BRIARANE COMPOUNDS AND USE THEIR USE

Abstract

Disclosed are compounds compound of Formula (I):

##STR00001## Each variable is defined herein. Also included are pharmaceutical compositions containing such a compound and a method of treating an inflammatory condition.

Claims

1. A compound of Formula (I): ##STR00109## in which R.sub.1 is C.sub.1-C.sub.6 alkyl; R.sub.2 is C.sub.1-C.sub.6 alkyl, formyl, carboxyl, CH?NOR.sub.a, CH?NR.sub.a, or C(O)NHOR.sub.a, in which R.sub.a is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl, or C.sub.1-C.sub.6 heterocycloalkyl; R.sub.3 is H, hydroxy, or combined with R.sub.4, and R.sub.4 is H or C.sub.1-C.sub.6 alkyl; when R.sub.3 and R.sub.4 are combined, they together with the two carbon atoms attached thereto form an epoxide ring; R.sub.5 is N.sub.3, methyl, or combined with X.sub.2, and when R.sub.5 and X.sub.2 are combined, they are N and, together with the carbon atoms attached thereto, form a heterocycloalkyl ring; R.sub.6 is H, halo, oxo, hydroxyl, ?NR.sub.b, ?NNHC(O)R.sub.c, OC(O)R.sub.d, or OS(O).sub.2R.sub.e, in which R.sub.b is hydroxyl, C.sub.1-C.sub.6 alkoxy, or C.sub.1-C.sub.10 heterocycloalkyl, R.sub.c is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6 cycloalkyl, 5 or 6-membered heterocycloalkyl, phenyl, or 5- or 6-membered heteroaryl, R.sub.d is C.sub.3-C.sub.6 cycloalkyl, C.sub.2-C.sub.10 alkenyl, C.sub.1-C.sub.10 aminoalkyl, C.sub.3-C.sub.10 alkynylalkyl, C.sub.1-C.sub.10 heterocyclylalkyl, or phenyl, and R.sub.e is amino or C.sub.1-C.sub.6 alkyl; R.sub.7 is H or acetyloxy; X.sub.1 is hydroxyl or oxo; X.sub.2 is O or N, when X.sub.2 is N, it is combined with R.sub.5; each of is a single bond or a double bond; alkyl is unsubstituted or substituted with one or more groups selected from hydroxyl, amino, CN, aryl, heteroaryl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 carboxylate, and C.sub.1-C.sub.6 heterocycloalkyl; each of amino, aminoalkyl, alkenyl, alkoxy, cycloalkyl, alkynylalkyl, heterocyclylalkyl, heterocycloalkyl, heteroaryl, and phenyl, is unsubstituted or substituted with one or more groups selected from hydroxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 carboxylate, C.sub.1-C.sub.6 heterocycloalkyl, aryl, heteroaryl, and aryl-amino-alkyl-aminocarbonyl.

2. The compound of claim 1, wherein R.sub.2 is formyl, carboxyl, CH?NOR.sub.a, CH?NR.sub.a, or C(O)NHOR.sub.a; and R.sub.6 is OH.

3. The compound of claim 1, wherein each of X.sub.1 and R.sub.6 is OH.

4. The compound of claim 1, wherein R.sub.6 is oxo, ?NR.sub.b, ?NNHC(O)R.sub.c, OC(O)R.sub.d, or OS(O).sub.2R.sub.e.

5. The compound of claim 1, wherein R.sub.1 is methyl, ethyl, n-propyl, or iso-propyl.

6. The compound of claim 1, wherein each of R.sub.3 and R.sub.4, independently, is H or OH, or R.sub.3 and R.sub.4 are combined and, together with the carbon atoms they bond to, form an epoxide ring.

7. The compound of claim 1, wherein R.sub.5 is H, OH, N.sub.3, or methyl.

8. The compound of claim 1, wherein R.sub.7 is acetyloxy.

9. The compound of claim 1, wherein the compound is one of Formulas (I-A), (I-B), (I-C), (I-D), (I-E), (I-F), and (I-G): ##STR00110## ##STR00111##

10. The compound of claim 9, wherein the compound is of Formula (I-A), and R.sub.b is hydroxyl, C.sub.1-C.sub.6 alkoxy, 5- or 6-membered heterocycloalkyl, or NHC(O)R.sub.c, R.sub.c being C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6 cycloalkyl, phenyl, or heteroaryl.

11. The compound of claim 10, wherein R.sub.1 is propyl, R.sub.7 is acetyloxy, and R.sub.b is hydroxyl, methoxy, CH?CCH.sub.2O, PhC(O)NH, CH.sub.3C(O)NH, cyclopropanecarbox-amido, N-methylpiperazin-1-yl, or morpholino.

12. The compound of claim 9, wherein the compound is of Formula (I-B) or (I-C), wherein R.sub.a is H, methyl, ethyl, prop-2-yn-1-yl, prop-2-en-1-yl, morpholino, benzyl, CH.sub.2CN, CH.sub.2CH.sub.2N(CH.sub.3).sub.2, or ##STR00112##

13. The compound of claim 9, wherein R.sub.1 is propyl and R.sub.7 is acetyloxy.

14. The compound of claim 9, wherein the compound is of Formula (I-B).

15. The compound of claim 9, wherein the compound if of Formula (I-C).

16. The compound of claim 9, wherein the compound is of Formula (I-D), (I-E), (I-F), or (I-G), wherein each of R.sub.d and R.sub.e, independently, is methyl, but-3-yn-1-yl, t-butyl-OC(O)NHCH.sub.2, CH.sub.3).sub.2NCH.sub.2, NH.sub.2, CH.sub.2NH.sub.2, CH.sub.2CH.sub.2NH.sub.2, CH.sub.2CH.sub.2N(CH.sub.3).sub.2, CH.sub.2NHC(O)CH.sub.3, CH.sub.2(CH.sub.2).sub.4NH.sub.2, CH.sub.2(CH.sub.2).sub.4N(CH.sub.3).sub.2, ##STR00113##

17. The compound of claim 16, wherein R.sub.1 is propyl and R.sub.7 is acetyloxy.

18. The compound of claim 9, wherein the compound is of Formula (I-E) or (I-F), and R.sub.a is H, methyl, ethyl, prop-2-yn-1-yl, prop-2-en-1-yl, morpholino, benzyl, CH.sub.2CN, CH.sub.2CH.sub.2N(CH.sub.3).sub.2, or ##STR00114##

19. The compound of claim 1, wherein the compound is selected from Compounds 1-72.

20. A method of treating an inflammatory condition comprising administering to a subject in need thereof an effective amount of a compound of claim 1.

21. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.

Description

DETAILED DESCRIPTION

[0060] The present invention is based on a surprising discovery that the compounds of Formula (I) reproduced below are effective in inhibiting inflammation activity and treating disorders associated therewith.

##STR00084##

[0061] Variables R.sub.1-R.sub.7, X.sub.1, and X.sub.2 are defined above.

[0062] Formula (I) shows a bicyclo[8.4.0]tetradecane ring system as a core structure. The carbon atoms on the ring are numbered above for easy reference.

