on the RapidFire 300 (Agilent Technologies, Santa Clara, Calif.) and 4000 QTRAP MS (SCIEX, Framingham, Mass.). MS data were integrated using Agilent Technologies' RapidFire Integrator software, and data were normalized for analysis as a ratio of kynurenine to the internal standard.

(250) The data for dose responses in the mass spectrometry assay were plotted as % IDO1 inhibition versus compound concentration following normalization using the formula 100(100*((UC2)/(C1C2))), where U was the unknown value, C1 was the average of the high (100% kynurenine; 0% inhibition) control wells and C2 was the average of the low (0% kynurenine; 100% inhibition) control wells. The data for dose responses in the cytotoxicity assay were plotted as % cytotoxicity versus compound concentration following normalization using the formula 100(100*((UC2)/(C1C2))), where U was the unknown value, C1 was the average of the high (0% cytotoxicity) control wells and C2 was the average of the low (100% cytotoxicity) control wells.

(251) Curve fitting was performed with the equation y=A+((BA)/(1+(10.sup.x/10.sup.C).sup.D)), where A was the minimum response, B was the maximum response, C was the log(XC.sub.50) and D was the Hill slope. The results for each test compound were recorded as pIC50 values for the mass spectrometry assay and as pCC50 values for the cytoxicity assay (C in the above equation).