[0063] A subset of the compounds of Formula (I) is represented by Formula (I-A) as follows:

##STR00085##

[0064] Another subset of the compounds of Formula (I) is represented by Formula (I-B) as shown below:

##STR00086##

[0065] One more subset of the compounds of Formula (I) is represented by Formula (I-C):

##STR00087##

[0066] Still another subset of the compounds of Formula (I) is represented by Formula (I-D):

##STR00088##

[0067] Yet another subset of the compounds of Formula (I) is represented by Formula (I-E):

##STR00089##

[0068] Further subset of the compounds of Formula (I) is represented by Formula (I-F):

##STR00090##

[0069] One more subset of the compounds of Formula (I) is represented by Formula (I-G):

##STR00091##

[0070] In any of Formulas (I-A) to (I-G) above, variables R.sub.1, R.sub.7, R.sub.a, R.sub.b, R.sub.d, and R.sub.e are defined above.

[0071] The compounds of Formula (I) can be prepared by synthetic methods well known in the art. See, e.g., R. Larock, Comprehensive Organic Transformations (3.sup.rd Ed., John Wiley and Sons 2018); P. G. M. Wuts and T. W. Greene, Greene's Protective Groups in Organic Synthesis (4.sup.th Ed., John Wiley and Sons 2007); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis (John Wiley and Sons 1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis (2.sup.nd ed., John Wiley and Sons 2009) and subsequent editions thereof.

[0072] As illustrated below, these compounds can be prepared by one of the following Procedures A, B, and C.

##STR00092##

[0073] Provided below is an exemplary procedure. Under an ambient atmosphere (e.g., 20? C.), a 20-mL vial with a stir bar is charged with 1 molar equivalent Compound SP (e.g., ExcB, 182.7 mg, 0.307 mmol) and dried dichloromethane (2 mL), followed by the addition of pyridinium chlorochromate (PCC, 132.4 mg, 0.614 mmol, 2 equiv., commercially available from Millipore Sigma, Burlington, MA). The resultant mixture is stirred at room temperature (e.g., 20? C.) for 3 hours. It is then filtered through a short pad of diatomaceous earth (Celite?, MilliporeSigma) and rinsed with dichloromethane. The filtrate is collected and concentrated by rotary evaporation. The residue can be purified by flash column chromatography on silica gel (EtOAc:n-Hexane from 0:1 to 1:1) to afford Compound SP026 at a quantitative yield, usually as a white solid.

[0074] Compound 1 (1 equiv.) is then mixed with O-methylhydroxylamine hydrochloride (3.8 mg, 0.0449 mmol, 1.5 equiv.) in dichloromethane/pyridine (0.3 mL, ratio 14:1), and subsequently stirred at room temperature for 0.5 hours. Purification with flash column chromatography (EtOAc:n-Hexane from 0:1 to 1:1) affords a compound of Formula (I-A) at an excellent yield (e.g., 90% or greater), often as a colorless oil.

[0075] The following compounds were prepared using Procedure A: Compounds 1, 4, 5, 40, and 67.

[0076] Compound 1: .sup.1H NMR (600 MHz, CDCl.sub.3, 22? C.) ? 5.78 (s, 1H), 5.35 (s, 2H), 5.31-5.23 (m, 2H), 5.17 (dd, J=9.8, 4.0 Hz, 1H), 3.61 (dd, J=15.7, 4.9 Hz, 1H), 3.25 (dd, J=9.3, 5.7 Hz, 1H), 3.01-2.93 (m, 1H), 2.86 (t, J=13.3 Hz, 1H), 2.78 (dd, J=16.5, 4.0 Hz, 1H), 2.31 (s, 3H), 2.24 (t, J=7.4 Hz, 2H), 2.18 (s, 3H), 2.11 (s, 3H), 2.10-2.06 (m, 1H), 1.81 (s, 3H), 1.66-1.60 (m, 2H), 1.59 (s, 3H), 1.23 (d, J=7.5 Hz, 3H), 1.14 (s, 3H), 0.94 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.30H.sub.40O.sub.12Na [M+Na].sup.+ 615.2412, found 615.2416.

[0077] Compound 4: HRMS-FAB (m/z) calculated for C.sub.30H.sub.41NO.sub.12Na [M+Na].sup.+ 630.2521, found 630.2514.

[0078] Compound 5: HRMS-ESI (m/z) calculated for C.sub.31H.sub.43NO.sub.12Na [M+Na].sup.+ 644.2678, found 644.2669.

[0079] Compound 40: MS-ESI (m/z) calculated for C.sub.33H.sub.44NO.sub.12 [M+H].sup.+ 646.2864, found 646.3.

[0080] Compound 67: HRMS-FAB (m/z) calculated for C.sub.30H.sub.41NO.sub.12Na [M+Na]+ 630.2521, found 630.2514.

##STR00093##

[0081] At room temperature, a 4-mL vial equipped with a stir bar is charged with 1 molar equivalent Compound SP001 (e.g., ExcB, 182.7 mg, 0.307 mmol), SeO.sub.2 (100.9 mg, 0.91 mmol, 10 equiv., MilliporeSigma), and 1,4-dioxane (1 mL, MilliporeSigma). The resultant mixture is stirred at 80? C. for 22 hours, cooled to room temperature, and then filtered through a short pad of Celite?. The filtrate is concentrated under reduced pressure to give a residue, which is purified by flash column chromatography (EtOAc:n-Hexane from 0:1 to 2:3 then to 1:1) to afford Compound SP038 at a high yield (e.g., 80% or greater), usually as a white solid.

[0082] Compound SP038 (1 equiv.) is then mixed with 1.5 equivalents O-methylhydroxylamine hydrochloride in dichloromethane/pyridine (0.3 mL, ratio 14:1), and subsequently stirred at room temperature for 0.5 hours. Purification with flash column chromatography (EtOAc:n-Hexane from 0:1 to 1:1) affords a compound of Formula I-B at a high yield (e.g., 80% or greater) often as a colorless oil.

[0083] The following compounds were prepared using Procedure B: Compounds 7, 37, 41, 51, and 56-60.

[0084] Compound 7: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 9.69 (d, J=1.6 Hz, 1H), 6.88 (dd, J=7.5, 2.1 Hz, 1H), 5.96 (dd, J=7.4, 1.5 Hz, 1H), 5.71 (dd, J=7.6, 2.7 Hz, 1H), 5.64 (d, J=10.4 Hz, 1H), 5.19 (d, J=2.5 Hz, 1H), 4.59-4.51 (m, 1H), 4.20-4.11 (m, 1H), 3.96-3.79 (m, 2H), 2.87 (dd, J=10.4, 5.0 Hz, 1H), 2.57-2.50 (m, 1H), 2.46 (d, J=15.1 Hz, 1H), 2.42 (s, 3H), 2.21 (s, 3H), 2.22-2.05 (m, 2H), 1.99 (s, 3H), 1.89-1.81 (m, 1H), 1.79-1.71 (m, 1H), 1.55 (s, 3H), 1.54-1.49 (m, 2H), 1.03 (d, J=7.2 Hz, 3H), 0.90 (t, J=7.4 Hz, 3H), 0.79 (s, 3H). HRMS-ESI (m/z) calculated for C.sub.30H.sub.40O.sub.13Na [M+Na].sup.+ 631.2361, found 631.2370.

[0085] Compound 37: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 7.97 (s, 1H), 6.06-5.99 (m, 1H), 5.92 (d, J=6.0 Hz, 1H), 5.83 (d, J=7.4 Hz, 1H), 5.61 (d, J=10.3 Hz, 1H), 5.17 (s, 1H), 4.58 (s, 1H), 4.22-4.15 (m, 1H), 3.97 (dd, J=15.7, 7.3 Hz, 1H), 3.91-3.83 (m, 1H), 3.75 (s, 3H), 2.98 (dd, J=10.4, 5.0 Hz, 1H), 2.57-2.48 (m, 2H), 2.41 (s, 3H), 2.21 (s, 3H), 2.19-2.08 (m, 2H), 2.05 (s, 3H), 1.91-1.80 (m, 1H), 1.80-1.73 (m, 1H), 1.58-1.46 (m, 2H), 1.53 (s, 3H), 1.04 (d, J=7.2 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H), 0.81 (s, 3H). HRMS-ESI (m/z) calculated for C.sub.31H.sub.43NO.sub.13Na [M+Na].sup.+ 660.2627, found 660.2634.

[0086] Compound 41: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 8.04 (s, 1H), 6.09 (dd, J=7.5, 2.0 Hz, 1H), 5.92 (dd, J=6.6, 2.0 Hz, 1H), 5.84 (d, J=7.4 Hz, 1H), 5.62 (d, J=10.4 Hz, 1H), 5.16 (d, J=2.5 Hz, 1H), 4.65 (dd, J=15.8, 2.5 Hz, 1H), 4.61-4.55 (m, 2H), 4.17 (d, J=4.0 Hz, 1H), 4.01 (dd, J=15.6, 6.7 Hz, 1H), 3.91-3.84 (m, 1H), 3.26 (t, J=2.4 Hz, 1H), 2.96 (dd, J=10.4, 5.1 Hz, 1H), 2.58-2.49 (m, 2H), 2.41 (s, 3H), 2.21 (s, 3H), 2.17-2.05 (m, 2H), 2.05 (s, 3H), 1.90-1.80 (m, 1H), 1.79-1.72 (m, 1H), 1.58-1.48 (m, 2H), 1.53 (s, 3H), 1.04 (d, J=7.3 Hz, 3H), 0.89 (t, J=6.0 Hz, 3H), 0.81 (s, 3H). HRMS-ESI (m/z) calculated for C.sub.33H.sub.43NO.sub.13Na [M+Na].sup.+ 684.2627, found 684.2620.

[0087] Compound 51: MS-ESI (m/z) calcd for C.sub.34H.sub.49N.sub.2O.sub.13 [M+H].sup.+ 693.3235, found 693.3.

[0088] Compound 56: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 10.81 (s, 1H), 7.97 (s, 1H), 5.98 (d, J=7.6 Hz, 1H), 5.91 (d, J=4.9 Hz, 1H), 5.83 (d, J=7.7 Hz, 1H), 5.60 (d, J=10.3 Hz, 1H), 5.21 (br s, 1H), 4.56 (br s, 1H), 4.19 (d, J=3.9 Hz, 1H), 3.92 (dd, J=15.4, 7.2 Hz, 1H), 3.89-3.84 (m, 1H), 3.03 (dd, J=10.4, 5.0 Hz, 1H), 2.57-2.49 (m, 2H), 2.41 (s, 3H), 2.20 (s, 3H), 2.14-2.05 (m, 1H), 2.02 (s, 3H), 2.03-1.93 (m, 1H), 1.89-1.80 (m, 1H), 1.79-1.73 (m, 1H), 1.52 (s, 3H), 1.51-1.40 (m, 2H), 1.03 (d, J=7.1 Hz, 3H), 0.84 (t, J=7.4 Hz, 3H), 0.80 (s, 3H). HRMS-ESI (m/z) calculated for C.sub.30H.sub.41NO.sub.13Na [M+Na].sup.+ 646.2470, found 646.2461.

[0089] Compound 57: MS-ESI (m/z) calculated for C.sub.30H.sub.41NO.sub.13Na [M+Na].sup.+ 646.2470, found 646.2461.

[0090] Compound 58: MS-ESI (m/z) calculated for C.sub.32H.sub.46NO.sub.13 [M+H].sup.+ 652.2969, found 652.3.

[0091] Compound 59: MS-ESI (m/z) calculated for C.sub.33H.sub.46NO.sub.13 [M+H].sup.+ 664.2969, found 664.3.

[0092] Compound 60: MS-ESI (m/z) calculated for C.sub.32H.sub.42N.sub.2O.sub.13Na [M+Na].sup.+ 685.2585, found 685.2.

##STR00094##

##STR00095##

##STR00096##

[0093] Schemes III, IV, and V above show preparation of a compound of one of Formula (I-D), (I-E), and (I-F). At room temperature, 1 molar equivalent Compound SP001 or a compound of Formula (I-B) or (I-C) is mixed with carboxylic acid R.sub.dCOOH (2 equiv., MilliporeSigma), dimethylamino-pyridine (DMAP, 0.3 equiv., MilliporeSigma), 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide (EDCI, 3 equiv., MilliporeSigma) and dried CH.sub.2Cl.sub.2 (0.5 mL) or dried dimethyl-formamide (0.5 mL, MilliporeSigma). The resultant mixture is stirred at room temperature for 18 hours, diluted with CH.sub.2Cl.sub.2, and then washed with water. The organic layer is collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of corresponding Formula (I-D), (I-E), or (I-F).

[0094] The following compounds were prepared using Procedure C: Compounds 8, 15, 16, 17, 21-27, 29-36, 42, 44, 45, 47, 48, 52, 55, 64, 65, 66, and 68.

[0095] Compound 8: MS-ESI (m/z) calculated for C.sub.35H.sub.46O.sub.10Na [M+Na].sup.+ 697.2836, found 697.2.

[0096] Compound 15: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 6.75 (X part of ABX system, J=6.3 Hz, 1H), 5.85 (dd, J=6.9, 1.8 Hz, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.53 (d, J=10.4 Hz, 1H), 5.36 (dt, J=7.4, 1.9 Hz, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.01 (ddd, J=12.5, 5.0, 3.6 Hz, 1H), 4.74-4.66 (m, 1H), 4.01 (dd, J=15.8, 7.3 Hz, 1H), 3.81, 3.78 (AB part of ABX system, J.sub.AB=18.0 Hz, J.sub.AX=6.6 Hz, J.sub.BX=6.6 Hz, 2H), 3.17 (dd, J=10.4, 5.1 Hz, 1H), 2.68-2.60 (m, 1H), 2.41 (s, 3H), 2.23 (s, 6H), 2.22-2.06 (m, 4H), 1.95 (s, 3H), 1.90-1.85 (m, 1H), 1.55-1.46 (m, 2H), 1.50 (s, 3H), 1.37 (s, 9H), 1.10 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.37H.sub.53NO.sub.15Na [M+Na].sup.+ 774.3307, found 774.3311.

[0097] Compound 16: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.85 (dd, J=6.9, 1.7 Hz, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.52 (d, J=10.4 Hz, 1H), 5.40-5.33 (m, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.03 (ddd, J=12.4, 5.0, 3.6 Hz, 1H), 4.74-4.67 (m, 1H), 4.01 (dd, J=15.8, 7.3 Hz, 1H), 3.27, 3.24 (ABq, J=16.9 Hz, 2H), 3.16 (dd, J=10.4, 5.1 Hz, 1H), 2.67-2.60 (m, 1H), 2.41 (s, 3H), 2.30 (s, 6H), 2.23 (s, 3H), 2.23 (s, 3H), 2.22-2.07 (m, 4H), 1.95 (s, 3H), 1.91-1.85 (m, 1H), 1.55-1.46 (m, 2H), 1.50 (s, 3H), 1.09 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.34H.sub.50NO.sub.13 [M+H].sup.+ 680.3277, found 680.3279.

[0098] Compound 17: HRMS-ESI (m/z) calculated for C.sub.32H.sub.46NO.sub.13 [M+H].sup.+ 652.2964, found 652.2966.

[0099] Compound 21: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.88-5.82 (m, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.52 (d, J=10.4 Hz, 1H), 5.37 (dt, J=7.4, 1.8 Hz, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.02 (ddd, J=12.5, 5.0, 3.6 Hz, 1H), 4.70 (dd, J=4.1, 2.1 Hz, 1H), 4.01 (dd, J=15.8, 7.3 Hz, 1H), 3.79-3.40 (m, 4H), 3.24, 3.21 (ABq, J=17.0 Hz, 2H), 3.16 (dd, J=10.4, 5.2 Hz, 1H), 2.79-2.59 (m, 3H), 2.41 (s, 3H), 2.38-2.26 (m, 2H), 2.23 (s, 3H), 2.23 (s, 3H), 2.21-2.06 (m, 4H), 1.95 (s, 3H), 1.90-1.85 (m, 1H), 1.56-1.46 (m, 2H), 1.51 (s, 3H), 1.09 (d, J=7.2 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.36H.sub.52NO.sub.14 [M+H].sup.+ 722.3382, found 722.3388.

[0100] Compound 22: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.85 (d, J=7.6 Hz, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.52 (d, J=10.4 Hz, 1H), 5.36 (dd, J=7.3, 2.0 Hz, 1H), 5.16 (d, J=2.3 Hz, 1H), 5.07-4.98 (m, 1H), 4.70 (br s, 1H), 4.01 (dd, J=15.8, 7.3 Hz, 1H), 3.21, 3.18 (ABq, J=16.9 Hz, 2H), 3.17 (dd, J=10.6, 5.2 Hz, 1H), 2.77-2.66 (m, 2H), 2.68-2.58 (m, 3H), 2.41 (s, 3H), 2.30-2.15 (m, 3H), 2.23 (s, 6H), 2.14-2.01 (m, 3H), 2.12 (s, 3H), 1.99-1.91 (m, 2H) 1.95 (s, 3H), 1.91-1.84 (m, 1H), 1.53-1.47 (m, 2H), 1.50 (s, 3H), 1.09 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.3 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.37H.sub.55N.sub.2O.sub.13 [M+H].sup.+ 735.3699, found 735.3694.

[0101] Compound 23: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.88-5.82 (m, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.53 (d, J=10.3 Hz, 1H), 5.36 (d, J=7.3 Hz, 1H), 5.16 (d, J=2.4 Hz, 1H), 4.99 (ddd, J=12.4, 5.0, 3.6 Hz, 1H), 4.72-4.68 (m, 1H), 4.01 (dd, J=15.8, 7.2 Hz, 1H), 3.15 (dd, J=10.3, 5.1 Hz, 1H), 2.71-2.58 (m, 3H), 2.55-2.41 (m, 6H), 2.41 (s, 3H), 2.23 (s, 6H), 2.21-2.07 (m, 3H), 2.04-2.00 (m, 1H), 1.95 (s, 3H), 1.89-1.84 (m, 1H), 1.71-1.63 (m, 4H), 1.57-1.46 (m, 2H), 1.50 (s, 3H), 1.10 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.3 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.37H.sub.54NO.sub.13 [M+H].sup.+ 720.3590, found 720.3598.

[0102] Compound 24: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.88-5.82 (m, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.53 (d, J=10.3 Hz, 1H), 5.16 (d, J=2.3 Hz, 1H), 5.38-5.34 (m, 1H), 5.00 (dt, J=12.4, 4.3 Hz, 1H), 4.70 (dd, J=3.9, 2.1 Hz, 1H), 4.73-4.67 (m, 1H), 4.01 (dd, J=15.8, 7.2 Hz, 1H), 3.14 (dd, J=10.3, 5.1 Hz, 1H), 2.94-2.80 (m, 2H), 2.69-2.61 (m, 1H), 2.57-2.42 (m, 4H), 2.41 (s, 3H), 2.23 (s, 3H), 2.23 (s, 3H), 2.22-2.00 (m, 5H), 1.95 (s, 3H), 1.90-1.83 (m, 1H), 1.81-1.74 (m, 2H), 1.66-1.35 (m, 7H), 1.50 (s, 3H), 1.04-1.17 (m, 1H), 1.10 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.38H.sub.56NO.sub.13 [M+H].sup.+ 734.3746, found 734.3741.

[0103] Compound 25: MS-ESI (m/z) calcd for C.sub.34H.sub.48NO.sub.14[M+H].sup.+ 694.3075, found 694.3.

[0104] Compound 26: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 8.52-8.46 (m, 1H), 7.77 (td, J=7.7, 1.8 Hz, 1H), 7.39 (d, J=7.8 Hz, 1H), 7.28 (ddd, J=7.6, 4.9, 1.1 Hz, 1H), 5.84 (dd, J=6.9, 1.8 Hz, 1H), 5.66 (d, J=7.3 Hz, 1H), 5.52 (d, J=10.4 Hz, 1H), 5.39-5.33 (m, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.02 (ddd, J=12.4, 5.0, 3.6 Hz, 1H), 4.72-4.67 (m, 1H), 4.00 (dd, J=15.8, 7.3 Hz, 1H), 3.87, 3.84 (ABq, J=16.0 Hz, 2H), 3.16 (dd, J=10.4, 5.2 Hz, 1H), 2.68-2.60 (m, 1H), 2.40 (s, 3H), 2.22 (s, 3H), 2.19 (s, 3H), 2.19-2.06 (m, 2H), 2.03-1.99 (m, 1H), 1.94 (s, 3H), 1.91-1.86 (m, 1H), 1.55-1.46 (m, 2H), 1.51 (s, 3H) 1.06 (d, J=7.1 Hz, 3H), 0.85 (s, 3H), 0.85 (t, J=7.5 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.37H.sub.47NO.sub.13Na [M+Na].sup.+ 736.2940, found 736.2942.

[0105] Compound 27: MS-ESI (m/z) calcd for C.sub.40H.sub.54NO.sub.13[M+H].sup.+ 756.3595, found 756.3.

[0106] Compound 29: HRMS-ESI (m/z) calcd for C.sub.36H.sub.54NO.sub.13 [M+H].sup.+ 708.3590, found 708.3594.

[0107] Compound 30: MS-ESI (m/z) calcd for C.sub.38H.sub.58NO.sub.13[M+H].sup.+ 736.3980, found 736.4.

[0108] Compound 31: HRMS-ESI (m/z) calcd for C.sub.33H.sub.48NO.sub.13 [M+H].sup.+ 666.3120, found 666.3117.

[0109] Compound 32: MS-ESI (m/z) calcd for C.sub.35H.sub.52NO.sub.13[M+H].sup.+ 694.3439, found 694.3.

[0110] Compound 33: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.85 (dd, J=7.2, 1.4 Hz, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.53 (d, J=10.3 Hz, 1H), 5.41-5.34 (m, 1H), 5.16 (d, J=2.3 Hz, 1H), 5.01 (ddd, J=12.5, 5.0, 3.6 Hz, 1H), 4.70 (dd, J=4.0, 2.1 Hz, 1H), 4.01 (dd, J=15.9, 7.2 Hz, 1H), 3.68 (br s, 2H), 3.36 (br s, 2H), 3.15 (dd, J=10.4, 5.1 Hz, 1H), 2.74-2.61 (m, 3H), 2.56-2.51 (m, 2H), 2.51-2.44 (m, 2H), 2.41 (s, 3H), 2.23 (s, 6H), 2.21-2.02 (m, 4H), 2.02-1.98 (m, 1H), 1.96 (s, 3H), 1.90-1.84 (m, 1H), 1.56-1.45 (m, 2H), 1.51 (s, 3H), 1.10 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.37H.sub.54NO.sub.14 [M+H].sup.+ 736.3539, found 736.3545.

[0111] Compound 34: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.85 (dd, J=7.3, 1.3 Hz, 1H), 5.67 (d, J=7.2 Hz, 1H), 5.53 (d, J=10.3 Hz, 1H), 5.37 (dt, J=7.4, 1.7 Hz, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.00 (ddd, J=12.4, 5.0, 3.6 Hz, 1H), 4.70 (dd, J=4.0, 2.1 Hz, 1H), 4.02 (dd, J=15.9, 7.2 Hz, 1H), 3.14 (dd, J=10.4, 5.1 Hz, 1H), 2.77-2.70 (m, 2H), 2.69-2.59 (m, 3H), 2.57-2.42 (m, 4H), 2.41 (s, 3H), 2.23 (s, 6H), 2.21-2.06 (m, 3H), 2.13 (s, 3H), 2.03-1.89 (m, 5H), 1.95 (s, 3H), 1.89-1.82 (m, 1H), 1.56-1.46 (m, 2H), 1.51 (s, 3H), 1.10 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.38H.sub.57N.sub.2O.sub.13 [M+H].sup.+ 749.3855, found 749.3864.

[0112] Compound 35: HRMS-ESI (m/z) calcd for C.sub.36H.sub.52NO.sub.13 [M+H].sup.+ 706.3433, found 706.3433.

[0113] Compound 36: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.88-5.82 (m, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.52 (d, J=10.4 Hz, 1H), 5.40-5.32 (m, 1H), 5.16 (d, J=2.3 Hz, 1H), 4.98 (dt, J=12.4, 4.3 Hz, 1H), 4.70 (dd, J=4.0, 2.1 Hz, 1H), 4.01 (dd, J=15.8, 7.2 Hz, 1H), 3.15 (dd, J=10.4, 5.1 Hz, 1H), 2.69-2.57 (m, 1H), 2.41 (s, 3H), 2.30 (td, J=7.6, 4.7 Hz, 2H), 2.23 (s, 3H), 2.22 (s, 3H), 2.20-1.99 (m, 4H), 1.95 (s, 3H), 1.89-1.82 (m, 1H), 1.73-1.57 (m, 6H), 1.57-1.45 (m, 2H), 1.50 (s, 3H), 1.45-1.37 (m, 2H), 1.21-1.06 (m, 3H), 1.09 (d, J=7.1 Hz, 3H), 0.90-0.77 (m, 2H), 0.86 (s, 3H), 0.85 (t, J=7.5 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.39H.sub.56O.sub.13Na [M+H].sup.+ 755.3613, found 755.3621.

[0114] Compound 42: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.87 (dd, J=7.6, 2.6 Hz, 1H), 5.68 (d, J=7.3 Hz, 1H), 5.54 (d, J=10.3 Hz, 1H), 5.42-5.35 (m, 1H), 5.18 (d, J=2.4 Hz, 1H), 5.04 (dt, J=12.5, 4.3 Hz, 1H), 4.72 (dd, J=4.0, 2.1 Hz, 1H), 4.03 (dd, J=15.8, 7.3 Hz, 1H), 3.29 (d, J=2.5 Hz, 2H), 3.26, 3.20 (ABq, J=17.0 Hz, 2H), 3.18 (dd, J=17.2, 4.9 Hz, 1H), 2.99-2.93 (m, 1H), 2.86-2.75 (m, 2H), 2.73-2.60 (m, 3H), 2.42 (s, 3H), 2.37-2.27 (m, 4H), 2.24 (s, 6H), 2.22-2.08 (m, 4H), 1.97 (s, 3H), 1.92-1.86 (m, 1H), 1.58-1.47 (m, 2H), 1.52 (s, 3H), 1.11 (d, J=7.1 Hz, 3H), 0.87 (s, 3H), 0.87 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.39H.sub.55N.sub.2O.sub.13 [M+H].sup.+ 759.3699, found 759.3701.

[0115] Compound 44: HRMS-ESI (m/z) calcd for C.sub.38H.sub.56NO.sub.13 [M+H].sup.+ 734.3746, found 734.3741.

[0116] Compound 45: H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.88-5.80 (m, 1H), 5.67 (d, J=7.3 Hz, 1H), 5.53 (d, J=10.3 Hz, 1H), 5.41-5.33 (m, 1H), 5.16 (d, J=2.4 Hz, 1H), 5.00 (ddd, J=12.5, 5.1, 3.7 Hz, 1H), 4.75-4.65 (m, 1H), 4.01 (dd, J=15.8, 7.2 Hz, 1H), 3.15 (dd, J=10.4, 5.1 Hz, 1H), 2.72 (br s, 2H), 2.66 (td, J=7.0, 5.2 Hz, 1H), 2.57 (br s, 4H), 2.44 (br s, 2H), 2.41 (s, 3H), 2.23 (s, 3H), 2.22 (s, 3H), 2.21-2.02 (m, 4H), 1.95 (s, 3H), 1.92-1.84 (m, 1H), 1.61-1.45 (m, 10H), 1.50 (s, 3H), 1.11 (d, J=7.1 Hz, 3H), 0.86 (s, 3H), 0.85 (t, J=7.3 Hz, 3H). HRMS-ESI (m/z) calcd for C.sub.39H.sub.58NO.sub.13 [M+H].sup.+ 748.3903, found 748.3900.

[0117] Compound 47: HRMS-ESI (m/z) calcd for C.sub.39H.sub.58NO.sub.13 [M+H].sup.+ 748.3903, found 748.3897.

[0118] Compound 48: HRMS-ESI (m/z) calcd for C.sub.40H.sub.60NO.sub.13 [M+H].sup.+ 762.4059, found 762.4049.

[0119] Compound 52: MS-ESI (m/z) calcd for C.sub.38H.sub.54NO.sub.14 [M+H].sup.+ 748.3544, found 748.3.

[0120] Compound 55: MS-ESI (m/z) calcd for C.sub.39H.sub.57N.sub.2O.sub.14 [M+H].sup.+, 777.3810 found 777.4.

[0121] Compound 64: MS-ESI (m/z) calcd for C.sub.38H.sub.56NO.sub.13 [M+H].sup.+ 734.3752, found 734.4.

[0122] Compound 65: MS-ESI (m/z) calculated for C.sub.39H.sub.57N.sub.2O.sub.15 [M+H].sup.+ 793.3759, found 793.4.

[0123] Compound 66: MS-ESI (m/z) calcd for C.sub.41H.sub.57N.sub.2O.sub.14 [M+H].sup.+ 801.3810, found 801.4.

[0124] Compound 68: MS-ESI (m/z) calcd for C.sub.41H.sub.57N.sub.3O.sub.14 [M+H].sup.+ 816.3919, found 816.4.

Procedure D

[0125] Compounds of Formula (I-C1) can be prepared following the procedure illustrated in Scheme VI below.

##STR00097##

[0126] Compound 7 (100 mg, 0.164 mmol) is dissolved in t-butanol (0.8 mL), THF (0.8 mL), and 2-methyl-2-butene (0.3 mL, 3.28 mmol). A mixture of NaClO.sub.2 (81.5 mg, 0.902 mmol) and NaH.sub.2PO.sub.4 (127.7 mg, 1.07 mmol) in water (0.3 mL) is then added to the Compound 7 solution. After being stirred at room temperature for 20 hrs, the reaction mixture is quenched with 1 N HCl (2 mL). Ethyl acetate is added. The organic layer is collected. The aqueous layer is extracted with ethyl acetate. The organic layers are combined, dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford Compound 28.

[0127] At room temperature, Compound 28 (1 equiv.) is mixed with amines (1.1 equiv., MilliporeSigma), 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU, 1.5 equiv., MilliporeSigma), diisopropylethylamine (3 equiv.) and dried DMF (0.5 mL) or dried dichloromethane (0.5 mL, MilliporeSigma). The resultant mixture is stirred at room temperature for 20 hours, diluted with ethyl acetate, and then washed with water. The organic layer is collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of Formula (I-C1).

[0128] The following compounds were prepared using Procedure D: Compounds 28 and 61-63.

[0129] Compound 28: HRMS-ESI (m/z) calcd for C.sub.30H.sub.40O.sub.14Na [M+Na].sup.+ 647.2310, found 647.2311.

[0130] Compound 61: MS-ESI (m/z) calcd for C.sub.34H.sub.51N.sub.2O.sub.14 [M+H].sup.+ 711.3340, found 711.4.

[0131] Compound 62: MS-ESI (m/z) calcd for C.sub.31H.sub.44NO.sub.14 [M+H].sup.+ 654.2762, found 654.3.

[0132] Compound 63: MS-ESI (m/z) calcd for C.sub.33H.sub.44NO.sub.14 [M+H].sup.+ 678.2762, found 678.3.

Procedure E

[0133] Compounds of Formula (I-A1) can be prepared following the procedure illustrated in Scheme VII below.

##STR00098##

[0134] At room temperature, Compound 1 (1 molar equiv.) is mixed with hydrazine (1.5 equiv., MilliporeSigma), AcOH (2 drops) and toluene (0.3 mL). The resultant mixture is stirred at reflux for 20 hours, diluted with ethyl acetate, and then washed with water. The organic layer is collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of Formula (I-A1).

[0135] The following compounds were prepared using Procedure E: Compounds 9-12 and 18-20.

[0136] Compound 9: MS-ESI (m/z) calcd for C.sub.32H.sub.45O.sub.20 [M+H].sup.+ 617.3407, found 617.3.

[0137] Compound 10: MS-ESI (m/z) calcd for C.sub.33H.sub.48O.sub.39 [M+H].sup.+ 630.3391, found 630.3.

[0138] Compound 11: MS-ESI (m/z) calcd for C.sub.32H.sub.43O.sub.20 [M+H].sup.+ 615.2918, found 615.3.

[0139] Compound 12: MS-ESI (m/z) calcd for C.sub.32H.sub.43O.sub.20 [M+H].sup.+ 615.2918, found 615.3.

[0140] Compound 18: MS-ESI (m/z) calcd for C.sub.30H.sub.41O.sub.20 [M+H].sup.+ 589.2761, found 589.3.

[0141] Compound 19: MS-ESI (m/z) calcd for C.sub.30H.sub.41O.sub.20 [M+H].sup.+ 589.2761, found 589.3.

[0142] Compound 20: MS-ESI (m/z) calcd for C.sub.35H.sub.43O.sub.20 [M+H].sup.+ 651.2918, found 651.3.

Procedure F

[0143] Scheme VIII below depicts exemplary preparation of compounds of formula (I-H) of this invention.

##STR00099##

[0144] At room temperature, SP001 or SP040 (1 molar equiv.) is mixed with sodium azide (4 equiv., MilliporeSigma), DMF (0.3 mL). The resultant mixture is stirred at 80? C. for 16 hours, quenched with water, and then extracted with ethyl acetate. The combined organic layers were dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of Formula.

[0145] The following compounds were prepared using Procedure F: Compounds 2, 38 and 43.

[0146] Compound 2: HRMS-ESI (m/z) calculated for C.sub.30H.sub.41N.sub.3O.sub.11Na [M+Na].sup.+ 642.2633, found 642.2633.

[0147] Compound 38: HRMS-FAB (m/z) calculated for C.sub.30H.sub.44N.sub.3O.sub.12 [M+H].sup.+ 638.2925, found 638.2921.

[0148] Compound 43: MS-ESI (m/z) calculated for C.sub.40H.sub.55N.sub.4O.sub.13 [M+H].sup.+ 799.3766, found 799.4.

Procedure G

[0149] Scheme IX below illustrates a method for preparing compounds of formula (I-I) of this invention.

##STR00100##

[0150] To a stirring solution of diphenyl diselenide (1.5 equiv.) in EtOH (0.6 mL) under a nitrogen atmosphere was added NaBH.sub.4 (3 equiv.) at room temperature. After evolution of hydrogen ceased, the light-yellow solution was cooled to 0? C., followed by the addition of acetic acid (1.5 equiv.), and the mixture was further stirred at 0? C. for 50 min. The resulting colorless solution was then added to a solution of SP001 (1 equiv.) in EtOH (0.6 mL) at room temperature. The reaction mixture was diluted with ethyl acetate, and oxygen was passed through the solution for 5 minutes. The reaction mixture was washed with H.sub.2O. The organic layer was collected, dried over MgSO.sub.4, filtered, and concentrated by rotary evaporation. The residue was purified by flash column chromatography to afford a compound of Formula.

[0151] The following compounds were prepared using Procedure G: Compound 54.

[0152] Compound 54: HRMS-ESI (m/z) calculated for C.sub.30H.sub.44NO.sub.12Na [M+Na].sup.+ 619.2725, found 619.2735.

Procedure H

[0153] Scheme X below shows an exemplary method of preparing compound 3 and its analogs of this invention.

##STR00101##

[0154] At room temperature, SP001 (1 equiv.) is mixed with CBr.sub.4 (3.0 equiv., MilliporeSigma), PPh.sub.3 (3.0 equiv., MilliporeSigma) and dried THF (4 mL). The resultant mixture is stirred at room temperature for 16 hours, the reaction mixture was filtered through a short pad of Celite to remove triphenylphosphine oxide and rinsed with ether. The filtrate was concentrated and purified by prep HPLC (C18) to afford a compound of Formula.

[0155] Compound 3 was prepared using Procedure H: HRMS-FAB (m/z) calculated for C.sub.30H.sub.42.sup.79BrO.sub.11 [M+H].sup.+ 657.1910, found 657.1911; C.sub.30H.sub.42.sup.81BrO.sub.11 [M+H].sup.+ 659.1890, found 659.1880.

[0156] SP040 was prepared using Procedure H: HRMS-ESI (m/z) calculated for C.sub.30H.sub.40O.sub.11Na [M+Na].sup.+ 599.2463, found 599.2465.

Procedure I

[0157] Scheme XI below illustrates a method of preparing compounds of formula (I-J) of this invention.

##STR00102##

[0158] At room temperature, SP001 or compound 7 (1 molar equiv.) is mixed with sodium borohydride (2 equiv., MilliporeSigma), and methanol (0.3 mL). The resultant mixture is stirred at 0? C. for 3 hours, quenched with water, and then extracted with ethyl acetate. The combined organic layers were dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of Formula.

[0159] The following compounds were prepared using Procedure I: Compounds 39 and 46.

[0160] Compound 39: MS-ESI (m/z) calculated for C.sub.30H.sub.44NaO.sub.13 [M+Na].sup.+ 635.2680, found 635.3.

[0161] Compound 46: HRMS-ESI (m/z) calculated for C.sub.30H.sub.44O.sub.12Na [M+Na].sup.+ 619.2725, found 619.2734.

Procedure J

[0162] Scheme XII below illustrates preparation of compounds of formula (I-G) of this invention.

##STR00103##

[0163] At room temperature, SP001 is mixed with RSO.sub.2Cl (2 equiv.), triethylamine (3 equiv., MilliporeSigma) and dried DCM (0.4 mL). The resultant mixture is stirred at room temperature for 16 hours, quenched with water, and then extracted with DCM. The combined organic layers were dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by flash column chromatography to afford a compound of Formula.

[0164] The following compounds were prepared using Procedure J: Compounds 13, 14 and 49.

[0165] Compound 13: MS-ESI (m/z) calculated for C.sub.30H.sub.43NO.sub.20SNa[M+Na].sup.+ 696.2302, found 696.2.

[0166] Compound 14: MS-ESI (m/z) calculated for C.sub.31H.sub.45O.sub.14S[M+H].sup.+ 673.2530, found 673.2.

[0167] Compound 49: .sup.1H NMR (600 MHz, acetone-d.sub.6, ?40? C.) ? 5.80 (dd, J=7.3, 1.7 Hz, 1H), 5.66 (d, J=7.2 Hz, 1H), 5.55 (d, J=10.2 Hz, 1H), 5.32 (d, J=7.3 Hz, 1H), 5.16 (d, J=2.3 Hz, 1H), 4.94 (dt, J=12.6, 4.1 Hz, 1H), 4.76-4.72 (m, 1H), 4.00 (dd, J=15.9, 7.0 Hz, 1H), 3.45-3.33 (m, 2H), 3.18 (dd, J=10.2, 5.1 Hz, 1H), 2.96-2.76 (m, 3H), 2.65 (hept, J=7.1, 6.4 Hz, 2H), 2.41 (s, 3H), 2.34-2.28 (m, 1H), 2.25 (s, 3H), 2.23 (s, 3H), 2.22-2.06 (m, 4H), 1.97-1.84 (m, 2H), 1.91 (s, 3H), 1.61-1.39 (m, 11H), 1.15 (d, J=7.1 Hz, 3H), 0.87 (s, 3H), 0.85 (t, J=7.4 Hz, 3H). HRMS-ESI (m/z) calculated for C.sub.37H.sub.56NO.sub.14S [M+H].sup.+ 770.3416, found 770.3410.

Procedure K

[0168] Scheme XIII below shows a method of preparing Compound 6 and its analogs of this invention.

##STR00104##

[0169] At room temperature, Compound 38 (1 equiv.) was mixed with PPh.sub.3 (4 equiv.) and THF/H.sub.2O (40:1, 1.2 mL). The resultant mixture was stirred at room temperature for 16 hours, quenched with water, and then extracted with ethyl acetate. The combined organic layers were dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which was purified by flash column chromatography to afford Compound 6.

[0170] MS-ESI (m/z) calculated for C.sub.30H.sub.42NO.sub.10[M+H].sup.+ 576.2809, found 576.3.

Procedure L

[0171] Scheme XIV below shows a method of preparing Compound 50 and it analogs of this invention.

##STR00105##

[0172] At room temperature, SP001 (1 equiv.) was mixed with Pd/C and methanol (1.5 mL). The resultant mixture was stirred under H.sub.2 (balloon) at room temperature for 16 hours, filtered through a pad of Celite, and then rinsed with ethyl acetate. The filtrate was concentrated to obtain a residue, which was purified by flash column chromatography to afford compound 50: MS-ESI (m/z) calculated for C.sub.30H.sub.45O.sub.12 [M+H].sup.+ 597.2911, found 597.3.

Procedure M

[0173] Compound 53 was prepared following the method shown in Scheme XV below.

##STR00106##

[0174] At room temperature, compound 41 (1 equiv.) was mixed with S1 (1 equiv.), CuSO.sub.4 5H.sub.2O (0.2 equiv.), sodium ascorbate (2 equiv.) and MeOH (0.6 mL). The resultant mixture was stirred at room temperature for 20 min, filtered through a short pad of Celite and rinsed with ethyl acetate. The filtrate was collected and concentrated to obtain a residue, which was purified by flash column chromatography to afford compound 53.

[0175] MS-ESI (m/z) calculated for C.sub.56H.sub.72N.sub.7O.sub.22 [M+H].sup.+ 1194.4730, found 1194.5.

[0176] Compound 69 was prepared following the method shown in Scheme XVI below.

##STR00107##

[0177] At room temperature, Compound SP001 (1 equiv.) was mixed with (E)-4-(tert-butoxy)-4-oxobut-2-enoic acid (1.2 equiv.), dimethylamino-pyridine (DMAP, 0.3 equiv., commercially available from MilliporeSigma, St. Louis, MO), 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide (EDCI, 3 equiv., MilliporeSigma) and dried CH.sub.2Cl.sub.2 (0.1 M). The resultant mixture was stirred for 18 hours, diluted with CH.sub.2Cl.sub.2, and then washed with water. The organic layer was collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which was purified by flash column chromatography to afford an ester compound as a colorless oil used in the next step directly as an intermediate.

[0178] At room temperature, the above intermediate (1 equiv.) was mixed with TFA (2.5 equiv.) and CH.sub.2Cl.sub.2 (0.2 M). The resultant mixture was stirred at room temperature for 16 hours and then concentrated under vacuo. The residue was purified by flash column chromatography to afford a carboxylic compound.

[0179] At room temperature, the above carboxylic compound (1 equiv.) was mixed with HATU (1.2 equiv.), DIPEA (4 equiv.) and compound S2 (1 equiv.). The reaction mixture was stirred at room temperature for 18 hours, diluted with CH.sub.2Cl.sub.2, and then washed with water. The organic layer was collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which was purified by prep HPLC (C18) to afford a compound 69: MS-ESI (m/z) calcd for C.sub.53H.sub.67N.sub.4O.sub.18 [M+H].sup.+ 1047.4450, found 1047.4.

[0180] Compounds 70, 71, and 72 were prepared following the method shown in Scheme XVII below.

##STR00108##

[0181] At room temperature, Compound SP001, Compound 7, or Compound 42 (1 equiv.) was mixed with succinic anhydride (3 equiv.), dimethylamino-pyridine (DMAP, 1.3 equiv., MilliporeSigma), DIPEA (3 equiv.) and dried CH.sub.2Cl.sub.2 (0.1 M). The resultant mixture was stirred at 40? C. for 18 hours, diluted with CH.sub.2Cl.sub.2, and then acidified with 1N HCl (aq.). The aqueous layer was extracted with CH.sub.2Cl.sub.2 for three times. The organic layers were collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which was purified by flash column chromatography to afford a carboxylic intermediate as a colorless oil.

[0182] At room temperature, the above intermediate (1 equiv.) was mixed with HATU (1.2 equiv.), DIPEA (4 equiv.) and compound S2 (1 equiv.). The reaction mixture was stirred at room temperature for 18 hours, diluted with CH.sub.2Cl.sub.2, and then washed with water. The organic layer was collected and dried over anhydrous MgSO.sub.4, filtered, and concentrated to obtain a residue, which is purified by prep HPLC (C18) to afford one of Compounds 70, 71 and 72.

[0183] Compound 70: MS-ESI (m/z) calcd for C.sub.53H.sub.69N.sub.4O.sub.18 [M+H].sup.+ 1049.4607, found 1049.5.

[0184] Compound 71: MS-ESI (m/z) calcd for C.sub.53H.sub.67N.sub.4O.sub.19 [M+H] 1063.4400, found 1063.4.

[0185] Compound 72: MS-ESI (m/z) calcd for C.sub.56H.sub.70N.sub.5O.sub.19 [M+H] 1116.4665, found 1117.5.

[0186] The compounds thus prepared can be further purified following conventional methods such as crystallization, distillation/vacuum distillation, flash chromatography over silica, and preparative liquid chromatography.

[0187] Efficacy of the compounds of this invention can be initially determined using an in vitro method to identify their anti-inflammatory activity, all described in examples below. The selected compounds can be further tested to verify their in vivo efficacy, pharmacokinetic profiles, and toxicity, e.g., by administering it to an animal. Based on the results, an appropriate dosage range and administration route can be determined.

[0188] A compound of this invention is preferably formulated into a pharmaceutical composition containing a pharmaceutical carrier. The pharmaceutical composition is then given to a subject in need thereof to inhibit inflammation thus treating disorders associated therewith.

[0189] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present invention to its fullest extent. The following examples are to be construed as merely illustrative and not limitative of the remainder of the disclosure in any way whatsoever.

[0190] All publications cited herein are hereby incorporated by reference in their entirety.

[0191] Set forth below are examples illustrating preparation and efficacy evaluation of compounds of this invention.

Real-Time PCR Analysis for iNOS mRNA

[0192] Real time quantitative polymerase chain reaction (qPCR) was performed according to methods described by Livak et al. and De Gois et al. with modifications. See Livak et al., Methods 2001, 25, 402-408; and De Gois et al., J. Neurosci. 2005, 25, 7121-33.

[0193] Cell pellets were collected from centrifuged tubes after 8 hours. Total RNA was isolated using TRIzol? RNA Isolation Reagents (Life Technologies, Carlsbad, California) according to the manufacturer's instructions. After centrifugation at 3000 rpm for 8 minutes at 4? C., total RNA was obtained and transcribed using the iScript? cDNA synthesis kit (Bio-Rad, Hercules, California). Reactions were performed in duplicate with 0.5 ?L of each primer (0.2 ?M final concentration), 25 ?L of iQ SYBR Green Supermix? (Bio-Rad, 100 mM KCl, 40 mM Tris-HCl, pH 8.4, 0.4 mM of each dNTP, iTaq DNA polymerase, 50 units/mL, 6 mM MgCl2, SYBR Green? I, 20 nM fluorescein and stabilizer) and 2.5 ?L of template in a 50-?L total volume. The PCR cycle conditions were 95? C. for 10 minutes, 40 cycle of 95? C. for 15 seconds, and then 60? C. for 1 minute. A melting curve analysis was performed at the end of each experiment to verify that a single product per primer pair was amplified. The amplification and analysis were performed using a CFX96 Touch? Real-time PCR Detection System (Bio-Rad). Results were compared using the relative cycle threshold (CT) method. The fold increase or decrease was determined relative to a blank control after normalizing to a housekeeping gene (GAPDH) using 2-??CT. The real-time PCR oligonucleotide primers used for genotyping are as follows: iNOS (forward), 5-GCTGTTAGAGACACTTCTGAG-3; iNOS (reverse), 5-CACTTTGGTAGGATTTGACTTTG-3.

[0194] The inhibition of iNOS (%) was calculated as percent reduction of iNOS gene expression to that of LPS-stimulated iNOS gene expression as 100%. A high iNOS inhibition indicates that the compound can exert a reduction of LPS-stimulated iNOS gene expression greater than 80%.

[0195] Compounds 1-67 were evaluated by the assay described above. Each compound showed inhibition of iNOS gene expression up to 97%. Among them, Compounds 24, 34, 37, 41, 55, and 66 were very potent, having an inhibition of iNOS gene expression between 50% and 97%.

OTHER EMBODIMENTS

[0196] All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0197] From the above description, one skilled in the art can easily ascertain the essential characteristics of the present invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. For example, compounds structurally analogous to the compounds of this invention also can be made, screened for their efficacy in treating a condition that relates to SOS1. Thus, other embodiments are also within the claims.